Effects of Environmental Factors and Infecting Trematodes on the Size and Inorganic Elements of Bithynia siamensis goniomphalos Snails in Northeast Thailand

Infection with the foodborne trematode, Opisthorchis viverrini, is a major public health issue in southeast Asia. The freshwater snail, Bithynia siamensis goniomphalos, is an intermediate host of O. viverrini and other trematode species. Understanding the effects of environmental conditions and infecting trematodes on B.s. goniomphalos snails is thus crucial for the potential influences on trematode transmission. This study measured environmental variables of water and soil properties, and analyzed B.s. goniomphalos snails for their trematode infection, snail shell length, and inorganic elemental concentration, from 30 localities in northeast Thailand. The results showed that prevalence of trematode infection in B.s. goniomphalos was 3.82%. Nine types of trematode cercariae were identified, with virgulate type 1 as the most common (1.23%). Opisthorchis viverrini-infected snails were mostly found in low-humic gley soils in Sakon Nakhon Province, and were associated with water dissolved oxygen and soil pH. Compared with uninfected snails, larger sizes were observed in virgulate type 1 and pleurolophocerca-infected snails, whereas hypercalcification was noticed in virgulate type 1, virgulate type 3, and pleurolophocerca-infected snails. Infected snails were more sensitive toward environmental conditions, possibly because of the dynamic parasitic processes between trematodes and hosts. Among the environmental factors, soil texture (i.e., sand, silt, and clay compositions) exhibited more significant correlations with B.s. goniomphalos shell characteristics regardless of the trematode infection types. The findings of this study underscore the need to consider the effects of environmental conditions and trematode species-specific pathogenic processes for a more effective and sustainable parasitic control and prevention effort.

Efficiency of the Stool-PCR Test Targeting NADH Dehydrogenase (Nad) Subunits for Detection of Opisthorchis viverrini Eggs

Opisthorchis viverrini infection is the major parasitic infection problem in Southeast Asian countries, and long-term infection will lead to cholangiocarcinoma (CCA), the bile duct cancer. The early diagnosis of O. viverrini infection may interrupt the progression of the opisthorchiasis and other related illnesses, especially CCA. The current diagnostic procedure is stool examination by microscope-based methods such as direct smear and concentration techniques but it is limited by low parasite egg numbers. The molecular diagnosis prompts the chance to evaluate the light infection with low number of parasite eggs but is currently inconvenient for routine use due to special equipment requirement and unstable sensitivities. Our present study aims to establish the efficiency of OvNad subunits, the mitochondrial gene, for introducing as a potential diagnostic target by conventional PCR, the cheapest and easiest molecular procedure. A total of 166 stool samples were investigated microscopically by the PBS-ethyl acetate concentration technique (PECT); 75 samples were O. viverrini positive with 28 samples that were positive with single parasite (hookworm, A. lumbricoides, S. stercoralis, Taenia spp., and T. trichiura), 11 samples were with mixed infection, and 52 samples were without parasite detection. The detection limits of OvNad subunits were evaluated in artificially spiked samples containing 0, 1, 5, 10, 20, 50, and 100 Ov-eggs. The result suggested that the best detection efficacy was of OvNad5 that had exact detection limits at only 5 eggs. In the PCR amplification of OvNad subunits, there exist 100% specificities with varied sensitivities from 64%, 88%, 80%, and 100% of OvNad1, OvNad2, OvNad4, and OvNad5, respectively. OvNad subunits were amplified specifically without cross reactivity with the other collected parasites. Our study established that OvNad subunits, especially OvNad5, are the potent candidates for PCR amplification of stool containing Ov-eggs with high confidential sensitivity, specificity, PPV, and NPV even in the light infection that would be a benefit for developing as a routine diagnosis of O. viverrini infection.

Association of comorbidity between Opisthorchis viverrini infection and diabetes mellitus in the development of cholangiocarcinoma among a high-risk population, northeastern Thailand

Background Cholangiocarcinoma (CCA) is a category of lethal hepatobiliary malignancies. Previous studies have found that Opisthorchis viverrini infection and diabetes mellitus (DM) are closely correlated with CCA. However, few studies have discussed the association of CCA with a combination of both O. viverrini infection and DM. This study aimed to assess the correlation of CCA with various combinations of O. viverrini infection and DM among a high-risk population in northeastern Thailand. Methodology This study included participants from 20 provinces in northeastern Thailand who had been screened for CCA in the Cholangiocarcinoma Screening and Care Program (CASCAP) between 2013 and 2019. Histories of O. viverrini infection and DM diagnosis were obtained using a health questionnaire. CCA screening used ultrasonography with a definitive diagnosis based on histopathology. Multilevel mixed-effects logistic regression was performed to quantify the association, which is presented as adjusted odds ratios (aOR) and their 95% confidence intervals (CI). Principal findings Overall, 263,776 participants were included, of whom 32.4% were infected with O. viverrini, 8.2% were diagnosed with DM, and 2.9% had a history of both O. viverrini infection and DM. The overall rate of CCA was 0.36%. Of those infected with O. viverrini, 0.47% had CCA; among those with DM, 0.59% had CCA and among those infected with O. viverrini and had DM, 0.73% had CCA. Compared with participants who were not infected with O. viverrini and were non-DM, the aOR for those infected with O. viverrini and with DM was 2.36 (95% CI: 1.74–3.21; p-value <0.001). Conclusions The combination of O. viverrini infection and DM was highly associated with CCA, and these two conditions had a combined effect on this association that was greater than that of either alone. These findings suggest that CCA screening should have a strong focus on people with a combination of O. viverrini infection and DM.

Prevalence of Opisthorchis viverrini and its Associated Risk Fac-tors in the Phon Sawan District of Nakhon Phanom Province, Thailand

Background: Opisthorchis viverrini is one of the endemic helminths in Thailand. Nonetheless, O. viverrini still exist and raise concerns regarding public health. This study aimed to evaluate the current prevalence of O. viverrini infections and underlying risk factors among rural communities in Northeast Thailand. Methods: A cross-sectional study was conducted between Sep and Dec 2019 in Phon Sawan district in Nakhon Phanom Province, Thailand. The participants were selected using a voluntary sampling method after the proportional allocation of the total sample size. Demographic data were collected using a standardized questionnaire. One stool sample was collected from each participant and the presence of O. viverrini and other intestinal parasite infections were determined using the modified Kato-Katz technique. Results: Overall, 250 participants aged 21 yr or older were enrolled. Overall, the prevalence of O. viverrini and Strongyloides stercoralis was 24% (95% confidence interval [CI]: 18.8-29.8) and 1.2% (95% CI: 0.2-3.5), respectively. Of the positive O. viverrini cases, the parasite rate significantly differed between gender, age group and in those who had a history of eating raw fish and taking the anti-helminthic drug (all P<0.05). Nevertheless, multivariate regression analysis among O. viverrini cases revealed that only being male (adjusted odds ratio [aOR] 1.9 [95% CI: 1.1 – 3.6], P = 0.033) and aged ≥61 yr (aOR 6.7 [95% CI: 1.4 – 32.5], P=0.018) were positively associated with a higher risk of O. viverrini infection. Conclusion: Opisthorchiasis and strongyloidiasis are still endemic in this area and there is a need for projects to eliminate these parasites.