Control and eradication of brucellosis is an objective for Veterinary and Health Authorities in several countries worldwide. Efficient surveillance programs are crucial for detecting and managing outbreaks, and epidemiological investigations may take advantage from the presence of standardized and efficient molecular typing techniques and analytical tools that enable public health laboratories to identify the origin of an outbreak. The aim of this study was to sequence Brucella spp. strains isolated in Iraq from different ruminant species, to verify whether there was a spatial or temporal clustering and, above all, shedding light on how these Iraqi isolates are positioned in the phylogenetic context of Brucella spp. The isolates under study (N=35) were from abortion, milk, placenta and fetal membrane of sheep, cattle and buffalo. Samples were genotyped using different techniques, namely MLVA-16, Whole Genome Sequencing, MLST and cgMLST. The findings of this study suggests that even relatively small investigations can provide sufficient evidence to justify improvements in existing control strategies. The application of different control measures in different territories, based on molecular epidemiological studies, would increase the chances to maximize public health benefits and minimize the spread of infection to areas free from the disease or with lower prevalence.

Unidentified abortion, of which leptospirosis, brucellosis and ovine enzootic abortion are important factors, is the main cause of disease spread between animals and humans in all agricultural systems in most developing countries. Although there are well-defined risk factors for these diseases, these characteristics do not represent the prevalence of the disease in different regions. This study predicts the unidentified abortion burden from multi-microorganisms in ewes based on artificial neural networks approach and GLM. Methods: A two-stage cluster survey design was conducted to estimate the seroprevalence of abortifacient microorganisms, and to identify putative factors of infectious abortion. Results: Overall seroprevalence of Brucella was 70.7%, while Leptospira spp. was 55.2%, C. abortus 21.9% and B. ovis 7.4%. Serological detection with the 4 abortion-causing microorganisms was determined only in 0.87% of sheep sampled. The best GLM is integrated by serological detection of serovar Hardjo and Brucella ovis in animals of the slopes with elevation between 2600 to 2800 meters above sea level from municipality of Xalatlaco, as well water supply, sheep pen built with materials of metal grids and untreated wood, with dirt and concrete floors, and bed of straw that remained independently associated with infectious abortion in ewes. Approximately 80% of those respondents did not wear gloves or masks to prevent the transmission of the abortifacient zoonotic microorganisms. Conclusions: Sensitizing stakeholders on good agricultural practices could improve public health surveillance. Further studies on the effect of animal-human transmission in such a setting are worthwhile to further support the One Health initiative.

In livestock, brucellosis is mainly an asymptomatic disease except for abortion, therefore two serological tests are used for diagnosis as no single test is suitable. Abattoir samples enable a combination of culture, molecular and serological tests to detect brucellosis. This study assessed Brucella specific PCR (ITS-PCR) to detect brucellosis and to conduct molecular characterization of Brucella spp. isolated from PCR-positive livestock (n=565) slaughtered at abattoirs and the appropriate sample tissue(s). ITS-PCR detected Brucella DNA in 33.6% cattle, 14.5% sheep and 4.7% pig tissues. Impure Brucella cultures from PCR-positive tissues were 43.6% (44/94) cattle, 51.7% (15/29) sheep, and 50% (2/4) pigs with predominantly B. abortus identification with AMOS-PCR and low isolation of mixed B. abortus and B. melitensis in all species. In cattle 33% of isolates were from lymph nodes while in sheep 38.0% were from liver and kidney and only from tonsils in pigs (2/4). Brucella infections identified with AMOS-PCR were present in seropositive and mainly seronegative (75.6-100%) livestock with the potential to cause brucellosis during pregnancy or breeding. This study demonstrated the value of the polyphasic approach, especially with chronic infections and the potential risk of these asymptomatic animals.

Brucellosis is an important infectious disease caused by bacteria of the genus Brucella. In the northeast region of Portugal, brucellosis is endemic in small ruminants, and there are also human cases. However, the epidemiological role of the wild boar in the dynamics of this disease in this region is unknown. For this reason, a total of 332 blood samples were collected from wild boar hunted in thirty-six hunting areas during the 2022/2023 hunting season. All were taken by the hunters for private consumption, with no evisceration and examination in the field. Serum sam-ples were tested by i-ELISA. It was observed that 88 wild boars were exposed to Brucella spp., pointing to a seroprevalence of 26.5% (95% CI: 21.8% – 31.3%). This high prevalence underlines the importance that wild boar may have in the dynamics of this disease in the region and its po-tential transmission to other animals and to human, for example, during the handling of carcasses. Increased awareness and knowledge of brucellosis in wild boar is essential for the implementation of effective practices and habits and, consequently, for the control and prevention of this important zoonosis.

This investigation aimed to knowledge of the presence of Brucella spp .in the samples from Babylon province and the relationship with environmental factors. The seasonal collection of animal soil samples for various villages in the middle, south, and north of Babylon province, Hilla City, Iraq, with physicochemical parameters (air and soil temperature, pH, EC, TDS, salinity, TOC), as well as heavy metals (Fe, Cu, Cd, and Pb). The results showed the presence of Brucella species in some of soil samples and its percentage varies according to the four seasons, where the highest percentage of Brucella spp. was in summer season and the lowest in winter season and these percentages were related to seasonal environmental indicators. After sending the positive bacterial samples isolated from the soil for sequencing analysis, it was found that the bacterial samples isolated from the soil have new strains recorded in Genbank as following: five strains for Brucella intermedia and one strain for Brucella rhizosphaerae, Also, phylogenetic analysis was for the drawing of phylogenetic tree and the determination of phylogenetic relationship for Knowledge the degree of convergence between species and their source. Through our research, we concluded that proportion of Brucella spp. presence in summer was high and also environmental factors which included (Air and Soil temperature, pH, EC, TDS and Salinity) were high, except TOC was lower in summer season whereas heavy metals for Cu, Fe, Cd were lower in summer except Pb was high. This indicated that differences of seasons affect to Brucella ratio in soil. The findings of sequencing for the bacterial samples from soil demonstrated that B. intermedia was found in both the spring and fall seasons while B.rhizosphaerae found in the spring season , These types of Brucella had genetic variations. Also concluded that during phylogenetic analysis of the 16S rRNA sequences indicated that B.intermedia was nearest to Pakistan and other strain of B.intermedia was nearest to Russia while B.rhizosphaerae was nearest to Ukraine and Pakistan.

Catalase, an antioxidant enzyme widely produced in mammalian cells and bacteria, is crucial in mitigating oxidative stress in a hostile environment. This function enhances the intracellular survivability of various intracellular growth pathogens, including Brucella abortus. In this study, to determine whether the inhibition of catalase can inhibit the intracellular growth of Brucella (B.) abortus, we employed 3-amino-1,2,4-triazole (3-AT), a catalase inhibitor, in both RAW 264.7 macrophage cells and an ICR mouse model during Brucella infection. The intracellular growth assay indicated that 3-AT exerts growth-inhibitory effects on B. abortus within macrophages. Aside from that, it contributes to the accumulation of reactive oxygen species and the formation of nitric oxide. Notably, 3-AT diminishes the activation of the nucleus transcription factor (NF-κB) and modulates cytokine secretion within infected cells. In our mouse model, the administration of 3-AT reduced B. abortus proliferation within the spleen and liver of infected mice. This reduction was accompanied by a diminished immune response to infection, as indicated by lowered levels of TNF-α, IL-6, IL-10, and altered CD4+/CD8+ T-cell ratio. These results suggest the protective and immunomodulatory effects of 3-AT treatment against Brucella infection.