Documentation of in vivo and in vitro aerobic resistance of feline Tritrichomonas foetus isolates to ronidazole.

BACKGROUND: The mainstays of treatment for clinically important trichomonad infections are the 5-nitroimidazoles. Metronidazole resistance of feline Tritrichomonas foetus is presumed because of common treatment failure, and tinidazole does not consistently eradicate infection. To date, ronidazole is the only drug demonstrated as effective for treatment of cats infected with T. foetus. OBJECTIVE: To document in vivo treatment failure and identify underlying causes and in vitro conditions of resistance of feline T. foetus to ronidazole. ANIMALS: Two intact male Abyssinians failing>or=5 courses of treatment with increasing doses of 5-nitroimidazole drugs. An intact male Abyssinian documented to clear infection after treatment with a single course of ronidazole. METHODS: T. foetus isolates were cultured from feces and tested in vitro for susceptibility to ronidazole under aerobic and anaerobic culture conditions. A urogenital nidus of T. foetus infection was investigated by culture, polymerase chain reaction, or immunohistochemical testing of urogenital specimens. RESULTS: Resistance to ronidazole under aerobic conditions was uniquely identified in T. foetus isolated from cats with well-documented treatment failure. Treatment failure could not be attributed to reinfection, inappropriate treatment protocol, or presence of a urogenital nidus of infection. CONCLUSIONS AND CLINICAL IMPORTANCE: Clinical resistance to metronidazole, low efficacy of tinidazole, and present documentation of in vivo and in vitro resistance to ronidazole in some cats are consistent with a high level of cross resistance of feline T. foetus to 5-nitroimidazole drugs. Current lack of alternative drugs with clinical efficacy against feline T. foetus suggests that active investigation of other treatment approaches is warranted.

Determination of the in vitro susceptibility of feline tritrichomonas foetus to 5 antimicrobial agents.

BACKGROUND: The nitroimidazole, ronidazole, has been demonstrated to have in vitro and in vivo activity against the protozoan Tritrichomonas foetus in cats. The purpose of this study was to evaluate the in vitro susceptibility of feline T. foetus isolates obtained from naturally infected cats to 5 antimicrobial agents and to compare the in vitro time kill of ronidazole and metronidazole. HYPOTHESIS: We hypothesized that nitroimidazoles have in vitro activity against T. foetus, whereas furazolidone, omeprazole, and paromomycin do not. ANIMALS: Fecal specimens were cultured from 4 naturally infected Bengal cats with a history of T. foetus-associated diarrhea. METHODS: A 24-hour susceptibility assay was performed on all 4 isolates for the 5 antimicrobial agents. A time-kill microdilution method was performed on 2 isolates for metronidazole and ronidazole. RESULTS: Paromomycin and omeprazole showed no in vitro effect at concentrations < or = 80 microg/mL. There was no significant difference in 24-hour susceptibilities among metronidazole, ronidazole, and furazolidone. In addition, only the results of the highest concentration tested (80 microg/mL) and concentrations of 1.25 and 2.5 microg/mL revealed significant differences in the rate of trophozoite killing, with ronidazole having a faster reduction in trophozoite survival. CONCLUSIONS AND CLINICAL IMPORTANCE: Time-kill assays demonstrated ronidazole had a higher lethal activity compared with metronidazole. These findings contrast with a previously published report and may reflect strain variation, different methodologies, or both. The lack of clinical response seen with metronidazole administration to treat feline trichomoniasis may not reflect inherent resistance but rather in vivo events involving drug distribution and pharmacokinetics.

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.

Susceptibility of Encephalitozoon cuniculi to several drugs in vitro.

In the light of the increased incidence of human Encephalitozoon infections and the absence of an established treatment protocol, a simple in vitro testing method to compare activities of drugs against Encephalitozoon cuniculi was developed. With this in vitro method, the 50% inhibitory concentrations of fumagillin, thiabendazole, albendazole, oxibendazole, and propamidine isethionate for E. cuniculi in rabbit kidney cells were determined. Itraconazole, toltrazuril, metronidazole, ronidazole, and ganciclovir were ineffective in this testing system.

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.

Minimal inhibitory concentrations of five antimicrobials against Treponema hyodysenteriae and Treponema innocens.

The minimal inhibitory concentrations of carbadox, dimetridazole, lincomycin, ronidazole, and tiamulin against isolates of Treponema hyodysenteriae and Treponema innocens were determined by an agar-dilution method. The results obtained indicated that tiamulin was the most effective antimicrobial in vitro against T. hyodysenteriae, followed by carbadox. Dimetridazole, lincomycin, and ronidazole had poor efficacy in vitro against the T. hyodysenteriae isolates. Isolates of T. innocens were more sensitive to the various antimicrobials. Carbadox and tiamulin were the most effective in vitro, followed by ronidazole, dimetridazole, and lincomycin.

Studies on the mechanism of activation and the mutagenicity of ronidazole, a 5-nitroimidazole.

Substantial evidence implicates the obligatory nucleophilic attack by water at C4 for the elimination of the carbamate and subsequent immobilization by electrophilic attack on protein thiols. Consequently, the strong correlation between the structural requirements for protein alkylation and for mutagenicity in TA100 suggests a possible role of nucleophilic addition at C4 or at the 2-methylene carbon for the expression of mutagenicity. Further studies directed at evaluating this possibility are currently in progress.

Drug residue formation from ronidazole, a 5-nitroimidazole. VI. Lack of mutagenic activity of reduced metabolites and derivatives of ronidazole.

The potential toxicity of ronidazole residues present in the tissues of food-producing animals was assessed using the Ames mutagenicity test. Since ronidazole is activated by reduction, reduced derivatives of ronidazole and metabolites formed by enzymatic reduction of ronidazole were tested for mutagenicity. When tested at levels several orders of magnitude higher than that at which ronidazole was mutagenic, 5-amino-4-S-cysteinyl-1,2- dimethylimidazole , a product of the dithionite reduction of ronidazole in the presence of cysteine, the 5-N-acetylamino derivative of ronidazole and 5-amino-1,2- dimethylimidazole all lacked mutagenic activity in Ames strain TA100. The metabolites of ronidazole formed by the incubation of ronidazole with microsomes under anaerobic conditions were also not mutagenic. These data demonstrate that although ronidazole is a potent mutagen, residues from it which may be present in the tissues of food-producing animals lack any mutagenic activity.

[Effect of ronidazole on the growth of weaned piglets and the levels of urea nitrogen in plasma and blood glucose]. / Wplyw ronidazolu na wzrost odsadzonych prosiat oraz poziom N-mocznikowego osocza i glukozy we krwi.

Four experiments were carried out on 412 piglets, reared in different environmental conditions in the period from 4 to 12 weeks of life. Ronidazole appeared to be the most valuable feed supplement among those studied: OTC, virginiamycin, LBC, SRE and bycitracin. In comparison with OTC it significantly improved weight increment of piglets from 8.1% to 15.4%, as well as utilization of feed from 4.0% to 9.1% with a significant decrease of morbidity and falls. The effect of the action of ronidazole was marked the strongest in young piglets directly after weaning, in less developed ones and those kept in unfavourable environmental conditions. Ronidazole did not cause changes in glucose concentration in blood, but in decreased the level of urea nitrogen in plasma. Mean concentration of urea nitrogen in piglets receiving ronidazole ranged from 16.9 to 17.6 mg/100 cm3, while in the control with OTC it ranged from 17.8 to 18.9 mg/100 cm3. A lower urea concentration in piglets in the group with ronidazole may indicate that, in comparison with OTC, it inhibited to a larger extent bacterial degradation of amino-acids and ureolysis of urea in tissues and alimentary canal content of piglets.

[The effect of Ridzol P used in pig fattening on carcass characteristics and meat quality]. / Vliv ridzolu P ve výkrmu prasat na nekteré charakteristiky tela zvírete a na jakost masa.

The meat and fat of slaughter pigs given diet with Ridzol P (60 mg per kg of feed) were studied. During the veterinary examination the additive was not found to have any adverse effect on the health of the animals. Judging from the studied meat and fat characteristics, there were no significant differences in the nutritive and technological value of the meat of the pigs given feed with Ridzol P, as compared with the controls. The additive was found to have a good influence on significantly higher weight of the cut lean parts. Hardly any difference was found in the proportions of the fatty acids of intramuscular fat, back fat and flare fat.

[Stimulatory and metabolic effects of Ridizol P in the fattening of pigs]. / Stimulacní a metabolické ucinky Ridzolu P u prasat ve vykrmu.

Fattened pigs (from 20 to 100 kg of body weight) given Ridzol P (6 g per 100 kg of mixture) were found to grow more quickly (by 8.1%) and to convert the feed mixtures more efficiently (A 1 by 6.6%, SOL by 3.9%). Twenty indicators of the metabolism were determined in serum, plasma, liver, and supra-renal gland after 110 days of Ridzol P administration. The changes in the concentrations of the total protein and urea suggest that the stimulative action of Ridzol P is based on better conversion of feed protein. The levels of vitamin A and E were increased in the blood plasma of the tested group. The content of vitamin A in the liver was significantly higher. The dry weight of the muscular and liver tissue of the tested animals was lower; the content of nitrogen compounds increased and the content of fat decreased in the muscular tissue of these animals.

Mutagenic evaluation of ronidazole.

Ronidazole was evaluated for mutagenic potential using in vitro microbial tests and in vivo studies in mice. The microbial test used the histidine requiring mutants of Salmonella typhimurium with and without a rat liver microsomal activation system (Ames test). The studies in mice included the dominant lethal test, micronucleus test and cytogenetic assays. Ronidazole was given orally in doses of 50, 100 and 200 mg/kg/day in the in vivo studies. In the dominant lethal test, groups of male mice were treated for five consecutive days before being mated with untreated females. In the micronucleus test, the mice were administered the compound for 2 or 5 consecutive days; they were killed 6 h after the last dose and bone marrow was examined for the presence of micronuclei in developing erythrocytes. In the cytogenetic assays, bone marrow cells in metaphase were examined for chromosome aberrations, 6, 24 and 48 h after mice were treated acutely with the test compound. In addition, similar examinations of chromosomes were made on mice given five consecutive dosages of ronidazole and killed 6 h after the last dose. The results of the various in vivo studies did not suggest that ronidazole would be mutagenic for the mammal. Ronidazole at concentrations of 10 and 50 mug/plate was found to increase the number of back mutations of missense mutants in the in vitro bacterial test. This finding confirms the results of Voogd et al. [19]. Incorporation of the microsomal activation system had no effect on the mutagenic capability of the test compound. In conclusion, although ronidazole was shown to be mutagenic in in vitro bacterial systems, the in vivo systems did not suggest that the compound would be mutagenic for the mammal.