Presence of arsenic resistance in Salmonella enterica serovar Kentucky and other serovars isolated from poultry

A collection of 125 Salmonella enterica poultry isolates (71 serovar Kentucky isolates, and the remainder belonging to serovars Alachua, Enteritidis, Hadar, Heidelberg, Montevideo, Mbandaka, Senftenberg, Typhimurium, and Worthington) were tested for the ability to grow on tryptic soy agar containing sodium arsenite [As(III)] or arsenate [As(V)]. All serovar Kentucky isolates and 18 of the non-Kentucky isolates were able to grow in the presence of 0.1 mM As(III), and 69 grew in the presence of 1 mM As(V). Thirty of the non-Kentucky isolates did not grow at these As(III) and As(V) concentrations, but seven grew at 1 mM As(III) and 10 mM As(V). PCR-based analysis demonstrated the presence of arsB and arsD sequences in all Kentucky isolates, whereas one or both of these sequences were present in only 30 of the other isolates. It remains to be determined if these arsenic-resistance determinants benefit Salmonella exposed to man-made arsenic-containing compounds in poultry environments.     

Prevalence of Haemophilus parasuis serovars isolated in Spain from 1993 to 1997

From 1993 to 1997, 327 strains of Haemophilus parasuis were isolated from spanish swine in our Diagnostic Laboratory and 174 strains (53.2%) were serotyped. Four serotypes, sv. 5 (18.4%), sv 4 (16%), sv. 2 (9.2%) and sv. 13 (8%) were the most frequently isolated and 29.3% of the studied strains were classified as non typable. The results obtained indicate that the distribution of the serotypes in Spain is very similar to that found by other researchers in Germany, Australia, Canada and alike to that found in the United States.     

Antimicrobial resistance in Salmonella enterica serovars Enteritidis and Typhimurium isolated from animals in Korea: comparison of phenotypic and genotypic resistance characterization

Fourteen and 22 each of Salmonella Enteritidis and Salmonella Typhimurium (S. Typhimurium) were isolated from animals from 1983 to 1999 in Korea and tested for their antibiotic resistance patterns, phage types and resistance gene patterns. S. Typhimurium isolates were highly resistant to streptomycin, sulfisoxazole and tetracycline, 95, 95 and 86%, respectively. The incidence of multiple antibiotic resistance (resistant to more than two drugs tested) of S. Typhimurium isolates was extremely high (100%) comparing to S. Enteritidis isolates (21%). Two of the five ACSSuT (ampicillin, chloramphenicol, streptomycin, sulfisoxazole and tetracycline) resistant type S. Typhimurium isolates were phage type definitive type 104 (DT104).For the detection of resistance related genes in S. Enteritidis and S. Typhimurium isolates, particularly ACSSuT type S. Typhimurium, antibiotic resistance genes, cmlA/tetR, bla(PSE-1) and bla(TEM), and genus Salmonella specific gene, sipB/C, were amplified using four pairs of primers in a hot-start multiplex polymerase chain reaction (PCR). Two Korean isolates of S. Typhimurium DT104 showed bla(TEM) amplicons instead of bla(PSE-1) for the ampicillin resistance and they were susceptible to florfenicol. The multiplex PCR used in this study was useful in characterization of multiple drug resistant Salmonella isolates, especially ACSSuT type S. Typhimurium, and identification of beta-lactamase gene distribution among Salmonella 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.     

High prevalence of antibiotic resistance in pathogenic Escherichia coli from large- and small-scale poultry farms in Bangladesh

Antibiotic resistance in avian bacterial pathogens is a common problem in the Bangladesh poultry industry. The aim of the present study was to provide information on the present status of antibiotic resistance patterns in avian pathogenic Escherichia coli in Bangladesh. Of 279 dead or sick poultry of different ages, 101 pathogenic E coli strains isolated from broilers and layer hens with colibacillosis infections were screened to determine phenotypic expression of antimicrobial resistance against 13 antibiotics used in both veterinary and human medicine in Bangladesh. Of 101 pathogenic E. coli isolates, more than 55% were resistant to at least one or more of the tested compounds, and 36.6% of the isolates showed multiple-drug-resistant phenotypes. The most common resistances observed were against tetracycline (45.5%), trimethoprim-sulphamethoxazole (26.7%), nalidixic acid (25.7%), ampicillin (25.7%), and streptomycin (20.8%). Resistance to ciprofloxacin (12.9%), chlormaphenicol (8.9%), nitrofurantoin (2%), and gentamicin (2%) was also observed, and none of the isolates were resistant to tigecycline as well as extended spectrum beta-lactamase (ESBL) producers. One isolate was resistant to cefuroxime (1%), cefadroxil (1%), and mecillinam (1%) but was not an ESBL producer. Resistance rates, although significant in Bangladeshi isolates, were found to be lower than those reported for avian isolates from the Republic of Korea and clinical, avian, and environmental isolates from Bangladesh. The high level of antibiotic resistance in avian pathogens from Bangladesh is worrisome and indicates that widespread use of antibiotics as feed additives for growth promotion and disease prevention could have negative implications for human and animal health and the environment.     

A temporal study of Salmonella serovars from environmental samples from poultry breeder flocks in Ontario between 1998 and 2008

A temporal study was carried out to determine Salmonella prevalence, trends, major serovars, and their clusters from environmental samples, in poultry breeder flocks in Ontario between January 1998 and December 2008. Surveillance data were obtained from the Ontario Hatchery and Supply Flock Policy. Logistic regression with a random effect for flock was used to identify factors [poultry type, year (trend) and season] associated with the prevalence of Salmonella. A cluster detection test was used to identify clusters of common serovars. The period prevalence of Salmonella was 47.4% in broiler-breeder, 25.7% in layer-breeder, and 19.6% in turkey-breeder flocks. The overall trend in the prevalence of Salmonella was decreasing for all breeder types, due primarily to decreasing trends of Salmonella Heidelberg. The seasonal effects varied by year with the highest probability of Salmonella occurring in different seasons. The 4 most common serovars identified were Salmonella Heidelberg, Kentucky, Hadar, and Typhimurium in broiler-breeders; Salmonella Heidelberg, Brandenburg, Thompson, and Typhimurium in layer-breeders; and Salmonella Heidelberg, Saintpaul, Brandenburg, and Muenster in turkey-breeders. Salmonella Enteritidis was infrequently isolated in all poultry breeder types. Temporal clusters of different serovars were identified in all poultry breeder types. Clusters of Salmonella Heidelberg, Typhimurium, and Hadar from environmental samples from breeder flocks were detected during a similar period to clusters from hatchery fluff samples from the same population. Therefore, interventions at the breeder flock-level might help to reduce transmission of Salmonella from breeder flocks to hatcheries and possibly, to lower levels of the poultry production chain.     

Comparative investigation of prevalence and antimicrobial resistance of Salmonella between pasture and conventionally reared poultry

The objective of this study was to compare Salmonella prevalence and antimicrobial resistance between pasture and conventional poultry farms. We collected the first Salmonella prevalence and antimicrobial resistance data on pasture poultry farms. Fecal droppings were collected from 31 farms from Wisconsin (nine farms from each production type) and the Southeast (North Carolina, Virginia, and South Carolina; five conventional and 10 pasture poultry farms) in a 1-yr period. The specimens were cultured for Salmonella and tested for resistance to 12 antimicrobials. A univariate analysis was conducted to determine the significant differences in prevalence and resistance. At the farm level, no significant difference in Salmonella prevalence was found on 33% pasture and 47% conventional poultry farms (P= 0.4928). On an individual specimen level, flocks reared conventionally had higher prevalence than in pasture (P < 0.0001). Of all the isolates found to have resistance, 80% were from the Southeast. Of all the pasture isolates tested, 5% (8 of 162) were resistant to ceftriaxone, which is higher than previously reported from any production setting. None of the isolates from conventional flocks showed resistance to ceftriaxone. Multidrug resistance (resistance to three or more classes of antimicrobials) was found in 69% of the isolates from conventional farms and 11% on pasture farms in the Southeast (P < 0.0001), with the predominant resistance type of AmCSSuTeAx (ampicillin, chloramphenicol, streptomycin, sulfasoxazole, tetracycline, amoxicillin/clavulanic acid; 62%). About 5% of the pasture isolates from the Southeast showed the AmAxCFCRO (ampicillin, amoxicillin/clavulanic acid, cephalothin, ceftriaxone) multidrug resistance pattern. None of the isolates from Wisconsin were found to be multidrug resistant.     

Profile of selected bacterial counts and Salmonella prevalence on raw poultry in a poultry slaughter establishment.

The USDA Food Safety and Inspection Service determined populations of bacteria on poultry during processing at a slaughter plant in Puerto Rico in November and December 1987. The plant was selected because of its management's willingness to support important changes in equipment and processing procedures. The plant was representative of modern slaughter facilities. Eight-hundred samples were collected over 20 consecutive 8-hour days of operation from 5 sites in the processing plant. Results indicated that slaughter, dressing, and chilling practices significantly decreased the bacterial contamination on poultry carcasses, as determined by counts of aerobic bacteria, Enterobacteriaceae, and Escherichia coli. Salmonella was not enumerated; rather, it was determined to be present or absent by culturing almost the entire rinse. The prevalence of Salmonella in the study decreased during evisceration, then increased during immersion chilling.     

Characterization of in vivo-acquired resistance to macrolides of Mycoplasma gallisepticum strains isolated from poultry

ABSTRACT: The macrolide class of antibiotics, including tylosin and tilmicosin, is widely used in the veterinary field for prophylaxis and treatment of mycoplasmosis. In vitro susceptibility testing of 50 strains of M. gallisepticum isolated in Israel during the period 1997-2010 revealed that acquired resistance to tylosin as well as to tilmicosin was present in 50% of them. Moreover, 72% (13/18) of the strains isolated from clinical samples since 2006 showed acquired resistance to enrofloxacin, tylosin and tilmicosin. Molecular typing of the field isolates, performed by gene-target sequencing (GTS), detected 13 molecular types (I-XIII). Type II was the predominant type prior to 2006 whereas type X, first detected in 2008, is currently prevalent. All ten type X strains were resistant to both fluoroquinolones and macrolides, suggesting selective pressure leading to clonal dissemination of resistance. However, this was not a unique event since resistant strains with other GTS molecular types were also found. Concurrently, the molecular basis for macrolide resistance in M. gallisepticum was identified. Our results revealed a clear-cut correlation between single point mutations A2058G or A2059G in domain V of the gene encoding 23S rRNA (rrnA, MGA_01) and acquired macrolide resistance in M. gallisepticum. Indeed, all isolates with MIC ≥ 0.63 μg/mL to tylosin and with MIC ≥ 1.25 μg/mL to tilmicosin possess one of these mutations, suggesting an essential role in decreased susceptibility of M. gallisepticum to 16-membered macrolides.     

禽源金黄色葡萄球菌耐药性监测

从临床样品中分离鉴定得到183株金黄色葡萄球菌,采用微量肉汤稀释法监测其对24种抗菌药物的耐药性。结果表明,183株金黄色葡萄球菌对24种抗菌药物均呈现不同程度的耐药性,对常用的青霉素类、磺胺类药物及甲氧苄啶严重耐药,耐药率高达81%以上;其中检测出MRSA42株,检出率为22.95%。94.00%的金黄色葡萄球菌菌株呈现多重耐药,耐药谱型主要分布在3～8耐之间,以7耐菌株所占比例最大(15.30%),共有119种耐药谱型。     

鸡舍环境耐药细菌气溶胶及其向环境传播的研究

通过对舍内粪便、空气和舍外环境(10、50、100m)中的空气分离到的细菌鉴定，包括细菌DNA分子生物学检测及药物敏感实验以及细菌含量统计学分析，证明三者之间存在着内在联系；舍内环境微生物气溶胶包括耐药细菌通过舍内外气体交换排往鸡舍周围环境，舍周边环境在一定范围内受到生物污染。研究显示，在粪便、舍内空气、舍外环境10、50、100m空气中分离的金黄色葡萄球菌和链球菌80％以上对氯霉素(CMP)和对苯唑青霉素(OXA)耐药；粪便和舍内外环境空气中的80％以上的大肠杆菌和沙门氏菌对氨苄青霉素(AMP）、氯霉素(CMP)、头孢唑啉(CFZ)均为抑制。证明该鸡舍曾经使用过这些药物治疗或作为添加剂在饲料中添加，鸡群产生了耐药细菌菌株，并向环境传播。测量数据统计结果指出，十里河鸡场鸡舍内环境空气中的大肠杆菌、沙门氏菌、金黄色葡萄球菌和链球菌与舍外环境中相应菌群含量比较差异显著或极显著(P＜0．05或P＜0．01)，但舍外10、50、100m之间的相应菌群比较差异不显著(P＞0．05)，这说明鸡舍内环境细菌气溶胶含量产生多，含量高，菌源强度大；另一方面，反应了鸡舍内环境微生物气溶胶包括耐药细菌能向舍外环境扩散，位舍外环境在一定的范围内受到动物源性生物污染，影响范围大于100m。通过对鸡舍环境微生物气溶胶来源的探讨发现，粪便中的大肠杆菌、沙门氏菌、金黄色葡萄球菌和链球菌与舍内空气相应菌群数量关系比较，呈现正相关(r=0.99)。这从一个侧面揭示了粪便可能是微生物气溶胶的主要来源。     

Incidence of quinolone resistance in strains of Salmonella isolated from poultry, cattle and pigs in Germany between 1998 and 2001

This paper reports the susceptibility to the quinolone nalidixic acid and the fluoroquinolone ciprofloxacin of 14,514 strains of Salmonella isolated in Germany from poultry, cattle and pigs between 1998 and 2001. Quinolone-resistant salmonellae were most frequently isolated from poultry, with a prevalence of 10.2 to 16.8 per cent. Poultry-associated serotypes, such as Salmonella Paratyphi B (d-tartrate positive), Salmonella Hadar and Salmonella Virchow, had the highest prevalence of quinolone resistance, ranging between 35 and 74 per cent. All the nalidixic acid-resistant strains also had a reduced susceptibility to ciprofloxacin, with minimum inhibitory concentrations (MICS) of 0.125 to 2 microg/ml. A comparison of the MICS for ciprofloxacin of the strains of these poultry-associated serotypes and Salmonella Enteritidis phage type 4 isolated in 1998/99 and 2000/01 indicated that there had been a shift towards higher MIC values of up to 2 microg/ml. The quinolone resistance-determining region (QRDR) of the gyrA gene and the homologue region of the parC gene of 31 selected strains were sequenced. Several different amino acid changes were observed in gyrA of the quinolone-resistant isolates at positions 83 and 87, but no substitutions were observed in parC.     

The prevalence and PCR detection of Salmonella contamination in raw poultry

Contaminated poultry meat has been identified as one of the principal foodborne sources of Salmonella. The development of rapid detection assays for Salmonella would enable official agencies and food industries to identify contaminated foodstuffs in a more timely manner. In addition, these diagnostic tools could allow more 'real time' decisions to be made regarding end product acceptability. In this study, a survey was carried out to determine the prevalence of Salmonella in raw broiler carcasses. A total of 198 neck skin samples were obtained from within 40 flocks at a commercial broiler slaughtering facility. The presence of Salmonella was assessed by traditional culture methods and by a Salmonella-specific polymerase chain reaction (PCR) test. Salmonella was recovered from 32 (16%) of all samples using traditional culture methods. In contrast, the PCR assay proved to be more sensitive and detected Salmonella DNA in 38 (19%) of the samples tested. The pathogen was detected in 45 (23%) of the 198 samples when culture and PCR results were combined. The sensitivity of the PCR test was also greater than culture when detecting Salmonella from within flocks (53% of flocks by PCR, 30% of flocks by culture). The combination of both tests revealed that 55% of the flocks were contaminated with Salmonella. The PCR assay proved to be a highly specific and sensitive method for detecting Salmonella and the incorporation of a routine PCR test in conjunction with standard culture could be effective in providing a more accurate profile of the prevalence of this pathogen in broiler carcasses.     

Resistance to various antibiotics of Salmonella and Escherichia coli isolated from registrable farm feeds

Resistance to 20 antibiotics of 128 Salmonella and 97 Escherichia coli isolates from various registrable farm feeds was determined. A high frequency of comparatively low levels of resistance was found in both the Salmonella and E. coli isolates. This, together with an elevated frequency of multiple resistance, indicates that problems related to an effective transfer in bacterial populations of resistance to certain antibiotics are a distinct possibility. The addition of antibiotics, such as penicillin and tetracyclines, to animal feeds can create conditions for rapid selection amongst bacteria resistant to antibiotics. The numbers of resistant bacteria in the animal environment may be increased and may lead to the development of veterinary and human health problems from the possible transfer of antimicrobial resistance from animal pathogens to human pathogens or spreading in the human population of animal pathogens resistant to antibiotics. There is a need for caution in the use of antibiotics, particularly in animal feeds. Extended survey of, and epidemiological research on, farm feeds, manufacturing mills and animal production units are emphasized.