Received 20 December 2007; returned 5 February 2008; revised 20 March 2008; accepted 4 April PDF Free Download

Received 20 December 2007; returned 5 February 2008; revised 20 March 2008; accepted 4 April 2008

Size: px
Start display at page:

Download "Received 20 December 2007; returned 5 February 2008; revised 20 March 2008; accepted 4 April 2008"

Transcription

1 Journal of Antimicrobial Chemotherapy (2008) 62, doi: /jac/dkn190 Advance Access publication 7 May 2008 Persistence of Campylobacter species, strain types, antibiotic resistance and mechanisms of tetracycline resistance in poultry flocks treated with chlortetracycline L. J. V. Piddock 1 *, D. Griggs 1, M. M. Johnson 1, V. Ricci 1, N. C. Elviss 2, L. K. Williams 2, F. Jørgensen 2, S. A. Chisholm 3, A. J. Lawson 3, C. Swift 3, T. J. Humphrey 4 and R. J. Owen 3 1 Antimicrobial Agents Research Group, Division of Immunity and Infection, The Medical School, University of Birmingham, Birmingham, UK; 2 Health Protection Agency Foodborne Zoonoses Unit, School of Clinical Veterinary Science, University of Bristol, Bristol, UK; 3 Centre for Infections, Health Protection Agency, London, UK; 4 Foodborne Zoonoses Group, School of Clinical Veterinary Science, University of Bristol, Bristol, UK Received 20 December 2007; returned 5 February 2008; revised 20 March 2008; accepted 4 April 2008 Objectives: The aim of this study was to investigate the persistence of Campylobacter species, strain types, antibiotic resistance and mechanisms of tetracycline resistance in poultry flocks treated with chlortetracycline. Methods: Three commercially reared broiler flocks, naturally colonized with Campylobacter, weretreated with chlortetracycline under experimental conditions. The numbers of Campylobacter isolated, and the species, flaa short variable region allele, and antimicrobial resistance of isolates were determined. Results: For two of three flocks, tetracycline-resistant strains predominated prior to chlortetracycline exposure. Presence of the antibiotic had no discernible effect on the numbers or types of Campylobacter and the tetracycline-resistant strains persisted in numbers similar to those observed before treatment. With all flocks, some faecal samples were obtained that contained no Campylobacter, irrespective of exposure to chlortetracycline; this was more common as the birds grew older. For the third flock, tetracycline-resistant Campylobacter were in the minority of samples before and during exposure to chlortetracycline, but at sampling times after this, no resistant strains were found in the treated (or untreated) birds, irrespective of exposure to the antibiotic. All tetracycline-resistant isolates (MICs 16 to >128 mg/l) contained tet(o) and, for some isolates, this was transferable to Campylobacter jejuni The efflux pump inhibitor PAbN reduced the MICs of tetracycline for these isolates by 4-fold, suggesting that an intact efflux pump, presumably CmeABC, is required for high-level tetracycline resistance. Conclusions: Our data indicate that chlortetracycline treatment does not eradicate tetracycline-resistant Campylobacter spp. from poultry. However, if a low number of resistant isolates are present, then the antibiotic pressure appears insufficient to select such strains as the dominant population. Keywords: chickens, typing, tetracycline-resistant Introduction Campylobacter jejuni causes the greatest number of food-borne bacterial infections in humans in England and Wales. 1 Infections by this pathogen also had the greatest impact on the healthcare sector in England and Wales between 1996 and 2000, giving rise to just under hospitalizations and 80 deaths. Adak et al. 1 showed that the most important cause of indigenous food-borne disease was contaminated chicken. Campylobacteriosis is usually self-limiting but antibiotic treatment is required for chronic and serious infections. Resistance to antibiotics has become a serious problem worldwide, and the numbers of Campylobacter resistant to multiple antibiotics continue to increase. Antibiotics are administered to treat infections in animals caused by other bacteria, and antibiotic-resistant Campylobacter can emerge as a consequence of this, contaminate food and enter the human food chain. Chlortetracycline is a broad-spectrum antibacterial agent, with activity against respiratory and enteric bacteria, and is the most... *Corresponding author. Tel: þ ; Fax: þ ; Present address. Health Protection Agency Yorkshire and the Humber, Leeds, UK # The Author Published by Oxford University Press on behalf of the British Society for Antimicrobial Chemotherapy. All rights reserved. For Permissions, please

2 Piddock et al. commonly used therapeutic antibiotic in poultry production. In 2004, 243 tonnes of antimicrobials belonging to the tetracycline class were sold in the UK. 2 Tetracycline-resistant C. jejuni isolated from chickens have been described by various authors since Taylor et al. 3 The rate of resistance in live chickens and meat at retail sale varies from country to country. Various studies have shown that depending on the origin and type of poultry meat, between 3% and 35% of the Campylobacter are resistant to tetracycline. 4 7 Of interest, Anderson et al. 4 have shown that there has been an increase in the numbers of resistant isolates in Denmark since 2001, which coincides with the withdrawal of antimicrobial growth promoters in Europe. In the USA and Canada, Campylobacter were highly prevalent in both conventional and organic poultry production, and large numbers of isolates were resistant to tetracycline. 8,9 Approximately 25% of the Campylobacter isolates from humans have also been reported as tetracycline-resistant. Most recently, 27.3% of the C. jejuni isolated from patients in England and Wales with non-travelassociated cases of enteritis were resistant to tetracycline compared with 55% of the C. jejuni isolated from travel-associated cases (Iain Gillespie, HPA, Colindale, UK, personal communication). Taylor et al. 3 were the first to show that poultry isolates of C. jejuni contained a conjugative plasmid that transferred tetracycline resistance. Numerous studies have since shown that tetracycline resistance is typically transferable, and a variety of plasmids have been implicated. Manavathu et al. 10 found that the gene transferred was tet(o). Avrain et al. 11 showed that there was horizontal transfer of tet(o)-positive Campylobacter between chickens. However, tet(o) can also be located on the chromosomes Decreased susceptibility to tetracycline can also be mediated by the CmeABC efflux pump along with resistance to several other drugs. 15,16 It has been suggested that in the absence of an antimicrobial agent, antibiotic-resistant bacteria are less prevalent. Therefore, it has been postulated that in the presence of a drug (e.g. tetracycline), drug-resistant bacteria would be maintained. In our previous work on fluoroquinolone resistance in C. jejuni, we monitored the persistence of resistant strains before, during and after poultry flocks were exposed to a fluoroquinolone and showed a clear association between exposure to fluoroquinolones and the presence of resistant strains which persisted in the birds to slaughter and entry into the human food chain. 17,18 Based on our previous work, we hypothesized that exposure to any antibiotic used to treat infections in poultry would allow resistant Campylobacter to dominate the population. In this study, we investigated the persistence of Campylobacter species, strain types, antibiotic resistance and mechanisms of tetracycline resistance in poultry flocks treated with chlortetracycline. Materials and methods Treatment and sampling of chicken flocks All three flocks, A, B and C, were broiler birds removed from commercial poultry flocks reared for human consumption. The treatment histories of the birds were obtained and none had previous exposure to tetracycline or other antimicrobials. These birds were housed at the School of Clinical Veterinary Science of the University of Bristol, at a stocking density of 12.6 kg/m 2 to meet the UK Home Office requirements and experimentally treated with chlortetracycline. All animal experiments were conducted according to the requirements of the Animals (Scientific Procedures) Act 1986 and were approved by the local Ethics Review Committee. Each group of birds was housed independently in separate rooms. The University of Bristol s Animal Services Units biosecurity protocol was followed. This included the use of protective clothing (overalls, gloves and disposable boot socks), which was worn throughout husbandry and sampling. Separate protective clothing and footwear was worn for each room, and disinfectant boot dip used upon every entry and exit of the rooms. The footwear worn was dedicated to the animal house and had not previously been worn outside. Flock A consisted of 20 birds brought on-site at 21 days of age from a commercial free-range organic broiler flock consisting of 7500 birds in September All birds were treated with a therapeutic dose of chlortetracycline at 40 mg/kg/day for 6 days administered in the feed as recommended by the manufacturer (Aurogran; Novartis, Hertfordshire, UK). All birds were weighed before therapy commenced. Food consumption was not measured; however, animal care staff confirmed that the animals fed normally. Flock B consisted of 28 birds brought on-site in October 2005, purchased from a commercial free-range broiler flock of 6300 birds at 22 days old. Fourteen of these birds were treated with a therapeutic dose of chlortetracycline and 14 birds, housed independently, remained untreated. Flock C consisted of 30 birds brought on-site at 49 days old in May 2006, purchased from a commercial housed corn-fed freedom foods flock consisting of birds. Fifteen of these birds were treated with a therapeutic dose of chlortetracycline and 15 birds, housed independently, remained untreated. All samples were transported to the laboratory within 3 h of collection. At least 14 freshly voided faecal samples were collected before, during (2 days after treatment started) and every 7 days thereafter up to 4 weeks post-treatment, and Campylobacter spp. were isolated on modified charcoal cefoperazone deoxycholate agar (mccda; Oxoid, Basingstoke, UK) as described previously. 17 At 4 weeks post-treatment sampling, the caeca from flock B were removed post mortem to enable analysis of each chicken s flora for Campylobacter spp. Enumeration of tetracycline-resistant Campylobacter The proportion of tetracycline-resistant Campylobacter was determined in the faecal sample isolates by replica-plating (using sterile furniture grade velvet) colonies from the master plate of bacteria growing on mccda onto Mueller Hinton agar (Oxoid), containing CCDA selective supplement (Oxoid), 5% defibrinated horse blood (Oxoid) and 8 mg/l tetracycline (Sigma, Dorset, UK). This cut-off concentration of 8 mg/l tetracycline was used as a review of the literature had indicated that this differentiated tetracycline-resistant strains from those that were tetracycline-susceptible. Replica plates were incubated at 378C for 48 h in a microaerobic atmosphere (5% to 6% O 2,3%to7%CO 2 and 7% H 2, in a balance of nitrogen 19 ). The number of colonies growing on the replica plate was determined, and their morphology compared with growth on the master mccda plates from which they were produced and the proportions of resistant colonies in the faecal samples were determined by subtraction. Usually, the lowest dilution was replica plated, but for some samples from flock C, this was not possible due to overgrowth of other faecal flora; hence, the different detection limits for the resistant population are indicated by horizontal lines on the bars in Figure 3 for this flock. 304

3 Antibiotic resistance in chlortetracycline-treated poultry flocks Identification of presumptive colonies as Campylobacter spp. Presumptive Campylobacter colonies (up to three colonies per sample) were subcultured from mccda to Columbia blood agar containing 5% defibrinated horse blood (Oxoid) and incubated at 378C for 48 h in a microaerobic atmosphere. Isolates were confirmed as Campylobacter using light microscopy for motility and cell morphology and lack of growth in air at 378C after 48 h. Species identification Speciation of C. jejuni and Campylobacter coli was performed using the method of Best et al., 20 employing a real-time PCR assay based on the ABI PRISM 7700 Sequence Detection System (Taqman) platform. Isolates that were not identified as Campylobacter spp. were subcultured under aerobic conditions, and any isolates that were aerotolerant were speciated as Arcobacter butzleri, Arcobacter cryaerophilus or Arcobacter skirrowii, according to the multiplex PCR assay of Houf et al. 21 Flagellin gene (flaa) short variable region (SVR) sequence typing Genotyping based on flaa SVR type was performed as follows. DNA templates were generated from chromosomal DNA recovered from boiled whole-cell suspensions using PCR primers and protocols described by Nachamkin et al. 22 Template preparations were purified using the Whatman w 96 Well PCR Cleanup Kit, and quantification of the products was performed by electrophoresis on a 1% (w/v) agarose gel. Sequencing primers and protocols for the SVR analysis were carried out as described by Meinersmann et al. 23 Sequencing was performed using a Beckman CEQ800 Genetic Analysis System, and also commercially (K-Biosciences, Hoddesdon, UK). Forward and reverse sequences were aligned and trimmed to a 321 bp region covering the flaa SVR of genomesequenced strain C. jejuni NCTC using Bionumerics V2.0 software (Applied Maths, Kortrijk, Belgium). The 321 bp sequences were then compared with flaa sequences (870 alleles at the time of writing) held in an online database ( uk/flaa/), and the corresponding allele numbers were recorded. The flaa SVR types were designated based on 100% homology to the Oxford reference sequences. PFGE and profile analysis PFGE was performed according to the protocol of Gibson et al., 24 when it was considered necessary to check any anomalous results. DNA was digested with SmaI. For example, when isolates had the same flaa SVR type but different antibiotic susceptibility patterns (resistance phenotype) or when an isolate was non-typeable by flaa SVR. PFGE gel profiles were arbitrarily assigned numbers by comparison (visually and by using BioNumerics V2.0) with profiles of other isolates tested as part of a larger collaborative study examining additional flocks (data not shown). Determination of antimicrobial resistance Initially, all isolates were screened for antibiotic susceptibility (no growth) or resistance (growth) by the breakpoint screening method of Thwaites and Frost, 25 with minor modifications such that the conditions were the same as those used to determine the MIC values (see below). The following concentrations of antimicrobials were used: ampicillin, 8 and 32 mg/l; chloramphenicol, 8 mg/l; gentamicin, 4 mg/l; kanamycin, 16 mg/l; neomycin, 8 mg/l; tetracycline, 8 and 128 mg/l; nalidixic acid, 16 mg/l; ciprofloxacin, 1 mg/l; and erythromycin, 4 mg/l. Isolates were selected for further study on the basis of species, flaa SVR type and resistance phenotype determined by breakpoint screening. MICs were determined for every strain of each flaa SVR and PFGE type from each and every Campylobacter-positive sample from each treatment phase of all flocks. Where there was more than one phenotype (by flaa SVR, PFGE or breakpoint screening) present within a sample, each phenotype was examined. Bacteria were grown on Mueller Hinton agar containing 5% horse blood at 368C in 7.5% CO 2. The agar doubling dilution procedure recommended by the CLSI (formerly the NCCLS) Campylobacter Working Group 26 was used throughout the study, as described previously, 13 to determine the MICs of selected antibacterial agents and dyes (chlortetracycline, tetracycline, ampicillin, amoxicillin, ciprofloxacin, nalidixic acid, erythromycin, chloramphenicol, kanamycin, lincospectin, triclosan and ethidium bromide). C. jejuni NCTC and C. coli NCTC were used as control strains. Designation of strains as antibiotic-susceptible or -resistant was made as described previously, 18 with reference to the guidelines of the BSAC and CLSI as available at the start of this study in PCR detection of tet(o) and transfer of tetracycline resistance All isolates inhibited by 16 mg/l tetracycline were examined for the presence of tet(o) in cell lysates using primers based on the sequence published by Manavathu et al. 10 and amplified from nucleotides Furthermore, 13 isolates representative of resistant strains isolated from each flock were examined for their ability to transfer tetracycline resistance to C. jejuni as described by Taylor et al. 3 These were examined in parallel to 11 tetracycline-resistant isolates from our previous study. 17,18 The DNA sequence of tet(o) of four isolates from our previous studies and three isolates from flock A was sequenced. These represented isolates with a range of MIC values from 16 to.128 mg/l tetracycline and examples of transferable and non-transferable tet(o). Results Effect of treatment with chlortetracycline upon numbers of Campylobacter isolated During the course of this study, we were alerted to three commercial poultry flocks being reared for human consumption that were naturally infected with Campylobacter. Birds were purchased from these flocks and experimentally treated with a therapeutic dose of chlortetracycline exactly as if treated in the commercial environment. For flock A, data were only obtained for the birds treated with chlortetracycline (Figure 1). For flocks B (Figure 2) and C (Figure 3), sufficient birds were obtained to allow them to be divided into two groups: one group remained untreated and one was treated with chlortetracycline. The treated and untreated groups were housed independently. Pre-treatment, 19/20 faecal samples from flock A contained tetracycline-resistant Campylobacter. Except for one sample where a small proportion of the colonies were susceptible to tetracycline, replica plating showed that in all samples, every colony was resistant to tetracycline (Figure 1 and Table 1). During treatment, all samples contained Campylobacter spp. but the number of Campylobacter isolated from each varied, and some samples contained tetracycline-susceptible isolates (indicated by white bars in Figure 1). Faecal samples were obtained up to 4 305

4 Piddock et al. Figure 1. Numbers of Campylobacter spp. in chicken faeces collected from flock A before, during and after chlortetracycline (CTC) treatment. Each bar represents an individual freshly voided faecal sample. Black bars indicate the numbers of tetracycline-resistant Campylobacter spp., whereas the white bars indicate the proportion of tetracycline-susceptible Campylobacter spp. Grey bars indicate samples for which no Campylobacter were detected. The y-axis is the number (log 10 )ofcampylobacter spp. per gram of chicken faeces. The x-axis shows the sampling time; 20 samples were taken at each sampling point. weeks post-treatment, during which time the number of samples containing Campylobacter reduced, although tetracycline-resistant strains were isolated up to slaughter. For example, 2 weeks posttreatment in 3/20 samples, no Campylobacter were detected (indicated by grey bars in the figures), and by 4 weeks posttreatment, 8/20 samples contained no detectable Campylobacter. Tetracycline-resistant strains were isolated from most samples from flock B pre-treatment and during treatment, irrespective of whether the birds had been exposed to chlortetracycline (Figure 2 and Table 1). Unfortunately, despite repeated attempts with the raw material, due to overgrowth of contaminants on the replica plates 1 week post-chlortetracycline treatment, the proportion of samples from the treated birds containing tetracycline-resistant Campylobacter could not be calculated with accuracy (indicated by hatched bars in Figure 2). However, all samples from the untreated flock taken at the same time period contained tetracycline-resistant Campylobacter. Two weeks after treatment with chlortetracycline and up to slaughter, samples containing Campylobacter were obtained from both the treated and untreated birds, and all these contained tetracycline-resistant isolates. As with flock A, in the latter stages of sampling (2 4 weeks posttherapy), some samples from both untreated and treated birds contained no Campylobacter (5/42 and 8/41 samples, respectively). Birds from the source flock were also sampled at the original farm immediately prior to slaughter; all 14 samples contained Campylobacter, of which 2 contained tetracycline-resistant Campylobacter. Flock C contained both tetracycline-susceptible and -resistant Campylobacter (Figure 1 and Table 1) but in some samples from this flock no Campylobacter were detected. There was a different detection limit for resistant Campylobacter in some samples as overgrowth of contaminating faecal flora prevented replica plating of the lower dilutions. Prior to treatment, 2/15, and 2/15, faecal samples from untreated and treated birds, respectively, contained tetracycline-resistant Campylobacter. During treatment, 4/10 samples from the untreated birds and 4/8 from treated birds contained tetracycline-resistant Campylobacter. Following treatment, all samples contained Campylobacter, of which only one contained tetracycline-resistant Campylobacter; thereafter, no tetracycline-resistant Campylobacter were isolated. Over the same time period, no tetracycline-resistant Campylobacter were isolated from the untreated birds. As with flocks A and B, there was a decline in the number of samples from the birds (treated or untreated) from which Campylobacter were detected. Despite the decline, greater numbers of Campylobacter were obtained from the chlortetracycline-treated birds than the untreated birds. Effect of treatment with chlortetracycline upon species, type and antimicrobial susceptibility of Campylobacter isolated Up to three colonies from each of the 14 faecal samples per sampling time were flaa SVR genotyped. PFGE was also performed on representative isolates of each flaa type from each sampling period, from treated and untreated birds, and for those isolates for which no flaa type could be obtained. The PFGE patterns were consistently associated with the same flaa type, so that for those isolates for which a flaa type could not be obtained, the PFGE pattern indicated the strain type, which was further confirmed by MIC testing. Samples from birds in flock A contained two flaa SVR types, 16 and 18, that were previously recorded by others in the Oxford database. flaa18 was further divided into P3a and P3b types by PFGE. These PFGE types differed by the size of a single band only, and both PFGE types had the same phenotype upon MIC testing. The majority of the isolates from the faecal samples from flock A were flaa18. Not only were there fewer isolates of C. jejuni flaa16, but this strain was not detected from faecal samples taken at 2, 3 and 4 weeks after treatment. On initial breakpoint testing, three phenotypes were identified (Table 1). However, the determined MIC values of 306

5 Antibiotic resistance in chlortetracycline-treated poultry flocks Figure 2. Numbers of Campylobacter spp. in chicken faeces collected from flock B before, during and after chlortetracycline (CTC) treatment. Each bar is as described for Figure 1. The hatched bars indicate those samples in which the numbers of tetracycline-resistant bacteria could not be estimated due to overgrowth of contaminants. The y-axis is the number (log 10 )ofcampylobacter spp. per gram of chicken faeces. The x-axis shows the sampling time; 14 samples were taken at each sampling point. the 12 agents indicated that they were indistinguishable from each other (Table 2). All flaa18 isolates, irrespective of when isolated from flock A, were resistant to chlortetracycline, tetracycline, ciprofloxacin, nalidixic acid and triclosan. Interestingly, the C. jejuni flaa18 isolated became progressively less susceptible to triclosan as sampling progressed (Table 2). The MIC values were typically 16 mg/l triclosan pre-treatment and by 2 weeks post-treatment the MICs for isolates of this strain were consistently 64 mg/l. This decline in susceptibility was not observed with any other agent or with the isolates from flocks B and C. Samples from treated and untreated birds in flock B contained two species of Campylobacter, C. jejuni and C. coli, each with previously recorded flaa SVR types flaa21 and flaa66, respectively. Each type was associated with unique PFGE patterns P22 and P23, respectively (Table 2). As with the strains from flocks A and B, these flaa types had been found previously by others. Before treatment, no C. coli flaa66 were isolated from faeces from the untreated birds, but it was recovered from the treated birds. However, this strain was isolated at all other times from both groups of birds. Consistently, at each sampling time, greater numbers of C. jejuni flaa21 were isolated than C. coli flaa66. All C. jejuni flaa21 were resistant to chlortetracycline, tetracycline, amoxicillin and triclosan. All isolates of C. coli flaa66 were resistant to the same agents plus nalidixic acid. Samples taken from the source flock 4 weeks after treatment also contained Campylobacter; all were C. jejuni flaa14 PFGE 8a (Table 2). Five isolates were resistant to triclosan only and one isolate was resistant to chlortetracycline, tetracycline, nalidixic acid, amoxicillin and triclosan. 307

6 Piddock et al. Figure 3. Numbers of Campylobacter spp. in chicken faeces collected from flock C before, during and after chlortetracycline (CTC) treatment. Each bar is as described for Figure 1. For this flock, the detection limit for the numbers of tetracycline-resistant Campylobacter is illustrated as a horizontal line as it was not always possible to replica plate the lowest dilution of sample due to overgrowth of contaminants. The y-axis is the number (log 10 )ofcampylobacter spp. per gram of chicken faeces. The x-axis shows the sampling time; 15 samples were taken at each sampling point. Samples from treated and untreated birds in flock C contained two species of Campylobacter, C. jejuni and C. coli (flaa255). Three strains of C. jejuni (flaa85, flaa189 and flaa239) were isolated, each associated with unique PFGE patterns; P24a, P25 and P26, respectively (Table 2). Except for one sampling time (untreated birds at the pre-treatment sampling point), C. coli flaa255 was isolated at all times from both groups of birds. Overall, for both groups, higher numbers of C. coli flaa255 were isolated. Of the C. jejuni, flaa85 was the most commonly isolated strain of this species. Except for triclosan, all isolates of C. coli flaa255, C. jejuni flaa189 and flaa239 (also resistant to amoxicillin) were susceptible to all other agents tested. Two isolates of C. jejuni flaa85 and one of C. coli flaa255 from the treated birds faecal samples had a markedly different phenotype upon MIC testing compared with the other isolates of the same flaa type. These isolates were less susceptible to chlortetracycline, tetracycline and ciprofloxacin or chloramphenicol. Mechanisms and transfer of tetracycline resistance Of the tetracycline-resistant isolates (MIC 16 mg/l) examined by PCR for the presence of tet(o), all were found to contain the gene (Table 2). Those isolates not tested were of the same genotype and phenotype obtained from the same sample and considered likely to be identical. Tetracycline-susceptible isolates of each type and from the same sampling time did not contain tet(o). Eight of 13 isolates from the present study and 6/11 isolates from our previous study 13 were able to transfer tetracycline resistance to C. jejuni at a frequency transfer of The MIC of tetracycline for three transconjugants was reproducibly 308

7 Table 1. Sampling, speciation and typing of Campylobacter spp. isolated from chicken flocks before, during and post-treatment with chlortetracycline (CTC) Flock Treatment phase No. of Campylobacter-positive sample (no. of samples) Total no. of isolates typed (total no. of colonies) No. of isolates of each type a Species flaa SVR type Resistotype PFGE type (no. tested) No. of isolates for which susceptibility was tested a 309 Flock A CTC-treated pre 19 (20) 56 (57) 56 C. jejuni 18 AMP-TET-NAL-CIP ND 19 during 20 (20) 60 (60) 54 C. jejuni 18 AMP-TET-NAL-CIP ND 17 3 C. jejuni 18 AMP P3a (3) 1 3 C. jejuni 16 AMP P4 (3) 1 1 week 20 (20) 60 (60) 51 C. jejuni 18 AMP-TET-NAL-CIP P3a (1) 18 3 C. jejuni 16 AMP P4 (3) 2 4 C. jejuni 16 AMP-TET-NAL-CIP P4 (2) 1 2 C. jejuni NR AMP-TET-NAL-CIP ND 1 2 weeks 17 (20) 51 (51) 28 C. jejuni 18 AMP-TET-NAL-CIP P3a (10) C. jejuni 18 TET-NAL-CIP P3a (10), P3b (1) 11 3 weeks 16 (20) 48 (48) 30 C. jejuni 18 AMP-TET-NAL-CIP P3a (8), P3b (2) C. jejuni 18 TET-NAL-CIP P3a (6), P3b (2) 11 4 weeks 12 (20) 36 (36) 36 C. jejuni 18 TET-NAL-CIP ND 12 Flock B untreated pre 24 (28) 67 (67) 67 C. jejuni 21 AMP-TET P22 (3) 14 during 14 (14) 42 (42) 41 C. jejuni 21 AMP-TET ND 5 1 C. jejuni NR AMP-TET P22 (1) 0 1 week 14 (14) 42 (42) 33 C. jejuni 21 AMP-TET ND 3 9 C. coli 66 AMP-TET P23 (2) 3 2 weeks 13 (14) 38 (39) 36 C. jejuni 21 AMP-TET ND 5 2 C. coli 66 AMP-TET ND 1 3 weeks 13 (14) 36 (39) 35 C. jejuni 21 AMP-TET ND 5 1 C. jejuni NR AMP-TET P22 (1) 0 4 weeks 11 (14) 33 (33) 32 C. coli 66 AMP-TET ND 5 1 C. coli NR AMP-TET P23 (1) 0 Birds at farm before 14 (14) 40 (42) 38 C. jejuni 14 AMP P8a (3) 7 slaughter 2 C. jejuni NR AMP ND 0 Flock B CTC-treated during 14 (14) 42 (42) 39 C. jejuni 21 AMP-TET P22 (2) 7 2 C. jejuni NR AMP-TET P22 (2) 2 1 C. coli 66 AMP-TET ND 1 1 week 14 (14) 40 (40) 38 C. jejuni 21 AMP-TET P22 (1) 14 2 C. coli 66 AMP-TET ND 1 2 weeks 13 (14) 39 (39) 36 C. jejuni 21 AMP-TET ND 11 3 C. coli 66 AMP-TET P23 (2) 1 Continued Antibiotic resistance in chlortetracycline-treated poultry flocks Downloaded from at Pennsylvania State University on February 27, 2014

8 Table 1. Continued Flock Treatment phase No. of Campylobacter-positive sample (no. of samples) Total no. of isolates typed (total no. of colonies) No. of isolates of each type a Species flaa SVR type Resistotype PFGE type (no. tested) No. of isolates for which susceptibility was tested a weeks 11 (14) 21 (21) 12 C. jejuni 21 AMP-TET ND 5 9 C. coli 66 AMP-TET P23 (2) 3 4 weeks 11 (13) 33 (33) 32 C. coli 66 AMP-TET P23 (1) 11 1 C. coli NR AMP-TET ND 1 Flock C untreated pre 7 (15) 13 (15) 6 C. jejuni 189 ND 3 3 C. jejuni 239 P26 (1) 3 2 C. jejuni 85 ND 2 2 C. jejuni ND P24a (1) 2 during 10 (15) 28 (30) 5 C. jejuni 189 P25 (2) 3 12 C. jejuni 85 P24a (2) 6 4 C. jejuni ND P24a (4) 1 7 C. coli 255 AMP P7b (1) 4 1 week 12 (15) 32 (36) 2 C. jejuni 189 ND 1 1 C. jejuni 239 ND 1 4 C. jejuni 85 P24a (1) 2 25 C. coli 255 AMP ND 10 2 weeks 7 (15) 17 (21) 3 C. jejuni 189 P25 (1) 1 1 C. jejuni 239 ND 1 1 C. jejuni 85 ND 1 12 C. coli 255 AMP ND 4 3 weeks (faeces) 5 (15) 13 (15) 3 C. jejuni 239 ND 1 3 C. jejuni ND P24a (3) 1 7 C. coli 255 AMP ND 3 2 (2) 6 (6) 1 C. jejuni 85 ND 1 Piddock et al. Downloaded from at Pennsylvania State University on February 27, 2014

9 311 3 weeks (caeca) 5 C. coli 255 AMP ND 2 Flock C CTC-treated during 8 (15) 20 (21) 6 C. jejuni 189 ND 3 3 C. jejuni 239 P26 (1) 1 6 C. jejuni 85 P24a (3) 4 2 C. jejuni ND P24a (2) 1 3 C. coli 255 AMP ND 2 1 week 13 (15) 35 (35) 7 C. jejuni 189 ND 4 1 C. jejuni 239 ND 1 9 C. jejuni 85 P24a (2) 6 3 C. jejuni ND P24a (3) 1 15 C. coli 255 AMP ND 6 2 weeks 6 (15) 13 (13) 4 C. jejuni 189 P25 (2) 2 9 C. coli 255 AMP P7b (1) 4 3 weeks 9 (15) 23 (23) 4 C. jejuni 189 ND 2 3 C. jejuni 85 P24a (2) 2 3 C. jejuni ND P24a (1), P24b (1), P25 (1) 2 3 weeks (caeca) 13 C. coli 255 AMP ND 5 2 (2) 6 (6) 3 C. jejuni 189 P25 (2) 1 2 C. coli 255 AMP ND 1 1 C. coli 255 AMP-NAL-CIP P7b (1) 1 AMP, ampicillin; TET, tetracycline; CIP, ciprofloxacin; NAL, nalidixic acid; ND, not done; NR, flaa SVR typing failed to give data on several attempts;, fully antibiotic susceptible. a MICs were determined for every strain of each flaa SVR and PFGE type from each and every Campylobacter-positive sample from each treatment phase of all flocks. Where there was more than one phenotype (by flaa SVR, PFGE or breakpoint screening) present within a sample, each phenotype was examined. The concordance between breakpoint testing (Table 1) and MIC determination (Table 2) was generally good. However, some differences arose. These were when the MIC value was +1 dilution of the cut-off concentration used in breakpoint testing. Thus, an isolate could fail to grow on the breakpoint plate (8 mg/l) but the MIC value was 16 mg/l. Antibiotic resistance in chlortetracycline-treated poultry flocks Downloaded from at Pennsylvania State University on February 27, 2014

10 Table 2. Susceptibility, typing and presence of tet(o) of Campylobacter isolated from the flocks before, during and post-therapy with chlortetracycline MIC (mg/l) Species flaa SVR type PFGE n CTC TET CIP NAL ERY AMX CHL Tric EtBr tet(o)-positive 312 CTC-treated flock A C. jejuni flaa18 3a, 3b a 117 range to b mode c C. jejuni flaa mode d Untreated flock B C. jejuni flaa range to ND b mode C. coli flaa mode b CTC-treated flock B C. jejuni flaa range to to b mode C. coli flaa range to to b mode Untreated birds at farm (prior to slaughter) C. jejuni flaa14 e 8a 6 mode d C. jejuni flaa14 e 8a Untreated flock C C. jejuni flaa85 24a 16 range d mode C. jejuni flaa mode d C. jejuni flaa mode d C. coli flaa255 P7b 23 range d mode CTC-treated flock C C. jejuni flaa85 24a 13 range d mode C. jejuni flaa85 24a 2 mode C. jejuni flaa range d mode C. jejuni flaa d C. jejuni flaa d C. coli flaa255 P7b 20 range d mode C. coli flaa255 P7b Piddock et al. CTC, chlortetracycline; TET, tetracycline; CIP, ciprofloxacin; NAL, nalidixic acid; ERY; erythromycin; AMX, amoxicillin; CHL, chloramphenicol; EtBr, ethidium bromide; Tric, triclosan; ND, not done. Where no range has been indicated the values shown represent the majority or all isolates of that phenotype. a PFGE indicated that there were two strains of an flaa type but the MICs for both strains were identical. b All Tet R isolates tested were positive for tet(o). c mode is the MIC value most frequently observed. d Only CTC/TET R isolates tested. e flaa14 P8a only isolated from birds sampled from the supplying farm at slaughter. Downloaded from at Pennsylvania State University on February 27, 2014

11 Antibiotic resistance in chlortetracycline-treated poultry flocks higher than that for the original isolate. The DNA sequences of tet(o) were also examined for seven representative isolates to determine whether there was any sequence variation associated with a particular MIC of tetracycline. The most resistant isolates (MIC 128 mg/l) contained an adenine at nucleotides 884, 1036 and 1784 of tet(o) and thymidine at nucleotide This is identical to the tet(o) sequence deposited in GenBank (accession no. M18896). Other sequence differences that gave amino acid substitutions were found (nucleotides 680, 910, 993 and 1063), but none could be correlated with tetracycline MIC values. In the present study, all tetracycline-resistant strains contained tet(o), but in a previous study, 18 there were eight tetracycline-resistant isolates for which tet(o) was not detectable by PCR. Likewise in studies on the effect of amoxicillin and tylosin performed in parallel to the present study, there were nine tetracycline-resistant isolates for which tet(o) was not detectable by PCR. To determine the role of efflux, and as variable MICs of ethidium bromide had been obtained (suggesting active efflux), in this phenotype MICs of tetracycline were determined in the presence and absence of the efflux pump inhibitor PAbN. For comparison, the effect upon susceptibility of PAbN (20 mg/l) was also examined for one tetracycline-resistant isolate of each flaa type from the present study. The MICs of the isolates for which tet(o) was not detectable by PCR ranged from 16 to 128 mg/l, but in the presence of PAbN, all values were reduced by 4- to 16-fold. PAbN had no effect upon the tetracycline susceptibility of C. jejuni cmeb::aph, but for a cmeb-overexpressing mutant, P1048, 16 the MIC was reduced from 8 to 0.12 mg/l. For those isolates containing tet(o), the MICs of tetracycline were reduced 4-fold in the presence of PAbN. Discussion In this study, we examined the effect of chlortetracycline treatment upon the numbers of Campylobacter isolated, their species, type and antimicrobial resistance from three naturally colonized flocks. The flocks were housed at commercially relevant stocking densities and treated exactly as in the commercial environment. Therefore, this study is the first to replicate the on-farm situation. For each, the source flock was reared commercially for entry into the human food chain. For flocks A and B, tetracycline-resistant strains predominated prior to chlortetracycline exposure and perhaps, not surprisingly, the presence of the antibiotic had no discernible effect on the numbers of Campylobacter and tetracycline-resistant strains persisted. However, for all three flocks, some faecal samples were obtained that contained no Campylobacter, irrespective of exposure to chlortetracycline; this was more common as the birds grew older. For flock C, low-level tetracycline-resistant Campylobacter were detected in a few samples before and during exposure to chlortetracycline, but at sampling times after this, no resistant strains were found in the treated birds, irrespective of exposure to the antibiotic. These data suggest that for these tetracycline-resistant Campylobacter, the resistance was not sufficient to allow the strain to multiply in the presence of the drug. This is opposite to the situation found previously with fluoroquinolones where the antibiotic pressure allowed the resistant strains to proliferate and predominate. 17 By performing genotyping, it was possible to determine whether tetracycline-resistant strains emerged during antibiotic exposure or if original resistant strains were selected and became the dominant population during therapy. It was also possible to establish if these persisted once the antibiotic had been withdrawn. Data obtained indicate that the original tetracycline-resistant strains persisted in flocks A and B. Only for flock C were Campylobacter of the same strain type but with different phenotypes (antibiotic susceptibilities) isolated, and these were only obtained from samples from the treated birds. However, only low numbers of isolates of each phenotype were obtained, so it is not possible to state whether these were present but not detected prior to antibiotic exposure or they emerged as a consequence of chlortetracycline treatment. Most of the tetracycline-resistant strains were cross-resistant to at least one other class of antibiotics, either a quinolone or b-lactam. No multidrug-resistant isolates (i.e. resistant to three or more classes of antimicrobial agent) were obtained. Although there is no recommended breakpoint concentration for triclosan, all isolates from the three test flocks required at least 32 mg/l triclosan for inhibition. Therefore, these isolates were compared with those from our previous study on the effect of fluoroquinolone treatment upon Campylobacter isolated from poultry flocks. 18 In that study, the MIC of triclosan for all multidrug-resistant isolates (n ¼ 97) was determined and a range of mg/l was identified; with an MIC at which 50% of the isolates were inhibited (MIC 50 ; median), MIC 90, and mode MIC of 64 mg/l. Taken together, these data indicate that irrespective of any antibiotic resistance, Campylobacter spp. are relatively insensitive to this biocide when compared with other Gram-negative bacteria. This may have implications for the eradication of food-borne Campylobacter from the domestic and industrial kitchen, where triclosan is commonly used. As has been found in previous studies, the presence of tet(o) in the current study was always associated with tetracycline resistance and could be transferred to C. jejuni The fact that not all tetracycline resistance was transferable suggests that for some isolates, tet(o) is chromosomally encoded, similar to the findings of others. 13,14,27 The MIC of tetracycline for some transconjugants was reproducibly higher than that of the original isolate, suggesting that the host strain can influence the level of resistance expressed. Unlike Gibreel and Taylor, 27 we did not find any variant tet(o) sequences associated with high-level resistance, because no isolates required 512 mg/l tetracycline for inhibition. The present study gave different conclusions to those from a previous study investigating the effect of tetracycline antibiotic (oxytetracycline) exposure upon Campylobacter in poultry flocks. 12 However, in their study, the single flock examined had no tetracycline-resistant strains before, during or after antibiotic exposure. As none of the Campylobacter became tetracyclineresistant during or after antibiotic exposure, it was concluded that no strains acquired a resistance gene such as tet(o) from the commensal enterococcal flora that were present concurrently. 12 Although in the current study all tetracycline-resistant strains contained tet(o), previous and parallel studies by ourselves with other antibiotics also provided strains that were tetracycline-resistant that were determined as tet(o)-positive by PCR. The presence of the efflux pump inhibitor PAbN reproducibly reduced the MICs of all strains tested, irrespective of the presence of tet(o). These data suggest that overexpression of an 313

12 Piddock et al. efflux pump can confer tetracycline resistance in the absence of tet(o) and that an intact efflux pump, presumably CmeB, is required for high-level tetracycline resistance even when the tet(o) gene is present. These data are similar to those of previous studies that indicated that CmeB was required for macrolide resistance in Campylobacter 28 and for various plasmid and chromosomally mediated antibiotic resistances in clinical isolates of Salmonella. 29 In summary, our data indicate that chlortetracycline treatment does not eradicate tetracycline-resistant Campylobacter spp. from poultry. However, if a low number of resistant isolates are present, then the antibiotic pressure appears insufficient to select such strains as the dominant population. Further studies to determine the Campylobacter population dynamics in flocks containing birds colonized with low numbers of antibiotic-resistant Campylobacter are now required. Acknowledgements We thank Sangita Bhattarai Sapkota, Zaahira Gani and Saba Ghori for providing technical support. Funding This work was supported by Defra project VM2200. No other support for this work was received. Transparency declarations None to declare. References 1. Adak GK, Meakins SM, Yip H et al. Disease risks from foods, England and Wales, Emerg Infect Dis 2005; 11: Anonymous. Veterinary Medicines Directorate. gov.uk/publications/antibiotic/antimicrob pdf (20 December 2007 date last accessed). 3. Taylor DE, de Grandis SA, Karmali MA et al. Transmissible plasmids from Campylobacter jejuni. Antimicrob Agents Chemother 1981; 19: Anderson SR, Saadbye P, Shukri NM et al. Antimicrobial resistance among Campylobacter jejuni isolated from raw poultry meat at retail level in Denmark. Int J Food Microbiol 2006; 107: Bywater R, Deluyker H, Deroover E et al. A European survey of antimicrobial susceptibility among zoonotic and commensal bacteria isolated from food-producing animals. J Antimicrob Chemother 2004; 54: Desmonts M-H, Dufour-Gesbert F, Avrain L et al. Antimicrobial resistance in Campylobacter strains isolated from French broilers before and after antimicrobial growth promoter bans. J Antimicrob Chemother 2004; 54: Luber P, Wagner J, Hahn H et al. Antimicrobial resistance in Campylobacter jejuni and Campylobacter coli strains isolated in 1991 and from poultry and humans in Berlin, Germany. Antimicrob Agents Chemother 2003; 47: Guevremont E, Dadeau E, Sirois M et al. Antimicrobial susceptibilities of thermophilic Campylobacter from humans, swine and chicken broilers. Can J Vet Res 2006; 70: Luangtongkum T, Morishita TY, Ison AJ et al. Effect of conventional and organic production practices on the prevalence and antimicrobial resistance of Campylobacter spp. in poultry. Appl Environ Microbiol 2006; 72: Manavathu EK, Hiratsuka K, Taylor DE. Nucleotide sequence analysis and expression of a tetracycline-resistance gene from Campylobacter jejuni. Gene 1988; 62: Avrain L, Vernozy-Rozand C, Kempf I. Evidence for natural horizontal transfer of teto gene between Campylobacter jejuni strains in chickens. J Appl Microbiol 2004; 97: Fairchild AS, Smith JL, Idris U et al. Effects of orally administered tetracycline on the intestinal community structure of chickens and on tet determinant carriage by commensal bacteria and Campylobacter jejuni. Appl Environ Microbiol 2005; 71: Lee Y-C, Tai L-C, Lin C-S et al. Occurrence of plasmids and tetracycline resistance among Campylobacter jejuni and Campylobacter coli isolated from whole market chickens and clinical samples. Int J Food Microbiol 1994; 24: Pratt A, Korolik V. Tetracycline resistance of Australian Campylobacter jejuni and Campylobacter coli isolates. J Antimicrob Chemother 2005; 55: Pumbwe L, Piddock LJV. Identification and molecular characterisation of CmeB, a Campylobacter jejuni multidrug efflux pump. FEMS Microbiol Lett 2002; 206: Pumbwe L, Randall LP, Woodward MJ et al. Expression of the efflux pump genes cmeb, cmef and the porin gene pora in multiple-antibiotic-resistant Campylobacter jejuni. J Antimicrob Chemother 2004; 54: Humphrey TJ, Jorgensen F, Frost JA et al. Prevalence and subtypes of ciprofloxacin-resistant Campylobacter spp. in commercial poultry flocks before, during, and after treatment with fluoroquinolones. Antimicrob Agents Chemother 2005; 49: Griggs DJ, Johnson MM, Frost JA et al. Incidence and mechanism of ciprofloxacin resistance in Campylobacter spp. isolated from commercial poultry flocks in the United Kingdom before, during, and after fluoroquinolone treatment. Antimicrob Agents Chemother 2005; 49: Bolton FJ, Wareing DRA, Skirrow MB et al. Identification and biotyping of campylobacters. In: Board GR, Jones D, Skinner FA, eds. Identification Methods in Applied and Environmental Microbiology. Oxford: Blackwell Scientific Publications, 1992; Best EL, Powell EJ, Swift C et al. Applicability of a rapid duplex real-time PCR assay for speciation of Campylobacter jejuni and Campylobacter coli directly from culture plates. FEMS Microbiol Lett 2003; 229: Houf K, Tutenel A, De Zutter L et al. Development of a multiplex PCR assay for the simultaneous detection and identification of Arcobacter butzleri, Arcobacter cryaerophilus and Arcobacter skirrowii. FEMS Microbiol Lett 2000; 193: Nachamkin I, Bohachick K, Patton CM. Flagellin gene typing of Campylobacter jejuni by restriction fragment length polymorphism analysis. J Clin Microbiol 1993; 31: Meinersmann RJ, Helsel LO, Fields PI et al. Discrimination of Campylobacter jejuni isolates by fla gene sequencing. J Clin Microbiol 1997; 35: Gibson JR, Sutherland K, Owen RJ. Inhibition of DNAse activity in PFGE analysis of DNA from Campylobacter jejuni. Lett Appl Microbiol 1994; 19: Thwaites RT, Frost JA. Drug resistance in Campylobacter jejuni, Campylobacter coli, and Campylobacter lari from humans in north west England and Wales, J Clin Pathol 1999; 52:

13 Antibiotic resistance in chlortetracycline-treated poultry flocks 26. McDermott PF, Bodeis SM, Aarestrup FM et al. Development of a standardized susceptibility test for Campylobacter with quality control ranges for ciprofloxacin, doxifloxacin, erythromycin, gentamicin and meropenem. Microb Drug Res 2004; 10: Gibreel A, Taylor DE. Macrolide resistance in Campylobacter jejuni and Campylobacter coli. J Antimicrob Chemother 2006; 58: Cagliero C, Mouline C, Payot S et al. Involvement of the CmeABC efflux pump in the macrolide resistance of Campylobacter coli. J Antimicrob Chemother 2005; 6: Baucheron S, Tyler S, Boyd D et al. AcrAB-TolC directs efflux-mediated multidrug resistance in Salmonella enterica serovar typhimurium DT104. Antimicrob Agents Chemother 2004; 48:

The National Antimicrobial Resistance Monitoring System (NARMS)

The National Antimicrobial Resistance Monitoring System (NARMS) The National Antimicrobial Resistance Monitoring System (NARMS) Strategic Plan 2012-2016 Table of Contents Background... 2 Mission... 3 Overview of Accomplishments, 1996-2011... 4 Strategic Goals and Objectives...

More information

A baseline survey of antimicrobial resistance in bacteria from selected New Zealand foods, 2009 2010

A baseline survey of antimicrobial resistance in bacteria from selected New Zealand foods, 2009 2010 A baseline survey of antimicrobial resistance in bacteria from New Zealand foods, 2009 2010 MAF Technical Paper No: 2011/53 Helen Heffernan 1, Teck Lok Wong 2, Jennifer Lindsay 2, Barbara Bowen1 and Rosemary

More information

Laboratory Protocols Level 2 Training Course Isolation of thermotolerant Campylobacter from faeces

Laboratory Protocols Level 2 Training Course Isolation of thermotolerant Campylobacter from faeces Global Salm-Surv A global Salmonella surveillance and laboratory support project of the World Health Organization Laboratory Protocols Level 2 Training Course Isolation of thermotolerant Campylobacter

More information

EU Reference Laboratory for E. coli Department of Veterinary Public Health and Food Safety Unit of Foodborne Zoonoses Istituto Superiore di Sanità

EU Reference Laboratory for E. coli Department of Veterinary Public Health and Food Safety Unit of Foodborne Zoonoses Istituto Superiore di Sanità EU Reference Laboratory for E. coli Department of Veterinary Public Health and Food Safety Unit of Foodborne Zoonoses Istituto Superiore di Sanità Inventory of the expertise on molecular typing of Verocytotoxin-producing

More information

PATHOGEN DETECTION SYSTEMS BY REAL TIME PCR. Results Interpretation Guide

PATHOGEN DETECTION SYSTEMS BY REAL TIME PCR. Results Interpretation Guide PATHOGEN DETECTION SYSTEMS BY REAL TIME PCR Results Interpretation Guide Pathogen Detection Systems by Real Time PCR Microbial offers real time PCR based systems for the detection of pathogenic bacteria

More information

Multi-locus sequence typing (MLST) of C. jejuni infections in the United States Patrick Kwan, PhD

Multi-locus sequence typing (MLST) of C. jejuni infections in the United States Patrick Kwan, PhD Multi-locus sequence typing (MLST) of C. jejuni infections in the United States Patrick Kwan, PhD National Campylobacter and Helicobacter Reference Laboratory Enteric Diseases Laboratory Branch Centers

More information

National Antimicrobial Resistance Monitoring System - Enteric Bacteria. A program to monitor antimicrobial resistance in humans and animals

National Antimicrobial Resistance Monitoring System - Enteric Bacteria. A program to monitor antimicrobial resistance in humans and animals National Antimicrobial Resistance Monitoring System - Enteric Bacteria A program to monitor antimicrobial resistance in humans and animals Antimicrobial resistance in foodborne pathogens is an important

More information

Copa-Cogeca s views on Critically Important Antibiotics Miguel Angel Higuera (ASAJA, ES)

Copa-Cogeca s views on Critically Important Antibiotics Miguel Angel Higuera (ASAJA, ES) AHW(14)1987:1 Copa-Cogeca s views on Critically Important Antibiotics Miguel Angel Higuera (ASAJA, ES) Vice-Chair of the Copa-Cogeca Working Party on Animal Health and Welfare London 28 th February 2014

More information

Compensation for Salmonella Enteritidis & Typhimurium to owners in England

Compensation for Salmonella Enteritidis & Typhimurium to owners in England Department for Environment, Food and Rural Affairs Compensation for Salmonella Enteritidis & Typhimurium to owners in England January 2015 Contents 1. Introduction... 1 2. Aims and objectives... 1 3. Legal

More information

(Question Nº EFSA-Q-2006-046) Adopted by The Task Force on 20 February 2007

(Question Nº EFSA-Q-2006-046) Adopted by The Task Force on 20 February 2007 Report of the Task Force on Zoonoses Data Collection including a proposal for a harmonized monitoring scheme of antimicrobial resistance in Salmonella in fowl (Gallus gallus), turkeys, and pigs and Campylobacter

More information

Diagnosis, Treatment and Prevention of Typhoid Fever in the Children of Bangladesh: A Microbiologist s View.

Diagnosis, Treatment and Prevention of Typhoid Fever in the Children of Bangladesh: A Microbiologist s View. Diagnosis, Treatment and Prevention of Typhoid Fever in the Children of Bangladesh: A Microbiologist s View. Samir K Saha, Ph.D. Department of Microbiology Dhaka Shishu Hospital Bangladesh Typhoid Fever:Salmonella

More information

Identification of the VTEC serogroups mainly associated with human infections by conventional PCR amplification of O-associated genes

Identification of the VTEC serogroups mainly associated with human infections by conventional PCR amplification of O-associated genes Identification of the VTEC serogroups mainly associated with human infections by conventional PCR amplification of O-associated genes 1. Aim and field of application The present method concerns the identification

More information

Udbredelse og betydning af resistente bakterier: Fokus på spredning fra dyr til mennesker

Udbredelse og betydning af resistente bakterier: Fokus på spredning fra dyr til mennesker Udbredelse og betydning af resistente bakterier: Fokus på spredning fra dyr til mennesker Frank M. Aarestrup (fmaa@food.dtu.dk) Importance of the food animal reservoir for human health Different estimates

More information

Effects of Antibiotics on Bacterial Growth and Protein Synthesis: Student Laboratory Manual

Effects of Antibiotics on Bacterial Growth and Protein Synthesis: Student Laboratory Manual Effects of Antibiotics on Bacterial Growth and Protein Synthesis: Student Laboratory Manual I. Purpose...1 II. Introduction...1 III. Inhibition of Bacterial Growth Protocol...2 IV. Inhibition of in vitro

More information

Disc Diffusion Susceptibility Methods

Disc Diffusion Susceptibility Methods Disc Diffusion Susceptibility Methods Introduction When a filter paper disc impregnated with a chemical is placed on agar the chemical will diffuse from the disc into the agar. This diffusion will place

More information

BIOSECURITY PROCEDURES IN POULTRY PRODUCTION

BIOSECURITY PROCEDURES IN POULTRY PRODUCTION 1 Annex VII CHAPTER 6.4. BIOSECURITY PROCEDURES IN POULTRY PRODUCTION Article 6.4.1. Introduction This chapter provides recommended biosecurity procedures in poultry production and is not specifically

More information

Green Fluorescent Protein (GFP): Genetic Transformation, Synthesis and Purification of the Recombinant Protein

Green Fluorescent Protein (GFP): Genetic Transformation, Synthesis and Purification of the Recombinant Protein Green Fluorescent Protein (GFP): Genetic Transformation, Synthesis and Purification of the Recombinant Protein INTRODUCTION Green Fluorescent Protein (GFP) is a novel protein produced by the bioluminescent

More information

European Medicines Agency

European Medicines Agency European Medicines Agency July 1996 CPMP/ICH/139/95 ICH Topic Q 5 B Quality of Biotechnological Products: Analysis of the Expression Construct in Cell Lines Used for Production of r-dna Derived Protein

More information

Welcome to Implementing Inquirybased Microbial Project. Veronica Ardi, PhD

Welcome to Implementing Inquirybased Microbial Project. Veronica Ardi, PhD Welcome to Implementing Inquirybased Microbial Project Veronica Ardi, PhD Microbiology Laboratory Courses CourseSmart: ebook resources http://instructors.coursesmart.com/ Microbiology Laboratory Courses

More information

PRIORITY RESEARCH TOPICS

PRIORITY RESEARCH TOPICS PRIORITY RESEARCH TOPICS Understanding all the issues associated with antimicrobial resistance is probably impossible, but it is clear that there are a number of key issues about which we need more information.

More information

Biological Sciences Initiative

Biological Sciences Initiative Biological Sciences Initiative HHMI Student Activities Measuring Antibiotic Resistance Introduction: You might be aware that antibiotics were once thought of as a magic bullet; a nearly perfect drug for

More information

Gene Mapping Techniques

Gene Mapping Techniques Gene Mapping Techniques OBJECTIVES By the end of this session the student should be able to: Define genetic linkage and recombinant frequency State how genetic distance may be estimated State how restriction

More information

ANNUAL REPORT ON STAPHYLOCOCCUS AUREUS BACTERAEMIA CASES IN DENMARK 2008 (part I)

ANNUAL REPORT ON STAPHYLOCOCCUS AUREUS BACTERAEMIA CASES IN DENMARK 2008 (part I) ANNUAL REPORT ON STAPHYLOCOCCUS AUREUS BACTERAEMIA CASES IN DENMARK 2008 (part I) STAPHYLOCOCCUS LABORATORY, STATENS SERUM INSTITUT 1 Staphylococcus aureus bacteraemia annual report, part I The format

More information

Corvinus University of Budapest Faculty of Food Science Department of Microbiology and Biotechnology

Corvinus University of Budapest Faculty of Food Science Department of Microbiology and Biotechnology Corvinus University of Budapest Faculty of Food Science Department of Microbiology and Biotechnology DETECTION, PCR-BASED MOLECULAR IDENTIFICATION AND TYPING OF FOOD SAFETY RELATED BACTERIA Theses Ágnes

More information

Raw Milk Quality Tests Do They Predict Fluid Milk Shelf-life or Is it time for new tests?

Raw Milk Quality Tests Do They Predict Fluid Milk Shelf-life or Is it time for new tests? Raw Milk Quality Tests Do They Predict Fluid Milk Shelf-life or Is it time for new tests? Martin Wiedmann Milk Quality Improvement Program November 3, 2011 Fluid milk shelf life What defines shelf life

More information

Molecular fingerprinting of Campylobacter and Arcobacter isolated from chicken and water

Molecular fingerprinting of Campylobacter and Arcobacter isolated from chicken and water RESEARCH ARTICLE INTERNATIONAL MICROBIOLOGY (2007) 10:85-90 DOI: 10.2436/20.1501.01.12 ISSN: 1139-6709 www.im.microbios.org Molecular fingerprinting of Campylobacter and Arcobacter isolated from chicken

More information

GROWING BACTERIA INTRODUCTION

GROWING BACTERIA INTRODUCTION GROWING BACTERIA INTRODUCTION E. coli is a normal part of the bacterial flora of the human gut. It is not generally considered pathogenic, although some strains are highly toxic (recent food poisonings

More information

Intended Use: The kit is designed to detect the 5 different mutations found in Asian population using seven different primers.

Intended Use: The kit is designed to detect the 5 different mutations found in Asian population using seven different primers. Unzipping Genes MBPCR014 Beta-Thalassemia Detection Kit P r o d u c t I n f o r m a t i o n Description: Thalassemia is a group of genetic disorders characterized by quantitative defects in globin chain

More information

www.biochemj.org/bj/330/0581/bj3300581.htm

www.biochemj.org/bj/330/0581/bj3300581.htm Ribosomes as Antibiotic Targets www.biochemj.org/bj/330/0581/bj3300581.htm Ware, Bioscience in the 21 st Century, 2009 PERSPECTIVE Widespread use of antibiotics after WWII improved human health globally

More information

Antimicrobial Resistance and Human Health

Antimicrobial Resistance and Human Health Antimicrobial Resistance and Human Health Dearbháile Morris, Discipline of Bacteriology, School of Medicine, National University of Ireland, Galway The microbial world The is a gene Talk cloud in a The

More information

NS5B Sequencing and Phenotypic Resistance Assays for HCV Subtypes 1a and 1b

NS5B Sequencing and Phenotypic Resistance Assays for HCV Subtypes 1a and 1b NS5B Sequencing and Phenotypic Resistance Assays for HCV Subtypes 1a and 1b 5th Intl. Workshop on Hepatitis C Resistance & New Compounds Jacqueline Reeves NS5B Resistance Assays for HCV Subtypes 1a and

More information

C ampylobacters were first isolated from humans in 1938

C ampylobacters were first isolated from humans in 1938 749 ORIGINAL ARTICLE Detection of campylobacter species: a comparison of culture and polymerase chain reaction based methods S P Kulkarni, S Lever, JMJLogan, A J Lawson, J Stanley, M S Shafi... J Clin

More information

ANIMAL ANTIBIOTICS: Keeping Animals Healthy and Our Food Safe

ANIMAL ANTIBIOTICS: Keeping Animals Healthy and Our Food Safe ANIMAL ANTIBIOTICS: Keeping Animals Healthy and Our Food Safe Protecting Animal Health To keep animals healthy, veterinarians and farmers work together to create flock and herd healthmanagement programs

More information

Statement by Jay P. Graham, PhD, MBA Research Fellow at the Johns Hopkins Bloomberg School of Public Health

Statement by Jay P. Graham, PhD, MBA Research Fellow at the Johns Hopkins Bloomberg School of Public Health Statement by Jay P. Graham, PhD, MBA Research Fellow at the Johns Hopkins Bloomberg School of Public Health Good morning Mr. Chairman and Members of the Senate Health, Education, Labor and Pensions Committee.

More information

CCR Biology - Chapter 9 Practice Test - Summer 2012

CCR Biology - Chapter 9 Practice Test - Summer 2012 Name: Class: Date: CCR Biology - Chapter 9 Practice Test - Summer 2012 Multiple Choice Identify the choice that best completes the statement or answers the question. 1. Genetic engineering is possible

More information

LECTURE 6 Gene Mutation (Chapter 16.1-16.2)

LECTURE 6 Gene Mutation (Chapter 16.1-16.2) LECTURE 6 Gene Mutation (Chapter 16.1-16.2) 1 Mutation: A permanent change in the genetic material that can be passed from parent to offspring. Mutant (genotype): An organism whose DNA differs from the

More information

ANNUAL REPORT ON STAPHYLOCOCCUS AUREUS BACTERAEMIA CASES IN DENMARK 2009 (part I)

ANNUAL REPORT ON STAPHYLOCOCCUS AUREUS BACTERAEMIA CASES IN DENMARK 2009 (part I) ANNUAL REPORT ON STAPHYLOCOCCUS AUREUS BACTERAEMIA CASES IN DENMARK 2009 (part I) STAPHYLOCOCCUS LABORATORY, STATENS SERUM INSTITUT 1 Staphylococcus aureus Bacteraemia Annual Report, Part I The annual

More information

Facts about the production of Poultry Meat in Denmark 4. July 2014

Facts about the production of Poultry Meat in Denmark 4. July 2014 Facts about the production of Poultry Meat in Denmark 4. July 2014 Birthe Steenberg Manager Danish Poultry Meat Association Tlf. 24631673; E-mail: bsb@lf.dk Poultry Meat from stable to table Breeding animals

More information

QUANTIFICATION OF CAMPYLOBACTER FROM INTERNAL AND EXTERNAL CARCASS RINSES. FINAL REPORT

QUANTIFICATION OF CAMPYLOBACTER FROM INTERNAL AND EXTERNAL CARCASS RINSES. FINAL REPORT QUANTIFICATION OF CAMPYLOBACTER FROM INTERNAL AND EXTERNAL CARCASS RINSES. FINAL REPORT Prepared as part of a New Zealand Food Safety Authority contract for scientific services by Dr Susan Paulin Dr Tecklok

More information

INTERNATIONAL CONFERENCE ON HARMONISATION OF TECHNICAL REQUIREMENTS FOR REGISTRATION OF PHARMACEUTICALS FOR HUMAN USE Q5B

INTERNATIONAL CONFERENCE ON HARMONISATION OF TECHNICAL REQUIREMENTS FOR REGISTRATION OF PHARMACEUTICALS FOR HUMAN USE Q5B INTERNATIONAL CONFERENCE ON HARMONISATION OF TECHNICAL REQUIREMENTS FOR REGISTRATION OF PHARMACEUTICALS FOR HUMAN USE ICH HARMONISED TRIPARTITE GUIDELINE QUALITY OF BIOTECHNOLOGICAL PRODUCTS: ANALYSIS

More information

Lab 10: Bacterial Transformation, part 2, DNA plasmid preps, Determining DNA Concentration and Purity

Lab 10: Bacterial Transformation, part 2, DNA plasmid preps, Determining DNA Concentration and Purity Lab 10: Bacterial Transformation, part 2, DNA plasmid preps, Determining DNA Concentration and Purity Today you analyze the results of your bacterial transformation from last week and determine the efficiency

More information

2. Incidence, prevalence and duration of breastfeeding

2. Incidence, prevalence and duration of breastfeeding 2. Incidence, prevalence and duration of breastfeeding Key Findings Mothers in the UK are breastfeeding their babies for longer with one in three mothers still breastfeeding at six months in 2010 compared

More information

restriction enzymes 350 Home R. Ward: Spring 2001

restriction enzymes 350 Home R. Ward: Spring 2001 restriction enzymes 350 Home Restriction Enzymes (endonucleases): molecular scissors that cut DNA Properties of widely used Type II restriction enzymes: recognize a single sequence of bases in dsdna, usually

More information

EU Reference Laboratory for E. coli Department of Veterinary Public Health and Food Safety Unit of Foodborne Zoonoses Istituto Superiore di Sanità

EU Reference Laboratory for E. coli Department of Veterinary Public Health and Food Safety Unit of Foodborne Zoonoses Istituto Superiore di Sanità Identification and characterization of Verocytotoxin-producing Escherichia coli (VTEC) by Real Time PCR amplification of the main virulence genes and the genes associated with the serogroups mainly associated

More information

NORWAY TRENDS AND SOURCES OF ZOONOSES AND ZOONOTIC AGENTS IN HUMANS, FOODSTUFFS ANIMALS AND FEEDINGSTUFF

NORWAY TRENDS AND SOURCES OF ZOONOSES AND ZOONOTIC AGENTS IN HUMANS, FOODSTUFFS ANIMALS AND FEEDINGSTUFF NORWAY TRENDS AND SOURCES OF ZOONOSES AND ZOONOTIC AGENTS IN HUMANS, FOODSTUFFS ANIMALS AND FEEDINGSTUFF - including information on foodborne outbreaks, antimicrobial resistance in zoonotic agents and

More information

Forensic DNA Testing Terminology

Forensic DNA Testing Terminology Forensic DNA Testing Terminology ABI 310 Genetic Analyzer a capillary electrophoresis instrument used by forensic DNA laboratories to separate short tandem repeat (STR) loci on the basis of their size.

More information

The 11 th EURL-AR Proficiency Testing Salmonella and Campylobacter 2011. Susanne Karlsmose Lina M. Cavaco Rene S. Hendriksen Frank M.

The 11 th EURL-AR Proficiency Testing Salmonella and Campylobacter 2011. Susanne Karlsmose Lina M. Cavaco Rene S. Hendriksen Frank M. The 11 th EURL-AR Proficiency Testing Salmonella and Campylobacter 2011 Susanne Karlsmose Lina M. Cavaco Rene S. Hendriksen Frank M. Aarestrup European Union Reference Laboratory - Antimicrobial Resistance

More information

Integrated Surveillance of Antimicrobial Resistance

Integrated Surveillance of Antimicrobial Resistance ii iii WORLD HEALTH ORGANIZATION Integrated Surveillance of Antimicrobial Resistance Guidance from a WHO Advisory Group World Health Organization, Geneva iv WHO Library Cataloguing-in-Publication Data

More information

The 14th EURL-AR Proficiency Test - enterococci, staphylococci and E. coli 2013. Lina Cavaco Susanne Karlsmose Rene S. Hendriksen Frank M.

The 14th EURL-AR Proficiency Test - enterococci, staphylococci and E. coli 2013. Lina Cavaco Susanne Karlsmose Rene S. Hendriksen Frank M. The 14th EURL-AR Proficiency Test - enterococci, staphylococci and E. coli 2013 Lina Cavaco Susanne Karlsmose Rene S. Hendriksen Frank M. Aarestrup The 14TH EURL-AR Proficiency Test Enterococci, Staphylococci

More information

AMES TEST: Bacterial Reverse Mutation Assay

AMES TEST: Bacterial Reverse Mutation Assay AMES TEST: Bacterial Reverse Mutation Assay 1. Introduction The bacteria reversed mutation assay (Ames Test) is used to evaluate the mutagenic properties of test articles. The test uses amino acid-dependent

More information

Lecture 13: DNA Technology. DNA Sequencing. DNA Sequencing Genetic Markers - RFLPs polymerase chain reaction (PCR) products of biotechnology

Lecture 13: DNA Technology. DNA Sequencing. DNA Sequencing Genetic Markers - RFLPs polymerase chain reaction (PCR) products of biotechnology Lecture 13: DNA Technology DNA Sequencing Genetic Markers - RFLPs polymerase chain reaction (PCR) products of biotechnology DNA Sequencing determine order of nucleotides in a strand of DNA > bases = A,

More information

BCOR101 Midterm II Wednesday, October 26, 2005

BCOR101 Midterm II Wednesday, October 26, 2005 BCOR101 Midterm II Wednesday, October 26, 2005 Name Key Please show all of your work. 1. A donor strain is trp+, pro+, met+ and a recipient strain is trp-, pro-, met-. The donor strain is infected with

More information

TransformAid Bacterial Transformation Kit

TransformAid Bacterial Transformation Kit Home Contacts Order Catalog Support Search Alphabetical Index Numerical Index Restriction Endonucleases Modifying Enzymes PCR Kits Markers Nucleic Acids Nucleotides & Oligonucleotides Media Transfection

More information

CMT - Copan Milk Test

CMT - Copan Milk Test - Copan Milk Test for the detection of Antibiotics & Sulphonamides in milk and milk products - Copan Milk Test Copan Milk Test () Description and principle of the test is available as a single test which

More information

CHAPTER 6: RECOMBINANT DNA TECHNOLOGY YEAR III PHARM.D DR. V. CHITRA

CHAPTER 6: RECOMBINANT DNA TECHNOLOGY YEAR III PHARM.D DR. V. CHITRA CHAPTER 6: RECOMBINANT DNA TECHNOLOGY YEAR III PHARM.D DR. V. CHITRA INTRODUCTION DNA : DNA is deoxyribose nucleic acid. It is made up of a base consisting of sugar, phosphate and one nitrogen base.the

More information

The Techniques of Molecular Biology: Forensic DNA Fingerprinting

The Techniques of Molecular Biology: Forensic DNA Fingerprinting Revised Fall 2011 The Techniques of Molecular Biology: Forensic DNA Fingerprinting The techniques of molecular biology are used to manipulate the structure and function of molecules such as DNA and proteins

More information

3.0 Treatment of Infection

3.0 Treatment of Infection 3.0 Treatment of Infection Antibiotics and Medicine National Curriculum Link SCN 3-13b SCN 3-20b HWB 3-15a HWB 3-16a HWB 3-17a Learning Outcomes All students will know: Most common infections will get

More information

Sampling and the Health Protection Agency

Sampling and the Health Protection Agency Food Water and Environmental Microbiology Sampling and the Health Protection Agency Rob Johnston Health Protection Agency http://www.hpa.org.uk Health Protection Agency - Services Health Protection Services

More information

Introduction To Real Time Quantitative PCR (qpcr)

Introduction To Real Time Quantitative PCR (qpcr) Introduction To Real Time Quantitative PCR (qpcr) SABiosciences, A QIAGEN Company www.sabiosciences.com The Seminar Topics The advantages of qpcr versus conventional PCR Work flow & applications Factors

More information

Cystic Fibrosis Webquest Sarah Follenweider, The English High School 2009 Summer Research Internship Program

Cystic Fibrosis Webquest Sarah Follenweider, The English High School 2009 Summer Research Internship Program Cystic Fibrosis Webquest Sarah Follenweider, The English High School 2009 Summer Research Internship Program Introduction: Cystic fibrosis (CF) is an inherited chronic disease that affects the lungs and

More information

GENE CLONING AND RECOMBINANT DNA TECHNOLOGY

GENE CLONING AND RECOMBINANT DNA TECHNOLOGY GENE CLONING AND RECOMBINANT DNA TECHNOLOGY What is recombinant DNA? DNA from 2 different sources (often from 2 different species) are combined together in vitro. Recombinant DNA forms the basis of cloning.

More information

Enteric Septicemia of Catfish

Enteric Septicemia of Catfish Enteric Septicemia of Catfish Jesse Chappell Extension Fisheries Specialist Revised 2008 Enteric Septicemia of Catfish (ESC) has become one of the two most significant diseases of economic significance

More information

CALL FOR DATA ON VEROTOXIGENIC ESCHERICHIA COLI (VTEC) / SHIGATOXIGENIC E. COLI (STEC)

CALL FOR DATA ON VEROTOXIGENIC ESCHERICHIA COLI (VTEC) / SHIGATOXIGENIC E. COLI (STEC) Joint FAO/WHO Expert Meetings on Microbiological Risk Assessment (JEMRA) CALL FOR DATA ON VEROTOXIGENIC ESCHERICHIA COLI (VTEC) / SHIGATOXIGENIC E. COLI (STEC) Background Deadline: 17 June 2016 Verotoxigenic

More information

Public health and safety of eggs and egg products in Australia. Explanatory summary of the risk assessment

Public health and safety of eggs and egg products in Australia. Explanatory summary of the risk assessment Public health and safety of eggs and egg products in Australia Explanatory summary of the risk assessment FOOD STANDARDS Australia New Zealand Public health and safety of eggs and egg products in Australia

More information

Microbiology BIOL 275 DILUTIONS

Microbiology BIOL 275 DILUTIONS DILUTIONS Occasionally a solution is too concentrated to be used as is. For example, when one is performing manual blood counts, the blood contains too many cells to be counted as such. Or when performing

More information

Healthy California 2020 Initiative: Consensus Building on Top Priority Areas for CDPH

Healthy California 2020 Initiative: Consensus Building on Top Priority Areas for CDPH Healthy California 2020 Initiative: Consensus Building on Top Priority Areas for CDPH Public Health Advisory Committee April 30, 2010 Introducing the CDPH Decision Framework Responding to public health

More information

ID kit. imegen Anchovies II. and E. japonicus) DNA detection by. User manual. Anchovies species (E. encrasicolus. sequencing.

ID kit. imegen Anchovies II. and E. japonicus) DNA detection by. User manual. Anchovies species (E. encrasicolus. sequencing. User manual imegen Anchovies II ID kit Anchovies species (E. encrasicolus and E. japonicus) DNA detection by sequencing Reference: Made in Spain The information in this guide is subject to change without

More information

Data Analysis for Ion Torrent Sequencing

Data Analysis for Ion Torrent Sequencing IFU022 v140202 Research Use Only Instructions For Use Part III Data Analysis for Ion Torrent Sequencing MANUFACTURER: Multiplicom N.V. Galileilaan 18 2845 Niel Belgium Revision date: August 21, 2014 Page

More information

Research Project Summary

Research Project Summary Division of Science, Research and Technology Research Project Summary June, 25 Microbial Source Tracking in the Manasquan River Estuary Michael A. Palladino, Ph.D. and John A. Tiedemann Thomas B. Atherholt,

More information

Introduction. Introduction. Why do we need microbiological diagnostics of udder infections? Microbiological diagnostics How is it done?

Introduction. Introduction. Why do we need microbiological diagnostics of udder infections? Microbiological diagnostics How is it done? Introduction Microbiological diagnostics of udder infections Karin Persson Waller National Veterinary Institute (SVA) Swedish University of Agricultural Sciences Uppsala, Sweden Mastitis = in most cases

More information

First Strand cdna Synthesis

First Strand cdna Synthesis 380PR 01 G-Biosciences 1-800-628-7730 1-314-991-6034 technical@gbiosciences.com A Geno Technology, Inc. (USA) brand name First Strand cdna Synthesis (Cat. # 786 812) think proteins! think G-Biosciences

More information

Food Safety Performance

Food Safety Performance Food Safety Performance Introduction The GeoRisQ Food Safety Performance Monitor has been developed within the context of the report Food Safety in Europe: From Farm to Fork and Further. This report is

More information

Nucleic Acid Purity Assessment using A 260 /A 280 Ratios

Nucleic Acid Purity Assessment using A 260 /A 280 Ratios Nucleic Acid Purity Assessment using A 260 /A 280 Ratios A common practice in molecular biology is to perform a quick assessment of the purity of nucleic acid samples by determining the ratio of spectrophotometric

More information

MTT Cell Proliferation Assay

MTT Cell Proliferation Assay ATCC 30-1010K Store at 4 C This product is intended for laboratory research purposes only. It is not intended for use in humans, animals or for diagnostics. Introduction Measurement of cell viability and

More information

2013 Indiana Healthcare Provider and Hospital Administrator Multi-Drug Resistant Organism Survey

2013 Indiana Healthcare Provider and Hospital Administrator Multi-Drug Resistant Organism Survey 2013 Indiana Healthcare Provider and Hospital Administrator Multi-Drug Resistant Organism Survey Antibiotic resistance is a global issue that has significant impact in the field of infectious diseases.

More information

Management of Extended Spectrum Beta- Lactamase (ESBL) Producing Enterobacteriaceae in health care settings

Management of Extended Spectrum Beta- Lactamase (ESBL) Producing Enterobacteriaceae in health care settings Management of Extended Spectrum Beta- Lactamase (ESBL) Producing Enterobacteriaceae in health care settings Dr. Mary Vearncombe PIDAC-IPC February 2012 Objectives: To provide an overview of the RP/AP Annex

More information

MDM. Metabolic Drift Mutations - Attenuation Technology

MDM. Metabolic Drift Mutations - Attenuation Technology MDM Metabolic Drift Mutations - Attenuation Technology Seite 2 Origin of MDM attenuation technology Prof. Dr. Klaus Linde Pioneer in R&D of human and animal vaccines University of Leipzig Germany Origin

More information

Molecular Biology Techniques: A Classroom Laboratory Manual THIRD EDITION

Molecular Biology Techniques: A Classroom Laboratory Manual THIRD EDITION Molecular Biology Techniques: A Classroom Laboratory Manual THIRD EDITION Susan Carson Heather B. Miller D.Scott Witherow ELSEVIER AMSTERDAM BOSTON HEIDELBERG LONDON NEW YORK OXFORD PARIS SAN DIEGO SAN

More information

Original Article Clinical profile and antibiotics response in typhoid fever

Original Article Clinical profile and antibiotics response in typhoid fever Kathmandu University Medical Journal (6), Vol. 4, No. 1, Issue 13, 25-29 Original Article Clinical profile and antibiotics response in typhoid fever Bajracharya BL 1, Baral MR 2, Shakya S 3, Tuladhar P

More information

Transmission of genetic variation: conjugation. Transmission of genetic variation: conjugation

Transmission of genetic variation: conjugation. Transmission of genetic variation: conjugation Transmission of genetic variation: conjugation Transmission of genetic variation: conjugation Bacterial Conjugation is genetic recombination in which there is a transfer of DNA from a living donor bacterium

More information

Whole genome sequencing of foodborne pathogens: experiences from the Reference Laboratory. Kathie Grant Gastrointestinal Bacteria Reference Unit

Whole genome sequencing of foodborne pathogens: experiences from the Reference Laboratory. Kathie Grant Gastrointestinal Bacteria Reference Unit Whole genome sequencing of foodborne pathogens: experiences from the Reference Laboratory Kathie Grant Gastrointestinal Bacteria Reference Unit 16 th June 2014 Planning for Implementation of WGS 2011-2014

More information

Co Extra (GM and non GM supply chains: Their CO EXistence and TRAceability) Outcomes of Co Extra

Co Extra (GM and non GM supply chains: Their CO EXistence and TRAceability) Outcomes of Co Extra GM and non GM supply chains: Their CO EXistence and TRAceability Outcomes of Co Extra Comparison of different real time PCR chemistries and their suitability for detection and quantification of genetically

More information

Sur les traces du Campylobacter au Luxembourg

Sur les traces du Campylobacter au Luxembourg Environmental sources of Campylobacter infections in Luxembourg Priorité Nationale C09/BM/09 Sur les traces du Campylobacter au Luxembourg Catherine Ragimbeau What sort of germ is Campylobacter? A fragile

More information

TURIN. Historical capital of Italy. City of Art, Nature, Food and Sport. Turin is crossed by the Po river, the Italy s longest river

TURIN. Historical capital of Italy. City of Art, Nature, Food and Sport. Turin is crossed by the Po river, the Italy s longest river TURIN Historical capital of Italy City of Art, Nature, Food and Sport Turin is crossed by the Po river, the Italy s longest river The Mole Antonelliana (1863-1889), 167.5 Meters tall is the symbol of the

More information

TEST METHOD VERIFICATION AND VALIDATION

TEST METHOD VERIFICATION AND VALIDATION 1 TEST METHOD VERIFICATION AND VALIDATION SWACM 2014 MICHAEL LOEFFELHOLZ, PH.D., ABMM DEPT. PATHOLOGY UNIV TEXAS MEDICAL BRANCH GALVESTON, TX 2 OBJECTIVES Define validation and verification Describe components

More information

skin and soft tissue infections (skinfold pyoderma, impetigo, folliculitis, furunculosis, cellulitis) caused by susceptible strains of organisms.

skin and soft tissue infections (skinfold pyoderma, impetigo, folliculitis, furunculosis, cellulitis) caused by susceptible strains of organisms. Part II SUMMARY OF PRODUCT CHARACTERISTICS 1. NAME OF THE VETERINARY MEDICINAL PRODUCT MARBOCYL P 5 mg tablet 2. QUALITATIVE AND QUANTITATIVE COMPOSITION Active Ingredient Marbofloxacin 5mg per tablet

More information

A and B are not absolutely linked. They could be far enough apart on the chromosome that they assort independently.

A and B are not absolutely linked. They could be far enough apart on the chromosome that they assort independently. Name Section 7.014 Problem Set 5 Please print out this problem set and record your answers on the printed copy. Answers to this problem set are to be turned in to the box outside 68-120 by 5:00pm on Friday

More information

BacReady TM Multiplex PCR System

BacReady TM Multiplex PCR System BacReady TM Multiplex PCR System Technical Manual No. 0191 Version 10112010 I Description.. 1 II Applications 2 III Key Features.. 2 IV Shipping and Storage. 2 V Simplified Procedures. 2 VI Detailed Experimental

More information

MGC premier full length cdna and ORF clones

MGC premier full length cdna and ORF clones MGC premier full length cdna and ORF clones TCH1003, TCM1004, TCR1005, TCB1006, TCL1007, TCT1008, TCZ1009, TOH6003, TOM6004, TOZ6009, TCHS1003, TCMS1004, TCRS1005, TCBS1006, TCLS1007, TCTS1008 MGC premier

More information

CLONING IN ESCHERICHIA COLI

CLONING IN ESCHERICHIA COLI CLONING IN ESCHERICHIA COLI Introduction: In this laboratory, you will carry out a simple cloning experiment in E. coli. Specifically, you will first create a recombinant DNA molecule by carrying out a

More information

DNA Technology Mapping a plasmid digesting How do restriction enzymes work?

DNA Technology Mapping a plasmid digesting How do restriction enzymes work? DNA Technology Mapping a plasmid A first step in working with DNA is mapping the DNA molecule. One way to do this is to use restriction enzymes (restriction endonucleases) that are naturally found in bacteria

More information

Gene Expression Assays

Gene Expression Assays APPLICATION NOTE TaqMan Gene Expression Assays A mpl i fic ationef ficienc yof TaqMan Gene Expression Assays Assays tested extensively for qpcr efficiency Key factors that affect efficiency Efficiency

More information

Statistical estimation using confidence intervals

Statistical estimation using confidence intervals 0894PP_ch06 15/3/02 11:02 am Page 135 6 Statistical estimation using confidence intervals In Chapter 2, the concept of the central nature and variability of data and the methods by which these two phenomena

More information

2. True or False? The sequence of nucleotides in the human genome is 90.9% identical from one person to the next. False (it s 99.

2. True or False? The sequence of nucleotides in the human genome is 90.9% identical from one person to the next. False (it s 99. 1. True or False? A typical chromosome can contain several hundred to several thousand genes, arranged in linear order along the DNA molecule present in the chromosome. True 2. True or False? The sequence

More information

Essentials of Real Time PCR. About Sequence Detection Chemistries

Essentials of Real Time PCR. About Sequence Detection Chemistries Essentials of Real Time PCR About Real-Time PCR Assays Real-time Polymerase Chain Reaction (PCR) is the ability to monitor the progress of the PCR as it occurs (i.e., in real time). Data is therefore collected

More information

Personal Injury TYPES OF HOLIDAY ILLNESSES. www.simpsonmillar.co.uk Telephone 0844 858 3200

Personal Injury TYPES OF HOLIDAY ILLNESSES. www.simpsonmillar.co.uk Telephone 0844 858 3200 TYPES OF HOLIDAY ILLNESSES Whilst on holiday many different contractable illnesses exist, the list below contains the most common. This list is by no means exhaustive and if you have suffered from an illness

More information

ABSTRACT. Promega Corporation, Updated September 2008. http://www.promega.com/pubhub. 1 Campbell-Staton, S.

ABSTRACT. Promega Corporation, Updated September 2008. http://www.promega.com/pubhub. 1 Campbell-Staton, S. A Modified Wizard SV Genomic DNA Purification System Protocol to Purify Genomic DNA... A Modified Wizard SV Genomic DNA Purification System Protocol to Purify Genomic DNA from Shed Reptile Skin ABSTRACT

More information

Real-time quantitative RT -PCR (Taqman)

Real-time quantitative RT -PCR (Taqman) Real-time quantitative RT -PCR (Taqman) Author: SC, Patti Lab, 3/03 This is performed as a 2-step reaction: 1. cdna synthesis from DNase 1-treated total RNA 2. PCR 1. cdna synthesis (Advantage RT-for-PCR

More information

The Antimicrobial Susceptibility Testing Device Manufacturer: Development Process, Timelines, and Challenges

The Antimicrobial Susceptibility Testing Device Manufacturer: Development Process, Timelines, and Challenges The Antimicrobial Susceptibility Testing Device Manufacturer: Development Process, Timelines, and Challenges Sheila Farnham, MT(ASCP) President Susceptibility Testing Manufacturers Association January

More information