Morley, Neil J
The presence of trematodes with a free-living metacercarial stage is a common feature of most habitats and includes important species such as Fasciola hepatica, Parorchis acanthus and Zygocotyle lunata. These trematodes encyst on the surface of an animal or plant that can act as a transport host, which form the diet of the target definitive host. Although these species are often considered individually, they display common characteristics in their free-living biology indicating a shared transmission strategy, yet in comparison to species with penetrative cercariae this aspect of their life cycles remains much overlooked. This review integrates the diverse data and presents a novel synthesis of free-living metacercariae using epibiosis as the basis of a new framework to describe the relationship between transport hosts and parasites. All aspects of their biology during the period that they are metabolically independent of a host are considered, from cercarial emergence to metacercarial excystment. Copyright © 2015 Elsevier Ltd. All rights reserved.
Chai, Jong-Yil; Na, Byoung-Kuk; Yong, Tai-Soon; Eom, Keeseon S.; Yoon, Cheong-Ha; Hoang, Eui-Hyug; Jeoung, Hoo-Gn; Socheat, Duong
A survey was performed to investigate the infection status of freshwater fish with zoonotic trematode metacercariae in Phnom Penh and Pursat Province, Cambodia. All collected fish with ice were transferred to our laboratory and examined using the artificial digestion method. In fish from Phnom Penh, 2 kinds of metacercariae (Opisthorchis viverrini and Haplorchis yokogawai) were detected. O. viverrini metacercariae were positive in 37 (50.0%) of 74 fish in 11 species (average no. metacercariae/fish, 18.6). H. yokogawai metacercariae were detected in 23 (57.5%) of 40 fish in 5 species (average no. metacercariae/fish, 21.0). In fish from Pursat Province, 5 kinds of metacercariae (O. viverrini, H. yokogawai, Haplorchis pumilio, Centrocestus formosanus, and Procerovum sp.) were detected; O. viverrini metacercariae (n=3) in 2 fish species (Henicorhynchus lineatus and Puntioplites falcifer), H. yokogawai metacercariae (n=51) in 1 species (P. falcifer), H. pumilio metacercariae (n=476) in 2 species (H. lineatus and Pristolepis fasciata), C. formosanus metacercariae (n=1) in 1 species (H. lineatus), and Procerovum sp. metacercariae (n=63) in 1 species (Anabas testudineus). From the above results, it has been confirmed that various freshwater fish play the role of a second intermediate host for zoonotic trematodes (O. viverrini, H. yokogawai, H. pumilio, C. formosanus, and Procerovum sp.) in Cambodia. PMID:24623879
diplostomatid metacercariae. Parasitology, 89: 209-220. Machado MH, Pavanelli GC and Takemoto. RM 1996 Structure and diversity of endoparasitic infracommunities and the trophic level. Pseudoplatystoma corruscans and. Schizodon borelli. (Osteichthyes) of the High Pavana River. Memorias do Instituto Oswaldo Cruz,.
Rashed, Amr A
Out of 328 specimens of the land snail Monacha obstructa, 105 (32%) were infected with a new metacercaria belonging to the genus Brachylaima Dujardin 1843. The present snail is considered as a new host for this parasite in Egypt. Morphological and morphometric patterns were elucidated. Number of recovered infective metacercariae ranged between 4-35 per kidney of snail host. Metacercariae inhabited only the kidney of infected snails. Total length of mature metacercariae ranged from 1800mu-2050mu (1940mu). The anterior extremity is dome shaped and ceca were highly expanded, undulating, and filled with masses of brown cells and renal tissues. Metacercariae were fully mature, unencyted and highly active. They were either roaming in pathways inside the kidney or firmly anchored to renal tissues via oral sucker. Oral sucker length/total body length is 1:8.5. The histopathological effects involved disintegration of renal lamellae and formation of numerous necrotic masses. Invading metacercariae faced no host resistance and they were surrounded by a parasitovorus vacuole as a mean of immune-evasion. SEM studies revealed that dorsal surface of ventral sucker area is covered with rows of minute spines, Oral suker also is provided with a median tongue-like shaped septum. The rest of integument is aspinose. A critical review for relevant metacercariae of genus Brachylaima is provided.
The identification of Paragonimus spp. lung flukes is based on the morphology of both the metacercaria and the adult. A very small Paragonimus sp. metacercaria was found in a freshwater crab caught in Kanchanaburi Province, West Thailand, an area where metacercariae of Paragonimus heterotremus had not been found. The metacercariae cysts were 180-204 microm in diameter, which was smaller than metacercariae of P. heterotremus. The coefficient of difference in body size between this metacercaria and P. heterotremus was 1.69, which was greater than a subspecific difference. Adults recovered from the lungs of a cat fed with the metacercariae were morphologically similar to, but smaller than, P. heterotremus dissected from the lungs of a feline experimental host. The tegumental spines of the worm in this study were singly spaced in arrangement, which is similar to, but larger than, the spines of P. heterotremus. Therefore, Paragonimus pseudoheterotremus is proposed as a new species.
Kim, Eun-Min; Kim, Jae-Lip; Choi, Sung-Il; Lee, Soon-Hyung
The present study investigated the infection status of Paragonimus westermani metacercariae in freshwater crabs (n = 363) and crayfish (n = 31) from October 2007 to October 2008 using the crush method. All of the freshwater crabs, Eriocheir japonicus, were negative for P. westermani metacercariae while 10 (32.3%) of the 31 examined crayfish were positive. The 10 positive crayfish were caught in Haenam, Jeollanam-do, and there were 8-59 (mean 28.4) metacrcariae per infected crayfish. These results suggest that P. westermani metacerariae are still transmitted by crayfish enzootically in southern Korea, and that freshwater crabs may transmit metacercariae only on rare occasions. PMID:19967096
Vargas-Arzola, Jaime; Segura-Salvador, Aristeo; Reyes-Velasco, Leobardo; Díaz-Chiguer, Dylan L; Márquez-Navarro, Adrián; León-Avila, Gloria; Ibañez-Cervantes, Gabriela; Camacho, Alejandro D; Sánchez-Manzano, Rosa Ma; Nogueda-Torres, Benjamín
Metacercariae of Paragonimus mexicanus were collected in crabs Tehuana guerreroensis (Rathbun, 1933) in the municipality of Putla, Oaxaca, Mexico. Metacercariae were found in 20.8% of the crabs collected, with an average of 1.9 metacercarie per crab. Stained metacercariae showed the specific characteristics of P. mexicanus by morphology and sequencing a fragment of the 28S ribosomal gene obtained by PCR. These findings reveal that T. guerreroensis is an intermediate host for P. mexicanus; this new report is relevant considering the potential risk of transmission in the states of Oaxaca and Guerrero, Mexico. Copyright © 2014 Elsevier B.V. All rights reserved.
Younis, S.A.; Yagi, A.I.; Haroun, E.M.; Gameel, A.A.; Taylor, M.G.
The pathogenesis of unirradiated, 3 krad-irradiated and 20 krad-irradiated metacercarial infections was compared in zebu calves studied over 10 weeks. Calves exposed to 1000 unirradiated metacercariae (mc) became hypoalbuminaemic, and showed elevated serum concentrations of liver enzymes, whereas neither of the other groups was significantly affected. At slaughter, a mean of 332 flukes was recovered from the 0 krad group, while only 23% and 12% of this number were recovered from the 3 krad and 20 krad groups, respectively. All the worms recovered from the 20 krad group were stunted, but a mean of 13% of the flukes recovered from the 3 krad group were large. Liver lesions typical of acute fascioliasis were present in the 0 krad group, but lesions in the other groups, were far less severe. A single vaccination of calves with 10003 krad-irradiated mc induced partial resistance to a challenge with 1000 normal mc eight weeks later, but the reduction in worm recovery was not statistically significant. There was less evidence of protection when two vaccinating doses of 3 krad mc were given within four weeks, with challenge at week 8, and a single vaccination was ineffective against a challenge four weeks later. However, when the irradiation dose was increased to 20 krad, a high level of resistance was induced by a single vaccination, given eight weeks before challenge, and liver pathology was strikingly reduced in the vaccinated animals. (author)
Hughes, D.L.; Doy, T.G. (Agricultural Research Council, Compton (UK). Inst. for Research on Animal Diseases); Hanna, R.E.B. (Queen' s Univ., Belfast, Northern Ireland (UK))
Cattle, sheep and rats were infected orally with ..gamma..-irradiated metacercariae of Fasciola hepatica, or with normal metacercariae. The antibody response was monitored in each host to metacercarial tegument (T0), juvenile tegument (T1), adult tegument (T2) and gut antigens. The response was examined at weekly intervals for cattle and sheep throughout 15 weeks of infection and four weeks after infection in rats, using an indirect fluorescent antibody labelling technique. It was found that the irradiated metacercariae engendered a normal humoral response to T0, T1 and gut antigens in all three hosts although the antibody levels were somewhat reduced due to early death or stunting of the flukes. T0 and T1 appeared to be antigenically similar. Antibodies against T2 appeared late in the animals infected with ..gamma..-irradiated metacercariae and the titres attained were considerably lower than in the controls. The T2 antigen stimulus in the animals given ..gamma..-irradiated metacercariae was probably provided by flukes which 'broke through' the developmental barrier imposed by irradiation and which were found alive at autopsy.
Zhou, Teng-Jian; Chen, Hai-Qiang; Hong, Jia-Lin
To understand the changes of Paragonimus metacercaria infection rates of freshwater crabs in Paragonimus endemic areas and explore the causes in Yongjia County, Zhejiang Province, China. A field investigation was carried out. The freshwater crabs were collected and the metacercaria were separated from the crabs. The infection rates, infectiosities and infection indexes were calculated and the results were vertically compared with the historical findings. The causes of the changes were discussed. Compared with those in 1980, the average infection rate in original endemic areas decreased from 59.71% to 21.50% (P Paragonimus metacercaria infection rate in crabs is lower than before in Yongjia County, but some super high epidemic focus of paragonimiasis still exists. Therefore, we still should strengthen the control measures.
Habe, Shigehisa; Doanh, Pham Ngoc; Yahiro, Shinichiro; Vannavong, Nanthasane; Barennes, Hubert; Odermatt, Peter; Dreyfuss, Gilles; Horii, Yoichiro; Nawa, Yukifumi
Among Paragonimus species, P. paishuihoensis is one of the most mysterious and poorly understood species. Metacercariae are characterized by having a unique dendritically branched excretory bladder. However, the morphology of the adult worm remains unknown. To date, metacercariae of this species have been reported only in China and Thailand. In this study, we first found P. paishuihoensis metacercariae in freshwater crabs, Potamon lipkei, in Hinheub District, Vientiane, Lao PDR, with a prevalence of 77.7% and the average intensity of 10.3 (range 1-28) metacercariae per crab. The molecular data based on ITS2 and CO1 markers indicated that P. paishuihoensis from Laos and Thailand were almost completely identical and were close to members of the Paragonimus bangkokensis/Paragonimus harinasutai complex. Attempts to infect experimental animals (cats, dogs, and rats) with P. paishuihoensis were unsuccessful, suggesting that these animals might be unsuitable definitive hosts for the species. Further studies are necessary to elucidate the taxonomic status and life cycle of P. paishuihoensis.
Uruburu, Mónica; Granada, Mabel; Velásquez, Luz Elena
Paragonimosis or lung fluke disease courses with signs similar to those seen in tuberculosis. The causative agent is a parasite of the genus Paragonimus (Digenea: Troglotrematidae). People become infected by ingesting raw or partially cooked crabs containing metacercariae. The first focus of human paragonimosis in Colombia was recorded in the county of Urrao, where two species of crabs infected with Paragonimus were found. In 2005, crabs with Paragonimus' metacercariae were captured near Medellín, western Colombia. This prompted a search for the parasite in other locations through its presence in the crabs. To establish the distribution of Paragonimus in Antioquia, we evaluated the presence of metacercaria in freshwater brachyuran crabs. From 2005 to 2007, crabs were captured in 13 counties of Antioquia. The crabs were relaxed and dissected to determine presence of trematodes and then to make the taxonomic identifications. From 52 crabs captured in 9 counties, 42 (80.8%) were found with Paragonimus metacercariae. The crabs were identified as Pseudothelphusidae in 2 genera--Hypolobocera and Strengeriana--and were assigned to four species. Three of the species were recorded for the first time as hosts of Paragonimus. A Paragonimus' distribution map was constructed for Antioquia; for the first time urban zones were included. Because of the high rate of infection, the handling and consumption of raw and poorly cooked crabs pose risk factors for human infection. Because crabs are affordable and provide means of easy diagnosis, crabs are targeted as primary agents of and diagnostic tools for paragonmosis.
Hafeez, Md.; Rao, B.V.
Preliminary work has been done to study certain responses induced by irradiated amphistome metacercariae used as a vaccine to immunise lambs, kids and calves. The electrophoretic pattern of the sera collected from lambs and kids vaccinated with gamma irradiated amphistome matacercariae (C.I. XXVI) has been reported in this study. (author). 10 refs., 1 table
Heckmann, R A
Metacercariae of Diplostomum mordax were found in the cranial cavity of Orestias agasii, Orestias olivaceous, Orestias luteus, and Basilichthys bonariensis, fishes from Lake Titicaca, Peru. Metacercariae were not found in Oncorhynchus mykiss introduced into the lake during 1939 and 1940. Compression of neural tissue within and on the surface of the brain was observed in all infected fishes. Metacercariae migrating into the cerebrum and cerebellum of the piscine host caused hemorrhaging, cell necrosis, inflammation, fiber formation, and nerve fiber disruption. The presence of D. mordax in B. bonariensis and the 3 species of Orestias constitute new host records. Infections in the cerebrum and cerebellum add new information on specific parasite location.
Full Text Available Examination of western tubenose goby (Proterorhinus semilunaris from the River Dyje (Czech Republic, Danube basin for metazoan parasites revealed the presence of metacercariae of Cyathocotyle prussica Mühling, 1896 (Digenea: Cyathocotylidae in both muscle tissue and the peritoneal cavity. This is the first time that tubenose gobies have been reported as an intermediate host for C. prussica, and the first time that C. prussica metacercariae have been registered parasitising fish hosts in the Czech Republic. Here, we describe the morphology of metacercariae and five-day preadults, cultivated in vitro, and discuss the importance of recently established, non-native species as suitable hosts for this parasite.
Masala, Simonetta; Piras, Maria Cristina; Sanna, Daria; Chai, Jong-Yil; Jung, Bong-Kwang; Sohn, Woon-Mok; Garippa, Giovanni; Merella, Paolo
This study is a contribution to the molecular taxonomy and epidemiology of heterophyid (Trematoda: Heterophyidae) metacercariae found in the muscle of Mugilidae (Osteichthyes) from Sardinia (western Mediterranean Sea). Sixty specimens of mugilids (13 Chelon labrosus, 18 Liza aurata, 6 Liza ramada, 8 Liza saliens, 15 Mugil cephalus) were examined and 17,899 metacercariae isolated in 95 % of the hosts. Four types of metacercariae were identified: Heterophyes sp. (n = 14,113), Heterophyes sp. -small (1225), Stictodora sp. (1606), and Ascocotyle (Phagicola) sp. (955). The experimental infection of a hamster with Heterophyes sp. metacercariae produced six adults identified as Heterophyes heterophyes and two as Heterophyes cf. nocens. The morphology of Heterophyes sp. -small metacercariae matched with that of Heterophyes dispar. The sequence analysis of the ITS2 and 28S portions of rDNA confirmed the morphological identification of metacercariae, showing four clusters. All adults grouped together with the Heterophyes sp. metacercariae, whereas adults of Heterophyes nocens from Korea clustered separately, showing that this species is distinguished from H. heterophyes, and suggesting caution in the exclusive use of the number of rodlets of the genital sucker to separate the two species. The presence of metacercariae was high in all hosts; the highest prevalence is of Heterophyes sp. (prevalence ≥78 %; mean intensity ≥135 metacercariae/100 g muscle), and the most heavily infected host is M. cephalus (prevalence = 100 %; mean intensity = 841 metacercariae/100 g muscle).
Manuel Tantaleán V.
Full Text Available En los meses de febrero y julio de 1973 se colectaron cangrejos del Valle de Condebamba (Cajamarca correspondientes a Pseudothelphusa chilensis, 69 de ellos (34 machos y 35 hembras fueron examinados, buscando metacercarias de Paragonimus; 22 (64.7% de los machos y 29 (28.85 % de las hembras resultaron infectados con metacercarias, o sea 52 (72.46% de los 69 cangrejos estaban infectados, esta cifra es coincidente con la obtenida por Cuba y cols. (1974.
Acosta, Aline Angelina; Caffara, Monica; Fioravanti, Maria Letizia; Utsunomia, Ricardo; Zago, Aline Cristina; Franceschini, Lidiane; da Silva, Reinaldo José
A complete morphological description, supported by molecular data, of Clinostomum detruncatum metacercariae is reported in this study. The metacercariae were found infecting Synbranchus marmoratus from the Paraná River, municipality of Guaíra, Paraná State, Brazil. The taenioidean uterus is the main morphological characteristic of this species. Both neighbor-joining (NJ) and maximum likelihood (ML) trees showed 2 clear separate clusters grouping together the species from the Palearctic region (Clinostomum complanatum, Clinostomum cutaneum, Clinostomum phalacrocoracis, Clinostomum philippinense) and those from the Nearctic/Neotropical regions (Clinostomum marginatum, Clinostomum tataxumui, C. detruncatum, Clinostomum sp. 7). The latter clade is divided into 2 subclades grouping species from North America and Mexico (C. marginatum and C. tataxumui), and those from Brazil (C. detruncatum and Clinostomum sp. 7).
Caffara, M; Bruni, G; Paoletti, C; Gustinelli, A; Fioravanti, M L
Adults of Clinostomum spp. are digenetic trematodes found in fish-eating birds, reptiles and occasionally mammals, including humans. Freshwater snails serve as first intermediate hosts and many fish species and amphibians as second intermediate hosts. To date, amphibian hosts of Clinostomum metacercariae include members of urodele and anuran families in North America, but no data are available on infections of European amphibians, including newts. In this study, we characterize infections of Clinostomum complanatum metacercariae in four smooth (Lissotriton vulgaris) and 18 Italian crested newts (Triturus carnifex) from an artificial pond located in a protected area in Tuscany, Italy. Parasites were surgically removed from the infected newts and identified both morphologically and using sequences of a mitochondrial gene, cytochrome c oxidase I, and the ribosomal markers, internal transcribed spacers. This is the first record of C. complanatum in European newts and, more generally, in amphibians in Europe.
Yang, Hyun-Jong; Kang, Shin-Yong; Kong, Yoon; Cho, Seung-Yull
The cysteine proteases of Paragonimus westermani metacercariae are involved in metacercarial excystment, host immune modulation, and possibly in tissue penetration. In order to clarify the origin of the enzymes, 28 and 27 kDa cysteine proteases in metacercarial excretory-secretory products were purified through the FPLC system using Mono Q column chromatography. The polyclonal antibodies to the enzymes were produced in BALB/c mice. Immunolocalization studies revealed that both cysteine proteases were distributed at the linings of excretory bladder and excretory concretions of the metacercariae. It was suggested that the excretory epithelium of P. westermani undertake the secretory function of metacercarial cysteine proteases, in addition to its role as a route for eliminating waste products. PMID:12073734
Yang, Hyun-Jong; Chung, Young-Bae; Kang, Shin-Yong; Kong, Yoon; Cho, Seung-Yull
The cysteine proteases of Paragonimus westermani metacercariae are involved in metacercarial excystment, host immune modulation, and possibly in tissue penetration. In order to clarify the origin of the enzymes, 28 and 27 kDa cysteine proteases in metacercarial excretory-secretory products were purified through the FPLC system using Mono Q column chromatography. The polyclonal antibodies to the enzymes were produced in BALB/c mice. Immunolocalization studies revealed that both cysteine protea...
Eosinophils are important effector cells in host defense against parasites. Excretory-secretory product (ESP) produced by helminthic worms plays important roles in the uptake of nutrients, migration in the host tissue, and in immune modulation. However, little is known about the ability of the ESP to directly trigger eosinophil apoptosis. This study investigated whether the ESP of newly excysted metacercariae of Paragonimus westermani could induce apoptosis in human eosinophils. Apoptosis was assayed by staining the cells with FITC-annexin V, and the cells were analyzed by flow cytometry. It was found that the ESP of newly excysted metacercariae of P. westermani induced a direct time- and concentration-dependent increase in the rate of constitutive apoptosis in mature human eosinophils. Eosinophil apoptosis was first apparent 3 hr after treatment with the ESP and continued to increase after 6 hr of incubation with respect to the cells cultured in the absence of the ESP. While only 2.8% of the eosinophils incubated in the medium for 3 hr were apoptotic, 7.6%, 10.9% and 22.6% of the eosinophils treated with 10, 30 and 100 µg/ml ESP were apoptotic, respectively. This result suggests that the ESP of newly excysted metacercariae of P. westermani directly induce eosinophil apoptosis, which may be important for the survival of the parasites and the reduction of eosinophilic inflammation in vivo. PMID:10743354
Fernádez, María Virginia; Hamann, Monika Inés; Kehr, Arturo Ignacio
In Argentina, no ecological studies have been reported on the infection parameters of Kalipharynx sp. metacercariae in planorbid snails. To this end, the aims of this study were: (i) to provide information on the population biology of Kalipharynx sp. metacercariae in the planorbid snails Biomphalaria tenagophila and B. occidentalis through the study of prevalence and intensity of larval infection during a seasonal cycle; (ii) to evaluate the effects of host shell size on prevalence and infection intensity, (iii) to evaluate the effect of infection intensity on cyst size. Samples were taken between June 2010 and April 2011 (encompassing all seasons) from a subtropical permanent pond in Corrientes City, Corrientes, Argentina. A total of 362 metacercariae (n=262 and n=100; from B. tenagophila and B. occidentalis respectively) were collected from 616 snails (n=466 and n=150 from B. tenagophila and B. occidentalis respectively). The metacercarial cysts were found in the digestive gland, mantle cavity, intestine and ovotestis. B. tenagophila showed a range of infection from 1 to 60 cysts per snail (mean = 4.5 +/- SD=9.9), and cyst diameter ranging between 255 and 705 microm (466.4 +/- 119); while, B. occidentalis showed a range of infection from 1 to 23 (5.5 +/- 5.6), and cyst diameter ranging between 310 to 900 microm (554.5 +/- 150). Results obtained indicated that, although absent in autumn, metacercariae of Kalipharynx sp. were present most of the year in both species of Biomphalaria, showing high values in both warm-season. Furthermore, both the infection intensity and host shell size varied significantly between seasons, although no seasonal variation was observed with respect to metacercarial cyst size, suggesting the possibility of more than one peak of cercariae emergence during the year. The prevalence of infection was significantly and positively correlated with snail size in both host species (p 0.05). The results of this study show a significant influence of
Metacercariae of Renifer heterocoelium (Trematoda: Reniferidae in tadpoles of Rhinella schneideri (Anura: Bufonidae in Brazil Metacercarias de Renifer heterocoelium (Trematoda: Reniferidae en renacuajos de Rhinella schneideri (Anura: Bufonidae en Brasil
Hudson Alves Pinto
Full Text Available Renifer heterocoelium is a parasite of the oral cavity of Neotropical snakes for which larval stages and the life cycle are still unknown. During studies of parasites of Rhinella schneideri, tadpoles collected in a lake from the municipality of Santa Luzia, Minas Gerais, Brazil, yielded specimens with metacercariae adhered to the external intestinal wall. After morphological characterization, the metacercariae were identified as R. heterocoelium. This is the first record and morphological characterization of metacercariae of R. heterocoelium for Brazil.Renifer heterocoelium es un parásito de la cavidad bucal de las serpientes neotropicales cuyas formas larvales y ciclo de vida son todavía desconocidos. En un estudio de los parásitos de renacuajos de Rhinella schneideri recogidos en un estanque ubicado en el municipio de Santa Luzia, Minas Gerais, Brasil, se encontraron ejemplares con metacercarias en la pared intestinal. Después de caracterizarlos morfológicamente, los quistes fueron identificados como R. heterocoelium. Este es el primer registro y caracterización morfológica de las metacercarias de esta especie en Brasil.
Hughes, D.L.; Doy, T.G.
Cattle, sheep and rats were infected orally with γ-irradiated metacercariae of Fasciola hepatica, or with normal metacercariae. The antibody response was monitored in each host to metacercarial tegument (T0), juvenile tegument (T1), adult tegument (T2) and gut antigens. The response was examined at weekly intervals for cattle and sheep throughout 15 weeks of infection and four weeks after infection in rats, using an indirect fluorescent antibody labelling technique. It was found that the irradiated metacercariae engendered a normal humoral response to T0, T1 and gut antigens in all three hosts although the antibody levels were somewhat reduced due to early death or stunting of the flukes. T0 and T1 appeared to be antigenically similar. Antibodies against T2 appeared late in the animals infected with γ-irradiated metacercariae and the titres attained were considerably lower than in the controls. The T2 antigen stimulus in the animals given γ-irradiated metacercariae was probably provided by flukes which 'broke through' the developmental barrier imposed by irradiation and which were found alive at autopsy. (author)
Full Text Available This studyw as carried out to establish whether cattle can develop resistance to re-infectionby Calicophoron microbothrium by assessing the response of intestinal mucosal globule leukocytese, osinophils, mast cells and basophils, and the establishment of the parasite in the host. A total of 241-year old Tuli steers were randomly divided into four groups of six animals each and infected with C. microbothriumm etacercariae. On the first day of the study, animals in Groups I and II were immunized with 5000 metacercariae and then challenged with 15000 metacercariae on Day 150 post immunization. Animals in Group III were immunized with 15000 metacercariae at the same time that Groups I and II animals were challenged to act as a positive control group Animals in Group IV were left uninfected and acted as a negative control group. Three animals from each group were slaughtered on Day 28 post-challenge and the remainder were slaughtered on Day 42 post-challenge. The established amphistomes were recovered and histopathological and cytological examinations were done on the jejunum, duodenuma, bomasum and the rumen. The establishment rates of the challenge infection in the immunized and challenged groups were lower and ranged from 0 to 0.2% as compared to 6% from naive animals infected as positive controls. Animals immunized and then challenged with C. microbothrium had significantly higher eosinophil, mast cell and globule leukocytes counts in the intestinal mucosa (P < 0.05 as compared to those of the control group. The study indicates that cattle can develop resistance to C. microbothrium re-infection and that eosinophils and mast cells may be important cells in the rejection of the parasite.
Kim, Min-Ki; Pyo, Kyoung-Ho; Hwang, Young-Sang; Chun, Hyang Sook; Park, Ki Hwan; Ko, Seong-Hee; Chai, Jong-Yil; Shin, Eun-Hee
Artificial digestive solution based on pepsin is essential for collecting metacercariae from fish. To promote the enzymatic reactivity of pepsin, the pH of the solution has to be adjusted to pH 1.0-2.0. Hydrochloride (HCl) is usually used for this purpose, but the use of HCl raises safety concerns. The aim of this work was to address the usefulness of citric acid as an alternative for HCl for the acidification of pepsin solution, and to examine its potential to damage metacercariae during in vitro digestion as compared with HCl. Changes in pH after adding 1-9% of citric acid (m/v) to pepsin solution were compared to a 1% HCl (v/v) addition. Digestion of fish muscle was evaluated by measuring released protein concentrations by spectrophotometry. In addition, survival rates of metacercariae in pepsin solution were determined at different citric acid concentrations and were compared that of with 1% HCl. The present study shows that addition of citric acid reduced the pH of pepsin solutions to the required level. Addition of more than 5% of citric acid resulted in the effective digestion of fish muscle over 3h in vitro, and 5% citric acid was less lethal to metacercariae than 1% HCl in pepsin solution. Pepsin solution containing 5% citric acid had digestive capacity superior to pepsin solution containing 1% HCl after 3h incubation with released protein concentrations of 12.0 ng/ml for 5% citric acid and 9.6 ng/ml for 1% HCl. Accordingly, the present study suggests that the addition of 5% citric acid to pepsin solution is a good alternative to 1% HCl in infection studies because citric acid is a stable at room temperature and has a good safety profile. In addition, we suggest that the use of citric acid enables the preparation of commercial digestive solutions for the detection of microorganisms in fish and other vertebrate muscle tissue. Copyright © 2013 The Authors. Published by Elsevier B.V. All rights reserved.
Cuba, César; Ibáñez H., Nicanor; Fernández, Eduardo; Miranda, Hernán
Se ha estudiado el índice de infección natural y la intensidad del parasitismo en cangrejos recolectados en tres provincias del Departamento de Cajamarca, Perú, habiéndose encontrado metacercarias de Paragonimus Braun, 1899 en el 43.97% de 539 especímenes de Pseudothelphusa chilensis Milne Edwards, 1843, único cangrejo hallado en la zona. El órgano parasitado fue casi exclusivamente el hepatopancreas, siendo 10.33 el número promedio de metacercarias por cangrejo. Las formas adultos logradas m...
Singh, T Shantikumar; Sugiyama, Hiromu; Devi, K Ranjana; Singh, L Deben; Binchai, Sutheewan; Rangsiruji, Achariya
This study was aimed to find out the host-parasite relationship between Paragonimus heterotremus isolated as metacercariae from mountain crabs, Indochinamon manipurensis, in Manipur, India and laboratory animals such as puppies, albino rats, Swiss mice, guinea pigs, and rabbits, as experimental animals. The animals were fed with the metacercariae. Infected animals were sacrificed 35 to 430 days after feeding to recover worms, which were used to determine the developmental stages. Adult worms (n = 14) were recovered from 3 puppies > or = 70 days after feeding and immature worms (n = 25) were recovered from 2 other puppies 35 or 43 days after infection. The infection rate in puppies was 100%. Juvenile worms were recovered from 3 of 13 rats: 1 of 11 rats whose viscera and cavities were examined and both of two rats whose muscles were examined. Rats were not a suitable animal model for pulmonary infection with P. heterotremus. Mice, guinea pigs, and rabbits were also found to be insusceptible to pulmonary infection with P. heterotremus.
Esmey B.E. Moema
Full Text Available The metacercarial (larval stages of diplostomid digeneans are known to inhabit freshwater fish, causing tissue damage in the process. Due to their widespread diversity, little is known about their life cycle. The classification of these parasitic stages to the species level using only the morphology is very challenging due to the lack of genitalia; they are regarded to be the most important structures in the identification of these organisms. In this study, additional morphological information through light and scanning electron microscopy is given for two different diplostomids found in the cranial cavity of Clarias gariepinus and the vitreous chambers of Tilapia sparrmanii and Pseudocrenilabrus philander. The diplostomid metacercaria inhabiting the cranial cavity of Clarias gariepinus was morphologically identified as Diplostomulum (Tylodelphys mashonenseand an unknown metacercaria of the genus Diplostomumwas found in the vitreous chambers of Pseudocrenilabrus philander and Tilapia sparrmanii. Both parasitic species’ 28S recombinant deoxyribonucleic acid genomic regions were successfully amplified using Dig 125/1500R primer pairs. The assay yielded a product of approximately 1300 base pairs as seen on the gel images. There were 14 nucleotide differences over the entire analysed sequences resulting in a 1.1% (14/1273 nucleotide difference. In line with the morphological characteristics of these parasites, there seemed to be a slight difference in their genetic makeup. The application of molecular techniques on digenetic trematodes seems very promising and may yield great potential in future descriptions of morphologically similar parasitic species.
During the period from October 1996 to November 1998, the infection status of Paragonimus westermani metacercariae in freshwater crayfish (Cambaroides similis) collected from Bogildo (islet), Wando-gun, Chollanam-do, which is known for an endemic area of P. westermani in Korea, were examined. The average infection rate of Paragonimus metacercariae in crayfish was 88.6%, and mean number of metacercariae per infected crayfish was 30.2. This metacercarial density was the highest in the group of weight in 7.1-9.0 g. These results suggest that the natural life cycle of P. westermani is still well-preserved in Bogildo. PMID:10188385
Arifin, M.; Boky, J.T.; Tarmizi
An experiment was carried out to study the effect of irradiation against to the infectivity of F.gigantica by using observation the exchange of goat bodies condition. Four groups of experiment animals were inoculated by irradiated meta cercaria of F. gigantica at a doses of 0, 45, 55 and 65Gy, and the other one is the negative control. Each experiment animal received 350 lives meta cercaria. The infectivity of irradiated meta cercaria in goat has been followed in the development of body weight, blood value described as the number of red blood cells (RBC), level of haemoglobin (Hb), percentages of packed cell volume (PCV), eosinofil and the last is pathology anatomic inspection. The results obtained showed that 45 Gy is the optimal dose of irradiation for decreasing infectivity of metacercariae of F.gigantica which have the ability to stimulate the good immune response in the goat
Full Text Available Clinostomidae are digeneans characterized by a complex taxonomic history, continuously under revision based on both morphological and molecular analysis. Among the 14 species considered valid so far Clinostomum phalacrocoracis has been well described only at the adult stage, whereas the morphology of the metacercarial stage has been reported only once. During a parasitological survey carried out on 262 wild cichlids sampled from Lake Kinneret (Israel metacercariae referable to C. phalacrocoracis were found in 18 fingerlings. In this study, we report this clinostomid species for the first time in wild fish from Israel describing the metacercarial stage of Clinostomum phalacrocoracis, coupling its morphological description with molecular analysis carried out on ITS rDNA and COI mtDNA sequences.
Clausen, Jesper Hedegaard; Madsen, Henry; Murrell, Kenneth Darwin
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......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...... 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...
Sohn, Woon-Mok; Yong, Tai-Soon; Eom, Keeseon S; Sinuon, Muth; Jeoung, Hoo-Gn; Chai, Jong-Yil
The metacercariae of Artyfechinostomum malayanum (Leiper, 1911) Mendheim, 1943 were discovered in Pila sp. snails purchased from a market in Phnom Penh, Cambodia. They were isolated from the snails using the artificial digestion technique and were orally fed to 2 hamsters, 1 rat, and 2 mice to obtain the adult flukes. The metacercariae were round, 145-165 μm in diameter, having a cyst wall of 6-10 μm in thickness, a head collar and collar spines, and characteristic features of excretory granules. Adult flukes were recovered in the small intestines of the animals at days 14 and 32 post infection and were morphologically observed using a light microscope and a scanning electron microscope. They were plump or elongated, ventrally curved, 6.0-8.1 × 1.6-2.0 mm in size, and characterized by the head collar bearing 43 collar spines, including 5 end group ones on each side, a long cirrus sac extending beyond the posterior margin of the ventral sucker, a submedian ovary, and 2 deeply lobed testes. Eggs in uteri were operculate, ovoid to ellipsoid, and 120-135 × 68-75 μm in size. In scanning electron microscopy, the head collar was prominent with collar spines looking like horns. Scale-like tegumental spines were densely distributed on the ventral surface between the head collar and ventral sucker. Sensory papillae were distributed mainly on the tegument around suckers. By this study, it has been first confirmed that the life cycle of A. malayanum exists in Cambodia.
Doanh, Pham Ngoc; Shinohara, Akio; Horii, Yoichiro; Habe, Shigehisa; Nawa, Yukifumi; The, Dang Tat; Le, Nguyen Thi
Paragonimosis is an important food-borne zoonosis especially in Asian countries. Among Paragonimus species, Paragonimus westermani followed by P. skrjabini complex are the major pathogens for human paragonimosis in Asia. In addition, P. heterotremus is an important pathogen in southern China and the Indochina Peninsula and is the only proven species to cause human paragonimosis in Vietnam. During a recent survey in Yenbai Province in northern Vietnam, we found small and large types of Paragonimus metacercariae often concurrently in mountainous crabs, Potamiscus tannanti. Adult worms from those small and large metacercariae were obtained separately by experimental infection in dogs and cats. Morphological and molecular phylogenetic study based on sequences of ITS2 and a part of CO1 genes were performed for the identification of small and large metacercariae and their adults. The results showed that small metacercariae and their adults are completely identical with P. heterotremus in morphology and molecular genetic profiles. In contrast, large metacercariae and their adults have some morphological similarities with P. skrjabini and P. harinasutai, but are unidentifiable from each other by morphology alone. Molecular phylogenetic tree analyses on ITS2 and CO1 genes revealed that large metacercariae and their adults were grouped in the same clade and different from any known Paragonimus species. Although they share the same ancestor with P. skrjabini complex, their genetic distance was considerably different from two other known subspecies, P. skrjabini skrjabini and P. skrjabini miyazakii. Our results provide a new insight on the phylogeny of the genus Paragonimus.
Alina Huiza F
Full Text Available Un hámster hembra adulta infectada oralmente con 11 metacercarias de Paragonimus mexicanus (peruvianus murió a los 10 días de ser infectado. A la autopsia del animal, se observaron y recuperaron 5 ejemplares juveniles de Paragonimus mexicanus (peruvianus aún móviles: 2 en tejido pulmonar, 2 en cavidad toráxica y 1 en tejido graso del peritoneo abdominal.
Metacercarias del tipo Paramonilicaecum (Digenea: Didymozoidae, parásitos accidentales de elasmobranquios (Elasmobranchii del golfo de México e identificación de metacercarias (Didymozoidae de la Colección Nacional de Helmintos Metacercariae of the type Paramonilicaecum (Digenea: Didymozoidae, accidental parasites in elasmobranchs (Elasmobranchii from the Mexican Gulf of Mexico and identification of metacercarias (Didymozoidae of the Colección Nacional de Helmintos
Full Text Available Como parte de un estudio sobre los helmintos parásitos de rayas (Elasmobranchii, se colectaron 12 ejemplares en 3 localidades del estado de Veracruz, México: Laguna de Tamiahua, Laguna de Tampamachoco y un arrecife de la costa del golfo de México. Se encontraron metacercarias identificadas como parásitos accidentales del tipo Paramonilicaecum (Digenea: Didymozoidae en la válvula espiral de Dasyatis say (raya látigo chata, Gymnura micrura (raya cola de rata y Rhinobatus lentiginosus (guitarra diablito. De manera complementaria, se actualizó la identificación de material perteneciente a Didymozoidae depositado en la Colección Nacional de Helmintos en el Instituto de Biología, Universidad Nacional Autónoma de México. Éste es el primer registro para México de metacercarias del tipo Paramonilicaecum en rayas.During an ongoing study of the helminths of stingrays (Elasmobranchii, 12 stingrays were collected from 3 localities in the state of Veracruz, Mexico: Laguna de Tamiahua, Laguna de Tampamachoco, and a costal reef in the Gulf of Mexico. Metacercarias were collected from the spiral valves of Dasyatis say (bluntnose stingray, Gymnura micrura (lesser butterfly ray, and Rhinobatus lentiginosus (Atlantic guitarfish. The metacercarias, accidental parasites, were assigned to type Paramonilicaecum (Digenea: Didymozoidae. Material (Didymozoidae previously deposited in the Colección Nacional de Helmintos, Instituto de Biología, Universidad Nacional Autónoma de México, was re-identified. This is the first report of a species in larval phase of the "Type" Paramonilicaecum from the Gulf of Mexico, Veracruz, and the first report from a Mexican stingray.
Hernández-Chea, Roderico; Jiménez-Rocha, Ana Eugenia; Castro, Ruth; Blair, David; Dolz, Gaby
The trematode Paragonimus mexicanus is the etiological agent of paragonimiasis, a food-borne zoonotic disease in Latin America. This species, as well as Paragonimus caliensis, have been reported from Costa Rica, but it is not known if the two are synonymous. Two types of Paragonimus metacercariae from freshwater pseudothelphusid crabs from several localities in Costa Rica were recognized by light microscopy. Morphologically, these corresponded to descriptions of P. mexicanus and P. caliensis. Metacercariae of the former species lacked a membrane or cyst and their bodies were yellow in color. Those of P. caliensis were contained in a transparent thin cyst and were pink in color. Morphotypes of metacercariae were determined using scanning electron microscopy (SEM). Based on the number and distribution of papillae in the ventral sucker, three morphotypes were found for P. mexicanus and two for P. caliensis. Analysis of DNA sequences (nuclear ribosomal 28S and ITS2 genes, and partial mitochondrial cox1 gene) confirmed the presence of P. mexicanus and provided the first molecular data for P. caliensis. The two species are phylogenetically distinct from each other and distant from the Asian species. The confirmation of P. caliensis as a separate species from P. mexicanus raises several questions about the ecology, biological diversity, and epidemiology of the genus Paragonimus in Costa Rica. Copyright © 2016 Elsevier Ireland Ltd. All rights reserved.
Song, Jin-Ho; Dai, Fuhong; Bai, Xuelian; Kim, Tae-Im; Yang, Hyun-Jong; Kim, Tong-Soo; Cho, Shin-Hyung; Hong, Sung-Jong
An epidemiological study was performed to know the recent infection status of Paragonimus westermani metacercariae (PwMc) in freshwater crayfish, Cambaroides similis, from 2 streams in Jeollanam-do, Republic of Korea. Crayfish were collected from creeks in Bogil-do (Island), Wando-gun, and in a creek near Daeheung Temple in Haenam-gun. The infection rate of crayfish with PwMc in Bogil-do was 89.8%, and the metacercarial burden was 37 PwMc per the infected crayfish. Crayfish in a creek near Daeheung Temple were larger and twice heavier than those in Bogil-do. Of them, 96.5% were infected with PwMc. An average of 140 metacercariae was found in the infected crayfish, almost quadruple to those of Bogil-do. There was a strong correlation between the number of PwMc and body weight of the crayfish. These results suggest that P. westermani metacercariae are still prevalent in crayfish of the 2 regions in Jeollanam-do, Korea. PMID:28719962
de Montaudouin, Xavier; Blanchet, Hugues; Desclaux-Marchand, Céline; Lavesque, Nicolas; Bachelet, Guy
The European cockle (Cerastoderma edule) is an exploited bivalve along the North-Eastern Atlantic semi-sheltered ecosystems. Its population dynamics are driven by numerous environmental factors, including parasitism, particularly by trematodes. The complex life cycle of trematodes includes a free living stage (cercaria) between the first (a mollusk) and the second (an invertebrate or a vertebrate) intermediate hosts. Taking the example of Himasthla quissetensis, a dominant trematode in Arcachon Bay (France) utilizing Nassarius reticulatus as first intermediate host and the cockle as second intermediate host, we investigated the correlation between cockle infection and cercariae emergence by N. reticulatus, including the role of water temperature and light. Experimental and field data provided evidence that cercariae emergence from N. reticulatus took place during the night and for water temperature comprised between 15 °C and 22 °C (optimum at 20 °C). The lifespan of cercariae did not exceed 48 h and 50% of them were no more infectious after 7 h. A model was designed (emergence = f(water temperature)) in order to calculate an average monthly cercarial emergence rate according to water temperature that was daily recorded for 5 years (1998-2002). Concomitantly, non-infected cockles were transplanted each of these years in order to assess the infection rate by H. quissetensis. Infection occurred strictly for the same water temperatures (and dates) as those expected for cercariae emergence. Within each year, there was often (but not always) a good correlation between the predicted number of emerging cercariae and the observed number of cercariae transformed into metacercariae in cockles. However, the among-years variability of cockle infection (× 4.4 range) was not explained by the predicted annual number of emerging cercariae. This study highlights the importance of water temperature and light in stimulating cercariae emergence which is closely related to the
Kim, Tae Im; Oh, Se-Ra; Dai, Fuhong; Yang, Hyun-Jong; Ha, Sang-Do; Hong, Sung-Jong
Soy sauce-marinated freshwater crabs (Eriocheir japonicus) are a source of human paragonimiasis. The viability of Paragonimus westermani metacercariae (PwMc) in marinated crabs was investigated in an experimental setting. The PwMc collected from freshwater crayfish were inoculated into freshwater crabs, which were then frozen or marinated in soy sauce. All PwMc in the freshwater crabs were inactivated after freezing for 48 h at -20 °C and after freezing for 12 h at -40 °C. After marinating for 32 days, the survival rate of PwMc in 5% NaCl soy sauce was 50%, in 7.5% NaCl soy sauce it was 33.3%, and in 10.0% NaCl soy sauce it was 31.3%. When marinated for 64 days, all PwMc were inactivated in all experimental groups. These results revealed that freezing and soy sauce marination were detrimental to the survival of PwMc in freshwater crabs. Specifically, freezing crabs for more than 48 h or soaking them in soy sauce containing at least 5.0% NaCl for 64 days can inactivate PwMc. These results can inform the production of the traditional Korean soy sauce-marinated freshwater crabs known as gejang.
Sohn, Woon-Mok; Kim, Deok-Gyu; Jung, Bong-Kwang; Cho, Jaeeun; Chai, Jong-Yil
Pygidiopsis cambodiensis n. sp. is described based on adult flukes recovered from Syrian golden hamsters experimentally infected with metacercariae from mullets (Liza macrolepis) purchased at a local fish market in Phnom Penh, Cambodia. The specimens were examined by light and scanning electron microscopy. Among the 13 species so far assigned to Pygidiopsis, the new species belongs to the summa-type (including Pygidiopsis pelecani, Pygidiopsis phalacrocoracis, Pygidiopsis piclaumoreli, Pygidiopsis plana, and Pygidiopsis summa) which lack circumoral spines and have vitelline follicles extending posteriorly from the level of the ovary some distance into the post-testicular space and the uterus not exceeding the acetabulum anteriorly. The new species differs from the other five species of the summa-type particularly in the morphology of the ventrogenital complex, including the genital sac, gonotyl, and gonotyl spines (= rodlets). The genital sac is well developed, sucker-like, slightly larger than the ventral sucker, muscular, and equipped with two gonotyls on the ventral side of the sac. Gonotyls are protruding pad-like, and the number of rodlets on the left gonotyl is four to five and that on the right gonotyl is 10-11 in two rows. This is the fifth Pygidiopsis species reported in Asia, following P. summa (Japan, Korea, and Vietnam), P. phalacrocorasis (Japan), P. pelecani (China), and Pygidiopsis marivillai (Philippines).
André Carrara Morandini
Full Text Available Metacercaria specimens of the genus Opechona (Trematoda: Digenea: Lepocreadiidae are described parasitizing "coelenterates" (scyphomedusae and ctenophores from Southeastern Brazil (São Paulo state. The worms are compared to other Opechona species occurring on the Brazilian coast, but no association has been made because only adult forms of these species have been described. Suppositions as to the possible transference of the parasites are made.Exemplares de metacercárias do gênero Opechona (Trematoda: Digenea: Lepocreadiidae são descritos parasitando "celenterados" (cifomedusas e ctenóforos no sudeste do Brasil (estado de São Paulo. Os vermes foram comparados a outras espécies de Opechona ocorrentes no litoral brasileiro, porém nenhuma associação foi realizada devido às demais espécies terem sido descritas a partir de exemplares adultos. São apresentadas suposições sobre as possíveis formas de transferência dos parasitas.
Bonett, Ronald M; Steffen, Michael A; Trujano-Alvarez, Ana L; Martin, Samuel D; Bursey, Charles R; McAllister, Chris T
Land-use alterations can have profound influences on faunal distributions, including host-parasite relationships. Yellow grub trematodes ( Clinostomum spp.) have complex life cycles involving 3 hosts: a snail, a fish or amphibian, and a bird. Here, we analyze the distribution, prevalence, intensity, abundance, and genetic diversity of encysting metacercariae of Clinostomum spp. in salamanders and fishes throughout an aquatic system that includes a natural Ozark stream and man-made ponds. We found Clinostomum sp. infecting permanently aquatic Oklahoma salamanders ( Eurycea tynerensis ; 56% prevalence) and larval grotto salamanders ( Eurycea spelaea ) immediately downstream from a man-made pond. However, Clinostomum sp. did not infect any salamanders in the spring that supplies this pond, or in sections farther downstream (~0.5 and 2 km). Metacercariae of Clinostomum sp. were present in ~90% of introduced largemouth bass ( Micropterus salmoides ) in the man-made pond adjunct to the stream. Morphological examination and phylogenetic analyses based on the mitochondrial gene cytochrome oxidase 1 ( Co1 ) and the nuclear ribosomal gene 18S show that fishes and salamanders at this site are primarily infected with Clinostomum marginatum . There is a relatively high degree of mitochondrial haplotype diversity in C. marginatum at this site but no consistent genetic difference between parasites in largemouth bass from the man-made pond and those in salamanders from the stream. Based on the microgeographic distribution and relationships of metacercariae of C. marginatum at this site, we hypothesize that the adjunct man-made pond has created an ecological situation that brings the cercariae of this parasite into contact with novel stream salamander hosts.
Full Text Available Se ha estudiado el índice de infección natural y la intensidad del parasitismo en cangrejos recolectados en tres provincias del Departamento de Cajamarca, Perú, habiéndose encontrado metacercarias de Paragonimus Braun, 1899 en el 43.97% de 539 especímenes de Pseudothelphusa chilensis Milne Edwards, 1843, único cangrejo hallado en la zona. El órgano parasitado fue casi exclusivamente el hepatopancreas, siendo 10.33 el número promedio de metacercarias por cangrejo. Las formas adultos logradas mediante inoculación de las metacercarias, fueron identificadas como Paragonimus peruvianus Miyazaki, Ibáñez y Miranda, 1969, con excepción de tres ejemplares que correspondieron a lo especie Paragonimus caliensis Little, 1968.
Rizvi, A; Zaidi, Z A; Alam, M M; Zafar, A; Shareef, P A A; Saifullah, M K; Saleemuddin, M; Abidi, S M A
Clinostomum complanatum is a digenetic trematode that causes yellow grub disease in some fish species and also shows zoonotic potential by sporadically infecting humans. In this study, progenetic metacercariae of C. complanatum were obtained from the fish Trichogaster fasciatus, and were aseptically placed in conjunctival incisions made in the superior and inferior fornices of the eye of rabbits, which served as the experimental hosts. Worms were harvested without necropsy of the host on days 4 and 8 post infection, to observe in vivo transformation of the progenetic metacercariae into ovigerous adult worms. The worms appeared to cause minimal damage to the host although they were tenaciously attached. In vivo maturation was evident by the development of the vitellaria, enlargement of gonads, the presence of a large number of shelled eggs in a distended uterus and ramifications of the intestinal caeca. Obtaining mature ovigerous worms without sacrificing the host clearly gives the rabbit eye model an advantage over those described previously. Due to the relative advantage of the short time required for maturation and the prolific egg production by C. complanatum, it is suggested that this host-parasite system could be used as an excellent model for classroom teaching of trematode biology and to investigate the cues involved in in vivo transformation and host-parasite interactions.
Petrov, A; Podvyaznaya, I
Recent confocal microscopy studies have greatly expanded our knowledge of muscle systems in cercariae and adult digeneans, but the gross anatomy and development of metacercarial musculature remain relatively little known. To further our understanding of metacercarial development, this study used phalloidin staining and confocal microscopy to examine changes in muscle architecture over the course of development from cercariae to infective metacercariae in Diplostomum pseudospathaceum Niewiadomska, 1984. The paper describes muscle development in the body wall, anterior organ (oral sucker), acetabulum, pharynx and midgut and in the musculo-glandular organs that first appear in metacercariae (lappets and holdfast). The muscle architecture of the cercarial tail is also described. The results of the study support previously reported observations that diplostomid musculature undergoes substantial transformation during metacercarial development. The most profound changes, involving extensive remodelling and replacement of cercarial muscles, were seen in the body-wall musculature and in the anterior organ as it developed into the oral sucker. Muscle systems of other cercarial organs showed more gradual changes. The adaptive importance of developmental changes in musculature is discussed.
Tandon, V; Prasad, P K; Chatterjee, A; Bhutia, P T
In several mountainous regions of Northeastern India, foci of Paragonimus infection reportedly involving species that are known to prevail in China have been identified. The present study was undertaken to demonstrate the surface fine topography and sequence analysis of the ribosomal deoxyribonucleic acid (rDNA; second internal transcribed spacer, ITS2) of the metacercarial stages of the lung fluke collected from a mountain stream of the area (Miao, Changlang District in Arunachal Pradesh). The encysted metacercariae were oval in shape and had a smooth surface. The newly excysted metacercaria had a ventral sucker larger than the oral; the body surface was covered with numerous single-pointed and thorn-like tegumentary spines, of which those on the anterior part of the body were bigger in size and showed a gradual reduction in length and number towards the posterior end; dome-shaped papillae in variable numbers were seen around the rim of the oral sucker and were sparsely distributed all over the body surface. The polymerase chain reaction-amplified rDNA ITS2 sequences of the metacercariae were aligned with known sequences for the various species of Paragonimus, and the expectation value was found to be most significant with P. westermani, revealing an absolute match. The surface topography including the number and distribution of papillae and spination patterns and the ITS2 sequences of the metacercariae strongly suggest that the Paragonimus species, prevalent in the region of India, is in fact P. westermani.
The inhibitory effects of L-type Ca2+ channel antagonists on Na cholate-induced in vitro excystment (CIIE) of Paragonimus ohirai metacercariae were studied. At concentrations of 10 microM, nicardipine and nimodipine inhibited CIIE completely and by approximately 92%, respectively. Nitrendipine and (+/-)-verapamil inhibited CIIE by about one half and one third, respectively. Nifedipine and diltiazem did not inhibit CIIE significantly. At higher concentrations, nitrendipine at 20 microM completely inhibited CIIE, and (+/-)-verapamil at 40 microM inhibited CIIE by 93%. Nifedipine and diltiazem inhibited CIIE only slightly and little, respectively, even at 40 microM. Complete inhibition by nicardipine at 10 microM required preincubation of metacercariae with the antagonist for 15 min. The inhibitory effects of nicardipine and nimodipine were reversible, and most of the nimodipine-treated metacercariae could excyst within 1 h after being washed, but the nicardipine-treated ones started to excyst 1 h after washing. Nicardipine suppressed the active movement of encysted juveniles evoked by Na cholate, whereas nimodipine did not suppress this significantly. These results suggested that L-type Ca2+ channels appeared to be involved in CIIE of P. ohirai metacercariae and that the inhibitory effect of the channels was due primarily to factors other than the inhibition of muscular activity, probably involving the secretion and release of enzymes lytic against the metacercarial cyst wall.
Doanh, Pham Ngoc; Hien, Hoang Van; Nonaka, Nariaki; Horii, Yoichiro; Nawa, Yukifumi
During our epidemiological surveys for Paragonimus species in central Viet Nam, we found four morphologically different Paragonimus metacercariae in mountainous crabs. They were identified as metacercariae of Paragonimus westermani, P. bangkokensis, P. proliferus, and P. harinasutai in the order of their prevalence in crab hosts. This is the first discovery of P. harinasutai in Viet Nam, co-inhabiting with P. bangkokensis and other species. Metacercariae of P. harinasutai were given orally to a cat to obtain adult worms. Then, ITS2 and CO1 sequences of metacercariae and adults of P. harinasutai, and metacercariae of P. bangkokensis collected from the same place were determined for analyses of phylogenetic relationships to other P. harinasutai and P. bangkokensis populations as well as related species. The results of molecular analyses showed that P. harinasutai from Quang Binh province of central Viet Nam was almost completely identical with those from Vientiane, Lao PDR; P. bangkokensis from Quang Binh, Viet Nam was also almost completely identical with those from Lao PDR and from Quang Ninh province, Viet Nam. Except for one P. harinasutai isolate from China, all populations of P. harinasutai and P. bangkokensis from Thailand, Lao and Viet Nam make a single clade in both ITS2 and CO1 trees. In ITS2 sequences, AT deletion and ATC insertion were observed in some isolates of both species, indicating recent gene flow between P. harinasutai and P. bangkokensis. Moreover, because of their extremely high genetic similarities and their co-inhabitation in the same crab hosts found in Thailand, Lao PDR and Viet Nam, they should be considered as the sister species at the early stage of divergence. In addition, P. microrchis previously described from Yunnan, China should be placed as the synonym of P. harinasutai, because of their morphological and molecular similarities. Copyright © 2012 Elsevier Ireland Ltd. All rights reserved.
Alina Huiza F.
Full Text Available Cangrejos de rio Hypolobocera chilensis eigenmanni fueron colectados de acequias durante el año 1997 en estación seca (mayo a diciembre en Chaquicocha, Área que pertenece al distrito de Condebamba (departamento de Cajamarca en la parte norte del Perú. Ciento treinta y un cangrejos colectados fueron transportados al Laboratorio de Parasitología y examinados por disección, 27 de 131 (20,6% estaban infectados por metacercarias de Paragonimus mexicanus(=peruvianus. La intensidad de la infección fue de 1 a 5 en la mayoría de los; casos (81,5% con un promedio de 4,85 por cangrejo. Estos datos son diferentes; a los de estudios anteriores; en la misma Área donde fueron más; altos, lo que indica una tendencia al decrecimiento del número de cangrejos infectados.
Bakhmet, Igor; Nikolaev, Kirill; Levakin, Ivan
Trematode parasites can affect their molluscan hosts, which serve as the first intermediate hosts in their life cycles, in manifold ways, but little is known about trematode-induced effects on their second intermediate hosts. Experimental infection of blue mussels Mytilus edulis serving as second intermediate hosts for larval stages (metacercariae) of the trematodes Himasthla elongata was studied in field experiments during one year. The heart rates and growth rates of noninfected mussels were significantly higher than those of infected mussels. During the summer, the heart rates of noninfected mussels showed rhythmic oscillations, whereas the parasitized animals displayed no any rhythmicity. There was a significant difference between the infected and uninfected mussels in relation to heart rates and temperature. The results indicate that mussels infected with H. elongata metacercariae may be at an energetic disadvantage relative to noninfected mussels. Furthermore, trematode infection may disrupt neuronal control of cardiac function.
Song, Jin-Ho; Dai, Fuhong; Bai, Xuelian; Kim, Tae-Im; Yang, Hyun-Jong; Kim, Tong-Soo; Cho, Shin-Hyung; Hong, Sung-Jong
An epidemiological study was performed to know the recent infection status of Paragonimus westermani metacercariae (PwMc) in freshwater crayfish, Cambaroides similis , from 2 streams in Jeollanam-do, Republic of Korea. Crayfish were collected from creeks in Bogil-do (Island), Wando-gun, and in a creek near Daeheung Temple in Haenam-gun. The infection rate of crayfish with PwMc in Bogil-do was 89.8%, and the metacercarial burden was 37 PwMc per the infected crayfish. Crayfish in a creek near Daeheung Temple were larger and twice heavier than those in Bogil-do. Of them, 96.5% were infected with PwMc. An average of 140 metacercariae was found in the infected crayfish, almost quadruple to those of Bogil-do. There was a strong correlation between the number of PwMc and body weight of the crayfish. These results suggest that P. westermani metacercariae are still prevalent in crayfish of the 2 regions in Jeollanam-do, Korea.
It is well known that the cysteine proteases in excretory-secretory product (ESP) of Paragonimus westermani newly excysted metacercariae (PwNEM) are capable of degrading IgG in vitro. Recent evidence suggests that the IgG-coated surface, such as found on parasites, is one of the most effective physiologic stimuli for granulocyte activation. Therefore, this study was designed to investigate the effect of excretory-secretory product (ESP) of PwNEM on superoxide production of granulocytes stimulated with IgG. The 96-well plates were coated with human IgG (0, 10, 30, 100 µg/ml) in the absence or presence of ESP. When granulocytes were incubated in the wells coated with human IgG in the presence of ESP, the level of superoxide production of granulocytes was reduced to about 90% when compared to the cells incubated in the wells coated with IgG alone. This inhibitory effect of the ESP on IgG-induced superoxide production of granulocytes was concentration-dependent. These results suggest that ESP secreted by PwNEM may be important in the control of effector functions of granulocytes stimulated with IgG in human paragonimiasis. PMID:10905073
Full Text Available Se determinó el grado de infección de los crustáceos Ptychophallus tristani presentes en una área endémica de Paragonimus mexicanus, encontrándose que de un total de 182 cangrejos, 161 resultaron positivos (88,5% por metacercarias. Al comparar la infección entre machos y hembras no se encontró diferencias estadísticamente significativas y en relación a su tamaño se observó una relación directa entre tamaño y porcentaje y grado de infección. Se estableció que no existen diferencias significativas en relación a la época del año en que los cangrejos fueron capturados lo que epidemiológicamente es importante pues indica que las fuentes de infección permanecen potencialmente activas durante todo el año.
Luz Elena Velásquez
Full Text Available Introducción. La paragonimosis, o distomatosis pulmonar, es una enfermedad con sintomatología similar a la observada en la tuberculosis. Es causada por parásitos del género Paragonimus (Digenea: Troglotrematidae. Las personas se infectan al consumir cangrejos crudos o mal cocidos, con metacercarias del parásito. El primer foco de paragonimosis humana en Colombia se registró durante 1995 en Urrao, Antioquia, donde se hallaron dos especies de cangrejos que hospedaban el parásito. En el 2005 se capturaron cangrejos con metacercarias de Paragonimus en Medellín, lo que motivó la búsqueda del parásito en otras localidades, mediante su presencia en estos crustáceos. Objetivo. Establecer la distribución de Paragonimus en Antioquia, evaluando la presencia de metacercarias en macrocrustáceos braquiuros, dulciacuícolas. Materiales y métodos. Desde 2005 hasta 2007 se capturaron cangrejos en 13 municipios antioqueños. Se relajaron y sacrificaron para la búsqueda del digeneo y la identificación taxonómica. Resultados. En nueve municipios se capturaron 52 cangrejos, 42 (80,76% con metacercarias de Paragonimus. Todos los crustáceos se determinaron como Pseudothelphusidae, de los géneros Hypolobocera y Strengeriana, y se asignaron a cuatro especies. Tres se registran por primera vez como huéspedes del parásito. Conclusión. Se inicia la construcción de un mapa con la distribución de Paragonimus en Antioquia que incluye por primera vez zonas urbanizadas. Se ratifican el consumo y la manipulación de los cangrejos crudos y mal cocidos como factores de riesgo para la infección humana. Se propone a los cangrejos como agentes focalizadores de paragonimosis por ser asequibles y de fácil diagnóstico.
Gállego, Laia; Gracenea, Mercedes
Cornu aspersum is a terrestrial edible snail, often parasitized by Brachylaima (Trematoda) metacercariae. Ingestion of undercooked snails by humans allows metacercariae to develop to adult in the intestine causing brachylaimiasis (expected mortality rate 5-10%). The cosmopolitan character of the trematode and of its vector, enhanced in a future climate change scenario, and the absence of adequate sanitary controls of the snails in marketplaces clearly increase the risk of human brachylaimiasis. The treatment of farmed snails with praziquatel (PZQ) would be a tool to control this food-borne disease. The objectives of this study were: to report the prevalence of Brachylaima metacercariae parasitizing C. aspersum in marketplaces, to assess the efficacy and tolerance of PZQ in C. aspersum and to develop an HPLC-MS/MS analytical method to quantify PZQ residue in the edible snail (the acceptable daily intake of PZQ is 0.17 mg/kg bw in humans). Prevalence of parasitization of C. aspersum by Brachylaima metacercariae in public marketplaces reached the 80%. PZQ was orally administered to snails, mixed with the usual snail food. In dose determination assays three doses were individually tested (10 days): 1.2 mg PZQ/snail, 1.8 mg PZQ/snail (efficacy 97.3%, p < 0.001) and 2.4 mg PZQ/snail (efficacy 98.7%, p < 0.001). In dose confirmation tests (n = 200) the 2.4 mg PZQ/snail dose was group tested (10 snails/box, 7 days): 2 g of PZQ supplemented snail food were disposed daily in the treatment group boxes and 2 g of snail food (placebo) in the control boxes (efficacy 94.6%, p < 0.001; prevalence dropped from 68.7% in control group to 10.1% in treatment group, p < 0.001). Received dose was 220.2 mg PZQ/kg snail with shell. In the analytical method, linearity, lower limit of quantification (0.05 μg/ml), selectivity, carry over, accuracy, precision, dilution integrity, matrix effect and stability were tested. Sixty snails were treated (11 mg PZQ/g snail food
Full Text Available Echinostome metacercariae are the infective stage for humans and animals. The identification of echinostomes has been based until recently on morphology but molecular techniques using sequences of ribosomal RNA and mitochondrial DNA have indicated major clades within the group. In this study we have used the ITS2 region of ribosomal RNA and the ND1 region of mitochondrial DNA to identify metacercariae from snails collected from eight well-separated sites from an area of 4000 km2 in Lamphun Province, Thailand. The derived sequences have been compared to those collected from elsewhere and have been deposited in the nucleotide databases. There were two aims of this study; firstly, to determine the species of echinostome present in an endemic area, and secondly, to assess the intra-specific genetic diversity, as this may be informative with regard to the potential for the development of anthelmintic resistance and with regard to the spread of infection by the definitive hosts. Our results indicate that the most prevalent species are most closely related to E. revolutum, E. trivolvis, E. robustum, E. malayanum and Euparyphium albuferensis. Some sites harbour several species and within a site there could be considerable intra-species genetic diversity. There is no significant geographical structuring within this area. Although the molecular techniques used in this study allowed the assignment of the samples to clades within defined species, however, within these groupings there were significant differences indicating that cryptic speciation may have occurred. The degree of genetic diversity present would suggest the use of targeted regimes designed to minimise the selection of anthelmintic resistance. The apparent lack of geographic structuring is consistent with the transmission of the parasites by the avian hosts.
Comparative study of the sensibility of Metacercaria cysts of Phagicola Faust, 1920 (Trematoda: Heterophyidae) to ionizing radiation and to freezing in raw mullets Mugil Linnaeus, 1758 (Pisces: Mugilidae)
Moraes, Ivany Rodrigues de
Foodborne illnesses have been a growing concern to the governments, international organizations and consumers, mainly regarding the damages they cause to human health. This issue is the result of the substitution of imported food habits, which enable the consumption of raw fish or wrongly prepared fish. Among the emergent parasitic zoonosis, the Fagicolose stands out. It is transmitted by Phagicola longa, a trematoda of great importance in public health and responsible for high percentages of Mugilidae infections, that is able to produce humans' parasitism. In this experiment, the studies were focused to the radio sensibility of metacercaria of Phagicola longa exposed to a treatment with ionizing radiation (gamma rays and electron beams) and the sensibility to freezing, in artificially infected fish. Some data were acquired regarding the occurrence of Phagicola longa in mullets bought in the local market, the freezer temperature of 10 domestic refrigerators, and the origin and preservation of the mullets in some Japanese-type restaurants in Sao Paulo. The results indicated more effectiveness in the treatment with the electron accelerator, as well as with the freezing at the temperature of-16 deg C. (author)
Onsurathum, Sudarat; Pinlaor, Porntip; Haonon, Ornuma; Chaidee, Apisit; Charoensuk, Lakhanawan; Intuyod, Kitti; Boonmars, Thidarut; Laummaunwai, Porntip; Pinlaor, Somchai
Contamination of a popular fermented fish dish, pla-som, by Opisthorchis viverrini metacercariae (OVMC) is a possible cause of carcinogenic liver fluke infection in Thailand. Affected individuals are at risk of bile duct cancer, which is a major health problem for people in the Greater Mekong Subregion. In order to investigate concerns about food safety, we studied the effects of fermentation time and low temperature on the viability and infectivity of OVMC during the pla-som production process. Pla-som was prepared at room temperature for up to 1 week in duplicate experiments using cyprinid freshwater fish obtained from an O. viverrini-endemic area. OVMC were then isolated and identified under a stereomicroscope. Complete and viable OVMC were found on days 1-4 of fermentation, while their morphology was degenerated thereafter. After OVMC were fed to hamsters, the percentage of the worm recovery after 1 to 2 months of infection was 52%, 44.7%, 11.3% and 1% for days 1, 2, 3 and 4, respectively. In order to measure the effect of low temperature on OVMC, fish were kept in a refrigerator (4 °C) for up to five days and then subsequently fermented for three days. In fish stored in a refrigerator for 1 and 2 days, viable OVMC were clearly observed and were able to infect hamsters, a worm-recovery percentage of 3.3% and 12.7%, respectively. By contrast, in pla-som prepared from fish stored for 3 to 5 days, OVMC were degenerated and could not infect the host. In conclusion, pla-som fermentation for more than four days and refrigerating fish for three days before pla-som processing can prevent O. viverrini infection. This study may increase awareness of fermented-fish dish preparation to prevent liver fluke infection. Copyright © 2015 Elsevier B.V. All rights reserved.
Kesik-Brodacka, Malgorzata; Lipiec, Agnieszka; Kozak Ljunggren, Monika; Jedlina, Luiza; Miedzinska, Katarzyna; Mikolajczak, Magdalena; Plucienniczak, Andrzej; Legocki, Andrzej B; Wedrychowicz, Halina
Cysteine proteinases of Fasciola hepatica are important candidates for vaccine antigens because of their role in fluke biology and host-parasite relationships. In our previous experiments, we found that a recombinant cysteine proteinase cloned from adult F. hepatica (CPFhW) can protect rats against liver fluke infections when it is administered intramuscularly or intranasally in the form of cDNA. We also observed considerable protection upon challenge following mucosal vaccination with inclusion bodies containing recombinant CPFhW produced in Escherichia coli. In this study, we explore oral vaccination, which may be the desired method of delivery and is potentially capable of preventing infections at the site of helminth entry. To provide antigen encapsulation and to protect the vaccine antigen from degradation in the intestinal tract, transgenic plant-based systems are used. In the present study, we aimed to evaluate the protective ability of mucosal vaccinations of 12-week-old rats with CPFhW produced in a transgenic-plant-based system. To avoid inducing tolerance and to maximise the immune response induced by oral immunisation, we used the hepatitis B virus (HBV) core protein (HBcAg) as a carrier. Animals were immunised with two doses of the antigen and challenged with 25 or 30 metacercariae of F. hepatica. We obtained substantial protection after oral administration of the plant-produced hybrids of CPFhW and HBcAg. The highest level of protection (65.4%) was observed in animals immunised with transgenic plants expressing the mature CPFhW enzyme flanked by Gly-rich linkers and inserted into c/e1 epitope of truncated HBcAg. The immunised rats showed clear IgG1 and IgM responses to CPFhW for 4 consecutive weeks after the challenge.
Comparative study of the sensibility of Metacercaria cysts of Phagicola Faust, 1920 (Trematoda: Heterophyidae) to ionizing radiation and to freezing in raw mullets Mugil Linnaeus, 1758 (Pisces: Mugilidae); Estudo comparativo da sensibilidade de cistos de Metacercarias de Phagicola Faust, 1920 (Trematoda: Heterophyidae) a radiacao ionizante e ao congelamento em peixes crus preparados a partir de tainha Mugil Linnaeus, 1758 (Pisces: Mugilidae)
Moraes, Ivany Rodrigues de
Foodborne illnesses have been a growing concern to the governments, international organizations and consumers, mainly regarding the damages they cause to human health. This issue is the result of the substitution of imported food habits, which enable the consumption of raw fish or wrongly prepared fish. Among the emergent parasitic zoonosis, the Fagicolose stands out. It is transmitted by Phagicola longa, a trematoda of great importance in public health and responsible for high percentages of Mugilidae infections, that is able to produce humans' parasitism. In this experiment, the studies were focused to the radio sensibility of metacercaria of Phagicola longa exposed to a treatment with ionizing radiation (gamma rays and electron beams) and the sensibility to freezing, in artificially infected fish. Some data were acquired regarding the occurrence of Phagicola longa in mullets bought in the local market, the freezer temperature of 10 domestic refrigerators, and the origin and preservation of the mullets in some Japanese-type restaurants in Sao Paulo. The results indicated more effectiveness in the treatment with the electron accelerator, as well as with the freezing at the temperature of-16 deg C. (author)
Durieux, Eric D. H.; Galois, Robert; Bégout, Marie-Laure; Sasal, Pierre; Lagardère, Françoise
Little attention has been paid to flatfish parasitism as a potential factor of variation in year-class strength. The aim of this study was to evaluate the relationship between parasitic infection and nutritional condition of young-of-year (YOY) common sole. Sole were collected monthly from May (early settlement) to November 2004 in a mussel pole culture area, using a push-net or a trawl, and dissected for parasite identification and counts. Total lipids were extracted from the whole body and lipid classes quantified. Triacylglycerols to sterols (free form) ratio (TAG/ST) was used as an index of nutritional status. Digenean metacercariae infection, characterized by two dominant genera, displayed clear seasonal dynamics and the highest records yet reported for sole. Prevalence (P%) and mean abundance (Ab ± SE) increased for Timoniella spp. (Acanthostomidae) from May (P% = 22%; Ab = 1.9 ± 1.1) to August (P% = 100%; Ab = 51.2 ± 10.4), and for Prosorhynchus spp. (Bucephalidae) from June (P% = 8%; Ab = 0.1 ± 0.0) to August (P% = 100%; Ab = 19.5 ± 1.7). Epidemiological values reached a plateau after August. In this shellfish-dominated nursery area, YOY sole accumulated metacercariae in the presence of the first intermediate hosts which live in the same habitat. Total lipids decreased significantly from May to June and then remained constant until autumn, indicating a change in the proximate composition of YOY during this high growth period. Mean (± SD) TAG/ST ratio increased from very low values in May-July (0.10 on average) to a peak in September (0.57 ± 0.31) and then fell back to values as low as before. YOY sole only stored energy as TAG prior to autumn. No relationship was found between YOY parasitic infection levels and lipid condition indices. However, the poor nutritional status and heavy infection level observed in November could dramatically lower over-winter survival of the YOY remaining in this shallow part of the nursery.
Metacercárias de Neascus sp. em Geophagus brasiliensis (Perciformes: Cichlidae do rio do Peixe, Juiz de Fora, Brasil - DOI: 10.4025/actascibiolsci.v30i3.493 Neascus-type metacercariae in Geophagus brasiliensis (Perciformes: Cichlidae of Peixe river, Juiz de Fora, Brazil - DOI: 10.4025/actascibiolsci.v30i3.493
José Luis Luque
Full Text Available A metacercária de Neascus sp., causadora da doença dos pontos pretos, é descrita em espécimes de Geophagus brasiliensis coletados no Brasil. Entre março e maio de 2006, foram capturados 33 espécimes de G. brasiliensis, coletados com o auxílio de vara e anzol no Rio do Peixe, Monte Verde, município de Juiz de Fora, Estado de Minas Gerias. Dos 33 espécimes examinados, 30 (90,9% apresentaram-se parasitados por metacercárias causadoras dos pontos pretos, com intensidade média de infecção de 15,6 ± 16,1 (1 – 75. Foi observada correlação significativa entre o comprimento total de G. brasiliensis e a abundância parasitária (r = 0,459; P Neascus-type metacercariae, which cause the black spot disease, are described in specimens collected from 33 samples of Geophagus brasiliensis from the Peixe river, in Monte Verde, Juiz de Fora, state of Minas Gerais, Brazil, during March and May 2006. The main morphological features of these metacercariae are: genital primordia composed of three cell masses located in the hind body, and a median dorsal vessel of the reserve bladder around the acetabulum. Thirty fish specimens (90.9% were parasitized by metacercariae, with mean infection intensity of 15.6 ± 16.1 (1-75. Also, a positive correlation was detected between G. brasiliensis total length and the parasite abundance (r = 0.459; p < 0.01.
Egyptian Journal of Biology ... At the ventral side, the rims of the oral and ventral suckers had minor differences, while their floor showed great variation in topography: the floor of the oral sucker had longitudinal wrinkles with nodule-like structures and their upper margins had minute sensory papillae, while the ventral sucker ...
Metacercárias de Diplostomidae (Digenea: Diplostomoidea em Geophagus brasiliensis (Perciformes: Cichlidae do rio Guandu, Estado do Rio de Janeiro, Brasil - doi: 10.4025/actascibiolsci.v34i2.5957 Diplostomidae metacercariae (Digenea: Diplostomoidea in Geophagus brasiliensis (Perciformes: Cichlidae from Guandu river, State of Rio de Janeiro, Brazil - doi: 10.4025/actascibiolsci.v34i2.5957
José Luis Fernando Luque
Full Text Available O presente trabalho teve como objetivo o estudo qualitativo e quantitativo das metacercárias de diplostomídeos em Geophagus brasiliensis (Quoy & Gaimard, 1824 do rio Guandu, Seropédica, Rio de Janeiro, Brasil. Entre abril de 2006 e março de 2007, foram coletados e necropsiados 200 espécimes de G. brasiliensis, sendo obtidos cinco morfotipos de metacercárias de diplostomídeos: Austrodiplostomum compactum, Diplostomum sp., Posthodiplostomum sp. 1, Posthodiplostomum sp. 2 e Neascus tipo 1. No presente trabalho, foram descritas as metacercárias de Diplostomum sp., Posthodiplostomum sp. 1 e Posthodiplostomum sp. 2, além de registrar novo sítio de infecção de A. compactum e aumentar o conhecimento da distribuição geográfica de Neascus tipo 1, causadora da doença dos pontos pretos. O presente trabalho aponta para a importância de G. brasiliensis como hospedeiro intermediário de trematódeos diplostomídeos e sobre a diversidade dessas larvas no rio Guandu.The qualitative and quantitative analyses of diplostomid metacercariae in Geophagus brasiliensis (Quoy & Gaimard, 1824 of the Guandu river, Seropédica, Rio de Janeiro, Brazil, are provided. Between April 2006 and March 2007, 200 specimens of G. brasiliensis were collected and necropsied. Further, five morphotypes of diplostomid metacercariae were obtained, namely, Austrodiplostomum compactum, Diplostomum sp., Posthodiplostomum sp. 1, Posthodiplostomum sp. 2 and Neascus type 1. Whereas the metacercariae of Diplostomum sp., Posthodiplostomum sp. 1 and Posthodiplostomum sp. 2 were described, a new site of infection of A. compactum was registered and a new geographical report for Neascus type 1, responsible for the black-spot disease, has been added. Current investigation enhances the importance of G. brasiliensis as an intermediate host of diplostomid trematodes and the diversity of these larvae in the Guandu river.
Metacercárias de Diplostomum (Austrodiplostomum compactum Lutz, 1928 (Digenea, Diplostomidae em peixes do rio Paraná, Brasil. Prevalência, sazonalidade e intensidade de infecção Metacercariae of Diplostomum (Austrodiplostomum compactum Lutz, 1928 (Digenea, Diplostomatidae in fishes of Paraná River, Brazil. Prevalence, seasoning and intensity of infection
Haroldo Kazuyuki Takahashi
parasites (55 % with intensity rate of 9.3 parasites. Corvinas showed prevalence from 71 to 100 % with the highest intensity rates (103.3 and 106.9 from February to March, 2001. Comparatively, the tucunaré showed the highest prevalence from November 2000 (90% to February 2001 (80%. The tucunaré highest intensity rates occurred on September, 2000 (12.7, December, 2000 (12.2 and February, 2001 (16.1. Metacercariae showed a hollow womb body and a posterior region of a sinuous conic protuberance, an oral sucker smaller than the acetabulae, a simple acetabulae located in the pre-equatorial region of helminto. A short pre-pharynx followed by a muscular pharynx, genital pore next to acetubulae. Corvina metacercariae presents 1.434,0 µm (880 to 1.840 long by 611.2 ± 93.4 µm (400 to 792 wide. Tucunaré metacercariae, 1.462.4 µm (960 to 2.480 by 710.8 µm (560 to 960. Authors confirmed corvina great susceptibility to Diplomastum and the relationship between the parasitism and the aquatic and climatic parameters.
Botnevik, C.F.; Malagocka, Joanna; Jensen, Annette Bruun
. This behavior, hypothesized to involve cramping of the mandibular muscles in a state of tetany, was observed in naturally infected F. polyctena under controlled temperature, light, and humidity conditions. We found that low temperature significantly stimulated and maintained tetany in infected ants while light......, humidity, ant size, and infection intensity had no influence on this behavior. Under none of the experimental conditions did uninfected ants attach to vegetation, demonstrating that tetany was induced by D. dendriticum . Temperature likely has a direct impact on the initiation of clinging behavior...
Addino, Mariana; Lomovasky, Betina J.; Cremonte, Florencia; Iribarne, Oscar
Parasite life cycles are frequently completed in different hosts, thus the parasites have its life cycle overlapped to natural trophic webs. The family Gymnophallidae (Class: Trematoda; Subclass: Digenea) includes digenetic parasites whose larval stages occur on bivalves and may affect bivalve predation by the final host of these parasites. In this work we evaluated: (a) if individuals of the razor clam Tagelus plebeius with higher parasite intensity suffer higher predation by the oystercatcher Haematopus palliatus and, (b) if there is any effect of parasite intensity on burrowing and escape behaviours of these razor clams which may enhance exposure to predators. Field experiments (oystercatcher exclusion vs. open access) showed that clams with higher parasite intensity support higher predation by oystercatchers, which suggests a higher consumption of more parasitized clams and thus, a more successful reproduction of parasites linked to the intensity of infection. However, clam burrowing and escape behaviours did not show differences related to different parasite intensity, suggesting that the commonly believed mechanisms are not responsible in this case.
Kvach, Yuriy; Jurajda, Pavel; Bryjová, Anna; Trichkova, T.; Ribeiro, F.; Přikrylová, I.; Ondračková, Markéta
Roč. 66, č. 5 (2017), s. 635-642 ISSN 1383-5769 R&D Projects: GA ČR(CZ) GBP505/12/G112 Institutional support: RVO:68081766 Keywords : Lepomis gibbosus * Micropterus salmoides * White grub * Physid snails * Non-indigenous species Subject RIV: EG - Zoology OBOR OECD: Parasitology Impact factor: 1.744, year: 2016
Ondračková, Markéta; Reichard, Martin; Jurajda, Pavel; Gelnar, M.
Roč. 93, č. 2 (2004), s. 131-136 ISSN 0932-0113 R&D Projects: GA ČR GA524/02/0924 Institutional research plan: CEZ:AV0Z6093917 Keywords : black-spot disease * juvenile fish Subject RIV: EG - Zoology Impact factor: 1.060, year: 2004
Yang, Bin-Bin; Zhou, Ben-Jiang; Li, Ru-Qing; Bai, Zhong-Wen; Wu, Ou-Bao; Gao, Xiu-Fang
69 crabs were collected from Daxing, Gekui and Niukong townships of Lvchun county, Yunnan Province in 2006 and excysted metacercariae were only obtained from crabs of Niukong. The infection rate was 27.6% (8/29) with an average metacercaria number of 2.25 each crab. No encysted metacercariae were found. The excysted metacercariae were morphologically identified as Paragonimus proliferus.
Ondračková, Markéta; Bartošová, Šárka; Valová, Zdenka; Jurajda, Pavel; Gelnar, M.
Roč. 49, č. 3 (2004), s. 222-227 ISSN 1230-2821 R&D Projects: GA ČR GA524/02/0924 Institutional research plan: CEZ:AV0Z6093917 Keywords : Digenea * juvenile fish * host susceptibility Subject RIV: EG - Zoology Impact factor: 0.560, year: 2004 http://www.actaparasitologica.pan.pl/archive/49/toc49130.html
Gómez del Prado Rosas, Maria del Carmen; Valles Vega, Isabel; Lozano Cabo, Horacio; Hernández Trujillo, Sergio
En la Bahía y Ensenada de La Paz existe una fuerte variabilidad en la composición y abundancia de especies de copépodos pelágicos, pero no existe información de los niveles de infección por parásitos. Las primeras fases del ciclo de vida de los tremátodos es poco claro, debido a que utilizan medusas, ctenóforos, moluscos, quetognatos, copépodos y larvas de peces como primeros y segundos hospederos intermediarios. Por lo que el objetivo de este trabajo fue el de conocer los helmintos que prese...
Skov, Jakob; Kania, Per Walter; Jørgensen, Thomas Rohde
Den digene ikte Pseudamphistomum forekommer som metacercarie i skaller fra Furesøen og som adult i mink fra området. Dette er belyst ved morfometriske og molekylære metoder. Fundet er væsentligt, idet parasitten er zoonotisk og kan inficere mennesker, der indtager utilstrækkeligt varmebehandlet f...
from Aguaytia, an undescribed metacercaria of Paragonimus was found from one of 13 crabs, Pseudothelphusa chilensis collected in the university campus...be infected with Paragonimus metacercariae, which were separated into two species by their characteristics. The first species was identified as P.peruvianus, and the second one was presumed to belong to P.mexicanus. (Author)
Renato Ribeiro Nogueira Ferraz
Full Text Available The demand and consumption of fish and their derivatives has increased considerably in recent years. However, fish are ideal hosts of numerous parasites, highlighting the need to develop new research methodologies for its detection. The aim of present study was to compare the efficacy of Ascocotyle metacercariae (Trematoda: Digenea extraction from visceral tissues of Mugil liza Valenciennes, 1836 (Teleostei: Mugilidae by two methods: homogenization by blender or mixer. Twentysix samples of M. liza were collected, being 16 liver samples and 10 samples of muscle tissue. Approximately 5g of each sample were processed by blender and mixer techniques homogenization for metacercariae extraction. In liver samples, up to 46 metacercariae were found in samples homogenized in blender. The lowest amount found was 2 metacercariae for blender and mixer techniques. In samples of muscle tissue, 4 metacercariae were observed in the mixer extraction. The lowest amount was found to be 2 parasites to blender and mixer. The mean metacercariae found and extracted from muscle tissue were 0.2 (+0.357 and 1.2 (+0.963 for blender and mixer, respectively. The averages of metacercariae found and extracted from fish liver, in blender and mixer, were 24 (+15.145 and 18 (+8.246, respectively. The homogenization techniques for blender and mixer were effective for the extraction of metacercariae of mullet fish tissues, suggesting that they may be directly applicable in the field of study, especially due to the ease of testing.
The liver pathology of Clarias gariepinus infected with Euclinostomum clarias metacercariae in Anambra river basin was investigated from January to September, 2002. The clariids were examined and the livers of those infected with the metacercariae of the digenean trematode, Euclinostomum clarias, were fixed in 10% ...
Liu, Jian-Fa; Wu, Yan; Tang, Zhi-Yuan; Meng, Dan; Yang, Shu-Juan; Jiang, Wen-Wen
Freshwater crabs and snails were collected from Ninghai County in Zhejiang Province, and examined respectively for Paragonimus metacercariae and cercariae. Among 97 freshwater crabs found, the prevalence was 11.3% (11/97) with a mean intensity of 1 metacercariae per crab. It was 10.2% (5/49) and 20.2% (4/20) in the groups weighted 5-15 g and 15-25 g respectively, with an average intensity of 1, and no metacercariae were found in weight group of 25-35 g. Two positive crabs were found from 20 crabs with a low weight (< 5 g). Male to female crabs ratio was 2.5:1, and there was no significant difference in prevalence between males [12.7%(7/55)] and females [9.1% (2/22)]. No cercariae or metacercariae were found in 200 snails (Semisulcospira libertino).
Description and proposed life cycle of Maritrema novaezealandensis n. sp. (Microphallidae) parasitic in red-billed gulls, Larus novaehollandiae scopulinus, from Otago Harbor, South Island, New Zealand
Martorelli, Sergio R; Fredensborg, Brian Lund; Mouritsen, Kim Nørgaard
Maritrema novaezealandensis n. sp. is described from Otago Harbor, South Island, New Zealand, on the basis of adult specimens collected from the Red-billed gull, Larus novaehollandiae scopulinus, and excysted metacercariae obtained from crabs. It belongs to the "eroliae group" and differs from......, with metacercariae encysted in the body cavity of the crustacean host. Finally, the definitive host, the gull, L. n. scopulinus, harbors the adult worms in its intestine....
In Tingo Maria, Peru, 25 crabs, Pseudothelphusa chilensis were examined for Paragonimus metacercariae with negative results. Five lung flukes were...minimus. These flukes belonged to the same species characterized by their extremely elongated body, and were named Paragonimus amazonicus sp. n. by...harbored 44 Paragonimus metacercariae in all. These larvae were divided into two species, five of them being identified as Paragonimus peruvianus and
Pinto, H A; Melo, A L
Melanoides tuberculata , naturally infected by gymnocephalous cercariae, were found in aquatic collections from Belo Horizonte, Minas Gerais, Brazil. After morphological characterization, larvae were used for experimental infection of Poecilia reticulata. Metacercariae were obtained from the liver of these fish, which were also found to be naturally infected in the same locality. The morphology and biology of the developmental stages of trematodes we obtained were characteristic of Renicola sp. This is the first record of renicolid cercariae and metacercariae in Brazil.
Shaw, J.C.; Hechinger, R.F.; Lafferty, Kevin D.; Kuris, Armand M.
We describe the distribution and abundance of the brain-encysting trematode Euhaplorchis californiensis and its second intermediate host, the California killifish (Fundulus parvipinnis), in 3 estuaries in southern California and Baja California. We quantified the density of fish and metacercariae at 13–14 sites per estuary and dissected 375 killifish. Density (numbers and biomass) was examined at 3 spatial scales, i.e., small replicate sites, habitats, and entire estuaries. At those same scales, factors that might influence metacercaria prevalence, abundance, and aggregation in host individuals and populations were also examined. Metacercaria prevalence was 94–100% among the estuaries. Most fish were infected with 100s to 1,000s of E. californiensis metacercariae, with mean abundance generally increasing with host size. Although body condition of fish did not vary among sites or estuaries, the abundance of metacercariae varied significantly among sites, habitats, estuaries, and substantially with host size and gender. Metacercariae were modestly aggregated in killifish (k > 1), with aggregation decreasing in larger hosts. Across the 3 estuaries, the total populations of killifish ranged from 9,000–12,000 individuals/ha and from 7–43 kg/ha. The component populations of E. californiensis metacercariae ranged from 78–200 million individuals/ha and from 0.1–0.3 kg/ha. Biomass of E. californiensis metacercariae constituted 0.5–1.7% of the killifish biomass in the estuaries. Our findings, in conjunction with previously documented effects of E. californiensis, suggest a strong influence of this parasite on the size, distribution, biomass, and abundance of its killifish host.
Timi, J T; Martorelli, S R; Sardella, N H
A total of 2,086 specimens of Engraulis anchoita Hubbs et Marini (Pisces: Engraulidae), from waters off Argentina and Uruguay were examined for parasitic digeneans. Four species of digeneans were found, Parahemiurus merus (Linton, 1910) (adult) in the stomach; Lecithochirium microstomum Chandler, 1935 (non-ovigerous and ovigerous juveniles) in the swimbladder; Cardiocephaloides sp. (metacercaria) in the eyes and Rhipidocotyle sp. (metacercaria) in the musculature. These four species are recorded for the first time as parasites of E. anchoita.
Doanh, Pham Ngoc; Shinohara, Akio; Horii, Yoichiro; Habe, Shigehisa; Nawa, Yukifumi; Le, Nguyen Thi
Based on morphology of metacercariae and adult worms together with molecular data from our previous study, we describe herein a new lung fluke species named Paragonimus vietnamensis sp. nov. Metacercariae of this new species is round and extremely large in size (nearly 800 mum in diameter) having a fragile outer cyst of variable thickness and a thin fragile inner cyst. There are little or no space between metacercaria and the inner cyst wall. These and other morphological features of metacercariae of P. vietnamensis sp. nov. are not completely identical with but have some similarities with those of P. microrchis, P. proliferus, or P. menglaensis. On the other hand, adult worms obtained by experimental infection of these large metacercariae are oval in shape, having a ventral sucker slightly larger than the oral one, and having singly arranged relatively short cuticular spines. These morphological features of adult worms are partly similar to but not identical with those of P. microrchis, P. skrjabini, P. yunnanenis, P. xiangshanensis, or P. harinasutai. Taken these morphological data and our previous molecular phylogenetic analyses of ITS2 and CO1 sequences of metacercariae and adult worms together, P. vietnamensis sp. nov. is different from any other known Paragonimus spp.
Gymnophalloides seoi is a peculiar human intestinal trematode in Korea transmitted by oysters. This study was carried out to observe the effects of radiation on the infectivity of G. seoi metacercariae to C3H mice and to assess the applicability of radiation for use in the control of gymnophalloidiasis. Oysters were collected from the endemic area. Non-irradiated control, metacercaria-irradiation, and oyster-irradiation groups were prepared. One hundred metacercariae were infected orally to each mouse, and worm recovery rates of three groups were compared at the seventh day post-infection. In the metacercaria-irradiation group, the worm recovery rate was significantly reduced at radiation doses higher than 200 Gy, and the number of intrauterine eggs was significantly reduced at doses over 50 Gy. In the oyster-irradiation group, 50 Gy significantly reduced the worm recovery rate and number of uterine eggs. In the two groups, no worm was recovered at 1,000 Gy. In conclusion, G. seoi metacercariae showed some resistance to radiation at lower doses than 200 Gy, but irradiation of oyster with 200-1,000 Gy could be applied as a control measure for gymnophalloidiasis. (author). 10 refs, 3 figs, 2 tabs
Doanh, P N; Thaenkham, U; An, P T; Hien, H V; Horii, Y; Nawa, Y
Paragonimus heterotremus, which is an important pathogen for human paragonimiasis in Asia, is recognized as having the smallest metacercariae (maximum diameter Paragonimus species. Recently, P. pseudoheterotremus has been described from Thailand as a new species having metacercariae (about 200 μm) slightly smaller than those of Thai P. heterotremus. In fact, the small size of P. pseudoheterotremus metacercariae is compatible with those of P. heterotremus from India and China. In this study in Vietnam, we found variably sized small metacercariae which are expected to consist of both P. heterotremus and P. pseudoheterotremus. Contrary to expectation, the adult flukes obtained by separate infection of experimental cats with different sized metacercariae were all identified as P. heterotremus, using both morphological and molecular characteristics. The molecular analyses of an extensive collection of P. heterotremus/P. pseudoheterotremus isolates from Asian countries also indicated that genetic distances between different populations of P. heterotremus are even larger than that between P. pseudoheterotremus and P. heterotremus. The haplotype network showed that all P. heterotremus and P. pseudoheterotremus isolates formed a P. heterotremus complex consisting of three groups with strong geographical origins. In addition, the Indian P. heterotremus group is the root of the other P. heterotremus and P. pseudoheterotremus populations. Based on the observed metacercarial polymorphisms and genetic variation in P. heterotremus, P. pseudoheterotremus should be considered a geographically isolated population of the P. heterotremus complex.
Doanh, Pham Ngoc; Shinohara, Akio; Horii, Yoichiro; Habe, Shigehisa; Nawa, Yukifumi; Le, Nguyen Thi
During an epidemiological survey for Paragonimus and paragonimosis in northern Vietnam, we found extremely large excysted metacercariae (2.50 +/- 0.14 mm in length and 0.72 +/- 0.08 mm in width; mean +/- standard deviation of 20 samples) in mountainous crabs, Potamiscus mieni. Adult worms were successfully obtained by intraperitoneal injection with those large excysted metacercariae in a cat. Morphological and morphometric data of those large excysted metacercariae and the adult worms derived from them are identical to those of Paragonimus proliferus found in Yunnan province, China. However, when second internal transcribed spacer region and cytochrome oxidase subunit 1 gene sequences of those metacercariae and adult worms were compared with those of known Paragonimus spp. deposited in the GenBank, they were almost completely identical to those of Paragonimus hokuoensis metacercariae in China, of which adult worms have never been reported. This is the first record of P. proliferus in Vietnam and the first record from outside of China. Phylogenetic relationship between P. proliferus and P. hokuoensis is discussed.
Torres, J.L.; Chiriboga, J.
Fascioliasis or hepatic distomatosis is one of the most widespread liver diseases throughout the world. It is caused by a parasitic worm of the class Trematoda of the phylum Platyhelminthes. This flatworm is commonly known as liver-fluke in the United States and Europe; babosa del higado in Cuba; cucaracha del higado in Puerto Rico; saguaype in Argentina and Chile; pirihuin in Chile; yuta in North Chile, and it is taxonomically classified as Fasciola hepatica. The adult trematode is a common parasite of warm blooded animals especially cattle, sheep, and goats. Alterations in the viability of normal Fasciola hepatica metacercariae as a result of time of storage at 4/sup 0/C were studied by the method of Wikerhauser (1960). A significant decrease in viability was observed only after one hundred days of storage. The in vitro viability of normal metacercariae was compared with that of metacercariae that were exposed to increasing ..gamma.. radiation doses. Two plainly opposite effects observed were: the activation of the excystation process by the lower radiation doses (1.5 and 2.5 Kr), an effect that persisted up to the 14th day post-radiation, and the impairment of the process evidenced on the 14th day in the metacercariae exposed to 3.5 and 5.0 Kr. Correlation between liver damage and the increasing radiation doses to which the metacercariae were exposed was studied up to 34 days after the oral inoculation in rats. (auth)
Torres, J.L.; Chiriboga, J.
Fascioliasis or hepatic distomatosis is one of the most widespread liver diseases throughout the world. It is caused by a parasitic worm of the class Trematoda of the phylum Platyhelminthes. This flatworm is commonly known as liver-fluke in the United States and Europe; babosa del higado in Cuba; cucaracha del higado in Puerto Rico; saguaype in Argentina and Chile; pirihuin in Chile; yuta in North Chile, and it is taxonomically classified as Fasciola hepatica. The adult trematode is a common parasite of warm blooded animals especially cattle, sheep, and goats. Alterations in the viability of normal Fasciola hepatica metacercariae as a result of time of storage at 4 0 C were studied by the method of Wikerhauser (1960). A significant decrease in viability was observed only after one hundred days of storage. The in vitro viability of normal metacercariae was compared with that of metacercariae that were exposed to increasing γ radiation doses. Two plainly opposite effects observed were: the activation of the excystation process by the lower radiation doses (1.5 and 2.5 Kr), an effect that persisted up to the 14th day post-radiation, and the impairment of the process evidenced on the 14th day in the metacercariae exposed to 3.5 and 5.0 Kr. Correlation between liver damage and the increasing radiation doses to which the metacercariae were exposed was studied up to 34 days after the oral inoculation in rats
Marcos T. Díaz
Full Text Available Descriptions of the miracidium, mother redia, daughter redia, grandaughter redia, cercaria, metacercaria and adult stages of Philophthalmus gralli Mathis and Leger,1910 recovered from experimental infections are presented. The intermediate host, Melanoides tuberculata, was collected from freshwater rivulets in Aguasanta and Yaguracual, Sucre State, Venezuela. Chicken were orally infected with cercariae and metacercariae, and metacercariae were introduced directly into the eyes by pipette. Both processes of infection produced adult worms. This is a new geographical record for P. gralliSe describe el miracidio, redia madre, redia hija, redia nieta, cercaria y el adulto de Philophthalmus gralli Mathis y Leger, 1910, obtenidos de infecciones experimentales. El hospedero intermediario Melanoides tuberculata fue recolectado en las localidades de Aguasanta y Yaguaracual, estado Sucre,Venezuela. Varios pollos fueron infectados oralmente con cercarias y metacercarias y colocando metacercarias directamente en los ojos con una pipeta. En ambos procesos de infección, se obtuvo parásitos adultos. Se amplía la distribución geográfica de P. gralli
Rekha Devi, K; Narain, Kanwar; Mahanta, Jagadish; Nirmolia, Tulika; Blair, David; Saikia, Sidhartha P; Agatsuma, Takeshi
The name Paragonimus westermani (Kerbert, 1878) is commonly applied to members of a species complex that includes the well-known Asian lung fluke of medical and veterinary importance. Unambiguous molecular and morphological evidence showing the presence of a member of the complex in India has recently been published. In the present study we report the occurrence of 2 more members of the P. westermani complex in northeastern (NE) India. Surveys of the freshwater crabs Maydelliatelphusa lugubris in NE India revealed 2 morphologically distinct types of lung fluke metacercariae. Phylogenetic analyses, using DNA sequences from ITS2, 28S and cox1 gene regions indicate that these lung metacercariae belong to P. westermani complex. Type 1 metacercariae have a more basal position within the complex whereas type 2 metacercariae are closely related to the relatively derived forms of P. westermani from NE Asia (Japan, Korea, China) and Vietnam. A third type of metacercaria (type 3), detected in another crab host, Sartoriana spinigera in Assam, was phylogenetically close to P. siamensis, also a member of the P. westermani group. Molecular evidence has demonstrated the existence of 3 genotypes of lung flukes within the Paragonimus westermani complex in NE India. Two of these were previously unknown.
Sohn, Woon-Mok; Ryu, Jae-Sook; Min, Duk-Young; Song, Hyun-Ouk; Rim, Han-Jong; Vonghachack, Youthanavanh; Bouakhasith, Daluny; Banouvong, Virasack
Paragonimus harinasutai metacercariae were found in a species of freshwater crab, Indochinamon ou, collected in a small stream of Namback District, Luang Prabang Province, Lao PDR. Adult flukes were recovered after experimental infection of the metacercariae to dogs. Metacercariae were round or slightly elliptical, 0.666 x 0.626 mm in average size, and had a thin cyst wall of about 20 microm in thickness, a black excretory bladder, convoluted ceca, and some pinkish materials in the body. Adults were somewhat elongated, 95.2 x 36.5 mm in average size, covered with single-tipped tegumental spines, had a smaller oral sucker than the ventral sucker, a moderately branched ovary, and 5-6 lobulated testes. Eggs were ovoid and bilaterally symmetrical in shape, 79 x 45 microm in average size, and had a uniformly thickened shell. By the present study, it has been confirmed that I. ou is a new second intermediate host for P. harinasutai.
Petrov, A A; Podvyaznaya, I M
Metacercarial development of most Diplostomum species including D. pseudospathaceum occurs in the eye lenses of their fish hosts and is accompanied by radical morphological changes often referred to as metamorphosis. One of the structures undergoing substantial changes in D. pseudospathacewn are tegumental spines. The present study used phalloidin staining and confocal microscopy to examine these changes in D. pseudospathaceum over the course of development from 3-day-old to infective metacercariae. Although the general pattern of spination remained essentially unchanged, most larval spines continued growing in size until late in metacercarial development. From day 10 of development, larval spination was gradually replaced by small incipient definitive spines and the infective metacercariae had only definitive spination. The possible adaptive role of spines in developing metacercariae is discussed.
Guk, Sang-Mee; Chai, Jong-Yil; Sohn, Woon-Mok; Kim, You-Me; Sim, Seobo
Microphallus species occur primarily as intestinal parasites of birds and mammals, and metacercariae of a new species belonging to this genus have been discovered from the crab, Macrophthalmus dilatatus, in the Republic of Korea. The metacercaria of this fluke was round with 2 thick walls, and the excysted one had mature genital organs. The adult flukes recovered from experimentally infected chicks had numerous intrauterine eggs, well-developed pars prostatica, widely bifurcating ceca, and prominent uterine bulge. After observing internal structures, it was concluded that this species is different from any other known Microphallus spp. Based on the morphology of metacercariae and adult flukes, we describe this specimen as a new species, Microphallus koreana n. sp. PMID:18830056
Suanyuk, Naraid; Mankhakhet, Suchanya; Soliman, Hatem; Saleh, Mona; El-Matbouli, Mansour
During April to June 2009 and February 2010 to February 2011, numerous digenetic trematode metacercariae were observed embedded in the muscles of guppies Poecilia reticulata cultured at Kidchakan Supamattaya Aquatic Animal Health Research Center, Songkhla, southern Thailand. A total of 424 guppies were examined to identify the parasite and to study its pathogenicity. Based on conventional parasitological techniques, the parasitic digenean found in the guppies was identified as Euclinostomum heterostomum Rudolphi, 1809. Histological analysis revealed numerous metacercariae embedded in the fish muscle. A life cycle study indicated that the snail Indoplanorbis exustus was the first intermediate host, with the guppies serving as the second intermediate host. No E. heterostomum metacercariae were found in cohabited fish species, giant sailfin molly Poecilia velifera or platy Xiphophorus maculatus, which indicated that the guppy was the only suitable fish host present.
Bui, Thanh Ngoc; Pham, Thanh Thi; Nguyen, Nguyen Thi; Nguyen, Ha Van; Murrell, Darwin; Phan, Van Thi
Preliminary findings of a high prevalence of Clonorchis sinensis in wild-caught fish in a North Vietnam reservoir (Thac Ba reservoir, Yen Bai Province) prompted a longitudinal epidemiological study of fish infections. Monthly collections of fish from September 2014 to August 2015 were processed for recovery of metacercariae; 1219 fish, representing 22 species, were examined. Seven species were infected with C. sinensis metacercariae. Four species, Toxabramis houdemeri, Hemiculter leucisculus, Cultrichthys erythropterus, and Culter recurvirostris, had high prevalence (31.1 to 76.7 %); metacercarial intensities ranged from 3.9 to 65.7 metacercariae/fish. A seasonal variation of C. sinensis prevalence was observed in T. houdemeri. Variation in intensity of infection occurred in C. erythropterus and H. leucisculus. Intensity and prevalence of C. sinensis in the most highly infected species, T. houdemeri, varied by fish size; prevalence was higher in fish weighing more than 3 g, and intensity was higher in fish weighing more than 5 g. The distribution of metacercariae in the body region of T. houdemeri was significantly higher in the caudal fin (14.7 metacercariae/g), compared to the body and head regions (0.7 and 1.4 metacercariae/g, respectively). Further epidemiological investigations on C. sinensis in this reservoir region should include assessing the relative risk of the different fish species for humans based on the latter's food preferences, and the prevalence of C. sinensis in the community. The snail intermediate host(s) in the reservoir should also be identified along with the ecological factors influencing its exposure to C. sinensis eggs and its subsequent transmission of cercariae to fish. Also needed are investigations on the relative importance of wild and domestic reservoir hosts as sources of egg contamination of the reservoir.
Angelika Anna Linowska
Full Text Available Paracoenogonimus ovatus is the parasite that infest wide definitive host range (predator birds and at least two intermediate hosts: molluscs (Viviparus viviparus, V. fasciatus and freshwater fish from eastern and central Europe. The larvae of the parasite (metacercariae go to the muscle and connective tissue, and at high intensity of infection to the internal organs and brain of fish. A total of 128 fishes were caught in autumn 2009 and 2014, by fishing boats operating in Lake Dąbie, which is one of the largest lakes in Poland. The species selected reflect the species structure of commercial and recreational catches. They were roach, bream, silver bream, crucian carp (Cyprinidae, ruffe, perch, zander (Percidae and pike (Esocidae. The fish were filleted, and the skinned muscle was compressed under a trichinoscope, what allowed to see and count metacercariae in the tissues. To facilitate parasite isolation from the cysts, the muscle tissues were subjected to a 0.1 % solution of activated pepsin and 5 % citric acid at a temperature of 20ºC. The free larvae were measured and identified under an Olympus BX 50 microscope coupled with a camera running AxioVs40 V 18.104.22.168 software. Metacercariae are grouped primarily at depths ranging from just beneath the skin to about 50% of fillet thickness. The infection prevalence of P. ovatus metacercariae in the fish analyzed was 78,5%. The density of infection per gram of muscle tissue mass in the fish infected was 8.85 (from 1 to 84 metacercariae. The highest density was noted in roach, bream and white bream (17.09, 11.14 and 9.58, respectively. Metacercariae occurred singly in pike, parasites were not found in the muscles of crucian carp, ruffe, and perch examined. The reason for such differences in infection is the availability and composition of food, as well as the behavior of the fish examined.
Mehrdana, Foojan; Jensen, Hannah M.; Kania, Per Walter
Ornamental fish, Xiphophorus maculatus, were imported from Singapore to Denmark for distribution to local aquarists. Importers observed lethargic and erratic swimming patterns among fish and forwarded a total of 30 fish for pathological examination to a university diagnostic service. All fish were...... diagnosed infected with encysted Centrocestus sp. metacercariae in gills (prevalence of 100% and mean intensity of 454.5 ± 161.9 parasites per fish). Metacercariae were identified by morphological and molecular methods. Cysts (mean length 163.3 ± 13.7 μm and mean width 113.3 ± 10.6 μm) contained a bent...
Sokolov, S G; Frenkel, S E; Gordeev, I I
Samples of Zooplankton collected in waters of the Prostor Gulf (Iturup Island) were examined. Metacercariae of Brachyphallus crenatus were found in copepods Pseudocalanus newmani and Acartia longiremis. This is the first record of the second intermediate hosts of this species in the North Pacific.
Sokolov, S G; Gordeev, I I
Metacercariae of Crepidostomum metoecus (prevalence 2.0%), procercoids of Cyathocephalus truncatus (2.9%), and nematodes of the family Cystidicolidae (17.6%) were found in the body cavity of amphipods Gammarus lacustris from the Kronotskoe Lake. The nematodes were presented by Cystidicola farionis (third-stage juvenile) and Ascarophis sp. (adult and fourth-stage juvenile).
Wang, Rui-feng; Tang, Ling-jing; Zhou, Ai-ming
Four adults of Fasciola hepatica were found from the bile ducts of a patient diagnosed as biliary calculi during a surgical operation. We investigated retrospectively the infection source and concluded that the patient may be infected by eating raw or half-cooked Zizania latifolia, an aquatic plant, which was contaminated with metacercariae of F. hepatica.
de Núñez, Margarita Ostrowski
Austrodiplostomum compactum from Nannopterum brasilianus, and its metacercaria from Geophagus sp. and Oreochromis mossambicus captured (1979) at its type locality, Valencia Lake, Venezuela, by the author, are redescribed. The adult is characterized by its large body size, and an oral sucker smaller than the pharynx. The metacercaria has a similar body size as the adult, and the small genital primordia occupy 4.1-7.3% of body length. Experimental infections in chickens with metacercariae of Diplostomulum mordax from brains of Odonthestes bonariensis, captured (2015) at Dique Paso de las Piedras, near Bahia Blanca City, Buenos Aires Province, Argentina, resulted in 10 adults 5 days postexposure. These adults correspond to Austrodiplostomum mordax as described from N. brasilianus at Lacombe Lagoon, Buenos Aires Province, and differ from A. compactum in their smaller body size, and an oral sucker larger than the pharynx. The metacercaria has a similar body size as the adult and differs mainly in that the larger genital primordia occupy 11.6-13.8% of body length. The status of earlier published Austrodiplostomum species in the American continent is discussed in view of available morphological and molecular data. A lectotype of A. mordax is here designated, and Austrodiplostomum ostrowskiae is considered as a new synonym of A. compactum.
Kondo, Yusuke; Ohtsuka, Susumu; Hirabayashi, Takeshi; Okada, Shoma; Ogawa, Nanako O; Ohkouchi, Naohiko; Shimazu, Takeshi; Nishikawa, Jun
In the Seto Inland Sea of western Japan, metacercariae of three species of trematodes, Lepotrema clavatum Ozaki, 1932, Cephalolepidapedon saba Yamaguti, 1970, and Opechona olssoni (Yamaguti, 1934), were found in the mesoglea of the jellyfish Aurelia aurita s.l., Chrysaora pacifica, and Cyanea nozakii. Moreover, these jellyfish frequently harbored juveniles of the fish species Psenopsis anomala, Thamnaconus modestus, and Trachurus japonicus. The former two fish species are well-known medusivores. We investigated seasonal changes in the prevalence and intensity of these metacercariae in their host jellyfish from March 2010 to September 2012 and presumed that infection by the trematodes of the definitive host fish occurs through these associations. The mean intensity of metacercariae in A. aurita s.l. clearly showed seasonality, being consistently high in June of each year. The intensity of metacercariae in C. nozakii was highest among all jellyfish hosts and appeared to be enhanced by medusivory of this second intermediate, and/or paratenic host. Trophic interactions between jellyfish and associated fish were verified using both gut content and stable isotope analyses. The detection of trematodes and nematocysts in the guts of P. anomala and T. modestus juveniles, in addition to stable isotope analysis, suggests that transmission of the parasites occurs via prey-predator relationships. In addition, the stable isotope analysis also suggested that P. anomala is more nutritionally dependent on jellyfish than Th. modestus and Tr. japonicus. © Y. Kondo et al., published by EDP Sciences, 2016.
Full Text Available In the Seto Inland Sea of western Japan, metacercariae of three species of trematodes, Lepotrema clavatum Ozaki, 1932, Cephalolepidapedon saba Yamaguti, 1970, and Opechona olssoni (Yamaguti, 1934, were found in the mesoglea of the jellyfish Aurelia aurita s.l., Chrysaora pacifica, and Cyanea nozakii. Moreover, these jellyfish frequently harbored juveniles of the fish species Psenopsis anomala, Thamnaconus modestus, and Trachurus japonicus. The former two fish species are well-known medusivores. We investigated seasonal changes in the prevalence and intensity of these metacercariae in their host jellyfish from March 2010 to September 2012 and presumed that infection by the trematodes of the definitive host fish occurs through these associations. The mean intensity of metacercariae in A. aurita s.l. clearly showed seasonality, being consistently high in June of each year. The intensity of metacercariae in C. nozakii was highest among all jellyfish hosts and appeared to be enhanced by medusivory of this second intermediate, and/or paratenic host. Trophic interactions between jellyfish and associated fish were verified using both gut content and stable isotope analyses. The detection of trematodes and nematocysts in the guts of P. anomala and T. modestus juveniles, in addition to stable isotope analysis, suggests that transmission of the parasites occurs via prey-predator relationships. In addition, the stable isotope analysis also suggested that P. anomala is more nutritionally dependent on jellyfish than Th. modestus and Tr. japonicus.
To identify the species of Paragonimus proliferus with scanning electron microscopy (SEM) based on the surface structure of excysted metacercariae, adult worms and eggs. Crabs were collected from the endemic area of P. proliferus and excysted metacercariae were separated. Adult worms at different ages and eggs were obtained from the experimentally infected rats. After being fixed by 2.5% glutardialdehyde and 1% osmic acid, alcohol dehydration, gilded by ion spatter, the specimens were observed under SEM by STEREOSCAN-100. The cuticular spines of excysted metacercariae distributed in single pattern, bayonet-shaped or scale-shaped. There were 6 dome-shape papillae around the rim of the ventral sucker symmetrically arranged. The cuticular spines of different age adult worms distributed in group pattern, relatively denser and more regularly arranged in the anterior part than the posterior part of the worm body. The shape and arrangement of the cuticular spines on adult worms at different ages were basically uniform. The surface of eggshell including the operculum was generally smooth. The shell rim joining the operculum was thick and prominent. A knot-like prominence was observed at the aboperculum end. The cuticular spines of both excysted metacercariae and adult worms of P. proliferus show its own characteristics, but the size and shape of the cuticular spines among individuals or different parts of the same specimen show certain differences.
Full Text Available Experimental infections of Galba truncatula (two populations with Fasciola hepatica or Paramphistomum daubneyi were carried out to study the effect of water temperature changes (3 h at a mean of 12 °C every week on cercarial shedding during the patent period. The results were compared with those of control snails infected according to the same protocol and always maintained at 20 °C. Compared to controls, a significant increase in the number of cercariae-shedding snails, a significantly longer patent period and significantly greater cercarial production were noted in temperature-challenged snails, regardless of the type of digenean infection. In contrast, the number of incompletely formed metacercariae was significantly higher in temperature-challenged snails than in controls. Incompletely formed metacercariae of F. hepatica consisted of cysts whose colour remained whitish after shedding (25.4% for temperature-challenged snails or whose dome was flattened after encystment (74.6%. Those of P. daubneyi were totally dark brown or blackish after formation. These incomplete metacercariae might originate from young differentiating cercariae within the snail body (F. hepatica or from cercariae which died just after encystment (P. daubneyi. The use of regular temperature changes for snails infected with F. hepatica or P. daubneyi must be monitored carefully during collection of metacercariae to select completely formed cysts for infecting definitive hosts.
May 3, 1988 ... Unencysted trematode metacercariae of the genus Proctoeces were found embedded in the mantle tissue and visceral mass of the intertidal mussel Perna perna. Data on the prevalence and intensity of ..... temperature dependent, however, this seems unlikely in view of the known worldwide distribution of ...
So, freshwater crabs were demonstrated as one of the main hosts for P. metacercariae in Jinhua. Moreover, phylogenetic analysis indicated that the sequences of the second internal transcribed spacer (ITS2) and cytochrome coxidase subunit I (COI) were conserved among different isolates, especially for the ITS2 gene.
ABSTRACT: A study was made on some aspects of the biology of lymmzca truncatula and production of metacercariae of Fasciola hepatica under laboratory conditions. The e>'
Michálková, Veronika; Ondračková, Markéta
Roč. 84, č. 5 (2014), s. 1377-1388 ISSN 0022-1112 R&D Projects: GA ČR GBP505/12/G112 Institutional support: RVO:68081766 Keywords : Bitterling * Diplostomum pseudospathaceum * Metacercariae * Parasite infection * Trematoda Subject RIV: EG - Zoology Impact factor: 1.658, year: 2014
In this study, the rabbit was assessed as a laboratory host for the helminthes parasite, Fasciola gigantica. Three groups of rabbits were Infected experimentally with 5, 10 end 15 metacercariae of F. gigantica respectively. Clinical signs found included pale mucous membrane, progressive emaciation and rough hair coat.
Boerlage, A.S.; Graat, E.A.M.; Verreth, J.A.J.; Jong, de M.C.M.
Fish-borne trematodes are reported to affect the health of more than 40 million people worldwide. Few experimental studies are available on fish size dependent gain (attack rates of cercariae) or loss (mortality of metacercariae) of fish-borne trematodes. Aim was to quantify the relation between
Full Text Available A survey of cercariae and metacercariae (Trematoda, Digenea from the great pond snail (Lymnaea stagnalis in Central Europe (Austria, Czech Republic, South-East Germany, Poland and Slovak Republic is presented, based on a study of 3,628 snails examined from 1998 to 2005. A total of 953 (26.3% L. stagnalis were infected with 24 trematode species comprising 19 species of cercariae and 11 species of metacercariae (six species occurred both as cercarie and metacercarie of eight families. The dominant cercariae were those of Opisthioglyphe ranae (159 hosts infected, Plagiorchis elegans (141 (both family Plagiorchiidae and Echinoparyphium aconiatum (153 (Echinostomatidae; 14 double infections were found. The most frequent metacercariae were those of Neoglyphe locellus (71 (Omphalometridae, E. aconiatum (66, Echinostoma sp. (59 and Moliniella anceps (48 (Echinostomatidae. In the previous studies carried out in Central Europe, a very similar spectrum of nine trematode families of 22 cercariae determined to species level and 43 types of cercariae reported under generic or provisional names, which can be in many cases conspecific with the previous taxa, were found. A simple key to identification of cercariae and metacercariae, together with their illustrations, is provided.
This report concerns a collection of parasitic worms from South Africa, and contains a new species of monogenean, Gyrodactylus transvaalensis, six species of adult trematodes, including a new species of Phyllodistomum.P. van der waali, two metacercariae of the Family Clinostomidae and three of the Order Strigeida, ...
Haroun, E.M.; Yagi, A.I.; Younis, S.A.; El Sanhouri, A.A.; Gadir, H.A.; Gameel, A.A.; Bushara, H.O.
A primary infection with Fasciola gigantica, which had been eliminated by anthelmintic treatment after eight weeks, stimulated partial resistance to homologous challenge in calves and goats. Similar sensitization with F. gigantica or Schistosoma bovis also stimulated significant resistance to heterologous challenge in calves. The level of resistance was indicated by reduced fluke recoveries from sensitized animals compared with controls, by lower values of the enzymes sorbitol dehydrogenase and glutamate dehydrogenase, indicating less hepatic damage, and by lower blood indices, indicating more severe anaemia. Infection of calves with metacercariae of F. gigantica irradiated at 30 Gy or 200 Gy resulted in mean fluke recoveries of 75 and 41 respectively, whereas 332 flukes were recovered from controls infected with non-irradiated metacercariae. The flukes recovered from animals infected with irradiated metacercariae were stunted and resided in the small bile ductules. Sensitization for eight weeks with metacercariae of F. gigantica irradiated at 30 Gy resulted in significant resistance in calves, sheep and goats to homologous challenge. Significant resistance was also stimulated in calves by sensitization for eight weeks with metacercariae of F. gigantica irradiated at 200 Gy. However, sensitization for four weeks with F. gigantica irradiated at 30 Gy failed to stimulate significant resistance in calves to homologous challenge, but similar sensitization, twice at an interval of four weeks, resulted in an average reduction of 62% in fluke recovery from calves with homologous challenge. This reduction was, however, statistically insignificant. Moreover, sensitization for eight weeks with cercariae of Schistosoma bovis irradiated at 30 Gy did not stimulate heterologous resistance in calves or goats to F. gigantica. A similar result was also obtained after sensitization of calves for 36 weeks with cercariae of S. bovis irradiated at 30 Gy before challenge with F
Sohn, Woon-Mok; Na, Byoung-Kuk; Cho, Shin-Hyeong; Lee, Won-Ja; Park, Mi-Yeoun; Lee, Soon-Won; Choi, Seung-Bong; Huh, Beom-Nyung; Seok, Won-Seok
To know the infection status of zoonotic trematode metacercariae in brackish water fish, we surveyed mullets collected from 18 coastal areas in the Republic of Korea. The metacercariae of Pygidiopsis summa were detected in 236 (68.2%) out of 346 mullets examined. They were found in mullets from 15 areas except for those from Boseong-gun (Jeollanam-do), Pohang-si, and Uljin-gun (Gyeongsangbuk-do). Especially in mullets from Taean-gun (Chungcheongnam-do) and Geoje-si (Gyeongsangnam-do), their prevalences were 100% and 95.5%, and the average metacercarial density was more than 1,000 per fish. They were also detected in mullets from 3 coastal lakes, Gyeongpoho, Songjiho, and Hwajinpoho, in Gangwon-do, and their average densities were 419, 147, and 672 per infected fish, respectively. The metacercariae of 5 other heterophyid species, including Heterophyes nocens, Heterophyopsis continua, Metagonimus sp., Stictodora fuscata, and Stictodora lari, were found in the mullets examined. The metacercariae of H. nocens were detected in 66.7, 100, 28.6, 81.6, 3.9, 61.5, and 27.3% of mullets from Muan-gun, Shinan-gun, Haenam-gun, Gangjin-gun, and Boseong-gun (Jeollanam-do), Hadong-gun, and Geoje-si (Gyeongsangnam-do), and their metacercarial intensities were 64, 84, 119, 99, 1, 24, and 24 per fish infected, respectively. From the above results, it has been confirmed that P. summa metacercariae are heavily infected in mullets from coastal areas of Korea. It is suggested that residents who frequently consume raw mullet dish can be easily infected with heterophyid flukes.
Deng, Chuanhuan; Sun, Jiufeng; Li, Xuerong; Wang, Lexun; Hu, Xuchu; Wang, Xiaoyun; Chen, Wenjun; Lv, Xiaoli; Liang, Chi; Li, Wenfang; Huang, Yan; Li, Ran; Wu, Zhongdao; Yu, Xinbing; Xu, Jin
Aminopeptidases serve vital roles in metabolism of hormones, neurotransmission, turnover of proteins and immunological regulations. Leucine aminopeptidases catalyze the hydrolysis of amino-acid residues from the N-terminus of proteins and peptides. In the present study, leucine aminopeptidase 2 (LAP2) gene of Clonorchis sinensis (C. sinensis) was isolated and identified from an adult cDNA library of C. sinensis. Recombinant CsLAP2 was expressed and purified in Escherichia coli BL21. The open reading frame of LAP2 contains 1,560 bp equivalent to 519 amino acids, a similarity analysis showed a relatively low homology with Homo sapiens (19.0 %), Trypanosoma cruzi (18.0 %), Mus musculus (19.3 %), and relatively high homology with Schistosoma mansoni (65.6 %). The optimum condition of rCsLAP2 enzyme activity was investigated using a fluorescent substrate of Leu-MCA at 37 °C and pH 7.5. The K (m) and V (max) values of rCsLAP2 were 18.2 μM and 10.7 μM/min, respectively. CsLAP2 gene expression can be detected at the stages of the adult worm, metacercaria, excysted metacercaria and egg of C. sinensis using real-time PCR, no difference was observed at the stages of the adult worm, metacercaria and egg. However, CsLAP2 showed a higher expression level at the stage of excysted metacercaria than the adult worm (3.90-fold), metacercaria (4.60-fold) and egg (4.59-fold). Histochemistry analysis showed that CsLAP2 was located at the tegument and excretory vesicle of metacercaria, and the tegument and intestine of adult worm. The immune response specific to rCsLAP2 was characterized by a mixed response patterns of Th1 and Th2, indicating a compounded humoral and cellular immune response. The combined results from the present study indicate that CsLAP2 was an important antigen exposed to host immune system, and probably implicated as potential role in interaction with host cells in clonorchiasis.
Martínez-Aquino, Andrés; Aguilar-Aguilar, Rogelio; Pérez-Rodríguez, Rodolfo; de León, Gerardo Pérez-Ponce
Six helminth species were recorded during the helminthological examination of 35 specimens of the goodeid Xenotaenia resolanae from Arroyo Durazno, Jalisco, Mexico, a tributary of the Cuzalapa River. Helminth species identified included: 4 species of digeneans, i.e., Posthodiplostomum minimum (metacercariae), Clinostomum companatum (metacercariae), Dendrorchis sp. (adult), and Margotrema guillerminae (adult); and 2 species of nematodes, i.e., Spiroxys sp. (larvae) and Rhabdochona ahuehuellensis (adult). A very low number of individual larvae were found. The observed species richness, individual parasite abundance, and diversity were low at both component community and infracommunity levels. The values of similarity between infracommunities were relatively high because of the predominance of the digenean M. guillerminae, the species that reached the higher values of both prevalence and abundance. High water flow of the collecting site is suggested as the main factor determining the depauperate helminth assemblage in this fish species.
Qian, Bao-zhen; Sugiyama, H; Waikagul, J; Zhu, Zhi-hang
To identify Paragonimus harinasutai from Ninghai, Zhejiang Province, China. Metacercariae were collected from the crabs Sinopotamon chekiangenes in Xixi village of Ninghai County for ITS2 sequence analysis, CO1 sequence analysis and endonuclease BsaHI and StuI analysis by PCR-RFLP. Results The fingerprintings of PCR-RFLP were virtually same to the isolate from Thailand (Nakorn-nayok). The ITS2 sequence with 366 bp and CO1 sequence with 390 bp of the metacercariae collected from Ninghai revealed a nucleotide identity 95.6% and 89.5% respectively to the Thai isolate. The study confirmed that Paragonimus harinasutai is present in Ninghai, China, with certain variation on molecular biology in comparison to the Thai isolate.
Xu, Zheng-Min; Li, Ling; Wu, Xiao-Ying; Zhao, Han-Fen; Du, Ai-Ping; Hu, Sheng-Mei; Tao, Yong-Ping; Sun, Li; Tang, Yu-Cheng; Li, Ming-Hua; Zhang, Zhi-Yong; Li, Zhi-Shan
Freshwater crabs (Sinopotamon denticulatum) were examined for metacercariae. Cats and dogs were also examined for Paragonimus infection. Questionnairing was carried out on health knowledge and behaviors among local residents in a village of Baokang County, Hubei Province. Results showed that the infection rate of Paragonimus skrjabini metacercariae in Sinopotamon denticulatum was 20.5% (46/214), with 15.6% (20/128) in a mining area and 30.2% (26/86) for the non-mining area respectively (chi2 = 6.5, P 0.05). Questionnairing showed that dogs and cats were with the habit of foraging and defecating at streams and children had the habits of eating raw or under-cooked crabs. The natural and ecological environments are in favor of the life cycle of P. skrjabini.
Fredensborg, Brian Lund; Poulin, R
In this study, the effect of the composition of culture medium on excystation, growth, survival and egg production was investigated for the recently discovered microphallid trematode Maritrema novaezealandensis. Metacercariae from the two second intermediate crab hosts, Macrophthalmus hirtipes...... and Halicarcinus whitei, were incubated in either: (1) 0.85% saline solution, (2) the commercial cell culture medium, NCTC-109, (3) NCTC-109 supplemented with 20%, or (4) NCTC-109 supplemented with 40% chicken serum. Furthermore, excysted metacercariae were cultured for 5 days in each of the three media: NCTC-109...... cultured in media with a supplement of chicken serum and reached a maximum after 2 days of cultivation. Growth, however, did not occur after the first day of cultivation in any of the three media....
Kondo Yusuke; Ohtsuka Susumu; Hirabayashi Takeshi; Okada Shoma; Ogawa Nanako O.; Ohkouchi Naohiko; Shimazu Takeshi; Nishikawa Jun
In the Seto Inland Sea of western Japan, metacercariae of three species of trematodes, Lepotrema clavatum Ozaki, 1932, Cephalolepidapedon saba Yamaguti, 1970, and Opechona olssoni (Yamaguti, 1934), were found in the mesoglea of the jellyfish Aurelia aurita s.l., Chrysaora pacifica, and Cyanea nozakii. Moreover, these jellyfish frequently harbored juveniles of the fish species Psenopsis anomala, Thamnaconus modestus, and Trachurus japonicus. The former two fish species are well-known medusivor...
Nicanor Ibáñez H.
Full Text Available Se ha estudiado el proceso del desarrollo de Parogonimus peruvianus Miyazaki, Ibóñez y Miranda, 1969 en el gato doméstico, Felis cati L. infectado experimentalmente mediante la administración oral de metacercarias del parasito extraídas de ejemplares de Pseudothelphusa chilensis Milne Edwards, 1843 procedentes de áreas endémicas.
Mehrdana, Foojan; Jensen, Hannah M; Kania, Per W; Buchmann, Kurt
Ornamental fish, Xiphophorus maculatus, were imported from Singapore to Denmark for distribution to local aquarists. Importers observed lethargic and erratic swimming patterns among fish and forwarded a total of 30 fish for pathological examination to a university diagnostic service. All fish were diagnosed infected with encysted Centrocestus sp. metacercariae in gills (prevalence of 100% and mean intensity of 454.5 ± 161.9 parasites per fish). Metacercariae were identified by morphological and molecular methods. Cysts (mean length 163.3 ± 13.7 μm and mean width 113.3 ± 10.6 μm) contained a bent metacercaria with an X-shaped excretory bladder. PCR amplification of a rDNA region (5.8S rRNA gene, ITS-2, 28S rRNA gene) and subsequent sequencing confirmed the diagnosis. Metacercariae were found in gill filaments adjacent to the cartilage associated with cartilage hypertrophy, epithelial and mucous cell hyperplasia, clubbing and lamellar fusion. Host cell encapsulation of cysts comprised several layers of leucocytes, chondroblast-like and fibroblast-like cells. The observations raise concerns with regard to veterinary inspection and quarantine procedures. The zoonotic potential of these trematodes and a possible spread of the parasites in natural habitats in Europe should be regarded as a public health issue. So far, several cases of human infections have been reported only in Asia, but the potential intermediate host snail, Melanoides tuberculata, has been recorded in Germany. Accordingly, establishment of the parasite in Europe with climate changes should be considered a risk.
Faltýnková, Anna; Našincová, Vanda; Kablásková, Lenka
Roč. 14, č. 1 (2007), s. 39-51 ISSN 1252-607X R&D Projects: GA AV ČR IAA6022404; GA ČR GP524/07/P086; GA ČR GD524/03/H133; GA MŠk LC522 Institutional research plan: CEZ:AV0Z60220518 Keywords : Trematoda * Mollusca * cercaria * metacercaria * Lymnaea stagnalis Subject RIV: EG - Zoology Impact factor: 0.713, year: 2007
Nakao, Minoru; Waki, Tsukasa; Sasaki, Mizuki; Anders, Jason L; Koga, Daisuke; Asakawa, Mitsuhiko
In the Japanese Archipelago, Ezohelix gainesi, a member of bradybaenid land snails, is endemic mainly to the island of Hokkaido. During July to August of 2016, a survey to detect trematode infections from E. gainesi was carried out at a forest city park in Asahikawa, Hokkaido. Systemic infections of the snails with sporocysts containing short-tailed cercariae were found in 5.3% of 94 individuals examined. Furthermore, most of them (90.4%) harbored non-encysted metacercariae within their kidneys. A DNA sequence identification revealed that both of the sporocyst and the metacercaria belong to an unknown species of the family Brachylaimidae. The metacercariae showed a genetic diversity with 6 haplotypes of mitochondrial DNA (mtDNA) even in the limited sampling area. A definitive host of the unknown species could not be determined, although 34 field mice (Apodemus speciosus) and 21 voles (Myodes rufocanus) from the city park were examined for intestinal parasites. To examine the adult stage, the metacercariae were perorally administrated to mice, together with anti-inflammatory treatment with methylprednisolone. Fully matured adult worms were recovered from the intestinal ileum 8 and 14days postinfection. The gravid adults showed typical features of the genus Brachylaima. A morphological and biogeographical evaluation prompted us to propose Brachylaima ezohelicis sp. nov. for the parasite from E. gainesi. The autochthony of the first intermediate host and the spatial heterogeneity of mtDNA suggest that the new species found in the city park is not a recently expanded population of immigrant origin. Copyright © 2017 Elsevier B.V. All rights reserved.
Ma. del Carmen Guzmán-Cornejo; Luis García-Prieto
In September, 1995 and January, 1996, six species of fishes from Lake Cuitzeo, Michoacan, Mexico, were collected (Alloophorus robustus n=30, Goodea atripinnis n=30, Xenotoca variata n=41, Chirostoma jordani n=30, Carassius auratus n=30 and Oreochromis aureus n=30), to establish the infection levels reached by Posthodiplostomum minimum, Clinostomum complanatum and Diplostomum (Tylodelphys) sp., in their hosts. Infection caused by P. minimum metacercariae was the most important helminthiasis co...
TAJBAKHSH, Fatemeh; GOLZARIANPOUR, Kiavash; ABDOLI, Asghar
The present study was conducted to identify the parasitic infection of the nemacheilid loach, Turcinoemacheilus kosswigi, in 5 rivers of the Tigris basin in Iran, from July 2007 to August 2008. Extracted from 43 fish specimens were 3 parasitic species: metacercaria of Posthodiplostomum cuticola (Digenea: Diplostomatidae) in the skin, fins, gills, and muscles; Allocreadium sp. (Digenea: Allocreadidae) and Procamallanus sp. (Nematoda: Camallanidae) in the intestine. Prevalence and mean intensit...
Kudlai, Olena; Oros, M.; Kostadinova, Aneta; Georgieva, Simona
Roč. 10, 2 December (2017), č. článku 592. ISSN 1756-3305 R&D Projects: GA ČR GA15-14198S; GA ČR(CZ) GBP505/12/G112 Institutional support: RVO:60077344 Keywords : Diplostomum * Diplostomidae * Metacercariae * Freshwater fish es * Barcodes * cox1 * nad3 * River Danube * Europe Subject RIV: EG - Zoology OBOR OECD: Zoology Impact factor: 3.080, year: 2016
Sanpool, Oranuch; Intapan, Pewpan M; Thanchomnang, Tongjit; Janwan, Penchom; Nawa, Yukifumi; Blair, David; Maleewong, Wanchai
Paragonimiasis is an important food-borne parasitic zoonosis caused by infection with lung flukes of the genus Paragonimus. Of the 7 members of the genus known in Thailand until recently, only P. heterotremus has been confirmed as causing human disease. An 8th species, P. pseudoheterotremus, has recently been proposed from Thailand, and has been found in humans. Molecular data place this species as a sister species to P. heterotremus, and it is likely that P. pseudoheterotremus is not specifically distinct from P. heterotremus. In this study, we collected metacercariae of both nominal species (identification based on metacercarial morphology) from freshwater crabs from Phetchabun Province in northern Thailand, Saraburi Province in central Thailand, and Surat Thani Province in southern Thailand. In addition, we purchased freshwater crabs imported from Myanmar at Myawaddy Province, western Thailand, close to the Myanmar-Thailand border. The DNAs extracted from excysted metacercariae were PCR-amplified and sequenced for ITS2 and cox1 genes. The ITS2 sequences were nearly identical among all samples (99-100%). Phylogenies inferred from all available partial cox1 sequences contained several clusters. Sequences from Indian P. heterotremus formed a sister group to sequences from P. pseudoheterotremus-type metacercariae. Sequences of P. heterotremus from Thailand, Vietnam, and China formed a separate distinct clade. One metacercaria from Phitsanulok Province was distinct from all others. There is clearly considerable genetic variation in the P. heterotremus complex in Thailand and the form referred to as P. pseudoheterotremus is widely distributed in Thailand and the Thai-Myanmar border region.
Pérez-del-Olmo, A.; Georgieva, Simona; Pula, H.J.; Kostadinova, Aneta
Roč. 7, NOV 12 2014 (2014), s. 502 ISSN 1756-3305 R&D Projects: GA ČR GBP505/12/G112 Institutional support: RVO:60077344 Keywords : Diplostomum spathaceum * Diplostomum pseudospathaceum * Lens metacercariae * Freshwater fish * Gulls * Spain * Cox1 * TS1-5.8S-ITS2 Subject RIV: EG - Zoology Impact factor: 3.430, year: 2014
Chen, M.F.; Stewart, B.A.; Senkvik, Kevin; Hershberger, Paul
Nanophyetus salmincola is a parasitic trematode, or flatworm, that infects salmonid fishes in the Pacific Northwest, including Washington, Oregon, and portions of California. The adult worm lives in the intestine of fish-eating birds and mammals. Eggs shed into the water hatch into miracidia which penetrate the first intermediate host, one of two species of snail Juga plicifera or J. silicula. Asexual reproduction occurs within the snail. Free-swimming cercaria are released from the snail and penetrate the secondary intermediate host, often a salmonid fish, in fresh and brackish water. The cercaria encyst as metacercaria in various organs of the fish, including gills, muscle and heart, but favor the posterior kidney. Penetration and migration by the cercaria through the fish causes damage to nearly every organ system. Once encysted, metacercaria survive the ocean phase of salmonid life cycle. N. salmincola is a likely contributor to mortality of juvenile coho salmon (Oncorhynchus kisutch) during the early ocean rearing phase, and it is the most prevalent pathogen of outmigrating steelhead in the estuaries of the Pacific Northwest.A field survey was implemented from March-June 2014 to compare the prevalence and parasite load of N. salmincola infections in outmigrating steelhead from five Puget Sound watersheds and to assess changes in infection levels that occurred during the smolt out-migration through each watershed. N. salmincola infection prevalence and parasite loads were determined by counting metacercaria in posterior kidney samples. Tissue samples were collected and examined by standard histological methods.
Zhou, Ben-Jiang; Yang, Bin-Bin; Doanh, Pham Ngoc; Yang, Zhao-Qing; Xiang, Zheng; Li, Cui-Ying; Shinohara, Akio; Horii, Yoichiro; Nawa, Yukifumi
Among about 50 Paragonimus species, Paragonimus proliferus is a rare species characterized by extremely large metacercariae, most of which are present excysted in the crab hosts. Recently, this species was discovered by us in northern Vietnam as the first record outside of China. DNA sequences of both second internal transcribed spacer region (ITS2) and cytochrome oxidase subunit 1 gene (CO1) genes of the metacercariae and adult worms of P. proliferus of the Vietnamese isolates were identical with those of Paragonimus hokuoensis in the DNA database of the GenBank. To confirm those observations and to clarify the molecular phylogenetic status of P. proliferus, we determined the ITS2 and CO1 sequences of the metacercariae of P. proliferus obtained in Yunnan province, China where the original specimen was discovered. The results show that both ITS2 and CO1 sequences of P. proliferus of the Chinese isolates are identical with those of P. proliferus of the Vietnamese isolates and are also identical with those of P. hokuoensis that appeared in the DNA database (obtained in Yunnan province), suggesting the synonymy of P. hokuoensis with P. proliferus. By phylogenetic tree analyses, all samples of P. proliferus from China and Vietnam together with P. hokuoensis constructed a distinct group within, or very close to, Paragonimus skrjabini complex in both trees.
Sanpool, Oranuch; Intapan, Pewpan Maleewong; Thanchomnang, Tongjit; Janwan, Penchom; Laymanivong, Sakhone; Sugiyama, Hiromu; Maleewong, Wanchai
Paragonimus macrorchis is rather a rare species with sporadic discovery reports. To date, little is known about morphological features and the molecular phylogenetic status of P. macrorchis. Here we provide such information on P. macrorchis, of which metacercariae were collected from freshwater crabs in Khammouane Province, central Lao PDR. After morphological observation, metacercariae were excysted and were injected intra-peritoneally into Mongolian gerbils. Paragonimus adult worms were collected from the lungs of experimental gerbils 45 days after infection. A small piece of body tissue was cut at the posterior part of each adult worm for genomic DNA extraction. Then, the adult worms were stained and mounted for morphological identification. The second internal transcribed spacer region (ITS2) of rDNA and partial cytochrome c oxidase subunit 1 (cox1) gene were amplified using PCR method and sequenced. The results of morphological identification of metacercariae and adult worms together with their DNA sequences of ITS2 and partial cox1 gene clearly show that the specimens we collected in the central Lao PDR were P. macrorchis. Phylogenetic analyses revealed that P. macrorchis forms an independent cluster from other Paragonimus species in Asia. Copyright © 2015 Elsevier Ireland Ltd. All rights reserved.
Doanh, N Pham; Tu, A Luu; Bui, T Dung; Loan, T Ho; Nonaka, Nariaki; Horii, Yoichiro; Blair, David; Nawa, Yukifumi
Paragonimus westermani is one of the most medically important lung flukes and is widely distributed in Asia. It exhibits considerable variation in morphological, genetic and biological features. In central provinces of Vietnam, a high prevalence of metacercariae of this species has been reported from the crab intermediate host, Vietopotamon aluoiense. In this study, we detected P. westermani metacercariae in two additional crab hosts, Donopotamon haii in Quang Tri Province, central Vietnam and Indochinamon tannanti in Yen Bai Province in the north. The latter is a new locality for P. westermani in a northern region of Vietnam where P. heterotremus is the only species currently known to cause human paragonimiasis. Paragonimus westermani metacercariae found in Vietnam showed considerable morphological variation but slight genetic variation based on DNA sequences from the nuclear ribosomal ITS2 region and the mitochondrial 16S gene. Co-infection of the same individual crabs with P. westermani and P. heterotremus and/or some other Paragonimus species was found frequently, suggesting potential for co-infection in humans. The findings of the present study emphasize the need for highly specific molecular and immunodiagnostic methods to differentially diagnose between P. westermani and P. heterotremus infections.
Rangsiruji, Achariya; Sugiyama, Hiromu; Morishima, Yasuyuki; Kameoka, Yosuke; Donsakul, Thawat; Binchai, Sutheewan; Ketudat, Punsin
Field surveys of Paragonimus in Surat Thani Province, southern Thailand, revealed a new record of a lung fluke species other than P. westermani. The metacercariae were obtained from the crab, Ranguna smalleyi. The cysts of the metacercariae were spherical in shape and the larval body in the cysts contained pinkish granules. Fully mature adult worms were obtained from experimental infections with a rat and a ferret. The adult worms from the two host animals resembled each other, except for size, and had the anatomical characteristics of P. bangkokensis, ie the cuticular spines were arranged mainly in groups, the ovaries were highly branched, while the testes were more simply divided. Chromosomal preparations of the testes showed a haploid number of 11. As no sequence data of P. bangkokensis has been deposited in the GenBank/EMBL/DDBJ nucleotide database, the ITS2 region was sequenced using the metacercariae as starting material. A similarity search of P. bangkokensis ITS2 sequence using the BLAST program revealed that there was only one base difference between this population and P. harinasutai occurring in central Thailand. The result may suggest a close relationship between P. bangkokensis and P. harinasutai. This is the first description of Paragonimus species other than P. westermani occurring in southern Thailand.
Pinto, Hudson A.; Mati, Vitor L. T.; Melo, Alan L.
Fingerlings of Oreochromis niloticus collected in an artificial urban lake from Belo Horizonte, Minas Gerais, Brazil, were evaluated for natural infection with trematodes. Morphological taxonomic identification of four fluke species was performed in O. niloticus examined, and the total prevalence of metacercariae was 60.7% (37/61). Centrocestus formosanus, a heterophyid found in the gills, was the species with the highest prevalence and mean intensity of infection (31.1% and 3.42 (1–42), resp.), followed by the diplostomid Austrodiplostomum compactum (29.5% and 1.27 (1-2)) recovered from the eyes. Metacercariae of Drepanocephalus sp. and Ribeiroia sp., both found in the oral cavity of the fish, were verified at low prevalences (8.2% and 1.6%, resp.) and intensities of infection (only one metacercaria of each of these species per fish). These species of trematodes are reported for the first time in O. niloticus from South America. The potential of occurrence of these parasites in tilapia farming and the control strategies are briefly discussed. PMID:25485302
Jang, Du Hwan
The life cycle of Eurytrema pancreaticum has been studied locally at Chejudo (Quelpart Island) in Korea, and found a land snail, Acusta despecta Gray served as the first intermediate host of the pancreatic fluke. The second intermediate host of the fluke also has been surveyed there, the tettigoniid grasshoppers, Conocephalus maculatus Le Guillon and C. gladiatius Redtenbacher. The land snail, Acusta despecta Gray, is acquired infection with the eggs of the fluke in the autum, then the miracidia grow in the liver of the land snails and become mother sporocysts through the winter. After one mother sporocyst has been divided into many spindle shaped daughter sporocysts during spring session, the fully matured daughter sporocysts penetrate into the membrane of mantle cavity in the land snail, and then, pass actively the membrane between June and July. The daughter sporocysts are eaten by the tettigoniid grasshoppers, C. maculatus and C. gladiatus, through the summer and autumn. Cercariae in the grasshoppers grow for about 20 days, and become matured metacercariae in the abdominal cavity. Finally, the matured metacercariae together with the grasshoppers are eaten by their final hosts; goat, rabbit, etc. The artificial infection with the metacercariae of the pancreatic fluke to rabbit as the final host in laboratory room was carried out successfully. The miracidia have been forced out by pressure from the eggs incubated at 20 degrees ~ 25 degrees C could live more than three hours swimming around the medium under the microscope.
Khalil, Mokhtar Ibrahim; El-Shahawy, Ismail Saad; Abdelkader, Hussein Saad
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.
Muñoz, G; Valdivia, I; López, Z
We describe the life cycle of the bucephalid Prosorhynchoides carvajali from the intertidal rocky zone of central Chile. To elucidate the life cycle of this digenean, two mytilid bivalves, Semimytilus algosus and Perumytilus purpuratus, and ten intertidal fish species belonging to the families Blenniidae, Tripterygiidae, Labrisomidae, Kyphosidae and Gobiesocidae were analysed for natural infections. In addition, experimental infections of fish were undertaken and molecular analyses were performed of several developmental stages of the digeneans in various host species. Experimental infections of fish were made from infected mytilids to determine which fish species were suitable for the metacercarial stage of Prosorhynchoides. We also determined the abundance and prevalence of metacercariae in natural infections in fish and found that they were lower than in the experimental infections. A molecular analysis showed that sporocysts from S. algosus were identical to metacercariae from five fish species and P. carvajali adults. Sporocysts isolated from P. purpuratus were similar to metacercaria found in one fish species only (G. laevifrons) but were different from P. carvajali, with 1.9-2.0% genetic divergence. Therefore, the complete life cycle of P. carvajali consists of the mytilid species S. algosus as the first intermediate host, at least five intertidal fish species as second intermediate hosts (Scartichthys viridis, Auchenionchus microcirrhis, Hypsoblennius sordidus, Helcogrammoides chilensis and Gobiesox marmoratus), two carnivorous fish as definitive hosts (Auchenionchus microcirrhis and A. variolosus) and one occasional definitive host (Syciases sanguineus). This is the second description of a life cycle of a marine digenean from Chile.
Pinto, Hudson A; Mati, Vitor L T; de Melo, Alan L
The platynosomiasis, a worldwide parasitic disease with importance for domestic cat, has an etiological agent species of trematodes of the genus Platynosomum, whose complete life cycles are not yet known. The real role of lizards in the transmission of this dicrocoeliid parasite (as obligatory intermediate or paratenic host) still needs to be defined. In the present study, oval-shaped encysted metacercariae obtained from terrestrial isopods (Oniscidea sp. and Nagurus nanus) and elongated excysted metacercariae found in biliary ducts and gallbladder of lizards (Hemidactylus mabouia) in Brazil were used for morphological characterization and experimental infection of mice. Adult parasites recovered from bile ducts and liver of mice inoculated orally with metacercariae from both hosts (isopods and lizards) were identified as Platynosomum illiciens (=Platynosomum fastosum), showing that lizards are paratenic (not obligatory) hosts involved in the life cycle of this parasite. Moreover, Subulina octona is reported as the first intermediate host of P. illiciens in South America, and terrestrial isopods are presented here as new natural second intermediate hosts of the parasite. Finally, it is pointed out that high prevalence and intensity of infection of intermediate and paratenic hosts were observed. These findings on the life cycle of P. illiciens are relevant considering that they may indicate possible control measures of platynosomiasis.
Shimazu, Takeshi; Kino, Hideto
Metagonimus yokogawai (Katsurada, 1912) Katsurada, 1912 (Trematoda: Heterophyidae) is parasitic in the small intestine of mammals including man and birds in Far Eastern Russia, Korea, Japan, China, and Taiwan. In the present study, the metacercariae and adults of M. yokogawai were redescribed to designate a neotype of this fluke together with reviews of previous studies including study histories from the first discovery to now. We particularly, attempted to review the study histories and morphological descriptions of M. yokogawai for the species validity, and compared with the morphological characteristics and life cycles of the closely related species, Metagonimus takahashii and Metagonimus miyatai. Finally, we proposed a differential key for the 8 known Metagonimus species distributed in East Asia. Metacercariae were obtained from the body muscles of sweetfish (Plecoglossus altivelis) collected in the Asahi River at Takebe-cho, Kita-ku, Okayama City, Okayama Prefecture, Japan. Adults were recovered from the small intestine of Syrian golden hamsters, to which the metacercariae had been fed 14 days before. A neotype was selected out of the present adult specimens. The Asahi River at Takebo-cho became the type locality of M. yokogawai. In conclusion, the present review shows that M. yokogawai, M. takahashii, and M. miyatai are valid and discriminated by means of morphological characteristics. PMID:26537043
Pinto, H A; Melo, A L
Austrodiplostomum compactum has been involved in cases of ocular diplostomiasis in several species of fish in Brazil, but the molluscan intermediate hosts of the parasite remain unknown. In the present study, malacological surveys were carried out at Pampulha Reservoir, Belo Horizonte, Minas Gerais, Brazil, between January 2009 and July 2012. A total of 16,119 specimens of Biomphalaria spp. were collected and examined, of which 68/14,948 specimens (0.45%) of Biomphalaria straminea and 6/541 (1.11%) of Biomphalaria glabrata were found harboring a strigeid cercariae. Groups of 5 specimens of Cyprinius carpio were experimentally infected with these cercariae (100 larvae/fish), and metacercariae were recovered from the eyes of the fish, 65 days after infection, with a mean intensity of infection of 10.4 (8-13) metacercariae/fish. Morphological study on cercariae and metacercariae identified them as A. compactum . This is the first record of cercariae of A. compactum in Brazil, and B. straminea and B. glabrata as new intermediate hosts for the parasite.
Goncharov S. L.
Full Text Available The article describes occurrence and distribution of Cryptocotyle trematodes in fish in the waters of the Dnipro-Buh estuary and the Black Sea in Mykolaiv and Odesa Region. Study was conducted in 2015-2016. Two trematode species were found in natural waters of these regions: Cryptocotyle cancavum Crepli, 1825 and Cryptocotyle jejuna Nicoll, 1907. Th e latter species has not been previously registered in this region in southern Ukraine. Varying intensity of infection with Cryptocotyle metacercariae was observed in fish of Gobiidae family: Mesogobius batrachocephalus Pallas, 1814, Neogobius melanostomum Pallas, 1814, N. fluviatialis Pallas, 1814. Th e most affected species was N. melanostomum, with the prevalence of infection 59.2 %. Less infected were N. fluviatialis and M. batrachocephalus, with the prevalence of infection 30.4 % and 17 % respectively. The intensity of infection was the highest in N. melanostomum - 211 metacercariae per fish, followed by N. fluviatialis and M. batrachocephalu - 124 and 89 metacercariae respectively. Cryptocotyle was the most prevalent in the Dnipro-Buh estuary (cape Adzhigol, Mykolaiv Region and much less prevalent in the waters of the Black Sea in Mykolaiv and Odesa Regions. Mean prevalence of infection was 31.4 %.
Full Text Available Background: Human Echinostomiasis is an intestinal disease caused by the members of family Echinostomatidae parasites. The aim of present research was to identify echinostomatidae cercariae emitted by Lymnaea palustris snails from Mazandaran province in the north of Iran based on the morphological and morphometrical characteristics of the different stages of experimental parasite life cycle.Methods: Echinostomatidae cercariae were collected from L. palustris (Gastropoda: Lymnaeidae of the north of Iran. To collect metacercaria, 50 healthy snails were infected with cercariae experimentally (50 cercariae for each. To obtain the adult stage, 9 laboratory animals (3 ducks, 2 rats, 2 mice and 2 quails were fed with 60 metacercaria for each. To identify parasite, the different stages of worm were examined using light microscope and then the figures were draw under camera Lucida microscope and measures were determined.Results: Averagely, 15metacercaria were obtained from each snail that had been previously exposed with cercariae. Ducks presented worm eggs in feces after 10-15 days post-infection. Intestinal worms were collected and identified as Hypoderaeum conoideum on the bases of figures and measures of cephalic collar, the number of collar spine, suckers diameter ratio, testes arrangement, etc.Conclusion: H. conoideum cercariae and adult worm are described. This is the first report of the different stages of the experimental life cycle of this parasite in Iran.
Fasciola hepatica: comparative effects of host resistance and parasite intra-specific interactions on size and reproductive histology in flukes from rats infected with isolates differing in triclabendazole sensitivity.
Hanna, R E B; Gordon, A W; Moffett, D; Edgar, H W J; Oliver, L F; McConnell, S; Shaw, L; Brennan, G P; Fairweather, I
The efficacies of putative fasciolicides and vaccines against Fasciola hepatica are frequently monitored in clinical and field trials by determination of fluke egg output in host faeces and by worm counts in the host liver at autopsy. Less often used are parameters based on fluke size and histology, yet these can provide important indications of specific effects on the development of particular germ-line or somatic tissues, especially in relation to the timing and profligacy of egg production. In this study, F. hepatica metacercariae of two distinct isolates, the triclabendazole (TCBZ)-sensitive Cullompton isolate and the TCBZ-resistant Oberon isolate, were administered to rats as single-isolate or mixed-isolate infections. At autopsy 16 weeks later individual adult flukes were counted, measured and the reproductive organs were examined histologically. The degree of development of the testis tubules in each fluke was represented by a numerical score, based on the proportion of the histological section profiles occupied by testis tissue. The level of anti-F. hepatica antibody in the serum of each rat was determined by ELISA. It was found that Cullompton flukes were significantly larger than Oberon flukes, and that significantly more Cullompton metacercariae developed to adults than Oberon metacercariae. The Cullompton flukes showed histological evidence of aspermy and spermatogenic arrest, which was reflected in quantitatively reduced testicular development, as compared with the Oberon isolate. In Cullompton flukes, parthenogenetic egg development is implied. The size of Cullompton and Oberon flukes was significantly related to the number of adult flukes recovered, to the number of metacercariae administered, and to the percentage success of infection. The testis development score in both isolates was significantly related to the number of adult flukes recovered but not to the number of metacercariae administered, or to the percentage success of infection. Fluke
Cheng, You-Zhu; Li, Li-Sha; Lin, Guo-Hua; Zhou, Pei-Cong; Jiang, Dian-Wei; Fang, Yan-Yan; Lin, Chen-Xin; Li, Yan-Rong
To investigate the natural foci of paragonimiasis in Youxi, Yongtai and Pinghe Counties of Fujian Province. One village each from the three counties was selected according to reported paragonimiasis cases. Freshwater snails collected from fields were examined for cercariae. Freshwater crabs obtained in the fields were examined for metacercariae by washing filtration method or direct compression method. Feces of cats and dogs were collected for the detection of eggs by water precipitation. In order to identify the species of Paragonimus, 4 cats were orally fed with metacercariae. At the same time, the habitat of three survey sites was observed. Three fluke species, namely, P. skrjabini, P. westermani and P. cenocopiosus(Syn. Euparagonimus cenocopiosus) were found. In Banlin Village of Youxi County and Chishui Village of Yongtai County, the seropositive rate by IgG ELISA were 6.8% (21/309) and 6.8% (9/133), respectively. Four species of freshwater snails were found, two species of Tricula and one species of Pseudobythinella were newly identified first intermediate hosts of Paragonimus. Four species of freshwater carbs were found, one species of Nanhaipotamon served as a new second intermediate host of Paragonimus. In Youxi, the infection rate of P. skrjabini cercariae in snails and metacercariae in crabs was 2.1% (27/1 344) and 92.1% (58/63), respectively; the index of crab infection and the positive ratio of Paragonimus eggs in cat feces was 171.91 and 1/7, respectively. In Yongtai, the cercariae infection rate in Pseudobythinella and Tricula snails infected with P. skrjabini was 0.6% (4/690) and 0.1% (2/ 2330), respectively; the infection rate of P. skrjabini metacercariae in crabs was 18.2% (18/99); the index of crab infection was 9.12. In Pinghe, the infection rate of cercariae in Semisulcospira libertina and metacercariae in crabs was 03% (3/1092) and 44.9% (35/78), respectively; the index of crab infection was 616. Paragonimus skrjabini P. westermani and P
Lenis, Carolina; Galiano, Alicia; Vélez, Imelda; Vélez, Iván Darío; Muskus, Carlos; Marcilla, Antonio
Paragonimiasis is a subacute to chronic inflammatory granulomatous lung disease caused by the genus Paragonimus. In Latin America Paragonimus mexicanus Miyazaki & Ishii, 1968 is the only confirmed species to cause human infections. Paragonimus caliensis Little, 1968 is an uncommon species often regarded as a synonym of P. mexicanus. Recently, the study of two types of Paragonimus metacercariae from Costa Rica has provided new molecular and morphological evidence that P. caliensis is a separate species from P. mexicanus. In the present study, molecular, morphological and phylogenetic tools have been used to characterize two populations of Paragonimus located at west of Medellin, Antioquia and at Pichinde, Valle del Cauca (type locality of P. caliensis), Colombia. Adults and metacercariae obtained from Medellin, and metacercariae from Pichinde were analyzed. For morphological observations we used light microscopy and scanning electron microscopy (SEM). Morphology of metacercariae and adults matched with the holotype of P. caliensis. The number and arrangement of sensory papillae in the acetabulum region differs from the morphotypes reported for P. caliensis in Costa Rica. Two morphotypes in branching patterns of ovary and two morphotypes in branching patterns of testes were identified. The main morphological differences between P. caliensis and P. mexicanus corresponded to the size of gonads and their relative positions in the body, and the occasional presence of a cyst wall in P. caliensis metacercariae. The molecular and phylogenetic analyses (using nuclear ribosomal ITS2 and partial cytochrome c oxidase subunit 1 CO1 sequences) confirmed that P. caliensis from the type locality is the same species from Medellin and Costa Rica. Furthermore, these analyses also suggest genetic as well as geographical separation of P. caliensis populations between Colombia and Costa Rica. Currently, P. mexicanus and P. caliensis are sympatric in the Colombian Pacific bioregion, and
Ecological aspects of ectoparasites from the gills of Satanoperca pappaterra (Heckel, 1840 (Cichlidae from the upper Paraná river floodplain, Brazil = Aspectos ecológicos dos ectoparasitos branquiais de Satanoperca pappaterra (Heckel, 1840 (Cichlidae da planície de inundação do alto rio Paraná, Brasil
Gilberto Cezar Pavanelli
Full Text Available The gills of 34 specimens of Satanoperca pappaterra (Heckel, 1840 were analyzed, all of which had been collected from the upper Paraná river floodplain in Brazil, between March 2004 to June 2005. Five species of parasites were found: two digenean (metacercariae, only one monogenean, one cestode (plerocercoid and one copepod. The monogenean Sciadicleithrum sp. was the most abundant and prevalent, followed by the metacercariae Ascocotyle sp. The results did not show any significant difference in relation to host sex with regard to the prevalence and abundance of parasites between males and females. Only Sciadicleithrum sp. presented a significant positive correlation between standard host length and the abundance and prevalence ofparasitism. The metacercariae Ascocotyle sp. presented a significant positive relation only between standard host length and abundance. Through the results, a great abundance was observed forSciadicleithrum sp. and the metacercariae Ascocotyle sp. in connected lagoons. These differences suggest this type of environment can favor parasitism, especially in the case of monoxenic cycle ectoparasites, such as the monogenean. According to the importance value, only theSciadicleithrum sp. and the metacercariae Ascocotyle sp. were considered secondary, while the others were considered satellites. The typical pattern of overdispersion or aggregation from the parasitesystem was observed for Sciadicleithrum sp., the metacercariae Ascocotyle sp. and the metacercariae Diplostomum sp.Foram examinadas brânquias de 34 espécimes de Satanoperca pappaterra (Heckel, 1840, coletados na planície de inundação do Alto rio Paraná, Brasil, entre março de 2004 a junho de 2005. Foram encontradas cinco espécies de parasitos: dois digenéticos (metacercária, apenas um monogenético, um cestóide (plerocercóide e um copepoda. O monogenético Sciadicleithrum sp. foi a espécie mais abundante e a mais prevalente, seguida pela metacerc
Lee, Seon Kyu; Chang, Kee Hyun; Goo, Jin Mo; Han, Moon Hee; Shin, Yong Moon; Choo, Sung Wook; Yu, In Kyu; Cho, Seung Yull; Kong, Yoon
It is important to diagnosis paragonimiasis in early active because it can be dared by chemotherapy. However, it is difficult to make a correct diagnosis of cerebral paragonimiasis in the early active stage, and the radiographic findings of cerebral paragonimiasis have been rarely reported. Thus, this experimental study was designed to produce early active cerebral paragonimiasis and to demonstrate radiologic-pathologic correlations. In 8 cats, 7-8 metacercariae of Paragonimus Westermani were directly introduced into brain parenchyma of each cat's after trephination of the skull. In another 16 cats, the juvenile worms and the adult worms that had developed for varying periods (2 weeks, 4 weeks, 6 weeks, 8 weeks and 12 weeks) in the lunges of another cats were introduced into the brain parenchyma of each cat's with the same procedure described above. Follow -up MR images and chest radiographs were obtained at 2 days, 1 weeks, 2 weeks, 4 weeks and 8 weeks after inoculation. The autopsies and histopathological examinations of the cat's brain were undertaken in 22 cats. In 9 cats that were suspected with pulmonary lesion on chest radiograph, the soft tissue radiographs of inflated-fixed lungs were obtained. In one cat with inoculation of adult worm, acute suppurative inflammation of the brain parenchyma was demonstrated. But the other cats with inoculation of adult worm or juvenile worm and the cats with intentional of metacercaris did not reveal any evidence of acute cerebral paragonimiasis. More than half of the introduce metacercariae (5 out of 8 cats) were found in the lung parenchyma, while only 25% (4 out of 16 cats) of the adult worm inoculated cats were. Acute suppurative inflammation suggesting acute stage cerebral paragonimiasis was obtained in one case of adult worm inoculated cat. Most of the inoculated metacercariae and some of the juvenile worms or adult worms were migrated to the lungs
Lee, Seon Kyu; Chang, Kee Hyun; Goo, Jin Mo; Han, Moon Hee; Shin, Yong Moon; Choo, Sung Wook; Yu, In Kyu [Seoul National University College of Medicine, Seoul (Korea, Republic of); Cho, Seung Yull; Kong, Yoon [Chung-Ang University College of Medicine, Seoul (Korea, Republic of)
It is important to diagnosis paragonimiasis in early active because it can be dared by chemotherapy. However, it is difficult to make a correct diagnosis of cerebral paragonimiasis in the early active stage, and the radiographic findings of cerebral paragonimiasis have been rarely reported. Thus, this experimental study was designed to produce early active cerebral paragonimiasis and to demonstrate radiologic-pathologic correlations. In 8 cats, 7-8 metacercariae of Paragonimus Westermani were directly introduced into brain parenchyma of each cat's after trephination of the skull. In another 16 cats, the juvenile worms and the adult worms that had developed for varying periods (2 weeks, 4 weeks, 6 weeks, 8 weeks and 12 weeks) in the lunges of another cats were introduced into the brain parenchyma of each cat's with the same procedure described above. Follow -up MR images and chest radiographs were obtained at 2 days, 1 weeks, 2 weeks, 4 weeks and 8 weeks after inoculation. The autopsies and histopathological examinations of the cat's brain were undertaken in 22 cats. In 9 cats that were suspected with pulmonary lesion on chest radiograph, the soft tissue radiographs of inflated-fixed lungs were obtained. In one cat with inoculation of adult worm, acute suppurative inflammation of the brain parenchyma was demonstrated. But the other cats with inoculation of adult worm or juvenile worm and the cats with intentional of metacercaris did not reveal any evidence of acute cerebral paragonimiasis. More than half of the introduce metacercariae (5 out of 8 cats) were found in the lung parenchyma, while only 25% (4 out of 16 cats) of the adult worm inoculated cats were. Acute suppurative inflammation suggesting acute stage cerebral paragonimiasis was obtained in one case of adult worm inoculated cat. Most of the inoculated metacercariae and some of the juvenile worms or adult worms were migrated to the lungs.
Full Text Available This study of infection of southern Australian land snails with Brachylaima cribbi metacercariae has shown that all commonly encountered native and introduced snails are susceptible second intermediate hosts. The range of infected snails is extensive with metacercariae-infected snails being present in all districts across southern Australia. C. virgata has the highest average natural metacercarial infection intensity of 6.1 metacercariae per infected snail. The susceptibility of birds, mammals and reptiles to B. cribbi infection was studied in South Australia by capturing, dissecting and examining the intestinal tract contents of animals which commonly eat land snails as a food source. Indigenous Australian little ravens (Corvus mellori, which are a common scavenger bird, and two other passeriform birds, the black bird (Turdus merula and the starling (Sturnus vulgaris, which are both introduced European birds, were found to have the highest infection rates of all animals examined. Other birds found infected with B. cribbi were an emu (Dromaius novaehollandiae, chickens (Gallus gallus and a pigeon (Columba livia. Natural infections were also detected in field mice (Mus domesticus and shingleback lizards (Tiliqua rugosa although the intensity of infection was lower than that observed in birds. Susceptibility studies of laboratory mice, rats and ducks showed that mice developed patent infections which persisted for several weeks, rats developed a short-lived infection of three weeks’ duration and ducks did not support infection. This study has shown for the first time that a brachylaimid can infect a wide host range of birds, mammals and reptiles in nature.
Blasco-Costa, I; Poulin, R; Presswell, B
Among eyeflukes, Tylodelphys Diesing, 1850 includes diverse species able to infect the eyes, but also the brain, pericardial sac and body cavity of their second intermediate host. While the genus shows a cosmopolitan distribution with 29 nominal species in Africa, Asia, Europe and America, a likely lower research effort has produced two records only for all of Australasia. This study provides the first description of a species of Tylodelphys and the first record for a member of the Diplostomidae in New Zealand. Tylodephys sp. metacercaria from the eyes of Gobiomorphus cotidianus McDowall, 1975 is distinguished from its congeners as being larger in all, or nearly all, metrics than Tylodelphys clavata (von Nordmann, 1832), T. conifera (Mehlis, 1846) and T. scheuringi (Hughes, 1929); whereas T. podicipina Kozicka & Niewiadomska, 1960 is larger in body size, ventral sucker and holdfast sizes and T. ophthalmi (Pandey, 1970) has comparatively a very small pharynx and body spination. Tylodelphys sp. exhibits consistent genetic variation for the 28S rDNA, internal transcribed spacer (ITS) and Cox1 genes, and phylogenetic analyses confirm that it represents an independent lineage, closely related to North American species. Morphological and molecular results together support the distinct species status of Tylodephys sp. metacercaria, the formal description and naming of which await discovery of the adult. Furthermore, the validity of T. strigicola Odening, 1962 is restored, T. cerebralis Chakrabarti, 1968 is proposed as major synonym of T. ophthalmi, and species described solely on the basis of metacercariae are considered incertae sedis, except those for which molecular data already exist.
Hong, Sung Jong; Lee, Soon Hyung; Seo, Byong Seol; Hong, Sung Tae; Chai, Jong Yil
An experimental study was carried out to observe the susceptibility of several kinds of laboratory animals to Fibricola seoulensis infection, a diplostomatid fluke of mammals. The metacercariae were obtained from the viscera of the snakes, Natrix tigrina lateralis and 50-2,000 in number each was artificially fed to a total of 127 animals; albino rats, mice, dogs, cats, rabbits and chickens. After 3 days to 8 weeks the animals were sacrificed and the recovery rate of worms as well as their maturity was observed. The results are as follows: The overall wom recovery rates throughout the experimental period was highest in albino rats(40.0 %) followed by mice(33.9%), cats(20.9 %), dogs(11.4 %), rabbits(0.05 %) and chickens(0 %). However, the recovery rates in the same host decreased as infection progressed longer and variable by the amount of metacercariae given. From albino rats and mice, the highest recovery rates were obtained in 1,000 and 200 metacercariae infection groups repectively, and it is considerd that such amount should be the optimum dose for experimental infection of these animals. The main location of F. seoulensis in experimental animals was small intestine especially the duodenum. The maturity index (No. mature worms/No. examined) was 100% in albino rats and mice, while only 22.7% or 0% in dogs or cats respectively. From the results, it is concluded that albino rats and mice are the most susceptible hosts for F. seoulensis infection among six kinds of laboratory animals examined.
Dreyfuss, G; Vignoles, P; Rondelaud, D
Experimental co-infections of juvenile and pre-adult Pseudosuccinea columella with Calicophoron daubneyi and Fasciola hepatica (five miracidia of each digenean per snail) were carried out to determine the aptitude of this lymnaeid to ensure complete larval development of the former parasite, the latter or both. Snails infected with F. hepatica were found in the two groups of juveniles, i.e. 1 and 2 mm at exposure, and the four groups of pre-adults, i.e. 3-6 mm. The highest frequency of F. hepatica, i.e. 37.3%, was noted in the 4 mm group. Low frequencies were noted for C. daubneyi and co-infections of both digeneans in the 3, 4 and 5 mm groups. Two other groups of P. columella, measuring 3 and 4 mm at exposure, were also constituted to study the characteristics of these co-infections. Compared to controls infected only with F. hepatica, the frequency of this digenean infection and the mean number of metacercariae were significantly lower in co-infected snails, while the patent period was significantly shorter. In snails harbouring C. daubneyi only or both digeneans, lower values were noted for prevalence, the patent period and the number of metacercariae. Pre-adult P. columella (3-5 mm in shell height at exposure) were able to sustain larval development of C. daubneyi if they were co-infected with the sequence C. daubneyi +F. hepatica. Low values noted for the prevalence of C. daubneyi infection and the number of metacercariae would be in favour of a still incomplete adaptation between the snail population and the miracidial isolate.
Odermatt, P; Habe, S; Manichanh, S; Tran, D S; Duong, V; Zhang, Wei; Phommathet, K; Nakamura, S; Barennes, H; Strobel, M; Dreyfuss, G
Human cases of paragonimiasis have been reported in Lao People's Democratic Republic from several provinces, however, detailed descriptions of a focus of transmission including parasites species and intermediate hosts are lacking. Field investigations were carried out between February and July 2004 in three neighboring villages of the province of Vientiane. Persons with chronic cough (more than three weeks) were identified during a door-to-door survey and three sputum samples were examined for Paragonimus spp. and tuberculosis on two consecutive days. Potential intermediate snail and crustacean hosts were sampled among the populations' water-contact sites and examined for metacercarial infections or cercarial shedding. Four dogs were infected with metacercaria and the species of the subsequently retrieved adult parasites were identified. 12.7% (15 cases) of all chronic cough patients (0.6% of the total population) suffered from paragonimiasis. Samples of five species of freshwater snails, two species of crabs and one species of prawn were collected from the Nam Sêt River. None of the snails were infected. Metacercariae of P. harinasutai, P. bangkokensis and P. heterotremus were found in Potamon lipkei crabs while an infection with P. westermani was observed in a Chulathelphusa brandti crab. No paragonimid metacercaria was found in the Macrobrachium dienbienphuense prawns Dang and Nguyen 1972. This study demonstrates the importance of Paragonimus spp. as an etiological agent in patients with chronic cough and suggests that similar transmission might be widespread in Lao People's Democratic Republic. We document herein the transmission of several Paragonimus species indicating that paragonimiasis is an active zoonosis in Lao People's Democratic Republic.
Villa-Mancera, A; Quiroz-Romero, H; Correa, D; Alonso, R A
The objective of this study was to assess the proteolytic activity of Fasciola hepatica cathepsins in liver sections from mice vaccinated with phage clones of cathepsin L mimotopes, using the film in situ zymography technique. Female BALB/c mice were immunized three times with 2.5 x 10¹¹ phage particles without adjuvant. Animals vaccinated with phage clones produced high titres of anti-mimotope antibodies and a significant reduction in fluke burden was observed following challenge with metacercariae of F. hepatica. The proteolytic activity in hepatic tissue was reduced after the immunization with phage clones.
Lu, Yu-Yun; Liu, Qiao; Tang, Gao-Xing; Shen, Hao-Xian; Zhong, Jian-Xin; Xie, Quan-Chao; Fu, Guang-Hua; Chen, Yu-Lian; Li, Hao-Bin; He, Hai-Feng
To investigate the current distribution of Paragonimus westermani in Guangdong Province. Snails and crabs collected from mountain streams in regional survey sites were dissected to detect cercarial and metacercarial infections of P. westermani. Domestic cats and dogs artificially infected with the collected metacercariae were also dissected to detect adult worms of P. westermani. The COI and ITS2 gene sequences of those adult worms were compared with those of known Paragonimus specimen deposited in the GenBank. All of the first intermediate hosts in five survey sites of Liangkou, Nankun, Mountain, Dadong, Muxi, Guowu, were identified as Semisulcospira libertina, whose cercariae infection rates were 0.33%, 0.15%, 0.058%, 0.10%, and 0.05%, respectively; the second intermediate hosts in above five sites were all identified as Sinopotamon denticulatum, whose metacercariae infection rates were 100%, 100%, 38.09%, 55.36%, and 65.26%, respectively. The numbers of metacercariae in the five sites were 79.4, 105.66, 9.16, 16.18, and 15.6 per positive crab, respectively, and 11.12, 7.87, 0.58, 0.69, and 0.85 per gram of crab, respectively. All the metacercariae were identical to those of P. westermani. Adult worms and eggs of P. westermani were found in both reservoir hosts of domestic cats and dogs infected artificially. By comparing the COI genes of five representative samples from each survey site with that of Paragonimus #AF219379.21, AF540958.1 from GenBank, we found out the homology to be 99%, 99%, 99%, 98%, and 99%, respectively. In addition, a comparison of the ITS2 gene sequences between the above five samples and Paragonimus #DQ836243.1, DQ351845.1, AB354217.1 from GenBank revealed 98%, 99%, 98%, 98%, and 98% gene homology, respectively. Two ultra-high and three high endemic areas of P. westermani are discovered in Guangdong Province. No obvious differences were found among the types of P. westermani in the above five endemic areas.
Lou, Hong-Qiang; Hu, Ye; Jin, Yao-Jian; Yu, Xin-Tu; Wang, Lan; He, Xu-Ying; Tu, Ping-Guang
To investigate the natural hosts infected with Paragonimus sp. and identify the species of the parasite in selected counties/districts of Jinhua prefecture in Zhejiang Province. Three townships/towns were randomly sampled from each of the 9 counties/districts in Jinhua as pilot spots for the survey. Fresh-water snails were collected from the fields for examining cercariae. Crabs were collected and detected for metacercariae by routine technique and the metacercariae were fed to dogs purchased in areas free from paragonimiasis. Fecal materials of dogs and cats around the villages and streams where crabs were found infected were collected for examining eggs. The artificially infected dogs were sacrificed 55 d after infection to receive adult worms. The size of cercariae, metacercariae, eggs and adult worms was measured. After the DNA of the adult worm was extracted, PCR was used to amplify the COI gene and ITS2 gene of the mitochondria from the worms. Homology with relative strains/isolates was analyzed and phylogenetic tree constructed. The survey demonstrated that the snail Semisulcospira libertina and the crab Sinopotamon chekiangense served as the first and second intermediate hosts respectively. Natural infection was found in Wucheng District with an infection rate of 0.2% (2/1,088) in snails and 76.7% (46/60) in crabs in Shafan township, and an infection index (II) of 2.0 in crabs, 0.1% (1/1,683) in snails and 53.0% (46/60) in crabs with an II of 0.9 in Langya town. The infection rate was 0 (0/575) in snails and 30.0% (18/60) in crabs with an II of 0.1 in Baimu township of Wuyi County. Paragonimus eggs were detected in feces of stray cats with a positive rate of 8.3% (1/12) in Shafan and 0.6% (1/17) in Langya. The size and morphology of the cercariae, metacercariae, eggs and adult worms were similar to those of Paragonimus westermani. The sequences of the COI and ITS2 genes were with 390 bp and 363 bp respectively, indicating a homology of 88.2%-98.2% and 86
Full Text Available Twelve Tuli weaner steers aged 1 year were randomly subdivided into three groups of four animals and infected with different doses of Calicophoron microbothrium metacercariae. Each animal in Group I received a low dose (LD of 5 000 metacercariae, Group II a medium dose (MD of 15 000 metacercariae, Group III a high dose (HD of 25 000 metacercariae and one additional animal was kept as an uninfected control (C. After infection, one animal from each group was slaughtered on Day 28, 42, 56 and 84 post infection (pi and samples from the ileum, jejunum, duodenum, abomasum and the rumen were collected for histopathological and cytological examination. On Day 28 pi, the gross pathological lesions observed in the duodenum of the LD and the MD animals were similar and comprised duodenal thickening, corrugation, hyperaemia, petechiation and ulceration. In the HD animal the duodenal lesions were similar but more severe. The abomasal folds were severely oedematous in the MD group and nearly occluded the abomasal lumen. Moderate oedema of the abomasal folds was also present in the LD and HD animals. The gross pathological lesions regressed in all the infected groups with increasing age of infection and had disappeared completely by Day 56 pi. On Day 28 pi the histopathological lesions in the duodenum and jejunum of the LD and MD groups were similar, comprising subtotal villous atrophy, hyperplasia of Brunner's glands and Peyer's patches and moderate infiltration of eosinophils, mast cells and a few globule leukocytes, basophils and lymphocytes in the lamina propria. The HD group had total villous atrophy, severe hyperplasia and cystic dilatation of Brunner's glands, which had expanded to cover the entire submucosa. On Day 42 pi the histopathological lesions were still present in the MD and the HD groups comprising subtotal villous atrophy and hyperplasia of Brunner's glands. Heavy infiltrations of eosinophils, moderate amounts of mast cells and a few
Margarita Ostrowski de Nunez
Full Text Available The life cycle of Ascocotyle (Leighia hadra n.sp. was experimentally reproduced, starting from cercariae from naturally infected Littoridina parchappei, collected from Los Ranchos stream, near Mercedes city, Buenos Aires Province, Argentina. Metacercariae were found encysted in the liver and mesentery of experimentally and naturally infected fishes Cnesterodon decemmaculatus and Jenynsia lineata. Adults were obtained experimentally in chicks and mice. The natural host is unknown. The new species is compared with Ascocotyle (Leighia mcintoshi Price 1936 as described by Leigh, 1974, differing in behavior and morphology of cercarial, metacercarial and adult stages
The effect of varying doses (1 kr, 3 kr, 5 kr) of x-irradiation on the development of Fasciola gigantica, the common liver fluke of livestock in India has been investigated in goat kids with a view to develop a radiation attenuated vaccine against the pathogen. The growth and development of the flukes recovered from animals infected with irradiated metacercarie is found to be retarded. Metacecariae exposed to 3 kr can develop further, but number of flukes recovered from animals infected with metacecariae exposed to 5 kr is considerably less than the one from animals infected with non-irradiated metacercariae. (M.G.B.)
Sirgel Wilhelm F
Full Text Available Abstract Background The life cycle of the brachylaimid trematode species Renylaima capensis, infecting the urinary system of the shrew Myosorex varius (Mammalia: Soricidae: Crocidosoricinae in the Hottentots Holland Nature Reserve, South Africa, has been elucidated by a study of its larval stages, epizootiological data in local snails and mammals during a 34-year period, and its verification with mtDNA sequencing. Methods Parasites obtained from dissected animals were mounted in microscope slides for the parasitological study and measured according to standardized methods. The mitochondrial DNA cox1 gene was sequenced by the dideoxy chain-termination method. Results The slugs Ariostralis nebulosa and Ariopelta capensis (Gastropoda: Arionidae act as specific first and second intermediate hosts, respectively. Branched sporocysts massively develop in A. nebulosa. Intrasporocystic mature cercariae show differentiated gonads, male terminal duct, ventral genital pore, and usually no tail, opposite to Brachylaimidae in which mature cercariae show a germinal primordium and small tail. Unencysted metacercariae, usually brevicaudate, infect the kidney of A. capensis and differ from mature cercariae by only a slightly greater size. The final microhabitats are the kidneys and ureters of the shrews, kidney pelvis and calyces in light infections and also kidney medulla and cortex in heavy infections. Sporocysts, cercariae, metacercariae and adults proved to belong to R. capensis by analysis of a 437-bp-long cox1 fragment, which was identical except for three mutations in metacercariae, of which only one silent. Epizootiological studies showed usual sporocyst infection in A. nebulosa and very rare metacercarial infection in A. capensis, which does not agree with high prevalences and intensities in the shrews. Conclusions The presence of monotesticular adult forms and larval prevalences and intensities observed suggest that R. capensis may use two transmission
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...
Clausen, Jesper Hedegaard; Madsen, Henry; Murrell, Kenneth Darwin
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......Fish-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...
Ruiz, M; Darriba, S; Rodríguez, R; López, C
This paper reports the results of the survey developed after the first detection of protozoan Marteilia sp. infection of the grooved razor shell Solen marginatus (Pulteney, 1799) from Galicia (NW Spain) in 2006. Furthermore, we analysed other parasites and pathological conditions found in grooved razor shell populations throughout this survey, such as metacercariae of trematodes, prokaryotic infections and disseminated neoplasms, some of which could cause moderate or severe damage to the host depending on the intensity of infection. A total of 17 natural beds distributed along the Galician coast were analysed, and Marteilia sp. was detected in 6 of them with low prevalence, moderate intensity and no negative effects over the populations.
Susana Balmant Emerique Simões
Full Text Available The life history of the trematode Pygidiopsis macrostomum Travassos, 1928 is described for the first time. Rediae and cercariae were obtained from naturally infected snails Heleobia australis (d´Orbigny, a new first intermediate host. Metacercariae were found encysted in the mesenteries of three naturally infected guppies, Phalloptychus januarius (Hensel, Jenynsia multidentata (Jenyns (new host records and Poecilia vivipara Bloch and Schneider. Experimental infections were successfully completed in the intermediate hosts H. australis and Poe. vivipara reared in the laboratory and hamsters Mesocricetus auratus Waterhouse were utilised as a definitive host.
Pérez, J; Martín de las Mulas, J; Carrasco, L; Gutierrez, P N; Martínez-Cruz, M S; Martínez-Moreno, A
Lesions produced by Fasciola hepatica in the liver, gall-bladder and hepatic lymph nodes (HLNs) of four groups of five goats are described; in addition, the distribution of CD3+ T lymphocytes and IgG-lambda light chain-bearing cells was analysed in the hepatic lesions and HLNs. One group of goats received a single oral dose of metacercariae, but the other four groups received four or five doses at different intervals over a period of 11 weeks. Animals that survived were killed 53-55 weeks after the first infective dose. Goats were more susceptible to multiple doses than to a single dose, even when the total number of metacercariae was the same. This greater susceptibility was manifested by the occurrence of deaths and the severity of hepatic lesions. CD3+ lymphocytes were sparse in the infiltrate surrounding the acute migratory tracts, suggesting inhibition of the local cell-mediated immune response. Goats with numerous hepatic calcareous granulomas showed the most severe hepatic damage, including marked cirrhosis, with a striking infiltrate of CD3+ T lymphocytes and lambda IgG- plasma cells, replacing extensive areas of hepatic parenchyma, in which hypertrophy of the smooth endoplasmic reticulum of hepatocytes was evident. These findings were observed mainly in the goats given more than one infective dose.
In-vivo monitoring of development of cholangiocarcinoma induced with C. sinensis and N-nitrosodimethylamine in Syrian golen hamsters using ultrasonography and magnetic resonance imaging: a preliminary study
Woo, Hyunsik [SMG-SNU Boramae Medical Center, Department of Radiology, Seoul (Korea, Republic of); Han, Joon Koo; Kim, Jung Hoon [Seoul National University Hospital, Department of Radiology, Seoul (Korea, Republic of); Hong, Sung-Tae [Seoul National University, Department of Parasitology, College of Medicine, Seoul (Korea, Republic of); Uddin, M.H. [Seoul National University, Adult Stem Cell Research Center, Laboratory of Stem Cell and Tumor Biology, Department of Veterinary Public Health, College of Veterinary Medicine, Seoul (Korea, Republic of); Jang, Ja-June [Seoul National University, Department of Pathology, College of Medicine, Seoul (Korea, Republic of)
The purpose of this study is to evaluate high-resolution ultrasound and magnetic resonance imaging (MRI) in monitoring of cholangiocarcinoma in the hamsters with C. sinensis infection and N-nitrosodimethylamine (NDMA). Twenty-four male Syrian golden hamsters of were divided into four groups composed of five hamsters as control, five hamsters receiving 30 metacercariae of C. sinensis per each hamster, five hamsters receiving NDMA in drinking water, and nine hamsters receiving both metacercariae and NDMA. Ultrasound was performed every other week from baseline to the 12th week of infection. MRI and histopathologic examination was done from the 4th week to 12th week. Cholangiocarcinomas appeared as early as the 6th week of infection. There were 12 cholangiocarcinomas, nine and ten of which were demonstrated by ultrasound and MRI, respectively. Ultrasound and MRI findings of cholangiocarcinomas in the hamsters were similar to those of the mass-forming intrahepatic cholangiocarcinomas in humans. Ultrasound and MRI also showed other findings of disease progression such as periductal increased echogenicity or signal intensity, ductal dilatation, complicated cysts, and sludges in the gallbladder. High-resolution ultrasound and MRI can monitor and detect the occurrence of cholangiocarcinoma in the hamsters non-invasively. (orig.)
Ngoc Doanh, Pham; Shinohara, Akio; Horii, Yoichiro; Yahiro, Shinichiro; Habe, Shigehisa; Vannavong, Nanthasane; Strobel, Michel; Nakamura, Satoshi; Nawa, Yukifumi
Paragonimus bangkokensis and Paragonimus harinasutai were found one after another in the same crab host, Potamon smithianus, in Thailand in 1967 and 1968. Both species were also recently found in China and Lao People's Democratic Republic (PDR). Those two Paragonimus spp. are distinguishable from each other by morphological features of metacercariae and adults. However, recently, the DNA sequences of second internal transcribed spacer region (ITS2) and cytochrome oxidase subunit 1 (CO1) genes of those two species in Thailand were reported to be highly similar to each other. In the present study, we collected P. bangkokensis in two provinces in Vietnam (the first record in Vietnam) and both P. bangkokensis and P. harinasutai in Lao PDR for the morphological and molecular phylogenetic analyses to clarify the mutual relationship between the two species. The results show that P. bangkokensis and P. harinasutai were distinguishable from each other by morphology such as the size of metacercariae and the arrangement of cuticular spines of adult worms. However, the molecular phylogenetic analyses of ITS2 and CO1 genes clearly indicate that P. bangkokensis and P. harinasutai make a monophyletic group.
Singh, T Shantikumar
To determine the natural crustacean host, the Paragonimus species, and to investigate the potential host-parasite relationship between Manipur Paragonimus and some of the laboratory animals. The laboratory animals such as puppies, albino rats and Swiss mice were infected orally with metacercariae isolated from the fresh water crabs, Potamiscus manipurensis. The fecal specimens of the experimentally infected animals were examined microscopically for Paragonimus eggs at regular intervals. The animals were autopsied on days 35 to approximately 328 after infection and the isolated worms were flattened between glass slides and fixed in 70% alcohol. The worms were stained with carmine and mounted with Canada balsam for morphological studies. The eggs were collected in 5% formol saline solution for microscopy. The flukes were classified into 4 developmental stages. A total of 11 worms, 5 mature, 5 immature and 1 pre-adult were recovered. The morphological features of the metacercariae, worms and eggs were similar to those of Paragonimus heterotremus. Manipur is one of the rare areas in the world where Paragonimus heterotremus is prevalent and the puppies are ideal experimental animal host. This species may be one of the important causes of paragonimiasis in animals and humans in Manipur.
Singh, T S; Sugiyama, H; Umehara, A; Hiese, S; Khalo, K
To determine the prevalence of paragonimiasis among the patients who were attending the tuberculosis (TB) clinics at the Community Health Centre, Pfutsero, Phek District, Nagaland. To determine the species of Paragonimus that cause infection in humans and the crustacean host that acts as the infectious source for humans. Sputum specimens were examined microscopically for Paragonimus eggs and acid fast bacilli. Blood samples were tested by microenzyme-linked immunosorbant assay for Paragonimus-specific immunoglobulin G antibodies. Crab extracts prepared by digestion with artificial gastric juice were examined for Paragonimus metacercariae under a stereoscopic microscope. The species identification of the parasite was based on morphological and molecular characterizations of eggs and metacercariae employing polymerase chain reaction and DNA sequencing. Seven out of the 14 patients tested seropositive for paragonimiasis and Paragonimus eggs were detected in sputum of two out of the seven seropositive patients, indicating a prevalence of 50% and an egg detection rate of 14%, respectively. The prevalence was highest in the 10-30 year age group. More males got the infection than females, the ratio being 5:2. P. heterotremus was identified as the causative agent of human paragonimiasis and Potamiscus manipurensis as the crab host. The study revealed that paragonimiasis has been endemic in Pfutsero, Nagaland, and half of the patients attending the TB clinic were actually suffering from pulmonary paragonimiasis. This is the first confirmed report of an endemic focus of paragonimasis and description of P. heterotremus as the causative agent in Nagaland, India.
In-vivo monitoring of development of cholangiocarcinoma induced with C. sinensis and N-nitrosodimethylamine in Syrian golen hamsters using ultrasonography and magnetic resonance imaging: a preliminary study
Woo, Hyunsik; Han, Joon Koo; Kim, Jung Hoon; Hong, Sung-Tae; Uddin, M.H.; Jang, Ja-June
The purpose of this study is to evaluate high-resolution ultrasound and magnetic resonance imaging (MRI) in monitoring of cholangiocarcinoma in the hamsters with C. sinensis infection and N-nitrosodimethylamine (NDMA). Twenty-four male Syrian golden hamsters of were divided into four groups composed of five hamsters as control, five hamsters receiving 30 metacercariae of C. sinensis per each hamster, five hamsters receiving NDMA in drinking water, and nine hamsters receiving both metacercariae and NDMA. Ultrasound was performed every other week from baseline to the 12th week of infection. MRI and histopathologic examination was done from the 4th week to 12th week. Cholangiocarcinomas appeared as early as the 6th week of infection. There were 12 cholangiocarcinomas, nine and ten of which were demonstrated by ultrasound and MRI, respectively. Ultrasound and MRI findings of cholangiocarcinomas in the hamsters were similar to those of the mass-forming intrahepatic cholangiocarcinomas in humans. Ultrasound and MRI also showed other findings of disease progression such as periductal increased echogenicity or signal intensity, ductal dilatation, complicated cysts, and sludges in the gallbladder. High-resolution ultrasound and MRI can monitor and detect the occurrence of cholangiocarcinoma in the hamsters non-invasively. (orig.)
Full Text Available As large numbers of metacercariae of Fasciola hepatica are necessary for research, experimental infections of Galba truncatula and Pseudosuccinea columella with this digenean were carried out to determine the better intermediate host for metacercarial production and, consequently, the most profitable snail for decreasing the cost price of these larvae. Pre-adult snails (4 mm in shell height originating from two populations per lymnaeid species were individually exposed to two or five miracidia, raised at 23 °C and followed for cercarial shedding up to their death. Compared to values noted in G. truncatula, the survival of P. columella on day 30 post-exposure was significantly greater, while the prevalence of F. hepatica infection was significantly lower. In the four P. columella groups, metacercarial production was significantly greater than that noted in the four groups of G. truncatula (347–453 per cercariae-shedding snail versus 163–275, respectively. Apart from one population of G. truncatula, the use of five miracidia per snail at exposure significantly increased the prevalence of F. hepatica in P. columella and the other population of G. truncatula, whereas it did not have any clear effect on the mean number of metacercariae. The use of P. columella for experimental infections with F. hepatica resulted in significantly higher metacercarial production than that noted with G. truncatula, in spite of a lower prevalence for the former lymnaeid. This finding allows for a significant decrease in the cost price of these larvae for commercial production.
Dar, Y; Vignoles, P; Rondelaud, D; Dreyfuss, G
Experimental infections of three Egyptian Pseudosuccinea columella populations with sympatric miracidia of Fasciola sp., coming from cattle- or sheep-collected eggs, were carried out to determine the capacity of this lymnaeid to support larval development of the parasite. Using microsatellite markers, the isolates of Egyptian miracidia were identified as Fasciola hepatica. Apart from being independent of snail origin, prevalences ranging from 60.4 to 75.5% in snails infected with five miracidia of F. hepatica were significantly higher than values of 30.4 to 42.2% in snails with bi-miracidial infections. The number of metacercariae ranged from 243 to 472 per cercarial-shedding snail and was independent of snail origin, parasite origin and miracidial dose used for infection. If P. columella was subjected to two successive bi-miracidial infections with F. hepatica, prevalence of infection was 63.3%, with a mean of 311 metacercariae per snail. These values were clearly greater than those already reported for Radix natalensis infected with the same parasite and the same protocol. Successful experimental infection of P. columella with F. hepatica suggests that this lymnaeid snail is an important intermediate host for the transmission of fascioliasis in Egypt.
Souza Cecília Pereira de
Full Text Available Fascioliasis is a parasitic disease of domestic ruminants that occurs worldwide. The lymnaeid intermediate hosts of Fasciola hepatica include Lymnaea columella, which is widely distributed in Brazil. A colony of L. columella from Belo Horizonte, MG, was reared in our laboratory to be used in studies of the F. hepatica life cycle, the intermediate host-parasite relationship and development of an anti-helminthic vaccine. In the first experiment 1,180 snails were exposed to miracidia of F. hepatica eggs removed from the biliary tracts of cattle from the State of Rio Grande do Sul. In the second and third experiments the snails were exposed to miracidia that had emerged from F. hepatica eggs from Uruguay, maintained in rabbits. The rates of infection in the first, second and third experiments were 0, 42.1 and 0% respectively. Over 15,806 metacercariae were obtained and stored at 4ºC. Four rabbits weighing 1.5 kg each were infected with 32-44 metacercariae and two with 200. Three rabbits begin to eliminate eggs of the parasite in the feces from 84 days after infection onwards. The biological cycle of F. hepatica in L. columella and the rabbit was completed within 124 days.
Liolope copulans (Trematoda: Digenea: Liolopidae) parasitic in Andrias japonicus (Amphibia: Caudata: Cryptobranchidae) in Japan: Life cycle and systematic position inferred from morphological and molecular evidence.
Baba, Takashi; Hosoi, Masatomi; Urabe, Misako; Shimazu, Takeshi; Tochimoto, Takeyoshi; Hasegawa, Hideo
The life cycle of Liolope copulans Cohn, 1902 (Trematoda: Digenea: Liolopidae), an intestinal parasite of the Japanese giant salamander Andrias japonicus (Temminck) (Amphibia: Caudata: Cryptobranchidae), was studied in the field and laboratory in Japan. This is the first description of mother sporocyst, daughter sporocyst and cercariae of a liolopid species. Non-oculate longifurcate pharyngeate cercariae were formed in lanceolate-cylindrical daughter sporocysts in Semisulcospira libertina (Gould) (Gastropoda: Sorbeoconcha: Pleuroceridae). They successfully developed to encapsulated metacercariae in cyprinid fishes, Nipponocypris sieboldii (Temminck and Schlegel) and Rhynchocypris lagowskii (Dybowski), by experimental infection. Cercariae had a V-shaped excretory vesicle with two looped arms, as in metacercariae and adults. Developmental stages from mother sporocyst to adult are described and illustrated. DNA sequencing was conducted for 28S and 18S rDNA of mother and daughter sporocysts, cercariae, and an adult. The result of molecular phylogenetic analysis suggests that L. copulans may be one of the basal taxa of the order Diplostomida Olson, Cribb, Tkach, Bray, and Littlewood, 2003, but its systematic position is still unclear because of the topological inconsistence between the 28S and 18S trees. Therefore, we tentatively place the family Liolopidae in the superfamily Diplostomoidea, mainly based on the morphology of sporocysts and cercariae. Copyright © 2011 Elsevier Ireland Ltd. All rights reserved.
Chai, Jong-Yil; Park, Young-Jin; Park, Jae-Hwan; Jung, Bong-Kwang
Mucosal immune responses against Pygidiopsis summa (Trematoda: Heterophyidae) infection were studied in ICR mice. Experimental groups consisted of group 1 (uninfected controls), group 2 (infection with 200 metacercariae), and group 3 (immunosuppression with Depo-Medrol and infection with 200 metacercariae). Worms were recovered in the small intestine at days 1, 3, 5, and 7 post-infection (PI). Intestinal intraepithelial lymphocytes (IEL), mast cells, and goblet cells were counted in intestinal tissue sections stained with Giemsa, astra-blue, and periodic acid-Schiff, respectively. Mucosal IgA levels were measured by ELISA. Expulsion of P. summa from the mouse intestine began to occur from days 3-5 PI which sustained until day 7 PI. The worm expulsion was positively correlated with proliferation of IEL, mast cells, goblet cells, and increase of IgA, although in the case of mast cells significant increase was seen only at day 7 PI. Immunosuppression suppressed all these immune effectors and inhibited worm reduction in the intestine until day 7 PI. The results suggested that various immune effectors which include IEL, goblet cells, mast cells, and IgA play roles in regulating the intestinal mucosal immunity of ICR mice against P. summa infection. PMID:24623878
Pinto, H A; Caffara, M; Fioravanti, M L; Melo, A L
Despite the large number of reports of species of Clinostomum from vertebrate hosts in South America, studies evaluating the molluscan transmitters of these parasites are scarce. In the present study, clinostomatoid cercariae shed from 0.02% (4/17,485) specimens of Biomphalaria spp., collected at the Pampulha reservoir, Belo Horizonte, Minas Gerais, Brazil, were used for experimental infection of Poecilia reticulata . Samples of cercariae from molluscs and metacercariae experimentally obtained from fish were subjected to morphological and molecular analyses and compared with species of Clinostomum reported in the Americas. The cercariae and metacercariae, here identified as Clinostomum sp., present general morphology similar to that reported for Clinostomum marginatum , however, from molecular point of view, differ significantly from North American C. marginatum and other species of Clinostomum reported in South America. These results suggest that the diversity of Clinostomum found in Brazil may be underestimated. Additional studies aimed at molecular characterization of South American species of Clinostomum, including the finding of specimens with sequences similar to that reported for C. marginatum in North America are required.
Effects of host species and life stage on the helminth communities of sympatric northern leopard frogs (Lithobates pipiens) and wood frogs (Lithobates sylvaticus) in the Sheyenne National Grasslands, North Dakota.
Gustafson, Kyle D; Newman, Robert A; Tkach, Vasyl V
We studied helminth communities in sympatric populations of leopard frogs (Lithobates pipiens) and wood frogs (Lithobates sylvaticus) and assessed the effects of host species and life stage on helminth community composition and helminth species richness. We examined 328 amphibians including 218 northern leopard frogs and 110 wood frogs collected between April and August of 2009 and 2010 in the Sheyenne National Grasslands of southeastern North Dakota. Echinostomatid metacercariae were the most common helminths found, with the highest prevalence in metamorphic wood frogs. Host species significantly influenced helminth community composition, and host life stage significantly influenced the component community composition of leopard frogs. In these sympatric populations, leopard frogs were common hosts for adult trematodes whereas wood frogs exhibited a higher prevalence of nematodes with direct life cycles. Metamorphic frogs were commonly infected with echinostomatid metacercariae and other larval trematodes whereas juvenile and adult frogs were most-frequently infected with directly transmitted nematodes and trophically transmitted trematodes. Accordingly, helminth species richness increased with the developmental life stage of the host.
Blasco-Costa, Isabel; Poulin, Robert; Presswell, Bronwen
Species of Apatemon Szidat, 1928 and Australapatemon Sudarikov, 1959 are reported from New Zealand for the first time, and their life cycles are resolved using molecular sequence data (28S and ITS rDNA regions and mitochondrial COI). The metacercaria of Apatemon sp. 'jamiesoni' ex Gobiomorphus cotidianus and its cercaria ex Potamopyrgus antipodarum are described in detail. Its adult, found in Anas platyrhynchos and Phalacrocorax punctatus, is identified by molecular sequence data. Apatemon sp. 'jamiesoni' uses a different species of snail host, exhibits consistent differences in the genetic markers examined and its single described adult differs from known species so as to be considered distinct, but its formal description awaits additional adult specimens. Australapatemon niewiadomski n. sp. is described from Anas platyrhynchos. It is distinguished morphologically by the absence of a ringnapf and its overall smaller size compared to most other Australapatemon spp. except Au. magnacetabulum and Au. minor, which are smaller in nearly all features than the new species. Au. niewiadomski n. sp. metacercaria and its intermediate host (Barbronia weberi) are identified via matching of molecular sequence data. The status of Apatemon and Australapatemon as distinct genera is confirmed based on their respective monophyly, and genetic divergence between them is comparable to other well-established genera in the Strigeidae. The diagnosis of Australapatemon is emended. Life history data, accurate patterns of host specialisation and distribution, alongside concurrent molecular and morphological evidence would be useful for an integrative taxonomical approach towards the elucidation of species diversity in this group.
Antunes, S.A.; Wiendl, F.M.; Almeida Dias, E.R.; Arthur, V.; Daniotti, C.
The mullet (Mugilidae) is a fish caught in large quantities in brackish and marine waters of the southern Brazilian coast, which in consequence of its raw consumption as ''sashimi'', a typical Japanese dish, has led to ten cases of human infection by Phagicola longa in Sao Paulo, Brazil. The mullet acts as a second intermediate host for the parasite, as in human heterophyiasis in the Near and Far East. In order to control these infections under commercial storage conditions and in raw consumption, the radiolysis of P. longa was studied in three mullet species - the silver mullet (Mugil curema), the grey mullet (M. platanus) and the ''paratipema'' (Mugil sp.) -subjected to ionization ranging from 1.0 to 10.0 kGy (2.21 kGy/h). It was observed that 1.0 and 2.0 kGy caused a motility decrease in silver mullet parasites from 100% to 15% and 17%, and that doses of 4.0 and 10.0 kGy caused metacercaria inviability. The parasite motility decreased in the grey mullet treated with doses of 2.0, 2.5, 3.0 and 3.5 kGy, from 56% to 31%, 9%, 18% and 5%, respectively, 4.0 kGy tending to be the control dose for P. Longa. This dose also controls other metacercaria found in the ''parati-pema'', without changing the odor, color or appearance of the treated mullet. (author)
Damborenea, C; Brusa, E; Paola, A
Pomacea canaliculata is a common gastropod in freshwater habitats from Central and Northern Argentina, extending northwards into the Amazon basin. Several Platyhelminthes have been reported associated to P. canaliculata, sharing an intimate relationship with this gastropod host. The objectives of this study were to describe the symbiotic species assemblages associated to P. canaliculata in the study area, and to disclose differences among them. Samples were taken in three typical small streams and one artificial lentic lagoon, all connected with the Rio de la Plata estuary. The 81.53% were infested with different symbiotic (sensu lato) species. Among the Platyhelminthes, the commensal Temnocephala iheringi Haswell, 1893 was highly prevalent in all samples, always in the mantle cavity. Four trematode taxa were recognized: (a) metacercariae of Echinostoma parcespinosum Lutz, 1924 in the mantle cavity and sporocysts in the digestive gland; (b) metacercariae of Dietziella egregia (Dietz, 1909) in the pericardial cavity; (c) unidentified xiphidiocercariae and (d) unidentified sporocysts and furcocercariae in the digestive gland. Nematode larvae and oligochaetes were found in two localities in the mantle cavity. Among the Annelida, Helobdella ampullariae Ringuelet, 1945 was found in the mantle cavity and lung of snails only from one locality. Our results show that although some of the symbionts are present in all localities, others are restricted to some particular ones, whether in their absolute numbers or in their relative abundance. Thus, each hosting population at the studied localities may be defined by the particular combination of symbionts that bears.
Jung, Younghun; Park, Yun-Kyu; Hwang, Myung-Gi; Soh, Chin-Thack
More than 1,500 clams of Corbicula fluminea, the most favorable food source of freshwater bivalves in Korea, were collected from 5 localities to examine cercarial and metacercarial infection with Echinostoma cinetorchis. Although 3 clams infected with suspicious E. cinetorchis metacercariae out of 200 specimens collected at Kangjin, Chollanam-do were detected, no cercarial and metacercarial infections with E. cinetorchis were observed in field-collected Corbicula specimens. In the susceptibility experiments with laboratory-reared clams, those infected with miracidia of E. cinetorchis did not release their cercariae up to 60 days after infection. To confirm the identity of second intermediate host of E. cinetorchis experimentally, a total of 30 clams were exposed to the cercariae from Segmentina hemisphaerula that had been infected with miracidia of E. cinetorchis. The clams were susceptible to cercariae of E. cinetorchis with an infection rate of 93.3%. Metacercariae from clams taken more than 7 days after cercarial exposure were fed to rats (S/D strain), and adult worms of E. cinetorchis, characterized by 37-38 collar spines on the head crown, were recovered from the ileocecal regions. This is the first report of C. fluminea as a possible second intermediate host of E. cinetorchis. PMID:11775336
Van, Chinh Dang; Doungchawee, Galayanee; Suttiprapa, Sutas; Arimatsu, Yuji; Kaewkes, Sasithorn; Sripa, Banchob
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. Copyright © 2016 Elsevier Ireland Ltd. All rights reserved.
Chai, Jong; Shin, Eun Hee; Guk, Sang Mee; Han, Eun Taek; Kim, Jae Lip [Seoul National University, Seoul (Korea)
To prevent some intestinal trematodes which are highly prevalent in Korea, the effect of irradiation on fishes and metacercariae has been analyzed for control of parasites. The metacercariae of intestinal trematodes were irradiated with various doses, and the infection rates and developmental status were assayed for estimation of optimal radiation doses. In addition, to examine the differences of susceptibility to radiation among the parasites, superoxide dismutase (SOD) activities were determined, and degree of apoptosis were observed. As a result, 100 Gy irradiation was sufficient for Metagonimus yokogawai to redue their infectivity and development, and 200 Gy was for Gymnophalloides seoi. While Neodiplostomum seoulense, a kind of intestinal rematode, was proved to be highly susceptible to radiation, sparganum and Anisakis were resistant to radiation. These might result from the differences of SOD activity among parasites. The apoptosis of parasites was mainly observed after 100 Gy irradiation, and further study was required to decide whether these phenomenon influences the susceptibility to radiation. 21 refs., 7 figs., 4 tabs. (Author)
Gonçalves, Raissa Alves; Oliveira, Marcos Sidney Brito; Neves, Ligia Rigôr; Tavares-Dias, Marcos
The present study investigated the effects of seasonal variation in parasites infracommunities of Hoplerythrinus unitaeniatus and Hoplias malabaricus from a tributary of Amazon River. For H. unitaeniatus and H. malabaricus, 11 parasite species were similar, and greatest parasite richness occurred during the rainy season. Ichthyophthirius multifiliis was the dominant parasite species for both hosts. In H. unitaeniatus, infection by Whittingtonocotyle caetei, Whittingtonocotyle jeju, Urocleidoides sp. and Anacanthorus sp. was higher during rainy season. Contracaecum sp., Procamallanus (Spirocamallanus) inopinatus, Nomimoscolex matogrossensis and Gorytocephalus spectabilis showed no seasonal pattern. For H. unitaeniatus, P. pillulare, Clinostomum marginatum and Argulus pestifer occurred only during dry season, while Trichodina sp., Dolops geayi, undetermined metacercariae and Posthodiplostomum sp. occurred only during the rainy season. In H. malabaricus, the prevalence of Urocleidoides eremitus was similar during the two seasons, but abundance was higher during the rainy season. Tetrahymena sp., C. marginatum, Dendrorchis neivai, undetermined metacercariae, Posthodiplostomum sp., Genarchella genarchella, Cystidicoloides sp., G. spectabilis, D. geayi, A. pestifer and Glossiphonidae gen. sp. occurred only during the dry season. However, Contracaecum sp. and P. (S.) inopinatus occurred during both seasons, but the prevalence of P. (S.) inopinatus was higher during the rainy season. Seasonal variation in this infection levels was due to the host's feeding behavior and habits and the availability of infectious forms of parasites with heteroxenic life cycles. The non-seasonal fluctuation detected are likely a result of the parasites biology, highly variable nature of this tributary of Amazon River and low abundance of parasites.
Aho, J.M.; Gibbons, J.W.; Esch, G.W.
The relationship between thermal loading and parasitism was examined in 980 mosquitofish, Gambusia affinis, taken from areas of varying thermal conditions at the Savannah River Plant (SRP) near Aiken, S. C. Collections were made at 2-week intervals from May to August 1974 and again from January to February 1975. The metacercaria of two strigeid trematodes, Ornithodiplostomum ptychocheilus and Diplostomum scheuringi were the only species of parasites recovered. Ornithodiplostomum ptychocheilus was always found encysted in the brain and eyes, whereas D. scheuringi was restricted exclusively to the body cavity. The density of the body-cavity parasite was highest in fish from areas of ambient temperatures and declined in fish from areas with higher water temperatures. The density of the brain parasite, on the other hand, was higher in fish from areas directly receiving thermal effluent than in fish from ambient-temperature areas. The body-cavity parasite was absent from mosquitofish from Pond C, but the infection percentages were relatively consistent in fish from other areas in the Par Pond system regardless of water temperature. The brain metacercaria were recovered from 95 percent of fish from Pond C
Joaber Pereira Junior
Full Text Available The orange disease is considered the main parasitosis in Perna perna mussel. It is caused by a complex life cycle bucephalid, involving three hosts, among them mollusks and fishes. With the aim of contributing to the knowledge of orange disease in mussel culture, the parasite life cycle was investigated. Experimental studies and monthly samples in the study area allowed the identification and characterization of the Bucephalus margaritae life cycle. Larvae and adults of B. margaritae were fixed in 5% formaldehyde, stained with Gomori’s trichrome, clarified in creosote and mounted in Canada balsam. The cercariae are found in the first intermediate host P. perna inside the sporocysts, which have the form of orange and ramified filaments. The metacercariae encysts in the gills and gill cavity of the second intermediate host, the blenniid Hypleurochilus fissicornis. The definitive host Menticirrhus americanus is infected ingesting blenniids parasitized with metacercariae. The high parasitological indexes of B. margaritae suggests that M. americanus and H. fissicornis act as the main definitive and intermediate hosts, respectively, in the trematode life cycle. The blenniid H. fissicornis is a new intermediate host to the species.
Carvallho, R P S; Takemoto, R M; Melo, C M; Jeraldo, V L S; Madi, R R
The catfish species Sciades proops inhabits muddy estuaries and shallow brackish lagoons, as well as freshwater. For these reasons, it is believed that this species may act as an intermediate, definitive and paratenic host in the life cycle of many parasites. From November 2010 to November 2011 and from August 2012 to July 2013, a total of 126 specimens of Sciades proops from the estuarine region of the Japaratuba River in the state of Sergipe, Brazil, were examined for parasites, of which 84.13% were infected by at least one species: Ergasilus sp. (Copepoda) (Prevalence P = 77.78%, Mean of Intensity MI = 10.08 ± 15.48, Mean Abundance MA = 14.27 ± 7.48) in the gills, Contracaecum sp. (P = 23.02%, MI = 20.59 ± 80.58, MA =39.12 ± 4.47) in the general cavity, Procamallanus sp. (P = 0.79%, MI = 1, MA = 0.01 ± 0.09), Raphidascaroides sp. (P = 2.31%, MI = 1.33 ± 0.58, MA = 0.33 ± 0.22) and Cuccullanus sp. (Nematoda) (P = 0.79%, MI = 1, MA = 0.01 ± 0.09) in the intestine, Ancyrocephalinae (Monogenea) (P = 0.79%, MI = 1, MA = 0.01 ± 0.09) in the gills, Pseudoacanthostomumpanamense (P = 1.59%, MI = 9.00 ± 8.49, MA = 0.14 ± 1.36) in the intestine, Clinostomum sp. (P = 1.59%, MI = 17.50 ± 23.33, MA = 0.29 ± 3.03) on the body surface and two unidentified metacercariae, referred to as Metacercaria 1 (P = 0.79%; IM = 1, MA = 0.01 ± 0.09) and Metacercaria 2 (Digenea) (P = 0.79%, MI = 7, MA = 0.06 ± 0.62) in the swim bladder. Ergasilus sp. was the dominant species and thus classified as core, with Contracaecum sp. as the satellite and other species as secondary species. The spatial distribution of infection with Ergasilus sp. and Contracaecum sp. showed a typical pattern of aggregate distribution. The sex of the host did not influence parasitic infections, but infection with Ergasilus sp. showed a positive and significant correlation with biometric and epidemiologic parameters, whereas infection with Contracaecum sp. was correlated only with prevalence and abundance.
R. P. S. Carvallho
Full Text Available Abstract The catfish species Sciades proops inhabits muddy estuaries and shallow brackish lagoons, as well as freshwater. For these reasons, it is believed that this species may act as an intermediate, definitive and paratenic host in the life cycle of many parasites. From November 2010 to November 2011 and from August 2012 to July 2013, a total of 126 specimens of Sciades proops from the estuarine region of the Japaratuba River in the state of Sergipe, Brazil, were examined for parasites, of which 84.13% were infected by at least one species: Ergasilus sp. (Copepoda (Prevalence P = 77.78%, Mean of Intensity MI = 10.08 ± 15.48, Mean Abundance MA = 14.27 ± 7.48 in the gills, Contracaecum sp. (P = 23.02%, MI = 20.59 ± 80.58, MA =39.12 ± 4.47 in the general cavity, Procamallanus sp. (P = 0.79%, MI = 1, MA = 0.01 ± 0.09, Raphidascaroides sp. (P = 2.31%, MI = 1.33 ± 0.58, MA = 0.33 ± 0.22 and Cuccullanus sp. (Nematoda (P = 0.79%, MI = 1, MA = 0.01 ± 0.09 in the intestine, Ancyrocephalinae (Monogenea (P = 0.79%, MI = 1, MA = 0.01 ± 0.09 in the gills, Pseudoacanthostomumpanamense (P = 1.59%, MI = 9.00 ± 8.49, MA = 0.14 ± 1.36 in the intestine, Clinostomum sp. (P = 1.59%, MI = 17.50 ± 23.33, MA = 0.29 ± 3.03 on the body surface and two unidentified metacercariae, referred to as Metacercaria 1 (P = 0.79%; IM = 1, MA = 0.01 ± 0.09 and Metacercaria 2 (Digenea (P = 0.79%, MI = 7, MA = 0.06 ± 0.62 in the swim bladder. Ergasilus sp. was the dominant species and thus classified as core, with Contracaecum sp. as the satellite and other species as secondary species. The spatial distribution of infection with Ergasilus sp. and Contracaecum sp. showed a typical pattern of aggregate distribution. The sex of the host did not influence parasitic infections, but infection with Ergasilus sp. showed a positive and significant correlation with biometric and epidemiologic parameters, whereas infection with Contracaecum sp. was correlated only with
Rojas-Caraballo, Jose; López-Abán, Julio; Fernández-Soto, Pedro; Vicente, Belén; Collía, Francisco; Muro, Antonio
Fasciolosis is listed as one of the most important neglected tropical diseases according with the World Health Organization and is also considered as a reemerging disease in the human beings. Despite there are several studies describing the immune response induced by Fasciola hepatica in the mammalian host, investigations aimed at identifying the expression profile of genes involved in inducing hepatic injury are currently scarce. Data presented here belong to a time-course investigation of the gene expression profile in the liver of BALB/c mice infected with F. hepatica metacercariae at 7 and 21 days after experimental infection. The data published here have been deposited in NCBI's Gene Expression Omnibus and are accessible through GEO Series accession number GSE69588, previously published by Rojas-Caraballo et al. (2015) in PLoS One .
Pérez, J; Ortega, J; Moreno, T; Morrondo, P; López-Sández, C; Martínez-Moreno, A
Hepatic lesions and changes in the hepatic lymph nodes were examined in groups of sheep infected, and sometimes reinfected, with metacercariae of Fasciola hepatica, with or without treatment with triclabendazole (TCBZ). Hepatic damage was more severe in sheep given small repeated ("trickle") infections than in those given two larger, spaced doses. Inflammatory infiltrates of eosinophils, CD3+ T cells, CD79 alpha+ B cells and IgG+ plasma cells, and the presence of granulomata, were particularly marked in trickle-infected groups, suggesting that the host response to certain fluke or egg antigens causes severe hepatic damage. TCBZ administration induced the elimination of flukes and healing of the majority of hepatic lesions but did not prevent severe hepatic damage produced by later infections.
DA Silva, Aleksandro S; Baldissera, Matheus D; Bottari, Nathieli B; Gabriel, Matheus E; Rhoden, Leandro A; Piva, Manoela M; Christ, Ricardo; Stedille, Fernanda A; Gris, Anderson; Morsch, Vera M; Schetinger, Maria Rosa; Mendes, Ricardo E
The aim of this study was to evaluate the oxidative stress in serum and liver and adenosine deaminase (ADA) activity of cattle experimentally infected by Fasciola hepatica. The group A consisted of five healthy animals (uninfected), and the group B was composed of five animals orally infected with 200 metacercariae of F. hepatica. On days 20, 40, 60 and 80 post-infection (PI) serum was collected to measure oxidative stress variables. On day 100 PI, animals were humanely euthanized and liver samples were collected. Infected animals showed lower (P hepatic reactive oxygen species (ROS) were higher (P Hepatic thiobarbituric acid reactive substances were higher (P hepatic damage, and that increased in hepatic ADA activity may contribute to the inflammatory process.
Fischer, Peter U; Curtis, Kurt C; Marcos, Luis A; Weil, Gary J
Human paragonimiasis is an emerging disease in Missouri. To characterize local parasites, we examined crayfish from three rivers. Metacercaeriae consistent with Paragonimus kellicotti were detected in 69%, 67%, and 37% of crayfish from the Big Piney, Huzzah, and Black Rivers, respectively. Sequencing of the second internal transcribed spacer and other DNA markers confirmed the species identification and the presence of identical parasite sequences in clinical specimens from two human cases. Mongolian gerbils were infected by intraperitoneal injection with 3-8 metacercariae. Most gerbils died 15-49 days post-infection. Necropsies showed pulmonary hemorrhage with necrosis, and flukes as long as 8 mm were recovered from intrathoracic tissues. Western blot analysis using P. kellicotti antigen showed a strong antibody response in gerbils 39 days post-infection. These results demonstrate that P. kellicotti is common in Missouri crayfish. The gerbil model may be useful for research on the pathogenesis, immunology, and treatment of paragonimiasis.
Nissen, Sofie; Nguyen, Lan Anh; Thamsborg, Stig Milan
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...... technique, temperature and weight of the dogs were measured as was total white blood cells, eosinophils and microhaemotocrit values. Subsets of dogs were examined post-mortem for presence of adult FZT at three different time points post infection. Patent infections established in all eight infected dogs...
Full Text Available Fasciolosis is listed as one of the most important neglected tropical diseases according with the World Health Organization and is also considered as a reemerging disease in the human beings. Despite there are several studies describing the immune response induced by Fasciola hepatica in the mammalian host, investigations aimed at identifying the expression profile of genes involved in inducing hepatic injury are currently scarce. Data presented here belong to a time-course investigation of the gene expression profile in the liver of BALB/c mice infected with F. hepatica metacercariae at 7 and 21 days after experimental infection. The data published here have been deposited in NCBI's Gene Expression Omnibus and are accessible through GEO Series accession number GSE69588, previously published by Rojas-Caraballo et al. (2015 in PLoS One .
Ocorrência de infecção natural de Fasciola hepatica Linnaeus, 1758 em Lymnaea columella Say, 1817 no Vale do Paraíba, SP, Brasil Natural infection by Fasciola hepatica in Lymnaea columella in the Paraíba river valley, S. Paulo, Brazil
Marlene Tiduko Ueta
Full Text Available Foram registradas em Piquete, no vale do rio Paraíba do Sul (SP, Brasil, taxas de 1,22% e 0,14% de infecção natural em Lymnaea columella, por Fasciola hepatica. Em um único exemplar de Lymnaea columella dentre os 1.052 examinados, foram observadas rédias com xifidiocercárias, rédias com cercárias de Fasciola hepatica e metacercárias de Echinostomatidae.Infection rates of 1.22% and 0.14% were obtained in Lymnaea columella snails naturally infected by Fasciola hepatica. Samples of the snails were collected in Piquete, a municipality of Paraíba do Sul, a river valley area in the State of S. Paulo. Also observed was one of the 1052 specimen of the Lymnaea columella rediae which had xiphidiocercariae and rediae with Fasciola hepatica cercariae and metacercariae of Echinostomatidae.
Pinto, Hudson Alves; de Melo, Alan Lane
Eurytremiasis is an important parasitic disease of cattle that was recently suggested to be a neglected and emerging human disease in Brazil. Based on a misinterpretation of the life cycle of the parasite, it was suggested that a great number of people could be infected with this fluke in the country. In the present letter, aspects of the life cycle of Eurytrema spp. are revisited and clarified. The mechanism of transmission previously reported for the few accidental human cases involved the ingestion of raw or undercooked insects (grasshoppers and crickets) harboring the infective metacercariae. In reality, the zoonotic potential of Eurytrema species is extremely low, and human eurytremiasis is not, and probably never will be, a zoonotic disease in countries where entomophagy is not a common food habit.
Full Text Available Fascioliasis is a worldwide but unevenly distributed zoonosis caused by the trematode Fasciola hepatica that infects domesticated herbivores. Fasciolosis also occurs accidentally in humans by ingestion of metacercaria-laden freshwater or water plants. Human infections are common in developing countries and are not rare in Europe. The clinical course has been conventionally described in two phases: an acute phase of hepatic parenchymal invasion of an immature worm larva (parenchymal phase and a stationary phase after residence in the bile duct and production of eggs (ductal phase. We report a 34 years old woman from Ilam, western Iran with hepatic disorder, RUQ pain, and jaundice. The diagnosis was made by sonography, CT scan and serologic studies. Serologic exam (ELISA was positive & CT findings were compatible with fascioliasis.
Sohn, Woon-Mok; Seo, Min
The surface ultrastructure of Acanthotrema felis (Trematoda: Heterophyidae) adults, recovered from a kitten experimentally infected with the metacercariae, was observed using a scanning electron microscope. The worm was leaf-like, ventrally concave and covered with scale-like multi-pointed tegumental spines. The spines on the anterior surface were short but broad, and had 10-12 pointed tips. The cytoplasmic processes protruded around the spines, like pockets for the spines. The ventrogenital opening was crescent, or kidney-shaped, and had protuberances with minute spines on its surrounding tegument. The spines on the posterior surface were long, but narrow, with 6-8 pointed tips. The cytoplasmic processes on this tegument were ridge-like, and elevated along the row of the spines. The surface ultrastructure of A. felis is generally similar to that of other heterophyid flukes, but some features are characteristic, and may be of taxonomic and bio-ecological significance. PMID:12815322
Ataev, G L; Tokmakova, A S
Infection of molluscs Succinea putris by trematodes Leucochloridium paradoxum was studied in the region of Vyritsa (Leningrad Province) during the period of 2008-2014. On the basis of the obtained data, seasonal dynamics of infection of molluscs can be presented as follows. Infection of S. putris occurs during the whole warm period from May to August. Young sporocysts of L. paradoxum overwinter and the metacercariae that develop in their extensions mature during spring becoming infective for birds. In the second half of summer, sporocysts start degenerating and die in late August-September. Each sporocyst can form 2-3 mature broodsacs (maximum 5) simultaneously. In cases of multiple infections, their number can reach 19. Several cases of independent release of sporocysts from molluscs were observed. They survive in environment for about an hour, retaining the ability to infect definitive hosts. Additionally, birds can be infected by pecking of horns of infected snails.
Ma. del Carmen Guzmán-Cornejo
Full Text Available In September, 1995 and January, 1996, six species of fishes from Lake Cuitzeo, Michoacan, Mexico, were collected (Alloophorus robustus n=30, Goodea atripinnis n=30, Xenotoca variata n=41, Chirostoma jordani n=30, Carassius auratus n=30 and Oreochromis aureus n=30, to establish the infection levels reached by Posthodiplostomum minimum, Clinostomum complanatum and Diplostomum (Tylodelphys sp., in their hosts. Infection caused by P. minimum metacercariae was the most important helminthiasis considering the number of hosts they parasite (five of six, and the high levels of abundance of the infection; we suggest that fish preferences and the generalist character of the trematode are the causes that determine this infection. Finally, the role of goodeids and atherinids in transmission of infections in the lake is confirmed; the former being the main group mantaining such infections.
Chikhliaeva, I V; Kirillov, A A; Kirillova, N Iu
Data on trematodes of the order Plagiorchiida from 11 species of amphibians from the Middle Volga region are given. Literary data of the last 30 years, combined with the results of our own research are presented together for the first time. Finds of 17 trematode species are reliable. From them, metacercaria of Astiotrema monticelli is recorded from amphibians of the fauna of Russia for the first time; Dolichosaccus rastellus and Paralepoderma cloacicola are recorded for the first time from amphibians of the Middle Volga region. New hosts are mentioned for 8 species of parasites. Trematodes of edible frog Rana esculenta Linnaeus, 1758, a new species to the region, are described for the first time. The following characteristics are given for each trematode species: its taxonomic position, set of hosts, localization, places of detection, biology, geographical distribution, and the degree of host specificity. Morphological descriptions and original figures are given for 12 trematode species.
Fernandes, Berenice Maria Musco; Justo, Marcia Cristina Nascimento; Anjos, Camila Saraiva Dos; Malta, José Celso de Oliveira; Dumbo, José Chissiua
This study presents the helminthofauna of digenean parasites of Acestrorhynchus falcirostris in the state of Amazonas (Brazil). Eight species belonging to four families were recovered: Metacercariae of Austrodiplostomum compactum (Diplostomidae) and Clinostomum marginatum (Clinostomidae) and mature specimens of Bellumcorpus majus, Prosorhynchoides carvajali, Rhipidocotyle santanaensis (Bucephalidae); Prosthenhystera obesa (Callodistomidae); Genarchella genarchella (Derogenidae) and Phyllodistomum spatula (Gorgoderidae). Examining specimens belonging to the genus Bellumcorpus collected from A. falcirostris, were observed a great variation in the size and shape of testes, which also could be observed in the type specimens of Bellumcorpus majus and B. schubarti. Considering the similar morphology and morphometric characteristics of both species, in this paper B. schubarti is considered synonym of B. majus. Acestrorhynchus falcirostris is a new host records for all these species of digenean, except to A. compactum and C. marginatum. Prosorhynchoides carvajali is referred for the first time in Brazil.
Berenice Maria Musco Fernandes
Full Text Available Abstract This study presents the helminthofauna of digenean parasites of Acestrorhynchus falcirostris in the state of Amazonas (Brazil. Eight species belonging to four families were recovered: Metacercariae of Austrodiplostomum compactum (Diplostomidae and Clinostomum marginatum (Clinostomidae and mature specimens of Bellumcorpus majus, Prosorhynchoides carvajali, Rhipidocotyle santanaensis (Bucephalidae; Prosthenhystera obesa (Callodistomidae; Genarchella genarchella (Derogenidae and Phyllodistomum spatula (Gorgoderidae. Examining specimens belonging to the genus Bellumcorpus collected from A. falcirostris, were observed a great variation in the size and shape of testes, which also could be observed in the type specimens of Bellumcorpus majus and B. schubarti. Considering the similar morphology and morphometric characteristics of both species, in this paper B. schubarti is considered synonym of B. majus. Acestrorhynchus falcirostris is a new host records for all these species of digenean, except to A. compactum and C. marginatum. Prosorhynchoides carvajali is referred for the first time in Brazil.
The Apple Snail Pomacea maculata (Caenogastropoda: Ampullariidae) as the intermediate host of Stomylotrema gratiosus (Trematoda: Stomylotrematidae) in Brazil: the first report of a mollusc host of a Stomylotrematid Trematode.
Pinto, Hudson A; Cantanhede, Selma Patrícia D; Thiengo, Silvana C; de Melo, Alan L; Fernandez, Monica A
Trematodes belonging to the family Stomylotrematidae are intestinal parasites of birds. Despite the worldwide distribution and diversity of host species, the first intermediate host remains unknown. For a survey of parasites of Pomacea maculata , snails were collected from the municipality of São Vicente Férrer, state of Maranhão, northeastern Brazil. In the present study, the xiphidiocercariae shed from these snails were used in the experimental infection of the water bug Belostoma plebejum. The insect mortality was observed 30 days post-infection, and the metacercariae recovered in the body cavity of B. plebejum were identified as Stomylotrema gratiosus. This is the first report of an ampullariid snail as intermediate host of stomylotrematid trematodes.
Pistl, J; Mikula, I; Krupicer, I; Snirc, J
The effect of oral dosing with emissions containing heavy metals (mercury, lead, copper, cadmium, zinc and chromium) on immune status was tested in 3 groups of sheep. Two groups (Groups I and II) were given emissions for 15 d. One of these groups (Group I) was then also infested with Fasciola hepatica metacercaria on day 16. These 2 groups (Groups I and II) and a control group (Group C) were immunized with a Listeria vaccine on days 8 and 22. A decreased index of metabolic response (IMR) of phagocytes and reduced responses of lymphocytes to mitogenic activation with phytohemagglutinin and L monocytogenes antigen stimulation in the leukocyte migration-inhibition test were recorded. Decreased agglutination titers of serum antibodies against L monocytogenes were observed. The F hepatica infestation had no significant effect on the migration index or IMR.
Fredensborg, Brian Lund; Poulin, R
Density-dependent effects on parasite fitness have been documented from adult helminths in their definitive hosts. There have, however, been no studies on the cost of sharing an intermediate host with other parasites in terms of reduced adult parasite fecundity. Even if larval parasites suffer...... transmission to their bird definitive host by predation. In experimental infections, we found an intensity-dependent establishment success, with a decrease in the success rate of cercariae developing into infective metacercariae with an increasing dose of cercariae applied to each amphipod. In natural...... the two species. Our results thus indicate that the infracommunity of larval helminths in their intermediate host is interactive and that any density-dependent effect in the intermediate host may have lasting effects on individual parasite fitness....
Latham, A D M; Fredensborg, Brian Lund; McFarland, L H
The whelk Cominella glandiformis is an important predator-scavenger of New Zealand intertidal ecosystems; a few whelks can quickly eat all the soft tissues of recently dead crabs. In this study, we demonstrate that whelks can also ingest and act as paratenic hosts for at least 4 helminth species...... of parasites during their short stay in the whelks is not affected by whelk size. The majority of metacercariae and nematodes are passed out in whelk feces within 3 days of ingestion, whereas the few cystacanths found did not leave whelks until after that time; no parasite was left in whelks 5 days...... postingestion. Survival of all 4 helminth species was generally very high, though it decreased day by day in 2 species. Given that the avian definitive hosts of all 4 helminths also eat whelks, our results indicate that alternative transmission pathways exist and that parasites can take routes through food webs...
Full Text Available Eleven species of trematode larvae (sporocysts and cercariae of Bucephalus labracis Paggi et Orecchia, 1965 and Cercaria lata Lespes, 1857 and metacercariae of Acanthoparyphium sp. Dietz , 1909, Caecincola parvulus Marshall et Gilbert, 1905, Curtuteria australis Allison, 1979, Gymnophallus fossarum Bartoli, 1965, Gymnophallus rebecqui Bartoli, 1983, Lepocreadium pegorchis Stossich, 1901, Parazoogonus sp. Looss, 1901, Psilostomum brevicolle Creplin, 1829 and Robphildollfusium fractum Rudolphi, 1819 were found in the clam (Ruditapes decussatus from Tunisian coasts. A part from B. labracis and Cercaria lata, the other trematode species were recorded for the first time in Tunisia. The examination of the frequency of recorded parasites reveals some monthly variation. This variation is statistically significant by comparing the sampling sites. These results indicate that more frequent parasites (G. fossarum, Cercaria lata, Curtuteria australis and Acanthoparyphium sp. could be considered as biological indicators making it possible to predict the capture localities of the clams.
Shyu, L Y; Lee, H H; Chen, E R
Stool samples of 817 children (grades 4-6) from 3 primary schools, were collected and examined by the antiformin-ether concentration technique. Results showed no cases of Fasciolopsis were found from Liu-Ying primary school (Liu-Ying Hsiang), but infection rates of this fluke were found to be 13.0% and 3.5% for Ho-Chien and Tung-Hsing primary schools (Hsia-Ying Hsiang) respectively. Fourteen infected children had recent experience of eating raw water caltrops. On the other hand, only 8(21.6%) of 37 non-infected children had eaten raw water caltrops. Examination of intermediate host (Segmentina hemisphaerula, Trapa bispinosa) results showed that neither cercaria nor metacercaria was found. Intestines and stool samples from pigs were examined for worms and eggs of Fasciolopsis, the results were also negative.
Full Text Available BACKGROUND: Clonorchiasis, caused by the infection of Clonorchis sinensis (C. sinensis, is a kind of neglected tropical disease, but it is highly related to cholangiocarcinoma and hepatocellular carcinoma (HCC. It has been well known that the excretory/secretory products of C. sinensis (CsESPs play key roles in clonorchiasis associated carcinoma. From genome and transcriptome of C. sinensis, we identified one component of CsESPs, severin (Csseverin, which had three putative gelsolin domains. Its homologues are supposed to play a vital role in apoptosis resistance of tumour cell. METHODOLOGY/PRINCIPAL FINDINGS: There was significant similarity in tertiary structures between human gelsolin and Csseverin by bioinformatics analysis. We identified that Csseverin expressed at life stage of adult worm, metacercaria and egg by the method of quantitative real-time PCR and western blotting. Csseverin distributed in vitellarium and intrauterine eggs of adult worm and tegument of metacercaria by immunofluorence assay. We obtained recombinant Csseverin (rCsseverin and confirmed that rCsseverin could bind with calciumion in circular dichroism spectrum analysis. It was demonstrated that rCsseverin was of the capability of actin binding by gel overlay assay and immunocytochemistry. Both Annexin V/PI assay and mitochondrial membrane potential assay of human hepatocarcinoma cell line PLC showed apoptosis resistance after incubation with different concentrations of rCsseverin. Morphological analysis, apoptosis-associated changes of mitochondrial membrane potential and Annexin V/PI apoptosis assay showed that co-incubation of PLC cells with rCsseverin in vitro led to an inhibition of apoptosis induced by serum-starved for 24 h. CONCLUSIONS/SIGNIFICANCE: Collectively, the molecular properties of Csseverin, a molecule of CsESPs, were characterized in our study. rCsseverin could cause obvious apoptotic inhibition in human HCC cell line. Csseverin might exacerbate the
Villa-Mancera, Abel; Méndez-Mendoza, Maximino
Fluke burdens, liver fluke size and biomass, fecal eggs counts, serum levels of hepatic enzymes and immune response were assessed in sheep immunized with peptide mimotopes of cathepsin L1 and challenged with Fasciola hepatica metacercariae. Twenty sheep were randomly allocated to four groups of five animals each; groups 1 and 2 were immunized at weeks 0 and 2 with mimotopes YVYRWVEAECVA and FSPAYLRDAALK, respectively; group 3 was immunized with wild-type M13KE phage and the control group received phosphate-buffered saline (PBS). All groups were challenged with 300 metacercariae at week 6 and slaughtered 16 weeks later. Groups 1 and 2 showed a reduction in fluke burden of 51.7% and 35.9%, respectively, when compared to the control group, but only the former was significant at the 5% level. Vaccinated animals showed a significant reduction in fluke length and width, wet weights and egg output. A significant diminution in the total biomass of parasites recovered was also observed in group 1. Levels of anti-phage total IgG increased rapidly within 2 weeks of the first immunization and were always significantly higher in groups 1 and 2 than in the infected control group. The fluke burden in group 1 was significantly correlated with IgG1 and total IgG. The vaccinated sheep with phage clones produced significantly high titres of IgG1 and IgG2 antibodies indicating a mixed Th1/Th2 response. Copyright © 2012 Elsevier Ltd. All rights reserved.
Michael E Colvin
Full Text Available Most pathogen detection tests are imperfect, with a sensitivity < 100%, thereby resulting in the potential for a false negative, where a pathogen is present but not detected. False negatives in a sample inflate the number of non-detections, negatively biasing estimates of pathogen prevalence. Histological examination of tissues as a diagnostic test can be advantageous as multiple pathogens can be examined and providing important information on associated pathological changes to the host. However, it is usually less sensitive than molecular or microbiological tests for specific pathogens. Our study objectives were to 1 develop a hierarchical occupancy model to examine pathogen prevalence in spring Chinook salmon Oncorhynchus tshawytscha and their distribution among host tissues 2 use the model to estimate pathogen-specific test sensitivities and infection rates, and 3 illustrate the effect of using replicate within host sampling on sample sizes required to detect a pathogen. We examined histological sections of replicate tissue samples from spring Chinook salmon O. tshawytscha collected after spawning for common pathogens seen in this population: Apophallus/echinostome metacercariae, Parvicapsula minibicornis, Nanophyetus salmincola/ metacercariae, and Renibacterium salmoninarum. A hierarchical occupancy model was developed to estimate pathogen and tissue-specific test sensitivities and unbiased estimation of host- and organ-level infection rates. Model estimated sensitivities and host- and organ-level infections rates varied among pathogens and model estimated infection rate was higher than prevalence unadjusted for test sensitivity, confirming that prevalence unadjusted for test sensitivity was negatively biased. The modeling approach provided an analytical approach for using hierarchically structured pathogen detection data from lower sensitivity diagnostic tests, such as histology, to obtain unbiased pathogen prevalence estimates with
Cheng, You-zhu; Lin, Guo-hua; Li, You-song
To investigate the natural resources of the freshwater crab which can serve as the intermediate hosts of Paragonimus in Pinghe and Youxi of Fujian Province. Freshwater crabs were collected. The morphological characteristics of the crabs and the habitats were observed. The crabs were dissected and examined for the presence of Paragonimus metacercariae. Two new species of crabs were described, named as Sinopotamon zhangzhouense sp. nov. and Bottapotamon youxiense sp. nov.. S. zhangzhouense sp. nov., holotype: male, carapace length 35.9 mm, breadth 42.8 mm, thickness 18.6 mm, collected from Pinghe County in southwest of Fujian. (24 degrees 14.206' N, 117 degrees 12.594' E). Distal segment of the first pleopod of male tended flattish, and showed palm nest-shaped concave, which divided into two point leafs, and longitudinal crack clearly identified in back. The end half of distal segment reversed to ventral outwardly. This species usually lived in the sluggish stream. The infection rate of Paragonimus westermani and P. cenocopiosus in S. zhangzhouense sp. nov. was 44.9% (35/78). B. youxiense sp. nov., holotype: male, carapace length 13.35 mm, breadth 16.63 mm, thickness 7.20 mm, collected from Youxi County in central Fujian (26 degrees 10.558' N, 118 degrees 22.012' E). The first pleopod of male was in slightly flat shape, ample and developed, as bow-like uplift. This species usually lived in the relatively flat terrain of stream. The infection rate of P. skrjabini metacercariae in B. youxiense sp. nov. was 92.1% (58/63). Two new species of freshwater crabs (S. zhangzhouense sp nov. and Ryouxiense sp. nov.) serving as the intermediate hosts of Paragonimus have been described.
Full Text Available Purpose: To determine the prevalence of paragonimiasis among the patients who were attending the tuberculosis (TB clinics at the Community Health Centre, Pfutsero, Phek District, Nagaland. To determine the species of Paragonimus that cause infection in humans and the crustacean host that acts as the infectious source for humans. Materials and Methods: Sputum specimens were examined microscopically for Paragonimus eggs and acid fast bacilli. Blood samples were tested by microenzyme-linked immunosorbant assay for Paragonimus-specific immunoglobulin G antibodies. Crab extracts prepared by digestion with artificial gastric juice were examined for Paragonimus metacercariae under a stereoscopic microscope. The species identification of the parasite was based on morphological and molecular characterizations of eggs and metacercariae employing polymerase chain reaction and DNA sequencing. Results: Seven out of the 14 patients tested seropositive for paragonimiasis and Paragonimus eggs were detected in sputum of two out of the seven seropositive patients, indicating a prevalence of 50% and an egg detection rate of 14%, respectively. The prevalence was highest in the 10-30 year age group. More males got the infection than females, the ratio being 5:2. P. heterotremus was identified as the causative agent of human paragonimiasis and Potamiscus manipurensis as the crab host. Conclusions: The study revealed that paragonimiasis has been endemic in Pfutsero, Nagaland, and half of the patients attending the TB clinic were actually suffering from pulmonary paragonimiasis. This is the first confirmed report of an endemic focus of paragonimasis and description of P. heterotremus as the causative agent in Nagaland, India.
Oral delivery of Bacillus subtilis spores expressing cysteine protease of Clonorchis sinensis to grass carp (Ctenopharyngodon idellus): Induces immune responses and has no damage on liver and intestine function.
Tang, Zeli; Sun, Hengchang; Chen, TingJin; Lin, Zhipeng; Jiang, Hongye; Zhou, Xinyi; Shi, Cunbin; Pan, Houjun; Chang, Ouqin; Ren, Pengli; Yu, Jinyun; Li, Xuerong; Xu, Jin; Huang, Yan; Yu, Xinbing
Clonorchis sinensis (C. sinensis) is a fish-borne trematode. Human can be infected by ingestion of C. sinensis metacercariae parasitized in grass carp (Ctenopharyngodon idella). For induction of effective oral immune responses, spores of Bacillus subtilis (B. subtilis) WB600 were utilized as vehicle to delivery CsCP (cysteine protease of C. sinensis) cooperated with CotC (B.s-CotC-CP), one of coat proteins, to the gastrointestinal tract. After routine culture of 8-12 h in LB medium, B. subtilis containing CotC-CsCP was transferred into the sporulation culture medium. SDS-PAGE, western blotting and the growth curve indicated that the best sporulation time of recombinant WB600 was 24-30 h at 37 °C with continuous shaking (250 rpm). Grass carp were fed with three levels of B.s-CotC-CP (1 × 10 6 , 1 × 10 7 , and 1 × 10 8 CFU g -1 ) incorporated in the basal pellets diet. The commercial pellets or supplemented with spores just expressing CotC (1 × 10 7 CFU g -1 ) were served as control diet. Our results showed that grass carp orally immunized with the feed-based B.s-CotC-CP developed a strong specific immune response with significantly (P sinensis in fish body. Therefore, this study demonstrated that the feed-based recombinant spores could trigger high levels of mucosal and humoral immunity, and would be a promising candidate vaccine against C. sinensis metacercariae formation in freshwater fish. Copyright © 2017 Elsevier Ltd. All rights reserved.
Chen, Tingjin; Jiang, Hongye; Sun, Hengchang; Xie, Zhizhi; Ren, Pengli; Zhao, Lu; Dong, Huimin; Shi, Mengchen; Lv, Zhiyue; Wu, Zhongdao; Li, Xuerong; Yu, Xinbing; Huang, Yan; Xu, Jin
Clonorchis sinensis, the causative agent of clonorchiasis, is classified as one of the most neglected tropical diseases and affects more than 15 million people globally. This hepatobiliary disease is highly associated with cholangiocarcinoma. As key molecules in the infectivity and subsistence of trematodes, glycolytic enzymes have been targets for drug and vaccine development. Clonorchis sinensis pyruvate kinase (CsPK), a crucial glycolytic enzyme, was characterized in this research. Differences were observed in the sequences and spatial structures of CsPK and PKs from humans, rats, mice and rabbits. CsPK possessed a characteristic active site signature (IKLIAKIENHEGV) and some unique sites but lacked the N-terminal domain. The predicted subunit molecular mass (Mr) of CsPK was 53.1 kDa. Recombinant CsPK (rCsPK) was a homopentamer with a Mr. of approximately 290 kDa by both native PAGE and gel filtration chromatography. Significant differences in the protein and mRNA levels of CsPK were observed among four life stages of C. sinensis (egg, adult worm, excysted metacercaria and metacercaria), suggesting that these developmental stages may be associated with diverse energy demands. CsPK was widely distributed in adult worms. Moreover, an intense Th1-biased immune response was persistently elicited in rats immunized with rCsPK. Also, rat anti-rCsPK sera suppressed C. sinensis adult subsistence both in vivo and in vitro. The sequences and spatial structures, molecular mass, and expression profile of CsPK have been characterized. rCsPK was indicated to be a homopentamer. Rat anti-rCsPK sera suppressed C. sinensis adult subsistence both in vivo and in vitro. CsPK is worthy of further study as a promising target for drug and vaccine development.
Rojas-Caraballo, Jose; López-Abán, Julio; Fernández-Soto, Pedro; Vicente, Belén; Collía, Francisco; Muro, Antonio
Fasciola hepatica infection still remains one of the helminthic neglected tropical diseases (NTDs). It has a huge worldwide distribution, affecting mainly cattle and, sometimes, human beings. In addition to data reported about the immunological response induced by helminthic infections and that induced by Fasciola hepatica, little is known about the gene expression profile in its organ target, the liver, which is where adult worms are established and live for long periods of time, causing its characteristic pathology. In the present work, we study both the early and late gene expression profiles in the livers of mice infected with F. hepatica metacercariae using a microarray-based methodology. A total of 9 female-6-week-old BALB/c mice (Charles River Laboratories, Barcelona, Spain) weighing 20 to 35 g were used for the experiments. Two groups of BALB/c mice were orally infected with seven F. hepatica metacercariae, and the other group remained untreated and served as a control. Mice were humanely euthanized and necropsied for liver recovery, histological assessment of hepatic damage, RNA isolation, microarray design and gene expression analysis on the day of infection (t0), seven days post-infection (t7) and twenty-one days post-infection (t21). We found that F. hepatica infection induces the differential expression of 128 genes in the liver in the early stage of infection and 308 genes in the late stage, and most of them are up-regulated. The Ingenuity Pathway Analysis revealed significant changes in the pathways related to metabolism, biosynthesis and signaling as well as genes implicated in inducing liver-toxicity, injury and death. The present study provides us insights at the molecular level about the underlying mechanisms used by F. hepatica, leading to liver damage and its subsequent pathophysiology. The expression pattern obtained here could also be used to explain the lack of association between infection with F. hepatica and cholangiocarcinoma. However
Khawsuk, Witoon; Soonklang, Nantawan; Grams, Rudi; Vichasri-Grams, Suksiri; Wanichanon, Chaitip; Meepool, Ardool; Chaithirayanon, Kulathida; Ardseungneon, Pissanee; Viyanant, Vithoon; Upathum, Suchart Edward; Sobhon, Prasert
A monoclonal antibody (MoAb) against a recombinant glutathione S-transferase (rGST) of F. gigantica was produced in BALB/c mice. Reactivity and specificity of this monoclonal antibody was assessed by ELISA and immunoblotting. Six stable clones, namely 3A3, 3B2, 3C6, 4A6, 4B1 and 4D6 were obtained, All these MoAb reacted with rGST and native GST at a molecular weight of 28 kDa and found to be IgG1, kappa-light chain isotypes. These MoAb cross-reacted with Schistosoma mansoni and Schistosoma japonicum antigens at molecular weights of 28 and 26 kDa, respectively, but no cross-reactions were detected with antigens of Eurytrema and Paramphistomum spp. The localization of GST in metacercaria, 7-week-old juvenile and adult F. gigantica was performed by immunofluorescence technique, using MoAb as well as polyclonal antibody (PoAb) to the native protein as probes. In general, all clones of MoAb gave similar results and the pattern was quite similar to staining by PoAb. The fluorescence was intense, which implied the presence of a high concentration of GST in the parenchymal tissue in all stages of the parasite. However, the parenchymal cells were not evenly stained which implied the existence of subpopulations of this cell type with regard to GST production and storage. In addition, in adult and juvenile stages a moderate fluorescence was present in the basal layer of the tegument, while light fluorescence was observed in the caecal epithelium, cells in the ovary, testis and vitelline gland of the adult. In the metacercaria stage, in addition to parenchymal tissue, the tegument and tegumental cells were stained relatively more intense with MoAb and PoAb than in other stages.
Kueakhai, Pornanan; Meemon, Krai; Changklungmoa, Narin; Chaithirayanon, Kulathida; Riengrojpitak, Suda; Sobhon, Prasert
Fasciola gigantica saposin-like protein-2 (FgSAP-2) belongs to a family of lipid-interacting proteins that are involved in the cytolysis of target cells. In this study, we have cloned and expressed FgSAP-2 and produced the antibody against this recombinant protein. Rabbit antiserum against rFgSAP-2 reacted with a similar native protein in the whole body extracts of the 4-week-old juvenile and adult stage, as well as a protein in their excretion-secretion, but not in the tegument. In situ hybridization and immunofluorescence detection revealed the presence of SAP-2 mRNA transcripts and proteins in the cecal epithelial cells of 4-week-old juvenile and adult parasites, but not in the metacercariae and newly excysted juveniles. Moreover, SAP-2 is present only in the cecal epithelial cells lining the distal part of the digestive tract, but not in the tegumental-type epithelium lining the proximal part of the digestive tract. The rFgSAP-2 reacted with antisera from rabbits infected with F. gigantica metacercariae collected at 5 weeks, but not at 2 weeks after infection. Anti-rFgSAP-2 did not exhibit any cross-reactivity with the other parasites' antigens, including Opisthorchis viverrini, Eurytrema pancreaticum, Cotylophoron cotylophorum, Fischoederius cobboldi, Gigantocotyle explanatum, Paramphistomum cervi, Setaria labiato-papillosa, and Haemonchus placei. This finding indicated that SAP-2 is a unique protein that is expressed only in late juvenile and adult F. gigantica, and it could be considered for immunodiagnostic and as a vaccine candidate for fasciolosis.
Wongwairot, Sirima; Kueakhai, Pornanan; Changklungmoa, Narin; Jaikua, Wipaphorn; Sansri, Veerawat; Meemon, Krai; Songkoomkrong, Sineenart; Riengrojpitak, Suda; Sobhon, Prasert
Cathepsin Ls (CatLs), the major cysteine protease secreted by Fasciola spp., are important for parasite digestion and tissue invasion. Fasciola gigantica cathepsin L1H (FgCatL1H) is the isotype expressed in the early stages for migration and invasion. In the present study, a monoclonal antibody (MoAb) against recombinant F. gigantica cathepsin L1H (rFgCatL1H) was produced by hybridoma technique using spleen cells from BALB/c mice immunized with recombinant proFgCatL1H (rproFgCatL1H). This MoAb is an immunoglobulin (Ig)G1 with κ light chain isotype. The MoAb reacted specifically with rproFgCatL1H, the native FgCatL1H at a molecular weight (MW) 38 to 48 kDa in the extract of whole body (WB) of metacercariae and newly excysted juvenile (NEJ) and cross-reacted with rFgCatL1 and native FgCatLs at MW 25 to 28 kDa in WB of 2- and 4-week-old juveniles, adult, and adult excretory-secretory (ES) fractions by immunoblotting and indirect ELISA. It did not cross-react with antigens in WB fractions from other parasites, including Gigantocotyle explanatum, Paramphistomum cervi, Gastrothylax crumenifer, Eurytrema pancreaticum, Setaria labiato-papillosa, and Fischoederius cobboldi. By immunolocalization, MoAb against rFgCatL1H reacted with the native protein in the gut of metacercariae and NEJ and also cross-reacted with CatL1 in 2- and 4-week-old juveniles and adult F. gigantica. Therefore, FgCatL1H and its MoAb may be used for immunodiagnosis of both early and late fasciolosis in ruminants and humans.
Jaikua, Wipaphorn; Kueakhai, Pornanan; Chaithirayanon, Kulathida; Tanomrat, Rataya; Wongwairot, Sirima; Riengrojpitak, Suda; Sobhon, Prasert; Changklungmoa, Narin
Superoxide dismutases (SOD), antioxidant metallo-enzymes, are a part of the first line of defense in the trematode parasites which act as the chief scavengers for reactive oxygen species (ROS). A recombinant Fasciola gigantica cytosolic SOD (FgSOD) was expressed in Escherichia coli BL21 (DE3) and used for immunizing rabbits to obtain polyclonal antibodies (anti-rFgSOD). This rabbit anti-rFgSOD reacted with the native FgSOD at a molecular weight of 17.5kDa. The FgSOD protein was expressed at high level in parenchyma, caecal epithelium and egg of the parasite. The rFgSOD reacted with antisera from rabbits infected with F. gigantica metacercariae collected at 2, 5, and 7 weeks after infection, and reacted with sera of infected mice. Anti-rFgSOD exhibited cross reactivity with the other parasites' antigens, including Eurytrema pancreaticum, Cotylophoron cotylophorum, Fischoederius cobboldi, Gastrothylax crumenifer, Paramphistomum cervi, and Setaria labiato papillosa. A vaccination was performed in imprinting control region (ICR) mice by subcutaneous injection with 50μg of rFgSOD combined with Freund's adjuvant. At 2 weeks after the second boost, mice were infected with 15 metacercariae by oral route. IgG1 and IgG2a in the immune sera were determined to indicate Th2 and Th1 immune responses. It was found that the parasite burden was reduced by 45%, and both IgG1 and IgG2a levels showed correlation with the numbers of worm recoveries. Copyright © 2016 Elsevier B.V. All rights reserved.
Anuracpreeda, Panat; Songkoomkrong, Sineenart; Sethadavit, Manussabhorn; Chotwiwatthanakun, Charoonroj; Tinikul, Yotsawan; Sobhon, Prasert
A number of monoclonal antibodies (MoAbs) against a recombinant cathepsin B3 (rCatB3) of Fasciola gigantica were produced in BALB/c mice. Reactivity and specificity of these MoAbs were assessed by indirect ELISA and immunoblotting techniques. Six stable clones, namely 1C4, 1E9, 2E5, 2F9, 5B4, 5D7 were obtained. All MoAbs reacted with rCatB3 at molecular weight (MW) 37 kDa as well as the glycosylated peptide at 55-75 kDa and with the native CatB3 at MW 37 kDa in WB extracts of metacercariae (Met) and newly excysted juveniles (NEJ). It was found to be IgG(1) and λ light chain isotypes. Immunolocalization of CatB3 in metacercariae, NEJ, 4-week-old juvenile and adult F. gigantica performed by immunoperoxidase technique by using these MoAbs as probes indicated that CatB3 was present in high concentration in the caecal epithelium and caecal lumen of the Met and NEJ, but not in the 4-week-old juvenile and adult fluke. The MoAbs show no cross-reactions with antigens of other parasites including Gigantocotyl explanatum, Eurytrema pancreaticum, Paramphistomum cervi, Schistosoma spindale, S. mansoni, Haemonchus placei and Setaria labiato-papillosa. Thus, it is possible that these MoAbs could be a good candidate for immunodiagnosis of fasciolosis. Copyright © 2010 Elsevier Inc. All rights reserved.
Sultanov, A; Abdybekova, A; Abdibaeva, A; Shapiyeva, Z; Yeshmuratov, T; Torgerson, P R
Fish borne trematodes are an important cause of morbidity in Kazakhstan. The number of human cases of opisthorchiidosis (infection with parasites of the family opisthorchiidae) reached a peak of 2521 recorded cases (17 cases per 100,000) in 2002 with a gradual decline to 1225 cases (7.4 cases per 100,000) in 2011. Most human cases are found in the north and north east part of Kazakhstan in areas drained by the Irtysh River and it tributaries. A further focus is found in the north west in the Ural river basin in the European part of Kazakhstan. The most common occupations of patients with opisthorchiidosis included the manual laborers, those employed in the home or unemployed. Necropsy investigations of village dogs in an endemic region revealed 37 of 51 (72%) village dogs infected with either Opisthorchis felineus or Methorchis bilis. Likewise an investigation of 242 cats consisting of strays, village, suburban and city cats revealed 79 (33%) animals infected with O. felineus. Higher prevalences were seen in strays and village cats compared to suburban cats. No urban cats, which lived in apartments, were found to be infected. Other important zoonoses included Echinococcus granulosus, detected in 2 of the 51 necropsied village dogs and E. multilocularis was found in 2 out of 124 necropsied stray cats. Investigations of locally caught fish revealed 10 of 107 (9%) roach (Rutilus rutilus), 49 of 68(72%) ide (Leuciscus idus) and 2 of 79 (2.5%) bream (Abramis brama) infested with trematode metacercariae. No metacercariae were found in 609 crucian carp (Carassius carassius), 35 tench (Tinca tinca), 79 carp (Cyprinus carpio), 46 perch (Perca fluviatilis) or 20 zander (Sander lucioperca). Copyright © 2014 Elsevier B.V. All rights reserved.
Butcher, A R; Brealey, J K; Grove, D I; Dymock, R B
Brachylaima cribbi is a recently described species of terrestrial trematode that infects mammals and birds with helicid land snails as its first and second intermediate hosts. The adult worm is 2.5-6.0 mm long by 0.5-0.8 mm wide being a long slender cylindrical worm with oral and ventral suckers in the anterior quarter and genital pore in the posterior quarter. Scanning electron microscopy shows that there is a dense covering of tegumental spines at the anterior end which diminishes towards the posterior extremities of the worm. Development of spines was observed in juvenile and mature adult worms. In young worms 1-3 weeks post infection (wpi) spines appear as buds with a serrated edge each having 1-4 spikes per spine. As the worm ages the spines broaden and by 5 wpi the number of spikes per spine increases to an average of 8.1. The serial development of oral sucker papillae in the cercaria, metacercaria and adult worm was observed with the finding of an elongated papilla with a bifurcated tip on the cercaria becoming a shorter and thicker elongated papilla with a large central stoma on the metacercaria. In the adult worm, this papilla becomes dome-shaped with a small central stoma. For some of these papillae a cilium could be seen extended from the central stoma. Other life-cycle stages illustrated were the hatched egg with an extruded egg membrane minus an operculum and a portion of the branched sporocyst dissected from the digestive gland of the land snail Theba pisana showing a terminal birth pore. Scanning electron microscopy morphological features of the adult worm observed for the first time in a Brachylaima were the unarmed cirrus extended from the genital pore with released sperm present and the Laurer's canal opening visible in tegumental folds on the dorsal surface approximately 300 microm posterior to the genital pore.
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. Copyright © 2013 Elsevier B.V. All rights reserved.
Full Text Available El objetivo del trabajo fue realizar un estudio de foco relacionado con un caso de fasciolosis humana ocurrido en abril de 2002 en Loncopué, Neuquén, Argentina. La confirmación diagnóstica se efectuó en mayo de 2002 por el test de ELISA. En noviembre de 2002 se realizó un muestreo en el área rural donde se ubicaba la vivienda de la paciente, ya restablecida, y se le tomó una nueva muestra de sangre. El suero de la paciente continuó reactivo para antígenos de Fasciola hepatica. Se muestrearon plantas de berro para detectar metacercarias. Se recolectaron caracoles en cuatro canales de riego conectados a un canal principal. Los caracoles fueron trasladados vivos para su identificación, medición y examen de infección. Se recolectaron 35 muestras fecales de ganado de cría. No se observaron metacercarias en las hojas de berro examinadas (n=222. Se recolectaron 130 caracoles identificados como Lymnaea viatrix y 2 de 101 ejemplares (2% estaban infectados con larvas de F. hepatica. Las prevalencias en el ganado adulto fueron: 100% (10/10 para caprinos, 82% (9/11 para ovinos y 86% (6/7 para bovinos. El número de huevos eliminados por las cabras (mediana = 20.7; Q1=6.2; Q3=34.5 y ovejas (4, 18, 13 infectadas, resultó mayor que el eliminado por vacas (0.3; 0.3; 1.7 (pAn epidemiological focal study was performed in Loncopué, Neuquén, Argentina, in November 2002 to detect the origin of the infection in a human case of fascioliasis confirmed by an indirect-ELISA test, six months before the study. Thirty five individual fecal samples were taken from domestic livestock, and watercress plants and snails were collected from the irrigation ditches connected to a main canal in the surroundings of the patient’s house. A new blood sample was taken from the already recovered patient. The patient was still seropositive to Fasciola hepatica antigens. No metacercariae were found in the 222 watercress leaves checked. All the snails collected (n=130 were
Full Text Available During the period between January 1999 and December 2000, the distribution and seasonal patterns of Fasciola gigantica infections in cattle in the highveld and lowveld communal grazing areas of Zimbabwe were determined through monthly coprological examination. Cattle faecal samples were collected from 12 and nine dipping sites in the highveld and lowveld communal grazing areas respectively. Patterns of distribution and seasonal fluctuations of the intermediate host-snail populations and the climatic factors influencing the distribution were also determined by sampling at monthly intervals for a period of 24 months (November 1998 to October 2000 in six dams and six streams in the highveld and in nine dams in the lowveld communal grazing areas. Each site was sampled for relative snail density and the vegetation cover and type, physical and chemical properties of water, and mean monthly rainfall and temperature were recorded. Aquatic vegetation and grass samples 0-1 m from the edges of the snail habitats were collected monthly to determine the presence or absence of F. gigantica metacercariae. Snails collected at the same time were individually checked for the emergence of larval stages of F. gigantica. A total of 16 264 (calves 5 418; weaners 5 461 and adults 5 385 faecal samples were collected during the entire period of the study and 2 500 (15.4 % of the samples were positive for F. gigantica eggs. Significantly higher prevalences were found in the highveld compared to the lowveld (P < 0.001, for adult cattle than calves ( P < 0.01 and in the wet season over the dry season (P < 0.01. Faecal egg output peaked from August / September to March / April for both years of the study. Lymnaea natalensis, the snail intermediate host of F. gigantica was recorded from the study sites with the highveld having a significantly higher abundance of the snail species than the lowveld (P < 0.01. The snail population was low between December and March and started
Full Text Available During the period between January 1999 and December 2000, the distribution and seasonal patterns of amphistome infections in cattle in the highveld and lowveld communal grazing areas of Zimbabwe were determined through monthly coprological examination. Cattle faecal samples were collected from 12 and nine dipping sites in the highveld and lowveld communal grazing areas, respectively. Patterns of distribution and seasonal fluctuations of intermediate host-snail populations and the climatic factors influencing the distribution were also determined by sampling at monthly intervals for a period of 24 months (November 1998 to October 2000 in six dams and six streams in the highveld and in nine dams in the lowveld communal grazing areas. Each site was sampled for relative snail density and the vegetation cover and type, physical and chemical properties of water, and mean monthly rainfall and temperature were recorded. Aquatic vegetation and grass samples 0-1 m from the edges of the snail habitats were collected monthly to determine the presence or absence of amphistome metacercariae. Snails collected at the same time were individually checked for the emergence of larval stages of amphistomes. A total of 16 264 (calves 5 418, weaners 5 461 and adults 5 385 faecal samples were collected during the entire period of the study and 4 790 (29.5 % of the samples were positive for amphistome eggs. For both regions the number of animals positive for amphistome eggs differed significantly between the 2 years, with the second year having a significantly higher prevalence (P < 0.01 than the first year. Significantly higher prevalences were found in the highveld compared to the lowveld (P < 0.001, for adult cattle than calves (P < 0.01, and in the wet over the dry season (P < 0.01. Faecal egg output peaked from October to March in both years of the study. Bulinus tropicus, Bulinus forskalii and Biomphalaria pfeifferi were recorded from the study sites. The main
Full Text Available Fasciola hepatica infection still remains one of the helminthic neglected tropical diseases (NTDs. It has a huge worldwide distribution, affecting mainly cattle and, sometimes, human beings. In addition to data reported about the immunological response induced by helminthic infections and that induced by Fasciola hepatica, little is known about the gene expression profile in its organ target, the liver, which is where adult worms are established and live for long periods of time, causing its characteristic pathology. In the present work, we study both the early and late gene expression profiles in the livers of mice infected with F. hepatica metacercariae using a microarray-based methodology.A total of 9 female-6-week-old BALB/c mice (Charles River Laboratories, Barcelona, Spain weighing 20 to 35 g were used for the experiments. Two groups of BALB/c mice were orally infected with seven F. hepatica metacercariae, and the other group remained untreated and served as a control. Mice were humanely euthanized and necropsied for liver recovery, histological assessment of hepatic damage, RNA isolation, microarray design and gene expression analysis on the day of infection (t0, seven days post-infection (t7 and twenty-one days post-infection (t21.We found that F. hepatica infection induces the differential expression of 128 genes in the liver in the early stage of infection and 308 genes in the late stage, and most of them are up-regulated. The Ingenuity Pathway Analysis revealed significant changes in the pathways related to metabolism, biosynthesis and signaling as well as genes implicated in inducing liver-toxicity, injury and death.The present study provides us insights at the molecular level about the underlying mechanisms used by F. hepatica, leading to liver damage and its subsequent pathophysiology. The expression pattern obtained here could also be used to explain the lack of association between infection with F. hepatica and cholangiocarcinoma
Full Text Available Abstract Background Paragonimiasis have previously been reported in two zones of the Southwest Province of Cameroon including the Kupe mountain and Mundani foci. The aim of this study was to investigate the presence and epidemiology of paragonimiasis in the peri-urban zone of Kumba, Meme Division, located about 50 km away from the Kupe mountain focus. Methods Pupils of several government primary schools in 5 villages around Kumba underwent both parasitologic and clinical investigations in search of signs and symptoms of paragonimiasis. Mycobacterium tuberculosis was also searched for in the differential diagnosis.Freshwater crabs from neighbouring streams in the five villages were dissected in search of paragonimus metacercariae. Results Out of a total of 1482 pupils examined in all five villages, 309 individuals (147 males and 162 females were recruited for this study based on the presence of one or more signs or symptoms of paragonimiasis. Eggs of Paragonimus africanus were found in stools and/or sputum of pupils from all five villages, giving an overall paragonimus prevalence of 2.56%. There was no significant difference in the disease prevalence between the villages (X2 = 8.36, P = 0.08. The prevalence of Paragonimus africanus eggs amongst pupils with symptoms of paragonimiasis was 12.3% (38 of 309. Males were infected more than females (17.0% versus 8.0%, but the difference was not significant (X2 = 5.76, P = 0.16. All the 38 paragonimus egg positive subjects presented with cough, 23 (60.53% complained of chest pain while 16 (42.11% had haemoptysis. Stool examinations also detected some intestinal parasites including Ascaris lumbricoides (29.45%, Trichuris trichiura (6.47%, Necator americanus (2.27%, Strongyloides stercoralis (1.62%, Enterobius vermicularis (0.65%, and Entamoeba histolytica (4.53%. No case of M. tuberculosis was noted. Out of a total of 85 dissected crabs (Sudanonautes africanus, 6.02 % were infected with paragonimus
Tang, Zeli; Shang, Mei; Chen, Tingjin; Ren, Pengli; Sun, Hengchang; Qu, Hongling; Lin, Zhipeng; Zhou, Lina; Yu, Jinyun; Jiang, Hongye; Zhou, Xinyi; Li, Xuerong; Huang, Yan; Xu, Jin; Yu, Xinbing
Clonorchiasis, a food-borne zoonosis, is caused by Clonorchis sinensis. The intestinal tract and bile ducts are crucial places for C. sinensis metacercariae to develop into adult worms. The endospore of Bacillus subtilis is an ideal oral immunization vehicle for delivery of heterologous antigens to intestine. Cysteine protease of C. sinensis (CsCP) is an endogenous key component in the excystment of metacercariae and other physiological or pathological processes. We constructed a fusion gene of CotC (a coat protein)-CsCP and obtained B. subtilis spores with recombinant plasmid of pEB03-CotC-CsCP (B.s-CotC-CsCP). CotC-CsCP expressed on spores' surface was detected by Western blotting and immunofluorescence. Immunological characteristics of recombinant spore coat protein were evaluated in a mouse model. The levels of CsCP-specific antibodies were detected by ELISA. Effects of recombinant spores on mouse intestine were evaluated by histological staining. The activities of biochemical enzymes in serum were assayed by microplate. Liver sections of infected mice were evaluated by Ishak score after Masson's trichrome. The B.s-CotC-CsCP spores displayed CsCP on their coat. Specific IgG and isotypes were significantly induced by coat proteins of B.s-CotC-CsCP spores after subcutaneous immunization. IgA levels in intestinal mucus and bile of B.s-CotC-CsCP orally treated mice significantly increased. Additionally, more IgA-secreting cells were observed in enteraden and lamina propria regions of the mouse jejunum, and an increased amount of acidic mucins in intestines were also observed. There were no significant differences in enzyme levels of serum among groups. No inflammatory injury was observed in the intestinal tissues of each group. The degree of liver fibrosis was significantly reduced after oral immunization with B.s-CotC-CsCP spores. Bacillus subtilis spores maintained the original excellent immunogenicity of CsCP expressed on their surface. Both local and systemic
Anuracpreeda, Panat; Srirakam, Thippawan; Pandonlan, Sudarat; Changklungmoa, Narin; Chotwiwatthanakun, Charoonroj; Tinikul, Yotsawan; Poljaroen, Jaruwan; Meemon, Krai; Sobhon, Prasert
Monoclonal antibodies (MoAbs) against a recombinant cathepsin L1 of Fasciola gigantica (rFgCatL1) were produced in vitro by fusion of BALB/c mice spleen cells immunized with rFgCatL1 and mouse myeloma cells. Reactivity and specificity of these MoAbs were evaluated by indirect ELISA and immunoblotting techniques. Seven MoAb clones were selected from the stable hybridoma clones, namely 1E10, 1F5, 3D11, 4B10, 4D3, 4E3 and 5E7. Clones 1E10, 1F5 and 3D11 were IgM, whereas clones 4B10, 4D3, 4E3 and 5E7 were IgG1. All MoAbs had kappa light chain isotypes. All MoAbs reacted with rCatL1 at molecular weight (MW) 30kDa and with the native CatL1 at MW 27kDa in whole body (WB) extracts of metacercariae (Met), newly excysted juveniles (NEJ), 1, 3, 5-week-old juveniles (Ju), adult WB and adult excretory-secretory (ES) fractions, but not with adult tegumental antigens (TA). All of these MoAbs showed no cross-reactions with antigens of other parasites commonly found in ruminants and human, including Paramphistomum cervi, Eurytrema pancreaticum, Gigantocotyle explanatum, Schistosoma spindale, Schistosoma mansoni, Moniezia benedeni, Avitellina centripunctata, Trichuris sp., Haemonchus placei and Setaria labiato-papillosa. Localization of CatL1 in each developmental stages of F. gigantica by immunoperoxidase technique, using these MoAbs as probes, indicated that CatL1 was present at high concentration in the caecal epithelium and caecal lumen of metacercariae, NEJ, 1, 3, 5-week-old juveniles and adult fluke. This finding indicated that CatL1 is a copiously expressed parasite protein that is released into the ES, thus CatL1 and its MoAb could be a good candidate for immunodiagnosis of fasciolosis in ruminant and human. Copyright © 2014 Elsevier B.V. All rights reserved.
Ondračková, Markéta; Hudcová, Iveta; Dávidová, Martina; Adámek, Zdeněk; Kašný, Martin; Jurajda, Pavel
Introduced species can modify local host-parasite dynamics by amplifying parasite infection which can 'spill-back' to the native fauna, whether they are competent hosts for local parasites, or by acting as parasite sinks with 'dilution' of infection decreasing the parasite burden of native hosts. Recently infection by the trematode Bucephalus polymorphus has increased in several European rivers, being attributed to the introduction of intermediate host species from the Ponto-Caspian region. Using a combination of field and experimental data, we evaluated the competence of non-native and native fish as intermediate hosts for B. polymorphus and its role for parasite development in a definitive host. The density of 0+ juvenile fish (the second intermediate hosts for B. polymorphus) was measured in the River Morava, Czech Republic and fish were screened for natural metacercariae infection. The stomach contents of predatory fish that are definitive hosts of B. polymorphus were examined to assess the importance of non-native gobies for parasite transmission. In semi-natural conditions, parasite establishment, initial survival, and maturity rates in experimentally infected definitive hosts pikeperch Sander lucioperca were measured in flukes recovered from native white bream Abramis bjoerkna and non-native tubenose goby Proterorhinus semilunaris and round goby Neogobius melanostomus. Adult fluke size and egg production was also measured to evaluate the potential effect of intermediate host species on parasite fitness. We detected high natural infection parameters of B. polymorphus in native cyprinids and non-native gobies compared to data from the period prior to goby establishment. Both fish groups are consumed by predatory fish and represent a major component of the littoral fish community. Parasite establishment and adult size in definitive hosts was equivalent among the second intermediate host species, despite a lower size of metacercariae recovered from round gobies
Full Text Available Clonorchiasis, which is induced by the infection of Clonorchis sinensis (C. sinensis, is highly associated with cholangiocarcinoma. Because the available examination, treatment and interrupting transmission provide limited opportunities to prevent infection, it is urgent to develop integrated strategies to prevent and control clonorchiasis. Glycolytic enzymes are crucial molecules for trematode survival and have been targeted for drug development. Hexokinase of C. sinensis (CsHK, the first key regulatory enzyme of the glycolytic pathway, was characterized in this study. The calculated molecular mass (Mr of CsHK was 50.0 kDa. The obtained recombinant CsHK (rCsHK was a homotrimer with an Mr of approximately 164 kDa, as determined using native PAGE and gel filtration. The highest activity was obtained with 50 mM glycine-NaOH at pH 10 and 100 mM Tris-HCl at pH 8.5 and 10. The kinetics of rCsHK has a moderate thermal stability. Compared to that of the corresponding negative control, the enzymatic activity was significantly inhibited by praziquantel (PZQ and anti-rCsHK serum. rCsHK was homotropically and allosterically activated by its substrates, including glucose, mannose, fructose, and ATP. ADP exhibited mixed allosteric effect on rCsHK with respect to ATP, while inorganic pyrophosphate (PPi displayed net allosteric activation with various allosteric systems. Fructose behaved as a dose-dependent V activator with the substrate glucose. Glucose-6-phosphate (G6P displayed net allosteric inhibition on rCsHK with respect to ATP or glucose with various allosteric systems in a dose-independent manner. There were differences in both mRNA and protein levels of CsHK among the life stages of adult worm, metacercaria, excysted metacercaria and egg of C. sinensis, suggesting different energy requirements during different development stages. Our study furthers the understanding of the biological functions of CsHK and supports the need to screen for small
Full Text Available Los crustáceos tienen un rol importante en el ciclo de vida de los parásitos, ya que actúan como hospedadores intermediarios. No obstante, en Chile y a nivel mundial existen pocos estudios parasitológicos sobre crustáceos que habitan el intermareal rocoso. En este estudio se registró y comparó la carga parasitaria de crustáceos decápodos, y se relacionó con la abundancia de sus hospedadores definitivos (peces y aves. Entre julio y septiembre de 2013 se recolectaron 409 crustáceos, correspondientes a 16 especies, desde el intermareal rocoso de dos localidades de Chile central (33°S, Las Cruces y Montemar. El 65,5% de la muestra estaba parasitada, recolectándose 2.410 metacercarias y 18 nemátodos. Algunas metacercarias correspondían a la familia Opecoelidae, mientras que otras a Microphallidae; los nematodos eran de la familia Cystidicolidae. La mayor prevalencia y abundancia de Microphallidae se registró en Petrolisthes tuberculosus (42,3%; 4,8 ± 11,08 parásitos/crustáceo y de Opecoelidae en P. violaceus (96,9%; 13,59 ± 17,50 parásitos/crustáceo, mientras que Cystidicolidae fue poco prevalente y abundante en ambas localidades. Las infecciones parasíticas fueron afectadas por la localidad de muestreo, especie y tamaño del hospedador. No se encontró asociación entre la abundancia de los hospedadores definitivos y las cargas parasitarias que presentaban los crustáceos. La nula relación entre las cargas parasitarias y la abundancia de hospedadores definitivos puede ocurrir si estos últimos tienen un amplio espectro trófico o viajan largas distancias, de este modo, la transmisión de los parásitos no sería afectada directamente por la abundancia de los hospedadores definitivos.
The life cycle of Bucephalus margaritae Ozaki & Ishibashi, 1934 (Digenea, Bucephalidae from the coast of Santa Catarina State, Brazil = O ciclo de vida de Bucephalus margaritae Ozaki e Ishibashi, 1934 (Digenea, Bucephalidae da costa do Estado de Santa Catarina, Brasil
Natalia da Costa Marchiori
Full Text Available The orange disease is considered the main parasitosis in Perna perna mussel. It is caused by a complex life cycle bucephalid, involving three hosts, among them mollusks and fishes. With the aim of contributing to the knowledge of orange disease in mussel culture, the parasite life cycle was investigated. Experimental studies and monthly samples in the study area allowed the identification and characterization of the Bucephalus margaritae life cycle. Larvae and adults of B. margaritae were fixed in 5% formaldehyde, stained with Gomori’s trichrome, clarified in creosote and mounted in Canada balsam. The cercariae are found in the first intermediate host P. perna inside the sporocysts, which have the form of orange and ramified filaments. The metacercariae encysts in the gills and gill cavity of the second intermediate host, the blenniid Hypleurochilus fissicornis. The definitive host Menticirrhus americanus is infected ingesting blenniids parasitized with metacercariae. The high parasitological indexes of B. margaritae suggests that M. americanus and H. fissicornis act as the main definitive and intermediate hosts, respectively, in the trematode life cycle. The blenniid H. fissicornis is a new intermediate host to the species.A bucefalose é considerada a principal parasitose do mexilhão Perna perna. É causada por um Bucephalidae, de ciclo de vida complexo, envolvendo três hospedeiros, entre eles, moluscos e peixes. Com o objetivo de se contribuir para o conhecimento da bucefalose em mexilhões de cultivo, foi investigado o ciclo de vida deste parasito. Estudos experimentais e coletas mensais na área de estudo permitiram caracterizar o ciclo de vida de Bucephalus margaritae. Larvas e adultos do parasito foram fixados em formol 5%, corados com tricrômico de Gômori, clarificados em creosoto e montados em lâminas permanentes com bálsamo do Canadá. As cercárias ocorrem no primeiro hospedeiro intermediário P. perna, no interior dos
Melanoides tuberculata (Mollusca: Thiaridae as an intermediate host of Centrocestus formosanus (Trematoda: Heterophyidae in Brazil Melanoides tuberculata (Mollusca: Thiaridae como hospedeiro intermediário de Centrocestus formosanus (Trematoda: Heterophyidae no Brasil
Hudson Alves Pinto
Full Text Available Pleurolophocercous cercariae emerged from naturally infected Melanoides tuberculata from Minas Gerais State, Brazil, were used to perform experimental infection of laboratory-reared Poecilia reticulata. Mature metacercariae were obtained from the gills of fishes and force-fed to Mus musculus. The adult parasites which recovered from small intestines of mice were identified as Centrocestus formosanus. This is the first report of M. tuberculata as intermediate host of this heterophyid in Brazil.Cercárias do tipo pleurolofocerca emergidas de Melanoides tuberculata naturalmente infectados coletados na represa da Pampulha, Belo Horizonte, Minas Gerais, Brasil, foram utilizadas para a infecção experimental de Poecilia reticulata criados em laboratório. Metacercárias maduras foram obtidas nas brânquias dos peixes e administradas por via oral a Mus musculus. Parasitos adultos recuperados no intestino delgado dos camundongos foram identificados como Centrocestus formosanus. Este é o primeiro relato de M. tuberculata como hospedeiro intermediário deste parasito no Brasil.
Dusak, Abdurrahim; Onur, Mehmet R; Cicek, Mutalip; Firat, Ugur; Ren, Tianbo; Dogra, Vikram S
Fascioliasis refers to a zoonosis caused by Fasciola hepatica, a trematode infecting herbivores, but also occurs in humans who ingest the metacercaria found in fresh water plants. Infection in humans is common in developing countries and is also not uncommon in Europe. Diagnosis of this infection is difficult, as the history and symptoms are nonspecific and stool analysis for eggs is negative until the disease is in an advanced state by when the parasite has reached the biliary system. The clinical course consists of two phases; first a hepatic parenchymal phase in which immature larvae invade the liver parenchyma, followed by a ductal phase characterized by the excretion of larvae into the bile ducts. Parenchymal Phase: Ultrasonography (US) findings are nonspecific in this early phase. Computerized tomography (CT) may demonstrate subcapsular low attenuation regions in the liver. Magnetic Resonance imaging (MRI) can also be utilized to establish liver parenchymal involvement, and is better than CT in characterizing hemorrhagic lesions, as well as identifying more lesions relative to CT. Ductal Phase: US examination is most useful at this stage, with its ability to demonstrate the live movement of the worms within the dilated ducts. A CT demonstrates dilated central biliary ducts with periportal tracking, whereas, mild ductal dilatation is poorly appreciated under MRI. Therefore, familiarity with the multimodality imaging features of fascioliasis, in combination with an available confirmatory enzyme-linked immunoassay, would be most helpful for early diagnosis.
Edith, R; Godara, R; Sharma, R L; Thilagar, M B
Sixteen male, yearling Murrah buffaloes were randomly assigned to four groups of four buffaloes each. Each animal in Gr-I and II were immunized, respectively, with 4.8 mg of excretory secretory antigen and 1,300 microg Infection Specific Antigen, in three divided doses. Subsequently, each animal in Gr-I, II, and III was per os infected with 800 viable Fasciola gigantica metacercariae (bubaline origin) on week-6 after the start of the experiment, while animals in Gr-IV served as healthy controls. The aspartate aminotransferase (AST) and alanine aminotransferase (ALT) concentrations progressively increased during the prepatency and respectively attained the highest levels during week-6 and 8 post-infection (PI). The alkaline phosphatase (AP) exhibited elevated trends from eighth week PI onward and continued to be higher until the end (p hepatic lesion score in the respective groups. The increased concentrations of the enzymes revealed two clearly demarcated stages: (a) remarkably elevated AST (40.8%) and ALT (140.0%) levels during the prepatency, signifying traumatic lesions inflicted by the F. gigantica adolescercariae and (b) the significant increase in AP (107.9%), suggestive of bile duct hyperplasia, cholangitis, periportal fibrosis, and biliary obstruction etc. from sixth week PI onward. None of healthy controls developed clinical signs and had normal hematological and serum enzyme profiles. Diagnostic significance of these marker enzymes in the disease forecasting and in time application of control strategies to combat tropical fasciolosis in buffaloes in the endemic areas has been discussed.
Bottari, Nathieli B; Mendes, Ricardo E; Lucca, Neuber J; Schwertz, Claiton I; Henker, Luan C; Olsson, Débora C; Piva, Manoela M; Sangoi, Manuela; Campos, Luízi P; Moresco, Rafael N; Jaques, Jeandre A; Da silva, Aleksandro S
The aim of this study was to analyze the antioxidant status and oxidative profile in serum and liver of rats experimentally infected with Fasciola hepatica and its relationship with pathological findings. Twenty-four rats were divided into two groups: group A consisted of 12 healthy rats and group B of 12 rats infected orally with 20 metacercaria of F. hepatica. At days 20 and 150 post-infection (PI), blood and liver samples of six animals from each group were collected. The protein oxidation (AOPP technique: advanced oxidation protein products) and antioxidants (FRAP technique: ferric reducing antioxidant power) levels were measured in serum and liver. Furthermore, nitrite/nitrate (NOx) levels and lipid peroxidation (TBARS technique: thiobarbituric acid reactive substances) were measured in liver. AOPP and FRAP levels were increased (P hepatitis, composed of an abundant inflammatory infiltrate in portal spaces on infected animals, as well as bile duct hyperplasia. The results found seem to be related to the host free radical production demonstrated in serum samples and liver due to the parasite infection. Copyright © 2015 Elsevier Inc. All rights reserved.
Dracz, Ruth Massote; Ribeiro, Vinicius Marques Antunes; Pereira, Cintia Aparecida de Jesus; Lima, Walter Dos Santos
Fasciola hepatica is a parasite that affects the hepatic ducts of several species of domestic and wild vertebrates, causing huge economic losses to livestock rearing worldwide. Reports on occurrences of F. hepatica in capybaras are an important epidemiological aspect of this disease, since these rodents can be a source of contamination for other animals and humans. In the present study, conducted in a rural area of the municipality of Confins, Minas Gerais, fresh feces from capybaras were collected from the ground near a lagoon at the edge of the Ribeirão da Mata river. These were examined using the technique of four metal sieves. F. hepatica eggs were recovered. This trematode species was confirmed by observing morphological characteristics and measuring the eggs recovered from the capybara feces, and through experimental infection of Lymnaea columella (Say, 1817) by miracidia from these eggs and subsequent infection of C57/BL06 mice with metacercariae originating from these infected mollusks. The data suggest the occurrence of natural cycle of F. hepatica in this region and provide a warning that expansion of the geographical distribution of this parasite by means of this rodent is possible. It is therefore important to adopting measures for epidemiological control of this helminthiasis.
Ruth Massote Dracz
Full Text Available Abstract Fasciola hepatica is a parasite that affects the hepatic ducts of several species of domestic and wild vertebrates, causing huge economic losses to livestock rearing worldwide. Reports on occurrences of F. hepatica in capybaras are an important epidemiological aspect of this disease, since these rodents can be a source of contamination for other animals and humans. In the present study, conducted in a rural area of the municipality of Confins, Minas Gerais, fresh feces from capybaras were collected from the ground near a lagoon at the edge of the Ribeirão da Mata river. These were examined using the technique of four metal sieves. F. hepatica eggs were recovered. This trematode species was confirmed by observing morphological characteristics and measuring the eggs recovered from the capybara feces, and through experimental infection of Lymnaea columella (Say, 1817 by miracidia from these eggs and subsequent infection of C57/BL06 mice with metacercariae originating from these infected mollusks. The data suggest the occurrence of natural cycle of F. hepatica in this region and provide a warning that expansion of the geographical distribution of this parasite by means of this rodent is possible. It is therefore important to adopting measures for epidemiological control of this helminthiasis.
Full Text Available Fascioliasis refers to a zoonosis caused by Fasciola hepatica, a trematode infecting herbivores, but also occurs in humans who ingest the metacercaria found in fresh water plants. Infection in humans is common in developing countries and is also not uncommon in Europe. Diagnosis of this infection is difficult, as the history and symptoms are nonspecific and stool analysis for eggs is negative until the disease is in an advanced state by when the parasite has reached the biliary system. The clinical course consists of two phases; first a hepatic parenchymal phase in which immature larvae invade the liver parenchyma, followed by a ductal phase characterized by the excretion of larvae into the bile ducts. Parenchymal Phase: Ultrasonography (US findings are nonspecific in this early phase. Computerized tomography (CT may demonstrate subcapsular low attenuation regions in the liver. Magnetic Resonance imaging (MRI can also be utilized to establish liver parenchymal involvement, and is better than CT in characterizing hemorrhagic lesions, as well as identifying more lesions relative to CT. Ductal Phase: US examination is most useful at this stage, with its ability to demonstrate the live movement of the worms within the dilated ducts. A CT demonstrates dilated central biliary ducts with periportal tracking, whereas, mild ductal dilatation is poorly appreciated under MRI. Therefore, familiarity with the multimodality imaging features of fascioliasis, in combination with an available confirmatory enzyme-linked immunoassay, would be most helpful for early diagnosis.
Abdel-Rahman, Eman H; Abdel-Megeed, Kadria N
Two fractions were isolated from coproantigen by ion-exchange chromatography in which DEAE cellulose was utilized. Both fractions and crude antigen were characterized by SDS-polyacrylamide gel electrophoresis which revealed 13 bands of molecular weight ranged from 205-31 in crude coproantigen. While fraction I resolved into six bands of molecular weight 198, 178, 148, 111, 101 & 45. Fraction II showed seven bands of 191 KDa, 178KDa, 166KDa, 118KDa, 98.5KDa, 72KDa & 32KDa. Fraction 1I was higher immunoreactivity than fraction by ELISA. Three immunoreactive bands of 191KDa, 118KDa & 98.5KDa were identified in fraction II using immunoblot assay. Five bands of 178KDa, 148KDa, 111KDa, 101KDa & 45KDa were detected in fraction I. Immunization of rabbits twice with fraction II in Freund's adjuvant with two weeks interval followed by challenge with F. gigantica metacercariae resulted in 66.6% protection from infection. The protection was assessed by detect-ion of hepatic damage, worm recoveries and antibody response. High level of IgG response in vaccinated rabbits than control infected ones occurred and being responsible for the recorded protection.
Calléja, C; Bigot, K; Eeckhoutte, C; Sibille, P; Boulard, C; Galtier, P
The activity of drug-metabolising enzymes was compared in liver and kidneys of adult sheep given single or two-fold fluke infection. Fascioliasis was induced by oral administration of 200 metacercariae of Fasciola hepatica to female sheep either 10 or 20 weeks (mono-infections) or 10 and 20 weeks (bi-infection) before killing. The parasitic pathology was ascertained at autopsy and by clinical observation of animals. In the liver of both mono- and bi-infected animals, significant decreases (P<0.05) (17-44%) were observed in the microsomal content of cytochrome P450 and in the two measured P450-dependent monooxygenase activities, benzphetamine and ethylmorphine N-demethylations. Moreover, Western blot analysis of microsomes demonstrated a decrease in the expression of cytochrome P4503A subfamily correlative with that of its presumed corresponding activity ethylmorphine N-demethylase. By contrast, the conjugation of chloro-dinitrobenzene to glutathione remained unchanged in liver cytosolic fractions prepared from all these animals. In kidneys, a significant decrease (P<0.05) (30%) in microsomal cytochrome P450 level of 10-week mono-infected sheep was observed whereas there was no change in the other groups of animals. The inflammatory origin and the consequences in terms of pathology and animal productivity of the fascioliasis-induced decreases in tissue-oxidative drug metabolism are discussed, particularly in the case of adult sheep suffering repetitive infections.
Full Text Available A review of national and international publications on paragonimiasis in Ecuador, epidemiological records from the Ministry of Public Health and unpublished research data was conducted to summarise the current status of the parasite/disease. The purpose of the review is to educate physicians, policy-makers and health providers on the status of the disease and to stimulate scientific investigators to conduct further research. Paragonimiasis was first diagnosed in Ecuador 94 years ago and it is endemic to both tropical and subtropical regions in 19 of 24 provinces in the Pacific Coast and Amazon regions. Paragonimus mexicanus is the only known species in the country, with the mollusc Aroapyrgus colombiensis and the crabs Moreirocarcinus emarginatus, Hypolobocera chilensis and Hypolobocera aequatorialis being the primary and secondary intermediate hosts, respectively. Recent studies found P. mexicanus metacercariae in Trichodactylus faxoni crabs of the northern Amazon. Chronic pulmonary paragonimiasis is commonly misdiagnosed and treated as tuberculosis and although studies have demonstrated the efficacy of praziquantel and triclabendazole for the treatment of human infections, neither drug is available in Ecuador. Official data recorded from 1978-2007 indicate an annual incidence of 85.5 cases throughout the 19 provinces, with an estimated 17.2% of the population at risk of infection. There are no current data on the incidence/prevalence of infection, nor is there a national control programme.
Ivan Moura Lapera
Full Text Available Abstract This study focused on the characterization and analysis of communities and infra-communities of metazoan parasites of Plagioscion squamosissimus caught in Promissão Reservoir in the Tietê River in Borborema (21°39′58”S, 49°8′49”W, state of São Paulo, Brazil. Fifty adult specimens caught by professional fishermen in March 2015 were necropsied. The fish presented an average standard length of 25.2 ± 2.2 cm and average weight of 328.82 ± 89.03 g. A total of 5,227 specimens of metazoan parasites were collected: 2,880 (55.1% adult Diplectanum piscinarius (Monogenoidea: Diplectanidae and 2,347 (44.9% Austrodiplostomum compactum metacercariae (Digenea, Diplostomidae, both with 100% prevalence and mean abundance of 57.6 and 46.9, respectively. Parasite diversity was low (species richness = 2, with a Simpson index (D equal to 0.505, and low values of Brillouin (HB = 0.687 and Margalef diversity (I = 0.117 indices. Berger-Parker’s index of dominance (p = 0.551 indicated a slight dominance of the monogenean parasite D. piscinarius. There was a positive correlation, assessed by Pearson coefficient between parasite abundance of D. piscinarius and standard length (r = 0.43 and weight (r = 0.51 of hosts.
Devi, K Rekha; Narain, Kanwar; Agatsuma, Takeshi; Blair, David; Nagataki, Mitsuru; Wickramasinghe, S; Yatawara, L; Mahanta, Jagadish
Evidence for the presence of lung flukes of the Paragonimus westermani in India remains scant. In particular, evidence based on morphology of adult worms is lacking. Metacercariae of the genus Paragonimus, recovered from crabs in two regions of northeastern India, were raised to adulthood in laboratory rats. Morphologically, these worms appear to be P. westermani. DNA sequences from the second internal transcribed spacer (ITS2) and a portion of the ribosomal large subunit gene (28S) of the nuclear ribosomal RNA gene repeat, as well as fragments of the mitochondrial cytochrome c oxidase subunit 1 (cox1) and NADH dehydrogenase subunit 1 (nad1) genes, all supported this identification. Molecular phylogenetic methods were used for studying the relatedness of these Indian flukes with counterparts from southeast and far-east Asia. Molecular data showed that Indian representatives of the P. westermani complex represent a distinct lineage. It is unclear whether the Indian form can cause disease in humans as some members of the complex do elsewhere. Copyright 2010 Elsevier B.V. All rights reserved.
Kong, Yoon; Yun, Doo-Hee; Kang, Shin-Yong; Kim, Lee-Soo; Chung, Young-Bae; Yang, Hyun-Jong
Antibody responses in serum and cerebrospinal fluid (CSF) samples from patients with active and chronic paragonimiasis and in sera from patients on whom follow-up studies were done after praziquantel treatment were analyzed using antigens of Paragonimus westermani prepared from eggs, metacercariae, juveniles of 4- and 7-week old, adult worms and recombinant protein of 28 kDa cruzipain-like cysteine protease (rPw28CCP). The patient sera/CSFs of active and chronic paragonimiasis revealed strong antibody reactions against the crude extracts of 4- and 7-week old juveniles as well as against those from egg and adult. rPw28CCP also showed specific reaction to the sera with active paragonimiasis. After the treatment, levels of specific antibodies in the sera gradually decreased to negative range in most patients. In some cases with persisting high antibody levels, however, the reactions at 27 kDa egg protein were sustained throughout the observation period of 34 months. The reactions at 35 and 32 kDa in adult extract and rPw28CCP disappeared rapidly after the treatment. Persistent antibody reactions even after successful treatment are provoked by continuous antigenic challenge from eggs which were not resolved by treatment. PMID:10905068
Calvopiña, Manuel; Romero, Daniel; Castañeda, Byron; Hashiguchi, Yoshihisa; Sugiyama, Hiromu
A review of national and international publications on paragonimiasis in Ecuador, epidemiological records from the Ministry of Public Health and unpublished research data was conducted to summarise the current status of the parasite/disease. The purpose of the review is to educate physicians, policy-makers and health providers on the status of the disease and to stimulate scientific investigators to conduct further research. Paragonimiasis was first diagnosed in Ecuador 94 years ago and it is endemic to both tropical and subtropical regions in 19 of 24 provinces in the Pacific Coast and Amazon regions. Paragonimus mexicanus is the only known species in the country, with the mollusc Aroapyrgus colombiensis and the crabs Moreirocarcinus emarginatus, Hypolobocera chilensis and Hypolobocera aequatorialis being the primary and secondary intermediate hosts, respectively. Recent studies found P. mexicanus metacercariae in Trichodactylus faxoni crabs of the northern Amazon. Chronic pulmonary paragonimiasis is commonly misdiagnosed and treated as tuberculosis and although studies have demonstrated the efficacy of praziquantel and triclabendazole for the treatment of human infections, neither drug is available in Ecuador. Official data recorded from 1978-2007 indicate an annual incidence of 85.5 cases throughout the 19 provinces, with an estimated 17.2% of the population at risk of infection. There are no current data on the incidence/prevalence of infection, nor is there a national control programme.
Doanh, Pham Ngoc; Guo, Zhihong; Nonaka, Nariaki; Horii, Yoichiro; Nawa, Yukifumi
Paragonimus bangkokensis and Paragonimus harinasutai, which are morphologically distinguishable species, often co-infect in the same crab intermediate hosts. Recent molecular phylogenetic analyses revealed that these two species are genetically close to each other and are considered as the sister species. While we have been studying Paragonimus adult worms obtained from the lungs of a cat experimentally infected with Paragonimus metacercariae which were morphologically identified as P. harinasutai collected from central Viet Nam, one out of 6 adult worms has grouped cuticular spines, which is a feature of P. bangkokensis. By molecular analyses, the CO1 sequence of this specimen was identical with that of P. bangkokensis, but the ITS2 and the D2 region of 28S rDNA sequences showed a two peak pattern. Then, PCR products of the ITS2 and the D2 region of 28S rDNA sequences were ligated to TOPO vector and subcloned to determine the heterozygosity. Two types of sequences were obtained from each ITS2 and D2 region of 28S; one was identical with P. harinasutai and the other with P. bangkokensis. Taking all these morphological and molecular data together, we identified this adult worm as a hybrid specimen of P. bangkokensis and P. harinasutai. Copyright © 2013 Elsevier Ireland Ltd. All rights reserved.
Doanh, P N; Hien, H V; Nonaka, N; Horii, Y; Nawa, Y
Two members of the Paragonimus skrjabini complex, P. skrjabini and P. miyazakii, are now considered as two sub-species, P. skrjabini skrjabini and P. skrjabini miyazakii. They are well known as important pathogens for human paragonimiasis in China and Japan. Recently, members of this species complex have been reported from India. Here we report the first discovery of P. skrjabini from freshwater crab hosts in Thanh Hoa province, Vietnam. For morphological and molecular phylogenetic studies, adult worms were obtained by experimental infection in cats and dogs. Molecular analyses of metacercariae and adults revealed that the P. skrjabini population from Thanh Hoa, Vietnam was almost completely identical with that from Yunnan province, China. Those populations from Thanh Hoa, Vietnam and Yunnan, China and those from Manipur, India were significantly different from P. skrjabini populations reported from other localities of China in cytochrome oxidase subunit 1 (CO1) gene sequences, indicating considerable genetic variation within the P. skrjabini complex. Moreover, low bootstrap values in the CO1 tree suggested that more variant genotypes belonging to P. skrjabini complex may be found in other Asian countries in between Vietnam and India, such as Myanmar, Laos and Thailand. Since P. skrjabini is known as a pathogen for humans, paragonimiasis cases caused by P. skrjabini might be found in Vietnam and other Asian countries.
Chen, M X; Ai, L; Zhang, R L; Xia, J J; Wang, K; Chen, S H; Zhang, Y N; Xu, M J; Li, X; Zhu, X Q; Chen, J X
In the present study, a loop-mediated isothermal amplification (LAMP) assay was developed and validated for the detection of Paragonimus westermani adults, metacercariae, and eggs in human and animal samples. The LAMP amplification can be finished in 45 min under isothermal condition at 60°C by employing a set of four species-specific primer mixtures and the results can be checked by naked-eye visualization. No amplification products were detected with deoxyribunucleic acid (DNA) of related trematode species including Fasciola hepatica, Fasciola gigantica, Clonorchis sinensis, Opisthorchis viverrini, Schistosoma mansoni, and Schistosoma japonicum. The method was further validated by examining P. westermani DNA in intermediate hosts including freshwater crabs and crayfish, as well as in sputum and pleural fluid samples from patients of paragonimiasis. These results indicated that the LAMP assay was highly specific, sensitive, and rapid, and it was approximately 100 times more sensitive than conventional specific PCR. The LAMP assay established in this study provides a rapid and sensitive tool for the detection of P. westermani DNA in freshwater crabs, crayfish, sputum, and pleural fluid samples, which has important implications for effective control of human paragonimiasis.
Shin, Myeong Heon; Kita, Hirohito; Park, Hae Young; Seoh, Ju Young
An immunoglobulin G (IgG)-coated surface, such as that found on helminth parasites, is one of the most effective physiologic stimuli for eosinophil activation. The cysteine proteases secreted by tissue-invasive helminth larvae play an important role in evasion of the immune response by degrading the host immunoglobulins. In this study, we investigated whether cysteine proteases in the excretory-secretory product (ESP) produced by Paragonimus westermani newly excysted metacercariae (PwNEM), which cause pulmonary or extrapulmonary paragonimiasis in human beings, could modify effector functions of human eosinophils stimulated with IgG. We coated 96-well plates with human IgG in the absence or presence of the ESP produced by PwNEM. When eosinophils were incubated in the wells coated with IgG in the presence of the ESP, eosinophil degranulation and superoxide production were significantly reduced compared with results for cells incubated in wells coated with IgG alone. This inhibitory effect of the ESP on IgG-induced superoxide production was dose dependent and was significantly abolished by pretreatment of the ESP with heat. These findings suggest that the cysteine proteases secreted by PwNEM attenuate both activation and degranulation of eosinophils stimulated with IgG. Thus, the cysteine proteases produced by tissue-invasive helminth larvae play crucial roles in evasion of IgG-dependent eosinophil helminthotoxicity and in reduction of eosinophil-associated tissue inflammation during the migratory period. PMID:11179333
Doanh, Pham Ngoc; Shinohara, Akio; Horii, Yoichiro; Habe, Shigehisa; Nawa, Yukifumi
Paragonimus westermani is the most well-known species among the genus Paragonimus. It is widely distributed in Asia with considerable genetic diversity to form P. westermani species complex. While P. westermani distributed in Japan, Korea, China, and Taiwan are genetically homogeneous to form the East Asia group, those found in other geographic areas are heterogeneous and would be divided into several groups. Recent discoveries of P. westermani in India and Sri Lanka highlighted new insights on molecular phylogenetic relationship of geographic isolates of this species complex. Since Vietnam is located at the east end of Southeast Asia, the intermediate position between South and East Asia, it is of interest to see whether P. westermani is distributed in this country. Here, we report that P. westermani metacercariae were found in mountainous crabs, Potamiscus sp., collected in Quangtri province in the central Vietnam. Adult worms were successfully obtained by experimental infection in cats. Molecular phylogenetic analyses revealed that P. westermani of Vietnamese isolates have high similarities with those of East Asia group.
Cho, Jun Kyong; Kwon, Hye Soo; Joo, Kyoung Hwan; Lee, Joon Sang
Changes in the expression level of splenocyte IFN-γ mRNA of Sprague-Dawley (SD) rats infected with Paragonimus westermani were analyzed by competitive reverse transcription-polymerase chain reaction (RT-PCR) followed by southern blot. The template RNA was extracted from the splenocytes of rats infected with 20 metacercariae of P. westermani. The products of competitive RT-PCR were subjected to southern blot and enhanced chemiluminescence (ECL), and analyzed with a densitometer. In comparison with that of uninfected control rat splenocytes (value of 1), the levels of mRNA expression of IFN-γ had changed to 0.747 at 1 week post infection (PI), 0.00175 at 2 week PI, 0.0217 at 3 week PI, 0.194 at 4 week PI and then to 0.537 at 5 week PI. The level at 7 week PI had returned to 1.25, comparable with that of uninfected rats. These results show that, when infected with P. westermani, the levels of IFN-γ mRNA of SD rat splenocytes were remarkably reduced by more than 500 times at 2 week PI and restored to normal level at 7 week PI. PMID:10634046
Full Text Available An epidemiological study was carried out in 2004-2005 at the anti-tuberculosis centre of Divo (Ivory Coast to collect sera from patients who consulted for tuberculosis suspicion and to estimate the seroprevalence of human paragonimosis in the context of a systematic screening. No Paragonimus egg was found in the stools and/or sputa of the 167 persons investigated. In contrast, 41 sera were ascertained with antibodies against Paragonimus africanus using ELISA testing. As the optical density (OD values related to seropositive findings were found under 0.6 (the minimal OD to detect an active paragonimosis, the above antibody titres might originate from patients in chronic or in convalescent stages, or might result of cross reactions with trematodes. Concomitantly, dissection of local crabs (Callinectes marginatus demonstrated the presence of Paragonimus metacercariae in six out of 34 examined. The parasite burdens in crabs ranged from two to 35 cysts with a mean diameter of 302 μm. In Ivory Coast, the locality of Divo must be considered an at-risk zone in reason of the presence of anti-Paragonimus antibodies in several human sera and the presence of infected crabs at the local market.
Full Text Available Stool samples from 409 pre-school and school students, living in six villages of the Cajabamba and Condebamba districts, Cajamarca, Perú, were examined using wet preparations and Lumbreras' method, looking for Paragonimus eggs. Fecal and sputum samples from two children (0.5% of 6 and 8 year-old showed eggs of Paragonimus. One hundred and twenty freshwater crabs, Hypolobocera chilensis eigenmanni, were collected from the Condebamba valley and 21 (17.5% of them were infected with P. mexicanus (syn. P. peruvianus metacercariae. Our results show the persistence of Paragonimus in human beings and in the main source of infection, the crabs.As amostras de fezes de 409 pré-escolares e escolares de 6 localidades dos distritos de Cajabamba e de Condebamba, departamento de Cajamarca, Peru, foram avaliados pelo método de Lumbreras na busca de ovos de Paragonimus sp. Dois meninos (0,5%, de 6 a 8 anos de idade, revelaram ovos de Paragonimus sp. nas suas fezes, bem como nas amostras de escarro. O exame radiográfico de tórax mostrou lesões pulmonares. Foram coletados 120 caranguejos de rio, Hypolobocera chilensis eigenmanni, no vale de Condebamba , encontrando-se 21 (17,5% infectados com metacercárias de P. mexicanus (sin. P. peruvianus. Nossos resultados revelam a persistencia de Paragonimus sp. na população na sua principal fonte de infecção, os caranguejos.
Le, Thanh H; Van De, Nguyen; Blair, David; McManus, Donald P; Kino, Hideto; Agatsuma, Takeshi
Paragonimus heterotremus Chen and Hsia (1964), and paragonimiasis caused by this species is a newly detected disease in Vietnam. Twelve samples of Paragonimus (Platyhelminthes: Trematoda: Digenea: Paragonimidae) from different life-stages (eggs, miracidia, metacercariae, adults from natural and experimental hosts) and host species (crab, dog, cat and human) were collected in different geographical locations in Vietnam. DNA sequences were obtained from each for partial mitochondrial cytochrome c oxidase subunit 1 (cox1) (387 bp) and the entire second ribosomal internal transcribed spacer (ITS-2) (361 bp). The ITS-2 sequences were identical among all specimens, including those previously reported in GenBank. For cox1, there were sequence differences between specimens from Vietnam (four provinces, different locations) and those from Guangxi (China) and Saraburi (Thailand). Phylogenetic trees inferred from cox1 and ITS-2 sequences using sequence data for 15 P. heterotremus and for other Paragonimus spp. revealed that all P. heterotremus originating from Vietnam, Thailand and China form a distinct group. This information also confirms the identity of the Vietnamese specimens as P. heterotremus.
Kim, S-H; Cai, G-B; Bae, Y-A; Lee, E-G; Lee, Y-S; Kong, Y
Phospholipid hydroperoxide glutathione peroxidase (PHGPx; GPx4) plays unique roles in the protection of cells against oxidative stress by catalysing reduction of lipid hydroperoxides. We characterized 2 novel GPx genes from a lung fluke, Paragonimus westermani (designated PwGPx1 and PwGPx2). These single copy genes spanned 6559 and 12 371 bp, respectively, and contained each of 5 intervening introns. The PwGPx2 harboured a codon for Sec and a Sec insertion sequence motif. Proteins encoded by the Paragonimus genes demonstrated a primary structure characteristic to the PHGPx family, including preservation of catalytic and glutathione-binding domains and absence of the subunit interaction domain. Expression of PwGPx1 increased gradually as the parasite matured, whereas that of PwGPx2 was temporally regulated. PwGPx2 was expressed at the basal level from the metacercariae to the 3-week-old juveniles; however, the expression was significantly induced in the 7-week-old immature worms and reached a plateau in the 12-week-old adults and eggs. PwGPx1 and PwGPx2 were largely localized in vitellocytes within vitelline glands and eggs. Oxidative stress-inducible paraquat, juglone and H2O2 substantially augmented the PwGPx1 and PwGPx2 expressions in viable worms by 1.5- to 11-fold. Our results strongly suggested that PwGPxs may actively participate in detoxification of oxidative hazards in P. westermani.
Doanh, Pham Ngoc; Horii, Yoichiro; Nawa, Yukifumi
Paragonimiasis is a food-borne parasitic zoonosis caused by infection with lung flukes of the genus Paragonimus. In Vietnam, research on Paragonimus and paragonimiasis has been conducted in northern and central regions of the country. Using a combination of morphological and molecular methods, 7 Paragonimus species, namely P. heterotremus, P. westermani, P. skrjabini, P. vietnamensis, P. proliferus, P. bangkokenis and P. harinasutai, have been identified in Vietnam. Of these, the first 3, P. heterotremus, P. westermani and P. skrjabini, are known to infect humans in other countries. However, in Vietnam, only P. heterotremus, found in some northern provinces, has been shown to infect humans. Even nowadays, local people in some northern provinces, such as Lai Chau and Yen Bai, are still suffering from P. heterotremus infection. In some provinces of central Vietnam, the prevalence and infection intensity of P. westermani metacercariae in freshwater crabs (the second intermediate hosts) are extremely high, but human cases have not been reported. Likewise, although P. skrjabini was found in Thanh Hoa Province, its pathogenicity to humans in Vietnam still remains uncertain. The results of molecular phylogenetic analyses of Vietnamese Paragonimus species provides new insights on the phylogeny and taxonomy of the genus Paragonimus. Comprehensive molecular epidemiological and geobiological studies on the genus in Vietnam and adjacent countries are needed to clarify the biodiversity and public health significance of the lung flukes.
Doanh, P N; Hien, H V; Tu, L A; Nonaka, N; Horii, Y; Nawa, Y
Conventional identification of Paragonimus species and their natural definitive hosts is based on the morphological features of adult parasites isolated from the lungs of wild mammalian hosts. However, wild animals are protected by strict regulations and sampling is not always possible. Recently, molecular techniques have been developed to identify the internal transcribed spacer (ITS) sequences of Paragonimus eggs in faeces/sputum of human patients. Also, mammalian hosts can be identified using the D-loop sequence of mitochondrial DNA in faecal samples. In this study, we used molecular techniques on faeces from wild animals collected in Da Krong Nature Reserve, Quang Tri province, central Vietnam, where Paragonimus metacercariae are highly prevalent in mountain crabs, to identify Paragonimus species and their natural definitive hosts. The results indicated that wild cats, Prionailurus bengalensis, were infected with at least three different Paragonimus species, P. westermani, P. skrjabini and P. heterotremus. Because all of these species can infect humans in Asian countries, human paragonimiasis should be considered in this area.
Moon, E-K; Lee, S-H; Goo, T W; Quan, F-S
Clonorchis sinensis and Capillaria hepatica are zoonotic parasites that mainly infect the liver and cause serious liver disorders. However, immunological parameters induced by co-infection with these parasites remain unknown. In this study, for the first time, we investigated immunological profiles induced by co-infection with C. hepatica (CH) in C. sinensis (CS)-infected rats (Sprague-Dawley). Rats were infected primarily with 50 metacercariae of C. sinensis; 4 weeks later, they were subsequently infected with 1000 infective C. hepatica eggs. Significantly higher levels of C. sinensis- or C. hepatica-specific IgG antibodies were found in the sera of rats. Interestingly, no cross-reacting antibody was observed between C. sinensis and C. hepatica infections. Significantly raised eosinophil levels were found in the blood of C. sinensis/C. hepatica co-infected rats (CS + CH) compared to the blood of rats infected singly with C. sinensis. Co-infected rats showed significantly higher levels of lymphocyte proliferation and cytokine production compared to a single C. sinensis infection. The worm burden of C. sinensis was significantly reduced in co-infected rats compared to the single C. sinensis infection. These results indicate that the eosinophils, lymphocyte proliferation and cytokine production induced by subsequent infection with C. hepatica in C. sinensis-infected rats might contribute to the observed C. sinensis worm reduction.
Lee, Dong-Hun; Kim, Ah-Ra; Lee, Su-Hwa; Quan, Fu-Shi
Human clonorchiasis, caused by the infection of Clonorchis sinensis, is one of the major health problems in Southeast Asia. However, vaccine efficacy against C. sinensis infection remains largely unknown. In this study, for the first time, we generated virus-like particles (VLPs) vaccine containing the C. sinensis tegumental protein 22.3 kDa (CsTP 22.3) and the influenza matrix protein (M1) as a core protein, and investigated the vaccine efficacy in Sprague-Dawley rats. Intranasal immunization of VLPs vaccine induced C. sinensis-specific IgG, IgG2a and IgG2c in the sera and IgA responses in the feces and intestines. Notably, upon challenge infection with C. sinensis metacercariae, significantly lower adult worm loads (70.2%) were measured in the liver of rats immunized with VLPs, compared to those of naïve rats. Furthermore, VLPs immunization induced antibody secreting cells (ASC) responses and CD4+/CD8+ T cell responses in the spleen. Our results indicated that VLPs vaccine containing C. sinensis CsTP 22.3 kDa provided partial protection against C. sisnensis infection. Thus, VLPs could be a potential vaccine candidate against C. sinensis.
Doanh, Pham N; Nawa, Yukifumi
Clonorchis sinensis and Opisthorchis viverrini are clinically important small liver flukes because of their known association with development of cholangiocarcinoma. In Vietnam, high prevalence of C. sinensis infection in humans was previously reported in northern provinces, and O. viverrini infection has been detected in several central provinces. However, diagnosis of C. sinensis and O. viverrini infections in the past was merely based on faecal egg examination. This method alone can lead to misidentification at the species level because of morphological similarity between the eggs of these liver flukes and minute intestinal trematodes of the family Heterophyidae. In fact, recent surveys in Vietnam revealed that infection with several minute intestinal flukes, such as Haplorchis pumilio and H. taichui, are much more common than infection with C. sinensis or O. viverrini, and they often co-infect humans. Thus, previously reported prevalence of small liver fluke infection in Vietnam was likely over-estimated due to mis identification of parasites in copro-parasitological examinations. In addition, there is some confusion about identification of cercariae, metacercariae and also adults of C. sinensis and O. viverrini in intermediate and definitive hosts. The aim of this review is, therefore, to draw realistic pictures of the past and present scientific reports on the epidemiology and biology of C. sinensis and Opisthorchis spp. infection in Vietnam. © The Author 2015. Published by Oxford University Press on behalf of Royal Society of Tropical Medicine and Hygiene. All rights reserved. For permissions, please e-mail: firstname.lastname@example.org.
Full Text Available Introduction. The number of humans infected by Fasciola hepatica is increasing worldwide. Humans can become accidental hosts by ingesting drinking water or plants contaminated with metacercariae. Case report. We reported a case of a 68-year-old Serbian woman, in which the diagnosis of acute fasciolosis had been established after serious diagnostic concerns. Based on clinical picture (episodic right upper quadrant abdominal pain, febrility and generalized body pain and biochemical analyses (high eosinophilia and high activity of alkaline phosphatase, she was appointed as suspected to the acute fasciolosis. Stool and duodenal aspirate exams were negative for Fasciola ova. In the absence of adequate serologic diagnostic for fasciolosis in Serbia, the diagnosis was confirmed using enzyme immunoassays and immunoblot at the Institute for Tropical Diseases in Hamburg, Germany. Soon after triclabendazole was administered, the symptoms disappeared and biochemical values returned to normal. Conclusion. The diagnosis of human fasciolosis may be problematic and delayed, especially in non endemic areas, because physicians rarely encounter this disease and a long list of other diseases must be considered in the differential diagnosis. The syndrome of eosinophilia, fever, and right upper quadrant abdominal pain suggest acute fasciolosis. Unclear source does not rule out fasciolosis.
Full Text Available Abstract Background Cathepsin cysteine proteases play multiple roles in the life cycle of parasites such as food uptake, immune invasion and pathogenesis, making them valuable targets for diagnostic assays, vaccines and drugs. The purpose of this study was to identify a cathepsin B of Clonorchis sinensis (CsCB and to investigate its diagnostic value for human helminthiases. Results The predicted amino acid sequence of the cathepsin B of C. sinensis shared 63%, 52%, 50% identity with that of Schistosoma japonicum, Homo sapiens and Fasciola hepatica, respectively. Sequence encoding proenzyme of CsCB was overexpressed in Escherichia coli. Reverse transcription PCR experiments revealed that CsCB transcribed in both adult worm and metacercaria of C. sinensis. CsCB was identified as a C. sinensis excretory/secretory product by immunoblot assay, which was consistent with immunohistochemical localization showing that CsCB was especially expressed in the intestine of C. sinensis adults. Both ELISA and western blotting analysis showed recombinant CsCB could react with human sera from clonorchiasis and other helminthiases. Conclusions Our findings revealed that secreted CsCB may play an important role in the biology of C. sinensis and could be a diagnostic candidate for helminthiases.
Fascioliasis is an important parasitic disease of grazing cattle and sheep. The experimental approach included the production of metacercariae of Fasciola hepatica from Lymnae tomentosa, infection of experimental animals, separation of fractions from whole Fasciola hepatica antigen by gel filtration on Sephadex G-100 and G-200, labelling of antigens with 125 I and examination of experimentally infected sheep and other animals for antibodies by modified radioactive single radial diffusion, indirect radioimmunodiffusion and radioactive antigen microprecipitin assay by means of direct radioimmunodiffusion and immunodiffusion using unlabelled antigen. Sufficient quantities of fluke for the production of antigen were obtained only from sheep. Infected rabbits, guinea pigs and rats yielded relatively few flukes. Labelling of the antigenic fraction can be regarded as fairly successful but the specific radioactivity of the fractions needs to be increased. Results from antibodies in infected animals by indirect radioimmunodiffusion and modified radioactive single diffusion proved not fully specific. Radioactive antigen microprecipitin assay would appear to differentiate between negative and positive sera
Moawad, M.A.; Amin, M.M.
Fasciola hepatica is the causative agent of fasciolosis in many areas in America, Europe, Africa, Asia and Australia. There is an urgent need for improved methods to control the parasite's transmission. The present study is parasitological, immunological (interleukine-1β and interleukine-6) and histopathological investigations on the immunizing effect of cross antigenicity between S. mansoni and F. hepatica against schistosomiasis and fasciolosis in mice. Parasitological study showed that vaccination with irradiated cercariae of S. mansoni or vaccination by F. hepatica whole worm extract (FhWWE) before challenged with encysted metacercariae of F. hepatica or cercariae of S. mansoni played a significant control on the parasitic infection manifested by a remarkable reduction in the means of worm count. Assessment of IL-1β and IL-6 in sera of the experimental groups showed that there are cross reactivity between fasciola / schistosoma and its relation to cross protection. Histopathological examination of vaccinated mice livers showed protection against parasite maturation and liver damage after challenged, as compared to mice infected only without vaccination
Kueakhai, Pornanan; Changklungmoa, Narin; Chaithirayanon, Kulathida; Songkoomkrong, Sineenart; Riengrojpitak, Suda; Sobhon, Prasert
A monoclonal antibody (MoAb) against recombinant Fasciola gigantica saposin-like protein 2 (rFgSAP-2) was produced by hybridoma technique using spleen cells from BALB/c mice immunized with rFgSAP-2. This MoAb is an IgG1, κ light chain isotype. By immunoblotting and indirect ELISA, the MoAb reacted specifically with rFgSAP-2, the natural FgSAP-2 at 10kDa in whole body (WB) and excretory-secretory (ES) fractions of F. gigantica. It did not cross react with antigens in WB fractions from other parasites, including Opisthorchis viverrini, Schistosoma mansoni which are human parasites, Haemonchus placei, Setaria labiato-papillosa, Eurytrema pancreaticum, Cotylophoron cotylophorum, Fischoederius cobboldi, Gigantocotyle explanatum, Gastrothylax crumenifer, and Paramphistomum cervi which are ruminant parasites. By immunohistochemistry, the FgSAP-2 protein was localized only in the cytoplasm of caecal epithelial cells of 4-week-old juvenile and adult stages, but not in metacercariae, newly excysted juvenile (NEJ), 2- and 3-week-old juveniles. This finding indicated that FgSAP-2 is an abundantly expressed parasite protein that is released into the ES, hence SAP-2 and its MoAb may be used for immunodiagnosis of ruminant and human fasciolosis. Copyright © 2012 Elsevier B.V. All rights reserved.
The cercariae of Eurytrema pancreaticum (Janson, 1889) possess four types of gland cells - proper cystogenic, penetration, ventral and dorsal gland cells. The secretion of ventral and dorsal gland cells is released into the tegument. The proper cystogenic gland cells are the largest and their contents serve for the formation of the cyst wall of metacercariae in the second intermediate host. The secretion of proper cystogenic gland cells contains besides neutral mucosubstances also acid mucosubstances with both carboxyl- and sulphogroups digestible with beta-glucuronidase. The secretion of penetration gland cells contains neutral mucosubstances and proteins with tyrosine, tryptophan and SS groups. The ventral gland cells contain mostly acid mucosubstances with sulphogroups, which are digested with beta-glucuronidase, and proteins with tyrosine, tryptophan and SH groups. The rudimentary dorsal gland cells contain a small amount of acid mucosubstances. The whole tegument of cercariae and the two main collecting canals of the excretory system exhibit a high alkaline phosphatase activity. The nerve ring and the main nerve truncs contain proteins with SH groups and hydrophilic lipids and exhibit a cholinesterase activity. The suckers contain a larger amount of glycogen.
Full Text Available Fifty specimens of Cichla piquiti were collected from the Paraná River downstream of the Ilha Solteira Hydroelectric Power Station in Brazil and surveyed for endohelminth parasites. All fish were parasitised by at least one helminth species (overall prevalence [P] = 100%. Eight parasite taxa were present: the nematode Procamallanus(Procamallanus peraccuratus in the intestines; third-stage larvae of the anisakids Contracaecumsp. and Hysterothylacium sp. in the visceral cavity, mesentery and serosa of the stomach and intestines and on the liver and spleen; the trematodes Austrodiplostomum compactum in the eye (metacercariae and Genarchella genarchella in the stomach; and the cestodes Proteocephalus macrophallus, Proteocephalus microscopicus, and Sciadocephalus megalodiscus in the intestines. Hysterothylaciumsp. larvae (P = 86% and P. microscopicus (P = 74% were the most prevalent parasites. Anisakids were more prevalent and abundant in the dry season. A negative correlation between the abundances of Hysterothylacium sp. and P. microscopicuswas observed, suggesting a competitive/antagonistic relationship between these parasites. Cichla piquiti represents a new host for four parasite species. These new records significantly increase the list of parasites of C. piquiti, contributing to the knowledge of the host-parasite relationship and the geographical distribution of these helminths.
Aksenova, Olga V; Bespalaya, Yulia V; Bolotov, Ivan N; Kondakov, Alexander V; Sokolova, Svetlana E
The strigeid digenean species Australapatemon burti (Miller, 1923) (Trematoda: Digenea: Strigeidae) was originally described from North America, but recorded in the Neotropical region (Drago et al. 2007; Hernández-Mena et al. 2014; Blasco-Costa et al. 2016) and in Central Europe (Faltýnková et al. 2007). In Europe, this species is rare, and there is not much information about its range (Faltýnková et al. 2007; Soldánová et al. 2012). Australapatemon burti has a complex life cycle with three larval stages, two of which (sporocyst and cercaria) use several species of freshwater snails, and the third stage (metacercaria) use non-specific host hirudineans (Dubois 1968; Davies & Ostrowski de Núñez 2012; Blasco-Costa et al. 2016). Adult flukes are parasitic in the intenstines of various waterfowl species, such as ducks and swans (Drago et al. 2007; Hernández-Mena et al. 2014). Currently, the molecular data on this parasite species includes only nucleotide sequences of four adult specimens from Mexico (Hernández-Mena et al. 2014). Their hosts were Mexican duck, Anas diazi Ridgway, American Wigeon, Anas americana Gmelin, Cinnamon Teal, Anas cyanoptera Vieillot, and Ruddy Duck, Oxyura jamaicensis (Gmelin) (Anserformes: Anatidae).
Phelan, Krystin; Blakeslee, April M H; Krause, Maureen; Williams, Jason D
Polychaete worms are hosts to a wide range of marine parasites; yet, studies on trematodes using these ecologically important species as intermediate hosts are lacking. During examination of the spionid polychaete Marenzelleria viridis collected on the north shore of Long Island, New York, putative trematode cysts were discovered in the body cavity of these polychaetes. In order to verify these cysts as metacercariae of trematodes, specimens of the eastern mudsnail Ilyanassa obsoleta (a very common first intermediate host of trematodes in the region) were collected for molecular comparison. DNA barcoding using cytochrome C oxidase I regions confirmed the presence of three species of trematodes (Himasthla quissetensis, Lepocreadium setiferoides, and Zoogonus lasius) in both M. viridis and I. obsoleta hosts. Brown bodies were also recovered from polychaetes, and molecular testing confirmed the presence of L. setiferoides and Z. lasius, indicating an immune response of the polychaete leading to encapsulation of the cysts. From the 125 specimens of M. viridis collected in 2014, 95 (76.8 %) were infected with trematodes; of these 95 infected polychaetes, 86 (90.5 %) contained brown bodies. This is the first confirmation that trematodes use M. viridis as a second intermediate host and that this intermediate host demonstrates a clear immune response to metacercarial infection. Future research should explore the role of these polychaetes in trematode life cycles, the effectiveness of the immune response, and transmission pathways to vertebrate definitive hosts.
Leiva, Natalia Verónica; López, Zambra; González, María Teresa; Muñoz, Gabriela
Metacercarial stages of digeneans were collected from decapod crustaceans inhabiting intertidal rocky zones in central Chile. The digeneans were identified through a molecular analysis based on the V4 region of the 18S ribosomal RNA gene. We analyzed 356 crustaceans belonging to 10 species, 115 intertidal fish belonging to 6 species, and 4 specimens of 1 coastal bird species. In total, 74.1% of crustaceans were parasitized with metacercariae. We found 1 species of Opecoelidae. This species showed low genetic divergence (0% and 0.1%) with adult digeneans found in intertidal fish and with the species Helicometrina labrisomi infesting a subtidal fish from northern Chile (Labrisomus philippii). Additionally, we found 2 species of Microphallidae, 1 closely related to Maritrema (1.3% genetic distance) and the other related to Microphallus (5% genetic distance). Therefore, our findings showed that the decapod crustaceans are relevant hosts in food webs from the southeastern Pacific coast. Furthermore, we found 5 species of crustaceans as second intermediate hosts for H. labrisomi and 2 species as secondary intermediate hosts for 2 Microphallidae, which contribute to elucidate parts of their life cycles through molecular markers and extended the host distribution of H. labrisomi in the southeastern Pacific coast.
Kanev, I.; Sorensen, R.; Sterner, M.; Cole, Rebecca A.; Fried, B.
The life cycle of Echinoparyphium rubrum (Cort, 1914) comb. n. has been completed experimentally. All of the developmental stages egg, miracidium, sporocyst, mother and daughter rediae, cercaria, metacercaria, and adult were examined and described. The miracidia infected freshwater snails of the genus Physa , P. gyrina and P. occidentalis. Attempts to infect snails of the genera Lymnaea, L. auricularis, L. peregra, L. truncatula and Bulinus, B. truncatus failed. Cercariae infected various pulmonate and prosobranch freshwater snails, mussels, frogs, water turtles and planarians. The adults developed in the small intestine of birds and mammals. The identity and major characteristics of Echinoparyphium rubrum are discussed. Synonyms of E. rubrum are Cercaria rubra Cort, 1914; Cercaria biflexa Faust, 1917; Cercaria chisolenata Faust, 1918; Echinostoma callawayensis Barker et Noll, 1915; Echinostoma revolutum of Johnson (1920); Echinoparyphium elegans of Cannon (1938), of Bain and Trelfall (1977), of Mahoney and Trelfall (1977); and Echinoparyphium recurvatum of Jilek (1977), Harley (1972), Sankurathri and Holmes (1976). Comparisons are made between E. rubrum and its 43-collar-spined allies: E. flexum from North America, E. cinctum from Europe, E. dunni from Asia and E. elegans from Africa.
Beck, Melissa A; Goater, Cameron P; Colwell, Douglas D
Epidemiological rate parameters of host generalist parasites are difficult to estimate, especially in cases where variation in parasite performance can be attributed to host species. Such cases are likely common for generalist parasites of sympatric grazing mammals. In this study, we combined data from experimental exposures in cattle and sheep and natural infections in elk to compare the recruitment, morphology and reproduction of adult Dicrocoelium dendriticum, a generalist trematode that has emerged in sympatric grazing hosts in Cypress Hills Provincial Park, Alberta. Overall, there were no significant differences in the recruitment of metacercariae and in the pre-patency period of adults in experimentally exposed cattle and sheep. All flukes reached reproductive maturity and the degree of reproductive inequality between individual flukes within each infrapopulation was moderate and approximately equal among the three host species. Neither fluke size nor per capita fecundity was constrained by density dependence. Thus, fitness parameters associated with growth and reproduction were approximately equivalent among at least three species of definitive host, two of which are sympatric on pastures in this Park. The generalist life-history strategy of this trematode, which is known to extend to other stages of its life cycle, has likely contributed to its invasion history outside its native range in Europe.
Segade, P; García-Estévez, J; Arias, C; Iglesias, R
Heliciculture is an excellent alternative to obtain edible snails but its viability is seriously threatened by pathogens. A parasitological survey was conducted in 3 mixed system-based heliciculture farms in Galicia (NW Spain), with the species Tetrahymena rostrata, Tetrahymena limacis, Tetratrichomonas limacis, Cryptobia helicogenae, Brachylaima aspersae (metacercariae and sporocysts), Alloionema appendiculatum, Nemhelix bakeri, and Riccardoella limacum being commonly found infecting Helix aspersa aspersa (petit-gris) snails. With the exception of C. helicogenae, N. bakeri, and B. aspersae sporocysts, all species were also detected in Helix aspersa maxima (gros-gris) snails, although generally with lower parameters. Most monoxenous infections, and consequently multiple parasitism, exhibited a rising trend during the first 2 months of intensive mating, with tendencies being slowed down or even reversed during the third month as a result of accumulated mortality and a sampling-derived reduction in host density. No parasites were vertically transmitted and infections were initially acquired from invading gastropod and micromammal reservoirs during fattening. Finally, artificial hibernation reduced significantly the prevalence of most species. These results confirm the importance of parasites in heliciculture and emphasize the need to prevent the entry of wild reservoirs into the farms and to rapidly remove the carcasses of dead snails from the reproduction units and fattening pens.
Yamada, F H; Santos, L N; Takemoto, R M
The gills of 41 Cichla piquiti and 39 C. kelberi from Itaipu and Lajes reservoirs, respectively, Brazil, were examined to describe the ectoparasite assemblages of these two non-native peacock-bass populations. All ectoparasite species of the two studied hosts (C. piquiti and C. kelberi) were dominant, but Ascocotyle sp. (metacercariae) was the prevalent (58.53%) and most abundant helminth species in C. piquiti hosts, while Sciadicleithrum ergensi was the dominant species in C. kelberi hosts. Gill ectoparasites of C. piquiti and C. kelberi showed a typical pattern of overdispersion or aggregation, which is commonly reported for many other freshwater fishes. Ectoparasite prevalence and abundance did not vary between host sexes of the two Cichla populations. The prevalence and abundance of Ascocotyle sp. were positively correlated with C. piquiti standard length (SL), but only the abundance of S. ergensi showed a positive correlation with C. kelberi SL. Although environmental differences between reservoirs might also have influenced the results, we anticipated that the presence of a close congener in Itaipu reservoir and the lack of other Cichla species in Lajes reservoir were the key factors to explain the contrasts between C. piquiti and C. kelberi gill ectoparasites. Overall, our results suggest that the trend of parasite species loss through the invasion process may have contributed to the establishment of non-native C. piquiti and C. kelberi populations in Brazilian reservoirs.
Alda, Pilar; Bonel, Nicolás; Cazzaniga, Néstor J.; Martorelli, Sergio R.
The aim of this work was to evaluate the effects of parasitism and certain environmental factors on the shell size of Heleobia australis (Hydrobiidae, Cochliopinae). We report sporocysts and metacercariae of Microphallus simillimus (Microphallidae, Trematoda) parasitizing the gonad and digestive gland of H. australis specimens from two sites of Bahía Blanca estuary, Argentina. The prevalence of infection was significantly higher (34.17% in winter and 68.14% in late spring) in snails from the outer estuary at Site 2 than in those from the inner estuary at Site 1 (5.88% and 4.71% respectively). The only known definitive host for this digenean is the white-backed stilt Himantopus melanurus (Recurvirostridae, Aves), most abundant in the estuary during winter. Parasitism by M. simillimus causes variations in the shell dimensions of H. australis, the shells of infected snails being narrower than those of uninfected snails. Snails from Site 2 were found in general to be significantly smaller than those at Site 1, possibly as a result of differences in environmental factors such as the degree of exposure to wave energy, the allocation of energy to reproduction rather than growth (induced by predation and/or parasitic castrators) and anthropogenic stressors.
Greiman, Stephen E; Tkach, Vasyl V
Bacteria of the genus Neorickettsia are obligate intracellular endosymbionts of parasitic flukes (Digenea) and are passed through the entire complex life cycle of the parasite by vertical transmission. Several species of Neorickettsia are known to cause diseases in domestic animals, wildlife, and humans. Quantitative data on the transmission of the bacteria through the digenean life cycle is almost completely lacking. This study quantified for the first time the abundance of Neorickettsia within multiple stages of the life cycle of the digenean Plagiorchis elegans. Snails Lymnaea stagnalis collected from a pond in North Dakota were screened for the presence of digenean cercariae, which were subsequently tested for the presence of Neorickettsia. Three L. stagnalis were found shedding P. elegans cercariae infected with Neorickettsia. These snails were used to initiate three separate laboratory life cycles and obtain all life cycle stages for bacterial quantification. A quantitative real-time PCR assay targeting the GroEL gene was developed to enumerate Neorickettsia sp. within different stages of the digenean life cycle. The number of bacteria significantly increased throughout all stages, from eggs to adults. The two largest increases in number of bacteria occurred during the period from eggs to cercariae and from 6-day metacercariae to 48-h juvenile worms. These two periods seem to be the most important for Neorickettsia propagation through the complex digenean life cycle and maturation in the definitive host.
MacLeod, C D; Poulin, R
Ocean acidification is predicted to cause major changes in marine ecosystem structure and function over the next century, as species-specific tolerances to acidified seawater may alter previously stable relationships between coexisting organisms. Such differential tolerances could affect marine host-parasite associations, as either host or parasite may prove more susceptible to the stressors associated with ocean acidification. Despite their important role in many ecological processes, parasites have not been studied in the context of ocean acidification. We tested the effects of low pH seawater on the cercariae and, where possible, the metacercariae of four species of marine trematode parasite. Acidified seawater (pH 7.6 and 7.4, 12.5 °C) caused a 40-60% reduction in cercarial longevity and a 0-78% reduction in metacercarial survival. However, the reduction in longevity and survival varied distinctly between parasite taxa, indicating that the effects of reduced pH may be species-specific. These results suggest that ocean acidification has the potential to reduce the transmission success of many trematode species, decrease parasite abundance and alter the fundamental regulatory role of multi-host parasites in marine ecosystems. Copyright © 2015 Australian Society for Parasitology Inc. Published by Elsevier Ltd. All rights reserved.
Full Text Available Clonorchis sinensis and Opisthorchis viverrini are both important fish-borne pathogens, causing serious public health problem in Asia. The present study developed an assay integrating real-time PCR and high resolution melting (HRM analysis for the specific detection and rapid identification of C. sinensis and O. viverrini. Primers targeting COX1 gene were highly specific for these liver flukes, as evidenced by the negative amplification of closely related trematodes. Assays using genomic DNA extracted from the two flukes yielded specific amplification and their identity was confirmed by sequencing, having the accuracy of 100% in reference to conventional methods. The assay was proved to be highly sensitive with a detection limit below 1 pg of purified genomic DNA, 5 EPG, or 1 metacercaria of C. sinensis. Moreover, C. sinensis and O. viverrini were able to be differentiated by their HRM profiles. The method can reduce labor of microscopic examination and the contamination of agarose electrophoresis. Moreover, it can differentiate these two flukes which are difficult to be distinguished using other methods. The established method provides an alternative tool for rapid, simple, and duplex detection of C. sinensis and O. viverrini.
Marcos Tavares Dias
Full Text Available This paper describes the first study on parasite diversity in Auchenipterus nuchalis Spix & Agassiz, 1829 (Auchenipteridae. In 31 fish caught in a tributary of the Amazon River, 10,708 parasites were collected, such as Ichthyophthirius multifiliis, Piscinoodinium pilullare, Cosmetocleithrum striatuli, metacercariae of Posthodiplostomum sp, and larvae and adults of Procamallanus (Spirocamallanus inopinatus. These parasite species showed aggregated dispersion, except for C. striatuli, which had a uniform dispersion. The component community of parasites showed a low Brillouin diversity (0.67 ± 0.27, low species richness (3.5 ± 0.8 and low evenness (0.43 ± 0.17, and it was characterized by the presence of species with high prevalence and abundance. Protozoan species were the prevalent parasites, which may be a consequence of the host’s mode of life, while the low presence of endoparasites in A. nuchalis suggests that this fish may occupy an intermediate trophic level in the food web. These data represent the first record of such parasite species for this host.
Maria Danielle Figueiredo Guimarães Hoshino
Full Text Available The present study provides the first investigation on the ecological aspects of the parasites M. lippincottianusfrom the Amazonian basin, as well as the parasite-host relationship. 76 out of the examined fish (98.7% were parasitized by at least one species of parasites. A total of 8,774 parasites were collected, being Ichthyophthirius multifiliis, Anacanthorus jegui, Dadayius pacupeva, Digenea gen. sp. (metacercariae, Procamallanus (Spirocamallanus inopinatus, Procamallanus (Spirocamallanus sp., Spinoxyuris oxydoras, Contracaecum sp. larvae, Dolops longicauda and Hirudinea gen. sp. Ichthyophthirius multifiliis was the dominant species, followed by A. jegui. Among the endoparasites, S. oxydoras and D. pacupeva were predominant. The mean diversity of parasites was HB = 0.96 ± 0.32 and there was aggregate distribution pattern. A positive correlation of body weight with the abundance of I. multifiliis and S. Oxydoras was observed, whereas a negative correlation of body weight with abundance of the Contracaecum sp. larvae was found. The relative condition factor (Kn was not negatively affected by parasites, and a positive correlation between Kn and abundance of I. multifiliis, S. oxydoras and D. pacupeva was found. This study is the first one to record I. multifiliis, D. longicauda and A. jegui parasitizing M. lippincottianus, as well as the first record of D. pacupeva and S. oxydoras in the Amazonas river system.
Mbokane E. M.
Full Text Available This study provides information on seasonal occurrence of developmental stages of endoparasites infecting three cyprinids in the Nwanedi-Luphephe dams, Limpopo River System. Labeobarbus marequensis (Smith, 1841, Barbus trimaculatus Peters, 1852 and Barbus radiatus Peters, 1853 were investigated seasonally from January 2008 to October 2008. The following larvae of metazoan parasites were collected: Diplostomum sp. from the eyes of L. marequensis and B. trimaculatus; Ornithodiplostomum sp. from the gills of B. trimaculatus; Posthodiplostomum sp. from muscle, skin and fins of B. trimaculatus and B. radiatus; third-stage Contracaecum larvae (L3 from the mesentery fats and on the liver lobes of L. marequensis and B. trimaculatus and gryporynchid cestode larvae from the outer intestinal wall of B. radiatus. All the flukes encountered were metacercariae. Diplostomum sp. and Contracaecum sp. dominated the parasite communities. Their prevalence exhibited seasonal fluctuations with maxima in summer. Factors likely to influence fish infection such as the body size of fish and their condition factors were also briefly considered in this study.
Bolbophorus damnificus n. sp. (Digenea: Bolbophoridae) from the channel catfish Ictalurus punctatus and American white pelican Pelecanus erythrorhynchos in the USA based on life-cycle and molecular data.
Overstreet, Robin M; Curran, Stephen S; Pote, Linda M; King, D Tommy; Blend, Charles K; Grater, Walter D
The common pathogenic prodiplostomulum metacercaria in the flesh, mostly near the skin, of pond-produced channel catfish Ictalurus punctatus has been demonstrated to be Bolbophorus damnificus Overstreet & Curran n. sp. The catfish acquires the infection from the snail Planorbella trivolvis, the only known first intermediate host, and the species is perpetuated through the American white pelican Pelecanus erythrorhynchos, as confirmed by experimental infections with nestling and dewormed adult pelican specimens in conjunction with molecular data. It differs from the cryptic species Bolbophorus sp., also found concurrently in the American white pelican, by having eggs 123-129 microm rather than 100-112 microm long and consistent low values for nucleotide percentage sequence similarity comparing COI, ITS 1/2, 18S rRNA and 28S rRNA fragments. Bolbophorus sp. is comparable but most likely distinct from B. confusus (Kraus, 1914), which occurs in Europe and has eggs 90-102 microm long. Its intermediate hosts were not demonstrated. The adults of neither of the confirmed North American species of Bolbophorus were encountered in any bird other than a pelican, although several shore birds feed on infected catfish, and B. damnificus can survive but not mature when protected in the mouse abdominal cavity. B. ictaluri (Haderlie, 1953) Overstreet & Curran n. comb., a species different from B. damnificus, is considered a species inquirenda.
Full Text Available Background: The purpose of this study was to perform seroepidemiological investigation for determining the status of human fasciolosis in Pirabad Village, Lorestan Province, western Iran. Methods: Blood samples were taken from residents of the village including 801 individuals. Sera were separated and stored at -20°C until used. The samples were analyzed using ELISA. Results: Anti-Fasciola antibodies were detected in 6 (0.7% individuals. Difference between age, sex and drinking or swimming in the surface water with seropositivity to fasciolosis was not significant. Out of 7 shepherds, 1 (14.3% was seropositive. Due to the small number of shepherds, comprehensive statistical inference in this regard cannot be done. Significant difference was detected between seropositivity to fasciolosis and consuming local freshwater vegetables during the last 6 months (P=0.001.Conclusion: Metacercariae carrying local freshwater plants might be the main source of contamination because consumption of these kinds of vegetables was confirmed by all participants. Awareness of local communities regarding the danger of freshwater plant consumption, through health education programs, will decrease the risk of infection.
Jeandron, Aurelie; Rinaldi, Laura; Abdyldaieva, Gulnara; Usubalieva, Jumagul; Steinmann, Peter; Cringoli, Giuseppe; Utzinger, Jürg
Dicrocoelium dendriticum is the causative agent of a rare food-borne zoonosis of the human biliary tract, dicrocoeliasis, for which few human prevalence data are available. Infection occurs through the ingestion of ants containing metacercariae, whereas pseudo-infections (presence of D. dendriticum eggs in stool in the absence of adult worms) are due to the consumption of infected animal liver. Here, results from a cross-sectional survey carried out among 138 children aged 2-15 yr in a peri-urban area of Kyrgyzstan are reported. Each child provided 1 stool sample that was subjected to the FLOTAC technique. Eggs of D. dendriticum were diagnosed in 11 children (prevalence 8.0%; 95% confidence interval 4.5-13.7%). Although no distinction could be made between true and pseudo-infections, the prevailing animal husbandry system and the diet and hygienic conditions of the study area suggest that the social-ecological system in Kyrgyzstan is conducive for human transmission of D. dendriticum. There is a need to investigate the epidemiology of dicrocoeliasis in Kyrgyzstan, placing emphasis on the distinction between true and pseudo-infections.
Alcântara, Natália Milhomem; Tavares-Dias, Marcos
This study compared the parasite communities of Hoplias malabaricus and Hoplerythrinus unitaeniatus from Amazon river system. Hoplias malabaricus were infected by Ichthyophthirius multifiliis, Piscinoodinium pillulare, Tetrahymena sp., Urocleidoides eremitus, Braga patagonica, metacercariae of Clinostomum marginatum, Procamallanus (Spirocamallanus) inopinatus, larvae of Contracaecum sp. and larvae of Nomimoscolex matogrossensis. Hoplerythrinus unitaeniatus were also infected by these same species of protozoans, nematodes, digeneans and cestodes, except for Tetrahymena sp. and B. patagonica, which were replaced by Argulus pestifer, Urocleidoides sp., Whittingtonocotyle caetei, Whittingtonocotyle jeju and Gorytocephalus spectabilis. For both hosts, I. multifiliis and P. pillulare were the predominant parasites. Most of the parasites presented an overdispersion. Parasite species richness, Brillouin diversity, evenness and Berger-Parker dominance were similar for the two hosts. The length and weight of H. malabaricus showed a positive correlation with the abundance of U. eremitus and Contracaecum sp., while the weight of H. unitaeniatus showed a positive correlation with the abundance of I. multifiliis. The diversity of ectoparasites seemed to be influenced by the behavior of these two hosts. This was shown by the similar parasite communities and was characterized by low species diversity, low evenness and low richness, and by a high prevalence of ectoparasites.
Alina Huiza F.
Full Text Available Se determina la presencia de Fasciola hepatica L., en la localidad de Huinco, zona no ganadera de la Provincia de Huarochirí, Departamento de Lima, setiembre 1971 - julio de 1972. Se examinaron caracoles Lymnaea viator y Physa venustula en busco de estadios evolutivos de F. hepatica, así como exámenes de heces de la población humana en busca de huevos del parásito. Se encontraron caracoles L. viator infectados con estadios larvarios de Fasciola, lo cual se comprobó al desarrollarse ejemplares adultos del parásito en las vías biliares de ratones que fueron infectados con metacercarias obtenidas de los caracoles. La infección humana por Fasciola hepatica fue bajo (0.88%. No existiendo ganadería en la localidad, se concluye que la infección del caracol ocurre por lo llegada a Huinco de huevos de F. hepatica por los cursos de agua de localidades ganaderos situadas en zonas más altos que Huinco.
Zintl, Annetta; Talavera, Silvia; Sacchi-Nestor, Carlotta; Ryan, Marion; Chryssafidis, Andreas; Mulcahy, Grace
Fasciola hepatica is a common and economically important parasite of sheep and cattle. Although its marked genetic heterogeneity is well recognised, an association between haplotypes and specific phenotypic traits has yet to be identified. Using experimental infections in cattle this study investigated whether a fragment of mitochondrial DNA (coding for cytochrome c oxidase subunit III, transfer RNA histidine and cytochrome b) and 3 nuclear microsatellite loci (Fh15, Fh23 and Fh25) could be used as markers for the parasite's ability to complete its tissue migration and establish in the liver of the final host. While we did not detect any shift in the frequency of the various genotypes in the population of metacercariae used for the infection on the one hand and the flukes collected from the liver on the other, there was an indication that parasites with heterozygous microsatellite alleles may have a selective advantage over homozygote parasites during their migration in the final host. Copyright © 2015 Elsevier B.V. All rights reserved.
Garcia-Campos, Andres; Baird, Alan W; Mulcahy, Grace
Fasciola hepatica is a parasitic trematode that causes serious losses to livestock producers, and also zoonotic disease. The limitations of chemotherapy for the control of fasciolosis have led to significant interest in the development of vaccines to protect cattle and sheep from infection. However, relatively few studies have concentrated on the mechanisms of invasion of the gut by newly excysted juvenile liver flukes (NEJ) and the host response triggered by this event. The aim of this work was to develop an in vitro model to study invasion by NEJ, while also reducing the requirement for challenge infections of experimental animals. Fasciola hepatica metacercariae were excysted in vitro and placed into compartments containing rat distal jejunal sheets. Variations in incubation medium, chamber size and incubation temperature were used to identify optimal conditions for NEJ migration across the gut. Histological examination showed increased migration until 120 min post-incubation. The use of RPMI, without gassing at 39 °C, as the incubation medium was found to be optimal, with 40·5% of NEJ migrating after 150 min. This study describes a readily-reproducible method for studying the migration of F. hepatica NEJ within the definitive host. It will be useful for identifying potential drug and vaccine targets.
Kołodziejczyk, Lidia; Podraza, Wojciech; Gonet, Bolesław; Dzika, Ewa; Kosik-Bogacka, Danuta I
The aim of this study was to determine the effect of an extremely low-frequency magnetic field (ELFMF) on the production of liver fluke larvae in a parasite-host system: Fasciola hepatica--Galba truncatula. Both F. hepatica eggs and F. hepatica-infected snails were exposed to an ELFMF (50 Hz, 2.0 mT) for 14 days and 36 days, respectively. F. hepatica-infected snails were divided into 4 groups, 10 specimens each. The snails of groups I and II were infected with F. hepatica larvae--miracidia obtained from control cultures, while the snails of groups III and IV were infected with miracidia reared from eggs that had been incubated in an ELFMF. After infection, the snails of groups II and IV were placed in an ELFMF, while those of groups I (control) and III were housed outside the ELFMF. At 36 days post-infection (dpi) there were no statistically significant differences between the number of F. hepatica larvae--cercariae and metacercariae, obtained from G. truncatula snails in the control group (group I) and the snail groups exposed to ELFMF (groups II, III and IV). However, a statistically significant difference between the average number of F. hepatica larvae in snail groups III and IV may indicate that the duration of exposure to ELFMF, i.e. embryogenesis period vs. the entire larval development, played a role in the production of F. hepatica larvae, and resulted in a reduction of their number.
Silveira, Tony; Remião, Mariana H; Robaldo, Ricardo B; Freitas, Kaio; Valente, Ana Luísa S
The distribution of franciscana Pontoporia blainvillei Gervais & d'Orbigny, 1884, is restricted to the estuarine and coastal waters of the southwestern Atlantic. The diet of this dolphin is composed of fishes, squids, and shrimps, many of which harbor helminths that may infect franciscana. Larval forms of the trematode Synthesium pontoporiae and the acanthocephalan Bolbosoma turbinella have been recorded in franciscana; however, they have not yet been identified in any of the prey species of this cetacean. We evaluated 3 components of the diet of franciscana as possible transmission sources of parasitiasis. Specimens of São Paulo squid Doryteuthis sanpaulensis (n = 50), banded croaker Paralonchurus brasiliensis (n = 43), and rough scad Trachurus lathami (n = 50) were necropsied. Organs were washed and examined under a stereomicroscope. Helminths were collected and mounted on slides. None of the species analyzed showed infection by metacercariae of S. pontoporiae. Helminths found in São Paulo squid have not been recorded in franciscana. Cistacanths of Corynosoma australe were found in the coelomic cavity and mesentery of croaker (prevalence [P] = 53.49%; mean infection intensity [MII] = 6.74) and scad (P = 4%; MII = 1.50). Cistacanths of B. turbinella were also found in the same sites in scad (P = 14%; MII = 2.14). Banded croaker and rough scad are recorded in this study as new paratenic hosts for C. australe, while scad is a new paratenic host for B. turbinella.
Full Text Available This contribution reports the parasites found in the mangrove mussel Mytella guyanensis in Camamu Bay, Bahia, Brazil. Samples were collected monthly from September 2006 through October 2007. A total of 460 individuals were collected, fixed in Davidson's solution, and processed by standard histological techniques, and the sections were stained with Harris hematoxylin and eosin (H&E. The water temperature ranged from 23.5 to 31.6 ºC, and the salinity from 25 to 37‰. Microscopic analysis showed Rickettsia-like organisms (RLOs, Nematopsis sp. (Apicomplexa, and Platyhelminthes, including a turbellarian, sporocysts of Bucephalus sp., metacercariae, and metacestodes of Tylocephalum sp. Parasites were observed mainly in the gills, mantle, and digestive gland. The prevalence of Nematopsis sp. was 100%, and in heavily infected mussels the tissues of the labial palps were damaged. RLOs occurred in high prevalence and intensity of infection in some periods. The digenean sporocysts showed moderate prevalence but high intensity of infection, and caused parasitic castration. In general, there was no significant spatial or temporal variation (p > 0.05 of the parasites, which is probably attributable to the small variations of temperature and salinity in the region.
Claudia Silveira São Sabas
Full Text Available The parasite fauna of catfish, Pimelodus pohli, from the São Francisco River Basin is presented. A total of 45 catfish from the upper São Francisco River (45°15′44″W 18°13′25″S, were examined from July 2009 to September 2011. Forty-three catfish (95.5% were infected by at least one parasite species, with 885 parasite specimens being found, distributed across 17 species: Monogenea (Demidospermus uncusvalidus, Pavanelliella pavanellii, and Scleroductus sp.; Eucestoda (plerocercoids of Proteocephalidea; Digenea (metacercariae of Austrodiplostomum compactum, adults of Auriculostoma platense and Kalipharynx sp., and juvenile of Prosthenhystera obesa; Nematoda (larvae of Contracaecum sp., Hysterothylacium sp., Procamallanus pimelodus, Procamallanus sp., and unidentified of Cucullanidae, and adults of Cucullanus caballeroi, Philometra sp., and Procamallanus freitasi; and Acanthocephala (adults of Neoechinorhynchus pimelodi. Procamallanus freitasi and Scleroductus sp. were the taxa with the highest prevalence. Demidospermus uncusvalidus, P. freitasi, and Scleroductus sp. were the dominant species. The host's sex did not influence parasitic indexes; however, the total length of the catfish did appear to have some influence. The parasites, with except for P. obesa, were registered for the first time in P. pohli, as well as the occurrence of Kalipharynx sp. and C. caballeroi among pimelodid hosts from São Francisco River and South America.
Sabas, Claudia Silveira São; Brasil-Sato, Marilia Carvalho
The parasite fauna of catfish, Pimelodus pohli, from the São Francisco River Basin is presented. A total of 45 catfish from the upper São Francisco River (45°15'44″W 18°13'25″S), were examined from July 2009 to September 2011. Forty-three catfish (95.5%) were infected by at least one parasite species, with 885 parasite specimens being found, distributed across 17 species: Monogenea (Demidospermus uncusvalidus, Pavanelliella pavanellii, and Scleroductus sp.); Eucestoda (plerocercoids of Proteocephalidea); Digenea (metacercariae of Austrodiplostomum compactum, adults of Auriculostoma platense and Kalipharynx sp., and juvenile of Prosthenhystera obesa); Nematoda (larvae of Contracaecum sp., Hysterothylacium sp., Procamallanus pimelodus, Procamallanus sp., and unidentified of Cucullanidae, and adults of Cucullanus caballeroi, Philometra sp., and Procamallanus freitasi); and Acanthocephala (adults of Neoechinorhynchus pimelodi). Procamallanus freitasi and Scleroductus sp. were the taxa with the highest prevalence. Demidospermus uncusvalidus, P. freitasi, and Scleroductus sp. were the dominant species. The host's sex did not influence parasitic indexes; however, the total length of the catfish did appear to have some influence. The parasites, with except for P. obesa, were registered for the first time in P. pohli, as well as the occurrence of Kalipharynx sp. and C. caballeroi among pimelodid hosts from São Francisco River and South America.
Santos-Clapp, Michelle Daniele; Brasil-Sato, Marilia Carvalho
A total of 112 specimens of yellow peacock bass, Cichla kelberi, were collected in the Três Marias Reservoir, upper São Francisco River, State of Minas Gerais, between 2004 and 2005. Twelve species of parasites were collected: two species of ectoparasites (the monogenetic Gussevia sp. and the copepod Ergasilus sp.); and ten species of endoparasites, including four species of adult endoparasites: the cestodes Proteocephalus macrophallus and P. microscopicus, and the nematodes: Capillostrongyloides sentinosa and Rhabdochona acuminata, and six species of larval endoparasites: the metacercariae of Austrodiplostomum compactum and the nematodes: Hysterothylacium sp., Contracaecum sp. Type 1, Procamallanus (Spirocamallanus) inopinatus, Spiroxys sp., and Goezia sp.. A significant positive correlation in the abundance of the two adult endoparasites species, P. macrophallus and P. microscopicus, may indicate that such associated species had the same ecological requirements. The positive correlations between the prevalence of the larval endoparasites, Hysterothylacium sp. and A. compactum, may indicate that there was opportunism for the two generalists species. Ergasilus sp., C. sentinosa, R. acuminata and Hysterothylacium sp. were recorded for the first time in C. kelberi, and this increased their list of known hosts. In addition, both proteocephalid species and C. sentinosa can extend their known biogeographical distribution to the Três Marias Reservoir.
Franceschini, Lidiane; Zago, Aline Cristina; Zocoller-Seno, Maria Conceição; Veríssimo-Silveira, Rosicleire; Ninhaus-Silveira, Alexandre; da Silva, Reinaldo José
Fifty specimens of Cichla piquiti were collected from the Paraná River downstream of the Ilha Solteira Hydroelectric Power Station in Brazil and surveyed for endohelminth parasites. All fish were parasitised by at least one helminth species (overall prevalence [P] = 100%). Eight parasite taxa were present: the nematode Procamallanus (Procamallanus) peraccuratus in the intestines; third-stage larvae of the anisakids Contracaecum sp. and Hysterothylacium sp. in the visceral cavity, mesentery and serosa of the stomach and intestines and on the liver and spleen; the trematodes Austrodiplostomum compactum in the eye (metacercariae) and Genarchella genarchella in the stomach; and the cestodes Proteocephalus macrophallus, Proteocephalus microscopicus, and Sciadocephalus megalodiscus in the intestines. Hysterothylacium sp. larvae (P = 86%) and P. microscopicus (P = 74%) were the most prevalent parasites. Anisakids were more prevalent and abundant in the dry season. A negative correlation between the abundances of Hysterothylacium sp. and P. microscopicus was observed, suggesting a competitive/antagonistic relationship between these parasites. Cichla piquiti represents a new host for four parasite species. These new records significantly increase the list of parasites of C. piquiti, contributing to the knowledge of the host-parasite relationship and the geographical distribution of these helminths.
Brasil-Sato, Marilia de C; Dos Santos, Michelle D
Specimens of Conorhynchos conirostris (Valenciennes, 1840) (Osteichthyes: Siluriformes) were collected from the upper São Francisco River (18 degrees 12'32''S, 45 degrees 15'41''W) in the municipality of Três Marias, Minas Gerais, Brazil, to investigate their parasitofauna. Of the 24 pirá fish collected, 8 were male (33.3%) and 16 were female (66.7%). Of this total, 12 were parasitized (50%). Nine species of parasites were found: Helobdella sp., Creptotrema creptotrema Travassos, Artigas & Pereira, 1928; Palaeocryptogonimus claviformis Szidat, 1954; metacercariae of Clinostomum sp. and of Austrodiplostomum compactum (Lutz 1928); Procamallanus (Spirocamallanus) sp. (young specimen); larvae of Anisakidae and Rhabdochona sp.; and Neoechinorhynchus sp. (young specimen). Helobdella sp. had the highest prevalence, followed by C. creptotrema (most abundant) and P. claviformis. Creptotrema creptotrema was dominant species in the parasite community of C. conirostris. There was no influence of the host size and sex on the prevalence and abundance of parasites, with P. claviformis being found only in male hosts. With the exception of Procamallanus (Spirocamallanus) sp., this was the first record of these parasites in C. conirostris.
Natália Milhomem Alcântara
Full Text Available This study compared the parasite communities of Hoplias malabaricus and Hoplerythrinus unitaeniatus from Amazon river system. Hoplias malabaricus were infected by Ichthyophthirius multifiliis, Piscinoodinium pillulare, Tetrahymena sp., Urocleidoides eremitus, Braga patagonica, metacercariae of Clinostomum marginatum, Procamallanus(Spirocamallanus inopinatus, larvae of Contracaecum sp. and larvae of Nomimoscolex matogrossensis. Hoplerythrinus unitaeniatus were also infected by these same species of protozoans, nematodes, digeneans and cestodes, except for Tetrahymena sp. and B. patagonica, which were replaced by Argulus pestifer, Urocleidoides sp., Whittingtonocotylecaetei, Whittingtonocotyle jeju and Gorytocephalus spectabilis. For both hosts, I. multifiliis and P. pillulare were the predominant parasites. Most of the parasites presented an overdispersion. Parasite species richness, Brillouin diversity, evenness and Berger-Parker dominance were similar for the two hosts. The length and weight of H. malabaricusshowed a positive correlation with the abundance of U. eremitusand Contracaecum sp., while the weight of H. unitaeniatus showed a positive correlation with the abundance of I. multifiliis. The diversity of ectoparasites seemed to be influenced by the behavior of these two hosts. This was shown by the similar parasite communities and was characterized by low species diversity, low evenness and low richness, and by a high prevalence of ectoparasites.
Acosta, A A; Queiroz, J; Brandão, H; Silva, R J
This study assessed the helminth fauna of Astyanax fasciatus in two distinct sites of the Taquari River, São Paulo State, with 30 individuals sampled in a lotic site and 30 in a lentic site, recording the monogeneans: Cacatuocotyle paranaensis, Characithecium costaricensis, Diaphorocleidus kabatai, Jainus sp., Notozothecium sp. and Gyrodactylus sp., the digenean Antorchis lintoni and no-identified metacercariae; the nematode Procamallanus (Spirocamallanus) inopinatus and no-identified larvae. The mean abundances of total monogeneans (U = 1053; p = 0.042) and C. costaricensis (U = 1107; p = 0.005) were higher in the lotic site. This difference may be due to the higher density of the host population in the lotic site, and the water transparence in lentic environments that prevents A. fasciatus to form shoals, precluding the exchange of parasites with direct cycle within a host population. This study is the first report of the helminth fauna of A. fasciatus in the Taquari River, with ten taxa recorded, and reports A. fasciatus as a new host for Notozothecium sp. and C. paranaensis.
de Oliveira Ferreira, Drielly; Tavares-Dias, Marcos
This study investigated the community of ectoparasites and endoparasites in Ageneiosus ucayalensis (Auchenipteridae) of a tributary from the Amazon River system, in Northern Brazil. Of 34 fish examined, 100% were parasitized by Ichthyophthirius multifiliis , Cosmetocleithrum bulbocirrus , Demidospermus sp., metacercariae of Genarchella genarchella , Clinostomum marginatum and Herpetodiplostomum sp., Procamallanus ( Spirocamallanus ) belenensis , Cucullanus ageneiosus and larvae of Contracaecum sp. digeneans C. marginatum and Herpetodiplostomum sp. were dominant parasite species, while I. multifiliis was the parasite with higher infection level. Such parasite species showed an aggregated dispersion, except P. ( S. ) belenensis , which showed a random dispersion. The Brillouin diversity (0.53 ± 0.29) was high, while evenness (0.28 ± 0.16) and species richness of parasites (3.7 ± 1.1) were low. The size of the hosts did not influence diversity, species richness and abundance of parasites. The ectoparasites were characterized by high prevalence and abundance, while endoparasites community presented low prevalence and abundance. The main factors responsible for structuring the parasite community in A. ucayalensis were mainly the behavior of this host and the availability of endoparasites infective stages in the environment. This was the first report of I. multifiliis , C. bulbocirus , Demidospermus sp., Contracaecum sp., C. marginatum , Herpetodiplostomum sp. and G. genarchella for A. ucayalensis . The presence of endohelminth larvae suggests that A. ucayalensis is part of the diet of other fish at the top of the food web in the Amazonian ecosystem studied.
Michelle Daniele Santos-Clapp
Full Text Available A total of 112 specimens of yellow peacock bass, Cichla kelberi, were collected in the Três Marias Reservoir, upper São Francisco River, State of Minas Gerais, between 2004 and 2005. Twelve species of parasites were collected: two species of ectoparasites (the monogenetic Gussevia sp. and the copepod Ergasilus sp.; and ten species of endoparasites, including four species of adult endoparasites: the cestodes Proteocephalus macrophallus and P. microscopicus, and the nematodes: Capillostrongyloides sentinosa and Rhabdochona acuminata, and six species of larval endoparasites: the metacercariae of Austrodiplostomum compactum and the nematodes: Hysterothylacium sp., Contracaecum sp. Type 1, Procamallanus (Spirocamallanus inopinatus, Spiroxys sp., and Goezia sp.. A significant positive correlation in the abundance of the two adult endoparasites species, P. macrophallus and P. microscopicus, may indicate that such associated species had the same ecological requirements. The positive correlations between the prevalence of the larval endoparasites, Hysterothylacium sp. and A. compactum, may indicate that there was opportunism for the two generalists species. Ergasilus sp., C. sentinosa, R. acuminata and Hysterothylacium sp. were recorded for the first time in C. kelberi, and this increased their list of known hosts. In addition, both proteocephalid species and C. sentinosa can extend their known biogeographical distribution to the Três Marias Reservoir.
Full Text Available This study assessed the helminth fauna of Astyanax fasciatus in two distinct sites of the Taquari River, São Paulo State, with 30 individuals sampled in a lotic site and 30 in a lentic site, recording the monogeneans: Cacatuocotyle paranaensis, Characithecium costaricensis, Diaphorocleidus kabatai, Jainus sp., Notozothecium sp. and Gyrodactylus sp., the digenean Antorchis lintoni and no-identified metacercariae; the nematode Procamallanus (Spirocamallanus inopinatus and no-identified larvae. The mean abundances of total monogeneans (U = 1053; p = 0.042 and C. costaricensis (U = 1107; p = 0.005 were higher in the lotic site. This difference may be due to the higher density of the host population in the lotic site, and the water transparence in lentic environments that prevents A. fasciatus to form shoals, precluding the exchange of parasites with direct cycle within a host population. This study is the first report of the helminth fauna of A. fasciatus in the Taquari River, with ten taxa recorded, and reports A. fasciatus as a new host for Notozothecium sp. and C. paranaensis.
A. G. Rybka
Full Text Available The paper confirms long-time opisthorhis invasion role as a risk factor of host immune system reconstitution as well as an important factor in holangiocarcinomas development. It was shown that opisthorhosis invasion primal stage induce host immune system reconstitution. Host immune B-cells system is activated by metacercaria antigens, while the same antigens inhibits T-cells activity. Opisthorhis metabolites stimulate proliferative mithogen-induced T-cells acti vity. Chronic opisthorchis invasion leads to immune system disbalance. It means: decrease of specific and non-speci fic natural killers activity, number of high proliferative activity T-lymphocytes and the shift of regulatory T-cells subset to suppressors prevalence. At the same time specific as well as non-specific T-suppressors functional ability is very low. It was shown T-cells helper-amplifier activation. Despite of circulating B-cells decrease the antibody produced cells number is spleen increases significantly at the same time with circulating immune complexes accumulation. Even 3–6 month after dehelmintisation the immune system disbalance decreases but lefts. In addition, chronic opisthorhis invasion leads to the proliferative processes activation in ductal epithelium, liver, lymph nodes and in other organs which leads to cancer proliferation. According to the results obtained the opisthorhis infected patients needs to be immunocorrected before as well as after dehelmintisation for holangiocancerogenesis profylaxis.
Ganzha, E V; Granovich, A I
Sporocycts of the "pygmaeus" microphallides (Microphallus piriformes) are localized in hepatopancreas and gonads of Littorina molluscs causing total parasitic castration. A histological study of penial glands in Littorina saxatilis and L. obtusata males infested with trematodes M. piriformes has been made. Copulatory organs of noninfested molluscs, molluscs after recent contamination (with not completely formed daughter sporocysts), and molluscs containing mature metacercariae inside daughter sporocysts were examined. Based on the data obtained, probable dynamics of the histological structure of infested glandular apparatus was established. It was shown, that the trematode infestation have an influence on the muscular and secretory parts of penial glands. The wall of the penial gland muscular capsule becomes more fine in infested L. saxatilis. On the contrary, this wall is vastly thicker in infested L. obtusata, as compared with noninfested individuals. Glandular cells of the molluscs' penial glands decreases the amount of granular secret in both species. In L. obtusata the number of secretory cells is shown to be reduced up to their total disappearance. The above pathological changes probably prevent normal function of penial glands.
Blazer, Vicki; Iwanowicz, Luke R.; Starliper, Clifford E.; Zaugg, Steven D.; Burkhardt, Mark R.; Barbash, P.; Hedrick, J.D.; Reeser, S.J.; Mullican, J.E.; Kelble, J.
Skin lesions and spring mortality events of smallmouth bass Micropterus dolomieu and selected other species were first noted in the South Branch of the Potomac River in 2002. Since that year morbidity and mortality have also been observed in the Shenandoah and Monocacy rivers. Despite much research, no single pathogen, parasite, or chemical cause for the lesions and mortality has been identified. Numerous parasites, most commonly trematode metacercariae and myxozoans; the bacterial pathogens Aeromonas hydrophila, Aeromonas salmonicida, and Flavobacterium columnare; and largemouth bass virus have all been observed. None have been consistently isolated or observed at all sites, however, nor has any consistent microscopic pathology of the lesions been observed. A variety of histological changes associated with exposure to environmental contaminants or stressors, including intersex (testicular oocytes), high numbers of macrophage aggregates, oxidative damage, gill lesions, and epidermal papillomas, were observed. The findings indicate that selected sensitive species may be stressed by multiple factors and constantly close to the threshold between a sustainable (healthy) and nonsustainable (unhealthy) condition. Fish health is often used as an indicator of aquatic ecosystem health, and these findings raise concerns about environmental degradation within the Potomac River drainage. Unfortunately, while much information has been gained from the studies conducted to date, due to the multiple state jurisdictions involved, competing interests, and other issues, there has been no coordinated approach to identifying and mitigating the stressors. This synthesis emphasizes the need for multiyear, interdisciplinary, integrative research to identify the underlying stressors and possible management actions to enhance ecosystem health.
Kaewpitoon, Natthawut; Kootanavanichpong, Nusorn; Kompor, Ponthip; Chavenkun, Wasugree; Kujapun, Jirawoot; Norkaew, Jun; Ponphimai, Sukanya; Matrakool, Likit; Tongtawee, Taweesak; Panpimanmas, Sukij; Rujirakul, Ratana; Padchasuwan, Natnapa; Pholsripradit, Poowadol; Eksanti, Thawatchai; Phatisena, Tanida; Loyd, Ryan A; Kaewpitoon, Soraya J
Opisthorchis viverrini is remains a public health problem in Thailand, particularly in the northeast and north regions which have the highest incidences of chonalgiocarcinoma (CCA). O. viverrini causes the disease opithorchiasis, and its has been classified as a group 1 biological carcinogen. Humans, dogs, and cats become infected with O. viverrini by ingesting raw or undercooked fish containing infective metacercariae. The first human cases of O. viverrini infection were reported in Thailand 100 years ago, and it's still a problem at the community level. Based on data for the year 2009, more than 6 million people were infected with O. viverrini. Associated medical care and loss of wages in Thailand costs about $120 million annually. This review highlights the current status of O. viverrini infection in communities of Thailand through active surveillance for the five years period from 2010 and 2015. A total of 17 community-based surveys were conducted, most in the northeast region. Some 7 surveys demonstrated a high prevalence over 20%, and the highest was 45.7%. Most commonly infection was found in age group of 35 years and older, males, and agricultural workers. Although, the national prevalence may be decreasing but the results show that the O. viverrini infection is still high in communities of the northeast region. Therefore, the focus in populations living in northeast Thailand should be screening of infection and changing their eating behavior.
Doanh, P N; Hien, H V; Nonaka, N; Horii, Y; Nawa, Y
Among about 50 nominal Paragonimus species, Paragonimus proliferus is rather a rare species, found only in Yunnan province, China, until our recent discovery of this species in Lai Chau province, northern Vietnam close to Yunnan, China. Here we add Quang Binh province, central Vietnam as a new endemic area of P. proliferus. Large excysted metacercariae found in mountainous crabs, Potamiscus tannanti, were morphologically identified as P. proliferus, which was confirmed further by molecular analyses. Second internal transcribed spacer (ITS2) sequences of the P. proliferus population in Quang Binh province were completely (100%) identical with those of P. proliferus populations in Lai Chau province, northern Vietnam and Yunnan province, China. However, cytochrome oxidase subunit 1 (CO1) gene sequences of Quang Binh population were significantly different (5.6%) from that of previously reported northern Vietnam and Yunnan, China populations. A phylogenetic tree revealed that all CO1 sequences of P. proliferus Quang Binh population formed a distinct group, which was clustered with northern Vietnam and Yunnan, China populations with the bootstrap value of 75%. This is the first record of the genetically variant population of P. proliferus, distribution of which is geographically remote from the previously reported endemic areas in the border between northern Vietnam and Yunnan, China, suggesting that P. proliferus may be much more widely distributed in the Indochina peninsula (or South-East Asia) than expected.
Borges, J N; Costa, V S; Mantovani, C; Barros, E; Santos, E G N; Mafra, C L; Santos, C P
Pygidiopsis macrostomum and Ascocotyle (Phagicola) pindoramensis (Digenea: Heterophyidae) parasitize guppies as intermediate hosts and, respectively, fish-eating mammals or birds as definitive hosts. Heterophyids have zoonotic potential, and molecular studies associated with morphological and ecological aspects have helped to clarify their taxonomy and phylogeny. Poecilia vivipara naturally parasitized by metacercariae of both species (100% prevalence) exhibit no external signs of parasitism. In this work, four new sequences of P. macrostomum (18S rDNA, 28S rDNA and ITS2 rDNA) and one new sequence of A. (P.) pindoramensis (mtDNA cox-1) are presented. Phylogeny reconstructions linked P. macrostomum to other heterophyids, but the separation of the Heterophyidae and Opisthorchiidae remains unclear. Additionally, we used indirect immunocytochemistry and the phalloidin-fluorescence techniques allied with confocal laser scanning microscopy to describe muscular and neuronal structures of P. macrostomum. A complex arrangement of muscular fibres is associated with the tegument, suckers, gut and reproductive system. Radial fibres around the ventral sucker are thick, branched and extend to the body wall. High-resolution confocal imaging revealed a typical digenean muscular arrangement and important heterophyid morphological traits. These data will support future control measures to reduce the parasitism in guppies reared in fish farming systems, especially for aquarium and experimental purposes. © 2016 John Wiley & Sons Ltd.
Endohelminth parasites of the freshwater fish Zoogoneticus purhepechus (Cyprinodontiformes: Goodeidae from two springs in the Lower Lerma River, Mexico Endohelmintos parásitos del pez dulceacuícola Zoogoneticus purhepechus (Cyprinodontiformes: Goodeidae en dos manantiales de la cuenca del río Lerma bajo, México
Full Text Available In order to establish the helminthological record of the viviparous fish species Zoogoneticus purhepechus, 72 individuals were collected from 2 localities, La Luz spring (n= 45 and Los Negritos spring (n= 27, both in the lower Lerma River, in Michoacán state, Mexico. Twelve helminth taxa were recovered, 5 adults (the digeneans Margotrema bravoae and Phyllodistomum sp., the cestode Bothriocephalus acheilognathi, the nematode Rhabdochona lichtenfelsi and the acanthocephalan Pomphorhynchus cf. bulbocolli, and 7 larvae (the metacercariae of Clinostomum complanatum, the cysticercoid of Cyclophyllidea, the nematodes Rhabdochona sp., Eustrongylides sp., Contracaecum sp. and Spiroxys sp., and the cysthacanth of Polymorphus brevis. Of these, R. lichtenfelsi was the most prevalent and abundant species at La Luz spring with 15.6% and 0.33 individuals per analyzed host. The remaining species were relatively more rare and infrequent. The helminth parasite community of Z. purhepechus at Los Negritos spring was remarkably poor and abundance was very low. The omnivorous feeding habits, the position of the host species in the food web, and the environmental characteristics of each locality are suggested as the main factors determining the helminth parasite communities in this freshwater fish.Se examinaron 72 individuos del pez vivíparo Zoogoneticus purhepechus para establecer el registro helmintológico de la especie. Los huéspedes se recolectaron de los manantiales La Luz (n= 45 y Los Negritos (n= 27, ubicados en la porción baja del río Lerma, en el estado de Michoacán, México. El registro helmintológico consta de 12 especies, incluyendo como adultos los digéneos Margotrema bravoae y Phyllodistomum sp., el céstodo Bothriocephalus acheilognathi, el nemátodo Rhabdochona lichtenfelsi, y el acantocéfalo Pomphorhynchus cf. bulbocolli. Además, como estadios larvarios, se encontraron las metacercarias de Clinostomum complanatum, el cisticercoide de
Coproantígenos de Fasciola hepatica de posible utilidad en el diagnóstico de la fascioliasis Identification and isolation of coproantigens of Fasciola hepatica that are potentially useful in the diagnosis of fascioliasis
Ana M. Espino
Full Text Available En el presente estudio se describen cuáles son los antígenos de Fasciola hepatica que están presentes en las heces de pacientes con fascioliasis crónica y de ratas infectadas experimentalmente con metacercarias de F. hepatica. Mediante el empleo de la técnica de inmunoelectrotransferencia (Western blot con un suero hiperinmune obtenido frente a antígenos de excreción/secreción de adultos de F. hepatica, se pudo demostrar que en las heces de los pacientes se encuentran antígenos de posible interés para el diagnóstico con masas moleculares de 14, 19, 20, 23, 25, 32, 46, 51 y 62 kilodaltons (kDa. Además, se comprobó que algunos de estos péptidos (los de 14, 20, 23 y 51 kDa son reconocidos también por la mayoría de los sueros de pacientes crónicos. Después de purificar por cromatografía de afinidad los antígenos presentes en heces de ratas con 6 a 12 semanas de infección, empleando para ello el anticuerpo monoclonal ES78 acoplado a Sepharosa 4B-CNBr, se identificaron seis polipéptidos de 11, 14, 26, 32, 47 y 51 kDa; otros tres polipéptidos, de 17, 24 y 66 kDa, solo se pudieron identificar en heces de ratas con 10 a 12 semanas de infección. Nuestros resultados sugieren que estos polipéptidos pueden ser antígenos comunes a ambos estadios parasitarios y constituir importantes marcadores de la fascioliasis aguda y crónica, en particular los de 14, 24, 26 y 51 kDa, puesto que reaccionaron con el inmunosuero, los sueros humanos y el anticuerpo monoclonal ES78.This study describes which antigens of Fasciola hepatica are present in the feces of patients with chronic fascioliasis and in the feces of rats infected experimentally with F. hepatica metacercariae. Using a Western blot assay technique with hyperimmune serum obtained from excretory-secretory antigens of adult F. hepatica, we found in the patients' feces antigens of possible diagnostic interest, with molecular weights of 14, 19, 20, 23, 25, 32, 46, 51, and 62 kilodaltons
Dung, Bui Thi; Madsen, Henry; The, Dang Tat
Fish-borne zoonotic trematodes, such as Clonorchis sinensis, heterophyids and others, constitute a public health concern in parts of northern Vietnam and infections with these trematodes are often thought to be linked to fish culture. One common fish culture system is the integrated fish-livestock (VAC) ponds where individual households have 1 or more ponds. Fish fry, mainly of various carp species, produced in hatcheries, not necessarily local, are introduced into nursery ponds and after approximately 6 weeks, juvenile fishes are transferred to household ponds, referred to as grow-out ponds. Grow-out ponds are usually fertilized with organic debris, including animal excreta, to stimulate algal growth and subsequently fish growth. This paper describes the distribution of freshwater snails and occurrence of trematode infections in these in VAC ponds and associated habitats as part of a major study on risk factors of FZT infections in cultured fish in two communes, Nghia Lac and Nghia Phu, Nghia Hung District, Nam Dinh Province. The area is under intense rice cultivation with an extensive canal network supplying fields and also household VAC ponds. A total of 16 snail species was found and four were widely distributed i.e. Angulyagra polyzonata, Melanoides tuberculata, Bithynia fuchsiana and Pomacea insularum. Snail diversity and counts were higher in nursery ponds than in grow-out ponds. Species of the families Thiaridae and Viviparidae were more abundant than other species in VAC ponds while species of the Bithyniidae, Stenothyridae and Planorbidae dominated in rice fields and small canals. Trematode infections were found in eight snail species and among these M. tuberculata had the highest overall prevalence of infection (13.28%). No trematode infections were found in species of the Viviparidae and Ampullaridae except for metacercariae. Parapleurolophocercous and pleurolophocercous cercariae constituted the most common type of cercariae recovered, contributing 40
Hamann, M I; Kehr, A I; González, C E
The main goals of this study were to compare the richness of parasitic trematodes in amphibians with diverse habits (terrestrial, fossorial, semi-aquatic and arboreal), and to evaluate whether the composition of the trematode community is determined by ecological relationships. Specimens were collected between April 2001 and December 2006 from a common area (30 ha) in Corrientes Province, Argentina. Trematodes of amphibians in this area comprised a total of 19 species, and were dominated by common species. Larval trematodes presented highest species richness, with the metacercaria of Bursotrema tetracotyloides being dominant in the majority (7/9, 78%) of the parasite communities. Adults of the trematode Catadiscus inopinatus were dominant in the majority (6/9, 67%) of amphibians. The amphibians Leptodactylus latinasus, Leptodactylus bufonius and Scinax nasicus presented a high diversity of trematodes, whereas Leptodactylus chaquensis had the lowest diversity even though it presented with the highest species richness. The patterns of similarity among amphibian species showed groups linking with their habitats. Leptodactilid amphibians, with a generalist diet and an active foraging strategy showed highest infection rates with adult trematodes. The mean richness of trematode species related to host's habitat preferences was higher in semi-aquatic amphibians. Results suggest that semi-aquatic amphibians, present in both aquatic and terrestrial environments, present a greater diversity of parasites as they have a higher rate of exposure to a wider range of prey species and, hence, to diverse infective states. The trematode composition is related to the diets and mobility of the host, and habitat.
Full Text Available Introduction & Objective: Fascioliasis is a zoonotic parasitic disease, caused by the liver fluke, Fasciola spp.. Human is occasional host when ingesting the metacercaria by eating contaminated aquatic vegetable. In the two past decades, human fasciolasis was emerging as a problem of public health in the Guilan province; in Anzali city. Triclabendazole is a novel anti-helmenthic that during recent years has been used for fascioliasis treatment in this region. The aim of the present work is to study alteration in haematological and liver function indices during human infection with Fasciola spp. pre and post treatment with triclabendazoleMaterials & Methods: The present work is a longitudinal clinical trail. In this regard, fifty confirmed fasciolasis patients, were chosen for parasitological, hematological and biochemical examinations pre-therapy as well as 1 and 6 months post-therapy. Formalin-ether and modified Telemann methods were used for stool examination. For Fasciola antibody detection ELISA technique was employed. Hematological and biochemical tests were performed by standard methods. Results: Results indicated that, triclabendazole efficacy was 74% after usage as one dose of 20mg/kg and reached to 88% after repeating in the next month. Before triclabebdazole therapy the Hb and HCT of the patients were slightly found lower than normal ranges, meanwhile the ESR and eosinophil percentages were higher. However following receiving the drug, in the cured individuals, the indices returned to the normal ranges but in the non-cured individuals were not shifted to the normal. On the other hand liver function indices of the patients mostly were at normal ranges before and following drug therapy.Conclusion: In conclusion haematological indices could be valuable indicator for successful therapy of patients treated with triclabendazole.
Villeneuve, Daniel L; Curtis, Lawrence R; Jenkins, Jeffrey J; Warner, Kara E; Tilton, Fred; Kent, Michael L; Watral, Virginia G; Cunningham, Michael E; Markle, Douglas F; Sethajintanin, Doolalai; Krissanakriangkrai, Oraphin; Johnson, Eugene R; Grove, Robert; Anderson, Kim A
The Willamette River, one of 14 American Heritage Rivers, flows through the most densely populated and agriculturally productive region of Oregon. Previous biological monitoring of the Willamette River detected elevated frequencies of skeletal deformities in fish from certain areas of the lower (Newberg pool [NP], rivermile [RM] 26 - 55) and middle (Wheatland Ferry [WF], RM 72 - 74) river, relative to those in the upper river (Corvallis [CV], RM 125-138). The objective of this study was to determine the likely cause of these skeletal deformities. In 2002 and 2003, deformity loads in Willamette River fishes were 2-3 times greater at the NP and WF locations than at the CV location. There were some differences in water quality parameters between the NP and CV sites, but they did not readily explain the difference in deformity loads. Concentrations of bioavailable metals were below detection limits (0.6 - 1 microg/ L). Concentrations of bioavailable polychlorinated biphenyls (PCBs) and chlorinated pesticides were generally below 0.25 ng/L. Concentrations of bioavailable polycyclic aromatic hydrocarbons were generally less than 5 ng/L. Concentrations of most persistent organic pollutants were below detection limits in ovary/oocyte tissue samples and sediments, and those that were detected were not significantly different among sites. Bioassay of Willamette River water extracts provided no evidence that unidentified compounds or the complex mixture of compounds present in the extracts could induce skeletal deformities in cyprinid fish. However, metacercariae of a digenean trematode were directly associated with a large percentage of deformities detected in two Willamette River fishes, and similar deformities were reproduced in laboratoryfathead minnows exposed to cercariae extracted from Willamette River snails. Thus, the weight of evidence suggests that parasitic infection, not chemical contaminants, was the primary cause of skeletal deformities observed in Willamette
Juan Carlos Casanova
Full Text Available Prospections on helminth fauna of rodents were carried out in the Cedrino river (Eastern of Sardinia island. Twelve Rattus rattus (L., 1758 and eight Mus domesticus (Schwarz & Schwarz, 1943 were captured by Sherman traps. Parasitological study revealed in R. rattus the presence of an intestinal helminth belonging to the genus Maritrema (family Microphallidae. The transmission of this Digenetic Trematode among vertebrates involves the participation of aquatic invertebrate organisms, molluscs and crustacea, acting as intermediate hosts. Dissection of 117 individuals of the Amphipod Crustacea Gammarus italicus, collected in the same biotope, allowed the detection of encysted metacercariae. These larvae constitute the infesting stage of the parasite for vertebrates, acting these as definitive hosts ingesting parasitized crustacea as preys. Adults of Maritrema sp. were also obtained experimentally in the laboratory mice (Mus domesticus CD1 strain. These were infested by inoculating them, using gastric probe, metacercarial cysts isolated from G. italicus. Post-infection mice dissection was performed at different intervals of time getting adults in various maturity stages. Experimental facts confirm that Maritrema sp., as it occurs in other trematodes, do not present strict specificity to the definitive host, being able to develop as well in Mus. In nature, the presence or absence of this digenean in mammals species will be dependent of host ethological factors, mainly related to feeding habitats. This study was partially supported by the ?Comissionat per Recerca i Universitats de la Generalitat de Catalunya? 2001SGR00088.
Pérez, José; Ortega, Joaquín; Bravo, Ana; Díez-Baños, Pablo; Morrondo, Patrocinio; Moreno, Teodoro; Martínez-Moreno, Alvaro
The phenotype of the hepatic inflammatory infiltrate and hepatic lymph nodes (HLN) was analysed in lambs primarily and challenge infected with Fasciola hepatica. Group 1 was primarily and challenge infected with two doses of 200 metacercariae (mc) each and was non-treated. Trickle infection was administered to five groups: group 2 was challenge infected and non-treated; group 3 was primarily infected and non-treated; group 4 was primarily infected and treated with triclabendazole (TCBZ) at 12 weeks postinfection (wpi); group 5 was treated at 4 wpi and challenge infected and group 6 was treated at 12 wpi and challenge infected. An uninfected group was used as the control. The distribution of T cell subpopulations (CD3, CD4 and CD8), and B cells (CD79alpha, IgM, IgG) was analysed. The hepatic inflammatory infiltrate was represented mainly by CD3 and CD4 T cells, and B cells (CD79alpha, IgG). These infiltrates were more severe (P < 0.05) in primarily (group 3) or challenge (groups 2, 5 and 6) trickle infected lambs than in the group single challenge infected (group 1). Cellular changes in HLN consisted in an increase of CD4 over CD8 T cells and an increase of B cells and IgG+ plasma cells, and they were more severe in primarily and challenge trickle infected groups than in the group infected with two larger doses of mc, although significant differences were not found with respect to all challenge trickle infected groups. The strong local cellular and humoral immune responses did not protect against subsequent infections, neither in non-treated lambs (group 2) nor in lambs treated with TCBZ at 4 wpi (group 5) or 12 wpi (group 6).
Zhang, Fu-Kai; Zhang, Xiao-Xuan; Elsheikha, Hany M; He, Jun-Jun; Sheng, Zhao-An; Zheng, Wen-Bin; Ma, Jian-Gang; Huang, Wei-Yi; Guo, Ai-Jiang; Zhu, Xing-Quan
Fasciola gigantica, the tropical liver fluke, infects buffaloes in Asian and African countries and causes significant economic losses and poses public health threat in these countries. However, little is known of the transcriptional response of buffaloes to infection with F. gigantica. The objective of the present study was to perform the first transcriptomic analysis of buffalo liver infected by F. gigantica. Understanding the mechanisms that underpin F. gigantica infection in buffaloes will contribute to our ability to control this parasite. We challenged buffaloes with 500 viable F. gigantica metacercariae and collected liver samples through a time course at 3, 42 and 70 days post-infection (dpi). Then, we performed gene expression analysis on liver samples using RNA sequencing (RNA-Seq) Illumina technology and confirmed the RNA-Seq data by quantitative RT-PCR analysis. Totals of 496, 880 and 441 differentially expressed transcripts were identified in the infected livers at 3, 42 and 70 dpi, respectively. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis revealed that transcriptional changes in the liver of infected buffaloes evolve over the course of infection. The predominant response of buffaloes to infection was mediated by certain pathways, such as MHC antigen processing and presentation, Toll-like receptor 4 (TLR4), transforming growth factor beta (TGF-β), and the cytochrome P450. Hepatic drug metabolizing enzymes and bile secretion were also affected. Fasciola gigantica can induce statistically significant and biologically plausible differences in the hepatic gene expression of infected buffaloes. These findings provide new insights into the response of buffaloes to F. gigantica over the course of infection, which may be useful in determining pathways that can modulate host-parasite interaction and thus potentially important for clearance of the parasite.
Escamilla, A; Bautista, M J; Zafra, R; Pacheco, I L; Ruiz, M T; Martínez-Cruz, S; Méndez, A; Martínez-Moreno, A; Molina-Hernández, V; Pérez, J
The aim of the present work was to evaluate the number of apoptotic eosinophils in the livers of sheep experimentally infected with Fasciola hepatica during the migratory and biliary stages of infection. Four groups (n=5) of sheep were used; groups 1-3 were orally infected with 200 metacercariae (mc) and sacrificed at 8 and 28 days post-infection (dpi), and 17 weeks post-infection (wpi), respectively. Group 4 was used as an uninfected control. Apoptosis was detected using immunohistochemistry with a polyclonal antibody against anti-active caspase-3, and transmission electron microscopy (TEM). Eosinophils were identified using the Hansel stain in serial sections for caspase-3, and by ultrastructural features using TEM. At 8 and 28 dpi, numerous caspase-3(+) eosinophils were mainly found at the periphery of acute hepatic necrotic foci. The percentage of caspase -3(+) apoptotic eosinophils in the periphery of necrotic foci was high (46.1-53.9) at 8 and 28 dpi, respectively, and decreased in granulomas found at 28 dpi (6%). Transmission electron microscopy confirmed the presence of apoptotic eosinophils in hepatic lesions at 8 and 28 dpi. At 17 wpi, apoptotic eosinophils were detected in the infiltrate surrounding some enlarged bile ducts containing adult flukes. This is the first report of apoptosis induced by F. hepatica in sheep and the first study reporting apoptosis in eosinophils in hepatic inflammatory infiltrates in vivo. The high number of apoptotic eosinophils in acute necrotic tracts during the migratory and biliary stages of infection suggests that eosinophil apoptosis may play a role in F. hepatica survival during different stages of infection. Copyright © 2015 Elsevier B.V. All rights reserved.
Choudhury, A.; Hoffnagle, T.L.; Cole, Rebecca A.
A 2-yr, seasonal, parasitological study of 1,435 fish, belonging to 4 species of native fishes and 7 species of nonnative fishes from the lower Little Colorado River (LCR) and tributary creeks, Grand Canyon, Arizona, yielded 17 species of parasites. These comprised 1 myxozoan (Henneguya exilis), 2 copepods (Ergasilus arthrosis and Lernaea cyprinacea), 1 acarine (Oribatida gen. sp.), 1 piscicolid leech (Myzobdella lugubris), 4 monogeneans (Gyrodactylus hoffmani, Gyrodactylus sp., Dactylogyrus extensus, and Ligictaluridus floridanus), 4 nematodes (Contracaecum sp., Eustrongylides sp., Rhabdochona sp., and Truttaedacnitis truttae), 3 cestodes (Bothriocephalus acheilognathi, Corallobothrium fimbriatum, and Megathylacoides giganteum), and 2 trematodes (Ornithodiplostomum sp. and Posthodiplostomum sp.). Rhabdochona sp. was the only adult parasite native to the LCR. Infection intensities of Ornithodiplostomum sp. and B. acheilognathi were positively correlated with length of the humpback chub Gila cypha. Adult helminths showed a high degree of host specificity, except B. acheilognathi, which was recovered from all fish species examined but was most abundant in cyprinids. Abundance of B. acheilognathi in the humpback chub was highest in the fall and lowest in the summer in both reaches of the LCR. There was no major taxonomic difference in parasite assemblages between the 2 different reaches of the river (LC1 and LC2). Parasite community diversity was very similar in humpback chub, regardless of sampling site or time. The parasite fauna of the LCR is numerically dominated by B. acheilognathi and metacercariae of Ornithodiplostomum sp. The richest and most diverse component community occurred in a nonnative species, the channel catfish Ictalurus punctatus, but infracommunity species richness was highest in a native host, humpback chub.
Full Text Available Paragonimus rudis was found in the lungs of a giant otter Lutra (pteronura brasiliensis by Natterer in 1828, who dissected the animal in the former capital Mato Grosso (=Vila Bela, Brazil. The flukes were described by Diesing in 1850, and redescribed by Braun in 1901. Both descriptions do not allow to identify the species. Therefore, P. rudis must be regarded a "nomen nudum". Because its rediscovery is desirable with regard to historical reasons and nomenclatoric questions, a field study was performed in Mato Grosso in 1980. Of 354 freshwater crabs from 24 localities collected and examined for parasitic infections, about 25% were found to be infected with 7kinds of trematode larvae, which differed distincly from Paragonimus-metacercariae. The question, whether P. rudis or other lung fluke species do not seem to occur or cannot be found any longer in the area investigated by us, is discussed.Em 1828, Natterer encontrou o Paragonimus rudis nos pulmões de uma lontra gigante Lutra (Pteronura brasiliensis, Vila Bela (Mato Grosso Brasil. Os vermes foram descritos em 1850 por Diesing e novamente em 1909 por Braun. Ambas descrições nãopermitem a identificação das espécies e portanto P. rudis pode ser tido como "nomennudum". Foi feita uma tentativa de se encontrar outros exemplares examinando 354 caranguejos de 24 localidades. Cerca de 25% deles estavam infectados com sete tipos de larvas de trematódeos que diferem das metacercárias de Paragonimus. Assim permanece sem resposta a pergunta: O que é o Paragonimus rudi?.
Arias, Sandra Milena; Salazar, Lina Marcela; Casas, Eudoro; Henao, Alexandra; Velásquez, Luz Elena
Previous research has demonstrated the presence of Paragonimus spp. in crabs of three municipalities located in the Medellín river basin. To design a plan for the control and prevention of paragonimosis, parasite presence must be demonstrated in the upper area of the river, and an awareness campaign of its risks must involve the participation of the educational community in focusing attention on the local aquatic resources. The presence of Paragonimus spp. metarcercariae was established in freshwater crabs from La Miel and La Clara streams, and an awareness campaign created public awareness to management of the aquatic ecosystems for risk reduction. Nine crabs were collected from streams of La Miel and La Clara villages in 2007 and 2008. The crabs were indentified, sacrificed in the laboratory and examined for presence of Paragonimus spp. metacercariae. Five workshops were conducted with students of educational institutions of the two villages to improve the student perception of paragonimosis risks associated with the local streams. The crabs were identified as Hypolobocera bouvieri. One crab was found with Paragonimus spp. in La Miel. A mollusk, Aroapyrgus colombiensis, another host of the parasite was found in La Clara. That aquatic ecosystems are a great attraction for the local was well established. Foci of Paragonimus spp. flatworms are present in the upper area of Medellín river which includes Caldas. However, local school children were uninformed about the risk of paragonimosis associated with the local aquatic ecosystem. Therefore this province must be included in programs for prevention and control of paragonimosis. The prevalence of this disease recommends educational programs that promote awareness of local aquatic ecosystems and their parasitic inhabitants.
Jiang, Hongye; Chen, Tingjin; Sun, Hengchang; Tang, Zeli; Yu, Jinyun; Lin, Zhipeng; Ren, Pengli; Zhou, Xinyi; Huang, Yan; Li, Xuerong; Yu, Xinbing
Clonorchiasis, caused by the consumption of raw or undercooked freshwater fish containing infective metacercariae of Clonorchis sinensisis (C.sinensis), remains a common public health problem. New effective prevention strategies are still urgent to control this food-borne infectious disease. The previous studies suggested Bacillus subtilis (B. subtilis) spores was an ideal vaccines delivery system, and the C.sinensis enolase (CsENO) was a potential vaccine candidate against clonorchiasis. In the current study, we detected CsENO-specific IgM levels by ELISA in sera, intestinal mucus and skin mucus in grass carps (Ctenopharyngodon idella) through oral administration with B. subtilis spores surface expressing CsENO. In addition, immune-related genes expression was also measured by qRT-PCR. Grass carps orally treated with B. subtilis spores or normal forages were used as controls. The results of ELISA manifested that specific IgM levels of grass carps in CsENO group in sera, intestine mucus and skin mucus almost significantly increased from week 4 post the first oral administration when compared to the two control groups. The levels of specific IgM reached its peak in intestine mucus firstly, then in sera, and last in skin mucus. qRT-PCR results showed that 5 immune-related genes expression had different degree of rising trend in CsENO group when compared to the two control groups. Our study demonstrated that orally administrated with B. subtilis spores expressing CsENO induced innate and adaptive immunity, systemic and local mucosal immunity, and humoral and cellular immunity. Our work may pave the way to clarify the exact mechanisms of protective efficacy elicited by B. subtilis spores expressing CsENO and provide new ideas for vaccine development against C. sinensis infection. Copyright © 2016 Elsevier Ltd. All rights reserved.
Huang, Yalan; Huang, Dana; Geng, Yijie; Fang, Shisong; Yang, Fan; Wu, Chunli; Zhang, Hailong; Wang, Miao; Zhang, Ran; Wang, Xin; Wu, Shuang; Cao, Jianping; Zhang, Renli
Clonorchis sinensis is an important foodborne zoonosis worldwide and prevalent in China for more than 2000 years. According to the experience of controlling Schistosoma japonica, China started to establish the integrated control strategy for C. sinensis in endemic areas. Lou village, the largest village in Shenzhen city in South China was taken as a pilot site. This longitudinal study assessed the infection status of C. sinensis among people and intermediate hosts from 2006 to 2014 in Lou village. After a continuous intervention with the integrated control strategy, the prevalence of C. sinensis decreased significantly to 2.01% in 2014. The infection intensity also reduced significantly with eggs per gram varying from 45.6 ± 3.4 in 2010 to 21.7 ± 1.6 in 2012. There is also a statistically significant decrease of the prevalence of C. sinensis metacercariae in fish hosts from 16.51% in 2008 before the intervention to 5.33% in 2014. All the old-styled toilets were replaced by sanitary ones with a harmless processing design in 2014. No viable parasite eggs were detected in stool samples from the reconstructed toilets. Health education played an important role in changing the eating habits among the local residents, with a significant decrease in the prevalence of eating raw fish from 91.99% in 2008 to 59.87% in 2014. The evaluation suggested that the integrated strategy we have performed in Lou village is effective in controlling the C. sinensis infection and maintaining the infection rate at a lower level, which can be promoted in other endemic areas.
Hua, Hui; Du, Ying; Ma, Rui; Zhang, Bei-Bei; Yu, Qian; Li, Bo; Xu, Jiang-Tao; Li, Xiang-Yang; Tang, Ren-Xian; Yan, Chao; Zheng, Kui-Yang
The roles of TLR4 in mediation of innate immune response and in regulation of adaptive immune responses triggered by Clonorchis sinensis remain unknown. In the present study, splenocytes derived from C3H/HeN (TLR4 wild ) and C3H/Hej mice (TLR4 mut ) that were infected with 45 metacercariae of C. sinensis were harvested, then stimulated by C. sinensis excretory/secretory products (ESP) or medium (control) for 48 h, respectively. Meanwhile, bone marrow-derived dendritic cells (BMDCs) from normal C3H/HeN and C3H/Hej mice were prepared and stimulated with medium, ESP, LPS, or ESP+LPS for 24 h, respectively. The supernatants were collected, and the concentrations of type 1 and type 2 relative cytokines were determined by ELISA. The maturation of BMDCs indicated by surface markers of CD80, CD86, and MHC II was evaluated by flow cytometry. The results showed that the levels of IFN-γ, IL-6, TNF-α, and IL-10 in the splenocytes from C. sinensis-infected TLR4 mut mice were significantly lower than those from TLR4 wild mice when they were further exposed to ESP. For BMDCs, the productions of the cytokines IL-12p70 and IL-10, but not IL-4, in the BMDCs from TLR4 mutation mice were predominantly decreased compared with those from TLR4 wild mice when the BMDCs were co-stimulated by ESP combined with LPS. Flow cytometry analysis showed that ESP could significantly decrease the high levels of CD80, CD86, and MHC II which were elevated by LPS. In conclusion, these data suggest that TLR4 may play a regulatory role in type 1 immune responses during C. sinensis infection.
Tokuhiro, Shinji; Nagataki, Mitsuru; Jarilla, Blanca R.; Nomura, Haruka; Kim, Tae Im; Hong, Sung-Jong; Agatsuma, Takeshi
Background Adult Clonorchis sinensis lives in the bile duct and causes endemic clonorchiasis in East Asian countries. Phosphagen kinases (PK) constitute a highly conserved family of enzymes, which play a role in ATP buffering in cells, and are potential targets for chemotherapeutic agents, since variants of PK are found only in invertebrate animals, including helminthic parasites. This work is conducted to characterize a PK from C. sinensis and to address further investigation for future drug development. Methology/Principal findings A cDNA clone encoding a putative polypeptide of 717 amino acids was retrieved from a C. sinensis transcriptome. This polypeptide was homologous to taurocyamine kinase (TK) of the invertebrate animals and consisted of two contiguous domains. C. sinensis TK (CsTK) gene was reported and found consist of 13 exons intercalated with 12 introns. This suggested an evolutionary pathway originating from an arginine kinase gene group, and distinguished annelid TK from the general CK phylogenetic group. CsTK was found not to have a homologous counterpart in sequences analysis of its mammalian hosts from public databases. Individual domains of CsTK, as well as the whole two-domain enzyme, showed enzymatic activity and specificity toward taurocyamine substrate. Of the CsTK residues, R58, I60 and Y84 of domain 1, and H60, I63 and Y87 of domain 2 were found to participate in binding taurocyamine. CsTK expression was distributed in locomotive and reproductive organs of adult C. sinensis. Developmentally, CsTK was stably expressed in both the adult and metacercariae stages. Recombinant CsTK protein was found to have low sensitivity and specificity toward C. sinensis and platyhelminth-infected human sera on ELISA. Conclusion CsTK is a promising anti-C. sinensis drug target since the enzyme is found only in the C. sinensis and has a substrate specificity for taurocyamine, which is different from its mammalian counterpart, creatine. PMID:24278491
Full Text Available Because shifts in host–parasite relationships can alter host populations, attention should be given to the parasites that introduced species take with them or acquire in their introduced range. The Cuban treefrog, Osteopilus septentrionalis, is a successful invasive species in Florida with its parasites in the native range being well-documented, but there is a void in the literature regarding what parasites were lost or introduced in its expansion. We necropsied 330 O. septentrionalis from Tampa, FL and compared their macroparasites to those of O. septentrionalis in their native range and to the parasites of anurans native to the Tampa, FL area to determine the species O. septentrionalis likely introduced or acquired in Florida. At least nine parasite species (Aplectana sp., Oswaldocruzia lenteixeirai, Cylindrotaenia americana, Physaloptera sp., Rhabdias sp., Centrorhynchus sp., unidentified trematode metacercariae, unidentified larval acuariids, and unidentified pentastomids were isolated. We found no differences in parasite communities of adult male and female frogs, which averaged 19.36 parasite individuals and 1.39 parasite species per adult frog, and had an overall prevalence of 77.52%. Acuariid larvae were likely acquired by O. septentrionalis in FL because they are not found in their native range. O. lenteixeirai was likely introduced because it is commonly reported in O. septentrionalis' native range but has never been reported in FL-native anurans. Aplectana sp. is also likely introduced because it has been reported in several anurans in Cuba but only reported once in Florida. O. septentrionalis tended to harbor fewer of its native parasites in the introduced range, which is consistent with the enemy release hypothesis and potentially creates an immunological advantage for this invasive host. Because native populations can be threatened by introduced parasites, there is a need to further explore the frequency and rate at which
Full Text Available Previous studies showed that CD4+T cells responses might be involved in the process of biliary fibrosis. However, the underlying mechanism resulting in biliary fibrosis caused by Clonorchis sinensis remains not yet fully elucidated. The objectives of the present study were to investigate the different profiles of hepatic CD4+T cell subsets (Th1, Th2, Th17 and Treg cells and their possible roles in the biliary fibrosis of different strains of mice (C57BL/6, BALB/c and FVB mice induced by C. sinensis infection. C57BL/6, BALB/c and FVB mice were orally gavaged with 45 metacercariae. All mice were sacrificed on 28 days post infection in deep anesthesia conditions. The leukocytes in the liver were separated to examine CD4+T cell subsets by flow cytometry and the left lobe of liver was used to observe pathological changes, collagen depositions and the concentrations of hydroxyproline. The most serious cystic and fibrotic changes appeared in FVB infected mice indicated by gross observation, Masson's trichrome staining and hydroxyproline content detection. In contrast to C57BL/6 infected mice, diffuse nodules and more intensive fibrosis were observed in the BALB/c infected mice. No differences of the hepatic Th1 subset and Th17 subset were found among the three strains, but the hepatic Th2 and Treg cells and their relative cytokines were dramatically increased in the BALB/c and FVB infected groups compared with the C57BL/6 infected group (P<0.01. Importantly, increased Th2 subset and Treg subset all positively correlated with hydroxyproline contents (P<0.01. This result for the first time implied that the increased hepatic Th2 and Treg cell subsets were likely to play potential roles in the formation of biliary fibrosis in C. sinensis-infected mice.
Kudlai, Olena; Cribb, Thomas H; Cutmore, Scott C
A new species of microphallid, Longiductotrema tethepae n. sp., is reported from a muraenid eel Gymnothorax pseudothyrsoideus (Bleeker) on the northern Great Barrier Reef. The new species is described based on adults from Gy. pseudothyrsoideus and metacercariae from a grapsid crab, Grapsus albolineatus Latreille in Milbert, collected from off Lizard Island, Queensland, Australia. The new species is assigned to Longiductotrema Deblock & Heard, 1969 based on morphological characters (presence of a cirrus-sac; a long metraterm, intensively ensheathed by gland-cells; an entirely postcaecal uterus; vitellarium composed of two lateral clusters each of about ten follicles, situated in the testicular and post-testicular areas). Longiductotrema tethepae n. sp. is the third species assigned to the genus, differing from its congeners in having a distinctly larger body dimensions, a smaller pharynx in relation to oral sucker, the anterior limits of the vitelline fields at the level of the testes (vs at the level of the ovary) and in its parasitism of a muraenid fish (vs birds). Additionally, the new species differs from L. floridense Deblock & Heard, 1969 in having a shorter metraterm and from L. scandolensis Deblock & Bartoli, 1988 in having a less elongate forebody in relation to body length, shorter caeca and prepharynx, and slightly larger eggs. Phylogenetic analyses, based on partial 28S rRNA gene sequences, showed that the present species is sister to all other microphallids for which sequence data are available. This is the fourth report of a microphallid from a marine eel, the first from the Muraenidae Rafinesque and the first from a marine fish in the Indo-west Pacific. A summary of all species of the Microphallidae parasitising fish is provided.
Background Co-infections of multiple parasite species in hosts may lead to interspecific associations and subsequently shape the structure of a parasite community. However, few studies have focused on these associations in highly abundant parasite species or, in particular, investigated how the associations develop with time in hosts exposed to co-infecting parasite species for the first time. We investigated metacercarial establishment and interspecific associations in the trematodes Ichthyocotylurus variegatus and I. pileatus co-infecting three age cohorts of young perch (Perca fluviatilis). Results We found that the timing of transmission of the two Ichthyocotylurus species was very similar, but they showed differences in metacercarial development essentially so that the metacercariae of I. pileatus became encapsulated faster. Correlations between the abundances of the species were significantly positive after the first summer of host life and also within the main site of infection, the swim bladder. High or low abundances of both parasite species were also more frequent in the same host individuals than expected by chance, independently of host age or size. However, the highest abundances of the species were nevertheless observed in different host individuals and this pattern was consistent in all age cohorts. Conclusions The results suggest similar temporal patterns of transmission, non-random establishment, and facilitative rather than competitive associations between the parasite species independently of the age of the infracommunities. However, we suggest that spatial differences in exposure are most likely responsible for the segregation of the parasite species observed in the few most heavily infected hosts. Regardless of the underlying mechanism, the result suggests that between-species associations should be interpreted with caution along with detailed examination of the parasite distribution among host individuals. PMID:21599910
Boehs, Guisla; Villalba, Antonio; Ceuta, Liliane Oliveira; Luz, Joaldo Rocha
This paper reports the parasites found in three commercially exploited bivalve molluscs (Mytella guyanensis, Anomalocardia brasiliana and Iphigenia brasiliana) of an estuarine region of Ilhéus, south of Bahia, Brazil (14 degrees 48'23''S; 39 degrees 02'47''W). Samples of 20 individuals of each species were collected fortnightly from August 2005 to August 2006. A total of 1480 individuals was collected and processed by standard histologic techniques; the histologic sections were stained with Harris haematoxylin and eosin and examined with light microscope. The water temperature in the study area varied from 24 to 30.5 degrees C and the salinity from 0 to 23ppt. Remarkable differences were found in the parasitic community between the three mollusc species involved in the study, which occupied different habitats in the estuarine region of the Cachoeira river. The following parasites were found: intracellular rickettsia-like colonies in digestive epithelia; intracellular gregarine Nematopsis sp. in gills, mantle, gonad, digestive gland and foot muscle; sporocysts of a Bucephalidae trematode in gonads, mantle, gills, digestive gland and foot; unidentified digenetic metacercariae in digestive gland and gonad; metacestodes of Tylocephalum sp. in connective tissue in the digestive gland and in gonad; and an unidentified metazoan in mantle and intestinal lumen. No significant temporal variation in the prevalence of any parasite was detected, which could be due to the narrow temperature range of the region and the absence of patterns of salinity and rainfall variation through the year. The infestation by sporocyst was the only pathological threat detected for the studied populations because of its potential for host castration. The low infection intensity and/or prevalence of the other parasites and the lack of obvious lesions suggest that there is no other serious pathological risk for the studied mollusc populations.
Uhrig, Emily J; Spagnoli, Sean T; Tkach, Vasyl V; Kent, Michael L; Mason, Robert T
Trematodes of the genus Alaria develop into an arrested stage, known as mesocercariae, within their amphibian second intermediate host. The mesocercariae are frequently transmitted to a non-obligate paratenic host before reaching a definitive host where further development and reproduction can occur. Snakes are common paratenic hosts for Alaria spp. with the mesocercariae often aggregating in the host's tail. In the current study, we used morphological examination and molecular analyses based on partial sequences of nuclear large ribosomal subunit gene and mitochondrial cytochrome C oxidase subunit 1 gene to identify larvae in the tails of red-sided garter snakes (Thamnophis sirtalis parietalis) as mesocercariae of Alaria marcianae, Alaria mustelae, and Alaria sp. as well as metacercariae of Diplostomidae sp. of unknown generic affiliation. We assessed infection prevalence, absolute and relative intensity, and associated pathological changes in these snakes. Infection prevalence was 100 % for both male and female snakes. Infection intensity ranged from 11 to more than 2000 mesocercariae per snake tail but did not differ between the sexes. Gross pathological changes included tail swelling while histopathological changes included mild inflammation and the presence of mucus-filled pseudocysts surrounding mesocercariae, as well as the compression and degeneration of muscle fibers. Our results indicate that mesocercariae can lead to extensive muscle damage and loss in both sexes which likely increases the fragility of the tail making it more prone to breakage. As tail loss in garter snakes can affect both survival and reproduction, infection by Alaria mesocercariae clearly has serious fitness implications for these snakes.
Miyamoto, Kazuko; Kirinoki, Masashi; Matsuda, Hajime; Hayashi, Naoko; Chigusa, Yuichi; Sinuon, Muth; Chuor, Char Meng; Kitikoon, Viroj
Opisthorchiasis is endemic in Thailand and Lao People's Democratic Republic and constitutes a major public health problem throughout the Mekong Basin. Although Cambodia is located in the Mekong Basin, the status of O. viverrini infection in that country was not previously clarified. This research was conducted to document the extent and distribution of O. viverrini infection in Cambodia. Surveillance was conducted in 55 villages in five Cambodian provinces. Research tools included stool examination using the Kato-Katz thick-smear technique, identification of intermediate hosts, and interviews covering factors related to O. viverrini infection. Some larvae and egg-positive stool samples were examined using PCR to detect O. viverrini DNA. A total of 16,082 stool samples from the 55 villages were examined, of which 1232 were egg positive. In 15 villages with egg-positive rates of greater than 10%, eggs were found in 998 of 3585 stool samples, for an egg-positive rate of 27.8%. PCR analysis showed that 30 of 33 samples were positive for O. viverrini DNA from five villages in Kampong Cham and Kampong Thom provinces. The first intermediate host Bithynia siamensis siamensis was identified in the target areas of Takaev, Kandal, and Kampong Cham provinces. Cercariae were identified morphologically as O. viverrini and some were confirmed using PCR. Metacercariae of O. viverrini were identified by morphologic observations, animal experiments, or PCR in six species of fish in the target areas. Four Cambodian provinces were identified as endemic areas of O. viverrini infection. Careful planning is necessary for effective field surveys, because complex environmental factors might be involved in the distribution of O. viverrini infection-endemic areas in Cambodia. Many problems remain to be resolved regarding the status of O. viverrini infection in Cambodia, and a nationwide baseline survey is necessary. Copyright © 2013 Elsevier B.V. All rights reserved.
Relação entre fator de condição relativo (Kn e abundância de ectoparasitos de brânquias, em duas espécies de ciclídeos da bacia do rio Paraná, Brasil - DOI: 10.4025/actascibiolsci.v30i2.3625 Relation between the relative condition factor (Kn and the abundance of gill ectoparasites in two species of cichlids from the Paraná river basin, Brazil - DOI: 10.4025/actascibiolsci.v30i3.3625
Gilberto Cezar Pavanelli
Full Text Available Foram analisados 33 espécimes de Satanoperca pappaterra, capturados entre março de 2004 e junho de 2005, na planície de inundação do alto rio Paraná e 33 espécimes de Crenicichla niederleinii, capturados entre novembro de 2005 e novembro de 2006, no reservatório de Itaipu. Ambos estavam parasitados por monogenéticos, pertencentes ao gênero Sciadicleithrum e por metacercárias de Ascocotyle sp. O fator de condição relativo (Kn não diferiu significativamente entre indivíduos parasitados e não-parasitados das duas espécies de hospedeiros. Apenas Sciadicleithrum sp.1 parasito de S. pappaterra apresentou correlação positiva e significativa entre o fator de condição relativo (Kn e a abundância de parasitismo. Por outro lado, Ascocotyle sp. de ambos os hospedeiros e Sciadicleithrum sp.2 parasito de C. niederleinii não apresentaram correlações entre o Kn e a abundância de parasitismo.The study analyzed 33 specimens of Satanoperca pappaterra, captured from the upper Paraná river floodplain between March 2004 and June of 2005, and 33 specimens of Crenicichla niederleinii captured from the Itaipu Reservoir between November 2005 and November 2006. Both were parasitized by monogeneans pertaining to the genus Sciadicleithrum and by metacercariae of Ascocotyle sp. The relative condition factor (Kn did not differ significantly between parasitized and unparasitized individuals from the two host species. Only Sciadicleithrum sp.1, parasite of S. pappaterra, presented positive and significant correlation between the relative condition factor (Kn and the abundance of parasitism. On the other hand, Ascocotyle sp. in both hosts and Sciadicleithrum sp.2 parasite of C. niederleinii did not present correlations between the Kn and the abundance of parasitism.
Relação entre fator de condição relativo (Kn e abundância de ectoparasitos de brânquias, em duas espécies de ciclídeos da bacia do rio Paraná, Brasil = Relation between the relative condition factor (Kn and the abundance of gill ectoparasites in two species of cichlids from the Paraná river basin, Brazil
Fábio Hideki Yamada
Full Text Available Foram analisados 33 espécimes de Satanoperca pappaterra, capturados entre março de 2004 e junho de 2005, na planície de inundação do alto rio Paraná e 33 espécimes de Crenicichla niederleinii, capturados entre novembro de 2005 e novembro de 2006, no reservatório de Itaipu. Ambos estavam parasitados por monogenéticos, pertencentes ao gênero Sciadicleithrum e por metacercárias de Ascocotyle sp. O fator de condição relativo (Kn não diferiu significativamente entre indivíduos parasitados e não-parasitados das duas espécies de hospedeiros. Apenas Sciadicleithrum sp.1 parasito de S. pappaterra apresentou correlação positiva e significativa entre o fator de condição relativo (Kn e a abundância de parasitismo. Por outro lado, Ascocotyle sp. de ambos os hospedeiros e Sciadicleithrum sp.2parasito de C. niederleinii não apresentaram correlações entre o Kn e a abundância de parasitismo.The study analyzed 33 specimens of Satanoperca pappaterra, captured from the Upper Paraná River floodplain between March 2004 and June of 2005, and 33 specimens of Crenicichla niederleinii captured from the Itaipu Reservoir between November 2005 and November 2006. Both were parasitized by monogeneans pertaining to the genus Sciadicleithrum and by metacercariae of Ascocotyle sp. The relative condition factor (Kn did not differ significantly between parasitized and unparasitized individuals from the two host species. Only Sciadicleithrum sp.1, parasite of S. pappaterra, presented positive and significant correlation between the relative condition factor (Kn and the abundance of parasitism. On the other hand, Ascocotyle sp. in both hosts and Sciadicleithrum sp.2 parasite of C. niederleiniidid not present correlations between the Kn and the abundance of parasitism.
Sripa, Jittiyawadee; Laha, Thewarach; Sripa, Banchob
In the present study, the cDNA encoding FABP (Ov-FABP) was isolated from the adult stage of Opisthorchis viverrini and characterized. The Ov-FABP protein sequence (107 amino acids) was predicted to have a molecular mass of 12.26kDa and an isoelectric point (PI) of 6.82. This sequence had a high identity and similarity to Cs-FABP of the related opisthorchid Clonorchis sinensis. Multiple sequence alignment with FABPs from other parasitic flatworms and mammals showed a number of conserved amino acids including Phe 34 , Gly 37 , Glu 38 , Glu 39 ,Val 50 , Iso 62 , Gly 81 , Ile 84 , Ser 87 and Arg 101 . In addition, the structure of Ov-FABP was predicted to have eleven β-sheets and one α-helix based on the known structures for FABPs from human (hL-FABP), rat and a schistosome. Phylogenetic analysis of amino acid sequence data revealed a close relationship of Ov-FABP with Cs-FABP and hL-FABP. Reverse transcription-PCR revealed that Ov-FABP was transcribed in the egg, metacercaria, juvenile and adult stages. The soluble form of recombinant Ov-FABP (rOv-FABP) was shown to specifically bind fatty acids, including oleic acid, palmitic acid and linoleic acid, as shown for other animals. Anti-serum against rOv-FABP (produced in mice) located the protein to parenchyma, egg, sucker musculature, testes and tegument of adult O. viverrini. Taken together, the findings suggest key functional roles for Ov-FABP in development, reproduction and/or host-parasite interactions. Copyright © 2016 The Authors. Published by Elsevier B.V. All rights reserved.
Mahmoud, Mahmoud A; Abdelsalam, Mohamed; Mahdy, Olfat A; El Miniawy, Hala M F; Ahmed, Zakia A M; Osman, Ahmed H; Mohamed, Hussein M H; Khattab, A M; Zaki Ewiss, M A
This paper is a part of a multi-disciplinary research "Application of Decentralized On-Site Water Treatment System in Egypt for Use in Agriculture and Producing Safe Fish and Animal Proteins". The project aimed to investigate the environmental impact of implementing sewage water before and after treatment using the effluent of the on-site decentralized Japanese' Johkasou system, in agriculture and producing fish protein. The aim is to establish such system in Egypt to strengthen the sanitary conditions of water resources. In the present study, the impact of the sewage pollution in some fish farms at El-Fayyum, Port Said and El-Dakahlia governorates in Egypt was carried out. Water and fish (Oreochromis niloticus and Mugil cephalus) samples were collected from private fish farms of such localities. Bacteriological and chemical examination of water samples revealed the existence of coliforms and many other bacterial species of significant human health hazards. The chemical parameters of water showed a marked deviation from normal levels while examination of fish flesh specimens indicated contamination with Streptococcus Sp., Staphylococcus Sp., and Salmonella in all examined localities. Other bacterial isolates of human health importance (Morganella morganii, Pseudomonas cepacia and Enterococcos durans) were identified. The parasitological examination revealed the presence of encysted metacercariae (EMC); Diplostomatidae, Prohemistomatidae and Heterphyidae. Moreover, two protozoan parasites (Mxyoboulus tilapiae and Ichthyophthirius multifilis) were also recorded. The histopathological examination revealed mild tissue reaction in case of bacterial infection and severe pathological lesions in different organs in case of EMC infection. Lamellar hyperplasia and mononuclear cell infiltration in branchial tissue was common findings. In skeletal muscles, atrophy of muscle fibres, myolysis and myophagia were detected. Copyright © 2016 Elsevier Ltd. All rights reserved.
Rehman, Abdur; Ullah, Rizwan; Jaiswal, Neeshma; Khan, M A Hannan; Rehman, Lubna; Beg, Mirza Ahmar; Malhotra, Sandeep K; Abidi, S M A
The helminth parasites possess great capabilities to adapt themselves within their hosts and also develop strategies to render the commonly used anthelmintics ineffective leading to the development of resistance against these drugs. Besides using anthelmintics the natural products have also been tested for their anti-parasitic effects. Therapeutic efficacy of honey bee venom (HBV) has been tested in various ailments including some protozoal infections but very little is known about its anthelmintic properties. To investigate the anthelmintic effect of HBV the excysted progenetic metacercariae of Clinostomum complanatum, a heamophagic, digenetic trematode with zoonotic potential, infecting a wide variety of hosts, were obtained from Trichogaster fasciatus, a forage fish, which serves as the intermediate host. The metacercarial worms were in vitro incubated in RPMI-1640 medium containing HBV along with the controls which were devoid of HBV for the analysis of worm motility, enzyme activity, polypeptide profile and surface topographical changes. The motility of the worms was significantly reduced in a time dependent manner with an increase in the concentration of HBV. Following incubation of worms the release of cysteine proteases was inhibited in the presence of HBV as revealed by gelatine substrate gel zymography. As well as the polypeptide profile was also significantly influenced, particularly intensity/expression of M r 19.4 kDa, 24 kDa and 34 kDa was significantly reduced upon HBV treatment. The HBV treatment also inhibited antioxidant enzyme, superoxide dismutase (SOD) and Glutathione-S-transferase (GST) significantly (p < 0.05) in the worms. The scanning electron microscopy of the HBV treated worms revealed tegumental disruptions and erosion of papillae as well as spines showing vacuolation in the tegument. The HBV treated worms also showed a marked decline in the transformation rate when introduced into an experimental host which further reflect the
Donald Arguedas Cortés
Full Text Available Centrocestus formosanus es un parásito trematodo zoonótico originario de Asia asociado con muertes de peces principalmente de cultivo. 907 moluscos provenientes de estanques sembrados con tilapias, seleccionados uno por provincia fueron identificados al nivel taxonómico especifico. Se identificaron cuatro gastrópodos y un bivalvo: M. tuberculata, M. turricula, P. flagellata, H. cubensis y A. luteola. Se reporta, por primera vez, la presencia de dos especies de moluscos en Costa Rica. Se identificaron siete morfotipos de cercarias parasitando las cinco especies de moluscos encontradas. En la segunda exposición experimental se demostró que el morfotipo parapleurolofocercus encontrado en M. tuberculata concuerda con el hallazgo de C. formosanus en alevines de tilapia, después del examen clínico, anatomopatológico y parasitológico realizado a los alevines expuestos. Las metacercarias fueron extraídas del quiste utilizando microagujas y micropinzas lavadas en solución salina fisiológica (0.65%, fijadas en formol caliente al 4% y después esquematizadas con una cámara clara adaptada a un microscopio fotónico, estimándose una abundancia e intensidad media de 1018-1027 digeneos por branquia en cada pez parasitado, determinándose así el hospedador intermediario primario y secundario del parásito. En el presente trabajo se reporta por primera vez Centrocestus formosanus en Costa Rica.
Escamilla, A; Pérez-Caballero, R; Zafra, R; Bautista, M J; Pacheco, I L; Ruiz, M T; Martínez-Cruz, M S; Martínez-Moreno, A; Molina-Hernández, V; Pérez, J
Several immunomodulatory properties have been described in Fasciola hepatica infections. Apoptosis has been shown to be an effective mechanism to avoid the immune response in helminth infections. The aim of the present work was to study apoptosis in peritoneal leucocytes of sheep experimentally infected with F. hepatica during the early stages of infection. Five groups (n=5) of sheep were used. Groups 2-5 were orally infected with 200 metacercariae (mc) and sacrificed at 1, 3, 9 and 18days post-infection (dpi), respectively. Group 1 was used as the uninfected control (UC). Apoptosis was detected using three different methods 1) immunocytochemistry (ICC) with a polyclonal antibody anti-active caspase-3; 2) an annexin V flow cytometry assay using the Annexin V-FITC/propidium iodide (PI); and 3) transmission electron microscopy (TEM). The differential leucocyte count revealed that the majority of peritoneal granulocytes were eosinophils, which increased significantly at 9 and 18 dpi with respect to the uninfected controls. The ICC study revealed that the percentage of caspase-3 + apoptotic peritoneal leucocytes increased significantly from 3 dpi onwards with respect to the uninfected controls. The flow cytometry annexin V assay detected a very significant (P<0.001) increase of apoptotic peritoneal macrophages, lymphocytes and granulocytes, which remained higher than in the UC until 18 dpi. Transmission electron microscopy studies also confirmed the presence of apoptosis in peritoneal eosinophils at 18 dpi. This is the first report of apoptosis induced by F. hepatica in the peritoneal leucocytes of sheep in vivo. The results of this work suggest the importance of apoptosis induction for the survival of the juvenile parasites in the peritoneal migratory stages of infection. Copyright © 2017 Elsevier B.V. All rights reserved.
Full Text Available Fasciolosis is considered the most widespread trematode disease affecting grazing animals around the world; it is currently recognised by the World Health Organisation as an emergent human pathogen. Triclabendazole is still the most effective drug against this disease; however, resistant strains have appeared and developing an effective vaccine against this disease has increasingly become a priority. Several bioinformatics tools were here used for predicting B- and T-cell epitopes according to the available data for Fasciola hepatica protein amino acid sequences. BALB/c mice were immunised with the synthetic peptides by using the ADAD vaccination system and several immune response parameters were measured (antibody titres, cytokine levels, T-cell populations to evaluate their ability to elicit an immune response. Based on the immunogenicity results so obtained, seven peptides were selected to assess their protection-inducing ability against experimental infection with F. hepatica metacercariae. Twenty-four B- or T-epitope-containing peptides were predicted and chemically synthesised. Immunisation of mice with peptides so-called B1, B2, B5, B6, T14, T15 and T16 induced high levels of total IgG, IgG1 and IgG2a (p<0.05 and a mixed Th1/Th2/Th17/Treg immune response, according to IFN-γ, IL-4, IL-17 and IL-10 levels, accompanied by increased CD62L+ T-cell populations. A high level of protection was obtained in mice vaccinated with peptides B2, B5, B6 and T15 formulated in the ADAD vaccination system with the AA0029 immunomodulator. The bioinformatics approach used in the present study led to the identification of seven peptides as vaccine candidates against the infection caused by Fasciola hepatica (a liver-fluke trematode. However, vaccine efficacy must be evaluated in other host species, including those having veterinary importance.
Maggioli, Gabriela; Bottini, Gualberto; Basika, Tatiana; Alonzo, Pablo; Salinas, Gustavo; Carmona, Carlos
The liver fluke Fasciola hepatica remains an important agent of food-borne trematode disease producing great economic losses due to its negative effect on productivity of livestock grazing in temperate areas. The prevailing control strategy based on anthelmintic drugs is unsustainable due to widespread resistance hence vaccination appears as an attractive option to pursue. In this study we evaluate the effect of vaccination in calves with a functional recombinant thioredoxin glutathione reductase (rFhTGR) from liver fluke, a critical antioxidant enzyme at the crossroads of the thioredoxin and glutathione metabolism in flatworms. The recombinant enzyme produced in Escherichia coli was tested in two vaccination experiments; in the first trial rFhTGR was administered in combination with Freund́s Incomplete Adjuvant (FIA) in a three-inoculation scheme on weeks 0, 4 and 8; in the second trial rFhTGR was given mixed with Adyuvac 50 or Alum as adjuvants on weeks 0 and 4. In both cases calves were challenged with metacercariae (400 in the first and 500 in the second trial) 2 weeks after the last inoculation. Our results demonstrate that two or three doses of the vaccine induced a non-significant reduction in worm counts of 8.2% (FIA), 3.8% (Adyuvac 50) and 23.0% (Alum) compared to adjuvant controls indicating that rFhTGR failed to induce a protective immunity in challenged calves. All vaccine formulations induced a mixed IgG1/IgG2 response but no booster was observed after challenge. No correlations between antibody titres and worm burdens were found. Copyright © 2016 Elsevier B.V. All rights reserved.
Cwiklinski, Krystyna; Allen, Katherine; LaCourse, James; Williams, Diana J; Paterson, Steve; Hodgkinson, Jane E
The liver fluke, Fasciola hepatica is an economically important pathogen of sheep and cattle and has been described by the WHO as a re-emerging zoonosis. Control is heavily reliant on the use of drugs, particularly triclabendazole and as a result resistance has now emerged. The population structure of F. hepatica is not well known, yet it can impact on host-parasite interactions and parasite control with drugs, particularly regarding the spread of triclabendazole resistance. We have identified 2448 potential microsatellites from 83 Mb of F. hepatica genome sequence using msatfinder. Thirty-five loci were developed and optimised for microsatellite PCR, resulting in a panel of 15 polymorphic loci, with a range of three to 15 alleles. This panel was validated on genomic DNA from 46 adult F. hepatica; 38 liver flukes sourced from a Northwest abattoir, UK and 8 liver flukes from an established isolate (Shrewsbury; Ridgeway Research). Evidence for null alleles was found at four loci (Fh_1, Fh_8, Fh_13 and Fh_14), which showed markedly higher levels of homozygosity than the remaining 11 loci. Of the 38 liver flukes isolated from cattle livers (n=10) at the abattoir, 37 genotypes were identified. Using a multiplex approach all 15 loci could be amplified from several life cycle stages that typically yield low amounts of DNA, including metacercariae, the infective life cycle stage present on pasture, highlighting the utility of this multiplex microsatellite panel. This study reports the largest panel of microsatellite markers available to date for population studies of F. hepatica and the first multiplex panel of microsatellite markers that can be used for several life cycle stages. Copyright © 2015 The Authors. Published by Elsevier B.V. All rights reserved.
Rivera, Francheska; Espino, Ana M
Fasciola hepatica saposin-like protein-2 (FhSAP2) is a protein differentially expressed in various developmental stages of F. hepatica. Recombinant FhSAP2 has demonstrated the induction of partial protection in mice and rabbits when it is administered subcutaneously (SC) in Freund's adjuvant. Because FhSAP2 is overexpressed in bacteria in the form of inclusion bodies (IBs), we isolated IBs expressing FhSAP2 and tested their immunogenicity when administered SC in mice emulsified in two different adjuvants: QS-21 and Montanide TM ISA720. Animals received three injections containing 20 μg of protein two weeks apart and 4 weeks after the third injection, mice were infected with 10 F. hepatica metacercariae by oral route. The percentages of protection induced by FhSAP2-IBs were estimated to be between 60.0 and 62.5% when compared with adjuvant-vaccinated, infected controls. By determining the levels of IgG1 and IgG2a antibodies and IL-4 and IFNγ cytokines in the serum of experimental animals, it was found that both Th1 and Th2 immune responses were significantly increased in the FhSAP2-IBs vaccinated groups compared with the adjuvant-vaccinated, infected control groups. The adjuvant-vaccinated groups had significantly lower IgG1 to IgG2a ratios and lower IL-4 to IFNγ ratios than the FhSAP2-IBs vaccinated animals, which is indicative of higher levels of Th2 immune responses. Irrespective to the adjuvant used, animals vaccinated with FhSAP2-IBs exhibited significantly higher survival percentage and less liver damage than the adjuvant-control groups. This study suggests that FhSAP2 has potential as vaccine against F. hepatica and that the protection elicited by this molecule could be linked to a mechanism driven by the CD4-Th1 cells. Published by Elsevier Inc.
Gerardo Pérez-Ponce de León
Full Text Available A new species of Theletrum is described from the intestine of two palenose morays, Echidna nocturna, collected in Cuajiniquil, Guanacaste Province, Costa Rica. The new species differs from the type species, T. fustiforme Linton, 1910 by having a subspherical pars prostatica, a subspherical seminal vesicle extending anteriorly to the anterior border of the acetabulum, by the presence of a poorly developed hermaphroditic sac, and by having a larger body size. We also report eight additional species of digeneans parasitizing marine fishes in several localities along the Atlantic and Pacific coast of Costa Rica: Bianium simonei, Didymozoinae (metacercariae, Ectenurus virgulus, Hypocreadium myohelicatum, Lecithochirium microstomum, Pseudolecithaster sp., Stephanostomum casum, and Tergestia laticollis. In addition, we present an updated list of helminth parasites of marine fish from Costa Rica and discuss the importance of including parasites as an integral part of biodiversity inventories.Se describe una especie nueva de digéneo del género Theletrum, parásito del intestino de una "morena pecosa", Echidna nocturna, recolectada en Playa Cuajiniquil, Provincia de Guanacaste, Costa Rica. La nueva especie es diferente de T. fustiforme Linton, 1910, la especie tipo, por presentar una pars prostatica subesférica, una vesícula seminal subesférica que se extiende hasta el borde anterior del acetábulo, por la presencia de un saco hermafrodita y por una mayor longitud del cuerpo. También se registran ocho especies adicionales de digéneos parásitos de peces marinos en varias localidades de las costas del Atlántico y del Pacífico de Costa Rica. Por último, presentamos una lista actualizada de los helmintos parásitos de peces marinos y dulceacuícolas de Costa Rica y discutimos la importancia de incluir a los parásitos como una parte integral de las investigaciones sobre biodiversidad.
Rojas-Caraballo, Jose; López-Abán, Julio; Pérez del Villar, Luis; Vizcaíno, Carolina; Vicente, Belén; Fernández-Soto, Pedro; del Olmo, Esther; Patarroyo, Manuel Alfonso; Muro, Antonio
Fasciolosis is considered the most widespread trematode disease affecting grazing animals around the world; it is currently recognised by the World Health Organisation as an emergent human pathogen. Triclabendazole is still the most effective drug against this disease; however, resistant strains have appeared and developing an effective vaccine against this disease has increasingly become a priority. Several bioinformatics tools were here used for predicting B- and T-cell epitopes according to the available data for Fasciola hepatica protein amino acid sequences. BALB/c mice were immunised with the synthetic peptides by using the ADAD vaccination system and several immune response parameters were measured (antibody titres, cytokine levels, T-cell populations) to evaluate their ability to elicit an immune response. Based on the immunogenicity results so obtained, seven peptides were selected to assess their protection-inducing ability against experimental infection with F. hepatica metacercariae. Twenty-four B- or T-epitope-containing peptides were predicted and chemically synthesised. Immunisation of mice with peptides so-called B1, B2, B5, B6, T14, T15 and T16 induced high levels of total IgG, IgG1 and IgG2a (ppeptides B2, B5, B6 and T15 formulated in the ADAD vaccination system with the AA0029 immunomodulator. The bioinformatics approach used in the present study led to the identification of seven peptides as vaccine candidates against the infection caused by Fasciola hepatica (a liver-fluke trematode). However, vaccine efficacy must be evaluated in other host species, including those having veterinary importance. PMID:25122166
Rojas-Caraballo, Jose; López-Abán, Julio; Pérez del Villar, Luis; Vizcaíno, Carolina; Vicente, Belén; Fernández-Soto, Pedro; del Olmo, Esther; Patarroyo, Manuel Alfonso; Muro, Antonio
Fasciolosis is considered the most widespread trematode disease affecting grazing animals around the world; it is currently recognised by the World Health Organisation as an emergent human pathogen. Triclabendazole is still the most effective drug against this disease; however, resistant strains have appeared and developing an effective vaccine against this disease has increasingly become a priority. Several bioinformatics tools were here used for predicting B- and T-cell epitopes according to the available data for Fasciola hepatica protein amino acid sequences. BALB/c mice were immunised with the synthetic peptides by using the ADAD vaccination system and several immune response parameters were measured (antibody titres, cytokine levels, T-cell populations) to evaluate their ability to elicit an immune response. Based on the immunogenicity results so obtained, seven peptides were selected to assess their protection-inducing ability against experimental infection with F. hepatica metacercariae. Twenty-four B- or T-epitope-containing peptides were predicted and chemically synthesised. Immunisation of mice with peptides so-called B1, B2, B5, B6, T14, T15 and T16 induced high levels of total IgG, IgG1 and IgG2a (ppeptides B2, B5, B6 and T15 formulated in the ADAD vaccination system with the AA0029 immunomodulator. The bioinformatics approach used in the present study led to the identification of seven peptides as vaccine candidates against the infection caused by Fasciola hepatica (a liver-fluke trematode). However, vaccine efficacy must be evaluated in other host species, including those having veterinary importance.
El-Ahwany, Eman; Rabia, Ibrahim; Nagy, Faten; Zoheiry, Mona; Diab, Tarek; Zada, Suher
Fascioliasis is one of the public health problems in the world. Cysteine proteinases (CP) released by Fasciola gigantica play a key role in parasite feeding, migration through host tissues, and in immune evasion. There has been some evidence from several parasite systems that proteinases might have potential as protective antigens against parasitic infections. Cysteine proteinases were purified and tested in vaccine trials of sheep infected with the liver fluke. Multiple doses (2 mg of CP in Freund's adjuvant followed by 3 booster doses 1 mg each at 4 week intervals) were injected intramuscularly into sheep 1 week prior to infect orally with 300 F. gigantica metacercariae. All the sheep were humanely slaughtered 12 weeks after the first immunization. Changes in the worm burden, ova count, and humoral and cellular responses were evaluated. Significant reduction was observed in the worm burden (56.9%), bile egg count (70.7%), and fecel egg count (75.2%). Immunization with CP was also found to be associated with increases of total IgG, IgG(1), and IgG(2) (P<0.05). Data showed that the serum cytokine levels of pro-inflammatory cytokines, IL-12, IFN-γ, and TNF-α, revealed significant decreases (P<0.05). However, the anti-inflammatory cytokine levels, IL-10, TGF-β, and IL-6, showed significant increases (P<0.05). In conclusion, it has been found that CP released by F. gigantica are highly important candidates for a vaccine antigen because of their role in the fluke biology and host-parasite relationships.
Huang, Yanwei; Zheng, Youwei; Li, Yuzhe; Yang, Mei; Li, Ting; Zeng, Suxiang; Yu, Xinbing; Huang, Huaiqiu; Hu, Xuchu
Clonorchiasis, caused by Clonorchis sinensis infection, is a zoonotic parasitic disease of hepatobiliary system in which the proteins released by adult are major pathogenetic factors. In this study, we first characterized a putative sphingomyelin phosphodiesterase (CsSMPase) A-like secretory protein, which was highly expressed in the adult worm. The full-length gene was cloned. The putative protein is of relatively low homology comparing with SMPase from other species, and of rich T cell and B cell epitopes, suggesting that it is an antigen of strong antigenicity. The complete coding sequence of the gene was expressed in the Escherichia coli. The recombinant CsSMPase (rCsSMPase) can be recognized by C. sinensis-infected serum, and the protein immunoserum can recognize a specific band in excretory/secretory products (ESPs) of C. sinensis adult by western blotting. Immunolocalization revealed that CsSMPase was not only localized on tegument, ventral sucker of metacercaria, and the intestine of adult but also on the nearby epithelium of bile duct of the infected Sprague-Dawley rats, implying that CsSMPase was mainly secreted and excreted through adult intestine and directly interacted with bile duct epithelium. Although immunized rats evoked high level antibody response, the antigen level was low in clonorchiasis patients. And the sensitivity and specificity of rCsSMPase were 50.0 % (12/24) and 88.4 % (61/69), in sera IgG-ELISA, respectively. It is likely due to the fact that CsSMPase binding to the plasma membrane of biliary epithelium decreases the antigen immune stimulation.
Full Text Available BACKGROUND: Adult Clonorchis sinensis lives in the bile duct and causes endemic clonorchiasis in East Asian countries. Phosphagen kinases (PK constitute a highly conserved family of enzymes, which play a role in ATP buffering in cells, and are potential targets for chemotherapeutic agents, since variants of PK are found only in invertebrate animals, including helminthic parasites. This work is conducted to characterize a PK from C. sinensis and to address further investigation for future drug development. METHODOLOGY/PRINCIPAL FINDINGS: [corrected] A cDNA clone encoding a putative polypeptide of 717 amino acids was retrieved from a C. sinensis transcriptome. This polypeptide was homologous to taurocyamine kinase (TK of the invertebrate animals and consisted of two contiguous domains. C. sinensis TK (CsTK gene was reported and found consist of 13 exons intercalated with 12 introns. This suggested an evolutionary pathway originating from an arginine kinase gene group, and distinguished annelid TK from the general CK phylogenetic group. CsTK was found not to have a homologous counterpart in sequences analysis of its mammalian hosts from public databases. Individual domains of CsTK, as well as the whole two-domain enzyme, showed enzymatic activity and specificity toward taurocyamine substrate. Of the CsTK residues, R58, I60 and Y84 of domain 1, and H60, I63 and Y87 of domain 2 were found to participate in binding taurocyamine. CsTK expression was distributed in locomotive and reproductive organs of adult C. sinensis. Developmentally, CsTK was stably expressed in both the adult and metacercariae stages. Recombinant CsTK protein was found to have low sensitivity and specificity toward C. sinensis and platyhelminth-infected human sera on ELISA. CONCLUSION: CsTK is a promising anti-C. sinensis drug target since the enzyme is found only in the C. sinensis and has a substrate specificity for taurocyamine, which is different from its mammalian counterpart
Uhrig, Emily J.; Spagnoli, Sean T.; Tkach, Vasyl V.; Kent, Michael L.; Mason, Robert T.
Trematodes of the genus Alaria develop into an arrested stage, known as mesocercariae, within their amphibian second intermediate host. The mesocercariae are frequently transmitted to a non-obligate paratenic host before reaching a definitive host where further development and reproduction can occur. Snakes are common paratenic hosts for Alaria spp. with the mesocercariae often aggregating in the host’s tail. In the current study, we used morphological examination and molecular analyses based on partial sequences of nuclear large ribosomal subunit gene and mitochondrial cytochrome C oxidase subunit 1 gene to identify larvae in the tails of red-sided garter snakes (Thamnophis sirtalis parietalis) as mesocercariae of Alaria marcianae, Alaria mustelae, and Alaria sp. as well as metacercariae of Diplostomidae sp. of unknown generic affiliation. We assessed infection prevalence, absolute and relative intensity, and associated pathological changes in these snakes. Infection prevalence was 100% for both male and female snakes. Infection intensity ranged from 11 to more than 2,000 mesocercariae per snake tail, but did not differ between the sexes. Gross pathological changes included tail swelling while histopathological changes included mild inflammation and the presence of mucus-filled pseudocysts surrounding mesocercariae, as well as the compression and degeneration of muscle fibers. Our results indicate that mesocercariae can lead to extensive muscle damage and loss in both sexes which likely increases the fragility of the tail making it more prone to breakage. As tail loss in garter snakes can affect both survival and reproduction, infection by Alaria mesocercariae clearly has serious fitness implications for these snakes. PMID:26337267
Stunkard, H.W.; Uzmann, J.R.
The genus Proctoeces was erected by Odhner ( 191 1) to contain Distonium maculatuni Looss, 1901, from Labrus merula and Crenilabrus spp. at Triest. Odhner had found the parasite in Blennius ocellaris at Naples. One adult specimen from Chrysophrys bifasciata and two immature specimens from lulis lunaris taken in the Red Sea, were described as a new species, Proctoeces erythraeus. Dawes (1946) listed P. erythraeus as a synonym of P. maculatus (Looss) , but the species was recognized by Manter ( 1947) on the basis of six specimens he had collected from Calamus calamus and Calamus bajonado at the biological laboratory of the Carnegie Institution at Dry Tortugas, Florida. Several additional species have been de scribed. Fujita ( 1925) reported a metacercaria from the Japanese oyster, Ostrea gigas, as a new species, Proctoeces ostreae. The paper was translated by R. Ph. Dollfus who noted (p. 57) ,“Il est à souhaiter que des recherches chez les poissons mangers de Lamellibranches, sur les côtes de la préfecture d'Hiroshima, permettent de découvrir des exemplaires complètement adultes de Proctoeces ostreae Fuj., chez lesquels l'extension des vitellogènes et les dimensions des oeufs puissent être observées avec précision; il sera alors possible de savoir définitivement si P. ostreae Fuj. doit ou non tomber en synonymie avec P. maculatus (Looss)." Yamaguti (1934) described P. maculatus from Sparus aries, Sparws macrocephalus, Pagrosomus auratus, and Epinephelus akaara in Japan. Several specimens from Pagrosomus auratus, which differed from P. maculatus in larger size, larger eggs, and trilobed ovary, he described as a new species, Proctoeces major. Yamaguti ( 1938) reported P. nzaculatus from Sensicossyphus reticulatus and described a larva from the liver of the pelecypod mollusk, Brachidontes senhausi, as an unidentified member of the genus Proctoeces. Manter ( 1940) described Proctoeces tnagnorus from a single specimen found in the intestine of Caulolatilus
Rojas-Caraballo, Jose; López-Abán, Julio; Moreno-Pérez, Darwin Andrés; Vicente, Belén; Fernández-Soto, Pedro; Del Olmo, Esther; Patarroyo, Manuel Alfonso; Muro, Antonio
Fasciolosis remains a significant food-borne trematode disease causing high morbidity around the world and affecting grazing animals and humans. A deeper understanding concerning the molecular mechanisms by which Fasciola hepatica infection occurs, as well as the molecular basis involved in acquiring protection is extremely important when designing and selecting new vaccine candidates. The present study provides a first report of microarray-based technology for describing changes in the splenic gene expression profile for mice immunised with a highly effective, protection-inducing, multi-epitope, subunit-based, chemically-synthesised vaccine candidate against F. hepatica. The mice were immunised with synthetic peptides containing B- and T-cell epitopes, which are derived from F. hepatica cathepsin B and amoebapore proteins, as novel vaccine candidates against F. hepatica formulated in an adjuvant adaptation vaccination system; they were experimentally challenged with F. hepatica metacercariae. Spleen RNA from mice immunised with the highest protection-inducing synthetic peptides was isolated, amplified and labelled using Affymetrix standardised protocols. Data was then background corrected, normalised and the expression signal was calculated. The Ingenuity Pathway Analysis tool was then used for analysing differentially expressed gene identifiers for annotating bio-functions and constructing and visualising molecular interaction networks. Mice immunised with a combination of three peptides containing T-cell epitopes induced high protection against experimental challenge according to survival rates and hepatic damage scores. It also induced differential expression of 820 genes, 168 genes being up-regulated and 652 genes being down-regulated, p value <0.05, fold change ranging from -2.944 to 7.632. A functional study of these genes revealed changes in the pathways related to nitric oxide and reactive oxygen species production, Interleukin-12 signalling and production
Full Text Available The tegument, representing the membrane-bound outer surface of platyhelminth parasites, plays an important role for the regulation of the host immune response and parasite survival. A comprehensive understanding of tegumental proteins can provide drug candidates for use against helminth-associated diseases, such as clonorchiasis caused by the liver fluke Clonorchis sinensis. However, little is known regarding the physicochemical properties of C. sinensis teguments. In this study, a novel 20.6-kDa tegumental protein of the C. sinensis adult worm (CsTegu20.6 was identified and characterized by molecular and in silico methods. The complete coding sequence of 525 bp was derived from cDNA clones and encodes a protein of 175 amino acids. Homology search using BLASTX showed CsTegu20.6 identity ranging from 29% to 39% with previously-known tegumental proteins in C. sinensis. Domain analysis indicated the presence of a calcium-binding EF-hand domain containing a basic helix-loop-helix structure and a dynein light chain domain exhibiting a ferredoxin fold. We used a modified method to obtain the accurate tertiary structure of the CsTegu20.6 protein because of the unavailability of appropriate templates. The CsTegu20.6 protein sequence was split into two domains based on the disordered region, and then, the structure of each domain was modeled using I-TASSER. A final full-length structure was obtained by combining two structures and refining the whole structure. A refined CsTegu20.6 structure was used to identify a potential CsTegu20.6 inhibitor based on protein structure-compound interaction analysis. The recombinant proteins were expressed in Escherichia coli and purified by nickel-nitrilotriacetic acid affinity chromatography. In C. sinensis, CsTegu20.6 mRNAs were abundant in adult and metacercariae, but not in the egg. Immunohistochemistry revealed that CsTegu20.6 localized to the surface of the tegument in the adult fluke. Collectively, our results
Kang, Jung-Mi; Lee, Jinyoung; Ju, Hye-Lim; Ju, Jung Won; Kim, Jong-Hyun; Pak, Jhang Ho; Kim, Tong-Soo; Hong, Yeonchul; Sohn, Woon-Mok; Na, Byoung-Kuk
Asparaginyl endopeptidases (AEP: EC 22.214.171.124) are a family of cysteine proteases classified into the MEROPS clan CD, family C13. In this study, we characterized the biochemical and antigenic properties of an AEP of Clonorchis sinensis (CsAEP). The recombinant CsAEP showed hydrolytic activity at pH values ranging from acidic to neutral with optimum activity at pH 6.0. While the recombinant CsAEP was stable at neutral pHs, it was unstable at acidic pHs and resulted in loss of enzymatic activity. The recombinant enzyme was effectively inhibited by iodoacetic acid and N-ethylmaleimide, but not by E-64. The partially purified native CsAEP showed biochemical properties similar to the recombinant enzyme. Native CsAEP is likely to be cleaved into an N-terminal mature enzyme and a C-terminal fragment via autocatalytic activation at acidic pHs. Polyclonal antibody raised against the recombinant CsAEP recognized three forms of CsAEP, proenzyme, the N-terminal mature enzyme and the C-terminal fragment, in the worm extract (WE) of C. sinensis. However, only the C-terminal fragment was mainly found in the excretory and secretory (ES) products of the parasite. Strong CsAEP activity was found in the WE, but only a trace level of CsAEP activity was detected in the ES products of the parasite. CsAEP was expressed in various developmental stages of C. sinensis, from metacercariae to adults, and was found to be localized in the intestine of the parasite as well as in intestinal contents. Sera from rats experimentally infected with C. sinensis reacted with CsAEP beginning 4 weeks after infection. These results suggest that CsAEP is a gut-associated enzyme synthesized in the intestine of C. sinensis and subsequently secreted into the intestinal lumen of the parasite. Copyright © 2015 Elsevier Inc. All rights reserved.
Kim, Yu-Jung; Yoo, Won Gi; Lee, Myoung-Ro; Kang, Jung-Mi; Na, Byoung-Kuk; Cho, Shin-Hyeong; Park, Mi-Yeoun; Ju, Jung-Won
The tegument, representing the membrane-bound outer surface of platyhelminth parasites, plays an important role for the regulation of the host immune response and parasite survival. A comprehensive understanding of tegumental proteins can provide drug candidates for use against helminth-associated diseases, such as clonorchiasis caused by the liver fluke Clonorchis sinensis . However, little is known regarding the physicochemical properties of C. sinensis teguments. In this study, a novel 20.6-kDa tegumental protein of the C. sinensis adult worm (CsTegu20.6) was identified and characterized by molecular and in silico methods. The complete coding sequence of 525 bp was derived from cDNA clones and encodes a protein of 175 amino acids. Homology search using BLASTX showed CsTegu20.6 identity ranging from 29% to 39% with previously-known tegumental proteins in C. sinensis . Domain analysis indicated the presence of a calcium-binding EF-hand domain containing a basic helix-loop-helix structure and a dynein light chain domain exhibiting a ferredoxin fold. We used a modified method to obtain the accurate tertiary structure of the CsTegu20.6 protein because of the unavailability of appropriate templates. The CsTegu20.6 protein sequence was split into two domains based on the disordered region, and then, the structure of each domain was modeled using I-TASSER. A final full-length structure was obtained by combining two structures and refining the whole structure. A refined CsTegu20.6 structure was used to identify a potential CsTegu20.6 inhibitor based on protein structure-compound interaction analysis. The recombinant proteins were expressed in Escherichia coli and purified by nickel-nitrilotriacetic acid affinity chromatography. In C. sinensis , CsTegu20.6 mRNAs were abundant in adult and metacercariae, but not in the egg. Immunohistochemistry revealed that CsTegu20.6 localized to the surface of the tegument in the adult fluke. Collectively, our results contribute to a
Ana M. Espino
Full Text Available En el presente estudio se describen cuáles son los antígenos de Fasciola hepatica que están presentes en las heces de pacientes con fascioliasis crónica y de ratas infectadas experimentalmente con metacercarias de F. hepatica. Mediante el empleo de la técnica de inmunoelectrotransferencia (Western blot con un suero hiperinmune obtenido frente a antígenos de excreción/secreción de adultos de F. hepatica, se pudo demostrar que en las heces de los pacientes se encuentran antígenos de posible interés para el diagnóstico con masas moleculares de 14, 19, 20, 23, 25, 32, 46, 51 y 62 kilodaltons (kDa. Además, se comprobó que algunos de estos péptidos (los de 14, 20, 23 y 51 kDa son reconocidos también por la mayoría de los sueros de pacientes crónicos. Después de purificar por cromatografía de afinidad los antígenos presentes en heces de ratas con 6 a 12 semanas de infección, empleando para ello el anticuerpo monoclonal ES78 acoplado a Sepharosa 4B-CNBr, se identificaron seis polipéptidos de 11, 14, 26, 32, 47 y 51 kDa; otros tres polipéptidos, de 17, 24 y 66 kDa, solo se pudieron identificar en heces de ratas con 10 a 12 semanas de infección. Nuestros resultados sugieren que estos polipéptidos pueden ser antígenos comunes a ambos estadios parasitarios y constituir importantes marcadores de la fascioliasis aguda y crónica, en particular los de 14, 24, 26 y 51 kDa, puesto que reaccionaron con el inmunosuero, los sueros humanos y el anticuerpo monoclonal ES78.
George, S D; Vanhoff, K; Baker, K; Lake, L; Rolfe, P F; Seewald, W; Emery, D L
At present diagnosis of true resistance and determination of drug efficacy in Fasciola hepatica infection rely solely on terminal experiments. The coproantigen ELISA (cELISA) has been reported previously as a sensitive and specific tool appropriate to detect treatment failure, and potentially drug resistance. Two studies were conducted to determine whether the cELISA was appropriate for on-farm efficacy and resistance testing in Australian Merino sheep. In Study 1 sheep were infected orally with 50 F. hepatica metacercariae on three occasions, twelve, six and two weeks prior to a single flukicide treatment with triclabendazole, closantel or albendazole. Sheep were sampled weekly for a further seven weeks prior to necropsy. Following effective treatment, no faecal antigen was detected from 1 week. When immature stages (≤6 weeks) survived treatment, coproantigen reappeared from 6 weeks post-treatment. Therefore, cELISA conducted 1-4 weeks after treatment will demonstrate obvious treatment failure against adult F. hepatica, but is not sufficiently sensitive to detect survival of immature fluke until these reach maturity. In study 2, fluke burdens of sheep necropsied 13 weeks post single infection were compared to fecal worm egg counts (FWEC) and cELISA at necropsy. Regression analysis demonstrated that cELISA correlated strongly with fluke burden, whilst FWEC correlated weakly with cELISA. The correlation between FWEC and fluke burden was also weak, although stronger than that of FWEC with cELISA. The cELISA is an appropriate tool for monitoring effectiveness of treatments against Fasciola hepatica if an adult infection is present, however when immature stages of the parasite are present it is not as reliable. Where immature parasites are present it is recommended that initial cELISA be followed with a secondary cELISA at least 6 weeks after treatment to ensure resistance to immature stages is detected. Further testing is justified for monitoring the effectiveness
Mawoad, M.A.F.; Amin, M.M.; Elnashar, D. E.
Fascioliasis is a zoonotic disease caused by the trematode Fasciola hepatica, a liver fluke. It affect numerous species mainly ruminants. Its usual hosts are sheep or cattle, in which it causes a disease of economic importance. The prevalence of human infection has been reported worldwide. The present study was designed to evaluate the comparative efficacy of different anthelmintic drugs against Fasciola hepatica infestation. Twenty parasite-free rabbits were divided into four groups, group I: non-infected non- treated rabbits, kept as negative control. Rabbits of group II, III and IV were intended for infection with 20 Fasciola hepatica metacercariae per animal. Group II received no treatment and kept as positive control. Animals of group III administered Triclabendazole (TCBZ) treatment orally in a single dose (0.45 mg/Kg b. wt.) 60 days after infection. Consistent with group III, animals of group IV administered a bendazole (ABZ) treatment orally in a single dose (0.52 mg/Kg b.wt.) The comparisons between the anthelmintic efficacy of TCBZ and ABZ were done throughout (1): parasitological evaluation of egg counts; (2): biochemical evaluation of serum AST and ALT; (3): histopathological examination of liver and (4): scanning electron microscope of host worms. Data represented showed that the egg count was highly significant reduced in rabbits treated with TCBZ as compared with those of positive control group. But, rabbits treated with ABZ revealed slightly reduction as compared with those of positive control group. Also, the results illustrated that infection of rabbits with Fasciola hepatica induced a highly significant increased in serum (AST and ALT) activities compared to control group. In response to treatment with TCBZ and ABZ, it was revealed a significant decrease in the enzyme activity of both serum AST and ALT levels compared to those of control negative group as well as control positive group. However, histopathological examination showed a significant
Ceballos, L; Canton, C; Cadenazzi, G; Larsen, K; Virkel, G; Moreno, L; Fairweather, I; Lanusse, C; Alvarez, L
Closantel (CLS) is highly effective against adult liver flukes after its oral or subcutaneous (sc) administration in ruminants. Trans-tegumental diffusion and oral ingestion are the two potential routes available for the entry of drugs into Fasciola hepatica. The work reported here contributes to improve the understanding of CLS pharmacology. The main goals of were: I) to determine the pattern of in vivo CLS accumulation into adult F. hepatica and relevant tissues in CLS-treated sheep; II) to investigate the influence of the physicochemical composition of the incubation medium on the CLS diffusion process into adult F. hepatica; III) to assess the ovicidal activity of CLS against F. hepatica eggs; and IV) to investigate the in vivo effect of CLS treatment on glutathione S-transferases activity in adult liver flukes exposed to CLS. Fourteen healthy sheep were each orally infected with 75 F. hepatica metacercariae. Sixteen (16) weeks after infection, animals were treated with CLS by oral (n = 6, 10 mg/kg) or sub-cutaneous (sc) (n = 6, 5 mg/kg) route. At 12, 24 and 36 h post-treatment, animals were sacrificed (n = 2) and samples of blood, bile and adult F. hepatica were collected. In addition, flukes recovered from non-treated sheep (n = 2) were ex vivo incubated (60 min) in the presence of CLS in either RPMI or bile as incubation medium. CLS concentration was measured by HPLC. The ovicidal activity of CLS was investigated using eggs obtained from the bile of untreated sheep. Finally, glutathione S-transferase activity in F. hepatica recovered from untreated and CLS-treated sheep was assessed. In the in vivo studies, the highest CLS concentrations were measured in plasma and adult liver flukes. A positive correlation was observed between CLS concentration in plasma and in F. hepatica. Results obtained in the current work indicate that the in vivo accumulation of CLS into adult liver flukes occurs mainly by the oral route. After ex
Sansri, Veerawat; Meemon, Krai; Changklungmoa, Narin; Kueakhai, Pornanan; Chantree, Pathanin; Chaichanasak, Pannigan; Lorsuwannarat, Natcha; Itagaki, Tadashi; Sobhon, Prasert
Fasciola gigantica cathepsin L1H (FgCatL1H) is one of the major cathepsin L released by juveniles of F. gigantica to aid in the invasion of host's tissues. Due to its high sequence similarity with other cathepsin L (CatL) isoforms of late stage F. gigantica, it was considered to be a good vaccine candidate that can block all CatL-mediated protease activities and affect juveniles as well as adult parasites. In this study, recombinant proFgCatL1H protein expressed in yeast, Pichia pastoris, system was mixed with Freund's adjuvants and used to subcutaneously immunize mice that were later challenged with metacercariae of F. gigantica. The percentage of worm protection in the rproFgCatL1H-vaccinated mice compared to the non-immunized and adjuvant control mice were approximately 62.7% and 66.1%, respectively. Anti-rproFgCatL1H antisera collected from vaccinated mice reacted specifically with rproFgCatL1H and other cathepsin L isoforms of F. gigantica, but the antibodies did not cross react with antigens from other trematode and nematode parasites, including Eurytrema pancreaticum, Opisthorchis viverrini, Fischoederius cobboldi, Cotylophoron cotylophorum, Gigantocotyle explanatum, Paramphistomum cervi, and Setaria labiato-papillosa. The levels of IgG1 and IgG2a in mouse sera increased significantly at two weeks after immunization and were highest during the sixth to eighth weeks after immunization. The IgG1 level was higher than IgG2a at all periods of immunization, implicating the dominance of the Th2 response. The levels of IgG1 and IgG2a in the immune sera were shown to be strongly correlated with the numbers of worm recovery, and the correlation coefficient was higher for IgG1. The levels of serum aspartate aminotransferase and alanine transaminase were significantly lower in the sera of rproFgCatL1H-vaccinated mice than in the infected control mice indicating a lower degree of liver damage. This study demonstrated a high potential of FgCatL1H vaccine, and its
Zander, C D; Reimer, L W; Barz, K; Dietel, G; Strohbach, U
Metazoan parasites of guilds of benthic snails and crustaceans and of four fish families--Gobiidae, Gasterosteidae, Syngnathidae, and Zoarcidae--were investigated off the brackish Salzhaff area (Southwest Baltic) in the semienclosed Salzhaff and the near Rerik Riff in the free Baltic. Comparisons revealed greater similarities in parasite populations and communities within the fish guilds than between them. According to an evaluation of the core-/satellite-species concept using abundance values, the most important parasites of fish were some generalists, such as Cryptocotyle spp., Podocotyle atomon, and Diplostomum spathaceum, as well as some specialists, such as Acanthostomum balthicum, Thersitina gasterostei, and Aphalloides timmi. These specialists revealed high degrees of prevalence in their main hosts and lower degrees in one or two by-hosts. Additional importance is assigned to parasites that cause harm to their hosts due to their large size, e.g., Schistocephalus spp., or via massive infestation, e.g., several digenean metacercariae. Because specialists were more prominent in snails and fish from the Rerik Riff, the correlation of host numbers with prevalence resulted in only a slight increase instead of a more rapid rise in regression among crustaceans and fish from the entire Salzhaff, where the generalists were more prevalent. The selected host guilds demonstrated the entire life cycles of three digeneans (P. atomon, A. balthicum, A. timmi), one acanthocephalan (Echinorhynchus gadi), and one nematode (Hysterothylacium sp.). The prevalence increased in these cycles from host level to host level and attained relatively high values in all guilds. The parasite fauna of the Salzhaff area is influenced by eutrophication stress, which leads to a high level of productivity and, consequently, to great densities in primary consumers such as snails and crustaceans. These are attractive for several secondary consumers such as fish and birds, which is the reason for
Full Text Available Abstract Background Paragonimiasis is a neglected tropical disease caused by an infection with lung flukes that is transmitted through the consumption of undercooked crabs. The disease is often confused with tuberculosis. Paragonimiasis is thought to be endemic in south-western Côte d'Ivoire. Methods Two cross-sectional surveys were carried out in the first half of 2009 in patients attending two tuberculosis centres of Abidjan. A third cross-sectional survey was conducted in May 2010 in children of two primary schools in Dabou, where crabs are frequently consumed. Patients with chronic cough provided three sputum samples plus one stool sample. Sputum samples were examined for tuberculosis with an auramine staining technique and for Paragonimus eggs using a concentration technique. Stool samples were subjected to the Ritchie technique. Schoolchildren provided a single stool sample, and samples were subjected to the Kato-Katz and an ether-concentration technique. A pre-tested questionnaire was administered to patients and schoolchildren to investigate food consumption habits. Additionally, between June 2009 and August 2010, shellfish were purchased from markets in Abidjan and Dabou and examined for metacercariae. Results No human case of paragonimiasis was diagnosed. However, trematode infections were seen in 32 of the 272 shellfish examined (11.8%. Questionnaire results revealed that crab and pig meat is well cooked before consumption. Among the 278 patients with complete data records, 62 had tuberculosis, with a higher prevalence in males than females (28.8% vs. 13.9%, χ2 = 8.79, p = 0.003. The prevalence of helminths and intestinal protozoa was 4.6% and 16.9%, respectively. In the school survey, among 166 children with complete data records, the prevalence of helminths and intestinal protozoa was 22.3% and 48.8%, respectively. Boys had significantly higher prevalences of helminths and intestinal protozoa than girls. Hookworm was the
Full Text Available BACKGROUND: The human liver fluke, Opisthorchis viverrini, is designated as a group 1 carcinogen, and is the major risk factor for cholangiocarcinoma in endemic countries throughout Southeast Asia. Proteins in the excretory-secretory products and tegumental surface membranes of the fluke have been proposed to play pivotal roles in parasite survival in the host, and subsequent pathogenesis. These macromolecules are therefore valid targets for the development of vaccines and new drugs to control the infection. Tetraspanins (TSP are prominent components of the tegument of blood flukes where they are essential for tegument formation, are directly exposed to the immune system, and are major targets for a schistosomiasis vaccine. We propose that similar molecules in the surface membranes of O. viverrini are integral to tegument biogenesis and will be efficacious vaccine antigens. METHODOLOGY/PRINCIPAL FINDINGS: The cDNA sequence encoding O. viverrini tetraspanin-1 (Ov-TSP-1 was identified and cloned. The Ov-tsp-1gene was isolated from a cDNA library. Ov-tsp-1 mRNA was expressed most highly in metacercariae and eggs, and to a lesser extent in juvenile and adult worms. Immunolocalization with adult flukes confirmed that Ov-TSP-1 was expressed in the tegument and eggs in utero. Western blot analysis of rOv-TSP-1 probed with sera from O. viverrini-infected humans and hamsters indicated that both hosts raise antibody responses against the native TSP. Using RNA interference we silenced the expression level of Ov-tsp-1 mRNA in adult flukes by up to 72% by 10 days after delivery of dsRNA. Ultrastructural morphology of adult worms treated with Ov-tsp-1 dsRNA displayed a distinctly vacuolated and thinner tegument compared with controls. CONCLUSIONS/SIGNIFICANCE: This is the first report of a tetraspanin from the tegument of a liver fluke. Our data imply that tetraspanins play important structural roles in the development of the tegument in the adult fluke
Zawistowska-Deniziak, A; Wasyl, K; Norbury, L J; Wesołowska, A; Bień, J; Grodzik, M; Wiśniewski, M; Bąska, P; Wędrychowicz, H
Fasciola hepatica infections cause significant global problems in veterinary and human medicine, including causing huge losses in cattle and sheep production. F. hepatica host infection is a multistage process and flukes express papain-like cysteine proteases, termed cathepsins, which play pivotal roles in virulence through host entry, tissue migration and immune evasion. Expression of these proteases is developmentally regulated. Recent studies indicate that excystment of infective larvae is dependent on cysteine proteases and together FhCL3 and FhCB account for over 80% of total protease activity detectable in newly excysted juvenile (NEJ) fluke. This paper focuses on members of the cathepsin L gene family, specifically those belonging to the CL3 clade. The cDNA of two novel cathepsin L3 proteases--FhCL3-1 and FhCL3-2 were cloned. The mRNA transcript expression levels for these enzymes were significantly different at various time points in life development stages obtained in vitro, from dormant metacercariae to NEJ 24h after excystment. Maximum expression levels were observed in NEJ immediately after excystment. In all stages examined by Real Time PCR, FhCL3-2 was expressed at a higher level compared to FhCL3-1 which was expressed only at very low levels. Western blot and immunohistochemical analysis also indicated higher expression of the FhCL3-2 allele and its secretory nature. The ability of antibody responses from rats and sheep challenged with F. hepatica to recognize recombinant FhCL3-1 and FhCL3-2 was shown to differ. Differences were also confirmed through the use of anti-rFhCL3-1 and anti-rFhCL3-2 sera in Western blot analysis of juvenile excretory/secretory (ES) material separated by 2D electrophoresis. These results indicate analysis of relative expression of parasite virulence factors from different populations is required, as this will likely impact the effectiveness of vaccines based on these antigens. Copyright © 2013 Elsevier B.V. All rights
Variação sazonal dos metazoários parasitos de Geophagus brasiliensis (Perciformes: Cichlidae no rio Guandu, Estado do Rio de Janeiro, Brasil - doi: 10.4025/actascibiolsci.v32i2.4883 Seasonal variation of metazoan parasites of Geophagus brasiliensis (Perciformes: Cichlidae from the Guandu river, State of Rio de Janeiro, Brazil - doi: 10.4025/actascibiolsci.v32i2.4883
Jose Luis Luque
Full Text Available O presente trabalho estuda a variação sazonal dos metazoários parasitos de Geophagus brasiliensis do rio Guandu, Estado do Rio de Janeiro, Brasil. Para isso foram realizadas quatro coletas trimestrais de 50 espécimes de G. brasiliensis entre abril de 2006 e março de 2007, nas diferentes estações do ano. Foram coletadas 14 espécies de metazoários parasitos: seis digenéticos (Crassicutis sp. em estágio adulto, e cinco tipos de metacercárias, Austrodiplostomum compactum, Diplostomulum sp., Neascus tipo 1, Neascus tipo 2 e Posthodiplostomum sp.; três nematóides (larvas de Contracaecum sp., adultos de Cucullanus sp. e Procamallanus (Procamallanus peraccuratus ; o monogenético (Sciadicleithrum guanduensis; o acantocéfalo (Neoechinorhynchus paraguayensis; larvas gloquídias de molusco e espécies de hirudíneos das famílias Glossiphonidae e Piscicolidae. Com exceção dos hirudíneos glossifonídeos, todas as espécies apresentaram picos de prevalência e abundância principalmente naqueles peixes coletados na primavera, o que pode indicar uma sazonalidade dessas espécies de parasitos em G. brasiliensis do rio Guandu.The present work studied the seasonal variation of the metazoan parasites of Geophagus brasiliensis from the Guandu river, State of Rio de Janeiro, Brazil. Four collections of 50 specimens of G. brasiliensis were accomplished between April 2006 and March 2007, in the different seasons. A total of 14 species of metazoan parasites were obtained: six trematodes (Crassicutis sp. in adult stage and five metacercariae, Austrodiplostomum compactum, Diplostomulum sp., Neascus type 1, Neascus type 2 and Posthodiplostomum sp., three nematodes (Contracaecum sp. larval, Cucullanus sp. and Procamallanus (Procamallanus peraccuratus adult; monogenean (Sciadicleithrum guanduensis; acanthocephalan (Neoechinorhynchus paraguayensis, larval glochidia; and hirudinean species of Glossiphonidae and Piscicolidae. With the exception of
Variação sazonal dos metazoários parasitos de Geophagus brasiliensis (Perciformes: Cichlidae no rio Guandu, Estado do Rio de Janeiro, Brasil = Seasonal variation of metazoan parasites of Geophagus brasiliensis (Perciformes: Cichlidae from the Guandu river, State of Rio de Janeiro, Brazil
Adriano Reder de Carvalho
Full Text Available O presente trabalho estuda a variação sazonal dos metazoários parasitos de Geophagus brasiliensis do rio Guandu, Estado do Rio de Janeiro, Brasil. Para isso foram realizadas quatro coletas trimestrais de 50 espécimes de G. brasiliensis entre abril de 2006 emarço de 2007, nas diferentes estações do ano. Foram coletadas 14 espécies de metazoários parasitos: seis digenéticos (Crassicutis sp. em estágio adulto, e cinco tipos de metacercárias, Austrodiplostomum compactum, Diplostomulum sp., Neascus tipo 1, Neascus tipo 2 ePosthodiplostomum sp.; três nematóides (larvas de Contracaecum sp., adultos de Cucullanus sp. e Procamallanus (Procamallanus peraccuratus; o monogenético (Sciadicleithrum guanduensis; oacantocéfalo (Neoechinorhynchus paraguayensis; larvas gloquídias de molusco e espécies de hirudíneos das famílias Glossiphonidae e Piscicolidae. Com exceção dos hirudíneos glossifonídeos, todas as espécies apresentaram picos de prevalência e abundânciaprincipalmente naqueles peixes coletados na primavera, o que pode indicar uma sazonalidade dessas espécies de parasitos em G. brasiliensis do rio Guandu.The present work studied the seasonal variation of the metazoan parasites of Geophagus brasiliensis from the Guandu river, State of Rio de Janeiro, Brazil. Four collections of 50 specimens of G. brasiliensis were accomplished between April 2006 and March 2007, in the different seasons. A total of 14 species of metazoan parasites were obtained: six trematodes (Crassicutis sp. in adult stage and five metacercariae, Austrodiplostomum compactum, Diplostomulum sp., Neascus type 1, Neascus type 2 and Posthodiplostomum sp., three nematodes (Contracaecum sp. larval, Cucullanus sp. and Procamallanus (Procamallanus peraccuratus adult; monogenean (Sciadicleithrum guanduensis; acanthocephalan (Neoechinorhynchus paraguayensis,larval glochidia; and hirudinean species of Glossiphonidae and Piscicolidae. With the exception of Glossiphonidae
Zhang, Xiao-Ping; Jiang, Shou-Fu; Hong, Guo-Bao; Fu, Ying-Hua; He, Yan-Yan; Ma, Xiao-Jiang; Wang, Zhen-Yu; Zhang, Yao-Guang; Zhu, Qian; Ma, Xing-Bao
To understand the contamination status of food with parasites in Shanghai market, so as to provide the evidence for formulating the surveillance strategy in parasitic diseases and provide the technical support in the food safety. The freshwater fish products, marine products, vegetables, snails and frogs were sampled with the cluster random method in the farmer-trades and supermarkets of the 18 districts in Shanghai City during 2005-2010 period, and all the specimen were screened by the digestion method, or crushing method or dissection method or saline floatation method to check the parasite metacercaria or larvae or eggs. A total of 5 185 specimen in 23 species of fishes were screened in fresh-water products, with parasite infection rate of 1.93%. About 4 033 specimens of 20 species of fishes and shrimps were screened and 1.76% of samples were infected with Clonorchis sinensis. Among all kind of fishes, the highest infection rate was 7.83% (48/613) in Pseudorasbora parve. No any infected specimen was found among 1 152 specimen of fresh water crustacean screened. Anisakis spp. were found in 12.7% of 433 specimens of 23 species of seawater products, among them, the higher infection rate of Anisakis spp. was found in Pneumatophorus japonicas and Trichiurus haumela with their infection rates of 50.00% (13/26) and 23.46% (42/179), respectively, which much higher than those found in other seawater products (P vegetables, the parasite eggs were found in one of 428 specimens with its infection rate of 0.47%, while no any parasite egg was found in 103 specimens of 10 kinds of fruits. No any Angiostrongylus cantonensis larvae were found in 330 snails, 31.37% of 102 frogs were found infected with Spirometra mansoni spargana. No any contamination with parasites was found in 116 meat specimens of pigs and cattle. In the same time, the intestinal parasite infection rate of residents was 0.42% (131/31 239). It is found that some of foods in Shanghai markets are contaminated
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
Hanna, R E B; Cromie, L; Taylor, S M; Couper, A
Sixteen calves approximately 6 months old were each infected with 500 metacercariae of Fasciola hepatica. Thirty-two days later they were weighed and divided into two groups, and on day 35 all calves in one of the groups were injected subcutaneously with an ivermectin/closantel combination. Both groups were sacrificed between days 70 and 72 to enable counting and examination of the flukes recovered from the bile ducts. Eggs released by the flukes were collected for incubation, hatching and estimation of egg viability. Flukes were counted, flat-fixed in 70% ethanol, stained with catechol and carmine and measured. The reproductive organs, namely testis, vitelline glands, ovary and uterus, were examined and scored on a 0-3 scale according to their state of development. This was visually assessed on the basis of size, distribution and staining density of their constituent tissues and the abundance of eggs in the uterus. A representative sample of flukes from each animal was fixed in formalin for histological sectioning to enable more detailed examination of the reproductive structures. Treatment of the immature flukes reduced the population in cattle by 42.6% as compared with the controls and as a result of the stunting effect due to the presence of closantel during early development the size of treated flukes was reduced by 43.9%. A bimodal pattern of size and reproductive score was also observed in flukes from treated cattle, suggesting that the stunting effect on individual flukes differed depending on whether or not they had gained access to the bile ducts or were still migrating in the hepatic parenchymal tissue at the time of drug exposure with the effect being greater once the fluke had gained access to the bile ducts. The mean reproductive score for untreated flukes was 8.76 and for treated flukes 5.64, a 35.6% reduction. This difference was highly significant (p<0.001). Egg shedding from treated flukes was significantly less than that from controls (p<0
Yang, Qing-Li; Shen, Ji-Qing; Jiang, Zhi-Hua; Shi, Yun-Liang; Wan, Xiao-Ling; Yang, Yi-Chao
Although the responses of inducible nitric oxide synthase (iNOS) and associated cytokine after Clonorchis sinensis infection have been studied recently, their mechanisms remain incompletely understood. In this study, we investigated the effects of toll-like receptor 2 (TLR2) signals on iNOS/nitric oxide (NO) responses after C. sinensis infection. We also evaluated the correlations between iNOS responses and worm development, which are possibly regulated by TLR2 signal. TLR2 wild-type and mutant C57BL/6 J mice were infected with 60 C. sinensis metacercariae, and the samples were collected at 30, 60, 90 and 120 days post-infection (dpi). The total serum NO levels were detected using Griess reagent after nitrate was reduced to nitrite. Hepatic tissue samples from the infected mice were sliced and stained with hematoxylin and eosin (HE) to observe worm development in the intrahepatic bile ducts. The iNOS mRNA transcripts in the splenocytes were examined by real time reverse transcriptase polymerase chain reaction (qRT-PCR), and iNOS expression was detected by immunohistochemistry. Developing C. sinensis juvenile worms were more abundant in the intrahepatic bile ducts of TLR2 mutant mice than those of TLR2 wild-type mice. However, no eggs were found in the faeces of both mice samples. The serum levels of total NO significantly increased in TLR2 mutant mice infected with C. sinensis at 30 (t (5) = 2.595, P = 0.049), 60 (t (5) = 7.838, P = 0.001) and 90 dpi (t (5) = 3.032, P = 0.029). Meanwhile, no changes occurred in TLR2 wild-type mice compared with uninfected controls during the experiment. The iNOS expression in splenocytes showed unexpected higher background levels in TLR2 mutant mice than those in TLR2 wild-type mice. Furthermore, the iNOS mRNA transcripts in splenocytes were significantly increased in the TLR2 wild-type mice infected with C. sinensis at 30 (t (5) = 5.139, P = 0.004), 60 (t (5) = 6.138, P = 0.002) and 90 dpi (t (5) = 6
Cameron, Timothy C; Cooke, Ira; Faou, Pierre; Toet, Hayley; Piedrafita, David; Young, Neil; Rathinasamy, Vignesh; Beddoe, Travis; Anderson, Glenn; Dempster, Robert; Spithill, Terry W
4 IgG is either recognising individual proteins released from extracellular vesicles or is immunoprecipitating intact exosome-like extracellular vesicles. Five extracellular vesicle membrane proteins were identified including two proteins predicted to be associated with vesicle transport/ exocytosis (VPS4, vacuolar protein sorting-associated protein 4b and the Niemann-Pick C1 protein). RNAseq analysis of the developmental transcription of the 38 immunosloughate proteins showed that the sequences are expressed over a wide abundance range with 21/38 transcripts expressed at a relatively high level from metacercariae to the adult life cycle stage. A notable feature of the immunosloughates was the absence of cytosolic proteins which have been reported to be secreted markers for damage to adult flukes incubated in vitro, suggesting that the proteins observed are not inadvertent contaminants leaking from damaged flukes ex vivo. The identification of tegument protein antigens shared between F. gigantica and F. hepatica is beneficial in terms of the possible development of a dual purpose vaccine effective against both fluke species. Copyright © 2017 Australian Society for Parasitology. Published by Elsevier Ltd. All rights reserved.
Full Text Available Madatapa Lake is an oligotrophic lake located in Javakheti plateau (Southern Georgia at 2108 m a.s.l. and is the part of the Javakheti Protected Areas since 2011. The lake have a natural volcanic origin with a surface area of 8.8 km2 and with a maximum depth of 1.7 m. The lake is historically known to be free of fish species which is believed to be a result of severe winter conditions. After the middle 90th the gibel carp [Carassius gibelio (Bloch, 1782] was firstly appeared there. However the exact date and the source of invasion of gibel carp in the lake is unknown and currently untraceable. While gibel carp is a very successful invasive fish species in Georgian inland waters, very limited data is available on its biology, ecology and even distribution as well as its ecological impacts and economic importance in the region. For the first time, we studied the population characteristics of gibel carp in Lake Madatapa (South Georgia with the aim to reveal the extent of its morphological variability, sex ratio, length-weight relationship and its parasitic community. One hundred and forty one individuals presented a mean weight 37.5 g (SD=53, range [13.3-471.7] and average length of 12.5 cm (SD=2.9, range[9.4-30]. The number of lateral line scales and gill rakers varied between 29-31, and 36-47, respectively. Sex was determined in 105 specimens (the gonads of remaining specimens were either strongly damaged by the parasites or were not developed and the sex ratio was significantly female biased (1/1.8, Chi-square test p0.05 between the regression coefficients for males and females and the slope (b do not differ significantly from 3. A sub-sample of thirty four individuals were examined for parasitic infections and 91% (31 were infected with one or two parasites. The plerocercoids of cestoda (Ligula intestinalis L., 1758 had highest prevalence (75%, whereas only 38% of studied fishes were infected with metacercariae of digenean trematode
Donald Arguedas Cortés
Full Text Available Centrocestus formosanus es un parásito trematodo zoonótico originario de Asia asociado con muertes de peces principalmente de cultivo. 907 moluscos provenientes de estanques sembrados con tilapias, seleccionados uno por provincia fueron identificados al nivel taxonómico especifico. Se identificaron cuatro gastrópodos y un bivalvo: M. tuberculata, M. turricula, P. flagellata, H. cubensis y A. luteola. Se reporta, por primera vez, la presencia de dos especies de moluscos en Costa Rica. Se identificaron siete morfotipos de cercarias parasitando las cinco especies de moluscos encontradas. En la segunda exposición experimental se demostró que el morfotipo parapleurolofocercus encontrado en M. tuberculata concuerda con el hallazgo de C. formosanus en alevines de tilapia, después del examen clínico, anatomopatológico y parasitológico realizado a los alevines expuestos. Las metacercarias fueron extraídas del quiste utilizando microagujas y micropinzas lavadas en solución salina fisiológica (0.65%, fijadas en formol caliente al 4% y después esquematizadas con una cámara clara adaptada a un microscopio fotónico, estimándose una abundancia e intensidad media de 1018-1027 digeneos por branquia en cada pez parasitado, determinándose así el hospedador intermediario primario y secundario del parásito. En el presente trabajo se reporta por primera vez Centrocestus formosanus en Costa Rica.Centrocestus formosanus (Opisthorchiida: Heterophyidae as a cause of death in gray tilapia fry Oreochromis niloticus (Perciforme: Cichlidae in the dry Pacific of Costa Rica. Centrocestus formosanus is a zoonotic trematode from Asia and has been mainly associated as cause of death of cultured fish. To identify pathogen trematode species in tilapia fry (Oreochromis niloticus and to determine mollusks hosting these parasites, freshwater mollusks were collected from tilapia cultured ponds and experimental infections were carried out with tilapia fries and