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Journal of the American Veterinary... May 2003To evaluate the efficacy of and optimize a commercially available culture system for sensitive and specific in-clinic culture of Tritrichomonas foetus from cat feces.
OBJECTIVE
To evaluate the efficacy of and optimize a commercially available culture system for sensitive and specific in-clinic culture of Tritrichomonas foetus from cat feces.
DESIGN
Prospective study.
SAMPLE POPULATION
Samples of freshly voided feces from 117 purebred cats and pure cultures of T. foetus obtained from a cat with chronic diarrhea.
PROCEDURE
Optimal conditions for use of the culture system, such as quantity of fecal inoculum (0.025 to 0.2 g) and cultivation temperature (25 or 37 degrees C [98.6 or 77.0 degrees F]), were determined. Specificity of the system was examined by attempted culture of Giardia lamblia and Pentatrichomonas hominis. Sensitivity of the system to detect T. foetus was determined by inoculation of culture system pouches with serially diluted T. foetus suspensions with and without feces.
RESULTS
Detection limit of the culture system was 1 and 1,000 T. foetus organisms without and with feces from cats, respectively. Optimal fecal inoculum was < 0.1 g of feces. At 37 degrees C, cultures yielded positive results in 24 hours; organisms remained viable for 1 to 6 days, and bacterial overgrowth was common. At 25 degrees C, cultures yielded positive results in 1 to 11 days; organisms were long-lived, and bacterial overgrowth was uncommon. Neither G. lamblia or P. hominis survived in the culture system.
CONCLUSIONS AND CLINICAL RELEVANCE
The culture system was sensitive and specific for culture of T. foetus in feces of cats. Performance was optimal when test kits were inoculated with < or = 0.1 g of freshly voided feces and cultured at 25 degrees C.
Topics: Animals; Cat Diseases; Cats; Feces; Female; Prospective Studies; Protozoan Infections; Protozoan Infections, Animal; Sensitivity and Specificity; Temperature; Time Factors; Tritrichomonas foetus
PubMed: 12762381
DOI: 10.2460/javma.2003.222.1376 -
The Journal of Parasitology Feb 2003Recently, several investigators have reported large-bowel diarrhea in cats associated with intestinal trichomonad parasites. These reports have presumptively identified...
Recently, several investigators have reported large-bowel diarrhea in cats associated with intestinal trichomonad parasites. These reports have presumptively identified the flagellates as Pentatrichomonas hominis, a n organism putatively capable of infecting the intestinal tracts of a number of mammalian hosts, including cats, dogs, and man. The purpose of the present study was to determine the identity of this recently recognized flagellate by means of rRNA gene sequence analysis; restriction enzyme digest mapping; and light, transmission, and scanning electron microscopy (SEM).
Topics: Animals; Base Sequence; Cat Diseases; Cats; DNA, Protozoan; DNA, Ribosomal; Diagnosis, Differential; Diarrhea; Flagella; Microscopy, Electron, Scanning; Molecular Sequence Data; Polymorphism, Restriction Fragment Length; Protozoan Infections; Protozoan Infections, Animal; RNA, Protozoan; RNA, Ribosomal; Sequence Alignment; Trichomonadida; Tritrichomonas foetus
PubMed: 12659310
DOI: 10.1645/0022-3395(2003)089[0099:TFANPH]2.0.CO;2 -
Journal of Veterinary Diagnostic... Jan 2003Sequence analysis of the 5.8S rRNA gene and the internal transcribed spacer regions (ITSRs) was used to compare trichomonadid protozoa (n = 39) of varying morphologies... (Comparative Study)
Comparative Study
Sequence analysis of the 5.8S rRNA gene and the internal transcribed spacer regions (ITSRs) was used to compare trichomonadid protozoa (n = 39) of varying morphologies isolated from the bovine preputial cavity. A multiple sequence alignment was performed with bovine isolate sequences and other trichomonadid protozoa sequences available in GenBank. As a group, Tritrichomonasfoetus isolates (n = 7) had nearly complete homology. A similarity matrix showed low homology between the T. foetus isolates and other trichomonads recovered from cattle (<70%). Two clusters of trichomonads other than T. foetus were identified. Eighteen isolates comprised 1 group. These isolates shared >99% homology among themselves and with Pentatrichomonas hominis. The other non-T. foetus cluster (n = 14) did not exhibit a high degree of homology (<87%) with other bovine isolates or any of the trichomonad sequences available in GenBank. The sequence homology among isolates in that cluster was >99%, except for 1 isolate that varied from the others in both ITSRs (approximately 2% dissimilarity). Sequence analysis of the 5.8S rRNA gene and ITSRs was useful for comparing trichomonadid protozoa isolated from the bovine preputial cavity and demonstrated that 2 distinct types of trichomonads constituted the non-T. foetus isolates recovered from the bovine preputial cavity.
Topics: Animals; Cattle; Cattle Diseases; DNA, Protozoan; DNA, Ribosomal Spacer; Genetic Variation; Male; Molecular Sequence Data; RNA, Ribosomal, 5.8S; Sequence Homology, Nucleic Acid; Sexually Transmitted Diseases; Trichomonas; Trichomonas Infections
PubMed: 12580289
DOI: 10.1177/104063870301500104 -
Journal of Clinical Microbiology Nov 2002Tritrichomonas foetus, a venereal pathogen of cattle, was recently identified as an inhabitant of the large intestine in young domestic cats with chronic diarrhea....
Tritrichomonas foetus, a venereal pathogen of cattle, was recently identified as an inhabitant of the large intestine in young domestic cats with chronic diarrhea. Recognition of the infection in cats has been mired by unfamiliarity with T. foetus in cats as well as misdiagnosis of the organisms as Pentatrichomonas hominis or Giardia sp. when visualized by light microscopy. The diagnosis of T. foetus presently depends on the demonstration of live organisms by direct microscopic examination of fresh feces or by fecal culturing. As T. foetus organisms are fastidious and fragile, routine flotation techniques and delayed examination and refrigeration of feces are anticipated to preclude the diagnosis in numerous cases. The objective of this study was to develop a sensitive and specific PCR test for the diagnosis of feline T. foetus infection. A single-tube nested PCR was designed and optimized for the detection of T. foetus in feline feces by using a combination of novel (TFITS-F and TFITS-R) and previously described (TFR3 and TFR4) primers. The PCR is based on the amplification of a conserved portion of the T. foetus internal transcribed spacer (ITS) region (ITS1 and ITS2) and the 5.8S rRNA gene. The absolute detection limit of the single-tube nested PCR was 1 organism, while the practical detection limit was 10 organisms per 200 mg of feces. Specificity was examined by using P. hominis, Giardia lamblia, and feline genomic DNA. Our results demonstrate that the single-tube nested PCR is ideally suited for (i) diagnostic testing of feline fecal samples that are found negative by direct microscopy and culturing and (ii) definitive identification of microscopically observable or cultivated organisms.
Topics: Animals; Cat Diseases; Cats; DNA Primers; DNA, Protozoan; DNA, Ribosomal; Feces; Polymerase Chain Reaction; Protozoan Infections; Protozoan Infections, Animal; RNA, Ribosomal, 5.8S; Sensitivity and Specificity; Tritrichomonas foetus
PubMed: 12409385
DOI: 10.1128/JCM.40.11.4126-4130.2002 -
Molecular Biology and Evolution Aug 2001Class II fumarase sequences were obtained by polymerase chain reaction from five trichomonad species. All residues known to be highly conserved in this enzyme were...
Class II fumarase sequences were obtained by polymerase chain reaction from five trichomonad species. All residues known to be highly conserved in this enzyme were present. Nuclear run-on assays showed that one of the two genes identified in Tritrichomonas foetus was expressed, whereas no fumarase transcripts were detected in the related species Trichomonas vaginalis. These findings corroborate previous biochemical data. Fumarase genes were also expressed in Monocercomonas sp. and Tetratrichomonas gallinarum but not in Pentatrichomonas hominis, Trichomonas gallinae, Trichomonas tenax, and Trichomitus batrachorum under the culture conditions used. Molecular trees inferred by likelihood methods reveal that trichomonad sequences have no affinity to described class II fumarase genes from other eukaryotes. The absence of functional mitochondria in protists such as trichomonads suggests that they diverged from other eukaryotes prior to the alpha-proteobacterial symbiosis that led to mitochondria. Furthermore, they are basal to other eukaryotes in rRNA analyses. However, support for the early-branching status of trichomonads and other amitochondriate protists based on phylogenetic analyses of multiple data sets has been equivocal. Although the presence of hydrogenosomes suggests that trichomonads once had mitochondria, their class II iron-independent fumarase sequences differ markedly from those of other mitochondriate eukaryotes. All of the class II fumarase genes described from other eukaryotes are of apparent alpha-proteobacterial origin and hence a marker of mitochondrial evolution. In contrast, the class II fumarase from trichomonads emerges among other eubacterial homologs. This is intriguing evidence for an independent acquisition of these genes in trichomonads apart from the mitochondrial endosymbiosis event that gave rise to the form present in other eukaryotes. The ancestral trichomonad class II fumarase may represent a prokaryotic form that was replaced in other eukaryotes after the divergence of trichomonads with the movement of endosymbiont genes into the nucleus. Alternatively, it may have been acquired via a separate endosymbiotic event or lateral gene transfer.
Topics: Amino Acid Sequence; Animals; DNA, Protozoan; Evolution, Molecular; Fumarate Hydratase; Gene Expression Regulation, Enzymologic; Molecular Sequence Data; Phylogeny; RNA, Protozoan; Sequence Alignment; Sequence Analysis, DNA; Sequence Homology, Amino Acid; Trichomonadida
PubMed: 11470849
DOI: 10.1093/oxfordjournals.molbev.a003944 -
Transactions of the Royal Society of... 2000
Topics: Adult; Empyema, Pleural; Fatal Outcome; Female; Humans; Lupus Erythematosus, Systemic; Sequence Alignment; Sequence Analysis, DNA; Trichomonas Infections
PubMed: 10897364
DOI: 10.1016/s0035-9203(00)90270-0 -
Journal of the American Veterinary... Apr 2000Four purebred domestic cats examined because of diarrhea were found to have Pentatrichomonas hominis, a rarely reported trichomonad parasite, in their feces. Treatment...
Four purebred domestic cats examined because of diarrhea were found to have Pentatrichomonas hominis, a rarely reported trichomonad parasite, in their feces. Treatment with a combination of metronidazole and enrofloxacin tended to improve consistency of the feces, whereas treatment with metronidazole alone reduced the number of P hominis trophozoites in fecal smears but did not necessarily result in an improvement in clinical signs. Two cats were euthanatized. Necropsy revealed lymphoplasmacytic enterocolitis with eosinophils and eosinophilic globular leukocytes, neutrophils in the mucosa of the colon and within intraluminal contents of the cecum, and P hominis trophozoites in intraluminal contents of the colon and cecum.
Topics: Animals; Anti-Infective Agents; Antitrichomonal Agents; Cat Diseases; Cats; Colon; Diarrhea; Enrofloxacin; Feces; Female; Fluoroquinolones; Male; Metronidazole; Protozoan Infections; Protozoan Infections, Animal; Quinolones; Trichomonadida
PubMed: 10767968
DOI: 10.2460/javma.2000.216.1270 -
The Journal of Eukaryotic Microbiology 2000We determined small subunit ribosomal DNA sequences from three parabasalid species, Trichomitus batrachorum strain R105, Tetratrichomonas gallinarum, and...
We determined small subunit ribosomal DNA sequences from three parabasalid species, Trichomitus batrachorum strain R105, Tetratrichomonas gallinarum, and Pentatrichomonas hominis belonging to the Trichomonadinae subfamily. Unrooted molecular phylogenetic trees inferred by distance, parsimony, and likelihood methods reveal four discrete clades among the parabasalids. The Trichomonadinae form a robust monophyletic group. Within this subfamily T. gallinarum is closely related to Trichomonas species as supported by morphological data, with P. hominis and Pseudotrypanosoma giganteum occupying basal positions. Our analysis does not place T. batrachorum within the Trichomonadinae. Trichomitus batrachorum (strains R105 and BUB) and Hypotrichomonas acosta form a well-separated cluster, suggesting the genus Trichomitus is polyphyletic. The emergence of T. batrachorum precedes the Trichomonadinae-Tritrichomonadinae dichotomy, emphasizing its pivotal evolutionary position among the Trichomonadidae. A third cluster unites the Devescovinidae and the Calonymphidae. The fourth clade contains the three hypermastigid sequences from the genus Trichonympha, which exhibit the earliest emergence among the parabasalids. The addition of these three new parabasalid species did not however resolve ambiguities regarding the relative branching order of the parabasalid clades. The phylogenetic positions of Tritrichomonas faetus, Monocercomonas sp., Dientamoeba fragilis, and the unidentified Reticulitermes flavipes gut symbiont 1 remain unclear.
Topics: Animals; Cloning, Molecular; DNA, Protozoan; DNA, Ribosomal; Evolution, Molecular; Phylogeny; RNA, Ribosomal; Sequence Analysis, DNA; Trichomonadida
PubMed: 10651299
DOI: 10.1111/j.1550-7408.2000.tb00013.x -
Parasitology Aug 1997The taxonomic situation in the genus Tritrichomonas is the subject of controversial discussion: potentially T. foetus and T. suis, the tritrichomonads from cattle and... (Comparative Study)
Comparative Study
The taxonomic situation in the genus Tritrichomonas is the subject of controversial discussion: potentially T. foetus and T. suis, the tritrichomonads from cattle and swine, respectively, could belong to the same species. In order to shed some light on this question, a molecular biological analysis was performed. The 5.8S rRNA gene and the flanking internal transcribed spacer regions (ITS1 and ITS2) of 12 different isolates of 3 Tritrichomonas species T. foetus, T. suis and T. mobilensis were enzymatically amplified by PCR and subcloned. Also, the corresponding regions of the trichomonads Trichomonas vaginalis, T. tenax, T. gallinae and Pentatrichomonas hominis were included in this study. Sequence analysis of cloned fragments was used to compare the parasite isolates. The genus Tritrichomonas exhibited an extremely high degree of homogeneity. All T. foetus and T. suis isolates had identical sequences, and only 1 substitution was found in the ITS2 region of T. mobilensis. In contrast, the genus Trichomonas shared more diversity. The results obtained in this study support a possible future revision of the taxonomic classification of tritrichomonads.
Topics: Animals; Base Sequence; Cattle; Cattle Diseases; DNA, Protozoan; DNA, Ribosomal; Genes, rRNA; Humans; Molecular Sequence Data; Polymerase Chain Reaction; Protozoan Infections; Protozoan Infections, Animal; RNA, Ribosomal, 5.8S; Sequence Analysis, DNA; Swine; Swine Diseases; Trichomonadida; Trichomonas; Trichomonas Infections; Tritrichomonas
PubMed: 10190167
DOI: 10.1017/s0031182097001212 -
Annals of Tropical Medicine and... Jan 1997To shorten the time and improve the accuracy of diagnosis of vaginal trichomoniasis, a novel, one-tube, nested PCR, which targets a family of 650-bp specific DNA repeats...
To shorten the time and improve the accuracy of diagnosis of vaginal trichomoniasis, a novel, one-tube, nested PCR, which targets a family of 650-bp specific DNA repeats from the Trichomonas vaginalis genome, has been developed. Samples were prepared by a rapid boiling method and the PCR products analysed by gel electrophoresis. A 290-bp DNA fragment was observed in all positive cases. No cross-reaction with any other pathogens, including the Pentatrichomonas hominis and Giardia lamblia used as controls, was found. Using the assay, one genome-equivalent of T. vaginalis in 20 microliters vaginal discharge can be detected and diagnosis can be made within 6 h. When 165 clinical specimens were examined by wet amount, culture and the PCR assay, 16 were found positive for T. vaginalis by both culture and PCR, whereas only nine of these 16 cases were found to be positive by examination of wet mounts. No PCR-negative cases were positive by wet mount or culture. This new assay appears to be a simple, rapid, accurate and sensitive method for the diagnosis of vaginal trichomoniasis.
Topics: Animals; Female; Humans; Microbiological Techniques; Polymerase Chain Reaction; Sensitivity and Specificity; Trichomonas Vaginitis; Trichomonas vaginalis; Vaginal Discharge
PubMed: 9093430
DOI: 10.1080/00034983.1997.11813112