Effect of expanding farmlands with domestication of animals in the vicinity of disturbed swamps and built-up farmland ponds on population dispersion and decline of locally adapted Mansonia vectors (Diptera: Culicidae).

Background and Objectives: The adaptive processes and resilience of Mansonia vectors responsible for bioindicators can change in response to climate, land use, and environmental changes. This study evaluated the effects of expanding farmlands with the domestication of animals in the vicinity of either disturbed swamps or built-up farmland ponds on the population dispersion and decline of locally adapted Mansonia faunas as a result of expanding farmlands in Thailand. Materials and Methods: Based on environmental surveys, four different geographically defined study sites were selected: I - the expanding farmlands with domestication of livestock and pet animals in the vicinity of low-lying swamp with habitat fragmentation and aquatic vegetation; II - the expanding farmlands with domestication of pet animals in the vicinity of elevated swamp with habitat destruction and aquatic vegetation; III - the expanding farmlands with domestication of livestock and pet animals in the vicinity of low-lying farmland ponds with restoration and aquatic vegetation; and IV - the expanding farmlands with domestication of pet animals in the vicinity of elevated farmland ponds with restoration and aquatic vegetation. Human landing catch collection method was used to periodically assess the species composition and abundance of Mansonia vectors. Results: Aggregated distributions and seasonal abundances of Mansonia faunas (Mansonia uniformis, Mansonia indiana, Mansonia annulifera, Mansonia annulata, Mansonia bonneae, and Mansonia dives) with variable proportions were observed at all the study sites. A decline in the population of Mansonia faunas, except for Ma. uniformis, was observed at study sites I and II. Conclusion: The anticipated effects of expanding farmlands affected the population dispersion and decline of locally adapted Mansonia faunas, thus representing a diverse assemblage of Mansonia species with different adaptations, ecological tolerances, and host exploitation strategies in life. These effects depended either on the function of disturbed swamps or on the development of farmland ponds, whether they provided a wide range of freshwater habitats, or on the domestication of animals, whether they provided animal blood meal sources.

Emergence of zoonotic Brugia pahangi parasite in Thailand.

Zoonotic Brugia pahangi parasite infections in humans have emerged over two decades in Southeast Asia (SEA), including Malaysia and Thailand. The species is commonly found in domestic cats and dogs as the natural reservoir hosts. The sporadic transmission pattern of B. pahangi zoonosis causes childhood infections in Thailand and adulthood infections in Malaysia. It is crucial to understand the vulnerability in how zoonotic B. pahangi parasite is transmitted to susceptible persons in receptive settings and the exposure to the infection under impoverished environment to which the human-vector-animal interactions are related. This acquisition of knowledge will help multiple health science professions to apply One Health approach to strengthening the capacity in diagnosis and surveillance, and hence detecting and monitoring the "lingering" zoonotic B. pahangi infections present in vulnerable populations in Thailand and elsewhere in SEA. In this review article, the authors focused on articulating the concepts of plantation-related zoonotic B. pahangi filariasis by updating current knowledge of B. pahangi life cycle, vector's life cycle and current state of research on the epidemiology and ecology of B. pahangi zoonosis.

Study of fecal glucocorticoid metabolites in captive Asian elephants in Kanchanaburi Province, Thailand.

Background and Aim: Over the past two decades, the number of elephant camps in Thailand has increased considerably, and captive elephants have become more popular within the tourism industry. Tourist activities involving elephant exhibitions and trekking potentially affect animal health and welfare. This study aimed to investigate the relationships between a novel stress biomarker, fecal glucocorticoid metabolites (fGCM), and various factors (sex, age, weather season, tourist season, and elephant usage patterns), monitoring the fGCM concentration during and after trekking activities ceased. Materials and Methods: Fecal samples of 20 captive Asian elephants from two camps in Kanchanaburi Province were collected monthly for 1 year. The fGCM concentrations were measured using enzyme immunoassay and evaluated relative to individual demography, season, and tourist trekking activity. The mean differences of fGCMs concentrations were compared by analysis of variance and t-test statistics according to data types with p<0.5. Results: Significant differences in mean fGCM concentrations were found between age categories (p=0.001), trekking and non-trekking animals (p=0.039), and during and after trekking (p=0.023). The mean fGCM concentration of elephants aged during 0-44 years (136.7 ng/g) was significantly higher than for animals over 44 years old (107.7 ng/g), and the elephant trekking group (144.9 ng/g) was significantly higher than the other group (124.7 ng/g). Within the trekking group, the mean fGCM concentrations gradually declined to 129.13 ng/g within 8 months of trekking cessation. Conclusion: Elephant's ages and activities co-influenced the variance of fGCM concentrations. In addition, permanent tourist activity, especially trekking, can increase elephant stress. This study's findings can be applied to the health status monitoring of captive elephants and result in improved animal welfare.

Prevalence and Multidrug Resistance of Salmonella in Swine Production Chain in a Central Province, Thailand.

ABSTRACT: Salmonella causes foodborne disease outbreaks worldwide and raises concerns about public health and economic losses. To determine prevalence, serovar, antimicrobial resistance patterns, and the presence of extended-spectrum ß-lactamase (ESBL) genes in a cross-sectional study, 418 total samples from feces and carcasses (from three slaughterhouses) and pork and cutting boards (from four markets) were collected in a central Thailand province in 2017 and 2018. Of the 418 samples, 272 (65.1%) were positive for Salmonella. The prevalence of Salmonella-positive samples from markets (158 of 178; 88.8%) was significantly higher than that among samples from slaughterhouses (114 of 240; 47.5%) (P < 0.05). A total of 1,030 isolates were identified; 409 were assigned to 45 serovars, with Salmonella Rissen the most common (82 of 409; 20%). Two serovars, Salmonella Cannstatt and Salmonella Braubach, were identified for the first time in Thailand in market and slaughterhouse samples, respectively. Among 180 isolates representing 19 serovars, 133 (73.9%) exhibited multidrug resistance. Screening for ESBL production revealed that 41 (10.3%) of 399 isolates were ESBL positive. The prevalence of ESBL-producing Salmonella isolates was significantly higher among the market isolates (31 of 41; 75.6%) than among the slaughterhouse isolates in (10 of 41; 24.4%) (P < 0.05). In market samples, 24 (77.4%) of 31 isolates were recovered from pork and 7 (22.6%) were recovered from cutting boards. Nine ESBL-producing isolates carried single ESBL genes, either blaTEM (4 of 41 isolates; 9.8%) or blaCTX-M (5 of 41 isolates; 12.2%), whereas 11 (26.8%) carried both blaTEM and blaCTX-M. No ESBL-producing Salmonella isolate carried the blaSHV gene. These results suggest that pigs, their flesh, and cutting boards used for processing pork could be reservoirs for widespread ESBL-producing Salmonella isolates with multidrug resistance and outbreak potential across the food chain.

Investigation of Armigeres subalbatus, a vector of zoonotic Brugia pahangi filariasis in plantation areas in Suratthani, Southern Thailand.

In recent years, children in Thailand have been infected with zoonotic Brugia pahangi. However, the local environment of rubber or oil palm plantations, which would increase their exposure to risk factors of the infection due to mosquito transmission, is unclear. The present study first sought to determine the extent to which variations in the local landscape, such as the elevated versus low-lying ecotope of rubber or oil palm plantations, in a 2-km radius of the geographically defined landscape in a rural area of Suratthani, Southern Thailand could influence the abundance of Armigeres subalbatus and its susceptibility to zoonotic filarial parasite infections compared to Mansonia, Aedes, and Culex, and Coquillettidia. Thereafter, the filarial larvae found in the infected mosquitoes were molecularly investigated. Ar. subalbatus plantation ecotype was not only found to outnumber the local mosquitoes, but was identified as the predominant species that adapted well to the elevated ecotopes of the rubber or oil palm plantations, which existed at altitudes of 60-80 m. The overall rate of zoonotic filarial parasite infections (L1, L2, or L3 larvae) of Ar. subalbatus was 2.5% (95% CI, -0.2 to 4.1), with an average L3 load of 2.3 larvae per infected Ar. subalbatus (95% CI, -0.6 to 13.0); this is because the infections were found to be concentrated in the elevated ecotopes alone. Based on filarial orthologous ß-tubulin gene-specific touchup-nested PCR and sequence analysis using 30 L3 larva clones as representatives of 9 Ar. subalbatus infectious pools, Ar. subalbatus either carried B. pahangi or Dirofilaria immitis, or both species. Such findings suggest that Ar. subalbatus might have played an imperative role in the transmission of B. pahangi in the plantation areas infested with Ar. subalbatus.

Antimicrobial Resistance and Molecular Characterization of Methicillin-Resistant Staphylococcus aureus Isolated from Slaughtered Pigs and Pork in the Central Region of Thailand.

Methicillin-resistant Staphylococcus aureus (MRSA) have been a major public health concern in humans. Among MRSA, livestock-associated (LA)-MRSA strains have always been associated with exposure to livestock or their products and have emerged in different countries globally. Although studies have identified LA-MRSA from healthy pigs and pork in Thailand, prevalence in slaughtered pigs is still unknown. In addition, there are few reports on the epidemiology and molecular characteristics of LA-MRSA in Thailand. Hence, this is the first report investigating the epidemiology and molecular characteristics of MRSA in individual slaughtered pigs and pork in Thailand. A total of 204 nasal swab and 116 retailed pork samples were collected from three slaughterhouses and four fresh markets, respectively. Individual samples were used for screening for MRSA and obtained isolates were examined for drug- resistance profiling for 12 antimicrobial agents of 10 drug classes. In addition, SCCmec typing and multi-locus sequence typing were conducted to obtain genotype profiles. MRSA were isolated from 11 and 52 nasal swab and pork samples, respectively. The prevalence was significantly higher in the pork than in the nasal swab samples (p-value < 0.05). A high prevalence of ST9-SCCmecIX and ST398-SCCmecV with high-level antimicrobial resistance from markets and slaughterhouses indicated the spreading of MRSA with these genotypes in the Thai swine processing chains and suggested the need for further investigation to determine a control.

MULTIPLE-LOCUS VARIABLE-NUMBER TANDEM REPEAT ANALYSIS OF BRUCELLA ISOLATES FROM THAILAND.

Brucellosis-induced abortion can result in significant economic loss to farm animals. Brucellosis can be transmitted to humans during slaughter of infected animals or via consumption of contaminated food products. Strain identification of Brucella isolates can reveal the route of transmission. Brucella strains were isolated from vaginal swabs of farm animal, cow milk and from human blood cultures. Multiplex PCR was used to identify Brucella species, and owing to high DNA homology among Brucella isolates, multiple-locus variable-number tandem repeat analysis (MLVA) based on the number of tandem repeats at 16 different genomic loci was used for strain identification. Multiplex PCR categorized the isolates into B. abortus (n = 7), B. melitensis (n = 37), B. suis (n = 3), and 5 of unknown Brucella spp. MLVA-16 clustering analysis differentiated the strains into various genotypes, with Brucella isolates from the same geographic region being closely related, and revealed that the Thai isolates were phylogenetically distinct from those in other countries, including within the Southeast Asian region. Thus, MLVA-16 typing has utility in epidemiological studies.

Antimicrobial Resistance, Extended-Spectrum ß-Lactamase Productivity, and Class 1 Integrons in Escherichia coli from Healthy Swine.

Administration of antimicrobials to food-producing animals increases the risk of higher antimicrobial resistance in the normal intestinal flora of these animals. The present cross-sectional study was conducted to investigate antimicrobial susceptibility and extended-spectrum ß-lactamase (ESBL)-producing strains and to characterize class 1 integrons in Escherichia coli in healthy swine in Thailand. All 122 of the tested isolates had drug-resistant phenotypes. High resistance was found to ampicillin (98.4% of isolates), chloramphenicol (95.9%), gentamicin (78.7%), streptomycin (77.9%), tetracycline (74.6%), and cefotaxime (72.1%). Fifty-four (44.3%) of the E. coli isolates were confirmed as ESBL-producing strains. Among them, blaCTX-M (45 isolates) and blaTEM (41 isolates) were detected. Of the blaCTX-M-positive E. coli isolates, 37 carried the blaCTX-M-1 cluster, 12 carried the blaCTX-M-9 cluster, and 5 carried both clusters. Sequence analysis revealed blaTEM-1, blaTEM-135, and blaTEM-175 in 38, 2, and 1 isolate, respectively. Eighty-seven (71%) of the 122isolates carried class 1 integrons, and eight distinct drug-resistance gene cassettes with seven different integron profiles were identified in 43 of these isolates. Gene cassettes were associated with resistance to aminoglycosides (aadA1, aadA2, aadA22, or aadA23), trimethoprim (dfrA5, dfrA12, or dfrA17), and lincosamide (linF). Genes encoding ß-lactamases were not found in class 1 integrons. This study is the first to report ESBL-producing E. coli with a class 1 integron carrying the linF gene cassette in swine in Thailand. Our findings confirm that swine can be a reservoir of ESBL-producing E. coli harboring class 1 integrons, which may become a potential health risk if these integrons are transmitted to humans. Intensive analyses of animal, human, and environmental isolates are needed to control the spread of ESBL-producing E. coli strains.

ELISA for brucellosis detection based on three Brucella recombinant proteins.

Control of brucellosis among farm animals, wildlife and humans require reliable diagnosis. Rose Bengal serological test (RBT) is based on lipopolysaccharide antigen of Brucella, which may cross react with other gram-negative bacteria and produce false positive result. Immunoreactive proteins, such as outer-membrane protein BP26, ribosome recycling factor protein CP24 and Brucella lumazine synthase (BLS), previously reported to be recognized by infected sheep sera, were selected for production of recombinant proteins for use in an ELISA in order to investigate immune response among goats and cows, in comparison with commercial RBT. Cut-off value for ELISA was based on the immune response of in vitro fertilized goats and cows. Goats positive for Brucella culture or by RBT were ELISA positive for either IgG or IgM against at least one recombinant protein. For animals with negative RBT, animals with positive ELISA could be detected, and 61.6% possessed ELISA values as high as in infected animals. Thus, this ELISA procedure is proposed as an alternative to RBT for screening of brucellosis in farm animals.

Thailand momentum on policy and practice in local legislation on dengue vector control.

Over a past decade, an administrative decentralization model, adopted for local administration development in Thailand, is replacing the prior centralized (top-down) command system. The change offers challenges to local governmental agencies and other public health agencies at all the ministerial, regional, and provincial levels. A public health regulatory and legislative framework for dengue vector control by local governmental agencies is a national topic of interest because dengue control program has been integrated into healthcare services at the provincial level and also has been given priority in health plans of local governmental agencies. The enabling environments of local administrations are unique, so this critical review focuses on the authority of local governmental agencies responsible for disease prevention and control and on the functioning of local legislation with respect to dengue vector control and practices.

Border malaria associated with multidrug resistance on Thailand-Myanmar and Thailand-Cambodia borders: transmission dynamic, vulnerability, and surveillance.

This systematic review elaborates the concepts and impacts of border malaria, particularly on the emergence and spread of Plasmodium falciparum and Plasmodium vivax multidrug resistance (MDR) malaria on Thailand-Myanmar and Thailand-Cambodia borders. Border malaria encompasses any complex epidemiological settings of forest-related and forest fringe-related malaria, both regularly occurring in certain transmission areas and manifesting a trend of increased incidence in transmission prone areas along these borders, as the result of interconnections of human settlements and movement activities, cross-border population migrations, ecological changes, vector population dynamics, and multidrug resistance. For regional and global perspectives, this review analyzes and synthesizes the rationales pertaining to transmission dynamics and the vulnerabilities of border malaria that constrain surveillance and control of the world's most MDR falciparum and vivax malaria on these chaotic borders.

Touchdown-touchup nested PCR for low-copy gene detection of benzimidazole-susceptible Wuchereria bancrofti with a Wolbachia endosymbiont imported by migrant carriers.

A novel, sensitive and specific touchdown-touchup nested PCR (TNPCR) technique based on two useful molecular markers, a Wuchereria bancrofti ß-tubulin gene involved in benzimidazole susceptibility and a Wolbachia ftsZ gene involved in cell division, was developed to simultaneously detect the parasite W. bancrofti (W1) with its Wolbachia endosymbiont (W2) from both microfilaremic and post-treatment samples of at-risk migrant carriers infected with geographical W. bancrofti isolates. The detection and characterization of authentically low-copy gene-derived amplicons revealed no false positive identifications in amicrofilaremia with or without antigenemia. The W1-TNPCR was 100-fold more sensitive than the W2-TNPCR regardless of the microfilarial DNA isolation method and compared well with the thick blood film and membrane filtration techniques. These locus-specific TNPCRs could also detect Wolbachia-carrying W. bancrofti genotype in addition to a link to benzimidazole sensitivity among those with unknown infection origins that exhibited microfilaremia responsiveness against treatment with diethylcarbamazine plus albendazole. These TNPCR methods can augment the results of microscopic detection of the parasite because these methods enhance DNA isolation and PCR amplification capabilities.

Identification of Entamoeba histolytica and Entamoeba dispar by PCR assay of fecal specimens obtained from Thai/Myanmar border region.

Due to the indistinguishable morphology between Entamoeba histolytica (pathogenic) and Entamoeba dispar (non pathogenic), PCR-based assays were conducted. Based on microscopy, suspected Entamoeba cells were detected in 30 out of 455 fecal samples obtained from individuals residing at Thai/Myanmar border region. The target genes for PCR amplification included genes encoding small subunit rRNA (SSU-rRNA), chitinase and serine rich Entamoeba protein. PCR primers derived from SSU-rRNA gene amplified both E. histolytica and E. dispar genes producing an amplicon of 1,080 bp, and detected 3 out of 30 samples. PCR primers derived from chitinase gene of E. histolytica generating amplicons of 500 and 1,260 bp, samples were positive in 12 out of 30 samples. Due the large difference of gene encoding serine rich protein between E. histolytica and E. dispar, two specific sets of primers were designed. SREH-primer set, specific for E. histolytica, generated amplicons of 550 and 700 bp and detected 22 out of 30 samples. SED-primer set, specific to E. dispar, produced an amplicon of 550 bp, and together with a nested primer pair generating an amplicon of 477 bp, detected 16 out of 30 samples. Thus, detection of single and mixed infections of the two Entamoeba species could be effectively achieved directly from DNA extracted from feces without the need to culture the parasites.