Molecular Mechanisms of Campylobacter Biofilm Formation and Quorum Sensing.

Even though Campylobacter spp. are known to be fastidious organisms, they can survive within the natural environment. One mechanism to withstand unfavourable conditions is the formation of biofilms, a multicellular structure composed of different bacterial and other microbial species which are embedded in an extracellular matrix. High oxygen levels, low substrate concentrations and the presence of external DNA stimulate the biofilm formation by C. jejuni. These external factors trigger internal adaptation processes, e.g. via regulating the expression of genes encoding proteins required for surface structure formation, as well as motility, stress response and antimicrobial resistance. Known genes impacting biofilm formation will be summarized in this review. The formation of biofilms as well as the expression of virulence genes is often regulated in a cell density depending manner by quorum sensing, which is mediated via small signalling molecules termed autoinducers. Even though quorum sensing mechanisms of other bacteria are well understood, knowledge on the role of these mechanisms in C. jejuni biofilm formation is still scarce. The LuxS enzyme involved in generation of autoinducer-2 is present in C. jejuni, but autoinducer receptors have not been identified so far. Phenotypes of C. jejuni strains lacking a functional luxS like reduced growth, motility, oxygen stress tolerance, biofilm formation, adhesion, invasion and colonization are also summarized within this chapter. However, these phenotypes are highly variable in distinct C. jejuni strains and depend on the culture conditions applied.

Staphylococcus aureus ST6-t701 isolates from food-poisoning outbreaks (2006-2013) in Xi'an, China.

The aim of the study was to investigate the molecular epidemiologic of Staphylococcus aureus isolated from seven staphylococcal food-poisoning (SFP) outbreaks between 2006 and 2013 in Xi'an, northwest China. A total of seven S. aureus isolates associated with seven SFP outbreaks were obtained and characterized by determining the multilocus sequence typing, spa typing, pulsed-field gel electrophoresis (PFGE), antimicrobial susceptibility, toxin and resistant genes. The results showed that S. aureus ST6-t701 (71.4%) was the most predominant clone, followed by ST5-t002 and ST59-t172 (each 14.3%). Seven different PFGE patterns were obtained. Moreover, resistance was most frequently observed to trimethoprim and penicillin (each 71.4%), followed by erythromycin (28.6%), ampicillin, clindamycin, and tetracycline (each 14.3%). All strains were susceptible to chloramphenicol, cefoxitin, oxacillin, amikacin, and vancomycin. Three of seven strains displayed resistance to three or more antimicrobials. Resistance genes were found as follows: linA/linA' (100%), blaZ (85.7%), tet(K), ermC, ermT, and ermB (each 14.3%). Other resistant genes were not detected. In addition, the most frequently identified exotoxin genes were seu, lukED, hla, hlb, hld (each 100%), followed by hlg and hlgv (each 85.7%), lukPV (71.4%), sea (57.1%), see (42.9%), etd (28.6%), and seb, sec, sed, sej, and sek (each 14.3%). The results indicated that S. aureus ST6-t701, with a high prevalence in the northwest of China, exhibited multidrug resistance and harbored multiple toxin and resistance genes. Therefore, strict hygienic and preventative measures should be taken in order to avoid the contamination of food by S. aureus and toxin production in foods.

Motility related gene expression of Campylobacter jejuni NCTC 11168 derived from high viscous media.

Flagellation is one of the major virulence factors of Campylobacter jejuni (C. jejuni), enabling bacterial cells to swarm in rather high viscous fluids. The aim of this study was to determine the impact of the surrounding viscosity on the expression of motility related genes of C. jejuni. Therefore, bacterial RNA was extracted from liquid cultures as well as from bacterial cells recovered from the edge and the center of a swarming halo from high viscous media. The expression pattern of selected flagellar and chemotaxis related genes was investigated by RT-PCR. Higher mRNA levels of class 1 and lower levels of class 2 and 3 flagellar assembly genes were detected in cells derived from the edge of a swarming halo than in cells from the center. This indicates different growth states at both locations within the swarming halo. Furthermore, higher mRNA levels for energy taxis and motor complex monomer genes were detected in high viscous media compared to liquid culture, indicating higher demand of energy if C. jejuni cells were cultivated in high viscous media. The impact of the surrounding viscosity should be considered in future studies regarding motility related questions.

The transcriptional response of Arcobacter butzleri to cold shock.

Arcobacter (A.) butzleri is an emerging zoonotic pathogen associated with gastrointestinal diseases, such as abdominal cramps and diarrhea, and is widely detected in animals, showing a high prevalence in poultry and seafood. The survival and adaptation of A. butzleri to cold temperatures remains poorly studied, although it might be of interest for food safety considerations. To address this, growth patterns of eight A. butzleri isolates were determined at 8 °C for 28 days. A. butzleri isolates showed strain-dependent behavior: six isolates were unculturable after day 18, one exhibited declining but detectable cell counts until day 28 and one grew to the stationary phase level. Out of 13 A. butzleri cold shock-related genes homologous to Escherichia coli, 10 were up-regulated in response to a temperature downshift to 8 °C, as demonstrated by reverse transcription-quantitative PCR. Additionally, we compared these data with the cold-shock response in E. coli. Overall, we provide a deeper insight into the environmental adaptation capacities of A. butzleri, which we find shares similarities with the E. coli cold-shock response.

Prevalence and Characterization of Campylobacter jejuni Isolated from Retail Chicken in Tianjin, China.

Campylobacter jejuni is an important foodborne pathogen worldwide; however, there is a lack of information on the prevalence and antibiotic-resistant profile of C. jejuni in the People's Republic of China. We determined the prevalence and characteristics of C. jejuni on the retail level in Tianjin, one of the five national central cities in China. A total of 227 samples of chicken wings, legs, and breasts were collected from supermarkets and wet markets; 42 of these samples were confirmed to be positive for Campylobacter contamination. The contamination rates of C. jejuni and other Campylobacter species were 13.7% (31 of 227 samples) and 5.7% (13 of 227 samples), respectively. A group of 31 C. jejuni isolates was subjected to antimicrobial susceptibility testing. All (100%) the selected isolates were resistant to ciprofloxacin and nalidixic acid; 77.4% were resistant to tetracycline, 67.7% to doxycycline, 35.5% to gentamicin, 25.8% to clindamycin and florfenicol, 19.4% to chloramphenicol, and 12.9% to erythromycin and azithromycin. A remarkably high proportion (41.9%) of multidrug-resistant isolates was identified. Multilocus sequence typing was conducted to study the population structure of the C. jejuni strains and their relationship to human isolates. The correlation between antimicrobial resistance traits and certain sequence types (STs) or clonal complexes was determined as well. A great genetic diversity of poultry isolates was identified, with 11 STs belonging to 6 clonal complexes and 11 singleton STs. The novel STs accounted for 40.9% (n = 9) of the 22 STs. ST-21, ST-353, ST-354, ST-443, ST-607, and ST-828 complexes had been previously identified from human isolates. This study revealed an extensive level of antimicrobial resistance and genetic diversity in C. jejuni isolated from chicken products in Tianjin, highlighting the necessity of performing enforced interventions to reduce Campylobacter prevalence in China.