Activation of murine macrophages by membrane proteins from Tritrichomonas foetus grown on iron- and calcium-rich conditions.

Tritrichomonas foetus is a protozoan parasite that causes a venereal disease in cattle limiting reproduction by abortions and sterility. The immune response against this parasite is poorly understood. Since the iron and calcium ions are important regulators of the microenvironment of the urogenital tract in cattle, we decided to evaluate the role of these divalent cations on the antigenicity of membrane proteins of T. foetus on macrophage activation as one of the first inflammatory responses towards this pathogen. Colorimetric methods and ELISA were used to detect the nitric oxide and oxygen peroxide production and expression of cytokines in culture supernatant from macrophage incubated with membrane proteins from T. foetus cultured in iron- and calcium-rich conditions. qRT-PCR assays were used to evaluate the transcript expression of genes involved in the inflammatory response on the macrophages. The membrane proteins used for in vitro stimulation caused the up-regulation of the iNOS and NOX-2 genes as well as the generation of NO and H2 O2 in murine macrophages on a dependent way of the metal concentrations. Additionally, after stimulation, macrophages showed a considerable rise in pro-inflammatory cytokines and a downregulation of anti-inflammatory cytokines, as well as up-regulation in the transcription of the TLR4 and MyD88 genes. These data suggest that membrane proteins of T. foetus induced by iron and calcium can activate an inflammatory specific macrophage response via TLR4/MyD88 signalling pathway.

Ancient and pervasive expansion of adaptin-related vesicle coat machinery across Parabasalia.

The eukaryotic phylum Parabasalia is composed primarily of anaerobic, endobiotic organisms such as the veterinary parasite Tritrichomonas foetus and the human parasite Trichomonas vaginalis, the latter causing the most prevalent, non-viral, sexually transmitted disease world-wide. Although a parasitic lifestyle is generally associated with a reduction in cell biology, T. vaginalis provides a striking counter-example. The 2007 T. vaginalis genome paper reported a massive and selective expansion of encoded proteins involved in vesicle trafficking, particularly those implicated in the late secretory and endocytic systems. Chief amongst these were the hetero-tetrameric adaptor proteins or 'adaptins', with T. vaginalis encoding ∼3.5 times more such proteins than do humans. The provenance of such a complement, and how it relates to the transition from a free-living or endobiotic state to parasitism, remains unclear. In this study, we performed a comprehensive bioinformatic and molecular evolutionary investigation of the heterotetrameric cargo adaptor-derived coats, comparing the molecular complement and evolution of these proteins between T. vaginalis, T. foetus and the available diversity of endobiotic parabasalids. Notably, with the recent discovery of Anaeramoeba spp. as the free-living sister lineage to all parabasalids, we were able to delve back to time points earlier in the lineage's history than ever before. We found that, although T. vaginalis still encodes the most HTAC subunits amongst parabasalids, the duplications giving rise to the complement took place more deeply and at various stages across the lineage. While some duplications appear to have convergently shaped the parasitic lineages, the largest jump is in the transition from free-living to endobiotic lifestyle with both gains and losses shaping the encoded complement. This work details the evolution of a cellular system across an important lineage of parasites and provides insight into the evolutionary dynamics of an example of expansion of protein machinery, counter to the more common trends observed in many parasitic systems.

Detection of Tritrichomonas foetus by RT-rtPCR in pooled bovine preputial washings.

Trichomonosis is a venereal disease of cattle caused by the protozoan Tritrichomonas foetus. T. foetus infection in cattle herds can be economically costly for cattle producers; therefore, testing is important for detection of the agent. Given that bulls are considered to be subclinical carriers of T. foetus, it is important to detect T. foetus infection prior to movement and/or breeding season. We have described previously the development of an updated set of PCR primers and probes that offer increased sensitivity of T. foetus detection in preputial washings collected in PBS by utilizing reverse-transcription real-time PCR (RT-rtPCR) that targets the 5.8S ribosomal RNA of the T. foetus organism. Here, we report improvements in the updated RT-rtPCR reagents as well as the evaluation of testing of pooled preputial washings. We found that up to 5 preputial washings can be pooled, similar to routine testing practices (InPouch culture), without reducing the sensitivity of detection of T. foetus.

Investigation of Toxoplasma gondii, Neospora caninum and Tritrichomonas foetus in abortions of cattle, sheep and goats in Turkey: Analysis by real-time PCR, conventional PCR and histopathological methods.

The aim of this study was to identify Neosopora caninum, Toxoplasma gondii and Tritricomonas foetus in all cattle aborted fetus samples and N. caninum and T. gondii in sheep and goat aborted fetuses sent to Elazig Veterinary Control Institute during two years. Total genomic DNAs were obtained using a commercial kit. Real-time PCR analysis was performed separately for each agent. Conventional PCR was set up for confirmation of positive samples. Then, fetal brain, heart, lung and liver samples were analysed by hematoxylin-eosin (HE) and Avidin-Biotin Complex (ABC) Immunohistochemistry (IHC) methods. Totally, we tested 55 aborted fetus samples. Of these samples, seven (12.7 %) was belonged to goats, 18 (32.7 %) to sheep and 30 (54.5 %) to cattle. T. gondii was detected in six (10.90 %) samples, and four (7.27 %) of them were positive with Real-time PCR, while only one of these four samples was positive for both classical PCR and IHC. N. caninum was determined by at least one of the three tests in 14 (25.45 %) of the samples studied, while 8 (14.54 %) of the positive samples were detected by Real-time PCR, only two of them were also positive with conventional PCR, eight (14.54 %) samples was determined as positive by IHC. Considering T. foetus in the samples, positivity was determined in two (3.63 %) of 55 aborted fetus (both of which were aborted cattle fetus) by Real-time PCR, while only one of them was positive with conventional PCR, while no positivity was detected with the IHC.

Identification of Pentatrichomonas hominis in preputial washes of bulls in Brazil.

The parabasalid Pentatrichomonas hominis is generally considered to represent a symbiotic component of the gastrointestinal microbiota in a wide variety of vertebrate hosts including humans. Nevertheless, a limited number of studies have raised the possibility that it may act as a pathogen of humans, dogs, and pigs and that some human infections may have a zoonotic origin. Data from North America revealed an association between P. hominis and the bovine urogenital tract, principally in bulls and rarely in cows. The importance of this observation is linked to possible interference in the accurate diagnosis of the economically important venereal pathogen Tritrichomonas foetus. The current study employed culture-based and molecular methods to examine the preputial cavities of four breeding bulls, raised in open pasture in southeastern Brazil, for the presence of trichomonads. Motile protozoa were isolated from three of the bulls and were definitively identified as P. hominis based on nucleotide sequencing of polymerase chain reaction (PCR) amplicons derived from the ribosomal RNA operon (ITS1-5.8S rDNA-ITS2) of the parasite. The potential implications of these findings for bovine and human health are discussed.

In-depth comparative analysis of Tritrichomonas foetus transcriptomics reveals novel genes linked with adaptation to feline host.

Tritrichomonas foetus is a flagellated parasite able to infect cattle, cats, and pigs. Despite its prevalence, feline tritrichomonosis has received markedly less attention than venereal infection, and little information about the molecular mechanisms that participate in feline host infection is available. Through a bioinformatics approach, we integrated public transcriptomic data for three T. foetus isolates and explored the differences at transcript level with a focus on pathogenesis and adaptation processes, particularly for the feline isolate. Our analysis revealed higher abundance levels of predicted virulence factors, such as proteases and surface antigens. Additionally, by a comparative and expression analysis of T. foetus genes, we proposed putative virulence factors that could be involved in feline infection. Finally, we identified a great proportion of predicted transcription factors of the MYB protein family and, by a promoter analysis, we revealed that MYB-related proteins could participate in the regulation of gene transcription in T. foetus. In conclusion, this integrated approach is a valuable resource for future studies of host-pathogen interactions and identifying new gene targets for improved feline tritrichomonosis diagnosis and treatment.

Molecular detection of Tritrichomonas foetus in bovine samples: a novel real-time polymerase chain reaction (PCR) assay targeting EF1-alpha-Tf1 and a comparative study of published PCR techniques.

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.

Field detection of Tritrichomonas foetus through loop mediated isothermal amplification with CELDA.

An affordable, portable, and easy-to-build instrument was implemented to perform loop-mediated isothermal amplification (LAMP) in the field. Controlled by Arduino, CELDA heats the reaction tubes, reads the intensity of green fluorescent light and stores data in a computer. In the laboratory and in the field, CELDA was successfully used for the detection of Tritrichomonas foetus DNA in less than 60 min and helped distinguish positives with less difficulty than visual inspection.

Survey of beef bulls in Brazil to assess their role as source of infectious agents related to cow infertility.

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.

Taguchi method for the optimization of three loop mediated isothermal amplification procedures for Tritrichomonas foetus detection.

Taguchi method was used to optimize loop mediated isothermal amplification tests aimed to amplify segments of the elongation factor 1a1 (tf-ef1a1), the 5,8 ribosomal gene (tf-5,8 r) or the beta tubulin 2 (tf-btub2) from the protozoan parasite Tritrichomonas foetus. L9 orthogonal array and quadratic loss functions that penalize deviations from prediction values revealed the effect of amplification reaction components. Analysis of variance (ANOVA) decomposed the contribution of individual factors to a small Ct. Confirmation experiments established that optimum conditions were predictable, verifiable and reproducible. Primers concentration conditioned the non-specific amplification of tf-ef1a1 while betaine and magnesium concentration contributed to accelerate the time to reach a positive threshold in tf-ef1a1, tf-5,8 r and tf-btub2. The general strategy of simple and robust experimental design holds potential as a general optimization protocol for LAMP tests in every diagnostic laboratory.

Tritrichomonas foetus in cats from Central Italy: Clinical signs and risk factors.

Tritrichomonas foetus (Trichomonadida, Tritrichomonadidae) is a parasitic protozoan which causes a diarrheic disease in domestic cats worldwide. The epizootiological and pathogenic impact of this parasite in many geographic locations is still to be unraveled; thus, the present study has investigated the occurrence of T. foetus in cats from central Italy, along with the analysis of risk factors and clinical signs. One hundred and five cats referred to the University Veterinary Teaching Hospital of the University of Teramo were tested with copromicroscopic techniques for common intestinal and extra-intestinal parasites and a specific PCR for T. foetus. A statistical analysis evaluating associations between T. foetus positivity and possible risk factors was performed. One animal scored positive for Giardia (0.95%), 2 for Toxocara cati (1.90%), 1 for Eucoleus aerophilus (syn. Capillaria aerophila) (0.95%), 9 for Aelurostrongylus abstrusus (8.57%) and 1 for Dipylididum caninum (0.95%), while T. foetus was molecularly recorded in 18/105 (17.14%) of the study cats. In two cats, T. foetus was found in association respectively with Giardia and A. abstrusus. Animals with chronic gastrointestinal signs were more significantly infected by T. foetus (40.90% vs 10.84%; p < 0.01; 95% CI = 1.90-17.04; OR = 5.69) in comparison to cats with no gastrointestinal distresses. No other statistically significant associations were found. This study confirms that T. foetus is enzootic in cats in Italy and that it should be included in the differential diagnosis of feline chronic gastrointestinal diseases.

Molecular prevalence of trichomonad species from pet shop puppies and kittens in Japan.

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).

Whole genome sequencing of a feline strain of Tritrichomonas foetus reveals massive genetic differences to bovine and porcine isolates.

Tritrichomonas foetus is a protozoan parasite that colonizes the reproductive tract of cattle as well as the gastrointestinal tract of cats. Bovine tritrichomonosis is a sexually transmitted disease whereas feline tritrichomonosis is thought to be transmitted by the fecal-oral route. Furthermore, T. foetus is known as an essentially apathogenic commensal located in the nasal cavity of pigs. Transmission of T. foetus between the different hosts has to be considered a realistic scenario that may have important implications for the epidemiology of infections and disease. In our study, we generated whole genome sequencing (WGS) data from bovine, feline and porcine T. foetus strains to investigate the genetic (dis)similarities among these diverse strains. As a reference, we used a previously released draft assembly from a bovine T. foetus strain K isolated from an infected bull in Brazil. In particular, we identified single nucleotide polymorphisms (SNPs) and the insertion-deletion (indel) variations within the genomes of the different strains. Interestingly, only a low degree of polymorphism (68 SNPs and indels) was found between the bovine and the porcine strains in terms of variants with a predicted impact of moderate or high and where one species is homozygous for one allele and the other homozygous for the other allele. Conversely, however, a 964 times higher number of such differences was detected by comparing the feline with either the bovine (65,569) or the porcine (65,615) strain. These data clearly indicated a close phylogenetic relationship between bovine and porcine T. foetus but a remarkable genetic distinctness of these two strains from the feline strain. The latter observation was confirmed by PCR-based sequencing of 20 in silico-selected indel markers and five in silico-selected SNP markers that uniformly demonstrated a relatively distant phylogenetic relationship of three independent feline T. foetus isolates in comparison to the bovine and porcine strains investigated. In summary, our comparative genome sequencing approach provided further insights into the genetic diversity of T. foetus in relation to the different host origins of the parasite. Furthermore, our study identified a large number of SNP- and indel-containing sequences that may be useful molecular markers for future epidemiological studies aimed at the elucidation of the transmission patterns of T. foetus within different host species.

Clinical isolates of Tritrichomonas foetus in bulls in Wyoming, South Dakota and Montana, USA.

BACKGROUND: Several Tritrichomonas species have been found in mammalian hosts. Among these trichomonads T. foetus is often found in the urogenital tract of cattle and the gastrointestinal tract of the domestic cat, resulting in sexually transmitted bovine trichomonosis and fecal-orally transmitted feline trichomonosis, respectively. The aims of the current study were to molecularly characterize clinical isolates of T. foetus in cattle populations in Wyoming, South Dakota, and Montana of the United States of America and to phylogenetically analyze Tritrichomonas species of mammalian hosts. RESULTS: DNA sequencing of rRNA genes showed over 99% identity of the newly described isolates to other bovine isolates. Further, T. foetus isolates of various mammalian hosts originated in different geographic regions worldwide were clustered into two well-defined clades by phylogenetic analysis of rRNA and cysteine protease 2 genes. Clade I consisted of isolates originated from cattle, pig, and human whereas clade II contained isolates of cat and dog. CONCLUSION: It is concluded that all mammalian Tritrichomonas spp. apparently belong to T. foetus. Analysis of more sequences is warranted to support this conclusion.

Molecular prevalence of trichomonad species from pet shop puppies and kittens in Japan / Molecular prevalência de espécies trichomonas em filhotes de cães e gatos de pet shop no Japão

Abstract 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).

Multilocus analysis reveals further genetic differences between Tritrichomonas foetus from cats and cattle.

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.

What is known about Tritrichomonas foetus infection in cats?

Tritrichomonas foetus is a parasite that has been definitively identified as an agent of trichomonosis, a disease characterized by chronic diarrhea. T. foetus colonizes portions of the feline large intestine, and manifests as chronic and recurrent diarrhea with mucus and fresh blood, which is often unresponsive to common drugs. Diagnosis of a trichomonad infection is made by either the demonstration of the trophozoite on a direct fecal smear, fecal culture and subsequent microscopic examination of the parasite, or extraction of DNA in feces and amplification by the use of molecular tools. T. foetus is commonly misidentified as other flagellate protozoa such as Giardia duodenalis and Pentatrichomonas hominis. Without proper treatment, the diarrhea may resolve spontaneously in months to years, but cats can remain carriers of the parasite. This paper intends to serve as a source of information for investigators and veterinarians, reviewing the most important aspects of feline trichomonosis, such as trichomonad history, biology, clinical manifestations, pathogenesis, world distribution, risk factors, diagnosis, and treatment.

Fulminant Tritrichomonas foetus 'feline genotype' infection in a 3-month old kitten associated with viral co-infection.

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.

What is known about Tritrichomonas foetus infection in cats? / O que sabemos sobre a infecção por Tritrichomonas foetus em gatos?

Abstract Tritrichomonas foetus is a parasite that has been definitively identified as an agent of trichomonosis, a disease characterized by chronic diarrhea. T. foetus colonizes portions of the feline large intestine, and manifests as chronic and recurrent diarrhea with mucus and fresh blood, which is often unresponsive to common drugs. Diagnosis of a trichomonad infection is made by either the demonstration of the trophozoite on a direct fecal smear, fecal culture and subsequent microscopic examination of the parasite, or extraction of DNA in feces and amplification by the use of molecular tools. T. foetus is commonly misidentified as other flagellate protozoa such as Giardia duodenalis and Pentatrichomonas hominis. Without proper treatment, the diarrhea may resolve spontaneously in months to years, but cats can remain carriers of the parasite. This paper intends to serve as a source of information for investigators and veterinarians, reviewing the most important aspects of feline trichomonosis, such as trichomonad history, biology, clinical manifestations, pathogenesis, world distribution, risk factors, diagnosis, and treatment.

Resumo Tritrichomonas foetus é um parasito que foi identificado definitivamente como agente de tricomoníase, caracterizada por diarreia crônica. T. foetus coloniza porções do intestino grosso dos felinos e se manifesta como uma diarreia crônica e recorrente, com muco e sangue, geralmente irresponsiva às drogas comumente usadas no tratamento. O diagnóstico da infecção por tricomonadídeos é feito pela demonstração de trofozoítos no exame direto de fezes frescas, cultura fecal e subsequente exame microscópico ou extração do DNA do parasito na amostra fecal e amplificação, utilizando-se técnicas moleculares. T. foetus é comumente confundido com outros protozoários flagelados, como Giardia duodenalis e Pentatrichomonas hominis. Sem tratamento adequado, a diarreia pode cessar espontaneamente em meses ou anos, porém os gatos podem permanecer portadores do parasito. Esse artigo pretende servir como fonte de informação para pesquisadores e veterinários, revisando os mais importantes aspectos da tricomoníase felina, como histórico, biologia, manifestações clínicas, patogênese, distribuição mundial, fatores de risco, diagnóstico e tratamento.

Comparison of multiplexed-tandem real-time PCR panel with reference real-time PCR molecular diagnostic assays for detection of Giardia intestinalis and Tritrichomonas foetus in cats.

Giardia intestinalis and Tritrichomonas foetus are frequent enteric protozoan parasites of the gastrointestinal track of domestic cats. Because of different treatment options for the parasites, confirmation of presence of one or both pathogens is necessary. The PCR based assays are suitable for differential diagnosis. We evaluated the performance of Small Animal Diarrhoea panel, a multiplexed-tandem real-time PCR (MT-PCR) assay, that detects DNA of both G. intestinalis and T. foetus. The sensitivity and specificity were compared to reference real-time PCR assays using 105 faecal samples, 39.05% (n = 41) positive for G. intestinalis and 30.48% (n = 32) were positive for T. foetus. The faecal samples positive for T. foetus had a high proportion of late amplifiers, determined by an arbitrary threshold of Ct-values > 35. On the other hand, only one G. intestinalis positive sample was considered a late amplifier. For G. intestinalis DNA, the MT-PCR assay had 95.1% sensitivity and 92.1% specificity. For T. foetus DNA, the MT-PCR assay had 41.9% sensitivity and 100.0% specificity. To evaluate the interlaboratory reproducibility of the MT-PCR assay, results were compared in two different laboratories and found to be in a very good agreement (Kappa = 0.9). Further analysis of the DNA using conventional PCR determined presence of G. intestinalis Assemblage F and T. foetus genotype 'feline'. In conclusion, the MT-PCR Small Animal Diarrhoea panel had a good and poor performance against reference assays for G. intestinalis and T. foetus, respectively. The assay is suitable for detection and differential diagnosis of G. intestinalis and moderate to high burdens of T. foetus in small animal clinical practice.