Short Communication: Comparative Susceptibility of Rhesus Macaques of Indian and Chinese Origin to Vaginal Simian-Human Immunodeficiency Virus Transmission as Models for HIV Prevention Research.

Historically, Indian rhesus macaques (iRMs) have been preferred for simian immunodeficiency virus (SIV)/HIV prevention, pathogenesis, and treatment studies, yet their supply is limited. Chinese rhesus macaques (cRMs) are currently more available, yet little is known regarding the relative susceptibility of this subspecies to vaginal transmission of SIV or simian-human immunodeficiency virus (SHIV). In this study, we compared the susceptibility of 40 cRMs and 21 iRMs with a single vaginal challenge with SHIVsf162P. Our results showed that cRMs have comparable primary SHIV infection as iRMs, underscoring their equal importance in studies of HIV transmission and prevention.

The Major Envelope Glycoprotein of Murid Herpesvirus 4 Promotes Sexual Transmission.

Gammaherpesviruses are important human and animal pathogens. Infection control has proven difficult because the key process of transmission is ill understood. Murid herpesvirus 4 (MuHV-4), a gammaherpesvirus of mice, is transmitted sexually. We show that this depends on the major virion envelope glycoprotein gp150. gp150 is redundant for host entry, and in vitro, it regulates rather than promotes cell binding. We show that gp150-deficient MuHV-4 reaches and replicates normally in the female genital tract after nasal infection but is poorly released from vaginal epithelial cells and fails to pass from the female to the male genital tract during sexual contact. Thus, we show that the regulation of virion binding is a key component of spontaneous gammaherpesvirus transmission.IMPORTANCE Gammaherpesviruses are responsible for many important diseases in both animals and humans. Some important aspects of their life cycle are still poorly understood. Key among these is viral transmission. Here we show that the major envelope glycoprotein of murid herpesvirus 4 functions not in entry or dissemination but in virion release to allow sexual transmission to new hosts.

Occurrence of equine coital exanthema (ECE) in stallions in Japan and effectiveness of treatment with valacyclovir for ECE.

Equine coital exanthema (ECE) has been reported in many countries, but equine herpesvirus 3 (EHV-3) has been isolated only once in Japan. In 2015, symptoms of ECE were found, and EHV-3 was isolated in two stallions. Valacyclovir, an anti-herpesvirus agent, was administered orally. The stallions rested from mating for more than two weeks, causing enormous financial losses because of their high fees. This is the first study in which valacyclovir was administered for ECE. Though valacyclovir treatment did not shorten the duration of healing, the affected area did not expand after administration of valacyclovir. Valacyclovir therefore seems to be effective for suppression of EHV-3 infection. Further investigation about the administration protocol might be required.

Isolation of equine herpesvirus 3 (EHV-3) from equine coital exanthema of two stallions and sero-epidemiology of EHV-3 infection in Japan.

In the spring of 2015, two stallions reared in Farms A and B in Hokkaido in Japan showed symptoms of equine coital exanthema. Equine herpesvirus 3 (EHV-3) was isolated from penis swab samples of both stallions, and the isolates from each stallion in Farms A and B were designated as SS-1 and YS-1 strains, respectively. BamHI restriction profiles of SS-1 and Japanese reference strain Iwate-1 were indistinguishable, but the BamHI-A fragment of YS-1 was larger than those of SS-1 and Iwate-1 by 1.9 kbp because of the lack of two BamHI sites. Nucleotide sequence analyses of glycoprotein G (gG), gB, gC and VP13/14 coding regions revealed that SS-1 and YS-1 had 99.77% to 100% identities to each other. These results suggested that the origins of SS-1 and YS-1 were different. For a sero-epidemiological survey, serum neutralizing tests using SS-1 against 319 sera of horses from eight farms in Hokkaido were conducted. Six of the eight farms were EHV-3 antibody-positive, and positive rates ranged from 2.6% to 17.6%. To determine the infection time of four EHV-3 antibody-positive horses, a retrospective study was conducted. Infection time of the four horses was in the breeding season, and re-infection or reactivation of latently infected EHV-3 might have occurred in one horse. However, these four horses had never shown any clinical symptoms. The results suggested that several EHV-3 strains are distributed in Japan and that infection is maintained widely in horses without clinical symptoms.

Detection of Leishmania infantum in the smegma of infected dogs / Detecção de Leishmania infantum no esmegma de cães infectados

Considering the venereal transmission of visceral leishmaniasis from dogs to bitches, the aim of this study was to verify if the penile surface and smegma from infected dogs can be the source of parasites in bitches. Twelve Leishmania infantum infected dogs had semen and smegma samples collected for submission to PCR identification of the DNA of the parasite. Semen (41.7 percent) and smegma (50.0 percent) have similar positive incidence (P>0.05; Fisher's exact test), with 58.3 percent of the dogs positive for semen and/or smegma samples. The proportion of positivity for both semen and smegma was 33.3 percent, but 8.3 percent was positive only for semen, and 16.7 percent only for smegma, revealing a moderate agreement between tests (K=0.5; Kappa index). It was concluded that Leishmania infantum is present in the smegma of contaminated dogs and it can be a source of parasites for the semen and the bitch...

Tendo em vista a transmissão venérea da leishmaniose visceral do cão para a cadela, o objetivo deste estudo foi verificar se a superfície peniana e o esmegma de cães infectados poderiam ser a fonte de parasitas para a fêmea. Amostras de sêmen e esmegma de 12 cães infectados com Leishmania infantum foram submetidas à identificação do DNA do parasita por PCR. As incidências de positividade no sêmen (41,7 por cento) e no esmegma (50,0 por cento) foram semelhantes (P>0,05; teste exato de Fisher), sendo 58,3 por cento dos cães positivos para sêmen e/ou esmegma. A positividade para sêmen e esmegma juntos ocorreu em 33,3 por cento, mas em 8,3 por cento dos casos apenas no sêmen, e em 16,7 por cento apenas no esmegma, o que revela uma concordância moderada entre os testes (K=0,5; índice Kappa). Conclui-se que a Leishmania infantum está presente no esmegma de cães contaminados, podendo ser a fonte de parasitas para o sêmen e a cadela...

Evaluation of the transmission of porcine circovirus type 2 (PCV-2) genogroups a and b with semen from infected specific-pathogen-free boars.

The porcine circovirus type 2 (PCV-2) is associated with several diseases including reproductive failure. This syndrome has been experimentally reproduced twice with two PCV-2 isolates representative of each major PCV-2 genogroup, i.e. PCV-2a and PCV-2b (Cariolet et al., 2002; Rose et al., 2007). In these two previous studies, the sows were infected by intra-uterine inoculation at insemination with 10(4.3) and 10(3.18) TCID(50) of PCV-2a and PCV-2b, respectively, corresponding to 1.2 × 10(11) and 3 × 10(10) genome copies, respectively. The aim of this present study was to quantify viral shedding in semen from specific-pathogen-free (SPF) boars infected with isolates from the two major PCV-2 genogroups a and b. We studied the transmission of the PCV-2 virus through contaminated semen to SPF sows and their offspring. The four inoculated boars developed sub-clinical PCV-2 infections and PCV-2 genomes were occasionally detected in semen after nasal infection of boars, with up to 1.2 × 10(6)copies/mL in the sperm-rich fraction. When PCV-2-contaminated semen was inoculated in SPF sows at artificial insemination, the sows and their offspring did not show any signs of PCV-2 infection or PCV-2 antibodies or genomes. In the present study, sows were inoculated with a maximal dose of 1.7 × 10(7) viral genome copies, which is lower than the genomic loads (i.e. 1.2 × 10(11) and 3 × 10(10) genome copies) that have been shown to induce reproductive troubles in intra-uterine inoculated sows. Our results together with the previous experiment findings suggest that PCV-2-induced reproductive disorders depend on the infectious dose inoculated to sows by the intra-uterine route.

Preferential sexual transmission of pseudorabies virus in feral swine populations may not account for observed seroprevalence in the USA.

This paper compares the behavior of two competing models for the transmission of pseudorabies virus in feral swine in the USA. In first model, horizontal (non-sexual) density dependent transmission is the only transmission modality. In the second model, the only transmission modality is sexual transmission between mature males and females. The comparison of model behavior was carried out to test the hypothesis that preferential sexual transmission of PRV in feral swine can account for the seroprevalence observed in the field. The observed range of seroprevalence of PRV in mature feral swine in the USA is consistent with a preferential sexual transmission only if the feral swine mating system is a random mating system or a polygynous system in which there is a relatively large rate of acquisition of new mates. The observed range of seroprevalence of PRV in mature feral swine in the USA is not consistent with a preferential sexual transmission if there is mate guarding. This is important because the National Pseudorabies Surveillance Plan deems monitoring the risk of PRV introduction from feral swine to be a "minor objective" both in terms of the scope of the plan and with respect to the resources allocated. The rationale for this statement was derived from experimental studies, which suggested that the PRV indigenous to feral swine in the USA is preferentially sexually transmitted.

Infectious bronchitis virus in testicles and venereal transmission.

Even though males represent only 8%-12% of the birds of a breeder flock, their role in infectious bronchitis virus (IBV) dissemination is largely unknown. We first assessed the effect of IBV replication in the chicken testes. Ten-week-old males were inoculated with Arkansas (Ark) or Massachusetts (Mass) IBV virulent strains. Seven days postinoculation (DPI) IBV RNA was detected in testicles of 100% of M41- and in 96% of Ark-infected males. Marginal nonsynonymous variation was detected in spike (S) gene of the predominant population of IBV replicating in the testes compared to the S gene of the predominant population of viruses prior to inoculation. IBV M41 and Ark were detected in spermatogonia and Sertoli cells of testicles of infected roosters by immunofluorescence, without evident histopathological changes. We next assessed venereal transmission of IBV by artificially inseminating 54-wk-old hens either with semen from IBV-infected roosters or with IBV suspended in naïve semen. IBV RNA was detected in the trachea of all hens inseminated with IBV-spiked semen and in 50% of hens inseminated with semen from IBV-infected males. The egg internal and external quality was negatively affected in hens inseminated with semen containing IBV. These results provide experimental evidence for IBV venereal transmission.

Equine viral arteritis: current status and prevention.

Recently, there has been increased interest in equine viral arteritis (EVA) among veterinarians and horse owners. Outbreaks of the disease were identified initially in New Mexico, USA in 2006, and in the Normandy region of France in the summer of 2007. Both occurrences were associated with AI of cool-shipped semen. Each was linked to respiratory illness, neonatal death, abortion, development of carrier stallions, and cancellation of equestrian events. In light of the increased interest, this paper will present a brief case history, followed by a review addressing common concerns regarding EVA, current status, and control and prevention strategies, including vaccination, and recommended bio-security measures.

Venereal and vertical transmission of deformed wing virus in honeybees (Apis mellifera L.).

Deformed wing virus (DWV) infected semen was used for artificial insemination of DWV-free virgin queens. High titres of DWV could subsequently be detected not only in the spermatheca, but also in the ovaries, demonstrating venereal transmission of DWV in honey bees. Subsequent vertical transmission of the virus to the progeny of DWV infected queens was also demonstrated. Neither transmission route is 100% effective. Whether venereal transmission of DWV occurs during natural mating remains to be determined. The implications for the use, sale and transport of semen samples for artificial insemination are discussed.

Models for an arenavirus infection in a rodent population: consequences of horizontal, vertical and sexual transmission.

Arenaviruses are associated with rodent-transmitted diseases in humans. Five arenaviruses are known to cause human illness: Lassa virus, Junin virus, Machupo virus, Guanarito virus and Sabia virus. In this investigation, we model the spread of Machupo virus in its rodent host Calomys callosus. Machupo virus infection in humans is known as Bolivian hemorrhagic fever (BHF) which has a mortality rate of approximately 5-30% [31]. Machupo virus is transmitted among rodents through horizontal (direct contact), vertical (infected mother to offspring) and sexual transmission. The immune response differs among rodents infected with Machupo virus. Either rodents develop immunity and recover (immunocompetent) or they do not develop immunity and remain infected (immunotolerant). We formulate a general deterministic model for male and female rodents consisting of eight differential equations, four for females and four for males. The four states represent susceptible, immunocompetent, immunotolerant and recovered rodents, denoted as S, I( t), I( c)and R, respectively. A unique disease-free equilibrium (DFE) is shown to exist and a basic reproduction number R( 0)is computed using the next generation matrix approach. The DFE is shown to be locally asymptotically stable if R(0) < 1and unstable if R( 0) > 1. Special cases of the general model are studied, where there is only one immune stage, either I(t) or I(c). In the first model, SI(c)R( c), it is assumed that all infected rodents are immunocompetent and recover. In the second model, SI(t), it is assumed that all infected rodents are immunotolerant. For each of these models, the basic reproduction numbers are computed and their relationship to the basic reproduction number of the general model determined. For the SI( t)model, it is shown that bistability may occur, the DFE and an enzootic equilibrium, with all rodents infectious, are locally asymptotically stable for the same set of parameter values. A simplification of the SI( t)model yields a third model, where the sexes are not differentiated, and therefore, there is no sexual transmission. For this third simplified model, the dynamics are completely analyzed. It is shown that there exists a DFE and possibly two additional equilibria, one of which is globally asymptotically stable for any given set of parameter values; bistability does not occur. Numerical examples illustrate the dynamics of the models. The biological implications of the results and future research goals are discussed in the conclusion.

Age-prevalence of Otarine Herpesvirus-1, a tumor-associated virus, and possibility of its sexual transmission in California sea lions.

Otarine Herpesvirus-1 (OtHV-1) is a gammaherpesvirus routinely detected in urogenital tumor tissues of adult sea lions dying during rehabilitation, To investigate the epidemiology of this virus and guide the development of a mathematical model of its role in the multifactorial etiology of cancer in California sea lions, polymerase chain reaction (PCR) amplification of an OtHV-1 specific fragment of the DNA polymerase gene was used to look for evidence of OtHV-1 infection in urogenital and pharyngeal swabs and peripheral blood mononuclear cells (PBMC) of sea lions of different ages. Samples were also examined from pregnant females and their late term in utero or aborted fetuses to investigate potential for vertical transmission. Prevalence of infection in 72 adult females was 22%, whereas it was 46% in 52 adult males, and was significantly lower in 120 juvenile animals (6%). OtHV-1 DNA was most often detected in the lower reproductive tract of the adult animals, especially the males, and rarely in the pharynx or urogenital tract of juvenile animals. These data suggest sexual transmission may an important route of transmission. Additional studies are required to confirm this mode of transmission. Additionally, the virus was detected in a single prematurely born pup, suggesting the possibility of perinatal transmission. No indication of a PBMC associated viremia was evident in adults using standard PCR or in juveniles using standard and real time PCR.

Disease transmission in horses.

Bacterial, viral and protozoal infections may cause severe reproductive losses. The present paper reviews the risk factors, clinical signs and preventive measures for the most important venereal or potential sexually transmitted diseases in horses. The stallion and use of semen for artificial insemination represent major risk factors for the transmission of bacterial contaminants of the penis, including Streptococcus equi subspecies zooepidemicus, Pseudomonas aeruginosa and Klebsiella pneumoniae, known to cause endometritis and infertility in the mare. The role of the stallion in disease transmission is also due to the non-clinical manifestation of diseases such as contagious equine metritis and equine viral arteritis. Dourine has been eradicated from many countries, but continues to be a problem in other areas of the globe. Strategies for the prevention of introduction and transmission of diseases in breeding operation are discussed.

Occurrence of equine coital exanthema in pastured draft horses and isolation of equine herpesvirus 3 from progenital lesions.

During the period from 2001 to the following year, progenital diseases had been epidemic among the draft stallions and mares pastured together in Iwate Prefecture, the northeastern district of Japan. A stallion and 8 of 31 mares were affected in 2001, and 1 of 2 stallions and 10 of 36 mares in 2002. The clinical symptoms consisted of the formation of papules, pustules, ulcers and scabs on the progenital skin and mucosa in stallions and mares. In 2002, Equine herpesvirus 3 (EHV3) was isolated from 2 mares and the glycoprotein G gene of the virus detected from a stallion and 4 mares by polymerase chain reaction. Serum neutralizing tests showed that 12 of 38 horses, 10 clinically and 2 subclinically affected, changed to be positive for the EHV3 antibody. The results suggest that the horses were affected with equine coital exanthema (ECE) through coitus. Five mares with the antibody at the pre-pastured period may have been the possible origins of EHV3 infection in 2002, although the exact origin in 2001 remains unknown. The artificial insemination was performed for the prevention of ECE spreading through coitus on the pasture in 2003. There was no epidemic of the disease in 31 mares, although 3 mares with the antibody at the pre-pastured period showed the significant increase in the titers during the pastured period.

Potential sites of virus latency associated with indigenous pseudorabies viruses in feral swine.

Free-ranging feral swine (Sus scrofa) are known to be present in at least 32 states of the USA and are continuously expanding their range. Infection with pseudorabies virus (PRV) occurs in feral swine and the primary route of transmission in free-living conditions seems to be venereal. Between 1995 and 1999, naturally infected feral swine and experimentally infected hybrid progeny of feral and domestic swine, were kept in isolation and evaluated for occurrence of latent PRV indigenous to feral swine in sacral and trigeminal ganglia and tonsil. Sacral ganglia were shown, by polymerase chain reaction (PCR) amplification of the thymidine kinase (TK) gene of PRV, to be the most frequent sites of latency of PRV. Nine (56%) of 16 sacral ganglia, seven (44%) of 16 trigeminal ganglia, and five (39%) of 13 tonsils from naturally infected feral swine were positive for PCR amplification of TK sequences of PRV. These tissues were negative for PRV when viral isolation was attempted in Vero cells. DNA sequencing of cloned TK fragments from the sacral ganglia of two feral swine, showed only one nucleotide difference between the two fragments and extensive sequence homology to fragment sequences from various domestic swine PRV strains from China, Northern Ireland, and the USA. The hybrid feral domestic swine, experimentally inoculated with an indigenous feral swine PRV isolate by either the genital or respiratory route, acquired the infection but showed no clinical signs of pseudorabies. Virus inoculated into either the genital or respiratory tract could, at times, be isolated from both these sites. The most common latency sites were the sacral ganglia, regardless of the route and dose of infection in these experimentally infected hybrids. Nine of 10 sacral ganglia, six of 10 trigeminal ganglia, and three of 10 tonsils were positive for PCR amplification of TK sequences. No virus was isolated from these tissues in Vero cells. The demonstration of the sacral ganglia as the most common sites of latency of pseudorabies viruses indigenous to feral swine, supports the hypothesis that these viruses are primarily transmitted venereally, and not by the respiratory route as is common in domestic swine, in which the trigeminal ganglia are the predominant sites of virus latency.

Localization of porcine reproductive and respiratory syndrome virus infection in boars by in situ riboprobe hybridization.

The capability of porcine reproductive and respiratory syndrome virus (PRRSV) to be shed in semen for extended periods of time has been suggested to be a principal factor for viral transmission via insemination. In attempts to gain insights into the mechanism of PRRSV persistence in boars, tissue distribution and sites of viral infection were investigated by in situ hybridization (ISH) using digoxigenin-labeled RNA probe and the ISH results were compared with those of reverse transcription-nested polymerase chain reaction (RT-nested PCR). Animals were intranasally inoculated with 10(4) median tissue culture infectious dose of PRRSV VR-2332 and tissues collected at different times were examined. At day 7 postinfection, limited number of hybridization positive signals was observed in cells within or between seminiferous tubules in the testis sections while relatively abundant hybridization positive signals were observed in the brain stem and tracheobronchial lymph node. At later days of infection, hybridization positive signals were observed in cells within seminiferous tubules with much reduced frequency. Lack of agreement with the RT-nested PCR assay results in testis tissues obtained at days 14, 28, and 59 postinfection suggested that PRRSV infection in the testis may be extremely restricted, and may not necessarily constitute a major viral source in semen during extended periods of seminal shedding.

Venereal transmission of pseudorabies viruses indigenous to feral swine.

Between 1995 and 1998, we designed a series of studies in which we attempted to determine the main routes of transmission involved in the natural infection of pseudorabies virus (PRV) indigenous to free-ranging feral swine (Sus scrofa). Naturally infected feral sows transmitted the infection to uninfected feral boars, with which they had been commingled for a 6-wk period. Pseudorabies virus was isolated from boar preputial swabs, but not from nasal swabs. Three of the same PRV-infected feral sows did not transmit the infection to domestic boars during a 16 wk commingling period, despite the fact that they became pregnant. Feral boars, naturally infected with PRV transmitted the virus to domestic gilts while penned together during 6 wk. Pseudorabies virus was isolated from vaginal swabs, but not from nasal swabs of gilts, after 2 and 3 wk of commingling. When the same infected boars were commingled with either feral or domestic boars for 13 wk, PRV transmission did not occur. None of the exposed boars developed neutralizing antibodies or yielded virus from their preputial or nasal swabs. Our results indicate that PRV indigenous to feral swine is preferentially transmitted to feral or domestic swine of the opposite sex by the venereal route. This mode of transmission differs from that seen in the natural transmission of PRV prevalent in domestic swine, where contaminated secretions, excretions and aerosols are responsible for the spread of the virus. Based on these results, we feel that as long as feral swine do not come into direct contact with domestic swine, PRV-infected feral swine probably pose only a limited risk to the success of the National Pseudorabies Eradication Program. The fact that PRV is usually transmitted from feral to domestic swine at the time of mating would indicate that the isolation of domestic herds by the use of a "double fence," should be adequate protection against reinfection with PRV.

Health status of bulls used for natural breeding on farms in south west Scotland.

One hundred and nine breeding bulls were examined during the period November 1992 to June 1993 on farms in south west Scotland for evidence of infectious diseases associated with breeding. Preputial washings were collected to screen for Campylobacter fetus venerialis, together with serial blood samples to assess their seroprevalence to Bovine Virus Diarrhoea virus (BVDv), Bovine Herpes Virus-1 (BHV-1), Leptospira hardjo and Bovine Herpes Virus-4 (BHV-4). The possible impact of natural mating on the epidemiology of these diseases is described. Evidence of infections with Campylobacter fetus and BVH-4 were not found in this sample. The overall seroprevalence to BVDv was 78%, for BHV-1 49%, and L. hardjo 27% at titres of > or = 1/400. This study shows that bulls may be responsible for the introduction and dissemination of these diseases when moved from farm to farm as part of normal cattle breeding in this area. Young unproven bulls may be particularly susceptible to endemic diseases associated with lowered reproductive performance.

Passive transfer, rate of decay, and protein specificity of antibodies against equine arteritis virus in horses from a Standardbred herd with high seroprevalence.

OBJECTIVE: To determine rate of decay of passively acquired antibodies in Standardbred foals on a farm with a high seroprevalence to equine arteritis virus (EAV) and to determine whether vertical or horizontal transmission of the virus was responsible for infection on the farm. DESIGN: Repeated-measures study. ANIMALS: 46 Standardbred horses (15 brood mares and their foals, 5 stallions, and 11 young horses). PROCEDURE: Serum samples obtained from horses on the farm were evaluated by serum neutralization and western immunoblot analysis to detect EAV-specific antibodies. The half-life of passively acquired antibodies in foals was estimated by use of regression analysis. RESULTS: Most (14/15) of the mares evaluated were seropositive to EAV. After suckling, their foals were also seropositive. Mean biological half-life for passively acquired antibodies in serum samples obtained from foals was 32 days (r2 = 0.61). The foal born to a seronegative dam and all 11 young horses from the farm were seronegative to EAV. At least 2 of 5 stallions on the farm were persistently infected carriers that were shedding virus in their semen. Immunoblot analysis of seropositive serum samples most consistently recognized the M protein of EAV. CLINICAL IMPLICATIONS: Analysis of these data indicated that a modified-live EAV vaccine can be administered to foals after they are 8 months old without risk of interference from maternal antibodies, regardless of serologic status of the foal's dam. Horizontal transmission of EAV via the respiratory tract apparently was uncommon on the farm, indicating that mares primarily were infected by venereal transmission of virus from carrier stallions.

Isolation of Coxiella burnetii from bull semen.

Previous observations on the dissemination of Coxiella burnetii between laboratory animals strongly support the hypothesis of venereal transmission. Serum and semen samples, from 57 bulls used for artificial insemination, were assayed for specific C. burnetii phase II antibodies and the presence of the organism respectively. Viable C. burnetii were detected in the semen of seropositive bulls. These findings indicate the possibility of sexual transmission of C. burnetii between cattle and further our knowledge of the epidemiology of the organism. The procedures used for investigations into the source of infection and route of tran-mission should be modified to take these findings into account.