Application of MALDI-TOF MS to Identify and Detect Antimicrobial-Resistant Streptococcus uberis Associated with Bovine Mastitis.

Streptococcus uberis is a common bovine mastitis pathogen in dairy cattle. The rapid identification and characterization of antimicrobial resistance (AMR) in S. uberis plays an important role in its diagnosis, treatment, and prevention. In this study, matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) was used to identify S. uberis and screen for potential AMR biomarkers. Streptococcus uberis strains (n = 220) associated with bovine mastitis in northern Thailand were identified using the conventional microbiological methods and compared with the results obtained from MALDI-TOF MS. Streptococcus uberis isolates were also examined for antimicrobial susceptibility using a microdilution method. Principal component analysis (PCA) and the Mann-Whitney U test were used to analyze the MALDI-TOF mass spectrum of S. uberis and determine the difference between antimicrobial-resistant and -susceptible strains. Using MALDI-TOF MS, 73.18% (161/220) of the sampled isolates were identified as S. uberis, which conformed to the identifications obtained using conventional microbiological methods and PCR. Using PCR, antimicrobial-resistant strains could not be distinguished from antimicrobial-susceptible strains for all three antimicrobial agents, i.e., tetracycline, ceftiofur, and erythromycin. The detection of spectral peaks at 7531.20 m/z and 6804.74 m/z was statistically different between tetracycline- and erythromycin-resistant and susceptible strains, respectively. This study demonstrates a proteomic approach for the diagnosis of bovine mastitis and potentially for the surveillance of AMR among bovine mastitis pathogens.

Exploring the influence of slaughterhouse type and slaughtering steps on Campylobacter jejuni contamination in chicken meat: A cluster analysis approach.

Campylobacter jejuni (C. jejuni), a foodborne pathogen, poses notable hazards to human health and has significant economic implications for poultry production. This study aimed to assess C. jejuni contamination levels in chicken carcasses from both backyard and commercial slaughterhouses in Chiang Mai province, Thailand. It also sought to examine the effects of different slaughtering practices on contamination levels and to offer evidence-based recommendations for reducing C. jejuni contamination. Through the sampling of 105 chicken carcasses and subsequent enumeration of C. jejuni, the study captured the impact of various slaughtering practices. Utilizing k-modes clustering on the observational and bacterial count data, the research identified distinct patterns of contamination, revealing higher levels in backyard operations compared to commercial ones. The application of k-modes clustering highlighted the impact of critical slaughtering practices, particularly chilling, on contamination levels. Notably, samples with the lowest bacterial counts were typically from the chilling step, a practice predominantly found in commercial facilities. This observation underpins the recommendation for backyard slaughterhouses to incorporate ice in their post-evisceration soaking process. Mimicking commercial practices, this chilling method aims to inhibit C. jejuni growth by reducing carcass temperature, thereby enhancing food safety. Furthermore, the study suggests backyard operations adopt additional measures observed in commercial settings, like segregating equipment for each slaughtering step and implementing regular cleaning protocols. These strategic interventions are pivotal in reducing contamination risks, advancing microbiological safety in poultry processing, and aligning with global food safety enhancement efforts.

Improving the efficiency of crossbred Pradu Hang Dam chicken production for meat consumption using cold plasma technology on eggs.

The Pradu Hang Dam chicken, a Thai Native Chicken (TNCs) breed, plays an important role in many regions of Thailand because of its chewiness. However, there are some challenges with Thai Native Chicken, such as low production and slow growth rates. Therefore, this research investigates the efficiency of cold plasma technology in enhancing the production and growth rates of TNCs. First, this paper presents the embryonic development and hatch of fertile (HoF) values of treated fertilized eggs. Chicken performance indices, such as feed intake, average daily gain (ADG), feed conversion ratio (FCR), and serum growth hormone measurement, were calculated to assess chicken development. Furthermore, the potential of cost reduction was evaluated by calculating return over feed cost (ROFC). Finally, the quality aspects of chicken breast meat, including color, pH value, weight loss, cooking loss, shear force, and texture profile analysis, were investigated to evaluate cold plasma technology's impact on chicken meat. The results demonstrated that the production rate of male Pradu Hang Dam chickens (53.20%) was higher than females (46.80%). Moreover, cold plasma technology did not significantly affect chicken meat quality. According to the average return over feed cost calculation, the livestock industry could reduce feeding costs by approximately 17.42% in male chickens. Therefore, cold plasma technology is beneficial to the poultry industry to improve production and growth rates and reduce costs while being safe and environmentally friendly.

Bacterial contamination of chicken meat in slaughterhouses and the associated risk factors: A nationwide study in Thailand.

Slaughterhouses are a key source of bacterial contamination in poultry meat and products, which is a major health and economic concern for several public authorities. This study aimed to quantify the non-compliance of bacterial contamination on chicken meat sampled from slaughterhouses and identify risk factors associated with the contamination. A questionnaire survey of 569 chicken slaughterhouses was undertaken and 1,707 meat samples were collected to determine the level of bacterial contamination. The proportion of the non-compliance associated with aerobic plate count [APC] (24.6%), Staphylococcus aureus (6.3%), Enterococcus spp. (24.7%), coliforms (13.5%), Escherichia coli (33.3%), and Salmonella spp. (33.4%) based on the livestock authorities' criteria was determined. Our results highlighted that the scalding process without scalding water temperature control or improper scalding increased the risk of APC (odds ratio, OR = 4.84, 95% CI: 2.72-8.61), S. aureus (OR = 2.68, 95% CI: 1.29-5.55), Enterococcus spp. (OR = 3.38, 95% CI: 2.01-5.69), coliforms (OR = 3.01, 95% CI: 1.47-6.15), and E. coli (OR = 2.69, 95% CI: 1.58-4.56) contamination on meat samples. Meat from eviscerated carcasses was more likely to be non-compliance due to contamination by E. coli (OR = 1.96, 95% CI: 1.14-3.38). Furthermore, open or semi-closed system slaughterhouses (OR = 1.79, 95% CI: 1.23-2.60) and lack of equipment for specific slaughtering areas (OR = 1.65, 95% CI: 1.04-2.61) increased the likelihood of Salmonella spp. occurrence. This is the first study of factors influencing the non-compliance of meat samples across Thailand. Authorities can use the study findings to enhance food safety strategies at the national level.

Decontamination of Pathogenic and Spoilage Bacteria on Pork and Chicken Meat by Liquid Plasma Immersion.

In this research, we aimed to reduce the bacterial loads of Salmonella Enteritidis, Salmonella Typhimurium, Escherichia coli, Campylobacter jejuni, Staphylococcus aureus, and Pseudomonas aeruginosa in pork and chicken meat with skin by applying cold plasma in a liquid state or liquid plasma. The results showed reductions in S. Enteritidis, S. Typhimurium, E. coli, and C. jejuni on the surface of pork and chicken meat after 15 min of liquid plasma treatment on days 0, 3, 7, and 10. However, the efficacy of the reduction in S. aureus was lower after day 3 of the experiment. Moreover, P. aeruginosa could not be inactivated under the same experimental conditions. The microbial decontamination with liquid plasma did not significantly reduce the microbial load, except for C. jejuni, compared with water immersion. When compared with a control group, the pH value and water activity of pork and chicken samples treated with liquid plasma were significantly different (p ≤ 0.05), with a downward trend that was similar to those of the control and water groups. Moreover, the redness (a*) and yellowness (b*) values (CIELAB) of the meat decreased. Although the liquid plasma group resulted in an increase in the lightness (L*) values of the pork samples, these values did not significantly change in the chicken samples. This study demonstrated the efficacy of liquid plasma at reducing S. Enteritidis, S. Typhimurium, E. coli, C. jejuni, and S. aureus on the surface of pork and chicken meat during three days of storage at 4-6 °C with minimal undesirable meat characteristics.

A Cutoff Determination of Real-Time Loop-Mediated Isothermal Amplification (LAMP) for End-Point Detection of Campylobacter jejuni in Chicken Meat.

Campylobacter jejuni is one of the leading causes of foodborne illness worldwide. C. jejuni is commonly found in poultry. It is the most frequent cause of contamination and thus resulting in not only public health concerns but also economic impacts. To test for this bacterial contamination in food processing plants, this study attempted to employ a simple and rapid detection assay called loop-mediated isothermal amplification (LAMP). The best cutoff value for the positive determination of C. jejuni calculated using real-time LAMP quantification cycle (Cq) was derived from the receiver operating characteristic (ROC) curve modeling. The model showed an area under curve (AUC) of 0.936 (95% Wald CI: 0.903-0.970). Based on Youden's J statistic, the optimal cutoff value which had the highest sensitivity and specificity from the model was calculated as 18.07. The LAMP assay had 96.9% sensitivity, 95.8% specificity, and 93.9 and 97.9% positive and negative predictive values, respectively, compared to a standard culture approach for C. jejuni identification. Among all non-C. jejuni strains, the LAMP assay gave each of 12.5% false-positive results to C. coli and E. coli (1 out of 8 samples). The assay can detect C. jejuni at the lowest concentration of 103 CFU/mL. Our results suggest a preliminary indicator for the application of end-point LAMP assays, such as turbidity and UV fluorescence tests, to detect C. jejuni in field operations. The LAMP assay is an alternative screening test for C. jejuni contamination in food samples. The method provides a rapid detection, which requires only 9 min with a cutoff value of Cq. We performed the extraction of DNA from pure cultures and the detection of C. jejuni using the LAMP assay within 3 h. However, we were not able to reduce the time for the process of enrichment involved in our study. Therefore, we suggest that alternative enrichment media and rapid DNA extraction methods should be considered for further study. Compared to other traditional methods, our proposed assay requires less equipment and time, which is applicable at any processing steps in the food production chain.

Antimicrobial Resistance Profiles and Genes in Streptococcus uberis Associated With Bovine Mastitis in Thailand.

Streptococcus uberis is recognized as an environmental mastitis pathogen in dairy cattle. The varied success rate of antibiotic treatment for S. uberis intramammary infection may be associated with the antimicrobial resistance (AMR) of these bacteria. This observational study aimed to analyze 228 S. uberis strains associated with bovine mastitis in northern Thailand from 2010 to 2017. AMR and AMR genes were determined by the minimum inhibitory concentration (MIC) using a microdilution method and polymerase chain reaction, respectively. The majority of S. uberis strains were resistant to tetracycline (187/228, 82.02%), followed by ceftiofur (44/228, 19.30%), and erythromycin (19/228, 8.33%). The MIC50 and MIC90 of ceftiofur in 2017 were 2-4-fold higher than those in 2010 (P < 0.01). Resistance to tetracycline and ceftiofur significantly increased between 2010 and 2017 (P < 0.05). The most common gene detected in S. uberis was tetM (199/228, 87.28%), followed by ermB (151/228, 66.23 %) and blaZ (15/228, 6.58 %). The association between tetracycline resistance and tetM detection was statistically significant (P < 0.01). The detection rates of tetM significantly increased, while the detection rates of tetO and ermB significantly decreased during 2010-2017. AMR monitoring for bovine mastitis pathogens, especially S. uberis, is necessary to understand the trend of AMR among mastitis pathogens, which can help create an AMR stewardship program for dairy farms in Thailand.

Effect of Temperature on the Expression of Classical Enterotoxin Genes among Staphylococci Associated with Bovine Mastitis.

Staphylococcal food poisoning (SFP), caused by the contamination of staphylococcal enterotoxins, is a common foodborne disease worldwide. The aims of this study were: (1) to investigate classical staphylococcal enterotoxin genes, sea, seb, sec, sed, and see, among Staphylococcus aureus and coagulase-negative staphylococci (CNS) associated with bovine mastitis; (2) to determine the effect of temperature on the expression of classical staphylococcal enterotoxin genes in staphylococci in milk. The detection of classical staphylococcal enterotoxin genes was performed using S. aureus (n = 51) and CNS (n = 47). The expression of classical enterotoxin genes, including sea, seb, sec, and see, was determined during the growth of staphylococci in milk subjected to ultra-high-temperature processing at two different temperatures: 8 °C and room temperature. Classical staphylococcal enterotoxin genes were expressed more frequently in S. aureus (35.30%) than in CNS (12.77%). The sec gene was most frequently detected in S. aureus (29.41%) and CNS (6.38%). Moreover, the expression of sea and sec was significantly higher at room temperature than at 8 °C after 16 h of incubation (p < 0.05). These results emphasize the importance of maintaining the storage temperature of milk below 8 °C to reduce the risk of SFP.

The SCCmec Types and Antimicrobial Resistance among Methicillin-Resistant Staphylococcus Species Isolated from Dogs with Superficial Pyoderma.

This study characterizes clinical methicillin-resistant staphylococcal (MRS) isolates obtained from superficial pyoderma infections in dogs. Our interest was to determine the staphylococcal cassette chromosome mec (SCCmec) type and the antimicrobial susceptibility among MRS isolates from clinical cases. Skin swabs were collected and cultured. Staphylococcus species were identified and characterized with biochemical tests and MALDI-TOF-MS and antimicrobial susceptibility testing by disk diffusion. mecA detection and staphylococcal cassette chromosome mec (SCCmec) typing were achieved by PCR. Of the 65 clinical samples, 56 (86.2%) staphylococcal infections were identified. Twelve (21%) of 56 isolates were MRS infections. All MRS isolates were multidrug resistant. The ccrC and class-C2 mec, which were SCCmec type V, were the most prevalent (66.7%) among the 12 MRS isolates. The predominant SCCmec type V was found in S. aureus, S. intermedius group, S. lentus, S. xylosus, and S. arlettae. Treatment failure is a concern with the emergence of highly resistant MRS in dogs associated with superficial pyoderma. The detection of type V SCCmec MRS has previously been reported among veterinarians and dog owners but not in Northern Thailand. These infections serve as a reminder to improve infection prevention and control measures including reducing environmental contamination and potential zoonotic exposures to MRS. In addition, educational awareness of these risks in small animal hospitals needs to be increased among veterinary hospital staff, clients, and patients.

Acquisition Risk Factors of the SCCmec IX-Methicillin-Resistant Staphylococcus aureus in Swine Production Personnel in Chiang Mai and Lamphun Provinces, Thailand.

Methicillin-resistant Staphylococcus aureus (MRSA) harboring the type-IX staphylococcal cassette chromosome mec (SCCmec) has been found in pigs and humans in Northern Thailand. However, knowledge of the prevalence and acquisition risk factors of this MRSA strain among swine production personnel (SPP) are needed. The nasal swab samples and data were collected from 202 voluntary SPP and 31 swine farms in Chiang Mai and Lamphun Provinces, Thailand in 2017. MRSA were screened and identified using mannitol salt agar, biochemical and antimicrobial susceptibility testing, multiplex PCR, and the SCCmec typing. The prevalence of MRSA was 7.9% (16/202) and 19.3% (6/31) among SPP and swine farms. All isolates were multidrug-resistant, and 55 of 59 isolates (93%) contained the type-IX SCCmec element. Data analysis indicated that education, working time, contact frequency, working solely with swine production, and personal hygiene were significantly related to MRSA acquisition (p < 0.05). The multivariate analysis revealed that pig farming experience, working days, and showering were good predictors for MRSA carriage among SPP (area under the curve (AUC) = 0.84). The biosecurity protocols and tetracycline use were significantly associated with MRSA detection in pig farms (p < 0.05). Hence, the active surveillance of MRSA and further development of local/national intervention for MRSA control are essential.

Qualitative risk assessment of transmission pathways of highly pathogenic avian influenza (HPAI) virus at live poultry markets in Dhaka city, Bangladesh.

Analysis of environmental samples obtained from the Live Poultry Markets (LPMs) of Dhaka City, Bangladesh, has revealed that the highest degree of prevalence of highly pathogenic avian influenza A (HPAI, H5N1), besides other subtypes of the LPAI virus, poses the plausible risk of transmission of these viruses between human and poultry species. The present study was conducted using the OIE risk analysis framework to assess the risk level of each pathway successively. The estimated risk parameters were integrated towards to obtain the overall risk level for each specific HPAI transmission pathway using the matrix adapted by Cristobel Zepeda accompanying other expert consultations. The relevant data obtained from published and unpublished sources, together with survey data of field observations, were used to formulate and confirm the risk pathways and their associated risks. The results revealed that the risk of the release of the HPAI virus was medium when exposure was high. Additionally, the consequence would be considered very high with a medium degree of uncertainty for all parameters. Ultimately, the overall risk for transmission was estimated as medium with a medium degree of uncertainty. The findings of this study reveal that there is a significant threat that HPAI virus transmission could occur among poultry and humans and effectively sustain within the environment of the LPMs. Our findings are primarily focused on public health considerations, the hygienic slaughter of poultry and the relevant cleaning and sanitation practices conducted in the LPMs to support evidence-based decision-making processes. The findings of the study have the potential to be used to formulate effective risk reduction measures and can be further adapted in low-resource settings without major infrastructural changes required of the LPMs. All of which would reduce the risk of HPAI virus release and further lessen the degree of exposure and transmission in established LPMs.

Prevalence, Antibiogram, and Resistance Profile of Extended-Spectrum ß-Lactamase-Producing Escherichia coli Isolates from Pig Farms in Luzon, Philippines.

This cross-sectional study was conducted to determine the prevalence, antibiogram, and resistance profile of extended-spectrum ß-lactamase-producing Escherichia coli (ESBL-EC) isolates from healthy pigs and pig farms in Luzon, Philippines. A total of 162 rectal samples from healthy finisher and breeder pigs and boot swab samples from pig houses were collected from 54 randomly selected pig farms. Bacteria were isolated and screened using MacConkey agar plate supplemented with 1 mg/L cefotaxime. Identification of bacteria and antimicrobial susceptibility test were carried out through Vitek® 2 and combined disk test. PCR amplifications were carried out in all isolates targeting blaCTX-M and its five major groupings, blaTEM, and blaSHV. The farm prevalence of ESBL-EC was 57.41% (95% confidence interval [CI] = 43.21-70.77). A total of 48 (29.63%) ESBL-EC isolates were isolated from samples that showed 14 different phenotypic multidrug resistance patterns. The prevalence of blaCTX-M gene was 91.67% (95% CI = 80.02-97.68). All major blaCTX-M-groups except blaCTX-M-25group were detected. The blaCTX-M-1 was the most prevalent blaCTX-M gene, 75.0% (95% CI = 60.40-86.36). The prevalence of blaTEM and blaSHV genes was 91.67% (95% CI = 80.02-97.68) and 60.42% (95% CI = 45.27-74.23), respectively. Coexistence of different blaCTX-M, blaTEM, and blaSHV genes was observed in 44 isolates with 20 different genotypic patterns. High prevalence, diverse antibiogram profile, and genotypic resistance pattern of ESBL-EC isolates from healthy pigs and pig farms were observed in this study that could result in possible transmission to farm workers, susceptible bacteria, and the environment.

Prevalence and distribution of blaCTX-M, blaSHV, blaTEM genes in extended- spectrum ß- lactamase- producing E. coli isolates from broiler farms in the Philippines.

BACKGROUND: Antimicrobial resistance is a worldwide problem causing serious health threats. Escherichia coli is one of the most important bacteria that causes resistance problem. These bacteria produce an enzyme called extended-spectrum ß-lactamase (ESBL) that allows it to become resistant to a wide variety of penicillins and cephalosporins. Currently, no information or published studies on ESBL-producing E.coli in broilers are available in the Philippines. This cross-sectional study was conducted to determine the prevalence and distribution of extended-spectrum ß-lactamase (ESBL)-encoding genes, blaCTX-M, blaSHV, and blaTEM, among E. coli isolates from broiler farms in Luzon, Philippines. RESULTS: Results showed a farm prevalence of 66. 67%. A total of 69 (44.23%) ESBL-producing E. coli were isolated from boot swabs and cloacal swab samples from broiler farms. All major blaCTX-M groups except blaCTX-M-25 group were identified in the isolates. The most prevalent group was blaCTX-M-1, 72.46% (CI: 60.38-82.54%), followed by blaCTX-M-2, blaCTX-M-9 group and blaCTX-M-8. The blaTEM and blaSHV genes were identified in 57.97 and 27.54% of isolates, respectively. The blaCTX-M and blaTEM were the most common gene combinations (33.33%). Coexistence of blaCTX-M types was observed in 50 (73.53%) isolates. CONCLUSION: This study shows the high prevalence, diversity of patterns and coexistence of ESBL genes in the E. coli isolates from cloacal and boot swabs from broiler farms which pose risks of possible transmission to the environment, other animals and human.

Prevalence and Antimicrobial Resistance of Salmonella Isolates from Chicken Carcasses in Retail Markets in Yangon, Myanmar.

A cross-sectional investigation was conducted concerning prevalence, antimicrobial resistance, multidrug resistance patterns, and serovar diversity of Salmonella in chicken meat sold at retail in Yangon, Myanmar. The 141 chicken meat samples were collected at 141 retail markets in the Yangon Region, Myanmar, 1 November 2014 to 31 March 2015. Information on hygienic practices (potential risk factors) was retrieved via checklists. Salmonella was isolated and identified according to International Organization for Standardization methods (ISO 6579:2002) with minor modifications. Twelve antimicrobial agents belonging to eight pharmacological groups were used for antimicrobial susceptibility testing (disk diffusion method). Salmonella was recovered from 138 (97.9%) of the 141 samples. The isolates were most frequently resistant to trimethoprim-sulfamethoxazole (70.3% of isolates), tetracycline (54.3%), streptomycin (49.3%), and ampicillin (47.1%). Resistance was also found to chloramphenicol (29.7%), amoxicillin-clavulanic acid (17.4%), ciprofloxacin (9.4%), tobramycin (8.7%), gentamicin (8%), cefazolin (7.2%), lincomycin-spectinomycin (5.8%), and norfloxacin (0.7%). Among the 138 Salmonella isolates, 72 (52.2%) were resistant to three or more antimicrobial agents. Twenty-four serovars were identified among the 138 Salmonella-positive samples; serovars Albany, Kentucky, Braenderup, and Indiana were found in 38, 11, 10, and 8% of samples, respectively. None of the potential risk factors were significantly related to Salmonella contamination of chicken carcasses. This study provides new information regarding prevalence and antimicrobial resistance and Salmonella serovar diversity in retail markets in Yangon, Myanmar.

Prevalence and antimicrobial resistance of Vibrio spp. in retail shrimps in Vietnam.

The aim of the study was to investigate the prevalence and the antimicrobial resistance patterns of Vibrio (V.) spp. isolated from retail shrimp in Hanoi, Vietnam A total of 202 shrimp samples were collected from retail markets located in ten urban districts of Hanoi. Among those, 201 (99.5%) samples were positive for Vibrio spp. The most common species detected was V parahaemolyticus (96.5%), followed by V. alginolyticus (56.4%), V. cholerae (2%) and V. vulnificus (1.5%). Multiple Vibrio spp. were found in 114 (56.4%) samples. None of the V. parahaemolyticus isolates carried the virulence-associated tdh (thermostable direct haemolysin) and trh (tdh-related haemolysin) genes. In total, 195 V. parahaemolyticus isolates, four V. cholerae isolates and three V. vulnificus isolates were tested for resistance to eight antimicrobial agents. V. parahaemolyticus isolates showed high rates of resistance against ampicillin (87.2%), while a moderate rate was observed for sulfamethoxazole/trimethoprim (18.5%) and intermediate resistance towards tetracycline (24.6%). Low resistance rates (0.5%) were recorded against both ciprofloxacin and cefalothin. Only one V. cholerae isolate with resistance to ampicillin and two V. cholerae isolates with resistance to sulfamethoxazole/trimethoprim were found. All V. vulnificus isolates were susceptible to the eight antimicrobial agents tested. However, the number of V. vulnificus and V. cholerae was small. Multi-resistant isolates were found in V. parahaemolyticus with a low frequency (16.9%). The results of this study revealed the ubiquitous nature of Vibrio spp. in shrimp at retail. To reduce the potential risk of Vibrio infections due to handling or consumption of undercooked seafood, good manufacturing practice as well as safe handling and processing should be encouraged.

Prevalence and molecular characteristics of Vibrio spp. isolated from preharvest shrimp of the North Western Province of Sri Lanka.

This study investigated the prevalence and molecular characteristics of Vibrio spp. in farmed shrimp (Penaeus monodon) in Sri Lanka. A total of 170 shrimp samples (100 g of whole shrimp each) taken from individual ponds from 54 farms were collected 1 week prior to harvest from the North Western Province of Sri Lanka. Overall, 98.1% of the farms and 95.1% of the ponds were positive for Vibrio spp. in shrimp; at the pond level, V. parahaemolyticus (91.2%) was most common, followed by V. alginolyticus (18.8%), V. cholerae non-O1/non-O139 (4.1%), and V. vulnificus (2.4%). Multiple Vibrio spp. were detected in 20.6% of the ponds. None of the V. parahaemolyticus isolates (n = 419) were positive for the virulence-associated tdh (thermostable direct hemolysin) and trh (TDH-related hemolysin) genes. V. cholerae was confirmed by the presence of ompW, and all isolates (n = 8) were negative for the cholera toxin (ctxA) gene. V. cholerae isolates were serogrouped by PCR and identified as V. cholerae non-O1/non-O139. All four V. vulnificus strains, isolated from different ponds of two geographical regions, showed pathogenic potential; they belonged to vcgC sequence type, type B 16S rRNA genotype and contained a pilF polymorphism associated with human pathogenicity. The results of this study revealed the ubiquitous nature of vibrios in farmed shrimp. To minimize the potential risk of Vibrio infections due to handling or consumption of raw or undercooked seafood products, good manufacturing practices as well as proper handling and processing should be addressed.

Seroprevalence and risk factors associated with leptospirosis in dogs.

This study was done to determine the seroprevalence and risk factors of leptospirosis in dogs. From March to September 2004, a total of 210 dogs were randomly selected from the Small Animal Hospital, Faculty of Veterinary Medicine, Chiang Mai University. Dog sera were collected from the cephalic vein and kept at -20 degrees C until submitted to the National Institute of Health for a Microscopic Agglutination Test (MAT). Risk factors were analysed using logistic regression modelling. The prevalence of Leptospira antibodies was 11% (23/210). The most prevalent Leptospira serogroups were Bataviae 5.2% (11/210), Canicola 2.4% (5/210), Australis 1.4% (3/210), Icterohaemorrhagiae 1.4% (3/210), Ballum 0.5% (1/210), Djasiman 0.5% (1/210), Javanica 0.5% (1/ 210), Mini 0.5% (1/210), and Sejroe 0.5% (1/210). Risk factors, including signalment, environment and health status, were not significantly associated with leptospirosis antibodies. However, playing in sewage, staying outdoors >50% of the time, and consumption raw meat increased the risk of leptospirosis antibodies in dogs.