This study aimed to verify how seasonal hydrology affects artificial insemination strategies in northern Brazil to promote better livestock management and ecosystem sustainability. A total of 1780 inseminations were analyzed, including 1590 fixed-time artificial inseminations (FTAI) and 190 conventional artificial inseminations (CAI) (AI in natural estrus without using hormones) conducted over ten years. Of these, 370 inseminations occurred during the flood season (January to May), 700 during the intermediate season (June to September), and 710 during the ebb season (October to January). The estrus synchronization protocols were utilized according the ovarian cyclicity of buffaloes, i.e.: Protocol I (Ovsynch), was used for cycling buffaloes; Protocol II (Ovsynch-P4); Protocol III (CL-synch), for the buffaloes with a mature corpus luteum (CL); Protocol IV (P4 + BE) + (PGF2α + eCG) + GnRH and Protocol V (P4 + VE) + (PGF2α + eCG) + BE were implemented for buffaloes in anestrus, with low body condition, or during unfavorable seasons. AI was performed using cryopreserved-thawed semen. Pregnancy diagnosis was performed using ultrasonography or rectal palpation between 45 and 90 days after insemination. A higher pregnancy rate was observed for the PI (66.87%) than for II (53.77%), III (52.41%), IV (53.35%) and V (33.17%). Regarding seasons, the overall pregnancy rate (53.14%) for FTAI was significantly higher (P < 0.05) than CAI (27.4%). During the season of Full tide, Intermediary and Ebb tide, the percentage of pregnancy rate for FTAI vs CAI was 48.28 vs 20.00, 47.77 vs 10.00 and 61.47 vs 41.00, respectively. The pregnancy rate was significantly higher during ebb tide for both FTAI/CAI than in other seasons. In conclusion, seasonal hydrology impacts the outcome of AI strategies in buffaloes in the Amazon region of northern Brazil. A suitable synchronization protocol can enhance buffalo fertility and improve agricultural practices.

This study aimed to verify the effect of GnRH at the time of artificial insemination and Dominant follicle (DF) diameter on the Pregnancy rate (PR). 384 multiparous upgraded Murrah breed buffalo within 30 to 60 days postpartum with a body condition score of 3.4±0.6 were used. The synchronization protocol consisted of a day 0 (D0) intravaginal device of 1g of P4, application of 2.0mg of BE, and administration of 10 ml of injectable mineral supplementation at 4 pm. On D9, at 4 pm, the device was removed, and injected 0.265 mg of PGF2α and 300 IU of eCG Intramuscular (IM). On day 11, females were categorized based on the size of their dominant follicles. Females with ≥ 13 mm DF were divided into two groups: group 1 received GnRH, while group 2 did not. Similarly, females with < 13 mm DF were split into Group 3, which received GnRH, and Group 4, which did not. At 4 pm, 0.1 mg of GnRH was administered IM according to their groups. On D12, 16 hours after the injection of GnRH (at 8 am), Fixed-time artificial insemination (FTAI) was performed. The pregnancy was diagnosed after 30 days of FTAI by ultrasound. The overall PR was 53.9% (207/384). No significant difference was observed between the groups and follicular size. More pregnancies were observed when the follicle size was around 13mm. It was concluded the administration of GnRH could enhance the efficiency of FTAI in buffaloes.

This study aimed to investigate the effects of refrigeration and cryopreservation on the sperm kinetic and morpho-functional parameters of 5/8 Girolando bulls. Eleven ejaculates from each bull were collected, diluted, and divided into two portions: one portion was subjected to refrigeration at 5ºC for 24 and 48 hours, while the second portion underwent cryopreservation(-196ºC). Post-thawing, various kinetic parameters were analyzed using the Computer Assisted Sperm Analysis (CASA) system. These parameters included total motility (TM), progressive motility (PM), curvilinear velocity (VCL), straight line velocity (VSL), average path velocity (VAP), linearity (LIN), straightness coefficient (STR), wobble coefficient (WOB), amplitude of lateral head displacement (ALH) and beat cross frequency (BCF). Additionally, plasma membrane integrity (PMI), acrosomal membrane integrity (AMI), mitochondrial membrane potential (MMP), and chromatin condensation (CC) were examined for morpho-functional analyses. The results indicated that kinetic parameters (TM, PM, VCL, VSL, VAP) and morpho-functional analyses (PMI, MMP) differed significantly (P < 0.05) between cryopreserved and refrigerated semen, as well as other parameters such as LIN, ALH, and BCF across all groups. The acrosomal membrane integrity showed no differences, but CC varied significantly among all groups. In conclusion, semen from 5/8 Girolando bulls can be preserved for up to 48 hours at 5ºC, providing an alternative for the short-term use of genetic material from this breed.

Mathematical modeling of the estrous cycle could help in understanding the dynamics of this complex biological system. Thus, this study aimed to use a mathematical model that accounts for the variations of progesterone during the estrous cycle in Canindé goats. Five adult (3-6 years old) and six young (1-2 years old) healthy and cyclic goats were used. The females were submitted to a hormonal estrus synchronization treatment and blood samples were collected for progesterone dosage from the day of synchronized estrus until the next natural estrus. Peripheral plasma progesterone levels were compared between age groups. The progesterone curves were fitted to mathematical models for both luteal and luteolytic phase. In conclusion, the proposed model, which is simple, makes it possible to characterize the evolution of the progesterone level during the estrous cycle of Canindé goats by few parameters. It is then possible to use these parameters to analyze other effects.

For approximately 40 years, microencapsulation technology has been utilised across various species due to its ability to release semen gradually after artificial insemination. This study aimed to establish the use of the alginate microencapsulation procedure for goat semen and to investigate whether this method enhances longevity during cold storage compared to the traditional straw method. Semen was collected from Canindé bucks and analysed using Computer-Assisted Semen Analysis (CASA). The semen was then diluted in a commercial extender and packaged either in straws or microcapsules (using 1% sodium alginate). Both groups were refrigerated at 4°C-5°C and assessed at 24, 48 and 72 h after dilution. The evaluation included assessments of sperm viability, abnormalities, membrane integrity, and DNA integrity. Data were analysed using repeated-measures ANOVA at p < 0.05. Concerning the parameters straight line (VSL) and average path (VAP), no statistical differences (p > 0.05) were observed. However, the microcapsule group showed significantly higher results (p < 0.05) for straightness (STR), beat cross frequency (BCF), and wobble (WOB) at 24 and 48 h of storage. Sperm viability was also higher (p < 0.05) in the microcapsule group at 24 and 48 h of storage. In conclusion, this study demonstrates the feasibility of microencapsulating goat semen. Further in vivo and/or in vitro fertility trials are needed to confirm these findings.

The present study was conducted to evaluate the conception rate after fixed time natural mating and artificial insemination after synchronization of estrus in murrah buffaloes. Total 16 murrah buffaloes were selected and randomly divided into two groups, one for natural mating and one for artificial insemination. Buffaloes received estrus synchronization protocol that is, 2mg Oestradiol benzoate intra muscular + 1.38gm Progesterone intra-vaginal on day 0, 12.5mg Dinoprost + 400 IU PMSG intra muscular + removal of the progesterone insert on day 9, 0.0105mg Buserelin acetate intra muscular on day 11 and followed by fixed time natural mating and artificial insemination on day 12 and 13 according to the group. The status of pregnancy was examined by rectal palpation on day 90 post insemination (natural and artificial). All the animals were expressed heat signs and the conception rate was 8/8 (100%) and 5/8 (62.5%) in natural mating and artificial insemination group, respectively. In conclusion, fixed time natural mating may be helpful for the farmers having a small herd of buffaloes.

The reproductive management of the buffalo species still faces several unresolved problems, which directly affect the productivity of the herd, one of them being the presence of repeat breeder females. Given this scenario, this study aimed to verify the developmental competence of oocytes obtained from repeat breeder females and submitted to parthenogenetic activation. In addition, embryo gene expression was compared to normally fertile females. Murrah buffaloes were divided into two groups: repeat breeder (RB, n = 8) and normally fertile or control (CR, n = 7). Cumulus-oocyte complexes (COCs) were aspirated by transvaginal ovum pick-up from estrus synchronized females. The COCs were submitted to IVM for 24 h, and subsequently, the oocytes were activated using ionomycin, followed by 6-DMAP. Afterwards, the presumptive parthenotes were cultured for six or seven days in a microenvironment of 5 % CO2, 5 % O2, and 90 % N2 at 38.5 °C. The expression of OCT4, GLUT1, BCL2 and TFAM genes from blastocysts was evaluated. The overall COCs recovery rate was 70.9 % (190/268). The maturation (57.8 vs 71.1), cleavage (45.2 vs 62.2) and blastocyst (30.1 vs 45.9) rates did not differ (P > 0.05) between RB and CR females, respectively. Similarly, no significant difference (P > 0.05) was observed for the expression of studied genes in both RB and CR females. In conclusion, oocytes obtained from RB were as developmentally competent as those collected from CR females, with similar energy metabolism and in vitro development capacity. Thus, the low fertility rate of repeat breeder buffaloes, when compared to normal cyclic females, must be due to subsequent events to the blastocyst stage.

This study assessed the histones methylation profile (H3K4me3 and H3K9me3) in late preantral (PA) and early antral (EA) caprine follicles grown in vivo and in vitro, and the anethole effect during in vitro culture of PA follicles. Uncultured in vivo-grown follicles (PA, n = 64; EA, n = 73) were used as controls to assess the methylation profile and genes' expression related to apoptosis cascade (BAX, proapoptotic; BCL2, antiapoptotic), steroidogenesis (CYP17, CYP19A1), and demethylation (KDM1AX1, KDM1AX2, KDM3A). The isolated PA follicles (n = 174) were cultured in vitro for 6 days in α-MEM+ in either absence (control) or presence of anethole. After culture, EA follicles were evaluated for methylation, mRNA abundance, and morphometry. Follicle diameter increased after culture, regardless of treatment. The methylation profile and the mRNA abundance were similar between in vivo-grown PA and EA follicles. Anethole treatment led to higher H3K4me3 fluorescence intensity in EA follicles. The mRNA abundances of BAX, CYP17, and CYP19A1 were higher, and BCL2 and KDM3A were lower in in vitro-grown EA follicles than in vivo-grown follicles. In conclusion, in vitro follicle culture affected H3K4me3 fluorescence intensity, mRNA abundance of apoptotic genes, and steroidogenic and demethylase enzymes compared with in vivo-grown follicles.

Crotalicidin is a cathelicidin-related anti-infective (antimicrobial) peptide expressed in the venom glands of the South American rattlesnake Crotalus durissus terrificus. Congener peptides of crotalicidin, named vipericidins, are found in other pit vipers inhabiting South America. Crotalicidin is active against bacteria and pathogenic yeasts and has anti-proliferative activity for some cancer cells. The structural dissection of crotalicidin produced fragments (e.g., Ctn [15-34]) with multiple biological functionalities that mimic the native peptide. Another structural characteristic of crotalidicin and congeners is a unique repetitive stretch of amino acid sequences in tandem embedded in their primary structures. One of the encrypted vipericidn peptides (Ctn [1-9]) was synthesized, and the analog covalently conjugated with rhodamine B (RhoB-Ctn [1-9]) displayed considerable antimicrobial activity and selective cytotoxicity. Methods to evaluate antimicrobial peptides' toxicity include lysis of red blood cells (hemolysis) in vitro and cytotoxicity of healthy cultured cells (e.g., fibroblasts). Here, as a non-conventional model of toxicity, the bovine oocytes were exposed to two standardized concentrations of RhoB-Ctn [1-9], and embryo viability and development at its first stage of cleavage (division of cells) and blastocyst formation were evaluated. Oocytes treated with peptide at 10 and 40 μM induced cleavage rates of 44.94% and 51.53%, resulting in the formation of blastocysts of 7.07% and 11.73%, respectively. Light sheet microscopy and in silico prediction analysis indicated that RhoB-Ctn [1-9] peptide interacts with zona pellucida and internalizes into bovine oocytes and developing embryos. The ADMET prediction estimated good bioavailability of RhoB-Ctn [1-9]. In conclusion, the peptide appeared harmless to bovine oocytes and, remarkably, activated the parthenogenesis in vitro.

Clinical evaluations have shown that repeat breeding is a major cause of infertility in buffaloes. The follicular fluid (FF) composition reflects directly metabolic status and fertility in females. Given this scenario, this study aimed to perform quantitative proteomic analysis of the FF collected from normal cycling (NC) and compared to repeat-breeder females. According to farm records, buffaloes were divided into two groups: normal cycling (NC, n = 7) and repeat-breeder (RB, n = 8) females. After estrus synchronization and using ultrasound-guided ovum pick-up, FF was aspirated from large follicles (> 8 mm). Posteriorly, proteins were identified by the shotgun method. A total of 119 proteins were identified and, among theses, three were uncharacterized and a protein (LOC123334375) was identified only in the NC group. The protein HP-25 homolog 2 was expressed only in RB females. The LFQ (label-free quantitation)-intensity of the proteins afamin (AFM), transthyretin (TTR), clotting factor IX (F9) and Xaa-Pro dipeptidase (PEPD) was significantly (P < 0.05) higher in RB than in NC females. In conclusion, the use of quantitative proteomics proved to be an important tool for the study of RB in buffaloes. The identification of HP-25 homolog 2 protein only in the RB females suggests that it can be used as a biomarker for this reproductive disorder. Additionally, an uncharacterized (LOC123334375) protein was identified only in normal cycling females and compensatory oxidative stress proteins were more expressed in repeat-breeders.

Under natural and well-managed conditions, the buffalo has good reproductive and productive indices. However, in vitro embryo production (IVEP) has been used commercially to maximise the number of elite animals. In this species, several factors (donor management, in vitro culture medium, semen, in vitro conditions, embryo transfer) still affect the IVEP results. In addition, the cost of this technique is very high for this purpose. Therefore, more studies, as well as adequate plans, are needed to achieve this objective efficiently. In this review, we discussed the current commercial status, influencing factors (in vivo and in vitro), and the progress and future challenges of IVEP in buffalo. A total of 81 references were used from 1979 to 2022. The relevant data or literature were searched using the following databases: Google, ResearchGate, Science Alert, Science Direct and PubMed, using the following keywords: buffalo oocytes/COCs, buffalo embryos, pregnancy and calving or live birth rate after embryo transfer. The best maturation, cleavage and blastocyst rates in the in vitro production of buffalo embryos were 95.8, 75.2 and 33.4%, respectively. The pregnancy and live birth rates ranged from 22.2% to 43.5% and from 15.3% to 36.5%, respectively, after the transfer of fresh embryos produced in vitro to the recipients. This review will help to contextualise IVEP in buffaloes, as well as create an adequate plan for implementing IVEP in buffaloes.

A pluriparous Murrah buffalo was presented with vaginal tear over fornix vagina along with post-partum prolapse of uterus, cervix and urinary bladder. The case was managed by suturing of vaginal wall after repositioning the prolapsed mass. The catheterization of urinary bladder and modified Buhner's sutures through vulvar lips were applied to prevent the recurrence of the condition.

The present research work was conducted to investigate the incidence of subclinical endometritis (SE) in repeat breeder dairy cattle, and to study the comparative efficacy of different antimicrobial and polyherbal intrauterine therapies. Repeat breeding crossbred cows (>90 days in milking) with clear cervico-vaginal mucous discharge in spontaneous estrus were examined for uterine cytology by the cytobrush technique. The cows with ≥ 4% polymorphonuclear cells (PMNs) in uterine cytology were considered to be affected with SE. These cows were randomly and equally divided (n=12/group) into 5 groups and treated with the intrauterine medicine Vodine-IU (T1), Metricare-IU (T2), Utriguard-IU (T3), Uraksha (polyherbal preparation) (T4), or left untreated (control/C). In the subsequent estrus, uterine cytology was repeated and the cows were inseminated. The cows that failed to conceive but showed estrus were re-inseminated. The pregnancy status was checked. The incidence of SE was observed to be 36.80% (60/163). The PMNs percentage significantly declined in all the treatment group cows in response to treatment. The highest conception rate was observed in the T4 group (Uraksha treated) cows. It was concluded that the uterine cytobrush technique can be used efficiently for diagnosis of SE in dairy cattle, and polyherbal intrauterine drugs may be a good alternative in intrauterine antimicrobial therapy.

The present study was aimed to evaluate the efficiency of estradiol cypionate, estradiol benzoate and gonadotropin-releasing hormone during fixed-time artificial insemination (FTAI) protocols in multiparous Murrah buffaloes. A total of 35 buffaloes were used between September and November 2021 and subjected to a synchronization protocol. On day 0, insertion of an intravaginal progesterone device (CIDR, 1.38 g) was made with intramuscular (i/m) injection of estradiol benzoate (EB, 2 mg); On day 9: CIDR removed with i/m injections of Dinoprost (12.5 mg) and eCG (400 IU), and then buffaloes were randomized into three treatment group: 1) EB 1 mg on day 10 (n=13). 2) estradiol cypionate (EC) 1 mg on day 9 (n=12), and 3) Buserelin acetate (GnRH) 0.0105 mg on day 10 (n=10). The estrus signs observed during the FTAI (56 h after CIDR withdrawal) and conception rate at 90 days after FTAI were recorded. The conception rates were 61.5, 58.3 and 60.0% for EB, EC and GnRH treatment groups, respectively. There was no significant difference between the treatment groups for conception rate and signs of estrus. Therefore, it was concluded that all three treatments can be efficiently used during estrus synchronization protocol with FTAI in multiparous Murrah buffaloes.

Cryopreservation of oocytes and embryos is a supplementary technique for in vivo and in vitro embryo production. In the last decade, the technique has been improved but did not achieve a satisfactory commercial level. Also, the results were variable. The pregnancy and live birth rates were 16 to 52% and 5.3 to 26.9% after implantation of vitrified embryos, respectively (from the year 1993 to 2020). In final consideration, the quality of oocytes and embryos is the most important factor to obtain better results after cryopreservation. The addition of antioxidant and lipid-depleting agents to the medium is a factor that improves cryogenic tolerance. Additionally, two-step vitrification is a reliable method for cryopreservation. Studying epigenetic effects and introducing lipidomic, transcriptomic and proteomic technologies can help improve this technology.

Buffalo is a multipurpose and economically important animal due to the demand for its products (milk and meat). Thus, the use of reproductive biotechnologies is important to maximize the diffusion of genetically superior dams and sires. After the unsatisfactory results of the Multiple Ovulation and Embryo Transfer, the combined effect of ovum pick-up from live animals and in vitro embryo production (IVEP) has great potential to dissemination of selected genetics in buffalo herds, contributing to an increase in meat and milk production. During the past two decades, considerable advances have been made in IVEP following continuous scientific effort, but at the moment their cost is not satisfactory for commercial purpose. This technique is refined day by day in order to improve the buffalo embryo quality. Thus, the objective of this paper was to review the state-of-art in IVEP, as well as discussed the emerging technologies that can contribute to improving the results of this technology in buffalo species.

Two rare cases of Schistosomus Reflexus (SR) and Fetal Arthrogryposis (FA) in ewe were successfully managed by vaginal

In the last some years, veterinarians are now frequently requested to solve fertility problems in the dogs, mainly due to the increased popularity of purebred dogs as well as for sentimental or financial reasons. Endometritis is one of the very common post-partum/post estrus complication that usually remains unnoticed until the appearance of notifiable clinical sings. Primary sins observed by the clinician is purulent vaginal discharge, may be associated with fever, dehydration, anorexia. Reports have shown that E. coli is the main bacterium associated with it but some case also shows mixed infection of several bacterium. In the case of nursing mother care of puppies also challenging due to dam unable to nurse them or puppies have been isolated due to the antibiotic residue found in dam’s milk. History and clinical signs are the first approaches for diagnosis, it is more adventitious to diagnose by the use of modern technique like ultrasonography, ABST and blood picture profile. Antibiotics and ecbolic drugs with supportive therapy give results in most of the case if the patient in stable condition.