Pathological findings in genital organs of bulls naturally infected with Besnoitia besnoiti.

Bulls chronically affected by bovine besnoitiosis can suffer from sterility. There is limited information about the distribution of Besnoitia cysts and their associated lesions within the male genital organs. This work describes the gross and histological abnormalities in the genital organs of 6 bulls chronically infected with Besnoitia besnoiti, including both clinically (n = 4) and subclinically (n = 2) affected cases. Parasitic cysts were observed in the genital organs of all the clinically affected bulls. The tissue cysts were most commonly found within the pampiniform plexus (4/4), where they were often seen within venous vascular walls and associated with vasculitis, followed by epididymis (3/4), tunica albuginea (2/4), and penis (1/4). In decreasing order of their frequency, observed abnormalities included seminiferous tubule degeneration, testicular fibrosis, testicular necrosis, lack of/or diminished numbers of spermatozoa, testicular atrophy, and Leydig cell hyperplasia. Only one of the subclinically infected bulls had few Besnoitia cysts within the pampinoform plexus, which was associated to small areas of necrosis and mineralization in the ipsilateral testicle. Results indicate that Besnoitia cysts and genital abnormalities are frequent in bulls chronically affected by bovine besnoitiosis, while they are mild and scarce in subclinically affected ones. Moreover, present data show that Besnotia-associated testicular lesions can occur without the presence of cysts within the testicular parenchyma. B. besnoiti cysts seem to have a tropism for the vascular structures of the spermatic chord, which may cause testicular abnormalities via vascular damage, reduced blood flow, and/or impaired thermoregulation and subsequently lead to the observed testicular lesions.

Case Report: Highlighting Male Genital Schistosomiasis (MGS) in Fishermen from the Southwestern Shoreline of Lake Malawi, Mangochi District.

Urogenital schistosomiasis causes morbidity within the genitalia but is underreported and infrequently examined in men. To draw attention to male genital schistosomiasis (MGS), a longitudinal cohort study was conducted among fishermen along the southwestern shoreline of Lake Malawi. A case series of five participants is presented inclusive of questionnaire interviews, parasitological examinations, ultrasonography, and provision of a standard dose (40 mg/kg) of praziquantel (PZQ) treatment at baseline, 1-, 3-, 6-, and 12-month follow-up time points. Eggs of Schistosoma haematobium were observed in urine or semen across all time points; parasitological diagnostics were bolstered by real-time PCR for Schistosoma DNA in semen and by portable ultrasonography to document putative MGS-associated morbidity. We highlight the importance of developing standard diagnostic tests for MGS and increasing the accessibility of PZQ treatment to men, especially those in at-risk endemic areas.

How can schistosome circulating antigen assays be best applied for diagnosing male genital schistosomiasis (MGS): an appraisal using exemplar MGS cases from a longitudinal cohort study among fishermen on the south shoreline of Lake Malawi.

We provide an update on diagnostic methods for the detection of urogenital schistosomiasis (UGS) in men and highlight that satisfactory urine-antigen diagnostics for UGS lag much behind that for intestinal schistosomiasis, where application of a urine-based point-of-care strip assay, the circulating cathodic antigen (CCA) test, is now advocated. Making specific reference to male genital schistosomiasis (MGS), we place greater emphasis on parasitological detection methods and clinical assessment of internal genitalia with ultrasonography. Unlike the advances made in defining a clinical standard protocol for female genital schistosomiasis, MGS remains inadequately defined. Whilst urine filtration with microscopic examination for ova of Schistosoma haematobium is a convenient but error-prone proxy of MGS, we describe a novel low-cost sampling and direct visualization method for the enumeration of ova in semen. Using exemplar clinical cases of MGS from our longitudinal cohort study among fishermen along the shoreline of Lake Malawi, the portfolio of diagnostic needs is appraised including: the use of symptomatology questionnaires, urine analysis (egg count and CCA measurement), semen analysis (egg count, circulating anodic antigen measurement and real-time polymerase chain reaction analysis) alongside clinical assessment with portable ultrasonography.

Identification and evaluation of the microbiome in the female and male reproductive tracts.

BACKGROUND: The existence of an extensive microbiome in and on the human body has increasingly dominated the scientific literature during the last decade. A shift from culture-dependent to culture-independent identification of microbes has occurred since the emergence of next-generation sequencing (NGS) techniques, whole genome shotgun and metagenomic sequencing. These sequencing analyses have revealed the presence of a rich diversity of microbes in most exposed surfaces of the human body, such as throughout the reproductive tract. The results of microbiota analyses are influenced by the technical specifications of the applied methods of analyses. Therefore, it is difficult to correctly compare and interpret the results of different studies of the same anatomical niche. OBJECTIVES AND RATIONALE: The aim of this narrative review is to provide an overview of the currently used techniques and the reported microbiota compositions in the different anatomical parts of the female and male reproductive tracts since the introduction of NGS in 2005. This is crucial to understand and determine the interactions and roles of the different microbes necessary for successful reproduction. SEARCH METHODS: A search in Embase, Medline Ovid, Web of science, Cochrane and Google scholar was conducted. The search was limited to English language and studies published between January 2005 and April 2018. Included articles needed to be original microbiome research related to the reproductive tracts. OUTCOMES: The review provides an extensive up-to-date overview of current microbiome research in the field of human reproductive medicine. The possibility of drawing general conclusions is limited due to diversity in the execution of analytical steps in microbiome research, such as local protocols, sampling methods, primers used, sequencing techniques and bioinformatic pipelines, making it difficult to compare and interpret results of the available studies. Although some microbiota are associated with reproductive success and a good pregnancy outcome, it is still unknown whether a causal link exists. More research is needed to further explore the possible clinical implications and therapeutic interventions. WIDER IMPLICATIONS: For the field of reproductive medicine, determination of what is a favourable reproductive tract microbiome will provide insight into the mechanisms of both unsuccessful and successful human reproduction. To increase pregnancy chances with live birth and to reduce reproduction-related health costs, future research could focus on postponing treatment or conception in case of the presence of unfavourable microbiota and on the development of therapeutic interventions, such as microbial therapeutics and lifestyle adaptations.

Subtle effect of Xenos vesparum (Xenidae, Strepsiptera) on the reproductive apparatus of its male host: Parasite or parasitoid?

Parasitic castration is an adaptive strategy where parasites usurp the hosts' reproductive physiology to complete their life cycle. The alterations in the host traits vary in their magnitude, from subtle changes in the host morpho-physiology and behaviour to the production of complex aberrant phenotypes, which often depend on the host gender. The strepsipteran macroparasite Xenos vesparum induces dramatic behavioural and physiological changes in its female host, the paper wasp Polistes dominula, while its effect on the male phenotype is largely unknown. In this study we investigated how a single X. vesparum parasite influences the functional morphology of P. dominula male reproductive apparatus. We performed morphometry and ultrastructure characterization of corpora allata, testes, seminal vesicles and accessory glands in parasitized and unparasitized males, and also in young and old males to control for the effect of age on the natural deterioration of these organs. Our results show that age significantly affects the development of male reproductive apparatus. A low parasite load - one parasite per host is the common prevalence in the field - has only a marginal impact on the reproductive morphology of P. dominula males, affecting quantitatively but not qualitatively the protein content of male accessory glands. Thus, in male P. dominula wasps, X. vesparum appears to behave as a true "parasite", in clear opposition to the role of "parasitoid" that it takes in female hosts where castration causes the reproductive death.

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.

Occurrence of Leishmania infantum and associated histological alterations in the genital tract and mammary glands of naturally infected dogs.

The objectives of this study were to evaluate the occurrence of Leishmania infantum in the male and female genital tract and female mammary glands of dogs and the parasite burden and to identify histological alterations associated with this protozoan. Twenty male and 20 female Leishmania-seropositive dogs with isolation of L. infantum were examined. Tissue samples of the prepuce, glans, epididymis, testes, prostate, vulva, vagina, uterus, uterine tubes, and mammary glands were analyzed by immunohistochemistry and histopathology. For parasitological culture and in situ hybridization, samples were collected from the testis, epididymis, and uterus. Additionally, seminal fluid was aspirated from the epididymis for parasitological culture. In the genital tract, 34 (85 %) dogs, including 18 males and 16 females, were positive for Leishmania. Of these, 27 (79 %) animals were symptomatic. Leishmania was detected in the mammary glands of 13 (65 %) females. L. infantum was isolated for the first time from the seminal fluid and uterus of naturally infected dogs. The parasite burden and intensity of the inflammatory reaction were greater in the prepuce and glans of males and in the vulva and mammary glands of females. In addition to inflammation, testicular degeneration, atrophy, absence of spermatogenesis, and necrosis were observed. Detection of amastigote forms in the mammary gland lumen indicates possible elimination of this parasite in milk. The frequent parasitism observed in the genital tract of infected males and females and the viability of L. infantum in seminal fluid and uterus suggest the possibility of bidirectional venereal and vertical transmission.

Revealing the kinetics of Leishmania chagasi infection in the male genital system of hamsters.

BACKGROUND: Leishmaniasis causes alterations and lesions in the genital system, which leads to azoospermia and testicular atrophy in animals during the chronic phase of the infection. The aim of this study was to reveal the kinetics of Leishmania chagasi infection in the genital system of male golden hamsters (Mesocricetus auratus). METHODS: Animals were intraperitoneally inoculated with amastigotes from L. chagasi. At different time points animals were euthanized and genital organs processed for histo-pathological, qPCR, cytokines and testosterone detection assays. RESULTS: Our results showed a high parasite load in testis, followed by an increase of pro-inflammatory cytokines IL1-ß, TNF-α and IFN-γ, and testosterone. Subsequently, IL-4 expression was upregulated and basal parasite persistence in testis was observed using the experimental approach. CONCLUSION: Extracellular amastigotes migrated to the epididymis posing as a potential major factor of parasite persistence and venereal transmission of L. chagasi infection in hamsters.

High Prevalence of Porocephalus crotali Infection on a Barrier Island (Cumberland Island) off the Coast of Georgia, with Identification of Novel Intermediate Hosts.

Porocephalus crotali is a pentastomid parasite that uses crotaline snakes as definitive hosts and a variety of rodents as intermediate hosts. A study of definitive and intermediate pentastome hosts on Cumberland Island, Georgia, revealed high prevalence of P. crotali infection in crotalid snakes as well as several mammalian species. Despite the presence of numerous nymphs in some animals, clinical signs of disease were not observed. In intermediate hosts, the liver, mesentery, and reproductive organs were most commonly infected. No gross evidence of tissue damage was noted in association with the numerous encysted nymphal pentastomes, and histopathology demonstrated minimal reaction to the encysted nymphs. Partial 18S rRNA gene sequences confirmed the parasites were P. crotali. In contrast to many previous reports in rodents, the prevalence on this barrier island was high, and this is the first report of Virginia opossums (Didelphis virginiana) and any insectivore species as intermediate hosts. Although generally not considered pathogenic, the long-term consequences of high nymph intensities on individuals deserve attention.

Urologic complications of genitourinary schistosomiasis.

OBJECTIVES: To provide systematic review of the literature on the long-standing complications of genitourinary schistosomiasis. MATERIALS AND METHODS: The PubMed literature database was searched from inception to December 2010. The following keywords were used: schistosomiasis, bilharziasis, and genitourinary. Only English language publications were utilized. RESULTS: Variable tissue reactions to bilharzial eggs with subsequent healing or progression and complications in the urinary tract mainly affect the urinary bladder and pelvic segments of the ureters. These lesions may assume an atrophic, proliferative, or neoplastic pattern. Although the pathology is usually extensive in the submucosal, all layers from the mucous membrane through deep to the perivesical or periureteral tissues may be involved. Main fixed bilharzial urologic sequelae include chronic bladder ulcers, leucoplakia, vesical granuloma, contracted bladder, bladder neck contracture, stricture ureters, and bladder carcinoma. These sequelae may lead to marked morphologic and functional changes of the urinary tract, and ultimately, mortality can follow from renal failure or bladder cancer. CONCLUSIONS: Urinary schistosomiasis is a preventable disease through nationwide snail control and mass therapy with oral antibilharzial drugs. If not properly treated, long-standing urinary complications may result in serious sequelae that may lead to mortality from renal failure or bladder cancer.

Histopathology of the reproductive system of male sheep experimentally infected with Toxoplasma gondii.

The aim of this study was to investigate the histopathological changes in reproductive system (testicles, epididymis, seminal vesicles, and prostate) of small male ruminants after Toxoplasma gondii infection. Eight sheep were inoculated with T. gondii: group I, four sheep (2.0 × 10(5) P-strain oocysts); group II, four sheep (1.0 × 10(6) RH-strain tachyzoites); and group III, two uninfected sheep maintained as control. Infection with T. gondii was confirmed by seroconversion (indirect fluorescent antibody test-IgG) in all the infected animals beginning on post-inoculation day (PID) 7. On PID 70, all the animals were euthanized and tissue samples (testicles, epididymis, seminal vesicles, and prostate) were collected and processed for histological analysis. The main changes detected were a focal mononuclear interstitial inflammatory infiltrate in the prostate and seminal vesicles; diffuse testicular degeneration associated with calcification foci and a multifocal mononuclear interstitial inflammatory infiltrate; and a mononuclear interstitial infiltrate and focal necrotic areas of the muscle fibers surrounding the seminal vesicles. The histopathological findings of this work, along with the detection of T. gondii in the examined parenchyma tissues (immunohistochemistry) and the results obtained by other authors examining different tissues, suggest that histological changes diagnosed in the reproductive system of rams infected with T. gondii are strongly suggestive of toxoplasmatic infection.

Identification of Tritrichomonas foetus pseudocysts in fresh preputial secretion samples from bulls.

Tritrichomonas foetus is a serious veterinary pathogen that causes bovine trichomonosis, a sexually transmitted disease that eventually leads to abortion and infertility. T. foetus has a simple life cycle that consists of only a trophozoitic form. During unfavorable environmental conditions, the trophozoites, which are polar and flagellated, can adopt a spherical shape and internalize their flagella. These rounded organisms are known as pseudocysts. Although it is currently assumed that T. foetus pseudocyst formation is reversible and that it represents a response to stressful conditions, there are no reports showing the presence of this form in vivo. For this reason, the aim of this study was to verify whether T. foetus pseudocysts are encountered in naturally infected bulls. Towards this goal, fresh preputial samples obtained from seven mature bulls that were naturally infected with T. foetus were analyzed using complementary techniques, such as video microscopy, fluorescence microscopy, scanning and transmission electron microscopy. The analyses revealed that approximately 55% of the parasites were in pseudocyst form in each preputial sample, whereas approximately 25% of T. foetus displayed pear-shaped bodies. Previous research demonstrated that in vitro T. foetus pseudocysts are able to divide by a budding process. Here, this division mode was observed in approximately 20% of fresh T. foetus obtained from preputial bovine samples. Thus, this study shows that in infected bulls, pseudocysts are present and occur more frequently than the pear-shaped parasites.

Detection of Toxoplasma gondii in the reproductive system of male goats.

Male goats of mating age serologically negative for Toxoplasma gondii were divided into three groups: GI--controls (placebo) (n = 2); GII--infected with 1 x 106 tachyzoites (RH strains) (n = 2); and GIII--infected with 2 x 105 oocysts (P strains) (n = 2). Clinical, hematology, parasite and serology tests and studies of parasites in the semen through bioassay and polymerase chain reaction (PCR), and in reproductive organs (bioassay) were performed to assess toxoplasma infection. Serological titers peaked at 4096 in two animal groups infected with the protozoan. The bioassays allowed an early detection of protozoa in semen samples of tachyzoite-inoculated animals. T. gondii DNA was identified through PCR in the semen in five (Days 5, 7, 28, 49, and 70) and two (both at day 56) different days post-inoculation in GII and GIII animals, respectively. It was also possible to detect T. gondii DNA in reproductive organs (prostate pool, testicles, seminal vesicle and epididymis) of goats inoculated with either tachyzoites or oocysts. The present study suggests the possibility of venereal transmission of T. gondii among goats and it should be further assessed.

Detection of Toxoplasma gondii in the reproductive system of male goats / Detecção de Toxoplasma gondii no sistema reprodutor de caprinos machos

Male goats of mating age serologically negative for Toxoplasma gondii were divided into three groups: GI - controls (placebo) (n = 2); GII - infected with 1 × 10(6) tachyzoites (RH strains) (n = 2); and GIII - infected with 2 × 10(5) oocysts (P strains) (n = 2). Clinical, hematology, parasite and serology tests and studies of parasites in the semen through bioassay and polymerase chain reaction (PCR), and in reproductive organs (bioassay) were performed to assess toxoplasma infection. Serological titers peaked at 4096 in two animal groups infected with the protozoan. The bioassays allowed an early detection of protozoa in semen samples of tachyzoite-inoculated animals. T. gondii DNA was identified through PCR in the semen in five (Days 5, 7, 28, 49, and 70) and two (both at day 56) different days post-inoculation in GII and GIII animals, respectively. It was also possible to detect T. gondii DNA in reproductive organs (prostate pool, testicles, seminal vesicle and epididymis) of goats inoculated with either tachyzoites or oocysts. The present study suggests the possibility of venereal transmission of T. gondii among goats and it should be further assessed.

Caprinos machos, em idade reprodutiva, sorologicamente negativos para Toxoplasma gondii foram distribuídos em três grupos de animais: GI (n = 2) controle (placebo), GII (n = 2) - infectado com 1 × 10(6) taquizoítos (cepa RH) e GIII (n = 2) infectado com 2 × 10(5) oocistos (cepa P). Exames clínicos, hematológicos, parasitêmicos, sorológicos, pesquisa no sêmen e em tecidos do sistema reprodutor, por meio da bioprova, e da Reação em Cadeia pela Polimerase (PCR), foram conduzidas para avaliar a infecção toxoplásmica. Os títulos sorológicos alcançaram valores máximos de 4096 nos dois grupos de animais infectados. Pela técnica da bioprova, foi possível revelar precocemente a presença do coccídio nas amostras seminais dos animais inoculados com taquizoítos. Pela PCR, foi possível identificar, no sêmen, material genético de T. gondii, em cinco (5º, 7º, 28º, 49º e 70º) e em duas (ambos ao 56º) datas experimentais pós-inoculação dos animais pertencentes aos grupos GII e GIII, respectivamente.Por esta mesma técnica, foi possível ainda isolar material genético deste protozoário, também em amostras teciduais (pool de próstata, testículo, vesícula seminal e epidídimo) dos caprinos inoculados com taquizoítos e oocistos. A presente pesquisa sugere a possibilidade da ocorrência da transmissão sexual do T. gondii na espécie caprina.

Altered expression of cytokines and sex steroid receptors in the reproductive tract of cysticercotic male mice.

Infection with Taenia crassiceps cysticerci in male mice produces an increase in serum oestradiol levels, whereas serum testosterone is abolished. Concomitantly, complete atrophy of the reproductive tract of infected male mice is observed. The present study was undertaken to determine the expression pattern of cytokines involved in steroidogenesis and sex steroid receptors in the reproductive tissues of normal and infected male mice, and relating this expression pattern to whole parasite counts, serum sex steroid levels and pathology of the reproductive tract in infected male mice. The expression of IL-4, IFN-gamma and TNF-alpha in testes and seminal vesicles was markedly increased in infected mice; however, IL-10 and IL-1beta expression was importantly decreased in the same organs. IL-2 expression in reproductive tissues was not affected by infection. The infection markedly induced the expression of androgen receptor, in both reproductive organs tested, while subtypes of oestrogen receptors were decreased in both tissues.

Natural selection and genital variation: a role for the environment, parasites and sperm ageing?

Male genitalia are more variable between species (and populations) than other organs, and are more morphologically complex in polygamous compared to monogamous species. Therefore, sexual selection has been put forward as the major explanation of genital variation and complexity, in particular cryptic female choice for male copulatory courtship. As cryptic female choice is based on differences between males it is somewhat paradoxical that there is such low within-species variation in male genitalia that they are a prime morphological identification character for animal species. Processes other than sexual selection may also lead to genitalia variation but they have recently become neglected. Here I focus on pleiotropy and natural selection and provide examples how they link genitalia morphology with genital environments. Pleiotropy appears to be important because most studies that specifically tested for pleiotropic effects on genital morphology found them. Natural selection likely favours certain genital morphology over others in various environments, as well as by reducing re-infection with sexually transmitted diseases or reducing the likelihood of fertilisation with aged sperm. Both pleiotropy and natural selection differ locally and between species so may contribute to local variation in genitalia and sometimes variation between monogamous and polygamous species. Furthermore, the multitude of genital environments will lead to a multitude of genital functions via natural selection and pleiotropy, and may also contribute to explaining the complexity of genitalia.

Bayesian estimation of Tritrichomonas foetus diagnostic test sensitivity and specificity in range beef bulls.

Accuracy of culture for diagnosis of Tritrichomonas foetus was investigated in 2832 naturally exposed range beef bulls from 124 herds. Preputial fluid samples were inoculated into the culture medium, incubated at 37 degrees C, and daily examined. Diagnostic test was evaluated using Bayesian techniques to estimate sensitivity and specificity without a gold standard. Median posterior test sensitivity was 72.04% (95% probability interval: 58.07-86.38%) and specificity was 95.37% (95% probability interval: 94.07-96.65%). Low diagnostic test accuracy may have resulted from host and/or diagnostic test procedure related factors. Under natural range conditions, more accurate methods for T. foetus diagnostic and repeated preputial samplings of bulls may be necessary on trichomonosis control programs.

[Electron microscopic study on Trichomonas vaginalis adhering to and phagocytizing male genitourinary epithelial cells].

OBJECTIVE: To observe Trichomonas vaginalis (T. vaginalis) adhering to and phagocytizing male genitourinary epithelial cells in order to study the pathogenetic mechanism of male trichomoniasis. METHODS: Cultured T. vaginalis bodies were incubated with male genitourinary epithelial cells, and then the ultrastructure was observed with transmission electron microscopy. RESULTS: T. vaginalis adhered to epithelial cells like amoeba, and formed pseudopodium or surface invagination surrounding or nibbling other parts of the epithelial cytoplasm. CONCLUSIONS: T. vaginalis has the speciality of adhering to and phagocytizing to male genitourinary epithelial cells. Genitourinary epithelial cells may be injured directly by the phagocytosis of T. vaginalis. Attention has to be paid to the correlation.

Diagnosis of trichomoniasis in 'virgin' bulls by culture and polymerase chain reaction.

The diagnostic test for Tritrichomonas foetus in bulls is microscopic examination of cultured preputial samples. Trichomonads other than T. foetus can be present in a preputial sample. Both a staining technique and a polymerase chain reaction assay were useful in differentiating between T. foetus and another trichomonad observed in samples from virgin bulls.