Status quo: Levels of Campylobacter spp. and hygiene indicators in German slaughterhouses for broiler and turkey.

Poultry is a common reservoir for Campylobacter and a main source for human campylobacteriosis. With broiler being the predominant poultry for food production, most food safety related research is conducted for this species, for turkey, few studies are available. Although animals are typically colonized at the farm level, the slaughtering process is considered an important factor in re- and cross-contamination. We examined the development of Campylobacter, E. coli and total colony counts (TCC) after several processing steps in three broiler and one turkey slaughterhouses. Whole carcass rinsing and neck skin sampling was applied for broilers resulting in 486 samples in total, while 126 neck skin samples were collected for turkeys. A decrease in the loads of the different bacterial groups along the broiler slaughtering process was observed. Campylobacter mean counts dropped from 4.5 ± 1.7 log10 CFU/ml after killing to 1.6 ± 0.4 log10 CFU/ml after chilling. However, an increase in Campylobacter counts was evident after evisceration before the values again decreased by the final processing step. Although the Campylobacter prevalence in the turkey samples showed a similar development, the bacterial loads were much lower with 1.7 ± 0.3 log10 CFU/g after killing and 1.7 ± 0.4 log10 CFU/g after chilling compared to those of broilers. The loads of E. coli and total colony count of turkey were higher after killing, were reduced by scalding and remained stable until after chilling. This study highlights trends during the slaughtering process in reducing the levels of Campylobacter, E. coli, and total colony counts for broiler and turkey carcasses, from the initial step to after chilling. These results contribute to our understanding of microbial dynamics during meat processing.

Arcobacter butzleri isolates exhibit pathogenic potential in intestinal epithelial cell models.

AIMS: The pathogenic potential of Arcobacter butzleri isolates on human (HT-29/B6) and porcine epithelial (IPEC-J2) cells was investigated by in vitro assays. METHODS AND RESULTS: Five of six A. butzleri isolates were able to adhere and invade HT-29/B6 cells while only four isolates adhered and two invaded IPEC-J2 cells. Two non- or poorly invasive A. butzleri isolates were highly cytotoxic to differentiated HT-29/B6 cells but none to IPEC-J2 cells as determined by WST-assays. Epithelial integrity of cell monolayers, monitored by measurement of the transepithelial electrical resistance (TER), was decreased by all A. butzleri isolates in HT-29/B6 and IPEC-J2 cells to 30-15% and 90-50% respectively. CONCLUSION: The A. butzleri strain-specific pathomechanisms observed with the human colon cell line HT-29/B6, like adhesion, invasion and cytotoxicity might all contribute to epithelial barrier dysfunction, which could explain a leak-flux type of diarrhoea in humans. In contrast, porcine cells seem to be less susceptible to A. butzleri. SIGNIFICANCE AND IMPACT OF THE STUDY: Arcobacter butzleri has enteric pathogenic potential, characterized by defined interactions with human epithelial cells and strain-specific pathomechanisms.

Loads and antimicrobial resistance of Campylobacter spp. on fresh chicken meat in Nueva Ecija, Philippines.

This study was performed to determine the prevalence and to semiquantify Campylobacter spp. on chicken meat samples at 4 selected local wet markets in Nueva Ecija, Philippines, and to determine the antimicrobial resistance patterns of the Campylobacter isolates. Out of 120 chicken meat samples, 57 (47.5%) were Campylobacter spp. positive. The majority of isolated Campylobacter strains were identified as Campylobacter coli (54.4%) and 45.6% as Campylobacter jejuni. Most of these positive samples (52.6%) showed a very high quantitative Campylobacter contamination (most probable number > 2,400/g, lower confidence limit 580/g). For antimicrobial resistance testing, 44 C. coli/jejuni isolates were tested using the agar disk diffusion method. Out of these, 77.3% were resistant to ampicillin, followed by ciprofloxacin (70.4%), tetracycline (54.6%), erythromycin (20.2%), and gentamicin (11.4%). Of the isolates, 36.4% (n = 16) were resistant to 1 antimicrobial agent, 34.1% (n = 15) were resistance to 3 antimicrobial agents, 13.6% (n = 6) to 2 antimicrobial agents, 9.1% (n = 4) to 4 antimicrobial agents, and 6.8% (n = 3) to all 5 antimicrobial agents tested. Our data demonstrate a high contamination of fresh chicken meat with Campylobacter spp. at retail in the Philippines. The detected high Campylobacter prevalences and quantitative loads on chicken meat at retail in the Philippines highlight the need to implement efficient intervention measures along the food chain and to encourage sanitary handling of poultry meat.

Inhibitory effect of high-dosage zinc oxide dietary supplementation on Campylobacter coli excretion in weaned piglets.

AIMS: This study investigated the impact of zinc oxide (ZnO) on Campylobacter coli by in vivo and in vitro assays. METHODS AND RESULTS: By in vitro growth inhibition assays, a high susceptibility of Camp. coli against ZnO could be observed. At concentrations ≥ 2.6 mmol l(-1) ZnO, a decline in cell numbers occurred. Quantitative real-time PCR assays demonstrated an up-regulation of the main oxidative stress gene (katA) in response to ZnO treatment. The expression level of katA was increased by fivefold after ZnO treatment. An experiment was carried out in pigs to elucidate the impact of ZnO as feed supplement on Camp. coli faecal excretion. Feeding a high-dosage ZnO concentration (3100 mg kg(-1) ) to piglets significantly reduced the faecal excretion of Camp. coli by up to 1 log CFU g(-1) as compared to animals receiving a low (40 mg kg(-1) ) or medium (100 mg kg(-1) ) ZnO diet. CONCLUSION: In vitro assays showed a high susceptibility of Camp. coli against ZnO. Adding high levels of ZnO to the diet of weaned piglets reduced Camp. coli excretion significantly. There is evidence for the induction of an oxidative stress response by ZnO supplementation in Camp. coli. SIGNIFICANCE AND IMPACT OF THE STUDY: Supplementation of a high-dosage ZnO diet to piglets can reduce the Camp. coli load, potentially leading to a lower contamination risk of meat during slaughter.

Presence of virulence genes, adhesion and invasion of Arcobacter butzleri.

AIMS: The pathogenic potential of Arcobacter butzleri isolates was investigated by detecting the presence of putative virulence genes and analysing the adhesive and invasive capabilities in cell cultures of human cell lines. METHODS AND RESULTS: The presence of ten putative virulence genes in 52 A. butzleri isolates was determined by PCR. The genes ciaB, mviN, pldA, tlyA, cj1349 and cadF were detected in all, whilst irgA (15%), iroE (60%), hecB (44%) and hecA (13%) were detected only in few A. butzleri isolates. On HT-29 cells, four of six isolates adhered to and three of them were able to invade, whilst all six isolates adhered to and invaded Caco-2 cells with higher degrees. The genes ciaB, cadF and cj1349 of all six isolates were sequenced, but no considerable changes of the amino acids in putative functional domains were observed. CONCLUSION: Selected A. butzleri isolates adhere to and invade HT-29 and Caco-2 cells, which emphasize their human pathogenic potential. The efficiency of invasion depends on the eukaryotic cell line and individual bacterial strain used. We could not show any functional correlation between the amino acid sequence of CadF, CiaB or Cj1349 and the adhesive and invasive phenotype. SIGNIFICANCE AND IMPACT OF THE STUDY: We have shown that some A. butzleri strains invade various cell lines. This underlines their pathogenic potential and hints at their relevance in human disease.

Prevalence, quantitative load, and antimicrobial resistance of Campylobacter spp. from broiler ceca and broiler skin samples in Thailand.

This study was conducted to determine the prevalence of Campylobacter spp. in broiler flocks by testing cecal contents at slaughter and to detect and quantify Campylobacter on broiler carcass skin samples of the corresponding slaughter batches, to determine antimicrobial resistance patterns of the Campylobacter isolates, and to genotype selected Campylobacter jejuni isolates using multilocus sequence typing analysis. Ninety-eight broiler flocks were included in the study. Intact ceca were randomly taken at the time of evisceration throughout a slaughter batch to detect Campylobacter spp. at the broiler flock level and one whole carcass per slaughter batch was taken for the detection of Campylobacter spp. on broiler skin. The prevalences of Campylobacter spp. in broiler ceca and broiler skin samples were 11.2% (11/98) and 51% (50/98), respectively. Even though most Campylobacter-positive broiler skin samples were contaminated with only up to 230 most probable number per gram, a substantial share (13.3%) showed very high Campylobacter numbers on the broiler skin samples (most probable number = ∞; lower confidence limit T(0) 580/g). From 32 C. jejuni and Campylobacter coli isolates tested, the highest antimicrobial resistance rates were found for ciprofloxacin (81.2%), followed by tetracycline (40.6%), ampicillin (31.2%), and erythromycin (9.4%). All tested strains were sensitive to gentamicin. By multilocus sequence typing analysis, a total of 9 different sequence types were identified among 16 C. jejuni isolates. Campylobacter jejuni isolated from cecal content and carcass skin of the same farm or slaughter batch showed corresponding allelic profiles. Our data suggest that intense cross-contamination during the slaughter process led to a strong increase of Campylobacter prevalence on broiler skin compared with the prevalence in broiler ceca. To reduce Campylobacter prevalences on broiler skin, on-farm biosecurity measures need to be accompanied by control measures at the slaughterhouse to reduce fecal contamination of broiler skin and to minimize cross-contamination.

LuxS distribution and AI-2 activity of Campylobacter spp.

AIMS: This study investigates the distribution of LuxS within Campylobacter (Camp.) species and Autoinducer (AI)-2 activity of Camp. jejuni NCTC 11168 in food matrices. METHODS AND RESULTS: LuxS (S-ribosylhomocysteinase) sequences of different Campylobacter spp. were compared, and AI-2 activity was measured with an AI-2 reporter assay. Highest LuxS homologies were shared by Camp. jejuni, Camp. coli and Camp. upsaliensis, and their LuxS sequences had more similarities to the analysed Arcobacter and Vibrio harveyi strains than to all other analysed Campylobacter species. Of 15 analysed species only Camp. lari, Camp. peloridis and Camp. insulaenigrae did not produce AI-2 molecules. Cultivation of Camp. jejuni NCTC 11168 in chicken juice reduced AI-2 activity, and this reduction is not because of lower luxS expression or functionality. CONCLUSION: Not all Campylobacter species encode luxS. Food matrices can reduce AI-2 activity in a LuxS-independent manner. SIGNIFICANCE AND IMPACT OF THE STUDY: Besides, Camp. lari, Camp. peloridis and Camp. insulaenigrae do not show AI-2 activity. Further investigations should clarify the function of AI-2 in Campylobacter spp. and how species lacking luxS could overcome this alteration. Furthermore, the impact of food matrices on these functions needs to be determined as we could show that chicken juice reduced AI-2 activity.

Quorum sensing dependent phenotypes and their molecular mechanisms in Campylobacterales.

Quorum sensing comprises the mechanism of communication between numerous bacteria via small signalling molecules, termed autoinducers (AI). Using quorum sensing, bacteria can regulate the expression of multiple genes involved in virulence, toxin production, motility, chemotaxis and biofilm formation, thus contributing to adaptation as well as colonisation. The current understanding of the role of quorum sensing in the lifecycle of Campylobacterales is still incomplete. Campylobacterales belong to the class of Epsilonproteobacteria representing a physiologically and ecologically diverse group of bacteria that are rather distinct from the more commonly studied Proteobacteria, such as Escherichia and Salmonella. This review summarises the recent knowledge on distribution and production of AI molecules, as well as possible quorum sensing dependent regulation in the mostly investigated species within the Campylobacterales group: Campylobacter jejuni and Helicobacter pylori.

Neural correlates of recognition memory with and without recollection in patients with Alzheimer's disease and healthy controls.

To dissociate recognition memory with and without recollection, event-related potentials (ERPs) of patients with Alzheimer's disease (AD) and matched controls were recorded in a test of verbal recognition memory accompanied by a source judgement. AD patients who had smaller hippocampi showed a disability to recollect the study context (source). Their ERPs elicited by correctly recognized old words compared to new items were more positive only between 300 and 500 ms with a maximum over the frontal scalp. Controls exhibited a sustained old/new effect over left temporoparietal and frontal sites. The present findings suggest that preserved recognition memory in patients with mild AD is independent of hippocampally mediated processes recollecting episodic memories.