Fishborne Zoonotic Trematode Metacercariae Research Articles

Prevalence of fish-borne zoonotic trematode infection in Jilin Province, China.

Fish-borne zoonotic trematodes (FZTs) are the most serious food-borne parasites in Asia and have become a burden to public health and a new challenge in food safety. In Jilin Province, China, the prevalence of FZTs in intermediate and definitive hosts has not been extensively explored. In the present study, we investigated the prevalence of FZTs in Jilin Province, China. From July to November 2020, a total of 132 freshwater snails (the first intermediate host of FZTs), 4122 wild freshwater fishes (the second intermediate host of FZTs) and 143 fecal samples from canines, ducks and swine (the definitive host of FZTs) were collected from the Yitong River basin of Jilin Province. FZT species were identified by morphological observation combined with internal transcribed spacer (ITS) sequence analysis. The prevalence of FZTs was then calculated. The results showed that the prevailing species of FZTs in Jilin Province, China, were Clonorchis sinensis, Metorchis orientalis and Echinochasmus japonicus. The total prevalence of FZTs was 29.74% (1226/4122) in fish, the total infection rates were 2.27% (3/132) in snails, 75.00% (21/28) in canines and 37.18% (29/78) in ducks. The coinfection rates of the two trematodes were 13.39% (552/4122) in fish, 35.71% (10/28) in canines and 7.69% (6/78) in ducks. The coinfection rate of the three flukes was 2.60% (107/4122) in fish. Nine of the 12 fish species examined were infected with FZT metacercariae.

Infection Status of Zoonotic Trematode Metacercariae in Fishes from Vientiane Municipality and Champasak Province in Lao PDR.

The infection status of fishborne zoonotic trematode (FZT) metacercariae was investigated in fishes from 2 localities of Lao PDR. Total 157 freshwater fishes (17 species) were collected in local markets of Vientiane Municipality and Champasak Province in December 2010 and July 2011, and each fish was examined by the artificial digestion method. Total 6 species of FZT metacercariae, i.e., Opisthorchis viverrini, Haplorchis taichui, Haplorchis yokogawai, Haplorchis pumilio, Centrocestus formosanus, and Procerovum varium, were detected in fishes from Vientiane Municipality. The metacercariae of O. viverrini were detected in 50 (49.5%) out of 101 fishes (6 species), and their average number was 154 per fish infected. The remaining 5 species of heterophyid metacercariae were detected in 36.8%, 65.8%, 9.4%, 23.9%, and 5.1% fishes examined, and their average densities were 12, 1,038, 4, 15, and 13 per infected fish, respectively. In fishes from Champasak Province, 3 species of FZT metacercariae, i.e., O. viverrini, H. taichui, and H. yokogawai, were detected. Only 2 O. viverrini metacercariae were found in only 1 Barbonymus schwanefeldi. The metacercariae of H. taichui and H. yokogawai were detected in 60.0% and 50.0% of fishes examined, and their average densities were 47 and 28 per fish infected. By the present study, it has been confirmed that several species of FZT metacercariae are prevalent in fishes from Vientiane Municipality, with P. varium being a new member of FZT in Lao PDR. In comparison, FZT metacercariae are less prevalent in fishes from Champasak Province.

Effect of pond water depth on snail populations and fish-borne zoonotic trematode transmission in juvenile giant gourami (Osphronemus goramy) aquaculture nurseries

Infection with fish-borne zoonotic trematodes (FZT) is an important public health problem in many parts of Southeast Asia. People become infected with FZT when eating raw or undercooked fish that contain the infective stage (metacercariae) of FZT. The parasites require specific freshwater snails as first intermediate host and a variety of fish species, both wild caught and cultured, as second intermediate host. Aquaculture production has grown almost exponentially in SE Asia and in order to produce fish free from FZT metacercariae, it is important to mitigate factors promoting transmission to fish. Here we report results from a cross-sectional study to look at the association between pond depth and infection with FZT in giant gourami nursery ponds. Density of intermediate host snails was positively associated with pond depth (count ratio associated with a 1m increase in pond depth was 10.4 (95% C.L.: 1.61–67.1, p<0.5)) and this may partly explain the higher prevalence and intensity of FZT infection in juvenile fish. High fry stocking density (>200frym−3) was associated with lower host snail density (count ratio=0.15) than low stocking density (<100frym3). Ponds stocked with 100–200frym−3 had snail counts 0.76 (95% C.L.: 0.33–1.75, p n.s.) of those in ponds stocked with fry density of <100frym−3. Since density of intermediate snail hosts was associated with FZT transmission to fish, effort should be taken to reduce snail density prior to stocking the fry, but focus should also be on habitats surrounding ponds as transmission may occur through cercariae produced outside ponds and carried into ponds with water pumped into ponds.

The effectiveness of different intervention strategies for the prevention of zoonotic metacercariae infection in cultured fish

Fish-borne zoonotic trematodes (FZT) are a major public health problem. It is estimated that in Vietnam 26,366 “disability-adjusted life years” (DALYs) are lost due to FZT. Fish from aquaculture are a main source of protein and of great economic importance in both rural and urban areas. The objective of this study was to compare the impact on transmission of FZT to juvenile fish of drug treatment intervention of final hosts, and pond management interventions on fingerlings produced in nurseries producing Rohu carp in Northern Vietnam. The study design was a parallel group design of nurseries with two intervention groups; a drug treatment of human and animal groups and a farm management group (control of snail vectors and fecal pollution of pond). A third group (non-intervention) served as control. Fish were examined for FZT metacercariae prevalence and intensity before and after interventions, along with density of potential host pond snails. Fecal exams for FZT infections in farm household humans, dogs and cats were also performed. The results showed that although FZT prevalence in fish was not significantly reduced, the intensity of metacercariae in fish from ponds receiving management interventions was reduced 91.7% compared to before interventions. The intensity of FZT was also significantly lower in the pond management group, compared to the drug treatment group and the control group after interventions. The results demonstrate that improving farm and pond management practices can be effective in reducing FZT infections without using drug treatment. This study also highlighted certain obstacles to attaining higher levels of protection for which more research is needed. A major challenge is the ability to completely isolate (or bio-secure) the pond from its surrounding environment, especially surrounding water sources which may harbor snails and cercariae that can contaminate the pond through water resupply or runoff. Another is the unknown risk from piscivorous birds, host for intestinal FZT. The strategies employed in this study now need long-term evaluation on a regional basis, the outcome of which could be the development of good management practices crucial to ensuring a good quality product that benefits farmers and the consumers.

A Cross-Sectional Study on the Potential Transmission of the Carcinogenic Liver Fluke Opisthorchis viverrini and Other Fishborne Zoonotic Trematodes by Aquaculture Fish

Throughout Southeast Asia and China, eating raw and or partially cooked cyprinid fish causes liver (hepatobiliary) disease and cancer (cholangiocarcinoma) due to fishborne zoonotic trematodes (FZT), in particular Clonorchis sinensis and Opisthorchis viverrini. The primary source of transmission is by native fish, but aquaculture fish are also reported to have high infective potential. Here, a cross-sectional survey of FZT in fish farms was conducted in an endemic area in Khon Kaen Province, Thailand. By using conventional and polymerase chain reaction (PCR) methods, we detected O. viverrini and FZT metacercariae (Centrocestus formosanus and Haplorchis taichui) in two popular fish species, Barbonymus gonionotus (silver barb) and Cirrhinus mrigala (mrigal), from aquaculture farms. Both species were infected in five of six farms examined by PCR but not by conventional methods, yet the prevalence of FZT metacercariae in aquaculture fish was high (46.9%). In addition to O. viverrini (17.1%), the native fish Cyclocheilichthys armatus and Hampala dispar had a prevalence of FZT of 81.4%, which included 5.7% for C. formosanus and 17.1% for H. taichui by conventional method. To our knowledge, this is the first discovery of O. viverrini in aquaculture fish in Thailand. More comprehensive studies are required to determine if human-induced disease transmission coupled with natural transmission cycle occurs throughout the aquaculture industry in the region. This has significant impact on food quality and safety, and provides the basis for the development of an effective strategy for the prevention and control of foodborne diseases.

Relationship between snail population density and infection status of snails and fish with zoonotic trematodes in Vietnamese carp nurseries.

BackgroundFish-borne zoonotic trematodes (FZT) are a food safety and health concern in Vietnam. Humans and other final hosts acquire these parasites from eating raw or under-cooked fish with FZT metacercariae. Fish raised in ponds are exposed to cercariae shed by snail hosts that are common in fish farm ponds. Previous risk assessment on FZT transmission in the Red River Delta of Vietnam identified carp nursery ponds as major sites of transmission. In this study, we analyzed the association between snail population density and heterophyid trematode infection in snails with the rate of FZT transmission to juvenile fish raised in carp nurseries.Methodology/Principal FindingsSnail population density and prevalence of trematode (Heterophyidae) infections were determined in 48 carp nurseries producing Rohu juveniles, (Labeo rohita) in the Red River Delta area. Fish samples were examined at 3, 6 and 9 weeks after the juvenile fish were introduced into the ponds. There was a significant positive correlation between prevalence of FZT metacercariae in juvenile fish and density of infected snails. Thus, the odds of infection in juvenile fish were 4.36 and 11.32 times higher for ponds with medium and high density of snails, respectively, compared to ponds where no infected snails were found. Further, the intensity of fish FZT infections increased with the density of infected snails. Interestingly, however, some ponds with no or few infected snails were collected also had high prevalence and intensity of FZT in juvenile fish. This may be due to immigration of cercariae into the pond from external water sources.Conclusions/SignificanceThe total number and density of potential host snails and density of host snails infected with heterophyid trematodes in the aquaculture pond is a useful predictor for infections in juvenile fish, although infection levels in juvenile fish can occur despite low density or absence infected snails. This suggests that intervention programs to control FZT infection of fish should include not only intra-pond snail control, but also include water sources of allochthonous cercariae, i.e. canals supplying water to ponds as well as snail habitats outside the pond such as rice fields and surrounding ponds.

Prevalence and Intensity of Fish-Borne Zoonotic Trematodes in Cultured Freshwater Fish from Rural and Urban Areas of Northern Vietnam

Consumption of raw freshwater fish produced in both rural farm and urban wastewater ponds is a common practice in Vietnam. The present study assessed the risk of fish-borne zoonotic trematode (FZT) infection from fish raised in both these aquaculture systems in northern Vietnam. The diversity, prevalence, and infection intensity of FZT metacercariae in 1,500 freshwater fish collected from 6 sites located in rural and urban areas in northern Vietnam were investigated. The specific diagnosis of species was made by morphologic methods. The overall FZT prevalence in fish from both urban wastewater ponds and rural farm ponds was 11.2%. In wastewater ponds, the overall prevalence was 5.1%, ranging from 2.0% in tilapia to 7.3% in common and grass carp. In fish from farm ponds, the prevalence was 17.3%, and ranged from 6.7% in mud carp to 26.7% in common carp. The mean intensity of FZT infection was also higher in fish from farm ponds than that in fish from wastewater ponds (6.0% and 8.4%, respectively). The FZT species recovered from infected fish included both liver (Clonorchis sinensis) and intestinal flukes (Haplorchis taichui, Haplorchis pumilio, and Centrocestus formosanus). The prevalence of FZT in fish raised in these common farm systems represents a significant public health risk for a population with a strong cultural preference for consuming raw or inadequately prepared fish. These research results should encourage the public health and agriculture sectors to conduct the risk factor research required to develop control programs for FZT.

Raw-Fish-Eating Behavior and Fishborne Zoonotic Trematode Infection in People of Northern Vietnam

Raw fish consumption in restaurants, for example, Sashimi style, is popular worldwide. In Vietnam, raw fish dishes are also traditionally prepared and consumed in private households. However, the habits of eating raw or otherwise inadequately cooked fish can be associated with risks of acquiring fishborne zoonotic trematode (FZT) infection. The present study was done in a fish-farming community in Nam Dinh, Vietnam, to obtain information about habits of eating raw fish dishes and risks for human FZT infection. Discussions were held in different groups divided by gender and age on raw-fish-eating behavior. A total of 180 household members were interviewed and their stool samples analyzed to identify risk factors of FZT infection. There was awareness about the risk of liver fluke infections from eating raw fish. However, many older people accepted these risks and continued eating raw fish, as they know effective drug treatment is available. Raw fish dishes are consumed at social gatherings from shared plates and dipping sauces using the same chop sticks. This is likely to pose risks of crosscontamination with FZT metacercariae to different food items as indicated by the finding that 25.8% of household members that stated not to have eaten raw fish were infected. In total, 32.2% fish farm household members were infected with FZT. The odds of FZT infection was 2.3 times higher (p = 0.013) for those eating raw fish than for those who did not eat raw fish. Among the people eating raw fish, those eating raw fish in restaurants had 3.6 times higher odds of FZT infection (p = 0.009) than people eating raw fish at home. A successful program to control FZT must be based on in-depth knowledge on the social and anthropological determinants of people's raw-fish-eating behavior and hygiene practices as well as production of FZT-free fish for human consumption.

Fish-Borne Zoonotic Trematodes in Cultured and Wild-Caught Freshwater Fish from the Red River Delta, Vietnam

Nam Dinh province in Red River Delta, Vietnam, is an endemic area for the human liver fluke Clonorchis sinensis, but little is known about its occurrence in the fish intermediate host in this region. A cross-sectional study was carried out to identify fish-borne zoonotic trematodes (FZT) species diversity and to estimate the prevalence and infection densities of FZT in three major types of fish: cultured species and self-recruiting species from family ponds and wild-caught species from irrigation canals in Nam Dinh province. A total of 714 wild-caught fish from canals and 829 fish from family ponds were examined for FZT infection. Only a single fish from a pond was infected with Cl. sinensis. The intestinal fluke Haplorchis pumilio was very common and found in more than 50% of fish irrespective of origin. Four other intestinal FZT species were found at low prevalence (<4.0%) except Procerovum varium, which was found in 14.4% of wild-caught fish. There was no significant difference (p > 0.05) in FZT prevalence in cultured fish (64.3%) compared with wild-caught fish (68.9%), nor between cultured fish species (65.1%) and self-recruiting species obtained from ponds (58.1%). The prevalence of FZT in silver carp (Hypophthalmichthys molitrix) (82.7%), grass carp (Ctenopharyngodon idellus) (76.9%), and mrigal (Cirrhinus mrigala) (63.8%) was significantly higher than that in rohu (Labeo rohita) (49.1%) (p < 0.001). The density of FZT metacercariae in fish from canals (0.56 metacercariae/g) was significantly higher (p < 0.001) than that in fish from ponds (0.03 metacercariae/g). The frequent occurrence of intestinal flukes in all fish types indicates that wild-caught and cultured fish are equally at risk of infection with FZT, particularly in species commonly used in Nam Dinh to prepare raw fish dishes. These results underscore the need for greater awareness of the risk from raw fish dishes among public health agencies and consumers.

Fishborne zoonotic trematodes in raw fish dishes served in restaurants in Nam Dinh Province and Hanoi, Vietnam.

Fishborne zoonotic trematode (FZT) parasites are a serious food safety problem in Asian countries because of their tradition of eating raw fish. In northern Vietnam, the prevalence of FZT metacercariae (MC) in wild-caught and cultured fish is quite high. The objective of this study was to assess the risk for acquiring these parasites from raw fish dishes prepared in restaurants. Samples of fish meat (body muscle) and remainder parts (head, gills, fins, skin, and muscle tissue from the tail) were obtained from 39 restaurants located in Nam Dinh province, an area endemic for FZTs, and from 74 restaurants in the capital city of Hanoi. Six species of zoonotic intestinal trematodes were recovered from a total of nine freshwater and brackish water fish species. Overall, the prevalence of MC in meat samples was 6.1% and in remainder samples, 34.9%; freshwater fish were more often infected (39.4%) than brackish water species (16.0%). Dishes from Nam Dinh restaurants were more often infected (11.8%) than those from Hanoi (3.1%). The dominant trematode species, Haplorchis pumilio, exhibited a strong predilection for encystment in remainder fish parts (36.5%) compared with meat (3.9%). The mean density of MC in meat samples overall was low (1.0 to 1.5 MC per 50 g); however, the potential for parasites to accumulate over years of eating raw fish could result in significant clinical disease, as occurs with liver flukes. Also, the high densities of MC in the remainder parts (2.3 to 72.9 MC per 50 g) pose a threat for cross-contamination during dish preparation.

Prevalence of zoonotic trematode parasites in fish fry and juveniles in fish farms of the Mekong Delta, Vietnam

Zoonotic parasites are a significant food safety problem, particularly in Asia. In Vietnam fishborne zoonotic trematodes (FZT) are highly prevalent in fish cultured in grow-out farms. However, FZT infection status of juveniles produced and distributed by hatcheries and nurseries is unknown. Here we report an epidemiological investigation on FZT in fry and juveniles of major cultured freshwater fish species in four provinces of the Mekong Delta of Vietnam. No FZT infections were found in 14 species of fry sampled from hatcheries. In contrast, nursery juveniles of river catfish, hybrid catfish, giant gouramy, climbing perch, common carp, kissing gouramy, silver barb, silver carp, grass carp, Indian carp, pacu, tilapia and snakeskin gouramy were frequently infected with FZT metacercariae (range 1.2–29.7%). Seasonal variation in prevalence was observed: prevalence in river catfish and hybrid catfish were maximal in January, at the end of the flooding season, while the prevalence in juveniles of giant gouramy, climbing perch, common carp, kissing gouramy, silver barb, silver carp grass carp, mrigal and pacu were higher in the wet season, June to November. Overall, FZT prevalence was highest in climbing perch and giant gouramy (29.7% and 27.8%, respectively) and the lowest in river catfish (1.2%). The density of FZT metacercariae in fish varied seasonally only in climbing perch which was maximal in the wet season (P<0.05), compared to the dry season (430 vs 28 metacercariae/100 g of fish). These results demonstrate that acquisition of infected seed stock from nurseries is a serious risk for Vietnamese grow-out fish farms, and stress that interventions to control FZT must focus also on these stages of the cultured fish production cycle.

Prevalence of Zoonotic Trematodes in Fish from a Vietnamese Fish-Farming Community

The prevalence of fish-borne zoonotic trematode (FZT) metacercariae was investigated in fish farmed by rural households in Nghe An Province, located in northern Vietnam. In total, 716 fish, including tilapia (Oreochromis niloticus) and 6 carp species, i.e., grass carp (Ctenopharyngodon idellus), bighead carp (Aristichthys nobilis), mrigal (Cirrhinus mrigala), common carp (Cyprinus carpio), silver carp (Hypophthalmychthys molitrix), and roha (Labeo rohita), collected from 53 fish farms were examined. The overall prevalence of FZT metacercariae was 44.6%, ranging from 12.5% to 61.0% in fish species collected from grow-out ponds, which are the production system for growing fish from fingerling size to market size. The overall prevalence was 43.6% in fingerlings cultured in nurseries, ranging from 7.4% to 62.8% for different fish species. The FZT species recovered were heterophyids and echinostomatids and included Haplorchis pumilio, H. taichui, H. yokogawai, Centrocestus formosanus, Stellantchas musfalcatus, and Echinochasmus japonicus, all of which are intestinal flukes in humans, other mammals, and birds. This is the first report of H. yokogawai and E. japonicus in fish in Vietnam, and the first record for S. falcatus in northern Vietnam. Although a previous cross-sectional survey of the people living in these fish farm households revealed a very low prevalence of FZTs (< 1%), our results demonstrate that intestinal flukes are common in farmed fish in this area, suggesting that reservoir hosts such as dogs, cats, and pigs are more important in sustaining the life cycles of these flukes in fish farms than human hosts. This has implications for the effectiveness of control programs focused mainly on treatment of humans.

Occurrence and species distribution of fishborne zoonotic trematodes in wastewater‐fed aquaculture in northern Vietnam

To assess the risk of fishborne zoonotic trematode (FZT) infection in fish reared in wastewater-fed ponds in peri-urban areas of northern Vietnam. We collected 1200 fish from aquaculture systems receiving wastewater from the cities of Hanoi and Nam Dinh in northern Vietnam and examined them for zoonotic metacercariae. Collection periods were fall (October-November 2005) and spring (April-June 2006). The fish were digested in pepsin and metacecariae, recovered and identified by light microscopy. Data were expressed as both prevalence and intensity of infection. FZT metacercariae were recovered from fish samples at both sites and during both sampling periods. The overall prevalence of FZT was 5% (2.0% in fall and 6.5% in spring) in fish from Hanoi and 4.6% (2.4% in fall and 5.7% in spring) in fish from Nam Dinh. The higher prevalence in the warmer months was likely due to ecological factors such as temperatures and population dynamics of the snail vectors. All metacercariae recovered were zoonotic intestinal trematodes of the family Heterophyidae. The infected fish included tilapia (Oreochromis niloticus) and three species of carps, including silver carp (Hypophthalmichthys molitrix), which is commonly used for the preparation of raw fish dishes. Intensities of infection were relatively low. Fish raised in peri-urban wastewater-fed aquaculture systems are at risk of infection with trematode parasites, which can be transmitted to humans through consumption of raw or improperly prepared fish. However, the prevalence was low as compared to previous findings of FZT in non-wastewater fish elsewhere in Vietnam. Studies are needed to assess the relative importance of different risk factors for transmission and control of FZT in wastewater-fed aquaculture.

Prevalence of fishborne zoonotic parasites in important cultured fish species in the Mekong Delta, Vietnam

A seasonal investigation on the occurrence of fishborne zoonotic trematodes (FZT) in economically important mono-cultured hybrid catfish and giant gouramy was conducted in the Mekong Delta of Vietnam. Fish from carp poly-culture and intensive small-scale integrated vegetable-aquaculture-animal husbandry farming (VAC) systems were also examined. No FZT metacercariae were found in any mono-cultured hybrid catfish. FZT metacercariae were common, however, in fish from the other three systems: All metacercariae belonged to the Heterophyidae family of trematodes, Haplorchis pumilio, H. taichui, Centrocestus formosanus and Stellantchasmus falcatus. The FZT prevalence was 1.7% in mono-cultured giant gouramy, 6.6% in carp from the poly-culture and 3.0% in fish raised in the VAC system. H. pumilio was the most common FZT species constituting more than 58.0% of all metacercariae recovered. The prevalence of infection was significantly higher in the flooding season compared to the non-flooding season for both giant gouramy and fish reared in the VAC system. FZT intensity was greatest in fish from carp poly-culture, particularly in the flooding season. The results indicate that certain fish production systems are at risk for FZT, and control approaches will benefit from understanding these risk factors.