Modulation of cell surface hydrophobicity and attachment of bacteria to abiotic surfaces and shrimp by malaysian herb extracts

The use of simple crude water extracts of common herbs to reduce bacterial attachment may be a cost-effective way to control bacterial foodborne pathogens, particularly in developing countries. The ability of water extracts of three common Malaysian herbs (Andrographis paniculata, Eurycoma longifolia, and Garcinia atroviridis) to modulate hydrophobicity and attachment to surfaces of five food-related bacterial strains (Bacillus cereus ATCC 14576, Escherichia coli ATCC 25922, Pseudomonas aeruginosa ATCC 10145, Salmonella Enteritidis ATCC 13076, Staphylococcus aureus ATCC 25923) were determined. The bacterial attachment to hydrocarbon assay was used to determine bacterial hydrophobicity. Staining and direct microscopic counts were used to determine attachment of bacteria to glass and stainless steel. Plating on selective media was used to determine attachment of bacteria to shrimp. All extracts were capable of either significantly ( P < 0.05) increasing or decreasing bacterial surface hydrophobicity, depending on the herb extract and bacteria combination. Bacterial attachment to all surfaces was either significantly (P < 0.05) increased or decreased, depending on the herb extract and bacteria combination. Overall, hydrophobicity did not show a significant correlation (P > 0.05) to bacterial attachment. For specific combinations of bacteria, surface material, and plant extract, significant correlations (R > 0.80) between hydrophobicity and attachment were observed. The highest of these was observed for S. aureus attachment to stainless steel and glass after treatment with the E. longifolia extract (R = 0.99, P < 0.01). The crude water herb extracts in this study were shown to have the potential to modulate specific bacterial and surface interactions and may, with further work, be useful for the simple and practical control of foodborne pathogens.     

Campylobacter contamination of broiler caeca and carcasses at the slaughterhouse and correlation with Salmonella contamination

In order to estimate the prevalence of Campylobacter spp. and Salmonella spp. on broiler chicken carcasses and the prevalence of Campylobacter spp. in caeca, 58 French slaughterhouses were investigated in 2008. Enumeration of Campylobacter spp. was also performed in order to study the relation between caeca and carcass contamination. A pool of 10 caeca and one carcass were collected from 425 different batches over a 12-month period in 2008. Salmonella was isolated on 32 carcasses leading to a prevalence of 7.5% ([5.0-10.0]_95%CI). The prevalence of Campylobacter was 77.2% ([73.2-81.2]_95%CI) in caeca and 87.5% ([84.4-90.7]_95%CI) on carcasses. No significant correlation was found between Campylobacter and Salmonella. Positive values of Campylobacter were normally distributed and the average level was 8.05 log₁₀ cfu/g ([7.94-8.16]_95%CI) in caeca and 2.39 cfu/g ([2.30-2.48]_95%CI) on carcasses. A positive correlation (r = 0.59) was found between the mean of Campylobacter in caeca and on carcasses (p < 0.001). Thus, carcasses from batches with Campylobacter-positive caeca had significantly (p < 0.001) higher numbers of Campylobacter per gram than batches with negative caeca. These results show that Campylobacter can be present in both matrices and reduction in caeca could be a possible way to reduce the amount of bacteria on carcasses. Of the 2504 identifications performed, 3 species of Campylobacter (Campylobacter jejuni, Campylobacter coli and Campylobacter lari) were identified. The main species recovered were C. jejuni and C. coli, which were isolated in 55.3% and 44.5% of positive samples, respectively. These two species were equally represented in caeca but C. jejuni was the most frequently isolated on carcasses with 57.1% and 42.5% of positive carcasses for C. jejuni and C. coli, respectively. This study underlines that target a reduction of Campylobacter on final products requires a decrease of contamination in caeca.     

Seasonal influence on the prevalence of thermotolerant Campylobacter in retail broiler meat in Denmark

In Denmark, the incidence of human campylobacteriosis cases, as well as the Campylobacter prevalence in broiler flocks, is strongly influenced by season with a summer peak in July-August. Therefore, it was considered that the prevalence of Campylobacter in broiler meat sold at retail in Denmark might also be influenced by season. A retrospective survey analysis was performed on 2001-2007 national surveillance data of the prevalence of thermotolerant Campylobacter in all conventional broiler flocks at slaughter, and in randomly sampled broiler meat at retail. There was a significant effect of season on the occurrence of Campylobacter in meat at retail; the largest effect was found for domestic chilled meat. Thus, the Campylobacter prevalence in Danish broiler flocks, which fluctuated with season, was found to be a strong predictor for the occurrence of Campylobacter in fresh, chilled, Danish broiler meat. However, besides flock prevalence, there was also a direct effect of season on the occurrence of Campylobacter in Danish broiler meat at retail.     

Resistances to benzalkonium chloride of bacteria dried with food elements on stainless steel surface

To confirm the importance of washing food sediments from the surface of food-related environments, we examined resistances against benzalkonium chloride of pathogenic bacterial (Escherichia coli O26, Pseudomonas aeruginosa and Staphylococcus aureus) cells dried and adhered on stainless steel dishes with milk, beef gravy or tuna gravy. Suspensions (0.1 ml) of these bacteria (8-9 log cfu/ml) were put on a 5 cm ? stainless steel dish and dried at room temperature (20-24 °C) for 90 min in a bio-clean bench with ventilation. Though these bacteria suspended with distilled water decreased 30-40 fold during the drying period, milk and the gravies protected the bacteria. Without the food elements, the adhered E. coli and Stap. aureus were decreased from 6 to<2 log cfu/dish by 0.5 mg/ml benzalkonium chloride (BKC) for 10 min treatment. Although Ps. aeruginosa showed resistance to BKC, the adhered cells were inactivated by 2.0 mg/ml BKC. However, the bactericidal effect disappeared by the food elements, particularly with milk, even at 1.0 and/or 2.0 mg/ml BKC levels. The protective efficiency of milk on bacteria disappeared if washed with water.     

Factors associated with carcass contamination by Campylobacter at slaughterhouse in cecal-carrier broilers

A study was conducted in 2009 to identify risk factors of Campylobacter spp. transmission from the digestive tract to the carcasses of standard broilers (slaughter age: 37 day, carcass weight: 1.3 kg on average). Counts of Campylobacter were performed on pools of 10 ceca and 10 neck-skins from 108 Campylobacter ceca-positive batches in three slaughterhouses. Technical and health data also was collected on the broilers: age, size, carcass weight (mean and standard deviation), condemnation rate, mortality rate and nature of treatment during the rearing period.Cecal counts varied from 4.8 to 10.2 log₁₀ cfu/g. In seventeen batches (15.7%), the skin count was below the detection limit. In the 91 batches with positive neck-skin test results, the counts varied from 2.0 to 5.2 log₁₀ cfu/g. Standard deviation of carcass weight, condemnation rate, slaughter rate and cecal count were significantly lower and growth rate higher in the 17 batches where neck-skin results were not detected positive. Multivariate analysis showed that batches with higher standard deviation of carcass weight were 5 to 9 fold more at risk of having detectable carcass contamination. Among the 91 positive neck-skin batches, only slaughter rate and cecal counts were found to have a significant but limited effect on the level of neck-skin contamination. As far as body weight homogeneity may be affected by disease, better health control can contribute to a reduction of the contamination of the broiler carcasses in Campylobacter carrier batches.     

Effect of temperature and growth media on the attachment of Listeria monocytogenes to stainless steel

Listeria monocytogenes ATCC 19111 cultivated in nutrient-rich medium (brain heart infusion, BHI) or starved in minimal medium (10% filter sterilized pond water and 90% sterilized distilled water) were investigated for their initial attachment to austenitic stainless steel No. 4 with satin finish at 4 °C, 20 °C, 30 °C, 37 °C, or 42 °C. A droplet (10 μl) containing  10⁷ CFU/ml of L. monocytogenes suspended in BHI or minimal medium was placed on the stainless steel surface. After holding in saturated humidity for 3 h at the desired temperature the surface was washed and prepared for scanning electron microscopy (SEM). Using SEM, attachment of L. monocytogenes was determined by counting cells remaining on the surface. When L. monocytogenes cultivated in BHI were used, with the exception of the number of attached cells being lower at 42 °C than at 37 °C and 30 °C, the number of attached cells increased with increasing temperature (P < 0.05). When L. monocytogenes starved in minimal medium were used, the number of attached cells also increased with increasing attachment temperature (P < 0.05), but the number of attached cells at 42 °C was lower than that at the other temperatures. The attachment of L. monocytogenes to stainless steel surface was greater when cultivated in rich medium of BHI vs starved in the minimal medium.     

Evaluation of adhesiveness of Acinetobacter sp. Tol 5 to abiotic surfaces

The toluene-degrading bacterium Acinetobacter sp. Tol 5 shows a highly adhesive and autoagglutinating nature through cell surface nanofibers. In the present study, various aspects of the adhesiveness of Tol 5 cells were evaluated, namely, the association with biofilm formation, the specificity, and the effects of additives. During growth, Tol 5 and Pseudomonas aeruginosa PAO1 cells showed high capacities to form biofilms. However, resting PAO1 cells barely adhered to a polystyrene (PS) surface, while a large number of resting Tol 5 cells rapidly adhered. This implies that Tol 5 cells are intrinsically adhesive and that their initial attachment ability is quite high and distinguishable from their ability to form biofilm. This high adhesiveness of Tol 5 cells was considered to be nonspecific because the cells adhered to various material surfaces from hydrophobic plastic surfaces to hydrophilic glass and metal surfaces. However, Tol 5 cells were found not to be adhesive to Escherichia coli cells. Although Tol 5 cells were capable of interspecies interaction and coagglutination with E. coli cells, at the surface of cell clumps, E. coli cells localized, suggesting that they disturbed autoagglutination of Tol 5 cells. Tol 5 has a hydrophobic cell surface. However, the addition of nonionic surfactant Triton X-100 and bovine serum albumin increased the adhesion of Tol 5 cells to a PS surface, in contrast to previous reports of hydrophobic bacteria. The results highlighted the interesting features of adhesiveness of Tol 5 cells.     

Initial adhesion of Listeria monocytogenes to solid surfaces under liquid flow

Some strains of the food borne pathogen Listeria monocytogenes persist in food processing environments. The exact reason behind this phenomenon is not known, but strain differences in the ability to adhere to solid surfaces could offer an explanation. In the present work, initial adhesion of nine strains of L. monocytogenes was investigated under liquid flow at two levels of shear stress on six different surfaces using a flow chamber set-up with microscopy measurements. The surfaces tested were glass and PVC, and glass coated with beef extract, casein, and homogenised and unhomogenised milk. In addition, the effect of prior environmental stress (5% NaCl, low nutrient availability) on initial adhesion was investigated. The hydrophobicity of the investigated surfaces was determined by contact angle measurements and the surface properties of the investigated L. monocytogenes strains were determined using Microbial Adhesion To Solvents (MATS). All surfaces with the exception of PVC were found to be hydrophilic. Strain differences were found to significantly influence the initial adhesion rate (IAR) of all nine strains to all the surfaces (p < 0.05) at both low and high shear stress. Furthermore, there was a significant effect of the surfaces tested (p < 0.05) in the adhesion ability of almost all strains. The IAR was affected by flow rate (shear stress) as seen by a decrease in adhesion at high shear stress for most strains. A significant effect of interactions between strain-surface and strain-shear stress (p < 0.001) was observed but not of interactions between surface-shear stress. No correlation between surface hydrophobicity and IAR was observed. Addition of 5% NaCl during propagation resulted in a decrease in IAR whilst propagation in low nutrient media caused an increase indicating a general change in surface characteristics under these conditions. Known persisting strains did not display general better adherence.     

Consumption of raw vegetables and fruits: a risk factor for Campylobacter infections

The purpose of this study was to determine the prevalence of Campylobacter in fresh vegetables and fruits at retail level in the Netherlands, and to estimate its implications on the importance of vegetables and fruits as risk factor for campylobacteriosis.Thirteen of the 5640 vegetable and fruit samples were Campylobacter positive, resulting in a prevalence of 0.23% (95% confidence interval (Cl): 0.12-0.39%). The prevalence of packaged products (0.36%, 95% Cl: 0.17-0.66) was significantly higher than of unpackaged products (0.07; 95% Cl: 0.01-0.27). No statistical differences were found between seasons.Combining the mean prevalence found in this study with data on the consumption of vegetables and fruits, an exposure of 0.0048 campylobacters ingested per person per day in the Netherlands by transmission via vegetables and fruits, was calculated. This exposure, as input in a Beta-Poisson dose-response model, resulted in an estimated number of 5.3 × 10⁵ cases of infection with Campylobacter per year for the whole Dutch population. This constitutes the consumption of raw vegetables and fruits, especially when packaged, to be a risk factor for Campylobacter infections.     

Lactococcal 936-species phage attachment to surface of Lactococcus lactis

The interactions of the 936-species phages sk1, jj50, and 64 with the cell surface of Lactococcus lactis LM0230 were analyzed. Cell envelopes (walls + plasma membrane), cell wall, or plasma membrane from L. lactis ssp. lactis LM0230 each inactivated the phages in vitro. However, other 936-species phages kh and P008, which do not infect strain LM0230, were not inactivated by any of the subcellular fractions. Treating cell walls or plasma membrane with the cell wall hydrolase mutanolysin eliminated inactivation of phage sk1. This suggested that intact cell wall fragments were required for inactivation. A role for plasma membrane in phage sk1 inactivation was further investigated. Boiling, washing in 2 M KCl, 8 M urea, or 0.1 M Na(2)CO(3)/pH 11, or treating the plasma membrane with proteases did not reduce adsorption or inactivation of phage. Adding lipoteichoic acid or antibodies to lipoteichoic acid did not reduce inactivation of phage in a mixture with membrane, suggesting that lipoteichoic acid was not involved. Inactivation by envelopes or cell wall correlated with ejection of DNA from the phage sk1 capsid. Although calcium is required for plaque formation, it was not required for adsorption, inactivation, or ejection of phage DNA by envelopes or cell wall. The results suggest that at least for phages sk1, jj50, and 64, adsorption and phage DNA injection into the host does not require a host membrane protein or lipoteichoic acid, and that cell wall components are sufficient for these initial steps of phage infection.     

Role of O-antigen on the Escherichia coli O157:H7 cells hydrophobicity, charge and ability to attach to lettuce

Environmental factors encountered during growing and harvesting may contribute to Escherichia coli O157:H7 contamination of lettuce. Limited nutrients and extended exposure to water may cause E. coli O157:H7 to shed its O antigen. Absence of the O157-polysaccharide antigen could affect the cell's physicochemical properties (hydrophobicity and cell charge) and ultimately influence its attachment to surfaces. The objectives of this study were to evaluate the effect of the E. coli O157:H7 O-antigen on the cell's overall hydrophobicity, charge and ability to attach to cut edge and whole leaf iceberg lettuce surfaces. Three strains of E. coli O157:H7 (86-24 wild type; F-12, mutant lacking the O-antigen and pRFBE, plasmid for O157 gene reintroduced) were examined for their hydrophobicity, overall charge and ability to attach to lettuce. Overall, E. coli O157:H7 attached at higher levels to cut surfaces over whole leaf surfaces (P = 0.008) for all strains and treatments. Additionally, the strain lacking the O-antigen (F12) — attached significantly less to lettuce (P = 0.015) than the strains expressing the antigen (WT and pRFBE). Cells lacking the O antigen (strain F-12) were also significantly more hydrophobic than strains 86-24 or pRFBE (P ≤ 0.05). Surface charge differed among the strains tested (P ≤ 0.05); however, it did not appear to influence bacterial attachment to lettuce surfaces. The charge was not fully restored in the pRFBE strain (expression of O-antigen reintroduced), therefore, no conclusions can be made pertaining to the effect of charge on attachment in this study. Results indicate that E. coli O157:H7 cells which lack the O-antigen have greater hydrophobicity and attach at lower concentrations than cells expressing the O-antigen, to iceberg lettuce surfaces.     

Campylobacter and Salmonella in raw red meats in the United Kingdom: prevalence, characterization and antimicrobial resistance pattern, 2003-2005

The prevalence of Campylobacter and Salmonella was assessed in 3959 raw red meats in the UK during 2003-2005. Meats were more frequently contaminated with Campylobacter (7.2%) than with Salmonella (2.4%). Lamb and other meats (e.g. mutton, rabbit) exhibited the highest contamination from Campylobacter (12.6% and 19.8%, respectively), compared with pork (6.3%) and beef (4.9%). Pork however had the highest contamination from Salmonella (3.9%), followed by lamb (2.0%), other meats (2.0%) and beef (1.3%). Offal samples (36.6%) were more frequently contaminated with Campylobacter or Salmonella than muscle tissue (7.0%). C. jejuni predominated in all meat types. C. coli isolates were more likely to exhibit antimicrobial drug resistance, including quinolones, than C. jejuni. Salmonella typhimurium was the most frequent Salmonella serotype isolated from meats; S. typhimurium DT104/104b isolates exhibited higher rates of multiple drug resistance than other serotypes. The findings reinforce the importance of adequate cooking of meat and good hygiene to avoid cross-contamination.     

Purification and characterization of a new bacteriocin active against Campylobacter produced by Lactobacillus salivarius SMXD51

Strain SMXD51, isolated from chicken ceca and identified as Lactobacillus salivarius, produced a component that inhibits the growth of Gram-positive and Gram-negative bacteria and especially Campylobacter jejuni. The active peptide from the cell-free supernatant of Lb. salivarius SMXD51 was purified in three steps: (i) precipitation with 80% saturated ammonium sulfate, (ii) elution on a reversed phase SPE UPTI-CLEAN cartridge using different concentrations of acetonitrile, (iii) final purification by reversed phase HPLC on a C₁₈ column. The mode of action of this peptide of 5383.2 Da was identified as bactericidal, and its amino acid composition was established. This new bacteriocin SMXD51 appears potentially very useful to reduce Campylobacter in poultry prior to processing.     

Cultured C2C12 cell lines as a model for assessment of bacterial attachment to bovine primary muscle cells

The mechanisms of bacterial attachment to meat tissues need to be understood to enhance meat safety interventions. However, little is known about attachment of foodborne pathogens to meat muscle cells. In this study, attachment of six Escherichia coli and two Salmonella strains to primary bovine muscle cells and a cultured muscle cell line, C₂C₁₂, was measured, including the effect of temperature. At 37 °C, all but one strain (EC623) attached to C₂C₁₂ cells, whereas only five of eight strains (M23Sr, H10407, EC473, Sal1729a and Sal691) attached to primary cells. At 10 °C, two strains (H10407 and EC473) attached to C₂C₁₂ cells, compared to four strains (M23Sr, EC614, H10407 and Sal1729a) of primary cells. Comparing all strains at both temperatures, EC614 displayed the highest CFU per C₂C₁₂ cell (4.60 ± 2.02 CFU/muscle cell at 37 °C), whereas greater numbers of M23Sr attached per primary cell (51.88 ± 39.43 CFU/muscle cell at 37 °C). This study indicates that primary bovine muscle cells may provide a more relevant model system to study bacterial attachment to beef carcasses compared to cell lines such as C₂C₁₂.     

Evaluation of anti-Listeria meat borne Lactobacillus for biofilm formation on selected abiotic surfaces

The ability of meat borne anti-Listeria Lactobacillus to form biofilms under different in vitro conditions and on abiotic surfaces was investigated. Biofilm formation by the adhesion to polystyrene microtiter plates was determined, this being higher for Lactobacillus curvatus CRL1532 and CRL705 and Lactobacillus sakei CRL1862. The physicochemical properties of the cell surface were relatively hydrophilic and acidic in character; L. sakei CRL1862 exhibiting the strongest autoaggregation. The adhesion of lactobacilli to stainless steel (SS) and polytetrafluoroethylene (PTFE) supports at 10 °C was found to be maximal for L. sakei CRL1862 on SS after 6 days. When biofilm architecture was characterized by epifluorescence and SEM, L. sakei CRL1862 homogeneously covered the SS surface while cell clusters were observed on PTFE; the extracellular polymeric substance matrix adapted to the topography and hydrophilic/hydrophobic characteristics of each material. The feasibility of L. sakei CRL1862 to form biofilm on materials used in meat processing highlights its potential as a control strategy for Listeria monocytogenes biofilms.     

Campylobacter contamination in broiler carcasses and correlation with slaughterhouses operational hygiene inspection

This study investigates factors associated with Campylobacter contamination of broiler carcasses, using survey data collected from nine Belgian slaughterhouses in 2008 in accordance with a European Union baseline study. Campylobacter were detected in 51.9% (202/389) (95% confidence interval, 46.8%-56.9%) of broiler carcasses. Campylobacter concentration was <10 CFU/g in 49.6% of carcasses, while 20.6% were contaminated with ≥1000 CFU/g. The mean Campylobacter concentration, as calculated by maximum likelihood estimation for left-censored data, was 1.8 log₁₀ CFU/g, with a standard deviation of 1.9 log₁₀ CFU/g. There was statistically significant variation among slaughterhouses in prevalence and concentrations of Campylobacter in their sampled carcasses. Campylobacter prevalence (but not concentrations) was positively associated with increase in broilers age. Both Campylobacter prevalence and concentration were significantly higher in carcasses sampled during June and September (but not in July and August) than carcasses sampled in January. We also investigated the correlation (Spearman’s rank correlation test) between the scores of official control inspections and Campylobacter prevalence for eight out of the nine slaughterhouses. The control inspections were routinely performed by the Belgian Federal Agency for the Safety of the Food Chain, and the concluded inspection scores were used as a general numerical indicator for the status of operational hygiene and quality of management in the slaughterhouses. Ranking of slaughterhouses based on their inspection scores was statistically correlated (Spearman’s correlation coefficient = 0.857) with their ranking based on prevalence of Campylobacter. In the present study we demonstrate how the outcomes from a routine baseline survey could be coupled with other readily available data from national control authorities in order to enable a better insight over Campylobacter contamination status in broiler slaughterhouses. Findings from this work call for subsequent in-depth investigations on technical and hygiene management factors that could impact Campylobacter contamination across broiler slaughterhouses.     

Prevalence of Campylobacter species in meat, milk and other food commodities in Pakistan

A surveillance study was carried out to determine the prevalence of Campylobacter in meat, milk and other food commodities in Pakistan. Over a period of 3 years (January 2002-December 2004), a total of 1636 food samples of meat, milk and other food commodities were procured from three big cities of Pakistan (Faisalabad, Lahore and Islamabad) and were analysed. Among meat samples, the highest prevalence (48%) of Campylobacter was recorded in raw chicken meat followed by raw beef (10.9%) and raw mutton (5.1%). Among other food commodities, the highest prevalence was observed in vegetable/fruit salad (40.9%), sandwiches (32%), cheese (11%) and raw bulk milk samples (10.2%). The overall prevalence of Campylobacter was found to be 21.5%, out of which 70.6% were identified as Campylobacter (C.) jejuni and 29.4% as C. coli. The study reported that the prevalence of Campylobacter spp. was significantly higher in the food commodities, which included raw/undercooked ingredients.     

Evaluation of ISO 10272:2006 standard versus alternative enrichment and plating combinations for enumeration and detection of Campylobacter in chicken meat

In the present study, we evaluate the recommended ISO 10272:2006 versus alternative procedures for Campylobacter enumeration and enrichment in naturally contaminated chicken meat samples (n = 49). Three enrichment media were evaluated; Bolton broth, Preston broth and CampyFood broth^® (bioMérieux SA, Marcy l’Etoile, France). In addition, three selective plating agars were compared; modified charcoal cefoperazone deoxycholate agar (mCCDA), CampyFood agar^® (CFA; bioMérieux SA) and Brilliance CampyCount agar^® (BCC; Oxoid, Basingstoke, England). Direct plating on CFA provided the highest number of Campylobacter positive samples (17/49); however this was not statistically different (P > 0.05) from numbers of positive samples recovered by direct plating on mCCDA (15/49) or BCC agars (14/49). Also, there was no significant difference between Campylobacter counts on the three compared media (P > 0.05). The coloured colonies of Campylobacter on CFA and BCC were easier to record and count than those on mCCDA. Enrichment of chicken meat samples in Bolton broth for 48 h and subsequent plating on CFA provided significantly higher (P < 0.05) Campylobacter detection compared to the other broth-agar combinations. Enrichment in Preston broth for 24 h followed by plating on mCCDA gave a higher number of positive samples (20/49) than 48 h enrichment in Bolton broth and plating on mCCDA (15/49). Enrichment in Bolton broth for 48 h followed by plating on CFA recovered 35% of samples below the limit for quantifications (<10 CFU/g, n = 34), as identified by direct plating on mCCDA. Compared to the current ISO method, some alternative combinations of enrichment and agar media could provide significantly better detection and enumeration of Campylobacter in chicken meat.     

Change in attachment of Salmonella Typhimurium, Yersinia enterocolitica, and Listeria monocytogenes to pork skin and muscle after hot water and lactic acid decontamination

The attachment of Salmonella enterica subsp. enterica serovar Typhimurium, Yersinia enterocolitica, and Listeria monocytogenes to pig skin and muscle tissue decontaminated with 80 °C water or 55 °C, 1% lactic acid for 5 and 15 s was investigated. Attachment properties differed between skin and muscle surfaces. A significantly higher number of firmly attached bacteria was found on the decontaminated skin surface compared to the non-treated skin surface, both on hot water (P < 0.0001) and on lactic acid treated skin (P < 0.001). At the muscle surfaces, no such difference in attachment were shown between hot water treated surfaces and non-treated surfaces. In contrast, for lactic acid decontamination, significantly fewer bacteria attached to the treated muscle surfaces (P < 0.0001). The study did not show significant differences in surface attachment, between Salmonella, Yersinia and Listeria, which indicate that surface and environmental factors may influence attachment more than bacterial properties. A more profound location of attached bacteria at muscle compared to skin was indicated. Confocal laser scanning microscopy studies showed that bacteria located in deep tissue structures of non-decontaminated and decontaminated skin and muscle surfaces. In the latter, bacteria tended to “hide” between the muscle fibres and may be entrapped at those sites. The finding of changed attachment properties at skin after decontamination may play a role in cross- and recontamination, during subsequent meat processing.     

The effects of Campylobacter numbers in caeca on the contamination of broiler carcasses with Campylobacter

For the presence and number of Campylobacter, 18 broiler flocks were sampled over a period of 18 months. A total of 70% of the flocks were positive for Campylobacter, with higher prevalence found in summer and autumn, compared to winter and spring. Positive flocks showed contamination rates above 90%, in negative flocks this was lower, mostly below 50%. The enumeration showed a decrease in Campylobacter during processing of positive flocks. The numbers were highest in carcasses after scalding/defeathering (mean 5.9 log(10) cfu/carcass) and dropped by 0.7 log(10) cfu/carcass after chilling. A positive correlation was observed between the number of Campylobacter present in the caeca and the number of bacteria present on carcasses and cut products. When a negative flock was slaughtered after Campylobacter positive flocks, the number of positive samples was higher compared to the case when a negative flock had been slaughtered previously. C. jejuni was isolated from 73.6% of the poultry samples.