Nili-Ravi Buffalo Research Articles

Chromium Poisoning in Buffaloes in the Vicinity of Contaminated Pastureland, Punjab, Pakistan

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.

Effect of body condition score at calving on transition success in Nili Ravi buffaloes

Body condition score (BCS) at calving is a vital indicator of the effectiveness of the beginning of lactation in dairy animals. The purpose of this study was to examine the effect of BCS at calving on milk production and transition success in dairy buffaloes. Thirty-six (36) Nili Ravi buffaloes were enrolled at 40 days of expected calving and followed through 90 days of lactation. The buffaloes were categorized into three groups according to their BCS (on a scale of 1-5 with 0.25 increments) as follows: 1) low, buffaloes with BCS ≤ 3.0; 2) medium, buffaloes with BCS 3.25-3.5; and 3) high, buffaloes with BCS ≥ 3.75. All buffaloes were fed a similar diet ad libitum. The lactation diet had increased concentrate allowance according to milk yield. The results revealed that the BCS at calving did not affect milk yield; however, fat percentage (fat%) was lower in the low-BCS group. Dry matter intake (DMI) was similar among the treatment groups, although post-calving BCS loss was greater in the high-BCS group compared to the medium- and the low-BCS groups. Similarly, the buffaloes in the high-BCS group had higher non-esterified fatty acids (NEFA) concentration compared to the low- and medium-BCS groups. No cases of metabolic disorders were observed during the study. The present results suggest that the buffaloes in the medium-BCS group appeared to perform better compared to the low- and the high-BCS groups with respect to milk fat% and blood NEFA concentration.

Variations of Raw Milk Components and Amino Acid Profiles in Different Dairy Buffalo Crossbreds

This study aimed to assess the relationship between milk components and amino acid (AA) profiles among different buffalo crossbreds in Bangladesh. A total of thirty-six (36) lactating buffaloes were selected from Murrah, Nili-Ravi, and Mehsana crossbreds, and they were assigned to 03 groups, each with 12 buffaloes. The total experimental period was 10 weeks, including the initial 10 days of diet adjustment. The results from the experiment revealed that milk protein, fat, and total solids contents of Murrah crossbred was significantly (p<0.05) higher than Mehsana and Nili-Ravi crossbreds. In contrast, the lactose content of all buffalo crossbreds was statistically similar (p>0.05). In the case of milk amino acid contents, all the milk samples entailed a higher concentration of Glutamic acid (0.9-1.00 g/100 g of milk), whereas Cysteine had the lowest concentration (0.02-0.05 g/100 g). The most prevalent essential amino acids were Leucine, Lysine, and Phenylalanine, whereas the most prevalent non-essential amino acids were Glutamic acid, Proline, Asparagine, and Serine. Lysine, Isoleucine, Leucine, Phenylalanine, Cysteine, and Histidine concentrations differed significantly (p<0.05), and at the same time as the other 11 AAs concentrations were found non-significant (p>0.05). The highest Lysine: Methionine ratio observed in Murrah buffalo was about 3.20%, while in Mehsana and Nili-Ravi buffalo was about 2.80 and 2.50%, respectively. Thus, it was evident from the study that the raw milk components and amino acid composition vary considerably in the different lactating buffalo crossbreds. Finally, the compositional data of raw milk may create the way of sustainable use of milk from dairy buffalo crossbreds and improve food and nutrition security, particularly in developing countries.

Identification of differential methylome signatures of white pigmented skin patches in Nili Ravi buffalo of India.

The DNA methylation events mark a major epigenetic change in the genome, reflecting non-genetic disease developments and varied phenotypes. The water buffalo is a dairy production animal with wide agro-climatic distribution in India. Breed-wise the coat color of water buffalo varies from ash-gray to jet black. A typical pigmentation pattern is found in one of the breeds of North India, Nili Ravi, with variedly distributed white patches. The DNA methylation pattern could potentially reveal the epigenetic factors responsible for the pigmentation patterns. To address this question, the DNA isolated from the skin tissues of Nili Ravi with varied white pigmentation and black Murrah buffaloes was subjected to reduced representation bisulfite sequencing. DNA methylation analysis revealed, 68.44%, 63.39%, and 47.94% of the promoter regions were hypermethylated in Nili Ravi over-white versus Murrah, Nili Ravi under-white versus Murrah, and Nili Ravi under-white versus Nili Ravi over-white, respectively. Major genes identified to be differentially methylated among over-white and under-white skin tissues in Nili Ravi included TBX2, SNAI2, HERC2, and CITED1. Overall the results have indicated differential methylation patterns to be potentially involved in hyper or hypopigmentation in Nili Ravi and Murrah buffaloes.

Effects of maize silage substitution with sugarbeet or citrus pulp ensiled with corncobs on growth performance, digestibility, and economic benefits in buffalo calves.

Ensiling of high-moisture agro-industrial wastes with dry roughage as animal feeds may be a secure way to reduce the feeding cost. The objective of this study was to investigate the effects of either sugarbeet or citrus pulp ensiled with ground corncobs and used as a replacement of maize silage or maize fodder in total mixed ration (TMR) for fattening of buffalo calves. Twenty male calves of Nili-Ravi buffalo (aged 14 ± 2months weighing 133 ± 5kg) were randomly assignedto one of four iso-nutritious (12% CP and 69% TDN) experimental diets with 50:50 forage to concentrate ratio (dry matter basis) as: (1) TMR maize silage, (2) TMR sugarbeet pulp ensiled with corncobs, (3) TMR citrus pulp ensiled with corncobs, and (4) TMR green maize fodder with wheat straw. All calves received TMRs for ad libitum intake twice daily for 80-day period. Daily feed intake, fortnightly body weights, and digestibility and N balance study during last 15days were recorded of individual animals. The calves fed TMR sugarbeet pulp silage got highest daily gain (856g/d, P < 0.05) than citrus pulp silage (776g/d) or green maize fodder (704g/d). Average intake of nutrients was same among all the calves. Feed conversion ratio and digestibility of DM, NDF, and ADF were higher (P < 0.05) with TMR sugarbeet pulp silage compared to other TMRs. Economic benefit (output/input) with TMR sugarbeet pulp silage had 15.46, 34.27, and 37.36% higher effect for fattening of buffalo calves compared to citrus pulp silage, green maize fodder, and maize silage-based TMRs, respectively. In conclusion, sugarbeet pulp ensiled with ground corncobs is simple and appropriate method for silage making and its inclusion up to 50% of DM in TMR had pronounced economic impact to the production of buffalo calves.

Genetic variants of CSN1S1, CSN2, CSN3, and BLG genes and their association with dairy production traits in Sahiwal cattle and Nili-Ravi buffaloes

Milk protein genes are associated with milk yield and composition in dairy animals. The present study aimed to identify milk protein genes (CSN1S1, CSN2, CSN3, and BLG) genetic variants and their association with milk yield in Sahiwal cattle and Nili-Ravi buffaloes. One hundred animals from each species were selected to collect blood samples and milk production records. Primers were designed for these milk protein genes for PCR amplification. Sequencing of resultant PCR products revealed a higher number of SNPs (13 vs. 7, 5 vs. 1, and 6 vs. 2) in Sahiwal as compared to Nili-Ravi animals in CSN1S1, CSN2, and CSN3 genes, respectively. However, a single SNP was observed in BLG gene of both species. Association analysis revealed that one SNP in BLG gene of Nili-Ravi was associated (p < 0.05) with 305-day milk yield. Two SNPs at CSN1S1 gene in Sahiwal were associated with dry-period. Similarly, one SNP at CSN1S1 and two SNPs at CSN3 gene showed significant association (p < 0.05) with average calving-interval in Sahiwal while two SNPs in CSN1S1 gene were associated (p < 0.05) with this trait in Nili-Ravi. These SNPs could be helpful as candidate variants for marker-assisted selection in cattle and buffaloes for improvement of lactation performance.

Comparative efficacy of coconut water diluent with different semen extenders for cryopreservation of Nili Ravi buffalo bull semen

In this study, efforts were made to investigate fresh semen parameters and to select a suitable extender for buffalo semen cryopreservation. Experiment I, fresh undiluted seminal parameters were determined while Experiment II, efficacy comparison of coconut water extender (CWE) with tris citric acid extender (TCAE) and skimmed milk extender (SME) was made. Four bulls single ejaculate was collected weekly for 5 and 10 weeks for Experiment I and II, respectively however osmotic pressure replicates were 16 and 6 for semen and seminal plasma, respectively. In Experiment I, each bull spermatozoa concentration, motility (%), semen volume and pooled semen percentage NAR, PMI, viability and MTT (3-(4, 5-dimethyl thiazol-2-yl)-2,5-diphenyl tetrazolium bromide) reduction rate was checked. In Experiment II, pooled semen added to extenders and then equilibrated (4oC) and filled to obtain 20×106 spermatozoa/0.5 ml straws before plunging in liquid nitrogen. Percentage thawed spermatozoa viability, normal acrosomal ridge (NAR), motility, DNA damage, plasma membrane integrity (PMI) and lipid peroxidation (nM) recorded. In Experiment I, seminal parameters as spermatozoa concentration (1226.43±71.48 million/mL), semen volume (2.84±0.14 mL), viability (90.05±0.71%), motility (77.13±0.71%), PMI (86.23±0.34%), NAR (94.67±0.30%) and MTT reduction rate (0.290±0.06) while osmotic pressure of seminal plasma (294.83±3.87 mOsm/kg) and semen (290.87±2.58 mOsm/kg) was recorded. In Experiment II, TCAE higher (P&lt;0.05) sperm motility noted compared to CWE and SME whereas percentage viability, NAR, PMI, DNA damage was non-significant. Lipid peroxidation compared to SME and TCAE was higher (P&lt;0.05) in CWE. In conclusion, based on sperm motility and lipid per oxidation, TCAE was more efficient for cryopreservation of buffalo semen.

Effect of Parity, Lactation Length and Season of Calving on Milk Production in Nili Ravi Buffaloes Maintained at Livestock Farm, GADVASU, Ludhiana.

Background: Nili Ravi buffalo breed is one of the best milk-producing buffalo breeds with excellent potential for milk production. In the present investigation, we studied the effect of non-genetic factors on milk production traits in Nili Ravi buffalo. Methods: In the present study, we analyzed the factors affecting milk production traits in 279 standardized lactation records in Nili Ravi buffaloes maintained at the Directorate of Livestock Farm, GADVASU, Ludhiana, from 2009 to 2020. The effect of lactation length (LL), parity and season of calving (SOC) on milk production were analyzed using GLM (General Linear Model) IBM SPSS Statistics 26 version software. Result: The overall 305 days corrected milk yield of Nili Ravi buffalo was 2247.09±26.51 kg with an average lactation length of 274.72±62.78 days. The effect of SOC and parity number was found to be significant (P≤0.05) on the 305 days of corrected milk yield, whereas the effect of lactation length was found significant on lactation milk yield (P≤0.01). It is concluded that the lactation length, parity and calving season significantly affect the milk production trait in Nili Ravi buffalo.

Kisspeptin-10 in cryodiluent improves the post-thaw quality of Nili-Ravi buffalo (Bubalus bubalis) bull spermatozoa.

Effects of kisspeptin-10 as antioxidant in cryodiluent were evaluated on post-thaw quality of buffalo spermatozoa. Qualified semen samples from five bulls were pooled, divided into five aliquots and extended in Tris-citric acid cryodiluent containing differential doses of kisspeptin-10 (5, 10, 15, and 20 μmol L-1 and negative control. Extended sperm suspension was cooled to 4°C, packaged in 0.54 ml straws and cryopreserved. At post-thawing, catalase (unit mg-1 ), peroxidase (unit mg-1 ) and reduced glutathione (μmol L-1 ) levels were highest (p < 0.05) with 20 μmol L-1 of kisspeptin-10 as compared to negative control. Moreover, lipid peroxidation (nmol L-1 min-1 mg protein-1 ) level was lowest (p < 0.05) with 20 μmol L-1 of kisspeptin-10. Sperm progressive motility (%), rapid velocity (%) and kinematics were higher (p < 0.05) with 15 and 20 μmol L-1 of kisspeptin-10 as compared to negative control. Supra-vital plasma membrane integrity (%), viable sperm with intact acrosome (%) and DNA integrity (%) were improved (p < 0.05) with all doses of kisspeptin-10 as compared to negative control. It was concluded that the addition of 15 and 20 μmol L-1 kisspeptin-10 in cryodiluent ameliorated the overall frozen-thawed quality parameters of Nili-Ravi buffalo spermatozoa.

Study on Various Luteal Characteristics Using Doppler Ultrasonography for Early Pregnancy Diagnosis in Nili-Ravi Buffaloes.

The objective of current study was to assess the trend in various luteal characteristics viz luteal size (LS), plasma progesterone (P4) concentration, and luteal blood flow (LBF) using color Doppler imaging (CDI) and power Doppler imaging (PDI) modes in pregnant and nonpregnant Nili-Ravi buffaloes. Lactating, cyclic, and healthy Nili-Ravi buffaloes (n = 09) without any reproductive abnormality were selected in present study. Buffaloes were synchronized using Ov-Synch, and fixed-time artificially insemination was performed (day = 0). Pregnancy was diagnosed on 30-day post-AI using B-mode ultrasonography based on presence or absence of embryonic heartbeat. Ovaries of all animals were scanned from day 5 till 21 post-AI using both B-mode and Doppler ultrasonography to measure LS and LBF. After each ovarian ultrasound examination, blood samples were collected via jugular venipuncture to determine plasma P4 concentration. According to results, LBF using CDI and PDI was significantly higher (P ≤ 0.05) in pregnant buffaloes on days 13 and 15 post-AI, respectively. The mean LS and plasma P4 concentration did not differ (P ≥ 0.05) between pregnant and nonpregnant animals until day 15 post-AI. However, a significant difference (P ≤ 0.05) was noticed for both on day 17 and onwards. It is concluded that LBF is a more sensitive luteal character as compared to LS and P4 for earlier pregnancy diagnosis in Nili-Ravi buffaloes when ascertained through CDI.

Genomic analysis of arginine vasopressin gene in riverine buffalo reveals its potential association with silent estrus behavior.

Poor estrus expression behavior causes suboptimal reproductive efficiency through poor conception rate. Various signaling pathways are involved in estrus expression but arginine vasopressin (AVP) gene with oxytocin predominantly regulates estrus behavior. This study aimed to perform genomic characterization and evolutionary dynamics of AVP gene through association testing of the novel polymorphic loci and comparative genomic analysis to explore the potential effect of AVP gene on estrus behavior of Nili-Ravi buffaloes. 198 Nili-Ravi buffaloes were screened for the quest of novel polymorphism in the AVP gene. In exon-1, five polymorphic sites were detected including deletion of two (c.47delA and c.57delA) nucleotides that caused drastic variation in subsequent amino acid sequence due to frame shift including functional short peptide of nine residues. The 3-D structure revealed a loss of transmembrane loop between 16 and 31 residues in Nili-Ravi buffalo AVP protein sequence, suggesting that missing loop apparently reduced the gene functionality in Nili-Ravi buffalo by inhibiting cellular reactions and muting the animal estrus cyclicity. Three polymorphisms detected in AVP gene were significantly associated with silent estrus (P <0.05). The comparative genomic analysis revealed that AVP gene is present on chromosome 14 having one conserved motif (Neurohypophysial) in buffalo. This study suggested the potential use of polymorphic sites as promising genetic markers for selection of buffaloes with pronounced estrus expression.

Validation of pregnancy associated glycoproteins-based ELISA kits to determine early pregnancy status in lactating Nili-Ravi buffaloes.

The objective of the current study was to evaluate pregnancy-associated glycoproteins (PAGs) based enzyme-linked immunosorbent assay (ELISA) kits utilizing whole blood, serum, or milk samples for diagnosis of early pregnancy status in lactating Nili-Ravi buffaloes. Dairy buffaloes (n =174) of mixed parity, 4-6 years of age, having mean (± standard deviation) days in milk 165 ± 87, and body condition score of 3.26 ± 0.34 were randomly enrolled in this study. Buffaloes were exposed to penile deviated bulls with 12 h interval for estrus detection during peak breeding season and eventually bred naturally at their respective standing estrus (day 0). Blood and milk samples were collected at days 24, 28, and 35 post-breeding to run a rapid visual pregnancy test® (RVPT) as a buffalo-side test or ELISA-based assay in the laboratory to detect early pregnancy status. Transrectal B-mode ultrasonography was performed to diagnose pregnancy at day 35 post-breeding and used as a gold standard to validate results of RVPT or ELISA-based tests. The RVPT is a visual readout test for pregnancy detection and had sensitivity (77.9 vs. 89.7 vs. 93.3%), specificity (77.9 vs. 89.7 vs. 93.3%), and accuracy (84.5 vs. 90.1 vs. 94.2%) at days 24, 28, and 35 post-breeding, respectively. The PAGs were assayed using ELISA kits in serum and had sensitivity (77.9 vs. 89.7 vs. 93.3%), specificity (84.2 vs. 87.7 vs. 93.9%), and accuracy (82.1 vs. 88.4 vs. 93.7%) at days 24, 28, and 35 post-breeding, respectively. Similarly, PAGs were also analysed using ELISA kits in milk samples and had sensitivity (77.6 vs 89.5 vs 95.0%), specificity (89.1 vs 91.9, vs 93.9%), and accuracy (85.1 vs 91.1 vs 94.3%) at days 24, 28, and 35 post-breeding, respectively. Overall, the Kappa values in this study exceeded 0.85 at day 35 post-breeding using RVPT or ELISA-based test kits in serum or milk samples, indicating a high level of agreement between PAGs detection method and gold standard for pregnancy diagnosis. The pregnancy outcomes based on ELISA-based PAGs detection at day ≥28 post-breeding had a high negative predictive value, indicating that the probability of incorrectly administering prostaglandins to pregnant buffaloes would be low if these tests were implemented on a commercial dairy herd. Taken together, it is concluded that PAGs-based determination of pregnancy using RVPT or ELISA in blood, serum, or milk samples can be used effectively for pregnancy diagnosis at ≥28 days post-breeding with more than 90% accuracy in Nili-Ravi buffaloes.

Effect of Prepartum Dietary Energy Level on Production and Reproduction in Nili Ravi Buffaloes.

Simple SummaryThe objective was to evaluate the effect of prepartum dietary energy level on performance of Nili Ravi buffaloes. Buffaloes were fed low, medium or high energy diet during 63 days prepartum and same lactation diet during 14 weeks postpartum. Buffaloes fed the high energy diet had better performance compared with buffaloes fed the medium or low energy diet. While, prepartum feeding of low energy diet adversely affected buffalo performance.The objective of this study was to evaluate the effect of prepartum dietary energy level on postpartum production and reproduction in Nili Ravi buffaloes (n = 21). The buffaloes were offered low energy (LE: 1.31 Mcal/kg DM NEL (net energy for lactation)), medium energy (ME: 1.42 Mcal/kg DM NEL) or high energy (HE: 1.54 Mcal/kg DM NEL) diet for 63 days prepartum, and received the same lactation diet (LD: 1.22 Mcal/kg DM NEL) during 14 weeks postpartum. The effects of dietary energy level and week were analyzed with Proc GLIMMIX model. Dry matter intake (DMI) was lower in buffaloes fed the LE diet compared with buffaloes fed the ME or HE diet. Calf birth weight (CBW) was higher in buffaloes fed the HE diet compared with buffaloes fed the ME or LE diet. Milk production was similar in buffaloes fed the HE, ME or LE diet within 14 weeks postpartum and throughout the lactation. Milk fat was higher in buffaloes fed the LE diet compared with buffaloes fed the ME or HE diet. Milk protein and lactose yields was high in buffaloes fed the HE diet compared with buffaloes fed the ME or LE diet. Body condition score was high in HE and was affected by diet × week interactions during pre and postpartum period. The concentrations of β-hydroxybutyrate (BHBA) and triglycerides in serum was lowest in buffaloes fed the HE diet compared with the buffaloes fed the ME or LE diet. The buffaloes fed the HE diet had early uterine involution (UI), first estrus, short dry days, and calving interval (CI) compared with buffaloes fed the ME or LE diet. None of buffaloes fed the LE diet exhibited estrus during the first 14 weeks postpartum compared with buffaloes fed the ME or HE diet. In conclusion, prepartum feeding of high energy diet can be helpful in improving the postpartum productive and reproductive performance in Nili Ravi buffaloes.

Sprinkler flow rate affects physiological, behavioural and production responses of Holstein cows during heat stress

Holstein Friesian cows are known for their high milk yield. They are popular in the commercial dairy sector of Pakistan. Heat stress is a major challenge for these animals owing to the longer summer season with high ambient temperatures (35 - 40 °C) and relative humidity (30 - 50%) in the area (Butt et al., 2020). Mitigating the effects of heat stress requires substantial investments in energy and water. Spraying dairy cows with water in the holding pen or at the feed bunk is common practice as it lowers body temperature and respiration rate (Kendall et al., 2007; Chen et al., 2013; 2016), and improves feed intake and milk yield (Keister et al., 2002). Owing to the recent decrease in global groundwater, studies have focused on strategies to reduce the use of water to cool dairy cows (Chen et al., 2016). Sprinkler flow rates of 1.3 and 4.9 L/min had similar effects on physiological responses of cows to heat load but produced a 73% difference in water use (Chen et al., 2016). Similarly, Tresoldi et al. (2019) concluded that a sprinkler flow rate of 4.9 L/min had poor efficiency compared with 3.3 L/min, as the additional water did not result in increased biologically relevant cooling. These studies were conducted in Mediterranean climates with an average temperature humidity index in the range of 76 to 78. A recent study with Nili Ravi buffaloes revealed a sprinkler flow rate 1.25 L/min was more efficient than one of 2.0 L/min during subtropical summers (Bah et al., 2021). Because water is projected to be scarce in Pakistan by 2030 (Mustafa et al., 2013), holistic strategies for water conservation in the dairy sector are important nationally. The objective of this study was to evaluate the effects of various sprinkler flow rates on the performance of lactating Holstein Friesian cows in a typical corporate dairy farm setting during the subtropical summer. Performance of these cows was assumed to be similar, irrespective of the sprinkler flow rates.

COMPARATIVE STUDY OF FEEDING MILK REPLACER, WHOLE MILK PLUS MILK REPLACER BLEND AND WHOLE MILK ON DRY MATTER INTAKE, AVERAGE DAILY GAIN AND FEED CONVERSION RATIO IN NILI RAVI BUFFALO CALVES

COMPARATIVE STUDY OF FEEDING MILK REPLACER, WHOLE MILK PLUS MILK REPLACER BLEND AND WHOLE MILK ON DRY MATTER INTAKE, AVERAGE DAILY GAIN AND FEED CONVERSION RATIO IN NILI RAVI BUFFALO CALVES

Pregnancy and calving rates improved using modified swim-up method for buffalo semen sexing.

Present study aimed to evaluate field fertility rate and calf sex ratio of Nili Ravi buffalo semen sexed through modified swim-up method (Animal Reproduction Science, 182, 2017, 69). For this purpose, five mature Nili-Ravi buffalo bulls kept at semen production unit, Qadirabad, Pakistan, were selected. Two consecutive ejaculates per week were collected with artificial vaginą for 3weeks. Qualified semen ejaculates were pooled and divided into two aliquots. The first aliquot was processed by routine procedure (control), whereas the second was processed by modified swim-up technique. After separation, semen was diluted in tris-citric acid extender and cryopreserved using standard techniques. Sexed semen was evaluated for fertility trials during peak breeding season. Artificially inseminated animals were examined for pregnancy rate through rectal palpation at least 3months after insemination under field conditions. Calving ratio of female and male calves were recoded after Parturition. The fertility rate was higher (p<.05) in X-sorted sperm (70%) as compared with control (47%). The female calf ratio was higher (p<.05) in X-sorted sperm (78.58%) compared with control (53.3%). In Conclusion, conception rate and production of female calf were significantly higher with sexed semen separated through modified swim-up method compared with unsexed control.

Performance and welfare of dairy buffaloes subjected to different cooling strategies during subtropical summer.

This study aimed to evaluate the effects of different cooling strategies on milk yield and physiological and behavioral responses of Nili Ravi buffaloes during subtropical summer in Pakistan. Thirty Nili Ravi buffaloes were randomly assigned to three treatments: (1) CNT buffaloes cooled with the application of water using handheld hosepipe twice daily, 3min each with a water flow rate of 40 L/min; (2) 2SS, buffaloes cooled with sprinklers twice daily; (3) 3SS, buffaloes cooled with sprinklers thrice daily. Each of the sprinkler sessions lasted for 1h with a 12-min cycle (3min water on, 9min off). The trial was carried out from mid-July until the end of September 2019. The average ambient afternoon temperature humidity index was 86.9. The results indicated that the buffaloes in the 3SS group had lower respiration rate and rectal temperature than those in the CNT and the 2SS groups (P < 0.01). The 3SS group had more daily milk yield (P = 0.019) and milk fat % (P < 0.01) than the CNT and 2SS groups. The lying time and the lying bout length were significantly longer in the 3SS than in the CNT and 2SS groups. The blood cortisol levels tended to be lower in the 3SS group than in the CNT and 2SS groups (P = 0.051). In conclusion, the 3SS cooling strategy had more milk yield, more milk fat, and better welfare than the CNT strategy using less groundwater and both performed better than the 2SS strategy.

First substantiation of clinico-molecular investigation of pathogenic Listeria monocytogenes in Nili-Ravi buffaloes.

Listeriosis is a zoonotic disease of humans, animals, birds, fish, and crustaceans worldwide. Domestic animals, especially ruminants, are more susceptible to listeriosis. This infectious disease is caused by Listeria monocytogenes, an intracellular bacterium that can cross blood-brain, placental and intestinal barriers. In Pakistan, the incidence and reliable diagnostic tools for the L. monocytogenes are unidentified in Nili-Ravi buffaloes. This study was designed to inspect listeriosis in buffaloes through molecular techniques and haemato-biochemical analyses. A total of 230 samples (115 milk and 115 faecal samples) were collected from symptomatic listeriosis cases in Nili-Ravi buffaloes of 3 geographical districts (Rawalpindi, Faisalabad, and Muzaffargarh) Punjab, Pakistan. These samples were processed for DNA extraction using commercialized kits, and L. monocytogenes was confirmed by conventional PCR. The results revealed that 6.08% and 4.34% of the isolates from milk and faecal samples were found positive for L. monocytogenes, respectively. The phylogenetic analysis of these isolates showed 97-100% similarity to isolates from the USA, Switzerland, Japan, and India. The accession numbers on NCBI GenBank appeared as HF558398 (Switzerland), KP965732 (India), EU372032 (USA), and LC259850 (Japan). Haemato-biochemical examinations showed that the values of WBCs, plasma fibrinogen, ALT, and AST significantly increased (P<0.05) in diseased buffaloes compared to healthy ones. The occurrence of listeriosis in buffaloes urges continuous monitoring and surveillance to prevent this emerging disease in Pakistan.

Cryopreservation of Nili-Ravi buffalo bull semen: yearly postthaw analyses to depict climatic effects

The present study was aimed to observe the postthaw quality of Nili-Ravi buffalo bull semen cryopreserved during heat stress (hot summer months) and normal climatic conditions (fall, winter, and spring months). For this purpose, four (n = 4) adult regular donor Nili-Ravi buffalo bulls were selected to collect and cryopreserve their semen throughout the period of an entire year. The cryopreservation of semen was done following standard freezing protocol. The semen doses cryopreserved during heat stress and normal climatic condition were thawed to determine the sperm motility, motion kinetics, viability, plasma membrane integrity, acrosomal integrity, and morphological abnormalities (head, mid-piece, and tail). The results showed that sperm motility and sperm motion kinetics did not alter (p > 0.05) across heat stress and normal climatic period. The sperm head, mid-piece and tail abnormalities did not differ (p > 0.05); however, total sperm morphologic abnormalities were higher (p < 0.05) during heat stress period, whereas the lower (p < 0.05) sperm viability, sperm plasma membrane, and acrosomal integrity were observed during heat stress period. In conclusion, the total morphologic abnormalities, viability, and acrosomal and plasma membrane integrity of buffalo bull sperm cryopreserved during hot summer months are compromised due to heat stress.

Assessment of prenatal ovarian and serum estrogen concentration in Nili-Ravi buffalo fetus

Assessment of prenatal ovarian and serum estrogen concentration in Nili-Ravi buffalo fetus