Testicular pain as an unusual presentation of COVID-19: a brief review of SARS-CoV-2 and the testis.

RESEARCH QUESTION: Can the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) virus induce testis damage and dysfunction? DESIGN: This is the description of the case of a young man presenting with heavy testicular pain as the first symptom of COVID-19 infection. A review of the literature is also presented. RESULTS: SARS-CoV-2 may enter into the host cell by binding to angiotensin-converting enzyme 2. This receptor seems to be widely expressed in different testicular cell types, making possible the occurrence of orchitis in male patients with COVID-19 infection. From a review of the literature, it seems that there is currently no evidence of sexual transmission of SARS-CoV-2; however, the possibility of virus-induced testis damage and dysfunction cannot be excluded. CONCLUSIONS: Further studies are necessary on the pathological effect of SARS-CoV-2 in the male reproductive system and to ensure a proper andrological follow-up for male patients.

Newcastle disease virus induces testicular damage and disrupts steroidogenesis in specific pathogen free roosters.

Newcastle disease (ND), which is caused by Newcastle disease virus (NDV), can cause heavy economic losses to the poultry industry worldwide. It is characterised by extensive pathologies of the digestive, respiratory, and nervous systems and can cause severe damage to the reproductive system of egg-laying hens. However, it is unknown whether NDV replicates in the male reproductive system of chickens and induces any pathologies. In this study, we selected a representative strain (i.e. ZJ1) of the most common genotype (i.e. VII) of NDV to investigate whether NDV can induce histological, hormonal, and inflammatory responses in the testes of specific pathogen free (SPF) roosters. NDV infection increased the expression of toll like receptor TLR3, TLR7, MDA5, IFN-α, IFN-ß, IFN-γ, IL-8, and CXCLi1 in the testes of NDV-infected roosters at 5 days post-infection (dpi). Severe histological changes, including decrease in the number of Sertoli cells and individualized, shrunken spermatogonia with pyknotic nuclei, were observed at 3 dpi. At 5 dpi, the spermatogenic columns were disorganized, and there were fewer cells, which were replaced by necrotic cells, lipid vacuoles, and proteinaceous homogenous material. A significant decrease in the plasma concentrations of testosterone and luteinizing hormone (LH) and the mRNA expression of their receptors in the testes, steroidogenic acute regulatory protein, cytochrome P450 side-chain cleavage enzyme, and 3ß-hydroxysteroid dehydrogenase in the NDV-infected group was observed relative to those in the control group (P < 0.05). Collectively, these results indicate that NDV infection induces a severe inflammatory response and histological changes, which decrease the steroidogenesis.

Safety and efficacy of vaccination of seronegative bulls with modified-live, cytopathic bovine viral diarrhea viruses.

The objectives were to vaccinate peri-pubertal bulls with a modified-live vaccine consisting of cytopathic BVDV strains Singer and 296 and evaluate the resulting: (a) transient shed of modified-live, cytopathic BVDV in semen; (b) risk of prolonged testicular infection; and (c) protection against subsequent testicular infection due to viral challenge. Seronegative, peri-pubertal bulls were vaccinated subcutaneously with a standard dose of vaccine (n=11) or were maintained as unvaccinated controls (n=11). Forty-nine days after vaccination, all bulls were intranasally inoculated with a noncytopathic field strain of BVDV. Semen and testicular biopsies collected after vaccination and challenge were assayed for BVDV using virus isolation, reverse transcription-nested PCR, or immunohistochemistry, and the identity of viral strains was determined by nucleotide sequencing of PCR products. Vaccination of peri-pubertal bulls with this vaccine caused a short-term, transient shed of only the type 1a strain of modified-live, cytopathic BVDV in semen for up to 10d after vaccination. The vaccine did not cause prolonged testicular infection. Vaccination with this product prevented development of prolonged testicular infections after subsequent exposure to a field strain of BVDV.

Testicular hypoplasia in a bull persistently infected with bovine diarrhoea virus.

A bull aged 16 months with bilateral testicular hypoplasia and azoospermia was persistently infected with bovine viral diarrhoea virus (BVDV). Viral antigen was detected in serum and semen by ELISA, but the animal was serologically negative. After slaughter, the genital tract was examined histopathologically and by immunohistochemistry, including double immunolabelling with BVDV antibody and either S-100 antibody (for Sertoli cells) or ferritin antibody (for Leydig cells). The seminiferous tubules of both testes were lined by a single layer of Sertoli cells and the germinal epithelium was completely absent except for a few remaining spermatogonia. BVDV antigen was demonstrated (1) in the media of arterial vessel walls of the testis, epididymis, urethra, prostate, and vesicular and bulbourethral glands, (2) in epithelial cells of the ductus epididymidis, the accessory glands and the urethra, and (3) in the testis, mainly in Sertoli cells and to a lesser extent in the spermatogonia that remained, but not in Leydig cells. The testicular hypoplasia was possibly linked to the BVDV infection.

Semen alterations in porcine rubulavirus-infected boars are related to viral excretion and have implications for artificial insemination.

Porcine rubulavirus (PoRV), also known as blue eye disease (BED) of swine, causes respiratory and reproductive problems in pigs at several developmental stages. To study the effect of PoRV infection on semen production, five boars were infected with 1 x 10(6) TCID(50)/ml of PoRV strain PAC-3 and evaluated for 59 days post inoculation (DPI). Infected boars developed reproductive tract pathology that included swelling of the testes and epididymides. Analysis of the semen showed that the infection had little effect on semen production in four animals, but semen from one boar showed severe alterations in sperm concentration, motility, and morphology. When motility was analyzed in BTS-diluted semen after 24, 48, or 72 h, alterations were detected in all boars. Furthermore, viral antigen was detected in semen, the seminal plasma fraction, or sperm fraction from all boars. These results showed that PoRV is excreted via semen and, therefore, artificial insemination is a potential route of dissemination.

Concomitant testicular infection by Zika virus and Schistosoma mansoni in a Brazilian young boy.

Sumary The identification of a escrotal mass without pain or report of trauma should be investigated to rule out scrotal cancer. We report the case of a young Brazilian boy who underwent orchiectomy after magnetic resonance imaging (MRI) and duplex scan (DS) indicating a high possibility of cancer. Blood exams ruled out the possibility of cancer. Testicular biopsy was not indicated. After surgery the diagnostic was extensive orchiepididymitis by Schistosoma. In endemic areas orchiepididymis by Schistosoma should be investigate to avoid unnecessary surgeries. This patient was also infected with Zika virus.

Use of a modified-live vaccine to prevent persistent testicular infection with bovine viral diarrhea virus.

A commercial vaccine containing modified-live bovine viral diarrhea virus (BVDV; types 1 and 2) was administered to one group of 22 peripubertal bulls 28 days before intranasal inoculation with a type 1 strain of BVDV. A second group of 23 peripubertal bulls did not receive the modified-live BVDV vaccine before intranasal inoculation. Ten of 23 unvaccinated bulls--but none of the vaccinated bulls--developed a persistent testicular infection as determined by immunohistochemistry and polymerase chain reaction. Results of this study indicate that administration of a modified-live vaccine containing BVDV can prevent persistent testicular infection if peripubertal bulls are vaccinated before viral exposure.

[Experimental study on effects of testis murine cytomegalovirus infection on sperm viability in mice].

OBJECTIVE: To explore the effects of testis murine cytomegalovirus (MCMV) infection on mature sperm viability in mice. METHODS: BALB/c mice without MCMV infection, screened by ELISA, were randomly divided into two groups: an experimental group (n = 64) and a control group (n = 40). The former were directly inoculated with MCMV into the testis, while the latter treated by inoculation of DMEM without MCMV. The mice in both of the groups were sacrificed respectively at 1, 2, 4, 6, 9, 14, 21, 38 d postinoculation (D1, 2, 4, 6, 9, 14, 21, 38 PI), the testis was examined histopathologically, and meanwhile the viability of mature sperms in the epididymis cauda was measured. RESULTS: MCMV basophil inclusion bodies were found in the Leydig cells in the experimental group, and spermatogenic cells were vacuolated and arranged disorderly. Compared with the control group, the sperm viability in the experimental group was decreased significantly by 71.42% to 56.04% (P < 0.05) on D1 PI. CONCLUSION: The sperm viability in mice might be descended significantly by MCMV infection in the early period, but restored to normal with time. This shows that MCMV infection might influence procreation transiently.

Testicular lesions and antler abnormalities in Colorado, USA mule deer (Odocoileus hemionus): a possible role for epizootic hemorrhagic disease virus.

Antler abnormalities of deer and other cervids often result from testicular lesions and decreased levels of testosterone, inhibiting normal cycles of antler growth. Affected males have antlers with retained velvet, numerous short, misshapen points ("cactus bucks"), and failure to shed these abnormal antlers annually. In Colorado, US, we observed a high occurrence of "cactus bucks" in mule deer (Odocoileus hemionus) populations after management efforts to increase the number of mature male deer in the state. Affected males consistently had antibody to epizootic hemorrhagic disease virus serotype 2 (EHDV-2), and examination of the testes of these animals demonstrated nonspecific end-stage lesions of chronic inflammation, fibrosis, and mineralization. To examine more acute stages of testicular lesions, and to screen for EHDV specifically within the testes, we sampled 16 male mule deer from affected herds, but with essentially normal antlers (n = 14) or retained velvet only (n = 2). Testicular and epididymal lesions identified from these samples included necrotizing vasculitis (n = 2), hemorrhage (n = 6), edema (n = 2), seminiferous tubular necrosis (n = 5), orchitis (n = 5), epididymitis (n = 10), hypospermia (n = 6), and end-stage lesions of seminiferous tubular loss (n = 2), fibrosis (n = 2), and mineralization (n = 2). Each of the 16 cases was blindly scored on the basis of number of histologic lesions, with a median score of two. Five of seven (71%) testes that were PCR positive for EHDV had lesion scores above the median, whereas none of the nine (0%) EHDV PCR-negative testes had lesion scores above the median, suggesting an association between testicular lesions and detection of EHDV RNA in the testes (P = 0.003). Although the role of EHDV infection remains unconfirmed, the association between testicular and epididymal lesions and presence of EHDV RNA in the affected tissues suggests that cactus buck antlers may be a sequela of EHDV infection.

Persistent bovine pestivirus infection localized in the testes of an immuno-competent, non-viraemic bull.

A post-pubertal bull on an artificial insemination station was found to be persistently shedding bovine viral diarrhoea virus (BVDV) in semen over a period of eleven months, while demonstrating no viraemia. Circulating antibodies to BVDV were consistently high, suggesting that the immune system was challenged repeatedly. Post-mortem findings confirmed that the virus was sequestered in the testes of the bull. It is hypothesized that the BVDV in this immuno-competent bull was protected from the bull's immune response by the blood-testes barrier. The barrier becomes functional only at puberty when tight junctions form between adjacent Sertoli cells, suggesting that this bull became persistently infected with BVDV during puberty.

Avian infectious bronchitis virus: a possible cause of reduced fertility in the rooster.

The formation of epididymal stones in the rooster epididymis is a widespread problem that has detrimental effects on sperm production and fertility. The cause of epididymal stones is unknown, but an infectious agent, the avian infectious bronchitis virus (AIBV), has been implicated. The goal of this study was to determine if administering the live attenuated AIBV vaccine to male chicks increases the incidence of stones in the epididymal region of the adult rooster. Specific pathogen free (SPF) Leghorn roosters were divided into two groups: a vaccine-free group (n = 7) and a group vaccinated with AIBV (n = 12). The vaccine was administered orally at 2, 4, 10, and 14 wk of age. Blood was drawn weekly to monitor antibodies to AIBV. At 26 wk of age, blood was obtained to determine testosterone concentrations, and reproductive tracts were removed to analyze daily sperm production and to detect epididymal stones. Nine of 12 vaccinated roosters developed stones, whereas those not given the vaccine did not develop stones. Serum testosterone concentrations were significantly (P < 0.05) reduced in vaccinated roosters with epididymal stones (3.6 +/- 0.30 ng/ml) when compared with nonvaccinated roosters that did not have epididymal stones (7.0 +/- 1.63 ng/ml). Testis weight was significantly (P < 0.05) reduced in vaccinated roosters with epididymal stones (12.1 +/- 0.76 g), as compared with nonvaccinated roosters without epididymal stones (15.2 +/- 0.81 g). Daily sperm production was significantly (P < 0.05) decreased in vaccinated roosters with epididymal stones (5.03 +/- 0.31 x 10(8) sperm/testis/day) when compared with nonvaccinated roosters without epididymal stones (7.43 +/- 0.52 x 10(8) sperm/testis/day). Comparing daily sperm production on a per gram basis, vaccinated roosters with epididymal stones had 4.38 +/- 0.14 x 10(7) sperm/g of testis, which was significantly (P < 0.05) smaller than nonvaccinated roosters without epididymal stones, which had 5.17 +/- 0.17 x 10(7) sperm/g of testis. We conclude that the use of a live attenuated AIBV vaccine increases the incidence of epididymal stones in roosters, resulting in decreased sperm production and decreased serum testosterone concentrations.

Lesions in the reproductive tract of boars experimentally infected with porcine rubulavirus.

"Blue eye" disease of pigs in Mexico is caused by porcine rubulavirus and characterized by infertility in sows and boars, nervous signs in young pigs, and corneal opacity in pigs of all ages. The pathogenesis of reproductive tract lesions in rubulavirus-infected boars has not previously been investigated. In a first experiment, four 9-month-old boars were inoculated with porcine rubulavirus and killed 5, 15, 30 or 45 days post-inoculation (pi). In a second experiment, four similar boars were inoculated with the same virus and two animals were killed on each of days 70 and 80 pi. Swelling of the head of the epididymis developed in all inoculated boars at approximately day 15 pi. Reduced spermatozoan motility and concentration were detected in semen samples collected from one boar from day 21 pi. At post-mortem examination, nodules were seen in the head of the epididymis of the boars killed 15, 30 or 45 days pi and the right testis of the pig killed 30 days pi was atrophic. Corresponding histopathological epididymal alterations included formation of spermatic granulomas and vacuolar degeneration of ductular epithelium. These lesions were associated with mononuclear cell infiltration and interstitial fibroplasia. Degeneration of seminiferous tubules and interstitial mononuclear cell infiltration were seen in the atrophic testis of the pig killed 30 days pi. There was fibrosis of the head of the epididymis in all boars killed 70 or 80 days pi and one of these animals also had right testicular atrophy associated with degeneration of seminiferous tubules, lymphocytic infiltration and giant cell formation. Porcine rubulavirus antigen was detected by immunofluorescence labelling in the head of the epididymis of the pigs killed 15, 30 or 45 days pi and in one animal killed on day 70 pi. These results indicate that porcine rubulavirus can cause severe epididymo-orchitis and reduced semen quality in sexually mature boars.

Encephalomyocarditis (EMC) virus-induced testicular lesion in BALB/c mice.

Characteristics of encephalomyocarditis (EMC) virus-induced testicular lesions were investigated in 4- and 8-week-old BALB/c male mice after intraperitoneal (i.p.) and intratesticular (left) (i.t.) inoculation of the D variant of EMC virus (EMC-D). Apart from variation in severity and incidence, the histopathological nature of the resultant testicular lesion was similar in all infected mice, and was characterized by degeneration and necrosis of germinal cells and spermatogonia with inflammatory infiltration. Almost all the inoculated left testes of the i.t. group developed marked lesions. In general, the virus titre in the testis and incidence of testicular lesions were higher in 4-week-old mice than in 8-week-old mice. In addition, testicular lesions developed earlier and with a higher incidence in the PBS-inoculated right testis of the i.t. group than in either testis of the i.p. group of the same age.

Laboratory diagnosis and transmissibility of bovine viral diarrhea virus from a bull with a persistent testicular infection.

Recently, in the United States, a dairy bull was diagnosed as the second confirmed case of persistent testicular infection (PTI) with bovine viral diarrhea virus (BVDV). The first objective of this study was to evaluate the testing methodologies currently used by the artificial insemination industry in order to improve the detection of bulls with PTI. This study evaluated the impact of multiple factors ([1] sample tested, [2] sample handling, [3] assay used, and [4] assay methodology) on the sensitivity of detection of BVDV. The second objective of this study was to evaluate the transmissibility of BVDV from the bull through casual or sexual contact. Results from this study indicate that straws of semen should be transported to the diagnostic laboratory in liquid nitrogen dry shippers. PCR proved to be a more sensitive assay than virus isolation; however, certain PCR protocols exhibited greater diagnostic sensitivity than others. Insemination with cryopreserved semen from this infected bull caused viral transmission to a seronegative heifer resulting in viremia and seroconversion. After 42 months of age, the bull appeared to clear the infection. In conclusion, this bull validates that natural exposure to a 1a strain of BVDV can result in a unique PTI causing contamination of semen with detectable infectious virus. Appropriate handling and testing of samples is necessary in order to detect bulls exhibiting PTI. Additionally, PTI with BVDV may potentially be cleared after an extended duration.

Concomitant testicular infection by Zika virus and Schistosoma mansoni in a Brazilian young boy / Infecção testicular associada com Zika vírus e Schistosoma mansoni em jovem brasileiro

Sumary The identification of a escrotal mass without pain or report of trauma should be investigated to rule out scrotal cancer. We report the case of a young Brazilian boy who underwent orchiectomy after magnetic resonance imaging (MRI) and duplex scan (DS) indicating a high possibility of cancer. Blood exams ruled out the possibility of cancer. Testicular biopsy was not indicated. After surgery the diagnostic was extensive orchiepididymitis by Schistosoma. In endemic areas orchiepididymis by Schistosoma should be investigate to avoid unnecessary surgeries. This patient was also infected with Zika virus.

Resumo A identificação de massa escrotal sem relato de dor ou trauma deve ser investigada a fim de afastar a possibilidade de câncer escrotal. O artigo reporta o caso de um jovem brasileiro que apresentou massa escrotal com essas características. Ressonância nuclear magnética (RNM) e ultrassonografia (US) indicaram grande possibilidade de câncer. Os marcadores tumorais sanguíneos estavam normais, e a biópsia não poderia ser realizada. O resultado anatomopatológico diagnosticou extensa fibrose esquistossomótica, associada a quadro clínico e sorológico de Zika vírus concomitantemente. Em regiões endêmicas, pacientes com alterações escrotais devem ser pesquisados a fim de evitar cirurgias desnecessárias.

Epidemiology of prolonged testicular infections with bovine viral diarrhea virus.

Previously, bovine viral diarrhea virus (BVDV) had been found in prolonged testicular infections following acute infection of immunocompetent bulls. The primary purpose of this research was to evaluate the production and maintenance of prolonged testicular infections after exposure to BVDV of seronegative bulls in varying circumstances. The secondary objective was to initiate assessment of the potential for transmission of BVDV via semen of bulls exhibiting a prolonged testicular infection. In total, 10 research trials were conducted. The first trial examined the duration of detectable virus in semen after intranasal inoculation of peri-pubertal bulls. The second to fifth trials examined the potential for prolonged testicular infections resulting from natural exposure of seronegative bulls to persistently infected heifers. In the last five trials, the potential for viral transmission from bulls exhibiting prolonged testicular infections to a small number of exposed animals (n=28) was evaluated. Results of this research demonstrated that prolonged testicular infections could result in detection of viral RNA in semen for 2.75 years with infectious virus grown from testicular tissue 12.5 months after viral exposure. A type 1b strain of BVDV caused prolonged testicular infection after natural exposure of seronegative bulls to a persistently infected heifer. However, transmission of BVDV to susceptible animals was not detected in the final five trials of this research. In conclusion, BVDV can persist in testicular tissue after acute infection for several years, but the potential for viral transmission from these prolonged testicular infections appears to be low.

Cyclophosphamide-immunodepressed FVB/N mice: potentiating the effects of testicular cytomegalovirus infection.

The aim of this study was to investigate the effect of cyclophosphamide (CY) immunosuppression on the infection of germinal cells following testicular inoculation of male FVB/N mice with murine cytomegalovirus (MCMV). We used CY to modulate the immune status in order to mimic iatrogenic immunosuppression in humans (organ transplantation) as closely as possible. We show that viral pathological manifestations observed in mice treated with this CY-MCMV combination were severer than those observed in immunocompetent male mice infected with MCMV alone. As previously reported, the typical MCMV cellular inclusions were present in interstitial spaces; however, the spermatogenic cells were never directly infected. Nonetheless, at the end of our observation, we obtained definitive necrosis of the testes. These results suggest that germline cell necrosis induces sterility in immunodepressed infected male mice indirectly. In the case of organ transplantation, particular attention should be accorded to male patients receiving CY.

Association of human papillomavirus type 6 with a verruciform xanthoma.

We report a case of verruciform xanthoma (VX) associated with human papillomavirus (HPV) in a 67-year-old male. The patient had a pale-reddish, granular and verrucous tumor on the right side of his scrotum for four years. Histopathologic examination showed typical features of VX. HPV was detected by immunohistochemistry, electron microscopy, and PCR examinations. Ultrastructural examination revealed virus-like particles of 40-50 nm in the nucleus of the upper epidermal keratinocytes. HPV type 6a DNA was detected in lesional tissue by polymerase chain reaction and sequence analysis. To the best of our knowledge, this is the first case report of VX associated with HPV.