Evidence of self-resolution of feline trichomonosis in a pair of single household cats due to ronidazole-resistant Tritrichomonas foetus.

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.

Repurposing the Veterinary Antiprotozoal Drug Ronidazole for the Treatment of Clostridioides difficile Infection.

Clostridioides difficile infection (CDI) is a principal cause of hospital-acquired infections and fatalities worldwide. The need for new, more potent anticlostridial agents is far from being met. Drug repurposing can be utilized as a rapid and cost-efficient method of drug development. The current study was conducted to evaluate the activity of ronidazole, a veterinary antiprotozoal drug, as a potential treatment for CDI. Ronidazole inhibited the growth of clinical C. difficile isolates (including NAP1 and toxigenic strains) at a very low concentration (0.125 µg/mL) and showed superior killing kinetics compared with metronidazole, an anticlostridial agent from the same chemical category. In addition, ronidazole did not inhibit growth of several commensal organisms naturally present in the human intestine that play a protective role in preventing CDIs. Furthermore, ronidazole was found to be non-toxic to human gut cells and permeated a monolayer of colonic epithelial cells (Caco-2) at a slower rate than metronidazole. Finally, ronidazole outperformed metronidazole when both were tested at a dose of 1 mg/kg daily in a mouse model of CDI. Overall, ronidazole merits further investigation as a potential treatment for CDIs.

Association of fecal sample collection technique and treatment history with Tritrichomonas foetus polymerase chain reaction test results in 1717 cats.

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.

Efficacy and safety of ronidazole treatment against Tritrichomonas foetus in a cat colony with multiple disorders.

In a group of pedigree cats (n = 17) in poor health condition housed in an animal shelter in Vienna, Austria, with a history of persistent diarrhea, Tritrichomonas foetus infection was detected by PCR. Despite pre-existing clinical conditions all cats were treated with ronidazole (30 mg/kg PO q24h for 14 days) under close observation. After treatment, 11 of 14 initially positive animals remained negative for T. foetus during the observation period (six to eight weeks post treatment) and no diarrhea was observed. During treatment, nine cats showed mild to moderate neurological disorders (incoordination, mild tremor) at least once; six of these had already shown similar signs before treatment. Ronidazole treatment of multimorbid animals is acceptable if the benefit (here: clinical resolution and release from quarantine for adoption) is high. It is hypothesized that a high degree of inbreeding is a significant risk factor for the development of tritrichomonosis in cats.

The conundrum of feline Trichomonosis.

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.

Intestinal Tritrichomonas foetus infection in cats: a retrospective study of 104 cases.

The clinical presentation and response to treatment of cats infected with Tritrichomonas foetus have not been sufficiently described in a large number of pet cats. The aim of this study was to collect and analyze clinical data from pet cats diagnosed with intestinal T foetus infection. Clinical information was collected for 104 cats that tested polymerase chain reaction-positive for T foetus. The most common clinical sign was diarrhea (98%) with a median duration of 135 days (range 1-2880 days). Forty-nine of 83 (59%) cats had diarrhea since adoption. Other clinical signs included anorexia (22%), depression (24%), weight loss or failure to gain weight (20%), vomiting (19%), abdominal pain (9%) and increased appetite (3%). A total of 45 cats had completed treatment with ronidazole, 29 of which (64%) showed a good clinical response to treatment. Sixteen (36%) cats had either partial or no improvement, or a relapse shortly after discontinuation of treatment.

Control of Giardia infections with ronidazole and intensive hygiene management in a dog kennel.

Infections with the intestinal protozoan parasite Giardia in dogs and cats are common. Clinical signs vary from asymptomatic to small bowel diarrhea and associated discomfort. The control of infections in dogs is frequently a frustrating issue for animal owners and veterinarians. Drugs with antiprotozoal activity such as fenbendazole and metronidazole are recommended, however, they do not show 100% efficacy and superinfections occur regularly. Ronidazole is currently the drug of choice for the treatment of Tritrichomonas foetus in cats and there is now limited information available about its efficacy against Giardia spp. In the kennel investigated, dogs regularly showed loose feces and the presence of Giardia (assemblage C, renamed as G. canis) cysts. An elimination strategy of this parasite involving strict hygiene management and disinfection of the enclosures with 4-chlorine-M-cresol, oral treatment with ronidazole (30-50mg/kg BW bid for 7 days) and two shampooings (containing chlorhexidine) at the beginning and the end of the treatments was implemented for a group of 6 dogs. As a control another group of 7 dogs was transferred to the disinfected enclosures and shampooed, but left untreated. Dog feces were tested for the presence of Giardia cysts (SAF concentration technique) or Giardia antigen with a commercial ELISA (NOVITEC(®)) and a quick immunochromatography-based test (SensPERT(®)) before and between 5 and 40 days after the last treatment. All ronidazole-treated dogs were negative for Giardia cysts and antigen up to 26 days after the last treatment, while between 1 and 5 of the control animals tested positive in each of the test series. At this point, also dogs of the control group were again moved into clean enclosures, shampooed twice and treated with ronidazole. Five, 12 and 19 days after the last treatment, the dogs in the control group tested negative for Giardia cysts and antigen. However, all animals had again positive results at later time points in at least one of the three applied diagnostic techniques within 33-61 days after treatment. Furthermore, all dogs had episodes of diarrhea (for 1-4 days) within 14-31 days after treatment and unformed feces during the whole experiment. The positive effect of ronidazole against Giardia infections in dogs could be confirmed in this study. In particular, the combination of ronidazole treatment combined with the disinfection of the environment and shampooing of the dogs was highly effective in reducing Giardia cyst excretion and may therefore constitute an alternative control strategy for canine giardiosis.

Genetically different clonal isolates of Trichomonas gallinae, obtained from the same bird, can vary in their drug susceptibility, an in vitro evidence.

Trichomonas gallinae is a flagellated protozoon which parasitizes in the upper digestive tract of different birds, especially columbiformes (doves and pigeons) and falconiformes. The parasite is also a common inhabitant of the crop of psittacine birds and is frequently detected in budgerigars. The lesions associated with T. gallinae infection of the upper digestive tract range from mild inflammation of the mucosa to large caseous lesions that block the lumen of the oesophagus. Nitroimidazoles are considered to be the drugs of choice for the treatment of trichomonosis. However, only a few studies report the existence of resistant strains of T. gallinae to these drugs. Thus, in the present investigation cloned cultures of T. gallinae obtained from budgerigars and pigeons were analysed for the first time for their in vitro susceptibilities against four 5´-nitroimidazole derivates, including metronidazole, dimetridazole, ronidazole and ornidazole. Significantly different minimal lethal concentrations (MLCs) were observed for them against all four drugs. The lowest MLCs revealed the Trichomonas isolates obtained from two budgerigars, ranging from 2.0 ± 0.3 to 3.0 ± 0.7 µg/ml for metronidazole and dimetridazole, and from 2.0 ± 0.6 to 6.7 ± 1.7 µg/ml for ornidazole and ronidazole. Contrary to this, the highest MLCs were recorded for one Trichomonas isolate obtained from a pigeon, ranging from 83.3 ± 6.7 (for dimetridazole and ronidazole) to 103.3 ± 3.3 µg/ml (for metronidazole and ornidazole). The data obtained for the resistance testing were further compared with already available genetic data of the small subunit rRNA gene sequences and ITS-1, 5.8S rRNA and ITS-2 sequences, indicating a certain correlation between in vitro results and strain relationships.

Assessment of reproductive tract disease in cats at risk for Tritrichomonas foetus infection.

OBJECTIVE: To determine whether Tritrichomonas foetus infection resides in reproductive tract tissues from cats housed for breeding and for which a high prevalence of colonic T foetus infection has been reported. ANIMALS: 61 purebred cats in 36 catteries undergoing elective ovariohysterectomy or castration and for which reproductive tract tissues, feces, and a reproductive history were obtained. PROCEDURES: Reproductive tract tissues were examined for T foetus via light microscopy, immunohistochemical analysis, and PCR assay. History of reproductive tract disease was examined to detect statistical associations with identified or reported exposure to colonic T foetus infection. RESULTS: 15 of 61 (25%) cats and 22 of 33 (67%) catteries were identified with active or reported T foetus infection. Light microscopic, immunohistochemical, or molecular evidence of T foetus infection of the reproductive tract was not detected in any cats, including 15 cats with colonic T foetus infection, 29 cats residing in a cattery in which T foetus-infected cats were identified, and 8 cats for which gross or light microscopic evidence of reproductive tract disease was identified. There were no differences in total number of litters, number of litters per breeding, kitten mortality rate, or birth defects between cats or catteries infected with T foetus and those for which T foetus infection was not identified. CONCLUSIONS AND CLINICAL RELEVANCE: No evidence of reproductive tract colonization by T foetus was detected in this study. Accordingly, it is unlikely that reproductive tract infection with T foetus plays an important role in overall disease transmission.

[Tritrichomonas fetus: a new intestinal parasite in Swiss cats]. / Tritrichomonas fetus: Ein neuer Parasit im Darm von Schweizer Katzen.

Recent reports identified Tritrichomonas fetus, the causative agent of bovine trichomonosis, in cats with large-bowel diarrhea in the US. Between July 2007 and August 2008, a total of 105 Swiss cats were tested for T. fetus with the InPouchTM culture system and/or PCR, whereof 27 (26%) yielded positive results. All positive cats were pedigree cats, whereof 22 (81%) were less than 1 year of age (median 5 months). 25 (93%) of these cats lived in multi-cat households, and all but one were kept indoor. The clinical picture was dominated by large bowel diarrhea with increased frequency of defecation and fresh blood and mucus. Furthermore, inflamed anus and fecal incontinence was common. 52% of the T. fetus-positive cats were tested positive for Giardia before, but the treatment with fenbendazole or metronidazole only temporarily alleviated the clinical signs. The treatment with 30 mg/kg of ronidazole q12h p.o. was successful in all but 1 cat with only minor transient adverse effects in 3 cats. In conclusion, T. fetus has to be considered an important causative agent of large bowel diarrhea in cats in Switzerland, especially in young indoor pedigree cats.

Chronic intermittent diarrhea in a 14-month-old Abyssinian cat.

A 14-month-old intact, female Abyssinian cat was presented for chronic intermittent diarrhea and bilateral enlargement of the mammary glands. Gastrointestinal coccidiosis was diagnosed; therapy with sulfadi-methoxine was unsuccessful in the elimination of Isospora felis and clinical signs. Infection with Tritrichomonas foetus was diagnosed by fecal polymerase chain reaction (PCR) and successfully treated with ronidazole and dietary modification.

Tritrichomonas foetus infection in cats with diarrhoea in a rescue colony in Italy.

Faecal samples from 74 cats with chronic large bowel diarrhoea that were living in a rescue colony in Italy, were submitted for assessment of Tritrichomonas foetus by direct microscopy of faecal smears (n=20) and T foetus-specific culture (n=74), with confirmation by polymerase chain reaction (n=1). Twenty-four of the cats (32%) were found to be infected with T foetus. The infected cats were predominantly over a year of age (67%) and were all neutered non-pedigree domestic cats.

Efficacy of ronidazole for treatment of feline Tritrichomonas foetus infection.

OBJECTIVES: To determine the efficacy of ronidazole (RDZ), tinidazole (TDZ), and metronidazole (MDZ) against Tritrichomonas foetus in vitro and of RDZ for treatment of feline naturally occurring or experimentally induced T. foetus infection. ANIMALS: A cat naturally infected with T. foetus infection and diarrhea. Ten specific-pathogen-free (SPF) kittens. PROCEDURE: RDZ, TDZ, and MDZ were tested for activity against 3 different feline isolates of T. foetus in vitro. RDZ then was administered to a naturally infected cat at 10 mg/kg PO q24h for 10 days. SPF kittens were infected orogastrically with feline T. foetus and treated with either placebo or RDZ (10 mg/kg PO q12h for 14 days). Cats with relapsing infection or those receiving placebo were treated subsequently with RDZ (either 30 or 50 mg/kg PO q12h for 14 days). Feces were examined for T. foetus by direct microscopy, culture, and polymerase chain reaction (PCR) testing weekly. RESULTS: Both RDZ and TDZ killed T. foetus at concentrations >0.1 microg/mL in vitro. In the naturally infected cat, RDZ abolished diarrhea and T. foetus infection for 85 days after treatment, at which time infection and diarrhea relapsed. Retreatment with RDZ eradicated diarrhea and T. foetus infection for over 407 days. In experimentally induced infection, RDZ at 10 mg/kg caused initial improvement, but infection relapsed in all 5 cats 2 to 20 weeks after treatment. At 30 or 50 mg/kg, 10/10 cats were negative for T. foetus infection for follow-up durations of 21 to 30 weeks after treatment. CONCLUSIONS AND CLINICAL RELEVANCE: Oral administration of RDZ at 30 to 50 mg/kg q12h for 14 days resolved diarrhea and eradicated infection (on the basis of polymerase chain reaction [PCR] testing) in 1 naturally infected cat and 10 experimentally inoculated cats receiving a different isolate of T. foetus.

[The compatibility of ronidazol in racing pigeons in a controlled clinical study]. / Verträglichkeit von Ronidazol bei Brieftauben im kontrollierten klinischen Test.

Ronidazol is often used in racing pigeons for the treatment of Trichomonas infections and diseases. Therefore, in this study, the compatibility of the drug was examined by oral application over 7 days. For this purpose a randomized blind study was performed using four different groups (control group, 10 mg = therapy-group, 20 mg = double-dose-group and 40 mg = high-dose-group) of pigeons (Columba livia f. domestica) with 6 male and 6 female birds each. All birds were clinically healthy and between 6 and 12 weeks of age. The application of ronidazol at a dose of 10 mg/racing pigeon did show no side-effect within the duration of the study, e.g. no influence could be seen on clinical, haematological, blood-chemical and pathological parameters. Low-to middle-grade clinical alterations of the gastro-intestinal tract occurred in the high-dose group at day 6 and 7 of the application of the drug. Therefore a fourfold overdosing of ronidazol should be avoided.

Typhlitis caused by intestinal Serpulina-like bacteria in domestic guinea pigs (Cavia porcellus).

Between January 1992 and December 1996, Serpulina-like bacteria were demonstrated in intestinal tract lesions from 37 of 88 guinea pigs submitted to the University of Ghent in Ghent, Belgium, for necropsy because of disease and death from different unknown causes. All infected animals had a history of sudden death with minimal introductory clinical signs. Occasionally, they produced yellow, slimy feces or showed nervous signs, but the condition always had a fatal outcome within 24 h. When larger colonies of guinea pigs were involved, the disease spread very rapidly unless treatment with ronidazole was initiated. Lesions consisted of a catarrhal or hemorrhagic inflammation of the colon and cecum (typhlitis). Electron microscopy demonstrated the presence of large numbers of Serpulina-like organisms adhering to the cecal mucosae of these animals. Attempts to isolate the agents failed. The organisms did not stain by an immunofluorescence technique for the detection of Serpulina hyodysenteriae. The present data provide evidence that intestinal Serpulina-like organisms can be important as a cause of disease in guinea pigs.

Trichomonas gallinae in budgerigars and columbid birds in Perth, Western Australia.

OBJECTIVE: To estimate the prevalence of infection with Trichomonas gallinae and other parasites of the alimentary tract in psittacine and columbid birds in Perth and to determine in vitro the effectiveness of drugs commonly recommended for treating trichomoniasis. DESIGN AND PROCEDURES: Samples of crop contents were collected from aviary flocks of budgerigars (Melopsittacus undulatus) and other psittacine and columbid birds in both private and commercial collections in Perth. Similar samples from wild Senegal doves (Streptopelia senegalensis) also were collected. Crop contents were examined and cultured for Trichomonas gallinae and in vitro studies were conducted on the susceptibility of isolates to several drugs used commonly. Other parasites also were detected by faecal examination and/or necropsy. RESULTS: T gallinae was recovered from birds in 1 of 13 private collections of budgerigars (2/289 birds in total). Direct wet-mount examination of crop fluid identified 36.4% of samples at four commercial bird dealers which were later determined by culture to contain T gallinae. The prevalence of T gallinae infection range from 0 to 11.4% in budgerigars. The prevalence of T gallinae infection of wild Senegal doves was 46% and from one flock of racing pigeons was 59%. The in vitro minimum lethal concentrations of metronidazole, dimetridazole and ronidazole ranged from 40 to 96, 30 to 80 and 40 to 92 micrograms/mL respectively for six isolates of T gallinae. Other alimentary parasites detected during the survey included Spironucleus sp (syn. Hexamita sp), coccidia, Ascaridia platycerci and Raillietina sp. CONCLUSIONS: Thirteen budgerigar flocks belonging to members of avicultural societies in Perth had a low prevalence of trichomoniasis and other parasitic infections. The dose rate currently recommended for ronidazole may not result in complete protozoacidal activity against T gallinae infection.

Cochlosoma infections in finches.

OBJECTIVE: To survey finches in pet shops for Cochlosoma infection and evaluate the efficacy of antiprotozoal therapy with metronidazole or ronidazole. DESIGN: A survey of pet shop finches and drug efficacy trials. PROCEDURE: Finches in pet shops were randomly selected and their faeces examined microscopically for motile Cochlosoma sp trophozoites. Drug trials were carried out on 60 adult finches with naturally occurring infections. Body weight was measured and the faeces of each bird was examined for trophozoites at the beginning and 7 days after the end of treatment. In some birds, additional daily faecal examinations were done until three consecutive negative results were obtained. Metronidazole was administered at various dose rates by crop gavage or in drinking water to eight groups of five to ten finches each. Ronidazole was given in water for 7 days to ten finches. In addition, six finches whose faeces tested positive were necropsied and their tissues collected for histological examination. RESULTS: Motile flagellates in the faeces were identified as C anatis-like protozoa. Red-headed parrot-finches, Bengalese and Lady Gould finches were found to be most commonly infected. Cochlosoma sp was also found in the blue-faced parrot-finch, zebra finch, painted finch, nutmeg mannikin and double-barred finch. Metronidazole and ronidazole were found to be effective against Cochlosoma sp. Histological findings on infected adult finches were normal, except for the presence of numerous flagellates between the colorectal villi and cloacal mucosal folds. CONCLUSIONS: Cochlosoma anatis-like organisms can infect several species of finches and in adult finches are confined to the colorectum and cloaca. Infection in adult finches was mostly subclinical and could be treated effectively with metronidazole or ronidazole.

In vitro nitroimidazole resistance of Trichomonas gallinae and successful therapy with an increased dosage of ronidazole in racing pigeons (Columba livia domestica).

Six out of eight different Trichomonas gallinae strains isolated from racing pigeons proved to be resistant to the nitroimidazole drugs ronidazole, carnidazole and metronidazole. The minimal cytocidal concentration of ronidazole was determined in in vitro experiments. Moreover, a therapeutic dose for ronidazole was determined for the control of trichomoniasis in pigeons from which the resistant T. gallinae strains were isolated. It was a 5-fold increase of the recommended ronidazole dosage which eliminated the infection in affected pigeons.