Effects of bacterial contamination of media on the diagnosis of Tritrichomonas foetus by culture and real-time PCR

Clothier, Kristin A.; Villanueva, Michelle; Torain, Andrea; Hult, Cynthia; Wallace, R.

doi:10.1016/j.vetpar.2015.01.006

Cited by 14 publications

(18 citation statements)

References 25 publications

(46 reference statements)

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“…According to the results obtained on the morphology, small differences in the size and shape of the analyzed T. foetus trophozoites and to other morphological studies may be related with the medium used, cultivation time or fixing methods, or even with bacteria present in the medium (Kirby 1951, Tachezy et al 2002, Clothier et al 2015. The trophozoites of T. foetus show consistently three anterior flagella, unlike P. hominis which presents five flagella (Li et al 2014b).…”

Section: Discussionmentioning

confidence: 89%

Co-infection by Tritrichomonas foetus and Pentatrichomonas hominis in asymptomatic cats

et al. 2015

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RESUMO.-[Co-infecção por Tritrichomonas foetus e Pentatrichomonas hominis em gatos assintomáticos.] Tritrichomonas foetus, um parasito bem conhecido por seu significado como um agente patogênico transmitido venereamente em bovinos, também foi identificado como causa de diarreia crónica do intestino grosso em gatos domésticos em muitos países. No Brasil, vários estudos sobre o diagnósti-co de tricomonose bovina foram realizados, mas até agora, não há informação disponível em relação à trichomonose felina. Assim, este é o primeiro estudo a relatar a ocorrência Tritrichomonas foetus, a parasite well known for its significance as a venereally transmitted pathogen in cattle, has been identified as a cause of chronic large bowel diarrhea in domestic cats in many countries of the world. In Brazil, several studies on the diagnosis of bovine trichomoniasis have been performed, but until now, no study was made regarding feline trichomoniasis. Thus, this is the first study to report the occurrence of T. foetus and Pentatrichomonas hominis in cats using morphological and molecular analysis. Feces from 77 cats were examined, four of which (5.2%) were positive for the presence of parabasalids. Morphological analysis of stained smears revealed piriform trophozoites showing the three anterior flagella, elongated nucleus and axostyle ending abruptly in fillet, characteristic of T. foetus. In scanning and transmission electron microscopy, identification characters similar to those previously reported for T. foetus were observed. The cultures containing trophozoites were submitted for molecular analysis, which resulted positive for T. foetus DNA using specific primers (TFR3 and TFR4), and all samples were positive and subjected to sequencing in which they showed 99.7-100% similarity with another isolate sequencing of T. foetus (JX960422). Although no trophozoite with consistent morphology of P. hominis has been visualized in the samples, differential diagnosis was performed using specific primers for P. hominis (TH3 and TH5) amplicon. In three of the four samples (3.89%) sequencing revealed 100% similarity when compared with another sequence of P. hominis deposited in Genbank (KC623939). Therefore, the present study revealed through the diagnostic techniques employed the simultaneous infection by T. foetus and P. hominis in the feces of cats. However, it was necessary to use more than one technique for the diagnosis of the co-infection. These results demonstrate the importance of a correct diagnosis to allow an appropriate treatment by the veterinarian.

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Section: Discussionmentioning

confidence: 89%

Co-infection by Tritrichomonas foetus and Pentatrichomonas hominis in asymptomatic cats

et al. 2015

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“…Fecal components, such as polysaccharide complexes, bile salts, hemoglobin degradation products, phenolic compounds, and heavy metals are generally co-extracted with the pathogen DNA and may interfere with the PCR (GOOKIN et al, 2002;STAUFFER et al, 2008). Presence of bacteria also interfere with T. foetus identification by PCR, as well as in the fecal culture (CLOTHIER et al, 2015).…”

Section: Polymerase Chain Reaction (Pcr)mentioning

confidence: 99%

What is known about Tritrichomonas foetus infection in cats?

Bastos

Mendes-de-Almeida

Brener

2019

Rev. Bras. Parasitol. Vet.

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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.

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“…Representative samples of approximately 30 mg of faeces were placed in the InPouch TM TF-Feline medium, and incubated according to the manufacturer's instructions. Media with bacterial overgrowth or gas production were ruled out from further examination according to the manufacturer's instructions, a necessary step because bacterial overgrowth can interfere with the propagation of trichomonads in the InPouch™ medium (Clothier et al 2015). The faecal flotation method was used concurrently to identify other parasitic infections.…”

Section: Methodsmentioning

confidence: 99%

A survey of feline trichomonosis suggests a low incidence of Tritrichomonas blagburni among cats in the Czech Republic

Ceplecha¹,

Svobodová²,

Lendon³

et al. 2017

Vet. Med. - Czech

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ABSTRACT:Tritrichomonas blagburni (previously called T. foetus) has been implicated as an aetiological agent of long-term large-bowel diarrhoea in cats in many countries worldwide. The aim of this study was to determine the presence of, and risk factors for T. blagburni among a cohort of cats living in different conditions in the Czech Republic. Samples were collected from 170 cats living in different environments. The InPouch TM TF-Feline medium method was used for diagnosis of feline trichomonosis. A single case (0.6%) with motile trichomonads identified as Pentatrichomonas hominis was found in a cat from a multi-cat household. Our study suggests that trichomonads and in particular, T. blagburni, infection may be much less common in the Czech Republic than in neighbouring countries, despite the inclusion of cats that were likely to be from higher-risk groups. A review of studies of the association of trichomonads and feline diarrhoea carried out in different countries revealed variation in the frequency of trichomonads detected. Different combinations of PCR or culture methods for screening or confirmation have been utilised, with or without species differentiation; however, this could not solely account for the variation in the occurrence between countries. From those studies where differentiation was performed, we calculated from the combined studies that T. blagburni occurred in six cats without diarrhoea (1.1%) and 47 cases with diarrhoea (5%). This finding supports an association with diarrhoea as well as the occurrence of asymptomatic cases. We note that in many studies, including our own, the occurrence of T. blagburni may well be underestimated and suggest that future studies use a combination of PCR screening of both faeces and faecal cultures, with differentiation of trichomonad species.

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Effects of bacterial contamination of media on the diagnosis of Tritrichomonas foetus by culture and real-time PCR

Cited by 14 publications

References 25 publications

Co-infection by Tritrichomonas foetus and Pentatrichomonas hominis in asymptomatic cats

Co-infection by Tritrichomonas foetus and Pentatrichomonas hominis in asymptomatic cats

What is known about Tritrichomonas foetus infection in cats?

A survey of feline trichomonosis suggests a low incidence of Tritrichomonas blagburni among cats in the Czech Republic

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