531 resultados para Semen - Criopreservação


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

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Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

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Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

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The present study evaluated the use of PCR for Histophilus somni detection in bovine semen. Semen samples were experimentally infected with H. somni at dilutions ranging from 107 to 101 bacteria/mL and subjected to DNA extraction by the phenol/chloroform method, followed by PCR amplification. The amplification products were analyzed by electrophoresis in 8% acrylamide gel. The oligonucleotide primers used yielded an amplification fragment of 400 base pairs from the bacterial DNA. Positive amplification was obtained even for the 101 bacteria/mL dilution. PCR proved to be an efficient method for the detection of H. somni. The results obtained in this study have brought relevant information for the diagnosis of H. somni, justifying the need for the diagnosis of this bacterium in bulls, especially in semen samples that should be free of contamination. The PCR method has shown to be a useful tool for the quality control of semen produced in artificial insemination centers.

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Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

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Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

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Brucellosis is a zoonosis caused by bacteria of the genus Brucella. Man infection occurs through contact with reproductive secretions as placenta and its lochia, semen and penile secretion of infected animals or by consuming unpasteurized milk and dairy products. With the objective of investigating the presence of bacteria in milk, 30 samples of raw milk sold illegally in the region of Botucatu, São Paulo, Brazil, as well as 50 samples of milk delivered to a dairy industry previously to its pasteurization were evaluated by the polymerase chain reaction (PCR) technique. Of the 80 samples analyzed, 10 samples (12.5%) were positive and 70 (87.5%) were negative. Among the  positive samples,  5 (16.6%)  were from  illegal traders  and other  5  (10%) were obtained  from the dairy industry. Brucella spp. positivity shows that the pathogen is representatively present in Botucatu, São Paulo, Brazil, and the risk associated to public health due to the commercialization of illegal products without pasteurization is real.

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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.

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

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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.

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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

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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.