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Campylobacter jejuni causes gastroenteritis in humans and is a major concern in food safety. Commercially prepared chicken meats are frequently contaminated with C. jejuni, which is closely associated with the diffusion of intestinal contents in poultry processing plants. Sodium hypochlorite (NaClO) is commonly used during chicken processing to prevent food poisoning; however, its antimicrobial activity is not effective in the organic-rich solutions. In this study, we investigated the potential of a new photo-disinfection system, UVA-LED, for the disinfection of C. jejuni-contaminated chicken surfaces. The data indicated that UVA irradiation significantly killed C. jejuni and that its killing ability was significantly facilitated in NaClO-treated chickens. Effective inactivation of C. jejuni was achieved using a combination of UVA and NaClO, even in the organic-rich condition. The results of this study show that synergistic disinfection using a combination of UVA and NaClO has potential beneficial effects in chicken processing systems.

Introduction: Campylobacter jejuni is a Gram-negative, microaerophilic bacteria that is the leading cause of food-borne illnesses in the United States; it frequently causes severe diarrhoea. Campylobacter jejuni has recently demonstrated antibiotic resistance; therefore, alternative methods for treating these infections must be investigated. Visible light has proven to be an effective inhibitory agent for many medically significant species of bacteria and fungi., Methods: Visible light in the blue range (405 nm and 464 nm) was administered to a suspension of Campylobacter jejuni. Brain-heart infusion agar was inoculated with the suspension of the organism following exposure to blue light at a dose of 24 Jcm 2 for each wavelength. Inoculated plates were placed into a traditional Campy jar in order to mimic a 5% O 2 atmosphere and incubated at 37 °C for 48 hours. Following incubation, colony forming units were counted and compared with controls to determine if the light energy had effectively inhibited the growth of the organism., Results: Both 405 nm and 464 nm light at a dose of 24 Jcm 2 were shown to be effective inhibitors of Campylobacter jejuni, with kill rates ranging from 82.4-98.9%., Conclusion: Blue light effectively inhibits Campylobacter jejuni., (Copyright © 2019 Elsevier B.V. and International Society of Chemotherapy. All rights reserved.)

Recent outbreaks of Campylobacter mediated disease attributed to undercooked chicken livers have highlighted a continuing need for methods to reduce Campylobacter numbers in these types of food products. In this study, gamma irradiation is evaluated for its effectiveness in reducing Campylobacter jejuni numbers in experimentally contaminated chicken livers. A wide range of radiation doses were evaluated in conjunction with cold storage parameters, before and after irradiation. Storage of chicken livers at -20 °C prior to radiation treatment, as expected, increased C. jejuni radiation resistance. Livers previously stored at -20 °C exhibited D 10 values of 0.748 kiloGray (kGy) compared to livers without previous storage that had a significantly lower D 10 value of 0.361 kGy. Cold storage conditions post-irradiation at both 4 °C and -20 °C further reduced the C. jejuni numbers over those reduced by the initial irradiation. The largest reduction (3.8 logs) of C. jejuni numbers in livers produced by combining irradiation and cold storage was achieved using 0.8 kGy of radiation followed by 1 week storage at -20 °C. This reduction of 3.8 logs was not determined to be significantly different from the 3.5 log reduction achieved with the same radiation dose (0.8 kGy) after only 48 h of subsequent storage at -20 °C., (Published by Elsevier Ltd.)

In this study, a LOV-based fluorescent reporter (light, oxygen, or voltage-sensing domains of phototropin), termed iLOV, was adapted for Campylobacter jejuni and used to investigate promoter activity via monitoring fluorescence intensity and to study the localisation of two chemotaxis proteins. The pC46 complementation vector contains coding sequence from cj0046, a C. jejuni NCTC11168 pseudo-gene and is used to integrate cloned genes onto the C. jejuni chromosome. The pC46 vector was used to construct plasmids containing iLOV, driven by three different C. jejuni constitutive promoters and plasmids containing transcriptional fusions of the iLOV reporter and two chemoreceptors, tlp5 and tlp8. Expression from the porA promoter, pporA, produced the highest fluorescence signals compared to pfdxA (intermediate level) and pmetK (lowest level). The cellular localisation pattern of transducer-like protein (Tlp) clusters, containing Tlp5 and Tlp8, was predominately polar, with Tlp5 positioned only at one and Tlp8 at both poles. Here, we demonstrate that a iLOV fluorescent reporter can be used as a promoter probe or as a gene fusion reporter in Campylobacter spp. This is a new system uniquely placed for studying Campylobacter spp., as it combines resistance to photobleaching and functionality under microaerobic conditions., (Copyright © 2017. Published by Elsevier Masson SAS.)

Campylobacter jejuni, the leading bacterial cause of human gastroenteritis in the United States, displays significant strain diversity due to horizontal gene transfer. Conjugation is an important horizontal gene transfer mechanism contributing to the evolution of bacterial pathogenesis and antimicrobial resistance. It has been observed that heat shock could increase transformation efficiency in some bacteria. In this study, the effect of heat shock on C. jejuni conjugation efficiency and the underlying mechanisms were examined. With a modified Escherichia coli donor strain, different C. jejuni recipient strains displayed significant variation in conjugation efficiency ranging from 6.2 × 10(-8) to 6.0 × 10(-3) CFU per recipient cell. Despite reduced viability, heat shock of standard C. jejuni NCTC 11168 and 81-176 strains (e.g., 48 to 54°C for 30 to 60 min) could dramatically enhance C. jejuni conjugation efficiency up to 1,000-fold. The phenotype of the heat shock-enhanced conjugation in C. jejuni recipient cells could be sustained for at least 9 h. Filtered supernatant from the heat shock-treated C. jejuni cells could not enhance conjugation efficiency, which suggests that the enhanced conjugation efficiency is independent of secreted substances. Mutagenesis analysis indicated that the clustered regularly interspaced short palindromic repeats system and the selected restriction-modification systems (Cj0030/Cj0031, Cj0139/Cj0140, Cj0690c, and HsdR) were dispensable for heat shock-enhanced conjugation in C. jejuni. Taking all results together, this study demonstrated a heat shock-enhanced conjugation efficiency in standard C. jejuni strains, leading to an optimized conjugation protocol for molecular manipulation of this organism. The findings from this study also represent a significant step toward elucidation of the molecular mechanism of conjugative gene transfer in C. jejuni., (Copyright © 2015, American Society for Microbiology. All Rights Reserved.)

Thermotolerant Campylobacter spp. frequently cause bacterial gastroenteritis in humans commonly infected through the consumption of undercooked poultry meat. We examined Campylobacter jejuni heat-stress responses in vitro after exposure to 48°C and 55°C. The in vivo modulation of its pathogenicity was also investigated using BALB/c mice intravenously infected with stressed C. jejuni. Regardless of the bacterial growth phase, the culturability and viability of C. jejuni in vitro was reduced after exposure to 55°C. This correlated with the altered protein profile and decreased virulence properties observed in vivo. Heat stress at 48°C elicited the transition to more resistant bacterial forms, independent of morphological changes or the appearance of shorter spiral and coccoid cells. This treatment did not cause marked changes in bacterial virulence properties in vivo. These results indicated that the characteristics and pathogenicity of C. jejuni in response to heat stress are temperature dependent. Further studies on the responses of C. jejuni to stresses used during food processing, as well as the modulation of its virulence, are important for a better understanding of its contamination and infective cycle, and will, thus, contribute to improved safety in the food production chain.

Control of Campylobacter in the food chain requires a better understanding of the behaviour of the bacteria in relevant environments. Campylobacter species are largely non-pathogenic in poultry, the body temperature of which is 42 °C. However, the bacteria are highly pathogenic in humans whose body temperature is 37 °C. The aim of this study was to examine if switching from commensal to pathogenic behaviour was related to temperature. We examined the growth, motility and invasion of T84 cells by three species of Campylobacter: C. jejuni 81116, C. jejuni M1, C. coli 1669, C. coli RM2228 and C. fetus fetus NC10842 grown at 37 and 42 °C. Our results suggest that C. jejuni isolates grow similarly at both temperatures but some are more motile at 42 °C and some are more invasive at 37 °C, which may account for its rapid spread in poultry flocks and for infection in humans, respectively. C. coli, which are infrequent causes of Campylobacter infections in humans, is less able to grow and move at 37 °C compared to 42 °C but was significantly more invasive at the lower temperature. C. fetus fetus, which is infrequently found in poultry, is less able to grow and invade at 42 °C.

Campylobacter jejuni is a major gastrointestinal pathogen that colonizes host mucosa via interactions with extracellular matrix proteins such as fibronectin. The aim of this work was to study in vitro the adhesive properties of C. jejuni ATCC 33291 and C. jejuni 241 strains, in both culturable and viable but non-culturable (VBNC) forms. To this end, the expression of the outer-membrane protein CadF, which mediates C. jejuni binding to fibronectin, was evaluated. VBNC bacteria were obtained after 46-48 days of incubation in freshwater at 4 °C. In both cellular forms, the expression of the cadF gene, assessed at different time points by RT-PCR, was at high levels until the third week of VBNC induction, while the intensity of the signal declined during the last stage of incubation. CadF protein expression by the two C. jejuni strains was analysed using 2-dimensional electrophoresis and mass spectrometry; the results indicated that the protein, although at low levels, is also present in the VBNC state. Adhesion assays with culturable and VBNC cells, evaluated on Caco-2 monolayers, showed that non-culturable bacteria retain their ability to adhere to intestinal cells, though at a reduced rate. Our results demonstrate that the C. jejuni VBNC population maintains an ability to adhere and this may thus have an important role in the pathogenicity of this microorganism.

Background: The genetic features that facilitate Campylobacter jejuni's adaptation to a wide range of environments are not completely defined. However, whole genome expression studies showed that respiratory proteins (RPs) were differentially expressed under varying conditions and stresses, suggesting further unidentified roles for RPs in C. jejuni's adaptation. Therefore, our objectives were to characterize the contributions of selected RPs to C. jejuni's i- key survival phenotypes under different temperature (37°C vs. 42°C) and oxygen (microaerobic, ambient, and oxygen-limited/anaerobic) conditions and ii- its interactions with intestinal epithelial cells from disparate hosts (human vs. chickens)., Results: C. jejuni mutant strains with individual deletions that targeted five RPs; nitrate reductase (ΔnapA), nitrite reductase (ΔnrfA), formate dehydrogenase (ΔfdhA), hydrogenase (ΔhydB), and methylmenaquinol:fumarate reductase (ΔmfrA) were used in this study. We show that only the ΔfdhA exhibited a decrease in motility; however, incubation at 42°C significantly reduced the deficiency in the ΔfdhA's motility as compared to 37°C. Under all tested conditions, the ΔmfrA showed a decreased susceptibility to hydrogen peroxide (H(2)O(2)), while the ΔnapA and the ΔfdhA showed significantly increased susceptibility to the oxidant as compared to the wildtype. Further, the susceptibility of the ΔnapA to H(2)O(2) was significantly more pronounced at 37°C. The biofilm formation capability of individual RP mutants varied as compared to the wildtype. However, the impact of the deletion of certain RPs affected biofilm formation in a manner that was dependent on temperature and/or oxygen concentration. For example, the ΔmfrA displayed significantly deficient and increased biofilm formation under microaerobic conditions at 37°C and 42°C, respectively. However, under anaerobic conditions, the ΔmfrA was only significantly impaired in biofilm formation at 42°C. Additionally, the RPs mutants showed differential ability for infecting and surviving in human intestinal cell lines (INT-407) and primary chicken intestinal epithelial cells, respectively. Notably, the ΔfdhA and the ΔhydB were deficient in interacting with both cell types, while the ΔmfrA displayed impairments only in adherence to and invasion of INT-407. Scanning electron microscopy showed that the ΔhydB and the ΔfdhA exhibited filamentous and bulging (almost spherical) cell shapes, respectively, which might be indicative of defects in cell division., Conclusions: We conclude that the RPs contribute to C. jejuni's motility, H(2)O(2) resistance, biofilm formation, and in vitro interactions with hosts' intestinal cells. Further, the impact of certain RPs varied in response to incubation temperature and/or oxygen concentration. Therefore, RPs may facilitate the prevalence of C. jejuni in a variety of niches, contributing to the pathogen's remarkable potential for adaptation.

Background: Campylobacter jejuni is a major cause of bacterial food-borne illness in Europe and North America. The mechanisms allowing survival in the environment and transmission to new hosts are not well understood. Environmental free-living protozoa may facilitate both processes. Pre-exposure to heat, starvation, oxidative or osmotic stresses encountered in the environment may affect the subsequent interaction of C. jejuni with free-living protozoa. To test this hypothesis, we examined the impact of environmental stress on expression of virulence-associated genes (ciaB, dnaJ, and htrA) of C. jejuni and on its uptake by and intracellular survival within Acanthamoeba castellanii., Results: Heat, starvation and osmotic stress reduced the survival of C. jejuni significantly, whereas oxidative stress had no effect. Quantitative RT-PCR experiments showed that the transcription of virulence genes was slightly up-regulated under heat and oxidative stresses but down-regulated under starvation and osmotic stresses, the htrA gene showing the largest down-regulation in response to osmotic stress. Pre-exposure of bacteria to low nutrient or osmotic stress reduced bacterial uptake by amoeba, but no effect of heat or oxidative stress was observed. Finally, C. jejuni rapidly lost viability within amoeba cells and pre-exposure to oxidative stress had no significant effect on intracellular survival. However, the numbers of intracellular bacteria recovered 5 h post-gentamicin treatment were lower with starved, heat treated or osmotically stressed bacteria than with control bacteria. Also, while ~1.5 × 103 colony forming unit/ml internalized bacteria could typically be recovered 24 h post-gentamicin treatment with control bacteria, no starved, heat treated or osmotically stressed bacteria could be recovered at this time point. Overall, pre-exposure of C. jejuni to environmental stresses did not promote intracellular survival in A. castellanii., Conclusions: Together, these findings suggest that the stress response in C. jejuni and its interaction with A. castellanii are complex and multifactorial, but that pre-exposure to various stresses does not prime C. jejuni for survival within A. castellanii.

The application of crust freezing (CF) applied as a stand-alone treatment or in combination with ultraviolet (UV) light for reducing the level of artificially inoculated Campylobacter jejuni on raw chicken was investigated. CF air temperatures of -5, -15 and -27 °C (±3 °C) with freezing times of 70, 15 and 6 min, respectively, were used. The level of C. jejuni on chicken was also examined following subsequent refrigerated (0-4 °C) storage at 3 and 7 days. All CF treatments resulted in significant reductions compared to untreated controls (P < 0.05). Although combining CF with UV also resulted in significant reductions for C. jejuni, the combined treatments were generally no more effective than treatment by CF alone. Overall, the color of chicken drumsticks was not affected by CF treatments (P ≥ 0.05). In general, CF resulted in increased drip loss (P < 0.05), which increased over storage time and was greater at higher CF temperatures. The current study indicates that CF has potential for reducing the levels of C. jejuni by between 0.5 and 1.5 log(10) CFU/g and impacts minimally on the color of treated skin., (Copyright © 2012 Elsevier Ltd. All rights reserved.)

Campylobacter is one of the leading causes of human foodborne illnesses originating from meat and poultry products. Cross-contamination of this organism occurs in many poultry processing plants, and can occur in the kitchens and refrigerators of consumers. Therefore, new intervention strategies are needed for meat and poultry products to better protect consumers from this pathogen. Vacuum or modified atmosphere packaging is a common packaging technique used by the meat and poultry industry to extend the shelf life of meat products. In addition, irradiation has been well established as an antibacterial treatment to reduce pathogens on meat and poultry products. Irradiation in combination with high-CO(2) + CO modified atmosphere packaging (MAP) was investigated in this study for the control of Campylobacter jejuni in chicken breast meat. The radiation sensitivity (D(10)-value) of this foodborne pathogen in chicken breast meat was similar in vacuum or high-O(2) MAP (0.31 ± 0.01 kGy in vacuum packaging and 0.29 ± 0.03 kGy in MAP). C. jejuni survived in both vacuum and high-CO(2) MAP through 6 weeks of refrigerated storage. Irradiation was effective for eliminating C. jejuni from meat or poultry packaged in vacuum or MAP, and should reduce the chance of cross-contamination in retail stores or home kitchens. However, irradiated off-odor and sour aroma were observed for raw, irradiated chicken breast packaged with either vacuum or MAP. Therefore, additional means to mitigate quality changes appear necessary for these products.

The chemotactic behaviour of Campylobacter strains was determined in the presence of different amino acids at two temperatures (37 °C and 42 °C). Two strains of catalase positive (Campylobacter jejuni) and negative (Campylobacter sputurum) Campylobacter were isolated from river water in Tonekabon, Iran and identified by phenotyping and 16srRNA Gene sequencing methods. Chemotactic responses of the isolates were assessed toward a variety of amino acids viz., L-cystine, L-asparagine, L-histidine, L-aspartic acid, L-serine, L-phenylalanine, L-leucine and L-tryptophan by disc and capillary methods at two temperatures: 37 °C and 42 °C. C. jejuni showed positive chemotactic response towards L-cystine,L-tryptophan, L-phenylalanine, - L-leucine, L-asparagine and L-Serine at both, 37 °C and 42 °C however, it was greater at 37 °C. C. sputurum showed negative or weak response towards all of the amino acids. In addition, C. jejuni illustrated strong chemotactic response to L-asparagine follow by L-serine and weak chemotaxis response to L-phenylalanine and L-cysteine at 37 °C. Overall, C. jejuni showed relatively strong chemotactic response to some amino acids, likewise it was greater at 37 °C. Hence, the human body temperature (37 °C) in compared to avian body temperature (42 °C) probably promotes chemotactic response of C. jejuni, which it might be a reason for causing disease in human being compared to avian., (Copyright © 2011 Elsevier Ltd. All rights reserved.)

UV light was investigated for the decontamination of raw chicken, associated packaging, and contact surfaces. The UV susceptibilities of a number of Campylobacter isolates (seven Campylobacter jejuni isolates and three Campylobacter coli isolates), Escherichia coli ATCC 25922, and Salmonella enterica serovar Enteritidis ATCC 10376 in liquid media were also investigated. From an initial level of 7 log CFU/ml, no viable Campylobacter cells were detected following exposure to the most intense UV dose (0.192 J/cm(2)) in liquid media (skim milk subjected to ultrahigh-temperature treatment and diluted 1:4 with maximum recovery diluent). Maximum reductions of 4.8 and 6.2 log CFU/ml were achieved for E. coli and serovar Enteritidis, respectively, in liquid media. Considerable differences in susceptibilities were found between the Campylobacter isolates examined, with variations of up to 4 log CFU/ml being observed. UV treatment of raw chicken fillet (0.192 J/cm(2)) reduced C. jejuni, E. coli, serovar Enteritidis, total viable counts, and Enterobacteriaceae by 0.76, 0.98, 1.34, 1.76, and 1.29 log CFU/g, respectively. Following UV treatment of packaging and surface materials, reductions of up to 3.97, 4.50, and 4.20 log CFU/cm(2) were obtained for C. jejuni, E. coli, and serovar Enteritidis, respectively (P < 0.05). Overall, the color of UV-treated chicken was not significantly affected (P ≥ 0.05). The findings of this study indicate that Campylobacter is susceptible to UV technology and that differences in sensitivities exist between investigated isolates. Overall, UV could be used for improving the microbiological quality of raw chicken and for decontaminating associated packaging and surface materials.

The microaerophilic bacterium Campylobacter jejuni is the most common cause of bacterial food-borne infections in the developed world. Tolerance to environmental stress relies on proteases and chaperones in the cell envelope, such as HtrA and SurA. HtrA displays both chaperone and protease activities, but little is known about how each of these activities contributes to stress tolerance in bacteria. In vitro experiments showed temperature-dependent protease and chaperone activities of C. jejuni HtrA. A C. jejuni mutant lacking only the protease activity of HtrA was used to show that the HtrA chaperone activity is sufficient for growth at high temperature or under oxidative stress, whereas the HtrA protease activity is essential only under conditions close to the growth limit for C. jejuni. However, the protease activity was required to prevent induction of the cytoplasmic heat shock response even under optimal growth conditions. Interestingly, the requirement of HtrA at high temperatures was found to depend on the oxygen level, and our data suggest that HtrA may protect oxidatively damaged proteins. Finally, protease activity stimulates HtrA production and oligomer formation, suggesting that a regulatory role depends on the protease activity of HtrA. Studying a microaerophilic organism encoding only two known periplasmic chaperones (HtrA and SurA) revealed an efficient HtrA chaperone activity and proposed multiple roles of the protease activity, increasing our understanding of HtrA in bacterial physiology.

Although considerable research has been carried out on a range of environmental factors that impact on the survival of Campylobacter jejuni, there is limited information on the effects of violet/blue light on this pathogen. This investigation was carried out to determine the effects of high-intensity 405-nm light on C. jejuni and to compare this with the effects on two other important Gram-negative enteric pathogens, Salmonella enteritidis and Escherichia coli O157:H7. High-intensity 405-nm light generated from an array of 405-nm light-emitting diodes was used to inactivate the test bacteria. The results demonstrated that while all three tested species were susceptible to 405-nm light inactivation, C. jejuni was by far the most sensitive organism, requiring a total dose of 18 J cm⁻² of 405-nm light to achieve a 5-log₁₀ reduction. This study has established that C. jejuni is particularly susceptible to violet/blue light at a wavelength of 405 nm. This finding, coupled with the safety-in-use advantages of this visible (non-ultraviolet wavelength) light, suggests that high-intensity 405-nm light may have applications for control of C. jejuni contamination levels in situations where this type of illumination can be effectively applied.

Regulation of the biosynthesis of the flagellar filament in bacteria containing multiple flagellin genes is not well understood. The major food-borne pathogen Campylobacter jejuni possesses on both poles a flagellum that consists of two different flagellin subunits, FlaA and FlaB. Here we identify the protein Cj1464 as a regulator of C. jejuni flagellin biosynthesis. The protein shares characteristics of the FlgM family of anti-sigma factor proteins: it represses transcription of sigma(28)-dependent genes, forms a complex with sigma factor FliA, and is secreted through the flagellar filament. However, unlike other FlgM proteins, the interaction of C. jejuni FlgM with FliA is regulated by temperature and the protein does not inhibit FliA activity during the formation of the hook-basal body complex (HBB). Instead, C. jejuni FlgM limits the length of the flagellar filament by suppressing the synthesis of both the sigma(28)- and the sigma(54)-dependent flagellins. The main function of the C. jejuni FlgM therefore is not to silence sigma(28)-dependent genes until the HBB is completed, but to prevent unlimited elongation of the flagellum, which otherwise leads to reduced bacterial motility.

Campylobacter spp. readily colonize the intestinal tracts of both human and avian species. While most often commensal organisms in birds, campylobacters remain the leading cause of bacterial gastroenteritis in humans. The association of campylobacters with poultry is well established as a primary route for human exposure. The difference in normal core body temperature between chickens (42 degrees C) and humans (37 degrees C) has been suggested to trigger potential colonization or virulence factors and investigators have demonstrated differential gene expression at the two temperatures. Campylobacter spp. exhibit unique nutritional requirements and have been thought to only utilize amino acids and Kreb cycle intermediates as carbon sources for growth. We evaluated the ability of the genome-sequenced strain of Campylobacter jejuni 11168 (GS) to oxidize 190 different substrates as sole carbon sources at 37 degrees C and 42 degrees C using phenotype microarray (PM) technology. Results indicate that the expected amino acids, l-serine, l-aspartic acid, l-asparagine, and l-glutamic acid were utilized in addition to a number of organic acids. In general, oxidation of the substrates was greater at 42 degrees C than at 37 degrees C with a few exceptions. By employing the PM method, we observed a number of potential false-positive reactions for substrates including the triose, dihydroxyacetone; and the pentose sugars, d-xylose, d-ribose, l-lyxose, and d- and l-arabinose. The presence of genes possibly responsible for utilization of pentose sugars is supported by the genomic sequence data, but actual utilization as sole carbon sources for active respiration has not been observed. A better understanding of the metabolic pathways and nutritional requirements of campylobacters could lead to improvements in culture media for detection and isolation of the pathogen and to future intervention methods to reduce human exposure., (Published by Elsevier B.V.)

While maintaining nutritional and sensorial attributes of fresh foods mild processing technologies generally deliver microbiologically perishable food products. Currently little information exists on possible increase in the resistance of pathogens after repetitive exposure to mild (sub-lethal) treatments. Multiple strain-cocktails of Listeria monocytogenes, Escherichia coli O157:H7 and Campylobacter jejuni were exposed to 20 consecutive cycles of sub-lethal inactivation by three different techniques. Used techniques comprised inactivation with lactic acid (LA), chlorine dioxide (ClO(2)) and intense light pulses (ILP). Results showed that the selection of resistant cells was both species and technique dependent. While repetitive cycles of ClO(2) treatment did not result in increased resistance, repetitive inactivation with LA yielded L. monocytogenes culture of higher resistance in comparison to the parental culture. The increased resistance, expressed as decreased level of reduction in bacterial counts in subsequent inactivation cycles, was also observed with ILP for both L. monocytogenes and E. coli O157:H7 strains. Visual trend observations were confirmed through statistical linear regression analysis. No such effects were noted for C. jejuni which became undetectable after first 2-5 cycles. Current findings indicate the ability of foodborne pathogens to adapt to mild bactericidal treatments creating new challenges in risk assessment and more specifically in hazard analysis.

Human infection with Campylobacter jejuni is often associated with the consumption of foods that have been exposed to both chilling and high temperatures. Despite the public health importance of this pathogen, little is known about the effects of cold exposure on its ability to survive a subsequent heat challenge. This work examined the effect of rapid exposure to chilling, as would occur in poultry processing, on the heat resistance at 56 degrees C of two C. jejuni strains, 11168 and 2097e48, and of Escherichia coli K-12. Unlike E. coli K-12, whose cold-exposed cells showed increased sensitivity to 56 degrees C, such exposure had only a marginal effect on subsequent heat resistance in C. jejuni. This may be explained by the finding that during rapid chilling, unlike E. coli cells, C. jejuni cells are unable to alter their fatty acid composition and do not adapt to cold exposure. However, their unaltered fatty acid composition is more suited to survival when cells are exposed to high temperatures. This hypothesis is supported by the fact that in C. jejuni, the ratio of unsaturated to saturated fatty acids was not significantly different after cold exposure, but it was in E. coli. The low-temperature response of C. jejuni is very different from that of other food-borne pathogens, and this may contribute to its tolerance to further heat stresses.

Campylobacter jejuni has the potential to thrive at 37C (e.g., in the human intestinal tract) and 42C (e.g., in the poultry intestinal tract). We aimed to determine the protein expression profiles of C. jejuni cultured at 37C and 42C in vitro by two-dimensional electrophoresis (2DE). The differentially expressed spots/proteins between C. jejuni cultured at 37C and 42C were defined when their expression differed by twofold. The differently expressed spots detected from C. jejuni cultured both on agar and in broth at 37C and 42C were subjected to protein identification by MALDI-TOF/TOF. Overall, 15 and 20 differentially expressed proteins were defined for C. jejuni cultured at the two temperatures on agar and in broth, respectively. All of the identified differentially expressed proteins could be clustered as proteins involving the metabolism, regulator system, periplasmic proteins and the major antigens of C. jejuni. In conclusion, there are subsets of proteins that are optimally expressed at 37C, which may contribute to the host adaptation and/or the pathogenicity in the human intestinal tract.

The presence and functionality of DNA repair mechanisms in Campylobacter jejuni are largely unknown. In silico analysis of the complete translated genome of C. jejuni NCTC 11168 suggests the presence of genes involved in methyl-directed mismatch repair (MMR), nucleotide excision repair, base excision repair (BER), and recombinational repair. To assess the functionality of these putative repair mechanisms in C. jejuni, mutS, uvrB, ung, and recA knockout mutants were constructed and analyzed for their ability to repair spontaneous point mutations, UV irradiation-induced DNA damage, and nicked DNA. Inactivation of the different putative DNA repair genes did not alter the spontaneous mutation frequency. Disruption of the UvrB and RecA orthologues, but not the putative MutS or Ung proteins, resulted in a significant reduction in viability after exposure to UV irradiation. Assays performed with uracil-containing plasmid DNA showed that the putative uracil-DNA glycosylase (Ung) protein, important for initiation of the BER pathway, is also functional in C. jejuni. Inactivation of recA also resulted in a loss of natural transformation. Overall, the data indicate that C. jejuni has multiple functional DNA repair systems that may protect against DNA damage and limit the generation of genetic diversity. On the other hand, the apparent absence of a functional MMR pathway may enhance the frequency of on-and-off switching of phase variable genes typical for C. jejuni and may contribute to the genetic heterogeneity of the C. jejuni population.

The effects of UV irradiation at a wavelength of 254 nm on the survival of Campylobacter jejuni on the surfaces of broiler meat, skin, and carcasses were studied. On broiler carcasses, the effects of UV were also studied in combination with activated oxygen. The surfaces were inoculated with varying counts of C. jejuni and treated with UV irradiation using doses ranging between 9.4 and 32.9 mW/s per square centimeter. The log reductions in C. jejuni counts were determined by dilution plating. The effects of both treatments on the sensory quality of broiler meat, including visual appearance, odor, and fatty acid composition, were also evaluated. On broiler meat, the maximum reduction achieved was 0.7 log and on broiler skin 0.8 log. On broiler carcasses, the maximum reduction using UV irradiation was 0.4 log, and using UV in combination with activated oxygen 0.4 log. No significant differences were found in the sensory quality between the samples and the controls. The use of UV irradiation alone or in combination with activated oxygen cannot be recommended as a primary decontamination method for C. jejuni on broiler carcasses. The use of these methods in combination with other decontamination techniques, and processing with proper processing plant sanitation and hygiene, might be more effective in reducing the C. jejuni counts on broiler carcass surfaces than the use of these methods only.

The inactivation of Campylobacter jejuni and Salmonella enterica, compared with Escherichia coli, was determined in 100 l chambers of seawater and river water located at an outdoor site. The chambers (paired with dark controls) were seeded with waste stabilization pond effluent and laboratory-cultured pathogens, and exposed to sunlight in summer and winter experiments. All sunlight inactivation (k(S)) rates, as a function of cumulative global solar radiation (insolation), were far higher than the corresponding dark (k(D)) rates, with a ranking (and average k(S) rates for seawater and river water, respectively) of: C. jejuni (3.23; 2.34)>S. enterica (0.51; 0.37)>E. coli (0.34; 0.26). All the T(90) (time to 90% inactivation) values were higher in winter than in summer, but there was far greater similarity between the summer and winter S(90) (insolation needed for 90% inactivation) values. The rapid inactivation of C. jejuni was attributed to a high susceptibility to photooxidative damage. The results suggest that, in sunlight-exposed waters, E. coli will be a more conservative indicator for C. jejuni than for S. enterica, and C. jejuni transmission as a pathogenic agent is less likely than for S. enterica.

Raw ground beef patties inoculated with stationary-phase cells of Escherichia coli O157:H7, salmonellae, or Campylobacter jejuni were subjected to gamma irradiation (60Co) treatment, with doses ranging from 0 to 2.52 kGy. The influence of two levels of fat (8 to 14% [low fat] and 27 to 28% [high fat]) and temperature (frozen [-17 to -15 degrees C] and refrigerated [3 to 5 degrees C]) on the inactivation of each pathogen by irradiation was investigated. In ascending order of irradiation resistance, the D10 values ranged from 0.175 to 0.235 kGy (C. jejuni), from 0.241 to 0.307 kGy (E. coli O157:H7), and from 0.618 to 0.800 kGy (salmonellae). Statistical analysis revealed that E. coli O157:H7 had a significantly (P < 0.05) higher D10 value when irradiated at -17 to -15 degrees C than when irradiated at 3 to 5 degrees C. Regardless of the temperature during irradiation, the level of fat did not have a significant effect on the D10 value. Salmonellae behaved like E. coli O157:H7 in low-fat beef, but temperature did not have a significant effect when the pathogen was irradiated in high-fat ground beef. Significantly higher D10 values were calculated for C. jejuni irradiated in frozen than in refrigerated low-fat beef. C. jejuni was more resistant to irradiation in low-fat beef than in high-fat beef when treatment was at -17 to -15 degrees C. Regardless of the fat level and temperature during inactivation, these pathogens were highly sensitive to gamma irradiation.(ABSTRACT TRUNCATED AT 250 WORDS)