Antimicrobial and heavy metal resistance in commensal enterococci isolated from pigs

Antibiotic resistance in animal isolates of enterococci is of public health concern because of the risk of transfer of antibiotic resistance isolates or resistance determinants to consumers via the food chain. In this study, phenotypic and genotypic resistance in 192 pig isolates of enterococci to ampicillin, avilamycin, avoparcin, bacitracin, flavophospholipol, gentamicin, narasin, tetracycline, tiamulin, tylosin, vancomycin, virginiamycin, copper and zinc were investigated by susceptibility test and molecular methods. Resistance rates varied between the species but all isolates were susceptible to ampicillin, avilamycin, avoparcin, gentamicin and narasin but resistant to tetracycline and tylosin and intermediately resistant to copper. Only Enterococcus gallinarum and Enterococcus casseliflavus were resistant to vancomycin and virginiamycin resistance was present in less than half the Enterococcus faecium isolates. Zinc resistance was largely confined to Enterococcus faecalis but bacitracin resistance was uncommon in E. faecalis in comparison with the other species. Tiamulin resistance was common in all species except E. casseliflavus. Resistance to flavophospholipol was detected in most E. faecium isolates and in a high proportion of E. gallinarum, E. casseliflavus and E. hirae/durans but was only found in one isolate of E. faecalis. No tetO, rplC, rplD, vanA, vanB, vatA and vatD genes were found. The presence of ermB, tetL, tetM, tcrB, aac6-aph2, tetK, tetS, vanC1, vanC2, lsaA, lsaB and vatE varied between the species and largely corresponded to the susceptibility phenotype. The findings show that resistance to antibiotics of high clinical significance for nosocomial Enterococcus infections is absent, whereas antimicrobial resistance was detected for some other antibiotics including bacitracin, flavophospholipol, tetracycline, tiamulin, tylosin and virginiamycin.     

In vitro lactic acid inhibition and alterations in volatile fatty acid production by antimicrobial feed additives

Batch culture fermentations were used to determine the effects of avoparcin, lasalocid, monensin, narasin, salinomycin, thiopeptin, tylosin, virginiamycin, monensin + tylosin combination, and two new ionophore compounds (RO22-6924/004 and RO21-6447/009) on lactic acid and volatile fatty acid (VFA) production. Ruminal fluid from cattle fed a high alfalfa hay diet was incubated with glucose for 12 h in a buffered medium to determine the effect of antimicrobial compounds on lactic acid concentration. Fermentations treated with antimicrobial compounds had higher final pH and lower L(+) lactic acid concentration. Narasin and salinomycin were more inhibitory than other ionophore compounds. Monensin and tylosin in combination was more effective than monensin alone. Among the nonionophore compounds, avoparcin was the least effective and thiopeptin, tylosin and virginiamycin were extremely effective in reducing lactic acid concentration. Ruminal fluid from cattle fed a diet of alfalfa hay and grain (50:50) was incubated with a mixture of sugars, casein and urea for 12 h in a buffered medium to determine the effect of antimicrobial compounds on VFA production. Generally, total VFA concentration was not affected by antimicrobial compounds except RO22-6924/004, tylosin and virginiamycin, which caused a reduction at high concentrations. Tylosin, monensin and tylosin mixture, thiopeptin and virginiamycin at high concentrations (greater than 6.0 micrograms/ml) increased the acetate proportion. All compounds increased the molar proportion of propionate. Tylosin and virginiamycin at high concentrations (greater than 6.0 micrograms/ml) decreased the proportion of propionate. Monensin and tylosin in combination had no effect on propionate proportion. Among the compounds tested, narasin and salinomycin were the most effective in enhancing propionate proportion. Ionophore compounds were more inhibitory to butyrate production than the nonionophore compounds. Batch culture fermentations may be used to quantitate the relative efficacy of antimicrobial compounds to alter ruminal fermentation characteristics.     

Effect of avilamycin, tylosin and ionophore anticoccidials on Clostridium perfringens enterotoxaemia in chickens

In order to study the prophylactic and metaphylactic effect of antomicrobial growth promoters and ionophorous anticoccidials on the incidence of Cl. perfringens enterotoxaemia in chickens, experimental attempts were performed with 675 chickens in 27 trials. The birds were intraduodenally infected with Cl. perfringens type A (ATCC 3624). The following antimicrobial growth promoters and ionophore anticoccidials were used either on their own or in combination: avilamycin, narasin, monensin and tylosin. While infected and non-medicated trials showed an average incubation period of 1 week, clinical symptoms occurred 2-4 days later in infected and medicated birds. Avilamycin medicated birds had the longest incubation period. In the infected and non-medicated trials, a mortality rate of 16%-36% was noted within 3 weeks post infection. The avilamycin trials showed a mortality rate of 0-8% (0-2 birds died) and the narasin and monensin a mortality rate of 0-8%, respectively. In the combination groups (monensin + avilamycin or narasin + avilamycin), the mortality rate ranged from 0 to 4%. Tylosin showed a very good metaphylactic/therapeutic effect against Cl. perfringens enterotoxaemia. Following infection, medicated birds showed a significantly better bodyweight gain than the chickens, whose feeds had not been supplemented. From epidemiological point of view, the systematic prevention of coccidiosis is a key in the control of Cl. perfringens enterotoxaemia in chickens.     

Susceptibility of Clostridium perfringens of animal origin to fifteen antimicrobial agents

The minimal inhibitory concentrations of fifteen antibacterial agents were determined by agar-dilution method against 121 strains of Clostridium perfringens isolated from pigs, cattle and poultry. All strains, regardless of host of origin, were susceptible to avoparcin, furazolidone, monensin, nitrovin, penicillin G, ronidazole and tiamulin and resistant to flavomycin. Poultry strains were also susceptible to carbadox, chloramphenicol, erythromycin and virginiamycin. Bacitracin-resistant poultry strains were susceptible to all tested antibacterial agents except tetracycline, but the bacitracin-resistant cattle strains were polyresistant. Porcine strains were susceptible to bacitracin and bovine strains to carbadox. Carbadox-resistant porcine strains were resistant to erythromycin, lincomycin and tetracycline and susceptible to chloramphenicol. Resistance to erythromycin was associated with resistance to lincomycin. High level erythromycin-lincomycin-resistant strains were susceptible to virginiamycin, but the intermediate level erythromycin-lincomycin-resistant cattle strains were resistant to virginiamycin. Resistance to chloramphenicol or erythromycin-lincomycin was always associated with resistance to tetracycline but the reverse was not always true.     

Bacterial flora of liver abscesses in feedlot cattle fed tylosin or no tylosin.

Bacterial flora of liver abscesses from cattle fed tylosin or no tylosin and susceptibilities of the predominant bacterial isolates to tylosin and other antimicrobial compounds were determined. Abscessed livers were collected at slaughter from cattle originating from feedlots that had fed tylosin (n = 36) or no tylosin (n = 41) for at least 2 yr, and segments of livers with one or two intact abscesses were transported to the laboratory. Abscesses were cultured for anaerobic and facultative bacteria. Fusobacterium necrophorum, either as single culture or mixed with other bacteria, was isolated from all abscesses. The incidence of subsp. necrophorum, as part of the mixed infection, was lower (P < .05) in the tylosin group than in the no-tylosin group (33 vs 61%). However, the incidence of Actinomyces pyogenes was higher (P < .01) in the tylosin group than in the no-tylosin group (53 vs 10%). Totals of 119 F. necrophorum and 21 A. pyogenes isolates were used for determinations of susceptibilities to bacitracin, oxytetracycline, chlortetracycline, lasalocid, monensin, tylosin, tilmicosin, and virginiamycin. The minimum inhibitory concentrations (MIC) of antibiotics were determined with a broth microdilution method. The mean MIC of tylosin for F. necrophorum and A. pyogenes were not different between isolates from tylosin and no-tylosin groups. We concluded that continuous feeding of tylosin did not induce resistance in F. necrophorum or A. pyogenes. Also, the higher incidence of mixed infection of F. necrophorum and A. pyogenes in liver abscesses of tylosin-fed cattle suggests a potential synergistic interaction between the two organisms in causing liver abscesses.     

The influence of dietary protein and antimicrobial feed additives on salmonella carriage by broiler chickens

Salmonella carriage in broiler chickens was not apparently influenced by the protein concentration of the ration, pelleting or by the coccidiostat monensin. The inclusion of penicillin in the diet was associated with an increase in salmonella shedding, particularly in the first half of the rearing period, but did not influence the lactobacillary count in the crop or the pH of the contents of the crop, gizzard and caecum. Furazolidone medication (150 mg/kg food) for the first 10 days had no effect on salmonella carriage at the time of slaughter (seven weeks) and studies into the influence of growth promoting antibiotics (avoparcin, bacitracin and virginiamycin) on salmonella shedding were inconclusive. Circumstantial evidence indicated that the food was the most probable source of infection for the birds studied in these experiments.     

鸡源A型产气荚膜梭菌致病性及药物疗效分析

本研究旨在调查规模化养鸡场内鸡源产气荚膜梭菌（Clostridium perfringens，CP）的致病性和耐药性，并评价阿维拉霉素和磷酸泰乐菌素预防CP感染鸡群发生坏死性肠炎（necrotizing enteritis，NE）的效果，为鸡NE的防治提供指导。在河北、山西两地选择有CP引起NE发病史的规模化养鸡场，随机采集新鲜粪便，分离CP，并通过多重PCR测定其毒素型。选定3个不同养殖场分离到的A型CP，以10⁹CFU·只^-1的剂量，连续5 d经灌胃的方式感染14日龄SPF鸡，观察肠道病变和NE发生情况，评价分离菌株的致病力。使用微量肉汤稀释法测定分离到的CP对阿维拉霉素、林可霉素和磷酸泰乐菌素的最小抑菌浓度（minimal inhibitory concentration，MIC）。根据MIC结果，选择药物敏感性较好的两种药物对因CP引起NE发病的鸡群中未出现临床症状的鸡进行预防试验，观察精神食欲、腹泻症状、肠道病变情况，统计CP检出率和NE发生率，评价两种药物预防效果。结果显示：自753份鸡粪便样品分离到91株CP，且毒素型全部为A型。攻毒试验结果显示：A、B、C组攻毒组NE病变发生率均显著高于未攻毒组D组（P<0.05），攻毒组肠道病变评分均显著高于未攻毒组（P<0.05）。与未攻毒组相比，攻毒组出现明显的腹泻症状和肠道病变，差异显著（P<0.05），3株CP都可以引起鸡NE。MIC结果表明：阿维拉霉素、林可霉素和磷酸泰乐菌素的MIC范围分别是0.25～4、0.125～128和0.25～32 μg·mL^-1。药物疗效试验结果表明：通过拌料连续21 d给予阿维拉霉素预混剂，给药期间及停药后1周可保护鸡群，防止出现NE症状或防止病变恶化，显著降低NE发生率、NE病变评分以及CP检出率（P<0.05）。通过拌料连续7 d给予磷酸泰乐菌素预混剂，效果与阿维拉霉素预混剂相当，但停药14 d后，6只鸡（6/11）又出现NE症状。目前，河北、山西省部分规模化鸡养殖场内流行的CP多为A型，并且可以引起NE，同时，A型CP对阿维拉霉素和磷酸泰乐菌素仍较敏感，其预混剂用于发病鸡群的预防效果较好。     

Antimicrobial susceptibility of Mycoplasma hyorhinis

A broth microdilution technique was used to determine the antimicrobial susceptibility of 15 field isolates of Mycoplasma hyorhinis to 10 antimicrobial agents, representative of different classes, and contrasting newer agents to existing ones. For the macrolides, the MIC(90) for tylosin and tilmicosin was 1 and 4 microg/ml, respectively, but was > or = 16 microg/ml for erythromycin. Tetracycline, lincomycin and enrofloxacin each had an MIC(90) of 2 microg/ml. The mycoplasma had similar levels of susceptibility to the aminoglycoside and aminocyclictol classes exhibiting an MIC(90) of 4 microg/ml for gentamicin and 2 microg/ml for spectinomycin. The isolates exhibited high MICs to trimethoprim/sulfamethoxazole with an MIC(90) > or = 16/304 microg/ml. In summary, M. hyorhinis isolates from the US had low MICs against a variety of antimicrobials tested, with the exception of erythromycin and trimethoprim/sulfamethoxazole.     

Antimicrobial susceptibility of Clostridium perfringens isolates of bovine, chicken, porcine, and turkey origin from Ontario

Antimicrobial susceptibilities and toxin types were determined for 275 Clostridium perfringens isolates collected in Ontario in the spring of 2005. Minimal inhibitory concentrations (MICs) of C. perfringens isolates for 12 antimicrobials used in therapy, prophylaxis, and/or growth promotion of cattle (n = 40), swine (n = 75), turkeys (n = 50), and chickens (n = 100) were determined using the microbroth dilution method. Statistical analyses and MIC distributions showed reduced susceptibility to bacitracin, clindamycin, erythromycin, florfenicol, and tetracycline for some isolates. Reduced susceptibility to bacitracin was identified in chicken (64%) and turkey (60%) isolates. Swine isolates had predominantly reduced susceptibility to clindamycin (28%) and erythromycin (31%), whereas bovine isolates had reduced susceptibility to clindamycin (10%) and florfenicol (10%). Reduced susceptibility to tetracycline was spread across all species. No clear reduced susceptibility, but elevated MIC(50) for virginiamycin was found in chicken isolates in comparison with isolates from other species. Toxin typing revealed that C. perfringens type A is the dominant toxin type isolated in this study across all 4 host species.     

Discrepancy between in vitro and in vivo assays for the susceptibility of Bacillus C.I.P. 5832 to antimicrobial agents

In vitro, avoparcin, flavomycin, virginiamycin bacitracin, trimethoprim + sulfadimethoxine, colistin, spiramycin, oxolinic acid, lincomycin + spectinomycin and a mixture of organic acids were able to inhibit Bacillus C.I.P. 5832. In vivo, none of these antibacterial agents, at the levels incorporated in feeds, could affect the viable bacillus count in the caecal content of chickens. In another in vitro assay, olaquindox, carbadox and tylosin were also effective in inhibiting Bacillus C.I.P. 5832. But, concomitantly administered with carbadox or tylosin, Bacillus C.I.P. 5832 still promoted growth and feed conversion of fattening pigs. These results, as well as data of the quoted literature, suggest that the current in vitro techniques should not be used to screen the susceptibility of Bacillus C.I.P. 5832 to antimicrobial agents. Only in vivo assays seem to be of value.     

In vitro susceptibility of 21 antimicrobial agents to 37 isolates of Brachyspira hyodysenteriae isolated from pigs in Okinawa Prefecture

The in vitro susceptibilities to 21 antimicrobial agents, of 37 isolates of Brachyspira (B.) hyodysenteriae isolated from pigs in Okinawa meat center and a pig farm in Okinawa Prefecture, Japan, were determined by the agar dilution method. Carbadox was the most active of all the agents tested against the isolates (MIC: <0.003 to 0.05). All the isolates were highly susceptible to olaquindox, tiamulin, dimetridazole, efrotomycin and valnemulin with MICs ranging from 相似文献   

Prevalence of vancomycin resistant enterococci on poultry farms established after the ban of avoparcin

Fecal samples from poultry on farms established after the ban of avoparcin (study farms) and from poultry on farms previously exposed to avoparcin (control farms) were examined for the presence of vancomycin-resistant enterococci (VRE). The samples were collected during the autumn and winter of 2001-2002. One isolate from each positive sample was selected, identified to species level, and examined for the presence of the vanA gene. The concentration of VRE and generic enterococci in the samples were also determined. In addition, the susceptibility to the ionophoric coccidiostat narasin was examined in a number of enterococcal isolates from poultry and in some enterococci of porcine origin that had not been exposed to narasin. VanA-type VRE was detected in samples from 64% of the study farms and 96% of the control farms. However, the concentration of VRE in the control samples was about six times larger than in the samples from the study farms. The minimum inhibitory concentration values for narasin differed between the poultry (1-4 mg/liter) and the porcine (0.25-0.5 mg/liter) isolates, indicating a decreased susceptibility towards narasin among enterococci from poultry.     

Drug use and antimicrobial resistance among Escherichia coli and Enterococcus spp. isolates from chicken and turkey flocks slaughtered in Quebec,Canada

An observational study was conducted of chicken and turkey flocks slaughtered at federal processing plants in the province of Quebec, Canada. The objectives were to estimate prevalence of drug use at hatchery and on farm and to identify antimicrobial resistance (AMR) in cecal Escherichia coli and Enterococcus spp. isolates and factors associated with AMR. Eighty-two chicken flocks and 59 turkey flocks were sampled. At the hatchery, the most used antimicrobial was ceftiofur in chickens (76% of flocks) and spectinomycin in turkeys (42% of flocks). Virginiamycin was the antimicrobial most frequently added to the feed in both chicken and turkey flocks. At least 1 E. coli isolate resistant to third-generation cephalosporins was present in all chicken flocks and in a third of turkey flocks. Resistance to tetracycline, streptomycin, and sulfisoxazole was detected in > 90% of flocks for E. coli isolates. Antimicrobial resistance (AMR) was observed to bacitracin, erythromycin, lincomycin, quinupristin-dalfopristin, and tetracycline in both chicken and turkey flocks for Enterococcus spp. isolates. No resistance to vancomycin was observed. The use of ceftiofur at hatchery was significantly associated with the proportion of ceftiofur-resistant E. coli isolates in chicken flocks. In turkey flocks, ceftiofur resistance was more frequent when turkeys were placed on litter previously used by chickens. Associations between drug use and resistance were observed with tetracycline (turkey) in E. coli isolates and with bacitracin (chicken and turkey), gentamicin (turkey), and tylosin (chicken) in Enterococcus spp. isolates. Further studies are needed to provide producers and veterinarians with alternative management practices and tools in order to reduce the use of antimicrobial feed additives in poultry.     

In vitro antimicrobial activity against 10 North American and European Lawsonia intracellularis isolates

The objective of this study was to determine the in vitro minimum inhibitory concentration (MIC) of antimicrobials against 10 isolates of Lawsonia intracellularis, the etiological agent of proliferative enteropathy (PE). Antimicrobials tested included carbadox, chlortetracycline, lincomycin, tiamulin, tylosin and valnemulin. The MIC of each antimicrobial against L. intracellularis was determined using a tissue culture system and was identified as the lowest concentration that inhibited 99% of L. intracellularis growth, as compared to the antimicrobial-free control. Each antimicrobial concentration was evaluated for both intracellular and extracellular activity against L. intracellularis, an obligately intracellular bacterium. When tested for intracellular activity, carbadox, tiamulin, and valnemulin were the most active antimicrobials with MICs of < or =0.5microg/ml. Tylosin (MICs ranging from 0.25 to 32microg/ml) and chlortetracycline (MICs ranging from 0.125 to 64microg/ml) showed intermediate activities and lincomycin (MICs ranging from 8 to >128mIcog/ml) showed the least activity. When tested for extracellular activity, valnemulin (MICs ranging from 0.125 to 4microg/ml) was the most active against most L. intracellularis isolates. Chlortetracycline (MICs ranging from 16 to 64microg/ml), tylosin (MICs ranging from 1 to >128microg/ml), and tiamulin (MICs ranging from 1 to 32microg/ml) showed intermediate activities. Lincomycin (MICs ranging from 32 to >128microg/ml) showed the least activity. Our in vitro results showed that each L. intracellularis isolate had a different antimicrobial sensitivity pattern and these data can be utilized as an in vitro guideline for the further antimicrobial evaluation of field L. intracellularis isolates.     

恩诺沙星与其他抗菌药对鸡毒支原体的体外联合抑菌试验

采用2倍稀释法测定了恩诺沙星及其他8种抗菌药对鸡毒支原体BG44T株的最小抑菌浓度（MIC）,再以棋盘法测定恩诺沙星分别与其他8种抗菌药不同联合对鸡毒支原体BG44T株的敏感性。结果显示：恩诺沙星、替米考星、泰乐菌素、吉他霉素、林可霉素、沃尼妙林、泰妙菌素、氯霉素及氟苯尼考对鸡毒支原体BG44T株的MIC分别为0．063、0．004、0．016、0．063、16、〈0．004、0．008、8、8μg／mL。在8种不同联合用药对鸡毒支原体BG44T株的药敏试验中,恩诺沙星＋替米考星、恩诺沙星＋泰乐菌素、恩诺沙星＋吉他霉素、恩诺沙星＋林可霉素、恩诺沙星＋沃尼妙林、恩诺沙星＋泰妙菌素联合表现出相加作用,恩诺沙星＋氯霉素、恩诺沙星＋氟苯尼考表现出拮抗作用。     

Antimicrobial susceptibility of Swedish, Norwegian and Danish isolates of Clostridium perfringens from poultry, and distribution of tetracycline resistance genes

This study was undertaken to determine the in vitro susceptibility of Clostridium perfringens, isolated from poultry to antimicrobials used in poultry production. The minimal inhibitory concentration (MIC) of eight antimicrobials, including the ionophoric coccidiostat narasin, was determined for 102 C. perfringens isolates, 58 from Sweden, 24 from Norway and 20 from Denmark. Susceptibility to each antimicrobial compound was determined by broth microdilution. The isolates were obtained from broilers (89), laying hens (9) and turkeys (4), affected by necrotic enteritis (NE) or by C. perfringens associated hepatitis (CPH), and from healthy broilers. All strains, regardless of origin, proved inherently susceptible to ampicillin, narasin, avilamycin, erythromycin and vancomycin. A low frequency of resistance to virginiamycin and bacitracin was also found. Resistance to tetracycline was found in strains isolated in all three countries; Sweden (76%), Denmark (10%) and Norway (29%). In 80% of the tetracycline-resistant isolates, the two resistance genes tetA(P) and tetB(P) were amplified by PCR whereas in 20% only the tetA(P) gene was detected. No tetM gene amplicon was obtained from any of the tetracycline-resistant isolates. The uniform susceptibility to narasin revealed in this study shows that the substance can still be used to control clostridiosis. In this study, C. perfringens also showed a low degree of resistance to most other antimicrobials tested. Despite the small amounts of tetracycline used in poultry, a considerable degree of resistance to tetracycline was found in C. perfringens isolates from Swedish broilers.     

In vitro susceptibilities of field isolates of Mycoplasma agalactiae to oxytetracycline,tylosin, enrofloxacin,spiramycin and lincomycin-spectinomycin

The minimum inhibitory concentrations (MICs) of tetracycline, enrofloxacin, tylosin, spiramycin and a lincomycin:spectinomycin 1:2 combination, against 24 Sicilian isolates of Mycoplasma agalactiae, the causative organism of contagious agalactia were determined in vitro by a broth dilution method. Enrofloxacin was the most effective antimicrobial in vitro with a range of MIC values from 0.125 to 0.500 microg/ml and an MIC(50) of 0.203 and MIC(90) of 0.365 microg/ml. Using the MIC(50) and MIC(90) values the remaining four antimicrobials are ranked in order of in vitro effectiveness as follows: tylosin (MIC(50)0.292; MIC(90)0.525 microg/ml) was slightly more effective than tetracycline (MIC(50)0.296; MIC(90)0.533 microg/ml), followed by lincomycin:spectinomycin (MIC(50)0.521; MIC(90)0.938 microg/ml) and spiramycin (MIC(50)1.583; MIC(90)2.850 microg/ml). MIC values above 1.000 microg/ml were obtained using tetracycline, tylosin and spiramycin for some M. agalactiae isolates.     

Antimicrobial susceptibility of Moraxella bovis

The antimicrobial susceptibility of 55 isolates of Moraxella bovis to seven antibiotics was evaluated by broth microdilution procedures. The isolates had an MIC₉₀ of ≤1 mg/l to erythromycin, ceftiofur, and ampicillin; 4 mg/l to tilmicosin; 16 mg/l to tylosin and gentamicin; and had MIC₉₀s of ≥32 mg/l for oxytetracycline. The modal MIC values for these antibiotics were as follows: ampicillin, <0.25 mg/l; ceftiofur, ≤0.125 mg/l; tilmicosin, 2 mg/l; tylosin, 8 mg/l; erythromycin 1 mg/l; oxytetracycline, ≤0.5 mg/l; and gentamicin, ≤0.5 mg/l. This in vitro data showed most antibiotics have low MICs that are suggestive of clinical efficacy.     

Antimicrobial susceptibility testing of Australian isolates of Brachyspira hyodysenteriae using a new broth dilution method.

The antimicrobial susceptibilities of 76 field isolates of Brachyspira hyodysenteriae from different states of Australia were tested in a newly developed broth dilution procedure. The antimicrobial agents used were tiamulin, valnemulin, tylosin, erythromycin, lincomycin and clindamycin. The results from the broth dilution susceptibility testing of 39 of the isolates were compared with results obtained for the same isolates using the agar dilution method. Amongst the isolates tested by broth dilution, 17 were from three farms and had been collected over a number of years. Their pulsed field gel electrophoresis pattern previously had been determined. The broth dilution technique was simple to use, less labor intensive than agar dilution, and gave clear end points. The results obtained using the two methods generally corresponded well, although in a few cases the MIC obtained by broth dilution were lower than those with agar dilution. For the 76 isolates tested by broth dilution, the MIC(90) (mg/l) was: tiamulin, 1; valnemulin, 0.5; tylosin>256; erythromycin>256; lincomycin, 64 and clindamycin, 16. Only minor differences in susceptibility patterns were found amongst isolates from different Australian states. Over all the isolates, and also amongst the isolates obtained from different years on the three farms, there was no trend for the susceptibility of the isolates to alter with time.     

Molecular characterization and determination of antimicrobial resistance of Mycoplasma gallisepticum isolated from chickens

In this study, three consecutive approaches of molecular characterization, determination of minimum inhibitory concentration (MIC) and antimicrobial tested on Mycoplasma gallisepticum (MG) isolated from chicken farms were investigated. These approaches were conducted between 2004 and 2005 to 134 MG samples collected from five different regions of the intensive farming area of Thailand. Twenty MG isolates and four reference strains including S6, F, ts-11, and 6/85 were classified according to Random Amplification of Polymorphic DNA (RAPD) patterns prior to the antimicrobial tests. These isolates exhibited 5 different genotypes (A-E). Consequently, MG isolates representing each genotype were tested on 11 registered antibiotics. The levels of MIC were determined. Three antibiotics, doxycycline (0.20 microg/ml), tiamulin (0.10 microg/ml), and tylosin (0.33 microg/ml), gave the least MICs among all effective drugs. Break point comparisons of each antimicrobial suggested that the MG isolates were most sensitive to lincomycin, oxytetracycline, tiamulin, and tylosin. Some MG isolates had an intermediate effect on josamycin and were resistant to enrofloxacin and erythromycin. Our results also indicated that MG isolated and collected from the region and nearby districts had similar RAPD patterns showing properties of antimicrobial resistance. The RAPD patterns may imply the frequent use of antibiotics and a resistant strain of MG. This is the first report of genetic characterization using RAPD reflected by the levels of MIC against MG. The information is useful to plan for prophylactic and therapeutic impacts on the poultry industry especially in the area of intensive use of antibiotics.