Fertility In Buffaloes Research Articles

Exploring the Effects of Nitric Oxide and Reactive Oxygen Species on Buffalo Sperm Quality: A Comprehensive Review

Buffalo reproduction plays a vital role in the livestock industry, where sperm quality is a critical determinant of successful fertilization and overall fertility. Nitric oxide (NO) and reactive oxygen species (ROS) are key regulators of various physiological processes, including sperm function. However, their dual role in modulating sperm quality is complex, as both excessive and insufficient levels can lead to detrimental effects. This review explores the intricate balance between NO, ROS and sperm quality in buffalo, emphasizing the molecular mechanisms underlying these interactions. NO is a signaling molecule involved in the regulation of sperm motility, capacitation and acrosome reaction, but its overproduction can result in nitrosative stress, impairing sperm function. Similarly, ROS, at physiological levels, are essential for sperm capacitation and acrosome reaction, yet an imbalance towards oxidative stress can lead to lipid peroxidation, DNA fragmentation and reduced sperm viability. This review also delves into the role of exogenous NO donors, such as sodium nitroprusside (SNP), in modulating NO and ROS levels, and their consequent impact on sperm quality. The dual effects of NO and ROS on buffalo sperm highlight the importance of maintaining an optimal redox balance to preserve sperm integrity and functionality. In conclusion, understanding the molecular pathways through which NO and ROS influence sperm quality is crucial for developing targeted strategies to enhance reproductive outcomes in buffalo. Future research should focus on identifying biomarkers of oxidative stress and evaluating the potential of antioxidant therapies to mitigate the adverse effects of NO and ROS, ultimately improving buffalo fertility.

Effect of Adding Autologous Platelet-rich Plasma to the Freezing Extender on the Post-thaw Quality and In vitro Fertility of Buffalo (Bubalus Bubalis) Spermatozoa

Effect of Adding Autologous Platelet-rich Plasma to the Freezing Extender on the Post-thaw Quality and In vitro Fertility of Buffalo (Bubalus Bubalis) Spermatozoa

Coenzyme Q10 improves the quality and invitro fertility of post-thawed buffalo (Bubalus bubalis) semen via its antioxidative effect.

This study aimed to determine the effects of Coenzyme Q10 (CoQ10) in the freezing medium on functional and oxidative stress parameters and invitro fertilization (IVF) rate of buffalo sperm. Collected samples were relocated to the laboratory for initial evaluation, gentle dilution in extenders, cooling (4°C, 2 h), equilibration (4°C, 4 h), packaging (straws, 0.5 mL), programmable freezing, and thawing (37°C, 30 s). Statistical analysis depicted that adding CoQ10 (100 μM) in a freezing medium caused a significant augmentation in total motility (%), average path, and straight-line velocities (μm/sec) of buffalo sperm than control. Adding CoQ10 (100 μM) improved sperm progressive motility, rapid velocity, and functional parameters (%) compared to the control and 10 μM of CoQ10. Moreover, CoQ10 in a freezing medium caused a significant augmentation in seminal plasma catalase (U/mL) and glutathione reductase (GSH; nmol/109 ) at 100 μM than control and other treatments. CoQ10 inclusion (100 μM) ameliorates seminal plasma superoxide dismutase (U/mL), glutathione-S-transferase (GST; nmol/mL/min) fructose (μg/mL), and ATP (nmol/million) than control. Furthermore, CoQ10 at 100 μM improved seminal plasma glutathione peroxidase (μM) levels than control, 10 μM, and 20 μM. Lastly, hydrogen peroxide (H2 O2; nM) production was significantly lower at 100 μM than at control and 10 μM. CoQ10 (100 μM) caused a significant augmentation in the un-capacitated pattern followed by a reduction in the capacitated pattern, and apoptosis-like changes (%) than control, and other treatments, whereas viability was increased than control and other treatments. CoQ10 (100 μM) significantly improved the IVF rate in comparison with control, CoQ10 at 10 μM, and 20 μM groups. In conclusion, the addition of CoQ10 (100 μM) in the freezing medium can improve the quality and invitro fertility of post-thawed buffalo semen via its antioxidative effect. Further studies are needed to evaluate the effect of CoQ10 on the invivo fertility of buffalo bull semen.

Genetic parameters for reproductive traits in the Italian Mediterranean buffalo using milk yield as a correlated trait

Until now, the genetic evaluation of the Italian Mediterranean Buffalo has been mainly focused on production traits. However, female fertility affects the efficiency of the dairy industry as it is essential to maintain the profitability of dairy farms. Indeed, the estimation of its genetic component is crucial for its improvement. In this study, 3 measures of buffalo's fertility were analyzed: the age at first calving (AFC), the interval between first and second calving (CI1), and the interval between second and successive calvings (CI2_12). Milk yield at 270 d (MY270) was used as a correlated trait. First, genetic parameters were estimated using 7,915 buffalo cows with first calving from 1991 to 2018, then breeding values were calculated from 236,087 buffalo cows. Genetic parameters were estimated by Bayesian inference fitting a multiple-trait animal model using the GIBBS1F90 program while BLUPF90 was used for estimation of breeding value. The heritability and repeatability estimates of fertility traits were low. The genetic correlations among fertility traits ranged from 0.10 (AFC-CI1) to 0.92 (CI1-CI2_12). Genetic correlation between MY270 and fertility traits was unfavorable, ranging from 0.23 to 0.48. The results from this study can be used as a basis for the future genetic improvement of fertility traits in the Italian Mediterranean Buffaloes.

Sulforaphane inclusion in a freezing medium augments post-thaw motility, functional and biochemical features, and fertility potential of buffalo (Bubalus bubalis) spermatozoa

Sulforaphane is a natural and highly effective antioxidant safeguarding the reproductive system, and alleviate oxidative stress. This study was designed in order to elaborate L-sulforaphane effect on semen quality, biochemical parameters, and fertility of buffalo (Bubalus bubalis) spermatozoa. Semen was collected from five buffalo bull with artificial vagina (42 °C) three times and evaluated for volume, consistency (color), motility, and sperm concentration. After critical examination, semen was diluted (50 × 106 spermatozoa per ml, 37 °C) in extenders with (2 μM, 5 μM, 10 μM, and, 20 μM) or without (control) sulforaphane, cooled (from 37 to 4 °C), equilibrated (4 °C), filled (straws, 4 °C), and cryopreserved (LN2, −196 °C). Data analysis exhibited that sulforaphane addition in extender augments total motility (%, 10 μM, and 20 μM than control), progressive motility (%), and rapid velocity (%, 20 μM than control), and velocity parameters (average path velocity, μm/s, straight line velocity, μm/s and curved linear velocity, μm/s, 20 μM than control, and 2 μM). Moreover, sulforaphane augments functional features (membrane functionality, mitochondrial potential, and acrosome integrity) of buffalo sperm (20 μM than control). Sulforaphane preserves biochemical features of seminal plasma of buffalo i.e., Calcium (μM), and total antioxidant capacity (μM/L), followed by reduction in lactate dehydrogenase (IU/L), reactive oxygen species (104 RLU/20 min/ 25 million), and lipid peroxidation (μM/ml) in 20 μM than control. Lastly, sulforaphane augments fertility rate of buffalo sperm at 20 μM than control, and 2 μM. Conclusively the existing study revealed that adding L-sulforaphane (20 μM) in a freezing medium augments motilities, kinematics, functional parameters, and fertility rate of buffalo spermatozoa. Correspondingly, sperm favorable biochemical features were also augmented with sulforaphane followed by reduction in oxidative stress parameters. Further studies are highly recommended to define the particular mechanism of action of sulforaphane in augmenting buffalo post-thawed semen quality, and in vitro fertility potential.

Identification of protein candidates in spermatozoa of water buffalo (Bubalus bubalis) bulls helps in predicting their fertility status.

The water buffalo (Bubalus bubalis) is an indispensable part of the Indian dairy sector and in several instances, the farmers incur economic losses due to failed pregnancy after artificial insemination (AI). One of the key factors for the failure of conception is the use of semen from the bulls of low fertilizing potential and hence, it becomes important to predict the fertility status before performing AI. In this study, the global proteomic profile of high fertile (HF) and low fertile (LF) buffalo bull spermatozoa was established using a high-throughput LC-MS/MS technique. A total of 1,385 proteins (≥1 high-quality PSM/s, ≥1 unique peptides, p < 0.05, FDR < 0.01) were identified out of which, 1,002 were common between both the HF and LF groups while 288 and 95 proteins were unique to HF and LF groups respectively. We observed 211 and 342 proteins were significantly high (log Fc ≥ 2) and low abundant (log Fc ≤ 0.5) in HF spermatozoa (p < 0.05). Gene ontology analysis revealed that the fertility associated high abundant proteins in HF were involved in spermatogenesis, sperm motility, acrosome integrity, zona pellucida binding and other associated sperm functions. Besides this, the low abundant proteins in HF were involved in glycolysis, fatty acid degradation and inflammation. Furthermore, fertility related differentially abundant proteins (DAPs) on sperm viz., AKAP3, Sp17, and DLD were validated through Western blotting and immunocytochemistry which was in coherence with the LC-MS/MS data. The DAPs identified in this study may be used as potential protein candidates for predicting fertility in buffaloes. Our findings provide an opportunity in mitigating the economic losses that farmers incur due to male infertility.

Nasal immunization with AMH-INH-RFRP DNA vaccine for improving follicle development and fertility in buffaloes.

Inhibin DNA vaccine has already been proven to improve the fertility of animals. This study aimed to investigate the effects of a novel Anti-Müllerian hormone (AMH)-Inhibin (INH)-RF-amide-related peptides (RFRP) DNA vaccine on immune response and reproductive performance in buffalo. A total of 84 buffaloes were randomly divided into four groups and nasally immunized twice a day with 10 ml of either AMH-INH-RFRP DNA vaccines (3 × 1010 CFU/ml in group T1, 3 × 109 CFU/ml in group T2, and 3 × 108 CFU/ml in group T3) or PBS (as a control) for 3 days, respectively. All animals received a booster dose at an interval of 14 days. ELISA assay revealed that primary and booster immunization significantly increased the anti-AMH, anti-INH, and anti-RFRP antibody titers in the T2 group compared with that in the T3 group. After the primary immunization, the antibody positive rate was significantly higher in the T2 group than that in the T3 group. In addition, ELISA results indicated that concentrations of E2, IFN-γ, and IL-4 were significantly higher in the antibody-positive (P) group compared to the antibody-negative (N) group. In contrast, there was no significant difference in the concentrations of P4 between the P and N groups. Ultrasonography results revealed a highly significant increase of 2.02 mm in the diameter of ovulatory follicles in the P group compared to the N group. In parallel, growth speed of dominant follicles was significantly higher in the P group than that in the N group (1.33 ± 1.30 vs 1.13 ± 0.12). Furthermore, compared to N group, the rates of oestrus, ovulation, and conception were also significantly higher in the P group. The novel AMH-INH-RFRP DNA vaccine improves the proportion of oestrus, ovulation, and conception in buffalo by promoting the production of E2 and the growth of follicles.

Association of Melatonin receptor 1A (MTNR1A) gene polymorphism with seasonal suppression of fertility in buffaloes

The polymorphism in Melatonin receptor 1A (MTNR1A) gene and its association with reproductive cyclicity (cyclic, n=51; non-cyclic, n=50) during summer season and age at first calving (&lt;3.5 years, n=38; &gt;3.5 years, n=60) was investigated in buffaloes subjected to blood sampling. Polymerase chain reaction (PCR) of genomic DNA was carried out using three primers viz. 824 bp (primer pair 1), 856 bp (primer pair 2) and 268 bp (primer pair 3) corresponding to exon-II of MTNR1A gene. Restriction fragment length polymorphism was performed using PCR products of primer pair 1 and 2. The digestion of PCR product of primer pair 1 with restriction enzymes viz. MnII, RsaI or Eco3II generated total restriction sites respectively as 14, 4 and 5. Based upon differential restriction sites, six genotypes revealed in buffaloes using MnII were MM-1 to MM-6, four genotypes using RsaI were RR-1 to RR-4, and four genotypes using Eco3II were EE-1 to EE-4. The digestion of PCR product of primer pair 2 with MnII, RsaI or Eco3II generated restriction sites respectively as 13, 5 and 6. Based upon differential restriction sites, six genotypes revealed using MnII were mm-1 to mm-6, five genotypes using RsaI were rr-1to rr-5, and three genotypes using Eco3II were ee-1 to ee-3. For primer pair 3, single stranded conformational polymorphism analysis exhibited 5 genotypes viz. AA, BB, AB, CC and DD. The results revealed that the buffaloes during summer had polymorphic genotypes for MTNR1A gene as buffaloes showing reproductive cyclicity had MM-3, MM-4, RR-2, EE-3, mm-2, mm-4, rr-3, ee-3, CC and DD genotypes, whereas their non-cyclic counterparts had MM-2, MM-6, EE-4, mm-3, mm-6, AA, BB and AB genotypes. However, no such differences were recorded with respect to age at first calving. In conclusion, polymorphic genotypes of MTNR1A gene may be considered as genetic markers to identify buffaloes with better reproductive potential during summer season.

Reproduction and Fertility of Buffaloes in Nepal.

Water buffalo (Bubalus bubalis) in Nepal contributes 57% of the total milk and 36% of the total meat production in the country. The productive efficiency of Nepalese buffaloes is quite low, due mainly to subfertility and infertility. Delayed puberty and prolonged inter-calving intervals, attributed mainly by anestrus due to silent cyclicity and ovarian acyclicity, are the major forms of infertility in Nepalese buffaloes. Moreover, buffaloes in Nepal show a distinct seasonal breeding pattern, with July to December as the active breeding season, and with April to June and January to March as the low and transitional breeding seasons, respectively. Endoparasitic infection and poor nutritional status, which are more severe during the low season, are found to be the major factors causing anestrus and compromising its treatment response in buffaloes. Various hormonal protocols for timed artificial insemination (TAI) have been attempted, with a varying pregnancy outcome. Recently, an integrated technique including anthelmintic treatment, nutritional supplementation and hormone-based fertility management programs for TAI has been developed and implemented successfully. A wider adoption of this technique as a package of practices could be key to improving the reproductive efficiency of buffaloes in Nepal.

Nano-depletion of morbid spermatozoa up-regulate Ca2+ channel, depolarization of membrane potential and fertility in buffalo

Despite recent advances in technique of spermatozoa cryopreservation, there are still ejaculates present that fail to meet strict quality standard; mainly due to detrimental effect of imbalance of free radicals. The omnipresence of dead/defective spermatozoa in ejaculates of eutherian species is a major source of excessive free radicals. Though sperm-selection techniques, as well as addition of antioxidants addressed the problem to a certain extent, the major source of free radicals in the semen remained, causing much damage. This study attempts to remove dead/damaged spermatozoa using negative fertility-marker. The effect is unraveled by Hypo-osmotic (HOS), and fluorescein-conjugated Pisum sativum agglutinin (FITC-PSA) assay, further confirmed by Ca2+-regulating mechanisms and depolarization of sperm membrane potential, reduction in concentration of free radicals and finally by in vitro fertility assay. The study involved functionalization of iron oxide nanoparticles (IONPs) with silane followed by bio-conjugation with anti-ubiquitin antibodies. The nano-purification of semen using anti-ubiquitin conjugated iron oxide nanoparticles (IONPs) (antibody concentrations 0.5, 1.0 and 2.0 μg/ml) was attempted. The efficiency of nano-purification was 18.1%–43.8% in the study. The results revealed greater (P ≤ 0.05) spermatozoa population with intact plasma membrane, acrosome integrity, high mitochondrial membrane potential and pattern-F (least intracellular Ca2+), evidence of low lipid peroxidation and higher total antioxidant capacity in nano-purified groups. More number of spermatozoa were bound to zona pellucida of matured oocytes from nano-depleted than non-depleted group. The findings demonstrate antibody concentration of 1.0 μg/ml bio-conjugated with IONPs as most efficient in enriching the ejaculate with functional spermatozoa with the highest percentage of zona binding.

Effect of Different Ecbolic Agents on Postpartum Reproductive Performance in Surti Buffaloes

The study was conducted to evaluate the effect of different ecbolic therapies on puerperium in twenty four Surti buffaloes divided into four different groups consisting of six animals in each. Buffaloes in Group-I (T1) were not given any treatment and considered as control group. Group-II (T2) and Group- III (T3) buffaloes were injected methylergometrine (5 mg) and dinoprost tromethamine (25 mg) i/m, respectively, immediately after parturition. The animals in Group-IV (T4) received oral herbal ecbolic, 100 mL, daily for first 10 days postpartum. The time required for placental expulsion, persistence of lochial discharge, involution of uterus and service period were significantly (P&lt;0.05) longer in T1 (14.86±6.65 h, 12.17±1.94 days, 36.50±4.76 days and 103.33±12.10 days, respectively) than T2 (10.33±2.71 h, 8.83±1.47 days, 33.33±3.61 days and 95.00±8.74 days), T3 (7.00±1.48 h, 8.00±1.41 days, 29.50±1.14 days and 86.50±4.88 days) and T4 (9.38±1.62 h, 10.17±1.47 days, 31.17±3.19 days and 97.80±7.21 days, respectively). The conception rates achieved in T1, T2, T3 and T4 groups/treatment were 50.00, 83.33, 100.0 and 83.33 %, respectively, with an overall conception rate of 79.17 % for all four groups of Surti buffaloes. The results were significantly superior with injection methylergometrine (5 mg) Group III given immediate postpartum, hence may be practiced to improve uterine health and postpartum fertility in buffaloes.

Effect of platelet-rich plasma (PRP) on post-thaw quality, kinematics and in vivo fertility of fertile and subfertile buffalo (Bubalus bubalis) spermatozoa.

This study investigated the effect of adding platelet-rich plasma (PRP) in semen extender prior cryopreservation on post-thaw quality, kinematics, and in vivo fertility of fertile and subfertile buffalo spermatozoa. Eleven buffalo bulls were classified based on their conception rate (CR) into fertile (n = 8, CR > 55%) and subfertile (n = 3, CR < 35%) groups. Ejaculates were collected with artificial vagina, pooled, and dispensed into 6 aliquots, diluted with Tris-egg yolk-glycerol extender supplemented with different proportions of PRP [0% (control), 5%, 10%, 15%, 20%, and 25%] followed by cryopreservation using standard procedures. Post-thaw sperm quality, kinematics, antioxidant activity, cryosurvival rate, and in vivo fertility were compared between fertile and subfertile groups and among proportions of PRP within each group. The results showed that 15% PRP greatly (P < 0.001) improved sperm characteristics, average path velocity, and curvilinear velocity of the subfertile group. Interestingly, 5%, 10%, and 15% PRP greatly (P < 0.001) reduced malondialdehyde content and improved enzymatic (glutathione peroxidase and superoxide dismutase) and total antioxidant capacity in fertile and subfertile groups. However, these three proportions of PRP significantly (P < 0.001) improved the cryosurvival rate of the subfertile group; only 15% PRP greatly improved CR of subfertile (60.83% vs. 34.17%) animals to be comparable with that of fertile ones treated with 5 (59.17%) and 10% (60.83%) PRP. In conclusion, adding 15% PRP to semen extender before cryopreservation is recommended to improve post-thaw quality, antioxidant activity, and in vivo fertility of buffalo semen particularly of the subfertile animals.

Effect of Milk Type Subjected to Different Heat Treatments on Cryo-Survivability and In Vivo Fertility of Buffalo (Bubalus bubalis) Spermatozoa in a Milk-Based Extender.

Oxidative stress is a major contributory factor to cellular damage during semen cryopreservation and results in a decreased fertilizing capacity of cryopreserved bull sperm. The inclusion of exogenous antioxidants sometimes exerts deleterious effects on sperm quality. Thus, enhancing the endogenous production of antioxidants is a requirement. This study aimed to investigate the effect of milk type heated at different temperatures on the antioxidant potential of extenders, and the subsequent post-thaw quality parameters and in vivo fertility of buffalo bull semen. Cow (C) and buffalo whole milk (B) were used separately for semen extender preparation, heated at five different temperatures (T1 = 90°C, T2 = 100°C, T3 = 110°C, T4 = 120°C, T5 = 130°C) for 10 minutes. Reactive sulfhydryl groups were measured in each subgroup by Ellman's reagents as CT1 = 143.2 μM, CT2 = 147.4 μM, CT3 = 151.5 μM, CT4 = 157.2 μM, CT5 = 161.8 μM, BT1 = 168.3 μM, BT2 = 172.5 μM, BT3 = 176.7 μM, BT4 = 196.3 μM, and BT5 = 205.7 μM. All semen samples were cryopreserved in milk-based extenders by using standard procedures. Post-thaw quality parameters including total and progressive motility, mitochondrial membrane potential, plasma membrane integrity, and acrosome integrity were found to be higher (p < 0.05) in the group (BT3) containing buffalo milk heated at 110°C, whereas in the same group, lipid peroxidation was found to be lower (p < 0.05) as compared with other treatment groups and control group. In vivo fertility of cryopreserved buffalo sperm was compared among BT3, CT1 (conventionally used milk extender), and a Tris egg yolk extender group. The fertility rates [47% (54/114), 30% (33/108), and 36% (37/103)] were higher (p < 0.05) in BT3 as compared with other groups. This study suggests that buffalo milk heated at 110°C has high antioxidant potential and improves post-thaw quality and in vivo fertility of cryopreserved buffalo bull semen.

Sexual Behaviour and Seminal Attributes Associated with Fertility of Jaffarabadi Buffalo Bulls

Background: The selection of breeding bulls based on the association of sexual behaviour, seminal attributes and conception rate are more important and economical. However, information on association of sexual behaviour and seminal attributes with fertility in buffaloes particularly with reference to Jaffarabadi buffalo bulls is meager. Methods: The study was carried out to assess the association of sexual behaviour and seminal attributes with first service conception rate (FCR) of Jaffarabadi buffalo bulls. A total of 192 semen ejaculates were evaluated and cryo-preserved. First AI conception rate (FCR) was carried out on 800 Jaffarabadi female buffaloes inseminated with frozen-thawed semen and pregnancy was confirmed by per rectal palpation at 2 months post-insemination. Result: The results indicated that both sexual behaviour and seminal attributes were associated with fertility in Jaffarabadi buffalo bulls.

Presynch-Heatsynch: A New Approach to Improve Ovarian and Fertility Responses in Water Buffalo (Bubalus bubalis)

Background: Standard estrus synchronization regimens resulted in variable outcomes in buffaloes. The present study evaluated ovarian and fertility responses following presynchronization in Heatsynch protocol administered in postpartum buffaloes. Methods: In group-I (Presynch-Heatsynch group, n = 30), PGF2α was administered on days -14 and -2. Then, GnRH analogue, PGF2α (Cloprostenol) and Estradiol benzoate were administered on day 0, 7 and 8, respectively followed by fixed-time artificial insemination (FTAI) 48 hours later. In group-II (Heatsynch group, n = 30), rest protocol was same, except that first two PGF2α injections were not administered. Result: The progesterone concentrations differed (P less than 0.01) between the two groups on days -2, 0 and 7. Post-treatment, progesterone profiles were also higher in pregnant compared to non-pregnant buffaloes in both the groups except on day 10. CL diameter differed (P less than 0.01) between groups on days -2, 0 and 7. It was larger in pregnant than non-pregnant buffaloes on day 7 in both the groups. Dominant follicle diameter remained larger on days -2, 0 and 8 in group-I than -II. Buffaloes getting pregnant had a larger (P less than 0.01) dominant follicle size on the day of FTAI in group-I than of group-II. Ovulatory response of 93.33 and 90.00% was observed in group-I and II. The conception rate was higher (66.66 vs. 40.00%; P less than 0.05) in group-I than Group-II. Presynchronization improved reproductive efficiency in Heatsynch treatment and may aid for better fertility in buffaloes.

Comparative biochemical profiles, utero-ovarian function, and fertility of the postpartum buffalo with and without subclinical endometritis.

In postpartum buffaloes, the process of uterine involution and changes in blood metabolic profile has not been studied in relation to development of subclinical endometritis (SCE). In this study, buffaloes (n = 100) approaching calving were identified. Weekly blood samples were collected on the day of calving up to 6weeks post-calving. The diameter of uterine horns and onset of ovarian cyclicity (corpus luteum) were recorded through ultrasonography. On the basis of polymorphonuclear cell (PMN) cell count in endometrial cytology at days 45-50 postpartum, buffaloes were divided into two groups, viz., with SCE (> 5% PMN; n = 38) and without SCE (≤ 5% PMN; n = 62). Buffaloes with SCE took longer (P < 0.05) time to complete uterine involution and had larger (P < 0.05) uterine horn diameter between 3rd and 6th weeks postpartum and lower prostaglandin F2α metabolite (PGFM) concentration on the day of calving (P < 0.05) and 1week (P < 0.001) post-calving than without SCE group. Buffaloes with SCE had lower (P < 0.001) concentration of glucose at weeks 2 and 3, higher (P < 0.001) β-hydroxybutyric acid (BHBA) at week 3, and lower serum albumin concentration throughout the sampling period (P < 0.05 to 0.001) except at 1week post-calving as compared to without SCE group. The urea concentration was significantly lower (P < 0.05 to 0.001) in buffaloes with SCE from 4weeks post-calving onwards than without SCE group. The calcium concentration was lower in buffaloes with SCE at weeks 5 (P < 0.001) and 6 (P < 0.05) postpartum, whereas the concentration of magnesium and phosphorus was uniform between the two groups. No significant (P > 0.05) difference in onset of ovarian cyclicity between the 2 groups was observed, whereas buffaloes with SCE had longer (P = 0.001) median days open (141days) than their counterpart (117days). The first service conception rate, cumulative pregnancy rate, and pregnancy rate at 150days postpartum were lower (P < 0.05) in buffaloes with SCE than without SCE group. In summary, higher BHBA and lower serum concentrations of glucose, albumin, urea, and calcium control onset of subclinical endometritis which in turn has negative impact on fertility of buffaloes.

Effect of Three Diluent Types and Equilibration Times on the Quality and Fertility of Buffalo (Bubalus bubalis) Semen

Effect of Three Diluent Types and Equilibration Times on the Quality and Fertility of Buffalo (Bubalus bubalis) Semen

On the Effect of the Temperature-Humidity Index on Buffalo Bulk Milk Composition and Coagulation Traits.

Little is known about the effects of high levels of environmental temperature and humidity on milk yield and quality in buffaloes since this species is known to be more heat tolerant than cattle. However, the distribution of sweat glands and the dark skin color can negatively affect heat tolerance. Moreover, due to increased global temperatures, concerns regarding heat stress and thermoregulation in dairy animals, including buffaloes, have been extended to the northern hemisphere. In this study, the effects of both the temperature-humidity index (THI) and the maximum daily temperature-humidity index (MTHI) were estimated on bulk milk traits, namely fat, protein, lactose, urea content, pH levels, somatic cell score, coagulation properties, and bacteria count. The dataset consisted of repeated data from 99 Mediterranean water buffalo farms, and mixed models were used for the analyses. Supporting the negative correlations observed, bulk milk fat, protein, and lactose content were significantly lower when THI and MTHI were higher. Similarly, milk pH was lower when THI and MTHI were high; however, high levels of THI or MTHI seemed to not be markedly associated with the milk's coagulation ability. According to both analysis of variance and correlations, the somatic cell score was not significantly affected by the THI and MTHI. This is the first study based on a large dataset that evaluates the impact of high temperature and humidity in Italian buffalo milk and that provides correlations with traits of interest for the dairy industry, i.e., milk acidity and coagulation ability. In general, findings show that the effects of elevated THI and heat stress on bulk milk quality in buffalo is less evident than in cattle. These preliminary results intend to open debate on the issue of heat stress in dairy buffaloes that are reared in temperate regions. Further studies should focus on individual milk and performance and should investigate the relationship between high THI and buffalo fertility, behavior, and welfare.

Seasonal effects on miRNA and transcriptomic profile of oocytes and follicular cells in buffalo (Bubalus bubalis)

Season clearly influences oocyte competence in buffalo (Bubalus bubalis); however, changes in the oocyte molecular status in relation to season are poorly understood. This study characterizes the microRNA (miRNA) and transcriptomic profiles of oocytes (OOs) and corresponding follicular cells (FCs) from buffalo ovaries collected in the breeding (BS) and non-breeding (NBS) seasons. In the BS, cleavage and blastocyst rates are significantly higher compared to NBS. Thirteen miRNAs and two mRNAs showed differential expression (DE) in FCs between BS and NBS. DE-miRNAs target gene analysis uncovered pathways associated with transforming growth factor β (TGFβ) and circadian clock photoperiod. Oocytes cluster in function of season for their miRNA content, showing 13 DE-miRNAs between BS and NBS. Between the two seasons, 22 differentially expressed genes were also observed. Gene Ontology (GO) analysis of miRNA target genes and differentially expressed genes (DEGs) in OOs highlights pathways related to triglyceride and sterol biosynthesis and storage. Co-expression analysis of miRNAs and mRNAs revealed a positive correlation between miR-296-3p and genes related to metabolism and hormone regulation. In conclusion, season significantly affects female fertility in buffalo and impacts on oocyte transcriptomic of genes related to folliculogenesis and acquisition of oocyte competence.

Different times to perform timed artificial insemination when using a P4/E2/eCG-based protocol in buffalo

ABSTRACT: The aim of this study was to evaluate different times for timed artificial insemination (TAI) in buffalo submitted to a P4/E2/eCG-based protocol. In this study, 204 buffaloes were distributed into one of two groups (TAI56, n=103 and TAI64, n=101). At a random stage of the oestrous cycle (Day 0 = D0), in the morning (TAI56, a.m.) or afternoon (TAI64, p.m.), buffaloes received an intravaginal progesterone device (P4; 1.0 g) plus EB (2.0 mg i.m.). On D9 a.m. (TAI56) or p.m. (TAI64), the P4 was removed and buffaloes received PGF2a (0.53 mg i.m. sodium cloprostenol) and eCG (400 IU i.m.). On D10 a.m. (TAI56) or p.m. (TAI64), 24 h after P4 removal, buffaloes were treated with EB (1.0 mg i.m.). Buffaloes from TAI56 and TAI64 were inseminated 56 and 64 h after P4 removal (D11, p.m. and D12, a.m., respectively). Ultrasound examinations were performed on D0 to ascertain ovarian follicular status, at TAI to measure the diameter of the dominant follicle (DF) and D42 for pregnancy diagnosis. The statistical analysis was performed using the GLIMMIX procedure of SAS®. There was no difference between TAI56 and TAI64 for the diameter of the DF at TAI and the pregnancy per TAI. It was concluded that TAI 56 or 64 h after P4 removal did not affect fertility in buffaloes submitted to the induction of ovulation with EB. The present research supports that is possible to perform TAI at any time throughout the day in buffalo synchronized during the non-breeding season.