Trends in fluoroquinolone resistance of bacteria isolated from canine urinary tracts.

Fluoroquinolone (FQ) antimicrobial agents are used extensively in human and veterinary medicine. Widespread use of any antimicrobial agent can apply selective pressure on populations of bacteria, which may result in an increase in the prevalence of antimicrobial-resistant isolates. Antimicrobial-susceptibility data on bacteria isolated from the canine urinary tract by the University of Missouri-Columbia Veterinary Medical Diagnostic Laboratory, Columbia, MO, were used to determine whether there has been an increase in the prevalence of FQ-resistant bacteria over time. Between January 1992 and December 2001, minimum inhibitory concentrations of either ciprofloxacin (1992-1998) or enrofloxacin (1998-2001) were determined for 1,478 bacterial isolates from the canine urinary tract. The predominant bacterial species isolated were Escherichia coli (547 isolates), Proteus mirabilis (156), and Staphylococcus intermedius (147). In all, there were 13 bacterial species with more than 25 isolates each. A significant increase in the overall proportion of resistant bacterial isolates was documented from 1992 to 2001 (Cochran-Armitage test for trend, P < 0.0001). The same increase in resistant isolates was documented when either ciprofloxacin or enrofloxacin was analyzed separately (P < 0.0001 and P < 0.0002, respectively). No difference was detected in rates of bacterial FQ resistance with regard to the sex of the dog from which the bacteria were isolated. The frequency with which some bacterial species were isolated differed with the sex of the infected dog. Proteus mirabilis was found more often in females (P < 0.0001), whereas beta hemolytic Streptococcus spp., were found more often in males (P = 0.0003). Although the overall efficacy of FQ antimicrobials remained high with greater than 80% of isolates being susceptible, the data demonstrated an increase in the proportion of resistant bacteria isolated from the urinary tract of the dog.     

Antimicrobial susceptibility of Pseudomonas aeruginosa isolates from dogs with otitis externa

Pseudomonas aeruginosa is a common cause of otitis externa in dogs, and treatment of these infections is becoming problematic because of the increasing number of multiresistant strains. The aim of the present study was to compare the in vitro activities of cefepime, ceftazidime, enrofloxacin, ciprofloxacin, gentamicin and ticarcillin/clavulanic acid against 104 strains of P aeruginosa isolated from dogs with otitis externa. Antimicrobial susceptibility and minimum inhibitory concentrations, in μg/ml, were evaluated by the E test (bioMérieux). The most active compound was ceftazidime, with 100 per cent efficiency. The majority of tested strains were susceptible to ticarcillin/clavulanic acid (89.4 per cent), followed by ciprofloxacin (88.5 per cent) and cefepime (60.6 per cent). The highest resistance was observed to enrofloxacin (51.9 per cent) and gentamicin (43.3 per cent). Large numbers of strains were intermediately susceptible to antibiotics registered for use in veterinary medicine in Croatia--enrofloxacin (47.1 per cent) and gentamicin (41.3 per cent).     

Antimicrobial resistance of Escherichia coli isolates from canine urinary tract infections

This study determined the antimicrobial resistance profiles of Escherichia coli isolates from dogs with a presumptive diagnosis of urinary tract infection (UTI). Urine samples from 201 dogs with UTI diagnosed through clinical examination and urinalysis were processed for isolation of Escherichia coli. Colonies from pure cultures were identified by biochemical reactions (n=114) and were tested for susceptibility to 18 antimicrobials. The two most frequent antimicrobials showing resistance in Urinary E. coli isolates were oxytetracycline and ampicillin. Among the resistant isolates, 17 resistance patterns were observed, with 12 patterns involving multidrug resistance (MDR). Of the 69 tetracycline-resistant E. coli isolates, tet(B) was the predominant resistance determinant and was detected in 50.9% of the isolates, whereas the remaining 25.5% isolates carried the tet(A) determinant. Most ampicillin and/or amoxicillin-resistant E. coli isolates carried bla_TEM-1 genes. Class 1 integrons were prevalent (28.9%) and contained previously described gene cassettes that are implicated primarily in resistance to aminoglycosides and trimethoprim (dfrA1, dfrA17-aadA5). Of the 44 quinolone-resistant E. coli isolates, 38 were resistant to nalidixic acid, and 6 were resistant to nalidixic acid, ciprofloxacin and enrofloxacin. Chromosomal point mutations were found in the GyrA (Ser83Leu) and ParC (Ser80Ile) genes. Furthermore, the aminoglycoside resistance gene aacC2, the chloramphenicol resistant gene cmlA and the florfenicol resistant gene floR were also identified. This study revealed an alarming rate of antimicrobial resistance among E. coli isolates from dogs with UTIs.     

Antimicrobial resistance of Actinobacillus pleuropneumoniae isolated from swine

The aim of this retrospective study was to evaluate the antimicrobial resistance rates and the trend in resistance of Actinobacillus pleuropneumoniae isolated from pigs in Italy from 1994 to 2009. A total of 992 A. pleuropneumoniae isolates were tested for their susceptibility to a panel of antimicrobial agents in a disk diffusion method. Resistance to 7 drugs (amoxicillin, amoxicillin/clavulanic acid, ampicillin, cefquinome, cotrimoxazole, penicillin G and tilmicosin) showed a significant increasing trend over the time, while for 2 drugs (gentamycin and marbofloxacin) a significant decrease was observed. Resistance to the remaining 14 antimicrobial agents tested did not change significantly over the study period. Most of the isolates retained high susceptibility to antimicrobials usually effective against A. pleuropneumoniae such as amphenicols, fluoroquinolones and ceftiofur. However, high rates of resistance were observed for potentiated sulfa drugs, tetracyclines and penicillins which are currently recommended antimicrobials for pig pleuropneumonia therapy. Our results suggest the importance of continued monitoring of A. pleuropneumoniae clinical isolates in order to choose the most appropriate treatment of infections and to control the increase of resistance to currently used antimicrobials.     

水貂绿脓杆菌分离株的生物学特性和血清型分析

为了解国内水貂出血性肺炎病原绿脓杆菌的生物学特性和血清型分布,2002—2010年间从国内5个省水貂养殖场暴发出血性肺炎的水貂肺中分离获得45株绿脓杆菌,测定了分离株的生化特性、血清型、抗生素敏感性和对小鼠和水貂的半数致死量(LD50)。结果表明:分离菌株的主要生化指标除甘露醇和ONPG与人医临床分离株的不同外,其余均与绿脓杆菌的生理生化特性相同。45株菌共分为3种血清型:G型(42/45),B型(2/45)和I型(1/45),此结果与国外近几十年流行的水貂出血性肺炎和人医临床分离的绿脓杆菌主要流行血清型相同。45株菌对5种抗生素均较敏感,敏感性与分离的年份和菌株的血清型无关系。经动物试验证明,所选的4株菌毒力均不同,2株B型菌株毒力均较G型弱,4菌株对水貂的毒力均强于对小鼠的毒力,说明菌株对水貂较易感。     

Antimicrobial susceptibility of Prototheca zopfii isolated from bovine intramammary infections

In vitro antimicrobial susceptibility tests were carried out on 48 strains of Prototheca zopfii, an achlorophyllous algae causing refractory mastitis in dairy cows; 27 antimicrobials were evaluated. All strains were susceptible to both myxin and nystatin. In addition, 22 strains were susceptible to amphotericin B, 21 to polymyxin B, and 18 to gentamicin. Only 1 strain was susceptible to kanamycin. All strains were resistant to ampicillin, bacitracin, carbenicillin, cephalothin, chloramphenicol, clotrimazole, cloxacillin, erythromycin, flucytosine, ketoconazole, lincomycin, miconazole, neomycin, nitrofurazone, novobiocin, oleandomycin, penicillin, rifampin, streptomycin, tetracycline, and vancomycin.     

Antimicrobial activity of enrofloxacin against Staphylococcus intermedius strains isolated from canine pyodermas

This study examined and compared the minimal inhibition concentrations (MICs) of enrofloxacin against 393 Staphylococcus intermedius strains isolated in France from canine pyodermas during three different years, 1995 (174 isolates), 1997 (101 isolates) and 1999 (118 isolates). The MICs of enrofloxacin against these strains ranged from 0.063 to 64 mg L^?1, with MIC₅₀ and MIC₉₀ equal to 0.125 and 0.25 mg L^?1, respectively. Two resistant strains were found, but only among isolates collected in 1999. The data show that resistance to enrofloxacin among S. intermedius strains is still rare in dogs, but the selection in vitro of variants in which the MICs were increased 4–16‐fold after 10 serial passages in subinhibitory concentrations of enrofloxacin suggests that inappropriate use might favour the development of resistant strains in vivo.     

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.     

Antimicrobial susceptibilities and bacteriological characteristics of bovine Pseudomonas aeruginosa and Serratia marcescens isolates from mastitis

The presence of metallo-β-lactamase (MBL)-producing and multidrug-resistant Pseudomonas aeruginosa (MDRP) strains among bovine isolates of Gram-negative bacilli, and O-serotypes of bovine Serratia marcescens and P. aeruginosa isolates have been reported rarely. The aims of this study were to (1) elucidate antimicrobial susceptibilities and O-serotypes of P. aeruginosa and S. marcescens isolates from bovine mastitis and the presence of MBL-producers and MDRP strains among them and (2) evaluate their relationships to human isolates. We investigated the MICs of 24 antimicrobials and O-serotypes for 116 P. aeruginosa and 55 S. marcescens isolates in Japan, primarily in 2006. A total of 171 isolates exhibited high antimicrobial susceptibilities with the exception of a partial drug. P. aeruginosa isolates exhibited high susceptibilities of ≥ 95.7% to ciprofloxacin, imipenem, meropenem, piperacillin, ceftazidime, cefepime, cefoperazone/sulbactam, amikacin, tobramycin, and gentamicin; however, they exhibited a susceptibility of only 69.8% to aztreonam. They exhibited substantial resistances to ceftriaxone, enrofloxacin, cefotaxime, and moxalactam. S. marcescens isolates exhibited high susceptibilities of ≥ 90.9% to kanamycin, ceftiofur, sulfamethoxazole-trimethoprim, and the 15 aforementioned drugs, but exhibited resistance to minocycline. Neither MBL-producers nor MDRP strains were detected among the 171 strains. The dominant serotypes of P. aeruginosa isolates were OG, OA, OB, OI, OF, OE, and OK; those of S. marcescens isolates were O6 and O5. Every S. marcescens isolate was pigmented. These findings suggest that bovine P. aeruginosa and S. marcescens isolates differ from human isolates from both antibiogram and phenotypic perspectives, and could help to evaluate differences in bacteriological characteristics between bovine and human isolates.     

Antimicrobial resistance of Providencia spp. isolated from animal manure

Providencia spp. are Gram negative bacteria, belonging to the family Enterobacteriaceae and are responsible for gastrointestinal disorders both in humans and animals. We report on the isolation of multi-drug resistant Providencia spp. (n=4) from animal manure. The isolates were found to be resistant to clindamycin and to tetracycline, as well as to the macrolide and sulphonamide groups of antibiotics.     

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.     

Antimicrobial resistance in streptococcal species isolated from bovine mammary glands

Streptococcal species isolated from dairy cows with clinical mastitis were obtained from mastitis research workers in Florida, Louisiana, New York, Vermont, Washington, and West Virginia. Seventy-one streptococcal isolates were tested, including 39 strains of Streptococcus agalactiae, 21 strains of S dysgalactiae, and 11 strains of S uberis. The minimal inhibitory concentration of erythromycin, lincomycin, oxytetracycline, penicillin, spectinomycin, streptomycin, and tetracycline was determined for each isolate. Differences were not detected among strains with respect to geographic origin. None of the strains was resistant to penicillin. Lincomycin was the next most effective antimicrobial, with only 2 resistant strains of each streptococcal species. There were no differences among the streptococcal species with respect to resistance to either penicillin or lincomycin. Streptococcus uberis was more likely to be resistant to erythromycin than were S agalactiae and S dysgalactiae (P less than 0.02). Streptococcus agalactiae and S uberis had similar distributions for resistance to oxytetracycline, tetracycline, spectinomycin, and streptomycin. Strains of S dysgalactiae were more likely to have intermediate resistance to oxytetracycline and streptomycin than were strains of S agalactiae and S uberis, which were highly resistant to oxytetracycline and streptomycin (P less than 0.001). Differences were not detected among the streptococcal species with respect to resistance to spectinomycin. Resistance to multiple antimicrobials was observed in all streptococcal species tested. Although S dysgalactiae appeared to have a greater percentage of strains (73%) that were resistant to multiple antimicrobials than did S agalactiae (31%) or S uberis (45%), differences were not statistically significant.     

Monosaccharide inhibition of adherence by Pseudomonas aeruginosa to canine corneocytes

The effect of d -galactose, d -mannose, l -rhamnose and dextrose on the adhesion to canine corneocytes by three strains of Pseudomonas aeruginosa was studied in six healthy dogs. Canine corneocytes were collected from the inner aspect of the pinna using adhesive discs (D-Squame®). Half millimetre of bacterial suspension in phosphate-buffered saline (PBS) with or without the addition of a monosaccharide was placed over the corneocyte layer and incubated in moist chambers. Image analysis was used to quantify bacterial adherence to corneocytes. The three strains of Pseudomonas adhered well to canine corneocytes. All monosaccharides tested inhibited the adherence of Pseudomonas to canine corneocytes. The mean reduction in adhesion for individual sugars at a concentration of 0.1% was 40.2% (dextrose), 30.8% ( l -rhamnose), 25.6% ( d -galactose) and 19.4% ( d -mannose). When d -galactose, d -mannose and l -rhamnose were used in combination at 0.1% concentration, the mean reduction in adherence was 52.9%. The monosaccharides studied may have a potential role in the management of Pseudomonas infections in dogs.     

水貂铜绿假单胞菌鞭毛分型及分离株flic基因的克隆与序列分析

为分析铜绿假单胞菌(PA)分离株鞭毛蛋白基因(flic)之间的差异,本研究通过PCR方法扩增由山东地区发病水貂的肺脏组织中分离的29株分离株flic基因的部分片段,根据其片段长度确定PA的鞭毛类型。选取两种类型代表株(PASD01、PASD03),采用PCR方法扩增flic全基因进行序列分析。结果表明,PASD01分离株与国外A型PA flic基因序列同源性为99.6%～99.7%,PASD03分离株与B型PA flic基因的序列同源性为98.9%～99.5%,而两个分离株之间的序列同源性为76%。本研究首次从水貂病变组织中克隆得到两种鞭毛蛋白类型的flic基因,该基因在各型鞭毛菌株中高度保守,为进一步研制鞭毛亚单位疫苗提供实验依据。     

Antimicrobial resistance in enteric pathogens isolated from Minnesota pigs from 1995 to 2004

This study investigated the occurrence and antimicrobial resistance profiles of Escherichia coli and Salmonella sp. isolated from swine samples submitted to the Minnesota Veterinary Diagnostic Laboratory (MVDL) in Saint Paul, Minnesota from 1995 to 2004. During this time period, a total of 5072 E. coli and 2793 Salmonella sp. was isolated. Most of these isolates were found to be resistant to the tetracycline and beta-lactam group of antibiotics. Resistance to spectinomycin was also frequently observed. An increasing trend in ampicillin resistance and a decreasing trend in apramycin resistance were seen in both pathogens, although ampicillin resistance was relatively higher in E. coli than in Salmonella. Aminoglycoside (amikacin) and quinolone (enrofloxacin) were the only antimicrobials to which minimum or no resistance was observed. The resistance of pig pathogens to several antibiotics indicates the need to routinely monitor the use of these antimicrobials and their associated resistance in pig populations.     

Antimicrobial resistance of thermophilic Campylobacter spp. isolated from cattle in Poland

Campylobacter species are among the most frequently identified bacterial causes of human gastroenteritis. Because Campylobacter spp. harbored by cattle can be transmitted to humans, in this study we investigated antimicrobial resistance of thermophilic Campylobacter isolated from cows. Our study included 150 strains of Campylobacter (143 strains of C. jejuni and 7 strains of C. coli) isolated from cows in South-Western Poland. The minimal inhibitory concentration (MIC) to ciprofloxacin, erythromycin, gentamicin and tetracycline were determined using the agar dilution methodology. All strains of C. coli were susceptible to all four drugs studied. The most frequently detected resistance of C. jejuni was to ciprofloxacin (26 strains 18.2%). Resistance to tetracycline was observed in 5 strains (3.5%). All strains of C. jejuni were susceptible to erythromycin and gentamicin.     

Phenotypic and genetic characterization of Edwardsiella tarda isolated from pond sediments

The extent of genotypic and phenotypic diversity of Edwardsiella tarda isolated from pond sediment was assessed by SDS-PAGE, Plasmid Profiling and ERIC-PCR. SDS-PAGE of whole cell protein extracts reveals 20-23 discrete bands with molecular wt of 14-110 kDa. Several bands with molecular weight range of 38-83 kDa were present in all the isolates. Numerical analysis of protein electrophoregram delineated the isolates into four clusters. Two different types of plasmids having molecular mass of 23 kDa and 29 kDa were obtained by plasmid profiling. About 51% of the isolates carried both the plasmids. ERIC-PCR generates 3-7 bands with molecular mass of 14-1013 bp. Numerical analysis differentiated the ERIC pattern into 5 clusters at 60% similarly level. It was concluded that out of three methods ERIC-PCR was found to be more sensitive for intraspecific typing of E. tarda and can be used as a potential tool for epidemiological studies in the future.     

犬细小病毒山东分离株VP2基因的克隆与遗传变异分析

为了解山东省犬细小病毒(CPV)流行毒株的遗传变异情况及其与犬细小病毒病的分子流行病学关系,对2013年分离到的12株山东CPV流行毒株进行了VP2基因克隆测序与同源性分析,绘制系统进化树。结果表明,获得的VP2基因的全长为1 755 bp,编码584个氨基酸残基;各分离毒株之间的核苷酸和氨基酸同源率分别为98.9%~100.0%和98.8%~100.0%,与Gen Bank公布的国内外参考毒株核苷酸和氨基酸同源率在98%以上,分离株VP2基因及其推导的氨基酸的变异主要以点突变为主,关键氨基酸位点未见明显改变。遗传进化分析显示,有8株为New CPV-2a,4株为New CPV-2b,表明目前山东省所流行的CPV主要以New CPV-2a亚型为主。研究结果为山东犬细小病毒病的防控提供了依据。