The study aimed to describe the Bubalus bubalis seminal plasma proteome using a label-free shotgun UDMS approach. A total of 859 nonredundant proteins were identified across five biological replicates with stringent identification. Proteins specifically related to sperm maturation and protection, capacitation, fertilization and metabolic activity were detected in the buffalo seminal fluid. In conclusion, we provide a comprehensive proteomic profile of buffalo seminal plasma, which establishes a foundation for further studies designed to understand regulation of sperm function and discovery of novel biomarkers for fertility. MS data are available in the ProteomeXchange with identifier PXD003728.

The aim of the present review is to provide complementary information about reproductive characteristics of male buffaloes having as perspective a four -year-study based on a weekly seminal collection schedule and collection of data in partner properties of several regions of Brazil. Aspects of testicular growth, reproductive behavior and characteristics of 1477 ejaculates of 13 donors are discussed in conjunction with data available in the literature.

The objective of this study was to evaluate the fertility of buffalo semen for in vitro embryo production (IVEP) by comparing the effectiveness of refrigerated versus frozen semen. Three OPU sessions were held at 30-day intervals. For oocyte fertilization three buffalo bulls were used, one per session. At each OPU-IVEP session, one ejaculate was collected and divided into two equal aliquots. Each aliquot was either refrigerated at 5ºC/24 hours or frozen. A TRIS extender containing 10% low density lipoproteins, 0.5% lecithin and 10 mM acetylcysteine was used adding 7% glycerol for freezing. Sperm motility/kinetic was evaluated by CASA and sperm membrane integrity by the hypoosmotic swelling test. The evaluations were performed at 0 h (post final dilution at 37ºC), at 4 and 24 hs post-incubation at 5ºC and post-thaw. At 24 hs incubation and immediately post thaw sperm cells were used for in vitro fertilization of buffalo oocytes equally distributed between both groups. Cleavage rates and embryo development were followed. The embryo/matured and embryo/cultured rates were 25.4 x 14.0% and 29.4 x 18.5% (P<0.05), for chilled and frozen semen, respectively. It is concluded that cooled semen can be used for in vitro embryo production in buffalo and that a better efficiency may be expected for cooled compared to frozen semen.

O sêmen de 11 touros búfalos, da raça Murrah, com idade variando de cinco a seis anos foi coletado por vagina artificial e submetidos às análises das características físicas e morfológicas. O experimento objetivou testar in vitro a eficácia de quatro diferentes extensores: Controle (TRIS com 10% LDL) e o mesmo extensor nas concentrações de 0,5; 1,0 e 2,0% de lecitina de soja sobre a longevidade espermática de búfalos no processo de refrigeração a 5 oC por 1, 24, 48, 72, 96 e 120 horas. Após a coleta cada ejaculado foi fracionado em 4 alíquotas, para a obtenção de 50x106 SPTZ/mL, envasado em palhetas de 0,5 mL, refrigerado durante 4 horas obtendo-se uma curva de resfriamento de 0,25 oC/minuto. As palhetas foram mantidas refrigeradas a 5 oC e sequencialmente avaliadas, reaquecendo-se o conteúdo a 37 oC/30 segundos antes da avaliação da motilidade espermática (total e progressiva) utilizando o sistema computadorizado (CASA). Foram retiradas amostras para avaliação pelo teste hiposmótico (HOST) nos tempos 1, 24, 48 e 72 horas. Para a análise estatística utilizou-se o pacote STATA 12.0, sendo os valores obtidos submetidos ao Teste de Friedman (P < 0,05). Os espermatozoides progressivos e a integridade da membrana plasmática foram superiores (P < 0,05) no extensor de TRIS com 10% LDL acrescido de 0,5% Lecitina de soja (67,2 e 67,9%) no processo de refrigeração por 48 horas. Em conclusão, avaliação comparativa dos parâmetros espermáticos obtidos revelou que o extensor TRIS com 10% LDL acrescido de 0,5% de lecitina de soja, pode ser efetivamente usado para a preservação do sêmen de búfalo refrigerado em IA e IATF por até 48 horas.

The objective of this study was to compare the reproductive efficiency of dairy buffaloes undergoing fixed-time artificial insemination (FTAI) protocols based on progesterone/estrogen (P4/E2) and eCG during unfavorable breeding season using cooled (CS) and frozen semen (FS). A total of 446 buffaloes (> 40 days postpartum) were randomly distributed into four blocks (years): B1-2014 (n = 143), B2-2015 (n = 34), B3-2016 (n = 90), and B4-2017 (n = 179). Each block was subdivided into two (AI with CS and FS using the same ejaculate of each bull). Thus, the block subdivision was as follows: B1 (CS = 71 and FS = 72); B2 (CS = 18 and FS = 16); B3 (CS = 47 and FS = 43); and B4 (CS = 90 and FS = 89). The ejaculates of eight Murrah bulls collected using an artificial vagina were divided into two aliquots: one aliquot was diluted in Botu-Bov® commercial extender and cooled (BB-CS), and the other was diluted in the same extender and frozen (BB-FS). BB-CS aliquots were cooled at 5 °C/24 h using a refrigerator. BB-FS group aliquots were also cooled, and after equilibrating at 5 °C for 4 h, were placed in a 21-L Styrofoam box, 5 cm above the surface of liquid nitrogen. In the afternoon (A) on D0 (2:00 p.m.) the animals received EB 2.0 mg IM (Estrogin®) and an ear implant (CRESTAR® 3.0 mg P4). At D9 (A), the implant was removed, and the animals received eCG 400 IU IM (Folligon® 5000) + Cloprostenol PGF2α 0.530 mg IM (Sincrocio®). At D10 (A), the animals received EB 1.0 mg IM (Estrogin®), and at D12 (8:00 a.m.), AI was performed. At D42, pregnancy was diagnosed via ultrasonography. Total CRs were 48.2% CS and 34.6% FS for years 2014 to 2017, with a significant difference of 13.7% (P<0.05). In conclusion, cooled semen resulted in higher CR than frozen semen in dairy buffaloes under the P4/E2 and eCG FTAI during the unfavorable reproductive season.