A rare clinical presentation of human Dirofilaria repens infection as a pseudo-tumour of the epididymis - Case Report.

Introduction. Epididymal dirofilariasis is one of the unusual manifestations of this zoonosis. In Slovakia, this is the third case out of 20 Dirofilaria repens infected patients in whom the worm (the parasite) was identified in the epididymis. The patient felt a painless tumour about 2 cm in size on the left testicle. During ultrasound examination, the radiologist observed a cyst containing a live worm within the epididymis. After mechanical stimulation, the movement characteristic for filarial worms (´filarial dance sign´) appeared. An orchiectomy was performed at the Department of Urology in the University Hospital in Kosice. Histopathology confirmed a parasitic cyst with a worm, and based on characteristic morphological features, the parasite was identified as Dirofilaria repens. Objective. The aim of case report is to alert physicians to the possibility dirofilarial infection of the epididymis, where tumors and cystic structures often occur and to present clinical signs of the disease.

Reproductive injury in male BALB/c mice infected with Neospora caninum.

BACKGROUND: Neospora caninum is one of the main causes of abortion in pregnant animals. However, N. caninum-induced reproductive injury in male mice is still unclear. METHODS: Male BALB/c mice were infected with a bovine isolate of N. caninum, and the organ coefficients of the testis and epididymis were measured. Lesions in the testis and epididymis were observed by light microscopy and transmission electron microscopy. Expression of the spermatogenic cell apoptosis-related proteins p53 and caspase-3 was detected by western blot. The expression of spermatogenesis-related genes in the testis was detected by reverse transcription-PCR. Sperm morphology and motility were observed. The levels of nitric oxide (NO) and antisperm antibody (AsAb) in the testicular homogenates and hormones in the serum were detected by enzyme-linked immunosorbent assay. The reproductive capacity of the male mice was detected using a reproduction test. RESULTS: The organ coefficients of the testis and epididymis of the experimental group were significantly downregulated. Light microscopy examination revealed that the spermatogenic cells of the testis were arranged in a disordered manner, and the number was reduced. The number of sperm in the epididymal lumen was significantly reduced, and the cytoplasm exhibited vacuolation and degeneration. Ultrastructural studies revealed that the cells of the testis and epididymis tissues showed varying degrees of disease. The level of p53 and caspase-3 expression in the testis was significantly upregulated. The expression of the testicular spermatogenesis-related genes Herc4, Ipo11 and Mrto4 were strongly downregulated. Observation of sperm by microscopic examination revealed significantly reduced sperm density and sperm motility, and the number of sperm deformities was significantly increased. The level of NO and AsAb was significantly increased. The levels of luteinizing hormone, follicle-stimulating hormone and gonadotropin-releasing hormone were significantly upregulated, whereas the levels of testosterone, thyrotropin-releasing hormone, thyroxine and thyroid-stimulating hormone were significantly downregulated. After challenge, the infected male mice and healthy female mice were caged together: the subsequent fetal death rate was increased, and the conception rate, litter size, number of live births and the birth weight were significantly reduced. CONCLUSIONS: Infection of male BALB/c mice with the bovine isolate of N. caninum induced varying degrees of injury to the testis, epididymis and sperm of the mice, destroyed spermatogenesis and affected the reproductive capacity.

Pseudo-Tumor of the Epididymis, a Rare Clinical Presentation of Human Dirofilaria repens Infection: a Report of Autochthonous Case of Dirofilariasis in Southwestern Slovakia.

PURPOSE: Dirofilariasis caused by the filarial nematode Dirofilaria repens is mainly a disease of dogs and other carnivores. Also, humans can be accidentally infected with this parasite. The infective third-stage filariform larvae are transmitted by various species of mosquitoes. Until this day, a total of 17 human cases caused by D. repens have been diagnosed in Slovakia, 11 subcutaneous, 4 ocular, 1 pulmonary and 1 in the epididymis. The aim of this report was to describe an unusual clinical case of dirofilariasis of the scrotum. METHODS: Extirpated worm was subjected to the molecular and histological identification. PCR for the amplification of cytochrome oxidase subunit 1 (CO1) was performed using specific D. repens primer pair. RESULTS: Here we document the 13th case of human dirofilariasis in a 46-year-old man from southwestern Slovakia. Very rare in humans, genital involvement manifests itself as pseudotumor nodule affecting the epididymis. The patient consulted a general practitioner due to a palpable subcutaneous lump in the scrotum. Routine laboratory analysis revealed blood eosinophilia (16.6%). The ultrasound examination was indicated, and subsequently, surgical excision of the right epididymal nodule was performed. On the basis of histological microscopic examination and PCR-based detection, the helminth was identified as Dirofilaria repens. This represents the ninth case of autochthonous dirofilariasis in Slovakia. CONCLUSIONS: The majority of D. repens infections were recorded in southwestern regions of Slovak Republic, which are considered to be endemic areas for canine dirofilariasis. Our described patient also comes from southwestern part of Slovakia (Topolníky, Dunajská Streda region).

Trypanosoma brucei triggers a marked immune response in male reproductive organs.

African trypanosomiasis is caused by the protozoan parasite Trypanosoma brucei, transmitted between mammals by the bite of a tsetse. It has been recently shown that parasites accumulate in large numbers in various organs and tissues, including the mouse testis. Whether parasites are protected from the immune system in the male reproductive organ or can be transmitted through sexual route remains unknown. Here we show that parasites can be detected by fine needle aspiration cytology of the male reproductive system in mice, and histopathological analysis revealed that T. brucei accumulates in the stroma of the epididymis, epididymal adipose tissue and fibrous tunics of the testis. No parasites were found in the lumen of intact epididymal ducts or seminiferous tubules of the testis, indicating that the large majority of the parasites are not located in immune-privileged sites. In fact, these parasites are associated with marked inflammatory cell infiltration, parasite degeneration, and severe tissue damage and rupture of epididymal ducts, which may be related with reduced fertility. Overall, we show that just like in the bloodstream and most other tissues, in the male reproductive organs, T. brucei are exposed to a strong immune response. The detection of a very high number of parasites in this organ and its accessibility opens the possibility of using fine needle aspiration cytology as a complementary diagnostic tool in Animal African Trypanosomiasis.

Investigation of Toxoplasma gondii in semen, testicle and epididymis tissues of primo-infected cats (Felis catus).

This study aimed to investigate the presence of Toxoplasma gondii in semen, testicle and epididymis tissues of cats experimentally infected by this coccidium. A total of 12 male felines without a definite breed that were of reproductive age and serologically negative for T. gondii were selected and distributed to the following three experimental groups: GI, inoculated with 600 tissue cysts of the P strain of T. gondii (isolate III); GII, inoculated with 2×105 tachyzoites of the RH strain (isolate I); and GIII, not inoculated (control group). Prior to inoculation (day -7 and 0) and on post inoculation days (PIDs) 7, 14, 21, 28, 42, 56, and 70, all felines were subjected to assessments of anti-T. gondii IgG by indirect immunofluorescence (IIF) and assessments of parasitemia. Collection of semen (electroejaculation) was performed on the specified dates, followed by nested PCR and bioassays in mice to detect T. gondii. On PID 70, all 12 felines were orchiectomized, and the presence of the parasite in the testicles and epididymides was evaluated by nested PCR, murine bioassay, and histopathological and immunohistochemical analyses. All felines inoculated with T. gondii (GI and GII) seroconverted to the toxoplasmic infection after PID 14; on PID 7, seroconversion of three felines (P4, RH2 and RH4) could observed, and all exhibited detectable titers by PID 64. The GII felines exhibited greater serological titers compared with GI felines. The maximum serological titer (IgG) was observed in feline RH3 (titer 1024), while in other experimental felines, a maximum titer of 256 was detected. Parasitemic peaks were diagnosed in all felines of groups I and II from PIDs 7-42. A total of five parasitemic peaks were diagnosed in GI and nine in GII. In none of the experimental time points was the presence of T. gondii diagnosed in seminal samples collected from the felines or in the testicle or epididymis tissues collected from these animals. Thus, sexual transmission in domestic cats does not appear to be a major route of T. gondii infection, possibly demonstrating the tendency of this protozoan to develop a response directed to the formation and excretion of oocysts in the feces of these definite hosts, which act as its main route of perpetuation in the environment.

Potential sexual transmission of Trypanosoma cruzi in mice.

Infection with the protozoan parasite Trypanosoma cruzi, the etiologic agent of human Chagas disease, results in life-long infection. Infective trypomastigotes circulate in the bloodstream and have the capacity to infect any cell type, including reproductive tissue. This study sought to assess the potential for sexual transmission of T. cruzi in an experimental mouse model. The conditions used in this study, in which acutely infected males and immunosuppressing the females, created a worst-case scenario allowing for the greatest chance of measuring transmission through intercourse. Male BALB/c mice were infected and mated with uninfected females, and the females were subsequently examined for T. cruzi tissue parasitism. A single transmission event of 61 total matings was observed, indicating a low but non-zero risk potential for male-to-female sexual transmission of T. cruzi.

Toxoplasma gondii decreases the reproductive fitness in mice.

Toxoplasma gondii is a common protozoan parasite that infects warm-blooded animals throughout the world, including mice and humans. During infection, both, the parasite and the host, utilize various mechanisms to maximize their own reproductive success. Mice and humans are both the intermediate hosts for Toxoplasma gondii, which forms specialized vacuoles containing reproductive cysts in the formers' tissue. As half of the human population is infected, developing a disease called toxoplasmosis, along with an ever-growing number of couples suffering with idiopathic infertility, it is therefore surprising that there is a lack of research on how Toxoplasma gondii can alter reproductive parameters. In this study, a detailed histometric screening of the testicular function along with the levels of the pituitary luteinizing hormone (LH) were analysed in infected mice. Data on relative testis and epididymis weight, and sperm count were also collected. Based on the results obtained, the level of LH in the urine of Toxoplasma gondii infected mice was lower compared to the control. In direct correlation with the hormone level, testicular function and sperm production was also significantly lower in Toxoplasma gondii positive group using sperm count and histometric analysis as a marker. Not only were the number of leptotene primary spermatocytes and spermatids lowered, but the number of Sertoli cells and the tubule diameter were elevated. In parallel, a pilot epigenetic study on global testicular methylation, and specific methylation of Crem, Creb1 and Hspa1genes essential for successfully ongoing spermatogenesis was performed. Global methylation was elevated in Toxoplasma infected mice, and differences in the DNA methylation of selected genes were detected between the Toxoplasma positive and control group. These findings demonstrate a direct relation between Toxoplasma gondii infection and the decrease of male reproductive fitness in mice, which may contribute to an increase of idiopathic infertility in humans.

The "filarial dance" is not characteristic of filariasis: observations of "dancing megasperm" on high-resolution sonography in patients from nonendemic areas mimicking the filarial dance and a proposed mechanism for this phenomenon.

The objective of this series was to show that the sonographic appearance described as the "filarial dance" is not characteristic of filariasis but occurs in nonendemic areas as a manifestation of epididymal obstruction. An experienced observer documented cases after initial observation of the filarial dance in routine clinical practice using high-frequency linear array transducers. The filarial dance was described as excessive to-and-fro movement of echogenic particles within a prominent epididymis and graded 1 to 4 according to the extent and distribution of the abnormality. The country of birth, exposure to filarial infection or travel to a filarial-endemic area, previous scrotal surgery including vasectomy, any previous or current scrotal inflammatory disease, and any congenital testicular abnormalities were recorded. Over a 10-year period, sonographic appearances consistent with the filarial dance were observed in 18 patients (bilateral in 6). The mean patient age was 47.7 (range, 28-91) years. The abnormality was graded in the 24 affected testes as follows: grade 1, n = 3; grade 2, n = 8; grade 3, n = 8; and grade 4, n = 5. No patient had a history of filariasis or travel to an endemic area. Six of 18 patients (33.3%) had bilateral vasectomies; 5 (27.8%) had a history of epididymo-orchitis in the ipsilateral testis; 3 (16.7%) had previous scrotal surgery; and 4 (22.2%) had no relevant urologic history. We have described a sonographic appearance identical to the filarial dance in men with no history of filarial infection. Most had previous scrotal surgery or infection, suggesting that the filarial dance may not always be due to movement of filarial worms. The unifying condition in patients with filariasis and our patients is lymphatic obstruction, likely the underlying cause of the appearance in both groups.

Filariasis: an emergent cause of acute scrotal pain.

A 25-year-old man from Bangla Desh with acute right scrotal pain was subjected to scrotal surgical exploration because of the suspicion of testicular torsion. The testicle appeared normally positioned; an epididymal nodule was removed, and pathology showed the presence of the filaria worm. Filariasis is a tropical disease which has been estimated to affect 120 millions people throughout the world. Lymphadenitis and lymphangitis are the more common clinical settings; in men, there is a frequent scrotal involvement. In some cases, acute scrotal pain may lead to the suspicion of testicular torsion. The observation of patients with genital filariasis is likely to become more frequent in an era of massive immigration from different countries; nowadays, the disease should always be taken into consideration in the differential diagnosis in patients with acute scrotal pain coming from tropical areas.

Genital lesions associated with visceral leishmaniasis and shedding of Leishmania sp. in the semen of naturally infected dogs.

Although visceral leishmaniasis is primarily transmitted by a biological invertebrate vector, transmission in the absence of the vector has been reported, including venereal transmission in humans. Considering the possibility of venereal transmission, we studied genital lesions in dogs naturally infected with visceral leishmaniasis and shedding of Leishmania sp. in the semen. Approximately 200 dogs were serologically tested for anti-Leishmania antibodies and divided into three groups: 1) serologically negative dogs (n = 20), 2) asymptomatic serologically positive dogs (n = 20), and 3) symptomatic serologically positive dogs (n = 20). Samples from both testes, all segments of both epididymes, prostate gland, glans penis, and prepuce were histologically evaluated and processed for immunodetection of Leishmania sp. Semen samples were obtained from 22 symptomatic serologically positive dogs and processed for detecting Leishmania DNA by polymerase chain reaction. A significantly higher frequency of inflammation was observed in the epididymes, glans penis, and prepuce of dogs with visceral leishmaniasis, which was associated with a high frequency of immunohistochemically positive tissues (up to 95% of tissues from symptomatic dogs were positive by immunohistochemistry). Leishmania DNA was detected in eight of 22 semen samples from symptomatic dogs. Together these findings indicate that genital lesions and shedding of Leishmania sp. (donovani complex) in the semen are associated with visceral leishmaniasis. Additional studies should address the possibility of venereal transmission of the disease in the dog.

The occurrence of strongylid nematodes in the epididymides of wood mice.

The recent discovery of a larval nematode in the epididymides of free-living wood mice (Apodemus sylvaticus) suggests a sexual transmission of these parasites. They have been placed within the bursate nematodes (order Strongylida) through 18S rDNA analysis, suggesting that they are undetermined metastrongyloid nematodes. The possibility that these parasites are transmitted sexually opens an intriguing field of research because sexually transmitted metazoan parasites are known to occur mainly in invertebrates, whereas in vertebrates sexually transmitted parasites are usually microparasites such as viruses, bacteria and protozoa.

Sexual transmission of a nematode parasite of Wood Mice (Apodemus sylvaticus)?

Sexual transmission occurs commonly in microparasites such as viruses and bacteria, but this is an unusual transmission route for macroparasites. Here we present evidence which suggests that a nematode parasite of Wood Mice (Apodemus sylvaticus) may be sexually transmitted and we have classified the nematode using molecular data. Wood Mice were collected annually in the course of work on their reproductive physiology. Larval nematodes were found in the epididymides of 19.6% of males. It seems likely that they would be transmitted to females at ejaculation. To identify these larval nematodes, which we were unable to do using morphological features, we sequenced the 18S rDNA. Sequence comparisons with the molecular phylogeny of Blaxter et al. (1998) demonstrated that they were bursate nematodes (Order Strongylida). The relationships between strongylid taxa were poorly resolved by 18S rDNA. However, both distance and parsimony analyses grouped the nematode with the superfamily Metastrongylidea in a clade containing Filaroides and Angiostrongylus sp. Importantly, the sequences were distinct from those of Heligmosomoides polygyrus and Angiostrongylus dujardini, two common strongylid nematodes of Apodemus. We were therefore unable positively to identify these worms by matching their sequences with those from morphologically identifiable adult strongylid nematodes infecting Apodemus. These results demonstrate that an as yet unidentified strongylid is quite commonly found in large numbers in the male reproductive tract of Wood Mice. Further work is required to understand the biology and transmission dynamics of this interesting system.

Genitourinary changes in hamsters infected and reinfected with Trypanosoma cruzi.

Authors describe genitourinary changes in male hamsters infected and reinfected with Trypanosoma cruzi. Changes in genital organs have been described in human and in experimental chagasic infection. Genital dysfunctions in chronic chagasic patients affect ejaculation, libido and sexual potency, and testis biopsies may show arrested maturation of germ cells, oligozoospermia and azoospermia. Sixty-five male hamsters were inoculated and reinoculated with 2x10 trypomastigotes of T. cruzi VIC strain, and 22 non-infected animals constituted the control group. Animals were necropsied and fragments from testis, epididymis, seminal vesicle and bladder were collected and stained with hematoxylin-eosin. Peroxidase anti-peroxidase procedure was utilized to detect tissue parasitism. T. cruzi nests were found in testis, epididymis and seminal vesicle of these hamsters. Such parasitism plays a role in the origin of genital lesions observed in humans and laboratory animals during chronic chagasic infection.

Microsurgical epididymal sperm aspiration with motile trophozoite cells but no spermatozoa.

This paper reports on a patient in whom the clinical diagnosis of obstructive azoospermia was made according to clinical observations, i.e. azoospermia, normal andrological examination, normal follicle stimulating hormone and a misleading histopathological report of a testicular biopsy. Microsurgical vasoepididymostomy failed to restore fertility, and as a last resort, microsurgical sperm aspiration was performed. Although flagellated cells were observed in the epididymal aspiration, no spermatozoa were observed and wet preparation of multiple testicular biopsies failed to demonstrate any spermatozoon. This patient was diagnosed to have a non-obstructive azoospermia, resulting from maturation arrest associated with trichomonas infection at the level of the epididymis.

An epizootic of besnoitiosis in goats in Fars province of Iran.

Over 100,000 goats were grazing freely in isolated herds of 100 to 600 goats in a mountainous area of more than 2000 km2 to the north-east of Darab City in Fars Province in southern Iran. Twenty to fifty percent of the goats in this area were infected with besnoitiosis. Sheep, cattle and donkeys grazing in the same area showed no clinical manifestations of the infection. The diagnosis of the infection in the clinically suspected animals was confirmed by histological studies on skin biopsies from the ear tips and carpal and tarsal regions, and the distribution of this protozoon in the tissues and organs of this intermediate host was studied by post-mortem and histological examinations. Skin biopsies from the carpal and tarsal areas of 12% of clinically normal goats from the infected area were also lightly infected with besnoitia cysts.

Besnoitiosis of the reproductive tract of a Blue Duiker (Cephalophus monticola).

A mature male Blue Duiker that had been born in the United States was submitted for necropsy examination following a brief illness. On histologic examination of the reproductive tract several Besnoitia cysts were found in the epididymis, prostate and bulbourethral gland. The lack of an inflammatory response or any negative effect on fertility, based on histologic evaluation and breeding history, is in contrast with the severe orchitis, epididymitis and infertility of besnoitiosis in cattle. This is the first report of an autochthonous infection of Besnoitia in the United States as well as the first report of besnoitiosis in a Blue Duiker.