Understanding the factors affecting semen quality is important for optimizing male reproductive efficiency. The present study aimed to evaluate the seminal quality associated with breed, age, season, and sperm counting chamber in boar ejaculates. A total of 22 sexually mature and healthy boars from maternal and sire commercial breeds were utilized as semen donors, with an average age (mean ± standard deviation) of 21.0 ± 7.2 months. Boars were housed individually in well-ventilated pens and fed a standard breeder mix. Semen samples were collected from each boar in dry and rainy season. The boars were grouped according to age into three categories: <12 months, 12-24 months, and >24 months. Only ejaculates with at least 75 % morphologically normal sperm and more than 8.5 x 109 total sperm per ejaculate were used. The samples were analyzed using the Computer-Assisted Semen Analysis (CASA) ISAS®v1 system. Results showed that younger boars (<12 months) exhibited higher total and progressive sperm motility and faster swimming patterns compared to older boars. The percentages of total motility and fast spermatozoa were increased by rainy season. The kinematics variables showed significant differences (P<0.05) between sperm counting chambers. Duroc and Landrace breeds presented spermatozoa with greater total motility, while terminal sire line breeds showed accelerated linear progressiveness (P<0.05). Overall, the impact of various factors on seminal and kinematic variables in boar ejaculates, including age, season, genetic breed composition, and sperm counting chamber enables to better understand the semen quality of boar ejaculates. This highlights the importance of optimizing swine reproductive management practices in sexually active boars.

Porcine metabolomics research has been extensively conducted, primarily focusing on conventional cohorts or animal models. However, the composition and functions of fecal metabolites in pigs at various ages, particularly in the elderly, remain poorly understood. In the present study, an untargeted metabolomics approach was employed to analyze the fecal metabolomes of pigs at three distinct age stages, with the aim of elucidating age-associated changes in metabolite composition and functionality under standardized rearing conditions. Through the untargeted metabolomic analysis of fecal samples from young (one year), middle-aged (four years), and elderly (eight years) pig cohorts, a variety of age-related metabolites were identified. For example, L-methionine sulfoxide was observed to increase with age, while cytidine-5-monophosphate exhibited a concomitant gradual decrease during the aging process. These metabolites were found to undergo alterations across multiple biological pathways, including energy metabolism, pyrimidine metabolism, lipid metabolism, and amino acid metabolism. Furthermore, the age-dependent metabolic changes were associated with known senescence biomarkers, offering new insights into the mechanistic understanding of aging in pigs.

Backyard production systems (BPS) are highly distributed in central Chile. Poultry BPS have been previously characterized; however, swine BPS have not yet been deeply characterized in central Chile. In addition, there is evidence that zoonotic pathogens, such as influenza A virus and Salmonella spp., are circulating in both poultry and pigs kept these production systems. A total of 358 BPS located in central Chile were evaluated between 2013 and 2015 by interviewing farm owners. Severe deficiencies in biosecurity measures were observed. The value chain of swine backyard production identified feeding, products and veterinary visits, and replacement or breeding animals as the main inputs to the backyard. The most common origin of swine replacements was from outside the BPS (63%). Meat and live animals (piglets and breeding animals) were identified as the main outputs of the system. In 16% of BPS, breeding animals were lent to other BPS, indicating the existence of animal and animal products movement in and out of backyard farms. Results from this study suggest that swine BPS in central Chile represent an animal-human interphase that need special attention to direct preventive measures aimed at avoiding the introduction and dissemination of animal pathogens and the emergence of zoonotic pathogens.

African swine fever (ASF) is a lethal and highly contagious transboundary animal disease with the potential for rapid international spread. Currently, there is no ASF vaccine commercially available. All infected animals must be isolated and culled immediately upon confirmation of the presence of the virus. Works leading to the rational development of protective ASF vaccines are urgently needed. Here, we generated a safe and efficacious live-attenuated vaccine (LAV) VNUA-ASFV-LAVL2 by serial passaging of a field isolate (VNUA-ASFV-05L1, genotype II) in porcine alveolar macrophage (PAMs, 65 passages) and an immortalized porcine alveolar macrophage cell line (3D4/21, 55 passages). VNUA-ASFV-LAVL2 can efficiently replicate in both PAMs and 3D4/21 cells. It provides 100% protection, even with the low dose of 10^2 HAD50, for the vaccinated pigs against the challenge of contemporary pandemic ASFV field isolate. Pigs vaccinated with this LAV at a dose range of 10^2 to 10^5 HAD50 remained clinically healthy during both the 28-day observation period of immunization and the 28-day observation period of challenge. VNUA-ASFV-LAVL2 was completely eliminated from blood by 28 days post inoculation (DPI), and from feces or oral fluids by 17 DPI. It remained a safe, stable, and attenuated phenotype after five passages in pigs. Consistently high levels of ASFV-specific IgG antibodies and significant cellular immunity were detected in vaccinated pigs before the ASFV challenge. These results indicate that the VNUA-ASFV-LAVL2 strain is a safe and efficacious LAV against the genotype II ASFV strain responsible for current ASF outbreaks in Asia.

Osteomyelitis is a significant cause of total carcass condemnation in pigs at the slaughterhouse. The decision for total condemnation of a pig carcass for osteomyelitis is often based on traditional perceptions of the risk of pyemia, leading to controversy among Official Veterinarians (OV) in the industry. This review aims to provide a more comprehensive understanding of the etiopatho-genesis of osteomyelitis in pigs, the microorganisms involved, and the risk factors. It also high-lights the urgent need for a more uniform method to evaluate osteomyelitis cases, which could significantly reduce economic losses in the industry. Lesions originating from tail-biting, tail docking, castration, teeth resection, and raw management are described as risk factors for oste-omyelitis. Osteomyelitis is caused by the entry of pathogens into the bloodstream of the animal through an open wound. The most described pathogens are Trueperella monocytogenes, Staphylo-coccus aureus, and Streptococcus spp. At slaughter, OVs condemn carcasses with osteomyelitis due to pyemia. Signs of acute disease are essential to identify pyemia cases. In chronic cases, total carcass condemnation can be avoided depending on the number of lesions and number of vertebrae af-fected. A clear overall image of the problem would help authorities in various countries adopt a more homogenous approach.

Background: An analysis of the pages of the WHO website on the influenza pandemic revealed changes in the use of the term "pandemic" that occurred in 2009 and 2011. Materials and methods: Since the cause of the change in the description of a pandemic on the WHO website in 2009 and 2011 is not clear, analyses of the literature related to the epidemics and pandemics, WHO documents, WHO website, and articles published in journals and the Internet have been done.Results: Until early May 2009, the description of the pandemic focused on "enormous numbers of cases and deaths." On May 6, 2009, a new description of the pandemic was published, focusing on the prevalence of the disease, but in 2011 it reverted to the initial one without any comments. From the perspective of the WHO document issued in 2009, the declaration of a swine flu pandemic in June 2009 seemed justified. However, considering the previous pandemic history, common sense, and the consequences of declaring a pandemic for a disease not accompanied by a high number of cases and high mortality, it was a premature move.Conclusion: Since the primary factor hindering the development of a pandemic is the effectiveness of treating infectious diseases, but not a definition of a pandemic, to minimize the likelihood of a new pandemic, it is necessary to improve the quality of special medical education and to study and adapt to modern conditions all effective drugs and methods used in the past.

Influenza viruses are important pathogens that affect human and animal health. The ecology and epidemiology of influenza viruses are very complex because they have a diverse host range. Influenza A viruses are responsible for serious epidemics. Influenza B viruses cause respiratory disease in humans, seals, and pigs. Influenza C viruses are infectious to humans and pigs. Influenza D virus infects cattle, although it was first isolated from a sick pig in 2011. Only influenza A viruses are of clinical importance in pigs. Although influenza A viruses infect a variety of animal species, pigs are one of the natural hosts of these viruses. Swine influenza is a very serious respiratory disease in pigs, and its main causative agents are the H1N1, H1N2, and H3N2 subtypes, which are antigenically related to human influenza viruses. Pigs are susceptible to infection by avian and human influenza viruses and are thought to play a significant role in the ecology of human influenza. This review examines scientific articles on the zoonotic spread of swine influenza A viruses to humans, their circulation in pig populations worldwide, reverse zoonosis from humans to pigs, and the role of pigs in interspecies transmission.

The performance of 5 forecasting models was investigated for predicting nursery mortality using the master table built for 3,242 groups of pigs (~ 13 million animals) and 42 variables, which concerned the pre-weaning phase of production and conditions at placement in growing sites. After training and testing each model’s performance through cross-validation, the model with the best overall prediction results was the Support Vector Machine model in terms of Root Mean Squared Error (RMSE=0.406), Mean Absolute Error (MAE=0.284), and Coefficient of Determination (R2=0.731). Subsequently, the forecasting performance of the SVM model was tested on a new dataset containing 72 new groups, simulating ongoing and near real-time forecasting analysis. Despite a decrease in R2 values on the new dataset (R2=0.554), the model demonstrated high accuracy (77.78%) for predicting groups with high (5>%) or low (5<%) nursery mortality. This study demonstrated the capability of forecasting models to predict the nursery mortality of commercial groups of pigs using pre-weaning information and stocking conditions variables collected post-placement in nursery sites.

When swine flu vaccines and circulating influenza A virus (IAV) strains are poorly matched, vaccine-induced antibodies may not protect from infection. Highly conserved T cell epitopes may, however, have a disease-mitigating effect. The degree of T cell epitope conservation among circulating strains and vaccine strains can vary, which may also explain differences in vaccine efficacy. Here, we evaluate a previously developed conserved T cell epitope-based vaccine and determine the persistence of T cell epitope conservation over time. We used a pair-wise homology score to define conservation between the vaccine’s swine leukocyte antigen (SLA) class I and II-restricted epitopes and T cell epitopes found in 1,272 swine IAV strains sequenced between 2013 and 2017. Twenty-four of the 48 total T cell epitopes included in the epitope-based vaccine were highly conserved and found in >1,000 circulating swine IAV strains over the five-year period. In contrast, commercial swine IAV vaccines developed in 2013 exhibited declining conservation with the circulating IAV strains over the same five-year period. Conserved T cell epitope vaccines may be useful adjunct for commercial swine flu vaccines and to improve protection against influenza when antibodies are not cross-reactive.

Archaea are an essential class of gut microorganisms in humans and animals. Despite the substantial progress in gut microbiome research in the last decade, most studies have focused on bacteria, and little is known about archaea in mammals. In this study, we investigated the composition, diversity, and functional potential of gut archaeal communities in pigs by re-analyzing a published metagenomic dataset including a total of 276 fecal samples from three countries: China (n=76), Denmark (n=100), and France (n=100). For alpha diversity (Shannon Index) of the archaeal communities, Chinese pigs were less diverse than Danish and French pigs (P<0.001). Consistently, Chinese pigs also possessed different archaeal community structures from the other two groups based on the Bray-Curtis distance matrix. Methanobrevibacter was the most dominant archaeal genus in Chinese pigs (44.94%) and French pigs (15.41%), while Candidatus Methanomethylophilus was the most predominant in Danish pigs (15.71%). At the species level, the relative abundance of Candidatus Methanomethylophilus alvus, Natrialbaceae archaeon XQ INN 246, and Methanobrevibacter gottschalkii were greatest in Danish, French, and Chinese pigs with a relative abundance of 14.32%, 11.67%, and 16.28%, respectively. In terms of metabolic potential, the top three pathways in the archaeal communities included the MetaCyc pathway related to the biosynthesis of L-valine, L-isoleucine, and isobutanol. Interestingly, the pathway related to hydrogen consumption (METHANOGENESIS-PWY) was only observed in archaeal reads, while the pathways participating in hydrogen production (FERMENTATION-PWY and PWY4LZ-257) were only detected in bacterial reads. Archaeal communities also possessed CAZyme gene families, with the top five being: AA3, GH43, GT2, AA6, and CE9. In terms of antibiotic resistance genes (ARGs), the class of multidrug resistance was the most abundant ARG, accounting for 87.41% of archaeal ARG hits. Our study reveals the diverse composition and metabolic functions of archaea in pigs, suggesting that archaea might play important roles in swine nutrition and metabolism.

There is no simple method to measure glomerular filtration rate (GFR) in swine, an established model to study renal disease. We developed a protocol to measure GFR in conscious swine with the plasma clearance of iohexol. We used two groups: testing and validation, of 8 animals each. Ten milliliters of iohexol (6.47 g) were injected by the marginal auricular vein and blood samples (3 ml) were collected from the orbital sinus at different points after injection. GFR was determined considering two models: two-compartments (CL2: all samples) and one-compartment (CL1: the last six samples). In the testing group, CL1 overestimated CL2 by ~30%: CL2=245±93 and CL1=308±123 ml/mn. This error was corrected by a first order polynomial quadratic equation to CL1, which was considered the simplified method: SM=-47.909+(1.176xCL1)–(0.00063968xCL1²). SM showed narrow limits of agreement with CL2, and a concordance correlation of 0.97 and a total deviation index of 14.73%. Similar results were obtained for the validation group. This protocol is reliable, reproducible, can be performed in conscious animals, uses a single dose of the marker, and requires a reduced number of samples avoiding urine collection. Finally, it portends a significant improvement in animal-welfare conditions and handling necessities in experimental trials.

المستخلص تصنيع لقاح مكون من الحمض النووي الريبوزي الرسول الخاص بالبروتينات المسئولة عن انتاج البروتينات نيورامنيديز والهيم اجليوتونين لفيروسي الانفلونزا الخنازير والطيورمحاط بجزيئات دهنية بحجم النانوميتر عدوى انفلونزا الخنازير وانفلونزا الطيور مشكلة خطيرة تؤرق العالم كله هذه الايام خصوصا انها مميتة لكثير من اطفال العالم . بعد فترة التعافي من الاصابة تتكون المناعة المكتسبة ضدها لمدة عام تقريبا. هذه الانفلونزا تسبب تسمم فيروسي في الدم بالاضافة للالتهاب الرئوي وهذامن بين اكبرالاسباب للوفاة بين الاطفال قبل سن عامين عالميا بشكل مباشر. الهدف من الدراسة هو انتاج لقاح الحمض النووي الريبوزي الرسول من نوع الام بالنانوتكنولوجي لبروتينات الهيم اجليوتونين والنيورامنيديز الموجودة على سطح هذه الفيروسات التي تساعدتها في الاتصاق بالعائل الذي ستتكاثر داخل خلاياه بالجهاز التنفسي. هذه الدراسة دراسة استكشافية ومن خلالها استطاعنا تصميم هذا اللقاح باستعمال نظام توصيل للقاح بالنانوتكنولوجي حجم جزيئاته 70نانوميمتر. اللقاح الجديد اظهر فاعلية بنسبة 60% واظهر فاعلية حيوية مقاربة واثار جانبية اقل بالمقارنة باللقاحات المتوفرة حاليا بالسوق الدوائي ضد هذا النوع من الفيروسات الوبائية. ABSTRACT Background: Worldwide, Swine and Avian influenza, or the H1N1 and H5N1 serotypes of the Influenza A virus, is a highly contagious illness. In recent years, the global incidence of mortality rate has surpassed 284,000 fatalities. Aim of the study: The production of mRNA vaccine lipid nanoparticles (LNP) to combat the deadly strains of Avian and Swine influenza (H1N1 and H5N1 of the influenza A serotype). Methodology: Polymerase chain reaction (PCR) was used to clone genes of interest encoding Haemagglutinin (HA) and Neuraminidase (NA) of Influenza A virus strains (H5N1 and H1N1) known as Avian flu and Swine flu, respectively. The genes were then inserted using the EcoR1, BAM Hind II restriction endonuclease type II, and Ligase enzyme into the pET-21(+) transcription vector plasmid. On the other hand, the hybrid transcription vectors were mixed with T7 RNA polymerase and ribonucleotides within a test tube after being linearized using EcoR II. Following their purification via reversed-phase high-performance liquid chromatography (RP-HPLC), the mRNAs of the HA and NA were encased in 70 nm LNPs and subjected to reverse-phase high-performance liquid chromatography (RP-HPLC). Lastly, immunogenicity studies were performed in stages 1/2 of randomized human and preclinical clinical trials. Results: The bivalent vaccination had around 63% efficacy in preclinical studies and approximately 60% efficacy in randomized human clinical trials. As shown by the ELISA test, strong neutralizing antibodies against HA were prevalent. Conclusion: The current study is a promising effort because of the development of the novel LNP-mRNA vaccine, which could lower the incidence of Avian and Swine flu infectious disorders.

Salmonella Typhimurium infection in pigs is characterized by an intense inflammatory response in early infection which seems to dysregulate host physiology including the gut microbiota. Both, changes in host physiology and microbiome composition, must entail modifications within the gut environment. This study analyzes the gut metabolome in early infected pigs (48 hours after infection) and non-infected controls by untargeted metabolomics (LC–QTOF MS/MS and GC–TOF MS), supported with microbiome data (16S rRNA abundance and prediction functional analyses). The metabolomic profile in Salmonella-infected pigs differed from healthy controls in 39 of these metabolites, including eight amino acids, four bile acids and carboxylic acids, three carnitines and sugar derivates, eleven fatty acids and six other compounds. Changes in amino acids abundance and the observed decrease of proteolytic bacteria such as Lactobacillus or Bacteroides could favor the niche colonization by Desulfovibrionaceae. In addition, the decrease in short chain fatty acids and tricarboxylic acids cycle was correlated to a decrease in beneficial bacteria in infected pigs. Interestingly, we observed an increase of bile acids concentration and compounds such as succinic acid or pantothenic acid, which may boost the inflammatory response. Altogether, the results reveal how S. Typhimurium infection alters the gut microbiome composition and prompts changes in the metabolism with a potential interaction between intestinal metabolism and microbiota abundance.

DNA extraction for downstream molecular diagnostic applications can be an expensive, time-consuming process. We devised a method to quickly extract genomic DNA from environmental samples based on sodium hydroxide lysis of cells with or without capture by magnetic beads, for subsequent PCR or quantitative PCR. The final DNA extraction method using NaOH is extremely low-cost and can be completed in 10 minutes at room temperature. NaOH extraction was effective for Gram-negative and Gram-positive bacteria in samples from air, soil, sewage, food, laboratory surfaces, and chicken cloacal swabs. The NaOH extraction method was comparable to commercial kits for extraction of DNA from pig fecal samples for 16S amplicon sequencing analyses. We demonstrate that an impinger and portable pump can efficiently capture bacteria in poultry facilities for rapid DNA extraction for quantification of total bacteria and for detection of specific species using qPCR. The air sampling and NaOH extraction procedures are well-suited for routine, high throughput screening, and for metagenomic analyses for specific pathogens, even in resource-limited situations.

African Swine Fever (ASF) poses a significant threat to domestic pig populations worldwide. This study investigates the histopathological alterations in confirmed ASF-positive domestic pigs, focusing on over 100 cases from 2018 to 2021. Biological material from 118 pigs, confirmed ASF-positive via PCR examination, was histopathologically examined. Tissue samples from various organs were processed, embedded, and analyzed using hematoxylin and eosin staining. Immunohistochemistry (IHC) was employed to detect the major capsid protein (p72) of the ASF virus.Histological examination revealed characteristic ASF lesions, including severe hemorrhages, lymphoid depletion, and inflammatory infiltrates. Vascular anomalies leading to hemorrhagic events were widespread. Microthromboses were prevalent in lymph nodes and kidneys. IHC detected p72 in various organs, with tonsils consistently showing the highest viral presence.This comprehensive study underscores the pivotal role of lymph nodes, spleen, and kidneys in ASF pathogenesis, highlighting prominent vascular involvement and hemorrhagic manifestations. The study establishes the efficacy of IHC in detecting ASF virus, even in autolyzed tissue samples. Tonsils emerge as a consistent epicenter of viral presence, shedding light on the intricate pathogenesis of ASFV.

The rapid expansion of livestock production sector to meet the world population demand is posing a big challenge to environmental sustainability. Plant-based feed additives extracted from food by-products could potentially result in a double outcome: improving animal zootechnical performances and mitigating environmental impacts of meat production chains. This presented study was carried out to assess the environmental impacts of the supplementation of a commercial Citrus extract feed additive (CEFA) in swine and broiler farming. Life cycle assessment (LCA) was applied to assess the impact of manufacturing and distributing 1 bag of 25 kg of CEFA and its use in feed in broiler and swine productions. With regard to CEFA manufacturing and distribution, results showed that the majority of impact came from the production of CEFA ingredients, accounting for 70% impact generated. The remaining 30% effect was divided to transportation to customer (25%), CEFA packaging (3%), CEFA manufacture, and production loss (2%). When enlarging the scope, the use of CEFA in piglet and broiler diets showed improving the measured environmental indicators, compared to such standard systems. Indeed, feed supplemented CEFA have demonstrated in elevating zootechnical performances, hence saving feed consumed. Consequently, that helped reduce environmental issues from animal feed ingredients’ agriculture. To be more specific, the supplementation of 25 kg of CEFA in feed led to a reduction of 6 tons of CO2 emitted along the life cycle of broiler production and 5 tons in the case of fattening pig. 0.7 hectares of land used had been saved as well as, 201 m3 of water (broiler) and 82 m3 (swine) were economized. Environmental performance assessment showed the interest in using this CEFA in swine and broiler diets to mitigate the environmental impacts.

Particulate matter (PM) represents an air quality management challenge for confined swine production systems. Because of the limited space and ventilation rate, PM can reach relatively high concentrations in swine barns. PM in swine barns possesses different physical, chemical, and biological characteristics than that in the atmosphere and other indoor environments. As a result, it exerts different environmental and health effects and creates some unique challenges regarding PM measurement and mitigation. Numerous research efforts have been made, generating massive data and information. However, relevant review reports are sporadic. This study aims to provide an updated comprehensive review of swine barn PM, focusing on publications since 1990. It covers various topics, including PM characteristics, sources, measurement methods, and in-barn mitigation technologies. Since PM in swine barns is of primarily biological origins, bioaerosols are reviewed in great detail. Relevant topics include bacterial/fungal counts, viruses, microbial community composition, antibiotic-resistant bacteria, antibiotic resistance genes, endotoxins, and (1→3)-β-D-glucans. For each topic, existing knowledge is summarized and discussed and knowledge gaps are identified. Overall, PM in swine barns is complicated in chemical and biological composition and highly variable in mass concentrations, size, and microbial abundance. Feed, feces, and skins constitute the major PM sources. Regarding in-barn PM mitigation, four technologies (oil/water sprinkling, ionization, alternation of feed and feeders, and recirculating air filtration) are dominant. However, none of them have been widely used in commercial barns. A collective discussion of major knowledge gaps and future research needs is offered at the end of the report.

African swine fever virus (ASFV) is a large enveloped DNA virus that causes a highly pathogenic hemorrhagic disease in both domestic pigs and wild boars. The ASFV genome contains a double-stranded DNA encoding more than 150 proteins. The ASFV possesses only one protease pS273R, which is important for virion assembly and host immune evasion. Therefore, the specific monoclonal antibody (mAb) against pS273R is useful for ASFV research. Here, we generated two specific anti-pS273R mAbs named as 2F3 and 3C2, both of which were successfully applied for ELISA, Western blotting and Immunofluorescence assays. Further, we showed that both 2F3 and 3C2 mAbs recognize a new epitope of N terminal 1-25 amino acids of pS273R protein, which is highly conserved across different ASFV strains including all genotypes I and II strains. Based on the recognized epitope, an indirect ELISA was established and was effective in detecting antibody during ASFV infection. To conclude, the specific pS273R mAbs and corresponding epitope identified will strongly promote ASFV serological diagnosis and vaccine research.

Pigs are a social species and they stablish hierarchies for a better use of resources and to reduce conflicts. However, in pig production, the opportunities for growing can differ in dominant and subordinate animals. In the present study a system was tested to perform a Dominance versus Subordinate test in growing pigs to investigate how the hierarchy affects feeding behaviour, growth and gut microbiota assessed in faeces. 64 animals housed in eight different pens were used, with four castrated males and four females in each one, weighing 18kg at arrival and maintained during the whole growing period, until 140 kg. Three stool samplings were obtained from the animals directly from the anus to avoid contamination of the faeces on 58, 100 and 133 days after the start of the study to investigate the microbiota composition. The dominant animals had higher gains during the growing period that the subordinate. In addition, they were performing more visits to the feeder throughout the day. Differential abundance patterns were observed in five bacterial genera, Oliverpabstia, Peptococcus, and Faecalbacterium being more abundant in dominant animals and Holdemanella and Acetitomaculum being overrepresented in subordinate ones. This microbial biomarker accurately classified dominant versus subordinate groups of samples with an AUC of 0.92.

Because the application of antibiotic growth promoters (AGP) causes accelerated adverse effects on the animal diet, the scientific community has taken progressive steps to enhance sustainable animal productivity without using AGP in animal nutrition. Organic acids (OAs) are non-antibiotic feed additives and a promising feeding strategy in the swine and broiler industry. Mechanistically, OAs improve productivity through multiple and diverse pathways: (a) reduction of pathogenic bacteria in the gastrointestinal tract (GIT) by reducing the gut pH; (b) boosting the digestibility of nutrients by facilitating digestive enzyme secretion and increasing feed retention time in the gut system; (c) having a positive impact and preventing meat quality deterioration without leaving any chemical residues. Recent studies have reported the effectiveness of using encapsulated OAs and synergistic mechanisms of OAs combinations in swine and broiler productivity. On the other hand, the synergistic mechanisms of OAs and the optimal combination of OAs in the animal diet are not completely understood, and further intensive scientific explorations are needed. Moreover, the ultimate production parameters are not similar owing to the type of OAs, concentration level, growth phase, health status of animals, hygienic standards, and environmental factors. Thus, those factors need to be considered before implementing OAs in feeding practices. In conclusion, the current review evaluated the basics of OAs, mode of action, novel strategies to enhance utilization, influence on growth performances, nutrient digestibility, quality traits, and meat preservation of swine and broilers and their potential concerns regarding utilization.

Understanding African swine fever virus (ASFV) transmission is essential for strategies to minimize virus spread during an outbreak. ASFV can survive for extended time periods in animal products, carcasses, and the environment. While ASFV genome was found in the environment around infected farms, data on the virus survival in soil are scarce. We investigated different soil matrices spiked with ASFV-positive blood from infected wild boar to see if ASFV can remain infectious in the soil beneath infected carcasses. As expected, ASFV genome detection was possible over the entire sampling period. Soil pH, structure, and ambient temperature played a role for the stability of infectious ASFV. Infectious ASFV was demonstrated in specimens originating from sterile sand for at least three weeks, from beach sand for up to two weeks, from yard soil for one week, and from swamp soil for three days. Virus was not recovered from two acidic forest soils. All risk mitigation experiments with citric acid or calcium hydroxide resulted in complete inactivation. In conclusion, stability of infectious ASFV is very low in acidic forest soils but rather high in sandy soils. However, given the high variability, treatment of carcass collection points with disinfectants should be considered.

Forty-eight Pasteurella multocida isolates were recovered from porcine pneumonic lungs collected in Norwestern Spain (2017- 2019). These isolates were characterized for their minimal inhibition concentrations to twelve antimicrobial agents and for the appearance of eight resistance genes: tetA, tetB, blaROB1, blaTEM, ermA, ermC, mphE and msrE. Relevant resistance percentages were shown to teracyclines, sulphamethoxazole/trimethoprim and tiamulin, thus suggesting that P. multocida isolates were mostly susceptible to amoxicillin, ceftiofur, enrofloxacin, florfenicol, marbofloxacin and macrolides. 29.2% of isolates were resistant to more than two antimicrobials. The tetracycline resistance genes (tetA and tetB) were detected in 22.9% of the isolates, but none was positive to both simultaneously; blaROB1 and blaTEM genes were found in one third of isolates but both genes were detected simultaneously in only one isolate. ermC gene was observed in 41.7% of isolates, a percentage that decreased until 22.9% for msrE; finally, ermA was harboured by 16.7% and mphE was not found in any of them. Six clusters were established based on hierarchical clustering analysis on antimicrobial susceptibility for the twelve antimicrobials. Generally, it was unable to foresee the antimicrobial susceptibility pattern for each family and the association of each particular isolate inside the clusters established from the presence or absence of the resistance genes analyzed.

The control of classical swine fever (CSF) in endemic areas has been attempted with modified live vaccines. However, in some regions, the implementation of imperfect vaccination programs has led to a reduction in the genetic diversity of the circulating strains and a change in their virulence. Porvac® subunit vaccine has been shown to provide a rapid onset of protection against the “Margarita” strain. The aim of this study was to evaluate whether the immune response induced by Porvac® is also effective against autochthonous isolates of low, medium or high virulence. All pigs vaccinated with Porvac® were protected against the disease after the challenge. PR-11/10–3 isolate caused a very mild disease in controls, whilst Holguin_2009 isolate produced mild to moderate signs of CSF and one of the pigs died. Finally, controls inoculated with PR-2016 isolate developed moderate to severe signs of CSF and two of them died. Viral replication was detected in controls, but not in pigs immunized with Porvac®. Finally, anti-Erns antibodies were induced in five out of six control pigs but in any of the vaccinated pigs. These results support the use of Porvac® for the control and elimination of CSF in Cuba and other endemic regions.

Three-choice preferences for fermented sorghum teas (FST) were examined among 11 adult Ossabaw pigs. The objectives for experiments 1 and 2 (respectively) were to determine preference for: 1) one of three types of FST (white vs. sumac vs. roasted sumac), and 2) isocaloric control (+control; made isocaloric with sucrose), blended FST (3Tea; equal parts of white, sumac, and roasted-sumac), or a negative control (-control; distilled water; avoidance). Pigs were first conditioned to navigate a three-choice preference maze. For experiment 1, no clear preference behaviors for tea-type were observed. Pigs consumed nearly all the teas, but pigs consumed the least amount of tea in the first session. Pigs spent the most time with their heads in the bowl when 3Tea was offered, followed by +control, and the least time was -control.  Regardless of tea-type, adult pigs show a strong preference for FST, even over +control, which was sweetened water. Adult pigs likely prefer the complexity of flavors, rather than the sweetness alone.  Future projects that explore the health benefits of this sorghum formula will not be impeded by avoidance among adult pigs.

No system nor standardized analytical procedures at commercial laboratories exist to facilitate and accurately measure potential viable virus contamination in feed ingredients and complete feeds globally. As a result, there is high uncertainty of the extent of swine virus contamination in global feed supply chains. Many knowledge gaps need to be addressed to improve our ability to prevent virus contamination and transmission in swine feed. This review summarizes the current state of knowledge involving: 1) the need for biosecurity protocols to identify production, processing, storage, and transportation conditions that may cause virus contamination of feed ingredients and complete feed, 2) challenges of measuring virus inactivation, 3) virus survival in feed ingredients during transportation and storage, 4) minimum infectious doses, 5) differences between using a Food Safety Objective versus a Performance Objective as potential approaches for risk assessment in swine feed, 6) swine virus inactivation from thermal and irradiation processes, and chemical mitigants in feed ingredients and complete feed, 7) efficacy of virus decontamination strategies in feed mills, 9) benefits of functional ingredients, nutrients, and commercial feed additives in pig diets during a viral health challenge, and 10) considerations for improved risk assessment models of virus contamination in feed supply chains.

Strengthening the analysis and risk assessment of the pig epidemic will help to better prevent and mitigate epidemic risks and promote high-quality development of the pig industry. Based on the systematic grasp of the situation of live pig epidemics, a risk assessment index system was constructed, and the spatial and temporal variation characteristics of the risk of pig epidemics in China were explored by the entropy method. In recent years, the overall trend in pig epidemics in time first increased and then decreased, and in space, the trend in the acceleration of the spread across the country continued to weaken. China still faces challenges, such as many types and a wide range of diseases, large total livestock breeding and weak epidemic prevention and control capacity, and a large risk of introduction of foreign animal epidemics. The spatial and temporal variation in the pig epidemic risk was obvious; there was one high-risk area, two higher-risk areas and 10 medium-risk areas in recent years, and the epidemic risk was highest in Beijing, Hainan, Liaoning, Tibet and Zhejiang. However, there were significant differences in the regional distribution of the risk of pig epidemics in different years. To further build a secure “defense system” for the high-quality development of the pig industry, it is recommended to improve the monitoring and early warning system of pig epidemic risk, perfect the pig epidemic prevention and control system, and strengthen the regional collaboration mechanism of epidemic prevention and control.

Piggery wastewater (PWW) is characterized by its high concentrations of organic matter and ammonium, and by their odour nuisance. Traditionally, PWW has been treated in open anaerobic lagoons, anaerobic digesters and activated sludge systems, which exhibit high greenhouse gas emissions, a limited nutrients removal and a high energy consumption, respectively. Photosyn-thetic microorganisms can support a sustainable PWW treatment in engineered photobioreactors at low operating costs and with an efficient recovery of carbon, nitrogen and phosphorous. These microorganisms are capable of absorbing solar irradiation through the photosynthesis process to obtain energy, which is used for their growth and associated carbon and nutrients assimilation. Purple phototrophic bacteria (PPB) represent the photosynthetic microorganisms with the most versatile metabolism in nature, while microalgae are the most studied photosynthetic microor-ganisms in recent years. This review describes the fundamentals, symmetry and asymmetry of PWW treatment using photosynthetic microorganisms such as PPB and microalgae. The main photobioreactor configurations along with the potential of PPB and microalgae biomass valori-zation strategies are also discussed.

African swine fever (ASF) is one of the most important and devastating viral diseases in wild boar and domestic pigs worldwide. In the absence of vaccines or treatment options, early clinical detection is key and requires sound knowledge of disease characteristics. To provide practitioners and state veterinarians with detailed information, the objective of the present study was to characterize the ASF virus (ASFV) isolate “Belgium 2018/1” in subadult and weaning domestic pigs. To this end, two animal trials were performed. Trial A included eight subadult domestic pigs and trial B five weaner pigs. In general, clinical signs and pathological lesions were in line with previous studies utilizing highly virulent ASF genotype II viruses. However, in trial A, four subadult domestic pigs survived and recovered pointing to an age dependent outcome. The long-term fate of those survivors remains under discussion and would need further investigations.

Background An outbreak of African swine fever (ASF) in China in 2020 has led to an unprecedented shortage of fraxiparin. Most patients, especially those kept in hospital for surgery, are currently treated with prophylactic anticoagulation (AC). In search of alternatives for fraxiparin, we found no sufficient data on alternatives for neurosurgical patients, such as tinzaparin of European origin. We compared fraxiparin and tinzaparin concerning adverse events (bleeding versus thromboembolic events) in neurosurgical patients. Methods Between 2012 and 2018, 517 neurosurgical patients with benign and malignant brain tumors as well as 297 patients with subarachnoid hemorrhage (SAH) were treated in the Department of Neurosurgery, University Hospital Leipzig receiving prophylactic anticoagulation within 48 hours. In 2015, prophylactic anticoagulation was switched from fraxiparin to tinzaparin throughout the university hospital. In a retrospective manner, the frequency and occurrence of adverse events (rebleeding and thromboembolic events) in connection with the substance used was analyzed. Statistical analysis was performed using Fisher’s exact test and the chi-squared test. Results Rebleeding rates were similar in both fraxiparin and tinzaparin cohorts in patients being treated for meningioma, glioma, and SAH combined (8.8 vs 10.3%). Accordingly, the rates of overall thromboembolic events were not significantly different (5.5% vs 4.3%). The severity of rebleeding did not vary. There was no significant difference among subgroups when compared for deep vein thrombosis (DVT) or pulmonary embolism (PE). Conclusion In this retrospective study, tinzaparin seems to be a safe alternative to fraxiparin for AC in patients undergoing brain tumor surgery or suffering from SAH.

African swine fever (ASF) is a viral infection of pigs and represents a major threat to animal health and trade. Due to the high tenacity of the causative virus also in carcasses of wild boar, contacts of wild boar with infectious carcasses are regarded an important driver of the so-called habitat cycle. The latter is believed to play a major role in maintaining the present ASF situation in wild boar in Europe. Therefore, search campaigns and timely removal and disposal of carcasses are considered important disease control approaches. If timely disposal is not feasible due to logistic reasons, deterrence of wild boar could be a provisionary option. The performance of seven deterrents (physical and chemical) was tested in a forest near Greifswald, Germany. Carcasses as entities of attraction for wild boar were substituted by luring sites. It could be demonstrated that certain physical (LED-Blinkers, aluminum stripes) and chemical (Wildschwein-Stopp™, Hukinol™) deterrents are capable of reducing significantly the odds of wild boar contacts to one third. It is recommended to carry a choice of the aforementioned, reasonable and easy to apply deterrents, when carcass search campaigns are launched in case of an outbreak of ASF in wild boar.

African swine fever (ASF) continues to spread in Africa, Europe, Asia, and the island of Hispaniola increasing the need to develop more streamlined and highly efficient surveillance and diagnostic capabilities. One way to achieve this is by further optimization of already established standard operating procedures to remove bottlenecks for high-throughput screening. Real-time polymerase chain reaction (RT-PCR) is the most sensitive and specific assay available for the early detection of ASF, but it requires high quality nucleic acid extracted from the samples. Whole blood from live pigs and spleen tissue from dead pigs are the preferred samples for ASF RT-PCR. Whole blood can be used as is in nucleic acid extraction, but spleen tissues require an additional homogenization step. In this study, we compared the homogenates and swabs prepared from 52 spleen samples collected from pigs experimentally inoculated with highly and moderately virulent ASF virus strains. The results show that not only are the spleen swabs more sensitive when executed with a low-cell-count nucleic acid extraction procedure followed by RT-PCR assays, they also increase the ability to isolate ASFV from positive spleen samples. Swabbing is a convenient, simpler, and less time-consuming alternative to tissue homogenization. Hence, we recommend spleen swabs over tissue homogenates for high-throughput detection of ASFV by real-time PCR.

Background: Uncontrolled hemorrhage is a major preventable cause of death in patients with trauma. However, the majority of large animal models of hemorrhage have utilized controlled hemorrhage rather than uncontrolled hemorrhage to investigate the impact of immunopathy and coagulopathy on multi-organ failure (MOF) and mortality. This study evaluates these alterations in a severe porcine-controlled and uncontrolled hemorrhagic shock (HS) model. Methods: Anesthetized female swine underwent controlled hemorrhage (22 ml/kg) and uncontrolled hemorrhage by partial splenic resection followed with or without lactated Ringer solution (LR) or Voluven® resuscitation. Blood chemistry, physiologic variables, systemic and tissue levels of complement proteins and cytokines, coagulation parameters, organ function, and damage were recorded and assessed. Results: HS resulted in systemic and local complement activation, cytokine release, metabolic acidosis, hyperkalemia, MOF, and no animal survival. Resuscitation with LR and Voluven® after HS improved hemodynamic parameters (MAP and SI), metabolic acidosis, hyperkalemia, and survival but resulted in increased complement activation and worse coagulopathy. Compared with the LR group, the HS animal treated with Voluven® had worse dilutional anemia, coagulopathy, renal and hepatic dysfunction, increased myocardial complement activation and renal damage, and decreased survival rate. Conclusions: Hemorrhagic shock triggers early immunopathy and coagulopathy that appear associated with MOF and death. This study indicates that immunopathy and coagulopathy are therapeutic targets that may be addressed with a high-impact adjunctive treatment to conventional resuscitation.

African swine fever has damaged the foundation of China’s hog industry, caused a serious decline in hog production, highlighted the contradiction between supply and demand in the pork market, and led to major economic and social impacts. This study measured the industrial resilience of 31 Chinese provinces which faced the African swine fever shock and its spatial and temporal differentiation characteristics from 2018-2021 in terms of the two dimensions of resistance and recoverability. Using Geodetector, the study explored the key factors influencing the resilience of China’s hog industry. The results showed that 2018-2019 and 2020-2021 were the resistance and recovery periods of the hog industry under the African swine fever shock, respectively, with poor resilience characterizing the resistance period and improved resilience exemplifying the recovery period. At the early stage of the African swine fever outbreak, Tianjin, Shanxi, Guangxi and Yunnan had robust resistance in the hog industry, and the slaughter rate, economic level, mortality rate, carcass weight and culling rate showed the highest explanation for the resistance. At the most severe stage of the outbreak, the resistance of the hog industry in all provinces was generally poor, and the slaughter rate, per capita consumption and scale level showed the highest explanation for the resistance. In the period of rapid recovery of hog production, the recoverability of each province was very strong, and the industrial structure, culling rate, economic level and resource carrying capacity showed the highest explanation for the recoverability. In the reasonable adjustment period of hog production capacity, the recoverability based on the breeding sow inventory in 13 provinces, including Henan, Shandong and other large hog breeding provinces, was negative, and the scale level, slaughter rate, per capita consumption and resource carrying showed the highest explanation for the recoverability. It is recommended to take measures to enhance the resilience of the hog industry, strengthen the prevention and control of hog epidemics, improve the monitoring and early warning mechanism, and strive to enhance the ability of the hog industry to cope with major animal epidemics.

The objective of this study was to evaluate the potential benefits of feeding spray-dried porcine plasma (SDPP) to pigs infected with African swine fever virus (ASFV). Two groups of twelve weaned pigs each were fed with CONVENTIONAL or 8% SDPP enriched diets. Two pigs (trojans)/group) were injected intramuscularly with the pandemic ASFV (Georgia 2007/01) and comingled with the rest of the pigs (1:5 trojan:naïve ratio) to simulate a natural route of transmission. Trojans developed ASF and died within the first week after inoculation but contact pigs did not develop ASF, viremia or seroconversion. Therefore, three more trojans per group were introduced to optimize the ASFV transmission (1:2 trojan:naïve ratio). Blood, nasal and rectal swabs were weekly harvested and at end of the study, ASFV-target organs collected. After the second exposure, rectal temperature of conventionally fed contact pigs increased >40.5˚C while fever was delayed in the SDPP contact pigs. Additionally, PCR Ct values in blood, secretions and tissue samples were significantly lower (P<0.05) for CONVENTIONAL compared to SDPP contact pigs. Under these study conditions, contact exposed pigs fed SDPP had delayed ASFV transmission and reduced virus load, likely by enhanced specific T-cell priming after the first ASFV-exposure.

The impact of cyclic heat stress (CHS) and lighting events on pig feeding behavior (FB) was investigated. The FB of 90 gilts was recorded in real time under two ambient temperatures (AT): thermoneutrality (TN, 22 °C) or CHS (22°/35 °C). The day was divided into four periods: PI (06-08 h); PII (08-18 h); PIII (18-20 h); and PIV (20-06 h). Automatic and Intelligent Precision Feeders recorded each feed event for each pig. An estimated meal criterion (49 minutes) was used to calculate the FB variables. Feed behavior in both ATs followed a circadian pattern. The CHS reduced the feed intake by 6.9%. Pigs prioritized feed intake during the coolest hours of the day; however, nocturnal cooling did not allow the pigs to compensate for the reduced meal size due to CHS. The highest meal size and most of the meals were observed during the lighting-on period. Pigs reduced their interval between meals during periods II and III. The lighting program increased the meal size when the lights were switched on and reduced the meal size when the lights were switched off. Thus, the dynamics of the FB were largely influenced by AT, while the meal size was affected by the lighting program.

Porcine pleuropneumonia (PPP) is one of the main causes leading to massive losses in the pig industry, with high economic impacts. Among different etiological agents, Actinobacillus pleuropneumoniae (APP) is responsible for severe fibrinous-necrotizing pleuropneumonia. A total of 19 different APP serotypes are currently recognized. This study aimed to identify APP serotypes isolated from pneumonic lesions in naturally infected and dead pigs in the Piedmont Region and to describe lesions. A total of 107 dead pigs with a suspected PPP diagnosis were included in this study. Lungs were evaluated using gross pathology scoring systems, APP isolation and serotypes identification by multiplex PCR were conducted. Gross lung lesions were mainly represented by fibrinous pneumonia and pleuropneumonia. APP was isolated in 21/107 (19.6%) samples. On the contrary, PCR indicated APP DNA presence in 53/107 (49.5%) of lung samples. The most observed serotypes were serotype 2 in 24/53 (45.3%) and serotype 6 in 13/53 (24.5%) samples. Moreover, multiplex PCR results suggested a coinfection of different serotypes in 5 samples. This study highlights the relevance of the different techniques application (i.e. bacteriological culture, PCR, gross and histopathology) for improving the identification of APP infections.

The B169L protein (pB169L) of African swine fever virus (ASFV) is a structural protein with an unidentified function during the virus replication. The sequences of the B169L gene and the downstream B438L gene are separated by short intergenic regions. However, the regulatory mode of the gene transcription remains unknown. Here, we identified two distinct promoter regions and two transcription start sites (TSS) located upstream of the open reading frame (ORF) of B438L. Using the promoter reporter system, we demonstrated that the cis activity of the ORF proximal promoter exhibited significantly higher levels compared with that of the distal promoter located in the B169L gene. Furthermore, transfection with the plasmids with two different promoters for B438L could initiate the transcription and expression of the B438L gene in HEK293T cells, and the cis activity of the ORF proximal promoter also displayed the higher activities compared with the distal promoter. Interestingly, the B438L distal promoter also initiated the transcription of the alternatively spliced B169L mRNA (B169L mRNA2) encoding a truncated pB169L (tpB169L) (amino acids 92–169), and the gene transcription efficiency was increased upon mutation of the initiation codon located upstream of the alternatively spliced B169L gene. Taken together, we demonstrated that the distal promoter of B438L gene initiates the transcription of both the B438L mRNA and B169L mRNA2. Comprehensive analysis of the transcriptional regulatory mode of the B438L gene is beneficial for the understanding of the association of B438L protein and pB169L and the construction of the gene-deleted ASFV.

African swine fever (ASF) has evolved from an exotic animal disease to a threat to global pig production. An important avenue for wide-spread transmission of animal diseases is the dissemination of viruses through boar semen used for artificial insemination (AI). In this context, we investigated the role of male reproductive organs in ASF. Mature domestic boars and adolescent wild boar inoculated with different ASF virus strains were investigated by means of virological and pathological methods. Additionally, electron microscopy was employed to investigate in vitro inoculated sperm. Viral genome, antigen and infectious virus could be found in all gonadal tissues and accessory sex glands. The viral antigen and viral mRNAs were mainly found in mononuclear cells of the respective tissues. However, some other cell types, including Leydig, endothelial and stromal cells were also found positive. Using RNAScope, p72 mRNA could be found in scattered halo cells of the epididymal duct epithelium which could point to disruption of the barrier. No direct infection of spermatozoa was observed by immunohistochemistry or electron microscopy. Taken together, our results strengthen the assumption that ASFV can be transmitted via boar semen. Future studies are needed to explore excretion dynamics and transmission efficiency.

Swine influenza A viruses (IAVsw) are important causes of disease in pigs but also constitute a public health risk. IAVsw strains show remarkable differences in pathogenicity. We aimed to generate airway organoids from the porcine lower respiratory tract and use these to establish well-differentiated airway epithelial cell (WD-AEC) cultures grown at air-liquid interface (ALI) for in vitro screening of IAVsw strain virulence. Epithelial cells were isolated from bronchus tissue of juvenile pigs, and airway organoids were cultured in extracellular matrix in medium with human growth factors. Single cell suspensions of these 3D organoids were seeded on transwell filters and differentiated at ALI to form a pseudostratified epithelium containing ciliated cells, mucus-producing cells and tight junctions. Inoculation with a low dose of IAVsw in a low volume inoculum resulted in virus replication without requiring addition of trypsin, and was quantified by detection of viral genome loads in apical washes. Interestingly, inoculation of an H3N2 strain known to cause severe disease in pigs induced greater reduction in trans-epithelial resistance and more damage to tight junctions than H1N2 or H1N1 strains associated with mild disease in pigs. We conclude that the porcine WD-AEC model is useful to assess the virulence of IAVsw strains.

Pig farms management is involved in multiple factors such as environmental conditions, animal behavior, weight gaining and others. In all those scenarios, it is fundamental to detect each individual and its development as it grows to reach the shipment phase. Therefore, the objective of this study was to apply an automatic individual pig detection algorithm using Deep Learning Techniques, in frames taken from videos recorded during the weaning phase in real production conditions of small and medium producers in tropical conditions in Colombia. The weaning phase was selected because individual detection in each cage is more difficult due to environmental adaptations that piglets undergo after nursing. Piglets tend to cluster together during this phase, making image processing more challenging compared to the fattening phase, where animals are more isolated. The You Only Look Once (YOLOv7) algorithm and VGG Image Annotator were used to detect individuals in two pens separated by gender. All animal postures, lighting conditions were considered. The Precision, Recall and mAP metrics were used to assess the model performance with different confidence values. A precision of 98.3% in a threshold of 0.9 was reached, Recall of 98.5% in a threshold of 0.85. Data augmentation techniques were important to increase the dataset and to have a more realistic model for individuals’ detection. The method used allowed us to identify all the complete pigs’ bodies under different posture and lighting situations.

The objective of the present report is the dissemination of information acquired during the application of ISO 6579-1:2017 for the isolation of Salmonella spp. from swine samples. ISO 6579-1:2017 is the officially recommended by the EU protocol for the isolation of Salmonella spp., aiming in the harmonization of effective successful control of Salmonella infection in food producing animals. Successful control of animal salmonellosis is highly dependent on the sensitivity of the biochemical methods reliably detecting the presence of the pathogen in various stages of food production. Application of ISO 6579-1:2017 resulted in the isolation of twelve Salmonella spp. and eighteen other members of the family of Enterobacteriaceae biochemically and antigenically resembling salmonellae. The findings necessitated in the depth evaluation of the culture media xylose-lysine-desoxycholate agar (XLD), Salmonella-Shigella Agar (SS), Brilliant Green Agar (BGA), Salmonella Chromogenic Agar (SCA), Triple Sugar Iron (TSI) and modified semi-solid Rappaport–Vassiliadis (MSRV) agar. The evaluation showed that these culture selective media differed significantly in their performance regarding the isolation of Salmonella from swine samples. It was concluded that the presence of atypical Salmonella strains negatively affects the prevalence of Salmonella, thus the identification of carrier pigs, eventually affecting the efficiency of control programs. Thus, doubtful results require additional biochemical testing for confirming the accuracy of such universally recommended isolation methods.

Contrary to pig farming in developed western countries, in a large part of the world, pigs are still traditionally kept, in small backyard farms, usually for family needs. Their main characteristics are low biosecurity, swill feeding, natural breeding and uncontrolled trade. Given the high number of backyard farms in Serbia and the risk they are thought to pose to intensive pig farming, the main aim of this study was to evaluate the prevalence of major viral diseases of swine among traditionally kept pigs in small holdings with low biosecurity. For this investigation, 222 serum samples from 69 backyard holdings were randomly selected and tested for PRRS, Aujeszky`s disease, PPV infection and Swine influenza by ELISA. The herd-level seroprevalence of PRRS, Aujeszky`s disease and PPV infection was 2.9%, 27.5% and 37.7%, respectively. Swine influenza seroconversion was not confirmed in any of the tested holdings. Despite widely distributed PPV and AD in backyard farms in Serbia, almost 50% of them are still negative for the all-tested disease. The backyard farms must be monitored, and owners educated as their role in eradication programs and obtaining country-free status may be crucial.

The dimorphic, soil-dwelling fungus Coccidioides is the causative agent of coccidioidomycosis and is a reemerging pathogen of human health concern. Despite increasing incidence and isolations of the organism outside of its theoretical environmental range, much of Coccidioides epidemiology, including complete host range, remains unknown. Feral swine (Sus scrofa) are an invasive species distributed throughout the United States that may play a role in fungal ecology due to their behavioral associations with soil. To address this, we evaluated seven feral swine for susceptibility to Coccidioides. After intranasal inoculation with 106.2 arthroconidia of C. posadasii, no pigs displayed signs of clinical disease, nor did any exhibit antibodies by AGID testing. Histopathologic evaluation did not reveal the presence granulomatous lesions, and fungal spherules or endospores were not observed in any of the tissue sections examined, although individuals had significant comorbidities, most notably Metastrongylus nematodes in the lungs. Despite the absence of lesions and organisms histologically, C. posadasii was isolated by culture from the lung and mediastinal lymph node of two pigs, indicating active infection. These results suggest that feral swine are mildly susceptible to acute Coccidioides infection and may aid in fungal dispersal due to the presence of spherules in tissues post-mortem.

This study aimed to evaluate the effects of feeding spray-dried porcine plasma (SDPP) on the protection afforded by the BA71∆CD2 African swine fever virus (ASFV) vaccine prototype. Two groups of pigs ac-climated to diets without or with 8% SDPP were intranasally inoculated with 105 plaque forming units (PFU) of live attenuated ASFV strain BA71∆CD2 and three weeks later left in direct contact with pigs infected with the pandemic Georgia2007/01 ASFV strain. During the post-exposure (pe) period, 2/6 from the conventional diet group showed a transient peak rectal temperature >40.5ºC before day 20 pe and some tissue samples collected at 20 d pe from 5/6 were PCR+ for ASFV, albeit showing Ct values much higher than Trojan pigs. Interestingly, the SDPP group did not show fever, neither PCR+ in blood nor rectal swab at any time pe and none of the postmortem collected tissue samples were PCR+ for ASFV. Differential serum cytokine profiles among groups at vaccination, and a higher number of ASFV-specific IFNϒ-secreting T-cells in pigs fed with SDPP soon after the Georgia2007/01 encounter, confirmed the relevance of Th1-like responses in ASF pro-tection. We believe that our result show that nutritional interventions might contribute to improve future ASF vaccination strategies.

African swine fever (ASF) is a pandemic threat to the global pig industry and wild suids. A safe and efficacious vaccine could monumentally assist in disease eradication. In the past years, promising live attenuated vaccine candidates emerged in proof-of-concept experiments, among them, “ASFV-G-∆MGF”. In our study, we tested the vaccine candidate in three animal experiments intramuscularly in domestic pigs one orally in wild boar. Further, a macrophage-grown vaccine virus and a virus grown on permanent cells could be employed. Irrespective of the production system of vaccine virus, a two-dose intramuscular immunization could induce close to sterile immunity with full clinical protection against challenge infection. After oral immunization, 50% of the vaccinees seroconverted and all responders were completely protected against subsequent challenge. All non-responders developed ASF upon challenge with two acute lethal infections and two mild and transient courses. The latter results show a lower efficiency after oral administration that would have to be taken into consideration when designing vaccination-based control measures. Our findings suggest that “ASFV-G-∆MGF” could help to contain the disease under an appropriate vaccination campaign. Further research is needed to characterize safety aspects and define possible improvements of oral efficiency.

African swine fever (ASF) is one of the major threats to pig production, and real-time PCR (qPCR) protocols are integral part of ASF laboratory diagnosis. With the pandemic spread of ASF, commercial kits have risen on the market. In Germany, the kits have to go through an approval process and thus, general validation can be assumed. However, they were never compared to each other. In this study, 12 commercial PCR kits were compared to an OIE recommended method. Samples representing different matrices, genome loads, and genotypes were included in a panel that was tested under diagnostic conditions. The comparison included user-friendliness, internal controls, and the time required. All qPCRs were able to detect ASFV genome in different matrices across all genotypes and disease courses. With one exception, there were no significant differences when comparing the overall mean. The overall specificity was 100 % [95 % CI 87.66 - 100], and the sensitivity was between 95 % and 100 % [95 % CI 91.11 - 100]. As can be expected, variability concerned samples with low genome load. Concluding, all tests were fit for purpose. The test system can therefore be chosen based on compatibility and prioritization of the internal control system.

Endemically infected European wild boar are considered a major reservoir of African swine fever virus in Europe. While high lethality was observed in the majority of field cases, strains of moderate virulence occurred in the Baltic States. One of these, “Estonia 2014”, led to a higher number of clinically healthy, antibody-positive animals in the hunting bag of North-Eastern Estonia. Experimental characterization showed high virulence in wild boar but moderate virulence in domestic pigs. Putative pathogenic differences between wild boar and domestic pigs are unresolved and comparative pathological studies are limited. We here report on a kinetic experiment in both subspecies. Three animals each were euthanized at 4, 7 and 10 days post infection (dpi). Clinical data confirmed higher virulence in wild boar although macroscopy and viral genome load in blood and tissues were comparable in both subspecies. The percentage of viral antigen positive myeloid cells tested by flow cytometry did not differ significantly in most tissues. Only immunohistochemistry revealed consistently higher viral antigen loads in wild boar tissues in particular 7 dpi, whereas domestic pigs already eliminated the virus. The moderate virulence in domestic pigs could be explained by a more effective viral clearance.

Chlamydia trachomatis (Ct) infections are the most common sexually-transmitted infection (STI). Despite effective antibiotics for Ct, undetected infections or delayed treatment can lead to infertility, ectopic pregnancies, and chronic pelvic pain. Besides humans, chlamydia poses similar health challenges in animals such as C. suis (Cs) in pigs. Based on the similarities between humans and pigs as well as their chlamydia species, we use pigs as a large biomedical animal model for chlamydia research. In this study, we use the pig model to develop a vaccine candidate against Ct. The vaccine candidate consists of TriAdj-adjuvanted chlamydial protease-like activity factor (CPAF) protein. We tested two weekly administration options – twice intranasal (IN) followed by twice intramuscular (IM), or twice IM followed by twice IN. We assessed the humoral immune response not only in serum by CPAF-specific IgG including antibody avidity determination but also in cervical and rectal swabs by CPAF-specific IgG and IgA ELISAs. The systemic T-cell response was analyzed upon in vitro CPAF-restimulation via IFN-γ and IL-17 ELISpots as well as intracellular cytokine staining flow cytometry. Our data demonstrate that while the IN/IM vaccination mainly led to non-significant systemic immune responses, the vaccine candidate is highly immunogenic if administered IM/IN. This vaccination strategy induced high serum anti-CPAF IgG levels with strong avidity as well as high IgA and IgG levels in vaginal and rectal swabs and in uterine horn flushes. In addition, this vaccination strategy prompted a pronounced cellular immune response: Besides inducing IL-17 production, the vaccine candidate induced a strong IFN-γ response by CD4 T cells. In the IM/IN vaccinated pigs, these cells also significantly downregulated their CCR7 expression – a sign for differentiation into peripheral tissue homing effector/ memory cells. Conclusively, this study demonstrates strong immunogenicity of the IM/IN administered TriAdj-adjuvanted Ct CPAF vaccine candidate. Future studies will test vaccine efficacy of this promising Ct vaccine candidate. In addition, this project demonstrates suitability of the Cs pre-exposed outbred pig model for Ct vaccine development. Thereby, we aim to open the bottleneck of large animal models to facilitate the progression of Ct vaccine candidates into clinical trials.

The aim of this study was to investigate the performance of the developed real-time controlled bio-liquor circulation system (BCS) on a farm-scale to optimize bioreactor operation for manure treatment, and to subsequently reduce both manure-related malodor and greenhouse gas (GHG) emissions from slurry pits. The BCS was operated sequentially as per swine manure inflow (anoxic, aerobic, and settling) circulation to the slurry pit. The duration of each operational phase was self-adjusted in real-time using a novel algorithm for detecting the control point on the oxidation-reduction potential (ORP) and pH (mV)-time profiles, the nitrogen break point (NBP), and the nitrate knee point (NKP) in the aerobic and anoxic phases, respectively. The NH₄-N in the slurry manure was thoroughly removed (100%) in the bioreactor, optimizing the duration of each operational phase by accurately detecting real-time control points. The newly-developed real-time BCS decreased the nitrogen and organic matter content in the slurry pit manure by > 70%, and the potential ammonia and methane emissions by 75% and 95%, respectively. This study highlights that the this improved BCS that utilizes ORP tracking and pH (mV)-time profiles can effectively optimize BCS operation, and thereby reduce malodor and GHG emissions from swine farms.

Candidate vaccines against African swine fever virus (ASFV) based on naturally attenuated or genetically modified viruses have the potential to generate protective immune responses, although there is no consensus on what defines a protective immune response against ASFV. Studies, espe-cially in sensible host species, focused on unravelling protective mechanisms will contribute to the development of safer and more effective vaccines. The present study provides a detailed analysis of phenotypic and functional data on cellular responses induced by intradermal immunization and subsequent boosting of domestic pigs with the naturally attenuated field strain Lv17/WB/Rie1, as well as the mechanisms underlying protection against intramuscular challenge with the virulent genotype II Armenia/07 strain. The transient mild to moderate increase of IL-8 and IL-10 in serum observed after immunization might be directly correlated with survival. Protection was also associated with a robust ASFV-specific polyfunctional memory T cell response, where CD4CD8 and CD8 T cells were identified as the main cellular sources of virus-specific IFNγ and TNFα. In parallel to cytokine response, these T-cell subsets also showed specific cytotoxic activity as evidenced by the increased expression of the CD107a degranulation marker. Along with virus-specific multifunctional CD4CD8 and CD8 T responses, the increased levels of antigen experienced cytotoxic CD4 T cells observed after challenge in immunized pigs might also contribute to control virulent infection by killing mechanisms targeting infected antigen-presenting cells. Future studies should elucidate whether the memory T-cell responses evidenced in the present study persist and provide long-term protection against further ASFV infections.

African Swine Fever (ASF) is one of the most important and serious contagious haemorrhagic viral disease of domestic pigs and wild boar, associated to high mortality rates and great sanitary and socioeconomic impact on international trade in animal and swine products. The early detection of the disease is often hampered by an inadequate surveillance. Among the surveillance strategies, passive surveillance of wild boar was considered the most effective for controlling African swine fever virus (ASFV). Otherwise, the design of a sufficiently sensitive ASF surveillance system re-quires a solid understanding of the epidemiology related to the local eco-social context, especially in absence of virus detection. Even if the number of carcasses needed to demonstrate the ASF eradication has been established, the scientific context is poor of details on protocols applied to active search the wild boar carcasses. The aim of the study was to describe the strategy applied for passive surveillance implemented in Sardinia, providing detailed information on the protocol applied in the field in term of number of people, dogs, time and space used to active search car-casses in the Mediterranean area. Using a specific tool developed to record, trace, and share on field data (GAIA Observer App), a total of 33 activities for active search the wild boar carcasses were organized during 2021-2023. Most of these activities were planned to find carcasses previously reported by hunters. A total of 24 carcasses were found, only 2 carcasses were not previously reported. The final protocol applied involved 4 people, with an average speed of 1.5 km/hour. When the carcass was reported, about 2 km of distance should be covered in about 1.5 hours to find the carcasses, and even less time was spent when the dog (not trained) was present. In conclusion, it can be stated that for searching for carcasses it is first mandatory a good collaboration with hunters or other forest visitors to report the carcasses, and involve small groups of expert people to active search the carcasses, possible with the use of hunting dogs with no special training.

To determine promethazine (PMZ) and its metabolites promethazine sulfoxide (PMZSO) and monodesmethyl-promethazine (Nor1PMZ) in swine muscle, fat, kidney, and liver, a sample preparation and high-performance liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis method were established and validated. The sample was extracted with 0.1% formic acid-acetonitrile, and the extract was purified with acetonitrile-saturated n-hexane. After concentration by rotary evaporation, the extract was re-dissolved in 0.1% formic acid-water and acetonitrile (80:20, v/v). The HPLC-MS/MS column used was a Waters Symmetry C18 (100 mm × 2.1 mm i.d., 3.5 μm), with 0.1% formic acid-water and acetonitrile as the mobile phase, and the target compounds were determined by positive ion scan and multiple reaction monitoring. PMZ and Nor1PMZ were quantified with deuterated promethazine (PMZ-d6) as the internal standard, while PMZSO was quantified by external standard method. The limits of detection (LOD) and limits of quantification (LOQ) of PMZ and PMZSO in muscle, liver, and kidney spiked samples were 0.05 μg/kg and 0.1 μg/kg, respectively, while those of Nor1PMZ were 0.1 μg/kg and 0.5 μg/kg, respectively. The LOD and LOQ of analytes in fat spiked sample were 0.05 μg/kg and 0.1 μg/kg, respectively. The sensitivity of this method reaches or exceeds its presented in previous reports. The analytes PMZ and PMZSO showed good linearity within the range of 0.1 μg/kg to 50 μg/kg, while Nor1PMZ showed good linearity within the range of 0.5 μg/kg to 50 μg/kg, with correlation coefficients (r) greater than 0.99. The recoveries of the target compounds in the sam-ples were between 81.73% and 107.17%, and the precision ranged from 1.78% to 10.47%. This study developed for the first time an HPLC-MS/MS method for the determination of PMZ and its metabolites PMZSO and Nor1PMZ in four swine edible tissues, comprehensively covered target tissues of monitoring object, which is applicable for monitoring veterinary drug residues in ani-mal-derived foods and ensuring food safety.

The pathogenesis of porcine circovirus type 2b (PCV2b) and swine influenza A virus (SwIV) during co-infection in swine respiratory cells is poorly understood. To elucidate the impact of PCV2b/SwIV co-infection, newborn porcine tracheal epithelial cells (NPTr) and immortalized porcine alveolar macrophages (iPAM 3D4/21) were co-infected with PCV2b and SwIV (H1N1 or H3N2 genotype). Viral replication, cell viability and cytokine mRNA expression were determined and compared between single-infected and co-infected cells. Finally, 3’mRNA sequencing was performed to identify the modulation of gene expression and cellular pathways in co-infected cells. It was found that PCV2b significantly decreased and improved SwIV replication, in co-infected NPTr and iPAM 3D4/21 cells respectively, compared to single infected cells. Interestingly, PCV2b/SwIV co-infection synergistically up-regulated IFN expression in NPTr cells whereas in iPAM 3D4/21 cells, PCV2b impaired the SwIV IFN induced response, both correlating with SwIV replication modulation. RNA-sequencing analyses revealed that the modulation of gene expression and enriched cellular pathways during PCV2b/SwIV H1N1 co-infection is regulated in a cell type-dependent-manner. This study revealed different outcomes of PCV2b/SwIV co-infection in porcine epithelial cells and macrophages and provides new insights on porcine viral co-infections pathogenesis.

Rapid spread of African swine fever virus (ASFV), causing severe disease with often high fatality rates in Eurasian suids, prevails as a threat for pig populations and dependent industries world-wide. Although advancing scientific progress continually enhances our understanding of ASFV pathogenesis, alternative transmission routes for ASFV have yet to be assessed. Here, we demonstrate that ASFV can efficiently be transferred from infected boars to naïve recipient gilts through artificial insemination (AI). In modern pig production, semen from boar studs often supplies many sow herds. Thus, the infection of a boar stud presents the risk of rapidly and widely distributing ASFV within or between countries. Daily blood and semen collection from four boars after intramuscular inoculation with ASFV strain ‘Estonia 2014’ resulted in detection of ASFV genomes in the semen as early as 2 dpi, in blood at 1 dpi while semen quality remained largely unaffected. Ultimately, after insemination with extended semen, 7 of 14 gilts were ASFV positive by 7 days post insemination, and all gilts were ASFV positive by 35 days post insemi-nation. Twelve out of 13 gilts aborted at the onset of fever. A proportion of fetuses originating from the remaining gilt showed both abnormalities and replication of ASFV in fetal tissues. Thus, we present evidence for the efficient transmission of ASFV to gilts via AI and also to im-planted embryos. These results underline the critical role that boar semen could play in ASFV transmission.

African swine fever virus (ASFV) is the causative agent of African swine fever, an economically important disease of pigs, often with a high case fatality rate. ASFV has demonstrated low genetic diversity among isolates collected within Eurasia. To explore the influence of viral variants on clinical outcomes and infection-dynamics in pigs experimentally infected with ASFV, we have designed a deep sequencing strategy. Variant analysis revealed 3 distinct SNPs present in the virus inoculum (at 2.4%, 0.7%, and 13% frequency, respectively) that were maintained within all infected pigs (1-6 % frequency, and the latter at 16-21%). Several pigs displayed other unique SNPs at &lt;10% frequency. In addition, a deletion of 10487 bp (resulting in the complete loss of 21 genes) was present at nearly 100% frequency in the ASFV DNA from one pig at position 6362-16849. This deletion was also found to be present at low levels in the virus inoculum and in two other infected pigs. The current methodology can be used for the currently circulating Eurasian ASFVs and also adapted to other ASFV strains and genotypes. Comprehensive deep sequencing is critical for following ASFV molecular evolution, especially for identification of modifications that affect virus virulence.

African swine fever virus (ASFV) is a large double stranded DNA arbovirus that is highly contagious and seriously endangers the lives of domestic and wild pigs. In the past decade, African swine fever (ASF) has spread in many countries in the Caucasus, Russian Federation, Eastern Europe and Asia, causing significant losses to the pig industry. At present, there is a lack of effective vaccine and treatment for ASF. Therefore, the rapid and accurate detection is crucial for ASF prevention and control. In this study, we have developed a portable lateral flow strip (LFS) detection method mediated by recombinase polymerase amplification (RPA) and DNA enzyme (LwCas13a), which is performed at 37 ℃ and visualized by eyes without the need for complex instruments. This RPA-LwCas13a-LFS is based on the ASFV structural protein p17 gene (D117L), with a detection sensitivity up to 2 gene copies. This method is highly specific and has no cross reactivity to 7 other pig viruses. In the detection of 33 clinical samples, the p17 (D117L) RPA-LwCas13a-LFS had 100% coincidence with conventional quantitative PCR (qPCR). These findings demonstrate the potential of this simple, rapid, sensitive, and specific ASFV detection method for on-site ASFV detection.

The rapid spread of African swine fever virus (ASFV), causing severe and often lethal disease in domestic pigs and Eurasian wild boar, continues to be a threat for pig populations and dependent industries. Despite scientific achievements that deepen our understanding of ASFV pathogenesis, alternative transmission routes for ASFV remain to be elucidated. We previously demonstrated the efficient transmission of ASFV from infected boars to naïve recipient gilts via artificial insemination, thereby highlighting the importance of surveillance in boar semen prior to shipment. Since accurate and reliable detection of even low amounts of ASFV in boar semen is key for disease prevention and control, we established a suitable diagnostic pipeline to efficiently detect ASFV genome in boar semen. Here, we assessed sensitivity, specificity and overall performance of various routine nucleic acid extraction kits as well as qPCR protocols in detecting ASFV genome in blood and semen of boars. Feasibility of respective kits and methods for future use in boar studs was also considered. Variability in sensitivity mostly concerned samples with low to very low amounts of ASFV genome. Ultimately, we defined a well-suited pipeline to precisely detect ASFV genome in boar semen, as early as 2 days post ASFV infection.

Proper treatment of infectious air could potentially mitigate the spread of airborne viruses such as porcine reproductive and respiratory syndrome virus (PRRSV). The objective of this research is to test the effectiveness of ultraviolet (UV) in inactivating aerosolized PRRSV, specifically, four UV lamps, UV-A (365 nm, both fluorescent and LED-based), "excimer" UV-C (222 nm), and germicidal UV-C (254 nm), were tested. The two UV-C lamps effectively irradiated fast-moving PRRSV aerosols with short treatment times (<2 s). One-stage and two-stage UV inactivation models estimated the UV doses needed for target percentage (%) reductions on PRRSV titer. UV-C (254 nm) dose needed for 3-log (99.9%) reduction was 0.521 and 0.0943 mJ/cm2, respectively, based on one-stage and two-stage models. An order of magnitude lower UV-C (222 nm) doses were needed for a 3-log reduction, i.e., 0.0882 and 0.048 mJ/cm2, based on one-stage and two-stage models, respectively. However, the cost of 222-nm excimer lamps is still economically prohibitive for scaling-up trials. The UV-A (365 nm) lamps could not reduce PRRSV titers for tested doses up to 4.11 mJ/cm2. Pilot-scale or farm-scale testing of UV-C on PRRSV aerosols simulating barn ventilation rates are recommended based on its effectiveness and reasonable costs comparable to HEPA filtration.

Porcine reproductive and respiratory syndrome virus (PRRSV) infections cause significant economic losses to swine producers every year. Aerosols containing infectious PRRSV are an important route of transmission, and proper treatment of air could mitigate the airborne spread of the virus within and between barns. Previous bioaerosol studies focused on the microbiology of PRRSV aerosols; thus, the current study addressed the engineering aspects of virus aerosolization and collection. Specific objectives were to (1) build and test a virus aerosolization system, (2) achieve a uniform and repeatable aerosol generation and collection throughout all replicates, (3) identify and minimize sources of variation, (4) verify that the collection system (impingers) performed similarly. The system for virus aerosolization was built and tested (Obj. 1). The uniform airflow distribution was confirmed using a physical tracer (<12% relative standard deviation) for all treatments and sound engineering control of flow rates (Obj. 2). Theoretical uncertainty analyses and mass balance calculations showed <3% loss of air mass flow rate between the inlet and outlet (Obj. 3). A comparison of TCID₅₀ values among impinger fluids showed no statistical difference between any two of the three trials (p-value = 0.148, 0.357, 0.846) (Obj. 4). These results showed that the readiness of the system for research on virus aerosolization and treatment (e.g., by ultraviolet light), as well as its potential use for research on other types of airborne pathogens and their mitigation on a laboratory scale.

African swine fever (ASF) is a notifiable viral disease of domestic and wild suids. Despite intensive re-search efforts, the pathogenesis of the disease is still far from being understood. Analysis of biomarkers in different body fluids may supplement traditional pathogenesis studies. As reliable protocols are often es-tablished in laboratories with lower biosafety, reliable inactivation of samples is crucial. The objective of this study was to find a procedure that inactivates the virus while preserving the biomarkers for down-stream analyses. To this means, three different inactivation protocols were employed, namely Tergitol-type NP-40 (NP-40) and polyoxyethylene-p-t-octylphenol (Triton X-100), respectively, and one with 95 °C heating. It could be demonstrated that all samples treated with 0.5% (v/v) concentration of both deter-gents showed absence of virus infectivity. The same was true for heated samples. However, heated serum was not suitable for analyses. Next, the treatment impact on biomarker readouts was assessed. While all protocols had an impact on the detection of biomarkers, correlation was retained. Especially NP-40 could be the desired detergent for more accurate measurements while achieving efficient virus inactivation. Based on these studies, samples can be reliably inactivated for most biomarker analyses and thus broader interdisciplinary cooperation is possible.

African swine fever virus (ASFV) continues to spread globally causing severe economic losses to the pig farmers. Vietnam licensed two live attenuated vaccines based on the ASFV strains ASFV-G-ΔI177L and ASFV-G-ΔMGF, to control the ongoing ASF outbreaks. In 2023, newly emerging highly virulent recombinant ASFV viruses containing genetic elements from both ASFV p72 genotype I and II (rASFV I/II) strains were reported from Northern Vietnam. This study evaluated if the two vaccine strains were able to protect the pigs against the emerging rASFV I/II strain VNUA/rASFV/TN1/23. Pigs vaccinated with ASFV-G-ΔMGF or ASFV-G-ΔI177L, when challenged with rASFV I/II, succumbed to the infection, or developed signs of chronic ASF. The findings from this study show that both vaccine strains that are licensed and used in Vietnam, are unlikely to protect pigs from the emerging highly virulent rASFV I/II strains. This complicates the ongoing efforts to control ASF in Asia and globally and emphasizes the urgent need for a novel vaccine that can effectively protect pigs from the rASFV I/II strains.

The spillover of a virus from an animal reservoir to humans requires both molecular and ecological risk factors to align. While extensive research both before and after the emergence of SARS-CoV-2 in 2019 implicates horseshoe bats as the significant animal reservoir for the new human coronavirus, it remains unclear why it emerged at this time. One massive disruption to animal-human contacts in 2019 is linked to the on-going African swine fever virus (ASFV) pandemic. Pork is the major meat source in the Chinese diet. We hypothesize that the dramatic shortage of pork following large-scale culling and restrictions of pig movement (resulting in marked price increases) led to alternative sources of meat and unusual animal and meat movements nationwide, e.g., involving wildlife, and thus greatly increased opportunities for human-sarbecovirus contacts. Pork prices were particularly high in southern provinces (Guangdong, Guangxi, Fujian, Jiangxi, Hunan, and Hubei), where wildlife is farmed and more frequently consumed. Major wildlife farming provinces are spread from Northern to Southern China, which overlaps with horseshoe bat host ranges, potential hosts of the proximal SARS-CoV-2 ancestor, and wildlife sourcing provinces of Wuhan Huanan market and possibly other markets. Trading of SARS-CoV-2 susceptible wildlife in these markets, such as minks, raccoon dogs, foxes and palm civets in Wuhan markets, could have increased the risk of SARS-CoV-2 from an intermediary host. Moreover, large quantities of animals raised for fur could have entered the human food chain undetected and significantly increased risks of animal-human contact. Performing retrospective testing of stored susceptible animals and their meat sold before December 2019 may be helpful in the next stage of tracing the animal origin of SARS-CoV-2 as spillover events are more likely to have taken place in 2019 when China was experiencing the worst effects of the ASFV pandemic.

This study reports the use of Nanotrap® Microbiome A Particles (NMAPs) to capture and concentrate viruses from diluted samples/specimens to improve their recovery and sensitivity of detection by real-time PCR/RT-PCR (qPCR/RT-qPCR). Five highly infectious animal disease viruses including goat pox virus (GTPV), sheep pox virus (SPPV), lumpy skin disease virus (LSDV), peste des petits ruminants virus (PPRV), and African swine fever virus (ASFV) were used in this study. After capture, the viruses remained viable and recoverable by virus isolation (VI) using susceptible cell lines. To assess efficacy of recovery, the viruses were serially diluted in phosphate buffered saline (PBS) or Eagle’s Minimum Essential Medium (EMEM) and then subjected to virus capture using NMAPs. The NMAPs and the captured viruses were clarified on a magnetic stand, reconstituted in PBS or EMEM, and analyzed separately by VI and virus-specific qPCR/RT-qPCR. The results of the qPCR/RT-qPCR showed up to 100-fold increase in the sensitivity of detection of the viruses following virus capture compared to the untreated viruses from the same dilutions. NMAPs were used to capture and concentrate viruses from clinical specimens to determine diagnostic sensitivity (DSe) that were comparable (100%) to the untreated specimens. NMAPs were also used to capture spiked viruses from EDTA whole blood (EWB). Virus capture from EWB was partially blocked, most likely by hemoglobin (HMB), which also binds NMAPs and outcompetes the viruses. The effect of HMB could be removed by either dilution (in PBS) or using HemogloBind™ (Biotech Support Group; Monmouth Junction, NJ) that specifically binds and precipitates HMB. Enhanced recovery and detection of viruses using NMAPs can be applicable to other highly pathogenic animal viruses of agricultural importance.

It is important to be able to detect and differentiate between distinct porcine enteric coronaviruses that can cause similar diseases. However, the existence of naturally occurring recombinant coronaviruses such as swine enteric coronavirus (SeCoV) can give misleading results with currently used diagnostic methods. Therefore, we have developed and validated three duplex real-time quantitative RT-PCR assays for the simultaneous detection of, and differentiation between, porcine epidemic diarrhea virus (PEDV) and SeCoV. Transmissible gastroenteritis virus (TGEV) is also detected by two out of these three assays. In addition, a novel triplex assay was set up that was able to detect and differentiate between these alphacoronaviruses and the porcine deltacoronavirus (PDCoV). The validated assays have low limits of detection, close to 100% efficiency and were able to correctly identify the presence of PEDV and SeCoV in 55 field samples, while 20 samples of other pathogens did not give a positive result. Implementing one or more of these multiplex assays into the routine diagnostic surveillance for PEDV will ensure that the presence of SeCoV, TGEV and PDCoV will not go unnoticed.

At the end of 2019, while the world was being hit by the COVID-19 virus and, consequently, was living a global health crisis, many other pandemics were putting humankind in danger. The role of social media is of paramount importance in these kinds of contexts since they help health systems to cope with emergencies by contributing to conducting some activities such as the identification of public concerns, the detection of infections’ symptoms, and the traceability of the virus diffusion. In this paper, we have analyzed comments on events related to cholera, ebola, HIV/AIDS, influenza, malaria, Spanish influenza, swine flu, tuberculosis, typhus, yellow fever, and zika, collecting 369,472 tweets from the 3rd of March to the 15th of September, 2022. Our analysis has started with the collection of comments composed of unstructured texts on which we have applied natural language processing solutions. Afterward, we have employed topic modelling and sentiment analysis techniques to obtain a collection of people’s concerns and attitudes toward these pandemics. According to our findings, people's discussions were mostly about malaria, influenza, and tuberculosis and the focus was on the diseases themselves. As regards emotions, the most popular were fear, trust, and disgust where trust is mainly regarding HIV/AIDS tweets.

Maternal-derived immunity is a critical component for survival and success of offspring in pigs to protect from circulating pathogens like Type 2 Porcine Reproductive and Respiratory Syndrome Virus (PRRSV-2). The purpose of this study was to investigate the transfer of anti-PRRSV immunity to piglets from gilts that received modified-live virus (MLV) alone (TRT 0), or in combination with one of two autogenous inactivated vaccines (AIVs, TRT 1+2). Piglets from these gilts were challenged with the autogenous PRRSV-2 strain at two weeks of age and their adaptive immune response (IR) was evaluated until 4 weeks post inoculation (wpi). The systemic humoral and cellular IR was analyzed in the pre-farrow gilts, and in piglets, pre-inoculation, and at 2- and 4-wpi. Both AIVs partially protected the piglets with reduced lung pathology and increased weight gain; TRT 1 also lowered piglet viremia best explained by the AIV-induced production of neutralizing antibodies in gilts and their transfer to the piglets. In piglets, pre-inoculation, the main systemic IFN-γ producers were CD21α+ B cells. From 0 to 4 wpi, the role of these B cells declined and CD4 T cells became the primary systemic IFN-γ producers. In lungs, CD8 T cells were the primary and CD4 T cells the secondary IFN-γ producers including a novel subset of porcine CD8α-CCR7- CD4 T cells, potentially terminally differentiated CD4 TEMRA cells. In summary, this study demonstrates that maternal AIV vaccination can improve protection of pre-weaning piglets against PRRSV-2; it shows the importance of transferring neutralizing antibodies to piglets; and it introduces two novel immune cell subsets in pigs – IFN-γ producing CD21α+ B cells and CD8α-CCR7- CD4 T cells.