The occurrence of metacercariae of Bucephaloides gracilescens (Digenea: Gasterostomata) in an intermediate host, the four-bearded rockling, Enchelyopus cimbrius (Gadidae)

Infections with metacercariae of Bucephaloides gracilescens in the four-bearded rockling Enchelyopus cimbrius were studied for a year in a Norwegian fjord. The prevalence was at all times high, but abundance showed a marked decline in spring to a low level during summer. A rise in abundance, variance-to-mean ratio and frequency of smaller metacercariae in early autumn (August-September) shows infection to have occurred mainly in autumn. There were no differences in level of infection between host sexes, but a highly significant positive correlation of parasite burden with host length. No influence on the condition factor of the host was revealed, the parasite cysts in the central nervous system are believed to have low pathogenicity. A detailed account of the distribution of metacercarial cysts is given. No support for infection being via the lateral line canals is revealed, cercarial penetration is believed to be through the general surface of the head and through the unpaired fins of the trunk and tail.     

Two species of goby, Boleophthalmus pectinirostris and Scartelaos sp., as the new second intermediate hosts of heterophyid fluke in Korea

A survey was performed in order to determine the infection status of the metacercariae of heterophyid fluke in two goby species, Boleophthalmus pectinirostris and Scartelaos sp., collected from Gangjin-gun, and Shinangun, Sooncheon-shi, Jeollanam-do, Republic of Korea. A total of three metacercariae of Heterophyopsis continua was found in only one B. pectinirostris (10.0%) from Gangjin-gun. Heterophyes nocens metacercariae were detected in 24 B. pectinirostris (96.0%) and 14 Scartelaos sp. (63.6%) from Shinan-gun. Heterophyopsis continua metacercariae were found in 11 B. pectinirostris (44.0%) and 21 Scartelaos sp. (95.5%) from Shinan-gun. Stictodora fuscata metacercariae were detected in 18 B. pectinirostris (72.0%) from Shinan-gun. No metacercariae were detected in 20 B. pectinirostris from Sooncheon-shi. From the above results, this study is the first to prove that B. pectinirostris and Scartelaos sp. serve as the second intermediate hosts of some heterophyid flukes in Korea.     

An ultrastructural study of the cercarial excretory system in Bucephaloides gracilescens and Prosorhynchus squamatus

The ultrastructure of the flame cells, capillaries, collecting tubes, excretory bladder, excretory atrium, caudal vesicle, lateral caudal ducts and excretory pores of cercariae of Bucephaloides gracilescens (Rudolphi, 1819) Hopkins, 1954 and Prosorhynchus squamatus Odhner, 1905 (Digenea: Bucephalidae) is described. Both species are essentially similar except for some details. The terminal parts of the protonephridia have all the structural features that are typical of trematodes. The collecting tubes in the cercarial body are composed of cells that are wrapped around the lumen. The main collecting tubes are joined to the excretory bladder syncytium by septate junctions. Features of P. squamatus excretory bladder epithelium indicate that it is involved in secretory activity, but this is not the case in B. gracilescens. In both species the luminal surface of the excretory bladder epithelium is increased by lamellae, and the basal plasma membrane forms invaginations. In the bladder syncytium of P. squamatus both apical lamellae and basal invaginations are more developed and mitochondria are also more numerous. The excretory atrium is lined by a syncytium with nucleated cytons located in the surrounding parenchyma. The atrium lining is not continuous with the body tegument and possesses specific secretory inclusions and a thick glycocalyx. Septate junctions connect the atrium syncytium to the excretory bladder epithelium at its anterior end and to the syncytial excretory epithelium lining the caudal vesicle and the lateral caudal ducts at its posterior. In the excretory pores the caudal duct syncytium is joined to the tegument by septate desmosomes.     

Prosorhynchoides gracilescens (Rudolphi, 1819) (Digenea: Bucephalidae) from Lophius piscatorius L. is a species complex: a redescription of this species (sensu stricto) from the western Mediterranean and the description of P. borealis n. sp. from the northern North-East Atlantic

The bucephalid digenean Prosorhynchoides gracilescens (Rudolphi, 1819) is a common intestinal parasite of the angler fish Lophius piscatorius in European marine waters. Detailed studies of new material collected off the coasts of both Marseilles and Corsica in the western Mediterranean, and comparison with museum material from the northern North-East Atlantic, indicated that the accepted concept of this species comprises two distinct taxa. The Mediterranean form occurs in relatively small numbers and has small eggs, a large rhynchus and a vitelline distribution that finishes well short of the rhynchus, whereas the NE Atlantic form, which comprises the majority of records, occurs often in large numbers, has larger eggs, a relatively smaller rhynchus and a vitelline distribution that tends to reach the rhynchus. Since the type-material is from the Mediterranean and resembles the Mediterranean form, there was little option but to consider the latter as P. gracilescens (sensu stricto) and to describe the NE Atlantic form as a new species, for which the name P. borealis n. sp. is coined to reflect its northerly distribution. Both species are described and figured in detail, and a table and illustration of the diagnostic features are presented. In addition, some discussion is included on the distribution of these two species and whether P. gracilescens (s. str.) might be a relict species, and on non-European records of P. gracilescens (sensu lato).     

Surveys of Gynaecotyla squatarolae and Microphallus koreana (Digenea: Microphallidae) Metacercariae in Two Species of Estuarine Crabs in Western Coastal Areas, Korea

To figure out the geographical distribution of Gynaecotyla squatarolae and Microphallus koreana metacercariae in Korea, shore crabs of southwestern coastal areas were examined. Eight coastal areas in Inchon-si (A), Gyeonggi-do (B), Chungcheongnam-do (C, D, and E), Jeollabuk-do (F), and Jeollanam-do (G and H) were selected, and 2 kinds of crabs, Macrophthalmus dilatatus and/or Macrophthalmus japonicus, were caught. After transportation to the laboratory, 15 crabs per each group were grouped and ground in a mortar and pestle, and examined for microphallid metacercariae. In M. dilatatus, G. squatarolae metacercariae were recovered from 3 (C, E, and H) out of 6 regions, but M. koreana metacercariae were not recovered. In the case of M. japonicus, G. squatarolae metacercariae were recovered from 6 (B, D, E, F, G, and H) of 7 areas surveyed, and M. koreana matacercariae were detected from 5 regions (A, B, D, F, and H). These results indicate that the life cycle of G. squatarolae is maintained in the western coastal areas using M. dilatatus and M. japonicus as intermediate hosts, while that of M. koreana is maintained only using M. japonicus.     

c-fos Expression as a Model for Studying the Action of Hexachlorocyclohexane Isomers in the CNS

The induction of protooncogene c-fos in the CNS after administration of several convulsants has been studied. The organochlorine insecticide gamma-hexachlorocyclohexane (gamma-HCH, lindane) has been shown to induce c-fos expression in different brain areas. Pentylenetetrazole and picrotoxin, a known gamma-aminobutyric acid-receptor antagonist, have also been considered. The administration of two nonconvulsant isomers of gamma-HCH, alpha-HCH, and delta-HCH, before the mentioned toxicants, affects the protooncogene expression in different ways. The differential pattern of expression displayed by c-fos after these treatments suggests the presence of diverse mechanisms of action for the compounds studied.     

Pseudomonas sp. to Sphingobium indicum: a journey of microbial degradation and bioremediation of Hexachlorocyclohexane

The unusual process of production of hexachlorocyclohexane (HCH) and extensive use of technical HCH and lindane has created a very serious problem of HCH contamination. While the use of technical HCH and lindane has been banned all over the world, India still continues producing lindane. Bacteria, especially Sphingomonads have been isolated that can degrade HCH isomers. Among all the bacterial strains isolated so far, Sphingobium indicum B90A that was isolated from HCH treated rhizosphere soil appears to have a better potential for HCH degradation. This conclusion is based on studies on the organization of lin genes and degradation ability of B90A. This strain perhaps can be used for HCH decontamination through bioaugmentation.     

Infection status of sweetfish from Kwangjung-stream and Namdae-stream in Yangyang-gun, Kangwon-do with the metacercariae of Metagonimus yokogawai

This study was performed to observe the infection status of sweetfish, caught from Kwangjung-stream and Namdae-stream in Yangyang-gun, Kangwon-do in August 1989, with the metacercariae of Metagonimus yokogawai. All of 28 sweetfish from Kwangjung-stream were infected with the metacercariae of M. yokogawai. The number of metacercariae in a fish ranged 89-521(224 on average), and the number of larvae/g of flesh was 12 approximately 55 (22 on average). On the other hand, no metacercariae were detected in the fish from Namdae-stream. It has been confirmed that the Kwangjung-stream in Yangyang-gun, Kangwon-do is one of the endemic foci of metagonimiasis in Korea.     

Effects on DHEA levels by estrogen in rat astrocytes and CNS co-cultures via the regulation of CYP7B1-mediated metabolism

The neurosteroid dehydroepiandrosterone (DHEA) is formed locally in the CNS and has been implicated in several processes essential for CNS function, including control of neuronal survival. An important metabolic pathway for DHEA in the CNS involves the steroid hydroxylase CYP7B1. In previous studies, CYP7B1 was identified as a target for estrogen regulation in cells of kidney and liver. In the current study, we examined effects of estrogens on CYP7B1-mediated metabolism of DHEA in primary cultures of rat astrocytes and co-cultures of rat CNS cells. Astrocytes, which interact with neurons in several ways, are important for brain neurosteroidogenesis. We found that estradiol significantly suppressed CYP7B1-mediated DHEA hydroxylation in primary mixed CNS cultures from fetal and newborn rats. Also, CYP7B1-mediated DHEA hydroxylation and CYP7B1 mRNA were markedly suppressed by estrogen in primary cultures of rat astrocytes. Interestingly, diarylpropionitrile, a well-known agonist of estrogen receptor β, also suppressed CYP7B1-mediated hydroxylation of DHEA. Several previous studies have reported neuroprotective effects of estrogens. The current data indicate that one of the mechanisms whereby estrogen can exert protective effects in the CNS may involve increase of the levels of DHEA by suppression of its metabolism.     

Histopathological and ultrastructural observations of metacercarial infections of Diplostomum phoxini (Digenea) in the brain of minnows Phoxinus phoxinus

The spatial distribution and histopathological changes induced by metacercariae of the digenean trematode Diplostomum phoxini (Faust, 1918) in the brains of European minnows Phoxinus phoxinus (L.) from the River Endrick, Scotland, were studied by light and electron microscopy. Post-mortem examination of a sample of 34 minnows revealed that 50% (n = 17) of the population was infected with 13.7 +/- 2.6 (mean +/- SE; range 1 to 38) metacercariae per infected host. Serial histological sections of the infected minnow brains revealed that the metacercariae were unevenly distributed throughout the brain, with aggregations occurring in the cerebellum, the medulla oblongata and the optic lobes. In fish with highest intensities of infection, over 40% of the cerebellar area and about 30% of the medulla oblongata area were occupied by larvae. Metacercariae disrupt the integrity of brain tissue, with individuals being found in small pockets surrounded by cellular debris. Metacercariae were rarely encountered on the surface of the brain. Electron microscopic examination of infection sites revealed that the granular layer surrounding metacercariae was necrotic, exhibited nuclear degradation and was marked by vacuolation of the cytoplasm. Rodlet cells, the only inflammatory cell types recorded in this study, were found only in parasitized brains and in close proximity to the teguments of metacercariae. It is hypothesised that secretions released from the teguments of metacercariae are a counter response to protect the metacercariae from the fish brain's cellular defence mechanisms.     

The effect of pesticides on the development in rats of osteosarcomas induced by 90Sr]

In experiments with rats the authors have assessed the development of remote effects of radiation (90Sr) combined with chronic exposure to chlorophos and lindane within a wide range of doses. It has been shown that chronically administered chlorophos and lindane have no carcinogenic and cocarcinogenic effects.     

Effects of copper sulfate toxicity on cercariae and metacercariae of Echinostoma caproni and Echinostoma trivolvis and on the survival of Biomphalaria glabrata snails

Copper in the form of copper sulfate (CuSO4) decreases the survival of Biomphalaria glabrata snails, but the effects of this molluscicide on Echinostoma caproni and Echinostoma trivolvis, 2 species of digeneans that use B. glabrata as intermediate hosts, are not known. Studies were done on the effects of various concentrations of CuSO4 in artificial spring water (ASW) on the survival and infectivity of E. caproni and E. trivolvis cercariae. Solutions containing 1.0, 0.1, and 0.01% CuSO4 were 100% lethal within 2 hr of exposure for both species. Time to 50% mortality in 0.001% CuSO4 was 8 hr for E. caproni and 16 hr for E. trivolvis; at 24 hr, the controls showed 50 and 65% mortality, respectively. Treatment of cercariae of both species for 0.5 hr in 0.001% CuSO4 had no effect on the ability of cercariae to form normal cysts in juvenile B. glabrata snails. However, treatment with 0.01% CuSO4 for 0.5 hr caused a significant reduction in the ability of cercariae of both species to encyst in snails. Treatment of encysted metacercariae of both species in 0.001% CuSO4 for I hr had no effect on subsequent excystation of these echinostomes in a trypsin-bile salts medium, whereas concentrations of 1.0, 0.1, and 0.01% CuSO4 and 1.0 and 0.1% CuSO4 decreased chemical excystation of E. caproni and E. trivolvis cysts, respectively. Survival studies on the effects of CuSO4 in Locke's solution on chemically excysted metacercariae of both species were also done. Excysted metacercariae of both species were killed by 2 hr in either 0.1 or 0.01% CuSO4 in Locke's solution. However, time to 50% mortality for both species of excysted metacercariae in 0.001% CuSO4 was approximately 5 hr. Time to 50% mortality for the controls was about 12 hr. Survival of juvenile B. glabrata snails was also examined. All B. glabrata snails were dead by 6 hr in 1 and 0.1% CuSO4 in ASW. Biomphalaria glabrata snails showed 50% mortality by about 6 hr in 0.01% CuSO4 and about 80% were still alive at 24 hr in 0.001% CuSO4. All controls were alive at 24 hr, at which time the experiment was terminated. Concentrations greater than 0.001% CuSO4 increased snail mortality, as well as that of the cercariae and excysted metacercariae of E. caproni and E. trivolvis. Our findings suggest that concentrations of copper sufficient to eliminate juvenile B. glabratta snails are also sufficient to kill the cercariae and excysted metacercariae of these digeneans but not the encysted metacercariae, which may be protected by their cyst walls.     

Studies on the viability of metacercariae of Fasciola gigantica.

The viability of metacercariae of Fasciola gigantica was tested by in vitro and in vivo methods. In vitro testing was based upon the motility of juvenile flukes within the inner cyst as examined under the light microscope. In vivo testing was undertaken through experimental infections of rabbits (two groups) and natural definitive hosts, lambs (one group). In the first group, out of six rabbits each given 25 metacercariae, worm establishment only took place in one rabbit with a single fluke recovery on 60 days post infection. In the second group of six rabbits each given 200 metacercariae, five were infected, with two or three flukes per host. All the lambs given 250 metacercariae became infected showing prevalences of 7.2-40% in comparison with rabbits in which low prevalences (0-4%) were recorded. The results indicated that even viable metacercariae which were already tested in vitro could not readily establish in rabbits. Such variability in worm establishment suggests that immunological and chemotherapeutic studies in rabbits infected with F. gigantica are likely to be unreliable.     

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

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

Effects of lindane on the glucose metabolism in rat brain cortex cells.

The influence of 0.5 mM gamma-hexachlorocyclohexane (gamma-HCH, lindane) on glucose transport has been investigated using the analog 3-O-methyl-D-(U-14C)glucose. The glucose uptake was lineal for at least 10 sec. Preincubation of dissociated brain cortex cells with lindane decreased the transport of glucose with respect to the controls. The treatment of brain cortex cells with other organochlorine compounds indicated that the alpha-, delta-HCH isomers and dieldrin reproduced the same inhibitory pattern, while beta-HCH and endrin were inactive. The total radioactivity incorporated into CO2 from (U-14C) glucose in the cerebral cortex is also inhibited by lindane in a time dependent manner.     

Excystation and development in the chick and on the chick chorioallantois of the metacercaria of Sphaeridiotrema globulus (Trematoda)

Fried B. and Huffman J. E. 1982. Excystation and development in the chick and on the chick chorioallantois of the metacercaria of Sphaeridiotrema globulus (Trematoda). International Journal for Parasitology12: 427–431. Encysted metacercariae of Sphaeridiotrema globulus were obtained from naturally infected Goniobasis virginica snails in Morris County, New Jersey, U.S.A. These metacercariae were successfully excysted within l h in an alkaline bile salts trypsin medium maintained at 41°C in the absence of acid pepsin pretreatment. Encysted metacercariae treated in 3% sodium bicarbonate produced successful infections in day-old domestic chicks and worms became ovigerous in the lower ileum by day 4. Excysted metacercariae transferred to the chorioallantois of 6–10-day-old chick embryos maintained at 41°C developed into ovigerous adults by day 4.     

Alleviation of lindane induced toxicity in testis of Swiss mice (Mus musculus) by combined treatment with vitamin C, vitamin E and alpha-lipoic acid

Mitigation of lindane induced toxicity in testis of Swiss mice by combined treatment with vitamin C, vitamin E and alpha-lipoic acid has been evaluated. Male healthy mice (40), 8-10 weeks old were randomly selected and divided into 4 groups, control (C); lindane (L); antioxidant (A) and antioxidant plus lindane (A+L). Group C animals were administered only the vehicle (olive oil); in group L lindane was administered orally at a dose of 40 mg/kg body wt.; in group A combination of antioxidants at a dose of 125 mg/kg body wt.(vitamin C: 50 mg/kg body wt., vitamin E: 50 mg/kg body wt. and alpha-lipoic acid: 25 mg/kg body wt.) was administered orally; in group A+L both antioxidants (125 mg/kg body wt.) and lindane (40 mg/kg body wt.) were administered at their respective doses. In group A+L antioxidants were administered 1 h prior to lindane administration. All treatments were continuously given for 60 days. Histopathological changes due to lindane intoxication indicated shrunken and distorted seminiferous tubules, sparse Leydig cells and blood vessels and atrophy in the tissue. The testis weight also decreased significantly. Lindane treated group showed increased lipid peroxidation, whereas glutathione, glutathione peroxidase, superoxide dismutase, catalase and protein were significantly decreased compared to control. Lindane induced damage was minimized by administration of antioxidants. Results suggest that combined pretreatment with antioxidants can alleviate the damage caused to testis by lindane.     

Low doses of the pesticide lindane induce protein release by the fat body of female cockroach Blaberus craniifer (Dictyoptera)

In the ovoviviparous cockroach Blaberus craniifer, low doses of the pesticide lindane (1-6 microg/g of body mass) have been implicated in the enhancement of ovarian growth and vitellogenesis onset in headless female ovaries. In order to investigate lindane effects on protein release by the fat body, we used antibodies raised against egg proteins to quantify protein levels in fat body, hemolymph and ovaries of treated-fed or -decapitated females 3- or 5-days -old. In vitro assays used fat body in Grace's medium to quantify the protein amount released in the medium. Individual data for each treatment were related to their corresponding control in paired series. In vivo, ovarian enhanced protein content was linked to an enhanced protein secretion by the fat body. This was ascertained in vitro by high levels of released protein in the medium containing lindane (1 microM) by fat body from females, but not from males. This effect was inhibited by EDTA, a calcium chelator. The present results confirmed that low doses of lindane (about 3 microg/g of body mass) acted as a juvenile hormone analogue, at the level of the ovaries, by enhancing protein uptake, and also at the level of the fat body, by triggering protein release. This property is calcium-dependent.     

Patterns of fish deformities and their association with trematode cysts in the Willamette River,Oregon

Synopsis We described skeletal deformities in Willamette River fishes from larval and juvenile specimens collected in 2002 and 2003. Deformities were found in most taxa examined but were more frequent in native broadcast spawners, especially minnows and suckers, than in native or exotic nest builders. Caudal deformities were uniformly distributed throughout the river, but precaudal deformities were more localized near the towns of Newberg and Salem (Wheatland Ferry). In northern pikeminnow, Ptychocheilus oregonensis, deformities were dependent on relative hatch date, with early season fish having about three times the deformity load as late season fish. In a subsample of northern pikeminnow and chiselmouth, Acrocheilus alutaceus, number of deformities was directly related to number of trematode metacercariae and precaudal deformities were twice as likely as caudal deformities to be associated with metacercariae. Based on a logistic regression, the probability of a precaudal deformity was dependent on number of metacercariae and geographic area with the area effect disappearing as the number of cysts increased. A separate analysis showed that some types of deformities were unlikely to be associated with metacercariae. However, even in cases where metacercariae were unlikely to be associated with deformities, metacercariae were usually present elsewhere in the fish and an indirect effect could not be dismissed. The taxonomic, spatial and temporal patterns of skeletal deformities in Willamette River fishes may be due to differences in intermediate host (snail or fish) resistance or susceptibility to parasites, to differences in life history ecology, or to anthropogenic effects that are manifest in increased precaudal deformities near Newberg and Wheatland Ferry or decreased rates elsewhere.     

Seasonal occurrence of Dollfustrema vaneyi (Digenea: Bucephalidae) metacercariae in the bullhead catfish Pseudobagrus fulvidraco in a reservoir in China

The seasonal population dynamics of metacercariae of the bucephalid Dollfustrema vaneyi (Tseng, 1930) Echmann, 1934 in the bullhead catfish Pseudobagrus fulvidraco (Richardson) were investigated in Jiangkou reservoir, Jiangxi Province, east China, during the period from April 1990 to August 1991. In total, 523 fish were obtained, and the overall prevalence of the metacercariae was 89.87% and mean abundance 136.25 +/- 308.09 (mean +/- SD). A pattern of seasonal changes in prevalence and mean abundance was observed, with higher levels of metacercariae infection in late spring and summer. An analysis of the distribution of D. vaneyi in different organs of P. fulvidraco suggested that the eyes might be a suitable location for the parasite. Furthermore, the possible role of metacercariae in bullhead catfish was discussed in relation to the life cycle of this parasite.