Reichelt Benjamin, Szott Vanessa, Stingl Kerstin, Roesler Uwe, Friese Anika
Institute for Animal Hygiene and Environmental Health, Freie Universität Berlin, 14163 Berlin, Germany.
Institute of Food Safety and Food Hygiene, Freie Universität Berlin, 14163 Berlin, Germany.
Microorganisms. 2023 Oct 4;11(10):2492. doi: 10.3390/microorganisms11102492.
Campylobacteriosis cases in humans are of global concern, with high prevalence rates in the poultry reservoir considered the most important source of infection. Research findings show ' ability to enter a viable but non-culturable (VBNC) state, remaining "viable" but unable to grow on culture media. We explored the persistence of VBNC states in specific environments, particularly at broiler farms, as this state may lead to an underestimation of the present prevalence. For VBNC detection, a propidium monoazide PMA-dye viability qPCR (v-qPCR) was used in combination with cultivation methods. We examined samples collected from broiler farm barns and their surroundings, as well as chicken manure from experimental pens. In addition, the tenacity of culturable and VBNC- was studied in vitro in soil and water. In a total of three visits, was not detected either culturally or by v-qPCR (no DNA) in the environment of the broiler farms. In four visits, however, VBNC- were detected both inside and outside the barns. The overall prevalence in environmental samples was 15.9% for VBNC-, 62.2% for DNA, and 1.2% for culturable . In the experimental pens, no cultivable was detected in chicken manure after 24 h. Strikingly, "VBNC-" persisted even after 72 h. "VBNC-" were confirmed in barn surroundings and naturally contaminated chicken manure. Laboratory studies revealed that VBNC- can remain intact in soil for up to 28 days and in water for at least 63 days, depending on environmental conditions.
人类弯曲杆菌病病例受到全球关注，在家禽宿主中高流行率被认为是最重要的感染源。研究结果表明，弯曲杆菌能够进入活的非可培养（VBNC）状态，保持“存活”但无法在培养基上生长。我们探讨了VBNC状态在特定环境中的持续性，特别是在肉鸡养殖场，因为这种状态可能导致对当前流行率的低估。对于VBNC检测，使用单叠氮碘化丙啶（PMA）染料活力定量聚合酶链反应（v-qPCR）结合培养方法。我们检查了从肉鸡养殖舍及其周围环境采集的样本，以及来自实验围栏的鸡粪。此外，还在体外研究了可培养菌和VBNC菌在土壤和水中的存活能力。总共进行了三次走访，在肉鸡养殖场环境中，无论是通过培养还是v-qPCR都未检测到弯曲杆菌（无弯曲杆菌DNA）。然而，在四次走访中，在养殖舍内外都检测到了VBNC弯曲杆菌。环境样本中VBNC弯曲杆菌的总体流行率为15.9%，弯曲杆菌DNA为62.2%，可培养弯曲杆菌为1.2%。在实验围栏中，24小时后鸡粪中未检测到可培养的弯曲杆菌。令人惊讶的是，“VBNC弯曲杆菌”即使在72小时后仍持续存在。在养殖舍周围环境和自然污染的鸡粪中证实存在“VBNC弯曲杆菌”。实验室研究表明，根据环境条件，VBNC弯曲杆菌在土壤中可保持完整长达28天，在水中至少可保持63天。
