Usual or unusual presentations of Dirofilaria repens in two sibling dogs: a case report.

This study describes two different manifestations of Dirofilaria repens infection in sibling dogs with microfilaremia. Dog 1, asymptomatic, harbored a gravid female of D. repens on the parietal layer of tunica vaginalis of one testicle and showed a marked circulating eosinophilia (3.3·103/µL). Both testicles were normal in shape and size without any gross lesions. Dog 2 had a pyotraumatic dermatitis. The cases were confirmed by PCR and sequencing. The sequences obtained showed 100% identity with those of D. repens isolated from human scrotum in Croatia. The treatment with moxidectin 2.5% and imidacloprid 10%/kg was effective in eliminating microfilariae after just one application, as demonstrated by negative modified Knott's tests and PCR analyses of blood samples. This status was maintained during the post-treatment observation period. The classical localization of D. repens in dogs is in subcutaneous tissues, within nodules or free; however, it can also occur with some frequency in testicles, as described in humans. The infection can be associated with circulating eosinophilia or pyotraumatic dermatitis, as reported in this study. Thus, in endemic areas, it is advisable to carefully inspect the removed testicles at neutering since parasite localization can take place without any macroscopic changes. Moreover, in the case of circulating eosinophilia or pyotraumatic dermatitis, investigations should include modified Knott's test and PCR to ensure that D. repens is not the cause of these alterations. Rapid and sensitive tests for the early detection of infected animals would help to prevent or limit the spread of this zoonosis.

Schistosomiasis (bilharziasis) and male infertility.

Schistosomiasis (bilharziasis) is an endemic parasitic disease of a major source of morbidity and mortality for developing countries. It is caused by different species of blood flukes (trematode worms) of the genus Schistosoma, the most important of which are S. haematobium which infests the genitourinary tract and S. mansoni and S. japonicum which infest the gastrointestinal tract. The incidence of schistosome-induced male infertility is not known and probably underestimated being overshadowed by the more severe and often fatal urinary and gastrointestinal complications. Several causes may contribute to schistosomiasis-induced male infertility, such as hormonal imbalance, testicular tissue damage and genital ductal system obstruction, in addition to decreased libido and erectile dysfunction due to associated hormonal imbalance and prostatic infestation. Demonstration of the schistosome ova in seminal plasma or testicular tissue confirms the diagnosis. Treatment of schistosomiasis-induced male infertility depends on clinical evaluation and includes, besides anthelmintic treatment, hormonal replacement and assisted reproduction (IVF/ICSI) in cases of severe oligozoospermia and or obstructive and nonobstructive azoospermia.

Evidence for Previously Unidentified Sexual Transmission of Protozoan Parasites.

Knowing the mode of transmission of a disease can affect its control and prevention. Here, we identify 5 protozoan parasites with demonstrated presence in seminal fluid, only 1 of which has been identified as a sexually transmitted disease among humans.

Schistosomiasis (Bilharziasis) ova: An incidental finding in testicular tissue of an obstructive azoospermic man.

Congenital bilateral absence of vas deferens (CBAVD) results in obstructive azoospermia in which testicular function, such as spermatogenesis, is preserved. Bilateral testicular biopsy is not only diagnostic but also therapeutic as retrieved spermatozoa are usually cryopreserved for assisted reproduction. In this case report, CBAVD was confirmed in a 24-year-old azoospermic man on the basis of persistent azoospermia associated with low semen volume, absent fructose and normal hormonal levels, ultrasonographically (absent seminal vesicles) and histologically (intact spermatogenesis). Interestingly and incidentally, only the right testicular biopsy showed ova of two parasitic species of Schistosoma, namely Schistosoma haematobium which infests the genitourinary tract and Schistosoma mansoni which infests the gastrointestinal tract. Both species are rare causes of azoospermia, and the case should be further managed as CBAVD in which the left testicle is considered the preferred site of sperm retrieval for assisted reproduction.

Morphological and ultrastructural characteristics of Myxobolus ridibundae n. sp. (Myxosporea: Bivalvulida) infecting the testicular tissue of the marsh frog Rana ridibunda (Amphibia: Ranidae) in Egypt.

Myxozoans are one of the most economically important groups of protozoan parasites causing many serious diseases of their hosts. In the present study, a total of 60 live adult male specimens of the marsh frog Rana ridibunda have been randomly captured during the period of January-December 2015 in different areas at Kafr El-Sheikh Governorate, Egypt and were examined for infection by myxosporidian parasites. A total of 48 (80.0 %) out of 60 frog specimens were found to be infected with Myxobolus species. Parasitic infection was restricted to the testicular tissue of the examined frogs. Macroscopic cysts (plasmodia) which heavily infested different parts of the testes were recovered. Morphological and ultrastructural characteristics of these myxosporidian species were carried out using light and transmission electron microscopy. Plasmodia measured 0.16-0.53 (0.34 ± 0.01) mm in diameter. Mature spores appeared oval in frontal view, measuring 8.9-11.5 (9.6 ± 0.1) µm in length and 7.5-9.1 (8.4 ± 0.1) µm in width containing 5-6 turns of polar filaments. Morphometric characterization revealed that the very small size of the present Myxobolus species was the most distinctive feature that separates them from all previously described Myxobolus species. Ultrastructural analysis showed that the plasmodia are surrounded by a plasma membrane with numerous pinocytotic protrusions extending toward the host cell. The generative cells and the different developmental stages are arranged at the periphery of the plasmodia, while immature and mature spores are centrally located. Sporogenesis, capsulogenesis, valvogenesis, and spore maturation of the present parasite are also described. The present species is described as Myxobolus ridibundae and represents a new species.

Parasitism and inflammation in ear skin and in genital tissues of symptomatic and asymptomatic male dogs with visceral leishmaniasis.

Canine visceral leishmaniasis (CVL) is transmitted through vector, although venereal transmission has been suggested. This study aimed to compare the parasitic loads and inflammatory processes in genital tissues with ear skin from seropositive male dogs. Forty-five seropositive dogs were separated into groups containing symptomatic (n = 23) and asymptomatic (n = 22) animals. The control group (n = 2) healthy animals with seronegative and negative results in direct parasitological test. Samples of ear tip skin, prepuce, glans penis, testis, epididymis, and prostate were collected for evaluation of parasitic load and inflammatory infiltrate. Although ear tip skin was the most intensely parasitized, prepuce and epididymis revealed no difference in parasitism when compared with ear tip skin (P > 0.05). Parasitic loads in testis and prostate were lower than other tissues (P < 0.05). Parasitism in glans penis was high, similar to prepuce and epididymis, but lower than ear tip skin. High parasitism was more frequent in symptomatic dogs than asymptomatic animals. Severe inflammatory processes were more frequent within the symptomatic animals compared with asymptomatic and more predominant in prepuce and epididymis. Ear tip skin and genital tissues presented signs of chronic inflammation. There were weak and moderate positive correlations between parasitic loads and inflammatory processes. Our results demonstrate that, likewise with the ear tip skin, the genital of seropositive dogs can carry a large number of Leishmania infantum amastigotes and this process are more intense in symptomatic animals. These data have important implications for understanding the possibility of venereal transmission of CVL.

Hymenolepis folkertsi n. sp. (Eucestoda: Hymenolepididae) in the oldfield mouse Peromyscus polionotus (Wagner) (Rodentia: Cricetidae: Neotominae) from the southeastern Nearctic with comments on tapeworm faunal diversity among deer mice.

A previously unrecognized species of hymenolepidid cestode attributable to Hymenolepis is described based on specimens in Peromyscus polionotus, oldfield mouse, from Georgia near the southeastern coast of continental North America. Specimens of Hymenolepis folkertsi n. sp. differ from those attributed to most other species in the genus by having testes arranged in a triangle and a scolex with a prominent rostrum-like protrusion. The newly recognized species is further distinguished by the relative position and length of the cirrus sac, shape of seminal receptacle, and relative size of external seminal vesicle and seminal receptacle. Hymenolepidid cestodes have sporadically been reported among the highly diverse assemblage of Peromyscus which includes 56 distinct species in the Nearctic. Although the host genus has a great temporal duration and is endemic to the Nearctic, current evidence suggests that tapeworm faunal diversity reflects relatively recent assembly through bouts of host switching among other cricetid, murid, and geomyid rodents in sympatry.

Relationship between testicular lesion and hormone levels in male rats infected with Trypanosoma evansi.

The aim of this study was to evaluate the relationship between testicular lesions and hormone levels in rats experimentally infected with Trypanosoma evansi. For that, the measurement of reproductive hormones, histopathology and biomarkers of cellular injury were carried out in twenty-four animals, which were divided into two groups with 12 animals each. Group A was the negative control, or uninfected, while group B was composed by animals infected with T. evansi. Both groups were divided again into two other subgroups (n=6), from which serum and testicular fragments were collected on days 5 (A1 and B1) and 15 (A2 and B2) post-infection (PI). The morphological analysis showed increased alterations of head and tail of sperm in infected rats when compared with those of the control group. A significant reduction (P<0.01) in the levels of LH, FSH, testosterone and estradiol, associated with an increase in cortisol, was observed in serum of group B when compared with negative control. Additionally, NOx, lipid peroxidation and protein oxidation were enhanced in testicles, indicating the occurrence of cellular lesion. On histopathology, it was possible to observe testicular degeneration, among other disorders in infected animals. Therefore, based on these results, it is possible to conclude that the experimental infection with T. evansi caused changes in the levels of the main hormones of male rats associated with cellular injury.

Three new gonad-infecting species of Philometra (Nematoda: Philometridae) parasitic in Lutjanus spp. (Lutjanidae) in the northern Gulf of Mexico off Florida, USA.

Based on light and scanning electron microscopical studies, three new gonad-infecting species of Philometra Costa, 1845 (Nematoda: Philometridae) are described from marine fishes of the genus Lutjanus Bloch (Perciformes: Lutjanidae) in the northern Gulf of Mexico: P. longispicula sp. n. from the ovary of the northern red snapper L. campechanus (Poey) (type host) and silk snapper L. vivanus (Cuvier); P. latispicula sp. n. from the ovary and rarely testes of the grey snapper L. griseus (Linnaeus); and P. synagridis sp. n. (only males available) from the ovary of the lane snapper Lutjanus synagris (Linnaeus). These species are mainly characterised by the lengths of spicules (378-690 microm, 135-144 microm and 186-219 microm, respectively) and spicule shapes, structure of the distal portion of the gubernaculum and the structure of the male caudal end. These are the first valid, nominal species of gonad-infecting philometrids reported from fishes of the family Lutjanidae in the western Atlantic region.

How frequently Dirofilaria repens localizes in dog testes?

Dirofilaria repens (Spirurida: Onchocercidae) is the causative agent of canine subcutaneous dirofilariosis. Adults of D. repens localize in the subcutaneous tissues and fasciae and most of the infected animals are asymptomatic though dermatological disorders have been described sporadically. In this study, the gonads of 100 male dogs were inspected for D. repens presence from December 2020 to January 2022. All examined dogs were older than 48 months and underwent castration for several reasons. The testes and the superficial layers were examined with the aid of stereomicroscope; in addition, the blood of all included dogs was collected and analysed by Knott's test to assess the presence of microfilariae. Six out of 100 (6 %, CI95%, 2.2-12.6 %) examined dogs were positive for D. repens presence, and in all the cases the parasites were found beneath the tunica vaginalis layers. Microfilariae were found in 15 blood samples including 5 out of those 6 testes positive dogs thus resulting in a final D. repens prevalence of 16 % (CI95%: 9.4-24.7 %). No pathological changes and or lesions due to parasite presence were observed in the testes or in their layers. The present study represents the first survey focusing on the detection of D. repens in dog testes on a large sample size. Adult parasites were found in the testes of 6 out of 16 D. repens infected dogs accounting for 37.5 % of cases and suggesting as testicular localization is not uncommon. The absence of neoplastic or inflammatory lesions in D. repens infected testes suggests as these conditions do not act as attractive stimuli for the parasite localization. Finally, considering the zoonotic potential of D. repens and the large infection prevalence here reported, active surveillance in endemic areas is strongly advocated to limit the presence of reservoir and prevent further infections to animals and humans.

Differential transcriptome study on the damage of testicular tissues caused by chronic infection of T. gondii in mice.

BACKGROUND: Toxoplasma gondii is an intracellular protozoan parasite that is widely distributed in humans and warm-blooded animals. T. gondii chronic infections can cause toxoplasmic encephalopathy, adverse pregnancy, and male reproductive disorders. In male reproduction, the main function of the testis is to provide a stable place for spermatogenesis and immunological protection. The disorders affecting testis tissue encompass abnormalities in the germ cell cycle, spermatogenic retardation, or complete cessation of sperm development. However, the mechanisms of interaction between T. gondii and the reproductive system is unclear. The aims were to study the expression levels of genes related to spermatogenesis, following T. gondii infection, in mouse testicular tissue. METHODS: RNA-seq sequencing was carried out on mouse testicular tissues from mice infected or uninfected with the T. gondii type II Prugniaud (PRU) strain and validated in combination with real-time quantitative PCR and immunofluorescence assays. RESULTS: The results showed that there were 250 significant differentially expressed genes (DEGs) (P < 0.05, |log2fold change| â‰§ 1). Bioinformatics analysis showed that 101 DEGs were annotated to the 1696 gene ontology (GO) term. While there was a higher number of DEGs in the biological process classification as a whole, the GO enrichment revealed a significant presence of DEGs in the cellular component classification. The Arhgap18 and Syne1 genes undergo regulatory changes following T. gondii infection, and both were involved in shaping the cytoskeleton of the blood-testis barrier (BTB). The number of DEGs enriched in the MAPK signaling pathway, the ERK1/2 signaling pathway, and the JNK signaling pathway were significant. The PTGDS gene is located in the Arachidonic acid metabolism pathway, which plays an important role in the formation and maintenance of BTB in the testis. The expression of PTGDS is downregulated subsequent to T. gondii infection, potentially exerting deleterious effects on the integrity of the BTB and the spermatogenic microenvironment within the testes. CONCLUSIONS: Overall, our research provides in-depth insights into how chronic T. gondii infection might affect testicular tissue and potentially impact male fertility. These findings offer a new perspective on the impact of T. gondii infection on the male reproductive system.

Toxoplasma gondii infection enhances testicular steroidogenesis in rats.

The protozoan parasite Toxoplasma gondii enhances the sexual attractiveness of infected male rats and attenuates the innate fear of cat odour in infected individuals. These behavioural changes plausibly lead to greater transmission of parasites through sexual and trophic routes, respectively. Testosterone, a testicular steroid, is known to reduce fear and enhance sexual attractiveness in males. Here, we show that Toxoplasma gondii infection enhances expression of genes involved in facilitating synthesis of testosterone, resulting in greater testicular testosterone production in male rats. In several species, testosterone mediates trade-offs between sexually selected traits and life history decisions. Augmentation of testosterone synthesis by Toxoplasma gondii suggests that parasites may manipulate these trade-offs in rats.

A nuptially transmitted ichthyosporean symbiont of Tenebrio molitor (Coleoptera: Tenebrionidae).

The yellow mealworm, Tenebrio molitor, harbors a symbiont that has spores with a thick, laminated wall and infects the fat body and ventral nerve chord of adult and larval beetles. In adult males, there is heavy infection of the epithelial cells of the testes and between testes lobes with occasional penetration of the lobes. Spores are enveloped in the spermatophores when they are formed at the time of mating and transferred to the female's bursa copulatrix. Infection has not been found in the ovaries. The sequence of the nuclear small subunit rDNA indicates that the symbiont is a member of the Ichthyosporea, a class of protists near the animal-fungi divergence.

[Morphogenesis of the structural changes of the internal organs in superinvasive opisthorchiasis].

The study of the model of superinvasive opisthorchiasis (SO) in Syrian hamsters and autopsy material obtained from patients with SO, it was shown that during the early period of pulmonary disease, the lungs underwent changes developing on the basis of delayed-type hypersensitivity reactions. In the chronic phase of SO, exudative and predominant productive reactions were revealed leading to the interstitial lung disease, fibrosing alveolitis. In the heart in SO, eosinophilic cell myocarditis was found with subsequent diffuse cardiosclerosis. In the testes, sclerotic processes with atrophy of glandulocytes (Leydig cells) and sustentocytes (Sertoli cells) were detected. The structural changes in the lungs, heart and testes were induced by parasite metabolites, that are deposited in tissues in the form of granules and conglomerates.

[Complicated forms of superinvasion opisthorchiasis: pathogenetic aspects].

Superinvasion opisthorchiasis is characterized by host growth retardation and weight reduction, which are due to the cleptoeffect of Opisthorchis felineus; metabolic disturbances, proliferative processes, accelerated collagen biosynthesis and biodegradation in the organs reflect parasite-host interactions as components of the system. The productive reactions of visceral pathology are initiated by the parasite's metabolites located in the liver, lung, heart, and testes.

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.

Chloromyxum aegypticus n. sp. (Myxozoa: Chloromyxidae) infecting the testicular tissue of the Egyptian toad, Bufo regularis (Amphibia: Bufonidae), and its pathogenicity.

A new myxosporean, Chloromyxum aegypticus n. sp., is described from the testes of the Egyptian toad Bufo regularis, captured from El Mansoura locality. C. aegypticus is identified on the basis of cytology, electron microscopy and histopathology. It is distinguished from all previously reported Chloromyxum spp. by its shape, dimensions of the mature spore (9.1 ± 0.1 (9.0-9.2) µm in length × 7.9 ± 0.1 (7.8-8.0) µm in width), polar capsules, external ridge, sporoplasm nuclei, undulating suture, locality and host. Accumulation of several hundreds of plasmodia (0.8 ± 0.3 (0.5-1.1) mm in length × 0.5 ± 0.3 (0.2-0.7) mm in width) in the testes causes their enlargement. Parasites cause destruction of the seminiferous tubule cells and complete loss of spermatozoa. Since spermatogenesis stops, this seems to be a form of "parasitic castration". This is the first known record of the genus Chloromyxum in amphibian testes.

[The case of the nematode Setaria equina found in the vaginal sac of the stallion's scrotum]. / Przypadek obecnosci nicienia Setaria equina w jamie pochwowej worka mosznowego ogiera.

The nematode Setaria equina usually reside in body cavities and do not cause clinical symptoms. From time to time, however, these parasites can be located in the scrotum and spermatic cord inflicting pain and edema in these body parts. The aim of the study was to describe the case of the nematode Setaria equina found in the vaginal sac of the stallion's scrotum. During the study, thorough examination of 50 isolated testicles of 25 stallions was conducted. The horses were obtained post-slaughter from the local slaughterhouse near Krakow. In one of examined stallions, two females of Setaria equina of 80 and 110 mm in length were found attached to the surface of the testicle. A part of one of the nematode was found in the space between the surface of the testicle and the epididymis. During the examination of the testicle sections, chronic inflammation (orchitis chronica) and focal necrosis perivascularis were found.

Vascular wall injury and inflammation are key pathogenic mechanisms responsible for early testicular degeneration during acute besnoitiosis in bulls.

BACKGROUND: Bovine besnoitiosis, caused by the apicomplexan parasite Besnoitia besnoiti, is a chronic and debilitating cattle disease that notably impairs fertility. Acutely infected bulls may develop respiratory signs and orchitis, and sterility has been reported in chronic infections. However, the pathogenesis of acute disease and its impact on reproductive function remain unknown. METHODS: Herein, we studied the microscopic lesions as well as parasite presence and load in the testis (pampiniform plexus, testicular parenchyma and scrotal skin) of seven bulls with an acute B. besnoiti infection. Acute infection was confirmed by serological techniques (IgM seropositive results and IgG seronegative results) and subsequent parasite detection by PCR and histological techniques. RESULTS: The most parasitized tissue was the scrotal skin. Moreover, the presence of tachyzoites, as shown by immunohistochemistry, was associated with vasculitis, and three bulls had already developed juvenile tissue cysts. In all animals, severe endothelial injury was evidenced by marked congestion, thrombosis, necrotizing vasculitis and angiogenesis, among others, in the pampiniform plexus, testicular parenchyma and scrotal skin. Vascular lesions coexisted with lesions characteristic of a chronic infection in the majority of bulls: hyperkeratosis, acanthosis and a marked diffuse fibroplasia in the dermis of the scrotum. An intense inflammatory infiltrate was also observed in the testicular parenchyma accompanied by different degrees of germline atrophy in the seminiferous tubules with the disappearance of various strata of germ cells in four bulls. CONCLUSIONS: This study confirmed that severe acute besnoitiosis leads to early sterility that might be permanent, which is supported by the severe lesions observed. Consequently, we hypothesized that testicular degeneration might be a consequence of (i) thermoregulation failure induced by vascular lesions in pampiniform plexus and scrotal skin lesions; (ii) severe vascular wall injury induced by the inflammatory response in the testis; and (iii) blood-testis barrier damage and alteration of spermatogenesis by immunoresponse.

Trypanosoma cruzi and myoid cells from seminiferous tubules: interaction and relation with fibrous components of extracellular matrix in experimental Chagas' disease.

The main transmission route of Trypanosoma cruzi is by triatomine bugs. However, T. cruzi is also transmitted through blood transfusion, organ transplantation, ingestion of contaminated food or fluids, or is congenital. Sexual transmission, although suggested since the discovery of Chagas' disease, has remained unproven. Sexual transmission would require T. cruzi to be located at the testes and ovaries. Here we investigated whether T. cruzi is present in the gonads of mice infected with 10(4) T. cruzi trypomastigotes from the CL strain. Fourteen days after experimental infection, histopathological examination showed alterations in the extracellular matrix of the lamina propria of the seminiferous tubules. Furthermore, amastigotes were present in seminiferous tubules, within myoid cells, and in the adjacencies of the basal compartment. These results indicate that T. cruzi is able to reach seminiferous tubule lumen, thus suggesting that Chagas' disease could potentially be transmitted through sexual intercourse. Complementary studies are required to demonstrate that Chagas' disease can be transmitted by coitus.