Chuboon, Suksan; Wongsawad, Chalobol; Ruamsuk, Archaree; Nithikathkul, Chusak
Lab-Pla is a famous kind of Thai traditional food in northern and northeasthern Thailand. Consumption of uncooked Lab-Pla can lead to trematode infection because cyprinoid fish serve as the 2nd intermediate hosts of trematodes. The preparation of Lab-Pla can be made in different ways, depending on seasonings used in different places. The effect of the seasonings used in Lab-Pla on the survival of Haplorchis taichui metacercariae were, therefore, determined. The methodology was done by the following: 100 g of chopped Siamese mud carp fish (Henicorhynchus siamensis) were used for each experiment of seasonings used for completely mixed Lab-Pla. Mixed materials were blended and digested by 1% pepsin solution to obtain metacercariae and then checked for the movement under a stereomicroscope. The results showed that metacercariae remained active in 5 experimental groups: fermented fish treated, fish sauce treated, chili powder treated, ground roasted rice treated, and complete mixed treated indicated by average as follows: 27, 38.7, 33.3, 42.7 and 21%, respectively. In conclusion, fermented fish and completely mixed methods have more effectiveness in killing metacercariae in Lab-Pla preparation than other seasonings (p < 0.05).
Wongsawad, Chalobol; Wongsawad, Pheravut
Specific primers for the detection of Opisthorchis viverrini and Haplorchis taichui were investigated by using the HAT-RAPD PCR method. Fourteen arbitrary primers (Operon Technologies) were performed for the generation of polymorphic DNA profiles. The results showed that a 319 bp fragment generated from the OPA-04 primer was expected to be O. viverrini-specific while a 256 bp fragment generated from the OPP-11 primer was considered to be H. taichui-specific. Based on each sequence data, two pairs of specific primers were designed and sequences of each primer were as follows; H. taichui; Hapt_F5'-GGCCAACGCAATCGTCATCC-3'and Hapt_R1 5'-CTCTCGACCTCCTCTAGAAT-3' which yielded a 170 bp PCR product. For O. viverrini, OpV-1F: 5'-AATCGGGCTGCATATTGACCGAT-3' and OpV-1R: 5'-CGGTGTTGCTTATTTTGCAGACAA-3' which generated a 319 bp PCR product. These specific primers were tested for efficacy and specific detection for all parasites DNA samples. The results showed that 170 and 319 bp specific PCR products were generated as equivalent to positive result in H. taichui and O. viverrini, respectively by having no cross-reaction with any parasites tested. PCR conditions are recommended at 68°C annealing temperature and with 0.5 mM magnesium chloride (Mg Cl(2)). Additionally, specific primers developed in this study were effective to determine the presence of both parasites in fish and snail intermediate hosts, which the DNA of O. viverrini was artificially spiked since it is rarely found in northern Thailand. The H. taichui and O. viverrini-specific primers successfully developed in this study can be use for epidemiological monitoring, preventing management and control programs.
Sato, M; Pongvongsa, T; Sanguankiat, S; Yoonuan, T; Kobayashi, J; Boupha, B; Nishimoto, F; Moji, K; Sato, M O; Waikagul, J
The liver fluke, Opisthorchis viverrini, and the minute intestinal fluke, Haplorchis taichui, are prevalent in many Asian countries. This study analysed the patterns of infections of O. viverrini and H. taichui in Lahanam and Thakhamlien villages (Savannakhet Province, Lao PDR), in two cross-sectional investigations. Out of a total of 207 human participants, post-anthelmintic treatment positivity rates for expelled worms were 170 (82.1%) for H. taichui and 65 (31.4%) for O. viverrini. Both these species co-exist in the study villages. When each parasite was analysed separately, H. taichui infections reached a plateau among people aged >20 years. Opisthorchis viverrini infection rates were highest in the age group 21-30 years, with decreasing infection rates after the age of 30. Our findings indicated that fish-borne trematode infections were more prevalent among adults. Fish, common intermediate hosts, were acquired in the study area for analysis. The examination of 35 species of fish as intermediate hosts found O. viverrini metacercariae in only six species, and these were found mostly during the month of November. Many farmers who live on the rice fields obtain their food from their immediate environment, including these intermediate-host fish, potentially putting them at greater risk of O. viverrini infection. By contrast, H. taichui metacercariae were found in three species of fish obtained from the market, meaning that anyone could consume them and become infected. If people who work in rice fields limit the species of fish they consume, or avoid consuming raw fish during the month of November, they may reduce their risk of O. viverrini infection.
Wongsawad, Chalobol; Phalee, Anawat; Noikong, Waraporn; Chuboon, Suksan; Nithikathkul, Choosak
Diseases caused by the liver fluke, Opisthorchis viverrini and the minute intestinal fluke, Haplorchis taichui, are clinically important, especially in the Northeast and North regions of Thailand. It is often difficult to distinguish between these trematode species using morphological methods due to the similarity of their eggs and larval stages both in mixed and co-infections. A sensitive, accurate, and specific detection method of these flukes is required for an effective epidemiological control program. This study aimed to determine the prevalence of O. viverrini and H. taichui infections in human feces by using formalin-ether sedimentation and high annealing temperature random amplified polymorphic DNA (HAT-RAPD) PCR methods. Fecal specimens of people living along the Mae Ping River, Chomtong district were examined seasonally for trematode eggs using a compound microscope. Positive cases were analyzed in HAT-RAPD, DNA profiles were compared with adult stages to determine the actual species infected, and specific DNA markers of each fluke were also screened. Our results showed that out of 316 specimens, 62 were positive for fluke eggs which were pre-identified as O. viverrini and H. taichui. In addition, co-infection among these two fluke species was observed from only two specimens. The prevalence of H. taichui infections peaked in the hot-dry (19.62%), gradually decreased in the rainy (18.18%), and cool-dry seasons (14.54%), respectively. O. viverrini was found only in the hot-dry season (6.54%). For molecular studies, 5 arbitrary primers (Operon Technologies, USA) were individually performed in HAT-RAPD-PCR for the generation of polymorphic DNA profiles. The DNA profiles in all 62 positives cases were the same as those of the adult stage which confirmed our identifications. This study demonstrates the mixed infection of O. viverrini and H. taichui and confirms the extended distribution of O. viverrini in Northern Thailand.
Nissen, Sofie; Nguyen, Lan Anh; Thamsborg, Stig Milan;
Fishborne zoonotic trematodes (FZT) are highly prevalent in Southeast Asia. Recent studies on domestic animal’s role in the transmission of FZT in Northern Vietnam found that the most prevalent FZT was Haplorchis pumilio. The importance of dogs, cats and pigs was assessed, and dogs were found...... to have the highest intensity of infection and contribute the most to the contamination of the environment with FZT eggs in the Nam Dinh province - a highly endemic area for FZTs. Given the free roaming and fish-eating behaviour of many dogs in rural Vietnam controlling the infection in dogs represents......-reared dogs (3-6 months old), were each orally infected with 500 metacercariae (mainly H. pumilio) obtained by artificially digestion of naturally infected fish. Another eight dogs were included as uninfected controls. Faecal examination for eggs was performed twice weekly using a sieving and sedimentation...
Nissen, Sofie; Nguyen, Lan Anh Thi; Dalsgaard, Anders
Fishborne zoonotic trematodes (FZT) are highly prevalent in Southeast Asia. Recent studies on the role of domestic animals in the transmission of FZT in Northern Vietnam found that dogs, mainly infected with Haplorchis pumilio, contributed widely to the transmission of FZT. On this background, we...... conducted an experimental infection with H. pumilio to elucidate population dynamics and host reactions in dogs. Eight household-reared dogs (3-6 months old), were each orally infected with 500 H. pumilio metacercariae obtained by artificial digestion of naturally infected fish. Another eight dogs were...... included as uninfected controls. Faecal examination for eggs was performed twice weekly using a sieving and sedimentation technique. Body temperature and weight of the dogs were measured as was total white blood cells, blood eosinophils and packed cell volume. Subsets of dogs were examined post...
Nissen, Sofie; Nguyen, Lan Anh Thi; Dalsgaard, Anders; Thamsborg, Stig Milan; Johansen, Maria Vang
Fishborne zoonotic trematodes (FZT) are highly prevalent in Southeast Asia. Recent studies on the role of domestic animals in the transmission of FZT in Northern Vietnam found that dogs, mainly infected with Haplorchis pumilio, contributed widely to the transmission of FZT. On this background, we conducted an experimental infection with H. pumilio to elucidate population dynamics and host reactions in dogs. Eight household-reared dogs (3-6 months old), were each orally infected with 500 H. pumilio metacercariae obtained by artificial digestion of naturally infected fish. Another eight dogs were included as uninfected controls. Faecal examination for eggs was performed twice weekly using a sieving and sedimentation technique. Body temperature and weight of the dogs were measured as was total white blood cells, blood eosinophils and packed cell volume. Subsets of dogs were examined post-mortem for presence of adult FZT at three different time points post infection by sectioning of the small intestine and caecum into four parts. Patent infections established in all eight infected dogs. The worm establishment ranged from 15 to 121 flukes (3-24%, mean 12%). Faecal egg excretion was measured in all eight infected dogs but no more than two eggs per g faeces (epg) were found at any time. Infections lasted for at least two months as documented by the presence of adult flukes in all three dogs necropsied on day 58 post infection. The predilection site of the flukes was identified as the lower part of jejunum (93% of total worm burden). The results of the haematological tests did not differ between the infected and uninfected group. Further, no clinical symptoms were observed in the infected group and no macroscopic pathological changes could be assigned to the trematode infections, neither did histopathological examination of the intestine reveal any differences between the infected and the control dogs. This study provides the first basic knowledge on the establishment
Full Text Available nd, and China where endemic animal illness is observed. Infection of humans occurs through the ingestion of ...ong K, Odermatt P, Keiser J, Felger I PCR Diagnosis of Opisthorchis viverrini and Haplorchis taichui Infection
Full Text Available Abstract Background Infections with fish-borne zoonotic trematodes (FZT including the minute intestinal fluke, Haplorchis pumilio, are highly prevalent in Southeast Asia. However, little is known about the infection dynamics and clinical symptoms in the final hosts which include a range of animal species and man. We aimed to generate such information using an experimental model with H. pumilio in foxes. Method Eight commercially bred foxes were each orally infected with 2000 H. pumilio metacercariae. Another three foxes served as uninfected controls. Faecal examination for eggs was performed twice weekly. The body weight was measured, standard haematological and biochemical analysis were performed regularly. All foxes were euthanized at day 56 post infection (p.i.. Adult worms were quantified and location in the small intestine noted. Results Anorexia was observed in all infected foxes starting day 12 p.i. and lasting for approximately a week. A weight loss was noticed in the infected group in weeks 3–6 p.i. Five of eight infected foxes excreted H. pumilio eggs day 9 p.i. onwards, the remaining three started on day 13 p.i. Mean (± SD faecal egg counts showed an initial peak at day 16–20 with a maximum of 1443 ± 1176 eggs per gram of faeces (epg, where after a stable egg output around 4–500 epg was seen. Worm burdens ranged between 116–2070 adult flukes with a mean (± SD worm recovery of 948 ± 666. The majority of worms were found in the lower part of the jejunum. Total white blood cell and lymphocyte counts were significant lower in the infected group from first week p.i. onwards and throughout the study period. A significantly lower level of eosinophils was found in week 2 p.i. and transient anaemia was seen in week 2 and 4 p.i. Conclusion This study showed a short prepatency period, an initial peak in egg excretion, establishment of infection in all animals with predilection site in the lower jejunum and a marked but
Full Text Available Trematode infections are considered to be a non public health problem in Indonesia, with the exception of the two species namely Schistosoma japonicum and Fasciolopsis buski which are still assumed to be the most important trematodes of Indonesia. Other trematode infections reported occasionally were caused by: Echinostoma spp, Haplorchis yokogawai, Haplorchis taichui, Paralecithodendrium molenkampi, Phaneropsolus bonnei and Plachiorchis javensis. Schistosoma japonicum infection has been known to be endemic among the inhabitants of Lake Lindu since 1937. Surveys carried out since that time revealed prevalence rates which varied from 8-55%. Further studies proved that the species at Lindu were similar to the species found in Asia. Resurvey initiated in 1971 gave almost a similar prevalence rate followed by the discovery of the molluscan intermediate host namely Oncomelania hupensis lindoensis. In 1972 a new focus of schistosomiasis was discovered at Napu Valley. Other survey did not reveal any other endemic foci. The result of a biological study carried out since 1976 reconfirmed the diagnosis of the species. Clinical study showed that the significant signs and symptoms of schistosomiasis found among the Lindu inhabitants were : dermatitis, diarrhea, dysentry, abdominal pain, nausea and vomiting, loss of appetite, weakness, distention of the abdomen, melaena. hepatomegaly and splenomegaly. Control of schistosomiasis was initiated in 1981, using praziquantel for mass treatment. Twelve cycles of treatment at Lake Lindu area and 10 cycles at Napu Valley resulted in a reduction of overall prevalence rate from 15,80 % at Lindu area in 1981 to 1,14 % in 1987 whereas in Napu Valley the reduction was from 35,8 % in 1982 to 1,00 % in 1987. Fasciolopsis buski infection was first reported in 1982 from the Regency of Babirik, Hulu Sungai Utara in South Kalimantan Province. The following survey in that area in 1986 revealed a prevalence rate of 27
Li, Kang; Murrell, Kenneth Darwin; Clausen, Jesper Hedegaard
), and Carassius auratus (4).The FZT species recovered were mainly Haplorchis taichui, and H. pumilio along with some unknown species whose identifications are still being determined. Subsequently a cross-sectional survey for the prevalence and diversity of FZT in tilapia culture systems was conducted in Guangdong...... Province during August-October 2011. A total of 389 tilapias sized 2.33-1450.00 g were collected from 30 farms, and examined for FZT metacercariae by the pepsin digestion method. Ten percent of the farms surveyed had fish with FZT infections. The overall FZT prevalence in fish was 1.5% and the infection...
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.
Chuboon, Suksan; Wongsawad, Chalobol
Snail and fish intermediate hosts were collected from rice fields in 3 districts; Mueang, Mae Taeng and Mae Rim of Chiang Mai Province during April-July 2008. For identification of larval trematode infection, standard (cracked for snail and enzymatically digested for fish) and molecular methods were performed. The results showed that three types of cercariae were found, pleurolophocercus, cotylocercous, and echinostome among 4 species of snail with a prevalence of 29, 23 and 3% respectively. Melanoides tuberculata snail was the most susceptible host for cercariae infection. Four species of metacercariae, Haplorchis taichui, Stellantchasmus falcatus, Haplorchoides sp and Centrocestus caninus, were found with a prevalence of 67, 25, 60 and 20%, respectively. The Siamese mud carp (Henicorhynchus siamensis) was the most susceptible fish host for H. taichui, and half- beaked fish (Dermogenys pusillus) for S. falcatus metacercariae infection, whereas Haplorchoides sp and C. caninus were concomitantly found in Puntius brevis. HAT-RAPD profile confirmed that pleurolophocercus cercariae found in Melanoides tuberculata from Mae Taeng District belonged to H. taichui and in Tarebia granifera from Mueang District were S. falcatus.
Pitaksakulrat, Opal; Sithithaworn, Paiboon; Laoprom, Nonglak; Laha, Thewarach; Petney, Trevor N; Andrews, Ross H
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.
El-Azazy, Osama Mohamed ElShfei; Abdou, Nadra-Elwgoud Mohamed Ibrahim; Khalil, Amal Iskander; Al-Batel, Maha Khaled; Majeed, Qais Abdulrazak Habeeb; Henedi, Adawia Abdul-Ruhman; Tahrani, Laila Mohamed Azad
Stray cats are a common feature roaming the streets and alleys of Kuwait; they could be a source of parasites, including trematodes, that affect humans. A survey was conducted to identify feline trematodes and throw the light on their public health significance in Kuwait. Out of 240 stray cats trapped from different localities of Kuwait from June 2011 to May 2012, 59 (24.6%) were found to be infected with 14 species of trematodes. The most common were trematodes of the genus Heterophyes, particularly H. heterophyes and H. dispar that were found in respectively 15.8% and 10.8% of the cats examined. Other trematodes recorded, with lower prevalences, were Heterophyes nocens (2.9%), Haplorchis taichui (3.8%), Stictodora sawakinensis (2.1%), Stellantchasmus falcatus (1.6%), Echinochasmus japonicus (1.6%), and Mesostephanus dottrensi (1.3%). Centrocestus cuspidatus, Galactosomum fregatae, Ascocotyle sp., Mesostephanus appendiculatus, Haplorchis yokogawai, and Pygidiopsis genata showed the lowest prevalence (0.4%) and intensity. The majority of the trematodes are recorded for the first time in Kuwait and even in the Gulf region. The study reveals that stray cats are good indicators of fish-borne trematodes in the environment. As all trematodes recovered are zoonotic, their significance to public health should be considred.
Le, Thanh Hoa; Nguyen, Nga Thi Bich; Truong, Nam Hai; De, Nguyen Van
Mitochondrial DNA sequences offer major advantages over the more usual nuclear targets for loop-mediated isothermal amplification approaches (mito-LAMP) because multiple copies occur in every cell. Four LAMP primers [F3, FIP(F1c+F2), BIP(B1c+B2), and B3] were designed based on the mitochondrial nad1 sequence of Opisthorchis viverrini and used for a highly specific assay (mito-OvLAMP) to distinguish DNA of O. viverrini from that of another opisthorchiid (Clonorchis sinensis) and other trematodes (Haplorchis pumilio, Haplorchis taichui, Fasciola hepatica, and Fasciola gigantica). Conventional PCR was applied using F3/B3 primer pairs to verify the specificity of the primers for O. viverrini DNA templates. All LAMP-positive samples could be detected with the naked eye in sunlight, by gel electrophoresis (stained with ethidium bromide), and by addition of SYBR green I to the product in sunlight or under UV light. Only DNA from O. viverrini yielded amplification products by LAMP (and by PCR verification), and the LAMP limit of detection was as little as 100 fg (10(-4) ng DNA), indicating that this assay is 10 to 100 times more sensitive than PCR. Field testing was done using representative egg and metacercarial samples collected from localities where the fluke is endemic. With the advantages of simplicity, rapidity, sensitivity, and cost effectiveness, mito-OvLAMP is a good tool for molecular detection and epidemiology studies in regions or countries where O. viverrini is endemic, which can lead to more effective control of opisthorchiasis and trematodiasis.
Full Text Available Objective: To analyze the prevalence of trematode infections in the snails intermediate host Tarebia granifera and Melanoides tuberculata collected from Ratchaburi Province by a molecular approach in order to construct a dendrogram to identify species at the cercarial stage. Methods: Specimens were collected from nine districts located in Ratchaburi Province using the stratified sampling method. The samples were examined for cercarial infection using the crushing method. All specimens were amplified by region of internal transcribed spacer 2 (ITS2 based on the PCR technique, and the sequence data were aligned in order to construct the dendrogram by maximum likelihood with 10 000 bootstrap replicates. Results: Five cercarial types were observed comprising parapleurolophocercous cercaria, xiphidiocercaria, megarulous cercaria, furcocercous cercaria and transversotrema cercaria. The overall prevalence of cercarial infection was found to be 7.92% (63 / 795. The parapleurolophocercous cercaria was found in the highest level of prevalence followed by xiphidiocercaria, megarulous cercaria, furcocercous cercaria and transversotrema cercaria, respectively. The parapleurolophocercous cercaria samples were identified and separated into Haplorchis taichui and Haplorchis pumilio, while xiphidiocercaria were identified as Lecithodendriidae and megarulous cercaria were identified as Philophthalmus. The dendrogram separated the cercariae into five groups, which were comprised of parapleurolophocercous cercaria, xiphidiocercaria, megarulous cercaria, furcocercous cercaria and transversotrema cercaria using the sequence of Angiostrongylus cantonensis as an out-group. Conclusions: This is the first study to report on the prevalence of cercarial infection in Ratchaburi Province, Thailand. The high prevalence of cercariae was revealed by the infection in Tarebia granifera and Melanoides tuberculata. Thus, the sequence data of ITS2 can be used to investigate the
Arimatsu, Yuji; Kaewkes, Sasithorn; Laha, Thewarach; Sripa, Banchob
Opisthorchis viverrini and other food-borne trematode infections are major health problems in Thailand, the Lao People's Democratic Republic, Vietnam and Cambodia. Differential diagnosis of O. viverrini based on the microscopic observation of parasite eggs is difficult in areas where Clonorchis sinensis and minute intestinal flukes coexist. Recently, loop-mediated isothermal amplification (LAMP) has been widely used for detection and identification of trematode for its simple method that is useful in low-resource or field settings. We have reported ITS1-LAMP assay to detect O. viverrini infection from human feces. The sensitivity and specificity of the test was 100% and 61.5%. The sensitivity of the test appeared to be higher than microscopic egg examination; however non-specific amplification from other parasites could not be ruled out. We therefore targeted microsatellites of O. viverrini that is a species specific sequence. By using hydroxyl naphthol blue (HNB)-LAMP, O. viverrini microsatellite 6 (OVMS6) could specifically amplify DNA from O. viverrini genome, but not other parasites such as C. sinensis, Opisthorchis felineus, Centrocestus caninus, Haplorchis taichui, Fasciola gigantica and Haplorchoodes sp. The detection limit of the test is 1 ng genomic DNA, which was 1000 times lower than the ITS1-LAMP, but targeting microstellites showed more specific detection of O. viverrini. In addition, the colorimetric LAMP assay was simple and effective; this makes it potentially applicable for point-of-care diagnosis.
Ditrich, O; Giboda, M; Scholz, T; Beer, S A
The egg morphology of the following medically important small flukes from Southeast Asia and Far East were studied: Opisthorchis viverrini, Clonorchis sinensis (Opisthorchiidae), Haplorchis taichui, H. pumilio, H. yokogawai, Stellantchasmus falcatus and Metagonimus sp. (Heterophydiae). This study revealed a great intraspecific variability and interspecific similarity in size and shape of eggs. The eggs shape does not seem to be suitable for species identification. On the other hand, biometrical analysis of egg size enabled us to divide eggs from the species studied into four distinct groups according to the Faust-Meleney index (FMI) characterizing egg size rather than the length and width of eggs. The surface structures of eggs, delineated by using a scanning electron microscope (SEM), appeared to be a suitable morphological feature for distinguishing some groups of small flukes. Eggs from the Haplorchiinae were typified by the characteristic filamentous mesh structure. The problems of identification of eggs in human stool samples and suitability of using morphological criteria such as shape and size of eggs are discussed herein.
Anawat Phalee; Chalobol Wongsawad
Objective:To investigate the infection ofFasciola gigantica(F. gigantica) in domestic cattle from ChiangMai province and molecular confirmation usingITS-2 region.Methods:The liver and gall bladder ofBubalus bubalis(B. bubalis) andBos taurus(B. taurus) from slaughterhouses were examined adult worms and prevalence investigation.The species confirmation with phylogenetic analysis usingITS-2 sequences was performed by maximum likelihood andUPGMA methods. Results:The total prevalences of infection inB. bubalis andBubalus taurus(B. taurus) were 67.27% and52.94% respectively.The respective prevalence in bothB. bubalis andB. taurus were acquired fromDoi-Saket,Muang, andSanpatong districts, with81.25%,62.50% and60.00% for B. bubalis and62.50%,50.00% and47.06% forBos taurus respectively.The species confirmation ofF. gigantica and some related species by basing on maximum likelihood andUPGMA methods used,4 groups of trematodes weregenerated, firstF. gigantica group including specimen ofChiang Mai, second2 samples of F. hepatica, third group of3 rumen flukes;Orthocoelium streptocoelium, F. elongatus andParamphistomum epliclitum and fourth group of3 minute intestinal flukes;Haplorchis taichui, Stellantchasmu falcatus,Haplorchoides sp. and liver fluke;Opisthorchis viverrini respectively.Conclusions:These results can be confirmed theGiant liver fluke which mainly caused fascioliasis inChiangMai was identified asF. gigantica and specimens were the same as those ofF. gigantica recorded in other different countries.Nucleotide sequence of ITS-2 region has been proven as effective diagnostic tool for the identification ofF. gigantica.
Rebecca J Traub
Full Text Available Differentiation of the fish-borne trematodes belonging to the Opisthorchiidae, Heterophyidae and Lecithodendriidae is important from a clinical and epidemiological perspective, yet it is impossible to do using conventional coprological techniques, as the eggs are morphologically similar. Epidemiological investigation therefore currently relies on morphological examination of adult worms following expulsion chemotherapy. A PCR test capable of amplifying a segment of the internal transcribed spacer region of ribosomal DNA for the opisthorchiid and heterophyid flukes eggs taken directly from faeces was developed and evaluated in a rural community in central Thailand. The lowest quantity of DNA that could be amplified from individual adults of Opisthorchis viverrini, Clonorchis sinensis and Haplorchis taichui was estimated at 0.6 pg, 0.8 pg and 3 pg, respectively. The PCR was capable of detecting mixed infection with the aforementioned species of flukes under experimental conditions. A total of 11.6% of individuals in rural communities in Sanamchaikaet district, central Thailand, were positive for 'Opisthorchis-like' eggs in their faeces using conventional parasitological detection techniques. In comparison to microscopy, the PCR yielded a sensitivity and specificity of 71.0% and 76.7%, respectively. Analysis of the microscopy-positive PCR products revealed 64% and 23% of individuals to be infected with O. viverrini and C. sinensis, respectively. The remaining 13% (three individuals were identified as eggs of Didymozoidae, presumably being passed mechanically in the faeces following the ingestion of infected fishes. An immediate finding of this study is the identification and first report of a C. sinensis-endemic community in central Thailand. This extends the known range of this liver fluke in Southeast Asia. The PCR developed herein provides an important tool for the specific identification of liver and intestinal fluke species for future
Li, Kang; Clausen, Jesper Hedegaard; Murrell, Kenneth Darwin
Guangdong province is the most important region for tilapia culture in China. However, it is also an endemic region for fishborne zoonotic trematodes (FZT), which pose a risk to human food safety and health. A study was designed to assess the status of trematode parasite infections in tilapia...... risk of spillover of FZT from area sylvatic reservoir hosts, a survey of local wild-caught fish was conducted. Five species of FZT were discovered from a total of 271 wild-caught fish and a mean infection density of 4.0 metacercariae/100 g; FZT discovered included intestinal flukes (Haplorchis spp...
Mokhtar Ibrahim Khalil
Full Text Available The present study was the first attempt to survey the diversity of fish zoonotic parasites in the southern region of Saudi Arabia, particularly the Najran area, from October 2012 to October 2013. Approximately 163 fish representing seven species (two of freshwater fish and five of marine fish were examined for fish-borne trematode metacercariae using the compression technique, and for zoonotic nematode larvae. Adult flukes were obtained from cats experimentally infected with the metacercariae on day 25 post-infection The prevalence of each parasite species was recorded. The parasites found belonged to two taxa: Digenea (Heterophyes heterophyes and Haplorchis pumilio in muscle tissue; and nematodes (larvae of Capillaria sp. in the digestive tract. The morphological characteristics of the fish-borne trematode metacercariae and their experimentally obtained adults were described. This is the first report of these parasites in fish in Saudi Arabia. Moreover, Myripristis murdjan presented higher prevalence of Capillaria sp. infection (22.7%, while Haplorchis pumilio was the dominant metacercarial species (7.9%. Although the number of documented cases continues to increase, the overall risk of human infection is slight. The increasing exploitation of the marine environment by humans and the tendency to reduce cooking times when preparing seafood products both increase the chances of becoming infected with these parasites. Furthermore, our results indicate that certain fish production systems are at risk of presenting fish zoonotic parasites, and that control approaches will benefit from understanding these risk factors.
Wiriya, Benjamaporn; Clausen, Jesper Hedegaard; Inpankaew, Tawin
was to determine the prevalence of FZT metacercarial infections in Nile tilapia from cage and pond aquaculture systems and in wild-caught fish from Suphan Buri, Nakhon Pathom and Chachoengsao provinces, Thailand. Fish were collected from four cages in Suphan Buri and four ponds in Nakhon Pathom provinces between...... September-October 2011 and April-May 2012 and wild-caught fish were collected in May 2012. All fish were examined for metacercariae by a pepsin digestion and metacercariae identified using morphological and molecular methods. During the first sampling of tilapia the prevalence of metacercariae in cage...... falcatus, Haplorchis pumilio and Procerovum varium. The results revealed a low risk for FZT in Nile tilapia cultured in cage and pond aquaculture systems. However, the high prevalence of FZT in wild-caught fish indicates a high potential for spillover from wild reservoir hosts and underscores the need...
Full Text Available A total of 55 domestic cats (Felis calus f. domestico and one wild (Bengal cat (Prionailurus bengalensis from the Vientiane Province, central Laos, were examined for helminth parasites with emphasis given to potential human parasites. The following species were found (parasites infective to man marked with an asterisk: Opisthorchis viverrini*, Haplorchis pumilio*,H. laichui*,H. yokogawai*, Stellantchasmus falcatus* (Digenea; Spirometra sp.*, Dipylidium caninum*, Taenia taeniaeformis (Cestoda; Capillariidae gen. sp., Toxocara canis*, T. cati*, Ancylostoma ceylanicum*, A. tubaeforme, Gnathostoma spinigerum*, Physaloptera preputials (Nematoda; and Oncicola sp. (Acanthocephala. This study demonstrated that examination of cats may provide useful data on the occurrence of helminths which are potential causative agents of human diseases.
Kanarek, Gerard; Zaleśny, Grzegorz; Sitko, Jiljí; Tkach, Vasyl V
The systematic position and phylogenetic relationships of the family Cortrematidae Yamaguti, 1958 have always been controversial. In the present study, the phylogenetic relationships of this family and its constituent genera and families within the superfamily Microphalloidea were evaluated using previously published and newly obtained sequences of 28S rDNA of Cortrema magnicaudata (Bykhovskaya-Pavlovskaya, 1950) (Cortrematidae), Phaneropsolus praomydis Baer, 1971 and Microtrema barusi Sitko, 2013 (Phaneropsolidae). Results clearly demonstrate that the genus Cortrema Tang, 1951 is closest to Gyrabascus Macy 1935, both genera forming one of the clades within the family Pleurogenidae in the superfamily Microphalloidea and sharing several important morphological features. Thus, the family Cortrematidae should be considered among synonyms of the Pleurogenidae. Based on the analysis of morphology, C. corti Tang, 1951, C. testilobata (Bykhovskaya-Pavlovskaya, 1953) and C. niloticus Ashour, Ahmed et Lewis, 1994 are considered junior synonyms of C. magnicaudata. The phylogenetic position of P. praomydis as a family-level branch not showing close relationships with other families of the Microphalloidea, supports the status of the Phaneropsolidae as an independent family. The genus Parabascus Looss, 1907 previously considered within the Phaneropsolidae clearly belongs to the Pleurogenidae. In addition, the molecular phylogeny has demonstrated that the recently described phaneropsolid Microtrema barusi belongs to the microphallid genus Microphallus Ward, 1901. Therefore, Microtrema Sitko, 2013 is considered a junior synonym of Microphallus. Our analysis has also confirmed the status of Collyriclidae as a family within the Microphalloidea. Not yet sequenced representatives of other families within the Microphalloidea (e.g. Anenterotrematidae, Eumegacetidae, Renschtrematidae, Stomylotrematidae, etc.) need to be included in future molecular phylogenetic studies to better unravel
Thapana Chontananarth; Chalobol Wongsawad
Objective: To investigate the epidemiological situation of cercarial trematodes infection in freshwater snails from different water resources in Chiang Mai province, Thailand. Methods:The snail specimens were collected from 13 districts of Chiang Mai province during April 2008 to February 2012. The prevalence of cercarial infection in snails was investigated using the crushing method. The drawing was done with the help of a camera lucida for the morphological study. Results:A total of 2 479 snail individuals were collected and classified into 7 families, 11 genera, and 14 species, Among them, 8 snails species were found to be infected with an overall prevalence of 17.27% (428/2 479), which infected with nine groups of cercariae;gymnocephalous cercaria, strigea cercaria, megalurous cercaria, monostome cercaria, parapleurolophocercous cercaria (Haplorchis cercaria), pleurolophocercous cercaria, furcocercous cercaria (Transversotrema cercaria), xiphidiocercaria, and virgulate cercaria. The parapleurolophocercous cercaria was found to be the dominant type among the cercarial infection in the snails (64.25%). Conclusions:The various species of snails found in the research location act as the intermediate hosts for the high prevalence of parasitic infection of many species of mammals. This work will provide new information on both the distribution and first intermediate host of trematodes.
Al-Kandari, Wafa Y; Alnaqeeb, Majed A; Isaac, Asha M; Al-Bustan, Suzanne A
Stictodora tridactyla is an intestinal fluke in the family Heterophyidae that parasitizes shorebirds and mammals, including humans. Its metacercarial cyst stage was reported in the Arabian killifish, Aphanius dispar, at Kuwait Bay. In the present study, Cerithidea cingulata was found to serve as the first intermediate host of S. tridactyla. In order to establish the snail-fish link in the life cycle of S. tridactyla, complete sequences of ribosomal DNA internal transcribed spacer region 1 and 2 (rDNA ITS1 and ITS2) and partial sequence of cytochrome oxidase subunit 1 were obtained for metacercarial cysts isolated from the fish A. dispar and rediae isolated from the snail C. cingulata. Sequence alignment demonstrated that these larval stages belong to the same heterophyid species, S. tridactyla. Phylogenetic analysis based on rDNA ITS1, ITS2, and mtCO1 confirmed the position of S. tridactyla within the Heterophyidae and found it to cluster with Haplorchis spp. The present study represents the first molecular study correlating the larval stages of S. tridactyla using rDNA ITS1, ITS2, and mtCO1 and examining the phylogenetic relationships of S. tridactyla with different heterophyid species.
Van Thi Phan
Full Text Available BACKGROUND: Northern Vietnam is an endemic region for fish-borne zoonotic trematodes (FZT, including liver and intestinal flukes. Humans acquire the FZT infection by eating raw or inadequately cooked fish. The production of FZT-free fish in aquaculture is a key component in establishing a sustainable program to prevent and control the FZT transmission to humans. Interventions in aquaculture should be based on knowledge of the main risk factors associated with FZT transmission. METHODOLOGY/PRINCIPAL FINDINGS: A longitudinal study was carried out from June 2006 to May 2007 in Nam Dinh province, Red River Delta to investigate the development and risk factors of FZT infections in freshwater cultured fish. A total of 3820 fish were sampled six times at two-month intervals from 96 fish farms. Logistic analysis with repeated measurements was used to evaluate potential risk factors based on information collected through questionnaire interviews with 61 fish farm owners. The results showed that the FZT infections significantly increased from first sampling in June to July 2006 (65% to sixth sampling in April to May, 2007 (76%. The liver fluke, Clonorchis sinensis and different zoonotic intestinal flukes including Haplochis pumilio, H. taichui, H. yokogawai, Centrocestus formosanus and Procerovum varium were found in sampled fish. Duration of fish cultured (sampling times, mebendazole drug self-medication of household members, presence of snails in the pond, and feeding fish with green vegetation collected outside fish farms all had a significant effect on the development of FZT prevalence in the fish. CONCLUSIONS/SIGNIFICANCE: The FZT prevalence in fish increased by 11 percentage points during a one-year culture period and the risk factors for the development of infection were identified. Results also highlight that the young fish are already highly infected when stocked into the grow-out systems. This knowledge should be incorporated into control
Li, Kang; Clausen, Jesper Hedegaard; Murrell, K Darwin; Liu, Liping; Dalsgaard, Anders
Guangdong province is the most important region for tilapia culture in China. However, it is also an endemic region for fishborne zoonotic trematodes (FZT), which pose a risk to human food safety and health. A study was designed to assess the status of trematode parasite infections in tilapia aquaculture systems as an indicator of potential risks from FZT associated with consumption of tilapia. Tilapia from nursery and grow-out ponds were sampled from monoculture, polyculture and integrated aquaculture systems. The results from 388 tilapia examined revealed a very low prevalence (1.5%) of trematode infections (Heterophyidae and Echinostomatidae). Integrated systems using animal manure and latrine wastes as fertilizer did not show a higher prevalence of FZT. Because it was not clear whether the low risk of infection was attributable to existing effective pond management practices or a low risk of spillover of FZT from area sylvatic reservoir hosts, a survey of local wild-caught fish was conducted. Five species of FZT were discovered from a total of 271 wild-caught fish and a mean infection density of 4.0 metacercariae/100g; FZT discovered included intestinal flukes (Haplorchis spp., Procerovum varium, and Metagonimus spp.) and metacercariae tentatively identified as Clonorchis sinenesis. The common occurrence of FZT in wild-caught fish suggests that the presence of FZT in local wild animal reservoirs is substantial, and that although the current aquaculture management systems for tilapia are generally effective in preventing transmission of these parasites into tilapia production systems, the improvement of pond management practices and biosecurity must be maintained at a high level.
姜进勇; 杜尊伟; 汪丽波; 王学忠; 吴方伟
目的 了解澜沧江下游区域食源性吸虫中间宿主携带囊蚴情况,为云南省食源性吸虫病的防治提供依据.方法 采集的鱼类、溪蟹和青蛙等标本采用人工消化法和水洗沉淀法进行检查. 结果 食源性吸虫中间宿主感染有钩棘单睾吸虫、多刺棘带吸虫、台湾次睾吸虫等囊蚴,集贸市场中南鳢和青蛙的吸虫囊蚴感染率分别为26.56％和3.81％.景洪市勐养镇、勐罕镇、嘎洒镇河流中捕获的南鳢吸虫囊蚴感染率分别为31.60％、77.50％和14.00％. 结论 澜沧江下游某些鱼种、青蛙和溪蟹是食源性吸虫的中间宿主,各级卫生机构应加大食源性吸虫病的健康教育,提高人群防病意识,减少食源性吸虫病.%A survey was conducted to investigate the prevalence of food-borne trematode metacercariae in intermediate hosts in the lower Lancang River basin from Aug. 2009 to July 2010. The survey was done to provide evidence for food-borne trematode prevention and control. Samples of fish, frogs, and crabs were examined by artificial digestion or precipitation and washing. Three species of metacercariae. Haplorchis pumilio, Centrocestus formosanus, and Metorchis orien-talis Tanabc, were mainly detected from the samples. Dwarf snakehead and frogs collected from markets were infected with metacercariae al a respective rate of 26. 56% and 3. 81%. Dwarf snakehead in the town of Mengyang were infected with metacercariae at a rate of 31. 6% , those in the town of Menghan were infected at a rate of 77. 5% , and those in the town of Gasa were infected at a rate of 14. 0%. Results confirmed that some species of freshwater fish and crab play a role as second intermediate hosts of foodborne trematode metacercariae in the lower Lancang River basin. All levels of health facilities should increase health education regarding food-borne trematodes, increase public awareness of disease prevention, and reduce the prevalence of food-borne trematodes.