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PubMed1986

Effect of twenty-three chemotherapeutic agents on the adherence and growth of Giardia lamblia in vitro.

The sensitivity of Giardia lamblia to 23 chemotherapeutic agents was evaluated in vitro with a Growth Inhibition Assay and a recently developed Adherence Inhibition Assay. Of the four established anti-giardia drugs, tinidazole, metronidazole, and furazolidone were found to have strong inhibitory effects on both growth and adherence, while mepacrine had a strong effect on growth only. Three drugs (mefloquine, doxycycline and rifampin) not previously used in giardiasis were found to have significant activity in vitro and may deserve consideration for clinical evaluation of efficacy. Also, the concurrent use of these two in vitro methods provided significant insights into the modes of action of some of these drugs on G. lamblia.

Animals↗
PubMedDate not supplied

Determination of anti-giardial activity in vitro by means of soluble formazan production.

A new microplate assay for the determination of activity in vitro against Giardia intestinalis has been developed in which viability is measured by soluble formazan production from a tetrazolium reagent. Metabolic reduction of the latter gives rise to a coloured product which may be determined directly by optical density measurement. Using this method the anti-giardial activities of more than 20 anti-microbial compounds have been assessed. The results obtained with a number of known anti-giardial agents, including metronidazole (50% inhibitory concentration [IC50] = 2.98 microM) and furazolidone (IC50 = 4.14 microM), compare well with previously published data; mepacrine, however, was found to produce a biphasic dose-response curve from which two IC50 values (0.816 and 6.83 microM) were obtained. This method provides a convenient means by which the search for new anti-giardial agents may be facilitated.

Animals↗
PubMed1993

Diarrhoeal disease: current concepts and future challenges. Antimicrobial therapy and prophylaxis.

Empirical antimicrobial therapy is indicated in patients with diarrhoea who have high fever and systemic toxicity, dysenteric disease, or travellers' diarrhoea. Antimicrobials are essential for those with severe shigellosis and amoebiasis. They are useful or possibly useful for other forms of diarrhoeal disease including amoebiasis (milder forms), campylobacteriosis, cholera, giardiasis, shigellosis, and diarrhoea due to a variety of other laboratory-defined bacterial enteropathogens. Furazolidone is useful in infantile giardiasis and mildly effective in other forms of bacterial diarrhoea. Trimethoprim/sulphamethoxazole is effective against Shigella spp. in ost parts of the world. Erythromycin is considered the treatment of choice for campylobacteriosis. For adults, the quinolone antimicrobials represent the most useful class of drugs for bacterial enteropathogens. Several dilemmas currently exist in the area. They include the lack of drugs for the therapy of trimethoprim-resistant shigellosis in children, overuse of antimicrobials in the developing world, and the potential for post-treatment prolongation of intestinal excretion of non-typhoid salmonellae. Antimicrobial chemoprophylaxis can be used in the rare person from an industrialized area during brief travels to a tropical region who has a serious underlying medical problem, cannot exercise care in what is eaten and drunk, and will have the purpose of the trip put at jeopardy should any illness develop (even that rendered short-term by effective therapy). For most people, therapy of illness is preferred to prophylaxis.

Adult↗
PubMed1987

Treatment of cryptosporidiosis with oral bovine transfer factor.

Cryptosporidia are intestinal protozoans long known to cause diarrhea in humans, especially those with acquired immune deficiency syndrome (AIDS). When transfer factor prepared from calves which possessed delayed-type hypersensitivity to Eimeria bovis was given to nonimmune calves and mice it conferred protection against clinical infection (coccidiosis). Recent studies with oral bovine transfer factor have shown that it can confer cell-mediated immunity to humans. Based on these findings we decided to treat eight AIDS patients suffering from Cryptosporidium-associated diarrhea with transfer factor prepared from calves immune to Cryptosporidium. Prior to treatment with transfer factor, three patients had been treated with spiramycin, one patient with alpha-difluoromethylornithine (DFMO), and one patient with furazolidone for greater than 1 month without clinical or laboratory improvement. Following administration of transfer factor, five or eight patients exhibited a decrease in the number of bowel movements and the development of formed stools. Cryptosporidium was eradicated from the stools of four patients but two of these patients subsequently relapsed and one patient continued to have diarrhea despite the absence of Cryptosporidium in the stool. One patient has been free of diarrhea and Cryptosporidium for 2 years after discontinuation of transfer factor therapy.

Acquired Immunodeficiency Syndrome↗
PubMed1986

Comparison of methods for routine separation of coagulase-negative staphylococci from micrococci isolated from sheep.

A total of 176 Gram-positive, catalase positive cocci strains, isolated from sheep were studied by different routine tests for the differentiation of staphylococci and micrococci, comparing their validity and usefulness. By glucose fermentation and growth in the anaerobic portion of thioglycolate 85 and 73.6% respectively of coagulase negative staphylococci were misclassified as Micrococcus spp. Susceptibility to lysostaphin was an adequate test for the differentiation of the strains. Atypical results in the production of acid from glycerol/erythromycin were obtained in 11.8% of the coagulase negative strains and 16.7% of micrococci. The combined use of the selective media furazolidone agar and Schleifer and Krämer medium resulted in a fast and useful separation of ovine staphylococci and micrococci. The bacteriolytic activity misclassified 32.2% of the coagulase negative strains.

Animals↗
PubMed1990

Biotypes, serogroups and antibiotic susceptibility of Campylobacter jejuni and Campylobacter coli in Chile.

Phenotypic markers were studied in 105 strains of thermophilic campylobacters isolated from human beings, animals and drinking water in Santiago, Chile. The strains were identified as Campylobacter jejuni (n = 49) and Campylobacter coli (n = 56). Biotypes I and II (Lior schema) accounted for 96% C. jejuni isolates, the other 4% being biotype IV but the two biotypes of C. coli were about equally represented. A total of 28 serogroups (Lior's heat-labile antigens) were identified. Lior 13, 9, 79, 2 and 4 were prevalent among the C. jejuni, while Lior 8, 21 and 29/75 were prevalent among the C. coli isolates. These serogroups accounted for 73% all isolates. The distribution of biotypes and serogroups in patients and asymptomatic persons were similar. Human campylobacters were often resistant to ampicillin (31%) but sensitive to erythromycin and furazolidone. Swine C. coli isolates proved resistant to streptomycin (46%), tetracycline (38%) and erythromycin (15%). Determination of phenotypic and serological characters provides valuable epidemiological markers in the study of campylobacter infections.

Animals↗
PubMed1979

Action of nitrofurans on E. coli: mutation and induction and repair of daughter-strand gaps in DNA.

The antibacterial and mutagenic potency of 9 nitrofurans in "treat and plate" experiments varied over almost 5 orders of magnitude. The relative toxicities were as follows: FANFT greater than AF2 greater than ANFT greather than furazolidone greater than furagin greater than nitrofurantoin greater than nitrofurazone greater than methylnitrofuroate greater than nitrofuroic acid. In general, mutagenic activity paralleled toxicity. The compounds at concentrations corresponding to their LD50's, induced mutations at frequencies which ranged from 2.5/10(6) survivors for FANFT to 130/10(6) survivors for furagin (NF416). The observed differences in antibacterial and mutagenic activity are unlikely to be due to lack of activation of the weaker agents since the two most potent agents were reduced somewhat more slowly than many of the less active agents. The relative sensitivities to the antibacterial effects of AF2 of strains WP2, WP2 uvrA, CM561 (lexA) and CM571 (recA) were 1 : 1.6 : 3 : 7 and to nitrofurazone 1 : 1 : 25 : 50. The wvrA strain was 6--7-fold more mutable with both these agents than was WP2. No increase over the spontaneous mutation frequency was observed when recA or lexA strains were exposed to either AF2 or nitrofurazone in these experiments. When wild-type of wvrA bacteria containing nitrofuran-induced lesions replicated their DNA in drug-free medium in the presence of [3H]thymidine for 5 min, the label was found in low molecular weight DNA indicating that daughter-strand gaps were formed. During subsequent incubation in nonradioactive medium the molecular weight of the DNA increased to the control value. A recA strain (which was very sensitive to the lethal effects of AF2 and nitrofurazone) lacked the ability to repair daughter-strand gaps caused by nitrofuran-induced lesions.

DNA Repair↗
PubMed1992

On the induction of umu gene expression in Salmonella typhimurium strain TA1535/pSK1002 by some nitrofurans.

Several nitrofurans were found to induce umu gene expression in Salmonella typhimurium TA1535/pSK1002 as defined on the basis of at least a 2-fold increase of beta-galactosidase activity over the background level. beta-Galactosidase activity increased with increasing concentrations of the chemical, attained a maximum at a concentration which was different for different nitrofurans used, and then gradually decreased with a further increase of the nitrofuran concentration. The umu gene expression test revealed that the genotoxic activity was highest for furazolidone and lowest for 5-nitro-2-furaldehyde.

Azides↗
PubMed1987

Mutagenicity of nitrofurans in Salmonella typhimurium TA98, TA98NR and TA98/1,8-DNP6.

8 representative 2-substituted 5-nitrofurans were assayed for mutagenicity in Salmonella typhimurium strains TA98, TA98NR and TA98/1,8-DNP6. The tested compounds were: 5-nitro-2-furanacrylic N-(5-nitro-2-furfurylidene)hydrazide (1); furazolidone (2); 5-nitro-2-furanacrolein (3); 5-nitro-2-furaldehyde semicarbazone (4); 5-nitro-2-furaldehyde (5); nitrofurantoin (6); 5-nitro-2-furaldehyde diacetate (7); and 5-nitro-2-furoic acid (8). These compounds exhibited markedly different mutagenic activities in TA98, and these mutagenicities were similar both in the presence and the absence of rat-liver hepatic S9 activation enzymes. The mutagenic responses ranged from potent (90-300 revertants/nmole, compounds 1-3), to medium (about 10 revertants/nmole, compounds 4 and 6), to weak (0-4 revertants/nmole, compounds 5, 7 and 8). The mutagenicity of 3 was similar in all 3 tester strains, while compound 8 was essentially inactive. The mutagenicities of 1, 4, 5 and 7 were decreased 30-75% in TA98NR, while 2 and 6 showed an even greater depression of activity in this strain. Compound 6 with S9 was about equally mutagenic in TA98 and TA98/1,8-DNP6, while the activities of 6 without S9 and 2 and 7 both with and without S9 were 50-75% lower in TA98/1,8-DNP6. Compounds 1, 4 and 5 were only about 5-10% as mutagenic in TA98/1,8-DNP6 as in TA98. These results suggest that: (i) nitrofurans and their S9-mediated metabolites have similar mutagenic potencies; (ii) with the possible exception of No. 3, nitroreduction is the major route of mutagenic activation for these nitrofurans; and (iii) for compounds 2, 6 and 7, both the presumed N-hydroxy and N,O-ester derivatives of the corresponding aminofuran metabolites appear to lead to mutations.

Dose-Response Relationship, Drug↗
PubMed1995

Free radical detoxification in Giardia duodenalis.

Non-denaturing polyacrylamide gels were used to analyse superoxide dismutase (SOD), catalase, peroxidase, NADH oxidase and NADH peroxidase in the microaerophilic protozoan parasite Giardia duodenalis. A cytosolic H2O-producing NADH oxidase and membrane-associated NADH peroxidase were readily detected from G. duodenalis. In all Giardia strains investigated the NADH oxidase was present in high levels (1.2-2 U (mg protein)-1). Using the same technique, NADH oxidase activity was also detected in the microaerophilic protozoan parasites Tritrichomonas foetus, Trichomonas vaginalis and Entamoeba histolytica and in the bacterium Escherichia coli. The conventional enzymes of oxidative stress management (superoxide dismutase, catalase and peroxidase) were not detected in particulate or cytosolic extracts from recent and established strains of Giardia assayed in situ. Spectrophotometric assays also yielded negative results. The same methodology readily detected one or more of these enzyme activities in T. foetus, T. vaginalis and E. coli. Superoxide dismutase activity was not detected in lines of Giardia resistant to high levels of metronidazole or furazolidone. Furthermore, the agents 1,10 phenanthroline, diamide, MnCl2 and KNO3, which induce SOD in anaerobically cultured E. coli, did not induce SOD in Giardia. 1,10 phenanthroline has also been shown to induce iron-containing (Fe-) SOD in Entamoeba. Neither peroxidase nor catalase activities were detected in a peroxide-resistant line of Giardia. Viable trophozoites from parent lines were able to decompose H2O2 at a significant rate. It appears that the conventional SOD, catalase and peroxidase utilised in aerobic metabolism have been substituted in Giardia by NADH oxidase and NADH peroxidase, similar to anaerobic bacteria. The O2-scavenging NADH oxidase explains the previously observed futile 'respiration' in Giardia.

Animals↗
PubMed1996

Characterisation and purification of pyruvate:ferredoxin oxidoreductase from Giardia duodenalis.

The major 2-oxoacid oxidoreductase (2-OR), pyruvate:ferredoxin oxidoreductase (PFOR) from Giardia duodenalis has been purified to apparent homogeneity. A second 2-OR with a preference for alpha-ketobutyrate as substrate was identified and was removed from PFOR containing fractions during purification. Only PFOR and the second 2-OR were identified in gels of crude Giardia extracts assayed for 2-OR activity. The native form of PFOR which is membrane associated, is a homodimer of 138 kDa subunits. Pyruvate is the preferred substrate: alpha-ketobutyrate and oxaloacetate, but not phenyl-pyruvate or alpha-ketoglutarate, are decarboxylated. PFOR from Giardia is more stable than PFOR from most other organisms and purified PFOR can be stored without deterioration at -70 degrees C. Purified PFOR donates electrons to Giardia ferredoxin (Fd I) with concomitant reduction of metronidazole. However, two other Giardia ferredoxins did not accept electrons from PFOR. Consistent with the involvement of PFOR in metronidazole activation, the activity of pyruvate dependent 2-OR activity was decreased in all metronidazole-resistant lines tested but not in furazolidone-resistant lines. The presence of three different ferredoxins and two 2-ORs in Giardia suggests that a number of different electron transport pathways operate in this organism providing unusual metabolic flexibility for a eukaryote.

Animals↗
PubMed1995

In vitro susceptibility to antimicrobial drugs of 62 Salmonella strains isolated from horses in The Netherlands.

The in vitro activity of 17 antimicrobial drugs against strains of Salmonella typhimurium (n = 52), Salmonella thompson (n = 2), Salmonella heidelberg (n = 3), Salmonella hadar (n = 2), Salmonella enteritidis (n = 1), Salmonella infantis (n = 1) and Salmonella derby (n = 1) was tested using the agar dilution method. The strains were isolated from horses admitted to the Large Animal Clinics of Utrecht University. The majority of strains were susceptible to gentamicin, amikacin, kanamycin, enrofloxacin, ciprofloxacin, flumequine, colistine, furazolidone and ceftiofur. However, all strains of Salmonella typhimurium phage type 200 (n = 14), were multiresistant i.e. were resistant to ampicillin amoxycillin, amoxycillin in combination with clavulanic acid, chloramphenicol, nitrofurantoin, trimethoprim, aditoprim and baquiloprim. Two of these strains were also resistant to gentamicin. Based on the susceptibility data found in the present study in combination with pharmacokinetic data available in the literature a rationale for antimicrobial therapy in equine salmonellosis is given. As first choice, gentamicin at a dosage of 3 mg/kg combined with ampicillin at a dosage of 20 mg/kg given with a 8-12 hour dosing interval by intravenous route is advised. As an alternative, the intravenous administration of trimethoprim/sulfonamide combinations given twice daily at a combined dose of 30 mg/kg is suggested.

Animals↗
PubMedDate not supplied

Stomatococcus mucilaginosus bacteremias. Typical case presentations, simplified diagnostic criteria, and a literature review.

Even though Stomatococcus mucilaginosus is considered indigenous oral-pharyngeal flora, cited literature and case reports indicate that it can be the cause of infectious conditions. Tested strains were isolated from blood, the oral region, and wound sources. The organism was routinely misidentified or not identified by conventional or commercial systems (Vitek, STAPH-Trac). Four antimicrobial diagnostic disks for example, bacitracin (0.04 units; Taxo A), furazolidone (100 micrograms), novobiocin (5 micrograms), and polymyxin B (300 units), were evaluated as possible addition to previously applied biochemical characteristics that differentiate between S. mucilaginosus, Micrococcus sp., and coagulase-negative staphylococci. Consistent antimicrobial susceptibility patterns among our isolates to the diagnostic disks produced applicable characteristics for discriminating S. mucilaginosus from similar microorganisms. However, therapeutic choices of antimicrobial agents should be guided by individual organism susceptibility test results because of variable, often resistant patterns to beta-lactams, aminoglycosides, macrolides, new fluoroquinolones, and sulfonamides.

Adult↗
PubMed2005

Quantitative determination of four nitrofuran metabolites in meat by isotope dilution liquid chromatography-electrospray ionisation-tandem mass spectrometry.

A confirmatory method based on isotope dilution liquid chromatography-tandem mass spectrometry (LC-MS/MS) has been developed for the low-level determination of residues of four nitrofuran veterinary drugs in meat, e.g., furazolidone, furaltadone, nitrofurantoin, and nitrofurazone. The procedure entails an acid-catalysed release of protein-bound metabolites, followed by their in situ conversion into the 2-nitrobenzaldehyde (NBA) imine-type derivatives. Liquid-liquid extraction and clean-up on a polymeric solid phase extraction cartridge are then performed before LC-MS/MS analysis by positive electrospray ionisation (ESI) applying multiple reaction monitoring of three transition reactions for each compound. Reliable quantitation is obtained by using one deuterated analogue per analyte (d4-NBA derivative) as internal standard (IS). Validation of the method in chicken meat was conducted following the European Union (EU) criteria for the analysis of veterinary drug residues in foods. The decision limits (CCalpha) were 0.11-0.21 microg/kg, and the detection capabilities (CCbeta) 0.19-0.36 microg/kg, thus below the minimum required performance limit (MRPL) set at 1 microg/kg by the EU. The method is robust and suitable for routine quality control operations, and more than 200 sample injections were performed without excessive pollution of the mass spectrometer or loss of LC column performance.

Chromatography, Liquid↗
PubMed2005

A variant type of Vibrio cholerae SXT element in a multidrug-resistant strain of Vibrio fluvialis.

Vibrio fluvialis strain H-08942 was isolated from an infant aged 6 months who was suffering from cholera-like diarrhea in India. This strain showed the typical multidrug-resistance phenotype of an SXT element. It was resistant to sulfamethoxazole (Su), trimethoprim (Tm), chloramphenicol (Cm) and streptomycin (Sm), in addition to other antibiotics such as ampicillin (Am), furazolidone (Fz), nalidixic acid (Na), and gentamicin (Gm). The SXT element is a Vibrio cholerae-derived integrative and conjugative element (ICE) that has also been referred to as a conjugative transposon. Our goal was to find a relationship between these resistant phenotypes and the presence of the SXT element in this unique strain. By using PCR, we detected the antibiotic resistance genes, the integrase gene and the attP attachment site of SXT element. Cloning and DNA sequencing results showed that both the SXT integrase gene and its attP site of V. fluvialis were similar but not identical to those of V. cholerae. The SXT integrase gene of V. fluvialis has a 99% identity to that of V. cholerae, and the attP site of SXT of V. fluvialis is variant and shorter (641 bp) than that of V. cholerae (785 bp). It was possible for the SXT of V. fluvialis to be transferred by conjugation to a laboratory strain of Escherichia coli. Here, we report the detection of a variant SXT element in species other than V. cholerae, with molecular characterization and analysis of its integrase gene and its attP site.

Conjugation, Genetic↗
PubMed2004

Acute lymphoblastic leukemia presenting with urticarial plaques and hypereosinophilia in a child.

Our report describes a previously healthy 10-year-old female who was seen for urticarial plaques and mild loss of appetite. An initial laboratory workup revealed an elevated leukocyte count of 30,000/microL and a peripheral eosinophil count of 22,500/microL. A skin biopsy showed a marked hypersensitivity tissue response with abundant eosinophils. Further investigation of her peripheral eosinophilia uncovered Giardia lamblia in a stool sample. Despite treatment with the anti-parasitic agent furazolidone, the patient's urticarial plaques, leukocyte count, and peripheral eosinophil count remained unchanged. A bone marrow biopsy confirmed a diagnosis of acute lymphoblastic leukemia (ALL). ALL with hypereosinophilia (ALL/Eo) represents a rare and distinct subset of ALL, with more than 30 cases documented in the literature. Our discussion summarizes the clinical aspects of this disease and reviews the reported dermatological manifestations of ALL/Eo.

Child↗
PubMed2005

Determination of nitrofuran metabolites in poultry muscle and eggs by liquid chromatography-tandem mass spectrometry.

The use of nitrofurans in food-producing animals has been banned in EU. Detection of the protein-bound nitrofuran metabolites is the best approach to evaluate their utilization. A fast, sensitive and reliable LC-MS-MS method is presented to analyze simultaneously the metabolites of four commonly used nitrofuran drugs, furazolidone, furaltadone, nitrofurazone and nitrofurantoin. The sample clean up was performed by a single liquid-liquid extraction step, after a hydrolysis and derivatisation process. Separation of the molecules was performed by liquid chromatography in a C18 column (100 mmx2.1 mm, 4 microm) at room temperature. The quantitative and confirmatory determination of these metabolites was performed by multiple reactions monitoring (MRM). Limits of quantification of 0.5 ngg(-1) were achieved and the total analysis was accomplished in 5 min. This protocol has been applied to identify contaminated samples of poultry muscle and egg products.

Animals↗
PubMed2006

Phenotypic and molecular typing of Vibrio cholerae O1 and O139 isolates from India.

OBJECTIVES: To determine whether Vibrio cholerae strains with similar phage types are also genetically homogeneous or heterogeneous by molecular typing methods like PFGE and RAPD-PCR employed in this study. METHODS: A total of 26 V. cholerae O1 and O139 strains received from different parts of India were analyzed by using conventional phage typing method, antibiogram and molecular typing methods such as PFGE and RAPD-PCR. RESULTS: Both O1 and O139 strains were resistant against two antibiotics (Ampicillin and Furazolidone) were detected. All of these strains were clustered in a single phage type, i.e., Type 27 for V. cholerae O1 and Type 1 for V. cholerae O139. Extensive molecular characterization by RAPD and PFGE showed that six sets of O1 and O139 strains, each comprising two strains, had identical PFGE and RAPD profiles. Only one O139 strain (PL-4) had unique RAPD and PFGE profile among all the 26 V. cholerae strains used in this study. CONCLUSION: Apart from serology, the strains of V. cholerae can be discriminated by this conventional phage typing system that offers the basic information on identification, biotyping and discrimination of strains. But, a high level of heterogeneity was observed in RAPD and PFGE profiles indicating the clonal diversity of V. cholerae O1 and O139 strains. It was concluded that these strains were phenotypically identical through phage typing system and antibiogram but genetically dissimilar, as shown in molecular typing systems.

Anti-Bacterial Agents↗