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1.
This study aims to investigate the genetic diversity of thermotolerant Campylobacter in commercial broiler flocks and in the environment of broiler farms in Belgium. Seven out of 18 investigated flocks became colonized during rearing. Fluorescent amplified fragment length polymorphism (FAFLP), pulsed field gel electrophoresis (PFGE), restriction fragment length polymorphism of the flagellin A gene (flaA-RFLP) and antimicrobial resistance profile (ARP) were used for typing of the isolates. By the combination of FAFLP and PFGE, 22 Campylobacter genotypes could be distinguished. Colonization was almost exclusively with Campylobacter jejuni and unique genotypes were found in each flock. Multiple genotypes were detected in the broilers of 3 flocks, either simultaneously or successively. In 5 flocks, strains that were resistant to at least one antibiotic (mostly tetracycline) were found. The presence of other broiler houses on the farm did not result in a higher probability of colonization. The nipple water was contaminated with the same genotype as the broilers, illustrating its importance for transmission of Campylobacter. The same genotype was detected in a water puddle and in the broiler flock during rearing in 3 flocks. Once, the same genotype was isolated from the ditch water shortly before it was detected in the broilers.  相似文献   

2.
We investigated the genotype diversity and dynamics of Campylobacter in a commercial broiler flock during rearing and slaughter. In total, 220 Campylobacter jejuni isolates collected on four sampling occasions during rearing and from routine sampling during slaughter were subtyped by SmaI macrorestriction and pulsed-field gel electrophoresis, PFGE. Eight different SmaI types were found. During rearing, a subsequent addition of genotypes occurred, with two SmaI types found at 2 weeks of age and six types on the day before slaughter. All types that were detected in more than one isolate were also found on all succeeding sampling occasions, including the slaughter sampling. Two new types were found in the slaughter samples. In two-thirds of the individual birds sampled the day before slaughter, more than one SmaI type were found, although there was a clear tendency for dominance of one type in individual birds. Our results show that multiple genotypes of C. jejuni may be present in a commercial broiler flock during rearing and even in gastrointestinal tracts of individual birds. Both recurring environmental exposure and genetic changes within the population may explain the genotype diversity. Although the distribution of genotypes varied between different sampling occasions, we found no indication that any subtype excluded another during the rearing of the broiler flock.  相似文献   

3.
Screening of broiler flocks for their Campylobacter carriage on farm level and consequently the spread of Campylobacter spp. during slaughtering can help to identify hygiene control points. Therefore, between December 2001 and August 2002 in total 51 broiler flocks from three farms of different geographical regions in Germany were analysed for thermophilic Campylobacter. Campylobacter spp. were isolated from 45% of the broiler flocks examined. Subsequently, 1101 samples were taken from 22 flocks during different stages of processing. Samples were collected from: transport crates before and after cleaning/disinfection, evisceration, post-scalded and post-chilled carcasses and endproducts. Additionally, 45 selected Campylobacter isolates of droppings were genotyped by pulsed-field gel electrophoresis (PFGE). Campylobacter carriage of flocks showed seasonal variation, with the highest contamination rate during the period of June to August. No evidence was found for a horizontal transmission from one broiler flock to the next via a persistent house-contamination. In each positive flock, one to three different genotypes were found. One or two clones dominated isolations obtained from the farm level. The fact that in different flocks indistinguishable isolates of clonal origin were detected during the same rearing period suggested a transmission between the broiler flocks or an intermittent common external source. In one case, isolates of clonal origin were detected in various farms during different rearing periods. Sampling during processing confirmed that the entrance of a positive flock resulted in contamination of the abattoir environment. Campylobacter spp. were isolated from all sampling stages along the processing line, with a percentage of 91.1-100 of isolates at different stages of slaughtering.  相似文献   

4.
The carry-over of Campylobacter strains from one flock to a subsequent flock in the same broiler house has been studied using molecular epidemiological techniques. In all, 524 Campylobacter strains, isolated from two sequential broiler flocks from 60 broiler houses, were typed by restriction fragment polymorphism of the polymerase chain reaction (PCR) product of the flaA and flaB genes (fla typing). Selected strains were also typed using pulsed field gel electrophoresis (PFGE). By fla typing, 15 (21%) of the 60 houses with Campylobacter-positive sequential flocks had identical genotypes. In 10 (16% overall) of these houses the strains were also identical by PFGE. The difference in PFGE patterns in the strains from the three remaining houses may be indicative of genetic instability. Overall, these results suggest that carry-over from one flock to a subsequent flock in the same house is a relatively infrequent event and, therefore, that routine broiler house cleansing and/or disinfection is largely adequate to eliminate Campylobacter contamination. An alternative explanation of the low level carry-over is a persistent source or reservoir, external to the environment of the broiler houses.  相似文献   

5.
Campylobacter is recognized as one of the leading cause of gastroenteritis worldwide, and is frequently isolated from the small intestines and ceca microflora of chickens. Twenty-one out of 81 Campylobacter-positive poultry flocks were selected to evaluate the genetic diversity of Campylobacter isolates and to study the distribution of genotypes among flocks. Campylobacter isolates recovered from chicken carcasses and ceca were analyzed by pulsed-field gel electrophoresis (PFGE). Little diversity was found among Campylobacter strains isolated from a given carcass, with a maximum of 2 different genotypes being present. However, at flock level, as many as 4 different profiles were observed. Typing of strains showed that most strains isolated from ceca were similar to those isolated from corresponding broiler carcasses. A total of 39 different macrorestriction profiles were observed, with evidence of Campylobacter cross-contamination among broiler flocks in Quebec slaughterhouses. Surprisingly, some flocks shared related genotypes both with and without sharing similar rearing practices. Existence of such cross-contamination must be considered to in developing strategies to control Campylobacter in chickens, and to avoid bacteria contamination of noncolonized flocks. Further typing studies of Campylobacter found in hatcheries, farm environment, and crates or trucks in Quebec might be helpful in elucidating the kinetics of broiler chicken Campylobacter contamination.  相似文献   

6.
The aim of this study was to analyse the genetic diversity among Clostridium perfringens isolates from Danish broiler chickens since both sick and presumably healthy animals were investigated. Isolates (n=279) collected from chickens from 25 farms were analysed by pulsed-field gel electrophoresis (PFGE) with the restriction enzyme SmaI. A high genetic diversity was found. Isolates with different PFGE types were toxin typed by PCR and all were found to be of type A. The results showed that healthy broiler chickens carried several different C. perfringens clones both within a flock and even within individual birds, whereas flocks suffering from necrotic enteritis (NE) or cholangio-hepatitis carried only one or two clones.  相似文献   

7.
To investigate the effects of rearing practices of commercial broiler chickens on the incidence of antimicrobial resistance in commensal Escherichia coli isolates, fecal E. coli isolates obtained in 4 farms were screened for anitimicrobial resistance. Ten E. coli isolates were recovered from each of the fecal samples collected from 10 birds in the farms at the ages of 2 days, 14-17 days, and 47-50 days. In 2 out of the 4 farms, no antimicrobials were used during the rearing period. In the other two farms, following collection of the fecal samples at 14 and 15 days of age, oxytetracycline (OTC), sulfadimethoxine (SDMX), and tylosin were given to birds on one farm and SDMX was used in the other. Isolates resistant to ampicillin and OTC that were obtained from an untreated flock at different sampling times were closely related to each other by pulsed-field gel electrophoresis patterns (PFGE) of XbaI-digested chromosomal DNA. PFGE analysis together with in vitro conjugation experiments suggested that diversity of resistance phenotypes within a clone may be resulted from the acquisition and loss of R-plasmids in an untreated and a treated flock. The numbers of resistance phenotypes observed among fecal isolates increased during the growth of the chickens in all the farms. The results in the present study suggest that persistence of commensal E. coli strains resistant to antimicrobials even in the absence of antimicrobial administration. It is also hypothesized that horizontal transmission of resistance determinants resulted in the emergence of different resistance phenotypes in those farms.  相似文献   

8.
A nation-wide survey was carried out in 370 randomly chosen French commercial broiler chicken flocks from October 2005 to September 2006 to determine Salmonella spp. prevalence and to identify risk factors for contamination, at the end of the rearing period. The Salmonella status of the flocks was assessed from five faecal samples (litter swabs) analysed by classical bacteriological methods. A flock with at least one contaminated sample was classified as a Salmonella-positive flock. The apparent prevalence of Salmonella was 8.6% (95% CI: 5.7, 11.5%). The most prevalent serovar was S. hadar followed by S. anatum and S. mbandaka. Logistic regression methods were used to analyse the associations between husbandry practices, farm characteristics, general hygiene and the Salmonella status of the sample. The risk for Salmonella contamination of the flock at the end of the rearing period increased when neighbours helped in the placement of day-old chicks. On the contrary, the risk decreased when mobile equipment was dismantled before cleaning and disinfection, when the farm had a specific container for dead-bird disposal and when acetic acid was added to the drinking water.  相似文献   

9.
NetB toxin from Clostridium perfringens is a major virulence factor in necrotic enteritis in poultry. In this study the efficacy of NetB as a vaccine antigen to protect chickens from necrotic enteritis was examined. Broiler chickens were immunized subcutaneously with purified recombinant NetB (rNetB), formalin treated bacterin and cell free toxoid with or without rNetB supplementation. Intestinal lesion scores and NetB antibody levels were measured to determine protection after mild oral gavage, moderate in-feed and heavy in-feed challenges with virulent C. perfringens isolates. Birds immunized with rNetB were significantly protected against necrotic enteritis when challenged with a mild oral dose of virulent bacteria, but were not protected when a more robust challenge was used. Bacterin and cell free toxoid without rNetB supplementation did not protect birds from moderate and severe in-feed challenge. Only birds immunized with bacterin and cell free toxoid supplemented with rNetB showed significant protection against moderate and severe in-feed challenge, with the later giving the greatest protection. Higher NetB antibody titres were observed in birds immunized with rNetB compared to those vaccinated with bacterin or toxoid, suggesting that the in vitro levels of NetB produced by virulent C. perfringens isolates are too low to induce the development of a strong immune response. These results suggest that vaccination with NetB alone may not be sufficient to protect birds from necrotic enteritis in the field, but that in combination with other cellular or cell-free antigens it can significantly protect chickens from disease.  相似文献   

10.
The efficacy of narasin in the control of necrotic enteritis (NE) was investigated in a floor pen study of 2000 broiler chickens using a Clostridium perfringens feed inoculum challenge model. Treatments were 1) nonmedicated, nonchallenged; 2) nonmedicated, challenged; 3) narasin, nonchallenged; 4) narasin, challenged. Narasin was administered at 70 ppm in the feed from day 0 to trial termination on day 41. Challenge inoculum contained approximately 1 x 10(8) colony-forming units CP/ml and was administered from day 14 to day 16. In the unmedicated groups, challenged birds had significantly (P < 0.05) lower mean body weight and reduced feed efficiency at day 21 and significantly (P < 0.01) higher cumulative NE mortality at day 41 compared with unchallenged. Similarly, among unmedicated birds, those challenged had a significantly (P < 0.01) higher mean NE score on day 17 and significantly (P < 0.05) higher mean huddling scores on days 15-17 than unchallenged. Among challenged birds, those fed narasin had significantly (P < 0.05) higher mean body weight and improved feed efficiency at days 21 and 41 and significantly (P < 0.01) lower cumulative NE mortality at day 41 than unmedicated. Similarly, among challenged birds, those receiving narasin had a lower mean NE score on day 17 (P > 0.05) and significantly (P < 0.05) lower huddling scores on days 16 and 17 than unmedicated. Coccidiosis lesion scores were zero for birds euthanatized from all treatment groups on day 17, suggesting that the beneficial effects of narasin were not due to prevention of coccidiosis. This study thus provides evidence that narasin is effective in the prevention of necrotic enteritis in broiler chickens.  相似文献   

11.
In Denmark, increased problems associated with streptococci and enterococci have been observed in broilers and broiler parent flocks, resulting in increased mortalities, uneven flocks, and subsequent downgrading and increased condemnations. Postmortem lesions associated with recent outbreaks due to Enterococcus hirae have been accompanied or dominated by septicemia and endocarditis. As a result of infection at an early age and relatively low mortality rates, outbreaks are not always clearly defined and may go unnoticed or may be attributed to poor chick quality. For the same reasons, the pathogenesis and epidemiology of observed outbreaks has only remained speculative. Four separate outbreaks associated with E. hirae infections in broiler flocks occurring between 1998 and 2002 have been investigated. Two of the outbreaks indicated evidence of two separate clones, with 89% and 79% of isolates involved in the individual outbreaks belonging to a single pulsed-field gel electrophoresis (PFGE) profile, respectively. Another outbreak (outbreak 4) demonstrated clear clonality, with all isolates demonstrating affinity to one of two PFGE profiles that differed by only two bands. However, all three outbreaks demonstrated a different clone. The remaining outbreak was nondonal, with isolates distributed between six separate PFGE profiles. One of the outbreaks (outbreak 4) was descended from a parent broiler flock previously associated with an outbreak of Streptococcus gallinaceus; the flock also exhibited septicemia and endocarditis. Initial indications suggested the possibility of vertical transmission of S. gallinaceus to the current broiler flock, causing infection. By extended phenotypic characterization and subsequent genetic characterization, including 16S rRNA sequencing, all strains from the four outbreaks were confirmed as E. hirae. This investigation highlights the problems associated with characterizing enterococci infections in broiler flocks.  相似文献   

12.
The dosing of young chicks with cultures of normal gut flora has been termed "competitive exclusion" (CE). This study was undertaken to examine, under field conditions, the effect of CE treatment on counts of intestinal Clostridium perfringens (CP) and on the occurrence of CP-associated disease in broiler chickens. A farm having recurrent CP-associated health problems was selected as study site. The study comprised four broiler houses, with one treated and one untreated flock per house. Treated birds were sprayed with the CE product Broilact upon arrival at the farm. All flocks were offered feed containing the ionophorous anticoccidial agent narasin. The feed did not contain growth promoters. Treatment was associated with positive but statistically nonsignificant effects on gut health. Delayed intestinal proliferation of CP and delayed appearance of CP-associated gut lesions were found in CE-treated flocks. This delay was associated with improved production performance at slaughter.  相似文献   

13.
Two trials were conducted to study the effects of a competitive exclusion (CE) product BROILACT and the anticoccidial narasin on the incidence of necrotic enteritis (NE), the numbers of Clostridium perfringens (CP) in the caeca of broiler chicks and the performance of the birds. In trial 1 the effects of type of protein and partial replacement of a narasin containing diet with whole wheat were also studied. All groups of chicks were studied up to the point of slaughter at 43 days of age and after evisceration in a processing plant to determine slaughter yield. In trial 1, statistically significant results included the following: CE-treatment reduced total mortality, and incidence of NE, on diet containing animal but not vegetable protein. Caecal carriage of CP was also reduced, while slaughter yield increased. Narasin reduced caecal carriage of CP and increased both growth rate and slaughter yield in both trials. Whole wheat replacement improved feed conversion but reduced bird growth rate. In trial 2, both CE-treatment and narasin influenced feed intake, CE-treatment significantly only at days 22 and 44. Narasin improved feed conversion until 5 weeks of age and CE-treatment did so until 22 days of age. In both trials, there was also an interaction effect indicating that CE-treatment increased slaughter yield for birds that were not fed narasin.  相似文献   

14.
We inoculated broiler chicks with mixtures of Clostridium perfringens strains to investigate the single strain dominance observed in natural cases of necrotic enteritis (NE) [Nauerby, B., Pedersen, K., Madsen, M., 2003. Analysis by pulsed-field gel electrophoresis of the genetic diversity among Clostridium perfringens isolates from chickens. Vet. Microbiol. 94, 257-266]. Pre-inoculation bacteriologic culture of chick intestines yielded up to six pulsed-field gel electrophoresis (PFGE) types of C. perfringens. Birds developed typical NE lesions in response to administration (2x per day for 4 days) of a combined inoculum comprising one NE strain (JGS4143, PFGE pattern 8) and four non-NE strains (from piglet necrotizing enteritis, chicken normal flora, human gas gangrene, and bovine neonatal enteritis). After inoculation commenced, only the NE strain was recovered through the first post-inoculation day, in spite of intense efforts to recover pre-challenge flora strains and the other challenge strains. Thereafter, pre-inoculation and previously undetected PFGE types were found, and JGS4143 became undetectable. Birds inoculated simultaneously with five NE strains (from disease in chickens or turkeys, and including JGS4143) also developed lesions, but again only JGS4143 was recovered through the 1st day post-challenge. At that time, birds began to be repopulated with pre-challenge PFGE types. Two NE strains (JGS4143 and JGS4064) produced bacteriocins, which inhibited each other and normal flora strains (n=17), while normal flora strains inhibited neither NE strains nor each other. Thus, it appears that naturally occurring dominance of the gut by NE strains can be reproduced experimentally. Bacteriocins directed against normal flora could possibly provide the necessary advantage, although inhibition of one NE strain by another suggests that other factors may be partially or completely responsible for the dominance.  相似文献   

15.
A cross-sectional study was conducted on a commercial egg-producing farm with a history of wet litter. A total of 600 fresh caecal faecal samples were obtained from under cages of laying hens in three sheds each containing flocks of approximately 5400 hens. Samples were cultured for intestinal spirochaetes, and growth on the primary isolation plate was observed under a phase contrast microscope and subjected to PCRs specific for the intestinal spirochaetes Brachyspira intermedia and Brachyspira pilosicoli. Spirochaete isolates obtained in pure culture were assessed for their ability to cause haemolysis on blood agar and to produce indole, and were typed using pulsed field gel electrophoresis (PFGE). A 1250 base pair portion of the 16S rRNA gene of three B. intermedia and five unidentified isolates was sequenced, and the sequences compared with those of other Brachyspira species. Overall, 121 (20.2%) of the faecal samples contained spirochaetes as determined by growth on the plate and microscopy. Using PCR on the primary growth from these positive samples, 43 (7.2% overall) were shown to contain B. intermedia, 8 (1.3%) to contain B. pilosicoli, and 70 (11.7%) were PCR negative. Only 24 isolates of B. intermedia and five isolates of unknown species were obtained in pure culture. Comparative analysis of the 16S rRNA gene sequence identified the non-B. intermedia isolates as belonging to the proposed species "Brachyspira pulli". PFGE analysis of the B. intermedia strains identified them as having four major banding patterns. Individual patterns were found in hens from different flocks, suggesting cross-transmission of strains between flocks. No environmental sources of infection were identified. The youngest flock had a significantly lower level of colonisation with B. intermedia than the flock of intermediate age (P = 0.004), suggesting that following initial infection of individual young hens on this farm there was amplification and transmission of infection amongst members of the flock.  相似文献   

16.
A vaccine for necrotic enteritis (NE) of chickens would reduce the current need to prevent or treat the disease in broiler chickens with antimicrobial drugs. The objective of this study was to understand aspects of immunity to the disease. The first experiment examined the virulence of six strains of Clostridium perfringens isolated from cases of NE in broiler chickens. Using a 5-day experimental oral infection of 2-week-old broiler chickens, four of the six strains were found to be virulent. Pulsed-field gel electrophoresis and PCR showed that virulence was not associated with a plasmid encoding the beta2 toxin gene, cpb2, since this was present in virulent and one of the two avirulent strains. In the second experiment, two virulent and one avirulent strains were tested for their ability to immunize ("infection-immunization") chickens through the oral route. The procedure used experimental infection for 5 days followed by bacitracin treatment for 9 days, and then re-challenge 2 days later with a virulent strain, CP4. Infection-immunization with the virulent isolates protected chickens from subsequent virulent challenge, whereas the infection-immunization with the avirulent isolate did not. In a third experiment, two of four alpha-toxin-negative mutants of CP4 protected birds from experimental NE after oral immunization. These two mutants were also attenuated for virulence. We conclude that it is possible to immunize chickens successfully against NE and that immunogen(s) other than alpha-toxin are important in protective immunity against oral infection.  相似文献   

17.
In this study, we investigated risk factors associated with the probability to detect Salmonella in samples of litter collected within 2 h prior to new flock placement in 76 grow‐out houses on 38 conventional broiler farms located in the US states of Mississippi, Alabama and Texas. We evaluated characteristics of location and layout of the farm; area adjacent to and surrounding the house; house construction; condition and type of equipment in the house; litter management and other production, sanitation, visitation and biosecurity practices; non‐broiler animal species on the farm; and weather conditions on the 3 days leading up to flock placement. Logistic regression was used to model the relationships between probability to detect Salmonella in litter and potential risk factors. In the screening process, each risk factor was evaluated as a single fixed effects factor in a multilevel model that accounted for variability among the sampled farms and their production complexes and companies. Of almost 370 risk factors screened, 24 were associated with the probability to detect Salmonella in litter. These were characteristics of the surroundings of the house, house construction and conditions, litter management, length of downtimes between flocks in the house, biosecurity and farm location. After investigation of collinearity between these variables and building of models for important risk factor categories, the list of candidate variables for the final model was refined to eight factors. The final model demonstrated that a higher probability of detecting Salmonella in litter was strongly associated with the use of wood to construct the base of the walls or to cover the inside of the broiler house foundation, and with the use of fresh wood shavings to top‐dress or completely replace the litter between flocks.  相似文献   

18.
Anticoccidial vaccine and an anticoccidial drug rotation program were compared to determine which program was more effective in producing coccidia populations sensitive of 1 ppm diclazuril. The study used an anticoccidial drug-sensitivity battery test (AST) to determine the baseline level of diclazuril sensitivity to field isolates of Eimeria spp. from seven broiler complexes that had used diclazuril. Based on percentage reduction in weight gain and lesion scores, 25% or fewer of the isolates were effectively controlled by diclazuril. Following the baseline sampling, four of the complexes switched to a nondiclazuril in-feed anticoccidial drug program and three of the complexes switched to a vaccination program for two broiler grow-out cycles as the sole coccidiosis-control program. This study demonstrated that the vaccine used (Coccivac-B) contained anticoccidial drug-sensitive strains. Eimeria isolates were subsequently collected from the identical houses and diclazuril AST results were compared with the baseline AST results. Following the two grow-out cycles, sensitivity of the isolates to diclazuril from the four complexes that continued to use in-feed anticoccidial drugs remained essentially unchanged. The isolates from the three complexes that switched to the vaccination program demonstrated a marked increase in diclazuril sensitivity, with 60%-100% of the isolates from each complex effectively controlled by diclazuril. Vaccination with the anticoccidial drug-sensitive strains produced a measurable increase in the level of sensitivity to diclazuril.  相似文献   

19.
Due to the diminished use of growth-promoting antibiotics in the European Union, Clostridium perfringens induced necrotic enteritis and subclinical disease have become important threats to poultry health. A study was set up to genotypically and phenotypically characterise C. perfringens isolates from poultry flocks with different health status. Animals from healthy flocks were sampled by cloacal swabs, while intestinal and liver samples of animals suffering from necrotic enteritis were analysed. A total of 27 isolates was obtained from 23 broiler flocks without clinical problems and 36 isolates were obtained from 8 flocks with clinical problems. Using PFGE typing, high genetic diversity was detected between isolates from different flocks. Isolates derived from flocks where disease outbreaks occurred were clonal within each flock, but each flock harboured a different clone. All isolates were of toxin type A. Isolates from 5 out of 35 PFGE types carried the cpb2 gene, encoding the beta2 toxin, and isolates from 2 out of 35 PFGE types harboured the cpe gene, encoding the enterotoxin. In vitro alpha toxin production for all isolates was quantified by enzyme-linked immunosorbent assay. It was shown that in vitro alpha toxin production of C. perfringens isolates from diseased flocks was not higher than in vitro alpha toxin production from isolates derived from healthy flocks.  相似文献   

20.
This review presents a framework for assessing the efficacy of antimicrobials used to control necrotic enteritis (NE) caused byClostridium perfringens (CP) in the context of susceptibility testing and clinical efficacy, and their potential interactions with the intestinal microbiota of poultry. Practitioners have traditionally based their choice of antimicrobial agent on antimicrobial susceptibility testing, but there appears to be a lack of correlation with clinical efficacy for in-feed antimicrobials (particularly bacitracin and virginiamycin). Resistance patterns of CP and antimicrobials have been monitored using epidemiological cutoffs for minimal inhibitory concentration (MIC), which are not intended to guide therapy. Also, most data have been determined using CP isolates from healthy birds (i.e., potentially from commensal strains not known to be clinically relevant). It is believed that NE is caused by specific virulent CP strains (and potentially other bacteria) that proliferate and displace these commensals. The presence of resistant commensals is not necessarily detrimental (and may even be beneficial) if they inhibit the single CP strain dominance effect observed in acute NE. The choice of antimicrobial therapy in a clinical setting should thus be based on a variety of factors, including an accurate diagnosis, results of efficacy studies, prior experience at the premises in question, and interpretation of MIC data, recognizing that it is not necessarily well correlated with clinical efficacy.  相似文献   

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