Potential utility of RSAD2 transcript and protein in early detection of pregnancy in buffaloes.

This study investigates a novel early pregnancy marker in water buffaloes, focusing on RSAD2 mRNA expression, known to be upregulated by interferon-tau (IFNT) during pregnancy. While RSAD2 is primarily recognized for its antiviral effect, we hypothesized its role as a conceptus-induced component in regulating pregnancy in buffaloes. Given its differential expression compared to other IFNT-induced genes in cows, RSAD2 may serve as a biomarker for early pregnancy detection in buffaloes. RNA, cDNA, and plasma samples were obtained from archived samples collected before insemination (d0) and at d20, d25 and d40 after insemination. Twelve RNA samples, having optimal optical density and concentration, from six pregnant and six non-pregnant buffaloes were selected. The cDNA was analyzed to measure the abundance of RSAD2 mRNA using real-time quantitative PCR (RT-qPCR) and plasma for protein expression analysis using Western blot. The RT-qPCR analysis showed a transcript of RSAD2 increased significantly by 7-fold and 6-fold on d20 and d25, compared to both d0 and d40 in the pregnant group only. At d20, the sensitivity of RSAD2 was 100% and the specificity was 83.3%, and at d25-d both the sensitivity and specificity was 100%, indicating low incidences of misdiagnosing early pregnancy in buffaloes. In the non-pregnant group, RSAD2 expression remained low and did not change after insemination. Western blot analysis revealed an immunoreactive RSAD2 protein band. Densitometry analysis of the RSAD2-specific protein band, based on gray mean value, showed significantly increased expression of RSAD2 at d25 compared to d0 in the pregnant group. In conclusion, these results indicated that RSAD2 expressions at both the mRNA and protein levels show promising potential for detecting pregnancy at d25 post-insemination.

Clinical Modalities for Enhancing Reproductive Efficiency in Buffaloes: A Review and Practical Aspects for Veterinary Practitioners.

This review aimed to bring a comprehensive analysis of key clinical strategies for enhancing reproductive efficiency in buffaloes, a species that exhibit low reproductive performance under conventional reproductive management compared to that exhibited by cattle. It considers key ART techniques including estrus synchronization for artificial insemination, and ovulation induction, highlighting their role in improving fertility and overall herd productivity. However, it also addresses common postpartum inflammatory and functional reproductive disorders, discussing their diagnosis and treatment protocols, stressing their impact on the overall reproductive outcome in buffalo farming.

Prevalence and Subtypes Distribution (ST10, ST14, ST25, ST26) of Blastocystis spp. in Anatolian Water Buffalo (Bubalus bubalis) in Van, Türkiye.

BACKGROUND: Blastocystis spp. is one of the most common protozoa worldwide, living in the gastrointestinal tract of humans and many other animals. On the basis of the genetic heterogeneity of small subunit ribosomal RNA, at least 28 subtypes (ST1-ST17, ST21 and ST23-ST32) are reported to exist in mammals and birds. OBJECTIVES: This study was carried out to determine the prevalence and subtypes of Blastocystis spp. in Anatolian buffaloes (Bubalus bubalis) in Van province in the Eastern Anatolia Region of Turkey. METHODS: DNA was extracted using commercial GeneMATRIX Stool DNA Purification Kit and then stored at -20°C until PCR amplification. After PCR amplification of the SSU rRNA gene region positive Blastocystis spp., amplicons from buffalo faeces were sequenced and then deposited in GenBank (OR576949.1, OR576950.1, OR576970.1, OR576971.1, OR577019.1, PP837943.1, PP837940.1, PP837939.1, PP837604.1, PP837937.1, PP837934.1, PP837601.1, PP837936.1 and PP837603.1). RESULTS: PCR analysis of 120 faecal samples showed a total prevalence of 11.67% (14/120). The prevalence was higher in females and young animals (p > 0.05). Sequence analysis revealed Blastocystis spp., ST10, ST14, ST25 and ST26 subtypes. To our knowledge, Blastocystis subtypes ST25 and ST26 in buffaloes were reported for the first time in this study. CONCLUSION: It is thought that more large-scale studies should be carried out to determine the zoonotic subtype potential of this protozoan in the region.

Prevalence of Neospora caninum and associated risk factors in semi-intensive female water buffaloes reared in Greece, using an ELISA kit on milk samples.

Neospora caninum is an apicomplexan (family: Sarcocystidae) protozoan parasite with a global distribution. In the N. caninum life cycle, dogs and other related canids are the definitive hosts, while other animals such as water buffaloes (Bubalus bubalis) constitute the intermediate host for this parasite. In many countries, the water buffalo is of high economic importance, providing valuable high-quality products for human needs. Although knowledge concerning the prevalence of this parasite in intermediate animal host populations is crucial, data from water buffalo are scarce. Keeping this in mind, the aim of this study was to examine the presence and assess the prevalence rates of N. caninum infection in water buffaloes in Northern Greece, where this animal husbandry sector started to raise, as well as to determine associated risk factors, with the application of a commercially available Neospora ISCOM ELISA test kit, developed for the detection of antibodies against N. caninum in milk samples The study was conducted during January-June 2023 in a total of 172 individual raw milk samples, collected from dairy buffaloes, reared under a semi extensive system, in three different farms. Information on the status of N. caninum infection in buffaloes from Greece was so far unknown with a lack of epidemiological or risk factors associated. For the detection of N. caninum, the commercially available Neospora ISCOM ELISA test kit (SANOVIR ®, Svanova Biotech AB, Uppsala, Sweden) was utilized. Results demonstrated the presence of N. caninum in water buffaloes from Greece for the first time. All positive N. caninum animal were asymptomatic, with absence of any disease signs. The overall prevalence of infection was 22.10%, whereas the main risk factors include the presence of dogs as well as the low biosecurity measures.

Disentangling river and swamp buffalo genetic diversity: initial insights from the 1000 Buffalo Genomes Project.

More people in the world depend on water buffalo for their livelihoods than on any other domesticated animals, but its genetics is still not extensively explored. The 1000 Buffalo Genomes Project (1000BGP) provides genetic resources for global buffalo population study and tools to breed more sustainable and productive buffaloes. Here we report the most contiguous swamp buffalo genome assembly (PCC_UOA_SB_1v2) with substantial resolution of telomeric and centromeric repeats, ∼4-fold more contiguous than the existing reference river buffalo assembly and exceeding a recently published male swamp buffalo genome. This assembly was used along with the current reference to align 140 water buffalo short-read sequences and produce a public genetic resource with an average of ∼41 million single nucleotide polymorphisms per swamp and river buffalo genome. Comparison of the swamp and river buffalo sequences showed ∼1.5% genetic differences, and estimated divergence time occurred 3.1 million years ago (95% CI, 2.6-4.9). The open science model employed in the 1000BGP provides a key genomic resource and tools for a species with global economic relevance.

Analysis of miRNAs in milk of four livestock species.

BACKGROUND: Milk is essential for mammalian nutrition because it provides vital nutrients for growth and development. Milk composition, which is influenced by genetic and environmental factors, supports lactation, a complex process crucial for milk production and quality. Recent research has focused on noncoding RNAs, particularly microRNAs (miRNAs), which are present in body fluids and regulate gene expression post-transcriptionally. This study comprehensively characterizes miRNAs in milk of four livestock species, namely Bubalus bubalis, Capra hircus, Equus asinus, and Ovis aries and identifies potential target genes. RESULTS: High-throughput sequencing of milk RNA resulted in distinct read counts across species: B. bubalis (8,790,441 reads), C. hircus (12,976,275 reads), E. asinus (9,385,067 reads), and O. aries (7,295,297 reads). E. asinus had the highest RNA mapping rate (94.6%) and O. aries the lowest (84.8%). A substantially greater proportion of miRNAs over other small RNAs was observed for the donkey milk sample (7.74%) compared to buffalo (0.87%), goat (1.57%), and sheep (1.12%). Shared miRNAs, which included miR-200a, miR-200b, miR-200c, and miR-23a among others, showed varying expression levels across species, confirmed by qPCR analysis. Functional annotation of predicted miRNA target genes highlighted diverse roles, with an enrichment in functions linked to metabolism and immunity. Pathway analysis identified immune response pathways as significant, with several miRNAs targeting specific genes across species, suggesting their regulatory function in milk. CONCLUSIONS: Both conserved and species-specific miRNAs were detected in milk of the investigated species. The identified target genes of these miRNAs have important roles in neonatal development, adaptation, growth, and immune response. Furthermore, they influence milk and meat production traits in livestock.

Effect of histidine and L-Tyrosine supplementation in maturation medium on in-vitro developmental outcomes of buffalo oocytes.

The present study was designed to investigate the effects of amino acid (histidine and L-Tyrosine) on in vitro maturation (IVM), in vitro fertilization (IVF), cleavage (CR) rates, and in vitro embryonic cultivation (IVC; Morula and Blastocyst stage) in buffaloes. Within two hours of buffalo slaughter, the ovaries were collected and transported to the laboratory. Follicles with a diameter of 2 to 8 mm were aspirated to recover the cumulus oocyte complexes (COCs). Histidine (0.5, 1, and 3 mg/ml) or L-Tyrosine (1, 5, and 10 mg/ml) were added to the synthetic oviductal fluid (SOF) and Ferticult media. The IVM, IVF, CR, and IVC (Morula and Blastocyst) rates were evaluated. The results showed that SOF maturation media containing histidine at 0.5 mg/ml significantly (P ≤ 0.01) improved the oocyte maturation when compared to control and other concentrations. The addition of histidine to FertiCult media at 0.5, 1, and 3 mg/ml did not improve the IVM, IVF, CR, or IVC percentages. However, the embryos in the control group were unable to grow into a morula or blastocyst in the SOF or Ferticult, while addition of L-Tyrosine to the SOF or Ferticult at various concentrations improved IVC (morula and blastocyst rates). There was a significant (P ≤ 0.01) increase in IVM when histidine was added to SOF medium at a concentration of 0.5 mg/ml compared with L-Tyrosine. Also, there were significant (P ≤ 0.01) increases in IVC when L-Tyrosine was added to SOF medium at concentrations of 1 and 10 mg/ml compared with histidine. In conclusion, the supplementation of the SOF and FertiCult with the amino acids histidine and L-Tyrosine improve the maturation rate of oocytes and development of in vitro-produced buffalo embryos.

Bison mother-offspring acoustic communication.

Mother-offspring communication is especially crucial for social species in order to synchronize activities essential for early survival including nursing, resting, maintaining proximity during group movements between food or water sources, and locating one another if separated in a large social group. One of the most social ungulate species in North America is the American Bison (Bison bison), formerly known as buffalo. Adult female bison associate with their young for over a year and communication between mother and offspring is likely essential for establishing and maintaining a bond upon which the life of a calf depends. One goal of this study was to quantify and compare the acoustic form of vocalizations of adult female, subadult, and calf bison and to determine how age classes differed in call structure. The other goal was to identify the contexts in which bison vocalized. Vocalizations of 101 bison (53 adult females, 15 subadults, 33 calves) in a semi-free-ranging herd in Montana were analyzed and found to be pulsatile sounds, unlike vocalizations of bison bulls or domestic cows and calves. Vocalizations of bison cows, subadults, and calves differed significantly in total duration, numbers of pulses, pulse duration, and pulse rate. Seven distinct call contexts were identified. The majority of calls were "moving-on calls" (39%), when a cow called and her calf ran to her side and the 2 moved on together, and "contact calls" (21%) when a cow called and her calf called back but neither changed their location. "Imprinting calls" and "nursing calls" were also identified. Mother-offspring acoustic communication in bison appears especially critical for coordinating movements. Understanding the role of acoustic communication in maintaining the bond between bison mothers and their offspring can contribute to the humane management and welfare of this iconic species.

La comunicación madre-cría es especialmente crucial para especies sociales, debido a que permite sincronizar actividades esenciales para la supervivencia temprana, como la lactancia, el descanso, el mantenimiento de proximidad durante los movimientos de los grupo entre las fuentes de alimento o agua y la localización mutua en caso de separación dentro de un grupo social grande. Una de las especies de ungulados más sociales de América del Norte es el bisonte (Bison bison). Las hembras adultas de bisonte se asocian con sus crías durante más de un año y la comunicación entre madre el becerro es probablemente esencial para establecer y mantener un vínculo en el que depende la vida de la cría. Uno de los objetivos de este estudio fue cuantificar y comparar la forma acústica de las vocalizaciones de hembras adultas, subadultas y crías de bisonte, y determinar cómo diferían las clases por edad en la estructura de las llamadas. Otro objetivo fue identificar los contextos en los que se emitían las vocalizaciones. Se analizaron las vocalizaciones de 101 bisontes (53 hembras adultas, 15 hembras subadultas, 33 crías) en un rebaño semi-libre en Montana. Se encontró que estas vocalizaciones eran sonidos pulsátiles, completamente diferentes a los emitidos por los bisontes machos adultos o las vacas y becerros domésticos. Las vocalizaciones diferían significativamente entre las tres clases de edad en su duración total, número de pulsos, duración de los pulsos y ritmo de los pulsos. La mayoría de las llamadas se dieron en dos contextos: "llamadas de avance" (39%), cuando una hembra adulta llamaba y su cría corría a su lado y ambas avanzaban juntas, y "llamadas de contacto" (21%), cuando una hembra adulta llamaba y su cría respondía, pero ninguna cambiaba su ubicación. También se identificaron "llamadas de impronta" y "llamadas de amamantamiento," así como otros tres contextos de llamada. La comunicación acústica madre-cría en bisontes parece especialmente crítica para coordinar los movimientos. Entender el papel de la comunicación acústica en el mantenimiento del vínculo entre las madres y sus crías puede contribuir al manejo humanitario y al bienestar de esta especie icónica. Este trabajo representa el primer estudio que investiga cuantitativamente las señales acústicas de hembras adultas, subadultas y crías de bisontes Norte Americanos mientras se desplazan en condiciones de semi-libertad.

In Silico Analysis: Molecular Characterization and Evolutionary Study of CLCN Gene Family in Buffalo.

Chloride channels (ClCs) have received global interest due to their significant role in the regulation of ion homeostasis, fluid transport, and electrical excitability of tissues and organs in different mammals and contributing to various functions, such as neuronal signaling, muscle contraction, and regulating the electrolytes' balance in kidneys and other organs. In order to define the chloride voltage-gated channel (CLCN) gene family in buffalo, this study used in silico analyses to examine physicochemical properties, evolutionary patterns, and genome-wide identification. We identified eight CLCN genes in buffalo. The ProtParam tool analysis identified a number of important physicochemical properties of these proteins, including hydrophilicity, thermostability, in vitro instability, and basic nature. Based on their evolutionary relationships, a phylogenetic analysis divided the eight discovered genes into three subfamilies. Furthermore, a gene structure analysis, motif patterns, and conserved domains using TBtool demonstrated the significant conservation of this gene family among selected species over the course of evolution. A comparative amino acid analysis using ClustalW revealed similarities and differences between buffalo and cattle CLCN proteins. Three duplicated gene pairs were identified, all of which were segmental duplications except for CLCN4-CLCN5, which was a tandem duplication in buffalo. For each gene pair, the Ka/Ks test ratio findings showed that none of the ratios was more than one, indicating that these proteins were likely subject to positive selection. A synteny analysis confirmed a conserved pattern of genomic blocks between buffalo and cattle. Transcriptional control in cells relies on the binding of transcription factors to specific sites in the genome. The number of transcription factor binding sites (TFBSs) was higher in cattle compared to buffalo. Five main recombination breakpoints were identified at various places in the recombination analysis. The outcomes of our study provide new knowledge about the CLCN gene family in buffalo and open the door for further research on candidate genes in vertebrates through genome-wide studies.

Preovulatory follicular dynamics and ovulatory events following the use of GnRH 84 h after medroxyprogesterone acetate sponge removal in postpartum buffaloes.

Herein, we evaluated the effects of Gonadotropin hormone-releasing hormone (GnRH) administration 84 h after medroxyprogesterone acetate (MAP) sponge removal on follicular growth, ovulation timing, and pregnancy per artificial insemination (AI) in cosynchronized postpartum Nili Ravi buffaloes. In this study, 58 Nili Ravi postpartum buffaloes (DIM = 103 ± 1.64) were randomly divided into two treatment groups (n = 29/treatment): GnRH-TAI-84 and TAI-84. All buffaloes were administered a MAP sponge for seven days. Upon MAP sponge removal, all the subjects received prostaglandin F2α (PGF2α), and Timed AI (TAI) was performed 84 hours after sponge removal. In the GnRH-TAI-84 group, the buffaloes received GnRH alongside insemination, whereas in the TAI-84 group, the buffaloes were inseminated without GnRH administration. Follicle diameter and blood estradiol levels were measured every 6 h from 72-108 h after MAP sponge removal. The animals were checked for pregnancy using ultrasonography 40 days after AI. Animals subjected to the GnRH-TAI-84 protocol had a higher follicular growth rate and preovulatory follicle size than those in the TAI-84 group. The follicular diameter was also larger in animals that received GnRH-TAI-84 than in those that received TAI-84 90 and 96 h after MAP sponge removal. Buffaloes in the GnRH-TAI-84 group had lower estradiol concentrations at 90, 96, 102, and 108 h than those in the TAI-84 group. Ovulation in GnRH-TAI-84 buffaloes occurred 11 h earlier than that in buffaloes from the TAI-84 group. A shorter interval between AI and ovulation in GnRH-TAI-84 buffaloes (14 h vs. 25 h) led to greater pregnancies per AI (62% vs. 17%) compared to buffaloes from the TAI-84 group.

Quantitative proteomics reveals significant variation in host responses of cattle with differing buffalo fly susceptibility.

Background: Control of buffalo flies (Haematobia irritans exigua, BFs) relies mainly on chemical methods; however, resistance to insecticides is widespread in BF populations. Breeding for resistance to BFs represents a possible alternative, but direct phenotyping of animals is laborious and often inaccurate. The availability of reliable diagnostic biomarker(s) to identify low BF carrier cattle would facilitate rapid and accurate selection for genetic improvement. However, limited information is available regarding differences amongst cattle in host responses to BF infestation. Methods: This study investigated the variation in Brangus cattle serum proteomic profiles before (naïve) and after peak BF exposure, in low (LF) and high BF burden (HF) cattle. Cattle were phenotyped for susceptibility based on BF counts on multiple dates using visual and photographic techniques. The relative abundance of serum proteins in cattle before and after exposure to BFs was analysed using sequential window acquisition of all theoretical fragment ion mass spectrometry (SWATH-MS). Results: Exposure to BFs elicited similar responses in HF and LF cattle, with 79 and 70 proteins, respectively, showing significantly different abundances post exposure as compared to their relevant naïve groups. The comparison of serum samples from naïve HF and LF cattle identified 44 significantly differentially abundant (DA) proteins, while 37 significantly DA proteins were identified from the comparison between HF and LF cattle post-exposure to BFs. Proteins with higher abundance in naïve LF cattle were enriched in blood coagulation mechanisms that were sustained after exposure to BFs. Strong immune response mechanisms were also identified in naïve LF cattle, whereas these responses developed in HF cattle only after exposure to BF. High BF cattle also showed active anticoagulation mechanisms in response to BF exposure, including downregulation of coagulation factor IX and upregulation of antithrombin-III, which might facilitate BF feeding. Conclusion: Underlying differences in the abundance of proteins related to blood coagulation and immune response pathways could potentially provide indirect indicators of susceptibility to BF infestation and biomarkers for selecting more BF-resistant cattle.

Genetic and Phenotypic Trends for Production and Reproduction Traits in Murrah Buffaloes.

The objective of the study was to estimate genetic and phenotypic trends for first lactation production and reproduction traits in Murrah buffaloes. The information of pedigree and targeted traits of 640 Murrah buffaloes was collected for the period from 1997 to 2020. The first lactation production traits included first lactation milk yield (FLMY), 305 days first lactation milk yield (305FLMY), first lactation length (FLL), first lactation peak yield (FPY) whereas reproduction traits included first service period (FSP), first calving interval (FCI) and first dry period (FDP). Genetic and phenotypic trends were estimated. Phenotypic trends for FLMY, 305FLMY, FLL and FPY exhibited as 36.96 ± 8.58 kg/year, 31.93 ± 8.34 kg/year, 1.47 ± 0.55 days/year and 0.12 ± 0.02 kg/year, respectively and respective genetic trends exhibited as 3.73 ± 1.67 kg/year, 1.94 ± 0.76 kg/year, -0.15 ± 0.07 days/year and 0.01 ± 0.01 kg/year, respectively. It was revealed that there were significant (p < 0.05) and positive phenotypic trends for all production traits while genetic trends were significant (p < 0.05) for FLMY and 305FLMY traits. The phenotypic trends of studied reproductive traits indicated that only FDP trait significantly (p < 0.01) decreased (1.87 days/year) over time. For FSP and FCI traits, nonsignificant (p > 0.05) genetic and phenotypic trends indicated no change over time. This study highlighted potential challenges in maintaining reproductive efficiency alongside productivity improvements in Murrah buffaloes.

Meta-Genomic Analysis of Different Bacteria and Their Genomes Found in Raw Buffalo Milk Obtained in Various Farms Using Different Milking Methods.

Milking methods have significant impacts on the microbiological composition, which could affect the quality of raw buffalo milk. Hence, the current study was conducted on the impact of milking methods on microorganisms in buffalo tank raw milk from 15 farms in Guangxi, China. The farms were divided into two groups based on the milking method: mechanical milking (MM, n = 6) and hand milking (HM, n = 9). Somatic cell counts, bacterial cell counts and nutrients of the raw buffalo milk samples were analyzed. The comparison of raw buffalo milk samples was analyzed using metagenomic sequencing to detect any differences between the two groups. There was no significant difference in the basic nutritional compositions and somatic cell count of raw buffalo milk between the two milking methods. However, the HM samples had significantly higher bacterial counts and diversity compared to the MM samples. The results showed that Staphylococcus spp., Klebsiella spp., Streptococcus spp., and Pseudomonas spp. were the major microbes present in canned raw buffalo milk. However, the differences between the two milking methods were the relative abundance of core microorganisms and their potential mastitis-causing genera, including the content of antibiotic-resistance genes and virulence genes. Our study revealed that Staphylococcus spp. and Streptococcus spp. were significantly more abundant in the MM group, while Klebsiella spp. was more abundant in the HM group. Regardless of the milking method used, Pseudomonas spp. was identified as the primary genus contributing to antibiotic resistance and virulence genes in canned raw buffalo milk. These findings affirm that there are differences in the microbial and genomic levels in canned raw milk. To prove the functional roles of the discovered genes and how these genes affect milk quality, further research and experimental validation are necessary.

Follicular Atresia in Buffalo: Cocaine- and Amphetamine-Regulated Transcript (CART) and the Underlying Mechanisms.

Atresia is a process in ovarian follicles that is regulated by hormone-induced apoptosis. During atresia, granulosa cell (GC) apoptosis is a key mechanism orchestrated through diverse signaling pathways. Cocaine- and amphetamine-regulated transcript (CART) signaling within ovarian GCs has been demonstrated to play a key role in the regulation of follicular atresia in cattle, pigs, and sheep. The present work aimed to investigate the potential local regulatory role of CART in GC apoptosis-induced follicular atresia in buffalo, focusing on the modulation of the AKT/GSK3ß/ß-catenin signaling pathways, which are the intracellular signaling pathways involved in cell viability. Our findings revealed increased expression of CARTPT and BAX and decreased levels of AKT, ß-catenin, and CYP19A1 genes in atretic follicles compared to healthy follicles. Subsequently, CART treatment in the presence of FSH inhibited the FSH-induced increase in GC viability by reducing estradiol production and increasing apoptosis. This change was accompanied by an increase in the gene expression levels of both CARTPT and BAX. At the protein level, treatment with CART in the presence of FSH negatively affected the activity of AKT, ß-catenin, and LEF1, while the activity of GSK3ß was enhanced. In conclusion, our study shows how CART negatively influences buffalo GC viability, underlying the modulation of the AKT/GSK3ß/ß-catenin pathway and promoting apoptosis-a key factor in follicular atresia.

Differences in Milk Fatty Acids Profile of Two Breeds of Water Buffaloes Explained by Their Gastrointestinal Microbiota.

This experiment investigated gastrointestinal microbes' role in milk fatty acid differences between Murrah and Nili-Ravi buffaloes. After 30 days of a basal diet, rumen microbial diversity was similar, but Murrah buffaloes had greater partially unsaturated fatty acids like C18:2c9t11. Rumen bacteria like Acetobacter, Ruminococcus, and Prevotellaceae_YAB2003_group correlated positively with milk fatty acids C22:5n-6 and C18:3 in Murrah. Fecal microbial beta diversity differed, with UCG-005 and Prevolla positively correlated with C18:2c9t11 and C22:5n-6. The greater quantity of milk fatty acids C18:3, C18:2c9t11, and C22:5n-6 in Murrah milk was linked to rumen and fecal microbes. This suggests that gastrointestinal microbes like Acetobacter, Ruminococcus, and UCG_005 regulate milk fatty acid concentrations in buffaloes.

Diagnostic Accuracy of Non-Contrast-Enhanced Time-Resolved MR Angiography to Assess Angioarchitectural Classification Features of Brain Arteriovenous Malformations.

This study aims to assess the diagnostic accuracy of non-contrast-enhanced 4D MR angiography (NCE-4D-MRA) compared to contrast-enhanced 4D MR angiography (CE-4D-MRA) for the detection and angioarchitectural characterisation of brain arteriovenous malformations (bAVMs). Utilising a retrospective design, we examined 54 MRA pairs from 43 patients with bAVMs, using digital subtraction angiography (DSA) as the reference standard. Both NCE-4D-MRA and CE-4D-MRA were performed using a 3-T MR imaging system. The primary objectives were to evaluate the diagnostic performance of NCE-4D-MRA against CE-4D-MRA and DSA and to assess concordance between imaging modalities in grading bAVMs according to four main scales: Spetzler-Martin, Buffalo, AVM embocure score (AVMES), and R2eDAVM. Our results demonstrated that NCE-4D-MRA had a higher accuracy and specificity compared to CE-4D-MRA (0.85 vs. 0.83 and 95% vs. 85%, respectively) and similar agreement, with DSA detecting shunts in bAVMs or residuals. Concordance in grading bAVMs was substantial between NCE-4D-MRA and DSA, particularly for the Spetzler-Martin and Buffalo scales, with CE-4D-MRA showing slightly higher kappa values for interobserver agreement. The study highlights the potential of NCE-4D-MRA as a diagnostic tool for bAVMs, offering comparable accuracy to CE-4D-MRA while avoiding the risks associated with gadolinium-based contrast agents. The safety profile of imaging techniques is a significant concern in the long-term follow up of bAVMs, and further prospective research should focus on NCE-4D-MRA protocol improvement for clinical use.

Effect of season on histoarchitecture of pineal gland in buffalo (Bubalus bubalis).

Background: The photoperiod and other seasonal variations are the key factors that affect reproduction and production of the animals. The pineal gland secretes melatonin hormone that affects several physiological functions of the body during different seasons. Aims: The present study was conducted to study the histoarchitectural and micrometrical changes in the pineal gland of buffalo (Bubalus bubalis) during different seasons of the year. Methods: Pineal glands of 30 adult female Jaffarabadi buffaloes were collected from the slaughterhouse during the winter, summer, and rainy seasons. Samples were processed by standard histological procedures and stained with various stains for histological and micrometrical observations. Results: The pinealocytes constituted a major cellular portion of pineal parenchyma. The pinealocyte nuclei were lightly stained and more euchromatic during the winter season whereas darkly stained and slightly heterochromatic during summer. The calcium deposits occupied a larger area of pineal parenchyma during the summer as compared to the winter season. The pinealocyte density, the nuclear diameter of pinealocytes, and the number of argyrophilic nucleolar organizer regions (AgNOR) were highest during the winter season as compared to the summer and rainy seasons. Conclusion: The present study shows the influence of season on the histoarchitecture and histometry of the pineal gland of buffalo and indicated higher pineal activity during the winter season in this species.

The influence of rutin and chlorogenic acid on oxidative stress and in vivo fertility: Evaluation of the quality and antioxidant status of post-thaw semen from Azari water buffalo bulls.

BACKGROUND: The vulnerability of buffalo sperm to cryoinjury necessitates the improvement of sperm cryo-resistance as a critical strategy for the widespread use of assisted reproductive technologies in buffalo. OBJECTIVES: The main aim of the present study was to evaluate the effects of different concentrations of rutin and chlorogenic acid (CGA) on buffalo semen quality, antioxidant activity and fertility during cryopreservation. METHODS: The semen was collected and pooled from the 3 buffaloes using an artificial vagina (18 ejaculations). The pooled sperm were divided into nine different groups: control (Tris-based extender); 0.4, 0.6, 0.8 and 1 mM rutin (rutin + Tris-based extender); and 50, 100, 150 and 200 µM CGG (CGA + Tris-based extender). Sperm kinematics, viability, hypo-osmotic swelling test, mitochondrial activity, antioxidant activities and malondialdehyde (MDA) concentration of frozen and thawed buffalo sperm were evaluated. In addition, 48 buffalo were finally inseminated, and pregnancy was rectally determined 1 month after insemination. RESULTS: Compared to the control group, adding R-0.4, R-0.6, CGA-100 and CGA-150 can improve total and progressive motility, motility characteristics, viability, PMF and DNA damage in buffalo sperm. In addition, the results showed that R-0.4, R-0.6, CGA-50, CGA-100 and CGA-150 increased total antioxidant capacity, catalase, glutathione peroxidase and glutathione activities and decreased MDA levels compared to the control group. Furthermore, it has been shown that adding 150 µM CGA and 0.6 mM rutin to an extender can increase in vivo fertility compared to the control group. CONCLUSIONS: In conclusion, adding rutin and CGA to the extender improves membrane stability and in vivo fertility of buffalo sperm by reducing oxidative stress.

Estimation of the threshold for heat stress and genetic features for milk yield in Mehsana buffaloes in India.

Heat stress is one of the primary environmental factors that harm both the productivity and health of buffaloes. The current study was conducted to estimate the threshold of temperature humidity index (THI)1 and genetic features for milk yield of first-lactation Mehsana buffaloes using an univariate repeatability test-day model. The data included 130,475 first lactation test-day milk yield (FLTDMY) records of 13,887 Mehsana buffaloes and the daily temperature and humidity. The statistical model included herd test day as fixed effects, days-in-milk (DIM) classes, age of the animal, as well as random factors such as the additive genetic effect (AGE) of animal in general conditions (intercept), AGE of the buffaloes subjected to heat stress (slope), permanent environmental effect of animal in general conditions (intercept), permanent environmental effect of animal under heat stress conditions (slope) and random residual effect. It was expected that the general effects and the heat-tolerance effects would be correlated, represented by the present investigation's repeatability models. The variance components of FLTDMY in the present study were computed using the REML method. The threshold for THI was 78. At the THI below the threshold, the heritability estimated for the FLTDMY trait was 0.29, and the additive genetic variance (AGV) for heat stress conditions was 0. At THI of 83, AGV for heat stress conditions was highest for FLTDMY. The genetic correlation of general AGE to heat-tolerant AGE was -0.40. The results indicated that a consistent selection for milk production, avoiding the thermal tolerance, may diminish the thermal tolerance capacity of Mehsana buffaloes.

CX43 and oxidative stress are the targets of BCB staining to predict the developmental potential of buffalo oocytes.

This study used the brilliant cresyl blue (BCB) staining method to group buffalo oocytes (BCB+ and BCB-) and perform in vitro maturation, in vitro fertilization and embryo culture. At the same time, molecular biology techniques were used to detect gap junction protein expression and oxidative stress-related indicators to explore the molecular mechanism of BCB staining to predict oocyte developmental potential. The techniques of buffalo oocytes to analyse their developmental potential and used immunofluorescence staining to detect the expression level of CX43 protein, DCFH-DA probe staining to detect ROS levels and qPCR to detect the expression levels of the antioxidant-related genes SOD2 and GPX1. Our results showed that the in vitro maturation rate, embryo cleavage rate and blastocyst rate of buffalo oocytes in the BCB+ group were significantly higher than those in the BCB- group and the control group (p < .05). The expression level of CX43 protein in the BCB+ group was higher than that in the BCB- group both before and after maturation (p < .05). The intensity of ROS in the BCB+ group was significantly lower than that in the BCB- group (p < .05), and the expression levels of the antioxidant-related genes SOD2 and GPX1 in the BCB+ group were significantly higher than those in the BCB- group (p < .05). Brilliant cresyl blue staining could effectively predict the developmental potential of buffalo oocytes. The results of BCB staining were positively correlated with the expression of gap junction protein and antioxidant-related genes and negatively correlated with the reactive oxygen species level, suggesting that the mechanism of BCB staining in predicting the developmental potential of buffalo oocytes might be closely related to antioxidant activity.