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The antimicrobial resistance (AMR) problem is one that demands urgent interventions to minimize the rapid spread in humans, animals and the environment. Extended-spectrum beta-lactamase-producing Escherichia coli (ESBL-Ec) could spread from livestock farms to environment or circulate in farms via wastewater. This study aimed to quantify and characterize ESBL-Ec in swine feces and wastewater samples collected from 2 conventional swine farms with wastewater treatment systems. Microbiological analysis was performed on fecal and wastewater samples, and the concentration of cefotaxime (CTX)-resistant bacteria, indicative of ESBL-Ec, was determined across different stages of wastewater treatment. Additionally, ESBL-Ec was isolated and identified from fresh feces, swine house wastewater, and treated wastewater from covered lagoons and photosynthetic ponds. Results indicated a higher concentration of CTX-resistant bacteria in the wastewater in the early process of wastewater treatment. However, CTXresistant bacteria were not detected in the final photosynthetic pond in one of the 2 farms. ESBL-Ec was identified in fresh feces from swine houses and wastewater treatment plants in both farms. Further antimicrobial susceptibility testing revealed diverse resistance patterns among the ESBL-Ec isolates, with tetracycline and gentamicin demonstrated the highest resistance rates. The ESBL genes identified belonged to the CTX-M-14, CTX-M-17 and CTX-M-55. This study demonstrated the presence of ESBL-Ec in swine feces and wastewater, emphasizing the importance of effective wastewater treatment before discharging to limit the spread of AMR bacteria. [ABSTRACT FROM AUTHOR]

Background and Aim: Antimicrobial resistance (AMR) is a global problem that affects human and animal health, and eggs can act as a vehicle for pathogenic and non-pathogenic resistant bacteria in the food chain. Escherichia coli is an indicator of food contamination with fecal materials as well as the occurrence and levels of AMR. This study aimed to investigate the presence of AMR, integrons, and virulence genes in E. coli isolated from eggshell samples of three egg production systems, from supermarkets in Thailand. Materials and Methods: A total of 750 hen's egg samples were purchased from supermarkets in Phayao Province: Cage eggs (250), free-range eggs (250), and organic eggs (250). Each sample was soaked in buffered peptone water (BPW), and the BPW samples were incubated at 37°C for 18-24 h. All samples were tested for E. coli by the standard conventional culture method. Then, all identified E. coli were tested for antimicrobial susceptibility to 15 antimicrobial agents by the agar disk diffusion method. All E. coli strains were subsequently found to have virulence genes and Classes 1 and 2 integrons by polymerase chain reaction. Results: Among the eggshell samples, 91 samples were identified as having E. coli (cage eggs, 24 strains; free-range eggs, 27 strains; and organic eggs, 40 strains). Then, among the E. coli strains, 47 (51.6%) were positive for at least one virulence gene. The proportion of AMR in the eggshell samples was 91.2% (83/91), and streptomycin (STR), ampicillin (AMP), and tetracycline (TET) had a high degree of resistance. Among the E. coli strains, 27 (29.7%) strains were positive for class 1 or 2 integrons, and integron-positive strains were commonly found in STR-, AMP-, and TET-resistant strains. Multidrug resistance (MDR) was detected in 57.1% (52/91) of the E. coli strains, with STR-AMP-TET (5.5%) as the most frequent pattern. The proportion of MDR in cage eggs was 75.0% (18/24), which was higher than in both free-range and organic eggs. On the other hand, 53.2% (25/47) of E. coli carrying virulence genes had MDR, distributed across the production systems as follows: Cage eggs, 76.9% (10/13); free-range eggs, 63.6% (7/11); and organic eggs, 34.8% (8/23). Conclusion: Escherichia coli was detected in eggshell samples from all three egg production systems. The high level of virulence genes, AMR, and integrons indicated the possibility of dissemination of AMR among pathogenic and commensal E. coli through eggshells. These findings could be a major concern to farmers, food handlers, and consumers, especially regarding raw egg consumption. [ABSTRACT FROM AUTHOR]

Contamination caused by microbial fecal pollution in water bodies is a serious problem in many countries, especially in low- and middle-income countries. Therefore, fecal source tracking is an important method used to understand the source of fecal contamination and to decrease the hazard of waterborne diseases that occurs in the environment. In this study, a human-associated genetic marker for Escherichia coli (H8) was used to investigate the source of fecal contamination in the Mae Klong River, Thailand. Real-time PCR was performed with this marker for 500 E. coli isolates collected from 10 sampling sites along the river, including MK10 (upstream) to MK1 (downstream). The results showed that the proportions of H8-positive isolates were 46,14,10,18,14, 38,12, 26, 32, and 14% at MK10-MK1, respectively. All positive proportions were significantly different between the locations (p < 0.001). The higher occurrence of E. coli with H8 marker detection indicated that domestic wastewater was largely discharged without proper treatment, which is attributable to the high population and the absence of proper sewage treatment in those areas. [ABSTRACT FROM AUTHOR]

Septic tank sludge or fecal sludge (FS) is often discharged on public areas without any treatment, causing environmental and adverse public health effects. Millennium Development Goal 7c (MDG 7c) targets improved access to water and sanitation because 85% of the world's total FS is often discharged on public areas. Like other countries, Thailand faces fecal slugde management (FSM) problems leading to significant fecal pathogen infections, especially in the north and northeastern regions. This study assessed the effects of unsafely-managed FS on human health risks in two cities, Natan City and Tham Lod City in Thailand, that have relatively high liver fluke and diarrhea infections, and identified the factors relating to FSM practices that affect the prevalence of liver fluke and diarrhea infections due to FS. Specific measures were proposed in delivering integrated FSM solutions and health risk protection toward the Sustainable Development Goal No. 6 (SDG6) targets of safely managed sanitation and hygiene services. Based on data collected from these cities from February to May 2019, factors relating to FSM practices that affect the prevalence of liver fluke and diarrhea infections were identified and analysed using multiple regression analysis. Based on the findings, specific measures in delivering integrated FSM solutions and health risk protection toward the Sustainable Development Goals 6 (SDG 6) are proposed. Because no FS treatment facilities operate in Natan City and Tham Lod City, almost 100% of FS is discharged directly into open drains, resulting in diarrhea and liver fluke infections among local people. The findings identified the factors relating to FSM practices that affect the prevalence of liver fluke and diarrhea infections and proposed specific measures in delivering integrated FSM solutions and health risk protection, such as promoting education programmes, avoiding direct FS discharges into open drains and nearby streams, and providing innovative FSM technologies to eradicate fecal pathogen infections. [ABSTRACT FROM AUTHOR]

Background and Aim: Bacterial contamination of drinking water is a leading cause of gastrointestinal infections. Cats may be at risk of water contamination from feces due to poor sanitation and hygiene. The objectives of the present study were to (1) evaluate the prevalence of coliform bacteria in cat drinking water and (2) identify possible risk factors leading to contamination. Materials and Methods: Fifty-five drinking water samples were collected from water containers used by cats (median age [range]: 5 years [8 months-15 years]) at their home. Using a sterile syringe, 50 mL water was collected directly from water containers. The water samples were stored in coliform enhancement media for 24 h and then submitted for bacterial culture. Results: The prevalence of fecal coliform contamination of cat drinking water was 67.27% (37/55; 95% confidence interval: 53.29-79.32%). There was no significant difference in the prevalence of coliform bacterial contamination of drinking water by age or gender of the cat or by water container type. However, bacterial contamination differed significantly between shorthaired cats and longhaired cats when comparing Escherichia coli (9/44 [20.45%] vs. 8/11 [72.72%], p<0.001) and Enterobacter spp. (16/44 [36.36%] vs. 9/11 [81.82%], p=0.007). For water that had been in a container longer than 12 h, there were significantly more contaminated tap water samples (16/19 [84.21%]) than contaminated processed water samples (9/17 [52.94%], p=0.047). Conclusion: Coliform contamination in cat drinking water is common and occurs more often in households with longhaired cats. Drinking water for cats should be changed every 12 h, especially for households using tap water. [ABSTRACT FROM AUTHOR]

The enhancement and restoration of the water quality of deteriorating surface water resources can be challenging, particularly for rivers with multiple usages, such as agriculture, animal husbandry, human residence, and industries. Recently, the performance of DNA-based microbial source tracking (MST) indicators detected by end-point and quantitative PCR assays for identifying sources of fecal pollution from human sewage, swine, and cattle and non-host-specific (universal) fecal pollution in the Tha Chin River basin, Thailand, was evaluated. The present study monitored these validated MST markers and various physicochemical and microbial water quality parameters in samples collected from twelve stations along the Tha Chin River during four sampling events in the wet and dry seasons. No significant difference in precipitation was observed between the wet and dry samplings. Universal markers (both PCR and qPCR) were detected in all 48 samples, indicating persistent and continuing fecal contamination. The sewage- and swine-specific qPCR marker concentrations did not vary among the sampling events, whereas cattle-specific qPCR markers were detected only in the wet season. Animal-specific markers were detected in the lower Tha Chin River section, which is characterized by intensive animal farming. Sewage-specific markers were also found in the lower section and near an upstream residential area. The high agreement (87.5-100%) between the PCR and qPCR results suggested that PCR could serve as a lower-cost MST screening test that requires less technical expertise. A multivariate analysis conducted using the survival analysis procedure to include censored data also emphasized the high pollution in the lower section of the river at all sampling events. Universal and swine-specific markers showed moderate correlations with microbial indicators, including total coliforms, fecal coliforms, E. coli, and enterococci. None of the MST markers or microbial parameters were associated with the measured physicochemical parameters. This study provides the first evaluation of MST markers for monitoring surface freshwater in Thailand, and the findings might aid the pollution surveillance of impaired water bodies and the development of strategies for improving their water quality. [ABSTRACT FROM AUTHOR]

Loop-mediated isothermal amplification (LAMP) has been applied for the detection of various parasites, and its application in lateral flow dipstick (LFD) can improve the convenience of point-of-care diagnosis. A novel PAR-LAMP probe and primers were designed by manual selection from a region of low variation in the ITS-2 DNA sequence. Up to six species of rumen fluke were detected by LAMP and LAMP-LFD in this study. Target specificity and sensitivity were tested, revealing a high target specificity (accuracy) and a low limit of detection (sensitivity). Different target sensitivities of paramphistome were presented, including 5 pg for Gastrothylax crumenifer and Carmyerius sp.; 1 pg for Fischoederius elongatus , Orthocoelium parvipapillatum, and O. dicranocoelium ; and 0.1 pg for Paramphistomum epiclitum. LAMP-LFD can detect a paramphistome egg even in contaminated in feces that was spiked with the egg under laboratory conditions. In addition, natural paramphistome infection in cattle from Surat Thani and Khon Kaen provinces, Thailand, was evaluated by detection of egg contamination in fecal specimens using PAR-LAMP primers. The PAR-LAMP detection result was also statistically evaluated by microscopic examination of feces. This study presents the application of novel manually designed primers in a LAMP-LFD system for improving performance in detection and diagnosis assays for paramphistomosis. [Display omitted] • Novel LAMP primers can specifically detect paramphistomes with high sensitivity. • Contamination of feces by an egg is detectable using the LAMP-LFD. • PAR-LAMP could be applied to the diagnosis of cattle based on fecal examination. [ABSTRACT FROM AUTHOR]

Aims: To evaluate the use of nonlocal, already-available strains of phages to indicate faecal contamination in Thailand waters. Methods and Results: Phages of Bacteroides fragilis strains ATCC 700786 (RYC2056PH) and ATCC 51477 (HSP40PH) were measured in 71 human and animal wastewater samples in Thailand using a double-layer agar assay. Bacteriophage RYC2056PH was detected at concentrations comparable to representative human and animal wastewater samples from European and Mediterranean countries, with 61·7 and 33·3% above the threshold value of 100 PFU 100 ml-1 in wastewater samples of human and animal origins, respectively. On the other hand, HSP40PH was detected at low concentrations in both human- and animal-polluted wastewaters. Moreover, RYC2056PH was found in 12 canal waters with human-influenced pollution and was not detected in 6 nonpolluted river waters being tested in this study. Conclusions: The presence of RYC2056PH could indicate nonsource-specific faecal contamination in Thailand. Significance and Impact of the Study: This study provided the first evidence that bacteriophages of the European-isolated B. fragilis strain RYC2056 could be used as nonsource-specific faecal indicators in the Southeast Asian region. The results of this study support the worldwide use of Bacteroides phages as faecal indicators. [ABSTRACT FROM AUTHOR]

Highlights: [•] Aedes aegypti and E. coli co-infest domestic water storage containers. [•] There were high levels of dengue vector infestation across study sites. [•] Significantly more pupae were produced in containers contaminated with E. coli. [ABSTRACT FROM AUTHOR]

In urban centres of industrialising countries, the majority of houses are served by on-site sanitation systems such as septic tanks and unsewered toilets. The faecal sludges (FS) collected from these systems are usually discharged untreated into the urban and peri-urban environment, posing great risks to water resources and to public health. Contrary to wastewater management, the development of strategies to cope with faecal sludges, adapted to the conditions prevailing in developing countries, have long been neglected. The authors describe the current situation and discuss selected issues of FS management. A proposal is made for a rational setting of sludge quality or treatment standards in economically emerging countries. The authors stipulate that regulatory setting should take into account local economic, institutional and technical conditions. Defining suitable treatment options as critical control points in securing adequate sludge quality is better than setting and relying on numerical sludge quality standards. A separate section is devoted to the practice and to regulatory aspects of (faecal) sludge use in Argentina. An overview of treatment options, which may prove sustainable in less industrialized countries is provided. Planted sludge drying beds are one of these options. It has been piloted in Thailand for four years and details on its performance and operation are presented along with data on the hygienic quality of treated biosolids. [ABSTRACT FROM AUTHOR]

• HPyV and crAssphage were highly sensitive and specific to human sewage in Thailand. • Lachno3 and BacV6-21 were less human-associated but more abundant in sewage. • CrAssphage was more abundant in sewage than HPyVs. • HPyVs were found more often in fresh- and seawater and at higher level in seawater. • HPyV and crAssphage were suggested as human-associated markers for polluted waters. Identifying sewage contamination via microbial source tracking (MST) marker genes has proven useful for effective water quality management worldwide; however, performance evaluations for these marker genes in tropical areas are limited. Therefore, this research evaluated four human-associated MST marker genes (human polyomaviruses (JC and BK viruses [HPyVs]), bacteriophage crAssphage (CPQ_056), Lachnospiraceae Lachno3, and Bacteroides BacV6-21) for tracking sewage pollution in aquatic environments of Thailand. The viral marker genes, HPyV and crAssphage were highly sensitive and specific to sewage from onsite wastewater treatment plants (OWTPs; n = 19), with no cross-detection in 120 composite swine, cattle, chicken, duck, goat, sheep, and buffalo fecal samples. The bacterial marker genes, Lachno3 and BacV6-21, demonstrated high sensitivity but moderate specificity; however, using both markers could improve specificity to >0.80 (max value of 1.00). The most abundant markers in OWTP samples were Lachno3 and BacV6-21 (5.42–8.02 and nondetect–8.05 log 10 copies/100 mL), crAssphage (5.28–7.38 log 10 copies/100 mL), and HPyVs (3.66–6.53 log 10 copies/100 mL), respectively. Due to their increased specificity, the abundance of viral markers were further investigated in environmental waters, in which HPyVs showed greater levels (up to 4.33 log 10 copies/100 mL) and greater detection rates (92.7%) in two coastal beaches (n = 41) than crAssphage (up to 3.51 log 10 copies/100 mL and 56.1%). HPyVs were also found at slightly lower levels (up to 5.10 log 10 copies/100 mL), but at higher detection rates (92.6%), in a freshwater canal (n = 27) than crAssphage (up to 5.21 log 10 copies/100 mL and 88.9%). HPyVs and crAssphage marker genes were identified as highly sensitive and specific for tracking sewage pollution in aquatic environments of Thailand. This study underlines the importance of characterizing and validating MST markers in host groups and environmental waters before including them in a water quality management toolbox. Image, graphical abstract [ABSTRACT FROM AUTHOR]

Currently, Thai livestock is rapidly expanding, especially the production of ruminants, chicken, and swine. The improper use of antibiotics will probably lead to an antimicrobial resistance problem. It has long been suspected that wastewater released from swine farms is a crucial aspect of the spread of antimicrobial resistance to the environment. Biogas systems are wastewater treatment systems commonly used on swine farms; however, little is known about the roles they play in the occurrence and transmission of resistant bacteria between biogas and non-biogas systems. This study collected pooled water, wastewater, and feces samples from five biogas farms and three non-biogas farms in Central Thailand. The samples were isolated to hemolytic E. coli (HEC) and non-hemolytic E. coli (NHEC) to test the drug resistance by using VITEK® 2 Compact (BioMérieux, USA) and detect resistant genes by using the polymerase chain reaction (PCR) technique to correlate the determined phenotypic and genotypic patterns. The results demonstrated that enumeration levels of E. coli ranged from 20.1 to 70.4 (MPN/100 ml), 105 to 107 (cfu/ml), and 105 to 109 (cfu/g), while they were 0–148.7 (MPN/100 ml), 105 to 107 (cfu/ml) and 105 to 109 (cfu/g) for water, wastewater and manure from biogas and non-biogas swine farms, respectively. The amount of E. coli in the sow feces samples was higher than the samples of nursery piglets on biogas farms at a 0.05 significant level (p < 0.05). The antimicrobial resistance indicated the relevant resistance characteristics of E. coli : the highest antimicrobial resistance was for ampicillin (AMP), followed by amoxicillin (AMX), tetracyclines (TET), chloramphenicol (C), and piperacillin (PIP), respectively. Multidrug resistance (MDR) of E. coli was 15 drugs: AMP-AMX-AMC-PIP-CEX-CEV-CPD-XNL-GM-IMP-SXT-C-TE (11.9%) and AMP-AMX-AMC-PIP-CEX-CEV-CPD-XNL-GM-IMP-SXT-C-ENR-MBR-TE (18.55%), which were the most commonly found in biogas and non-biogas swine farms, respectively. The bla TEM , tetA , sul2, and sul3 were dominantly resistant genes isolated from the water from both types of farm; while, bla TEM , aadA1 , tetA , dfrA12 , sul2 , sul3, and cmlA were isolated from feces. The amount of E. coli in the final effluent from biogas swine farms was higher than the non-biogas swine farms; however, it was not significantly different at (p > 0.05). Furthermore, the findings of study found that genotypic characteristic of HEC showed similarity 100%. Thus, it was concluded that the levels of E. coli were accelerated in biogas wastewater treatment systems, and isolated E. coli demonstrated multidrug resistance. Even though E. coli was found in different locations, it showed relevant resistance characteristics. Therefore, regular monitoring of antimicrobial resistance on livestock farms is necessary for efficient management and drug uses on farms. Image 1 • The E. coli levels isolated from biogas systems were higher than non-biogas farms for both hemolytic E. coli and non-hemolytic E. coli. • Hemolytic and non-hemolytic E. coli from both farms showed multidrug resistance in the same direction, and highest resistance toward amoxicillin, ampicillin and tetracycline. • The most resistant genes isolated from the biogas system were 11 genes, whereas, non-biogas farms were found 4 genes of bla TEM , sul2, sul3 and tetA. [ABSTRACT FROM AUTHOR]

Simple Summary: This study investigated plant diet of wild Macaca arctoides in the Malaysia–Thailand border region using a chloroplast tRNL DNA metabarcoding approach. It is a comprehensive molecular technique to assess foods eaten by primates. We have chosen chloroplast tRNL because this region has been widely used for identifying plant species. Chloroplast tRNL DNA was amplified and sequenced using the Illumina MiniSeq platform. Sequences were analyzed using the CLC Genomic Workbench software version 12.0 to check for M. arctoides plant diet. Across these samples, we successfully identified 29 plant orders, 46 families, 124 genera, and 145 species. As the first report in Malaysia, the findings provide an important understanding on diet of wild M. arctoides that only reside in Perlis State Park, Malaysia. Understanding dietary diversity is a fundamental task in the study of stump-tailed macaque, Macaca arctoides in its natural habitat. However, direct feeding observation and morphological identification using fecal samples are not effective and nearly impossible to obtain in natural habitats because this species is sensitive to human presence. As ecological methods are challenging and time-consuming, DNA metabarcoding offers a more powerful assessment of the diet. We used a chloroplast tRNL DNA metabarcoding approach to identify the diversity of plants consumed by free-ranging M. arctoides in the Malaysia–Thailand border region located in Perlis State Park, Peninsular Malaysia. DNA was extracted from three fecal samples, and chloroplast tRNL DNA was amplified and sequenced using the Illumina MiniSeq platform. Sequences were analyzed using the CLC Genomic Workbench software. A total of 145 plant species from 46 families were successfully identified as being consumed by M. arctoides. The most abundant species were yellow saraca, Saraca thaipingensis (11.70%), common fig, Ficus carica (9.33%), aramata, Clathrotropis brachypetala (5.90%), sea fig, Ficus superba (5.44%), and envireira, Malmea dielsiana (1.70%). However, Clathrotropis and Malmea are not considered Malaysian trees because of limited data available from Malaysian plant DNA. Our study is the first to identify plant taxa up to the species level consumed by stump-tailed macaques based on a DNA metabarcoding approach. This result provides an important understanding on diet of wild M. arctoides that only reside in Perlis State Park, Malaysia. [ABSTRACT FROM AUTHOR]

Background: Soil-transmitted helminth (STH) infections are major public health problems in poor and developing countries that require fecal contamination of the environment for transmission. The consumption of raw vegetables without proper washing is one of the main routes of intestinal parasite acquisition. Therefore, this study was designed to detect the prevalence of intestinal parasitic contamination in commonly consumed raw vegetables sold in three central open-air markets in Nakhon Si Thammarat province, southern Thailand.Methods: A total of 265 fresh vegetable samples consisting of peppermint, lettuce, coriander, leek, gotu kola, celery, Chinese cabbage, culantro, Thai basil, and Chinese morning glory were purchased from three central open-air markets in the Mueang, Thasala and Sichon districts from December 2016 to March 2017. Each sample was washed with physiological saline, shaken for 15 min, and then allowed to sediment. Finally, sedimentation was performed via the sedimentation concentration technique and examined using light microscopy for the detection of pathogenic parasites.Results: The overall prevalence of parasitic contamination was 35.1% (93/265). The most predominant parasite was hookworms (42.9%), followed by Strongyloides stercoralis (10.6%), Trichuris trichiura (2.6%), Ascaris lumbricoides (2.6%), and Toxocara spp. (2.6%). The highest level of contamination was found in celery, with a prevalence rate of 63.3% (19/30), while the lowest contamination level was found in Chinese morning glory, with a prevalence rate of 2.0% (2/30). The prevalence of intestinal parasite contamination in Mueang district (51.5%) was significantly higher than that in Thasala district (17.9%) and Sichon district (30.6%) (P < 0.001).Conclusion: The results of the present study demonstrate that consumption of vegetables with parasite contamination in this area represents a potential route for the transmission of parasitic infection, particularly hookworm infection. Therefore, it is necessary for health authorities to educate consumers about the proper washing of vegetables prior to consumption. Preventive methods such as wearing gloves and washing hands after handling vegetables should also be advocated to sellers who are at risk of acquiring STH infections via skin penetration. [ABSTRACT FROM AUTHOR]