Goetz Coralie, Tremblay Yannick D N, Lamarche Daphnée, Blondeau Andréanne, Gaudreau Annie M, Labrie Josée, Malouin François, Jacques Mario
Department of Pathology and Microbiology, Faculty of Veterinary Medicine, Université de Montréal, St-Hyacinthe, Québec, Canada J2S 2M2; Canadian Bovine Mastitis and Milk Quality Research Network, St-Hyacinthe, Québec, Canada J2S 2M2.
Department of Pathology and Microbiology, Faculty of Veterinary Medicine, Université de Montréal, St-Hyacinthe, Québec, Canada J2S 2M2.
J Dairy Sci. 2017 Aug;100(8):6454-6464. doi: 10.3168/jds.2017-12629. Epub 2017 Jun 16.
Coagulase-negative staphylococci (CNS) are considered to be commensal bacteria in humans and animals, but are now also recognized as etiological agents in several infections, including bovine mastitis. Biofilm formation appears to be an important factor in CNS pathogenicity. Furthermore, some researchers have proposed that CNS colonization of the intramammary environment has a protective effect against other pathogens. The mechanisms behind the protective effect of CNS have yet to be characterized. The aim of this study was to evaluate the effect of CNS isolates with a weak-biofilm phenotype on the biofilm formation of other staphylococcal isolates. We selected 10 CNS with a weak-biofilm phenotype and 30 staphylococcal isolates with a strong-biofilm phenotype for this study. We measured biofilm production by individual isolates using a standard polystyrene microtiter plate assay and compared the findings with biofilm produced in mixed cultures. We confirmed the results using confocal microscopy and a microfluidic system with low shear force. Four of the CNS isolates with a weak-biofilm phenotype (Staphylococcus chromogenes C and E and Staphylococcus simulans F and H) significantly reduced biofilm formation in approximately 80% of the staphylococcal species tested, including coagulase-positive Staphylococcus aureus. The 4 Staph. chromogenes and Staph. simulans isolates were also able to disperse pre-established biofilms, but to a lesser extent. We also performed a deferred antagonism assay and recorded the number of colony-forming units in the mixed-biofilm assays on differential or selective agar plates. Overall, CNS with a weak-biofilm phenotype did not inhibit the growth of isolates with a strong-biofilm phenotype. These results suggest that some CNS isolates can negatively affect the ability of other staphylococcal isolates and species to form biofilms via a mechanism that does not involve growth inhibition.
凝固酶阴性葡萄球菌(CNS)被认为是人和动物体内的共生菌,但现在也被确认为包括牛乳腺炎在内的多种感染的病原体。生物膜形成似乎是CNS致病性的一个重要因素。此外,一些研究人员提出,乳腺内环境中的CNS定殖对其他病原体具有保护作用。CNS保护作用背后的机制尚未明确。本研究的目的是评估具有弱生物膜表型的CNS分离株对其他葡萄球菌分离株生物膜形成的影响。我们选择了10株具有弱生物膜表型的CNS和30株具有强生物膜表型的葡萄球菌分离株进行本研究。我们使用标准聚苯乙烯微量滴定板分析法测量单个分离株的生物膜产量,并将结果与混合培养物中产生的生物膜进行比较。我们使用共聚焦显微镜和低剪切力微流控系统证实了结果。4株具有弱生物膜表型的CNS分离株(产色葡萄球菌C和E以及模仿葡萄球菌F和H)显著降低了约80%受试葡萄球菌种类的生物膜形成,包括凝固酶阳性的金黄色葡萄球菌。这4株产色葡萄球菌和模仿葡萄球菌分离株也能够分散预先形成的生物膜,但程度较小。我们还进行了延迟拮抗试验,并在鉴别或选择性琼脂平板上的混合生物膜试验中记录了菌落形成单位的数量。总体而言,具有弱生物膜表型的CNS并未抑制具有强生物膜表型的分离株的生长。这些结果表明,一些CNS分离株可通过一种不涉及生长抑制的机制对其他葡萄球菌分离株和种类形成生物膜的能力产生负面影响。