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Background Water buffalo is one of the most important livestock species in the world. Two types of water buffalo exist: river buffalo ( Bubalus bubalis bubalis ) and swamp buffalo ( Bubalus bubalis carabanensis ). The buffalo genome has been recently sequenced, and thus a new 90 K single nucleotide polymorphism (SNP) bead chip has been developed. In this study, we investigated the genomic population structure and the level of inbreeding of 185 river and 153 swamp buffaloes using runs of homozygosity (ROH). Analyses were carried out jointly and separately for the two buffalo types. Results The SNP bead chip detected in swamp about one-third of the SNPs identified in the river type. In total, 18,116 ROH were detected in the combined data set (17,784 SNPs), and 16,251 of these were unique. ROH were present in both buffalo types mostly detected (~ 59%) in swamp buffalo. The number of ROH per animal was larger and genomic inbreeding was higher in swamp than river buffalo. In the separated datasets (46,891 and 17,690 SNPs for river and swamp type, respectively), 19,760 and 10,581 ROH were found in river and swamp, respectively. The genes that map to the ROH islands are associated with the adaptation to the environment, fitness traits and reproduction. Conclusions Analysis of ROH features in the genome of the two water buffalo types allowed their genomic characterization and highlighted differences between buffalo types and between breeds. A large ROH island on chromosome 2 was shared between river and swamp buffaloes and contained genes that are involved in environmental adaptation and reproduction.

This study investigates the potential of blending wheat germ oil (WGO) with local and imported Bubalus bubalis butter fats to enhance its physicochemical properties, oxidative stability, and sensory characteristics. Blends were prepared at ratios of 5%, 10%, and 15% WGO with butter fats and analyzed for fatty acid composition, oxidative stability, and sensory acceptability. Oxidative stability was assessed using peroxide value (PV), conjugated diene (CD), and thiobarbituric acid (TBA) tests. Results revealed that blending WGO with butter fats significantly enhanced oxidative stability. Local butterfat blends demonstrated superior oxidative stability, with a PV of 1.22 meq O2/kg compared to 1.65 meq O2/kg for imported butterfat blends after 30 days of storage—a 25% improvement. Sensory evaluation indicated that the 5% WGO and 95% butterfat blend achieved the highest scores in flavor, texture, and overall acceptability, with an overall score of 86.19%. In contrast, higher WGO ratios reduced sensory appeal. This study highlights the potential of WGO-butterfat blends to improve oxidative stability while maintaining sensory quality, with 5% WGO being the optimal blend ratio.   See corrigendum at: https://itjfs.com/index.php/ijfs/article/view/3603  

This study explored the effect of melatonin implantation on the fresh and cryopreserved semen quality of Iraqi buffalo bulls (Bubalus bubalis). Nine adult bulls were used and divided equally into three groups. The first group was left without treatment and regarded as control (T1), whereas the second (T2) and third (T3) groups were subcutaneously implanted with 72 and 90 mg of melatonin respectively on the left ear base and repeated one month post the first implantation. Semen was collected for 12 weeks and evaluated weekly for fresh, cooling, and post-cryopreserved (PC) protocols. The T3 group exhibited a higher (P≤0.05) sperm cell individual motility percentage, whereas, the T1 and T2 groups recorded better (P≤0.05) sperm acrosome integrity percentage for fresh semen. A lesser (P≤0.05) total sperm abnormality (TSA) percentage in the T2 and T3 groups and a greater (P≤0.05) sperm acrosome integrity (SAI) percentage in the T3 group were noticed during the cooling preservation. Moreover, the T3 group exhibited a lesser (P≤0.05) TSA percentage whereas, the T2 and T3 groups revealed a leaser (P≤0.05) SAI percentage during the PC period. Concomitantly, lower (P≤0.05) PC malondialdehyde concentration in seminal plasma was observed in the T2 and T3 groups than control group. In conclusion, the melatonin implantation (72 and 90 mg) ameliorated some fresh and PC semen attributes of the Iraqi buffalo bulls.

The wild water buffalo ( Bubalus arnee ) is one of the most endangered and least studied large bovid in the Indian subcontinent. India retains 90% of the estimated global population of >4000 individuals as two fragmented populations in Assam and Chhattisgarh, both threatened by habitat loss and degradation, hunting, disease from livestock, and hybridization with the domestic buffalos. Small, fragmented population size and potential hybridisation pressures from co-occurring domestic buffalos are the major conservation challenges. For the first time, we sequenced the 16,357 bp long mitogenome of three opportunistically collected wild water buffalo samples from Assam (n = 1) and Chhattishgarh (n = 2). The annotated sequence has a base composition of 26.4% T, 26.6% C, 33.1% A and 13.9% G depicting an AT-rich mitogenome composition, including 13 protein-coding genes (11,361 bp), 22 transfer RNA (tRNA) (1514 bp), two ribosomal genes (2525 bp), and a non-coding control region (928 bp). The gene order is conserved with other bovid species. Comparative mitogenome analyses showed both populations are genetically similar but significantly different from domestic buffalo. We also identified structural differences in seven tRNA secondary structures between both species. The genetic distance between wild buffalo and other bovids varied between 0.103 and 0.122. Multiple Bayesian phylogenetic trees showed that both wild and domestic water buffalo formed sister clades which were paraphyletic to other potentially sympatric species of genus Bos. This study provides baseline information on wild buffalo mitogenome for further research on phylogeny, phylogeography and hybrid assessment and help conserving this endangered species.

This case report describes for the first-time cases of severe gastroenteritis in water buffalo calves due to a new serovar of Salmonella enterica. The study was carried out on fecal matrix collected from live water buffalo calves that showed profuse diarrhea, severe dehydration and fever, exhibiting a systemic course. Culture and molecular investigations identified the pathogens isolated from intestinal contents as two Salmonella serovars, Salmonella enterica enterica O:35 and a new serovar of Salmonella enterica. The isolates showed multi-drug resistance. Timely diagnosis associated with a targeted antimicrobial treatment were found to be sufficient for the survival and recovery of the infected animals. Herd vaccines prepared from isolated pathogens were used to prevent further deaths of the calves.

Background The Asiatic wild water buffalo (Bubalus arnee) is an endangered species that is conserved in the Koshi Tappu Wildlife Reserve (KTWR), Nepal, and was recently translocated to the Chitwan National Park (CNP). Gastrointestinal (GI) parasites are the cause of significant negative health and production impacts on animals worldwide. Methods A coprological survey of GI parasites of wild water buffalo was carried out in the CNP in 2020. Fresh dung samples (n = 25) were collected from wild water buffaloes and analysed using sedimentation and flotation techniques for morphological identification of parasite cysts, oocysts and eggs. Results Nine different GI parasites were recorded of which Entamoeba spp. (20 samples, 80%) were the most common. The presence of Entamoeba spp. was further validated using polymerase chain reaction (PCR) analysis and DNA sequencing. The PCR results were positive for all of the microscopically positive samples, and the species was identified as Entamoeba bovis. Three samples were sequenced and formed a cluster of E. bovis, which was separated from other Entamoeba spp. in phylogenetic analysis. Conclusion This is the first report for molecular detection of E. bovis from wild water buffaloes in Nepal. Future work should focus on the prevalence of such infections in water buffaloes in forest environments. 1. The Asiatic wild water buffalo is an endangered species that are present only two areas in Nepal. 2. Microscopic examination revealed 96% prevalence rate with four protozoan and five helminths’ parasites detected in wild water buffalo. 3. PCR and sequencing revealed the presence of highly prevalent species as Entamoeba bovis. 4. This is the first molecular report of E. bovis in Nepalese wild water buffaloes.

Salmonella enterica subsp. enterica Serovar Enteritidis is one of the major pathogens associated with enteric diseases in animals and humans. Thus, due to the importance of Salmonella spp. infections for animal production and public health, the aim of the present study was to describe the first detection of S. enteritidis in an aborted water buffalo fetus in southern Italy by characterizing the phylogroup profile and the antimicrobial susceptibility of the isolated pathogenic strains. The different clinical manifestations of salmonellosis in animals include diarrhea, abortion, pneumonia, septic arthritis, meningitis, and others, depending on the virulence of the serovars, infectious dose, and host immunity. This study reports the first case of abortion caused by Salmonella enterica subsp enterica serovar Enteritidis in water buffalo ( Bubalus bubalis ) in the Campania region, southern Italy. Complete necropsy was performed on the aborted water buffalo fetus under study, and samples and swabs from different organs were collected. Samples were processed by microbiological and molecular analyses to detect bacterial, viral, and protozoarian pathogens possibly responsible for abortion. Whole genome sequencing (WGS) was carried out to further characterize the isolated S . Enteritidis strain. Our findings highlight the crucial role of S . Enteritidis as a potential abortive agent in water buffalo and its presence should therefore be investigated in cases of bubaline abortion.

Listeria monocytogenes (LM) is the causative agent of listeriosis in both animals and humans, representing one of the most severe food-borne diseases in humans. Out of 13 serotypes, only three (i.e., 1/2a, 1/2b, and 4b) are responsible for 95% of human outbreaks of listeriosis. Ruminants have been hypothesised to represent the main natural reservoir for this pathogen and to be involved in the transmission of Listeria to humans. During pregnancy, listeriosis in ruminants cause various reproductive disorders as well as abortion. However, little is known about abortion due to LM in water buffaloes ( Bubalus bubalis ). In this study, we report for the first time the detection of LM in a water buffalo foetus in the region of Campania, Italy. Complete necropsy was performed, and samples and swabs from the abomasum, kidneys, liver, lungs, and spleen were collected. Microbiological and molecular analyses were carried out to detect bacterial, viral, and protozoarian abortive pathogens. The results revealed the presence of LM in the liver, lungs, and abomasum, and no other agent was detected. Isolation was confirmed by biochemical and molecular tests. Molecular serotype characterisation was performed, and serogroup IVb was identified. In conclusion, because of the zoonotic implications of our findings, this report highlights the importance of including LM in the diagnostic panel in cases of bubaline abortion.

Four valid species of Sarcocystis have been reported from the water buffalo (Bubalus bubalis): Sarcocystis fusiformis, Sarcocystis buffalonis, Sarcocystis levinei and Sarcocystis dubeyi. Here, we redescribe structure of S. fusiformis sarcocysts by scanning and transmission electron microscopy (SEM, TEM). Twenty-one macroscopic sarcocysts from oesophagus of the water buffalo in Egypt were examined by light microscopy, SEM and TEM. The sarcocyst wall was up to 9 μm thick, depending on the section and the technique. In 5 μm paraffin-embedded sections, the sarcocyst wall was indistinct, 2–5 μm thick and appeared smooth. In 1 μm plastic-embedded sections stained with toluidine blue, the sarcocyst wall was 2·5–5·2 μm thick and had branched villar protrusions (vp)-like branches of a dead tree. By SEM, the sarcocyst wall had a mesh-like structure with irregularly shaped vp that were folded over the sarcocyst wall. On each vp there were uniform papillomatous structures that were 100 nm wide. By TEM, vp were up to 6 μm long and contained filamentous tubular structures, most of which were parallel to the long axis of the projections; granules were absent from these tubules. By TEM, bradyzoites within the same cyst varied from 11·2 to 16·8 μm in length. By TEM, bradyzoites had a very long (10 μm) convoluted mitochondrion, up to 12 dense granules, but only 2 rhoptries. This redescription should help to differentiate the sarcocysts of S. fusiformis from similar sarcocysts in domestic and wild ruminants.

Bovine tuberculosis (bTB) remains endemic in domestic water buffaloes ( Bubalus bubalis ) in India and elsewhere, with limited options for control other than testing and slaughter. The prescribed tuberculin skin tests with purified protein derivative (PPD) for diagnosis of bTB preclude the use of Bacille Calmette-Guérin (BCG)-based vaccination because of the antigenic cross-reactivity of vaccine strains with Mycobacterium bovis and related pathogenic members of the M. tuberculosis complex (MTBC). For the diagnosis of bTB in domestic water buffaloes, we here assessed a recently described defined-antigen skin test (DST) that comprises overlapping peptides representing the ESAT-6, CFP-10 and Rv3615c antigens, present in disease-causing members of the MTBC but missing in BCG strains. The performance characteristics of three doses (5, 10 or 20 μg/peptide) of the DST were assessed in natural tuberculin skin test reactor ( n = 11) and non-reactor ( n = 35) water buffaloes at an organized dairy farm in Hisar, India, and results were compared with the single intradermal skin test (SIT) using standard bovine tuberculin (PPD-B). The results showed a dose-dependent response of DST in natural reactor water buffaloes, although the SIT induced a significantly greater ( P < 0.001) skin test response than the highest dose of DST used. However, using a cut-off of 2 mm or greater, the 5, 10, and 20 μg DST cocktail correctly classified eight, 10 and all 11 of the SIT-positive reactors, respectively, suggesting that the 20 μg DST cocktail has a diagnostic sensitivity (Se) of 1.0 (95% CI: 0.72–1.0) identical to that of the SIT. Importantly, none of the tested DST doses induced any measurable skin induration responses in the 35 SIT-negative animals, suggesting a specificity point estimate of 1.0 (95% CI: 0.9–1.0), also identical to that of the SIT and compares favorably with that of the comparative cervical test (Se = 0.85; 95% CI: 0.55–0.98). Overall, the results suggest that similar to tuberculin, the DST enables sensitive and specific diagnosis of bTB in water buffaloes. Future field trials to explore the utility of DST as a defined antigen replacement for tuberculin in routine surveillance programs and to enable BCG vaccination of water buffaloes are warranted.