Effect of Presurgical Iodine-Based Disinfection on Bacterial Colonization of the Equine Peripodal Region.

OBJECTIVE: To compare bacterial colonization after diluted iodine tincture or povidone iodine solution for presurgical disinfection of the equine peripodal region. STUDY DESIGN: Complete block design. ANIMALS: Five horses. METHODS: Disinfection protocols using iodine tincture or povidone iodine solutions were tested on 5 pairs (n = 10) equine front feet. Iodine tincture was applied to the left feet and povidone iodine to the right feet. Fixed surfaces of the sole, frog, hoof wall, and peripodal skin were swabbed pre-preparation (T0), after a standard pre-disinfection step (T1), after short disinfection with a 4-minute application of 0.5% iodine tincture or povidone iodine (T2), and after long disinfection with 12-hour soaking in 0.25% iodine tincture or povidone iodine (T3). Quantitative bacteriology was performed on each swab. RESULTS: The frog and sole were the most contaminated sites compared to hoof wall and skin at T0. Bacterial counts were significantly decreased at T2 for both solutions. Bacterial counts did not change significantly with iodine tincture at T3 but increased with povidone iodine compared to T2. Skin abrasions were detected on almost all feet but were subjectively more severe on iodine tincture-treated feet. CONCLUSION: Soaking for 12 hours with either iodine tincture or povidone iodine is not recommended as these solutions damaged the skin and bacterial recolonization was noted with povidone iodine. Four-minute disinfection using either iodine tincture or povidone iodine (0.5% available iodine) is appropriate for presurgical preparation of the equine peripodal region.

Characterization of persistent and transient Escherichia coli isolates recovered from clinical mastitis episodes in dairy cows.

Escherichia coli usually cause transient intramammary infections in dairy cows, but persistent intramammary infections have been observed. The objective of the study was to compare antimicrobial resistance and virulence genes found in persistent and transient E. coli isolated from clinical mastitis cases in a cohort of 91 Canadian dairy herds monitored over a 2-year period. Antimicrobial susceptibility was determined by broth microdilution and the presence of 27 virulence genes associated with extra-intestinal E. coli infections was determined by colony hybridization. Proportion of resistance in persistent E. coli ranged from 0.0% (enrofloxacin) to 27.8% (ampicillin and tetracycline). Proportion of resistance in transient E. coli ranged from 0.0% (enrofloxacin) to 16.8% (tetracycline). Odds of being classified as a persistent isolate increased by a factor of 1.6 (95% CI: 1.1, 2.4) for each aditional resistance observed (e.g. isolates resistant to four antimicrobial agents had 1.6 times higher odds of belonging to the persistent groups compared to isolates demonstrating resistance to three agents). Persistency was associated with higher odds of resistance to ampicillin (OR: 9.8, P<0.01) or cephalothin (OR: 7.6, P=0.02). Persistent isolates had 5.4 times higher odds (95% CI: 1.2, 24.0) of harboring virulence gene iroN. Similarly, persistent isolates had 8.6 times higher odds (95% CI: 2.8, 27.1) of possessing the virulence gene sitA. In conclusion, this study confirmed that persistency of intramammary E. coli isolates is associated with certain traits. Findings concerning iron-acquisition shed new light on the mechanisms of intramammary survival.

Draft Genome Sequence of Bovine Mastitis Isolate Staphylococcus agnetis CBMRN 20813338.

Presented here is a draft genome sequence for Staphylococcus agnetis CBMRN 20813338, isolated from a lactating dairy cow with subclinical mastitis. The genome is approximately 2,416 kb and has 35.79% G+C content. Analysis of the deduced open reading frame (ORF) set identified candidate virulence attributes in addition to potential molecular targets for species identification.

Biofilm formation by coagulase-negative staphylococci: impact on the efficacy of antimicrobials and disinfectants commonly used on dairy farms.

Coagulase-negative staphylococci (CNS) have traditionally been considered minor mastitis pathogens and are the bacteria most frequently isolated from intramammary infection. Previously, our laboratory demonstrated that a majority of CNS isolated from Canadian milk were able to form biofilm and this was strongly and positively associated with days in milk. Biofilms offer protection against antibiotics and disinfectants, and the presence of CNS biofilms near the end of the lactation cycle could have an impact on the prevention and recurrence of CNS infections in the next lactation cycle. The objective of this study was to investigate the effect of biofilm formation on efficacy of commonly used antibiotics and disinfectants against CNS. The minimal inhibitory concentration (MIC) and minimal biofilm eradication concentration (MBEC) of several CNS isolates were determined using microdilution method and the MBEC device, respectively. Biofilm cells were more resistant to a penicillin G/novobiocin combination and to ceftiofur than their planktonic counterparts and the increase in resistance ranged from 4× to 2048×. For the disinfectants, we determined the minimum contact time required for different teat disinfectants to eradicated planktonic cells and biofilms. The chlorhexidine-based teat disinfectants eradicated planktonic cells and biofilms within 30s. For iodine-based teat disinfectants, it took 2-10× longer to eradicate the biofilms than planktonic cells. In conclusion, CNS biofilms were less susceptible to antibiotics; however, chlorhexidine-based teat disinfectants were still effective against CNS biofilms. This reinforces the use of post-milking teat disinfectants as a preventive measure of intramammary infections.

Characterization of the ability of coagulase-negative staphylococci isolated from the milk of Canadian farms to form biofilms.

Mastitis is the most common and detrimental infection of the mammary gland in dairy cows and has a major economic impact on the production of milk and dairy products. Bacterial mastitis is caused by several pathogens, and the most frequently isolated bacterial species are coagulase-negative staphylocci (CNS). Although CNS are considered minor mastitis pathogens, the importance of CNS has increased over the years. However, the mechanism and factors involved in CNS intramammary infection are poorly studied and defined. Biofilms have been proposed as an important component in the persistence of CNS intramammary infection. Biofilms are defined as a cluster of bacteria enclosed in a self-produced matrix. The objectives of this study were to investigate the ability of CNS to form biofilms. A total of 255 mastitis-associated CNS isolates were investigated using a standard microtiter plate biofilm assay. The biofilms of some isolates were also observed by using confocal microscopy. The presence of biofilm-associated genes icaA, bap, aap, embP, fbe, and atlE was determined by PCR in the 255 isolates. The 5 dominant species assayed were Staphylococcus chromogenes (n=111), Staphylococcus simulans (n=53), Staphylococcus xylosus (n=25), Staphylococcus haemolyticus (n=15), and Staphylococcus epidermidis (n=13), and these represented 85% of the isolates. The data gathered were analyzed to identify significant links with the data deposited in the Canadian Bovine Mastitis Research Network database. Overall, Staph. xylosus is the species with the strongest ability to form biofilm, and Staph. epidermidis is the species with the lowest ability to form biofilm. Regardless of the species, the presence of icaA, bap, or the combination of multiple genes was associated with a greater ability to form biofilm. A strong relationship between the strength of a biofilm and days in milk was also noted, and CNS isolated later in the lactation cycle appeared to have a greater ability to form biofilm than those isolated earlier in the lactation cycle. In conclusion, Staph. xylosus is the species with the strongest biofilm formation ability. Furthermore, days in milk and gene combinations are predicted to be the variables with the strongest effect on biofilm formation by CNS.

Comparison of results for commercially available microbiological media plates with results for standard bacteriologic testing of bovine milk.

OBJECTIVE: To compare results for 3 commercially available microbiological media plates with those for standard bacteriologic testing of bovine milk. SAMPLE: Milk samples from postpartum cows and cows with a high somatic cell count (SCC) or clinical mastitis (CM). PROCEDURES: Sample-ready Staphylococcus culture medium (SRSC) plates were used to detect Staphylococcus aureus in milk samples obtained from postpartum cows and cows with a high SCC or CM. Rapid coliform count (RCC) plates were used to detect coliforms in milk samples obtained from cows with CM. Aerobic count (AC) plates were used to detect streptococci in CM samples. Fresh mastitic milk samples were frozen and then thawed to evaluate the effects of freezing for the SRSC and RCC plates. The effects of dilution (1:10) of samples were determined. Agreement of results between the commercially available plates and standard bacteriologic testing was evaluated. RESULTS: The ability of SRSC plates to detect S aureus in milk samples was highest with diluted samples from postpartum cows and cows with a high SCC or CM. Sensitivity of the RCC plate for detection of coliforms was highest with diluted mastitic milk samples. The AC plates had a poor positive predictive value for detection of streptococci in mastitic milk samples. Freezing increased S aureus detection. CONCLUSIONS AND CLINICAL RELEVANCE: Overall, the SRSC and RCC plates were accurate, were easy to use, and yielded results comparable to those of standard bacteriologic testing for the detection of S aureus and coliforms in bovine milk.

Proteomics, genomics, and pathway analyses of Escherichia coli and Staphylococcus aureus infected milk whey reveal molecular pathways and networks involved in mastitis.

Gram-negative and -positive bacteria elicit different response patterns by the host. The proteomic profiles of milk whey samples from cows naturally infected with Escherichia coli or Staphyloccocus aureus as compared to whey from healthy cows were determined by one-dimensional, liquid chromatography-tandem mass spectrometry (LC-MS/MS), bioinformatics processing, and pathway analyses. Since mammary epithelial cells contribute to immune responses in mammary glands, the genes of selected proteins were measured in MAC-T cells by real time quantitative PCR (qPCR) after stimulation with heat inactivated E. coli strain P4 and S. aureus strain Smith CP bacteria. A total of 173 proteins were identified including 73 proteins differentially expressed among normal, E. coli, and S. aureus treatment groups. E. coli was more effective at significantly altering the concentration of the affected proteins. The mRNA of 23 proteins out of 24 measured by qPCR was significantly altered in MAC-T cells. Pathway analyses identified top canonical pathways significantly enriched in our samples, the most significant being the acute phase response signaling pathway. Also, top networks of genes with significant associations to identified proteins were identified. Our study has demonstrated a wider proteome profile of E. coli and S. aureus mastitic milk whey, identified more low abundant defense proteins than reported before, and has linked for the first time identified proteins to several network functions and Biocarta pathways.

Regional profiling for determination of genotype diversity of mastitis-specific Staphylococcus aureus lineage in Canada by use of clumping factor A, pulsed-field gel electrophoresis, and spa typing.

One of the major concerns in global public health and the dairy industry is the emergence of host-specific virulent Staphylococcus aureus strains. The high degree of stability of the species genome renders detection of genetic microvariations difficult. Thus, approaches for the rapid tracking of specialized lineages are urgently needed. We used clumping factor A (clfA) to profile 87 bovine mastitis isolates from four regions in Canada and compared the results to those obtained by pulsed-field gel electrophoresis (PFGE) and spa typing. Twenty-five pulsotypes were obtained by PFGE with an index of discrimination of 0.91. These were assigned to six PFGE lineage groups A to F and seven spa types, including two novel ones. Group A had 48.3% of the isolates and group D had 43.7% of the isolates, while only 8% of the isolates were variable. The results of antimicrobial susceptibility testing indicated that all isolates were sensitive to methicillin and the non-beta-lactam antibiotics, while three isolates were resistant to penicillin and one isolate was resistant to tetracycline. All isolates had the clfA gene and belonged to 20 clfA repeat types with an index of discrimination of 0.90. The dominant clfA types, types X, Q, C, and Z, formed 82% and 43% of PFGE groups A and D, respectively, and had copy numbers that varied only within a narrow range of between 46 and 52 copies, implying clonal selection. The rest were variable and region specific. Furthermore, the dominant groups contained subpopulations in different regions across Canada. Sequence information confirmed the relatedness obtained by the use of clfA repeat copy numbers and other methods and further revealed the occurrence of full-repeat deletions and conserved host-specific codon-triplet position biases at 18-bp units. Thus, concordant with the results of PFGE and spa typing, clfA typing proved useful for revealing the clonal nature of the mastitis isolate lineage and for the rapid profiling of subpopulations with comparable discriminatory powers.

Evaluation of the California Mastitis Test as a precalving treatment selection tool for Holstein heifers.

The objective of this study was to evaluate the California Mastitis Test (CMT) and a portable electrical conductivity meter for diagnosing precalving intramammary infection (IMI) in Holstein heifers. A total of 428 dairy heifers from 23 dairy herds were enrolled between 6 and 12 days before the expected calving date from June 2002 to June 2003. Mammary secretions were tested by both diagnostic methods and by bacterial culture for evidence of IMI. California Mastitis Test was considered negative if the score was negative, trace or 1 and was considered positive otherwise. Two cut-off points were evaluated for milk electrical conductivity (>5 and >6.5 mS/cm). From this study, an overall proportion of 69% of heifers had precalving IMI and the overall heifer prevalence of major pathogen IMI was 16.8%. At the quarter level, sensitivity and specificity of CMT (68.9% and 68.4%, respectively) and milk conductivity >5 mS/cm (41.0% and 65.2%, respectively) or >6.5 mS/cm (25.2% and 83.3%, respectively) to identify all IMI were low. However, the heifer level sensitivity and specificity of CMT for major pathogens were 91.0% (81.5-96.6) and 27.5% (22.8-32.6), respectively. Using a cut-off point of 5 mS/cm, the heifer level sensitivity and specificity for major pathogens was 68.7% (56.2-79.4) and 44.1% (38.7-49.6), respectively. A conductivity cut-off value of 6.5 mS/cm decreased the sensitivity and increased the specificity to 53.7% (41.1-66.0) and 59.5% (54.0-64.8), respectively. California Mastitis Test and milk electrical conductivity are not good predictors of major pathogen IMI in heifers during the last 2 weeks before calving. However, the negative predictive values at quarter or heifer level were high and the heifer false negative rate was 6-14% using CMT or conductivity, respectively. Therefore, these measures could be useful for screening out heifers or quarters that are unlikely to have a major pathogen IMI.

Streptococcus dysgalactiae cellulitis and toxic shock like syndrome in a Brown Swiss cow.

An adult Brown Swiss cow was presented to the Large Animal Hospital of the Centre Hospitalier Universitaire Vétérinaire de l'Université de Montréal due to a postpartum downer cow syndrome. The animal had severe and generalized swelling of all 4 limbs and was in shock, as demonstrated by hypotension, dehydration, hypothermia, altered mental status, and abnormal blood parameters. It died rapidly, and necropsy revealed a generalized, severe cellulitis caused by Streptococcus dysgalactiae subsp. dysgalactiae, bronchopneumonia, and lesions of disseminated intravascular coagulation (DIC) in the kidneys. The portal of entry of the bacteria in the subcutaneous tissue was not found, as there was no history of skin trauma or mastitis. The pure growth of large quantities of an invasive Streptococcus species, associated with hypotension, coagulopathy, and renal failure, are supportive of streptococcal toxic shock-like syndrome in a bovine.

Effect of precalving intramammary treatment with pirlimycin in nulliparous Holstein heifers.

A clinical trial was conducted to determine whether prepartum intramammary pirlimycin reduces the proportion of nulliparous heifers with intramammary infection (IMI) during early lactation and improves milk production. Quarter milk samples were collected from 428 heifers, systematically allocated to treatment and control groups, 6 to 12 d before the expected calving date and 2 to 8 d after calving. At the prepartum visit, heifers in the treatment group (n = 219) received an infusion of pirlimycin hydrochloride in all 4 quarters; the control heifers (n = 209) received no infusions. Intramammary infection was detected in 69% of the heifers and 33% of the quarters before calving. After calving, the proportion of treated heifers with IMI was significantly lower than the proportion of control heifers (31% versus 45%). Staphylococcus aureus was isolated from 10% of the heifers and 3% of the quarters before calving. After calving, the proportion of IMIs due to S. aureus was significantly lower in the treated heifers than in the control heifers (5.6% versus 10%). Antibiotic treatment increased the percentage of cures and prevented new IMIs caused by gram-positive bacteria after calving. The incidence of new IMIs caused by gram-negative bacteria and yeast was higher among treated heifers than among control heifers. There was no overall effect of treatment on milk production, but there was a significant interaction effect of treatment and the interval between treatment and calving. An increase of 302 kg of milk was observed when antibiotic treatment was applied more than 1 wk before calving. Treatment did not affect the milk somatic cell count on the 1st 3 test days after calving.

Clinical and in vitro efficacy of amoxicillin against bacteria associated with feline skin wounds and abscesses.

A clinical trial involving 122 cats with infected skin wounds or abscesses presented to 10 veterinary clinics was conducted to evaluate the efficacy of 2 oral amoxicillin drug products (a paste and a suspension). A 2nd objective of the study was to identify bacteria involved in such infections and verify their in vitro sensitivity to amoxicillin. Samples of wound exudate were harvested at the time of presentation and submitted for aerobic and anaerobic culture. The sensitivity to amoxicillin of isolates thought to be infecting agents was tested, using a standard minimum inhibitory concentration method. Pasteuralla multocida and obligate anaerobes of the genera Prevotella, Fusobacterium, and Porphyromonas were the most frequently isolated pathogens. Overall, their in vitro susceptibility to amoxicillin was very good. Both drug products were clinically efficacious with a global success rate of 95.1% for cats administered oral amoxicillin at 11-22 mg/kg bodyweight (mean 13.8 mg/kg bodyweight) twice daily for 7 to 10 days.

Comparison of susceptibility to antimicrobials of bacterial isolates from companion animals in a veterinary diagnostic laboratory in Canada between 2 time points 10 years apart.

The susceptibility to antimicrobials of bacterial species most frequently isolated from companion animals in a veterinary teaching diagnostic laboratory was evaluated retrospectively. A significant decrease between 1990-1992 and 2002-2003 was noted in the susceptibility of dog isolates to the following antimicrobials: Escherichia coli to cephalothin (86% to 61%, P < 0.001); E. coli to ampicillin (85% to 67%, P < 0.001); Proteus spp. to ampicillin (92% to 71%, P < 0.01); coagulase-positive staphylococci (Staphylococcus aureus and Staphylococcus intermedius) to enrofloxacin (99% to 95%, P < 0.01). Significantly increased susceptibilities were also noted as follows: coagulase-positive staphylococci to erythromycin (78% to 90%, P < 0.001) and tetracycline (61% to 77%, P < 0.001). Despite a limited number of results available for cats, a significant increase in susceptibility was noted for Pseudomonas spp. to gentamicin (40% to 100%, P < 0.05) and for E. coli to tetracycline (59% to 80%, P < 0.05). Regular updates on the resistance to antimicrobials used in veterinary medicine are required.

Systemic Cryptococcus albidus infection in a Doberman Pinscher.

Cryptococcus albidus is a saprophytic, encapsulated yeast usually found in air, both outdoor and indoor, and sometimes on human skin. It is not usually considered to be a primary pathogen. Most cryptococcal infections of humans and animals are caused by Cryptococcus neoformans. Several cases of C. albidus infection have been reported in humans over the past 20 years. In the veterinary literature, 2 equine cases have been described: genital infection and mycotic keratitis. The present report is the first documented case of C. albidus systemic infection in a dog. Veterinarians and diagnosticians should be aware that C. albidus may be a potential canine pathogen.

Determination of Mycoplasma bovis susceptibilities against six antimicrobial agents using the E test method.

The objective of this study was to determine the susceptibility of Mycoplasma bovis against six antibiotics using the E test methodology. Fifty-eight isolates of M. bovis originating from 55 affected cattle were evaluated. Specimen originated from: lung tissue, synovial fluid, tracheo-bronchial wash, milk, and external or inner ear discharge. Antimicrobial agents tested were azythromycin, clindamycin, erythromycin, enrofloxacin, spectinomycin and tetracycline. The E test strips were placed on the surface of Hayflick plates on which organism suspension was spread. Plates were incubated at 35 degrees C in a candle jar for 72 h. MICs were then read by determining where the growth inhibition zone intersected with the MIC scale on the strip. M. bovis Donetta isolate was used as a control. All MICs were >256 microg/ml for erythromycin. MIC50 and MIC90 obtained for azythromycin were 3 and >256 microg/ml, respectively. MIC50 and MIC90 obtained for tetracycline were 4 and 8 microg/ml, respectively. MIC50 and MIC90 obtained for spectinomycin were 2 and >1021 microg/ml, respectively. MIC50 and MIC90 obtained for clindamycin were 0.19 and >256 microg/ml, respectively. MIC50 and MIC90 obtained for enrofloxacin were 0.19 and 0.25 microg/ml, respectively. Resistance was not associated with the specimen source except for azythromycin. M. bovis susceptibilities were easily determined by the E test which demonstrated the efficacy of enrofloxacin and the acquired resistance to tetracycline, spectinomycin, azythromycin and clindamycin.

Postprocessing in vitro digestion challenge to evaluate survival of Escherichia coli O157:H7 in fermented dry sausages.

Fermented dry sausages, inoculated with Escherichia coli O157:H7 during batter preparation, were submitted to an in vitro digestion challenge to evaluate the extent to which passage through the human gastrointestinal tract could inactivate the pathogenic cells, previously stressed by the manufacturing process. The numbers of surviving E. coli O157:H7 cells remained constant after a 1-min exposure of the finely chopped sausage to synthetic saliva or during the following 120-min exposure to synthetic gastric juice at an initial pH of 2.0. However, significant (P < or = 0.05) growth of the pathogen (1.03 to 2.16 log10 CFU/g) was observed in a subsequent 250-min exposure to a synthetic pancreatic juice at pH 8.0. In a different set of experiments, fractions from the gastric suspension were transferred into the synthetic pancreatic juice at 30-min intervals to mimic the dynamics of gastric emptying. Concurrently, the pH of the remaining gastric fluid was reduced to 3.0, 2.5, and 2.0 to simulate the gradual reacidification of the stomach contents after the initial buffering effect resulting from meal ingestion. Under these new conditions, pathogen growth during pancreatic challenge was observed for the first few fractions released from the stomach (90 min of exposure [pH 2.5]), but growth was no longer possible in the fractions submitted to the most severe gastric challenge (120 min of exposure [pH < 2.2]).

Efficacy of egg cleaning compounds on eggshells contaminated with Salmonella enterica serovar Enteritidis.

Salmonella Enteritidis infections of egg contents can be related to external contamination of the shell. In this study, the efficacy of three commercial cleaning and/or sanitizing compounds (sodium carbonate, sodium hypochlorite, and potassium hydroxide) was evaluated for bactericidal activity at pH values of 10, 11, and 12 against various concentrations (10(2), 10(4), or 10(6) CFU/ml) of Salmonella Enteritidis inoculated onto the eggshell surface. Efficacy of these chemical agents was also assessed against Salmonella Enteritidis in aqueous suspension. Our results indicated that none of the chemicals applied at the recommended manufacturer's concentrations (sodium carbonate, 36 ppm; other treatments, 200 ppm) could eliminate Salmonella Enteritidis from eggshells artificially contaminated with the highest bacterial concentrations (10(4) or 10(6) CFU/ml). Higher concentrations of each product, at least 5 to 20 times greater than recommended doses, were needed to destroy the bacteria on egg surfaces. However, at or slightly above the manufacturer's recommended use concentrations, all three formulations were effective against Salmonella Enteritidis in aqueous suspension (10(8) CFU/ml) or on eggshells contaminated with 10(2) CFU/ml. For both shell and suspension assays, inactivation of Salmonella Enteritidis occurred at lower concentrations at pH 12 than at pH 11 and 10. Contact time between chemicals and Salmonella apparently influenced bacterial inactivation. Extended contact times (2 to 10 min) reduced minimum chemical concentrations necessary to inactivate the bacteria. However, neither pH nor contact time influenced Salmonella Enteritidis inactivation when the initial bacterial numbers on eggshells were high.

A model study of Escherichia coli O157:H7 survival in fermented dry sausages--influence of inoculum preparation, inoculation procedure, and selected process parameters.

The influence of inoculum preparation, inoculation level, and inoculation procedure on Escherichia coli O157:H7 inactivation during the manufacture of fermented sausage was evaluated in a model study. Prior growth in glucose-enriched tryptone soya broth, which provided exposure to mildly acidic conditions (pH 4.8), had no effect on the later survival of E. coli O157: H7 strains 5-1 and ATCC 43894 under extremely acidic conditions (pH 2), but the same strains became sensitive to acidity after 7 days of incubation on the surface of refrigerated beef (as per the normal contamination route from slaughter to further processing). In subsequent sausage production trials, the extent of destruction observed for E. coli O157:H7 strains F-90, 5-1, and ATCC 43894 inoculated directly into the meat batter was unchanged when the inoculation level was decreased from 7.3 to 4.7 log CFU/g, but the level of inactivation was ca. 1 log higher when the surfaces of beef cuts, rather than the batter, were inoculated 7 days prior to processing. Regardless of processing conditions (fermentation to a pH of < or = 5.0 at 24 or 37 degrees C, drying at 14 degrees C to a water activity [a(w)] value of 0.91 or 0.79), strains F-90, 5-1, and ATCC 43894 showed similar survival capacities during the manufacture of sausage. A approximately 2-log reduction in pathogen numbers was generally obtained after samples were dried to an a(w) of 0.91, irrespective of fermentation temperature. The addition of a 5-day predrying holding stage at the fermentation temperature significantly (P < 0.05) increased pathogen inactivation when fermentation was carried out at 37 degrees C (but not when it was carried out at 24 degrees C). However, significant pathogen reductions (4 to 5 log CFU/g) were achieved only for extensively dried products (a(w) = 0.79).

Comparison of Campylobacter isolates from poultry and humans: association between in vitro virulence properties, biotypes, and pulsed-field gel electrophoresis clusters.

The in vitro virulence properties of 197 temporally and geographically related Campylobacter isolates from chicken broilers and humans were compared. Comparisons of the virulence properties associated with genotypes and biotypes were made. All isolates adhered to, and 63% invaded, INT-407 cells, whereas 13% were cytotoxic for CHO cells. CHO cell-cytotoxic extracts were also cytotoxic for INT-407 cells, but the sensitivity for Vero cells was variable. The proportion of isolates demonstrating a high invasiveness potential (>1,000 CFU ml(-1)) or Vero cell cytotoxicity was significantly higher for human than for poultry isolates. Invasiveness was associated with Campylobacter jejuni isolates of biotypes 1 and 2, whereas CHO and INT-407 cell cytotoxicity was associated with C. jejuni isolates of biotypes 3 and 4. Cytotoxic isolates were also clustered according to pulsed-field gel electrophoresis profiles.