Antimicrobial susceptibility patterns of thermotolerant Campylobacter strains isolated from food animals in Ethiopia

Thermotolerant Campylobacter spp. are frequent causes of diarrhoea in humans worldwide mostly originating from poultry. It has been suggested that extensive veterinary use of antibiotics is largely responsible for resistance in human isolates. During a 4-month period from January to April 2004, 192 Campylobacter spp. were isolated from fecal samples of 485 healthy food animals. The in vitro susceptibility to 12 antibiotics was determined by the agar disk diffusion method. Among the 192 Campylobacter spp. isolated, 135 (70.3%) were identified to be C. jejuni, 51 (26.6%) were C. coli and 6 (3.1%) were C. lari. C. jejuni was the most prevalent species in chickens (80.8%) versus 16.2% C. coli and 3.0% C. lari. All isolates found in pigs were C. coli. All strains were sensitive to chloramphenicol and ciprofloxacin and all were resistant to cephalothin. More than 90% of the strains were sensitive to clindamycin, erythromycin, gentamicin, nalidixic acid, norfloxacin, streptomycin and tetracycline. Resistance was found against ampicillin in 20% and trimethoprim-sulphamethoxazole in 37.5%. Resistance was not statistically different among C. jejuni, C. coli and C. lari (p>0.05). Multidrug resistance to two or more drugs was detected in 14.5% of strains. In conclusion, the study showed that antimicrobial resistance is found only at relatively low frequencies for most antimicrobial agents tested except for ampicillin and trimethoprim-sulphamethoxazole. The low percentages of resistance to most antimicrobial agents tested in this study may be the result of low/no usage of these agents as a growth promoters or treatment in the Ethiopian animal farm setting. The detection of multidrug resistant isolates may pose a threat to humans and further limits therapeutic options.     

Antimicrobial susceptibility of Staphylococci isolated from naturally occurring canine external ocular diseases

Objective  The purpose of this study was to describe the species distribution and antimicrobial susceptibility patterns of Staphylococci isolated from naturally occurring canine ocular diseases.
Samples and Procedures  Samples obtained from 68 dogs with signs of external eye disease were processed for isolation of bacteria of the genus Staphylococcus sp. Isolates were identified by biochemical reactions and tested for susceptibility to 11 antimicrobials of six classes.
Results  Forty (58.8%) samples yielded Staphylococcus sp. in pure culture. Coagulase-positive Staphylococci were most common and Staphylococcus intermedius was the most frequent (45%) species, followed by S . aureus (22.5%) and the coagulase-negative species S. epidermidis (20%), and S . simulans (12.5%). Resistance to at least one drug was observed in 92.5% of the isolates, and multidrug resistance was a common finding (72.5%). The most effective drugs against Staphylococcus strains isolated from extra-ocular canine ocular diseases were ceftiofur and cefalexin.
Conclusion  This study highlights the presence of Staphylococcus sp. genus in naturally occurring extra-ocular canine ocular disease and the emergence of resistant strains to common antimicrobial drugs. It also emphasizes the need for bacterial culture with species identification and susceptibility testing in order to choose the appropriate antimicrobial therapy.     

Resistance of Staphylococcus-aureus strains, isolated from food, to antibiotics]

Resistance to selected antibiotics was studied in 325 strains of Staphylococcus aureus, isolated from foodstuffs. Out of these strains, 50.46% were resistant to penicillin, 15.7% to ampicillin, 4% to streptomycin, 5.2% to erythromycin, 5.8% to chloramphenicol, 18.5% to tetracycline, 18.2% to oxytetracycline, 2.5% to gentamycin, 1.6% to canamycin, 53.5% to colistine and 9.2% to bacitracin. Eighty-five strains (26.15%) were sensitive to all the antibiotics used; 107 strains (32.92%) were resistant to one antibiotic, and 133 strains (40.93%) to two or more antibiotics.     

Antimicrobial susceptibility of Staphylococcus aureus and Staphylococcus pseudintermedius isolated from various animals

This study characterized the antimicrobial susceptibility of 221 Staphylococcus aureus isolated from various species, and 60 canine Staphylococcus pseudintermedius isolated from 1986 through 2000 at the Western College of Veterinary Medicine (WCVM). Resistance of S. aureus was most common to penicillin (31%) and tetracycline (14%); resistance of S. pseudintermedius to penicillin was present in 8% and to tetracycline in 34% of isolates. Resistance to trimethoprim/sulfamethoxazole was only seen among S. pseudintermedius, and there was no resistance to amoxicillin/clavulanate, ampicillin/sulbactam, cephalothin, amikacin, gentamicin, enrofloxacin, chloramphenicol, or rifampin among any isolate. Inducible clindamycin resistance was found in both S. aureus and S. pseudintermedius, highlighting the need for careful interpretation of culture and susceptibility test results. There were significant differences in the minimum inhibitory concentrations of penicillin, ciprofloxacin, enrofloxacin, clindamycin, erythromycin, chloramphenicol, and tetracycline between avian, bovine, equine, and porcine isolates.     

兽医临床凝固酶阴性葡萄球菌分离株的耐药性及耐药基因检测

采用微量肉汤稀释法和D-试验法检测64株凝固酶阴性葡萄球菌（CNs）对10种抗菌药物的耐药性,并用PCR方法分别检测头孢西丁耐药菌株和红霉素耐药菌株中mecA基因以及erinA、ermB、ermC和msrA基因的携带情况。结果表明,所有菌株均对万古霉素和甲氧苄啶／磺胺甲恶唑敏感;泰妙菌素为耐药率（65．6％）最高的抗菌药物,其次是红霉素、氧氟沙星和β-内酰胺类药物。28株（43．8％）CNS对青霉素耐药,其中26株对头孢西丁耐药（MRS）并且均携带mecA基因。30株（46．9％）CNS对红霉素耐药,其中24株为MLSB耐药表型,主要由ermB基因介导。总之,兽医临床CNs分离株对常用抗菌药物的耐药性不同,mecA基因和ermB基因的携带分别是兽医临床MRS和MLSB表型产生的主要原因。     

广东省部分地区奶牛乳房炎凝固酶阴性葡萄球菌的毒力基因和耐药性分析

为了解广东省部分地区奶牛乳房炎凝固酶阴性葡萄球菌(coagulase negative Staphylococci,CNS)的毒力基因的分布情况和耐药现状,进而为奶牛乳房炎的防治提供依据。本试验对广东省部分地区奶牛场110份乳房炎奶样中分离出的40株CNS采用PCR方法检测毒力基因、耐药基因和耐消毒剂基因,K-B试纸片法检测药敏性。结果表明CNS毒力基因检测中检测率最高的为fnbp(17.5%),其次为seb(15%)和tsst(15%),毒力基因型复杂(15种)。耐药性检测中对链霉素、红霉素和青霉素G耐药率较高,对苯唑西林、头孢曲松和头孢唑啉较敏感,多重耐药率高(52.5%)。耐药基因检测率最高的为喹诺酮类基因qnrA/B/C/D(20%),耐药基因链霉素耐药表型主要由aac6-aph2介导,耐消毒剂基因检测率最高的为qacG(25%),耐药基因和耐消毒剂基因的基因组型复杂(14种)。     

大环内酯类抗生素对畜禽病原菌体外抗菌敏感性试验研究

针对兽医临床的细菌耐药现象,为指导临床合理用药,采用微量肉汤稀释法研究了三代大环内酯类抗生素泰乐菌素、替米考星和泰地罗新对兽医临床常见的15种共74株病原菌的体外抑菌效果。结果显示:对于革兰阳性菌,泰乐菌素和替米考星对乳房炎常见的金黄色葡萄球菌、猪链球菌、无乳链球菌、停乳链球菌均有较强的抑菌活性,优于泰地罗新;泰地罗新对肺炎链球菌抑菌效果较强。对于革兰阴性菌,如支气管败血波氏杆菌、溶血性曼氏杆菌、胸膜肺炎放线杆菌、副猪嗜血杆菌等致病菌,替米考星的抑菌效果比泰乐菌素强1～64倍,泰地罗新比替米考星强2～32倍。对引起肠道感染的大肠杆菌,泰地罗新亦表现出很好的抑菌活性,效果优于泰乐菌素和替米考星。本试验为畜禽细菌性疾病的防控以及大环内酯类抗菌药物的合理使用提供了基础数据。     

Abundance and phenotypic diversity of Escherichia coli isolates with diminished susceptibility to expanded-spectrum cephalosporins in faeces from healthy food animals after slaughter

Antimicrobial resistance (AR) is an increasing phenomenon but its quantitative estimation remains controversial. The classical resistance percentage approach is not well suited to detect either emergence or low levels resistance. One option is to shift the focus from strains to hosts. This approach is applied to test for phenotypic diversity associated with diminished susceptibility to expanded-spectrum cephalosporins (DSESC) in faecal Escherichia coli from healthy food animals in Spain.

We performed E. coli enumeration in faecal samples of broilers (82 pooled samples) and pigs (80 pooled samples) at the slaughterhouse level, using Coli-ID plates alone and supplemented with cefotaxime at two levels (1 and 8 μg/ml). Antimicrobial susceptibility of isolates was tested by the agar diffusion method. Clustering was carried out using these numerical values and Ward and UPGMA methods.

When using plates supplemented with 1 μg/ml of cefotaxime for DSESC E. coli detection, 93% (76/82) of broiler pooled samples and 36% (29/80) pig pooled samples tested positive. When using 8 μg/ml of cefotaxime, 67% (55/82) of broilers and 13% (10/80) of pigs were positive. Nevertheless, the relative abundance of this phenotype was low in both animal species (range 0–4.3%). Irrespective of the clustering method (Ward or UPGMA), a noticeable phenotypic diversity was detected, especially from the plates containing 1 μg/ml of cefotaxime.

We concluded that: (a) E. coli with phenotype DSESC are common in broilers and pigs but are less frequent in pigs, and (b) the host approach is the most appropriate method for antimicrobial resistance assessment when null or very low levels of antimicrobial resistant bacteria are expected.     

Dermatophytes isolated from domestic and feral animals

In work over a period of 8 years, dermatophytes were recovered from 12 animal species in the North Island of New Zealand. A total of 552 dermatophytes were isolated and belonged to the Microsporum (6 species) and Trichophyton (6 species) genera.

Some unusual isolations are reported: Microsporum canis and Trichophyton mentagrophytes var. mentagrophytes were recovered from calves; Microsporum distortum from a dog, which was the first known isolation from an animal in the North Island; four horses, from the same stable, yielded Microsporum equinum which has not previously been recorded in this country; Trichophyton erinacei was recovered from lesions on a cat which is the first report of the dermatophyte from an animal other than the dog, hedgehog or man; Trichophyton equinum var. autotrophicum was a rare isolation from a dog.

Species affinity was demonstrated with the dermatophytes M. canis; M. nanum; M. equinum; T. equinum; and T. verrucosum. These zoophilic species appear to be passed between individuals of the same species, with occasional infection in man and other animal species. T. mentagrophytes var. mentagrophytes had a wider distribution, being isolated from dogs, cats, guinea-pigs and rats. The infection in rats was subclinical.

The geophilic species M. cookei, T. ajelloiand and T. terrestre were recorded but not regarded as being pathogenic. M. gypseum was significant in cases involving dogs, horses and a cat, as arthrospores were seen invading the affected hairs.     

Serotypes and Shiga toxin genotypes among Escherichia coli isolated from animals and food in Argentina and Brazil

Shiga toxin (Stx)-producing Escherichia coli (STEC) strains isolated from animals and food in Argentina (n=44) and Brazil (n=20) were examined and compared in regard to their phenotypic and genotypic characteristics to evaluate their pathogenic potential. The clonal relatedness of STEC O157 isolates (n=22) was established by phage typing (PT) and pulsed-field gel electrophoresis (PFGE). All O157 strains studied carried eae and enterohemorrhagic E. coli (EHEC)-hly sequences. In Argentina, these strains occurred both in cattle and meat, and 50% of them carried stx2/stx2vh-a genes, whereas in Brazil the O157 strains were isolated from animals, and most harbored the stx2vh-a sequence. At least 13 different O:H serotypes were identified among the non-O157 strains studied, with serotype O113:H21 being found in both countries. All but one non-O157 strains did not carry eae gene, but EHEC-hlyA gene was found in 85.7% of them, and the stx2 genotype was also more prevalent in Argentina than in Brazil (P<0.01), where stx1 alone or in association was most common (68.8%). One STEC strain isolated from a calf in Brazil harbored the new variant referred to as stx2-NV206. PFGE analysis showed that STEC O157 strains were grouped in four clusters. One Brazilian strain was considered possibly related (> or =80%) to Argentinean strains of cluster I. Differences in the pathogenic potential, especially in regard to serotypes and stx genotypes, were observed among the STEC strains recovered from animals and food in both countries.     

Antimicrobial susceptibility testing of bacteria isolated from animals: methods for in-vitro susceptibility testing and their suitability with regard to the generation of the most useful data for therapeutic applications

In-vitro susceptibility testing provides valuable informations for choosing the most suitable antimicrobial agent for the control of bacterial infections in animals. Different diffusion and dilution methods, as conducted according to various approved performance standards, can be used to determine the in-vitro susceptibility of bacterial pathogens. In the present article, problems are discussed which arise from the use of different methods and the difficulty to interpret such results. While most approved performance standards were designed for testing of bacteria from human sources, the NCCLS document M31-A2 exclusively focusses on susceptibility testing of bacteria isolated from animals and--in contrast to all other standards--includes veterinary specific breakpoints for a number of antimicrobial agents used in veterinary medicine. Therefore, performance of in-vitro susceptibility testing of veterinary pathogens should follow the recommendations given in the NCCLS document M31-A2. The microdilution method is recommended as the method of choice for susceptibility testing. The result of a microdilution test is given as the minimum inhibitory concentration (MIC). This value provides a quantitative result which precisely indicates the degree of susceptibility of the tested bacterial strain and in return gives the veterinarian a clear guidance whether therapeutic intervention with the antibiotic in question will be successful.     

Comparison of minimal inhibitory concentration and disk-diffusion antimicrobic sensitivity testing of bacterial pathogens isolated from food animals

Disk-diffusion sensitivity tests were conducted with the antimicrobics sulfathiazole, gentamicin, erythromycin, kanamycin, penicillin, ampicillin, and spectinomycin on 300 to 350 bacterial isolates of food animal origin. The minimal inhibitory concentration of each antimicrobic was also determined for each bacterial isolate, using a microdilution technique. Results indicated that inhibitory zone sizes should be larger for some antimicrobics when testing animal pathogens than those zone sizes recommended for testing human pathogens. In addition, zone interpretive data are reported for spectinomycin, a drug for which such data were previously lacking.     

Susceptibility of different bacterial species isolated from food animals to copper sulphate, zinc chloride and antimicrobial substances used for disinfection

A total of 569 different bacterial isolates (156 Salmonella, 202 E. coli, 43 S. aureus, 38 S. hyicus, 52 E. faecalis, 78 E. faecium) were tested for susceptibility to copper sulphate, benzalkonium chloride, hydrogen peroxide and chlorhexidine using MIC determinations. A total of 442 isolates were also tested for susceptibility to formaldehyde and 177 isolates for susceptibility to zinc chloride. Enterococcal isolates formed a bimodal distribution of MICs to copper sulphate, whereas the other bacterial species formed one large population. Otherwise the isolates formed one large population of susceptibilities to the different antimicrobial agents. Large variations were observed in the susceptibility of the different bacterial species to the different compounds. Staphylococci were in general very susceptible to all antimicrobial compounds tested. The Salmonella isolates were in general less susceptible to copper sulphate, benzalkonium chloride and chlorhexidine followed by E. coli and the Gram-positive species. The opposite was the case for zinc chloride. All isolates were very susceptible to H(2)O(2) with MICs ranging from 0.002 to 0.016%, and to formaldehyde with MICs at 0.003 and 0.006%. This study showed that Danish bacterial isolates from livestock so far have not or have only to a limited degree developed resistance to antimicrobial compounds commonly used for disinfection. Acquired copper resistance was only found in enterococci. There were large differences in the intrinsic susceptibility of the different bacterial species to these compounds, and Salmonella especially seems intrinsically less susceptible than the other bacterial species, which might have human health implications.