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Poor reproductive performance in beef cattle caused by infectious agents results in major financial losses as a result of reduced pregnancy rates and extended calving intervals. Bulls can be subclinical chronic carriers of bacterial and protozoal agents involved in cow infertility, such as Campylobacter fetus subsp. venerealis, Ureaplasma diversum, Mycoplasma bovigenitalium, Mycoplasma bovis, and Tritrichomonas foetus. Bulls harbor these microorganisms in their preputial crypts and transmit the agents to cows during natural mating. To obtain an overview of the etiologic agents in the preputial mucus of bulls, we aimed to identify, by PCR assay, C. fetus subsp. venerealis, M. bovis, U. diversum, M. bovigenitalium, and T. foetus in Brazilian bulls from farms with high infertility rates. We collected preputial mucus from 210 bulls on 18 beef cattle farms in Brazil between 2019 and 2020. We found at least one of the infectious agents that we were studying in bulls on 16 of the 18 beef cattle farms tested. We detected at least one infectious agent from 159 of 210 (76%) bulls tested, namely C. fetus subsp. venerealis, M. bovis, U. diversum, M. bovigenitalium, and T. foetus in 87 (55%), 84 (53%), 45 (28%), 28 (18%), and 1 (0.6%) animal, respectively. We found 95 bulls (60%) positive for only 1 etiologic agent (single infection) and 64 bulls (40%) carried multiple agents. Our results demonstrate the occurrence of bacterial and protozoal infectious agents that may be related to infertility in Brazilian beef cattle herds.

Bovine Trichomoniasis (BT) is an infectious disease caused by Tritrichomonas foetus that can be transmitted either sexually or by fomites. In males, the disease is asymptomatic and permanent. T. foetus has been detected in semen samples where it is able to remain viable even when frozen. The objective of this study was to investigate the presence of T. foetus in 27 samples of commercial frozen bovine semen by culture and Polymerase Chain Reaction (PCR). Samples were thawed in water at 37°C. Part of the samples was inoculated in a test tube containing Diamond’s medium and incubated at 35°C. Growth was evaluated every 24 hours via direct examination under a microscope. The other part was placed in an Eppendorf tube and frozen for later molecular analysis. After 10 days of culture, all samples were negative for T. foetus. The Quick-DNA Miniprep Kit (Zymo Research) without proteinase K was used for DNA extraction. The primers used in PCR were TRF3 and TRF4. PCR results were negative for all samples. In conclusion, bovine semen samples were negative for T. foetus in both diagnostic methods, according to the adopted methodology. RESUMO A tricomonose genital bovina (TGB), uma doença infectocontagiosa causada pelo Tritrichomonas foetus, é transmitida por via venérea e fômites contaminados. Em machos a doença é assintomática e permanente. O agente já foi encontrado em amostras de sêmen e é capaz de permanecer viável quando congelado. Este trabalho teve por objetivo investigar a presença de T. foetus em 27 amostras de sêmen bovino comercial congelado, por meio de cultivo e reação em cadeia da polimerase (PCR). As amostras foram descongeladas em água a 37ºC; parte foi inoculada em tubo de ensaio contendo meio Diamond, incubada a 35ºC com consequente avaliação de crescimento e avaliada a cada 24 horas, via exame direto em microscópio, e a outra parte foi diluída em PBS para análise molecular. Após 10 dias de cultivo, todas as amostras foram negativas. Para a detecção molecular foi utilizado o kit Quick-DNA Miniprep (Zymo Research) sem proteinase K para extração do DNA. Os iniciadores utilizados na PCR foram TRF3 e TRF4. O resultado da PCR foi negativo para todas as amostras. Conclui-se que as amostras utilizadas foram realmente negativas para a presença do patógeno em ambos os métodos diagnósticos, o que comprovou a inocuidade do sêmen testado.

Trichomonads are an order of parasitic protists which infect a wide range of hosts. The human parasite Trichomonas vaginalis and the bovine parasite Tritrichomonas foetus which also infects cats and swine are of considerable medical and veterinary importance, respectively. Since trichomonads are microaerophiles/anaerobes they are susceptible to 5-nitroimidazoles such as metronidazole. 5-nitroimidazoles are exclusively toxic to microaerophilic/anaerobic organisms because reduction, i.e. activation, of the drug can only occur in a highly reductive environment. 5-nitroimidazoles have remained a reliable treatment option throughout the last decades but drug resistance can be a problem. Clinical resistance to 5-nitroimidazoles has been studied in more detail in T. vaginalis and has been ascribed to defective oxygen scavenging mechanisms which lead to higher intracellular oxygen concentrations and, consequently, to less drug being reduced. Two enzymes, flavin reductase (FR) and NADH oxidase have been suggested to be the major oxygen scavenging enzymes in T. vaginalis. The loss, or at least an impairment of FR which reduces oxygen to hydrogen peroxide, has been proposed as the central mechanism that enables the emergence of 5-nitroimidazole resistance. In this study we explored if T. foetus also encodes a homolog of FR and if it is, likewise, involved in resistance. T. foetus was indeed found to express a FR but it was only weakly active as compared to the T. vaginalis homolog. Further, activity of FR in T. foetus was unchanged in metronidazole-resistant cell lines, ruling out that it has a role in metronidazole resistance. Finally, we measured oxygen scavenging rates in metronidazole-sensitive and –resistant cell lines and found that NADH oxidase and FR are not the major oxygen scavenging enzymes in trichomonads and that oxygen scavenging is possibly a consequence, rather than a cause of metronidazole resistance., Graphical abstract Image 1, Highlights • T. foetus encodes a flavin reductase (FR) like T. vaginalis but it is less active. • FR is not downregulated in metronidazole-resistant T. foetus as in T. vaginalis. • Flavin reductase and NADH oxidase are not the major oxygen scavenging enzymes. • Defective oxygen scavenging is not necessarily a cause of metronidazole resistance.

Trichomonas vaginalis is a neglected urogenital parasitic protist that causes 170 million cases of trichomoniasis annually, making it the most prevalent non-viral, sexually transmitted disease. Trichomoniasis treatment relies on nitroheterocyclics, such as metronidazole. However, with increasing drug-resistance, there is an urgent need for novel anti-trichomonals. Little progress has been made to translate anti-trichomonal research into commercialised therapeutics, and the absence of a standardised compound-screening platform is the immediate stumbling block for drug-discovery. Herein, we describe a simple, cost-effective growth assay for T. vaginalis and the related Tritrichomonas foetus. Tracking changes in pH were a valid indicator of trichomonad growth (T. vaginalis and T. foetus), allowing development of a miniaturised, chromogenic growth assay based on the phenol red indicator in 96- and 384-well microtiter plate formats. The outputs of this assay can be quantitatively and qualitatively assessed, with consistent dynamic ranges based on Z′ values of 0.741 and 0.870 across medium- and high-throughput formats, respectively. We applied this high-throughput format within the largest pure-compound microbial metabolite screen (812 compounds) for T. vaginalis and identified 43 hit compounds. We compared these identified compounds to mammalian cell lines, and highlighted extensive overlaps between anti-trichomonal and anti-tumour activity. Lastly, observing nanomolar inhibition of T. vaginalis by fumagillin, and noting this compound has reported activity in other protists, we performed in silico analyses of the interaction of fumagillin with its molecular target methionine aminopeptidase 2 for T. vaginalis, Giardia lamblia and Entamoeba histolytica, highlighting potential for fumagillin as a broad-spectrum anti-protistal against microaerophilic protists. Together, this new platform will accelerate drug-discovery efforts, underpin drug-resistance screening in trichomonads, and contributing to a growing body of evidence highlighting the potential of microbial natural products as novel anti-protistals., Graphical abstract Image 1

The current study was conducted on the detection of Tritrichomonas foetus in vaginal mucus of infected cows using conventional polymerase chain reaction technique (cPCR) in Nineveh province, Iraq. A total of 87 vaginal mucus samples were collected randomly from the vagina of Heifer cows of different ages (2-4, >4-6, >6 years old) and stages of pregnancy with different clinical status (early embryonic death, pyometra, abortion and healthy animals once) by washing cow’s vagina using artificial insemination pipette, DNA extraction of T. foetus was done from vaginal mucus samples, cPCR was attempt using TFR3 and TFR4 primers, Results indicated that 11 cows (12.6 %) were positive for T. foetus. The clinical status of cows demonstrated statistically significant (PT. foetusinfection was significantly (P2-4 years old) at (8%) compared to (>4-6 years old) and (>6 years old) cows. This study concluded that T. foetusinfection was an elevated percentage of infection in cows with early embryonic death and in cows (>2-4) years old. This study is the first detected T. foetus in cattle in Nineveh province.

Human trichomoniasis, caused by the pathogenic parasitic protozoan Trichomonas vaginalis, is the most common non-viral sexually transmitted disease that contributes to reproductive morbidity in affected women and possibly to prostate cancer in men. Tritrichomonas foetus strains cause the disease trichomoniasis in farm animals (cattle, bulls, pigs) and diarrhea in domestic animals (cats and dogs). Because some T. vaginalis strains have become resistant to the widely used drug metronidazole, there is a need to develop alternative treatments, based on safe natural products that have the potential to replace and/or enhance the activity of lower doses of metronidazole. To help meet this need, this overview collates and interprets worldwide reported studies on the efficacy of structurally different classes of food, marine, and medicinal plant extracts and some of their bioactive pure compounds against T. vaginalis and T. foetus in vitro and in infected mice and women. Active food extracts include potato peels and their glycoalkaloids α-chaconine and α-solanine, caffeic and chlorogenic acids, and quercetin; the tomato glycoalkaloid α-tomatine; theaflavin-rich black tea extracts and bioactive theaflavins; plant essential oils and their compounds (+)-α-bisabolol and eugenol; the grape skin compound resveratrol; the kidney bean lectin, marine extracts from algae, seaweeds, and fungi and compounds that are derived from fungi; medicinal extracts and about 30 isolated pure compounds. Also covered are the inactivation of drug-resistant T. vaginalis and T. foetus strains by sensitized light; anti-trichomonad effects in mice and women; beneficial effects of probiotics in women; and mechanisms that govern cell death. The summarized findings will hopefully stimulate additional research, including molecular-mechanism-guided inactivations and human clinical studies, that will help ameliorate adverse effects of pathogenic protozoa.

The protozoan parasite Tritrichomonas foetus causes early embryonic death in cattle which results in severe economic loss. In the United States, there are no drugs are approved for treatment of this pathogen. In this study, we evaluated in vitro anti-protozoal effects of compounds from an open access chemical library against T. foetus trophozoites. An initial high-throughput screen identified 16 compounds of interest. Further investigation revealed 12 compounds that inhibited parasite growth and 4 compounds with lethal effects. For lethal compounds, dose-response curves were constructed and the LD50 was calculated for laboratory and field strains of T. foetus. Our experiments revealed chemical scaffolds that were parasiticidal in the micromolar range, and these scaffolds provide a starting point for drug discovery efforts. Further investigation is still needed to investigate suitability of these scaffolds and related compounds in food animals. Importantly, open access chemical libraries can be useful for identifying compounds with activity against protozoan pathogens of veterinary importance., Graphical abstract Image 1, Highlights • No legal treatments are available for bovine trichomoniasis in the United States. • The open access malaria box was screened for compounds with activity against T. foetus trophozoites. • Identification of several scaffolds meriting further investigation for suitability in food animals.

Background Diarrhea is one of the most common clinical symptoms in cats and can be caused by infectious pathogens and investigation of the prevalence, co-infection and seasonality of enteropathogens are not well-established in diarrheic cats. Results Fecal samples of 1620 diarrheic cats were collected and enteropathogens were detected using real-time PCR. We retrospectively investigated the clinical features, total/seasonal prevalence, and infection patterns of enteropathogens. The positive infection rate was 82.59%. Bacterial, viral, and protozoal infections accounted for 49.3, 37.57, and 13.13% of cases, respectively. Feline enteric coronavirus (FECV) was the most common pathogen (29.37%), followed by Clostridium (C.) perfringens, Campylobacter (C.) coli, feline parvovirus, and Tritrichomonas foetus. The seasonality of enteropathogens was observed with peaks as follows: bacterial infections peaked in October, viral infections peaked in November, and protozoal infections peaked in August. Viral and protozoal infections showed differences in prevalence according to patient age. In the infection patterns, the ratios of single infections, mixed infections, and co-infections were 35.72, 9.87, and 54.41%, respectively. FECV was predominant in single infections. The most common patterns of multiple infections were C. perfringens and C. coli in mixed infections and C. perfringens and FECV in co-infections. Conclusions Infection patterns differed according to the enteropathogen species, seasonality, and age distribution in cats. The results of this study might be helpful to understand in clinical characteristics of feline infectious diarrhea. In addition, continued monitoring of feline enteropathogens is required.

Background Fecal polymerase chain reaction (PCR) testing for Tritrichomonas foetus is considered the most sensitive means for diagnosis of infection but results could be influenced by fecal collection technique and prior use of antimicrobial drugs. Objectives To establish any association between fecal collection technique or treatment history and results of fecal PCR testing for T. foetus. Animals Fecal samples from 1717 cats submitted by veterinarians between January 2012 and December 2017. Methods This study used a retrospective analysis. T. foetus PCR test results from 1808 fecal samples submitted for diagnostic testing were examined for their association with method of fecal collection and prior antimicrobial treatments. Data were collected from sample submission form. Results Positive T. foetus PCR test results were obtained for 274 (16%) cats. Fecal samples collected via fecal loop had increased probability of positive PCR test results (odds ratio [OR] 2.04, 95% confidence interval [CI] 1.31-3.17, P = .002) compared to samples collected by colonic flush. There was no association between PCR test results and treatment history, treatment type, or prior treatment with ronidazole. After an initial positive PCR test, 4/19 (21%; 95% CI 2.7%-39.4%) cats treated with ronidazole had a second positive test result. Conclusions and clinical importance Results of this study support that fecal samples collected by loop might be better for PCR diagnosis of T. foetus infection. Lack of association of ronidazole with PCR test results and a 21% all-potential-causes failure rate of ronidazole in cats with preconfirmed infection are important limitations to use of this drug.

This study reports the prevalence of potential faecal pathogens in the microbiome detected in a cohort of cats and dogs with diarrhoea in Perth, Western Australia. Records from a commercial diagnostic laboratory using faecal PCR testing between July 2014 and August 2015 were reviewed.Of 289 feline faecal samples reviewed, Salmonella spp. (1.7%), Campylobacter spp. (47.6%), Clostridium perfringens (81.3%), Giardia spp. (11.1%), Toxoplasma gondii (1.2%), Tritrichomonas foetus (4.8%), panleukopenia virus (6.5%) and coronavirus (39.5%) were detected. In dogs, Salmonella spp. (5.4%), Campylobacter spp. (36.3%), C. perfringens (85.4%), Giardia spp. (6.2%), parvovirus (9.4%), coronavirus (4.7%) and distemper virus (1.5%) were detected.

Uma pesquisa de agentes enteropatogênicos em gatos domésticos de um abrigo foi realizado como etapa da investigação das causas de diarreias crônicas intermitentes. Amostras fecais individuais de 39 gatos, com livre acesso ao ambiente externo, foram obtidas para pesquisa de helmintos através do exame parasitológico, investigação de parvovírus e coronavírus e de Cryptosporidium spp., Giardia spp. e Tritrichomonas foetus através de PCR em tempo real. Dos gatos avaliados, 30 (76,9%) foram positivos para algum ou mais de um destes agentes entéricos. Desses, 11 (28,2%) apresentaram co-infecções parasitárias. Ovos de helmintos foram observados em 48,7% dos gatos (19/30), 16 felinos (41%) foram positivos para parvovírus ou coronavírus e 25,6% (10/30) estavam infectados por protozoários. Dos positivos para protozoários, cinco apresentaram Tritrichomonas foetus (12,82%), um organismo pouco relatado no Brasil, sendo este o primeiro relato de detecção deste protozoário através de PCR em fezes de gatos no Sul do Brasil e o segundo no país. A diarreia crônica em gatos pode ser multifatorial em animais de abrigo onde a densidade populacional é elevada e os meios de controle parasitário e viral são deficitários, além das condições de higiene precárias. Os fatores associados à proliferação de doenças infecciosas em abrigos promovem o surgimento de infecções por novos patógenos como o Tritrichomonas foetus, até então pouco relatado no Brasil. This study carried out a survey about enteropathogenic agents in domestic cats’ shelter as a stage of investigation for the intermittent chronic diarrhea. Individual fecal samples from 39 cats with free access to the external environment were submitted to parasitological examination, parvovirus, and coronavirus by PCR, and Cryptosporidium spp., Giardia spp. and Tritrichomonas foetus by real-time PCR. From the cats evaluated, 30 (76.9%) were positive for one or more enteric agents, and coinfections were observed in 11 cats samples (28.2%). Helminth eggs were observed in 48.7% of cats (19/30), 16 (41%) were positive for parvovirus or coronavirus and 25.6% (10/30) were infected by protozoa. From the positives for protozoa, five cats were positive to T. foetus (12.82%). The first finding of this protozoan through PCR was in the southern Brazil, and the second was in the whole country. Chronic diarrhea in cats may be multifactorial in shelter animals where the population density is high and the control of parasitic, and viral infections are deficient. Moreover, it is due to poor hygiene conditions in these shelters. The factors associated with the proliferation of infectious diseases in shelters are correlated with new pathogens infections such as T. foetus.

Trichomoniasis is the most common nonviral sexually transmitted disease in humans, but treatment options are limited. Here, we report a resorufin-based drug sensitivity assay for high-throughput microplate-based screening under hypoxic conditions. A 5203-compound enamine kinase library and several specialized compound series were tested for the inhibition of Trichomonas growth at 10 μM with Z′ values of >0.5. Hits were rescreened in serial dilution to establish an IC50 concentration. A series of 7-substituted 7-deazaadenosine analogues emerged as highly potent anti-T. vaginalis agents, with EC50 values in the low double digit nanomolar range. These analogues exhibited excellent selectivity indices. Follow-up medicinal chemistry efforts identified an optimal ribofuranose and C7 substituent. Several nucleosides rapidly cleared cultures of T. vaginalis at a concentrations of just 2 × EC50. Preliminary in vivo evaluation in a murine trichomoniasis model (Tritrichomonas foetus) revealed promising activity upon topical administration, validating purine nucleoside analogues as a new class of antitrichomonal agents.

Programmed cell death in T. foetus does not seem to make sense at first sight; however, different mechanisms of cellular death in this unicellular organism have been observed. This review summarizes the available data related to programmed cell death already published for the cattle parasite T. foetus and attempts to clarify some crucial points to understand this mechanism found in non-mitochondriates parasites, as well as assist in future research. Important results with different treatments showed that the T. foetus can choose among different pathways how to initiate cell death. Thus, a major challenge for cellular death research remains the identification of the molecular cell death machinery of this protist, such as caspases pathway, nuclear abnormalities, morphology cell changes, cellular death in this parasite and the prospects in the future research. Although, the possibility of the existence of different pathways to cell death in trichomonads is discussed and a model for possible executioners pathways during T. foetus cell death is proposed.

Protists members of the Trichomonadidae and Tritrichomonadidae families include agents of trichomoniasis that constitute important parasitic diseases in humans and in animals of veterinary interest. One of the characteristic features of these eukaryotic microorganisms is that they contain a fibrous structure known as the costa as an important cytoskeleton structure, that differs in several aspects from other cytoskeleton structures found in eukaryotic cells. Previous proteomic analysis of an enriched costa fraction revealed the presence of several hypothetical proteins. Here we describe the localization of one of the most prevalent protein found in a, previously made, proteomic assay to confirm its presence in the costa of Tritrichomonas foetus. A peptide sequence of the hypothetical protein ARM19800.1 was selected for the production of specific polyclonal antibodies and its specificity was confirmed by Western Blotting using an enriched costa fraction. Next, the specific localization of the selected protein was evaluated by immunofluorescence and electron microscopy immunocytochemistry. Our observations clearly showed that the ARM 19800.1 protein is indeed localized in the costa and displays an almost periodic labeling pattern. Since this is the first protein identified in the costa, it was designated as costain 1. A better understanding of a structure as peculiar as the costa is of great biological and evolutionary importance due to the fact that it contains unique proteins, it may represent a possible chemotherapy target and it may correspond to antigens of interest in immunodiagnosis and/or vaccine development.

Tritrichomonas foetus (T. foetus) is the causative agent of bovine trichomoniasis that has a major impact on production costs for beef cattle producers. Immunization strategies to effectively protect against T. foetus are a high priority. Tritrichomonas foetus expresses a surface polysaccharide, beta 1-6 poly-N-acetyl glucosamine (PNAG), that is synthesized by a broad range of microbial pathogens. Vaccination with a PNAG-specific vaccine has demonstrated protection in pigs and horses. This study attempted to protect pregnant cows from an experimental T. foetus infection by prior vaccination with a PNAG-specific vaccine. Acceptable PNAG-specific antibody titers were achieved after vaccination, but antibodies elicited were non-protective against experimental challenge with T. foetus and in in vitro antibody functional bacterial killing assays., The Bovine Practitioner, Vol. 55, No. 1 (2021)

The protozoal pathogen Tritrichomonas foetus infects the colon of domestic cats and is a major cause of chronic colitis and diarrhea. Treatment failure is common, but antibiotics may improve clinical signs in a subset of cats, leading researchers to question involvement of the colonic microbiota in disease pathogenesis. Studies performed in women with venereal Trichomonas vaginalis infections have revealed that dysbiosis of host microbiota contributes to pathogenicity with similar findings also found in mice with intestinal Tritrichomonas musculis The aim of this study was to characterize differences in the fecal microbiota of cats with and without naturally occurring T. foetus infection and in a group of kittens prior to and after experimentally induced infection. Archived fecal DNA from cats undergoing testing for T. foetus infection (n = 89) and experimentally infected kittens (n = 4; at pre-, 2 weeks, and 9 weeks post-infection) were analyzed by sequencing of 16S rRNA genes. Amongst the naturally infected population, the genera Megamonas and Helicobacter were significantly increased in prevalence and abundance in cats testing positive for T. foetus infection. In the group of four experimentally infected kittens, fecal samples post-infection had significantly lower abundance of genus Dialister and Megamonas and greater abundance of the class Betaproteobacteria and family Succinivibrionaceae. We hypothesize that T. foetus promotes dysbiosis by competition for fermentable substrates used by these bacteria and that metabolic byproducts may contribute to the pathogenesis of colonic inflammation and diarrhea. Future studies are warranted for the measurement of fecal concentrations of microbial and protozoal metabolites in cats with T. foetus infection for the identification of potential therapeutic targets.

Trichomoniasis in humans, caused by the protozoal parasite Trichomonas vaginalis, is the most common non-viral sexually transmitted disease, while Tritrichomonas foetus causes trichomonosis, an infection of the gastrointestinal tract and diarrhea in farm animals and domesticated cats. As part of an effort to determine the inhibitory effects of plant-based extracts and pure compounds, seven commercially available cherry tomato varieties were hand-peeled, freeze-dried, and pounded into powders. The anti-trichomonad inhibitory activities of these peel powders at 0.02% concentration determined using an in vitro cell assay varied widely from 0.0% to 66.7% against T. vaginalis G3 (human), from 0.9% to 66.8% for T. foetus C1 (feline), and from 0.0% to 81.3% for T. foetus D1 (bovine). The organic Solanum lycopersicum var. cerasiforme (D) peels were the most active against all three trichomonads, inhibiting 52.2% (G3), 66.8% (C1), and 81.3% (D1). Additional assays showed that none of the powders inhibited the growth of foodborne pathogenic bacteria, pathogenic fungi, or non-pathogenic lactobacilli. Tomato peel and pomace powders with high content of described biologically active compounds could serve as functional food and feed additives that might help overcome adverse effects of wide-ranging diseases and complement the treatment of parasites with the anti-trichomonad drug metronidazole.

Pentatrichomonas hominis and Tritrichomonas foetus (cat genotype) have been commonly identified as intestinal trichomonads in both dogs and cats. Although P. hominis is considered as non-pathogenic protozoa in many kinds of mammals, it has the potential for zoonotic transmission. T. foetus has been recognized as the emerging causative agent of diarrhea in cats without the risk of zoonotic transmission. As pet shops are the major source of young companion animals, the present study discusses the molecular prevalence of P. hominis and T. foetus from 544 pet shop puppies and 409 kittens. The results suggest that the prevalence of P. hominis (puppies: 7.0%; kittens: 0.5%) and T. foetus (puppies: 0%; kittens: 2.4%) in pet shop young animals are low. In addition, the infections of P. hominis and T. foetus are not always associated with the clinical signs (soft or diarrhea feces). Resumo Pentatrichomonas hominis e Tritrichomonas foetus (genótipo de gato) têm sido comumente identificados como trichomonas intestinais em cães e gatos. Apesar de P. hominis ser considerado como protozoário não patogênico em muitos tipos de mamíferos, tem potencial para transmissão zoonótica. Enquanto o T. fetus foi reconhecido como o agente causador emergente de diarreia em gatos sem o risco de transmissão zoonótica. Devido às lojas de animais serem as principais fontes de filhotes de animais domésticos, o presente estudo discute a prevalência molecular e/ou o potencial zoonótico de P. hominis e T. foetus em 544 filhotes de cachorro e 409 gatos de “pet shop”. Os resultados sugerem que a prevalência de P. hominis (cães: 7,0%; gatos: 0,5%) e T. foetus (cães: 0%; gatos: 2,4%) em animais jovens de “pet shop” é baixa. Além disso, as infecções de P. hominis e T. foetus nem sempre estão associadas aos sinais clínicos (fezes moles ou diarreia).

Extracellular vesicles (EVs) have emerged as a ubiquitous mechanism for transferring information between cells and organisms across all three kingdoms of life. Parasitic unicellular eukaryotes use EVs as vehicles for intercellular communication and host manipulation. Pathogenic protozoans are able to modulate the immune system of the host and establish infection by transferring a wide range of molecules contained in different types of EVs. In addition to effects on the host, EVs are able to transfer virulence factors, drug-resistance genes and differentiation factors between parasites. In this review we cover the current knowledge on EVs from anaerobic or microaerophilic extracellular protozoan parasites, including Trichomonas vaginalis, Tritrichomonas foetus, Giardia intestinalis and Entamoeba histolytica, with a focus on their potential role in the process of infection. The role of EVs in host: parasite communication adds a new level of complexity to our understanding of parasite biology, and may be a key to understand the complexity behind their mechanism of pathogenesis. Fil: Nievas, Yésica Romina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Biotecnológicas. Instituto de Investigaciones Biotecnológicas "Dr. Raúl Alfonsín" (sede Chascomús). Universidad Nacional de San Martín. Instituto de Investigaciones Biotecnológicas. Instituto de Investigaciones Biotecnológicas "Dr. Raúl Alfonsín" (sede Chascomús); Argentina Fil: Lizarraga, Ayelen. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Biotecnológicas. Instituto de Investigaciones Biotecnológicas "Dr. Raúl Alfonsín" (sede Chascomús). Universidad Nacional de San Martín. Instituto de Investigaciones Biotecnológicas. Instituto de Investigaciones Biotecnológicas "Dr. Raúl Alfonsín" (sede Chascomús); Argentina Fil: Salas, Nehuen. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Biotecnológicas. Instituto de Investigaciones Biotecnológicas "Dr. Raúl Alfonsín" (sede Chascomús). Universidad Nacional de San Martín. Instituto de Investigaciones Biotecnológicas. Instituto de Investigaciones Biotecnológicas "Dr. Raúl Alfonsín" (sede Chascomús); Argentina Fil: Cóceres, Verónica Mabel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Biotecnológicas. Instituto de Investigaciones Biotecnológicas "Dr. Raúl Alfonsín" (sede Chascomús). Universidad Nacional de San Martín. Instituto de Investigaciones Biotecnológicas. Instituto de Investigaciones Biotecnológicas "Dr. Raúl Alfonsín" (sede Chascomús); Argentina Fil: de Miguel, Natalia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Biotecnológicas. Instituto de Investigaciones Biotecnológicas "Dr. Raúl Alfonsín" (sede Chascomús). Universidad Nacional de San Martín. Instituto de Investigaciones Biotecnológicas. Instituto de Investigaciones Biotecnológicas "Dr. Raúl Alfonsín" (sede Chascomús); Argentina

Plant-derived anthraquinones were evaluated in cell assays for their inhibitory activities against the parasitic protozoa Trichomonas vaginalis human strain G3 that causes the sexually transmitted disease trichomoniasis in women, Tritrichomonas foetus bovine strain D1 that causes sexually transmitted diseases in farm animals (bulls, cows, and pigs), Tritrichomonas foetus-like strain C1 that causes diarrhea in domestic animals (cats and dogs), and bacteria and fungi. The anthraquinones assessed for their inhibitory activity were anthraquinone, aloe-emodin (1,8-dihydroxy-3-hydroxymethylanthraquinone), anthrarufin (1, 5-dihydroxyanthraquinone), chrysazin (1,8-dihydroxyanthraquinone), emodin (1,3,8-trihydroxy-6-methylanthraquinone), purpurin (1,2,4-trihydroxyanthraquinone), and rhein (1,8-dihydroxy-3-carboxyanthraquinone). Their activities were determined in terms of IC50 values, defined as the concentration that inhibits 50% of the cells under the test conditions and calculated from linear dose response plots for the parasitic protozoa, and zone of inhibition for bacteria and fungi, respectively. The results show that the different substituents on the anthraquinone ring seem to influence the relative potency. Analysis of the structure–activity relationships in protozoa indicates that the aloe-emodin and chrysazin with the highest biological activities merit further study for their potential to help treat the diseases in women and domestic and farm animals. Emodin also exhibited antifungal activity against Candida albicans. The suggested mechanism of action and the additional reported beneficial biological properties of anthraquinones suggest that they have the potential to ameliorate a broad spectrum of human diseases.

Bovine Trichomonosis, an endemic sexually transmitted disease in countries with extensive livestock and natural service, represents one of the most common causes of reproductive failure. 5-nitroimidazoles and their derivatives are used for its treatment, mainly metronidazole (Mz). The emergence of resistance mechanisms adopted by the parasites against the drug and failure of the treatments suggest the need to investigate susceptibility and obtain current values. The available information of in vitro susceptibility of these drugs comes from the use of a diversity of methodologies and criteria, especially observation of the mobility of the parasite under the optical microscope to evaluate its viability. These techniques are arduous and time consuming and lead to a subjective assessment dependent on the operator, the methodology used, and the morphology adopted by the protozoan. In this sense, flow cytometry has proven to be a fast and efficient method to evaluate viability in other protozoa. The aim of this study was to evaluate the in vitro susceptibility of six bovine isolates of Tritrichomonas foetus to Mz in aerobic (AC) and anaerobic (ANC) conditions by means of the calculation of the 50% inhibitory concentration (IC 50 ), by flow cytometry, and also to analyze minimum lethal concentration (MLC) by means of recovery tests post-treatment in vitro. IC 50 values ranged from 1.06 to 1.25 μM in ANC and from 1.44 to 3.03 μM in AC, these being the only ones reported at 48 h for these protozoa. With respect to MLC at 48 h, the results were from 3.67 to 7.35 μM in ANC, and from 7.35 to 14.7 μM for AC, where two isolates (Tf0 and Tf2) for AC and one (Tf2) for ANC showed higher values than those described in the literature. Flow cytometry has proven to be an effective, rapid and objective methodology and very useful in susceptibility tests. The data obtained through these tests allow us to describe the susceptibility exhibited by these protozoa, this being valuable information when establishing dosages in Mz treatments. Fil: Rivero, María Belén. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet Noa Sur. Instituto Multidisciplinario de Salud, Tecnología y Desarrollo. - Universidad Nacional de Santiago del Estero. Instituto Multidisciplinario de Salud, Tecnología y Desarrollo; Argentina. Universidad Nacional de Santiago del Estero. Facultad de Ciencias Médicas; Argentina Fil: Luque, Melchor Emilio. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet Noa Sur. Instituto Multidisciplinario de Salud, Tecnología y Desarrollo. - Universidad Nacional de Santiago del Estero. Instituto Multidisciplinario de Salud, Tecnología y Desarrollo; Argentina. Universidad Nacional de Santiago del Estero. Facultad de Ciencias Médicas; Argentina. Universidad Nacional de Santiago del Estero. Facultad de Agronomía y Agroindustrias; Argentina Fil: Abdala, María Eugenia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet Noa Sur. Instituto Multidisciplinario de Salud, Tecnología y Desarrollo. - Universidad Nacional de Santiago del Estero. Instituto Multidisciplinario de Salud, Tecnología y Desarrollo; Argentina. Universidad Nacional de Santiago del Estero. Facultad de Ciencias Médicas; Argentina. Universidad Nacional de Santiago del Estero. Facultad de Agronomía y Agroindustrias; Argentina Fil: Luna, Bruno Elias. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet Noa Sur. Instituto Multidisciplinario de Salud, Tecnología y Desarrollo. - Universidad Nacional de Santiago del Estero. Instituto Multidisciplinario de Salud, Tecnología y Desarrollo; Argentina Fil: Di Lullo, David. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet Noa Sur. Instituto Multidisciplinario de Salud, Tecnología y Desarrollo. - Universidad Nacional de Santiago del Estero. Instituto Multidisciplinario de Salud, Tecnología y Desarrollo; Argentina Fil: Carranza, Pedro Gabriel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet Noa Sur. Instituto Multidisciplinario de Salud, Tecnología y Desarrollo. - Universidad Nacional de Santiago del Estero. Instituto Multidisciplinario de Salud, Tecnología y Desarrollo; Argentina. Universidad Nacional de Santiago del Estero. Facultad de Ciencias Médicas; Argentina. Universidad Nacional de Santiago del Estero. Facultad de Agronomía y Agroindustrias; Argentina Fil: Rivero, Fernando David. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet Noa Sur. Instituto Multidisciplinario de Salud, Tecnología y Desarrollo. - Universidad Nacional de Santiago del Estero. Instituto Multidisciplinario de Salud, Tecnología y Desarrollo; Argentina. Universidad Nacional de Santiago del Estero. Facultad de Ciencias Médicas; Argentina. Universidad Nacional de Santiago del Estero. Facultad de Agronomía y Agroindustrias; Argentina

An 8-year-old, spayed female Rottweiler dog, under immunosuppressant treatment for protein-losing enteropathy (PLE) and intestinal lymphangiectasia, was presented for anorexia, poor general conditions and episodes of diarrhea. A subcutaneous mass between the caudal abdominal mammary glands was found. A fine-needle aspiration cytology was performed and revealed the presence of inflammatory cells mixed with pear- or round-shaped microorganisms with cytomorphological features of flagellated protozoan trophozoites, belonging to Trichomonadida order. The final diagnosis of Tritrichomonas foetus infection was achieved with molecular analysis (Real-Time PCR) and sequencing. T. foetus has often been reported as a pathogenic agent in cattle and cats, and rarely reported in dogs, where its pathogenetic role is poorly understood. To the author's knowledge, this is the first case of T. foetus infection described in a location different from the gastrointestinal tract in a dog.

Background Tritrichomonas foetus is the etiologic agent of the sexually transmitted disease Bovine Trichomonosis (BT).In Argentina, BT is endemic and represents a relevant health problem that causes reproductive inefficiency in cattle andlarge economic losses. Metronidazole is the drug of choice in the treatment of BT. Treatment has been associated with atemporary resolution of the clinical signs but is not able to control the disease. In recent years, the apparition of in vivo andin vitro aerobic and anaerobic resistance leading to ineffective treatments has been reported.Aims Thus, the aim of the present study was to explore the susceptibility of six different isolates of T. foetus under aerobic(AC) and anaerobic (ANC) conditions.Results and discussion Six isolates of T. foetus were obtained from samples of preputial smegma of bovine origin. Values ofminimum lethal concentration and minimum inhibitory concentration were higher than those observed in other works andrepresent current data in Argentina and provide information to establish new treatment protocols. Fil: Rivero, María Belén. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet Noa Sur. Instituto Multidisciplinario de Salud, Tecnología y Desarrollo. - Universidad Nacional de Santiago del Estero. Instituto Multidisciplinario de Salud, Tecnología y Desarrollo; Argentina. Universidad Nacional de Santiago del Estero. Facultad de Ciencias Médicas; Argentina Fil: Luque, Melchor Emilio. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet Noa Sur. Instituto Multidisciplinario de Salud, Tecnología y Desarrollo. - Universidad Nacional de Santiago del Estero. Instituto Multidisciplinario de Salud, Tecnología y Desarrollo; Argentina. Universidad Nacional de Santiago del Estero. Facultad de Ciencias Médicas; Argentina. Universidad Nacional de Santiago del Estero. Facultad de Agronomía y Agroindustrias; Argentina Fil: Abdala, María Eugenia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet Noa Sur. Instituto Multidisciplinario de Salud, Tecnología y Desarrollo. - Universidad Nacional de Santiago del Estero. Instituto Multidisciplinario de Salud, Tecnología y Desarrollo; Argentina. Universidad Nacional de Santiago del Estero. Facultad de Ciencias Médicas; Argentina. Universidad Nacional de Santiago del Estero. Facultad de Agronomía y Agroindustrias; Argentina Fil: Luna, Bruno Elias. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet Noa Sur. Instituto Multidisciplinario de Salud, Tecnología y Desarrollo. - Universidad Nacional de Santiago del Estero. Instituto Multidisciplinario de Salud, Tecnología y Desarrollo; Argentina Fil: Di Lullo, David. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet Noa Sur. Instituto Multidisciplinario de Salud, Tecnología y Desarrollo. - Universidad Nacional de Santiago del Estero. Instituto Multidisciplinario de Salud, Tecnología y Desarrollo; Argentina Fil: Echaide, Ignacio Eduardo. Instituto Nacional de Tecnología Agropecuaria. Centro Regional Santa Fe. Estación Experimental Agropecuaria Rafaela; Argentina Fil: Carranza, Pedro Gabriel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet Noa Sur. Instituto Multidisciplinario de Salud, Tecnología y Desarrollo. - Universidad Nacional de Santiago del Estero. Instituto Multidisciplinario de Salud, Tecnología y Desarrollo; Argentina. Universidad Nacional de Santiago del Estero. Facultad de Ciencias Médicas; Argentina. Universidad Nacional de Santiago del Estero. Facultad de Agronomía y Agroindustrias; Argentina Fil: Rivero, Fernando David. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet Noa Sur. Instituto Multidisciplinario de Salud, Tecnología y Desarrollo. - Universidad Nacional de Santiago del Estero. Instituto Multidisciplinario de Salud, Tecnología y Desarrollo; Argentina. Universidad Nacional de Santiago del Estero. Facultad de Ciencias Médicas; Argentina. Universidad Nacional de Santiago del Estero. Facultad de Agronomía y Agroindustrias; Argentina

Tritrichomonas foetus is an important pathogen of the bovine reproductive tract causing early embryonic death and abortion in cows and persistent, asymptomatic infection in bulls. PCR detection methods have greatly enhanced diagnostic accuracy over culture; however, pre-analytical sample handling is just as critical as technical performance in detecting this pathogen and is not well studied. The purpose of this study was to investigate the effects of biological materials present in the prepuce on PCR detection of T. foetus in a variety of collection media. Simulated preputial samples were created using InPouchTM (IP) media, lactated ringers solution (LRS), or sterile saline (SAL); inoculated with low numbers of one of three T. foetus strains; and spiked with either blood, semen, urine, or sham treatment. Samples were transported to the lab, placed in growth media (LRS and SAL samples), incubated, and tested for T. foetus by PCR. Samples containing urine had statistically significantly greater mean Ct values (P = 0.008) than samples containing other materials, seen most dramatically in IP (P < 0.0001.) Urine contamination resulted in significantly (P = 0.037) fewer samples being identified as “positive” for T. foetus. Overall, SAL collections also had significantly higher mean Ct than IP or LRS (P < 0.001), and were less likely (P = 0.018) to results in classification as a “positive” sample. Results of this study indicate that collection media and biological materials can affect T. foetus PCR detection. The presence of urine in preputial samples can result in false negative results, while blood had no detrimental effects.

Graphical abstract, Highlights • Marked necrotising to lymphoplasmacytic Tritrichomonas foetus-induced colitis. • Microabscesses in crypts of the colon with T. foetus. • Coinfection of Tritrichomonas foetus, FPV and FCoV., Tritrichomonas foetus is a flagellate protist which commonly causes a waxing and waning large bowel diarrhoea in young cats. We report severe T. foetus infection of the colon, cecum and ileum with concurrent feline enteric coronavirus (FCoV) and feline panleukopenia virus (FPV) in a 3-month-old Bengal kitten with an 8-day history of vomiting, diarrhoea, failure to thrive and coughing. Protozoa filling the lumen and crypts and occasional invading into lamina propria were identified within the affected colon and confirmed by PCR as T. foetus ‘feline genotype’. Assessment of faeces by PCR revealed concurrent infection with FCoV and FPV. It is possible that immunosuppression by FPV played a role in the unprecedented T. foetus infection intensity observed histologically. Studies during and after resolution of FPV infection, will be critical to determine if T. foetus co-infection affects long-term prognosis of FPV survivors.

Bovine genital campylobacteriosis (BGC) and bovine genital trichomonosis (BGT) are globally widespread, predominantly sexually transmitted diseases that mainly affect herds in which natural mating takes place. Bulls serve as good epidemiological indicators for studying the causative agents of these diseases in cattle herds. The objective of the present work was to determine the prevalence of BGC and BGT among bulls sent to slaughterhouses in southern Minas Gerais, Brazil. Samples of preputial smegma from 200 sexually mature, non-castrated bulls of several ancestries were collected in 2013 from four regional slaughterhouses. The polymerase chain reaction (PCR) was employed to detect Tritrichomonas foetus, Campylobacter fetus subsp. fetus, and C. fetus subsp. venerealis. Isolation also was used aiming to diagnosis of BGT. All smegma samples were negative in culture for T. foetus. Molecular tests revealed 8% (16/200) of animals to be positive for T. foetus, 17.5% (35/200) for Campylobacter. fetus subsp. fetus, and 13.5% (27/200) for C. fetus subsp. venerealis. The results of the present study indicate that T. foetus, C. fetus subsp. venerealis, and C. fetus subsp. fetus are present among bulls slaughtered in southern Minas Gerais, and that BGC and BGT occur in this region. These diseases therefore need to be considered during differential diagnosis of reproductive diseases affecting cattle herds in which natural mating is employed. A Campilobacteriose Genital Bovina (CGB) e a Tricomonose Genital Bovina (TGB) são doenças de transmissão predominantemente venérea, de ampla disseminação mundial, acometendo principalmente rebanhos submetidos à monta natural. Touros constituem bons indicadores epidemiológicos para a pesquisa destes agentes nos rebanhos. O objetivo deste estudo foi determinar a prevalência da CGB e da TGB em touros encaminhados a abatedouros da região sul de Minas Gerais, Brasil. Amostras de esmegma prepucial de 200 touros (machos não castrados em idade compatível com a monta) de diversas genealogias foram coletadas em quatro abatedouros da região, no ano de 2013. A reação em cadeia da polimerase (PCR) foi utilizada para detecção dos patógenos Tritrichomonas foetus, Campylobacter fetus subsp. fetus e Campylobacter fetus subsp. venerealis. O isolamento também foi utilizado para o diagnóstico da TGB. Todas as amostras analisadas apresentaram resultado negativo na cultura para isolamento de T. foetus. Por meio dos testes moleculares, verificaram-se prevalências de 8% (16/200) de animais positivos para T. foetus; 17,5% (35/200) para C. fetus subsp. fetus e de 13,5% (27/200) para C. fetus subsp. venerealis. Os dados do presente estudo apontaram a presença de T. foetus, C. fetus subsp. venerealis e C. fetus subsp. fetus em touros abatidos na região sul de Minas Gerais, evidenciando a ocorrência da TGB e da CGB nesta região e a necessidade de inclusão destas enfermidades no diagnóstico diferencial das doenças da reprodução em rebanhos bovinos submetidos à monta natural.

The microaerophilic parasites Entamoeba histolytica, Trichomonas vaginalis, and Giardia lamblia jointly cause hundreds of millions of infections in humans every year. Other microaerophilic parasites such as Tritrichomonas foetus and Spironucleus spp. pose a relevant health problem in veterinary medicine. Unfortunately, vaccines against these pathogens are unavailable, but their microaerophilic lifestyle opens opportunities for specifically developed chemotherapeutics. In particular, their high sensitivity towards oxygen can be exploited by targeting redox enzymes. This review focusses on the redox pathways of microaerophilic parasites and on drugs, either already in use or currently in the state of development, which target these pathways.

Highlights • Most cats in shelters, sanctuaries, foster homes, and trap-neuter-return programs carried at least one enteropathogen. • Tritrichomonas foetus and coronavirus were more common in cats with diarrhea. • Other bacterial, viral, protozoal, and helminth enteropathogens were present in cats with normal feces and diarrhea., The objective of this study was to determine the prevalence of enteropathogens in cats with and without diarrhea in four different models for managing unowned cats: short-term animal shelter, long-term sanctuary, home-based foster care, and trap-neuter-return. Fecal samples from 482 cats, approximately half of the cats with normal fecal consistency and half with diarrhea, were tested by zinc sulfate centrifugation and by real-time PCR for a panel of enteropathogens. At least one enteropathogen of feline or zoonotic importance was detected in a majority of cats, regardless of management model. For most enteropathogens, the presence or absence of diarrhea was not significantly associated with infection, the exceptions being Tritrichomonas foetus in sanctuary cats with diarrhea (26%) and normal fecal consistency (10%), respectively (P ≤ 0.04), and feline coronavirus in foster cats (80% and 58%) (P ≤ 0.001). The types of enteropathogens detected were related to the type of management model, e.g., viral and protozoal infections were most common in shelters, sanctuaries, and foster homes (confinement systems), whereas helminth infections were most common in trap-neuter-return programs (free-roaming cats). These results suggest that management practices for unowned cats are inadequate for control of enteropathogens and that the presence of diarrhea is a poor indicator of enteropathogen carriage. Risk-management strategies to reduce transmission to people and other animals should focus on sanitation, housing, compliance with preventive care guidelines, periodic surveillance, response to specific enteropathogens, humane population management of free-roaming community cats, public health education, and minimizing the duration and number of cats in mass confinement.

Tritrichomonas foetus causes chronic large-bowel diarrhoea in cats, complicated not only by its chronicity but for its difficulty to treat, with few registered or effective products available, subsequently resulting in ronidazole frequently used off-label. The extent of T. foetus resistance to ronidazole is unknown. To demonstrate in vitro resistance of feline T. foetus to ronidazole and the self-resolution of the infection, we isolated novel T. foetus “feline” genotype strains from two cats in 2017 from the same household and monitored these cats over a three-year period. Axenic cultures of T. foetus from the cats were tested for in vitro resistance to metronidazole and ronidazole using anaerobic and aerobic culture assays. The minimum lethal concentration for the novel strains after incubation in aerobic conditions for 48 h were 50–100 μg/ml and 6.25–12.50 μg/ml for metronidazole and ronidazole, respectively. For susceptible T. foetus strains, the minimum lethal concentrations ranged from 1.56–12.50 μg/ml for metronidazole, and 0.39–3.13 μg/ml for ronidazole. The self-resolution of the infection was assessed using repeat diagnostic qPCR and culture. Whilst positive in 2018, the cat samples are no longer returning positive qPCR results in 2020 and 2021, indicating self-resolution over this period. This study demonstrated resistance to ronidazole for the first time in Australian cats and infection self-resolution without antimicrobial intervention is demonstrated. This study provides clinicians with the evidence to reduce the use of off-label ronidazole, and advice on the resolution timeframe for cats in single households.

Background Tritrichomonads like porcine Tritrichomonas foetus (previously named Tritrichomonas suis), can commensally live in nasal cavity of pigs, but it is rare to cause pulmonary tritrichomonosis. Case presentation A 40-day-old piglet was presented for persistent labor breathing and diagnosed with parasite infections in the lung by analysis of bronchoalveolar lavage (BAL) under microscope. By taking advantage of next-generation sequencing approach, we found 9611 homologous tags belonging to 50 annotated genes of tritrichomonads by analysis of mRNA of the bronchoalveolar lavage with the parasite infection. Furthermore, RT-PCR and DNA sequencing analysis confirmed the presence of the tritrichomonad. Findings Here, we report a case of pulmonary tritrichomonosis in a pig. By taking advantage of next-generation sequencing approach, we found 9611 homologous tags belonging to 50 annotated genes of tritrichomonads by analysis of mRNA of the bronchoalveolar lavage with the parasite infections. Furthermore, RT-PCR and DNA sequencing analysis confirmed the presence of the tritrichomonad. Conclusion Our results demonstrate that tritrichomonads like porcine Tritrichomonas foetus can cause lung infections of pigs and reveal that next-generation sequencing is potential to identify rare diseases like pulmonary tritrichomonosis in clinical. Electronic supplementary material The online version of this article (doi: 10.1186/s12917-017-1242-x) contains supplementary material, which is available to authorized users.

The pathological and molecular findings associated with Histomonas meleagridis are described in a leucistic Indian peafowl (Pavo cristatus) from Southern Brazil. The most significant gross findings were multifocal necrotizing hepatitis and diphtheric typhlitis. Histopathologic evaluation of the liver, ceca, kidney, spleen, and small intestine revealed systemic histomoniasis (SH) associated with intralesional and intravascular accumulations of histomonad organisms consistent with H. meleagridis. PCR was used to amplify the DNA of H. meleagridis from the liver, ceca, small intestine, spleen, lungs, and kidneys. Direct sequencing and phylogenetic analyses confirmed that the isolate of the flagellated trichomonad identified from this investigation is more phylogenetically related to H. meleagridis than Tetratrichomonas gallinarum, Tritrichomonas foetus, and Dientamoeba fragilis. These results confirmed the occurrence of SH in this peafowl and add to the diagnosis of this disease in birds from Brazil. This report might represent the first complete identification of spontaneous histomoniasis in a peafowl due to pathological and molecular characteristics and one of the few documented cases of SH in non-commercial birds.

The prevalence of bovine trichomonosis (BT) in TN bulls was estimated through both active screening of bulls and review of previous laboratory records. During the active bull screening, preputial smegma specimens were collected from 458 TN beef bulls at 2 cattle slaughterhouses and 2 stockyards, which serve most beef bulls in TN, between March 2014 and June 2015. Each specimen was cultured for Tritrichomonas foetus (T. foetus) as well as evaluated microscopically every other day for seven days for any protozoa resembling T. foetus. An aliquot of the culture media from each specimen was used for DNA extraction and subsequent qPCR testing. Two specimens were considered suspect on microscopic evaluation, but all specimens were negative for T. foetus on qPCR. This suggests that the 2 specimens were most likely contaminated by fecal trichomonads. Retrospectively, 1979 T. foetus test records from 2 major TN diagnostic laboratories were reviewed between October 2013 and September 2016. True prevalence of BT in TN beef bulls was estimated at

Trichomonas vaginalis, the most common nonviral sexually transmitted parasite, causes ∼283 million trichomoniasis infections annually and is associated with pregnancy complications and increased risk of HIV-1 acquisition. The antimicrobial drug metronidazole is used for treatment, but in a fraction of clinical cases, the parasites can become resistant to this drug. We undertook sequencing of multiple clinical isolates and lab derived lines to identify genetic markers and mechanisms of metronidazole resistance. Reduced representation genome sequencing of ∼100 T. vaginalis clinical isolates identified 3,923 SNP markers and presence of a bipartite population structure. Linkage disequilibrium was found to decay rapidly, suggesting genome-wide recombination and the feasibility of genetic association studies in the parasite. We identified 72 SNPs associated with metronidazole resistance, and a comparison of SNPs within several lab-derived resistant lines revealed an overlap with the clinically resistant isolates. We identified SNPs in genes for which no function has yet been assigned, as well as in functionally-characterized genes relevant to drug resistance (e.g., pyruvate:ferredoxin oxidoreductase). Transcription profiles of resistant strains showed common changes in genes involved in drug activation (e.g., flavin reductase), accumulation (e.g., multidrug resistance pump), and detoxification (e.g., nitroreductase). Finally, we identified convergent genetic changes in lab-derived resistant lines of Tritrichomonas foetus, a distantly related species that causes venereal disease in cattle. Shared genetic changes within and between T. vaginalis and Tr. foetus parasites suggest conservation of the pathways through which adaptation has occurred. These findings extend our knowledge of drug resistance in the parasite, providing a panel of markers that can be used as a diagnostic tool.

Bovine trichomoniasis is a venereal disease caused by the protozoa Tritrichomonas foetus. The aim of the study was to investigate the prevalence of T. foetus in cattle and associated risk factors. A total of 200 samples from 183 cows and 17 bulls were collected from Veterinary Teaching Hospital, Bangladesh Agricultural University; Mymensingh district and Regional station of Bangladesh Livestock Research Institute in Shahjadpur, Sirajgonj during the period from March 2015 to May 2016. Among 200 cattle, 65 were indigenous, 100 were Holstein cross and 35 were Shahiwal cross bred. The collected samples were vaginal and uterine washing, aborted foetus and semen. The samples were collected from vagina and uterus by flashing with phosphate buffer solution by artificial insemination tube connected with 20 ml syringe. The samples were examined for T. foetus under microscope. The samples were examined on the same day of collection. Out of 200 samples examined, 3 (1.5%) were found infected with T. foetus protozoa. Among these 3 positive cases of tritrichomoniasis, one was detected in aborted foetal materials and other two were detected in vaginal washings. We did not find any T. foetus in the semen samples from the bulls. Therefore, bulls represent zero prevalence of T. foetus. During the study period the examined animals were categorized into three age groups viz. 40, 60 and 100 cattle for below 3 years, over 3 years to below 6 years and over 6 years to ten years, respectively. The prevalence rate was found in adult cattle over 6 years (>6–10 years) was 2.00%, over 3 years (>3–6 years) was 1.66% and below 3 years (

Tritrichomonas foetus is a causative agent of feline trichomonosis, resulting in large-bowel diarrhea in cats. Feline trichomonosis has been reported in the USA, Europe and some Asian countries but there is limited information for Thailand. This study investigated the prevalence of T. foetus infection in cats in the Bangkok Metropolitan Area and evaluated the in vitro efficacy of metronidazole (MDZ) and ronidazole (RDZ) against T. foetus Thai isolates. In total, 215 fecal samples were collected from 121 owned cats and 94 stray cats. All fecal samples were cultivated in InPouch™ TF-Feline medium. Afterward, polymerase chain reaction (PCR) assays targeting the 5.8S rRNA gene, the ITS regions and DNA sequencing were used for the confirmation of T. foetus. The overall prevalence of T. foetus infection was 4.18% (9/215) based on cultivation and PCR. The sequencing results showed 99–100% homology to T. foetus sequences from GenBank. The average minimal lethal concentrations (MLCs) of MDZ were 333.33 and 66.67 μg/ml at 24 and 48 h, respectively. The average MLCs of RDZ were 29.16 and 12.5 μg/ml at 24 and 48 h, respectively. The MLC of the MDZ results revealed that T. foetus Thai isolates had a tendency to be MDZ-resistant. To the best of the authors' knowledge, this study was the first using in vitro cultivation and molecular techniques to report and confirm the presence of T. foetus in cats living in the Bangkok Metropolitan Area. Further studies are needed to determine the genuine infection rate of T. foetus in a greater population sample and the infection status in cats with signs of diarrhea in Thailand.

PICO question In cats infected with Tritrichomonas foetus, does treatment with oral ronidazole compared to an alternative antiprotozoal treatment or placebo result in successful resolution of clinical signs and eradication of disease? Clinical bottom line Ronidazole use appears to be efficacious in eradicating infection with Tritrichomonas foetus and resolving diarrhoea associated with infection. A dose range of 30–50 mg/kg 12–24 hourly has been suggested, with evidence suggesting that a dose of 30 mg/kg 24 hourly for 14 days may be effective. However, some cats may require higher doses and some may not respond to treatment, and relapse may occur during a protracted period following completion of the treatment course. Neurological side effects appear to be uncommon but may occur with doses of 30 mg/kg and above. A total of six studies are reviewed: Three randomised, controlled studies, one cohort study and two case series (one retrospective). Findings indicate efficacy of ronidazole treatment in eradicating infection and resolving diarrhoea, however many studies involved small sample sizes and limited follow-up. Therefore, evidence to support the use of ronidazole in Tritrichomonas foetus infected cats remains relatively limited.

There is evidence supporting clearance of the Tritrichomonas foetus in young bulls. The reasons for mature bulls becoming chronic carriers and resolution of venereal infections in young bulls have not been illuminated. It has been hypothesized that there is variation in the inflammatory response in the reproductive tract as the bull ages. Hence, the objectives of this study were to evaluate the immune cell population in the normal bovine penile and preputial epithelium and to examine any differences in cell populations between young and mature age groups. Our hypothesis was that there are significant differences in immune cell types within the penile and preputial epithelium and dermis of young versus mature bulls., American Association of Bovine Practitioners Proceedings of the Annual Conference, 2019

This chapter considers protozoa are : Entamoeba histolytica, E. histo-lytica-like amebae (Laredo type), E. hartmanni, E. coli, E. gingivalis and E. invadens; Trichomonas vaginalis, T. tenax, T. gallinae, Tritrichomonas foetus and Pentatrichomonas hominis; Giardia lamblia, and G. duodenalis.. In vitro cultivation of protozoa, free-living or parasitic, is presently more an art than a science, and whether the reverse will be true some day is a moot point. The taxonomic scheme followed throughout this chapter is one recently adopted by the Committee on Systematics and Evolution of the Society of Protozoologists. The life cycle of E. histolytica is as follows: ingested mature cysts excyst in the lower ileum. Colonization occurs throughout the colon. Encystment takes place in the lower colon and the cysts leave the host via feces. The history of the cultivation of Entamoeba is dealt with in three sections: xenic, mon-oxenic, and axenic culture systems. Those studies considered influential in advancing the technology will receive most attention.

Bovine trichomoniasis is a venereal disease caused by the flagellate protozoan Tritrichomonas foetus. Infection is related to low conception rates and would have significant impact on calf crop. The state of La Pampa started in 2006 an unprecedented mandatory control program for eradication of bovine trichomoniasis. The compulsory participation of all cattle producers and the yearly control of every bull should be followed by culling of every positive animal. This retrospective study on data from eight years of the control plan showed that 80% of farms had a single year of positive tests. In these farms, positive tests showed a strong decay of disease during the first years that reached a baseline by 2012. A non negligeable proportion of positive bulls in this group can be attributed to false positive tests. Oppositely, farms with two or more years of positive diagnosis accounted for a great proportion of recent cases. These farms were more likely related to less intensive control measures. The non exclusion of carrier bulls is the major factor contributing to the persistance of bovine trichomoniasis. Fil: Oyhenart, Jorge. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Ciencias de la Tierra y Ambientales de La Pampa. Universidad Nacional de La Pampa. Facultad de Ciencias Exactas y Naturales. Instituto de Ciencias de la Tierra y Ambientales de La Pampa; Argentina

Practical relevance: Trichomonosis of the large intestine of the cat was described as a cause of chronic diarrhea over 20 years ago. The trichomonad was identified as Tritrichomonas foetus, with a genotype that is distinct from venereal T foetus of cattle. Clinical challenges: Despite multiple means for diagnosis of the infection, including light microscopy, protozoal culture and PCR amplification using species-specific primers, tests with even greater sensitivity are needed. Feline trichomonosis is resistant to all commonly used antiprotozoal drugs. Ronidazole is currently the only drug demonstrated to be effective in eliminating the infection from cats; however, this drug has a narrow safety margin and clinical resistance is increasingly recognized. The more we learn about trichomonosis in cats, the more complicated and controversial the infection has become, ranging from what we should call the organism to whether we should even bother trying to treat it. Global importance: Feline trichomonosis is recognized to occur worldwide and is regarded as one of the most common infectious causes of colitis in the domestic cat. The infection is widespread in catteries and shelters; and, while remission of diarrhea may occur over time, persistence of the infection is common. Evidence base: This review provides a comprehensive examination of what is currently known about feline trichomonosis and pinpoints areas, based on the authors’ opinion, where further research is needed.

The objective of this study was to determine the level and duration of IgG antibodies induced against killed whole Tritrichomonas foetus and T foetus-purified surface antigen (TF1.17) in serum, vaginal, and uterine secretions after systemic immunization of beef cows with a vaccine containing killed whole T foetus. Twenty nonpregnant beef cows were randomly assigned to vaccine or control groups as follows: Vaccine (n = 10): cows received 2 mL of a commercial vaccine containing killed whole T foetus subcutaneously and a 2-mL booster 2 weeks later. Control (n = 10): cows received 2 mL of sterile saline on the same schedule. Vaginal secretions and blood samples were collected on Days 0, 8, 15, 22, 29, 36, 43, 50, 60, 75, 89, 110, 146, and 182 relative to day of primary vaccination. Uterine flush fluid was collected on Days 0, 15, 29, and 43 after the day of primary vaccination. Samples were assayed for IgG antibodies to the killed whole T foetus and surface antigen TF1.17 using enzyme-linked immunosorbent assay. Serum whole T foetus-specific IgG levels were significantly increased (between Days 15 and 182) following vaccination with T foetus or with saline. No differences between vaccinates and controls in uterine responses to whole-cell antigen were detected. Serum anti-TF1.17 IgG responses to vaccination were significantly higher than Day 0 throughout the immunization period (P 0.001) and were higher than responses in control animals on each day post immunization through Day 146 (P 0.001). A significant rise in TF1.17-specific IgG levels was observed in vaginal and uterine fluids from Day 15 post vaccination compared to the Day 0 levels. These levels remained significantly elevated in vaginal and uterine fluids through Days 75 (P 0.05) and 43 (P 0.001) after primary vaccination, respectively. Antibody levels in serum, vaginal, and uterine secretions against TF1.17 remained low in the control group throughout the study. In conclusion, vaccination of beef cows with a commercial vaccine containing T foetus induced significant increase in the levels of IgG to the T foetus TF1.17 surface antigen in serum, vaginal secretions, and uterine fluid, which remained elevated through Days 43, 75, and 182 in uterine fluids, vaginal secretions, and serum, respectively. Since purified TF1.17 antigen has been shown to protect against experimental T foetus infection in heifers, the vaccine-induced TF1.17-specific IgG response is likely to be important in the prevention of trichomoniasis in beef cattle.

Trichomoniasis, caused by the protozoan parasite Trichomonas vaginalis , is the most common, non-viral, sexually transmitted infection in the world, but only two closely related nitro drugs are approved for its treatment. New antimicrobials against trichomoniasis remain an urgent need. Several organic gold compounds were tested for activity against T. vaginalis thioredoxin reductase (TrxR) in cell-free systems as well as for activity against different trichomonads in vitro and in a murine infection model. The organic gold(I) compounds auranofin and chloro(diethylphenylphosphine)gold(I) inhibited TrxR in a concentration-dependent manner in assays with recombinant purified reductase and in cytoplasmic extracts of T. vaginalis transfected with a haemagglutinin epitope-tagged form of the reductase. Auranofin potently suppressed the growth of three independent clinical T. vaginalis isolates as well as several strains of another trichomonad ( Tritrichomonas foetus ) in a 24 h-assay, with 50% inhibitory concentrations of 0.7–2.5 µM and minimum lethal concentrations of 2–6 µM. The drug also compromised the ability of the parasite to overcome oxidant stress, supporting the notion that auranofin acts, in part, by inactivating TrxR-dependent antioxidant defences. Chloro(diethylphenylphosphine)gold(I) was 10-fold less effective against T. vaginalis in vitro than auranofin. Oral administration of auranofin for 4 days cleared the parasites in a murine model of vaginal T. foetus infection without displaying any apparent adverse effects. The approved human drug auranofin may be a promising agent as an alternative treatment of trichomoniasis in cases when standard nitro drug therapies have failed.

Bovine trichomoniasis is a concern for the cattle industry. Advances in testing systems have increased the ability to detect the disease in bulls. However, the greatest limitation is proper collection of an adequate sample. The low repeatability observed with most sample collection techniques can cause false-negative results. The aim of our study was to validate a sample collection technique that increases diagnostic sensitivity and is easier and safer to collect than preputial scraping. Commercial bulls (n = 111) of unknown infection status were sampled for detection of Tritrichomonas foetus using 2 different collection methods: 1) preputial scraping with a dry insemination pipette and 2) penile sponging with a 16-ply gauze sponge. Preputial scraping samples were collected by vigorously scraping preputial and penile mucosa using a rigid insemination pipette while applying negative pressure with a syringe. Penile sponge samples were obtained by swabbing the penile and preputial mucosa with a gauze sponge during full extension of the penis. All samples were processed using a commercial medium and submitted under similar conditions for PCR testing. Positive PCR results were detected in 37 of 111 (33%) bulls using the preputial scraping technique; however, 39 of 111 (35%) were positive using the penile sponging technique. The Newton–Raphson algorithm predicted that the sensitivity of the preputial scraping method was 0.919 (95% CI: 0.689–0.983) and the sensitivity of the penile sponging was 0.949 (95% CI: 0.818–0.987). These data indicate that the penile sponging technique is a reliable alternative to the preputial scraping method.

Tritrichomonas foetus is a sexually transmitted protozoan parasite that causes abortions in cattle and results in severe economic losses. In the United States, there are no safe and effective treatments for this parasite and infected animals are typically culled. In order to expedite drug discovery efforts, we investigated in vitro trophozoite killing assays amenable to high-throughput screening in 96 well plate formats. We evaluated the reduction of resorufin, incorporation of propidium iodide, and a luminescence-based ATP detection assay. Of these methods, reduction of resorufin was found to be the most reliable predictor of trophozoite concentrations. We further validated this method by conducting dose-response experiments suitable for calculation of EC50 values for two established compounds with known activity against trophozoites in vitro, namely, metronidazole and ronidazole. Our results demonstrate that the resorufin method is suitable for high-throughput screening and could be used to enhance efforts targeting new treatments for bovine trichomoniasis.