Oranuch Sanpool

Background: To address the significant burden of helminthiases in Thailand, this cross-sectional study compared the performance of a fecal parasite concentrator kit (FPCK) against the Kato–Katz (KK) method for diagnosing intestinal parasites in endemic populations across the Northeast and Southern regions. Methods: Stool samples were collected from 140 participants and examined for intestinal parasitic infections using both FPCK and KK methods. Results: The FPCK method demonstrated a significantly higher detection rate of 45.0% compared to 35.0% for the KK method. For detecting liver fluke (Opisthorchis viverrini), the FPCK method detected significantly more cases than the KK method (10.71% vs. 4.29%) (p = 0.0027). For other parasites such as Trichuris trichiura, Strongyloides stercoralis, and Entamoeba coli, the FPCK method tended to detect more infections, but the differences were not statistically significant (p > 0.05). Conclusions: The FPCK method showed better performance than the KK method for detecting intestinal helminth infections in stool samples, particularly O. viverrini, T. trichiura, S. stercoralis, and Entamoeba coli. Therefore, FPCK could be used as a suitable stool examination method for surveillance and monitoring of preventive treatment for opisthorchiasis.

Background: Human cysticercosis, caused by the larval stage (cysticerci) of the pork tapeworm Taenia solium, is an important zoonotic disease. The disease is prevalent in developing countries where porcine cysticercosis is common and undercooked pork is habitually consumed. Objective: This study aimed to develop an immunochromatography-based test kit for the rapid diagnosis of human cysticercosis using low-molecular-weight antigens purified from cyst fluid of the T. solium Asian genotype to detect specific IgG antibodies in whole blood. The kit was designated as “the cysticercosis whole-blood test kit (iCys WB kit).” Methods: It was evaluated under laboratory conditions using 164 whole-blood samples, of which 21 were from confirmed cysticercosis cases. The results of the iCys WB kit, which detects anti-T. solium (cysticercus) antibodies in simulated whole blood samples, were compared with results from corresponding human serum samples. Results: When using both sample types, iCys WB kit demonstrated an accuracy of 98.8%, a sensitivity of 91.7%, a specificity of 100%, a positive likelihood ratio of 0, a negative likelihood ratio of 0.083, and an ROC area of 0.96. The agreement between results obtained from simulated whole-blood and serum samples showed perfect concordance. Conclusions: The iCys WB kit is a valuable easy-to-handle diagnostic tool and may be applicable for supporting clinical diagnosis at the point of care.

Human cysticercosis is a serious zoonosis caused by infection with larvae (cysticerci) of the pork tapeworm, Taenia solium . Infection can involve the nervous system, causing chronic headache and intracranial hypertension, focal neurological deficits, epileptic seizures, and paralysis. The disease is found in developing countries, where porcine cysticercosis is prevalent and undercooked pork is habitually consumed. This study aimed to develop immunochromatography-based test (ICT) kits, using low-molecular-weight antigens purified from cyst fluids of Latin American and Asian genotypes of T. solium . To evaluate the kits, we used 164 serum samples, including 24 from proven/confirmed cysticercosis cases, 110 from cases with other parasitoses, and 30 from healthy individuals. Diagnostic performances were calculated. The sensitivity, specificity, and accuracy were 83.3% (95% CI [62.6–95.3]), 93.6% (95% CI [88.1–97.0]), and 92.1% (95% CI [86.8–95.7]), respectively for the American genotype-based ICT kit, while for the Asian genotype-based ICT kit, they were 87.5% (95% CI [67.6–97.3]), 98.6% (95% CI [94.9–99.8]), and 97.0% (95% CI [93.0–99.0]), respectively. The sensitivity and specificity did not significantly differ between the two ICT kits (exact McNemar’s test; p > 0.05), with a concordance of 93.9%, represented by a Cohen’s kappa of 0.77 ( p < 0.001), indicating substantial agreement. These results indicate that affinity-purified antigens from different geographical isolates can be used for the diagnosis of human cysticercosis. The diagnostic specificities were better than for a previously reported ICT kit that used crude antigen.

Tabanus rubidus (Diptera: Tabanidae) is a significant species of the Tabanidae family, known for its veterinary and medical relevance, and is found across Thailand. This study examines the species distribution, genetic diversity, demographic history, and vector‐pathogen transmission of T . rubidus collected from all six different regions in Thailand. Molecular techniques, utilizing the cox 1 gene, were used to assess genetic diversity, genetic structure, and demographic history. The 16S ribosomal RNA gene was used to detect Anaplasma and Ehrlichia , while the 18S ribosomal RNA gene was used to identify Trypanosoma and Theileria . Tabanus rubidus was found in 23 out of 26 provinces in Thailand, indicating its widespread presence across the country. Negative binomial regression showed that fly abundance was negatively affected by monthly mean temperature but positively influenced by rainfall and site temperature. There was high haplotype diversity and low nucleotide diversity. The median‐joining network network revealed three lineages: Lineage I, which includes Thailand, Malaysia, and Bangladesh; Lineage II, which is found in India and Bangladesh; and Lineage III, which is found in India. Analysis of the demographic history suggested that population expansion started at the end of the Pleistocene. In this study, T. rubidus tested negative for Ehrlichia and Theileria , with Trypanosoma theileri ( T. theileri ) and Anaplasma marginale being the only detected pathogens. These findings are epidemiologically significant, confirming tabanid‐mediated transmission of T. theileri (indicator of trypanosomiasis) and A. marginale (agent of bovine anaplasmosis), both of which pose health risks and economic losses to livestock.

Background: Tick-borne pathogens threaten livestock health and productivity in Southeast Asia. Despite growing regional interest, epidemiological data from the Lao People’s Democratic Republic (Lao PDR) remain insufficient and underreported. Methods: We collected 227 ticks from 63 cattle across northern, central, and southern Lao PDR. Species were morphologically identified as Rhipicephalus microplus and Rhipicephalus sanguineus sensu lato and confirmed by sequencing of the cytochrome c oxidase subunit I gene. Pathogen screening was conducted via polymerase chain reaction (PCR), with sequence identities verified using the GenBank database. Multivariate analyses assessed regional variation. Results: Detected pathogens included the apicomplexan protozoans Babesia bigemina (7%), Babesia bovis (2.6%), and Theileria sp. (10.6%) as well as the bacteria Anaplasma marginale (18.9%), Anaplasma sp. (2.2%), Ehrlichia sp. (6.6%), Ehrlichia minasensis (0.4%), and Aureimonas altamirensis (1.3%). Anaplasma marginale was the most prevalent. Codetections were common, with multiple ticks harboring two or more pathogens. Some double detection occurred more frequently than expected by chance. Conclusions: This study presents the first comprehensive evidence of diverse tick-borne pathogens circulating in cattle ticks from Lao PDR, revealing high genetic similarity to globally recognized strains alongside distinct region-specific detection patterns. Notably, it also constitutes the first report of A. marginale , E. minasensis , B. bigemina , B. bovis , and Theileria sp. in cattle ticks in the country. These findings underscore the urgent need for integrated tick and pathogen surveillance within a One Health framework, with significant implications for regional disease monitoring, livestock health management, and zoonotic risk mitigation. Graphical

This study characterized the microbiota by sequencing the V3-V4 regions of prokaryotic 16S rRNA to investigate the bacterial diversity of fermented fish-entrail sauce (tai-pla or pung-pla) from five provinces in Thailand. Tai-pla samples made from seven different species of fish, three freshwater and four marine, were purchased. Three subsamples from each were analyzed. The samples had salt concentrations ranging from 3 to 13% and pH values ranging from 4.26 to 6.19. The top 35 genera of bacterial taxa by relative abundance were considered in more detail. Lactic acid bacteria (LAB), primarily in the order Lactobacillales (Levilactobacillus, Companilactobacillus, Lactococcus, Latilactobacillus, Weissella, Pediococcus, and Ligilactobacillus), were abundant in several groups of samples, as were halophilic bacteria, including Halanaerobium, Chromohalobacter, and Virgibacillus. Other beneficial bacterial species were frequently detected, including Tetragenococcus halophilus and Tetragenococcus muriaticus. Principal Coordinate Analysis visualization of beta diversity showed distinct bacterial community structures across tai-pla samples prepared with different fish species. Differences between samples may be due to the use of different raw materials, salt concentrations, recipes, processes and fermentation periods. This study provides baseline information on microbial communities and diversity in tai-pla, offering better insights into the production outcomes of traditional products. Further optimization of the fermentation process, such as using beneficial bacterial taxa in starter cultures, may enhance the system of food fermentation, food quality, and flavor control, supporting regulation useful for industrial applications.

Tick-borne pathogens, particularly Babesia and Theileria species, are major threats to cattle production, causing economically significant diseases such as babesiosis and theileriosis. In this study, a real-time SYBR Green PCR assay was developed to detect Babesia and Theileria species in hard ticks (N = 65) and cattle blood samples (N = 143) from Thailand. Using primers targeting the mitochondrial cytochrome b gene for Babesia and the nuclear 18S rRNA gene for Theileria, the assay measured specific melting temperatures (Tm) for each species. The results showed distinct Tm values for Babesia bigemina (74.38 ± 0.04 °C), Babesia bovis (75.7 ± 0.06 °C), Theileria orientalis (74.61 ± 0.03 °C), Theileria sinensis (75.84 ± 0.03 °C), and Theileria annulata (74.06 ± 0.03 °C). The assay demonstrated high specificity, with a cutoff cycle threshold of < 35 cycles and a minimum detectable concentration of 10 copies/μL. Significant species differences in melting curves were confirmed using Tukey’s HSD test (p < 0.05). Theileria orientalis was detected in 8.4% of cattle blood samples, while T. sinensis was found in 25.9%, and B. bigemina in 0.7%. Theileria orientalis was also detected in 7.7% of tick samples, T. sinensis in 16.9%, and B. bigemina in 6.1%. The assay returned negative results for all non-target blood and tissue pathogens tested for specificity. This robust, high-throughput assay is highly effective for monitoring Babesia and Theileria infections, facilitating close surveillance and intervention efforts against tick-borne diseases in cattle.

: Fascioliasis, which is caused by Fasciola hepatica and Fasciola gigantica (F. gigantica), is a public health problem worldwide. Stages infective to humans occur on aquatic plants and in water. The gold standard for diagnosing human infection depends on finding fluke eggs in stool samples. However, this method has low diagnostic sensitivity; eggs from other trematode species might be misidentified as those of Fasciola, and eggs are not voided in ectopic infections. Therefore, serological analysis can support the diagnosis of human fascioliasis. We evaluated whether a new fascioliasis immunochromatographic test kit, which detects specific IgG antibodies using F. gigantica excretory–secretory antigen, can be used for rapid diagnosis from whole blood. The kit (“the fascioliasis whole-blood test kit”) was evaluated under laboratory conditions using 250 whole-blood samples (WBSs), of which 41 were from fascioliasis cases. Results based on simulated WBSs and the corresponding serum samples were compared and showed almost perfect agreement (percent agreement = 97.2%; kappa value = 0.9145). The diagnostic sensitivity and specificity of the test using simulated WBSs were 97.6% and 96.2%, respectively. Comparable values when serum samples were tested were 100.0% and 93.3%, respectively. This kit represents a significant advance because it does not require extensive training of personnel, is easy to use, and can support diagnosis at the bedside or in local and remote hospitals with limited facilities. The kit may also contribute significantly to epidemiological surveys.

: Human strongyloidiasis is an important intestinal parasitic disease that affects public health globally and is transmitted through contact with infective larvae on contaminated soil. Immunocompromised hosts can experience hyperinfection, which can lead to fatal systemic strongyloidiasis. Here, an innovative point-of-care (POC) test kit, the strongyloidiasis immunochromatographic blood test (the NIE-SsIR whole-blood ICT) kit is described. The kit was used to detect anti-Strongyloides IgG antibody in whole-blood samples (WBSs) instead of serum to diagnose strongyloidiasis. The kit is based on a mixture of two recombinant Strongyloides stercoralis protein antigens (NIE and SsIR) and colloidal-gold-labeled conjugates of anti-human IgG antibody to evaluate diagnostic values with simulated and fresh anticoagulated WBSs. The NIE-SsIR whole-blood ICT kit showed potentially high diagnostic values with simulated WBSs, obtained by spiking patients’ sera with red blood cells. The sensitivity, specificity, and positive and negative predictive values were 93.0%, 93.7%, 88.6%, and 96.2%, respectively, at the prevalence of disease simulated under the laboratory conditions of 34.5%. In addition, 18 of 20 fresh anticoagulated WBSs from strongyloidiasis cases were positive, and all 15 WBSs from healthy volunteers were negative. The NIE-SsIR whole-blood ICT kit is a simple and convenient POC testing tool and can possibly be used with fingerstick blood samples, thereby not requiring the drawing of venous blood and separation of the serum. The NIE-SsIR whole-blood ICT kit can assist clinical diagnosis in remote areas and field settings without sophisticated equipment.