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Escherichia coli strains expressing H12 antigens demonstrate an increased ability to attach to abiotic surfaces as compared with E. coli strains expressing H7 antigens

Goulter RM, Taran E, Gentle IR, Gobius KS, Dykes GA

Colloids Surf B Biointerfaces, 2014 Jul 1;119:90-8.
PMID: 24880987 DOI: 10.1016/j.colsurfb.2014.04.003

The role of Escherichia coli H antigens in hydrophobicity and attachment to glass, Teflon and stainless steel (SS) surfaces was investigated through construction of fliC knockout mutants in E. coli O157:H7, O1:H7 and O157:H12. Loss of FliC(H12) in E. coli O157:H12 decreased attachment to glass, Teflon and stainless steel surfaces (p<0.05). Complementing E. coli O157:H12 ΔfliC(H12) with cloned wildtype (wt) fliC(H12) restored attachment to wt levels. The loss of FliCH7 in E. coli O157:H7 and O1:H7 did not always alter attachment (p>0.05), but complementation with cloned fliC(H12), as opposed to cloned fliCH7, significantly increased attachment for both strains compared with wt counterparts (p<0.05). Hydrophobicity determined using bacterial adherence to hydrocarbons and contact angle measurements differed with fliC expression but was not correlated to the attachment to materials included in this study. Purified FliC was used to functionalise silicone nitride atomic force microscopy probes, which were used to measure adhesion forces between FliC and substrates. Although no significant difference in adhesion force was observed between FliC(H12) and FliCH7 probes, differences in force curves suggest different mechanism of attachment for FliC(H12) compared with FliCH7. These results indicate that E. coli strains expressing flagellar H12 antigens have an increased ability to attach to certain abiotic surfaces compared with E. coli strains expressing H7 antigens.
Fulltext Effect of frozen storage on the characteristics of a developed and commercial fish sausages

Al-Bulushi IM, Kasapis S, Dykes GA, Al-Waili H, Guizani N, Al-Oufi H

J Food Sci Technol, 2013 Dec;50(6):1158-64.
PMID: 24426029 DOI: 10.1007/s13197-011-0441-x

The effect of frozen storage on the physiochemical, chemical and microbial characteristics of two types of fish sausages was studied. Fish sausages developed (DFS) with a spice-sugar formulation and commercial fish sausages (CFS) were stored at -20 °C for 3 months. Fresh DFS contained 12.22% lipids and had a 3.53 cfu/g total bacteria count (TBC) whereas, CFS contained 5.5% lipids and had a 4.81 cfu/g TBC. During storage, TBC decreased significantly (p 0.05) in CFS. A peroxide value (PV) was not detectable until week four and eight of storage in CFS and DFS, respectively. The salt-soluble proteins (SSP) level was stable in DFS but in CFS it declined significantly (p 0.05) in both sausage types. This study showed that the effect of storage at -20 °C on fish sausages characteristics varied between formulations and depended on the ingredients of fish sausages.
Determination of quarantine period in African catfish (Clarias gariepinus) fed with pig (Sus sp.) offal to assure compliance with halal standards

Wan Norhana MN, Dykes GA, Padilah B, Ahmad Hazizi AA, Masazurah AR

Food Chem, 2012 Dec 1;135(3):1268-72.
PMID: 22953853 DOI: 10.1016/j.foodchem.2012.05.083

Pig (Sus sp.) and pig by-products are considered as najasa (impurities) in Islam and forbidden in Muslim consumer products. Animals fed on najasa are categorised as al-jallālah (contaminated animals) which are allowed to be consumed as long as they have been quarantined for a certain period of time. During this quarantine period the animals will have undergone a natural purification process or istihālah. African catfish (Clarias gariepinus) are commonly consumed in Malaysia and may be fed on najasa. This study was carried out to estimate the istihālah period for catfish after feeding with pig offal, based on the absence of pig DNA in catfish gut and to suggest the quarantine period in catfish fed with pig offal. The results indicated that the maximum istihālah period could reach 36h in the stomach, 6h in the midgut and less than 2h in the hindgut although in many cases shorter periods were observed. Based on these results it is estimated that the minimum quarantine period for catfish fed with pig offal is 1.5days.
Fulltext Genome sequencing and annotation of Cellulomonas sp. HZM

Chua P, Har ZM, Austin CM, Yule CM, Dykes GA, Lee SM

Genom Data, 2015 Sep;5:40-1.
PMID: 26484221 DOI: 10.1016/j.gdata.2015.05.009

We report the draft genome sequence of Cellulomonas sp. HZM, isolated from a tropical peat swamp forest. The draft genome size is 3,559,280 bp with a G + C content of 73% and contains 3 rRNA sequences (single copies of 5S, 16S and 23S rRNA).
Fulltext Genome sequencing and annotation of Aeromonas sp. HZM

Chua P, Har ZM, Austin CM, Yule CM, Dykes GA, Lee SM

Genom Data, 2015 Sep;5:38-9.
PMID: 26484220 DOI: 10.1016/j.gdata.2015.05.008

We report the draft genome sequence of Aeromonas sp. strain HZM, isolated from tropical peat swamp forest soil. The draft genome size is 4,451,364 bp with a G + C content of 61.7% and contains 10 rRNA sequences (eight copies of 5S rRNA genes, single copy of 16S and 23S rRNA each). The genome sequence can be accessed at DDBJ/EMBL/GenBank under the accession no. JEMQ00000000.
Fulltext Attachment of Salmonella strains to a plant cell wall model is modulated by surface characteristics and not by specific carbohydrate interactions

Tan MS, Moore SC, Tabor RF, Fegan N, Rahman S, Dykes GA

BMC Microbiol, 2016 09 15;16:212.
PMID: 27629769 DOI: 10.1186/s12866-016-0832-2

BACKGROUND: Processing of fresh produce exposes cut surfaces of plant cell walls that then become vulnerable to human foodborne pathogen attachment and contamination, particularly by Salmonella enterica. Plant cell walls are mainly composed of the polysaccharides cellulose, pectin and hemicelluloses (predominantly xyloglucan). Our previous work used bacterial cellulose-based plant cell wall models to study the interaction between Salmonella and the various plant cell wall components. We demonstrated that Salmonella attachment was favoured in the presence of pectin while xyloglucan had no effect on its attachment. Xyloglucan significantly increased the attachment of Salmonella cells to the plant cell wall model only when it was in association with pectin. In this study, we investigate whether the plant cell wall polysaccharides mediate Salmonella attachment to the bacterial cellulose-based plant cell wall models through specific carbohydrate interactions or through the effects of carbohydrates on the physical characteristics of the attachment surface.
RESULTS: We found that none of the monosaccharides that make up the plant cell wall polysaccharides specifically inhibit Salmonella attachment to the bacterial cellulose-based plant cell wall models. Confocal laser scanning microscopy showed that Salmonella cells can penetrate and attach within the tightly arranged bacterial cellulose network. Analysis of images obtained from atomic force microscopy revealed that the bacterial cellulose-pectin-xyloglucan composite with 0.3 % (w/v) xyloglucan, previously shown to have the highest number of Salmonella cells attached to it, had significantly thicker cellulose fibrils compared to other composites. Scanning electron microscopy images also showed that the bacterial cellulose and bacterial cellulose-xyloglucan composites were more porous when compared to the other composites containing pectin.
CONCLUSIONS: Our study found that the attachment of Salmonella cells to cut plant cell walls was not mediated by specific carbohydrate interactions. This suggests that the attachment of Salmonella strains to the plant cell wall models were more dependent on the structural characteristics of the attachment surface. Pectin reduces the porosity and space between cellulose fibrils, which then forms a matrix that is able to retain Salmonella cells within the bacterial cellulose network. When present with pectin, xyloglucan provides a greater surface for Salmonella cells to attach through the thickening of cellulose fibrils.
Fulltext Draft Genome Sequences of Six Enterococcus faecalis Strains Isolated from Malaysian Clinical and Environmental Origins

Daniel DS, Gan HM, Lee SM, Dykes GA, Rahman S

Genome Announc, 2017 Jun 15;5(24).
PMID: 28619812 DOI: 10.1128/genomeA.00553-17

Enterococcus faecalis is known to cause a variety of nosocomial infections, including urinary tract infections. Antibiotic resistance and virulence properties in this species are of public concern. The draft genome sequences of six E. faecalis strains isolated from clinical and environmental sources in Malaysia are presented here.
Fulltext Genetic diversity of Enterococcus faecalis isolated from environmental, animal and clinical sources in Malaysia

Daniel DS, Lee SM, Gan HM, Dykes GA, Rahman S

J Infect Public Health, 2017 02 21;10(5):617-623.
PMID: 28254461 DOI: 10.1016/j.jiph.2017.02.006

Enterococcus faecalis ranks as one of the leading causes of nosocomial infections. A strong epidemiological link has been reported between E. faecalis inhabiting animals and environmental sources. This study investigates the genetic diversity, antibiotic resistance and virulence determinants in E. faecalis from three sources in Malaysia. A total of 250 E. faecalis isolates were obtained consisting of 120 isolates from farm animals, 100 isolates from water sources and 30 isolates from hospitalized patients. Pulse-field gel electrophoresis-typing yielded 63 pulsotypes, with high diversity observed in all sources (D=≥0.901). No pulsotype was common to all the three sources. Each patient room had its own unique PFGE pattern which persisted after six months. Minimum inhibitory concentrations of Vancomycin, Gentamicin, Penicillin, Tetracycline, Nitrofurantoin, Levofloxacin, Ciprofloxacin and Fosfomycin were evaluated. Resistance to Tetracycline was most prevalent in isolates from farm animals (62%) and water sources (49%). Water isolates (86%) had a higher prevalence of the asa1 gene, which encodes for aggregation substance, whereas clinical (78%) and farm animal isolates (87%) had a higher prevalence of the esp gene, encoding a surface exposed protein. This study generates knowledge on the genetic diversity of E. faecalis with antibiotic resistance and virulence characteristics from various sources in Malaysia.
Fulltext Stochasticity of bacterial attachment and its predictability by the extended derjaguin-landau-verwey-overbeek theory

Chia TW, Nguyen VT, McMeekin T, Fegan N, Dykes GA

Appl Environ Microbiol, 2011 Jun;77(11):3757-64.
PMID: 21478319 DOI: 10.1128/AEM.01415-10

Bacterial attachment onto materials has been suggested to be stochastic by some authors but nonstochastic and based on surface properties by others. We investigated this by attaching pairwise combinations of two Salmonella enterica serovar Sofia (S. Sofia) strains (with different physicochemical and attachment properties) with one strain each of S. enterica serovar Typhimurium, S. enterica serovar Infantis, or S. enterica serovar Virchow (all with similar physicochemical and attachment abilities) in ratios of 0.428, 1, and 2.333 onto glass, stainless steel, Teflon, and polysulfone. Attached bacterial cells were recovered and counted. If the ratio of attached cells of each Salmonella serovar pair recovered was the same as the initial inoculum ratio, the attachment process was deemed stochastic. Experimental outcomes from the study were compared to those predicted by the extended Derjaguin-Landau-Verwey-Overbeek (XDLVO) theory. Significant differences (P < 0.05) between the initial and the attached ratios for serovar pairs containing S. Sofia S1296a for all different ratios were apparent for all materials. For S. Sofia S1635-containing pairs, 7 out of 12 combinations of serovar pairs and materials had attachment ratios not significantly different (P > 0.05) from the initial ratio of 0.428. Five out of 12 and 10 out of 12 samples had attachment ratios not significantly different (P > 0.05) from the initial ratios of 1 and 2.333, respectively. These results demonstrate that bacterial attachment to different materials is likely to be nonstochastic only when the key physicochemical properties of the bacteria were significantly different (P < 0.05) from each other. XDLVO theory could successfully predict the attachment of some individual isolates to particular materials but could not be used to predict the likelihood of stochasticity in pairwise attachment experiments.
Fulltext Inhibition of attachment of oral bacteria to immortalized human gingival fibroblasts (HGF-1) by tea extracts and tea components

Wang Y, Chung FF, Lee SM, Dykes GA

BMC Res Notes, 2013;6:143.
PMID: 23578062 DOI: 10.1186/1756-0500-6-143

Tea has been suggested to promote oral health by inhibiting bacterial attachment to the oral cavity. Most studies have focused on prevention of bacterial attachment to hard surfaces such as enamel.
Enhanced xylose recovery from oil palm empty fruit bunch by efficient acid hydrolysis

Tan HT, Dykes GA, Wu TY, Siow LF

Appl Biochem Biotechnol, 2013 Aug;170(7):1602-13.
PMID: 23709290 DOI: 10.1007/s12010-013-0298-8

Oil palm empty fruit bunch (EFB) is abundantly available in Malaysia and it is a potential source of xylose for the production of high-value added products. This study aimed to optimize the hydrolysis of EFB using dilute sulfuric acid (H2SO4) and phosphoric acid (H3PO4) via response surface methodology for maximum xylose recovery. Hydrolysis was carried out in an autoclave. An optimum xylose yield of 91.2 % was obtained at 116 °C using 2.0 % (v/v) H2SO4, a solid/liquid ratio of 1:5 and a hydrolysis time of 20 min. A lower optimum xylose yield of 24.0 % was observed for dilute H3PO4 hydrolysis at 116 °C using 2.4 % (v/v) H3PO4, a solid/liquid ratio of 1:5 and a hydrolysis time of 20 min. The optimized hydrolysis conditions suggested that EFB hydrolysis by H2SO4 resulted in a higher xylose yield at a lower acid concentration as compared to H3PO4.
Nepenthes ampullaria (Nepenthaceae) Pitchers Are Unattractive to Gravid Aedes aegypti and Aedes albopictus (Diptera: Culicidae)

Chou LY, Dykes GA, Wilson RF, Clarke CM

Environ Entomol, 2016 Feb;45(1):201-6.
PMID: 26518035 DOI: 10.1093/ee/nvv164

Nepenthes pitcher plants are colonized by a variety of specialized arthropods. As Aedes mosquitoes are container breeders, Nepenthes pitchers are a potential candidate oviposition site for vector species, such as Aedes aegypti (L.) and Aedes albopictus (Skuse). However, Aedes spp. are not commonly encountered in Nepenthes pitchers, and the environment inside the pitchers of some species is lethal to them. One exception is Nepenthes ampullaria Jack, whose pitchers are known to be colonized by Ae. albopictus on very rare occasions. Given that Ae. albopictus larvae can survive in N. ampullaria pitcher fluids, we sought to determine why pitcher colonization is rare, testing the hypothesis that gravid Aedes mosquitoes are deterred from ovipositing into container habitats that have similar characteristics to N. ampullaria pitchers. Using plastic ovitraps of different sizes, colors, and with different types of fluids (based on the characteristics of N. ampullaria pitchers), we compared oviposition rates by Aedes mosquitoes in urban and rural areas within the geographical range of N. ampullaria near Kuala Lumpur, Malaysia. Ovitraps that were black and large (>250-ml capacity) accumulated significantly more eggs than ovitraps that were smaller, or green in color. In terms of size and color, small, green ovitraps are analogous to N. ampullaria pitchers, indicating that these pitchers are not particularly attractive to gravid Ae. albopictus. Although Aedes spp. are capable of colonizing N. ampullaria pitchers, the pitchers are relatively unattractive to gravid females and do not represent a significant habitat for larvae of dengue vectors at present.
Fulltext Characterization of Shiga Toxigenic Escherichia coli O157 and Non-O157 Isolates from Ruminant Feces in Malaysia

Perera A, Clarke CM, Dykes GA, Fegan N

Biomed Res Int, 2015;2015:382403.
PMID: 26539484 DOI: 10.1155/2015/382403

Shiga toxigenic Escherichia coli (STEC) O157 and several other serogroups of non-O157 STEC are causative agents of severe disease in humans world-wide. The present study was conducted to characterize STEC O157 and non-O157 serogroups O26, O103, O111, O121, O45, and O145 in ruminants in Malaysia. A total of 136 ruminant feces samples were collected from 6 different farms in Peninsular Malaysia. Immunomagnetic beads were used to isolate E. coli O157 and non-O157 serogroups, while PCR was used for the detection and subtyping of STEC isolates. STEC O157:H7 was isolated from 6 (4%) feces samples and all isolates obtained carried stx 2c, eaeA-γ1, and ehxA. Non-O157 STEC was isolated from 2 (1.5%) feces samples with one isolate carrying stx 1a, stx 2a, stx 2c, and ehxA and the other carrying stx 1a alone. The presence of STEC O157 and non-O157 in a small percentage of ruminants in this study together with their virulence characteristics suggests that they may have limited impact on public health.
Fulltext Draft Genome Sequences of Three Multiantibiotic-Resistant Campylobacter jejuni Strains (2865, 2868, and 2871) Isolated from Poultry at Retail Outlets in Malaysia

Teh AH, Lee SM, Dykes GA

Genome Announc, 2016;4(3).
PMID: 27151799 DOI: 10.1128/genomeA.00331-16

Campylobacter jejuni is a frequent cause of human bacterial gastrointestinal foodborne disease worldwide. Antibiotic resistance in this species is of public health concern. The draft genome sequences of three multiantibiotic-resistant C. jejuni strains (2865, 2868, and 2871) isolated from poultry at retail outlets in Malaysia are presented here.
Growth in the presence of specific antibiotics induces biofilm formation by a Campylobacter jejuni strain sensitive to them but not in resistant strains

Teh AHT, Lee SM, Dykes GA

J Glob Antimicrob Resist, 2019 09;18:55-58.
PMID: 31163253 DOI: 10.1016/j.jgar.2019.05.020

OBJECTIVE: Campylobacter jejuni (C. jejuni) are among the most frequently identified bacteria associated with human gastroenteritis worldwide. Exposure to antibiotics may induce or inhibit biofilm formation in some bacterial species. Little work has been reported on the influence of antibiotics on biofilm formation by C. jejuni.
METHODS: This study investigated the effect of six different classes of antibiotics with different modes of action (ampicillin, ciprofloxacin, erythromycin, nalidixic acid, rifampicin and tetracycline) on biofilm formation in vitro by seven C. jejuni from poultry with different antibiotic resistance profiles.
RESULTS: The results indicated that in the presence of most of the tested antibiotics, biofilm formation by C. jejuni strains, which are resistant to them, was reduced but biofilm formation in sensitive strains was increased.
CONCLUSION: The ability of certain antibiotics to induce biofilm formation by a tested C. jejuni strain is of concern, with respect to the effective control of disease caused by this pathogen; however, further work is required to confirm how widespread this feature is.
A statistical approach for modelling the physical process of bacterial attachment to abiotic surfaces

Wang Y, Lee SM, Gentle IR, Dykes GA

Biofouling, 2020 11;36(10):1227-1242.
PMID: 33412938 DOI: 10.1080/08927014.2020.1865934

A statistical approach using a polynomial linear model in combination with a probability distribution model was developed to mathematically represent the process of bacterial attachment and study its mechanism. The linear deterministic model was built based on data from experiments investigating bacterial and substratum surface physico-chemical factors as predictors of attachment. The prediction results were applied to a normal-approximated binomial distribution model to probabilistically predict attachment. The experimental protocol used mixtures of Streptococcus salivarius and Escherichia coli, and mixtures of porous poly(butyl methacrylate-co-ethyl dimethacrylate) and aluminum sec-butoxide coatings, at varying ratios, to allow bacterial attachment to substratum surfaces across a range of physico-chemical properties (including the surface hydrophobicity of bacterial cells and the substratum, the surface charge of the cells and the substratum, the substratum surface roughness and cell size). The model was tested using data from independent experiments. The model indicated that hydrophobic interaction was the most important predictor while reciprocal interactions existed between some of the factors. More importantly, the model established a range for each factor within which the resultant attachment is unpredictable. This model, however, considers bacterial cells as colloidal particles and accounts only for the essential physico-chemical attributes of the bacterial cells and substratum surfaces. It is therefore limited by a lack of consideration of biological and environmental factors. This makes the model applicable only to specific environments and potentially provides a direction to future modelling for different environments.
The Influence of Prior Modes of Growth, Temperature, Medium, and Substrate Surface on Biofilm Formation by Antibiotic-Resistant Campylobacter jejuni

Teh AH, Lee SM, Dykes GA

Curr Microbiol, 2016 Dec;73(6):859-866.
PMID: 27623781

Campylobacter jejuni is one of the most common causes of bacterial gastrointestinal food-borne infection worldwide. It has been suggested that biofilm formation may play a role in survival of these bacteria in the environment. In this study, the influence of prior modes of growth (planktonic or sessile), temperatures (37 and 42 °C), and nutrient conditions (nutrient broth and Mueller-Hinton broth) on biofilm formation by eight C. jejuni strains with different antibiotic resistance profiles was examined. The ability of these strains to form biofilm on different abiotic surfaces (stainless steel, glass, and polystyrene) as well as factors potentially associated with biofilm formation (bacterial surface hydrophobicity, auto-aggregation, and initial attachment) was also determined. The results showed that cells grown as sessile culture generally have a greater ability to form biofilm (P
Fulltext Association of some Campylobacter jejuni with Pseudomonas aeruginosa biofilms increases attachment under conditions mimicking those in the environment

Teh AHT, Lee SM, Dykes GA

PLoS One, 2019;14(4):e0215275.
PMID: 30970009 DOI: 10.1371/journal.pone.0215275

Campylobacter jejuni is a microaerophilic bacterial species which is a major food-borne pathogen worldwide. Attachment and biofilm formation have been suggested to contribute to the survival of this fastidious bacteria in the environment. In this study the attachment of three C. jejuni strains (C. jejuni strains 2868 and 2871 isolated from poultry and ATCC 33291) to different abiotic surfaces (stainless steel, glass and polystyrene) alone or with Pseudomonas aeruginosa biofilms on them, in air at 25°C and under static or flow conditions, were investigated using a modified Robbins Device. Bacteria were enumerated and scanning electron microscopy was carried out. The results indicated that both C. jejuni strains isolated from poultry attached better to Pseudomonas aeruginosa biofilms on abiotic surfaces than to the surfaces alone under the different conditions tested. This suggests that biofilms of other bacterial species may passively protect C. jejuni against shear forces and potentially oxygen stress which then contribute to their persistence in environments which are detrimental to them. By contrast the C. jejuni ATCC 33291 strain did not attach differentially to P. aeruginosa biofilms, suggesting that different C. jejuni strains may have alternative strategies for persistence in the environment. This study supports the hypothesis that C. jejuni do not form biofilms per se under conditions they encounter in the environment but simply attach to surfaces or biofilms of other species.
The influence of dissolved oxygen level and medium on biofilm formation by Campylobacter jejuni

Teh AHT, Lee SM, Dykes GA

Food Microbiol, 2017 Feb;61:120-125.
PMID: 27697161 DOI: 10.1016/j.fm.2016.09.008

Campylobacter jejuni survival in aerobic environments has been suggested to be mediated by biofilm formation. Biofilm formation by eight C. jejuni strains under both aerobic and microaerobic conditions in different broths (Mueller-Hinton (MH), Bolton and Brucella) was quantified. The dissolved oxygen (DO) content of the broths under both incubation atmospheres was determined. Biofilm formation for all strains was highest in MH broth under both incubation atmospheres. Four strains had lower biofilm formation in MH under aerobic as compared to microaerobic incubation, while biofilm formation by the other four strains did not differ under the 2 atm. Two strains had higher biofilm formation under aerobic as compared to microaerobic atmospheres in Bolton broth. Biofilm formation by all other strains in Bolton, and all strains in Brucella broth, did not differ under the 2 atm. Under aerobic incubation DO levels in MH > Brucella > Bolton broth. Under microaerobic conditions levels in MH = Brucella > Bolton broth. Levels of DO in MH and Brucella broth were lower under microaerobic conditions but those of Bolton did not differ under the 2 atm. Experimental conditions and especially the DO of broth media confound previous conclusions drawn about aerobic biofilm formation by C. jejuni.
Fulltext Identification of potential Campylobacter jejuni genes involved in biofilm formation by EZ-Tn5 Transposome mutagenesis

Teh AHT, Lee SM, Dykes GA

BMC Res Notes, 2017 May 12;10(1):182.
PMID: 28499399 DOI: 10.1186/s13104-017-2504-1

BACKGROUND: Biofilm formation has been suggested to play a role in the survival of Campylobacter jejuni in the environment and contribute to the high incidence of human campylobacteriosis. Molecular studies of biofilm formation by Campylobacter are sparse.
RESULTS: We attempted to identify genes that may be involved in biofilm formation in seven C. jejuni strains through construction of mutants using the EZ-Tn5 Transposome system. Only 14 mutants with reduced biofilm formation were obtained, all from one strain of C. jejuni. Three different genes of interest, namely CmeB (synthesis of multidrug efflux system transporter proteins), NusG (transcription termination and anti-termination protein) and a putative transmembrane protein (involved in membrane protein function) were identified. The efficiency of the EZ::TN5 transposon mutagenesis approach was strain dependent and was unable to generate any mutants from most of the strains used.
CONCLUSIONS: A diverse range of genes may be involved in biofilm formation by C. jejuni. The application of the EZ::TN5 system for construction of mutants in different Campylobacter strains is limited.

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