Culture and Detection of Campylobacter jejuni within Mixed Microbial Populations of Biofilms on Stainless Steel

Abstract

Campylobacter jejuni is the most frequently reported cause of foodborne illness in the United States, but its survival outside the host is poor. The objective of this research was to examine the formation and composition of biofilms by C. jejuni alone and within mixed bacterial populations from the poultry-processing environment. C. jejuni growth was assessed with four media, two temperatures, and two atmospheric conditions to develop culture methods for liquid media that would allow growth within the biofilms. Growth kinetics was followed at four cell densities to determine temporal compatibility within biofilm mixtures. Analysis of the biofilms by confocal laser scanning microscopy showed that C. jejuni formed a biofilm when incubated without other bacteria. The average surface area of stainless steel covered by C. jejuni increased by 50% from 24 to 48 h, remained level to 96 h, and then decreased by 88% by 168 h. C. jejuni and mixed bacterial populations formed biofilms during incubation periods of up to 7 days. The area of the mixture was significantly greater than for C. jejuni alone at 24 h, was approximately the same at 48 h, and was significantly less by 168 h. When incubated with either of two initial inoculum densities of other bacteria, the number of C. jejuni was enhanced after 24 h. The intensity of fluorescence and cell viability were monitored by epifluorescence microscopy. This study provides the basis for studying interactions of Campylobacter spp. with other bacteria in the environment, which will aid in the design of effective intervention strategies.

Does Campylobacter jejuni Form Biofilms in Food-Related Environments?

“…jejuni can form monospecies biofilms when grown in vitro in culture media on a variety of surfaces, including stainless steel (19,25,26,32), glass (4,19,30), nitrocellulose membranes (19), and various plastic surfaces (25,27,28,31,33,34). Not all studies confirm this pathogen's ability to form biofilms on all surfaces, and findings to the contrary are not uncommon.…”

“…Direct imaging of C. jejuni biofilms using confocal laser scanning microscopy (CLSM) or scanning electron microscopy (SEM) have been used in some studies (4,19,26,32,39). These studies have demonstrated that C. jejuni biofilms may occur in three different forms in liquid culture: (i) cells attached to inert surfaces; (ii) flocs (aggregates of cells floating in the liquid); and (iii) pellicles (aggregates of cells formed at the air-liquid interface) (4,19,40).…”

Influência da vida séssil e planctônica no perfil de resistência antimicrobiana de Campylobacter jejuni isoladas de carcaças de frangos comercializadas no Brasil

“…jejuni pode formar três tipos diferentes de biofilmes: a) estrutura anexada a uma superfície abiótica; b) agregados flutuando na cultura líquida; c) uma película formada na interface gás / líquido (JOSHUA et al, 2006). Geralmente, a formação do biofilme ocorre dentro de 48h de cultivo, mas quando o período é prolongado, sua formação pode ser facilitada e ocorrer o desprendimento celular, caracterizando sua maturação (SANDERS et al, 2007;ICA et al, 2012).…”

“…jejuni pode ser encontrada em biofilmes de espécies mistas (SANDERS et al, 2007;ICA et al, 2012). Os biofilmes multicelulares são estruturalmente complexos e oferecem um ambiente protetor para a sobrevivência em habitats hostis.…”

“…Os biofilmes multicelulares são estruturalmente complexos e oferecem um ambiente protetor para a sobrevivência em habitats hostis. Podem também desempenhar papel importante na dispersão de agentes patogênicos (SANDERS et al, 2007;ICA et al, 2012;OTTO, 2014, SOLANO et al, 2014.…”