Rapid isolation of Brachyspira hyodysenteriae and Brachyspira pilosicoli from pigs

The aim of this study was to compare and evaluate the time required to isolate Brachyspira hyodysenteriae and Brachyspira pilosicoli from porcine faeces. This was done using previously described selective media (spectinomycin) S400, (colistin, vancomycin and spectinomycin) CVS and (spectinomycin, vancomycin, colistin, spiramycin and rifampin with swine faecal extract) BJ, compared with the method based on blood agar modified medium, with spectinomycin and rifampin (BAM-SR), including a pre-treatment step. Fourteen spirochaetal strains were obtained in pure cultures after 5 days (48 h in BAM-SR primary plate and three passages every 24 h in brain heart infusion (BHI) without antibiotics) pre-treating simulated samples in brain heart infusion broth with spectinomycin (400 microg/ml) and rifampin (15 microg/ml), before streaking on the selective BAM-SR medium. Spirochaetes from samples in S400, CVS and BJ, with and without pre-treatment, were obtained in pure cultures only after repeatedly transferring on plates of the same selective medium requiring 15-18 days according to the strain. BAM-SR used after the pre-treatment step showed a detection limit ranging from 3.5 x 10(2) to 6.7 x 10(7) cells/g faeces and was the only method able to support the growth of spirochaetes after 48 h.     

Effect of antibiotics and vehicles on bovine mammary polymorphonuclear leukocyte morphologic features, viability, and phagocytic activity in vitro

Effects of antibiotics and antibiotic vehicles on polymorphonuclear leukocytes (PMNL) isolated from bovine mammary glands were studied in vitro. Amikacin, dicloxacillin, erythromycin, gentamicin, lincomycin, nitrofurantoin, novobiocin-penicillin, polymyxin B, rifampin, tetracycline, or tiamulin was added to culture medium at 1 mg/ml and chloramphenicol was added at 4 mg/ml. Drug concentrations were equivalent to those detected in milk immediately after injection into the mammary gland. Vehicles included mineral oil and peanut oil, each at a dilution of 1:100 in culture medium. The PMNL morphologic features, viability, and phagocytic activity were evaluated. In comparison with the phosphate-buffered saline solution (PBSS) control, significant (P less than 0.05) alterations in normal cell morphologic features were observed in PMNL cultured with tetracycline, nitrofurantoin, erythromycin, polymyxin B, rifampin, novobiocin-penicillin, chloramphenicol, tiamulin, or peanut oil. Viabilities of PMNL cultured with chloramphenicol, novobiocin-penicillin, or tiamulin were significantly (P less than 0.05) reduced when compared with those of PBSS controls. Addition of Staphylococcus aureus to culture medium enhanced morphologic alterations and reduced viabilities of PMNL. Phagocytosis of S aureus by PMNL was significantly (P less than 0.05) depressed in medium containing novobiocin-penicillin, amikacin, rifampin, chloramphenicol, tiamulin, or peanut oil in comparison with that of PMNL incubated in PBSS.     

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.     

Pharmacokinetics of single intravenous and single and multiple dose oral administration of rifampin in mares

The disposition of rifampin in six healthy mares after single intravenous (i.v.) and oral (p.o.) doses and after seven oral doses of 10 mg/kg administered twice a day was investigated using a high performance liquid chromatographic (HPLC) method. Pharmacokinetic variables for rifampin determined using the HPLC method were comparable to variables reported from earlier studies utilizing a microbiological assay. Desascetylrifampin, a major metabolite of the parent compound, could not be detected in the serum but was detected at low concentrations in urine. Mean trough concentrations of rifampin increased from the first to the second dose of the multiple dose oral study and then remained unchanged through 72 h. At 84 h after the first dose (i.e. 12 h after the final dose) the rifampin concentration was significantly decreased ( P = 0.001). The harmonic mean of the half-life of rifampin decreased significantly from 13.3 h after a single oral dose of 7.99 h after the seventh oral dose. The mean serum protein binding of rifampin over the concentration range of 2–20 μg/ml was 78%. Mean trough serum concentrations of unbound rifampin after multiple oral doses ranged from 0.67 μg/ml at 24 h to 0.40 μg/ml at 72 h. The mean unbound serum rifampin concentration at 84 h (i.e., 12 h after the final dose) was 0.30 μg/ml. Trough concentrations and the 84-h sample concentration of unbound rifampin exceeded the minimum inhibitory concentration for most gram positive bacterial isolates from horses reported in this study. All organisms with minimum inhibitory concentrations less than 0.125 μg/ml were considered susceptible.
Based on the pharmacokinetics of rifampin after p.o. administration, we concur with the current dosage recommendation of 10 mg/kg twice a day by mouth. At this dose, most streptococci, Rhodococcus equi , and coagulase-positive staphylococci would be considered susceptible to rifampin.     

Comparison of partial ribosomal DNA sequences of Babesia gibsoni occurring in Miyazaki prefecture, Japan.

Nucleotide sequences of ribosomal DNA (rDNA) of Babesia (B.) gibsoni occurring in Miyazaki, western Japan, were examined using blood samples obtained from seven dogs suffering from natural canine babesiosis. DNA isolated from these blood samples was subjected to the polymerase chain reaction (PCR). The nucleotide sequences of the PCR products were determined and compared with other rDNA sequences of B. gibsoni isolated from Asia, Europe and U.S.A. Although homology values between our isolates and those isolated from Europe and U.S.A. were both 84.0%, respectively, our isolates were identical to the Asian types. In conclusion, B. gibsoni occurring in Miyazaki was revealed to have the genotype Asia 1 or Asia 2 from a comparison of the partial rDNA sequences.     

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.     

Characterization of antimicrobial resistance of Salmonella Newport isolated from animals, the environment, and animal food products in Canada

Multi-drug-resistant (MDR) Salmonella enterica serovar Newport strains are increasingly isolated from animals and food products of animal origin and have caused septicemic illness in animals and humans. The purpose of this study was to determine the occurrence and the epidemiologic, phenotypic, and genotypic characteristics of S. Newport of animal origin that may infect humans, either via the food chain or directly. During the 1993-2002 period, the Office International des Epizooties Reference Laboratory for Salmonellosis in Guelph, Ontario, received 36 841 Salmonella strains for serotyping that had been isolated from animals, environmental sources, and food of animal origin in Canada. Of these, 119 (0.3%) were S. Newport. Before 2000, none of 49 S. Newport strains was resistant to more than 3 antimicrobials. In contrast, between January 2000 and December 2002, 35 of 70 isolates, primarily of bovine origin, were resistant to at least 11 antimicrobials, including the extended-spectrum cephalosporins. The blaCMY-2', flo(st'), strA, strB, sulII, and tetA resistance genes were located on plasmids of 80 to 90 MDa that were self-transmissible in 25% of the strains. Conserved segments of the integron 1 gene were found on the large MDR-encoding plasmids in 3 of 35 strains additionally resistant to gentamicin and spectinomycin or to spectinomycin, sulfamethoxazole-trimethoprim, and trimethoprim. Resistance to kanamycin and neomycin was encoded by the aphA-1 gene, located on small plasmids (2.3 to 6 MDa). The increase in bovine-associated MDR S. Newport infections is cause for concern since it indicates an increased risk of human acquisition of the infection via the food chain.     

鸡源沙门菌的耐药性及脉冲场凝胶电泳分型研究

为了解北京地区鸡源沙门菌血清型的分布、耐药性以及基因型情况,利用Riboprinter全自动微生物基因指纹鉴定仪对分离到的沙门菌进行血清型鉴定,采用微量肉汤稀释法进行药物敏感性测定,应用脉冲场凝胶电泳(PFGE)进行基因分型.结果显示,共分离到15株肠炎沙门菌、1株鼠伤寒沙门菌,沙门菌对利福平、萘啶酸和多黏菌素E的耐药...     

Epidemiology of methicillin-resistant Staphylococcus aureus in dairy herds.

Sixty-eight methicillin-resistant Staphylococcus aureus strains were isolated from mastitis milk samples originating from 20 Belgian dairyherds. All these strains appeared to be representatives of one single strain which was probably of human origin. Evidence is presented indicating a rapid in vivo evolutionary change in this strain. The following characteristics were found to be variable: the production of beta haemolysin inversely connected with fibrinolysin (staphylokinase) activity; the production of lipase, enterotoxin B and delta haemolysin; the resistance to neomycin, chloramphenicol, tetracyclines, methicillin and spectinomycin associated with constitutive or inducible macrolide resistance.     

Antimicrobial sensitivity testing of Australian isolates of Bordetella avium and the Bordetella avium-like organism

SUMMARY: The in-vitro sensitivity of 16 Australian isolates of Bordetella avium and 15 isolates of B avium -like organism to 11 antimicrobial agents or combinations of agents was determined using a microtitre plate system to establish minimal inhibitory concentrations. All the B avium isolates were sensitive to ampicillin but resistant to erythromycin, lincomycin, spectinomycin, sulphamethoxazole, trimethoprim, and lincomycin + spectinomycin. Most of the B avium isolates were sensitive to tetracycline and resistant to streptomycin and sulphadiazine. All the B avium -like isolates were resistant to ampicillin, erythromycin, lincomycin, spectinomycin, streptomycin, tetracycline, trimethoprim, and lincomycin + spectinomycin. Most B avium -like isolates were sensitive to sulphadiazine, sulphamethoxazole and trimethoprim-sulphamethoxazole.     

乌鳢舒氏气单胞菌的分离鉴定及药敏试验

对广东省佛山市某乌鳢养殖鱼场患病的乌鳢进行病原分离,将疑似分离菌S-1株进行形态和培养特性、生化特性鉴定、PCR鉴定、药敏试验和人工感染等试验。结果表明,该分离菌为舒氏气单胞菌。药敏试验结果表明,本分离菌对壮观霉素、米诺环素、庆大霉素、氧氟沙星、诺氟沙星、头孢类等多种药物敏感。本研究为舒氏气单胞菌病细菌学检测及治疗提供了科学依据。     

Streptococcus agalactiae and Streptococcus difficile 16S-23S intergenic rDNA: genetic homogeneity and species-specific PCR

Streptococcus difficile is a non-hemolytic Gram-positive bacterial coccus that causes septicemia and meningoencephalitis in farmed tilapia (Oreochromis sp.) and rainbow trout (Oncorhynchus mykiss). Recent studies have demonstrated S. difficile to be a group B, type Ib streptococcus with a whole cell protein electrophoretic profile indistinguishable from S. agalactiae and a biochemical profile similar to that observed for other group B, type Ib streptococci isolated from fish and frogs. The aim of this study was to expand on these findings by comparative nucleic acid sequence analysis of the 16S-23S ribosomal DNA (rDNA) intergenic spacers of S. difficile and S. agalactiae. The 97.7% sequence homology identified in these studies supports the taxonomic relationship of these two organisms. The sequence data generated were also used to construct a pair of species-specific PCR primers for use in molecular detection and identification schemes.     

Isolation,Identification and Drug Susceptibility Test of Pseudomonas aeruginosa in Chickens

In order to detect the main bacterial pathogens and analyze the drug resistance for providing therapeutical guidance for drug administration,4 stains were isolated from death chickens of a breeding chicken farm in Qinhuangdao area.All 4 isolates were identified by morphological characteristics,biochemical test,immunofluorescent staining,amplification of 16S rRNA genes by PCR,sequence analysis and drug susceptibility test.The results showed that all 4 isolates were gram-negative,bacillus brevis with green fluorescent,and the sequence of 16S rRNA gene of isolates were analyzed and test results showed that specific amplified band was consistent with the expected size.The isolated bacteria were identified to be Pseudomonas aeruginosa.Results of drug susceptibility test indicated that the bacterial isolates were highly sensitive to gentamycin,amikacin,spectinomycin,ceftazidine,ceftriaxone,levofloxacin,enrofloxacin,florfenicol,polymyxin B and fosfomycin,intermidiate sensitive to tetracycline and doxycyclin,resistant to neomycin,cefotaxime,cephalothin,amoxicillin,ampicillin,erythromycin,trimethoprim-sulfamethoxazole and oxygen sulfamethoxazole isoxazole.The results suggested that the resistances of Pseudomonas aeruginosa isolates to commonly used antibiotics were serious.So antibiotics should be reasonably selected based on the result of drug susceptibility test in this farm.     

Drug resistance and biochemical characteristics of Salmonella from turkeys.

A study was conducted to determine the antibiotic resistance and biochemical characteristics of 2690 Salmonella strains belonging to 52 serovars and isolated from environmental and feed samples from 270 turkey flocks in Canada. Resistance of the Salmonella strains to the aminoglycoside antibiotics varied widely; none of the strains were resistant to amikacin, 14.2% were resistant to neomycin, 25.8% were resistant to gentamicin, and 27.7% of the strains were resistant to kanamycin. Most strains (97.6%) were resistant to the aminocyclitol, spectinomycin. Regarding resistance to the beta-lactam antibiotics, 14.3% and 14.4% of the strains were resistant to ampicillin and carbenicillin, respectively, whereas only 5 (0.2%) of the strains were resistant to cephalothin. None of the strains were resistant to the fluoroquinolone ciprofloxacin or to polymyxin B. Resistance to chloramphenicol and nitrofurantoin was found in 2.4% and 7% of the strains, respectively. Only 1.7% of the strains were resistant to the trimethoprimsulfamethoxazole combination, whereas 58.1% were resistant to sulfisoxazole. Thirty-eight percent of the strains were resistant to tetracycline. Salmonella serovars differed markedly in their drug resistance profiles. Biochemical characterization of the Salmonella showed that the S. anatum, S. saintpaul and S. reading serovars could be divided into distinct biotypes.     

抗菌药物联合使用对耐达氟沙星鸡毒支原体的体外抑制作用研究

为探究抗菌药物联合使用对耐达氟沙星鸡毒支原体的体外抑制效果,选择鸡毒支原体标准株S6和对达氟沙星MIC升高程度不同的突变株M1、M2和M4,分别测定泰乐菌素、大观霉素、林可霉素、泰妙菌素、替米考星和多西环素种对4株鸡毒支原体的最低抑菌浓度,并使用达氟沙星分别联合上述6种抗菌药物进行体外抑菌试验.结果显示:除达氟沙星外,...     

鸡源绿脓杆菌的分离鉴定及药敏试验

为确诊引起秦皇岛某鸡场细菌性疾病的主要病原菌,分析其耐药情况,为临床用药和治疗提供依据,本试验对送检的病死鸡肝脏中分离得到的4株菌进行形态特征观察、生化鉴定、免疫荧光染色观察、细菌16S rRNA基因PCR扩增与测序分析及药敏试验.结果显示,4株分离菌均为革兰氏阴性、带绿色荧光的短杆菌,PCR扩增出的特异性条带与预期大小相吻合,确定为绿脓杆菌.药敏试验结果显示,分离株对庆大霉素、阿米卡星、大观霉素、头孢他啶、头孢曲松、左氧氟沙星、恩诺沙星、氟苯尼考、多黏菌素B和磷霉素高度敏感,对四环素和强力霉素中度敏感,而对新霉素、头孢噻肟、头孢噻吩、阿莫西林、氨苄西林、红霉素、复方新诺明和磺胺甲氧异唑耐药.提示,该分离菌对常用抗生素耐药情况严重,养殖场应根据药敏试验结果选择敏感药物进行合理用药.     

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.     

Multidrug resistance and distribution of Salmonella serovars in slaughtered pigs

The present study was undertaken to estimate the occurrence and distribution of multidrug resistance (MDR) among Salmonella serovars isolated from slaughtered pigs at Debre Zeit, Ethiopia. A total of 501 different samples were examined of which 42 (41.6%) of 101 mesenteric lymph nodes, 22 (21.8%) of 101 tongues, 17 (16.8%) of 101 caecal contents, 11 (11.1%) of 99 livers and two (2%) of 99 muscle (diaphragm and abdomen) samples were Salmonella positive. Of the 94 Salmonella isolates representing 15 different serovars, 69 (73.4%) were multidrug resistant (resistance to two or more antimicrobials). Among the Salmonella serovars a high level of MDR was observed in S. Hadar, S. Kentucky, S. Blockley and S. Enteritidis mainly to tetracycline (88.6%), streptomycin (82.9%), nitrofurantoin (74.3%), nalidixic acid and ciprofloxacin (42.9% each), sulfisoxazole (21.1%) and spectinomycin (20%). The pattern of MDR varied from two to eight antimicrobials among the resistant Salmonella serovars. The common profiles of resistance among the MDR serovars were the combined resistance to nitrofurantoin, streptomycin and tetracycline (R type NitStrTet, 51.4%), ciprofloxacin, nalidixic acid and nitrofurantoin (R type CipNalNit, 10%), ciprofloxacin, nalidixic acid, spectinomycin, streptomycin, sulfisoxazole and tetracycline (R type CipNalSptStrSulTet, 14.3%) and to ciprofloxacin, kanamycin, nalidixic acid, neomycin, nitrofurantoin, streptomycin and tetracycline (R type CipKanNalNeoNitStrTet, 10%). Results of the present study indicate the widespread occurrence and distribution of MDR Salmonella serovars in slaughtered pigs which could be a potential source of human MDR Salmonella infections.     

一起犊牛冠状病毒、轮状病毒与大肠埃希氏菌混合感染的实验室诊断

陕西省某规模化奶牛场发生一起新生犊牛腹泻病例,为了确定发病原因,通过临床剖检,无菌采集病料进行细菌分离鉴定、PCR检测病毒等技术,最终确诊这起疫病为牛冠状病毒(BCoV)、牛轮状病毒(BRV)和大肠埃希氏菌混合感染所致的犊牛腹泻。对分离到的大肠埃希氏菌进行药敏试验,结果显示分离菌对左氟沙星和诺氟沙星敏感,对四环素、复方新诺明、多黏菌素B、卡那霉素、庆大霉素、多西环素、氨苄西林、大观霉素均耐药。     

Antimicrobial susceptibility of clinical strains of Streptococcus suis isolated from pigs in Spain

The antimicrobial susceptibility of 151 clinical Streptococcus suis strains isolated from diseased pigs in Spain was determined by a microdilution method. Isolates were mostly susceptible to beta-lactam antimicrobials, aminoglycosides, enrofloxacin, novobiocin and spectinomycin. More than 87% of the S. suis isolates were resistant to tetracyclines, sulphonamides, macrolides and clindamycin. Strains of serotype 9 were significantly more resistant than strains of serotype 2 (P<0.05) to tylosin (94% versus 77%) and clindamycin (94% versus 64%). Eighty-seven percent of the S. suis isolates were resistant to at least four antimicrobials and nine isolates (6%) were resistant to at least six antimicrobials. The most frequently identified multidrug pattern involved resistance against tetracyclines, sulphonamides, macrolides and lincosamides, with 69% of the isolates exhibiting this resistotype. Fifteen out of the 22 strains of serotype 2 (68.2%), and 84 out of the 98 of the strains of serotype 9 (85.7%) exhibited this resistotype, indicating its widespread distribution among the strains of the two most frequently isolated serotypes.