Toxoplasma gondii in slaughtered sheep: a study of parasite prevalence, isolation, genotyping, virulence, and potential health risks to butchers in India.

PCDD/Fs in food products produced near a contaminated former sawmill - concentrations, congener profiles and risk assessment.

Mineral content plays a pivotal role in determining the quality of meat, which consumers prioritizewhen seeking healthy and high-quality lamb. Enhancing the genetic makeup of mineral content istherefore crucial to pique consumers’ interest in lamb meat. Among the potential genes that could beused as markers for the nutritional quality, including the mineral content, of Indonesian lamb meat, thecytochrome P450, family 2, subfamily A, polypeptide 6 (CYP2A6) gene stand out. The main objectiveof this study was to identify the CYP2A6 gene polymorphism at SNP g.49170107 G>T and explore itsassociation with mineral content in Indonesian sheep. For this purpose, 95 sheep samples were analyzed,consisting of 81 Javanese thin-tail sheep (JTTS) and 14 Jonggol sheep (JS). The researchers investigatedthe polymorphism through the PCR-RFLP (Polymerase Chain Reaction - Restriction Fragment LengthPolymorphism) method. Subsequently, they analyzed the association between the CYP2A6 gene andmineral content in lamb meat using the GLM (General Linear Model) method. The study revealedthat the CYP2A6 gene polymorphism was present in JTTS, while it remained monomorphic in JS.The identified genotypes in the polymorphism were GG and GT. Notably, the CYP2A6 gene exhibiteda significant association (P<0.05) with Fe content. Specifically, the GT genotype displayed higher Fecontent compared to the GG genotype. These findings suggest that the CYP2A6 gene holds promise as apotential candidate for identifying mineral content in lamb meat.

Shiga toxin-producing Escherichia coli (STEC) is a zoonotic pathogen of public health concern, requiring a One Health approach to clarify its transmission and distribution. However, its prevalence and genomic characteristics in livestock and companion animals remain underexplored in low-income countries. We investigated prevalence and genomic features of STEC in animals in western Ghana, representing the first genomic report of STEC in Ghana. Fecal samples (97) were collected from goats (n = 33), sheep (n = 33), dogs (n = 30), and a cat (n = 1), with STEC detected in 12.1% of goats and sheep samples. Whole-genome sequencing identified serotypes O38:H26, O43:H2, and O157:H7. stx1c and stx2b genes were detected in O38:H26 and O43:H2, whereas stx2c and key virulence genes (chuA, eae, esp, nle, tir, and toxB) were exclusively found in O157:H7. Phylogenetic analysis revealed that O38:H26 isolates form a cluster closely related to clinical strains from the UK. O43:H2 isolates exhibited diverse stx profiles, linking animal, environmental, and clinical strains from North America and the UK. O157:H7 isolates were genetically similar to European clinical and food-derived strains, suggesting that goats and sheep are important STEC reservoirs in Ghana, offering data for public health risk assessment and effective One Health-based control strategies.

The aim of this work was to study the associations of BMPR-IB gene polymorphism with the concentration of macroelements (Na, Mg, K) in the blood serum of Romanov sheep. The level of macroelements in animal organs and tissues is one of the important indicators for a comprehensive characterization of the interior. The BMPR-IB gene, which encodes a receptor for bone morphogenetic proteins, plays a key role in reproductive function; however, its influence on mineral metabolism has not been sufficiently studied. The study used a sample of sheep (n = 58) kept under the same conditions and on a standard diet. Using PCR-RFLP and non-parametric statistics, statistically significant differences in sodium concentration between the ++ and M+ genotypes were revealed (Kruskal-Wallis test, p = 0.00192; Dunn’s post-hoc test, p = 0.0014). It was found that heterozygous (M+) animals had a significantly higher sodium concentration by 43 % (136.30 vs. 95.15 mmol/L) compared to wild-type homozygotes (++), with a large effect size (r = 0.54). The statistical significance remained after correction for multiple comparisons (p_FDR = 0.0058). No significant associations with the genotype were found for magnesium and potassium. The obtained data indicate a pleiotropic effect of the BMPR-IB gene on mineral metabolism and reveal the potential of using the M+ genotype as a molecular marker in breeding programs for optimizing the metabolic status of sheep. The identified association suggests a possible involvement of this gene in the regulation of kidney function and water-salt homeostasis, opening new directions for fundamental research in animal physiology. The results of the work emphasize the importance of comprehensive study of genetic markers for developing effective breeding strategies for farm animals.

Brucellosis is listed as a priority disease in low-income countries like Guinea, facing challenges in logistics, equipment, competence, and cost limitations for diagnosis. Serological diagnosis is mainly performed by the Rose Bengal agglutination test (RBT) in the veterinary sector. We have compared its discriminative capacity with more sophisticated and expensive serological tests, such as multi-species or species-specific ELISA kits and Complement Fixation test (CFT). A panel of 554 serum samples of pigs, goats, sheep, and cattle collected throughout Guinea from 2017 to 2019 where tested by RTB and ELISA tests in parallel at the Institut Pasteur de Guinée (Conakry) and the Brucellosis WOAH/EU Reference Laboratory of the French Agency for Food, Environmental and Occupational Health & Safety (Maisons-Alfort, France). ELISAs performed equally across laboratories (Kappa =0.867-0.958); RBT and ELISA showed 94-95% concordance. The CFT value of positive cattle samples also logically followed the RBT scores CONCLUSIONS/SIGNIFICANCE: In low-income countries like Guinea, the less expensive RBT can be regarded as a convenient routine Brucella diagnosis tool, assuming a solid experience of the operator following standard operating protocols and regular proficiency tests. As WOAH recommends confirmatory methods, the multispecies ELISA kit appears as a good candidate for conveniently trained and equipped laboratories.

Peste des petits ruminants (PPR) is a highly contagious viral disease prevalent in sheep and goats, and causes significant economic losses. The study was conducted in 2024 in Punjab Province, Pakistan, to estimate the seroprevalence of the PPR virus (PPRV) and to analyze animal‐level risk factors in unvaccinated small ruminants. Over a 12‐month period, multistage random sampling provided 722 serum samples of sheep and goats aged 6 months or older, collected across six districts. The anti‐PPRV antibodies were detected using competitive enzyme‐linked immunosorbent assay (cELISA), and species, age, sex, breed, parity, lactation status, pregnancy status, body condition score (BCS), and reproductive history were analyzed using univariable and multivariable logistic regression analyses. The overall seroprevalence rate was 79.77%, but significantly higher in goats (90.27%) than in sheep (68.75% and p < 0.0001). The results demonstrated district wise disparity, with variation in seroprevalence between districts: 52.05% (Okara) and 100% (Nankana). An increased likelihood of seropositivity was found to be associated with male sex, some breeds (Makhi Cheeni and Beetal), certain species‐district interactions, and age‐sex interactions. These data confirm the high endemicity of PPRV in Punjab and justify the targeted vaccination and surveillance in high‐risk areas and among susceptible animal populations.

This study was aimed to identify the genetic polymorphism of the Secreted phosphoprotein 1 (SPP1) gene in a sample of Awassi sheep consisting of 48 ewes and its 62 newborns. This study lasted for six months, The results revealed that there were two variations (2 SNPs) in the Promoter Exon 3 of gene SPP1, the first variance SNP1 the position was (SNP1 A &gt; G: P/1212/(84 bp), AG. The second variance Snp2, G &gt; A: P/1207/ (92 bp), There were two genetic components of this gene, GG and GA, at a percentage rate of 60.42 and 39.58%, respectively, and there were high-morale (P≤0.01) differences in the birth weight of the ewe, The GA hybrid ewe outperformed the G-pure terrestrial genetic structure. This study concludes that the mutant GG genetic structure does not appear in the SNP1 and AA does not appear in the SNP2 of the gene's studied region. There is a correlation between the heterogeneity obtained from the study of SPP1 gene and the production of daily milk and some liver enzymes in passive sheep that can be exploited in election and improvement programmes.

Objective: Climate change lies at the core of the multifaceted global crises confronting humanity today. Sheep breeding plays a crucial role in ensuring the sustainability of global food security. Heat stress resulting from the increasing rise in global temperatures necessitates the genetic improvement of sheep breeds in terms of the genes that determine their resistance to this stress. The present study aimed to screen single-nucleotide polymorphisms (SNPs) located in the eighth exon of the HSPA8 gene, which is one of the important heat shock proteins, in Artlı and Çepni sheep populations using the Sanger sequencing method. Material and Methods: The eighth exon region of the HSPA8 gene was sequenced, and SNPs within the obtained sequences were identified using the MEGA X software. The potential effects of the detected mutations on the protein structure were modeled using the HOPE software. In addition, to assess the accuracy of the predicted protein structure, confidence scores (pLDDT) were obtained using the AlphaFold2 software. Results: Artlı sheep are traditionally known for their high resistance to low-temperature stress and harsh environmental conditions. In this study, a C&gt;T substitution at the 210th nucleotide position of the 8th exon of the HSPA8 gene was detected in 50% of the analyzed Artlı sheep samples, resulting in the replacement of proline with leucine at amino acid position 605. Furthermore, 40% of the Artlı genotypes exhibited a G&gt;A substitution at the 46th nucleotide position of the same exon, which was found to be heterozygous and caused a silent mutation at amino acid position 550. In contrast, all analyzed samples of the Çepni breed were found to be monomorphic with respect to the examined exon region. Conclusion: The genetic basis of adaptation to climate change is shaped not only by mutations but also by other genetic processes, such as epistatic interactions between genes and epigenetic regulatory mechanisms. Rising temperatures resulting from global climate change have led to a decline in livestock productivity. Sustainable sheep breeding depends on evaluating local breeds, such as Artlı and Çepni, as potential breeding materials. Therefore, there is a need to conduct comprehensive genetic studies on these unique genotypes and to strengthen efforts aimed at conserving local breeds.

Mycoplasma capricolum subsp. Capripneumoniae (Mccp) is the causative agent of contagious caprine plueropneumonia (CCPP). In many countries CCPP is diagnosed by clinical signs and serological methods only. Mccp is known to be difficult to culture in the laboratories due to the fastidious nature of the organism. In the Sudan, there is need for further studies on sheep and goat pneumonia. The aim of this study was to determine the frequency of Mccp in ovine and caprine pneumonic lungs and to study the histopathology of the affected lungs. Tissue samples were obtained from condemned lungs of slaughtered sheep and goats and from clinically ill goats in Omdurman, Khartoum State, Sudan. Bacteriological culture and PCR methods were used to detect Mccp. Histopathological evaluation was also performed. Mccp DNA was identified in 4 goat and one sheep sample using PCR, while only one of them- goat lung sample- was identified using bacteriologic culture lung samples. None of the detected microscopic lung lesion could be considered specific for Mccp infection. This study documents for the first time, the presence of Mycoplasma capricolum subsp. capripneumoniae (Mccp) DNA in pneumonic sheep lung in Sudan suggesting that sheep may act as potential carriers of CCPP and/or clinically affected animal species. This highlights the need for further investigation into the epidemiological role of sheep in the transmission of CCPP. Although PCR is more costly than traditional bacteriological culture, it proves to be a valuable tool for CCPP surveillance due to its higher sensitivity and specificity. Int. J. Appl. Sci. Biotechnol. Vol 13(4): 146-152.

Background and objective: Echinococcus granulosus can cause hydatidosis, or echinococcosis, in both herbivores and omnivores. Hydatid cysts are the larval stage of this tapeworm. As a zoonotic illness, it affects people all over the world. Devastating effects on human and animal health as well as massive economic losses, disproportionately felt in the agriculture sector, characterize this disease. Genomic segments from cox1 and nad1 genes have allowed for the identification of distinct genotypes of Echinococcus granulosus sensu stricto (s.s.) in both humans and domesticated animals. The current investigation aimed to distinguish between the G1/G3 genotypes of Echinococcus granulosus (s.s.) in cattle via the nad5 gene fragment. Methods: The present study was conducted in Erbil city. Sheep, goat, and cattle hydatid cyst samples (36 isolates) were collected from the Erbil slaughterhouse. In addition, 11 fresh human hydatid cyst samples were taken from patients with hydatid cysts who underwent surgical operations in Rizgary Teaching Hospital. The hydatid cysts were subjected to examination for fertility and viability in addition to DNA extraction for genotyping based on the Nad5 gene. Results: Out of 42 samples, 15 samples were sequenced, and all samples belonged to the G1 genotype (sheep strain). DNA sequencing of hydatid cysts from both animal and human sources showed mutations in various positions of the Nad5 gene, consistent with the dominant G1 genotype. In human samples, several variations overlap with other hosts, but unique ones include human 11,12 glycine/alanine at 142 arginine/glutamine, glycine/threonine at 303 valine/phenylalanine, T/A at 334 valine/aspartic acid, and T/C at 361 arginine/histidine, which result in 99% identity to the G1 strain. Conclusion: Humans and animals in Erbil most commonly harbor the G1 genotype of E. granulosuss. s.s. according to the study. The findings provided additional evidence that the Nad5 gene accurately distinguished between humans and cattle, sheep, goats, and E.granulosus G1/G3 isolates.

Background: Leptospirosis is a zoonotic disease, common in tropical and subtropical regions, caused by pathogenic species of Leptospira. Objectives: This study aimed to isolate and detect Leptospira in animal samples in Amol, northern Iran. Methods: A total of 118 samples were collected from rodent bladder urine and livestock midstream urine during 2023. Culture, microscopy, and molecular analyses were conducted to identify Leptospira interrogans. Cultures were incubated in Ellinghausen McCullough Johnson and Harris (EMJH) media and monitored weekly via dark field microscopy. DNA extraction was followed by conventional and real-time polymerase chain reaction (PCR) targeting the lipL32 gene. Multi locus sequence typing (MLST) was also performed on selected isolates. Results: Results showed that seven (5.93%) samples (including six rodent and one sheep samples) were culture-positive, all confirmed by real-time and traditional PCR. Real-time PCR identified 14 (11.86%) samples as positive for L. interrogans including 9/32 (28.1%), 3/44 (6.81%), and 2/42 (4.76%) rodent, sheep, and cattle samples, respectively. The results of the MLST sequence analysis of four culture-positive specimens were attributable to sequence type (ST) 330 (one sheep specimen) and ST 252 (three rodent specimens), both of which correspond to L. interrogans. Quantitative statistical analyses revealed noteworthy correlations between the types of samples and the presence of Leptospira (P &lt; 0.05). Conclusions: This investigation underscores the effectiveness of integrating molecular methodologies with conventional culture techniques for the surveillance of Leptospira in regions characterized by a heightened risk. The current study reports the isolation and characterization of Leptospira from animal urine samples, providing new insights into the presence of these bacteria in the study area.

ABSTRACT Milk is a complete food that plays a fundamental role in supporting bone health with high content of calcium and protein. Recent research highlights the significance of milk‐derived extracellular vesicles (mEVs) with emerging roles in modulating physiological processes, including bone remodeling. This study reports a comparative analysis of mEVs isolated from raw milk of three ruminant species: bovine, goat, and sheep. Extracellular vesicles (EVs) are characterized for size distribution, particle concentration, and protein composition. Functional relevance of each type of mEVs is evaluated on cell proliferation and osteogenic differentiation of murine fibroblasts model. Results reveal significant interspecies differences in both the structural and biological characteristics of mEVs. EVs derived from each milk type exhibit distinct effects on MC3T3‐E1 viability, supporting the idea that mEVs have a beneficial role in cell growth and proliferation and induce osteogenic differentiation. Additionally, the microbial DNA content carried within mEVs from each ruminant species harbors distinct and structured microbial DNA profiles. Goat mEVs are enriched in beneficial commensals, whereas sheep samples reflect consistent yet mixed communities, and bovine mEVs exhibit environmental signatures and greater variability. Overall, these findings highlight the species‐specific potential of mEVs, their compositional diversity, and potential functions of the microbial cargo. Furthermore, this study highlights mEVs as functional food components and opens new avenues for developing targeted dietary strategies leveraging mEVs to support personalised medicine in bone health and prevent or manage conditions such as osteoporosis.

Simple SummarySheep can be infected by parasites of the genus Sarcocystis, which form cysts in muscles and internal organs and may cause disease, economic losses, and animal welfare concerns. These protozoa have an obligate two-host life cycle, requiring intermediate hosts such as sheep and definitive hosts, including dogs or wild carnivores. Infections can be subclinical but may also cause weight loss, anemia, neurological signs, abortion, or death, depending on species and host susceptibility. In Portugal, where sheep farming is a key agricultural sector, data on the prevalence of Sarcocystis spp. in meat-producing animals were lacking. This study analyzed brain and blood samples from sheep and stool samples from shepherd dogs to detect the presence of the parasite using molecular methods. Sarcocystis sp. was identified in 4.9% of brain tissue samples, while no DNA was detected in blood or dog stool samples, representing the first molecular confirmation in Portuguese sheep destined for human consumption. Although dogs tested negative, other definitive hosts, including wild carnivores, may contribute to parasite circulation. These findings highlight the importance of understanding Sarcocystis distribution and transmission dynamics to support animal health, minimize economic losses, and guide effective control strategies in Portuguese sheep farming.Sarcocystis spp. are cyst-forming protozoan parasites with a global distribution that infect a wide range of domestic and wild animals, impacting both animal health and livestock productivity. In sheep, infections can cause clinical disease, reproductive losses, and economic damage, particularly when pathogenic species such as Sarcocystis tenella are involved. Grazing sheep, including breeds such as the Serra da Estrela from central Portugal, are at increased risk due to frequent contact with shepherd dogs, which serve as definitive hosts. Despite their significance, data on the occurrence and distribution of Sarcocystis spp. in Portuguese sheep remain limited. This study analyzed 179 samples collected in central Portugal during 2024, including 41 brain tissues and 88 blood samples from sheep, and 50 stool samples from shepherd dogs, using conventional PCR and bidirectional Sanger sequencing. Sarcocystis sp. closely related to S. tenella was detected exclusively in sheep brain tissue, with a prevalence of 4.9% (2/41; 95% CI: 0.60–16.53), while no parasite DNA was found in blood or dog samples. These results provide the first molecular confirmation of Sarcocystis spp. closely related to S. tenella in Portuguese sheep raised for human consumption and establish baseline data for future epidemiological surveillance and control strategies.

The Booroola fecundity (FecB) gene, a mutation in the bone morphogenetic protein receptor-1B (BMPR1B), is known to increase ovulation rate and litter size in sheep. Efficient and accurate, large-scale detection of this single nucleotide polymorphism (SNP) is critical for marker assisted introgression/selection programs for genetic improvement of prolificacy. This study aimed to develop and validate a robust, cost-effective, and high-throughput genotyping assay for detecting the FecB mutation across diverse sheep populations globally. A competitive allele-specific PCR (KASP) assay targeting the A746G nucleotide substitution in BMPR1B gene was designed and tested on a diverse panel of 224 individuals across 22 sheep breeds and validated against direct sequencing. The KASP genotyping results showed 100% concordance with sequence data, with clear fluorescence-based discrimination of homozygous wild-type Fec++ (AA), heterozygous FecB+ (AG), and homozygous mutant FecBB (GG) genotypes. The assay was compatible across three real-time PCR platforms, BioRad CFX96, Roche LightCycler 480 II, and ABI QuantStudio 6 demonstrating reproducible genotyping and cross-platform interoperability. Subsequently, the KASP assay was applied to a global screening of 1471 sheep from 47 breeds in 14 countries for further validation of the assay. The mutation was not observed in the sheep samples investigated from Africa, Europe, Latin America, and West Asia, whereas it was polymorphic or fixed in certain populations from South (India and Bangladesh) and Southeast Asia (Indonesia). Notably, native Bangladeshi sheep exhibited high frequencies of the FecB allele, with some populations (e.g., Bangladesh Central) nearing fixation, reflecting possible selection for high prolificacy. In conclusion, the validated KASP assay offers a powerful, scalable tool for the rapid genotyping of the FecB mutation, enabling efficient screening and incorporation of prolificacy traits in sheep breeding programs worldwide.

Congener-specific accumulation of PCDD/Fs, PCBs, and PBDEs in sheep (Ovies aries) liver and muscle.

Diarrhea in young ruminants is a global issue and causes significant economic losses worldwide. In addition to common pathogens like rotavirus, coronavirus, and astrovirus, new viruses can be identified through unbiased next-generation sequencing (NGS) techniques. Here, we report the initial identification of a hunnivirus from a one-month-old goat with diarrhea using shotgun metagenomic NGS. A complete hunnivirus genome was recovered. Phylogenetic tree analysis revealed that this goat hunnivirus was more closely related to cattle hunnivirus than to small ruminant hunnivirus strains, suggesting a prior cross-species transmission event. The genome was used to design primers/probes for the conserved 3Dpol RdRP gene for real-time RT-PCR to screen banked ruminant fecal samples. Screening of 144 ruminant fecal samples showed that 9 of 38 goat, 22 of 96 cattle, and 0 of 8 sheep samples were positive for hunnivirus. Sequencing of the 3Dpo region was performed on selected positive samples and revealed two lineages of hunnivirus circulating in North America. Our study highlights the importance of further investigation and monitoring of fecal samples using unbiased metagenomic tools to identify potential pathogens or co-infections in ruminants.

Cross-species virulence strategies of Mycobacterium avium subsp. paratuberculosis: Gene expression and infection progression in sheep and guanacos.

IntroductionIn the evolutionary context of sheep, the development of fat tails represents an adaptive survival mechanism in response to varying food availability. Despite food resource instability, sheep store energy by accumulating tail fat to survive periods of famine. This energy storage function remains present in domesticated sheep, serving as a key evolutionary reason for the formation of sheep tail fat.MethodsHere, we conducted whole-genome resequencing of 555 sheep samples (30 samples were newly sequenced and 525 were retrieved from published data) globally to investigate selection signatures associated with fat-tailed traits using Fixation Index (FST), Nucleotide diversity (π), cross-population composite likelihood ratio (XP-CLR), and runs of homozygosity (ROH) methods.Result and discussionOur examination of selection signatures in Fat-tailed and Thin-tailed Sheep Populations identified 32 candidate genes, with 6 genes (PDGFD, BMP2, GLIS1, LIPE, MSRB3, and TBX15) implicated in fat accumulation and lipid metabolism. Notably, 8 significant Gene Ontology terms (mesenchymal cell differentiation, positive regulation of ERK1 and ERK2 cascades, hormone metabolic process, nucleocytoplasmic transport, regulation of hormone levels, response to growth factor, regulation of canonical Wnt signaling pathway, and tissue morphogenesis) may play a role in fat deposition and tail fat development. These results will provide molecular targets for low-fat sheep breeding and enhance economic returns in sheep farming.ConclusionThis study will play a crucial role in environmental adaptation and product development, comprehensively driving the development of the sheep farming industry and enhancing economic benefits.

Crimean-Congo hemorrhagic fever virus (CCHFV) is a tick-borne zoonotic pathogen of growing public health concern in southeastern Europe. This study provides the first serological evidence of CCHFV circulation in Croatia, based on testing 1473 serum samples from farm and companion animals, including sheep, horses, cattle, goats, dogs, and cats. A total of 109 samples (7.4%) tested positive for CCHFV antibodies using a commercially available enzyme-linked immunosorbent assay (ELISA) kit. The highest seroprevalence was recorded in sheep (28.3%), followed by horses (4.3%) and a single cat (0.5%), with no antibodies detected in cattle, goats, or dogs. Almost all seropositive animals originated from coastal and subcoastal Croatia, where Hyalomma ticks are present. Only two seropositive cases were detected in continental areas. Sheep samples from several farms in Zadar County showed intra-farm seropositivity rates of up to 85.7%, suggesting localised virus circulation likely influenced by vector distribution and farm-level practices. No viral ribonucleic acid (RNA) was detected by quantitative reverse transcription polymerase chain reaction (qRT-PCR), consistent with the transient nature of viremia in most animal hosts. These findings confirm the silent circulation of CCHFV in Croatia and reinforce the need for targeted, regionally adapted surveillance strategies that integrate multiple hosts and support early warning systems aligned with the One Health concept.