Comparison of chicken gut colonisation by the pathogens Campylobacter jejuni and Clostridium perfringens by real-time quantitative PCR

被引:80
作者
Skanseng, Beate
Kaldhusdal, Magne
Rudi, Knut
机构
[1] Norwegian Food Res Inst, MATFORSK, N-1430 As, Norway
[2] Norwegian Univ Life Sci, Dept Chem Biotechnol & Food Sci, N-1432 As, Norway
[3] Natl Vet Inst, N-0033 Oslo, Norway
[4] Hedmark Univ Coll, Fac Educ & Sci, N-2306 Hamar, Norway
关键词
Campylobacter jejuni; Clostridium petfringens; quantification; real-time PCR; chicken; caecum;
D O I
10.1016/j.mcp.2006.02.001
中图分类号
Q5 [生物化学];
学科分类号
071010 [生物化学与分子生物学]; 081704 [应用化学];
摘要
We compared the colonisation of the chicken gut by the two important pathogens Campylobacter jejuni (frequent food-borne pathogen) and alpha-toxin gene containing Clostridium perfringens (causative agent of necrotic enteritis in chickens) using a new high-throughput automated DNA purification method for microbial biodiversity analyses. The method gave high reproducibility (standard deviation of 1.1 C-T-values for a universal 16S rDNA real-time PCR), and inhibition was observed in only 0.9% of the individual DNA purifications (n = 753). We analysed 253 randomly collected chicken caecal samples (sampled in 2001 and 2003) from Norwegian chicken flocks by real-time quantitative PCR. Our results showed positive correlation (P = 0.009) in chicken caccal colonisation between C jejuni and Cl. perfringens. We also found that there was a significant underrepresentation (P = 0.008) of chickens containing high levels of Cl. petfringens and low levels of C jejuni. This indicates a possible interaction between these bacteria. Potential interaction between pathogens and other bacteria in the gut will certainly be important research fields in the future. As demonstrated here, the development of new tools for high-throughput analyses will be of key importance for these studies. (c) 2006 Elsevier Ltd. All rights reserved.
引用
收藏
页码:269 / 279
页数:11
相关论文
共 27 条
[1]
Divergence and redundancy of 16S rRNA sequences in genomes with multiple rrn operons [J].
Acinas, SG ;
Marcelino, LA ;
Klepac-Ceraj, V ;
Polz, MF .
JOURNAL OF BACTERIOLOGY, 2004, 186 (09) :2629-2635
[2]
Apajalahti JHA, 1998, APPL ENVIRON MICROB, V64, P4084
[3]
Contamination and sensitivity issues with a real-time universal 16S rRNA PCR [J].
Corless, CE ;
Guiver, M ;
Borrow, R ;
Edwards-Jones, V ;
Kaczmarski, EB ;
Fox, AJ .
JOURNAL OF CLINICAL MICROBIOLOGY, 2000, 38 (05) :1747-1752
[4]
What has happened in Norway after the ban of avoparcin? Consumption of antimicrobials by poultry [J].
Grave, K ;
Kaldhusdal, M ;
Kruse, H ;
Harr, LMF ;
Flatlandsmo, K .
PREVENTIVE VETERINARY MEDICINE, 2004, 62 (01) :59-72
[5]
Extensive profiling of a complex microbial community by high-throughput sequencing [J].
Hill, JE ;
Seipp, RP ;
Betts, M ;
Hawkins, L ;
Van Kessel, AG ;
Crosby, WL ;
Hemmingsen, SM .
APPLIED AND ENVIRONMENTAL MICROBIOLOGY, 2002, 68 (06) :3055-3066
[6]
IDENTIFICATION AND ELIMINATION OF DNA-SEQUENCES IN TAQ DNA-POLYMERASE [J].
HUGHES, MS ;
BECK, LA ;
SKUCE, RA .
JOURNAL OF CLINICAL MICROBIOLOGY, 1994, 32 (08) :2007-2008
[7]
Reduced incidence of Clostridium perfringens-associated lesions and improved performance in broiler chickens treated with normal intestinal bacteria from adult fowl [J].
Kaldhusdal, M ;
Schneitz, C ;
Hofshagen, M ;
Skjerv, E .
AVIAN DISEASES, 2001, 45 (01) :149-156
[8]
Comparison of automated and manual nucleic acid extraction methods for detection of enterovirus RNA [J].
Knepp, JH ;
Geahr, MA ;
Forman, MS ;
Valsamakis, A .
JOURNAL OF CLINICAL MICROBIOLOGY, 2003, 41 (08) :3532-3536
[9]
UTILIZATION OF HYDROGEN AND FORMATE BY CAMPYLOBACTER SPEC UNDER AEROBIC AND ANAEROBIC CONDITIONS [J].
LAANBROEK, HJ ;
STAL, LJ ;
VELDKAMP, H .
ARCHIVES OF MICROBIOLOGY, 1978, 119 (01) :99-102
[10]
Liver lesions seen at slaughter as an indicator of necrotic enteritis in broiler flocks [J].
Lovland, A ;
Kaldhusdal, M .
FEMS IMMUNOLOGY AND MEDICAL MICROBIOLOGY, 1999, 24 (03) :345-351