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Effects of danofloxacin or consecutive fluoroquinolone and macrolide treatments on resistance development in Campylobacter have remained uncharacterised. Therefore we analysed the development of resistance in porcine Campylobacter coli before and after danofloxacin and tylosin treatments at a farrowing farm. Danofloxacin-treated (n=12, group A) and control pigs (n=15, group B) were subsequently treated with tylosin and sampled longitudinally. C. coli were isolated and susceptibilities to ciprofloxacin and erythromycin were assessed, isolates were genotyped with PFGE and resistance-related mutations were identified. Isolates from the danofloxacin-treated pigs had more frequently non-wild type MICs (above the epidemiological cut-off value (ECOFF)) for ciprofloxacin (P<0.001) and erythromycin (P<0.05) than those isolated before danofloxacin or those from the controls. Subsequent tylosin treatment increased proportion of isolates with non-wild type MICs for erythromycin in both groups A and B (P<0.01) and, interestingly, proportion of isolates with non-wild type MICs for ciprofloxacin in group B (P<0.001) with high MICs (128 μg/ml). PFGE analysis revealed treatments selecting predominant genotypes with variable resistance patterns and decreasing initial diversity of genotypes. The most common genotype had mainly high MICs for ciprofloxacin among danofloxacin-treated pigs but wild type MICs (below the ECOFF) among the controls housed in the same pens. This suggests that the non-wild type isolate was rarely transmitted or outcompeting wild type genotype in the control pigs without selection pressure. Isolates exhibiting non-wild type MICs for ciprofloxacin harboured the C257T (Thr-86-Ile) mutation in the gyrA gene. In conclusion, a high dose of danofloxacin used at the farm did not prevent emergence of isolates with high MICs for ciprofloxacin. After subsequent tylosin treatment isolates had even higher MICs for ciprofloxacin and erythromycin than before the treatment. Therefore, controlled use of antimicrobials in food animal production is essential., (Copyright © 2011 Elsevier B.V. All rights reserved.)

Limited knowledge is available regarding the dynamics of macrolide resistance under farm conditions with natural Campylobacter populations. We examined the dynamics of antimicrobial resistance in Campylobacter coli at a large pig farm. Faeces were sampled from untreated sows and piglets (n=57), weaned pigs treated with tylosin (n=68) and pigs of the same group 3-5 weeks after withdrawal of tylosin (n=15). Additionally, 48 weaned pigs were sampled after tylosin had not been administered for 7 months at the farm. MICs for seven antimicrobials were determined, isolates were genotyped by PFGE and mutations conferring macrolide resistance were identified. Resistance to at least one antimicrobial agent was higher (P<0.001) in the isolates from the treated pigs (30 of 56) than in those from the untreated animals (2 of 40). Resistance to ciprofloxacin, erythromycin, nalidixic acid and streptomycin was higher (P<0.05) in the isolates from the treated pigs than in those from the untreated animals. All 14 erythromycin-resistant isolates studied (MIC ≥ 512 μg/ml) contained mutation A2075G in 23S rRNA. Resistance against at least one antimicrobial was significantly lower (P<0.05) when tylosin had not been administered for 7 months. Resistance to erythromycin and streptomycin also decreased (P<0.05). PFGE analysis revealed a change of genotypes induced by tylosin treatment. In conclusion, tylosin treatment of pigs selected for a high-level of resistance to erythromycin and resistance to ciprofloxacin, nalidixic acid and streptomycin also increased in C. coli isolates within a few days., (Copyright © 2010 Elsevier B.V. All rights reserved.)

Previously we have shown that experimentally infected swine, fed an antibiotic-free diet, can become colonized and shed Escherichia coli O157:H7 for at least 2 months. However, in epidemiological studies this organism is only rarely recovered from domestic swine and the basis for this discrepancy is not clear. In this report we demonstrate that significantly fewer pigs fed diets containing subtherapeutic levels of either tylosin or chlorotetracycline shed E. coli O157:H7 for longer than 2 weeks compared to those fed an antibiotic-free diet. In contrast to tylosin and chlorotetracycline, the addition of bacitracin methylene disalicylate to the diet did not influence the recovery of E. coli O157:H7. These results suggest that some antibiotics may alter the gastrointestinal tract flora in ways that create a less favorable environment for E. coli O157:H7 in swine., ((c) 2009 Elsevier B.V. All rights reserved.)

American Foulbrood (AFB) of honeybees (Apis mellifera L.), caused by the Gram-positive bacterium Paenibacillus larvae is one of the most serious diseases affecting the larval and pupal stages of honeybees (A. mellifera L.). The aim of the present work was to asses the response of 23 strains of P. larvae from diverse geographical origins to tilmicosin, a macrolide antibiotic developed for exclusive use in veterinary medicine, by means of the minimal inhibitory concentration (MIC) and the agar diffusion test (ADT). All the strains tested were highly susceptible to tilmicosin with MIC values ranging between 0.0625 and 0.5 microg ml(-1), and with MIC(50) and MIC(90) values of 0.250 microg ml(-1). The ADT tests results for 23 P. larvae strains tested showed that all were susceptible to tilmicosin with inhibition zones around 15 microg tilmicosin disks ranging between 21 and 50mm in diameter. Oral acute toxicity of tilmicosin was evaluated and the LD(50) values obtained demonstrated that it was virtually non-toxic for adult bees and also resulted non-toxic for larvae when compared with the normal brood mortality. Dosage of 1000 mg a.i. of tilmicosin applied in a 55 g candy resulted in a total suppression of AFB clinical signs in honeybee colonies 60 days after initial treatment. To our knowledge, this is the first report of the effectiveness of tilmicosin against P. larvae both in vitro and in vivo.

The objective of this study was to determine whether early detection and treatment with tilmicosin would reduce the number of yearling bulls culled due to vesicular gland disease at the time of their first semen test. Bulls (n=2207) of 15 breeds at 17 performance test stations were examined by transrectal palpation at 9-12 months of age and assigned to three treatment groups: (1) positive treated group, receiving subcutaneous injections of tilmicosin every second day for three treatments (2) positive untreated control group, (3) negative untreated control group. Transrectal palpation of the glands was done again at a pre-sale semen test 28-70 d (mean=42.8 d) after the first examination. Semen was evaluated for the presence of pus and/or leukocytes by light microscopy. The proportion of bulls with vesiculitis was 4.4% (97/2207). At the pre-sale semen test the number of bulls with vesiculitis had decreased to 1.3% (29/2207); however, seven of these were new cases that developed after the first examination. Therefore, there was a recovery rate of 75/97 (77.3%) in the original group of bulls positive for vesiculitis. At the pre-sale test, there was no difference in the proportion of bulls with enlarged glands in the positive treated group (15/66) versus the positive untreated group (7/31) and no difference in the proportion of bulls with leukocytes or pus in their semen samples. The results of this experiment did not indicate any advantage in detection and treatment of vesiculitis before a year on age due to a high rate of spontaneous recovery.

The minimal inhibitory concentrations (MICs) of tylosin were determined to 67 strains of Paenibacillus larvae subsp. larvae, the causal agent of American Foulbrood (AFB) disease, from different geographical origins. MIC values obtained ranged from 0.0078 to 0.5 microg/ml. These very low values imply that no resistance to tylosin was found in any isolate of the Foulbrood pathogen. The measurement of diseased larvae with AFB-clinical symptoms in three different field studies demonstrated that tylosin treatment could be effective in vivo. No negative effects in colonies were noted at any dosage rates or forms of application. These studies demonstrate that tylosin, as tartrate, can be used to treat AFB in honeybee colonies.

Clostridium difficile and Clostridium perfringens are anaerobic, Gram-positive bacilli that are common causes of enteritis and enterotoxemias in both domestic animals and humans. Both organisms have been associated with acute and chronic large and small bowel diarrhea, and acute hemorrhagic diarrheal syndrome in the dog. The objective of this study was to determine the in vitro antimicrobial susceptibilities of canine C. difficile and C. perfringens isolates in an effort to optimize antimicrobial therapy for dogs with clostridial-associated diarrhea. The minimum inhibitory concentrations (MIC) of antibiotics recommended for treating C. difficile (metronidazole, vancomycin) and C. perfringens-associated diarrhea in the dog (ampicillin, erythromycin, metronidazole, tetracycline, tylosin) were determined for 70 canine fecal C. difficile isolates and 131 C. perfringens isolates. All C. difficile isolates tested had an MIC of or=256 microg/ml for both erythromycin and tylosin. A third C. perfringens isolate had an MIC of 32 microg/ml for metronidazole. Based on the results of this study, ampicillin, erythromycin, metronidazole, and tylosin appear to be effective antibiotics for the treatment of C. perfringens-associated diarrhea, although resistant strains do exist. However, because there is limited information regarding breakpoints for veterinary anaerobes, and because intestinal concentrations are not known, in vitro results should be interpreted with caution.