Diluents and the Enumeration of Stressed Campylobacter jejuni

Significant discrepancies in calculated counts of Campylobacter jejuni strains ATCC 33250 and 29428 were observed between undiluted samples and samples diluted in 0.1% peptone water when cultures had been incubated at 42°C under micro-aerobic conditions in brucella broth containing 2% NaCl. The influence of 0.1% peptone water, brucella broth and phosphate buffer as diluents for enumeration of C. jejuni stored at 6 and −18°C in brucella broth with 0.5 and 2% NaCl was studied. The three diluents were equally effective for the recovery of C. jejuni from refrigerated broth containing 0.5% NaCl. However, when the organism had been refrigerated in the broth with 2% NaCl or frozen in the broth with either level of salt, dilution with brucella broth produced significantly higher counts than dilution with 0.1% peptone water or phosphate buffer.

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Biofilm Formation by Campylobacter jejuni Is Increased under Aerobic Conditions

Applied and Environmental Microbiology ◽

10.1128/aem.01878-09 ◽

2010 ◽

Vol 76 (7) ◽

pp. 2122-2128 ◽

Cited By ~ 122

Author(s):

Mark Reuter ◽

Arthur Mallett ◽

Bruce M. Pearson ◽

Arnoud H. M. van Vliet

Keyword(s):

Campylobacter Jejuni ◽

Food Chain ◽

Biofilm Formation ◽

Planktonic Cells ◽

Aerobic Conditions ◽

Food Borne ◽

Passive Release ◽

Bacterial Gastroenteritis ◽

Microaerobic Conditions ◽

Developed World

ABSTRACT The microaerophilic human pathogen Campylobacter jejuni is the leading cause of food-borne bacterial gastroenteritis in the developed world. During transmission through the food chain and the environment, the organism must survive stressful environmental conditions, particularly high oxygen levels. Biofilm formation has been suggested to play a role in the environmental survival of this organism. In this work we show that C. jejuni NCTC 11168 biofilms developed more rapidly under environmental and food-chain-relevant aerobic conditions (20% O2) than under microaerobic conditions (5% O2, 10% CO2), although final levels of biofilms were comparable after 3 days. Staining of biofilms with Congo red gave results similar to those obtained with the commonly used crystal violet staining. The level of biofilm formation by nonmotile aflagellate strains was lower than that observed for the motile flagellated strain but nonetheless increased under aerobic conditions, suggesting the presence of flagellum-dependent and flagellum-independent mechanisms of biofilm formation in C. jejuni. Moreover, preformed biofilms shed high numbers of viable C. jejuni cells into the culture supernatant independently of the oxygen concentration, suggesting a continuous passive release of cells into the medium rather than a condition-specific active mechanism of dispersal. We conclude that under aerobic or stressful conditions, C. jejuni adapts to a biofilm lifestyle, allowing survival under detrimental conditions, and that such a biofilm can function as a reservoir of viable planktonic cells. The increased level of biofilm formation under aerobic conditions is likely to be an adaptation contributing to the zoonotic lifestyle of C. jejuni.

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Evaluation of an Enrichment-Plating Procedure for the Recovery of Campylobacter jejuni from Turkey Eggs and Meat

Journal of Food Protection ◽

10.4315/0362-028x-45.14.1276 ◽

1982 ◽

Vol 45 (14) ◽

pp. 1276-1278 ◽

Cited By ~ 12

Author(s):

G. R. ACUFF ◽

C. VANDERZANT ◽

F. A. GARDNER ◽

F. A. GOLAN

Keyword(s):

Campylobacter Jejuni ◽

Ferrous Sulfate ◽

Antimicrobial Agents ◽

Sodium Metabisulfite ◽

Sodium Pyruvate ◽

Enrichment Broth ◽

Initial Cell ◽

Selective Enrichment ◽

Cell Numbers ◽

Brucella Broth

A selective enrichment-plating procedure was tested for the recovery and enumeration of Campylobacter jejuni from turkey eggs and meat. Enrichment was in brucella broth with ferrous sulfate, sodium metabisulfite, sodium pyruvate and five antimicrobial agents. Plating was on brucella agar supplemented with equine blood and antimicrobial agents. Incubation of tubes and plates was at 42°C in an atmosphere of 5% O2:10% CO2:85% N2. C. jejuni could be recovered from the enrichment broth when calculated initial cell numbers per ml of broth were as low as 0.3 to 3.3

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Chicken Juice Enhances Surface Attachment and Biofilm Formation of Campylobacter jejuni

Applied and Environmental Microbiology ◽

10.1128/aem.02614-14 ◽

2014 ◽

Vol 80 (22) ◽

pp. 7053-7060 ◽

Cited By ~ 68

Author(s):

Helen L. Brown ◽

Mark Reuter ◽

Louise J. Salt ◽

Kathryn L. Cross ◽

Roy P. Betts ◽

...

Keyword(s):

Campylobacter Jejuni ◽

Food Chain ◽

Biofilm Formation ◽

Cell Attachment ◽

Bacterial Attachment ◽

Poultry Meat ◽

Abiotic Surface ◽

Surface Attachment ◽

Aerobic Conditions ◽

Content Type

ABSTRACTThe bacterial pathogenCampylobacter jejuniis primarily transmitted via the consumption of contaminated foodstuffs, especially poultry meat. In food processing environments,C. jejuniis required to survive a multitude of stresses and requires the use of specific survival mechanisms, such as biofilms. An initial step in biofilm formation is bacterial attachment to a surface. Here, we investigated the effects of a chicken meat exudate (chicken juice) onC. jejunisurface attachment and biofilm formation. Supplementation of brucella broth with ≥5% chicken juice resulted in increased biofilm formation on glass, polystyrene, and stainless steel surfaces with fourC. jejuniisolates and oneC. coliisolate in both microaerobic and aerobic conditions. When incubated with chicken juice,C. jejuniwas both able to grow and form biofilms in static cultures in aerobic conditions. Electron microscopy showed thatC. jejunicells were associated with chicken juice particulates attached to the abiotic surface rather than the surface itself. This suggests that chicken juice contributes toC. jejunibiofilm formation by covering and conditioning the abiotic surface and is a source of nutrients. Chicken juice was able to complement the reduction in biofilm formation of an aflagellated mutant ofC. jejuni, indicating that chicken juice may support food chain transmission of isolates with lowered motility. We provide here a useful model for studying the interaction ofC. jejunibiofilms in food chain-relevant conditions and also show a possible mechanism forC. jejunicell attachment and biofilm initiation on abiotic surfaces within the food chain.

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Kinetic Behavior of Campylobacter jejuni in Beef Tartare at Cold Temperatures and Transcriptomes Related to Its Survival

Journal of Food Protection ◽

10.4315/0362-028x.jfp-17-236 ◽

2017 ◽

Vol 80 (12) ◽

pp. 2127-2131 ◽

Cited By ~ 2

Author(s):

Sejeong Kim ◽

Jiyeon Jeong ◽

Heeyoung Lee ◽

Jeeyeon Lee ◽

Soomin Lee ◽

...

Keyword(s):

Oxidative Stress ◽

Stress Response ◽

Campylobacter Jejuni ◽

Survival Data ◽

Weibull Model ◽

Effect Of Temperature ◽

Kinetic Behavior ◽

Cold Temperatures ◽

Aerobic Conditions ◽

Stress Response Genes

ABSTRACT This study was conducted to examine the kinetic behavior of Campylobacter jejuni in raw beef tartare by using mathematical models and to identify genes related to C. jejuni survival at cold temperatures. C. jejuni was inoculated onto beef tartare samples, stored at 4, 10, 15, 25, and 30°C, plated on modified charcoal-cefoperazone-deoxycholate agar, and enumerated. The survival data was fitted to the Weibull model to calculate delta (δ), which is the time required for the first 1-log reduction of the cells. The Davey model was then fitted to the δ to evaluate the effect of temperature. To evaluate the performance of the developed model, the root mean square error (RMSE) was calculated by comparing the observed data with the predicted data. The mRNA was extracted from samples stored at 4 and 30°C under aerobic and anaerobic conditions, and the expression of oxidative stress response and stress response genes was evaluated. C. jejuni survived in beef tartare longer at 4°C (δ = 657.1 ± 79.6 min) than at other temperatures (9.7 ± 11.2 to 465.7 ± 139.3°C) even under aerobic conditions. The RMSE (0.475) suggested that the developed model was appropriate to describe the kinetic behavior of C. jejuni. Quantitative real-time PCR results revealed that oxidative stress and stress response genes were related to C. jejuni survival at cold temperatures, even under aerobic conditions. These results indicate that the model will be useful for describing the kinetic behavior of C. jejuni in beef tartare and that this pathogen can survive at cold temperatures because of the expression of the sodB, katA, and clpP genes.

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Campylobacter jejuni Biofilm Formation Under Aerobic Conditions and Inhibition by ZnO Nanoparticles

Frontiers in Microbiology ◽

10.3389/fmicb.2020.00207 ◽

2020 ◽

Vol 11 ◽

Cited By ~ 3

Author(s):

Xian Zhong ◽

Qingping Wu ◽

Jumei Zhang ◽

Zonghao Ma ◽

Juan Wang ◽

...

Keyword(s):

Campylobacter Jejuni ◽

Zno Nanoparticles ◽

Biofilm Formation ◽

Aerobic Conditions

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Adhesion, Biofilm Formation, and Genomic Features of Campylobacter jejuni Bf, an Atypical Strain Able to Grow under Aerobic Conditions

Frontiers in Microbiology ◽

10.3389/fmicb.2016.01002 ◽

2016 ◽

Vol 7 ◽

Cited By ~ 19

Author(s):

Vicky Bronnec ◽

Hana Turoňová ◽

Agnès Bouju ◽

Stéphane Cruveiller ◽

Ramila Rodrigues ◽

...

Keyword(s):

Campylobacter Jejuni ◽

Biofilm Formation ◽

Aerobic Conditions ◽

Genomic Features

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Use of Luminescent Campylobacter jejuni ATCC 33291 To Assess Eggshell Colonization and Penetration in Fresh and Retail Eggs

Journal of Food Protection ◽

10.4315/0362-028x-64.12.2058 ◽

2001 ◽

Vol 64 (12) ◽

pp. 2058-2062 ◽

Cited By ~ 25

Author(s):

KEVIN J. ALLEN ◽

MANSEL W. GRIFFITHS

Keyword(s):

Electron Microscopy ◽

Scanning Electron Microscopy ◽

Campylobacter Jejuni ◽

Effective Means ◽

Photon Counting ◽

Transcriptional Fusion ◽

Charge Coupled Device ◽

Aerobic Conditions ◽

Microaerophilic Conditions ◽

Scanning Electron

A luminescent phenotype in Campylobacter jejuni ATCC 33291, generated by a transcriptional fusion between the C. jejuni flaA σ28 promoter and the luxCDABE genes of Xenorhabdus luminescens on plasmid pRYluxCDABE, was used to examine colonization and penetration of fresh and retail eggs. C. jejuni colonized both fresh and retail eggs at 37, 40, and 42°C under microaerophilic conditions. Fresh eggs were more heavily colonized than retail eggs. Under aerobic conditions, fresh eggs were colonized at similar levels for all three temperatures. C. jejuni was found to penetrate the eggshell in 2 of 48 (4.2%) fresh eggs assessed. Although the lux+ phenotype did not provide an effective means of predicting penetration sites, it was effective at visualizing eggshell colonization. Also, it effectively demonstrated the organism's opportunistic nature, as eggshell surfaces with flaws and slight cracks were extensively colonized and easily detected by a photon counting charge-coupled device camera. Using scanning electron microscopy, C. jejuni ATCC 33291 was visualized on both the exterior and interior surfaces of the egg membranes indicating penetration of these barriers.

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Effectiveness of Chemical Sanitizers against Campylobacter jejuni–Containing Biofilms

Journal of Food Protection ◽

10.4315/0362-028x-65.7.1117 ◽

2002 ◽

Vol 65 (7) ◽

pp. 1117-1121 ◽

Cited By ~ 63

Author(s):

NATHANON TRACHOO ◽

JOSEPH F. FRANK

Keyword(s):

Mixed Culture ◽

Polyvinyl Chloride ◽

Campylobacter Jejuni ◽

Peracetic Acid ◽

Glass Beads ◽

Acid Mixture ◽

Pseudomonas Sp ◽

Gram Positive ◽

Brucella Broth ◽

Chicken House

Survival of Campylobacter jejuni in mixed-culture biofilms was determined after treatment with chemical sanitizers including chlorine, quaternary ammonia, peracetic acid (PAA), and a PAA/peroctanoic acid mixture (PAA/POA). Biofilm-producing bacteria (gram-positive rods, Y1 and W1) were isolated from chicken house nipple drinkers. A meat plant isolate (Pseudomonas sp.) was also included as a biofilm producer. Two-day-old biofilms grown on polyvinyl chloride (PVC) plastic coupons in R2A broth at 12°C were incubated with 106 CFU/ml C. jejuni for 6 h to allow attachment. The coupons were then rinsed and incubated in fresh media for an additional 24 h. C. jejuni–containing biofilms were detached by vortexing with glass beads in modified brucella broth, which was then enumerated for C. jejuni on selective/differential media. The presence of biofilm enhanced (P < 0.01) the attachment and survival of C. jejuni. After the 24-h incubation, only 20 CFU/cm2 of C. jejuni were recovered from the control without biofilms compared to 2,500 to 5,000 CFU/cm2 in samples with preexisting biofilms. The presence of biofilm microflora decreased (P < 0.01) the effectiveness of sanitizers against C. jejuni. Chlorine was the most effective sanitizer since it completely inactivated C. jejuni in the biofilms after treatment at 50 ppm for 45 s. C. jejuni in biofilms was susceptible to all sanitizers tested but was not completely inactivated by treatment with quaternary ammonia, PAA, or PAA/POA mixture at 50 and 200 ppm for 45 s.

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Association of glycerylphosphorylcholine with human sperm and effect of capacitation on their metabolism

Reproduction Fertility and Development ◽

10.1071/rd9940679 ◽

1994 ◽

Vol 6 (6) ◽

pp. 679 ◽

Cited By ~ 3

Author(s):

J Mitra ◽

M Chowdhury

Keyword(s):

Oxidative Phosphorylation ◽

Phosphate Buffer ◽

Sodium Phosphate ◽

Human Sperm ◽

Cervical Mucus ◽

Breakdown Product ◽

O2 Uptake ◽

Head Region ◽

Aerobic Conditions ◽

Sodium Phosphate Buffer

The presence and localization of glycerylphosphorylcholine (GPC) on the surface of human sperm, as well as the metabolism of its breakdown product L-glycerol 3 phosphate (G3P), were investigated. GPC was found to be associated with sperm after penetrating cervical mucus and was present after repeated washing of the sperm. GPC was partially released by treatment with 0.4 M NaCl in 0.01 M sodium phosphate buffer (pH 7.4) and localized to the head region after sperm fractionation. G3P did not increase O2 uptake of uncapacitated human sperm. However, under aerobic conditions, lactate accumulated when exogenous G3P or uterine GPC diesterase was added to sperm in suspension. The uptake of O2 by washed capacitated sperm pre-incubated with 1 unit of rat uterine GPC diesterase for 30 min was significant. This effect was inhibited by 2 microM oligomycin indicating that oxidative phosphorylation had occurred. The present study indicates that GPC may play a role in the metabolism of human sperm after capacitation.

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