Phenotypes and genotypes of campylobacter strains isolated after cleaning and disinfection in poultry slaughterhouses

Campylobacter is responsible for human bacterial enteritis and poultry meat is recognised as a primary source of infection. In slaughterhouses, cleaning and disinfection procedures are performed daily, and it has been suggested that disinfectant molecules might select for antibiotic resistant strains if shared targets or combined resistance mechanisms were involved. The aim of the study was to investigate if cleaning and disinfection procedures in poultry slaughterhouses select for antibiotic resistance in Campylobacter jejuni and C. coli and to determine the genotypes of isolates collected after cleaning and disinfection. Nine sampling visits were made to four French slaughterhouses. Samples were collected from transport crates and equipment surfaces, before and after cleaning and disinfection. Minimal inhibitory concentrations of the recovered C. jejuni and C. coli isolates to six antibiotics and two disinfectants were measured. The C. jejuni isolates collected from equipment surfaces after cleaning and disinfection were subjected to PCR-RFLP typing. Twenty-five C. jejuni isolates and 1 C. coli were recovered from equipment surfaces after cleaning and disinfection during five visits to three different slaughterhouses. Those isolates did not show an increased resistance to the tested antibiotics compared to isolates collected before cleaning and disinfection. Only one or two genotypes were recovered after cleaning and disinfection during single visits to each slaughterhouse. This observation suggests that such genotypes may be particularly adapted to survive cleaning and disinfection stress. Understanding the survival mechanisms of Campylobacter should facilitate the implementation of better-targeted strategies and reduce the public health burden associated with Campylobacter infection.     

Resistance of Salmonella isolates in Germany

During 2000-2002 the National Veterinary Reference Laboratory for Salmonella (NRL-Salm) in Germany typed 11,911 isolates from animals, food, feed and the environment. All of them were tested for their susceptibility to 17 anti-microbial agents. Sixty-three per cent of all isolates were resistant and 40% were multiresistant (resistant against more than one anti-microbial). This general resistance level was strongly influenced by those specific serotypes which dominate the Salmonella epidemiology in Germany. Salmonella Typhimurium DT104 isolates from pig and cattle, and their resulting food products, were multiresistant in 98 and 94% of the cases respectively. During the period 2000-2003 an increasing quinolone resistance especially in Salmonella isolates from poultry and poultry meat (to 26%) and in S. Paratyphi B D-tartrate positive isolates (to 64%) could be observed. This increase was accompanied by a shift towards higher minimal inhibitory concentrations for ciprofloxacin.     

Possible associations between Salmonella persistence in poultry houses and resistance to commonly used disinfectants and a putative role of mar

A putative link between Salmonella persistence in the agricultural sector and resistance to disinfectants has been sparsely investigated. Therefore, minimum inhibitory concentration (MIC) tests against five disinfectants commonly used in poultry premises (formaldehyde, glutaraldehyde/benzalkonium chloride compound, oxidising compound, tar oil phenol, iodophor) were performed on 286 Salmonella isolates, including 256 from Danish broiler houses, altogether representing nine serotypes. Six of these isolates were used for adaptation and de-adaptation studies involving the five disinfectants. Amongst 60 of these isolates selected for growth studies in cyclohexane (possibly associated with up-regulated efflux), only one isolate grew. From this isolate and the six isolates used in the adaptation and de-adaptation studies, mutants highly resistant to triclosan (a disinfectant linked with mar-type resistance) were selected. In addition, adaptation and de-adaptation studies with triclosan were performed. For the 286 isolates, the small variations in MICs could not be associated with Salmonella persistence in Danish broiler houses or previous use of relevant disinfectants. Adaptation and de-adaptation did not alter MICs to the five farm disinfectants. Compared to the parent isolates, MICs for the triclosan adapted and de-adapted isolates and the triclosan mutants were significantly increased to triclosan, but not to the five disinfectants. Moreover, most of the triclosan adapted and de-adapted isolates grew in cyclohexane. Thus, there was no correlation between triclosan and cyclohexane resistance on one hand and resistance to the five disinfectants on the other, suggesting that triclosan resistance is not linked with resistance to these disinfectants.     

Antimicrobial susceptibility patterns of Salmonella isolates from cattle in feedlots

OBJECTIVE: To evaluate the antimicrobial susceptibility patterns of Salmonella isolates from feedlot cattle. DESIGN: Cross-sectional study. SAMPLE POPULATION: 263 Salmonella isolates. PROCEDURES: Fecal samples were collected from the floor of 2 pens in each of 100 feedlots. Two hundred eighty Salmonella isolates were recovered after bacteriologic culture from 38 pens. Of these, 263 isolates were available for antimicrobial susceptibility testing to 16 antimicrobials, using microbroth dilution breakpoint plates. RESULTS: Less than 5% of isolates were resistant to any of the antimicrobials tested, with the exception of sulfamethoxazole (15; 5.7%) and tetracycline (61; 23.2%). Most isolates (197; 74.9%) were susceptible to all antimicrobials tested, whereas 18 (6.8%) were resistant to 2 or more antimicrobials. The percentage of isolates with resistance to any antimicrobial varied by serotype. The percentage of isolates resistant to various antimicrobials was not related to concurrent use of antimicrobials in the feed. CONCLUSIONS AND CLINICAL RELEVANCE: With the exception of tetracycline and sulfamethoxazole, resistance of Salmonella isolates to any of the antimicrobials was uncommon. Most isolates were susceptible to all antimicrobials tested. Antimicrobial resistance was not related to the presence of antimicrobials in the ration being fed at the time of sample collection.     

Investigation of resistance of bacteria from commercial poultry sources to commercial disinfectants

Concern by consumers about food safety has resulted in increased pressure on poultry companies to develop effective sanitation programs. Salmonella isolates in hatcheries are often the same species isolated from processing plants. Resistance develops in bacteria after prolonged exposure to disinfectants. The methods available in published literature to detect the efficacy of disinfectants are labor intensive and do not consider how bacteria behave when adhered to a solid surface. We used a recently developed technique, which utilizes the actual surfaces on which the disinfectant is to be applied, to evaluate the degree of resistance to four commercially available disinfectants of 17 bacterial isolates from poultry hatcheries. We found that bacterial isolates within the same genus and species have different sensitivities to the same disinfectant. In addition, disinfectants with similar but not identical chemical formulations have different efficacies against the same bacteria.     

Inactivation of canine parvovirus by disinfectants and heat

The resistance of canine parvovirus (CPV) to inactivation by chemical and thermal means was investigated. CPV with and without extraneous protein was tested against 15 commonly used disinfectants at room temperature. The titres of CPV remaining after 5 minutes disinfection time were greater than or equal to those remaining after 30 minutes regardless of the presence or absence of protein. Disinfection of CPV was accomplished more quickly and at lower concentrations of disinfectants when extraneous protein was absent. Halogen compounds, aldehydes and sodium hydroxide were the most acceptable CPV disinfectants. The presence of protein interfered with the two most frequently recommended CPV disinfectants—sodium hypo-chlorite and formaldehyde. The resistance of CPV to inactivation by heat depended on the temperature and the duration of heating. Boiling (100°C) rapidly inactivated CPV. The virus was more resistant to lower temperatures. Detectable infectivity remained after 7 hours at 80°C and 72 hours at 56°C. At both 37°C and room temperature there was no significant change in infectivity titres over 72 hours.     

鱼塘生态体系中大肠埃希菌和沙门菌的耐药性研究

为了解鱼塘生态体系大肠埃希菌和沙门菌的耐药情况,从广东省佛山某鱼塘随机采集草鱼及生活圈的水土样品中(包括鱼肠内容物、鱼塘底泥、鱼塘水)分离出47株大肠埃希菌和23株沙门菌.采用纸片扩散法(Kirby-bauer,KB)对分离菌株进行了15种抗生素的敏感性试验,结果显示100%的菌株均耐1种及1种以上的抗生素多表现出对氨...     

食品动物源沙门氏菌质粒介导喹诺酮类耐药基因的检测与分析

为了解食品动物源沙门氏菌质粒介导喹诺酮类耐药性(Plasmid-mediated quinolone resistance,PMQR),采用微量肉汤稀释法和PCR方法,检测了316株食品动物源沙门氏菌对20种抗菌药物的敏感性,以及菌株中PMQR基因的携带率.结果显示:316株沙门氏菌对20种抗菌药物呈不同程度的耐药性,95.57％菌株为多重耐药菌;316株菌中未检出qnrA、qnrC、qnrD、qnrS和qepA基因,7.91％菌株检出qnrB基因,15.19％菌株检出aac(6 ′ )-Ib-cr基因,7.91％菌株检出oqxA基因,8.86％菌株检出oqxB基因,这是首次在沙门氏菌中发现oqxAB基因;98.11％PMQR阳性菌同时携带2种及以上的耐药基因,呈8～17耐的多重耐药性,其中以qnrB和aac(6′)-Ibcr基因型为主;53株PMQR阳性菌分属于5种不同的基因型,耐药表型或耐药基因型不同的菌株却有相同的PFGE谱型.本次检测的316株食品动物源沙门氏菌耐药较为严重;菌株主要携带qnrB、aac(6 ′ )-Ib-cr及oqxAB基因;不同来源菌株存在同一耐药克隆株的流行.     

Susceptibilities to antimicrobials and disinfectants in Salmonella isolates obtained from poultry and swine in Thailand

Salmonella enterica isolates from poultry (n=125) and swine (n=132) in Thailand were investigated for antibiotic resistance, susceptibility to disinfectants (benzalkonium chloride (BKC), chlorhexidine digluconate (CHX), zinc chloride and copper sulfate) and cyclohexane tolerance. Forty-two percent were of multiple resistance to antibiotics. The minimum inhibitory concentrations (MICs) of all antibiotics against isolates from swine were higher than that against the isolates from poultry. There were generally few variations in MICs to all disinfectants, indicating that the isolates had either no or only a limited degree of developed resistance to the disinfectants tested. Only 5 isolates (1.9%) were tolerant to cyclohexane. The proton-dependent efflux systems did not play a major role in the reduced susceptibility to BKC and CHX, since susceptibility was not restored when an efflux inhibitor, carbonyl cyanide m-chlorophenylhydrazone (CCCP) was present. Successive exposure to subinhibitory concentrations of BKC and CHX generated mutants resistant to BKC and CHX. A spontaneous BKC-resistant derivative expressed cross-resistance to antibiotics, chloramphenicol and erythromycin. The mechanism responsible for cross-resistance between BKC and antibiotics was not driven by the proton motif force (PMF).     

Characterization of antibiotic-resistant bacteria in rendered animal products.

Antibiotics are used in food animal production to treat diseases and also to improve performance. Antibiotics are not used on all farms, and antibiotic resistance is occasionally found on farms that do not use antibiotics. Rendered animal protein products are often included in poultry feeds and could potentially serve as a source of antibiotic-resistant bacteria. One hundred sixty-five rendered animal protein products from cattle, poultry, and fish were aseptically collected from poultry feed mills. Fifty-five percent of the poultry meal samples had detectable levels of gram-negative bacteria ranging from 40 to 10,440 colony-forming units/g of sample. Poultry meal and meat and bone meal had the greatest number of samples with bacteria resistant to five or more antibiotics. A high percentage of feed samples (85%) contained bacteria resistant to amoxicillin, ampicillin, clavulanic acid, or cephalothin, whereas few samples contained bacteria resistant to ciprofloxacin, kanamycin, or trimethoprim/sulfamethoxazole. Acinetobacter calcoaceticus, Citrobacter freundii, and Enterobacter cloacae were the most commonly isolated antibiotic-resistant bacteria. Isolation for Salmonella was also performed, with 14% of the meat and bone meal samples containing Salmonella sp. Only one of the meat and bone meal isolates, Salmonella livingstone, was resistant to five or more antibiotics. Many of the antibiotic-resistant bacteria contained integrons, genetic elements that mediate multiple drug resistance.     

禽源致病性沙门氏菌耐药表型与耐药基因的分析

为了解江苏苏中地区禽源沙门氏菌耐药现象及耐药基因的存在情况,采用K-B法对22株沙门氏菌分离株进行12种抗菌药物敏感性试验,并通过PCR方法检测了12种耐药基因的存在情况。结果显示,所有菌株对甲氧胺嘧啶、阿米卡星、庆大霉素、萘啶酮酸、诺氟沙星的耐药率均在72.73%～100%之间;存在着5种耐药表型,每个耐药表型均在4重或4重以上,耐药最多菌株可耐受10种药物;扩增出9种耐药基因,与GenBank中的参考序列同源性均在98%以上;耐药表型与耐药基因的存在基本一致,但无绝对相关性。本研究结果表明苏中地区沙门氏菌耐药现象非常严重,且耐药基因广泛存在。     

Detection of Salmonella organisms and assessment of a protocol for removal of contamination in horse stalls at a veterinary teaching hospital

OBJECTIVES: To assess methods of detecting environmental contamination with Salmonella organisms and evaluate a cleaning and disinfection protocol for horse stalls in a veterinary teaching hospital. DESIGN: Original study. SAMPLE POPULATION: 37 horses with diarrhea likely to be caused by Salmonella infection and their stall environments. PROCEDURES: Fecal samples were collected from horses daily during hospitalization; samples were obtained from stall sites after cleaning and application of disinfectants. Fecal and environmental samples were cultured for Salmonella spp and tested via polymerase chain reaction (PCR) assay to detect Salmonella DNA. RESULTS: 1 horse died and 2 were discharged prior to sample collection. Fecal samples from 9 of 34 horses yielded growth of Salmonella organisms on bacteriologic culture, and 23 yielded positive results via PCR assay on > or = 1 occasion. Among environmental samples from 21 stalls, salmonellae were detected at > or = 1 stall site on 6 of 78 occasions, and > or = 1 stall site yielded positive results via PCR assay on 69 of 77 occasions. Salmonella DNA was detected more frequently in samples of stall drains, cracks, and corners. Salmonella spp were cultured from samples of 3 stalls after both initial and second cleaning and disinfection cycles, but no organisms were detected in samples obtained after use of a peroxygen disinfectant. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that stalls in which horses with salmonellosis were housed should only be used to accommodate newly hospitalized horses after samples (collected after 2 cycles of cleaning and disinfection) from drains, cracks, and corners yield negative results on bacteriologic culture.     

Characterization of Salmonella Dublin and Salmonella Typhimurium (Copenhagen) Isolates from Cattle

Brackelsberg, C.A., Nolan, L.K. and Brown, J., 1997. Characterization of Salmonella dublin and Salmonella typhimurium (Copenhagen) isolates from cattle. Veterinary Research Communications, 21 (6), 409-420Eight Salmonella typhimurium (Copenhagen) and eight Salmonella dublin isolates from cattle were compared by their antibiotic resistance patterns, by their production of colicin, aerobactin, haemolysin and capsule, by their possession of transmissible R plasmids and the spvC gene, and by their ability to invade and replicate within cultured epithelial cells. The two groups differed in their antibiotic resistance profiles, with more of the host-adapted S. dublin isolates resistant to tetracycline than were the non-host-adapted S. typhimurium (Copenhagen) group, but more of the S. typhimurium (Copenhagen) isolates resistant to the other antibiotics tested. None of the isolates produced colicin, but all produced aerobactin. One isolate in each group was encapsulated. All of the S. typhimurium (Copenhagen) and S. dublin isolates contained plasmids, and all of them contained the spvC-homologous sequences. Four of the S. typhimurium (Copenhagen) isolates were able to transfer an R plasmid to a recipient organism by conjugation. One of the five S. dublin isolates, which showed resistance to some of the antibiotics tested, was able to transfer an R plasmid by conjugation. Both groups of isolates invaded cultured epithelial cells to a similar degree after 1 h, but the S. dublin isolates reached significantly higher levels within the cells than did S. typhimurium (Copenhagen) after 9 h. This ability may, in part, explain the association of S. dublin with more severe forms of salmonellosis and prolonged carrier states. Further study of the intracellular growth of these isolates seems warranted.     

鸡白痢沙门菌的分离鉴定与耐药性分析

本研究采集某鸡场疑似感染鸡白痢的病死鸡组织,进行了沙门菌的分离、鉴定,以及对10种常见抗生素的药物敏感性试验.结果显示:分离株均为鸡白痢沙门菌,对阿米卡星(0％)、庆大霉素(0％)、头孢噻肟(3.70％)的耐药率较低,对链霉素(88.89％)、四环素(62.96％)、阿莫西林(51.85％)的耐药率较高;分离株至少可对...     

The role of disinfectant resistance of Salmonella enterica serotype enteritidis in recurring infections in Pennsylvania egg quality assurance program monitored flocks

The Pennsylvania egg quality assurance program (PEQAP) has made major gains in the reduction of Salmonella enterica serotype enteritidis (S. enteritidis). However, S. enteritidis continues to be a major food safety concern for the commercial egg laying industry. Despite intensive control efforts through PEQAP, some commercial egg layer houses still remain positive for S. enteritidis. The primary objective of this study was to determine whether S. enteritidis isolates obtained from historically environmentally S. enteritidis-positive houses were resistant to commonly used disinfectants. Archived S. enteritidis isolates (environmental, rodent, or egg) were compared with recently obtained isolates from the environment, rodents, or eggs from the same S. enteritidis-positive house. In addition, the isolates were compared with archived isolates from those premises that appeared to have eliminated S. enteritidis from their layer facilities. The official methods of the use-dilution analysis of the Association of Official Analytical Chemists were used to evaluate each disinfectant product. Two phenolic, one quaternary ammonium, and one combination product containing quaternary ammonium and formaldehyde were evaluated, in addition to one sodium hypochlorite detergent. All products diluted according to the manufacturers' recommendations killed the S. enteritidis isolates in this test system. There was no difference in susceptibility or resistance to the disinfectants used between the isolates from those facilities that remained S. enteritidis-positive and those that appeared to have eliminated S. enteritidis from their facility.     

Identification of sources of Salmonella organisms in a veterinary teaching hospital and evaluation of the effects of disinfectants on detection of Salmonella organisms on surface materials

OBJECTIVE: To determine sources of Salmonella organisms in a veterinary teaching hospital, compare bacterial culture with polymerase chain reaction (PCR) testing for detection of Salmonella organisms in environmental samples, and evaluate the effects of various disinfectants on detection of Salmonella organisms on surface materials. DESIGN: Prospective study. SAMPLE POPULATION: Fecal samples from 638 hospitalized horses and 783 environmental samples. PROCEDURE: Standard bacterial culture techniques were used; the PCR test amplified a segment of the Salmonella DNA. Five disinfectants were mixed with Salmonella suspensions, and bacterial culture was performed. Swab samples were collected from 7 surface materials after inoculation of the surfaces with Salmonella Typhimurium, with or without addition of a disinfectant, and submitted for bacterial culture and PCR testing. RESULTS: Salmonella organisms were detected in fecal samples from 35 (5.5%) horses. For environmental samples, the proportion of positive bacterial culture results (1/783) was significantly less than the proportion of positive PCR test results (110/783), probably because of detection of nonviable DNA by the PCR test. Detection of Salmonella organisms varied with the surface material tested, the method of detection (bacterial culture vs PCR testing), and the presence and type of disinfectant. CONCLUSIONS AND CLINICAL RELEVANCE: Results of the present study suggested that Salmonella organisms can be isolated from feces of hospitalized horses and a variety of environmental surfaces in a large animal hospital. Although recovery of Salmonella organisms was affected by surface material and disinfectant, bleach was the most effective disinfectant on the largest number of surfaces tested.     

Sources of Salmonella contamination during broiler production in Eastern Spain

Prevention of Salmonella contamination of poultry products requires detailed knowledge of the main sources associated with its presence in the production system. The aims of this study were to determine the main sources of Salmonella contamination in broiler production during growing, to assess the risk factors for Salmonella contamination at the end of the rearing period and to determine the main serovars involved in broiler production systems in Eastern Spain. A total of 65 different broiler houses from different farms were sampled. Each house was sampled at different times during the rearing period. First, when the previous flock was taken to the slaughterhouse, samples of dust, surfaces and previous flock faeces were collected. After cleaning and disinfection (C&D), samples of dust and surfaces were also taken. On the first day of rearing, samples of water, bedding, farming boots, meconiums, delivery-box liners and feed were collected. During rearing, feed samples were taken directly from the truck and from feeders. On slaughter day, samples of dust, surfaces, water, feed and faeces were also collected. Finally, two days after slaughter, carriers (rodents, flies and beetles) were trapped. All samples collected were analysed according to ISO 6579:2002 (Annex D) and positive samples were serotyped in accordance with Kauffman-White-Le-Minor technique. Our results showed that all different types of samples collected were contaminated with Salmonella (prevalence ranged between 1.5% and 38.6%). The most contaminated samples related with poultry production were: delivery-box liners (32.0%), faeces samples (31.2%), dust samples (25.0%), farming boots (19.7%) and feed from feeders (16.0%). However, the most important risk factors for Salmonella contamination of the flocks at the end of the rearing period were Salmonella status of the house after cleaning and disinfection, Salmonella status of day-old chick flocks and feed from feeders. Twenty-one different serovars were isolated from the samples analysed. The most prevalent were in decreasing order: Salmonella Enteritidis (52.9%), S. Hadar (17.8%), S. Virchow (8.9%) and S. Ohio (5.4%). The study suggested that there are many sources for Salmonella contamination and persistence in broiler production. Hence, the whole production chain needs to be controlled to eradicate the bacteria from primary production.     

Outbreak of Salmonella Dublin-associated abortion in Danish fur farms

Outbreaks of Salmonella Dublin infections were recorded in 25 Danish mink and fox farms. All farms suffered extensive disease problems; clinical and pathological observations included abortion, stillbirths, necrotizing endometritis, and increased mortality. By genotyping with pulsed-field gel electrophoresis and amplified fragment length polymorphism, all isolates of S. Dublin had indistinguishable patterns. The outbreaks took place during April and May, around the time of whelping. During this period, mink are particularly susceptible to Salmonella infections. All affected farms were served by the same feed factory and it was concluded that a batch of contaminated feed was responsible for the outbreaks, although repeated culture of feed samples collected during the same period were negative. No other likely source could be identified. The results emphasize the importance of strict hygiene measures at feed factories and the proper use of ingredients of known Salmonella status, in particular during the whelping season. Infected mink farms did not have a higher risk of outbreak of salmonellosis in the year following the outbreak.     

Antimicrobial resistance in Salmonella enterica serovar Dublin isolates from beef and dairy sources

Salmonella enterica serovar Dublin (S. Dublin) is a cattle-adapted Salmonella serovar, so if antimicrobial resistance in S. Dublin arises as a result of antimicrobial use this most likely occurs within the cattle reservoir without impact from antimicrobial use in humans. We tested the antimicrobial resistance of bovine-origin S. Dublin isolates from 1986 through 2004 using a standard disk diffusion method. High proportions of isolates throughout the time period were resistant to one or more antimicrobials, and a marked increase in resistance to ceftazidime occurred between 2000 and 2004. Dairy-origin isolates were more likely to be resistant to several antibiotics than were isolates from beef operations where exposure to antimicrobials is likely to be less frequent. Plasmid analysis of a subset of isolates also supported the hypothesis that antimicrobial resistance traits in the cattle-adapted serovar Dublin were acquired within the bovine host environment.     

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.