Utilização do choque osmótico na avaliação da viabilidade de sêmen criopreservado de ovinos

This study evaluated the use of hipoosmotic swelling test (HOST) with deionized water (0 mOsmol), as a method of post thaw ram semen evaluation and correlate their findings with different techniques of semen evaluation. Therefore, twenty semen samples of 20 different adult rams were assessed as for kinetic sperm parameters through computerized system (IVOS 12, Hamiton Thorn Biosciences, Beverly, MA, EUA) and subjective analysis. The sperm membranes viability was carried out by the association of fluorescent probes (propidium iodide, JC-1 and FITC-PSA). The structural integrity of the plasma membrane was also studied through supravital test with eosin and the functional integrity of membrane evaluated by doing the hipoosmotic swelling test with deionized water (0 mOsmol), in the following proportions: One part of semen for 10 (HOST 10), 50 (HOST 50) and 100 (HOST 100) parts of water. After semen dilution in the different proportions it was fixed in formalin-buffered saline and analyzed with regard to percentage of HOST reactive sperm (bent/coiled). The percentage of reaction obtained for HOST 10 (33,1%); HOST 50 (32,8%) and HOST 100 (31,8%) did not differ significantly. HOST 10 presented positive correlation with the plasma membrane integrity by the EOS (r = 0,80; p < 0,05). Positive correlations between HOST 50 and HOST 100 with sperm subpopulation with membrane integrity by fluorescence were observed (r = 0,83 and r = 0,85; p < 0,01). The findings suggest that the HOST with deionized water can provide additional information for post thawing ram sperm viability evaluation.

Influence of sperm motility factors on spermatozoa obtained from diffent epididymal segments

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Influência de aspectos técnicos e anatômicos nos índices de fertilidade baseado no desempenho da inseminação artificial

The aim of this work was to examine both the influence of anatomical and technical aspects on fertility rate of sheep based on the performance of transcervical artificial insemination (TCAI). Transcervical artificial insemination was performed with traction of the cervix in 122 ewes using frozen semen from 11 rams, both Santa Ines breed. The data collected were: type of external cervical opening (CO) (P - papilla; FL - flap; DB - duckbill, S - spiral; RO - rosette), duration of cervical manipulation (2-3, 4-5 and 6-7 minutes), degree of difficulty in cervical transposition (low, moderate, high) and presumed semen deposition site (SC - superficial cervical; DC - deep cervical; IU - intrauterine). The influence of these variables on pregnancy rate was evaluated. Cervical opening type and duration of cervical manipulation had no influence (p>0.05) on fertility. The degree of difficulty in cervical manipulation influenced (p<0.05) pregnancy rate, since insemination classified as low grade had 52% of pregnancy, while those classified as high recorded only 20%. The presumed site of semen deposition influenced significantly (p<0.05) fertility. Pregnancy rates of deposition at each site were: UI – 45.8%, DC – 25.7%; SC – 15.4%. As expected, deeper depositions resulted in higher fertility. In conclusion, the performance of TCAI did not depend on the anatomical classification of external cervical opening of ewe and the duration of cervical manipulation within the range tested (2-7 minutes). The TCAI may have higher fertility rates if difficulties in the application were reduced and the semen deposition was deeper.

Utilização de sistemas de refrigeração de sêmen equino na estabilização das amostras seminais previamente à congelação

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Retorno à atividade reprodutiva de touro angus após fimose traumática com obliteração parcial da luz prepucial: relato de caso

The main disturbs affecting the genital segment of bulls are balanitis, phimosis and preputial injuries, abscesses or prolapses, resulting in decreased libido or mating ability. One case of traumatic phimosis with partial obliteration of preputial lumen is reported in Aberdeen Angus bull. During anamnesis, it was reported that the animal was unable to expose the penis and presented increased penile and preputial volume of unknown etiology after being submitted to a libido test. During clinical examination, a firm preputial mass of 15 cm in diameter was observed in the middle third of the prepuce and stenosis of preputial lumen was detected approximately 17 cm from the preputial orifice. Reconstructive surgery was the chosen therapy, with amputation of the affected foreskin portion. A skin segment of about 12 cm and affected mucosal portion were removed. Postoperative therapy consisted of dressing with chlorhexidine digluconate based ointment, povidone iodine and topical insect repellent (cypermethrin and carbamyl), systemic antibiotic therapy with benzathine penicillin (30,000 IU / kg, repeated after 48 hours) and anti-inflammatory therapy with flunixin meglumine (2 , 0 mg / kg SID) for three days. After twenty days, semen was collected by massage of the accessory glands. The ejaculate volume was 3 mL, with 200x106 spermatozoa / mL, white color, "suis generis" odor, 88% of motile sperm, 64% of progressively motile sperm and 82% of rapid sperm. Thus, the chosen surgical treatment was effective to correct the pathology and reintroduce the animal into reproductive activity

Termografia digital por infravermelho do escroto e qualidade do sêmen em touros nelore (bos taurus indicus)

Aimed to study the relationship between scrotal temperature on semen quality by use of infrared thermography in Nelore bulls bred extensively. There were scrotal thermography and semen collection. Climatic data were collected by means of globe thermometer. The thermograms of the scrotum were obtained by digital infrared thermography. We carried out ejaculates through electroejaculation for analysis of quantitative and qualitative characteristics. For the data, we used analysis of variance and subsequently applied the Tukey test at 5%. (p <0.05) between animals for sperm vigor, sperm motility between animals and between crops, sperm concentration, total sperm and total viable sperm between groups, between harvests for the scrotum temperatures and climate data. It was concluded that the surface temperatures of the scrotum and climatic factors influence the quality of semen. Thermography is recommended as a complementary test in the evaluation of reproductive bulls.

Comparison of apoptotic cells between cryopreserved ejaculated sperm and epididymal sperm in stallions

The development of a reliable technique to freeze epididymal semen would provide a unique opportunity to preserve valuable genetic material from unexpectedly lost stallions. The aim of this study was to compare the apoptotic indices of sperm obtained from ejaculate, sperm recently recovered from the epididymides (EP), and sperm recovered from epididymides stored at 5 C for 24 hours (EP-stored). For the first category, two ejaculates from seven stallions were collected and then submitted to cryopreservation using an egg yolk-based extender. One week after the last semen collection, the stallions were submitted to bilateral orchiectomy, and sperm from one of the cauda epididymis was harvested immediately after castration (EP). The remaining testicle was stored in a passive refrigeration container at 5 C for 24 hours before the cauda epididymal sperm was harvested (EP-stored). Sperm harvesting from the epididymis for EP and EP-stored was performed by retrograde flushing of the caudal portion of the epididymis using a skim milk-based extender. The recovered sperm was then cryopreserved using the egg yolk-based extender. Sperm motility parameters were studied by computerassisted semen analysis, and apoptosis was estimated by measuring caspase activity and membrane phospholipid translocation using epifluorescence microscopy. The samples were evaluated immediately (0 hour) and 8 hours after thawing. At 0 hour, no differences in sperm parameters were observed among the groups, but after 8 hours, significant statistical differences were observed in sperm motility parameters and plasma membrane integrity among the treatment groups. In addition, viable cells with no apoptotic signs were more prevalent in EP and EP-stored, suggesting that epididymal sperm is less sensitive to the cold shock caused by sperm cryopreservation.

Efeito do período pós-parto sobre a taxa de prenhez de vacas de corte submetidas à IATF (Inseminação Artificial em Tempo Fixo)

This work aimed to evaluate the influence of postpartum period (precocious - of 28 to 44 days and late - of 45 to 90 days) on the bovine pregnancy rate using fixed-time AI. For that, 678 cows were divided in two groups: precocious group (G-P, n=151) and late group (G-T, n=527). The animals received CIDR® + 2 mL of estradiol benzoate (BE) in the day zero (D0). After eight days (D8) the dispositive was removed and both groups received 2.5mL PGF2α, concurrent with PGF2α injection, they received either 1.5mL of eCG or temporary calf removal (RTB). In the next day (D9), the cows received 1 mL de BE and 24 hours later, the fixed-time AI was performed with Nelore bovine semen. The calves were returned to their mothers. The pregnancy rate was not different between the groups (p>0.05), G-P=40% and G-T=48%. The results indicate that females with less than 45 days of postpartum are able to hormone protocol of fixed-time IA.

Correlação da circunferência escrotal, citologia testicular e parâmetros do sêmen do epidídimo de touros

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Efeito da adição de butil-hidroxitolueno nos meios de refrigeração e congelação sobre a viabilidade espermática de equinos

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Soroprevalência, isolamento e diagnóstico molecular para leptospirose em bovinos de fazendas de cria e de centrais de receptoras de embrião da região de Bauru- SP: Diagnóstico de leptospirose bovina em Bauru - SP

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Estudo sobre a redução do estresse oxidativo em sêmen equino a partir da adição de quercetina nos diluentes de refrigeração e congelação

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Efeito da centrifugação e filtragem do sêmen bovino sobre a criopreservação espermática

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Estudo sobre a redução do estresse oxidativo em sêmen equino a partir da adição de quercetina nos diluentes de refrigeração e congelação

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)