The objective of the present study was to describe the relationship of seminal plasma and total sperm cell proteins with the semen freezability parameters of Guzerat bulls. Thirteen bulls were subjected to breeding soundness evaluation. Semen samples were collected, cryopreserved, and then post-thawing sperm kinetics were assessed, where high ( = 7) and low ( = 6) freezability groups were defined. Seminal plasma and total sperm proteins from the 2 groups were separated by 2-dimensional SDS-PAGE, and spots were identified by mass spectrometry. Semen parameters post-cryopreservation were as follows in the high and low freezability groups, respectively: mean total motility, 52.4 ± 20.5 and 13.7 ± 3.9; percentage of normal sperm, 89.0 ± 2.6 and 64.7 ± 14.0; and reactivity of hypo-osmotic swelling test, 38.9 ± 4.7 and 13.6 ± 3.7. Three seminal plasma proteins (osteopontin-K, DNase γ precursor, and DNASE1L3) and 6 proteins from sperm cells (acrosome formation-associated factor isoform 2, annexin A1, disintegrin and metalloproteinase domain-containing protein 2, dihydrolipoyl dehydrogenase, and glyceraldehyde-3-phosphate dehydrogenase) were highly expressed ( < 0.05) in the high freezability group. Another 6 seminal plasma proteins (acrosin inhibitor 1, glutathione peroxidase 3, metalloproteinase inhibitor 2, ephrin-A1, annexin A1, and platelet-activating factor acetylhydrolase) were significantly higher ( < 0.05) in the low freezability group. We described the associations of seminal plasma and sperm cell proteins with post-thawing sperm viability of Guzerat bulls raised in a typical semiarid environment. Such associations indicate that specific seminal plasma proteins more abundant in bulls of low semen freezability may be a response to an early oxidative stress that is not detected by conventional prefreezing semen evaluation. Moreover, specific sperm proteins were more associated with good freezability. The results presented here can serve as guidelines for future research aiming to develop better extenders and/or to improve bull semen selection for cryopreservation.

Resumo -O objetivo deste trabalho foi avaliar a presença do DNA pró-viral do lentivírus caprino (LVC) em ejaculados de machos infectados naturalmente, e verificar a influência da lavagem do sêmen e da presença de inflamação testicular sobre a carga viral. Foram realizadas oito coletas de sêmen de sete reprodutores soropositivos para o LVC: quatro antes dos animais sofrerem dano testicular e quatro depois. Entre as coletas realizadas na mesma semana, em uma, o ejaculado era lavado, para retirada do plasma seminal, e na outra, não. O DNA pró-viral do LVC foi identificado pela reação em cadeia da polimerase Nested (PCR Nested), e pelo isolamento viral. O vírus foi isolado em 7,1% das amostras. A PCR identificou o DNA pró-viral em 35,7% do total das amostras: 17,9% nas amostras lavadas e 53,6% das amostras de sêmen integrais. O dano ao testículo permite maior fluxo do vírus para o sêmen, pois antes do dano, 21,4% das amostras foram positivas e pós-dano, 50%. A transmissão do LVC pelo sêmen de reprodutores caprinos é potencializada pela presença de inflamações testiculares e pelo fato de o sêmen criopreservado conter o LVC na forma infectante.Termos para indexação: reprodução, PCR, artrite-encefalite caprina, transmissão. Risk factors in caprine lentivirus transmission through semenAbstract -The objective of this work was to evaluate the presence of the DNA provirus of the caprine lentivirus (LVC) in ejaculates of naturally infected males, and to verify the influence of the wash of the semen as well as the presence of testicle inflammation on the viral load. Eight semen collections of seven soropositive reproducers were accomplished, four before testicle injury and four after injury. Amongst the collections carried out at the same week, in one the ejaculate was washed, to withdraw the plasma seminal, and in the other it was not. The provirus DNA was identified both by Nested polymerase chain reaction technique (Nested PCR) and by the viral isolation. The virus was isolated in 7.1% of the samples. The PCR identified the provirus DNA in 35.7% of all samples, 17.9% in the washed samples and 53.6% of the integral semen samples. The injury of the testicle tends to greater flow of virus for the semen, therefore, before injury, 21.4% of the samples were positive and after-injury, 50%. Risk of transmission of the LVC by semen of goat reproducers is strengthened by the presence of testicle inflammations and the fact that the criopreserved semen contains the LVC in infecting form.Index terms: reproduction, PCR, caprine arthritis encephalitis, transmission. IntroduçãoO lentivírus caprino (LVC) é o agente etiológico da artrite-encefalite caprina (AEC), uma enfermidade crônica, incurável, de alta prevalência em rebanhos leiteiros nacionais e associada à perdas econômicas. Há grande necessidade de pesquisa dos modos de transmissão desse vírus para o delineamento eficiente de medidas de controle e possível erradicação.O avanço das técnicas de conservação de germoplasma, aliado ao aumento da importância econômica da caprinocultura, ...

Ejaculados de seis bodes Moxotó e seis ½ sangue Moxotó-Pardo Alpina, adultos, mantidos em confinamento, foram avaliados antes e após a insulação do saco escrotal com bolsa de plástico, de parede dupla, durante 6,5 dias. A degeneração seminal ocorreu em todos os animais na 4ª semana após o início da insulação, ressaltando-se a redução na concentração, o aumento dos defeitos espermáticos e a redução do vigor celular, culminando com a necrospermia. A motilidade individual progressiva (MIP) atingiu os valores mais baixos na 3ª semana após o início da insulação, retornando aos valores normais entre a 8ª e 9ª semanas. Os defeitos espermáticos começaram a aumentar aos sete dias após o início da insulação escrotal. O volume apresentou oscilações atípicas e o aumento da temperatura escrotal afetou os parâmetros físicos e morfológicos do ejaculado de maneira consistente evidenciando-se o efeito negativo sobre a qualidade do sêmen.

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Background: Caprine Arthritis Encephalitis Virus have been detected in sperm of breeding goats causing economic losses. In order to control the virus, researches aiming to identify natural extracts with potential antiviral effects are performed. However, aqueous or ethanolic extracts must be diluted in dimethyl sulfoxide (DMSO), which is a substance with unknown effects in sperm quality when present in diluting media. Therefore, this study aimed to evaluate sperm viability of refrigerated caprine semen diluted in media containing DMSO. This was performed to provide data that aid in researches involving the use of this component with natural extracts that may inactivate the caprine lentivirus in sperm.Materials, Methods & Results: The experiment was performed at the Laboratory of Seminal Technology in Embrapa Goats and Sheep in the city of Sobral, Brazil. Sperm viability was assessed in caprine semen refrigerated in two dilution media with crescent concentrations of DMSO. Sperm samples of five goats seronegative for the caprine lentivirus were pooled and diluted in minimal essential medium (MEM) enriched with glucose at 0.01 M added of crescent concentrations of DMSO (0%, 1.5%, 1.75%, 2.0%, 2.25% and 2.5%). The same breeders provided the pool of sperm to test Tris added 2.5% of egg yolk and the same concentrations of DMSO previously mentioned. Treatments were refrigerated at 7°C and evaluated up until four h after DMSO addition. Individual progressive motility (MIP), sperm vigor (V), percentage of spermatozoa reactive to hypoosmotic test (HO) and morphologically normal (NOR) were evaluated. IPM, vigor and NOR remained within normal standards for the caprine species in all treatments test. Percentage results of spermatozoa reactive to hypoosmotic was higher in Tris yolk with values ranging between 34.66% to 46.33%. Sperm vigor was positively correlated (r = 0.85) with IPM in the MEM diluted pool of sperm. In Tris yolk, vigor and hypoosmotic test correlated moderately (r = 0.63, r = 0.54, respectively) with IPM. Tris yolk medium added DMSO presented the highest percentage of reactivity to hypoosmotic test in all treatments when compared to MEM added DMSO.Discussion: The fact that DMSO is easily homogenized in water, ethylic alcohol and most organic solvents favors its use in diluting natural extracts. These components are a possible source of products that inactivate caprine arthritis encephalitis virus in sperm, which is the key to promoting the safe use of genetic material of infected breeders, in addition to commercial use of germplasm. In this study, there was no interference of DMSO in the analyzed parameters when added in a maximum concentration of 2.5% to MEM and Tris yolk, which is in accordance with standard values for goats. In addition, Tris yolk may promote greater protection to the membrane of sperm cells, which was demonstrated by hypoosmotic test. This medium could be ideal to be used in new methodologies that incorporate DMSO. In conclusion, DMSO added to dilution media Tris yolk and MEM did not interfere with the quality of refrigerated caprine sperm, which maintained viability. These results indicate that this substance did not present harmful effects to the genetic material, promoting the use as solvent of extracts from plant compounds with potential anti-viral effect. The information in this study may aid new research performed in this area.