Prevalence and Characteristics of Bacillus cereus Group Isolated from Raw and Pasteurised Milk.

The elimination of spore-forming bacteria is not guaranteed by current pasteurisation processes and is a challenging problem for the dairy industry. Given that Bacillus cereus sensu lato (B. cereus group) is an important foodborne pathogen and spoiler in the dairy industry, this study aimed at evaluating the prevalence and characteristics of B. cereus group in raw and pasteurised milk samples collected in Victoria, Australia. Isolated B. cereus group were tested for antimicrobial susceptibility, biofilm formation and virulence properties. Genetic diversity was assessed using ERIC-PCR. Proteomic profiling using MALDI-TOF MS and chemical profiling using Fourier-transform infrared (FTIR) spectroscopy were also applied for clustering of the isolates. Results showed 42.3% of milk samples contained B. cereus group, with a higher contamination level for pasteurised milk. Virulence studies identified genes nheA, nheB, hblA and nheC in most isolates and cyk gene in 46% of all isolates. Antimicrobial susceptibility testing showed a high prevalence of resistance towards ampicillin, ceftriaxone and penicillin. The biofilm-forming capacity of our isolates showed that most (53.7%) had the ability to form a biofilm. Genetic profiling using ERIC-PCR placed most B. cereus group isolates from pasteurised milk in the same cluster, indicating that they probably originated from a similar source. Raw milk isolates showed greater diversity indicating various sources. FTIR spectroscopy showed high agreement with genetic profiling. In contrast, low agreement between proteomic (MALDI-TOF MS) and genetic typing was observed. The present study showed that the FTIR spectroscopy could be adopted as a rapid tool for the typing of B. cereus group. Overall, the virulence and antimicrobial resistance characteristics, together with the ability of isolates to produce biofilm, indicate the importance of B. cereus group in the Australian dairy industry.

Prevalence, antimicrobial susceptibility and virulotyping of Listeria species and Listeria monocytogenes isolated from open-air fish markets.

BACKGROUND: The aim of this study was to investigate the prevalence and characterization of Listeria species and Listeria monocytogenes isolated from raw fish and open-air fish market environments. Eight hundred and sixty two samples including raw fish and fish market environments (samples from workers' hands, workers' knives, containers and work surface) were collected from the open-air fish markets in the Northern region of Iran. RESULTS: Listeria spp. was isolated from 104/488 (21.3%) raw fish and 29/374 (7.8%) of samples from open-air fish market environment. The isolates of Listeria spp. included L. innocua (35.3%), L. monocytogenes (32.3%), L. seeligeri (18%), and L. ivanovii (14.3%). Of the 43 L. monocytogenes isolates, 31 (72.1%), 10 (23.3%) and 2 (4.7%) belonged to serovars 1/2a, 4b, and 1/2b, respectively. The inlA, inlB, inlC, inlJ, actA, hlyA, iap, plcA, and prfA virulence-associated genes were detected in almost all of the L. monocytogenes isolates. The Listeria spp. isolates showed high resistance against tetracycline (23.3%), penicillin G, and cephalothin (each 16.5%). Besides, we observed significant resistance level to tetracycline (27.9%), ampicillin (20.9%), cephalothin, penicillin G, and streptomycin (each 16.3%) in the L. monocytogenes isolates. All of the isolates were susceptible to cefotaxime, gentamicin, kanamycin, and pefloxacin. We found that tetM (25.6%), tetA (23.3%), ampC (14%), and penA (11.6%) were the most prevalent antibiotic resistance genes in the L. monocytogenes isolates. CONCLUSIONS: Recovery of potentially pathogenic L. monocytogenes from raw fish and environment of open-air fish market samples in this study is a convincing evidence for the zoonotic potential of listeriosis.

Prevalence, characterization, and antimicrobial resistance of Yersinia species and Yersinia enterocolitica isolated from raw milk in farm bulk tanks.

The aims of this study were to investigate the prevalence and to characterize and determine the antibiotic resistance of Yersinia spp. isolates from raw milk. From September 2008 to August 2010, 446 raw milk samples were obtained from farm bulk milk tanks in Varamin, Iran. Yersinia spp. were detected in 29 (6.5%) samples, out of which 23 (79.3%), 5 (17.2%), and 1 (3.4%) were isolated from cow, sheep, and goat raw milk, respectively. The most common species isolated was Yersinia enterocolitica (65.5%), followed by Yersinia frederiksenii (31%), and Yersinia kristensenii (3.4%). Of the 19 Y. enterocolitica isolates, 14 (73.7%) were grouped into bioserotype 1A/O:9, 4 (21.1%) belonged to bioserotype 1B:O8, 1 (5.3%) belonged to bioserotype 4/O:3, and 1 isolate (biotype 1A) was not typable. All the isolates of biotypes 1B and 4harbored both the ystA and ail genes. However, all the isolates of biotype 1A were only positive for the ystB gene. The tested Yersinia spp. showed the highest percentages of resistance to tetracycline (48.3%), followed by ciprofloxacin and cephalothin (each 17.2%), ampicillin (13.8%), streptomycin (6.9%), and amoxicillin and nalidixic acid (each 3.4%). All of the tested isolates demonstrated significant sensitivity to gentamicin and chloramphenicol. Recovery of potentially pathogenic Y. enterocolitica from raw milk indicates high risks of yersiniosis associated with consumption of raw milk.

Short communication: prevalence and antibiotic resistance of Staphylococcus aureus isolated from bovine clinical mastitis.

The aims of this study were to determine the prevalence and antibiotic resistance of Staphylococcus aureus isolated from bovine clinical mastitis in Varamin, Tehran Province, Iran. All of the isolated Staph. aureus were identified by morphology and culture and confirmed using the API Staph identification system (bioMérieux, Marcy-l'Étoile, France). Antibiotic resistance genes were detected by PCR with oligonucleotide primers specific for each gene. Staphylococcus aureus was recovered from 43 of 207 (20.1%) bovine clinical milk samples. Using disk diffusion, methicillin-resistant Staph. aureus was detected in 5 of 43 (11.6%) samples. The pathogen showed high resistance against penicillin G (86%) and tetracycline (76.7%). The blaZ (penicillin) (86%), tetM (tetracycline), and ermC (erythromycin) genes (39.5% each) were the most prevalent antibiotic resistance genes. The findings of this study are useful for designing specific control programs for bovine clinical mastitis caused by Staph. aureus in this region of Iran.

Prevalence and antimicrobial resistance of Listeria, Salmonella, and Yersinia species isolates in ducks and geese.

The aims of this study were to determine the prevalence and antimicrobial resistance of Listeria, Salmonella, and Yersinia spp. isolated from duck and goose intestinal contents. A total of 471 samples, including 291 duck and 180 goose intestinal contents, were purchased from wet markets between November 2008 and July 2010. Listeria, Salmonella, and Yersinia spp. were isolated from 58 (12.3%), 107 (22.7%), and 80 (17%) of the samples, respectively. It was concluded that Listeria ivanovii, Salmonella Thompson, and Yersinia enterocolitica were the predominant serovars among Listeria, Salmonella, and Yersinia spp., respectively. Moreover, resistance to tetracycline was common in Listeria (48.3%) and Salmonella spp. (63.6%), whereas 51.3% of the Yersinia spp. isolates were resistant to cephalothin. Therefore, continued surveillance of the prevalence of the pathogens and also of emerging antibiotic resistance is needed to render possible the recognition of foods that may represent risks and also ensure the effective treatment of listeriosis, salmonellosis, and yersiniosis.

Effect of Zataria multiflora Boiss. essential oil, nisin, and their combination on the production of enterotoxin C and alpha-hemolysin by Staphylococcus aureus.

The effect of different concentrations of Zataria multiflora Boiss. essential oil (EO; 0%, 0.005%, and 0.015%), nisin (0, 0.125, and 0.25 microL/mL), and their combinations on the production of staphylococcal enterotoxin C (SEC) and alpha-hemolysin (alpha-toxin) by Staphylococcus aureus at different inoculation levels (10(3), 10(4), and 10(5) cfu/mL) in brain heart infusion broth during storage at 35 degrees C for up to 43 days was evaluated. The SEC production was significantly (p < 0.05) inhibited and the hemolysis due to alpha-toxin was significantly (p < 0.05) reduced by EO concentration at levels 0.015% and 0.005%, respectively. Significant (p < 0.05) inhibitory effect of EO on SEC production at level 0.005% was observed when it was used in combination with nisin = 0.125 microL/mL. The significant (p < 0.05) synergistic effect of EO = 0.005% and nisin = 0.125 microL/mL was also observed as more reduction of hemolysis due to alpha-toxin than EO = 0.005% alone. Further, EO significantly (p < 0.05) prevented SEC production by S. aureus during the manufacturing process of a traditional Iranian white brined cheese (as a food model) even at its lowest concentration (5 microL/100 mL), in this study.

Effect of topical 1% tropicamide on Schirmer tear test results in clinically normal horses.

OBJECTIVE: To observe the effect of topical 1% tropicamide on equine tear production as measured by Schirmer I tear test. MATERIALS AND METHODS: Fourteen adult horses received one drop of 1% tropicamide ophthalmic solution in one eye and the opposite eye served as the control. The tear production in both eyes was tested at 1, 2, 4, 6, and 24 h after 1% tropicamide administration. RESULTS: Measurements made 1 h after treatment revealed a significant reduction in Schirmer tear test values in tropicamide treated eyes (P = 0.002). The observed decrease in tear production was maintained up to 4 h after treatment (P = 0.002). Although tropicamide-induced decrease in STT values was observed in the treated eyes, the contralateral eyes did not show significant changes in Schirmer tear test results. CONCLUSION: Single dose of topical 1% tropicamide resulted in statistically significant reduction in Schirmer tear test values in clinically normal horses.

Frequency, virulence genes and antimicrobial resistance of Listeria spp. isolated from bovine clinical mastitis.

The aims of this study were to determine the prevalence, characteristics and antimicrobial resistance of Listeria spp. isolated from bovine clinical mastitis in Iran. Listeria spp. were detected in 21/207 bovine mastitic milk samples from dairy farms in Iran, comprising L. monocytogenes (n=17), L. innocua (n=3) and L. ivanovii (n=1). L. monocytogenes isolates were grouped into serogroups '4b, 4d, 4e', '1/2a, 3a', '1/2b, 3b, 7' and '1/2c, 3c'; all harboured inlA, inlC and inlJ virulence genes. Listeria spp. were most frequently resistant to penicillin G (14/21 isolates, 66.7%) and tetracyclines (11/21 isolates, 52.4%).