ARTICLE | doi:10.20944/preprints202210.0012.v1
Subject: Biology, Animal Sciences & Zoology Keywords: animal welfare; welfare range; comparative cognition; interspecies comparisons
Online: 3 October 2022 (13:03:23 CEST)
The number of animals bred, raised, and slaughtered each year is on the rise, resulting in increasing impacts to welfare. Farmed animals are also becoming more diverse, ranging from pigs to bees. The diversity and number of species farmed invites questions about how best to allocate currently limited resources towards safeguarding and improving welfare. This is of the utmost concern to animal welfare funders and effective altruism advocates, who are responsible for targeting the areas most likely to cause harm. For example, is tail docking worse for pigs than beak trimming is for chickens in terms of their pain, suffering, and general experience? Or are the welfare impacts equal? Answering these questions requires making an interspecies welfare comparison; a judgment about how good or bad different species fare relative to one another. Here, we outline and discuss an empirically-based methodology that aims to improve our ability to make interspecies welfare comparisons by investigating welfare range, which refers to how good or bad animals can fare. We begin our proposal with a theory of welfare. We operationalize that theory of welfare by identifying metrics that are defensible proxies for measuring welfare, including cognitive, affective, behavioral, and neuro-biological measures. We assign differential weights to those proxies that reflect their evidential value for the determinants of welfare, such as the “Delphi'' structured deliberation method with a panel of experts. Then we review the evidence and score its quality to ascertain whether a particular taxa may possess the proxies in question to construct a taxa-level welfare range profile. Finally, we use a Monte Carlo simulation to generate an overall estimate of comparative welfare range relative to our hypothetical index species - humans. Interspecies welfare comparisons will help facilitate empirically informed decision-making to streamline the allocation of resources and to ultimately better prioritize and improve animal welfare.
REVIEW | doi:10.20944/preprints202209.0473.v1
Subject: Biology, Animal Sciences & Zoology Keywords: seminal fluid; seminal plasma; epigenetics; transgenerational plasticity; sperm; paternal effects
Online: 30 September 2022 (03:25:29 CEST)
The evidence supports the occurrence of environmentally induced paternal epigenetic inheritance shaping the offspring phenotype in the absence of direct or indirect paternal care, and the empirical results clearly indicate that sperm epigenetics is one of the major actors mediating these paternal effects. However, sperm often make up only a small fraction of the male ejaculate in animals. Males also have a complex mixture of proteins, peptides, types of small RNAs, and cell-free DNA fragments in their seminal fluid. These molecules are in close contact with reproductive cells, tissues, organs, and other molecules of both males and females during reproduction. Moreover, their production and use are very sensitive to environmental conditions which makes them potential modulators of environmentally and developmentally induced paternal effects on the next generation(s). Although there is some intriguing evidence of seminal fluid-mediated paternal epigenetic effects, the underlying molecular mechanisms remain poorly defined. In this review, I discuss the current evidence regarding the association between seminal fluid and environmentally induced paternal effects, the possible trajectories, and the mechanisms in which seminal fluid can involve to mediate paternal epigenetic inheritance.
ARTICLE | doi:10.20944/preprints202208.0442.v2
Subject: Biology, Animal Sciences & Zoology Keywords: bison; restoration; socio-ecological processes; indigenous harvest; maximum entropy modelling
Online: 29 September 2022 (10:58:10 CEST)
The historic western edge of bison (Bison bison) range and the ecological processes that caused its formation are frequently debated with important implications for bison restoration across North America. We test the hypothesis that a combination of bottom-up habitat suitability and top-down harvest pressure from humans were important processes in forming the western edge of bison distribution. Using 9,384 historical journal observations from 1691 – 1928, we employ MaxEnt ecological niche modelling to identify suitable bison habitat across the Western Cordillera from bottom-up climatic, land cover, and topographic factors. We then use mixed-effect logistic regression to test if bison occurrence in journal records can be in part explained by the abundance of Indigenous humans, wolves, or grizzly bears, in addition to MaxEnt-derived habitat suitability. We find support for our hypothesis because of the limited suitable habitat in the Rocky Mountains that likely prevented westward bison dispersal from core habitat, and there was a negative relationship between bison occurrence and human harvest pressure. On this basis, we propose that intensive human harvest from large populations in the Western Cordillera, subsidized by other wildlife, salmon, and vegetation resources, is an underappreciated socioecological process that needs to be restored alongside bison populations. Co-managing bison with Indigenous people will also mitigate the adverse effects of overabundant bison and maximize the ecological and cultural benefits of bison restoration.
ARTICLE | doi:10.20944/preprints202209.0418.v1
Subject: Biology, Animal Sciences & Zoology Keywords: circRNA; skeletal muscle satellite cells; proliferation; differentiation
Online: 27 September 2022 (09:57:42 CEST)
The proliferation and differentiation of mammalian skeletal muscle satellite cells (MuSCs) are highly complicated. Apart from the regulatory signaling cascade driven by the protein-coding genes, non-coding RNAs like microRNAs (miRNA) and circular RNAs (circRNAs) play essential roles in this biological process. However, circRNA functions in MuSCs proliferation and differentiation remain largely to be elucidated. Here, we screened for an exonic circTCF4 based on our previous RNA-Seq data, specifically expressed during the development of the longest dorsal muscle in goats. Subsequently, the circular structure and whole sequence of circTCF4 were verified using Sanger sequencing. Besides, circTCF4 was spatiotemporally expressed in multiple tissues from goats but strikingly enriched in muscles. Furthermore, circTCF4 suppressed MuSCs proliferation and differentiation, independent of AGO2 binding. Finally, we conducted Poly(A) RNA-Seq using cells treated with small interfering RNA targeting circTCF4 and found that circTCF4 would affect multiple signaling pathways, including insulin signaling pathway and AMPK signaling pathway related to muscle differentiation. Our results provide additional solid evidence for circRNA regulating skeletal muscle formation.
REVIEW | doi:10.20944/preprints202209.0037.v1
Subject: Biology, Animal Sciences & Zoology Keywords: Multidrug-resistant Acinetobacter baumannii; Phage therapy
Online: 2 September 2022 (09:51:24 CEST)
Acinetobacter baumannii is a multidrug-resistant and invasive pathogen associated with the etiopathology of both an increasing number of nosocomial infections and of relevance to poultry production systems. Multidrug-resistant Acinetobacter baumannii has been reported in connection to severe challenges to clinical treatment, mostly due to an increase rate of resistance to carbapenems. Amid the possible strategies aiming to reduce the insurgence of antimicrobial resistance, phage therapy has gained particular importance for the treatment of bacterial infections. This review summarises the different phage-therapy approaches currently in use for multiple-drug resistant Acinetobacter baumannii, including single phage therapy, phage cocktails, phage -antibiotic combination therapy, phage coding Acinetobacter baumannii and the novel phage enzyme treatment. Although phage therapy represents a potential treatment solution for multidrug-resistant Acinetobacter baumannii, further research is needed to unravel some unanswered questions especially in regard to its in vivo applications, before possible routine clinical use.
REVIEW | doi:10.20944/preprints202208.0311.v1
Subject: Biology, Animal Sciences & Zoology Keywords: zoonotic pathogens; mathematical algorithms; machine learning; deep learning
Online: 17 August 2022 (08:57:27 CEST)
Globally, zoonotic diseases have been on the rise in recent years. Predictive modelling approaches have been successfully used in the literature to identify the underlying causes of these zoonotic diseases. We examine the latest research in the field of predictive modeling that verifies the growth of zoonotic pathogens and assesses the factors associated with their spread. The results of our survey indicate that popular mathematical models can successfully be used in modeling the growth rate of these pathogens under varying storage temperatures. Additionally, some of them are used for the assessment of the inactivation of these pathogens based on various conditions. Based on the results of our study, machine learning models and deep learning are commonly used to detect pathogens within food items and to predict the factors associated with the presence of the pathogens.
ARTICLE | doi:10.20944/preprints202208.0247.v1
Subject: Biology, Animal Sciences & Zoology Keywords: animal welfare; pain; farm animals; Pain-Track; Cumulative Pain; pain assessment; welfare foot-print; time; interspecific comparisons
Online: 15 August 2022 (03:57:59 CEST)
We describe a recently developed approach to quantify welfare loss in animals, the Cumulative Pain metric. It combines the two most relevant dimensions of negative affective experiences: intensity and duration. The metric enables estimating the time individuals spend in negative affective states of a physical or psychological nature (operationally referred to simply as ‘pain’) of different intensities as the result of one or more challenges (e.g., diseases, injuries, deprivations). A new notation protocol (the Pain-Track) is used in which the duration of the experience is represented along the horizontal axis and intensity is represented by four categories in the vertical axis. Pain experiences are partitioned into temporal segments, where hypotheses for the experienced duration and intensity are proposed based on existing welfare indicators (e.g., neurophysiological, behavioral, anatomical, evolutionary). This structure forces transparency about assumptions and uncertainties, highlights knowledge gaps, and enables estimates to be continuously adjusted. Because the Cumulative Pain metric is based on parameters with a broadly common biological meaning, it provides the much needed interoperability among assessments of animal welfare. It enables comparing the impact of practices and living conditions, policies and interventions, and the calculation of welfare footprints of animal-sourced products using a universal measurement unit.
ARTICLE | doi:10.20944/preprints202208.0139.v1
Subject: Biology, Animal Sciences & Zoology Keywords: blackfin reef shark; Carcharhinus melanopterus; shark behaviour; shark ethology; shark cognition
Online: 8 August 2022 (09:44:57 CEST)
The chondrichthyan lineage diverged from the osteichthyan line 440 million years ago, resulting in a vast evolutionary gulf between modern elasmobranchs and other vertebrates. Though this has supported the assumption that sharks are ancient, dangerous, and binary-minded, the few ethological studies done have noted intelligent actions including social exchanges. Yet their behaviour remains little known. On seeing that Carcharhinus melanopterus displayed complex actions during incidental meetings, a long-term ethological study of the species was carried out using artificial aggregations, at several sites in the fringe lagoon of Mo’orea Island, French Polynesia. Short and long-term behaviour was recorded in 473 individuals, including an ethogram, roaming patterns, social interactions, and cognition. C. melanopterus is considered sedentary, yet the home range could also be viewed as a place to pause between travels, for most individuals left for long periods. The study community and its visitors travelled in correlation with the lunar phase, in groups of up to six individuals, socializing with conspecifics encountered along the way, and displaying fluid social dynamics. C. melanopterus was highly alert to danger yet prone to investigate novel objects, a combination that generated a variety of tactics to remain hidden while investigating the environment. Basic to this was the use of the visual limit for escape or to screen their presence, indicating an awareness of being present and observable. Using their other senses, they could focus their attention on events beyond visual range and made swift decisions to act as circumstances unfolded. In their non-territorial, non-hierarchical society, any shark could lead, but it was usually the same ones that did so. Therefore, unusual individuals had a significant effect on events through social learning, suggesting the potential for culture. Actions in a variety of situations suggested complex cognition, and individuals displayed both positive and negative subjective states including playfulness.
ARTICLE | doi:10.20944/preprints202208.0057.v1
Subject: Biology, Animal Sciences & Zoology Keywords: infectious bronchitis; viral evolution; whole genome sequencing; DMV; QX.
Online: 2 August 2022 (09:27:23 CEST)
Infectious bronchitis virus (IBV) is a highly variable RNA virus that affects chickens worldwide. Due to its inherited tendency to suffer point mutations and recombination events during viral replication, emergent IBV strains have been linked to nephropathogenic and reproductive disease that are more severe than the typical respiratory disease, leading, in some cases, to mortality, severe production losses, and/or unsuccessful vaccination. QX and DMV/1639 strains are examples of the above-mentioned IBV evolutionary pathway and clinical outcome. In this study, our purpose was to systematically compare whole genomes of QX and DMV strains looking at each IBV gene individually. Phylogenetic analyses and amino acid site searches were performed in datasets obtained from GenBank accounting for all IBV genes and using our own relevant sequences as a basis. The QX dataset studied is more genetically diverse than the DMV dataset, partially due to the greater epidemiological diversity within the five QX strains used as a basis compared to the four DMV strains from our study. Historically, QX strains have emerged and spread earlier than DMV strains in Europe and Asia. Consequently, there are more QX sequences deposited in GenBank than DMV strains, assisting in the identification of a larger pool of QX strains. It is likely that a similar evolutionary pattern will be observed among DMV strains as they develop and spread in North America.
ARTICLE | doi:10.20944/preprints202207.0414.v1
Subject: Biology, Animal Sciences & Zoology Keywords: dogs; Colombia; leptospirosis; Leptospira santarosai; seroreactivity; risk factors
Online: 27 July 2022 (08:07:27 CEST)
Canine leptospirosis is a zoonosis of epidemiological importance. Dogs are recognized as primary reservoirs of Leptospira interrogans serogroup Canicola and a source of infection to the environment through urine. This study aimed to determine the presence of antibodies against Leptospira in canines from 49 municipalities in the Department of Antioquia, Colombia. We performed a cross-sectional study of dogs included in a neutering control program. We collected 1335 sera samples, assayed by a microagglutination test (MAT), and performed PCR detection in 21 urine samples. We also surveyed 903 dog owners. We found a seroreactivity of 11.2% (150/1335) in Antioquia with titers 1:50. Municipalities with the highest number of cases were Belmira (46.1%), Turbo (34.5%), and Concepción (31.0%). L. santarosai was identified by phylogenetic analysis in one urine sample from the municipality of Granada. The most important factor associated with a positive result was the lack of vaccination against leptospirosis (PR 3.3, p < 0.014). Environmental factors such as water presence and bare soil around the household were also associated with Leptospira seroreactivity in the Department of Antioquia. We reviewed a national epidemiological surveillance database for human cases in those municipalities. We found a correlation between the high number of cases in canines and humans, especially in the Uraba. Serological and molecular results showed the circulation of Leptospira. Future public health efforts in the municipalities with the highest numbers of seroreactivity should be directed towards vaccination to prevent animal disease and decrease the probability of transmission of Leptospira. Dogs actively participate in the Leptospira cycle in Antioquia and encourage the implementation of vaccination protocols and coverage.
REVIEW | doi:10.20944/preprints202207.0200.v1
Subject: Biology, Animal Sciences & Zoology Keywords: microRNAs; Precision livestock science; animal welfare; livestock health; biomarkers; biosensor; pandemics
Online: 13 July 2022 (13:12:32 CEST)
Early disease detection in livestock allows for target treatment decreasing antibiotics use and allow advancements in precision veterinary medicine. MicroRNA (miRNA) -driven signaling cascades play a crucial role in the context of farm animal disease diagnostics and prediction, and their proper understanding remains a challenge. In livestock farm animals, only a small number of miRNAs have been fully validated with respect to disease conditions and physiological or behavioral traits. Low abundance of miRNAs in blood and bodily fluids, along with a small number of nucleotides, makes detection and discrimination tedious and challenging task in. miRNAs usually are homologous, owing to which detection specificity becomes next to impossible when screening for multiple miRNAs in the same analyte sample. Hence, a concurrent, multiplexing, approach becomes crucial for the development of on-farm point-of-care based detection systems. Comprehensive screening methods demand broad dynamic range and enhanced specificity. For on-farm handheld platform development, the ability to screen for multiple varieties of miRNA is essential. In this review paper, I provide an overview of the recent developments of miRNA sensing and the current bottlenecks in the realization of the sensors for detecting miRNAS as target analyte for various livestock disease detection applications. Due to the nascent stages of this research, the possibilities of exploiting miRNAs as a biomarker opens up ways to move from reactive to predictive possibilities in diseases detection in the modern digital livestock farming.
TECHNICAL NOTE | doi:10.20944/preprints202207.0109.v1
Subject: Biology, Animal Sciences & Zoology Keywords: Heterochromatin; mealy bugs; parent-of-origin effects; genomic imprinting
Online: 7 July 2022 (04:22:14 CEST)
Study of imprinted heterochromatinization of the paternal chromosome set in male mealy bugs is made difficult because it takes place at the blastula stage within the ovary. We describe here a method that allows for the bulk preparation of staged early embryos that develop normally outside the mother. We define an accessible experimental window encompassing 48 to 72hours post-mating in which regulation of heterochromatinization of the paternal chromosome set can now be investigated outside the confines of the ovary.
ARTICLE | doi:10.20944/preprints202207.0063.v1
Subject: Biology, Animal Sciences & Zoology Keywords: Native shrubs; In vitro fermentation; volatile fatty acids; greenhouse gases; hill country
Online: 5 July 2022 (07:40:50 CEST)
Information on the nutritive value and in vitro fermentation characteristics of native shrubs in New Zealand is scant. This is despite their potential as alternatives to exotic trees and shrubs for sup-plementary fodder, and mitigation of greenhouse gas and soil erosion on hill country sheep and beef farms. The objectives of this study were to measure the in vitro fermentation gas production, predict parameters of in vitro fermentation kinetics and to estimate in vitro fermentation of volatile fatty acids (VFA), microbial biomass (MBM) and greenhouse gases of four native shrubs (Coprosma robusta, Griselinia littoralis, Hoheria populnea and Pittosporum crassifolium) and an exotic fodder tree species, Salix schwerinii. Total in vitro gas production was higher (p<0.05) for natives than S. schwerinii. Prediction using the single pool model resulted in biologically incorrect negative in vitro total gas production from the immediately soluble fraction of the native shrubs. However, the dual pool model better predicted in vitro total gas production and was in alignment with measured in vitro fermentation end products. In vitro VFA and greenhouse gas production from fermentation of leaf and stem material were higher (p<0.05), and MBM lower (p<0.05), for native shrubs com-pared to S. schwerinii. The lower in vitro total gas production, VFA and greenhouse gases produc-tion, and higher MBM of S. schwerinii may be explained by the presence of condensed tannins (CT), although this was not measured and requires further study. In conclusion, results from this study suggests that when consumed by ruminant livestock, the browsable native shrubs can provide adequate energy and microbial protein, and that greenhouse gas production from these species is within ranges reported for typical New Zealand pastures.
ARTICLE | doi:10.20944/preprints202206.0430.v2
Online: 4 July 2022 (10:57:51 CEST)
Humans, willingly or unwillingly, affect the lives of urban cats. Waste production is only one of the human effects on the urban cat ecosystem. The human-generated food waste provides a large portion of urban cat's food, and economic fluctuations change the quality, and content of waste bins. In this descriptive-estimation study with field monitoring, we tried to measure the weight of cats to find out how much the change in the volume of municipal food waste affects their physical condition? The results of a study conducted in Tehran from spring 2016 to winter 2020 showed that the average volume of food waste collected in this city had decreased by 26.9%. The reduction of food waste volume affected the weight of cats and caused an average of 18.71% reduction in the average weight of cats during the study. The weight loss rate intensified from autumn 2017 – to winter 2018 onwards. Male cats lost 18.68%, and female cats 18.40% of their weight from early spring 2016 to late winter 2020.
ARTICLE | doi:10.20944/preprints202202.0211.v2
Subject: Biology, Animal Sciences & Zoology Keywords: pIgR; gene structure; cold environment; gene expression; teleost immunity; adaptive evolution; mucosal tissues; genome alteration; Notothenioidei; IgV domains.
Online: 27 June 2022 (08:41:00 CEST)
The IgM and IgT classes were previously identified and characterized in the Antarctic teleost Trematomus bernacchii, a species belonging to the Perciform suborder Notothenoidei. Herein we characterized the gene encoding the polymeric immunoglobulin receptor (pIgR) in the same species and compared it to pIgR of multiple teleost species belonging to five perciform sub-orders, including 11 Antarctic and one non-Antarctic (Cottoperca gobio) notothenioid species, the latter living in less cold periantarctic sea. Antarctic pIgR genes displayed particularly long in-trons marked by sites of transposable elements and transcription factors. Furthermore, analysis of T. bernacchii pIgR cDNA unveiled multiple amino acid substitutions unique to Antarctic species, all introducing adaptive features, including N-glycosylation sequons. Interestingly, C. gobio shared most features with the other perciforms rather than with the cold adapted relatives. T. bernacchii pIgR transcripts were predominantly expressed in mucosal tissues, as indicated by q-PCR and in situ hybridization analysis. These results suggest that in cold adapted species pIgR preserved its fundamental role in mucosal immune defense, although remarkable gene structure modifications occurred.
ARTICLE | doi:10.20944/preprints202206.0268.v1
Online: 20 June 2022 (10:10:54 CEST)
Although sharks have a fundamental role in maintaining the balance of aquatic ecosystems, exerting a great influence at lower levels, their populations are declining worldwide due, to a large extent, to overfishing. Of the 64 species registered in Ecuador, from January to December 2019, 19 species were recorded in Manta from 15455 captured individuals, with the family Carcharhinidae being the family most present in the catches (69.4%), and the most abundant species was Prionace glauca (57.9%). In the case of threatened species, such as Carcharhinus longimanus, Sphyrna lewini and Sphyrna zygaena, a greater presence of immature specimens was observed in landings, suggesting a possible existence of nursery areas. However, information on the composition and biological aspects of shark species in the Ecuadorian Pacific is very scarce. Therefore, research on the characteristics of life history (age, growth and maturity) are of utmost importance for the analysis evolution of the populations that are being exploited, especially in developing countries, where this information is very scarce, causing inadequate management of fishery resources.
ARTICLE | doi:10.20944/preprints202206.0207.v1
Subject: Biology, Animal Sciences & Zoology Keywords: Germinal Center; Herpesvirus; Recombinant protein; rgD5; Vaccine; Long-Lasting
Online: 14 June 2022 (16:21:47 CEST)
Bovine herpesvirus (BoHV)-5 is a worldwide distributed pathogen usually associated with a lethal neurological disease (meningoencephalitis) in dairy and beef cattle resulting in important economic losses due to the cattle industry. Using recombinant glycoprotein D of BoHV-5 (rgD5), we evaluated the long-duration humoral immunity of the recombinant vaccines in a cattle model. Here we report that two doses of intramuscular immunization, particularly with the rgD5ISA vaccine, are superior to iBoHV-5ISA immunization in the induction of long-lasting antibody responses. Recombinant gD5 antigen elicited tightly mRNA transcription of the Bcl6 and the chemokine receptor CXCR5 which mediate memory B cells and long-lived plasma cells in germinal centers (GCs). In addition, using an in-house Enzyme-Linked Immunosorbent Assay (ELISA) we observed higher and earlier responses of rgD5-specific IgG antibody and the upregulation of mRNA transcription of IL2, IL4, IL10, IL15 and IFN-γ cytokines in rgD5 vaccinated cattle, indicating a mixed immune response. We further show that rgD5 immunization provides protection against both BoHV -1 and -5. Our findings indicate that the rgD5-based vaccine represents an effective vaccine strategy to induce an efficient control of alpha-herpesviruses.
ARTICLE | doi:10.20944/preprints202206.0178.v1
Subject: Biology, Animal Sciences & Zoology Keywords: Citizen science; Colour pattern; geographic diversity; phylogeography
Online: 13 June 2022 (09:55:52 CEST)
The geographic variability of the dorsal pattern (DP) of the Italian wall lizard, Podarcis siculus, across its native range was studied with the aim to understand whether the distributions of this phenotypic trait were more shaped by allopatric differentiation rather than adaptive processes. A total of 1298 georeferenced observations scattered across the Italian peninsula and the main islands (Sicily, Corsica and Sardinia) were obtained from citizen science databases and five DPs were characterized by different shapes of the dark pattern (“reticulated”, “campestris”, “reticulated/campestris” and “striped”) or by absence of it (“concolor”). Frequencies of different DP phenotypes differ between the two main mtDNA lineages settled in central-northern and in southern Italy respectively. This pattern may be indicative of a role of long-term allopatric historical processes in determining the observed pattern. The analysis also identified a putative wide area of secondary contact, in central southern Italy, characterized by high diversity of the DP. Generalized Linear Models (GLMs), used to estimate a possible association between bioclimatic variables and the observed phenotypic variation, showed that each of the five DPs is correlated to different environmental factors and show different distribution of areas with high probability of occurrence. However, for all but one of the DPs, the area with the greatest probability does not correspond exactly to the real distribution of the DP. Conversely, the “concolor” phenotype does not seem related to any particular mtDNA lineage and it shows a preference for areas with high temperature and low rainfall. This is in agreement with the expectation of low amount of melanin of the dorsal pattern that, in the study areas, is characterized by a light uniform coloration which could confer a better thermoregulation ability in high temperatures environments avoiding overheating.
ARTICLE | doi:10.20944/preprints202206.0125.v1
Subject: Biology, Animal Sciences & Zoology Keywords: PCSK7; miR-125a-5p; miR-143-3p; miR-409-3p; RNA regulator; proprotein convertases
Online: 8 June 2022 (10:45:01 CEST)
The regulatory mechanism of PCSK7 gene is still unknown, although its encoded protein PC7 is the most ancient and highly conserved of all proprotein convertases and exhibits enzymatic and non-enzymatic functions in liver triglyceride regulation. Bioinformatics algorithms were used to predict regulatory microRNAs (miRNAs) of PCSK7 expression. This led to the identification of 4 miRNAs, namely miR-125a-5p, miR-143-3p, miR-409-3p, and miR-320a-3p, with potential binding sites on the 3’-untranslated region (3′-UTR) of human PCSK7 mRNA. The expression patterns of these miRNAs and PCSK7 mRNA were assessed in three different cell lines with quantitative polymerase chain reaction (qPCR), which revealed reciprocal expression patterns between the expression levels of the four selected miRNAs and PCSK7. Next, the interactions and effects of these miRNAs on PCSK7 expression levels were investigated via cell-based expression analysis, dual-luciferase assay, and Western blot analysis. The data revealed that PCSK7 mRNA levels decreased in cells transfected with vectors overexpressing miR-125a-5p, miR-143-3p, and miR-409-3p, but not miR-320a-3p. The dual-luciferase assay demonstrated that the above 3 miRNAs could directly interact with putative target sites in PCSK7 3′-UTR and regulate its expression, whereas miR-320-3p exhibited no interaction. Western blot analysis further revealed that overexpression of miR-125a-5p in Huh7 cells inhibits the expression and ability of PC7 to cleave human transferrin receptor 1. Our results support a regulatory role of these miRNAs in PCSK7 expression and function and opens the way to assess their roles in the regulation of PC7 activity in vivo in the development of hepatic steatosis.
COMMUNICATION | doi:10.20944/preprints202206.0015.v1
Subject: Biology, Animal Sciences & Zoology Keywords: barcoding; Dactylogyridae; Dactylogyrinae; ectoparasites; giant goby; Gobiidae; Gobius cobitis; Greece; Monogenea; Platyhelminthes
Online: 1 June 2022 (10:46:18 CEST)
Gobies and their ectoparasitic monogenean flatworms are promising models for species diversification because of their species richness. Recent decades have seen the discovery of several new species of Gyrodactylus (Monogenea: Gyrodactylidae) on European gobies, mostly in the sand goby lineage and especially in the Eastern Mediterranean. However, the monogenean fauna of other gobies is much less understood. Therefore, we scrutinized five gobiid species (34 specimens, vouchered, with some representatives sequenced), sampled in Greece, for monogenean ectoparasites. Only specimens of the giant goby Gobius cobitis were infected; they harbored Xenoligophoroides cobitis (Monogenea: Dactylogyridae) on their gills. Here we provide the first record from Greece, and the first ITS rDNA and COI sequences of the representative of this monotypic genus. Also 28S rDNA was sequenced and compared with published data from across its known distribution, suggesting clinal variation. Lack of sequence data of closely related dactylogyrid monogeneans prevents either proposing a sister-group for Xenoligophoroides, or inferring a scenario explaining the presence of a single known member of this genus on gobies. Possible hypotheses include either the ancestral long-term presence on gobiids but “missing the boat” of the diversification events in the “Gobius-lineage”, or a recent host switch from a non-gobiid host.
CASE REPORT | doi:10.20944/preprints202205.0397.v1
Online: 30 May 2022 (11:27:09 CEST)
Tawny nurse shark (Nebrius ferrugineus) is a species of carpet shark distributed across the tropical Indo-Pacific region. Tawny nurse shark is listed as “Vulnerable” on IUCN Redlist and rarely seen, recorded and studied in Indonesia. We recorded this species during SCUBA diving activity in Morotai waters in April 2022. Scientific record of the species is highly important for conservation and habitat management, particularly in Morotai, a region in North Maluku famous for shark-based marine tourism.
BRIEF REPORT | doi:10.20944/preprints202205.0324.v1
Subject: Biology, Animal Sciences & Zoology Keywords: aortic aneurysms; aortic dissection; fludrocortisone; angiotensin; hypercholesterolemia; mouse
Online: 24 May 2022 (06:05:35 CEST)
Background and Objective: In an experiment designed to explore the mechanisms of fludrocortisone-induced high blood pressure, we serendipitously observed aortic aneurysms in mice infused with fludrocortisone. The purpose of this study was to investigate whether fludrocortisone induces aortic pathologies in both normocholesterolemic and hypercholesterolemic mice. Methods and Results: Male adult C57BL/6J mice were infused with either vehicle (85% polyethylene glycol 400 (PEG-400) and 15% dimethyl sulfoxide (DMSO); N=5) or fludrocortisone (12 mg/kg/day dissolved in 85% PEG-400 and 15% DMSO; N=15) for 28 days. Fludrocortisone-infused mice had higher systolic blood pressure, compared to mice infused with vehicle. Fludrocortisone induced aortic pathologies in 4 of 15 mice with 3 having pathologies in the ascending and aortic arch regions and 1 having pathology in both the ascending and descending thoracic aorta. No pathologies were noted in abdominal aortas. Subsequently, we infused either vehicle (N=5/group) or fludrocortisone (N=15/group) into male ApoE -/- mice fed a normal laboratory diet or LDL receptor -/- mice fed either normal or Western diet. Fludrocortisone increased systolic blood pressure, irrespective of mouse strain or diet. In ApoE -/- mice infused with fludrocortisone, 2 of 15 mice had ascending aortic pathologies, but no mice had abdominal aortic pathologies. In LDL receptor -/- mice fed normal diet, 5 had ascending/arch pathologies, 1 had pathologies in the ascending, arch, and suprarenal aortic regions. In LDL receptor -/- mice fed Western diet, 2 died of aortic rupture in either the descending thoracic or abdominal region, and 2 of the 13 survived mice had ascending/arch aortic pathologies. Aortic pathologies included hemorrhage, wall thickening or thinning, or dilation. Given the low incidence, only ascending aortic diameter in LDLR -/- mice fed Western diet reached statistical significance, compared to their vehicle. Conclusion: Fludrocortisone induces aortic pathologies independent of hypercholesterolemia. The findings in mouse studies have the potential to provide caution to people who are taking or have taken fludrocortisone that could have an increased risk of aortic pathologies.
ARTICLE | doi:10.20944/preprints202205.0221.v1
Subject: Biology, Animal Sciences & Zoology Keywords: ring necked-pheasant; ascorbic acid; semen cryopreservation; semen quality
Online: 17 May 2022 (05:19:30 CEST)
Ring necked-pheasant (Phasianus colchicus) is bird of order Galliformes. Ring necked-pheasant is also known as gallinaceous bird and game bird. It looks like chicken bird. It is national bird of South Dakota. It is mostly found in wild areas but in Pakistan it exists as Domestic bird. It has high proteins and low fat in its meat. The present study was conducted to check the effect of ascorbic acid on sperm Motility, sperm cytoplasmic membrane integrity, sperm livability and acrosome integrity of Ring-necked pheasant at different concentrations of ascorbic acid (0mM, 1mM, 2mM, 3mM and 4mM) were used. Semen was collected by abdominal massage technique. Motility of sperm was greater than 70%.Then it was processed further. The cryopreservation of semen was checked at various stages like Post Dilution, Post Cooling, Post Equilibration and Post Thawing. Semen was cooled from post-dilution stage (370 C) to post-cooling (200C).Then it further cooled gradually to get post-equilibration stage (40 C) within 24 hours. After that 10% glycerol was added to the sample. Then it was transferred to liquid nitrogen (LN2) cylinder for 24 hours. After thawing stage sample was removed from LN2 cylinders and placed in water bath for 4 hours to achieve post-thawing. Then performed sperm quality assays at each stage of cryopreservation. Sperm motility was assessed by Neubauer Chamber Hemocytometer. Plasma Membrane Integrity was checked by using Hypo Osmotic Swelling Test with the help to 2% Eosin. Sperm livability was assessed by Lake’s glutamate solution and sperm Acrosome Integrity was checked by dual staining technique by using Formal citrate solution and Giemsa stain. The better results were seen at 3mM treatment of ascorbic acid. Sperm motility percentage was significantly different (P < 0.05) on 3mM treatment on all the stages of cryopreservation rather than other treatments. Plasma membrane integrity assay also showed good results at same concentration (P < 0.05) on all the stages of cryopreservation rather than other treatments. At the sperm livability stage 3mM treatment showed significantly difference (P < 0.05) on all the stages of cryopreservation rather than other treatments. The sperm acrosome integrity showed highest percentage on 3mM treatment (P < 0.05) on all stages of cryopreservation rather than other treatments. Effect of ascorbic acid on all the quality parameters (sperm motility, plasma membrane integrity, livability and acrosome integrity) was showed significant difference (P < 0.05) at 3mM treatment of ascorbic acid as compare to control as well as remaining treatments. It was seen that by increasing the concentration of ascorbic acid from 3mM concentration to onward it shows negative results on its cryopreservation. Now this cryopreserved semen can be transfer from one place to another place to obtain good varieties as well as better genetic characteristics of this species.
ARTICLE | doi:10.20944/preprints202204.0195.v1
Subject: Biology, Animal Sciences & Zoology Keywords: crabs; fiddlers; sesarmids; burrowing; bioturbation; mangrove restoration
Online: 21 April 2022 (08:18:59 CEST)
Burrowing crabs are considered to be the ecosystem engineers in mangrove ecosystems because they have impact on the ecosystem functioning through bio-geochemical transformations. This process depends on the size and shape of burrows. The present study analysed architecture of burrows, constructed by crabs in a restored mangrove habitat. Fourteen crab species were found to construct the burrows of 13 different shapes with predominance of ‘I’,’J’,’L’. The burrow shape was diverse in Metopograpsus messor, followed by Austruca occidentalis, Gelasimus vocans, and A. annulipes. The burrow structural complexity was higher during summer in Avicennia or open zone. Sesarmids were larger in size than fiddlers, making the burrows of wider opening. Fiddlers were relatively smaller in size and they constructed complex burrows at vertical position, making longer and deeper burrows, in contrast to sesarmids, which formed simple burrows at horizontal position, digging shorter and shallower burrows. The sesarmids have smaller burrows without branching in mangrove zones, whereas the fiddler crabs (except Austruca variegata) have larger burrows with or without branching in open and Avicennia zones. The fiddler crabs especially Austruca occidentalis and A.annulipes have separate openings and passage for exit and entry as an adaptation against predators. The present work identified Austruca occidentalis and A. annulipes as the most potent bio-turbating crab species for restored mangrove habitats due to their efficiency in soil excavation and large sized burrows.
ARTICLE | doi:10.20944/preprints202204.0137.v1
Subject: Biology, Animal Sciences & Zoology Keywords: Taihe silky fowl; metabolic components; un-targeted metabolome; breed and feed; biosynthesis of amino acids
Online: 15 April 2022 (05:47:06 CEST)
Chinese Taihe Black-bone silky fowl (TBsf) is the homology of medicine and food and has high nutritional and medical value all over the world. However, the nutritional compositions and specific metabolite advantages of Taihe silky fowl muscle are still poorly understood. In this study, we investigated the differences of nutritional components between TBsf and another similar breed (Black Feathered chicken and laid green-shelled eggs, BF-gsc). Meanwhile, we also explored the divergences in muscle characteristics of Taihe silky fowl fed with two different diets, that is normal chicken feed (TBsf-ncf) and Broussonetia papyrifera-fermented feed (TBsf-bpf). Firstly, the growth performance and biochemical index of Taihe silky fowl was significantly different compared with black-feathered chicken. Secondly, we identified the metabolic alterations in Taihe silky fowl by performing an un-targeted UHPLC-Q-TOF-MS/MS analysis. Our results suggested that the whole metabonomic characteristics had obvious separation between TBsf-ncf, TBsf-bpf and BF-gsc groups both in the positive and negative ion mode by PCA analysis. Next, OPLS-DA multivariate analysis revealed that 57 metabolites (in positive mode) and 49 metabolites (in negative mode) were identified as differential metabolites between TBsf-ncf and BF-gsc group. These differential metabolites were mainly enriched to ABC transporters, biosynthesis of amino acids and aminoacyl-tRNA biosynthesis. Besides, there were 47 metabolites (in positive) and 13 metabolites (in negative) were differentially regulated between TBsf-ncf and TBsf-bpf group, which were majorly involved in histidine metabolism and linoleic metabolism. Furthermore, the integrated network analysis suggested that DL-arginine, DL-isoleucine, linoleoylcarnitine, stearoylcarnitine (positive) and ricionleic acid, D-proline, uric acid (negative) were the significantly metabolic biomarkers in Taihe silky fowl. Moreover, the metabolites of primaquine, ticlpoidine, riboflavin, acetylcarnitine (positive) and salicylic acid, acetaminophen sulfate, glutamic acid (negative) were markedly changed in the Taihe silky fowl fed with BP-fermented feed. In summary, a global survey of the nutritional components and metabolite differences were performed in muscle tissues of Taihe silky fowl between various breeds and feeds. The comprehensive expression profiles of the metabolites in Taihe silky fowl affected by genetic and environmental factors were acquired. This study provided valuable evidence fo breed and feed-induced putative biomarkers as well as improved the economic value of Taihe silky fowl through targeted metabolite regulation.
ARTICLE | doi:10.20944/preprints202203.0412.v1
Subject: Biology, Animal Sciences & Zoology Keywords: pipefish; Syngnathus acus; biology; ecology; reproduction; trophic plasticity; stable isotopes; diet
Online: 31 March 2022 (14:37:45 CEST)
The great pipefish Syngnathus acus is one of the most representative European syngnathid, being highly associated with seagrass and macroalgal beds. Surprisingly, the ecology of this ovoviviparous marine fish has received scanty attention. The population inhabiting three sites on Cíes Archipelago (Atlantic Islands National Park, NW Spain) was monitored in 2017-2018 for spatial and temporal changes in abundances, reproduction traits, trophic niche occupancy and dietary regimes across reproduction states through an isotopic (δ13C and δ15N) approach. Abundances were highly variable across seasons and sites, decreasing significantly from mid-autumn. The population consisted almost exclusively of large adults that migrate by the end of the breeding season, which extended from mid-spring to summer. Operational sex ratios suggest that the species is sex-role reversed. S. acus is a secondary consumer (Trophic position= 3.36 ± 0.05) preferring amphipods but displaying annual and seasonal dietary plasticity. Mature fish were less selective than immatures (especially females) with a higher preference for amphipods (36-68%) in the former. The second most preferred prey were carideans, copepods or isopods, depending on the year and reproduction state. Overall, the wider trophic niches in females and immature specimens compared to males and mature fish would indicate a higher variability in both the use of prey resources and/or their origin. The present study highlights the trophic plasticity and unique features of S. acus population in Cíes Archipelago, especially regarding the outstanding size of the fish and the exceptional occurrence of breeders.
ARTICLE | doi:10.20944/preprints202203.0394.v1
Subject: Biology, Animal Sciences & Zoology Keywords: hippocampus; seahorse; diet switch; stable isotopes; turnover; discrimination factor
Online: 30 March 2022 (15:19:41 CEST)
The initial development of seahorse juveniles is characterized by low digestion capabilities. Stable isotope analysis is an effective tool in studies of trophic food webs and animal feeding patterns. The present study provides new insights for the understanding of growth and food assimilation in early developing seahorses following a laboratory diet switch. The study was performed in early life stages of the seahorse Hippocampus reidi by assessing the influence of diet shift on changes and turnovers in carbon (δ13C) and nitrogen (δ15N) stable isotope in juveniles. Newborn seahorses were fed for 60 days following two feeding schedules (A6 and A11) based initially on copepods Acartia tonsa and subsequently on Artemia nauplii (since days 6 and 11, respectively). After prey shift, we determined δ13C and δ15N turnover rates as functions of change in either body mass (fitting model G) and days of development (fitting model D), contributions of metabolism and growth to those turnover rates, and diet-tissue discrimination factors. Survival, final dry weight and final standard length for diet A11 were higher compared to diet A6. The shift from copepods to Artemia lead to fast initial enrichments in δ13C and δ15N. Afterwards, the enrichment was gradually reduced until reaching the isotopic equilibrium with diet. In most cases, both fitting models performed similarly. The isotopic analysis revealed that 100% of tissue turnover was attributed to growth in diet A11, whereas 19-25% was endorsed to metabolism in diet A6. Diet-tissue discrimination factors were estimated for the first time in seahorse juveniles, resulting in higher estimates for diet A11 (2.9 ± 0.7‰ for δ13C; 2.5 ± 0.2‰ for δ15N) than in diet A6 (1.8 ± 0.1‰ for δ13C; 1.9 ± 0.1‰ for δ15N). This study highlights the relevance of feeding on copepods and their effect on isotopic patterns and discrimination factors in seahorse juveniles after a dietary shift. Regarding the application of the results achieved to feeding schedules in the rearing of H. reidi, a long period of feeding on copepods during the first days of development is highly recommended.
REVIEW | doi:10.20944/preprints202203.0324.v1
Subject: Biology, Animal Sciences & Zoology Keywords: metazoans; adhesive organs; suction organs; functional morphology; adhesive proteins
Online: 24 March 2022 (07:20:55 CET)
To resist hydrodynamic forces, two main underwater attachment strategies have evolved multiple times in aquatic animals: glue-like “bioadhesive secretions” and pressure-driven “suction attachment”. In this review, we use a multi-level approach to highlight convergence in underwater attachment mechanisms across four different length-scales (organism, organ, microscopic and molecular). At the organism level, the ability to attach may serve a variety of functions, the most important being: (i) positional maintenance, (ii) locomotion, (iii) feeding, (iv) building, and (v) defense. Aquatic species that use bioadhesive secretions have been identified in 28 metazoan phyla out of the 34 currently described, while suction organs have a more restricted distribution and have been identified in five phyla. Although biological adhesives are highly diverse, it is possible to categorize them into four main types according to the time scale of operation: permanent, temporary, transitory, and instantaneous adhesion. At the organ level some common principles have evolved independently in different biological lineages: for example, animals with single-unit attachment organs can be distinguished from those with multi-unit organs. Fundamental design elements can also be recognized for both types of attachment mechanisms. Suction attachment systems comprise a circular or elliptical attachment disc, a sealing rim to prevent leakage and a mechanism to lower the internal pressure. Bioadhesive-producing organs, on the other hand, usually contain a glandular tissue associated with connective tissues or other types of load-bearing support structures and muscles that facilitate locomotion or mechanical detachment. At the microscopic level, similar designs and organizations appear once again to have emerged independently in different phylogenetic lineages. Independent of the taxon and type of adhesion, there are species in which the biosynthesis, packaging and release of adhesive secretions takes place at the level of a single type of secretory cell, whereas in others these secretions are produced by two or more secretory cell types. Duo-gland adhesive systems involved in temporary adhesion present an additional level of complexity as they also exhibit de-adhesive secretory cells. Yet, strikingly similar cellular organizations have been reported in highly disparate species. In the case of biological suction organs, regions of the organ that contact the substratum are highly textured with stiff microstructures. Although clearly non-homologous in different animals, these microstructures are thought to enhance friction on rough surfaces. At the molecular level, proteins are the main organic constituent of adhesive secretions in aquatic animals. We compared the global amino acid compositions of bioadhesives using principal component analysis to show that homologous adhesives from phylogenetically related species cluster together, and there is little overlap between taxonomic groups. However, several non-permanent adhesives are grouped together even though they belong to disparate phyla, indicating convergence in amino acid composition. We also investigated relatedness among individual adhesive proteins using a sequence similarity-based clustering analysis. While many proteins appear taxon-specific, some have clear sequence homologies based on shared protein domains between phylogenetically distant organisms. However, it is highly probable that these domains, which are also present in many non-adhesive proteins, were convergently acquired from ancestral proteins with unrelated general functions. We herein present morphological, structural, and molecular convergences between different attachment mechanisms in aquatic animals that likely arose in response to shared functional and selective pressures.
ARTICLE | doi:10.20944/preprints202203.0179.v1
Subject: Biology, Animal Sciences & Zoology Keywords: functional evolution; herpetology; limbless lizards; morphotypes
Online: 14 March 2022 (09:10:31 CET)
Evolutionary ecologists seek to understand how phenotypic evolution relates to population level traits. To document that, we quantified functional traits (escape strategy, sprint speed and predatory performance) and population density across 10 lizard species representing the acquisition of burrowing snake-like morphs (BSLM), from Brazil. We used phylogenetic mixed models to test if: a) morph and substrate affects lizards’ flight strategy and speed, b) BSLM species get more access to different potential prey types than lacertoid ones, when in syntopy, and d) morph is correlated with population abundance and habitat use in a way expected from the output of the previous experiments. We found that BSLM rigidly relied on burrowing as flight strategy, while syntopic lacertoid species changed their strategy according to the substrate. Also, sand has opposite effects on sprint speed depending on morph, making lacertoids run slower and BSLM faster. Despite BSLM were overall slower than lacertoids, they were as good hunters of challengingly fast prey, and better hunters of underground prey. Further, in the sandy habitats where these morphs are syntopic, prey is most available at superficial leaf litter under vegetation, although there is a large stock of underground prey, under the vegetation, too. These results support our observed higher importance of sand for BSLM’s populations and the higher importance of vegetation for lacertoids’ population density. Finally, although BSLM species reached the highest populational densities among the studied species, a systematic effect of morphological evolution on abundance remains elusive.
ARTICLE | doi:10.20944/preprints202203.0120.v1
Subject: Biology, Animal Sciences & Zoology Keywords: magnesium; absorption; rumen; butyrate; volatile fatty acids
Online: 8 March 2022 (08:59:14 CET)
The aim of the present study was to measure the apparent absorption of magnesium (Mg) originating from Mg-butyrate. Six mid-lactation Holstein Friesian dairy cows were used with dietary treatments arranged in a cross-over design. Two different diets were fed during the experiment, consisting of a low Mg diet without Mg-butyrate (L-Mg, 3.1g Mg/kg dry matter) or a high Mg diet with Mg-butyrate (H-Mg, 3.9 g Mg/kg dry matter). Cows offered the L-Mg diet ingested 54.7 g Mg/day while the cows fed the H-Mg diets ingested 66.3 g Mg/day (P < 0.001). The fecal excretion of Mg, however, was similar between the two experimental diets (P = 0.174). Consequently, apparent Mg absorption was found to be 7.9 percentage units greater (P = 0.038) when the cows were fed the diet supplemented with Mg-butyrate. The greater Mg absorption after feeding the H-Mg diet was, however, not reflected by a greater urinary Mg concentration (P = 0.228). These results indicate that the availability of Mg from the Mg-butyrate supplemented diet is high (34.1% of intake). The fractional Mg absorption from Mg-butyrate was calculated to be 71.6%. In conclusion, Mg-butyrate is an attractive alternative to supplement dairy rations with Mg.
HYPOTHESIS | doi:10.20944/preprints202203.0027.v1
Subject: Biology, Animal Sciences & Zoology Keywords: camouflage; colouration; arsenic; toxicity; common river prawn
Online: 1 March 2022 (16:29:06 CET)
Camouflage is an interesting adaptation (for survivability) by organisms in terms of different aggregation or fusion of colourations. Understanding these camouflage strategies in the presence of arsenic on transparent/semi-transparent species is pretty challenging. Previously, several researchers have demonstrated that colouration or pigmentation strategy in an organism is a strategy to merge with the environment to escape from predatory threats. Our study was done on a semi-transparent freshwater prawn species which exhibits a strategy of pigment droplets on its exoskeleton. Unlike previous studies, our findings robustly indicate the fact that pigment droplets are not the only reason for colouration. The pigment droplets rather regulate the darkness of the exoskeleton. However, the transparency of the abdominal muscles additionally plays a crucial role in creating a background of the pigment droplets. The transparency muscles allow light to pass through the abdomen, thus creating a semi-transparent appearance. The degree of semi-transparency is also regulated by the intensity of light. The abdomen and the pigment droplets on the exoskeleton cumulatively as well as contrastingly maintain the transparency and the colour quotient of the prawns. In our study, we have majorly concentrated on the abdominal region of the prawns as it appears to be the key semi-transparent feature of the organism. This research is directed to an ecotoxicological aspect where we showed that arsenic in chronic non-lethal concentration, can notably alter the colouration pattern in this prawn model within a short period of time. Finally, we have used an image processing algorithm to assess the alteration of colouration in this organism.
TECHNICAL NOTE | doi:10.20944/preprints202202.0341.v1
Subject: Biology, Animal Sciences & Zoology Keywords: Ctenophora; breeding; comb jelly; cultivation; husbandry; protocol; sea walnut
Online: 25 February 2022 (15:26:33 CET)
Ctenophores are marine organisms attracting significant attention from evolutionary, molecular biology and ecological research. Here we describe an easy and affordable set-up to maintain a stable culture of the ctenophore Mnemiopsis leidyi. The challenging delicacy of the lobate ctenophores can be met by monitoring the water quality, providing the right nutrition, and adapting the handling and tank set-up to their fragile gelatinous body plan. Following this protocol allows stable laboratory lines, a continuous supply of embryos for molecular biological studies, and independence from population responses to environmental fluctuations.
REVIEW | doi:10.20944/preprints202202.0284.v1
Subject: Biology, Animal Sciences & Zoology Keywords: Recreational fisheries fishery management; introgressive hybridization; stocking; non-native species; allochthonous species; trout fishing; trout taxonomy
Online: 23 February 2022 (03:09:39 CET)
During the last 150 years, the trout-culture industry focused on enhancing trout populations by stocking, in response to the growing anglers’ demand and the habitat degradation associated to the rapid urbanization and hydropower development. The industrialized north of Italy, home to the Italian Alpine and subalpine trout populations, is the source of most of the revenues of the national trout-culture industry. Its rapid growth and the massive introduction of non-native interfertile trouts eroded the genetic diversity of native lineages, leading to harsh confrontations between scientists, institutions, and sportfishing associations. We review here the state of art of the taxonomy and distribution of the northern Italian native trouts, presenting both scientific results and historical documentation. We think the only native trouts in this region are Salmo marmoratus, widespread in this region, plus small and fragmented populations of S. ghigii, present only in the Southwestern Alps. We strongly recommend the interruption of stocking of domesticated interfertile non-native trouts in this area, and recommend the adoption of Evolutionary Significant Units for salmonid fishery management. We further propose future research directions for a sustainable approach to the conservation and ecosystem management of the fishery resources and inland waters of northern Italy.
ARTICLE | doi:10.20944/preprints202202.0191.v1
Subject: Biology, Animal Sciences & Zoology Keywords: Wolbachia; Aedes sp.; Culex sp.; Ctenocephalides felis; Acanthocheilonema reconditum; poly-merase chain reaction
Online: 16 February 2022 (04:38:29 CET)
Background: Wolbachia sp. is an endosymbiotic bacterium that is widespread among arthropods and filarial nematodes. It affects the fertility and maturation of its arthropod hosts, which in some cases are vectors of infectious diseases of humans and animals. Acanthocheilonema reconditum is a canine filarial nematode, which can eventually infect humans through arthropods. An outbreak of A. reconditum has been reported recently in dogs in the southwest of Colombia, specifically in Cali. In addition, Wolbachia sp. is currently used for the biological control of the mosquito Aedes egypti in cities in the north and south-west of Colombia. Aedes egypti is a vector of dengue fever, a major public health disease in tropical and subtropical areas of the world. The objectives of this study were to detect Wolbachia sp. in mosquito and flea samples collected from dogs in southwestern Colombia and to determine the co-infection of Wolbachia sp. and A. reconditum in the recent outbreak of canine filariasis in Santiago de Cali. Methods: A total of 18 mosquitoes of the genera Aedes and Culex and 18 Ctenocephalides felis fleas were collected in Santiago de Cali and nearby municipalities. DNA was extracted and 16S rRNA and ftsZ gene fragments were identified using conventional and nested polymerase chain reaction. In addition, the presence of Wolbachia sp. was analyzed in 55 cases positive for A. reconditum previously detected in the city, using the same molecular techniques. Results: From the analysis of 18 mosquitoes and 18 fleas, using molecular techniques based on the 16S rRNA and ftsZ genes, Wolbachia sp. was detected in all the arthropods analyzed. Wolbachia sp. was not detected in the samples obtained from dogs infected with A. reconditum. Conclusions: Molecular analyses were shown to be effective for the identification of Wolbachia sp. in mosquitoes and fleas but not in samples positive for A. reconditum.
ARTICLE | doi:10.20944/preprints202202.0108.v1
Subject: Biology, Animal Sciences & Zoology Keywords: 3D printing technology; freshwater turtle; Ocadia Spp.; shell wound healing
Online: 8 February 2022 (12:22:31 CET)
Numerous cases and a shortage of resources usually limit wild animal rescue. New technology implemented might save these severely injured wild animals from the situation of euthanasia by easing the requirement of intensive medication. Three-dimensional (3D) technologies provide precise and accurate results that improve the quality of the medical application. These 3D tools have become relatively low-cost and accessible in the past years. In the medical field of exotic animal, turtle shell defect is highly challenging because of inevitable water immersion. This report is the first attempt to apply the combination of 3D scanning, computer-aid design (CAD), and 3D printing to make a protective device that frees the wound from exposure to water or infection sources. The presenting techniques successfully extricate a wild freshwater turtle from an extensive shell defect within a short period. Integration of multiple sciences to 3D technology can provide a facile model for veterinary medical applications.
COMMUNICATION | doi:10.20944/preprints202201.0462.v1
Subject: Biology, Animal Sciences & Zoology Keywords: roe deer; Tick-Borne Encephalitis; neurologic disease; pathology; genetic characterization
Online: 31 January 2022 (13:21:18 CET)
Tick borne encephalitis virus (TBEV) is the causative agent of Tick borne encephalitis in humans, a severe zoonosis occurring in the Paleartic region mainly transmitted through ticks belonging to the genus Ixodes. In Italy, TBEV is restricted to few foci in the north-eastern part of the country. This report describes for the first time a case of clinical TBE in a roe deer, occurred in the Belluno province, Veneto region, an area highly endemic for the presence of the virus. The affected roe deer showed ataxia, staggering movements, muscle tremors and persistent teeth grinding causing hypersalivation. At necropsy, the macroscopic picture was inconclusive. RNA of TBEV was detected by real-time RT-PCR. Phylogenetic analysis revealed a close relationship to TBEV of the European subtype, and 100% similarity with a virus from the bordering Trento Province. The histological examination of the midbrain confirmed the viral etiology and specific immunofluorescence indicated the presence of a Flavivirus infection and characterized the pattern of infection in the neurons. This report underlines for the first time the occurrence of clinical encephalitic manifestations due to TBEV in a roe deer, thussuggesting to include this pathogen in the frame of differential diagnosis in this species.
ARTICLE | doi:10.20944/preprints202201.0295.v1
Subject: Biology, Animal Sciences & Zoology Keywords: WGCNA; lung development; histological structure; molecular changes; amphibian
Online: 20 January 2022 (10:16:10 CET)
Metamorphosis is a critical process for most anurans to transition from water to land. The appearance of air-breathing lungs occurs during the change in oxygen medium from water to air. Revealing the structural construction and molecular switches of lung organogenesis is essential to understand the realization of air-breathing function. In this study, histology and transcriptomics were combinedly conducted to explore these issues in Microhyla fissipes lungs during metamorphosis. During the pro-metamorphic phase, histological structure improving of the alveolar wall was accompanied by robust substrate metabolism and protein turnover. The lungs, at the metamorphic climax phase, are characterized by an increased number of cilia in the alveolar epithelial cells and collagenous fibers in the connective tissues, corresponding to the transcriptional upregulation of cilia and extracellular matrix-related genes. The post-metamorphic lungs strengthen the contracting function, as suggested by the thickened muscle layer and the upregulated expression of genes involved in muscle contraction. The blood–gas barrier is fully developed in adult lungs whose transcriptional features are tissue growth and differentiation regulation and immunity. Importantly, significant transcriptional switches of pulmonary surfactant protein and hemoglobin facilitate air-breathing. Our results illuminated four key steps of lung development for amphibians to transition from water to land.
ARTICLE | doi:10.20944/preprints202201.0251.v1
Subject: Biology, Animal Sciences & Zoology Keywords: eIF2α; eIF2α-kinases; diets; stressors; Rachycentron canadum; cobia cells
Online: 18 January 2022 (10:39:48 CET)
The present study investigates the response of the marine fish cobia, Rachycentron canadum, to stressors as measured by phosphorylation of the α-subunit of the translational initiation factor, eIF2. eIF2α is the target of phosphorylation by a family of kinases that respond to a range of physiological stressors. Phosphorylation of eIF2α not only inhibits overall protein synthesis, but allows cells to reprogram gene expression to adapt to, and recover from, stress. The deduced coding sequence of cobia eIF2α has 94 % identity to both zebrafish (Danio rerio) and human eIF2α sequences with identical phosphorylation and kinase docking sites. The present study uses a cobia cell line, Cm cells, derived from muscle, as well as cobia larvae to investigate the response of cobia eIF2α to various stressors. In Cm cells, phosphorylation of eIF2α is increased by nutrient deficiency, leucinol, and ER stress, consistent with the activation of the eIF2 kinases, GCN2, and PERK. In cobia juveniles, diet and water temperature affect the phosphorylation state of eIF2α. We conclude that evaluation of eIF2α phosphorylation could function as an early marker to evaluate diet, aquaculture stressors and disease in cobia and may be of particular use in the optimization of conditions for raising cobia larvae and juveniles.
HYPOTHESIS | doi:10.20944/preprints202201.0097.v1
Subject: Biology, Animal Sciences & Zoology Keywords: Stem cells; proliferation; tumour; benign tumour; cancer
Online: 10 January 2022 (11:05:50 CET)
Our tissues usually have just the right number of cells to optimally fulfil their function. Not enough cells within a tissue can lead to dysfunction, while too many cells result in a tumour. Yet, how this homeostatic balance is maintained remains poorly defined. Most differentiated cells within tissues have a finite lifespan and need to be replaced at a corresponding pace to maintain tissue homeostasis. These new differentiated cells are generated by proliferation of the stem/progenitor cells that serve the tissue. Work in simple invertebrates clearly suggests stem cells respond to at least two types of signals: niche signaling and growth factors. Niche signals promote the undifferentiated state by preventing differentiation, and thus allow for stem cell self-renewal. Growth factor sources comprise a systemic input reflecting the animal’s nutritional status, and a localized, homeostatic feedback from the tissue that the stem cells serve. That homeostatic signal couples stem cell proliferation rates to the tissue’s need for new differentiated cells. Evidence from simple organisms suggests two types of benign tumours can arise from deregulation of either niche or homeostatic signaling. Namely, constitutive niche signaling promotes the formation of undifferentiated “stem cell” tumours, while defective homeostatic signaling leads to the formation of differentiated tumours. We propose that these principles may be conserved and underlie benign tumour formation in humans, while benign tumours can evolve into cancer.
REVIEW | doi:10.20944/preprints202112.0416.v1
Subject: Biology, Animal Sciences & Zoology Keywords: macrophage, bistability, metabolism, systems biology, miRNA
Online: 25 December 2021 (00:02:25 CET)
Macrophages are essential innate immune cells characterized by a high diversity and plasticity. In vitro, their full dynamic range of activation profiles include the classical pro-inflammatory (M1) and the alternative anti-inflammatory (M2) program. Bistability usually arises in biological systems that contain a positive-feedback loop or a mutually inhibitory, double-negative-feedback loop, which are common regulatory motifs reported for macrophage transitions from one activation state to the other one. This switch-like behavior of macrophage is observed at four different levels. First, a decision-making module in signal transduction includes mutual inhibitory interactions between M1 (STAT1 and NF-KB/p50-p65) and M2 (STAT3 and NF-KB/p50-p50) signaling pathways. Second, a switch-like behavior at the gene expression level includes complex network motifs of transcription factors and miRNAs. Third, those changes impact metabolic gene expression leading to several switches in energy production, NADPH and ROS production, TCA cycle functionality, biosynthesis and nitrogen metabolism. Fourth, metabolic changes are monitored by specialized metabolic sensors coupled to AMPK and mTOR activity to provide stability by maintaining the signals to promote either M1 or M2 activation. The targeting of robust molecular switches has the potential to treat a broad range of widespread diseases such as sepsis, cancer or chronic inflammatory diseases.
BRIEF REPORT | doi:10.20944/preprints202112.0368.v1
Subject: Biology, Animal Sciences & Zoology Keywords: Raccoon dog parvovirus; Epidemiology; VP2 gene; NS1 gene; Evolutionary analysis.
Online: 22 December 2021 (12:52:50 CET)
To understand the epidemiological status of parvovirus (RDPV) in raccoon dogs, intestinal tissues of raccoon dogs in Liaoning Province of China were collected and evaluated. Three strains of raccoon dog parvovirus were successfully isolated from 12 intestinal tissues. Nine samples were positive for RDPV, with a positive rate of 75%. The VP2 and NS1 genes of the viruses were cloned and subjected to sequencing for analysis. The nucleotide sequences of the VP2 gene showed 99.94% similarity to the CPV-2a/Racoon dog/QHD/2/19(MT183665) strain, and the nucleotide sequences of the NS1 gene showed 99.75% similarity to RDPV-DP1 NS1(MF996335) strain. The three isolates belonging to the CPV-2a cluster were further confirmed by amino acid sequence alignment and phylogenetic analysis. Our study enriched the epidemiological data of parvovirus in raccoon dogs in the investigating region, and the results will be helpful for future investigation of the variations and transmission of raccoon dog parvoviruses.
REVIEW | doi:10.20944/preprints202112.0027.v2
Subject: Biology, Animal Sciences & Zoology Keywords: Zoo animal welfare; Five Domains; Validity; Animal-based; Resource-based; Scoring
Online: 22 December 2021 (11:59:32 CET)
Zoos are increasingly putting in place formalized animal welfare assessment programs to allow monitoring of welfare over time, as well as to aid in resource prioritization. These programs tend to rely on assessment tools that incorporate resource-based and observational animal- focused measures since it is rarely feasible to obtain measures of physiology in zoo-housed animals. A range of assessment tools are available which commonly have a basis in the Five Domains framework. A comprehensive review of the literature was conducted to bring together recent studies examining welfare assessment methods in zoo animals. A summary of these methods is provided with advantages and limitations of the approach es presented. We then highlight practical considerations with respect to implementation of these tools into practice, for example scoring schemes, weighting of criteria, and innate animal factors for consideration. It is concluded that would be value in standardizing guidelines for development of welfare assessment tools since zoo accreditation bodies rarely prescribe these. There is also a need to develop taxon or species- specific assessment tools to inform welfare management.
ARTICLE | doi:10.20944/preprints202112.0322.v1
Subject: Biology, Animal Sciences & Zoology Keywords: Development; area; livestock; beef cattle; corporation
Online: 21 December 2021 (11:31:54 CET)
Livestock is an integral part of agriculture which significantly contributes to the economic and socio-economic development. Based on its potential in natural resources and human resources, East Kalimantan Province has opportunity to become a beef cattle development area. The development of a corporate-based beef cattle breeding area is an approach which taken toward industrial and business-oriented beef cattle breeding. The concept of breeder corporation will create new strengths such as strengths in human resources, capital, and banking in business development, which can more open the opportunities for the success and growth of the breeder's business. The development of a corporate-based beef cattle breeding area provides opportunities, including: 1) improving the competitiveness and added value of the region and beef cattle commodities in order to support national sustainable food security; 2) strengthening the livestock business system in one area management in a holistic manner; and 3) strengthening breeders institutions in accessing information, technology, public facilities and infrastructure, capital, processing and marketing, so that the concept is expected to be applied in East Kalimantan Province.
ARTICLE | doi:10.20944/preprints202112.0166.v1
Subject: Biology, Animal Sciences & Zoology Keywords: methyl sulfonyl methane; sodium sulfate; laying hen; antioxidant capacity
Online: 10 December 2021 (08:32:41 CET)
The present study was conducted to investigate the comparative effects of organic and inorganic forms of sulfur, methyl sulfonyl methane (MSM) and sodium sulfate (SS), on laying performance, egg quality, ileal morphology, ileal volatile fatty acids, and antioxidant and stress markers in various biological samples in aged laying hens. A total of 144, 73-week-old Lohman Brown-Lite laying hens were randomly assigned to one of three experimental diets: basal diet (CONT), CONT + 0.2% MSM (MSM), and CONT + 0.3% SS (SS). The trial lasted for 12 weeks. MSM and SS groups contained 0.07% of sulfur, either organic or inorganic. Dietary MSM did not affect egg production and feed conversion ratio at 12 weeks compared with the CONT group. Dietary sulfur did not affect egg quality except for Haugh unit at 4 weeks which was lowered (P < 0.05) in the SS group. Compared with the CONT group, higher (P < 0.05) villus height and crypt depth ratio was observed in the SS group. None of dietary sulfur affected the percentages of short-chain fatty acids in the ileum. Total antioxidant capacity of liver increased (P < 0.05) in laying hens fed MSM- and SS-added diets compared with the CONT group. The MSM and SS groups lowered (P < 0.05) malondialdehyde (MDA) concentration in serum samples compared with the CONT. Finally, dietary MSM had the lowest (P < 0.05) MDA concentrations in yolk samples. Taken together, our study showed that dietary organic and inorganic sulfur have positive effects on ileal morphology and antioxidant capacity in laying hens. However, SS-mediated inhibition in laying performance needs to be clarified.
ARTICLE | doi:10.20944/preprints202112.0108.v1
Subject: Biology, Animal Sciences & Zoology Keywords: cnidarian venom; cnidocyst discharge; cnidocyte; ocean literacy; risk prevention; seawater; sting; vinegar
Online: 7 December 2021 (14:43:25 CET)
Anemonia viridis is an abundant and widely-distributed temperate sea anemone that can form dense congregations of individuals. Despite the potential severity of its sting, few detailed cases have been reported. We report a case of a severe toxic reaction following an A. viridis sting in a 35-year-old oceanographer. She developed severe pain, itching, redness and burning sensation, which worsened one week after treatment with anti-inflammatories, antihistamines and corticosteroids. Prompted by this event, and due to the insufficient risk prevention, lack of training for marine-environment users and lack of research into sting-specific first-aid protocols, we evaluated the cnidocyst response to five different compounds commonly recommended as rinse solutions in first-aid protocols (seawater, vinegar, ammonia, baking soda and freshwater) by means of the Tentacle Solution Assay. Vinegar and ammonia triggered an immediate and massive cnidocyst discharge after their application and were classified as activator solutions. Baking soda and freshwater were also classified as activator solutions, although with a lower intensity of discharge. Only seawater was classified as a neutral solution and therefore recommended as a rinse solution after A. viridis sting, at least until an inhibitory solution is discovered.
ARTICLE | doi:10.20944/preprints202112.0082.v1
Subject: Biology, Animal Sciences & Zoology Keywords: Avian olfaction; foraging; herbivore-induced plant volatiles; defence against herbivory
Online: 6 December 2021 (15:21:49 CET)
The tri-trophic interactions between plants, insects, and insect predators and parasitoids are often mediated by chemical cues. The attraction to Herbivore-Infested Plant Volatiles (HIPVs) has been well documented for arthropod predators and parasitoids, and more recently for insectivorous birds. The attraction to plant volatiles induced by the exogenous application of methyl jasmonate (MeJA), a phytohormone typically produced in response to an attack of chewing herbivores, have provided controversial results, both in arthropod and avian predators. In this study, we aimed to examine whether potential differences in the composition of bouquets of volatiles produced by Herbivore-infested and MeJA-treated Pyrenean oak trees (Quercus pyrenaica) were related to differential avian attraction, as results from a previous study suggested. Results showed that the overall emission of volatiles produced by MeJA-treated and Herbivore-infested trees did not differ, and were higher than emissions of Control trees, although MeJA treatment showed more significant reaction and released several specific compounds in contrast to Herbivore-induced trees. These slight differences in the volatile composition may explain why avian predators were not so attracted to MeJA-treated trees as observed in a previous study in this plant-herbivore system. Unfortunately, the lack of avian visits to the experimental trees in the current study does not allow us to confirm this result and points out the need to perform more robust predator studies.
ARTICLE | doi:10.20944/preprints202111.0526.v1
Subject: Biology, Animal Sciences & Zoology Keywords: VP28; WSSV; real-time PCR; viral load; apoptosis
Online: 29 November 2021 (11:55:17 CET)
White Spot Syndrome Virus (WSSV) has emerged as one of the most prevalent and lethal viruses globally, and infects both shrimps and crabs in the aquatic environment. This study aimed to investigate the occurrence of WSSV in different ghers of Bangladesh and the virulence of the circulating phylotypes. We collected 360 shrimp (Penaeus monodon) and 120 crab (Scylla sp.) samples from the South-East (Cox’s Bazar) and South-West (Satkhira) coastal regions of Bangladesh. The VP28 gene-specific PCR assays and sequencing revealed statistically significant (p < 0.05, Kruskal Wallis test) differences in the prevalence of WSSV in shrimps and crabs between the study areas (Cox’s Bazar and Satkhira), and over the study periods (2017-2019). The mean Log load of WSSV varied from 8.40 (Cox’s Bazar) to 10.48 (Satkhira) per gram of tissue. The mean values for salinity, dissolved oxygen, temperature and pH were 14.71±0.76 ppt, 3.7±0.1 ppm, 34.11±0.38˚C and 8.23±0.38, respectively in the WSSV-positive ghers. The VP28 gene-based phylogenetic analysis showed an amino-acid substitution (E→G) at 167th position in the isolates from Cox’s Bazar (referred to as phylotype BD2) compared to the globally circulating one (BD1). Shrimp PL artificially challenged with BD1 and BD2 phylotypes with filtrates of tissue containing 0.423 X 109 copies of WSSV per mL resulted a median LT50 value of 73 hrs and 75 hrs, respectively. The in-vivo trial showed higher mean Log WSSV copies (6.47±2.07 per mg tissue) in BD1 challenged shrimp PL compared to BD2 (4.75±0.35 per mg tissue). Crabs infected with BD1 and BD2 showed 100% mortality within 48 hrs and 62 hrs of challenge, respectively with mean Log WSSV copies of 12.06±0.48 and 9.95±0.37 per gram tissue, respectively. Moreover, shrimp antimicrobial peptides (AMPs) penaeidin and lysozyme expression was lower in BD1 challenged group compared to BD2 challenged shrimps. These results collectively demonstrated that relative virulence properties of WSSV based on mortality rate, viral load and expression of host immune genes in artificially infected shrimp PL could be affected by single aa substitution in VP28.
ARTICLE | doi:10.20944/preprints202111.0436.v1
Subject: Biology, Animal Sciences & Zoology Keywords: Prophase I of meiosis; chromatin; synaptonemal complex; chromosome; satellite DNA; chromocenter; nuclear architecture; MSCI; sex chromosomes; interactions of non-homologous chromosomes; chromatin silencing
Online: 23 November 2021 (15:58:17 CET)
Pericentromeric regions of chromosomes enriched in tandemly repeated satellite DNA although representing a significant part of eukaryotic genomes are still understudied mainly due to interdisciplinary knowledge gaps. Recent studies suggest their important role in genome regulation, karyotype stability and evolution. Thus, the idea of satellite DNA as a junk part of the genome was refuted. Integration of data about molecular composition, chromosome behaviour and details of in situ organization of pericentromeric regions is of great interest. The objective of this work was a cytogenetic analysis of the interactions of pericentromeric regions non-homologous chromosomes in mouse spermatocytes using immuno-FISH. We analysed two events: the associations between cerntomeric regions of X chromosome and autosomes, and associations between centromeric regions of autosomal bivalents forming chromocenters. We conclude that X chromosome form temporary synaptic associations with different autosomes in early meiotic prophase I which normally can be found at pachytene-diplotene without signs of pachytene arrest. These associations are formed between the satellite DNA-enriched centomeric regions of X chromosome and different autosomes but not involve the satellite-poor centromeric region of Y-chromosome. We suggest the mechanism of X chromosome competitive replacement from such associations during synaptic correction. We showed that centromeric region of the X chromosome remains free of γH2Ax-dependent chromatin inactivation, while Y chromosome is completely inactivated. This findings highlights the predominant role of associations between satellite DNA-enriched regions of different chromosomes including X. We assume that X-autosome temporary associations is a manifestation of an additional synaptic disorders checkpoint. These associations are normally corrected before the late diplotene. We revealed that the intense spreading conditions applied to the spermatocytes I nuclei did not lead to destruction of stretched chromatin fibers i.e. elongated chromocenters enriched in satellite DNA. Revealed by us tight associations between pericentromeric regions of different autosomal bivalents and X chromosome may represent the basis for repeat stability maintenance in autosomes an X chromosome. The consequences of our findings are discussed. We obtained the preparations of mouse spermatocytes nuclei in the meiotic prophase I using two approaches: standard and extremely intense surface spread techniques. Using immuno-FISH we visualized tandemly repeated mouse Major and Minor satellite DNA located in the pericentromeric regions of chromosomes and performed a morphological comparison of the standard- and intensely spreaded meiotic nuclei. Based on our results, we assume the remarkable strength of the chromocenter-mediated associations, “chromatin “bridges”, between different bivalents at the pachytene and diplotene stages. We have demonstrated that the chromocenter “bridges” between the centromeric ends of meiotic bivalents are enriched in both tandemly repeated Major and Minor satellite DNA. Association of centromeric regions of autosomal bivalents and X-chromosome but not with Y-chromosome correlates with the absence of Major and Minor satellites on Y-chromosome. We suggest that revealed tight associations between pericentromeric regions of bivalents may represent the network-like system providing dynamic stability of chromosomal territories, as well as add new data for the hypothesis of ectopic recombination in these regions which supports sequence homogeneity between non-homologous chromosomes and does not contradict the meiotic restrictions imposed by the crossing-over interference near centromeres. We conclude that nuclear architecture in meio-sis may play an essential role in contacts between the non-homologous chromosomes providing the specific characteristics of pericentromeric DNA.
ARTICLE | doi:10.20944/preprints202111.0352.v1
Subject: Biology, Animal Sciences & Zoology Keywords: skeletal abnormalities; vertebral column; quality index; body shape; finfish aquaculture
Online: 19 November 2021 (12:43:54 CET)
The lordosis of the haemal part of the vertebral column is a frequent abnormality in reared fish. Haemal lordosis develops during the late larval and early juvenile period, mainly due to the high swimming activity of the fish in the rearing tanks. In the present study we examined whether haemal lordosis recovers during the growth of European sea bass. Furthermore, we aimed to develop simple morphometric indices (PrAn1 and PrAn2) that could link the severity of lordosis at the juvenile stage with fish morphological quality at harvesting. At 111 days post-hatching (dph, 53±4 mm standard length, SL), 600 seabass juveniles with lordotic (L, 200 fish) or normal (N, 400 fish) external morphology were selected and introduced in a common tank. At 150 dph (75±7 mm SL), 350 fish were randomly selected, pit-tagged and transferred in a sea cage for on-growing up to 502 dph (234±16 mm SL). The morphological examination of the fish at 150 and 502 dph revealed that the 60% (46 out of 77) of L juveniles turned into normal phenotype by the end of on-growing period. Interestingly, 56% of the fish with recovered external morphology (N-Rec) presented either a completely normal vertebral column (31%) or minor abnormalities of individual vertebrae (25%). Following the results of geometric morphometric analysis, the differences in the body shape between N-Rec and N fish were not significant (p>0.05, canonical variate analysis). The examined morphometric indices were effective in discriminating the normal fish from the 58% (PrAn1) to 65% (PrAn2) of lordotic juveniles. Results are discussed with respect to the mechanism of lordosis recovery, as well as to their application for the quality control and cull out of the abnormal fish in the commercial hatcheries.
REVIEW | doi:10.20944/preprints202111.0279.v1
Subject: Biology, Animal Sciences & Zoology Keywords: animal model; dietary components; feeding strategy; gene expression; lipid profile; nutrients; nutrigenomic; physiological processes; signaling pathways; transcription.
Online: 16 November 2021 (09:07:22 CET)
Studies on the influence of dietary components and their effects are fundamental for nutrigenomics, or the study of how nutrients can be cellular sensors, how they affect biological processes and gene expression in different tissues. Lipids are an important source of fatty acids (FA) and energy and are fundamental to biological processes and influence the regulation of transcription. Pigs are excellent model to study nutrigenomics, particularly lipid metabolism because the deposition and composition of FA in their tissues reflect the composition of FA in their diet. Recent studies show that FA supplementation is important in production systems, such as growing and finishing pigs, as it can improve the energy value of the feed, help reduce costs, improve animal welfare, and influence the nutritional value of the meat. Studies show that oleic (OA), linoleic (LA), docosahexaenoic (DHA), and eicosapentaenoic (EPA) acids are associated with the regulation of transcription in tissues such as muscle, liver, adipose tissue, and brain. Other studies indicate that EPA and DHA are associated with changes in specific signaling pathways, altering gene expression and biophysical properties of membranes. This review, therefore, focuses on the current knowledge of the effects of dietary FA on production traits and gene expression.
ARTICLE | doi:10.20944/preprints202111.0258.v1
Subject: Biology, Animal Sciences & Zoology Keywords: feral cat; Felis catus; Australia; Indigenous Protected Area; 1080; camera; tracking
Online: 15 November 2021 (11:57:03 CET)
Feral cats are both difficult to manage and harder to monitor. We analysed the cost-efficacy of monitoring the pre- and post-bait abundance of feral cats via camera-traps or track counts using four years of data from the Matuwa Indigenous protected Area. Additionally, we report on the recovery of the feral cat population and the efficacy of subsequent Eradicat® aerial baiting programs following 12 months of intensive feral cat control in 2019 that consisted of aerial baiting and leg-hold trapping. Significantly fewer cats were captured in 2020 (n = 8) compared to 2019 (n = 126). Pre-baiting surveys for 2020 and 2021 suggested that the population of feral cats on Matuwa was very low, at 5.5 and 4.4 cats/100 km respectively, which is well below our target threshold of 10 cats/100 km. Post-baiting surveys then recorded 3.6 and 3.0 cats/100 km respectively, which still equates to a 35% and 32% reduction in cat activity. Track counts recorded significantly more feral cats than camera traps and were cheaper to implement. We recommend that at least two methods of monitoring cats be implemented to prevent erroneous conclusions.
ARTICLE | doi:10.20944/preprints202111.0140.v1
Subject: Biology, Animal Sciences & Zoology Keywords: Streptococcus uberis; mastitis; typing; antimicrobial susceptibility; resistance genes
Online: 8 November 2021 (13:12:44 CET)
Intrammary infections are a major problem for dairy sheep farms, and Streptococcus uberis is one of the main etiological agents of ovine mastitis. Surveys on antimicrobial resistance are still limited in sheep and characterization of isolates is important for acquiring information on resistance and for optimizing therapy. In this study, a sampling of 124 S. uberis isolates collected in Sardinia (Italy) from sheep milk was analysed by multilocus-sequence typing (MLST) and pulsed field gel electrophoresis (PFGE) for genetic relatedness. All isolates were also subjected to antimicrobial susceptibility analysis by the disk diffusion test using a panel of 14 antimicrobials. Resistance genes were detected by PCR assays. MLST analysis revealed that the isolates were grouped into 86 sequence types (STs), of which 73 were new genotypes, indicating a highly diverse population of S. uberis. The most frequently detected lineage was the clonal complex (CC)143, although representing only 13.7% of all characterized isolates. A high level of heterogeneity was also observed among the SmaI PFGE profiles, with 121 unique patterns. Almost all (96.8%) isolates were resistant to at least one antimicrobial, while all exhibited phenotypic susceptibility to oxacillin, amoxicillin-clavulanic acid and ceftiofur. Of the antimicrobials tested, the highest resistance rate was found against streptomycin (93.5%), kanamycin (79.8%) and gentamicin (64.5%), followed by novobiocin (25%) and tetracycline-TE (19.3%). Seventy-four (59.7%) isolates were simultaneously resistant to all aminoglycosides tested. Seventeen isolates (13.7%) exhibited multidrug resistance. All aminoglycosides-resistant isolates were PCR negative for aad-6 and aphA-3’ genes. Among the TE-resistant isolates, the tetM gene was predominant, indicating that the resistance mechanism is mainly mediated by the protection of ribosomes and not through the efflux pump. Three isolates were resistant to erythromycin, and two of them harboured the ermB gene. This is the first study reporting a detailed characterization of the S. uberis strains circulating in Sardinian sheep. Further investigations will be needed to understand the relationships between S. uberis genotypes, mastitis severity, and intra-mammary infection dynamics in the flock, as well as to monitor the evolution of antimicrobial resistance.
REVIEW | doi:10.20944/preprints202111.0127.v1
Subject: Biology, Animal Sciences & Zoology Keywords: Apoptosis; preclinical research; diabetes type 2; HIPPO pathway
Online: 8 November 2021 (11:45:49 CET)
Diabetes mellitus is a heterogeneous disease of complex etiology and pathogenesis. Hyperglycemia leads to many serious complications, but also directly initiates the process of β cell apoptosis. A potential strategy for the preservation of pancreatic β cells in diabetes may be to inhibit the implementation of pro-apoptotic pathways or to enhance the action of pancreatic protective factors. The HIPPO signaling pathway is proposed and selected as a target to manipulate the activity of its core proteins in therapy - basic research. MST1 and LATS2 as major upstream signaling kinases of the Hippo pathway are considered as target candidates for pharmacologically induced tissue regeneration and inhibition of apoptosis. Manipulating the activity of components of the HiPPO pathway offers a wide range of possibilities, and thus is a potential tool in the treatment of diabetes and the regeneration of β cells. Therefore, it is important to fully understand the processes involved in apoptosis in diabetic states and to fully characterize the role of this pathway in diabetes. Therapy consisting in slowing down or stopping the mechanisms of apoptosis may be an important direction of diabetes treatment shortly.
ARTICLE | doi:10.20944/preprints202111.0094.v1
Subject: Biology, Animal Sciences & Zoology Keywords: Sparus aurata; social stress; hierarchy; cortisol; phagocytosis
Online: 4 November 2021 (08:44:54 CET)
Abstract: Social stress can affect the ability of the fish to respond to various stressors, such as pathogens or environmental variations. In this paper, the effects of social stress on gilt-head bream (Sparus aurata) were investigated. To study the effects of physiological stress, we evaluated biochemical and cellular parameters as cortisol, glucose, lactate, osmolarity and phagocytosis 24 hours after the establishment of social hierarchy. Social hierarchy was determined and characterised by behavioural observation (aggressive acts and feeding order) of the specimens (dominant “α”, subordinate “β” and “γ”). After the establishment of the social hierarchy, we observed that the levels of plasma cortisol and other biochemical stress markers (glucose and lactate) were higher in subordinate individuals than in dominant ones. In addition, the modulation of phagocytic activity of the peritoneal cavity cells (PEC) demonstrated that social stress appeared to affect the immune response. At last, principal component analysis clearly separated the subordinate fish groups from the dominant groups based on stress markers and phagocytic activity of the peritoneal exudates cells.
ARTICLE | doi:10.20944/preprints202111.0033.v1
Subject: Biology, Animal Sciences & Zoology Keywords: Africa; biodiversity infrastructure; Clupeidae; Clupeiformes; Dactylogyridea; flatworm; historical collection; Monogenea; Pellonulini; sardine
Online: 2 November 2021 (10:28:05 CET)
Unlike their marine counterparts, tropical freshwater clupeids receive little scientific attention. However, they sustain important fisheries that may be of (inter)national commercial interest. Africa harbours over 20 freshwater clupeid species within Pellonulini. Recent research suggests their most abundant parasites are gill-infecting monogenean flatworms within Kapentagyrus. After inspecting specimens of 12 freshwater clupeids from West and Central Africa, mainly sourced in biodiversity collections, we propose 11 new species of Kapentagyrus which we describe using their haptoral and genital morphology. Because of their high morphological similarity, species delineation relies mostly on morphometrics of anchors and hooks. Specifically, earlier, molecular taxonomic work indicated that the proportion between the length of the anchor roots, and between hook and anchor length, are diagnostic. On average, about one species of Kapentagyrus exists per pellonuline species, although Pellonula leonensis harbours four species and Microthrissa congica two, while Microthrissa moeruensis and Potamothrissa acutirostris share a gill monogenean species. This study more than quadruples the number of known species of Kapentagyrus, also almost quadrupling the number of pellonuline species of which monogeneans are known. Since members of Kapentagyrus are informative about their hosts’ ecology, evolutionary history, and introduction routes, this enables a parasitological perspective on several data-poor African fisheries.
ARTICLE | doi:10.20944/preprints202110.0281.v1
Subject: Biology, Animal Sciences & Zoology Keywords: selective pressures; mitochondrial protein-coding genes; subterranean voles; adaptations; subterranean lifestyle.
Online: 19 October 2021 (15:01:42 CEST)
The current study evaluates the selection signals in the evolution of mitochondrial DNA of voles, subfamily Arvicolinae, during the colonization of subterranean environments. The comparative sequence analysis of mitochondrial protein-coding genes of eight subterranean vole species (Prometheomys schaposchnikowi, three species of the genus Ellobius: E. talpinus, E. fuscocapillus and E. lutescens, two species of the genus Terricola: T. subterraneus and T. daghestanicus, Lasiopodomys mandarinus and Hyperacrius fertilis) and their closest aboveground relatives using codon-substitution models was applied. The highest number of selection signatures was detected in genes ATP8 and CYTB. The relaxation of selection was observed in most mtDNA protein-coding genes. In mole voles (genus Ellobius) the signatures of adaptive evolution of mitochondrial genes related to subterranean niche were most pronounced. The number of selection signatures was found to be independent of the evolutionary age of the lineage but fits the degree of specialization to the subterranean niche. The common trends of selective pressures were observed among the evolutionary ancient and highly specialized subterranean rodent families and phylogenetically young lineages of voles. It suggests that the signatures of adaptations in individual mitochondrial protein-coding genes associated with the colonization of the subterranean niche may appear within a rather short evolutionary timespan.
ARTICLE | doi:10.20944/preprints202110.0250.v1
Subject: Biology, Animal Sciences & Zoology Keywords: non-tuberculous mycobacteria (NTM); mycobacteria other than tuberculosis (MOTT); saprophytic environmental mycobacteria; risk groups of microorganisms; Microchiroptera
Online: 18 October 2021 (14:54:06 CEST)
A total of 281 guano samples were collected from caves (N = 181) in 8 European countries (Bulgaria, Czech Republic, France, Hungary, Italy, Romania, Slovakia and Slovenia) and attics in the Czech R. (N = 100). The correlation of detection of mycobacteria between Ziehl-Neelsen (ZN) microscopy and culture examination and qPCR was strong. ZN microscopy was positive in guano from caves (58.6%) more than double than positivity in guano from attics (21.0%; P <0.01). From 89 mycobacterial isolates (73 isolates from cave guano and 16 isolates from attics’ guano) 68 (76.4%) isolates of 19 sp., ssp. and complex were identified as members of 3 Groups (M. fortuitum, M. chelonae, and M. mucogenicum), and 4 Complexes (M. avium, M. terrae, M. vaccae, and M. smegmatis). A total of 20 isolates (22,5%) belonged to risk group 1 (environmental saprophytes), 48 isolates (53.9%) belonged to risk group 2 (potential pathogens), and none of isolates belonged to risk group 3 (obligatory pathogens). When comparing bat guano collected from caves and attics, differences (P<0.01; Mann-Whitney test) were observed for the electrical conductivity, total carbon, total organic and total inorganic carbon. None difference (P> 0.05; Mann-Whitney test) was found for pH and oxidation-reduction potential parameters.
Online: 14 October 2021 (10:33:01 CEST)
The health status and feed conversion efficiency of farmed fish may vary according to management and production methods. Successful aquaculture requires safeguarding the health of the growing fish and optimizing the feed conversion and therefore achieving better FCE thus reducing the amount of feed required to produce farmed fish, reducing the environmental impact generated by fish feed production and reducing aquaculture wastes generated by feed wasted or poorly digested. The present review presents illustrative examples from freshwater aquaculture that suggests the potential dual benefits of focusing on the link between feed conversion and the environmental impact of fresh water fish farms. Apart from the need to support future research on new diets for farmed fish (which is mainly driven by limits in the supply of fish protein and the results price fluctuation of all ingredients used by the aquaculture, feed industry), major improvements can be expected by optimizing feeding regimes and the application of probiotics. Aside from the economic benefits and increased production of fish farms, improved feeding regimes and probiotics are expected to have a significant impact on the welfare of farmed fish as well as on digestion efficiency and the environmental impact of fresh water fish farms.
REVIEW | doi:10.20944/preprints202110.0200.v1
Subject: Biology, Animal Sciences & Zoology Keywords: Newcastle disease; poultry; Pakistan; vaccine; economic affects
Online: 13 October 2021 (11:59:23 CEST)
The poultry industry is affected by many epidemics and Newcastle Disease (ND) is a constant threat, known as a devastating disease for poultry farmers around the world. According to the average death time of chicken embryos, virus strains can be classified as lentogenic, mesogenic, or velogenic. The current research will clarify the vulnerable host range as well as the epidemiology and geographic distribution of ND in Pakistan. The introduction of the virus into poultry can have serious economic consequences, including the loss of production of sick and dying poultry, the cost of control measures (such as population reduction and disinfection measures), and possible trade restrictions in the event of an outbreak. The virus is transmitted by direct contact with sick poultry or carriers. Infected birds can also spread the virus in their feces. It can also be spread through respiratory secretions, contaminated feed, equipment, water, or feces. We will also discuss vaccines that which vaccines are available for NDV in Pakistan and vaccines can fight against this disease or not? In this study, a qualitative risk analysis was carried out to assess Pakistan's vulnerability to the introduction of virulent NDV strains
BRIEF REPORT | doi:10.20944/preprints202110.0173.v1
Subject: Biology, Animal Sciences & Zoology Keywords: Cortisol; Faeces; Glucocorticoids; Iterative baseline; Joey; Koala; Stress
Online: 12 October 2021 (10:18:35 CEST)
Koala (Phascolarctos cinereus) is an iconic folivorous marsupial native to the sclerophyll forests and woodlands of Australia. Due to the ever-changing habitat, this species is highly vulnerable to anthropogenic factors such as habitat loss and fragmentation, and this is reflected in the increasing number of injured and/or diseased koalas over the years. The majority of adult koalas admitted at wildlife hospitals are deceased, either due to natural causes, or have to be euthanized. Thus, orphaned koala joeys constitute a substantial number of wildlife rescues, and mortality is also prevalent in koala joeys being hand-reared/rehabilitated, with little knowledge about the causes of such high rates of mortality. Wildlife hospitals/rehabilitation centres are inherently stressful, and although the hypothalamic–pituitary–adrenal (HPA) axis plays a vital role in mediating the stress endocrine function (by producing glucocorticoids such as cortisol), there are no studies quantifying glucocorticoids in koala joeys. To contribute to this dearth of research, we sampled a total of seven individuals residing at Port Macquarie Koala Hospital and noted their clinical information. Faecal samples were collected from all seven koala joeys during routine cage cleaning. In total, 123 faecal samples were collected, processed and analysed for cortisol using enzyme-immunoassay (EIA). We used the iterative baseline approach to determine baseline and peak concentrations of FCM in koala joeys. Baseline concentrations ranged between 14.11 ng/g – 51.10 ng/g (healthy – sick), whereas, peak FCM concentrations ranged between 25.65 ng/g – 56.58 ng/g (healthy – sick). There was a significant difference (p < 0.05) between FCM concentrations of healthy and impaired individuals. Healthy individuals displayed relatively consistent FCM concentrations, whereas, diseased individuals displayed a significant increase in FCM concentrations over time. Our study provides the first record of baseline and peak FCM concentrations in rescued koala joeys with their associated clinical condition. Future studies can use the iterative baseline approach to determine FCM concentration in wild koala joeys that can serve as a baseline to compare glucocorticoid levels of rescued joeys.
ARTICLE | doi:10.20944/preprints202110.0051.v1
Subject: Biology, Animal Sciences & Zoology Keywords: Longman’s beaked whale; Indopacetus pacificus; Mesoplodon spp.; tuna gillnet fishery; bycatch; citizen science; Arabian Sea
Online: 4 October 2021 (11:40:47 CEST)
Beaked whales (Ziphiidae) are rarely reported in the Arabian Sea. Four new cases (five individuals) were documented in deep waters offshore Pakistan through a pilot programme in 2015-2018 where trained fishers video-recorded net entanglements in the pelagic tuna drift gillnet fishery. Videos were analysed frame-by-frame. The large body size (est. 5-6m) of one specimen, its moderately bulbous melon, long tubular rostrum and a large falcate dolphin-like dorsal fin, indicated Longman’s beaked whale Indopacetus pacificus. It represents the first record for Pakistan (EEZ), and with a stranding at Gujarat, India, a second for the northern Arabian Sea. The other 4 ziphiids were significantly smaller (est. 3– 4.5m), with a decidedly non-bulbous melon, variable short to moderately short rostra, falcate to subtriangular dorsal fin and a nondescript greyish colouration, identified as Mesoplodon spp. Video quality was poor but none of the specimens showed tusks, arched mandible lines or noticeable linear tooth rakes, practically excluding adult males. The successful release of all net-entangled beaked whales is unprecedented. The simultaneous bycatch of two mesoplodonts in the same net set is equally exceptional. This citizen science strategy adds to our understanding of the distribution of I. pacificus and mesoplodonts, which may be more common in the Arabian Sea than the scarce literature suggests. If significant bycatch of beaked whales is confirmed, the massive tuna gillnet fishing effort in the Arabian Sea could have negative implications for their conservation status.
ARTICLE | doi:10.20944/preprints202109.0503.v1
Subject: Biology, Animal Sciences & Zoology Keywords: mitogenome; transmembrane proteins; substitution matrix; JTT matrix; molecular evolution; partitioned models; mixture models; RY coding; cyto-nuclear discordance
Online: 29 September 2021 (16:57:38 CEST)
Phylogenomic analyses have revolutionized the study of biodiversity, but they have revealed that estimated tree topologies can depend, at least in part, on the subset of the genome that is analyzed. For example, estimates of trees for avian orders differ if protein coding or non-coding data are analyzed. The bird tree is a good study system because the historical signal for relationships among orders is very weak, which should permit subtle non-historical signals to be identified, while monophyly of orders is strongly corroborated, allowing identification of strong non-historical signals. Hydrophobic amino acids in mitochondrially-encoded proteins, which are expected to be found in transmembrane helices, have been hypothesized to be associated with non-historical signals. We tested this hypothesis by comparing the evolution of transmembrane helices and extramembrane segments of mitochondrial proteins from 420 bird species, sampled from most avian orders. We estimated amino acids exchangeabilities for both structural environments and assessed the performance of phylogenetic analysis using each data type. We compared those relative exchangeabilities with values calculated using a substitution dataset for transmembrane helices from a variety of sampled set of nuclear- and mitochondrially-encoded proteins, allowing us to compare the bird-specific mitochondrial models with a general model of transmembrane protein evolution. To complement our amino acid analyses, we examined the impact of protein structure on patterns of nucleotide evolution. Models of transmembrane and extramembrane sequence evolution for amino acids and nucleotides exhibited striking differences, but there was no evidence for strong topological data type effects. However, incorporating protein structure into analyses of mitochondrially-encoded proteins improved model fit. Thus, we believe that considering protein structure will improve analyses of mitogenomic data, both in birds and in other taxa.
ARTICLE | doi:10.20944/preprints202109.0473.v1
Subject: Biology, Animal Sciences & Zoology Keywords: Cultural Use; Squirrel; gliding squirrel; tribes; Arunachal Pradesh
Online: 28 September 2021 (13:32:46 CEST)
Based on an ethnozoological study carried out from December 2018 to November 2019, this paper attempts to document the usage of squirrel species for various purposes by the Adi, Idu Mishmi, Miju Mishmi, Tangsa, Chakma and Monpa tribe living near the villages adjoining the protected areas located in the districts of East Siang, Lower Dibang Valley, Dibang Valley, Lohit, Changlang and West Kameng of Arunachal Pradesh, India. The paper also describes the use of certain squirrel species as a part of their traditional culture and lists their names in English, Vernacular and Latin names. The study has led to an understanding that eight squirrel species are used by the tribes for cultural, food and medicinal purposes. This work contributes to ethnozoological research by describing a knowledge system of squirrel species use and the association of squirrel species with the cultural believes of the tribes.
ARTICLE | doi:10.20944/preprints202109.0279.v1
Subject: Biology, Animal Sciences & Zoology Keywords: Primate cognition; scribbles; evolutive anthropology; art; aesthetics
Online: 16 September 2021 (11:20:42 CEST)
This study analyses 749 drawings of five female Bornean orang-utans (Pongo pygmaeus) at Tama Zoological Park in Japan. We searched for differences between individuals but also tried to identify possible temporal changes among the drawings of one individual, Molly, who drew almost 1,300 drawings from 2006 to 2016. An analysis of the drawings was carried out after collecting quantitative and qualitative variables. Our findings reveal evidence of differences in the drawing style of the five individuals as well as creative changes in Molly’s drawing style throughout her lifetime. Individuals differed in terms of the colours used, the space they filled but also the shapes (fan patterns, circles or loops) they drew. Molly drew less and less as she grew older, and we found a significant difference between drawings produced in winter, when orang-utans were kept inside and had less activity, and those produced during other seasons. Our results suggest that the drawing behaviour of these five orang-utans is not random and that differences among individuals might reflect differences of styles, states of mind but also motivation to draw.
Subject: Biology, Animal Sciences & Zoology Keywords: Amphipoda; invasive species; population genetics; miogenome; Ponto-Caspian; SNP
Online: 7 September 2021 (10:56:34 CEST)
The Ponto-Caspian region is the main donor of invasive amphipods to freshwater ecosystems, with at least 13 species successfully established in European inland waters. Dikerogammarus spp. and Pontogammarus robustoides are among the most successful, due to their strong invasive impact on local biota. However, genomic knowledge on these invaders is scarce, while phylogeography and population genetics have been based on short fragments of mitochondrial markers or nuclear mi-crosatellites. In this study, we provide: (i) reconstruction of four mitogenomes for four invasive gammarids; (ii) comparison between the structure of the newly obtained mitogenomes and to those from literature; (iii) SNP calling rates for individual D. villosus and D. haemobaphes from different invasion sites across Europe; and (iv) the first time-calibrated full mitogenome reconstruction of several Ponto-Caspian taxa. We have sequenced, assembled and annotated four mitogenomes, each from D. villosus, D. hae-mobaphes, D. bispinosus and Pontogammarus robustoides. We found that, in comparison to other gammarids, the mitogenomes of Ponto-Caspian species show a translocation between the tRNA-E and tRNA-R positions. Phylogenetic reconstruction using the mitogenomes identified that Ponto-Caspian gammarids form a well-supported group that originated in the Miocene. Our study supports paraphyly in the family Gammaridae. These mitogenomes serve as vital genetic resources for the development of new markers for PCR-based identification methods and demographic studies.
BRIEF REPORT | doi:10.20944/preprints202108.0407.v1
Subject: Biology, Animal Sciences & Zoology Keywords: Hair; HPA-axis; Koalas; Rehabilitation Sanctuary; Stress; Faeces; Glucocorticoids
Online: 20 August 2021 (08:52:07 CEST)
Koalas (Phascolarctos cinereus) are one of Australia's most charismatic native small marsupial species. Unfortunately, populations of koalas are rapidly declining throughout Australia and they continue to face increasing pressure from a changing ecosystem. Negative stimulants in the environment can elicit stress responses through activation of the hypothalamic-pituitary-adrenal (HPA) axis. Depending on the duration of the negative stimulant, the stress response can lead to either acute or chronic side effects, and is shown through the activation of the neuroendocrine stress system and the release of glucocorticoids (e.g., cortisol). Wild koalas entering clinical care face novel stressors that can be out of a wildlife carer's control. In this pilot study, we monitored physiological stress in three wild koalas at a wildlife rehabilitation centre in New South Wales, Australia. Acute and chronic stress was indexed non-invasively, with faecal samples taken to evaluate acute stress, and fur samples taken to evaluate chronic stress. Sampling occurred sporadically over four months, from the start of September 2018 to the end of December 2018. Results attempt to understand the stress response of koalas to negative stimulants in the environment by comparing faecal glucocorticoids on days where a known stressor was recorded with days where no known stressor was recorded. Furthermore, variations in faecal and fur glucocorticoids were compared between the three koalas in this study. To our knowledge, this is the first evidence of stress tracking of wild rescued koalas in a sanctuary. We suggest that further monitoring of baseline, acute and chronic stress will be needed to better understand how koalas respond to negative stimulants associated with clinical care.
ARTICLE | doi:10.20944/preprints202108.0323.v1
Subject: Biology, Animal Sciences & Zoology Keywords: racehorse; hoof; breakover; gallop; shoeing condition; surface; speed.
Online: 16 August 2021 (11:21:06 CEST)
Understanding the effect of horseshoe-surface combinations on hoof kinematics at gallop is relevant for optimising performance and minimising injury in racehorse-jockey dyads. This intervention study assessed hoof breakover duration in Thoroughbred ex-racehorses from the British Racing School galloping on turf and artificial tracks in four shoeing conditions: barefoot, aluminium-rubber composite (GluShu), aluminium and steel. Shoe-surface combinations were tested in a randomized order and horse-rider pairings (n=14) remained constant. High-speed video cameras (Sony DSC-RX100M5) filmed the hoof-ground interactions at 1000 frames per second. The time taken for a hoof marker wand fixed to the lateral hoof wall to rotate through an angle of 90 degrees during 384 breakover events was quantified using Tracker software. Data were collected for leading and non-leading front and hind limbs, at gallop speeds ranging from 23–56 km h-1. Linear mixed-models assessed whether speed, surface, shoeing condition or any interaction between these parameters (fixed factors) significantly affected breakover duration. Day and horse-rider pair were included as random factors and speed was included as a covariate. The significance threshold was set at p<0.05. For all limbs, breakover times decreased as gallop speed increased (p<0.0005), although a greater relative reduction in breakover duration for hindlimbs was apparent beyond approximately 45 km h-1. Breakover duration was longer on turf compared to the artificial surface (p≤0.04). In the non-leading hindlimb only, breakover duration was affected by shoeing condition (p=0.025) and an interaction between shoeing condition and speed (p=0.023). Future work seeks to relate these results to hoof accelerometer data.
REVIEW | doi:10.20944/preprints202108.0315.v1
Subject: Biology, Animal Sciences & Zoology Keywords: amylase; carbohydrates; crustaceans; decapods; digestion; feeding habits; lobster.
Online: 16 August 2021 (09:33:48 CEST)
Decapod crustaceans are a very diverse group and have evolved to suit a wide variety of diets. Alpha-amylases enzymes, responsible for starch and glycogen digestion, have been more thoroughly studies in herbivore and omnivore than in carnivorous species. We used information on the α-amylase of a carnivorous lobster as a connecting thread to provide a more comprehensive view of α-amylases across decapods crustaceans. Omnivorous crustaceans such as shrimps, crabs and crayfish present relatively high amylase activity respect to carnivorous. Yet, contradictory results have been obtained and relatively high activity in some carnivores has been suggested to be a remnant trait from ancestor species. Here we provided information sustaining that high enzyme sequence and overall architecture conservation do not allow high changes in activity, and that differences among species may be more related to number of genes and isoforms, as well as transcriptional and secretion regulation. However, recent evolutionary analyses revealed that positive selection might have also occurred among distant lineages with feeding habits as a selection force. Some biochemical features of decapod α-amylases can be related with habitat or gut conditions, while less clear patterns are observed for other enzyme properties. Likewise, while molt cycle variations in α-amylase activity are rather similar among species, clear relationships between activity and diet shifts through development cannot be always observed. Regarding the adaptation of α-amylase to diet, juveniles seem to exhibit more flexibility than larvae, and it has been described variation in α-amylase activity or number of isoforms due to the source of carbohydrate and its level in diets, especially in omnivore species. In the carnivorous lobster, however, no influence of the type of carbohydrate could be observed. Also, lobsters were not able to fine-regulate α-amylase gene expression in spite of large changes in carbohydrate content of diet, while retaining some capacity to adapt α-amylase activity to very low carbohydrate content in the diets. In this review, we raised arguments for the need of more studies on the α-amylases of less studied decapods groups, including carnivorous species which rely more on dietary protein and lipids, to broad our view of α-amylase in decapods crustaceans.
ARTICLE | doi:10.20944/preprints202108.0198.v1
Subject: Biology, Animal Sciences & Zoology Keywords: depolymerizing enzyme; debranching enzyme; xylanase; arabinofuranosidase; feruloyl esterase; arabinoxylan; digestion; growth performance; volatile fatty acid; piglet
Online: 9 August 2021 (14:56:56 CEST)
This study was aimed to investigate the effect of xylan depolymerizing enzyme namely endo-xylanase (Xyn) combined with debranching enzymes namely arabinofuranosidase (Afd) and feruloyl esterase (FE) on digestion, growth performance and intestinal volatile fatty acid profile of piglets. The in vitro experiments were firstly conducted to examine the enzymological properties of Xyn, Afd and FE, the synergy among these enzymes, together with the effect of combination of these enzymes on digestion of piglet diet. The in vivo experiment was then implemented by allocating 270 35-d-old postweaning piglets into 3 treatment groups: control group, Xyn group and (Xyn+Afd+FE) group. Each group had 6 replicates (15 piglets/replicate). The results revealed a satisfying thermostability and pH stability of Xyn, Afd and FE. Combination of Xyn, Afd and FE had a superiority (P < 0.05) over Xyn alone and its combination with Afd or FE in promoting degradation of different bran fibers rich in arabinoxylan (Abx). Treatment with combination of Xyn, Afd and FE had advantages over Xyn alone to induce increasing trends (P < 0.10) of in vitro digestibility of dietary nutrients (dry matter, crude protein, crude ash and gross energy) and piglet growth performance (average daily gain, final body weight and feed efficiency), concurrent with a reduction (P < 0.05) of diarrhea rate and increases (P < 0.05) in cecal acetic acid, butyric acid and total volatile fatty acids concentrations as well as pH value of piglets. Collectively, combination of Xyn, Afd and FE was efficient in benefiting degradation of Abx in brans, as well as improving digestion, growth performance and intestinal volatile fatty acid profile of piglets.
ARTICLE | doi:10.20944/preprints202107.0266.v1
Subject: Biology, Animal Sciences & Zoology Keywords: fullerene; nanomaterials; clownfish; metabolomic; muscle
Online: 12 July 2021 (14:06:45 CEST)
Fullerene nanomaterials exposure often causes a variety of diseases. Many studies have pointed out that fullerene nanomaterials can be studied in fish. However, there are few studies on health risk assessment of clownfish with lower doses of fullerene nanoparticles or different exposure durations. In this study, we set 1.5% and 3.0% for low- and high-dose fullerene nanomaterials exposure concentrations, respectively. Meanwhile, we performed a time-series analysis to investigate that the activation of lipid and amino acids metabolism after fullerene nanomaterials exposure in clownfish. 1368 metabolites were detected from clownfish by using liquid chromatography-mass spectrometry (LC-MS) analyses. Our results suggest that exposure to lower fullerenes nanoparticles may have a certain promoting effect on clownfish overall length, body length and weight. At the same time, the activation of potential metabolic pathways enriched by different metabolites in KEGG pathway may also indicate the positive promoting effect of fullerene nanoparticles after exposure. The present work indicates that it is particularly important to find the concentration window for fullerene nanomaterials to improve government safety guidelines, especially when these are applied to assess the health risk of human.
CONCEPT PAPER | doi:10.20944/preprints202106.0578.v1
Subject: Biology, Animal Sciences & Zoology Keywords: homology; developmental mechanism; evidential integration; eumetazoan body plan; phylogenetics
Online: 23 June 2021 (11:45:06 CEST)
Reconstructing ancestral species is a challenging endeavour: fossils are often scarce or enigmatic, and inferring ancestral characters based on novel molecular approaches (e.g. comparative genomics or developmental genetics) has long been controversial. A key philosophical challenge pertinent at present is the lack of a theoretical framework capable of evaluating inferences of homology made through integration of multiple kinds of evidence (e.g. molecular, developmental, or morphological). Here, I present just such a framework. I start with a brief history and critical assessment of attempts at inferring morphological homology through developmental genetics. I then bring attention to a recent model of homology, namely Character Identity Mechanisms (DiFrisco, Love, & Wagner, 2020), intended partly to elucidate the relationships between morphological characters, developmental genetics, and homology. I utilise and build on this model to construct the evaluative framework mentioned above, which judges the epistemic value of evidence of each kind in each particular case based on three proposed criteria: effectiveness, admissibility, and informativity, as well as providing a generalised guideline on how it can be scientifically operationalised. I then point out the evolution of the eumetazoan body plan as a case in point where the application of this framework can yield satisfactory results, both empirically and conceptually. I will conclude with a discussion on some potential implications for more general philosophy of biology and philosophy of science, especially surrounding evidential integration, models and explanation, and reductionism.
ARTICLE | doi:10.20944/preprints202106.0279.v1
Subject: Biology, Animal Sciences & Zoology Keywords: Blood-Brain Barrier, Penetrating Peptides, Machine learning techniques, Drug delivery, Therapeutic peptides
Online: 10 June 2021 (08:13:15 CEST)
Blood-brain-barrier is a major obstacle in treating brain-related disorders as it does not allow to deliver drugs in the brain. In order to facilitate delivery of drugs in brain, we developed a method for predicting blood-brain-barrier penetrating peptides. These blood-brain barriers penetrating peptides (B3PPs) can act as therapeutic as well as drug delivery agents. We trained, tested, and evaluated our models on blood-brain-barrier peptides obtained from the B3Pdb database. First, we compute a wide range of peptide features then we select relevant peptide features. Finally, we developed numerous machine learning-based models for predicting blood-brain-barrier peptides using selected features. Our model based on random forest performed best on the top 80 selected features and achieved a maximum 85.08% accuracy with 0.93 AUROC. We also developed a web server, B3pred that implements our best models. It has three major modules that allow users to; i) predict B3PPs, ii) scanning B3PPs in a protein sequence, and iii) designing B3PPs using analogs. Our web server and standalone software is freely available at https://webs.iiitd.edu.in/raghava/b3pred/.
Subject: Biology, Animal Sciences & Zoology Keywords: convolutional neural networks; horse emotion recognition; horse emotion
Online: 7 June 2021 (12:42:05 CEST)
Creating intelligent systems capable of recognizing emotions is a difficult task, especially when looking at emotions in animals. This paper describes the process of designing a “proof of concept” system to recognize emotions in horses. This system is formed by two elements, a detector and a model. The detector is a fast region-based convolutional neural network that detects horses in an image. The model is a convolutional neural network that predicts the emotions of those horses. These two elements were trained with multiple images of horses until they achieved high accuracy in their tasks. 400 images of horses were collected and labeled to train both the detector and the model while 80 were used to validate the system. Once the two components were validated, they were combined into a testable system that would detect equine emotions based on established behavioral ethograms indicating emotional affect through head, neck, ear, muzzle and eye position. The system showed an accuracy of between 69% and 74% on the validation set, demonstrating that it is possible to predict emotions in animals using autonomous intelligent systems. Such a system has multiple applications including further studies in the growing field of animal emotions as well as in the veterinary field to determine the physical welfare of horses or other livestock.
CONCEPT PAPER | doi:10.20944/preprints202105.0767.v1
Subject: Biology, Animal Sciences & Zoology Keywords: evidential integration; causal explanation; early animal evolution; phylogenetics; macroevolution; evolutionary scenario; cross-disciplinary research
Online: 31 May 2021 (12:25:44 CEST)
Molecular methods have revolutionised virtually every area of biology, and metazoan phylogenetics is no exception: molecular phylogenies, molecular clocks, comparative phylogenomics, and developmental genetics have collectively transformed our understanding of the evolutionary history of animals. Moreover, the diversity of methods and models within molecular phylogenetics has resulted in significant disagreement among molecular phylogenies as well as between these and traditional phylogenies. Here, I argue that tackling this multifaceted problem lies in integrating evidence to infer the best evolutionary scenario. I begin with an overview of recent developments in early metazoan phylogenetics, followed by a discussion of key conceptual issues in phylogenetics revolving around phylogenetic evidence and theory. I then argue that integration of different kinds of evidence is necessary for arriving at the best evolutionary scenario rather than the best-fitting cladogram. Finally, I discuss the prospects of this view in stimulating interdisciplinary cross-talk in early metazoan research and beyond.
ARTICLE | doi:10.20944/preprints202103.0409.v1
Subject: Biology, Animal Sciences & Zoology Keywords: coronavirus; horseshoe bats; reservoir hosts; Indochina; China; herd immunity.
Online: 16 March 2021 (09:39:15 CET)
To date, viruses closely related to SARS-CoV-2 have been reported in four bat species: Rhinolophus acuminatus, Rhinolophus affinis, Rhinolophus malayanus, and Rhinolophus shameli. Here, we analysed 343 sequences of the mitochondrial cytochrome c oxidase subunit 1 gene (CO1) from georeferenced bats of the four Rhinolophus species identified as reservoirs of SARS-CoV-2-like viruses. Haplotype networks were constructed in order to investigate patterns of genetic diversity among bat populations of Southeast Asia. No strong geographic structure was found for the four Rhinolophus species, suggesting high dispersal capacity. The ecological niche of SARS-CoV-2 like viruses was predicted using the four localities of bat SARS-CoV-2-like viruses and the localities where bats showed identical or very similar CO1 haplotypes than virus-positive bats. The ecological niche of SARS-CoV-like viruses was deduced from the localities where bat SARS-CoV-like viruses were previously detected. The results show that the ecological niche of SARS-CoV2-like viruses includes several regions of mainland Southeast Asia whereas that of SARS-CoV-like viruses is mainly restricted to China. In agreement with these results, human populations in Laos, Vietnam, Cambodia, and Thailand appear to be much less affected by the Covid-19 pandemic than other countries of Southeast Asia. In the climatic transitional zone between the two ecological niches (southern Yunnan, northern Laos, northern Vietnam, and possibly Hainan and Taiwan), genomic recombination between highly divergent viruses is more likely to occur. Since recombinant viruses can threaten the benefit of vaccination campaigns, these regions should be under surveillance.
ARTICLE | doi:10.20944/preprints202102.0227.v1
Subject: Biology, Animal Sciences & Zoology Keywords: rehabilitation; stress; reptiles; injury; disease; euthanasia; trauma; clinical care
Online: 9 February 2021 (09:23:18 CET)
Direct and indirect anthropogenic factors play a massive role in driving wildlife species towards extinction. Longitudinal retrospective studies identify key ‘factors’ responsible for the decline in numbers of wildlife, however, lack the reasoning behind the events leading to mortality. The overarching aim of this study was to categorize these ‘factors’ into different stressor categories faced by reptiles to understand its impact on an individual, and to compare how each stressor category influences the survival of an individual. The results from this study indicated that almost half of the number of reptiles being hospitalized were due to exposure to preliminary stressors such as lawn mowing incidents and pet attack. Primary and secondary admissions were fairly equal in number, however the mortality rate for secondary admissions was drastically high (~80%). The discussion integrates species’ ecology and stress physiology which can prove to have multi-faceted benefits across the fields of ecology and animal welfare. Ecologists can use the results from this study to comprehend species’ activity patterns to better plan reptilian conservation programs, whereas, for wildlife clinicians and rehabilitators, assignment of stressor categories could be a beneficial tool for bolstering the welfare monitoring program for small native reptiles in clinical settings.
Subject: Biology, Animal Sciences & Zoology Keywords: intramuscular fat; prediction; image analysis; Bísaro pork
Online: 13 January 2021 (13:16:19 CET)
This work presents an analytical methodology to predict meat juiciness (discriminant semi-quantitative analysis using groups of intervals of intramuscular fat) and intramuscular fat (regression analysis) in Longissimus thoracis et lumborum (LTL) muscle of Bísaro pigs using as independent variables the animal carcass weight and parameters from color and image analysis. These are non-invasive and non-destructive techniques which allow development of rapid, easy and inexpensive methodologies to evaluate pork meat quality in a slaughterhouse. The proposed predictive supervised multivariate models were non-linear. Discriminant mixture analysis to evaluate meat juiciness by classified samples into three groups—0.6 to 1.1%; 1.25 to 1.5%; and, greater than 1.5%. The obtained model allowed 100% of correct classifications (92% in cross-validation with seven-folds with five repetitions). Polynomial support vector machine regression to determine the intramuscular fat presented R2 and RMSE values of 0.88 and 0.12, respectively in cross-validation with seven-folds with five repetitions. This quantitative model (model’s polynomial kernel optimized to degree of three with a scale factor of 0.1 and a cost value of one) presented R2 and RSE values of 0.999 and 0.04, respectively. The overall predictive results demonstrated the relevance of photographic image and color measurements of the muscle to evaluate the intramuscular fat, rarther than the usual time-consuming and expensive chemical analysis.
Subject: Biology, Animal Sciences & Zoology Keywords: Stress; fear; anxiety; aggression; veterinary visit; low-stress handling; counterconditioning; behaviour modification; anxiolytic medication; psychoactive drugs; dogs; cats
Online: 5 January 2021 (11:15:02 CET)
A high proportion of dogs and cats are fearful during veterinary visits, which in some cases may escalate into aggression. Here, we discuss factors that contribute to negative emotions in a veterinary setting and how these can be addressed. We briefly summarise the available evidence for the interventions discussed. The set-up of the waiting area (e.g. spatial dividers; elevated places for cat carriers), tailoring the examination and the treatment to the individual, considerate handling (minimal restraint when possible, avoiding leaning over or cornering animals) and offering high-value food or toys throughout the visit can promote security and, ideally, positive associations. Desensitisation and counterconditioning are highly recommended both to prevent and address existing negative emotions. Short-term pain from injections can be minimised by using tactile and cognitive distractions and topical analgesics, which are also indicated for painful procedures such as ear cleanings. Recommendations for handling fearful animals to minimise aggressive responses are discussed. However, anxiolytics or sedation should be used whenever there is a risk of traumatising an animal or for safety reasons. Stress-reducing measures can decrease fear and stress in patients and consequently their owners, thus strengthening the relationship with the clients as well as increasing the professional satisfaction of veterinary staff.
REVIEW | doi:10.20944/preprints202012.0283.v2
Subject: Biology, Animal Sciences & Zoology Keywords: SARS-CoV-2; Covid-19; wildlife; host-switching; reservoirs; risk assessment; surveillance
Online: 24 December 2020 (09:20:20 CET)
The novel coronavirus SARS-CoV-2 likely emerged from a wildlife source with transmission to humans followed by rapid geographic spread throughout the globe and dramatic impacts on both human health and global economies. Since the onset of the pandemic, there have been several instances of human-to-animal transmission involving companion, farmed and zoo animals, and one instance of infection in a wild mink, with the clear potential for further spread into free-living wildlife. The establishment of reservoirs of infection in wild animals would create significant challenges to infection control in humans and could pose a threat to the welfare and conservation status of wildlife. Herein, we discuss the potential for exposure, maintenance and onward transmission of SARS-CoV-2 in an initial selection of wild and feral species (bats, canids, felids, mustelids, great apes). Targeted surveillance and dynamic risk assessment are important tools for the early detection of infection in wildlife and a means of collating and synthesising emerging information in a rapidly changing situation. Such efforts should be integrated with public health information to provide insights into the potential role of wild mammals in the continuing epidemiology of SARS-CoV-2. This approach should also be adopted to address the wider need to proactively assess threats to human and animal health from other diseases that may emerge from wildlife.
Subject: Biology, Animal Sciences & Zoology Keywords: CTs molecular weight; ruminal fermentation; bio-hydrogenation; milk compositions; goat
Online: 20 September 2020 (14:44:56 CEST)
The aim of this study was to investigate the effect of condensed tannin (CTs) with differing molecular weight on their capacity to modify the fatty acid profile in milk. Twenty multiparous crossbred lactating dairy goats were assigned in a randomized complete block design (RCBD), and were subjected to receive the dietary treatments as followings; T1: control (with no CTs supplementation), T2: supplemented with mangosteen peel in a concentrate as a source of low molecular weight CTs at level of 3.0 %DM of CTs equivalent, T3: supplemented with the same diet with T2 but added with polyethylene glycol (PEG, as tannin inactivator) as the control of T2, and T4: supplemented with quebracho CT extract (UNITAN ATO, Buenos Aires, Argentina; 75-77 % tannins) in a concentrate as a source of high molecular weight CTs at level of 3.0 %DM of CTs equivalent, and T5) supplemented with the same diet with T4 but added with PEG as the control of T4. No significant change was detected for feed intake and nutrient digestibility indicate that CTs at level of 3.0 %DM of diet did not showed the detrimental effect to feed intake and nutrient digestibility, however, ruminal fermentation parameters and milk yield and milk compositions did not affected by different source of CT inclusion.
REVIEW | doi:10.20944/preprints202009.0396.v1
Subject: Biology, Animal Sciences & Zoology Keywords: Feeding behavior; Feeding habits; Physiology; Hexacorallia; Octocorallia; Ceriantharia
Online: 17 September 2020 (10:48:18 CEST)
In this study, we performed a bibliographical review examining the scientific literature on “feeding in Anthozoa” theme for the period from 1890 to 2019, using scientific databases (Google Scholar) supplemented with additional literature. This study categorized published scientific papers on this topic by decade of publication, target taxa, variability of species studied in each order and main themes studied. As a result, 153 studies were found, and based on their content, it was observed that within Anthozoa, there has been a concentration of feeding studies on species in the orders Actiniaria (Hexacorallia), Scleractinia (Hexacorallia), and Alcyonacea (Octocorallia). This indicates that the other remaining orders of the group have been comparatively neglected with regards to their feeding aspects. Therefore, as data on feeding in some groups of Anthozoa are scarce, studies need to be carried out to fill the gaps that permeate this important benthic group, in order to better understand their ecology.
ARTICLE | doi:10.20944/preprints202009.0270.v1
Subject: Biology, Animal Sciences & Zoology Keywords: caprine arthritis-encephalitis virus; goat; antigens; antibodies; proviral DNA
Online: 12 September 2020 (09:57:24 CEST)
In this study we have approached the detection of caprine arthritis-encephalitis virus (CAEV) using a multi target approach testing with both ELISA and an in-house real-time PCR test to investigate the prevalence of CAEV in goats from hobbyist farms in the Republic of Tatarstan, Russia. Animals from three hobbyist farms were used in this study. The animals from two farms (n=13 for F1 and n=8 for F2) had clinical signs of arthritis and mastitis. In the third farm (n=15 for F3), all goats were homebred and had no contact with imported animals. CAEV antibodies (ELISA targets TM ENV and GaG genes) were detected in serum samples from two farms (F1 and F2), indicating a seroprevalence 87.50-92.31%. Specific CAEV antibodies were also detected in milk samples. CAEV proviral DNA was detected in 53.85-62.50%. Results from all tests performed in the third farm (F3) were negative, indicting all tests apparent specificity of 100%. The results of this work show that CAEV is circulating and present in small hobbyist goat farms in Russia. Serological and molecular tests could be of importance for CAEV control and eradication programs in Russia for hobbyist goat farms.
ARTICLE | doi:10.20944/preprints202009.0261.v1
Subject: Biology, Animal Sciences & Zoology Keywords: Arachnid; Scorpion; Spider; Pet; Bite; Sting; Envenomation; Pain; Symptoms
Online: 12 September 2020 (03:40:40 CEST)
With a global estimate of tens of thousands of arachnid enthusiasts, spiders and scorpions are gaining increasing popularity as pets in industrialised countries in Europe, Northern America and Asia. As most spiders and all scorpions are venomous and due to their mostly negative image in the public media, several governments are already considering introducing legislation to regulate the domestic care of potentially dangerous captive animals. We aimed to investigate the circumstances and effects of exposure to arachnids kept in captivity. Thus, we collected and analysed data from 335 reported bites and stings attributed to pet arachnids. Our data revealed that on average there were less than 20 recorded envenomations per year with ~90% preventable by due care. We also categorized the severity of the resulting symptoms and found that the vast majority of symptoms were either local (60.9%) or moderate (33.4%), 4.8% were asymptomatic, only 0.9% were severe and no fatalities were recorded. Based on our database of bite and sting reports, we performed a risk assessment for arachnid pet ownership and concluded that with the proper care arachnids can be safely kept as pets and pose a lower risk than many other recreational activities.
Subject: Biology, Animal Sciences & Zoology Keywords: Cherax quadricarinatus; red claw; substrate; survival; growth; molting
Online: 7 September 2020 (09:53:47 CEST)
The experiment was conducted to determine the effects of different substrate on the survival, growth and total number of moulting in juvenile red claw, Cherax quadricarinatus for aquaculture purposes. In the present study, there is a problem in culturing juvenile stage of C. quadricarinatus especially for survival and growth. Thus, a substrate was used to improve the survival and growth of C. quadricarinatus especially for the intensive system due C. quadricarinatus need a large space to survive. C. quadricarinatus with initial body weight from 1.10 to 2.90 g, total length 2.82 to 4.36 cm were placed in tanks (80 L in volume capacity) with 55L in water and there are eight juveniles in each tank. Treatment tanks were introduced with coral as treatment 2 and pipe as treatment 3. While, tank without shelters was set as control (treatment 1). Black nets were installed on top of each for circumventing foreign objects enter the tank and protect from direct sunlight. Besides, C. quadricarinatus were acclimatized for seven days before started the experiment. The experiment was done in duplicate and conducted for 60 days and C. quadricarinatus were fed twice daily (morning and evening) based on 5% of body weight. From the total of two replications, C. quadricarinatus for treatment 2 (coral) were significantly in survival (81.25±8.84%), weight gain (347.36 ±6.04%), specific growth rate (2.50±0.02%), carapace length (32.93 ± 0.93 %) and total number of moulting (55.00±2.93%) compared to other treatments. Besides, the control treatment (treatment 1) has a lower percentage in survival (37.50±8.84%) weight gain (122.60±20.51%) specific growth rate (1.33±0.15%), carapace length (25.84±0.33 %) and total number of molting (29.00±0.71%). As a conclusion, usage of coral as the substrate in the rearing tank showed improvement in survival, weight gain, specific growth rate, carapace length and total number of molting in C. quadricarinatus. The coral can use in rearing tank for increase the growth and survival for a small scale and not for commercial. In addition, in the environmental aspect, the present study showed the benefit of replacing the use of PVC pipes with the natural structure of dead coral.
REVIEW | doi:10.20944/preprints202009.0101.v1
Subject: Biology, Animal Sciences & Zoology Keywords: facial expressions; pain; grimace scales; mice; rat; rabbit
Online: 4 September 2020 (11:18:42 CEST)
Animals’ facial expressions have been widely used as a readout for emotion. Scientific interest in the facial expressions of laboratory animals has centered primarily on negative experiences, such as pain, experienced as a result of scientific research procedures. Recent attempts to standardize evaluation of facial expressions associated with pain in laboratory animals has culminated in the development of “grimace scales”. In the context of laboratory animals, these have been developed and evaluated for mice, rats, rabbits, sheep, and ferrets. The prevention or relief of pain in laboratory animals is a fundamental requirement for in vivo research to satisfy community expectations. However, to date it appears that the grimace scales have not seen widespread implementation as clinical pain assessment techniques in biomedical research. In this review, we discuss some of the barriers to implementation of the scales in clinical laboratory animal medicine, progress made in automation of collection, and suggest avenues for future research.
Subject: Biology, Animal Sciences & Zoology Keywords: animal suffering; animal welfare; fires; wild animals
Online: 28 August 2020 (08:50:40 CEST)
Animals living in the wild are exposed to numerous challenges, such as fires, that can lead to animal suffering. The impacts of fire have been studied in different branches of ecology, but studies of its effects on the welfare of individual animals remain scarce. The current review aims to synthesize a sample of relevant aspects regarding fire’s negative effects on wild animals. This review provides a better understanding of how fire compromises animal welfare, providing an example of how to use the knowledge gathered in ecology studies to examine the welfare of wild animals. It can help raise concern for the situation of wild animals as individuals, and to develop the field of welfare biology, by identifying promising future lines of research. The fundamentals of carrying out future work to design protocols for rescuing animals or preventing the harms they can suffer in fires is also explored.
ARTICLE | doi:10.20944/preprints202008.0518.v1
Subject: Biology, Animal Sciences & Zoology Keywords: Cattle; RNF128; RNF34; Single nucleotide polymorphisms
Online: 24 August 2020 (09:44:34 CEST)
An experiment was conducted to investigate the role of single nucleotide polymorphisms of the gene RNF34 3′UTR-588 G>A and RNF128 I1-2380C>T with carcass and meat quality traits of Chinese Simmental-cross steers. METHOD: We performed sequencing and restriction enzyme digestion to detect genotypes of RNF34 3′UTR-588 G>A and RNF128 I1-2380C>T. Then, we analyzed the association of novel single nucleotide polymorphisms in intron regions of the RNF128 gene and in the 3′UTR region of RNF34 and meat quality traits of Chinese Simmental-cross steers. RESULTS: Statistical analyses revealed that SNP of RNF128 (I1-2380C>T) was significantly associated with dressed weight, forepaw weight, carcass depth, carcass brisket depth, hind legs length (P<0.05), etc. And RNF34 (3′UTR-588 G>A) were significantly associated with testis weight, kidney weight, tare weight (P<0.05), etc. CONCLUSION: Our findings suggest that polymorphisms in RNF34 and RNF128 might be important genetic factors that influence carcass and meat quality in beef cattle. Thus, they might be useful markers for meat quality traits in future marker-assisted selection programs in beef cattle breeding and production.
COMMUNICATION | doi:10.20944/preprints202008.0465.v1
Subject: Biology, Animal Sciences & Zoology Keywords: Red ants; Ex-situ; Conservation; Hatchery; Sea turtles; Lepidochelys olivacea
Online: 20 August 2020 (13:20:44 CEST)
Abstract: Predation of eggs and emerging hatchlings of olive ridley turtle (Lepidochelys olivacea) by wild animals and feral dogs are known. They reduce the hatching success rate considerably affecting the conservation management of this vulnerable species. Hatchery management is practised in India to overcome predation. Ant predation is a serious threat to turtle nest protected by ex situ or in situ erected hatchery. This article reports the first direct evidence of turtle eggs predation by Dorylus orientalis Westwood, 1835 commonly called red ants. Native to India, Oriental, Indo Australian and Palearctic regions they are notorious as an agricultural pest. Chlorpyrifos pesticides recommended for their control could become fatal for the developing embryos of turtles if applied near the hatchery. In the turtle nesting site of the west coast of India, D. orientalis has more of an ecological role than as a pest. Natural pesticide such as Neem powder (Azadirachta indica) shows promising results for preventing their infestation.
HYPOTHESIS | doi:10.20944/preprints202008.0459.v1
Subject: Biology, Animal Sciences & Zoology Keywords: colonial life-history; major evolutionary transition (MET); cooperation; modularity; biological information; heterochrony; tunicate
Online: 20 August 2020 (13:05:33 CEST)
The diversification of life-histories is mediated by cooperation, innovations of biological information, modularity, and heterochrony in developmental processes. These processes are defined, contextualized, and exemplified, studying the evolution of coloniality (i.e. life-history involving modularization of the multicellular individual) in the family of benthic tunicates Styelidae. This study proposes that in these colonial tunicates there is an inter-generational division of labor, where one generation is feeding, a second is developing by morphogenetic processes, and a third is aging by programmed cell death and phagocytosis. The communication system developed in these colonies is mediated, by changes in proportion, location, and gene expression of specialized blood cells. Colonial life-history in animals is related to the reduction of individual size; development of extra-corporeal tissues to interconnect zooids; the inter-generational division of labor; and the reduction of zooid’s individuality. Processes analogous with the widely accepted major evolutionary transitions (METs), suggesting that coloniality could be studied as a MET. The understanding of colonial life-histories could provide information about key mechanisms for life diversification.
ARTICLE | doi:10.20944/preprints202008.0395.v1
Subject: Biology, Animal Sciences & Zoology Keywords: economic analysis; conservation; P.monodon; by-catch; livelihood; seed collector; post larvae collector; coastal aquaculture; Indian Sundarbans
Online: 19 August 2020 (02:53:11 CEST)
The livelihood of most of the fishers in Indian Sundarbans is dependent on Penaeus monodon post larvae fishing. These post larvae collectors are socially backward lacking economic security. The activity of collecting Penaeus monodon post larvae for rearing in aquaculture, destroy other aquatic species. Many other juveniles of shellfish and fin fish were destroyed in the process of collection of Penaeus monodon post larvae. The removal of juveniles before they reach maturity disturbs the ecological chain by hampering breeding processes and may cause extinction of some fish species in the long run. The present study is an attempt to estimate the economic value of juveniles destroyed in the collection of (Penaeus monodon) post larvae. In total 32 species were identified in P.monodon the post larval by-catch. The economic loss is assessed based on estimating biomass by taking a length-weight relationship from published literature. Further, the paper illustrates how does a profit enterprise is linked with natural resource exploitation. The paper explores government policy and nature conservation issues for social justice and effective conservation.In conclusion, suggestions are given to reduce the burden of livelihood on natural resources to the extent of exploitation and to strengthen institution and policy-making considering socio-ecological vulnerabilities of the area.
ARTICLE | doi:10.20944/preprints202008.0354.v1
Subject: Biology, Animal Sciences & Zoology Keywords: seahorse; effective day degrees; temperature; stable isotopes; Hippocampus
Online: 17 August 2020 (08:11:00 CEST)
Relations between nutrient assimilation and growth rate in fishes may vary with abiotic factors such as temperature. The effects of feeding status, ontogeny and temperature regimes (15, 18 and 21 °C) on stable isotope (δ13C and δ15N) patterns were assayed and modelled in juveniles of the seahorse Hippocampus guttulatus. The use of effective day degrees (D°eff) and chronological time (age) were compared as development progress indices. Newborn seahorses were maintained at three temperature levels both deprived of food (5 days) or fed (30 days) on copepods or/and Artemia. Isotopic signatures in fed seahorses clearly differed from those in unfed juveniles. Temperature had a significant effect on δ13C values in fed juveniles throughout the experimental period. δ15N values also varied significantly with age, but not with temperature level. Faster growth and food assimilation in seahorses held at 18 and 21 °C were supported by faster variations in isotopic values. Our findings demonstrate that effective day degrees should be preferred over chronological time as index of developmental progress in temperature fluctuating scenarios or for comparative studies.
REVIEW | doi:10.20944/preprints202008.0313.v1
Subject: Biology, Animal Sciences & Zoology Keywords: drug resistance; natural diversity; C. elegans; anthelmintics
Online: 14 August 2020 (08:56:53 CEST)
Parasitic nematode infections impact human and animal health globally, especially in the developing world. Anthelmintic drugs are the major line of defense against these infections, but the arsenal is limited. Additionally, anthelmintic resistance is widespread in veterinary parasites and an emerging threat in human parasites. Discoveries of the mode of action of these drugs and mechanisms of resistance have predominantly come from studies of a related non-parasitic nematode species, Caenorhabditis elegans, and the parasitic nematode Haemonchus contortus. Here, we discuss recent progress understanding anthelmintic resistance using these two species and how that progress relates to laboratory and field-based studies of veterinary helminths. We present a powerful approach enabled by the strengths of both nematode species to understand mechanisms of resistance and modes of action of anthelmintic drugs.
ARTICLE | doi:10.20944/preprints202008.0276.v1
Subject: Biology, Animal Sciences & Zoology Keywords: Free-range; backfat layers; intramuscular fat; Iberian pig; subcutaneous fat.
Online: 12 August 2020 (08:11:09 CEST)
Twenty-four extensively-reared Iberian pigs were used to study the influence of fattening period length (30, 60 or 90 days) on the fatty acid profiles of intramuscular and subcutaneous fat and the relationships between both profiles. Regarding fatty acid (FA) percentage, PUFA was greater in backfat and MUFA was greater in intramuscular fat (IMF), regardless fattening period length. The longer fattening period increased MUFA content in backfat (which had a more marked change in oleic acid) and decreased PUFA content in backfat and IMF, but it did not affect SFA content. Within the 3-layers subcutaneous backfat, SFA content was greater in the inner layer, MUFA was greater in the outer layer and PUFA was greater in both of these layers. The few differences in FA composition between both adipose tissues suggest that the changes due to the feeding regime are slow and, therefore, although the length of the fattening phase was increased, the fatty acid profile did not change substantially. The strong relationship between the FA profiles of IMF and backfat might be used to predict one profile from the other one when this latter was more readily available for sampling or analytical reasons.
ARTICLE | doi:10.20944/preprints202008.0214.v1
Subject: Biology, Animal Sciences & Zoology Keywords: stable isotopes; preservation; syngnathids; seahorses; pipefishes; conversion models
Online: 8 August 2020 (18:13:47 CEST)
Isotopic stable analysis (SIA) is a powerful tool in the assessment of different types of ecological and physiological studies. For that, different preservative methods for the samples are commonly used prior to isotopic analysis. The effects of various preservative methods (drying, freezing, ethanol and formaldehyde) have been analyzed for C:N ratio, δ13C and δ15N on a variety of tissues including dorsal fins (three seahorse and two pipefish species), seahorse newborns (three seahorses species), and prey (copepods and different stages of Artemia) commonly used to feed the fishes in rearing conditions. The aims of the study were to: (i) evaluate isotopic effects of preservation methods across tissues; and (ii) construct the first conversion models available in syngnathid fishes. The preservation in ethanol and to a lesser extend in formaldehyde significantly affected δ13C values, whereas δ15N signatures were not affected significantly. Due to their low lipid content, the isotopic signals in fish fins were almost unaffected, supporting the suitability of dorsal fins as a convenient tool in isotopic studies on vulnerable species such as syngnathids. The regression equations provided permit the successful conversion of δ13C and δ15N values between preservative treatments. The conversion models can be applied to isotopic studies in the field and in the laboratory.
ARTICLE | doi:10.20944/preprints202008.0103.v1
Subject: Biology, Animal Sciences & Zoology Keywords: chicken; Newcastle disease; spleen; immune response; gene expression; RNA-seq
Online: 4 August 2020 (16:09:52 CEST)
As a major infectious disease in chickens, Newcastle disease causes considerable economic losses in the poultry industry, especially in developing countries where there is limited access to effective vaccination. Therefore, enhancing resistance to the virus in commercial chickens through breeding is a promising way to promote poultry production. In this study, we investigated gene expression changes at 2 and 6 dpi after infection at day21 with a lentogenic Newcastle disease virus in a commercial egg-laying chicken hybrid using RNA sequencing analysis. By comparing NDV challenged and nonchallenged groups, 526 differentially expressed genes (DEGs) (FDR < 0.05) were identified at 2 dpi, and only 36 at 6 dpi. For the DEGs at 2 dpi, IPA analysis predicted inhibition of multiple signaling pathways in response to NDV that regulate immune cell development and activity, neurogenesis and angiogenesis. Upregulation of Interferon Induced Protein with Tetratricopeptide Repeats 5 (IFIT5) in response to NDV was consistent between the current and most previous studies. Sprouty RTK Signaling Antagonist 1 (SPRY1), a DEG in the current study is located in a significant QTL associated with virus load at 6 dpi in the same population. These identified pathways and DEGs provide potential targets to further study breeding strategy to enhance NDV resistance in chickens.
ARTICLE | doi:10.20944/preprints202008.0014.v1
Subject: Biology, Animal Sciences & Zoology Keywords: Bengal slow loris; masked palm civet; common palm civet; conservation; forest canopy; density; Satchari National Park; tropical forest; Bangladesh
Online: 2 August 2020 (11:09:08 CEST)
Tropical forests harbor complex communities that are linked together by biotic relationships. Asian forests in particular have lost many apex predators due to habitat loss. We studied a small forest patch in northeastern Bangladesh, Satchari National Park, to determine density and diversity of nocturnal mammals and evaluate their relationships. Transects were walked from February 2015 to July 2016 and density was estimated using distance sampling. Nine species of mammals (5 arboreal and 4 ground-dwelling) were encountered. Densities of the common palm civets, Paradoxurus hermaphrodites, Bengal slow loris, Nycticebus bengalensis, were the highest (19.48 and 15.03 individuals/km2). Density of small Indian civets, large Indian civets and Indian mongoose were lower (2.31-5.55 individuals/km2). Unexpectedly, a wide range of nocturnal mammals co-existed in this forest patch, in spite of fragmentation and severe disturbance. We did not find any significant association between any of the species studied, although this could be an artifact of low sample size. Conservation in Bangladesh remains a challenge due to high human population density. Thus, strict conservation measures are needed to permit the long-term survival of these species.
ARTICLE | doi:10.20944/preprints202007.0679.v1
Subject: Biology, Animal Sciences & Zoology Keywords: Sub Clinical Mastitis; prevalence; Surf Field Mastitis Test; Jhenaidah
Online: 28 July 2020 (10:40:34 CEST)
A cross-sectional study was carried out to determine the prevalence of subclinical mastitis (SCM) among medium to large scale household dairy farms in southwestern district, Jhenaidah, Bangladesh during July to December 2019. A total of 78 (n=100) lactating cows from household dairy farms (N=32) having three or more dairy cows were selected randomly as sampled populations. Milk samples were screened for SCM by using Surf Field Mastitis Test (SFMT). The prevalence of SCM varied among farm level [71.9% (95% CI: 53.3-86.3)], individual animal level [67.9% (95% CI: 56.4-86.3)] and quarter level [29.5% (95% CI: 24.5-34.9)]. Descriptive statistics represented the farmers and farm demography, characteristics of the sampled population, and overall management feature. Random Effect Logistic Regression identified, Body Condition Score (BCS) [OR=3.8 and 2.9, at cows level and quarter level respectively (BCS-2 vs. BCS-≥3)], and breed [OR=5.1 and 2.9, at cows level and quarter level respectively (HF× Sahiwal vs. HF × Local)] as potential risk factors. This study shows that SCM is highly prevalent in the study area, which is a major threat to the dairy industry's production performance. Regular screening by SFMT, proper hygiene, improve the management system, and farmer’s awareness is required to control the disease.
ARTICLE | doi:10.20944/preprints202007.0594.v1
Subject: Biology, Animal Sciences & Zoology Keywords: Crustacea; Zooplankton; Plankton; Pacific Ocean; Larvae; DNA barcoding
Online: 24 July 2020 (14:25:19 CEST)
Crustacea larvae and adults make up a large fraction of the biomass and number of organisms in both holoplankton (organisms that spend their entire lives in the plankton) and meroplankton (organisms that spend their larval stages in the plankton). The life cycles of these animals can be studied by raising individuals and studying them longitudinally in the laboratory, but this method can be very laborious. Here we show that DNA sequencing of a small element in the mitochondrial DNA (DNA barcoding) makes it possible to easily link life-cycle phases without the need for laboratory rearing. It can also be used to construct taxonomic trees, although it is not yet clear to what extent this barcode-based taxonomy reflects more traditional morphological or molecular taxonomy. Collections of zooplankton were made using conventional plankton nets in Newport Bay and the Pacific Ocean near Newport Beach, California, and individual crustacean specimens were documented by videomicroscopy. Adult crustaceans were collected from solid substrates in the same areas. Specimens were preserved in ethanol and sent to the Canadian Centre for DNA Barcoding at the University of Guelph, Ontario, Canada for sequencing of the COI DNA barcode. From 1042 specimens, 609 COI sequences were obtained falling into 169 Barcode Identification Numbers (BINs), of which 85 correspond to recognized species. The results show the utility of DNA barcoding for matching life-cycle stages as well as for documenting the diversity of this group of organisms.
ARTICLE | doi:10.20944/preprints202007.0505.v1
Subject: Biology, Animal Sciences & Zoology Keywords: mitochondria dysfunction; reactive oxygen species; hypoxic; D-galactose; high fat diet; aging; hearing loss; mechanisms of hearing loss
Online: 22 July 2020 (07:58:12 CEST)
Age-related hearing loss (ARHL) is the most common sensory disorder in the elderly. It is associated with aging and hair cell death due to oxidative stress-induced mitochondrial dysfunction. Although transgenic mice and long-term cultures for induction of aging have been used to study ARHL, there are presently no ARHL animal models stimulated by intermittent environmental change for aging. In this study, an ARHL animal model was established by inducing continuous oxidative stress to promote short-term aging of cells, determined based on the expression of the hearing loss-induced phenotype and aging related factors in the short term. The incidence of hearing loss was significantly different among the groups subjected to intermittent hypoxic environment, high-fat diet (HFD), and injection with D-galactose. Continuous oxidative stress and HFD were factors that accelerated cellular aging. Increase in UCP2 affected oxidative stress and mitochondrial dysfunction. CDH23, SLC26A4, KCNQ4, Myo7a, and Myo6, which are ARHL-related factors, were modified by oxidative stress in cells of the hearing organ. We found that intermittent hypoxic, HFD, and galactose injection accelerated cellular aging in the short term. Thus, we anticipate that the development of this hearing loss animal model, which reflects intermittent environmental changes, will benefit future research on ARHL.
ARTICLE | doi:10.20944/preprints202007.0289.v1
Subject: Biology, Animal Sciences & Zoology Keywords: chickens; IGF-1 gene; TGFβ2 gene; DNA extraction; purification; sequencing and bioinformatics
Online: 14 July 2020 (05:17:40 CEST)
Molecular analysis is an easier means to identify and isolate a specific gene which has imperative function for growth, body composition , fat deposition, metabolic and skeletal traits as well as the molecular genetics selection on individual genes is a very efficient method to genetically improve economically important traits in chickens. Insulin- like growth factor 1 ( IGF-1)is a member of a heterogeneous group of peptides with important growth.Transforming growth factorβ ( TGF-β) belongs to a large family of growth and differentiation factors that play a pivotal role in a great variety of biological activities including morphogenesis, development and differentiation. DNA was extracted from 48 chickens sampled from three strains Lohman (17) , Sinai (24) and Gimmizah (7) IGF-1 gene and TGFβ2 gene were amplified using PCR protocol. Electrophoresis was carried out on the products of PCR , bands viewed on transilluminator. The size of IGF-1 gene was 675 bp while the size of TGFβ2 gene was 188bp. Sharp bands were purified and sequenced and used the dendrogram to show the relationships between other vertebrate species