Search Results (22)

Nili-Ravi buffalo is a core dairy breed for China’s genetic improvement, yet its assisted reproductive technologies are restricted by low ovum pick-up (OPU) efficiency and unstable embryo transfer (ET) pregnant rates. This study established statistically optimized key parameters for OPU procedures in Nili-Ravi buffalo, and evaluated the effects of multiple ET-related protocols. For OPU, 50–55 mmHg vacuum pressure balanced oocyte recovery and usable rate best (p < 0.05). Young and adult buffalo performed significantly better than prepubertal heifers (p < 0.05). Four consecutive weekly consecutive OPU sessions did not significantly reduce oocyte yield (p > 0.05). FSH treatment markedly enhanced oocyte quantity, quality and recovery efficiency (p < 0.05), with no significant difference between two FSH protocols (p > 0.05). For ET, three estrus synchronization protocols showed no significant effects on estrus, ovulation, transferable recipient and pregnant rates in breeding or non-breeding seasons (p > 0.05), though optimized protocols showed a numerical improvement. Combinations of CL grade and embryo quality, as well as post-transfer treatments with vitamin ADE, hCG and progesterone, did not significantly affect pregnancy or early abortion rates (p > 0.05). This study built a practical reference framework for OPU-ET procedures in Nili-Ravi buffalo, with key OPU parameters statistically optimized and ET protocols showing numerical improvements that require further validation with larger sample sizes. Full article

The river buffalo is an economically important livestock species supplying milk and meat. However, a multi-tissue transcriptomic atlas for the key dairy river buffalo breeds, Murrah and Nili-Ravi, has not yet been established, and the lack of stable reference genes has hindered in-depth studies of their biological functions and the molecular mechanisms underlying key economic traits such as lactation. We established a multi-tissue gene expression atlas across 20 tissues and identified 717 housekeeping genes (HKGs), and RPL37A and EEF2 were further shown to be stable candidate reference genes under the conditions tested. We found 8368 tissue-specific genes (TSGs), predominantly enriched in the reproductive system. Exploratory analysis of mammary tissue (dry-period vs. public lactating samples, confounded by batch effects) revealed mammary-enriched hub genes including LALBA; these findings are preliminary and require validation. Dynamic analysis across lactation stages (early, peak, mid-, late) identified candidate genes including SEC14L2 and ACSM3. Phenotypic data showed strong negative correlations between milk yield and protein/fat content, and a positive correlation with lactose content. However, causal or regulatory roles were not inferred due to lack of paired individual-level data. Cross-dataset comparisons are descriptive only, and are not key conclusions. In summary, this study lays the foundation for advancing research in lactation trait genetics and functional genomics in river buffalo, with novel reference genes and lactation stage-specific transcriptional dynamics as its main contributions. Full article

Melatonin (MLT) is a potent antioxidant that reduces oxidative stress (OS)-induced sperm damage. However, few studies have explored its effects in the field investigated here. This study aimed to evaluate the effects of MLT supplementation in extenders used for staining, sorting, and freezing on the quality of sorted Nili-Ravi buffalo sperm, embryo development after in vitro fertilization (IVF), and pregnancy outcomes following artificial insemination (AI). Computer-aided sperm analyzer (CASA) showed that progressive motility (PM) and velocity parameters of sorted, frozen–thawed sexed sperm were higher in the MLT-treated groups (p < 0.05). Fluorescence microscopy demonstrated that MLT significantly improved sperm plasma membrane integrity after staining and increased the proportion of frozen–thawed sex-sorted sperm with high mitochondrial membrane potential (HMMP) (p < 0.05). Raman spectroscopy further identified several distinctive Raman bands at 936, 1300, and 1651–1652 cm⁻¹, which may serve as biomarkers for assessing sperm quality. Additionally, cleavage rate, blastocyst formation, and pregnancy rates following IVF and AI were higher in the MLT-treated group (p < 0.05). In conclusion, MLT can serve as a valuable additive during sperm sex-sorting procedures to enhance sperm quality, thereby improving embryo developmental competence and pregnancy outcomes. Full article

Buffaloes (Bubalus bubalis) are central to the dairy and agricultural economy, contributing high-quality milk, meat, draft power, and manure. Rich milk composition, the ability to utilize low-quality roughage, and strong disease resistance make buffaloes indispensable across diverse production systems. Among India’s major dairy breeds—Murrah, Nili-Ravi, Jaffarabadi, Surti, Bhadawari, Mehsana, and Nagpuri, none exhibit the distinctive trait of intermittent milking, which is uniquely observed in the Pandharpuri buffalo, a registered indigenous breed of Maharashtra. Despite coexisting with dominant dairy breeds such as Murrah, Pandharpuri buffalo is considered to possess primitive riverine ancestry and may represent one of the ancestral lineages from which several Indian breeds evolved. Its evolutionary relevance and unique intermittent milking capacity underscore the need to understand its genomic architecture. To address this, we applied whole-genome resequencing and the De-Correlated Composite of Multiple Signals (DCMS) approach to identify within-breed selection signatures. Our analyses identified 1337 candidate genes, including several linked to milk production, particularly those relevant to the physiological capacity for intermittent milking. Notable genes included ERBB4, ESR1, SYK, INSR, PTPN11, VAV3, MAPK3, and PRKG1. These signatures provide insights into genomic regions and biological pathways that may be involved in lactation-related processes relevant to intermittent milking. The identified genomic regions offer promising targets for functional validation and future genome-informed breeding strategies aimed at conserving this unique indigenous germplasm while improving lactation efficiency and resilience. Full article

This study investigated the association between temperament and retrospective growth rates in 84 Nili Ravi buffalo heifers aged 18 to 24 months. Temperament was assessed using chute score and exit velocity, measured twice at a seven-day interval, and classified as calm (≤3) or nervous (>3). Retrospective average daily weight gain data were retrieved from farm records, and blood samples were collected to measure cortisol levels. ANOVA was used to analyze data, considering temperament, age group, season, and year of birth as fixed effects, with birth weight as a covariate. Results showed that 48 heifers were calm and 36 were nervous. Calm heifers exhibited significantly higher average daily gains than nervous heifers during the post-weaning period, with an increase of 240 g/day from 4 to 6 months and 190 g/day from 6 to 12 months (p < 0.001). However, this difference was not significant at 18–24 months (p = 0.144). Calm heifers have numerically lower cortisol levels (0.96 vs. 1.27 μg/dL; p = 0.11). These findings suggest that calmer heifers grow faster in early life, emphasizing the importance of temperament in breeding programs aimed at improving growth performance and welfare. Full article

This study aimed to evaluate the crude protein (CP) degradation kinetics and degradability (CPD) of cereal and legume fodder species grown at two geographically distant locations. Ten forage species, comprising six cereals (barley, maize, millet, oats, sorghum, and wheat) and four legumes (berseem, jantar, lucerne, and mustard), were evaluated to determine the effects of forage family, species, and location of growth on CP degradation fractions and effective CPD. The forage crops were cultivated under uniform agronomic practices at two distinct agro-ecological locations and were harvested at the booting stage (cereals) and 50% flowering stage (legumes). Dried and ground samples were incubated in the rumen of four Nili-Ravi buffalo fitted with rumen cannula. The incubation periods utilized in the experiment were 0, 4, 8, 16, 24, and 48 h, and a 4 × 2 × 2 split-plot design was employed. The results showed that the CP degradation fractions and CPD were significantly affected by forage family, species, and location of growth. Wide variations in degradation kinetics and degradability existed among and within the cereal and legume fodders, with wheat and jantar ranked at the top. Legume forages had larger soluble fractions, smaller potentially degradable fractions, and rapid rates and extent of degradation of dietary proteins than cereal forages. The cooler climatic conditions at location 2 increased the rapidly degradable protein fraction and overall CPD, whereas the warmer climatic conditions enhanced the slowly degradable protein fractions, thereby reducing the overall protein degradability in tropical forages. It was quite evident that some fodder species, such as maize among the cereal fodders and mustard among the legume fodders, remained quite non-responsive to the effects of the climatic conditions. A moderately positive and linear relationship between the rate of degradation and CPD was established for cereals, whereas a strongly positive and quadratic relationship was established for legume fodders. In conclusion, forage species, family, and location of growth significantly affected the degradation fractions and degradability of tropical cereal and legume fodders. Full article

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. Full article

This study was designed to explore the potential of infrared thermography (IRT) as an alternate approach for early pregnancy diagnosis in buffaloes. The surface temperature (ST) of different regions (eyes, muzzle, flanks, and vulva) was determined in 27 buffaloes using IRT from the day of artificial insemination (AI; Day 0), and measurement was repeated every fourth day until Day 24 post-AI. From all regions, the ST in each thermograph was recorded at three temperature values (maximum, average, minimum). Pregnancy status was confirmed through ultrasonography on Day 30, and animals were retrospectively grouped as pregnant or non-pregnant for analysis of thermographic data. In pregnant buffaloes, all three values of ST were significantly greater (p ≤ 0.05) for the left flank, while, in the left eye and vulva, only the maximum and average values were significantly greater. By contrast, the maximum ST of the muzzle was significantly lower (p ≤ 0.05) in pregnant buffaloes compared to non-pregnant buffaloes. However, the ST of the right eye and right flank did not show significant temperature variation at any value. These findings suggest that IRT has the potential to identify thermal changes associated with pregnancy in buffaloes at an early stage. Full article

Fodder scarcity, inadequate nutritional quality, and lack of degradation kinetics research are among the serious concerns hindering sustainable development of livestock globally. Rumen degradation kinetics data on neutral detergent fibre (NDF) in buffaloes are lacking for most tropical forage species. This study evaluated the effect of forage species, family, and growing location on NDF concentration and in situ degradability of six tropical cereal and four legume fodder crops. The fodder crops were grown following uniform recommended agronomic practices at three different agroecological locations and harvested at the appropriate growth stage: cereals at booting and legumes at 50% flowering. Later, the dried ground forage samples were incubated in the four rumen-cannulated Nili-Ravi buffalo cows in a four × two × three split-plot design for 0, 4, 8, 16, 24, 48, 96, and 168 h. The degradation fractions and degradability, expressed either on an NDF or dry matter basis, were considerably affected by forage family (cereal vs. legume) and growing locations and their interaction. Legume fodders degraded more rapidly but to a lesser extent than cereal fodders. The chemical components, notably the NDF, showed a significant but moderate negative relationship with the effective NDF degradability. Among studied fodders, the legumes had a significantly lower NDF concentration and subsequent degradability than the cereals. Although the cereals showed a slower rate of NDF degradation, their overall degradability was higher. The agro-climatic variability among three locations strongly impacted the NDF concentrations and fractions in the tropical forages. Effective NDF degradability was also correlated with the fodders’ NDF concentration, especially in cereals where the nature of the correlation was negative. In conclusion, the nutritional composition and NDF degradation parameters of the fodders were significantly affected by the species, family, and location of growth and their interactions. These results will help to improve agronomy and usage of these important fodder crops. Full article

The current study aimed to evaluate the impact of increasing cooling sessions from three to five times a day on milk yield and the welfare of dairy buffaloes during a semiarid summer in Pakistan. Eighteen Nili Ravi buffaloes were randomly assigned to three cooling strategies: (1) CTL, where buffaloes were cooled with a handheld hosepipe twice daily for 5 min each; (2) 3CS, where buffaloes were cooled using sprinklers three times daily; and (3) 5CS, where buffaloes were cooled using sprinklers five times daily. Each sprinkler cooling session lasted 1 h, with a 6 min cycle of 3 min of water on and 3 min off. Results showed that the 5CS group produced 1.6 and 3.2 kg more milk per day compared to the 3CS and CTL groups, respectively (p < 0.001). Both the 5CS and 3CS groups had consistently lower core body temperatures and respiration rates than the CTL group. Buffaloes in the 5CS group spent significantly more time eating (p < 0.001). Additionally, the 5CS group exhibited lower cortisol and blood urea nitrogen levels (p = 0.001) and higher glucose levels than the CTL group (p = 0.006). In conclusion, increasing cooling sessions to five times daily improved milk yield and welfare compared to the traditional cooling strategy (CTL) in dairy buffaloes during semiarid summers, highlighting the benefits of optimized cooling practices. Full article

Domestic buffaloes (Bubalus bubalis), known as water buffaloes, play a key role as versatile multipurpose agricultural animals in the Asiatic region. Pakistan, with the second-largest buffalo population in the world, holds a rich domestication history of buffaloes. The overall trends in buffalo production demand the genomic characterization of Pakistani buffalo breeds. To this end, the resequencing data of Pakistani breeds, along with buffalo breeds from 13 other countries, were retrieved from our previous study. This dataset, which contained 34,671,886 single-nucleotide polymorphisms (SNPs), was analyzed through a pipeline that was developed to compare possible allele differences among breeds at each SNP position. In contrast, other available tools only check for positional SNP differences for breed-specific markers. In total, 1918, 1549, 404, and 341 breed-specific markers were identified to characterize the Nili, Nili–Ravi, Azakheli, and Kundi breeds of Pakistani buffalo, respectively. Sufficient evidence in the form of phenotypic data, principal component analysis, admixture analysis, and linkage analysis showed that the Nili breed has maintained its distinct breed status despite sharing a close evolutionary relationship with the Nili–Ravi breed of buffalo. In this era of genome science, the conservation of these breeds and the further validation of the given selection markers in larger populations is a pressing need. Full article

The Nili Ravi, a buffalo breed from Pakistan, significantly contributes to the dairy industry. Mastitis is one of the major challenges affecting milk production in this breed. The objective of the current study was to identify the bacterial communities and diversity in healthy and mastitic milk of this breed. Milk samples (n = 14) were collected from Nili Ravi buffaloes with different udder health statuses, i.e., healthy (5), subclinical mastitis (4), and clinical mastitis (5). The DNAs were extracted, subjected to partial amplification of 16S rDNA (V3 and V4 regions), and sequenced using the Illumina platform. The results revealed variations in the bacterial communities in the milk of animals with different udder health statuses. Proteobacteria was the predominant phylum in the healthy group, while clinical and subclinical mastitis milk had a higher abundance of Firmicutes. Dominant bacterial genera in the healthy group were Streptococcus (11.60%), Herbaspirillum (7.65%), and Staphylococcus (4.70%), whereas the clinical mastitis group was dominated by Streptococcus (33.96%), Staphylococcus (7.87%), and Corynebacterium (2.68%), and the subclinical mastitis group was dominated by Bacillus (15.70%), Corynebacterium (6.70%), and Staphylococcus (6.58%). Assignment of operational taxonomic units at the species level resulted in most species being assigned to uncultured or unknown bacteria or remaining unassigned. Alpha diversity indices indicated lower microbial diversity in the clinical mastitis group, while beta diversity indices showed a scattered pattern of sample clustering in PCA plots among different groups. It is concluded that bacterial diversity in the milk of Nili Ravi buffaloes suffering from clinical mastitis is lower compared to healthy and subclinical mastitis cases. It is concluded that the variations in the microbiota of healthy and mastitic milk may be further investigated and exploited as signature microbes associated with the udder health status of Nili Ravi buffalo. Full article

Buffalo is the second source of milk in the world, and its milk is rich in nutritive components. It is well-known that breed influences milk composition. This work aimed to compare the detailed milk composition of three buffalo breeds (Murrah, Nili-Ravi, and Mediterranean) housed under the same environmental conditions. Mediterranean buffalo milk showed a significantly higher content of fat, protein, and some fatty acids. Moreover, the milk from the Mediterranean breed was characterized by the highest content of sphingomyelin (SM), cholesterol, and lanosterol. However, the Murrah buffalo milk contained the highest amount of total unsaturated fatty acids, phosphatidylinositol, and whey proteins. Furthermore, the Nili-Ravi buffalo milk was characterized by the highest content of total saturated fatty acids, phosphatidylglycerol, squalene, lathosterol, stigmasterol, beta-sitosterol, and casein fractions. Nevertheless, the lactose and amino acid profiles of the milk remained almost similar across the three buffalo breeds. The generated results in this study enable a comprehensive understanding of the milk constituent variability that is linked to buffalo breeds, which may support the acquirement of essential scientific knowledge on milk ingredient–processing interactions that will offer a foundation of knowledge for Chinese dairy processors in terms of milk processability and innovation. Full article

This article focuses on the toxic element chromium (Cr) in wastewater, its incorporation into soil plant systems, and its relevant toxicity in the food chain as assessed by a health risk assessment from dietary intake. The Nili Ravi buffalo is an important cattle inhabiting Sargodha, Punjab, Pakistan, and forage crops grown on soils contaminated with Cr might cause toxicity in the food chain by local inhabitants eating meat. The soil, forage and animal blood samples were collected from five different locations in Tehsil Sahiwal (Chak Dhool, Bagabalocha, Chandia, Dhool Bala and Kakrani) twice at six-month intervals. A total of 30 samples from each ecological zone were collected from the soil and forage crops (Zea mays, Sorghum bicolor, Trifolium alexandrinum). The samples from zone-V and zone-IV showed the maximum concentration of Cr because these areas receive highly contaminated water for irrigation. The Cr was greater than the permissible limits. Environmental indices for all samples ranged below 1. The bioaccumulation and pollution load of Cr in soil and forage crops due to wastewater irrigation can contaminate the whole food chain via the soil, forages and animals. The health risk index (HRI) and a high value of enrichment factor were found for Cr in some sites. The Cr concentration was higher during the summer season than winter. Fodder crops with different concentrations and an elevated level of Cr were observed in maize. Attention should be paid when wastewater is used for fodder crop irrigation and its potential risks to human health following dairy product (milk, meat) entry into the food chain. Full article