The presence and gene expression of bovine papillomavirus in the peripheral blood and semen of healthy horses.

Papillomavirus (PV) are double-stranded DNA viruses that can cause both benignant and malignant tumours in mammals. Twelve genotypes of bovine papillomavirus (BPV1-12) have been identified so far. The presence of BPV1 and 2 has been found in the body fluids of cattle and horses. The aim of this study is to investigate the presence of BPV DNA and the expression of viral genes in the blood and sperm cells of healthy horses using PCR and RT-PCR. BPV-1 or 2 was detected in 14 of 70 blood samples (20%) and in 11 of 31 semen samples (35%). In five of fourteen blood samples, the E5 expression tested positive, while no blood sample was positive for L1 expression. Four of 11 (36%) semen cell samples proved to be positive for E5 expression, while no gene expression in L1 could be detected. This is the first study that shows BPV1 gene expression in the blood and semen of healthy horses. Our data illustrate the need for a better understanding of the presence of BPV in non-epithelial tissues of horses and their role in the vertical and horizontal transmission of these viruses.

Detection of bovine papillomavirus type 2 DNA in commercial frozen semen of bulls (Bos taurus).

Papillomaviruses are found in epithelial lesions and are linked to different carcinogenic processes in humans and other animals. Although bovine papillomavirus (BPV) has been characterized as epitheliotropic, the presence of viral DNA has been detected in other sample types, including fresh semen. The aim of this study was to evaluate the presence of BPV DNA in spermatozoa and seminal plasma samples of commercial frozen semen taken from bulls (Bos taurus) and its effects on semen function. PCR assays were conducted with specific primers to detect BPV types 1-6 in 40 semen samples of dairy Gir bulls. The semen quality was assessed by the use of parameters such as motility, vigor, acrosomal integrity and DNA integrity. BPV-2 DNA was detected in all of the sperm cell samples and all the seminal samples; however BPV-1, 3, 4, 5 and 6 could not be detected. The presence of BPV DNA was apparently not a cause of reduced sperm function. This is the first record of BPV-2 DNA the commercial frozen semen taken from dairy Gir cattle by several companies that provide semen. Further studies are needed to assess the viability of the virus and the extent to which it can be spread through semen.

Efeito da adição de trolox e pentoxifilina na motilidade, integridade do acrossoma e do DNA de espermatozoides equinos após descongelação / Effect of trolox and pentoxifylline on motility and integrity of, acrossome and DNA of equine spermatozoa after thawing

Três garanhões foram utilizados para estudar o efeito da adição de trolox e pentoxifilina na motilidade, integridade do acrossoma e DNA de espermatozoides pós-descongelação. Para congelação, utilizou-se Tris-gema com glicerol (5 por cento) em máquina de congelação de sêmen. As amostras foram descongeladas a 37ºC durante 30 segundos e tratadas com: T1= 150µL de sêmen + 150µL de Tris; T2= 150µL de sêmen + 150µL de Tris + 120µM/mL de trolox; T3= 150µL de sêmen + 150µL de Tris + 3,5mM de pentoxifilina e T4= 150µL de sêmen + 150µL de Tris + 3,5mM de pentoxifilina + 120µM/mL de trolox. Após 0, 60 e 120 minutos de incubação (37ºC), as amostras foram analisadas quanto à motilidade, vigor, integridade de acrossoma e DNA. Não houve diferença (P>0,05) entre tratamentos após 0 e 60 minutos de incubação em todos os parâmetros estudados. Após 120 minutos de incubação, verificou-se maior porcentual (P<0,05) de células com motilidade total e progressiva nas amostras do T2. Conclui-se que a adição de trolox após descongelação do sêmen equino preserva a motilidade total e progressiva dos espermatozoides submetidos à incubação a 37ºC durante 120 minutos.

Three stallions were used to study the effect of trolox and pentoxifylline addition on the motility and integrity of acrossome and DNA equine spermatozoa after thawing. Tris-egg-yolg diluent with glycerol (5 percent) were used to freeze the semen samples in a freezing machine. The samples were thawed at 37ºC during 30 seconds and treated with: T1=150µL of semen + 150µL of Tris; T2= 150µL of semen + 150µL of Tris +150mM/mL of trolox; T3= 150µL of semen + 150µL Tris +3.5mM of pentoxifylline; and T4= 150µL of semen + 150µL of Tris + 3.5mM of pentoxifylline + 150mM of trolox. After 0, 60, and 120 minutes of incubation (37ºC), the samples were analyzed to motility, vigor, and integrity of acrossome and DNA. There was no difference (P>0.05) among treatments considering 0 and 60 minutes of incubation in all studied parameters. After 120 minutes of incubation, it was observed higher percentage (P<0.05) of cells with total and progressive motility in the samples of T2. It can be concluded that the trolox addition after thawing of equine semen preserved total and progressive motility of the sperm incubated at 37ºC during 120 minutes.