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Opisthorchis viverrini and minute intestinal fluke (MIF) infections are heavily epidemic in northeastern Thailand. Their primary cause is eating raw or undercooked cyprinid fishes, and they cause health problems in the human digestive system. In cases of liver fluke, these parasites can go through the bile duct system, which may cause cholangiocarcinoma (bile duct cancer). When a medical doctor suspects that a patient is infected with parasites, they typically request a stool analysis to determine the type of egg parasites using microscopy. Both parasites have similar characteristics, thus, it is necessary for a specialist to identify the specific type of egg parasites present. Many automatic systems have been developed using deep learning to assist doctors in diagnosing the type of egg parasite. In this study, we proposed three models of deep learning architectures and created voting ensembles to analyze egg parasite images. Images of similar liver fluke eggs and MIF eggs were taken from the Parasitology Laboratory, Faculty of Medicine, Khon Kaen University, Khon Kaen, Thailand. Image data augmentation is used to expand images from different perspectives and assist the system in acquiring a greater variety of images. Three models performed effectively, by employing the hard voting ensemble, the accuracy increased to 86.67%, while for the second group, the accuracies reached 68.00%, 76.00%, and 77.33%, respectively. Using the soft voting ensemble, the accuracy improved to 79.33%. These outcomes highlight the potential of ensemble deep learning in image classification. Furthermore, these results align closely with those achieved by several experts in image classification. Hence, a promising ensemble approach can aid doctors in accurately classifying images of egg parasites. [ABSTRACT FROM AUTHOR]

Global liquid chromatography mass spectrometry (LC-MS) profiling in a Thai population has previously identified a urinary metabolic signature in Opisthorchis viverrini -induced cholangiocarcinoma (CCA), primarily characterised by disturbance in acylcarnitine, bile acid, steroid, and purine metabolism. However, the detection of thousands of analytes by LC-MS in a biological sample in a single experiment potentially introduces false discovery errors. To verify these observed metabolic perturbations, a second validation dataset from the same population was profiled in a similar fashion. Reverse-phase ultra-performance liquid-chromatography mass spectrometry was utilised to acquire the global spectral profile of 98 spot urine samples (from 46 healthy volunteers and 52 CCA patients) recruited from Khon Kaen, northeast Thailand (the highest incidence of CCA globally). Metabolites were differentially expressed in the urinary profiles from CCA patients. High urinary elimination of bile acids was affected by the presence of obstructive jaundice. The urine metabolome associated with non-jaundiced CCA patients showed a distinctive pattern, similar but not identical to published studies. A panel of 10 metabolites achieved a diagnostic accuracy of 93.4% and area under the curve value of 98.8% (CI = 96.3%–100%) for the presence of CCA. Global characterisation of the CCA urinary metabolome identified several metabolites of biological interest in this validation study. Analyses of the diagnostic utility of the discriminant metabolites showed excellent diagnostic potential. Further larger scale studies are required to confirm these findings internationally, particularly in comparison to sporadic CCA, not associated with liver fluke infestation. [ABSTRACT FROM AUTHOR]

Despite significant advances in understanding the role of the immune response in Opisthorchis viverrini-associated carcinogenesis, little is known about how infection induces gall bladder disease. This study investigated whether mast cells are activated in cholecystitis associated with O. viverrini, gall bladder specimens from ninety-two patients who had undergone cholecystectomy at the Khon Kaen Regional Hospital, Khon Kaen, Thailand. Two representative sections from the body of fresh gall bladder tissue were fixed in Carnoy's solution and embedded in paraffin wax. The paraffin sections were stained for mast cells and IgE plasma cells by the double histochemical and immunohistochemical method. The cells in the epithelium, lamina propria, muscular layer, and subserosa were counted and expressed as cells per square millimeter. The gall bladder bile was examined for the presence of O. viverrini eggs. Significantly higher mean mast cell numbers were found in the lamina propria (221.41 ± 16.01 vs 116.97 ± 14.61 cells per mm2; P < 0.005) of egg positive compared to egg negative groups, respectively. No comparable differences in mast cell number were observed in other layers. IgE plasma cells were rarely seen. The results suggest that mast cell hyperplasia occurs during cholecystitis in association with opisthorchiasis and may play a role in the pathogenesis of the disease. [ABSTRACT FROM AUTHOR]

• The infection rate of Ov does not dramatically decline (34.78%). • No significant difference in the GPS range distance in Ov infected cats. • Compositional analysis suggested that the domestic cats preferred urban habitats. • PE results suggest the villagers' awareness is not enough for the effective control. In the endemic area of the liver fluke (Opisthorchis viverrini) infection, effective control of opisthorchiasis using One Health approaches requires transmission interruption in all hosts in the parasite life cycle. Owned domestic cats (Felis catus) are considered an important species among mammalian reservoirs. Similar to humans, household cats become infected by eating raw, contaminated fish and spread the O. viverrini eggs to the environment by defecation. In rural areas, cat owners usually feed their cats with human food or food waste, especially fish dishes. There is a high risk for cats to become infected and deposit eggs in the peri-household environment. The lack of research on domestic cats led us to study their roaming behavior, roaming range, habitat preference and risk of disease transmission. GPS loggers were harnessed to 28 domestic cats in 7 villages around Lawa Lake, the endemic area of human opisthorchiasis. All cats were examined for the O. viverrini infection status using modified formalin-ether concentration technique (MFECT) before application of the GPS device used to observe and record their roaming distance and range area. Then, composition analysis for habitat selection was performed using the location data. Additionally, cat owners were interviewed using participatory epidemiologic methods for their perceptions and knowledge about the reservoir hosts of this trematode. Our study suggested that the prevalence of O. viverrini in cats in this area has not declined appreciably in comparison with our previous study four years ago, although the treatment and control has been applied intensively in humans. The longest distance that individual cats roamed ranged from 51 to 363 meters (mean±SD = 154.48±90.71). The cats traveled tracks around their home. There was no significant difference in the average distance that cats ranged between O. viverrini infected group (154.13±72.80 meters) and the non-infected group (154.67±101.39 meters) was found. Compositional analysis suggested that the domestic cats preferred urban habitats where food and safety were probably more available. It appears that the main transmission route for infection of cats was contaminated fish fed to cats or fish that remained in their houses. Therefore, owners should refrain from feeding their cats potentially contaminated foods, food scraps and raw fish remains and prevent their cats from accessing them. Moreover, the participatory epidemiology results suggest that awareness of villagers is not enough for the effective and sustainable control of the disease. [Display omitted] [ABSTRACT FROM AUTHOR]

Opisthorchiasis caused by Opisthorchis viverrini is an important foodborne trematodiasis in Thailand, Laos and Cambodia. Interestingly, the opisthorchiasis endemic region overlaps with an area of leptospirosis emergence. Here we report an association between opisthorchiasis and leptospirosis in Thailand. Of 280 sera collected from villagers living around the Lawa wetland complex in Khon Kaen province, 199 (71%) were seropositive for leptospirosis by immunochromatography. Individuals with O. viverrini infection had a significantly higher rate of leptospirosis than those without ( P = 0.001). Significant higher leptospirosis prevalence was found in males than females ( P = 0.002). However, females but not males with O. viverrini infection showed a significantly higher seroprevalence of leptospirosis. Twenty-one of 35 environmental samples from the lake (water, mud and fish skin mucus) were positive for Leptospira spp. DNA sequencing, sequence alignment, and phylogenetic analysis of some positive nested PCR products revealed both pathogenic and intermediate pathogenic strains of Leptospira in the samples. Strikingly, O. viverrini metacercariae from the fish were positive for L. interrogans. These results suggest a close association between opisthorchiasis and leptospirosis. Contact with water, mud or eating raw fish harboring liver fluke metacercariae may be risk factors for Leptospira infection. [ABSTRACT FROM AUTHOR]

Khon Kaen Province in northeast Thailand is known as a hot spot for opisthorchiasis in Southeast Asia. Preliminary allozyme and mitochondrial DNA haplotype data from within one endemic district in this Province (Ban Phai), indicated substantial genetic variability within Opisthorchis viverrini. Here, we used microsatellite DNA analyses to examine the genetic diversity and population structure of O. viverrini from four geographically close localities in Khon Kaen Province. Genotyping based on 12 microsatellite loci yielded a mean number of alleles per locus that ranged from 2.83 to 3.7 with an expected heterozygosity in Hardy–Weinberg equilibrium of 0.44–0.56. Assessment of population structure by pairwise FST analysis showed inter-population differentiation (P<0.05) which indicates population substructuring between these localities. Unique alleles were found in three of four localities with the highest number observed per locality being three. Our results highlight the existence of genetic diversity and population substructuring in O. viverrini over a small spatial scale which is similar to that found at a larger scale. This provides the basis for the investigation of the role of parasite genetic diversity and differentiation in transmission dynamics and control of O. viverrini. [ABSTRACT FROM AUTHOR]

Abstract: Khon Kaen, a northeastern province of Thailand, has been considered as one of the human opisthorchiasis endemic areas with continuing high prevalence. Unsuccessful eradication of the disease is probably from the culture of eating raw and undercooked fish of local residence and the parasitic persistency in animal reservoir hosts, such as cats and dogs. In cooperation with the other human opisthorchiasis control programs in an endemic area of 29 villages in Ban Haet, Ban Phai, Chonnabot and Muncha Khiri Districts, Khon Kaen, this study investigated the prevalence of Opisthorchis viverrini infection using a formalin-ether sedimentation method as the gold standard, and hematology and blood chemistry of the reservoir hosts in this endemic area. The results showed that cats had much higher prevalence (76 of 214, 35.51%) than dogs (3 of 821, 0.37%). Hematology between the infected and uninfected cats was not different. Complete blood count and biochemistry reflected some altered hepatic functions. However, only severely infected cats showed apparent clinical signs, including lethargy, diarrhea, ocular and nasal discharges. Moreover, the ultrasonogram of infected cats with very high egg per gram (>1500 EPG) showed apparent thickening of the gall bladder wall with hyperechoicity of hepatic parenchyma. This study suggests that cat is the most important animal reservoir of human opisthorchiasis, especially in this endemic area. It is also interesting that villages with infection are mostly located in the vicinity of Chi River and two large water reservoirs (Lawa and Nong Kongkaew Lakes), but people without infection were away from Chi River, on the south of Kudkhow Lake. Further investigation on this particular geofactor is essential for effective opisthorchiasis control programs. [Copyright &y& Elsevier]

The population genetics of 317 individual Opisthorchis viverrini from Khon Kaen Province Thailand, from 4 different years and 4 cyprinid fish species was examined using multilocus enzyme electrophoresis of enolase (Enol), phosphoglucomutase (Pgm) and triose phosphate isomerase (Tpi). Allele and genotype frequencies for Enol and Pgm were consistent irrespective of year or host species. No heterozygote deficiency was detected for Enol. Significant heterozygote deficiencies were detected in 3 of 4 years for Pgm. For Tpi, allele frequencies of the most common allele and genotype frequency varied between years and among individuals from different host species. Heterozygote deficiencies for Tpi were detected in 2 years. No significant heterozygous deficiencies were detected among O. virerrini from different fish species in 2005, except at Pgm and Tpi from Puntioplites protozsron. There was no statistical significance in pairwise FST values between O. viverrini from Cyclocheilichthys armatus in different years or different host species in 2005. Significant departures from Hardy-Weinberg expectations and a high rate of gene flow in a population of O. viverrini are discussed in terms of self- and cross-fertilisation, natural selection, non-random mating, the Wahlund effect, presence of null alleles, intensity of infection, biology and ecology of their intermediate cyprinid hosts. [ABSTRACT FROM AUTHOR]

Abstract: A polymerase chain reaction (PCR) assay was evaluated for detection of Opisthorchis viverrini eggs in the stool specimens of light and heavily infected individuals in Khon Kaen province of Thailand. A total of 75 fecal specimens were analyzed by PCR following DNA extraction. All the microscopically positive samples were positive by PCR, while 23 of 30 (76.6%) microscopically negative samples were also PCR positive. The sensitivity of the assay was 5 eggs/g of stool. This method is potentially useful in the diagnosis of human opisthorchiasis in endemic areas for treatment and in epidemiological investigations. [Copyright &y& Elsevier]

• Praziquantel treatment for opisthorchiasis alleviates parasite burden • 30.8% of participants showed no relapse of hepatobiliary abnormality over five years • 37.5% of participants showed persistent hepatobiliary abnormality Infection of the liver fluke, Opisthorchis viverrini (OV) is an important public health problem in northeast Thailand and adjacent countries, where people have a habit of eating raw or undercooked fish. A community case-control study was carried out with 8,936 participants from 89 villages, in Khon Kaen province, Thailand. There were 3,359 OV-infected participants all of whom underwent ultrasonography of upper abdomen for the evaluation of hepatobiliary morbidity. The participants with advanced periductal fibrosis (APF) by ultrasound (n = 785) were invited to undergo annual follow-up ultrasonography for five years after praziquantel treatment. The sonographer was blinded with respect to status of OV infection at each visit. The study findings revealed variability in the study population profile of the hepatobiliary morbidities before and after praziquantel treatment over the follow up interval. At the end of the study, 32 (30.8%) out of 104 participants showed no relapse of APF whereas, by contrast, 39 (37.5%) participants showed relapse or persistent APF since the outset of the study (≥ two consecutive visits). The APF in most follow-up visits was significantly associated with male sex, with intrahepatic duct stones, with the width of the gallbladder "pre" minus "post" fatty meal, and with the ratio of left lobe of the liver to aorta. Five cases of suspected cholangiocarcinoma were observed over the five years of follow-up. This long-term ultrasound follow-up study demonstrates a significant incidence of persistent APF in over one-third of opisthorchiasis cases after praziquantel treatment, findings that support the prospect of ongoing cholangiocarcinogenesis long after successful elimination of liver fluke infection among the population. [Display omitted] [ABSTRACT FROM AUTHOR]