A Rapid Culture Method for the Detection of Campylobacter from Water Environments

Abstract

The natural environment and water are among the sources of Campylobacter jejuni and Campylobacter coli. A limited number of protocols exist for the isolation of campylobacters in poorly filterable water. Therefore, the goal of our work was to find a more efficient method of Campylobacter isolation and detection from wastewater and surface water than the ISO standard. In the novel rapid culture method presented here, samples are centrifuged at high speed, and the resuspended pellet is inoculated on a filter, which is placed on Campylobacter selective mCCDA agar. The motile bacteria pass through the filter pores, and mCCDA agar suppresses the growth of background microbiota on behalf of campylobacters. This culture-based method is more efficient for the detection and isolation of Campylobacter jejuni and Campylobacter coli from poorly filterable water than the ISO 17995 standard. It also is less time-consuming, taking only 72 h and comprising three steps, while the ISO standard method requires five or six steps and 144–192 h. This novel culture method, based on high-speed centrifugation, bacterial motility, and selective cultivation conditions, can be used for the detection and isolation of various bacteria from water samples.

Waterborne pathogens detection technologies: advances, challenges, and future perspectives

“…Modern culture media replicate bacteria's natural habitats by incorporating specific elements, enabling the cultivation of previously uncultivable bacteria. A relatively rapid culture method (72 h) was proposed by centrifuging samples and using modified charcoal-cefoperazone-deoxycholate agar (suppress the growth of background microbiota) to isolate Campylobacter more efficiently from poorly filterable water than the ISO 17995 standard (144 to 192 h) (Strakova et al, 2021). It also offers the potential for diverse bacterial detection from water samples.…”

Genotyping of Campylobacter jejuni and prediction tools of its antimicrobial resistance

“…Campylobacters can be easily detected in water by non-cultivation methods in easy way, albeit without the possibility of further genotyping, storage or their sensitivity/resistance to antimicrobials (Van Dyke et al 2010 ). Therefore, extensive water sampling from municipal wastewater treatment plants and surface waters, mainly ponds and lakes in different locations, was performed as was in detail described in our previous study (Strakova et al 2021 ). Such strains of C. jejuni that were difficult to obtain from water ( n = 12) were supplemented by strains isolated from soil, air, and animals.…”

“…The number of human and animal C. jejuni strains was approximately equal. The identity of C. jejuni strains isolated from human, environmental, and animal groups was verified by species-specific multiplex PCR and time-of-flight laser desorption/ionization mass spectrometry (MALDI-TOF/MS) as has been described previously (Kolinska et al 2008 ; Strakova et al 2021 ). The strains were stored at − 80 °C, and prior to every experiment, they were resuscitated by 72-h cultivation at 42 °C under a microaerobic atmosphere on mCCDA agar plates (Oxoid, UK).…”

The effect of environmental conditions on the occurrence of Campylobacter jejuni and Campylobacter coli in wastewater and surface waters

“…(International Organization for Standardisation, 2019 ) was used for the detection of thermotolerant Campylobacter spp. with a slight modification (Strakova et al, 2021 ). Briefly, prefilters with a pore size of 1.4 µm (glass filter; Macherey Nagel) were used to remove mechanical particles from water before sample filtration.…”

“…Suspect Campylobacter spp. colonies ( n = 3) on blood agar were identified by multiplex PCR and matrix‐assisted laser desorption/ionisation time of flight mass spectrometry (MALDI‐TOF/MS) (Strakova et al, 2021 ). Briefly, bacterial DNA was extracted by thermal lysis and PCR was performed using the PPP master mix (Top‐Bio) with primers (Generi Biotech) previously described for the detection of C .…”