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.

Development of a new antimicrobial concept for boar semen preservation based on bacteriocins.

The conventional storage temperature of 16-18 °C provides optimal conditions for the preservation of boar sperm quality, which are extremely cold sensitive cells. On the other hand, however, it requires the addition of antibiotics to inhibit bacterial growth. Rising numbers of antibiotic resistant bacteria call for alternatives to this conventional storing method. As potential alternative, three different bacteriocin candidates with known bacteriolytic activity against E. coli were examined on possible negative effects concerning the sperm quality and on their impact on bacterial growth of E. coli ILSH 02692 in BTS-extended semen w/o antibiotics. Although the lower concentrations (0.01 and 0.25%) of all bacteriocins did not show any impact on the quality of the semen, the higher concentrations (0.5 and 1.0%) of two bacteriocins led to a significant (P < 0.05) reduction in several sperm quality characteristics. The bacteriocin 860/1c after AMS/dialysis did not affect the sperm quality in any of the tested concentrations and in all tested extenders (BTS, MIII, Androstar Premium and Androhep all w/o antibiotics) at 16 °C as well as at 6 °C. This bacteriocin reduced growth of E. coli ILSH 02692 in BTS-extended semen by 50% compared to the control w/o bacteriocin. Furthermore, a preliminary insemination trial indicated no impact of the selected bacteriocin on fertility. These promising results show that the application of bacteriocins in liquid-preserved semen is a feasible possibility in the future.

In vivo efficacy of carvacrol on Campylobacter jejuni prevalence in broiler chickens during an entire fattening period.

Carvacrol, a primary constituent of plant essential oils (EOs), and its antimicrobial activity have been the subject of many in vitro studies. Due to an increasing demand for alternative antimicrobials and an emerging number of antibiotic resistant bacteria, the use of essential oils has played a major role in many recent approaches to reduce Campylobacter colonization in poultry before slaughter age. For that purpose, the reducing effect of carvacrol on Campylobacter jejuni prevalence in broilers was determined in vivo in an experimental broiler chicken model during an entire fattening period. Carvacrol was added to the feed in a concentration of 120 mg/kg feed four days post hatch until the end of the trial. In this study, we demonstrated a statistically significant decrease of C. jejuni counts by 1.17 decadic logarithm (log10) most probable number (MPN)/g in cloacal swabs during starter and grower periods (corresponding to a broilers age between 1 and 28 days). Similar results were observed for colon enumeration at the end of the trial where C. jejuni counts were significantly reduced by 1.25 log10 MPN/g. However, carvacrol did not successfully reduce Campylobacter cecal colonization in 33-day-old broilers.

Comparison of consumer knowledge about Campylobacter, Salmonella and Toxoplasma and their transmissibility via meat: results of a consumer study in Germany.

BACKGROUND: Campylobacter is the most commonly reported causative agent of foodborne bacterial infection in Germany, and contaminated chicken meat is an important source of this zoonotic agent. The aim of this study was to determine the knowledge of consumers in Germany about Campylobacter, Salmonella and Toxoplasma and their transmissibility via meat. In addition, we investigated the level of knowledge between selected consumer groups and whether the results coincided with those of international studies. METHODS: We conducted a cross-sectional survey of 1008 consumers in Germany via an online panel to record, analyse and evaluate the state of knowledge about Campylobacter, Salmonella and Toxoplasma. The participants were selected according to age, gender and federal states to be representative of the German population. RESULTS: Overall, 68.3% of the respondents had never heard of Campylobacter, 20.2% had heard of Campylobacter but did not know how to protect themselves, and only 11.5% knew how to protect themselves from Campylobacter infections. Slightly more than half (52.2%) of the respondents who had at least heard of Campylobacter knew that Campylobacter was transmissible via meat. Knowledge increased significantly with age. Participants over 60 years old knew about Campylobacter almost three times as often as the 16- to 19-year-old comparison group (OR = 2.982). Consumers who had at least a secondary school certificate were almost twice as likely to know about Campylobacter as those who had no school certificate or a lower secondary school certificate (OR = 1.899). Participants who were not actors in the food chain were significantly less frequently informed about Campylobacter than were those who were actors in the food chain. Consumer knowledge of Toxoplasma was better than that of Campylobacter. Consumers have the most knowledge about Salmonella. CONCLUSIONS: Consumers in Germany are predominantly poorly informed about Campylobacter and the transmission route via meat. General knowledge of Toxoplasma is better than that of Campylobacter. Among the three pathogens, consumers are best informed about Salmonella. This finding highlights the importance of making existing information materials more accessible to consumers in the future to increase their knowledge, with the objective of reducing the incidence of Campylobacter infections.

Inactivation of Vibrio sp. in pure cultures and mussel homogenates using high hydrostatic pressure.

The objective of this study was to determine the effect of high hydrostatic pressure (HHP) on the inactivation of Vibrio sp. in pure cultures and mussel homogenates. Four Vibrio strains including V. alginolyticus, V. cholerae, V. parahaemolyticus and V. vulnificus were used. HHP treatments were performed with both pure Vibrio sp. cultures in alkaline peptone water (2% NaCl) and artificially inoculated mussel homogenates at pressure levels of 250, 350 and 450 MPa for 1 and 3 min at 25°C. Counts of Vibrio were determined before and after treatment using drop plating method. The effect of high pressure on the reduction level significantly differed among the respective Vibrio species. Vibrio vulnificus was the most susceptible species to HHP. To achieve a >5 log reduction in mussel homogenates, pressure treatment needs to be (i) 350-450 MPa for ≥1 min at 25°C for both V. alginolyticus and V. cholerae, (ii) 250 MPa for ≥3 min or 350-450 MPa for ≥1 min for V. vulnificus and (iii) 350 MPa for ≥3 min or 450 MPa for ≥1 min for V. parahaemolyticus. SIGNIFICANCE AND IMPACT OF THE STUDY: High hydrostatic pressure (HHP) has been applied to inactivate spoilage and pathogenic micro-organisms in a variety of food products, including seafood. Vibrio sp. are frequently reported as the main cause of foodborne illness associated with consumption of raw or undercooked seafood particularly shellfish worldwide. To date, data on the inactivation of Vibrio sp. via HHP are still limited and most of the trials only investigated HHP application in oysters and clams. This study demonstrates the efficacy of HHP inactivating Vibrio sp. in both pure culture and mussel homogenates.

[Epidemiological and hygiene assessment of the health status of population in the area of the construction site of low-head water power development].

Hydrotechnical facilities have significant environmental and health imyacts. Results of a hygienic assessment of the condition of environmental objects in the area of the yrojected Nizhny Novgorod low-head water ylant develoyment showed that the both air and water pollution level in decentralized water supply sources in Balakhna and Gorodets are lower than in the city of Nizhny Novgorod, and those of the water in the water supply network are higher than in the reference area. A cross-sectional study has revealed in the main grouf (Gorodets, Zavolzhye and Balakhna) the rate of morbidity of nervous system and cardiovascular diseases to be higher than in the reference group (Nizhny Novgorod), with cardiovascular diseases being the largest contributor to the overall grevalence rate in the groug under analysis; and digestive system diseases, in the reference group. The study has no revealed direct correlation between morbidity rate in children and adults and the studied factors. However, a direct relationship has been found between the prevalence rate of endocrine diseases in children and the quality of water in the water supply network in Balakhna District (p<0.007); also, there's a correlation between the morbidity rate of digestive diseases and air pollution and water quality in the water supply network (p<0.009). In the Gorodets District, the same situation has been revealed: the morbidity rate of endocrine diseases in children depends on quality of water from the water supply network (p<0.049); the prevalence rate of digestive diseases, on air pollution and water quality in the water supply network (p<0.048). In the same region in adults the prevalence rate of the nervous system diseases, digestive diseases and diseases of the skin and subcutaneous tissue depends on air pollution and pollution of water in the water supply network (p<0.011, p<0.01 and p<0.02, respectively).

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.

Prevalence and Antimicrobial Resistance of Vibrio spp. in Retail and Farm Shrimps in Ecuador.

The aim of this study was to investigate the prevalence of Vibrio spp. in shrimp at retail and in shrimp farms in Ecuador and to determine the antimicrobial agent resistance patterns of farm isolates. The presence of genes linked to early mortality syndrome (EMS) or acute hepatopancreatic necrosis disease (AHPND) also was evaluated. Vibrio spp. were isolated from retail shrimps in Cuenca, Ecuador, and farm shrimps originating from provinces El Oro and Guayas, Ecuador. A total of 229 shrimp samples were collected, of which 71 originated from retail markets in Cuenca and 158 came from shrimp farms. Overall, 219 (95.6%) samples tested positive for Vibrio spp. Vibrio parahaemolyticus (80.8%) was the most common species detected, followed by Vibrio alginolyticus (50.2%), Vibrio cholerae (11.3%), and Vibrio vulnificus (3.5%). None of the V. parahaemolyticus isolates carried the virulence-associated tdh and trh genes. In V. parahaemolyticus shrimp farm isolates, high resistance was found to ampicillin (92.2%), and intermediate resistance was found to tetracycline (51.3%) and amikacin (22.1%). Of the V. parahaemolyticus strains, 68 were resistant to at least three antimicrobial agents, and 2 were resistant to seven antimicrobial agents simultaneously. Up to 18 resistant isolates were found for V. alginolyticus, whereas V. vulnificus and V. cholerae isolates were more susceptible. None of the V. parahaemolyticus isolates carried the EMS-AHPND plasmid. The results of this study revealed the ubiquitous occurrence of Vibrio spp. in shrimps at retail and on shrimp farms in Ecuador.

Gene expression profiles of Vibrio parahaemolyticus in the early stationary phase.

UNLABELLED: Vibrio (V.) parahaemolyticus is an aquatic bacterium capable of causing foodborne gastroenteritis. In the environment or the food chain, V. parahaemolyticus cells are usually forced into the stationary phase, the common phase for bacterial survival in the environment. So far, little is known about whole genomic expression of V. parahaemolyticus in the early stationary phase compared with the exponential growth phase. We performed whole transcriptomic profiling of V. parahaemolyticus cells in both phases (exponential and early stationary phase). Our data showed in total that 172 genes were induced in early stationary phase, while 61 genes were repressed in early stationary phase compared with the exponential phase. Three functional categories showed stable gene expression in the early stationary phase. Eleven functional categories showed that up-regulation of genes was dominant over down-regulation in the early stationary phase. Although genes related to endogenous metabolism were repressed in the early stationary phase, massive regulation of gene expression occurred in the early stationary phase, indicating the expressed gene set of V. parahaemolyticus in the early stationary phase impacts environmental survival. SIGNIFICANCE AND IMPACT OF THE STUDY: Vibrio (V.) parahaemolyticus is one of the main bacterial causes of foodborne intestinal infections. This bacterium usually is forced into stationary phase in the environment, which includes, e.g. seafood. When bacteria are in stationary phase, physiological changes can lead to a resistance to many stresses, including physical and chemical challenges during food processing. To the best of our knowledge, highlighting the whole genome expression changes in the early stationary phase compared with exponential phase, as well as the investigation of physiological changes of V. parahaemolyticus such as the survival mechanism in the stationary phase has been the very first study in this field.

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.

[Vibrio infections from food and sea water. Introducing the "VibrioNet"]. / Vibrio-Infektionen durch Lebensmittel und Meerwasser. Das Netzwerk "VibrioNet" stellt sich vor.

Vibrio is a genus of bacteria present in surface and coastal waters as well as in marine organisms worldwide. In many countries, pathogenic Vibrio species are a main cause of bacterial diarrhea, which may result from comsumption of contaminated seafood and fish products or from drinking contaminated water. Vibrio infections may also gain in importance in our regions due to global warming and the increase in the world trade of seafood. The research network "VibrioNet" studies pathogenic Vibrios in the marine environment and in seafood consumed by humans as a potential, new emerging zoonotic agent. An assessment of the risk arising from pathogenic non-cholera-vibrios in central Europe is the target of a multidisciplinary research effort. The research network will be strengthened by cooperations with international partners from countries in which Vibrio infections play a major role (Bangladesh, Chile, India, Thailand, and Vietnam).

Distribution of Campylobacter jejuni isolates from turkey farms and different stages at slaughter using pulsed-field gel electrophoresis and flaA-short variable region sequencing.

The aim of this study was to assess the diversity of thermotolerant Campylobacter spp. isolated from turkey flocks at six rearing farms 1-2 weeks prior to slaughter (360 faecal swab samples) and from 11 different stages at the slaughterhouse (636 caecal, environmental, neck skin and meat samples). A total of 121 Campylobacter isolates were identified to species level using a multiplex PCR assay and were typed by pulsed-field gel electrophoresis (PFGE) and flaA-short variable region (SVR) sequencing. All Campylobacter isolates were identified as Campylobacter jejuni. PFGE analysis with KpnI restriction enzyme resulted in 11 PFGE types (I-XI) and flaA SVR typing yielded in nine flaA-SVR alleles. The Campylobacter-positive turkey flocks A, C and E were colonized by a limited number of Campylobacter clones at the farm and slaughter. The present study confirms the traceability of flock-specific strains (PFGE types I, V and IX; flaA types 21, 36 and 161) from the farm along the entire processing line to meat cuts. It seems that stress factors such as high temperature of the defeathering water (54-56 °C), drying of the carcass skin during air chilling (24 h at 2 °C), and oxygen in the air could not eliminate Campylobacter completely. Campylobacter-negative flocks became contaminated during processing by the same subtypes of Campylobacter introduced into the slaughter house by preceeding positive flocks even if they were slaughtered on subsequent days. Proper and efficient cleaning and disinfection of slaughter and processing premises are needed to avoid cross-contamination, especially in countries with a low prevalence of Campylobacter spp. The majority of flaA SVR alleles displayed a distinct association with a specific PFGE type. However, a linear relationship for all strains among both typing methods could not be established. To specify genetic relatedness of strains, a combination of different genotyping methods, is needed.

[Campylobacteriosis of man : livestock as reservoir for Campylobacter species]. / Die Campylobacteriose des Menschen : Nutztiere als Reservoir für Campylobacter-Arten.

Over the last few years, infections with Campylobacter have significantly increased in Europe and Germany and these bacteria have even surpassed Salmonella as the most prevalent bacteria, causing gastroenteritis. Especially contamination during the handling and consumption of meat products seems to be the most important risk factor which plays a prominent role for transmission to man. In addition, contact with pets and other animals, drinking raw or improperly pasteurized milk, and the tenacity of Campylobacter in different environments, especially water, have also to be considered for an adequate risk assessment. Besides gastroenteritis, arthralgia, and Guillain-Barré syndrome are important clinical complications of Campylobacter infections in man. At the same time, it is mostly unclear why the course of infection in man and in reservoir animals differs significantly, especially as only a few classical bacterial virulence factors have been identified so far. For these reasons, the development of efficient prevention strategies is of utmost importance in order to control campylobacteriosis.

Wines as possible meat marinade ingredients possess antimicrobial potential against Campylobacter.

This research studied the survival of high (7 log cfu/mL) and low (3 log cfu/mL) inoculum levels of Campylobacter in white and red wines and in grape and tomato juices, which could function as potential antimicrobial marinade ingredients. For comparison, survival was also studied in a commercial poultry meat marinade. White and red wines were shown to have very high bactericidal effects against Campylobacter. High counts were rapidly inactivated to undetectable numbers within 15 min in white wine and within 1 h in red wine, and low counts within 15 min in white wine and within 30 min in red wine. By contrast, grape and tomato juices did not possess high bactericidal effects against Campylobacter because even low counts were occasionally detected after 48 h. The commercial marinade had rather high bactericidal effects against Campylobacter; the high counts were inactivated in most cases within 48 h, and all the low counts were inactivated within 3 h. When testing chicken meat inoculated with Campylobacter and subsequently submerged in white or red wine, the antibacterial activity of the wine was largely reduced. Wines lowered the Campylobacter load inoculated on chicken meat by approximately 1 log cfu/mL over 48 h. The results suggest that wines could be used as antimicrobial ingredients together with the addition of further antimicrobial agents in meat marinades to reduce the numbers of Campylobacter in naturally contaminated poultry products, thus lowering the risk of Campylobacter cross-contamination and transmission through food.

Rapid identification and characterization of Vibrio species using whole-cell MALDI-TOF mass spectrometry.

AIMS: Vibrio identification by means of traditional microbiological methods is time consuming because of the many biochemical tests that have to be performed to distinguish closely related species. This work aimed at evaluating the use of MALDI-TOF mass spectrometry for the rapid identification of Vibrio (V.) spp. as an advantageous application to rapidly discriminate the most important Vibrio spp. and distinguish Vibrio spp. from closely related bacterial species like Photobacterium damselae and Grimontia hollisae and other aquatic bacteria like Aeromonas spp. METHODS AND RESULTS: Starting from sub-colony amounts of pure cultures grown on agar plates, a very simple sample preparation procedure was established and combined with a rapid and automated measurement protocol that allowed species identification within minutes. Closely related species like Vibrio alginolyticus and Vibrio parahaemolyticus or Vibrio cholerae and Vibrio mimicus could thus be differentiated by defining signatures of species-identifying biomarker ions (SIBIs). As a reference method for species designation and for determination of relationships between strains with molecular markers, partial rpoB gene sequencing was applied. CONCLUSIONS: The MALDI-TOF MS-based method as well as the rpoB sequence-based approach for Vibrio identification described in this study produced comparable classification results. The construction of phylogenetic trees from MALDI-TOF MS and rpoB sequences revealed a very good congruence of both methods. SIGNIFICANCE AND IMPACT OF THE STUDY: Our results suggest that whole-cell MALDI-TOF MS-based proteometric characterization represents a powerful tool for rapid and accurate classification and identification of Vibrio spp. and related species.