Taeniasis among Refugees Living on Thailand-Myanmar Border, 2012.

We tested refugee camp residents on the Thailand-Myanmar border for Taenia solium infection. Taeniasis prevalence was consistent with that for other disease-endemic regions, but seropositivity indicating T. solium taeniasis was rare. Seropositivity indicating cysticercosis was 5.5% in humans, and 3.2% in pigs. Corralling pigs and providing latrines may control transmission of these tapeworms within this camp.

DISSEMINATION OF SALMONELLA ENTERICA SEQUENCE TYPES AMONG ASEAN ECONOMIC COMMUNITY COUNTRIES.

Food-borne illness caused by Salmonella enterica remains a public health problem and results in economic loss worldwide. With the up-coming establish- ment of the ASEAN Economic Community (AEC) allowing unrestricted move- ment of labor and goods, there is a higher risk of pathogen transmission among the AEC countries. This study characterized and investigated the spatial and temporal associations of S. enterica strains isolated in AEC countries during 1940- 2012 compared with those isolated in northern-Thailand during 2011-2013. Of the 173 S. enterica strains examined, 68 sequence types (STs) and 32 clonal complexes (CCs) were identified by multi loci sequence typing. Twenty-one strains belonged to four sequence types new to AEC countries, and they constituted only two CCs. A number of strains originated from various countries with multiple hosts, were highlighted. There was evidence of strains circulating in the AEC region well over a decade. Such information will be important in formulating biosecurity measures, as well as in educating regarding the risk of disease transmission in AEC.

Class 1 integrons characterization and multilocus sequence typing of Salmonella spp. from swine production chains in Chiang Mai and Lamphun provinces, Thailand.

Pigs and pork products are well known as an important source of Salmonella, one of the major zoonotic foodborne pathogens. The emergence and spread of antimicrobial resistance is becoming a major public health concern worldwide. Integrons are genetic elements known to have a role in the acquisition and expression of genes conferring antibiotic resistance. This study focuses on the prevalence of class 1 integrons-carrying Salmonella, the genetic diversity of strains of those organisms obtained from swine production chains in Chiang Mai and Lamphun provinces, Thailand, using multilocus sequence typing (MLST) and comparison of genetic diversity of sequence types of Salmonella from this study with pulsotypes identified in previous study. In 175 Salmonella strains, the overall prevalence of class 1 integrons-carrying-Salmonella was 14%. The gene cassettes array pattern "dfrA12-orfF-aadA2" was the most frequently observed. Most of the antimicrobial resistance identified was not associated with related gene cassettes harbored by Salmonella. Six sequence types were generated from 30 randomly selected strains detected by MLST. Salmonella at the human-animal-environment interface was confirmed. Linkages both in the farm to slaughterhouse contamination route and the horizontal transmission of resistance genes were demonstrated. To reduce this problem, the use of antimicrobials in livestock should be controlled by veterinarians. Education and training of food handlers as well as promotion of safe methods of food consumption are important avenues for helping prevent foodborne illness.

PREVALENCE AND ANTIMICROBIAL RESISTANCE OF SALMONELLA ISOLATED FROM CARCASSES, PROCESSING FACILITIES AND THE ENVIRONMENT SURROUNDING SMALL SCALE POULTRY SLAUGHTERHOUSES IN THAILAND.

Salmonella is a major food-borne pathogen worldwide, including Thai- land, and poultry meat plays a role as a vehicle for the spread of the disease from animals to humans. The prevalence and characteristics of Salmonella isolated from 41 small scale poultry slaughterhouses in Chiang Mai, Thailand were determined during July 2011 through May 2012. Salmonella's prevalence in live poultry, car- casses, waste water, and soil around processing plants were 3.2%, 7.3%, 22.0% and 29.0%, respectively. Eighteen different serotypes were identified, the most common being Corvallis (15.2%), followed by Rissen (13.9%), Hadar (12.7%), Enteritidis (10.1%), [I. 4,5,12:i:-] (8.8%), Stanley (8.8%), and Weltevreden (8.8%). Antimicrobial susceptibility tests revealed that 68.4% of the Salmonella spp were resistant to at least one antimicrobial while 50.6% showed multiple drug resis- tance (MDR). Specifically, 44.3% of Salmonella were resistant to nalidixic acid, followed by streptomycin (41.8%), ampicillin (34.2%), tetracycline (34.2%), and sulfamethoxazole/trimethoprim (20.3%). Salmonella contamination was found in processing lines, carcasses, and in the environment around the processing sta- tions. These findings indicate that improving hygiene management in small scale poultry slaughterhouses as well as prudent use of antimicrobial drugs is urgently needed if Salmonella contamination is to be reduced.

Distribution and characterization of methicillin-resistant Staphylococcus aureus (MRSA) at the small animal hospital, faculty of veterinary medicine, Chiang Mai University, Thailand.

Of 416 samples taken from veterinary staff (n = 30), dogs (n = 356) and various environmental sites (n = 30) at the Small Animal Hospital, Faculty of Veterinary Medicine, Chiang Mai University, Thailand, 13 samples contained methicillin-resistant Staphylococcus aureus (MRSA), of which 1 (SCCmec type II) came from veterinarian, 9 (SCCmec types I, III, IVa, V and untypeable) from dogs, and 3 (SCCmec types I, III, and IVb) from environmental samples. The MRSA isolates were 100% susceptible to vancomycin (100%), 69% to cephazolin and 62% to gentamicin, but were up to 92% resistant to tetracycline group, 69% to trimethoprim-sulfamethoxazoles and 62% to ceftriaxone. In addition, all MRSA isolates showed multidrug resistance. As the MRSA isolates from the veterinary staff and dogs were of different SCCmec types, this suggests there were no cross-infections. However, environmental contamination appears to have come from dogs, and appropriate hygienic practices should be introduced to solve this problem.

Quantification of contamination levels and particular risk of Salmonella spp. in pigs in slaughterhouses in Chiang Mai and Lamphun provinces, Thailand.

Salmonella spp. is one of the important foodborne pathogens, and the slaughtering process is recognized as a potential point of contamination and the spread of the pathogens. The three objectives of this study are first, to quantify the contamination levels of Salmonella spp. in pig skins and carcasses, second, to evaluate the outcomes from different pig supply sources and different practices at three critical steps (scalding, splitting, and washing) for Salmonella spp. contamination, and third, to assess risk of Salmonella spp. contamination in pork products after slaughtering level. The study was performed in three representative slaughterhouses in Chiang Mai and Lamphun provinces, Thailand. Investigation conducted from May 2013 through October 2013 found the overall prevalence and contamination levels mean to be 11.85% and 0.34 MPN/cm2, respectively. There was no statistically significant in Salmonella spp. prevalence and contamination levels detected with different patterns at the slaughterhouses which were supplied pigs from either co-operative or integrated farms. Factors found to reduce Salmonella spp. loads on carcasses included good practices, e.g., regular changing of water in the scalding tank after each batch and the use of chlorine in the washing step. Risk of Salmonella spp. contamination of pork products at the final stage of slaughtering was nearly 10%. Good practices and proper hygiene measures should be applied to minimize the risk of Salmonella spp. exposure in the slaughtering line, which can reduce the contamination pressure downstream at retail shops as well as for end consumers.

Assessing antimicrobial resistance profiles of Salmonella enterica in the pork production system.

Introduction. Salmonella enterica is a significant enteric pathogen affecting human and livestock health. Pork production is a common source of Salmonella contamination, with emerging multidrug resistance (MDR) posing a global health threat.Gap statement. Salmonella contamination and antimicrobial resistance (AMR) profiles in the pig production chain are underreported.Aim. To investigate the prevalence of S. enterica in the pig production chain and characterise their AMR profiles.Methodology. We collected 485 samples from pig farms, a standard pig abattoir and retail markets in Patthalung and Songkhla provinces in southern Thailand. Antimicrobial susceptibility testing was performed on these samples, and AMR profiles were determined.Results. S. enterica was detected in 68.67% of farm samples, 45.95% of abattoir samples and 50.67% of retail market samples. Analysis of 264 isolates, representing 18 serotypes, identified S. enterica serotype Rissen as the most prevalent. The predominant resistance phenotypes included ampicillin (AMP, 91.29%), tetracycline (TET, 88.26%) and streptomycin (STR, 84.47%). Over 80% of isolates showed resistance to three or more antimicrobial classes, indicating MDR. The AMP-STR-TET resistance pattern was found in nearly 70% of all MDR isolates across the production chain.Conclusions. The high prevalence of MDR is consistent with extensive antimicrobial use in the livestock sector. The presence of extensively resistant S. enterica highlights the urgent need for antimicrobial stewardship. Strengthening preventive strategies and control measures is crucial to mitigate the risk of MDR Salmonella spreading from farm to fork.

Divergent DNA methylation patterns and gene expression in MYC and CDKN2B in canine transmissible venereal tumors.

Background and Aim: Canine transmissible venereal tumor (CTVT), a unique transmissible cancer in dogs, affects the external genitalia and potentially spreads to other parts of the body. While somatic mutations in oncogenic and tumor-suppressing genes are linked to CTVT development, the impact of DNA methylation, which affects gene expression, remains unclear. This study explored whether DNA methylation in the promoter regions of the MYC oncogene and CDKN2B tumor suppressor genes in CTVTs is associated with their expression, both at the gene and protein levels. Materials and Methods: To investigate promoter DNA methylation of MYC and CDKN2B in CTVTs, we analyzed frozen tissue samples from genital CTVT (GTVTs) and extragenital CTVT (ETVTs). Genomic DNA was extracted, bisulfite-treated, and analyzed using bisulfite polymerase chain reaction (PCR) and sequencing. The messenger RNA and protein of MYC and CDKN2B were also extracted and assessed by real-time PCR and Western blotting. Matching formalin-fixed, paraffin-embedded blocks were used for immunohistochemical staining to visualize protein distribution in GTVT and ETVT tissues. Results: Although both GTVT and ETVT samples showed MYC promoter methylation, the extent of methylation differed significantly. GTVTs displayed a much higher degree of methylation, potentially explaining the more pronounced downregulation of MYC gene expression and reduction in c-MYC protein levels observed in GTVTs compared with ETVTs. Our data revealed a prevalent hypermethylation pattern in the CDKN2B promoter across both sample types. However, DNA methylation, which was expected to have a suppressive effect, did not correlate with gene/protein expression. GTVTs displayed high protein levels despite significantly reduced CDKN2B expression. Conversely, ETVTs maintained regular CDKN2B expression but exhibited reduced protein production, suggesting a complex interplay between methylation and expression in these tumors. Conclusion: MYC demonstrated a clear association between its promoter methylation status, gene expression, and protein levels; however, CDKN2B lacked this correlation, implying the involvement of methylation-independent regulatory mechanisms and highlighting the need for further investigation.

Commercial farmed swine harbour a variety of pathogenic bacteria and antimicrobial resistance genes.

Introduction. The northern region of Thailand serves as a crucial area for swine production, contributing to the Thai community food supply. Previous studies have highlighted the presence of foodborne bacterial pathogens originating from swine farms in this region, posing a threat to both human and animal health.Gap statement. Multiple swine bacterial pathogens have been studied at a species level, but the distribution and co-occurrence of bacterial pathogens in agricultural swine has not been well established.Aim. Our study employed the intestinal scraping technique to directly examine the bacterial micro-organisms interacting with the swine host.Methodology. We used shotgun metagenomic sequencing to analyse the bacterial pathogens inhabiting the caecal microbiome of swine from five commercial farms in northern Thailand.Results. A variety of pathogenic and opportunistic bacteria were identified, including Escherichia coli, Clostridium botulinum, Staphylococcus aureus and the Corynebacterium genus. From a One Health perspective, these species are important foodborne and opportunistic pathogens in both humans and agricultural animals, making swine a critical pathogen reservoir that can cause illness in humans, especially farm workers. Additionally, the swine caecal microbiome contains commensal bacteria such as Bifidobacterium, Lactobacillus and Faecalibacterium, which are associated with normal physiology and feed utilization in healthy swine. Antimicrobial resistance genes were also detected in all samples, specifically conferring resistance to tetracycline and aminoglycosides, which have historically been used extensively in swine farming.Conclusion. The findings further support the need for improved sanitation standards in swine farms, and additional monitoring of agricultural animals and farm workers to reduce contamination and improved produce safety for human consumption.

A cross-sectional study of hepatitis E virus infection in pigs in different-sized farms in northern Thailand.

Pigs are an important reservoir of hepatitis E virus (HEV) in many countries throughout the world. We evaluated the association between farm size and presence of serum antibodies against HEV, as well as other risk factors for infection in pigs raised in Nan Province, Thailand in a cross-sectional study. The sampling frame was a total-population census of all pig herds, stratified into three classes of the farm size according to criteria developed by the Nan provincial livestock health office. One-eighth of all pigs in each farm were sampled randomly. All pig-farm owners were interviewed to elicit information on general characteristics of their farms, biosecurity and hygienic procedures, and farm management. We obtained sera and fecal samples from 879 pigs to test for antibodies to HEV and HEV RNA. Odds ratios (OR) and 95% confidence intervals (CI) for risk factors for HEV seroprevalence were estimated by multivariate logistic regression. The overall prevalence of anti-HEV immunoglobulin G antibodies was 9.9%. Pigs studied from medium-sized farms had a higher HEV seroprevalence than those from larger farms (adjusted OR 4.95, 95% CI: 1.79, 13.70). Factors associated with HEV seropositivity included feeding pigs with agro-industrial byproducts, having veterinarians on farms, and presence of other pig farms within 100 m. Twenty-five (2.9%) of 875 sampled pig stools were positive for HEV RNA. Phylogenetic analysis revealed that all HEV isolates clustered to HEV genotype 3.

Colistin resistance and resistance determinants are mobile among Salmonella enterica isolates from diseased and healthy pigs in Thailand.

Salmonella is an important enteric pathogen that poses a threat to human and livestock animal health, with emerging multidrug resistance (MDR) a major public health issue globally. We investigated the prevalence of Salmonella in healthy and diseased pigs from Thai pig farms and determined their phenotypic and genotypic antimicrobial resistance profiles. A total of 150 fecal samples were collected from pigs housed in pens from four separate pig farms in southern Thailand and tested for the presence of Salmonella. Confirmed Salmonella isolates were tested for their susceptibility to 11 antimicrobials, and PCR used to detect known antimicrobial resistance genes (ARGs). Salmonella isolates were cultured from 69% (103/150) of all fecal samples, with higher prevalence in disease pigs (12/15; 80%), compared with healthy pigs (91/135; 67%). Serotype Rissen was the most frequently identified serotype among the Salmonella isolates. Resistance to ampicillin (AMP) (97%), sulfonamide-trimethoprim (SXT) (97%), and tetracycline (TET) (94%) were the most common phenotypes observed. The most common ARGs identified were blaTEM gene (99.%), tetA (87%), sul1 (77%), and dfrA1 (74%), and more than 95% of the Salmonella isolates tested were MDR - based on resistance to three or more antimicrobial classes. The most common antimicrobial resistance pattern exhibited was AMP-TET-SXT (76%), and resistance to colistin (via the mcr-1 gene) was observed in both healthy and diseased pigs. The clonal groups of PFGE analysis in serotype Typhimurium revealed the genetic relationship among Salmonella isolated from healthy and diseased pigs from different pig farms.

Campylobacter in chicken carcasses and slaughterhouses in Malaysia.

This study was conducted to determine the Campylobacter contamination rate of chicken carcasses and the processing lines of modern processing plants in Malaysia. Three hundred sixty samples were collected from 24 flocks of broiler chickens at 12 modern poultry processing plants in 6 states of Malaysia. Fresh fecal droppings were collected from crates in the arrival area. Neck skin samples were taken from processed chicken carcasses at 3 different processing stages: before inside-outside washing, after inside-outside washing and post chilling. Swab samples from the scalding tank, chilling tank and conveyer belt before chilling were also collected to determine contamination with Campylobacter in the slaughter house environment prior to slaughter. Isolation for Campylobacter was performed following ISO 10272-1:2006(E). The overall of contamination rate with Campylobacter at the 12 plants was 61.0% (220/360). Eighty point six percent of the samples from before the inside-outside wishing step were contaminated with Campylobacter, as were 62.5% of the samples after the inside washing and 38.9% after the post-chilling step. This study shows extensive contamination of chicken carcasses and slaughtering houses in Malaysia with Campylobacter.

Genome-based analysis of infrequent Salmonella serotypes through the Thai pork production chain.

Salmonella is a prevalent zoonotic foodborne pathogen. Swine and pork are implicated as important sources of salmonellosis in humans. In Chiang Mai and Lamphun Provinces in northern Thailand, there has been a high prevalence of Salmonella persistence for over a decade. Infection is usually with dominant S. enterica serotypes, including serotypes Rissen and 1,4,[5],12:i:-. However, other serotypes also contribute to disease but are less well characterized. The whole genome sequencing data of 43 S. enterica serotypes isolated from pork production chain through 2011-2014, were used to evaluate genetic diversity and ascertain the possible source of Salmonella contamination based on Core Genome Multilocus Sequence Typing (cgMLST) approach. The Salmonella serotypes recovered from farms and slaughterhouses were re-circulating by swine environmental contamination. Conversely, the Salmonella contamination in the retail market represents cross-contamination from multiple sources, including contaminated foodstuffs. Salmonella contamination in the pork production chain has the competency for host cell adhesion, host cell invasion, and intracellular survival, which is enough for the pathogenicity of salmonellosis. In addition, all of these isolates were multi-drug resistant Salmonella, which contained at least 10 antimicrobial resistance genes. This result indicated that these S. enterica serotypes also pose a significant public health risk. Our findings support the need for appropriate surveillance of food-animal products going to market to reduce public exposure to highly pathogenic, multi-drug resistant Salmonella. Acquiring information would motivate all stakeholders to reinforce sanitation standards throughout the pork production chain in order to eradicate Salmonella contamination and reduce the risk of salmonellosis in humans.

Evaluation of Bcl-2 as a marker for chronic kidney disease prediction in cats.

Chronic kidney disease (CKD) is a frequent condition in elderly cats. Bcl-2 is linked to kidney disease through the processes of apoptosis and fibrosis. The purpose of this study is to examine Bcl-2 levels in CKD and clinically healthy age-matched cats in order to evaluate the relationship between Bcl-2 levels, signalment, and blood parameters in cats with CKD. The circulating levels of Bcl-2 were determined using an immunoassay in twenty-four CKD cats and eleven clinically healthy age-matched cats by the utilization of the general linear model (GLM), Pearson correlation, principal component analysis (PCA), ROC curves, the Cox hazard model, and Kaplan-Meier survival analysis. These were all conducted in order to explore Bcl-2 levels and their connection with other variables. The Bcl-2 immunohistochemical intensity was graded in each glomerulus and tubulointerstitium. McNemar's test was performed in order to compare the expression of Bcl-2 in the two renal tissue sites. The circulating Bcl-2 of CKD cats was significantly lower than those of clinically healthy age-matched cats (P = 0.034). The presence of circulating Bcl-2 (P < 0.01) and the severity of CKD (P = 0.02) were both linked with the survival time of cats with CKD. The area under the curve (AUC) of Bcl-2 for detection of CKD was 0.723. In cats, decreased circulating Bcl-2 was associated with increased blood BUN, creatinine levels, and CKD severity. Bcl-2 protein expression was reduced in the renal tissues of CKD cats as the disease progressed, resulting in a decrease in their survival time. This study demonstrated that Bcl-2 may be effective in diagnosing feline CKD.

Transcriptome analysis of infected Crandell Rees Feline Kidney (CRFK) cells by canine parvovirus type 2c Laotian isolates.

The advent of RNA sequencing technology provides insight into the dynamic nature of tremendous transcripts within Crandell-Reese feline kidney (CRFK) cells in response to canine parvovirus (CPV-2c) infection. A total of 1,603 genes displayed differentially expressed genes (DEGs), including 789 up-regulated genes and 814 downregulated genes in the infected cells. Gene expression profiles have shown a subtle pattern of defense mechanism and immune response to CPV through significant DEGs when extensively examined via Gene Ontology (GO) and pathway analysis. Prospective GO analysis was performed and identified several enriched GO biological process terms with significant participating roles in the immune system process and defense response to virus pathway. A Gene network was constructed using the 22 most significantly enriched genes of particular interests in defense response to virus pathways to illustrate the key pathways. Eleven genes (C1QBP, CD40, HYAL2, IFNB1, IFNG, IL12B, IL6, IRF3, LSM14A, MAVS, NLRC5) were identified, which are directly related to the defense response to the virus. Results of transcriptome profiling permit us to understand the heterogeneity of DEGs during in vitro experimental study of CPV infection, reflecting a unique transcriptome signature for the CPV virus. Our findings also demonstrate a distinct scenario of enhanced CPV responses in CRFK cells for viral clearance that involved multistep and perplexity of biological processes. Collectively, our data have given a fundamental role in anti-viral immunity as our highlights of this study, thus providing outlooks on future research priorities to be important in studying CPV.

Molecular evidence for cross boundary spread of Salmonella spp. in meat sold at retail markets in the middle Mekong basin area.

BACKGROUND: The surrounding areas of the middle Mekong basin, particularly along the border between Thailand and Lao People's Democratic Republic (Lao PDR), are high-risk areas for many livestock-associated foodborne illnesses, especially salmonellosis. This study aimed to determine the prevalence and characteristics of Salmonella spp. contamination in pork, beef and chicken meats sold at retail markets in the Thailand-Laos border area surrounding the Thai-Lao Friendship Bridge I from January to May 2019. We focused on the prevalent serotypes, antimicrobial susceptibility profiles and the multilocus sequence type (MLST) genotypes of the collected Salmonella strains. RESULTS: From a total of 370 meat samples collected, 63% were positive for Salmonella, with the prevalence of 73%, 60% and 56% from pork, beef and chicken meat samples, respectively. Of all the positive samples, 53 serotypes were identified. Of these, Salmonella enterica serovar London accounted for the majority (27%), followed by serovars Corvallis (14%), and Rissen (6%). Resistance against tetracycline was found at the highest frequency (50%), followed by ampicillin (35%) and sulfamethoxazole-trimethoprim (28%). MLST revealed no evidence of shared genetic relatedness of Salmonella at retail sites among Thailand-Laos border zone. However, a diverse range of Salmonella genotypes were spread over the area. Besides, the persistence of the residential pathogen and sharing of the supply route within-country can be inferred. CONCLUSIONS: Given the high levels of contamination of retail meats, regular disinfecting of all working areas and quality control checking at pre-retail stage must be applied to reduce the transmission of Salmonella and other foodborne pathogens to consumers. The findings of this study will make a significant contribution to the current understanding of Salmonella epidemiology to enhance food security in the region.

Characterisation of Salmonella enterica clones carrying mcr-1 plasmids in meat products and patients in Northern Thailand using long read sequencing.

Salmonella spp. is an important foodborne pathogen associated with consumption of contaminated food, especially food of livestock origin. Antimicrobial resistance (AMR) in Salmonella has been reported globally and increasing AMR in food production is a major public health issue worldwide. The objective of this study was to describe the genetic relatedness among Salmonella enterica isolates, which displayed identical DNA fingerprint profiles. Ten S. enterica isolates were selected from meat and human cases with an identical rep-PCR profile of serovars Rissen (n = 4), Weltevreden (n = 4), and Stanley (n = 2). We used long-read whole genome sequencing (WGS) on the MinION sequencing platform to type isolates and investigate in silico the presence of specific AMR genes. Antimicrobial susceptibility testing was tested by disk diffusion and gradient diffusion method to corroborate the AMR phenotype. Multidrug resistance and resistance to more than one antimicrobial agent were observed in eight and nine isolates, respectively. Resistance to colistin with an accompanying mcr-1 gene was observed among the Salmonella isolates. The analysis of core genome and whole genome MLST revealed that the Salmonella from meat and human salmonellosis were genetically related. Hence, it could be concluded that meat is one of the important sources for Salmonella infection in human.

Tracking salmonella contamination in various watersheds and phenotypic and genotypic diversity.

Salmonella enterica is an important foodborne pathogen, and contamination of surface and ground water that may result from various human activities, such as animal production and urbanization, may contribute to the public health burden. The aims of this study was to determine the sources of Salmonella contamination in four different types of watersheds and to assess the relative contribution of multidrug-resistant strains. Eighty-six water samples collected from four different watershed systems, including those impacted by swine production (n = 12), residential/industrial (n = 34), crop agriculture (n = 12), and forestry (n = 28), were cultured for Salmonella and further characterized by serotyping, antimicrobial susceptibility testing, and pulsed-field gel electrophoresis genotyping. Salmonella prevalence was high in all four watersheds: residential/industrial area (58.8%), forestry (57.1%), crop agriculture (50%), and swine production (41.7%). Majority of the Salmonella isolates (87.1%) were pansusceptible. Multidrug resistance up to eight antimicrobials (R-type: AmStTeAxChCeKmGm) was detected in water samples that originated from swine production systems only. Serovars identified included Anatum, Gaminara, and Inverness (18.3% each) and Muenchen and Newport (8.7% each), Bredeny (7.6%), and Montevideo (6.8%). Pulsed-field gel electrophoresis analysis indicated genotypic relatedness among Salmonella recovered from residential/industrial and forestry-associated watersheds (genotypic cluster types A, C, D, E, F, G, H, and J), sites with relatively close geographic proximity. Swine-production-associated isolates were distinctly different from the others (genotypic cluster types B and I), corroborating the phenotypic findings. Overall, the findings suggest that all the various watersheds, including natural forest, remain important contributors of Salmonella contamination. While swine-production-associated water samples were not found to have a disproportionately high prevalence, it was the most important reservoir of multidrug-resistant strains.

Potential role of MicroRNA as a diagnostic tool in the detection of bovine mastitis.

Bovine mastitis is a major health problem that affects dairy cows and has a negative impact on milk production. The presence of microRNAs in biofluids, such as blood and milk, could play a pivotal role in the detection of bovine mastitis. The purpose of the current study was to determine the levels of microRNA gene expression in milk, in combination with other reported mastitis indicators, as a biomarker of bovine mastitis. Milk samples (n = 171) were obtained from 113 dairy cows with known disease status (i.e., healthy; n = 23 cows, subclinical mastitis; n = 45 cows, or clinical mastitis; n = 45 cows) and analyzed for the presence of MIR24-2, MIR29B-2, MIR146A, MIR148A, MIR155, MIR181A1, MIR184, and MIR223 expression using the real-time PCR (qPCR) method. The expression data were then utilized in the creation of receiver operator characteristic curves (ROC) and further analyzed by the machine learning (ML) methods. MIR29B-2, MIR146A, MIR148A, and MIR155 expression levels differed significantly among the three groups. These potential microRNA biomarkers of mastitis exhibited high sensitivity and specificity. Next, we applied ML algorithm, specifically, a decision tree (DT) model to predict the status of milk based on MIR29B-2 and MIR146A expression levels. The results suggested that MIR29B-2, when used in combination with the California mastitis test (CMT) and days in milk (DIM) data, was applicable for screening and classification of milk samples from cows as healthy, subclinical mastitis, or mastitis. MIR29B-2 appears to have sufficient discriminatory power to enable it to be utilized as a biomarker in cases where the status of a milk sample cannot be determined based on CMT results.

Detection and characterization of microRNA expression profiling and its target genes in response to canine parvovirus in Crandell Reese Feline Kidney cells.

BACKGROUND: MicroRNAs (miRNAs) play an essential role in gene regulators in many biological and molecular phenomena. Unraveling the involvement of miRNA as a key cellular factor during in vitro canine parvovirus (CPV) infection may facilitate the discovery of potential intervention candidates. However, the examination of miRNA expression profiles in CPV in tissue culture systems has not been fully elucidated. METHOD: In the present study, we utilized high-throughput small RNA-seq (sRNA-seq) technology to investigate the altered miRNA profiling in miRNA libraries from uninfected (Control) and CPV-2c infected Crandell Reese Feline Kidney cells. RESULTS: We identified five of known miRNAs (miR-222-5p, miR-365-2-5p, miR-1247-3p, miR-322-5p and miR-361-3p) and three novel miRNAs (Novel 137, Novel 141 and Novel 102) by sRNA-seq with differentially expressed genes in the miRNA repertoire of CPV-infected cells over control. We further predicted the potential target genes of the aforementioned miRNAs using sequence homology algorithms. Notably, the targets of miR-1247-3p exhibited a potential function associated with cellular defense and humoral response to CPV. To extend the probing scheme for gene targets of miR-1247-3p, we explored and performed Gene Ontology (GO) enrichment analysis of its target genes. We discovered 229 putative targets from a total of 38 enriched GO terms. The top over-represented GO enrichment in biological process were lymphocyte activation and differentiation, marginal zone B cell differentiation, negative regulation of cytokine production, negative regulation of programed cell death, and negative regulation of signaling. We next constructed a GO biological process network composed of 28 target genes of miR-1247-3p, of which, some genes, namely BCL6, DLL1, GATA3, IL6, LEF1, LFNG and WNT1 were among the genes with obviously intersected in multiple GO terms. CONCLUSION: The miRNA-1247-3p and its cognate target genes suggested their great potential as novel therapeutic targets or diagnostic biomarkers of CPV or other related viruses.