Distribution and Abundance of Opisthorchis viverrini Metacercariae in Cyprinid Fish in Northeastern Thailand

To increase public health awareness for prevention of opisthorchiasis caused by eating raw freshwater fish, the distribution and abundance of Opisthorchis viverrini metacercariae (OV MC) was investigated in freshwater fish obtained from 20 provinces in northeastern Thailand between April 2011 and February 2012. A cross-sectional survey was conducted on 12,890 fish consisting of 13 species randomly caught from 26 rivers, 10 dams, and 38 ponds/lakes. Fish, were collected in each of the rainy and winter seasons from each province. Fish were identified, counted, weighed, and digested using pepsin-HCl. Samples were examined for OV MC by a sedimentation method, and metacercariae were identified under a stereomicroscope. OV MC were found in 6 species of fish; i.e., Cyclocheilichthys armatus, Puntius orphoides, Hampala dispar, Henicorhynchus siamensis, Osteochilus hasselti, and Puntioplites proctozysron from localities in 13 provinces. Among the sites where OV MC-infected fish were found, 70.0% were dams, 23.7% were ponds/lakes, and 7.7% were rivers. The mean intensity of OV MC ranged from 0.01 to 6.5 cysts per fish (or 1.3-287.5 cysts per kg of fish). A high mean intensity of OV MC per fish (>3 cysts) was found in 5 provinces: Amnat Charoen (6.5 cysts), Nakhon Phanom (4.3), Mukdahan (4.1), Khon Kaen, (3.5) and Si Sa Ket (3.4). In conclusion, OV MC are prevalent in natural cyprinid fish, with the infection rate varying according to fish species and habitats.

High Prevalence of Opisthorchis viverrini Infection in a Riparian Population in Takeo Province, Cambodia

Opisthorchis viverrini infection was found to be highly prevalent in 3 riverside villages (Ang Svay Chek A, B, and C) of the Prey Kabas District, Takeo Province. This area is located in the southern part of Cambodia, where the recovery of adult O. viverrini worms was recently reported. From May 2006 until May 2010, fecal examinations were performed on a total of 1,799 villagers using the Kato-Katz thick smear technique. In the 3 villages, the overall positive rate for helminth eggs ranged from 51.7 to 59.0% (av. 57.4%), and the percentage positive for O. viverrini was 46.4-50.6% (47.5%). Other helminths detected included hookworms (13.2%), echinostomes (2.9%), Trichuris trichiura (1.3%), Ascaris lumbricoides (0.6%), and Taenia spp. (0.06%). The prevalence of O. viverrini eggs appeared to reflect a lower infection in younger individuals (20 years). Men (50.4%) revealed a significantly higher (P=0.02) prevalence than women (44.3%). The Ang Svay Chek villages of the Prey Kabas District, Takeo Province, Cambodia have been confirmed to be a highly endemic area for human O. viverrini infection.

Ultrasound screening for Opisthorchis viverrini-associated cholangiocarcinomas: experience in an endemic area.

In Thailand, liver cancer is the most common malignancy in males and the third most common among females. In the Northeast region, cholangiocarcinoma (CHCA) is the prevalent type, with Opisthorchis viverrini (OV), an endemic liver fluke, being considered the cause. We evaluated the role of ultrasound (U/S) for cholangiocarcinoma screening as part of a larger cohort to characterize the linkage between liver fluke infestation and CHCA in Khon Kaen (Northeast Thailand). Most people (77%) had normal U/S findings while only 0.5% had suspected CHCA; thus, U/S should be used primarily for those with the highest risk, presenting symptoms and/or being OV positive.

Detection of opportunistic and non-opportunistic intestinal parasites and liver flukes in HIV-positive and HIV-negative subjects.

We assessed the frequency and distribution of infection with opportunistic and non-opportunistic intestinal parasites and the liver fluke, Opisthorchis viverrini, in HIV-seropositive and HIV-seronegative subjects. Age- and sex-matched HIV-seropositive (n = 78) and HIV-seronegative patients (n = 78) from two hospitals in Khon Kaen Province, Thailand, participated in this study from November 1998 to August 2000. These subjects were divided according to the presence of diarrhea and CD4 counts. A single stool sample was obtained and analyzed by using specific techniques. Opisthorchis viverrini, was the most common parasite (19.2%) in each group. The prevalence rates of Cryptosporidium spp (11.5%) and Strongyloides stercoralis (17.9%) in the HIV-seropositive group were significantly (p < 0.05) higher than those in the HIV-seronegative group (1.0% for Cryptosporidium spp and 7.7% for S. stercoralis infections). The prevalences of these two parasites were 28% for Cryptosporidium spp and 20% for S. stercoralis in HIV-seropositives with diarrhea and CD4 counts lower than 100 cells/mm3, and were higher compared with patients without diarrhea or with high CD4 counts. These results suggest that infection with these parasites increases during HIV infection. The epidemiological distribution of Cryptosporidium and S. stercoralis may have implications for AIDS-related diseases.

Epidemiological survey of intestinal parasitic infections of diarrhoeal patients in Nepal and Lao PDR.

Intestinal parasitological survey was conducted to clarify the distribution of intestinal parasites in Nepal and Lao Peoples' Democratic Republic (Lao PDR) from 2001 to 2003. A total of 401 diarrheal fecal specimens were collected from patients aged one to 75 years (250 from Nepal and 151 from Lao PDR). The specimens were examined using the formalin-ether sedimentation (FES) and sucrose centrifugal flotation (SCF) techniques. Nine species (3 Nematoda, 1 Cestoda, and 5 Protozoa) of parasites were recovered from Nepal, whereas seven species (3 Nematoda, 1 Trematoda, and 3 Protozoa) from Lao PDR. Of the parasites detected, (14.4%) was the most common in Nepal, and was Opisthorchis viverrini (29.8%) in Lao PDR. Infection rates were markedly different among age groups in both countries; higher rates were observed in age groups of 10-29 years than in 0-9 years group. Scanning electron microscopic observation revealed that trematoda eggs recovered from Lao PDR were identified as O. viverrini, which could not be done by a light microscope.

An epidemiological study on Opisthorchis viverrini infection in Lao villages.

The prevalence of liver fluke, Opisthorchis viverrini, infection in rural and urban communities was studied in Khammouane Province in Lao PDR. The infection was the commonest among the villagers examined, showing the positive rates of 52.9% and 55.0% in two rural communities and 60.7% in an urban community, respectively. The infection rate reached up to 20% or more within 4 years after birth and increased with age to a plateau in the age group over 20 years. The highest prevalence rate was over 80% in the age group of 35-54 years. The age-related patterns of infection in males and females were almost the same, although there were some sex-related differences by village and by age group. Infection with Opisthorchis viverrini appears to be a serious public health problem strongly associated with the frequent eating habit of raw fish in low land Laos.

A study of liver fluke infection in Sukhothai, Thailand.

A cross sectional survey on opisthorchiasis was conducted from August to September 1933 in Tambon Nai Muang, Sawankhalok district, Sukhothai Province. The 1,964 villagers were stool examined and it was found that 384 of them had liver fluke infection. Of these 1,608, who were older than 12 years, were interviewed in order to detect potential risk factors of liver fluke infection. The study found that the population at risk were those older than 24 years of age (p < 0.05) and the possible risk factor was eating incomplete cooked fresh water fish (p < 0.05). The food prepared from incomplete cooked fish which was harmful to the villagers were pickle fish so called pla ra (p < 0.05) and papaya salad with pickle fish so called som tam pla ra (p < 0.05).

Opisthorchis viverrini metacercaria in Thai freshwater fish.

Examination for metacercaria in freshwater fish, the common intermediate hosts of Opisthorchis viverrini was carried out during 1992-1996. The 4-year survey of fish from markets in 14 provinces revealed that metacercariae of O. viverrini were found in fish from Udon Thani, Sa Kaeo and Prachin Buri Provinces; fish from Aranyaprathet district had the highest positive rates (25-28%). Fish from 12 provinces were found to be positive with heterophyid metacercariae, namely: Haplorchis pumilio, H. taichui, H. yokogawai, Stellantchasmus falcatus, Centrocestus formosanus and Haplorchoides cahirinus. It was also observed that the prevalence of O. viverrini metacercaria in fish decreased markedly during the last 10 years.

Epidemiology of opisthorchiasis and national control program in Thailand.

Opisthorchiasis is a disease caused by opisthorchis trematode commonly known as liver fluke. In Thailand Opisthorchis viverrini is the only parasite of opisthorchiasis, the first case of opisthorchiasis was reported in 1911 by Leiper from the autopsy of corpse in Chiang Mai. Later on Sadun in 1953, Harinasuta and Vajjarasthira in 1961, and Wykoff in 1965 had demonstrated a complete life cycle of O. viverrini. History of opisthorchiasis control has dated back to 1950 as a small scale helminthiosis control program in some high risk areas. Following a number of studies and trial projects of the Faculty of Tropical Medicine of Mahidol University, the national liver fluke control program has been developed and operated under different National Public Health Development Plans. Presently, the program is being operated in some central provinces, and all northeastern and northern provinces of Thailand. The main strategies for liver fluke control are comprised of 3 interrelated approaches, namely stool examination and treatment of positive cases with praziquantel for eliminating human host reservoir; health education for a promotion of cooked fish consumption for preventing infection; and the improvement of hygienic defecation for transmission interruption. Data obtained from liver fluke control operation in 1996 show the countrywide prevalence of opisthorchiasis at 21.5% with a variation in all age groups. Differences of infection are also found among geographical regions. A large percentage of infection is mild infection. Comparing figures of 1981 and 1991, there is a considerable increase in the central and northern regions, while the northeastern region has significant decrease.

The impact of a decade long opisthorchiasis control program in northeastern Thailand.

Based on the figures obtained from a regional survey in 1981, the overall prevalence of opisthorchiasis in northeastern Thailand was as high as 34.6%. Traditional consumption of improperly cooked fish dishes and unhygienic defecating habits among rural inhabitants have been recognized as significant factors determining such high prevalence of this liver fluke in that area. Following experience of Mahidol University's Faculty of Tropical Medicine in Thailand of treating liver fluke infected individuals with praziquantel, the Ministry of Public Health started a liver fluke control program in the northeast as special service units in 1984 in 4 provinces. In 1988, the program was expanded to cover all 19 northeastern provinces, services being rendered at a community level covering stool examination and treatment of positive cases. Intensive health education was integrated as one element of liver fluke control program. During the early phase of operation (attack phase) a mobile stool examination team was the crucial strategy for active service. Following the attack phase, passive service was operated at each health service facility provided that results of population based program evaluation in each area showed a low prevalence according to the set target. Regionwide assessments in 1991 and 1992 indicated a declining trend of prevalence from 34.6% to 30.19% and 24.01%, respectively. In 1994, a population-based sampling survey was carried out to measure the effect of the liver fluke control program over 10 years operation (1984-1994) on both behavioral outcomes and prevalence impact. Sample for the assessment of behavior were 1,268 individuals, while another 1,912 samples were used for determining disease prevalence. Health related behaviors regarding frequent consumption of uncooked fish dishes decreased from 14% in 1990 to 7% in 1994 while occasional consumption remains as high as 42%. The prevalence of opisthorchiasis is 18.5% with a large variation in infection rate (5.20-56.25%).

Application of hazard analysis critical control point (HACCP) as a possible control measure for Opisthorchis viverrini infection in cultured carp (Puntius gonionotus).

Opisthorchiasis due to Opisthorchis viverrini and transmitted through infected freshwater cyprinoid fish (carps) affects more than 8 million people in Thailand, People's Democratic Republic of Lao, and Vietnam. The Hazard Analysis Critical Control Point (HACCP)-concept has been recommended by FAO and WHO to be included in programs to control foodborne trematode infections (FBT). HACCP is a multifactorial approach to control food hazards through surveillance of diseases, foods, and operations and education. This study describes the first attempt to apply HACCP to the prevention and control of Opisthorchis viverrini in pond culture carp (Puntius gonionotus). The experiment was designed and carried out by a multidisciplinary "HACCP team" including experts in the field of public health, parasitology, epidemiology, aquaculture, fisheries extension and fish inspection. The investigation was performed in two fish ponds in the District of Sun Pa Tong, Chiang Mai, Thailand. In the experimental pond, fish was cultured according to HACCP principles and compared with the control pond, which followed conventional aquaculture practices. Water supply to the pond, fish fry, fish feed and pond conditions during the growing period were identified as critical control points (CCPs). Hazards were identified and analyzed, as well as control measures, critical limits, monitoring procedures, corrective actions, and record keeping developed for each one of the above CCPs. Complete pond preparation, particularly aiming to eliminate contamination of pond water with O. viverrini eggs, fish infected with parasite meacercariae and the first intermediate host (Bithynia spp), was conducted. After the pond was filled with water, O. viverrini metacercaria-free fry were released into the pond. The preliminary results obtained indicate that HACCP-based principles applied to carp pond culture could be used as a strategy to prevent and control O. viverrini. Further studies should be undertaken aiming to confirm these preliminary results.

Epidemiological aspects of aquaculture in relation to fish borne trematodiasis in Malaysia.

Epidemiological studies have been conducted to determine the association between fish and disease. The fish were obtained from rivers, streams, ponds and lakes but few from aquaculture farms. While no defined studies have been carried out in Malaysia, baseline data show that fish obtained from aquaculture farms (mixed farming) contributed to cases of opisthorchiasis and clonorchiasis.

Prevalence and intensity of Opisthorchis viverrini in rural community near the Mekong River on the Thai-Laos border in northeast Thailand.

The prevalence and intensity of Opisthorchis viverrini in fourteen villages in Nakhon-Phanom province, Northeast, Thailand have been investigated. Overall prevalence of O. viverrini infection was 66.4 per cent in a total population of 2,412 individuals. The prevalence was 18.5 per cent in children under 5 years, 38.9 per cent in those aged 5-9 years, and ranged from 64.9 per cent to 82.2 per cent in the age group above 10 years. The intensity of O. viverrini infection increased with age. The mean faecal egg output was highest in the 30-34 year age group and remained relatively constant through older ages. In all age groups the prevalence and intensity of infection in both men and women were similar. The population was divided according to the presence and intensity of infection as follow, 33 per cent were uninfected, 59 per cent had light infections (less than 1,000 eggs per g of faeces; EPG), 7 per cent had moderate infections (1,000-10,000 EPG), and 1 per cent had heavy (greater than 10,000 EPG). Other important intestinal infections found in this community are hookworm, Taenia spp. and Trichuris trichiura with the prevalence of 17.9 per cent, 1.1 per cent and 1.1 per cent respectively.

Influence of water current on the distribution of Opisthorchis viverrini infection in northeastern villages of Thailand.

The prevalences of Opisthorchis viverrini infection in the villages on the banks of rivers and those far from the rivers were studied in Loei and Nong Khai Provinces in Thailand. Most of the people examined in the present study were agriculturalists. The overall prevalence of O. viverrini infection was 41.3%. The prevalence of infection in males and in females in the villages far from the rivers were 52.6% and 51.7%, respectively, while the percent of people in the villages on the banks with infection were 27.9% and 21.7%, respectively. Prevalence of infection among the people residing far from the rivers was higher than those residing on the banks. This was observed despite the higher recording of raw fish consumption in villages on the banks. Infection level increased sharply in the age-group 6-10 years old among people residing far from the rivers. High prevalences of infection were observed in age groups from 11 to 50 years. Both types of villages had a similar proportion of houses with sanitary latrines.