Genetic diversity and tetracycline resistance genes of Histophilus somni

Nasal swabs were collected at three time points from 2378 calves in four feedlots and cultured for Histophilus somni to assess genetic relatedness and tetracycline resistance. The proportions of animals carrying tetracycline resistant isolates were 0.32% at arrival, 14.82% at interim, and 0.80% at exit. The 606 H. somni isolates recovered were compared by pulsed-field gel electrophoresis (PFGE), screened for the presence of plasmids, and assessed for the tetracycline resistance genes tet(A), tet(B), tet(C), tet(E), tet(G), tet(H), tet(K), tet(L), tet(M) and tet(O) using multiplex polymerase chain reaction. Most of the isolates (98.6%) belonged to one of seven PFGE clusters (A-G) of closely related profiles with 77.7% of the isolates belonging to clusters C and D. Clusters A, B and E were associated with a higher proportion of tetracycline susceptible isolates. Genetic diversity of the isolates was highest at entry in the feedlot and lowest after the period when the animals received in-feed chlortetracycline (interim samples). Clusters A and E were more prominently represented at exit from the feedlot than other clusters. All resistant strains harboured the gene tet(H) while no other tetracycline resistance genes and no plasmids were detected with the methodology employed. It appears that genetic variability in H. somni in Alberta feedlots is low, dissemination likely occurs by clonal expansion, and resistance to tetracyclines is mediated by the tet(H) encoded efflux pump. Pulsotypes associated with tetracycline susceptible strains appear more common at exit suggesting that the in-feed oxytetracycline included throughout the feeding period is not sufficient to exert selective pressure for resistant strains.     

Antimicrobial susceptibility and presence of resistance genes in staphylococci from poultry

The species distribution, susceptibility to 19 antimicrobial agents and presence of selected genes encoding resistance to macrolides, streptogramins and tetracyclines were examined among 118 staphylococcal isolates from infections of poultry in Denmark. Isolates were identified using a combination of conventional biochemical testing and 16S rDNA sequencing. The most common species were Staphylococcus aureus (83), Staphylococcus hyicus (11), Staphylococcus xylosus (9) and Staphylococcus cohnii (6). The isolates were susceptible to most antimicrobials tested. A high frequency of S. aureus (30%) was resistant to ciprofloxacin. Only six (7%) S. aureus isolates and one Staphylococcus saprophyticus were penicillin resistant. Resistance to sulphamethoxazole was observed among 16 (19%) of S. aureus isolates and two coagulase negative staphylococci (CNS). Twenty (24%) of the S. aureus isolates were resistant to erythromycin and 19 of these isolates contained the ermA gene, whereas the remaining isolate contained the ermC gene. Eleven (48%) of the novobiocin resistant CNS were resistant to erythromycin and all these isolates contained the ermA gene. Two isolates identified as S. xylosus, were found to be resistant to streptogramins and both contained the vatB- and the vgaB-genes. Thirty-nine (47%) of the S. aureus isolates, three of nine S. hyicus and eight of the 23 novobiocin resistant CNS were tetracycline resistant and all contained the tet(K) gene. A single S. aureus isolate also contained the tet(M) gene. The present study showed a frequent occurrence of resistance to fluoroquinolones, tetracycline and macrolides among staphylococci isolated from broilers in Denmark, whereas the occurrence of resistance to other antimicrobial agents remains low. Similar genes, encoding resistance to erythromycin, tetracycline and streptogramins to those previously observed, were detected.     

Characterization of Streptococcus suis serotype 7 isolates from diseased pigs in Denmark

Isolates of Streptococcus suis serotype 7 from diseased pigs in Denmark were characterized by ribotyping, pulsed field gel electrophoresis (PFGE), MIC-determinations and detection of resistance genes. Forty-one different ribotype profiles were found among the 103 isolates and could be divided into two main clusters. No obvious relationship between ribotypes and the clinical origin of the isolates could be observed. Fifty-four isolates, including all 24 isolates belonging to the main ribotype profile were examined by PFGE and 50 different profiles were found. A high frequency of resistance to erythromycin (41%), tetracycline (24%) and streptomycin (28%) was observed. Furthermore, almost all isolates (101) were resistant to sulphamethoxazol. Most isolates were susceptible to ceftiofur, chloramphenicol, florfenicol, penicillin, ciprofloxacin, trimethoprim and trimethoprim + sulphonamides. The tet(M) gene was found among 11 (44%) and the tet(O) gene in six (24%) of 25 tetracycline resistant isolates. The tet(L) and tet(S) genes were not detected in any isolates. The erm(B) gene was detected in 39 (93%) of 42 erythromycin resistant isolates.     

Antimicrobial susceptibilities of thermophilic Campylobacter from humans, swine, and chicken broilers

The aim of this study was to evaluate the incidence and the distribution of antimicrobial resistance, and the presence of genetic determinants of resistance, in Campylobacter recovered from swine, poultry, and human populations in Quebec. Minimum inhibitory concentrations (MICs) of 10 antimicrobial agents were determined by the agar dilution technique. Polymerase chain reaction (PCR) was used to detect the tetO determinant, and mutations in gyrA were analyzed by sequencing and by mismatch amplification mutation assay (MAMA) PCR. Among C. coli isolates from pigs, the rates of resistance were high, at 59% for clindamycin, 61% for erythromycin, 67% for streptomycin, and 68% for tetracycline; isolates from chicken broilers were mainly resistant to streptomycin and tetracycline, with a rate of 50% for each; and 56% of the isolates from humans were resistant to tetracycline. The rates of resistance among C. jejuni isolates were low except for tetracycline (39% and 67% in humans and broilers, respectively). The tetO determinant was identified among both tetracycline-resistant and tetracycline-susceptible Campylobacter isolates from swine. Sequencing analysis showed that 64% and 100% of ciprofloxacin-resistant C. coli isolates from swine and humans, respectively, had the mutation Thr-86-->Ile, which is associated with quinolone resistance. The MAMA PCR gave identical results. Further analyses need to be done in order to detect other genetic determinants of tetracycline resistance.     

禽源沙门菌对四环素的耐药性及耐药基因调查分析

为了更好地了解禽源沙门菌对四环素的耐药性及耐药基因分布,从不同来源的家禽样品中分离沙门菌,调查其对四环素的耐药性以及耐药菌株中8种四环素耐药基因[tet(A)、tet(B)、tet(C)、tet(W)、tet(M)、tet(D)、tet(K)和tet(L)]的携带情况.结果表明,18.8%的沙门菌分离株对四环素耐药,健康成鸡分离株对四环素的耐药性明显高于雏鸡、死胚或病禽分离株;四环素耐药株中tet(A)、tet(B)和tet(M)基因的携带比例分别为73.1%、11.5%和3.8%,说明沙门菌对四环素的耐药机制以tet(A)和tet(B)基因介导的主动外排为主.本研究首次在对四环素耐药的沙门菌中检测到tet(M)基因,说明tet(M)基因在沙门菌对四环素的耐药性方面也具有潜在的作用.     

Antimicrobial drug resistance as determined by the E-test in Campylobacter jejuni, C. coli, and C. lari isolates from the ceca of broiler and layer chickens in Grenada

One hundred and twenty five chickens from Grenada, consisting of 77 broilers and 48 layers were examined for carriage of thermophilic campylobacters in their ceca by culture. Seventy nine percent of chickens were positive for campylobacters, with an isolation rate of 93.5% for broilers and 56.3% for layers, the difference being significant. Sixty-four pure cultures comprising 39 Campylobacter coli, 21 Campylobacter jejuni, and 4 Campyilobacter lari isolates were tested for their resistance against 7 antibiotics using the E-test. None of the isolates were resistant to chloramphenicol and gentamicin. Resistance rates to other drugs were: ampicillin, 9.4%; ciprofloxacin, 12.5%; erythromycin, 3.1%; metronidazole, 9.4%, and tetracycline, 50% with MICs of >or=256 microg/mL for tetracycline. There were no significant differences in resistance rates between C. coli and C. jejuni. Multiple resistance to >or=2 drugs was seen in 15.6% of total isolates. All C. lari isolates were resistant to ciprofloxacin, and 3 of 4 isolates had multiple drug resistance. Overall, erythromycin, which is the drug of choice for treatment of Campylobacter infections in humans, is effective in vitro against 97% of chicken isolates in Grenada.     

Evaluation of antimicrobial susceptibility patterns in Campylobacter spp isolated from dairy cattle and farms managed organically and conventionally in the midwestern and northeastern United States

OBJECTIVE: To describe antimicrobial susceptibility patterns in Campylobacter spp isolated from dairy cattle and farms managed organically and conventionally in the midwestern and northeastern United States. DESIGN: Longitudinal study. SAMPLE POPULATION: 128 farms. PROCEDURE: Samples and data were collected every 2 months from August 2000 to October 2001. Fecal samples were collected from calves and cows. Milk samples were obtained from the bulk tank and milk line filters. Environmental samples were obtained from a water source, feed bunks of lactating cows, and cattle housing areas. Campylobacter identification and antimicrobial susceptibility testing were performed at a central laboratory by use of microbroth dilution with 2 customized antimicrobial susceptibility panels. RESULTS: 460 and 1,570 Campylobacter isolates were obtained from organic and conventional dairy farms, respectively. Most isolates from both farm types were susceptible to most antimicrobial agents tested, and antimicrobial susceptibility of conventional dairy isolates was decreased, compared with organic dairy isolates. Low proportions of isolates resistant to ampicillin (< 10%) and moderate proportions resistant (30% to 60%) to kanamycin, sulfamethoxazole, and tetracycline were observed on both farm types. The proportion of isolates resistant to tetracycline was higher for conventional than organic farms. CONCLUSIONS AND CLINICAL RELEVANCE: Campylobacter isolates from dairy cattle and farms managed organically and conventionally had similar patterns of antimicrobial resistance; the proportion of resistant isolates was higher for conventional than organic farms.     

Identification of the tetracycline resistance gene, tet(M), in Erysipelothrix rhusiopathiae

This is the first report to demonstrate the presence of tet(M) in naturally occurring isolates of tetracycline-resistant Erysipelothrix rbusiopathiae, which causes swine erysipelas. The tet(M) gene was isolated from E. rhusiopathiae strain KY5-42. The nucleotide and the deduced amino acid sequence were 99% identical to the tet(M) gene from Enterococcus faecalis. The gene was necessary and sufficient for the expression of tetracycline resistance in Escherichia coli. The presence of the tet(M) gene in the 114 tetracycline-resistant E. rhusiopathiae isolates from diseased pigs was detected by the polymerase chain reaction assay. The specific amplified DNA fragment was obtained from all 114 tetracycline-resistant strains. It was suggested that the tet(M) gene was widely present in the field isolates of E. rhusiopathiae resistant to tetracycline.     

Multiple typing for the epidemiological study of contamination of broilers with thermotolerant Campylobacter

This study aims to investigate the genetic diversity of thermotolerant Campylobacter in commercial broiler flocks and in the environment of broiler farms in Belgium. Seven out of 18 investigated flocks became colonized during rearing. Fluorescent amplified fragment length polymorphism (FAFLP), pulsed field gel electrophoresis (PFGE), restriction fragment length polymorphism of the flagellin A gene (flaA-RFLP) and antimicrobial resistance profile (ARP) were used for typing of the isolates. By the combination of FAFLP and PFGE, 22 Campylobacter genotypes could be distinguished. Colonization was almost exclusively with Campylobacter jejuni and unique genotypes were found in each flock. Multiple genotypes were detected in the broilers of 3 flocks, either simultaneously or successively. In 5 flocks, strains that were resistant to at least one antibiotic (mostly tetracycline) were found. The presence of other broiler houses on the farm did not result in a higher probability of colonization. The nipple water was contaminated with the same genotype as the broilers, illustrating its importance for transmission of Campylobacter. The same genotype was detected in a water puddle and in the broiler flock during rearing in 3 flocks. Once, the same genotype was isolated from the ditch water shortly before it was detected in the broilers.     

The rarely reported tet(31) tetracycline resistance determinant is common in Gallibacterium anatis

The present investigation was undertaken to identify and characterize the tetracycline resistance determinant in 22 Gallibacterium anatis strains for which no determinant was identified using primers specific for tet(A, B, C, D, E, G, H, K, L, M, O). A recent study found tet(B) to be the most prevalent tetracycline resistance determinant in a larger collection of G. anatis field strains from Mexico and Denmark. However, in 41% of the tetracycline resistant strains no determinant could be assigned. Here we demonstrate that tet(31) is a common determinant in G. anatis originating from chickens from very different production systems and localities. In addition, tet(31) was identified in strains isolated over a 30-year period. This is the first report on tet(31) since its original identification in Aeromonas salmonicida.     

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.     

Antimicrobial resistance, virulence-associated genes, and pulsed-field gel electrophoresis profiles of Salmonella enterica subsp. enterica serovar Typhimurium isolated from piglets with diarrhea in Korea

Salmonella enterica subsp. enterica serovar Typhimurium was isolated from diarrheic piglets in 2 periods, 2000-2001 (n = 25) and 2005-2006 (n = 17). To compare the characteristics of the isolates collected during the 2 periods, all isolates were tested for antimicrobial resistance, the presence of virulence genes, and pulsed-field gel electrophoresis (PFGE) patterns. All 42 isolates were resistant to at least 1 of the 20 antimicrobials tested, and 39 (93%) were resistant to 2 or more antimicrobials. One isolate was resistant to 12 antimicrobials. Profiles of antimicrobial resistance revealed 20 resistance types. Several isolates were also resistant to quinolones and expanded-spectrum cephalosporins. Ten isolates (24%) were resistant to ampicillin, chloramphenicol, streptomycin, sulfonamides, and tetracycline (ACSSuT); only one isolate had been isolated in 2000-2001, indicating that this type of resistance has rapidly disseminated. Polymerase chain reaction (PCR) assays revealed that all the isolates carried invA. Among the 25 strains isolated in 2000-2001, all carried the sipA, sopA, sopD, sopE2, and ssaR genes, and 96% carried sopB and sifA. Among the 17 strains isolated in 2005-2006, all carried sifA, and approximately 90% carried sipA, sopA, sopB, sopD, sopE2, and ssaR. However, only 6 (14%) of the 42 isolates carried spvC. By PFGE analysis, all 42 strains were classified into 4 major clusters, basically by collection period. The genetic similarity according to PFGE suggests that the strains isolated from diarrheic piglets of this region within the same period may be closely related.     

Antimicrobial resistance in Campylobacter from broilers: association with production type and antimicrobial use

The isolation and antimicrobial resistance of Campylobacter jejuni and Campylobacter coli strains from broilers arriving in French slaughterhouses, were analysed according to production types (i.e. standard, export or free-range) and antimicrobial (i.e. coccidiostats, growth promoters or therapeutic agents) administration in flocks. Prevalence was 56.6% in standard, 51.3% in export and 80.0% in free-range broilers. Three hundred and ninety-three strains were identified. Two-thirds of the strains belonged to the species C. jejuni. The others were C. coli. Antimicrobial susceptibility testing was carried out for ampicillin, nalidixic acid, enrofloxacin, tetracycline, erythromycin and gentamicin according to a dilution method. The percentages of resistant strains were, 23, 25, 17, 57, 0.3 and 0% for C. jejuni and 29, 43, 40, 70, 31 and 0% for C. coli. Statistical analysis revealed significant difference in distribution of C. jejuni and C. coli and antimicrobial resistance according to production type or antimicrobial administration.     

Phenotypic and genotypic differentiation of porcine Salmonella enterica subsp. enterica serovar Derby isolates

Sixty-two Salmonella enterica subsp. enterica serovar Derby isolates from slaughter pigs and meat products isolated in Southern Brazil were analyzed for their genomic relationships and for the presence of antimicrobial resistance genes. Twenty-four S. Derby isolates were indistinguishable by their subtracted restriction fingerprinting (SRF) pattern, XbaI- and BlnI-macrorestriction patterns, phage type, plasmid profile, and resistance pattern. In contrast to the BlnI-macrorestriction patterns, the XbaI-macrorestriction patterns were in good agreement with the results of SRF analysis and phage typing. Among the four phage types detected, PT10 and PT21 were the most common. The combination of all typing methods revealed a great diversity among the S. Derby isolates. All strains carried plasmids and the 60 resistant isolates showed at least tetracycline resistance. The resistance genes found were sul1 and/or sul2 (sulfonamide resistance), aadA2 (streptomycin/spectinomycin resistance), tet(A) (tetracycline resistance), tet(B) (tetracycline/minocycline resistance), bla(TEM) (ampicillin resistance), and dfrA14 (trimethoprim resistance). A correlation of the geno- and phenotypic characteristics with the origin of the isolates revealed a substantial temporal variation in the occurrence of specific S. Derby isolates in different independent pig production lines in Southern Brazil. The large number of resistant isolates underlined the potential risk that S. Derby isolates can pose to human health when they enter the food chain.     

Susceptibility of Escherichia coli and Enterococcus faecium isolated from pigs and broiler chickens to tetracycline degradation products and distribution of tetracycline resistance determinants in E. coli from food animals

One hundred Escherichia coli isolates from diseased and healthy pigs, cattle and broiler chickens were screened for the presence of tetracycline resistance genes tet(A), (B), (C), (D) or (E). The tet(A) gene was the most abundant (71% of the 100 isolates) followed by tet(B) (25%). The predominance of tet(A) and tet(B) applied to all three animal species, and there was no difference between the distribution of tet(A) and tet(B) genes among non-pathogenic and pathogenic E. coli in any of the animal species. The susceptibility of 20 of these isolates together with 10 tetracycline sensitive E. coli and 18 tetracycline resistant and 10 sensitive Enterococcus faecium to tetracyclines and tetracycline degradation products was determined. The resistant isolates showed reduced resistance to anhydrotetracycline, 4-epi-anhydrotetracycline, anhydrochlortetracycline and 4-epi-anhydrochlortetracycline. In general both the tetracycline resistant and susceptible E. faecium were more susceptible to the compounds tested than E. coli.     

Phage types, ribotypes and tetracycline resistance genes of Salmonella enterica subsp. enterica serovar Typhimurium strains isolated from different origins in Italy

The tetracycline resistance (tet) gene patterns of 52 tetracycline resistant Salmonella enterica subsp. enterica (S.) serovar Typhimurium isolates collected from animals, food of animal origin, and humans in Italy, were investigated to evaluate whether the tet gene patterns could be used for strain differentiation in addition to phage typing and ribotyping. The detection of tet genes was performed by specific PCR assays. Ribotyping was performed automatically using PvuII as restriction enzyme. Ten different ribotyping patterns were detected. All isolates were positive for at least one of the tet genes studied and six different tet gene patterns were observed. Ribotyping and tet gene patterns showed discriminatory indices of 0.741 and 0.812, respectively. Multiple tet genes were commonly found among tetracycline resistant S. typhimurium isolates from various sources. The resulting tet gene patterns allowed further discrimination of strains which were otherwise indistinguishable by their phage type, ribotype and origin. Thus, the analysis of tet gene patterns might represent an additional tool for the differentiation of S. typhimurium isolates.     

Prevalence and Genetic Relatedness of Antimicrobial‐Resistant Escherichia coli Isolated From Animals,Foods and Humans in Iceland

The prevalence of resistant bacteria in food products in Iceland is unknown, and little is known of the prevalence in production animals. The aim of this study was to investigate the prevalence and genetic relatedness of antimicrobial‐resistant Escherichia coli from healthy pigs and broiler chicken, pork, broiler meat, slaughterhouse personnel and outpatients in Iceland. A total of 419 E. coli isolates were tested for antimicrobial susceptibility using a microbroth dilution method (VetMIC), and resistant strains were compared using pulsed‐field gel electrophoresis (PFGE). All samples were screened for enrofloxacin‐resistant strains with selective agar plates. The resistance rates among E. coli isolates were moderate to high from caecal and meat samples of pigs (54.1% and 28%), broilers (33.6% and 52%) and slaughterhouse personnel (39.1%), whereas isolates from outpatients showed moderate resistance rates (23.1%). Of notice was resistance to quinolones (minimum inhibitory concentrations: nalidixic acid ≥ 32, ciprofloxacin ≥ 0.12 and enrofloxacin ≥ 0.5), particularly among broiler and broiler meat isolates (18.2% and 36%), as there is no known antimicrobial selection pressure in the broiler production in Iceland. The majority (78.6%) of the resistant E. coli isolates was genotypically different, based on PFGE fingerprint analyses and clustering was limited. However, the same resistance pattern and pulsotype were found among isolates from broiler meat and a slaughterhouse worker, indicating spread of antimicrobial‐resistant E. coli from animals to humans. Diverse resistance patterns and pulsotypes suggest the presence of a large population of resistant E. coli in production animals in Iceland. This study gives baseline information on the prevalence of antimicrobial‐resistant E. coli from production animals, and their food products in Iceland and the moderate to high resistance rates emphasize the need for continuing surveillance. Further studies on the origin of the resistant strains and the genetic relatedness of strains of different origin are needed.     

Evidence for plasmid-mediated tetracycline resistance in Paenibacillus larvae, the causal agent of American Foulbrood (AFB) disease in honeybees

Paenibacillus larvae is the causal agent of American Foulbrood (AFB) disease, the most virulent bacterial disease of honeybee (Apis mellifera L.) brood. Oxytetracycline is the main antibiotic used for prevention and control of AFB. Using the polymerase chain reaction, isolates were screened for the presence of the tetracycline resistance tet(K) and tet(L) determinants. Four isolates (5%), which correlated with the Tc-resistant phenotypes, were found to carry the tet(K) determinant, whereas none carried the tet(L) determinant. P. larvae cells were also screened for the presence of extrachromosomal DNA and evidence obtained that tetracycline resistance is plasmid-encoded. A few P. larvae isolates were found to be able to transfer the tet(K) determinant to Bacillus subtilis, suggesting that a conjugation mechanism may be involved in the transfer of the tetracycline-resistant phenotype. Minimum inhibitory concentrations to tetracycline were determined for 75 isolates of P. larvae from different geographical origins and found to range between 0.062 and 128 microg tetracyclineml(-1), with MIC(50) and MIC(90) values of 1 and 4, respectively. According to results from P. larvae populations, isolates could be considered as susceptible when their MICs were <4, intermediate for MICs values 4-8 and resistant for MICs > or = 16. To our knowledge, this is the first report of Tc(r)Paenibacillus species carrying a tet(K) gene, and also the first record of P. larvae strains carrying tet(K) determinants and its correlation with the presence of extrachromosomal DNA.     

Trends in antimicrobial susceptibility among isolates of Campylobacter species in Ireland and the emergence of resistance to ciprofloxacin

Measurements were made of the susceptibility to six commonly prescribed antibiotics, including erythromycin, tetracycline and ciprofloxacin, of 130 isolates of Campylobacterjejuni and 15 isolates of Campylobacter coli cultured from human and poultry sources during 2000. The results were compared with the results from a collection of strains isolated between 1996 and 1998. The levels of resistance to erythromycin remained low, 2 per cent and 4.4 per cent for the human and poultry isolates, respectively. Resistance to tetracycline had increased to 31 per cent and 24.4 per cent from 13.9 per cent and 18.8 per cent for the human and poultry isolates, respectively. However, the resistance to ciprofloxacin of the strains isolated during 2000 had increased to 30 per cent, whereas between 1996 and 1998 there had been no resistance to this agent among human isolates, and only 3.1 per cent resistance among poultry isolates. The molecular basis for this resistance has been shown to be the result of a single amino acid substitution, Thr-86-Ile, in the gyrA subunit of DNA gyrase in Cjejuni. A subset of 59 isolates was tested by molecular methods and all of the 25 phenotypically resistant isolates possessed this substitution. None of the human isolates had been treated with ciprofloxacin before their laboratory isolation.     

Antimicrobial resistance in Clostridium perfringens isolates from broilers in Belgium

The antimicrobial susceptibility of Clostridium perfringens strains isolated from Belgian broilers between May and September 2007 was investigated. All 39 tested isolates were sensitive to enrofloxacin, erythromycin, tylosin, florfenicol and bacitracin. Twenty-six (66%) and 24 (61%) out of the 39 tested isolates showed acquired resistance to tetracycline and lincomycin, respectively. The C. perfringens isolates were also screened by PCR for the presence of the resistance genes tet(K), tet(L), tet(M), tetB(P), tet(O), tet(W), lnu(A) and lnu(B). In 22/26 tetracycline resistant strains and 7/24 lincomycin resistant strains, resistance could be attributed to one or more of these genes. An extended frequency distribution range of MICs was seen for ampicillin. These data are consistent with data derived from studies carried out in 1980 and in 2004, indicating that no changes in antimicrobial resistance patterns have taken place during time in C. perfringens isolates from broilers in Belgium.