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Tritrichomonas foetus is a protozoan parasite that has been recently identified as a causative agent of chronic diarrhea in domestic cats. Transmission of infection occurs by the fecal-oral route through direct contact among animals. Consequently, feline trichomonosis (FT) is more likely to be present in multi-cat environments. The objective of this work was to study the presence of T. foetus and some associated risk factors in cats from densely housed origins and with a reported history of chronic diarrhea. Animals enrolled in this study were family cats (n = 15) acquired from pet shops, shelters or breeding centers and cattery cats belonging to one breeding center (n = 28) and two cat shelters (A and B, n = 25 each). In the catteries, a follow-up analysis for a period of up to 2 months was also performed to determine the parasite shedding pattern in feces and the incidence of infection. Fecal samples were analyzed using in vitro culture and a PCR technique. T. foetus was detected in a total of 38.7% (36/93) of the cats with chronic diarrhea. Parasite infection was similarly detected in family cats and cattery animals (40% versus 38.4%). In the catteries, the parasite was detected in 50%, 44% and 20% of the animals from the breeding center and shelters A and B, respectively. The follow-up analysis showed that 58.3% of infected cats intermittently shed trophozoites in their feces, with an incidence of 23.1%. Investigation of potential risk factors showed that cats ≤1 year old were more likely to be infected than older cats (57.1% versus 27.3%; P < 0.05). No significant differences were found when sex and breed factors were studied. These results confirm the importance of FT as a cause of chronic diarrhea in cats and highlight the relevance of close contact conditions for T. foetus transmission., Depto. de Sanidad Animal, CAI Ciencias de la Tierra y Arqueometría, TRUE, pub

Tritrichomonas foetus causes a venereal disease in cattle called bovine trichomoniasis. T. foetus lives in the reproductive tract of the cow, the surface of the penis and prepuce of the bull and causing early fetal loss and sometimes late term abortions. It can also extend breeding / calving season. In this study, 155 cows of different ages (2-6 years old) and with a variety of clinical signs (repeat breeders, abortions, and discharge only) were examined from different areas in Basrah Province, Iraq from November 2020 to July 2021, and 75 samples collected from healthy animals (without any symptom). The infection rate was detected in wet mount techniques found fewer positive findings than in pouch techniques, with 12 (40%) recorded and in pouch systems, 30 (100%) recorded while in the culture 24(80 %), Giemsa-stained smears 19(63.3%) and Acridine orange was 20 (66.6%). The present study improved that cows more than three years are more susceptible to be infected than cows less than three years. The study concluded that if no control method is applied in Basrah, there is a serious risk of spreading Trichomonas foetus.

Tritrichomonas foetus causes a venereal disease in cattle called bovine trichomoniasis. T. foetus lives in the reproductive tract of the cow, the surface of the penis and prepuce of the bull and causing early fetal loss and sometimes late term abortions. It can also extend breeding / calving season. In this study, 155 cows of different ages (2-6 years old) and with a variety of clinical signs (repeat breeders, abortions, and discharge only) were examined from different areas in Basrah Province, Iraq from November 2020 to July 2021, and 75 samples collected from healthy animals (without any symptom). The infection rate was detected in wet mount techniques found fewer positive findings than in pouch techniques, with 12 (40%) recorded and in pouch systems, 30 (100%) recorded while in the culture 24(80 %), Giemsa-stained smears 19(63.3%) and Acridine orange was 20 (66.6%). The present study improved that cows more than three years are more susceptible to be infected than cows less than three years. The study concluded that if no control method is applied in Basrah, there is a serious risk of spreading Trichomonas foetus.

Tritrichomonas foetus causes a venereal disease in cattle called bovine trichomoniasis. T. foetus lives in the reproductive tract of the cow, the surface of the penis and prepuce of the bull and causing early fetal loss and sometimes late term abortions. It can also extend breeding / calving season. In this study, 155 cows of different ages (2-6 years old) and with a variety of clinical signs (repeat breeders, abortions, and discharge only) were examined from different areas in Basrah Province, Iraq from November 2020 to July 2021, and 75 samples collected from healthy animals (without any symptom). The infection rate was detected in wet mount techniques found fewer positive findings than in pouch techniques, with 12 (40%) recorded and in pouch systems, 30 (100%) recorded while in the culture 24(80 %), Giemsa-stained smears 19(63.3%) and Acridine orange was 20 (66.6%). The present study improved that cows more than three years are more susceptible to be infected than cows less than three years. The study concluded that if no control method is applied in Basrah, there is a serious risk of spreading Trichomonas foetus.

Tritrichomonas foetus isolates from feline and bovine origin has been previously shown to carry a certain degree of genetic heterogeneity. Here, novel candidate molecular markers were developed by means of multilocus sequence typing of the gap2 gene (encoding for T. foetus glyceraldehyde-3-phosphate dehydrogenase), ITS region, the TR7/TR8 variable-length repeat and microsatellite genotyping. These markers were used to characterize T. foetus field isolates from bulls and domestic cats and to compare phylogenetically with the following ATCC isolates: T. foetus isolated from cattle and pig (syn. Tritrichomonas suis), Tritrichomonas mobilensis, Tetratrichomonas gallinarum and Pentatrichomonas hominis. Among them, TFMS10 and TFMS7 were found to be the most polymorphic markers. Moreover, an 809 bp fragment of the gap2 gene was successfully amplified from all the trichomonads included in this study and the sequence analysis revealed differences between T. foetus porcine and feline genotypes and T. mobilensis in comparison to the bovine T. foetus ATCC isolate. The TR7/TR8 repeat pattern was not reproducible, being only consistent the fragments of approximately 110 and 217 bp. Sequence analysis of the latter revealed the existence of 3 SNPs resulting in 98.6 % homology between bovine and feline isolates. A search for similar sequences was carried out to develop a Restriction Length Fragment Polymorphism analysis. A 503 bp region, named TF1, revealed the existence of two BbvI restriction enzyme sites that were able to generate different length fragments for T. foetus feline and bovine isolates. Finally, the neighbour-joining analyses showed that T. foetus porcine genotype clusters together with bovine genotype, whereas T. mobilensis and the feline genotype form a separate cluster., Depto. de Sanidad Animal, Fac. de Veterinaria, TRUE, pub

CRUE-CSIC (Acuerdos Transformativos 2022), The parasite T. foetus causes trichomonosis in cattle but is generally asymptomatic in males. Thus, many bulls carrying the disease go unnoticed, making the detection of T. foetus in bulls an important aspect for its control. Due to drawbacks posed by its cultivation, PCR is a preferred option for diagnostic laboratories. Most published PCR protocols target the genomic region compring the 18S, 5.8S, and 28S rRNA genes and internal transcribed spacers 1 and 2 (rRNA-ITS region), homologous to that of other Tritrichomonas species. There is minimal information on alternative genetic targets and no comparative studies have been published. We compared a protocol based on the microsatellite TfRE (called H94) and five protocols based on the rRNA-ITS region (called M06, M15, G02, G05, and N02). We also designed and evaluated a novel PCR-based assay on the EF1-alpha-Tf1 gene (called V21). The analytical sensitivity and specificity assays for the PCR protocols were performed according to the World Organisation for Animal Health (OIE) directives and the comparative study was performed with a widely used PCR (M06) on clinical samples from 466 breeding bulls. V21 showed a high degree of agreement with our reference M06 (kappa = 0.967), as well as M15 (kappa = 0.958), G05 (kappa = 0.948), and H94 (kappa = 0.986). Protocols H94 and V21 appear to be good approaches for confirming clinical cases in preputial bull samples when genomic regions alternative to rRNA-ITS are required. By contrast, N02 gave false negatives and G02 false positives., Comunidad de Madrid, R&D Agreement between the Instituto Tecnológico Agrario de Castilla y León, VISAVET-Universidad Complutense de Madrid, and Universidad de Burgos, Depto. de Sanidad Animal, Fac. de Veterinaria, Centro de Vigilancia Sanitaria Veterinaria (VISAVET), TRUE, pub

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.

Tritrichomonas foetus is a sexually transmitted protozoon that causes genital inflammation and adverse pregnancy outcomes in cattle. Cysteine proteinases (CPs) released by T. foetus degrade immunoglobulin G (IgG) antibodies, complement component 3 and matrix proteins as well as inducing apoptosis of bovine genital epithelial cells. In this study, the efficacies of the vinyl sulfone CP inhibitors K11777 and WRR-483 were tested against CPs of T. foetus. The activity of secreted T. foetus CPs in culture supernatants was decreased in the presence of vinyl sulfone inhibitors. Inhibitor K11777 reduced the in vitro cytopathogenic effects of T. foetus in bovine foetal trophoblast cells, which are relevant target cells since this pathogen interferes with pregnancy. Pre-treatment of T. foetus prior to intravaginal inoculation diminished genital infection in a murine model. Therefore, vinyl sulfone CP inhibitors reduce several effects of T. foetus-secreted CPs, including cytotoxicity on relevant target host cells and genital infection in a murine model. These inhibitors have potential as chemotherapeutic agents against bovine trichomoniasis. Generalisation to human trichomoniasis requires further study.

Tritrichomonas foetus is a sexually transmitted protozoon that causes genital inflammation and adverse pregnancy outcomes in cattle. Cysteine proteinases (CPs) released by T. foetus degrade immunoglobulin G (IgG) antibodies, complement component 3 and matrix proteins as well as inducing apoptosis of bovine genital epithelial cells. In this study, the efficacies of the vinyl sulfone CP inhibitors K11777 and WRR-483 were tested against CPs of T. foetus. The activity of secreted T. foetus CPs in culture supernatants was decreased in the presence of vinyl sulfone inhibitors. Inhibitor K11777 reduced the in vitro cytopathogenic effects of T. foetus in bovine foetal trophoblast cells, which are relevant target cells since this pathogen interferes with pregnancy. Pre-treatment of T. foetus prior to intravaginal inoculation diminished genital infection in a murine model. Therefore, vinyl sulfone CP inhibitors reduce several effects of T. foetus-secreted CPs, including cytotoxicity on relevant target host cells and genital infection in a murine model. These inhibitors have potential as chemotherapeutic agents against bovine trichomoniasis. Generalisation to human trichomoniasis requires further study.

Tritrichomonas foetus es un protozoo flagelado que, cuando parasita el colon del gato, puede causar diarrea crónica. Causante de patología reproductiva en bovinos, la T. foetus aislada en el gato posee un genotipo distinto al bovino. Suele afectar a gatitos que generalmente viven en colectividades como criaderos, sin predisposición de género ni raza, o también a colonias de gatos mestizos. El signo clínico predominante es la diarrea de intestino grueso crónica o intermitente. Las heces varían de pastosas a semilíquidas, con presencia de moco y/o sangre. El diagnóstico se basa, bien en la visualización del parásito en examen directo de heces (de muestras no más antiguas de 6 horas), o bien mediante medios de cultivo específicos o la realización de PCR de RNA ribosomal. La histopatología muestra colitis linfoplasmocíticas y/o neutrofílicas. Una vez tratadas las frecuentes coinfecciones,es frecuente que la diarrea se resuelva de forma espontánea tras cierto tiempo (media de 9 meses). El único tratamiento médico efectivo para la trichomonosis es el ronidazol, siendo su efecto adverso más importante la neurotoxicidad.

A cytosolic malate dehydrogenase from Tritrichomonas foetus (Riedmüller) was purified to apparent homogeneity by a combination of differential centrifugation, affinity, hydrophobic interaction and ion-exchange chromatography. The purest preparation had specific activity of 309 mumoles.(mg protein)-1.min-1, which corresponded to 50-fold purification. The enzyme representing almost 2% of total cytosolic protein displayed hyperbolic affinity curves for substrate and coenzyme. Km for oxaloacetate and NADH was 17.5 microM and 13 microM, respectively. Subunit size determined by SDS-PAGE and by laser desorption mass spectroscopy was approximately 37 kDa, which corresponds to values reported for most malate dehydrogenases. The native molecular weight determined by gel filtration was 244 kDa, indicating six subunit structure of holoenzyme; this is unusual in comparison with other malate dehydrogenases, which are usually dimers or tetramers.

Tritrichomonas foetus was found to contain NADP-specific malic enzyme. The activity was present in the cytosolic fraction and was about 5-fold higher in extracts of a metronidazole-resistant strain (KV1-1MR-100) than of the parent strain (KVc1). Electrophoresis under non-denaturing conditions and activity staining indicated the existence of 3 isozymes termed I, II and III in order of increasing electrophoretic mobility. Isozymes I and II were much less active than isozyme III in the parent strain, whereas all three isozymes had comparable activities in the resistant strain. NADP-malic enzymes were purified from the cytosolic fraction of the resistant strain to apparent homogeneity and were identified by SDS-PAGE as polypeptides of 41.5 kDa (I), 40.5 kDa (III) and as a mixture of both in equal amounts (II). The molecular mass of the three holoenzymes was about 180 kDa, as determined by gel-filtration on Sephacryl S-300 HR, indicating a tetrameric structure. Isozyme III was also purified from parent strain and shown to consist of the 40.5-kDa polypeptide. Km values for malate were 0.31, 0.65 and 1.35 mM for isozyme I, II and III, respectively. From these results we conclude that T. foetus+, which is required for the formation of ethanol by alcohol dehydrogenase, an NADP-specific enzyme in this species. This is particularly important for the resistant strain, in which ethanol is the major end-product of glucose metabolism.