Schistosoma mekongi and Schistosoma japonicum: Differences in the distribution of eggs in the viscera of mice

The difference in the distribution of Schistosoma eggs in the viscera has not been clearly elucidated in the two closely related species Schistosoma japonicum and Schistosoma mekongi. In this study, we quantitatively compared the distribution of eggs in mice infected with the two species. In S. mekongi-infected mice, 56.6% to 69.4% of total eggs were found in the distal small intestine 9 to 15 weeks after infection, while in S. japonicum-infected mice, 48.8% to 71.8% of eggs were found in the proximal small intestine during the same period. There were significantly more eggs in the liver in mice infected with S. japonicum than in those infected with S. mekongi. The number of adult worms recovered did not differ between the two species during the study period. The total number of eggs laid in the tissues also did not differ between the two species at 12 to 15 weeks postinfection, but in the earlier period the total number of eggs was significantly fewer in S. mekongi-infected than in S. japonicum-infected mice, suggesting the delayed maturation of the former compared with the latter. These results clearly show that S. japonicum and S. mekongi exhibit different oviposition behavior in their hosts.     

Schistosoma malayensis n. sp.: a Schistosoma japonicum-complex schistosome from Peninsular Malaysia

Schistosoma malayensis n. sp., a member of the Schistosoma japonicum complex is described from Rattus muelleri in Peninsular Malaysia and 2 strains are characterized. The only morphological differences noted among adults from natural hosts were that S. malayensis are in general smaller than S. mekongi and S. japonicum. But these differences may be the result of host-induced variations and therefore are of little taxonomic value. To minimize the effects of host-induced variations, adult worms recovered from laboratory mice with similar worm burdens at 50-56 days postinfection were compared. These comparisons revealed only minor morphometric differences among these 3 species. Schistosoma malayensis eggs from naturally and experimentally infected hosts are most similar to those of S. mekongi, with eggs of both species being, in general, smaller than those of S. japonicum. The egg index for S. malayensis is usually higher than for S. japonicum and lower than for S. mekongi. Differences were noted in the developmental rates in mice for 2 isolates of S. malayensis, S. mekongi, and S. japonicum (Philippine strain), but relatively large differences observed between isolates of S. malayensis indicate that, in this case, the developmental rate is not a useful taxonomic character. Schistosoma malayensis is erected principally on the basis of differences, reported elsewhere, in the life histories and in the electrophoretic migration patterns of isoenzymes of adult worms as compared to S. mekongi and S. japonicum. These comparisons indicate that S. malayensis is more closely related to S. mekongi than to S. japonicum.     

Experimental Life History and Biological Characteristics of Fasciola gigantica (Digenea: Fasciolidae)

This study was conducted to investigate the life history, morphology, and maturation of larval stages and adult worms of Fasciola gigantica in experimental mice. Lymnaea auricularia rubiginosa was used as the intermediate host, and Oryza sativa was used for encystment of the metacercariae, while Mus musculus was used as the definitive host for maturation study. Fresh eggs from the gall bladder of water buffaloes fully developed into embryonated ones and hatched out at days 11-12 after incubation at about 29ºC. Free-swimming miracidia rapidly penetrated into the snail host, and gradually developed into the next larval stages; sporocyst, redia, and daughter redia with cercariae. Fully-developed cercariae were separated from the redia and shed from the snails on day 39 post-infection (PI). Free-swimming cercariae were immediately allowed to adhere to rice plants, and capsules were constructed to protect metacercariae on rice plants. Juvenile worms were detected in intestines of mice at days 3 and 6 PI, but they were found in the bile duct from day 9 PI. Juvenile and adult flukes were recovered from 16 mice experimentally infected with metacercariae, with the average recovery rate of 35.8%. Sexually mature adult flukes were recovered from day 42 PI. It could be confirmed that experimentally encysted metacercariae could infect and develop to maturity in the experimental host. The present study reports for the first time the complete life history of F. gigantica by an experimental study in Thailand. The obtained information can be used as a guide for prevention, elimination, and treatment of F. gigantica at environment and in other hosts.     

Host-parasite relationship of Angiostrongylus cantonensis. 1. Intracranial transplantation into various hosts.

Various stages of Angiostrongylus cantonensis recovered from the brain of experimentally infected mice were transplanted intracranially into rats. Third and fourth-stage worms recovered 2--7 days postinfection were able to develop normally after transplantation into recipient rats. The fifth-stage worm obtained 14--15 days postinfection would enter the brain tissue of rats but died shortly afterwards. However, the same stage of worms recovered from rats, after a similar transplantation, were found to develop normally in the recipient animal. Young fifth-stage worms, from the subarachnoid space of rats, which were ready for the pulmonary migration were also transplanted into rabbits but the worms failed to reach the lungs. In the control rat-to-rat transplantation, the worms successfully completed the pulmonary migration. The morphogenesis and initial growth rate of A. cantonensis were similar in both mice and rats but in the former host the worms started to grow at a markedly slower rate after the last moult and gradually degenerated.     

Penetration and maturation of Schistosoma mansoni in suckling and adult Swiss Webster and DBA/2 mice

Murine schistosomosis is a widely used experimental model of the human disease. Different methods have been employed to infect mice with Schistosoma mansoni cercariae, such as subcutaneous or intraperitoneal injections, the tail immersion technique, and the use of a metal ring placed on the abdominal skin of anaesthetized animals. An alternative method of infection that requires no anaesthesia and no restraint is to place suckling mice (10 days old) on a Petri dish with a small volume of water containing cercariae. In this study, we compared the penetration and maturation of S. mansoni in mice infected by this alternative method with those noted in mice infected at an older age (45 days) by the tail immersion technique. Besides evaluating the effects of age and method of infection, we also compared the susceptibility of two strains of mice (Swiss Webster (SW) and DBA/2). Mice were exposed to 100 cercariae and worms (by portal-hepatic perfusion) as well as eggs were recovered in the liver and intestines on postinfection (PI) days 35, 55 and 90. Skin penetration was very efficient (about 100%) irrespective of the mouse strain, sex, age and method of infection. Worm and egg recoveries were higher in SW mice at any PI interval, but strain differences tended to be less pronounced on PI day 90. In both strains, recoveries of worms and eggs were clearly higher in mice infected at a younger age (10 days old). This study thus suggests that infection of free-moving suckling mice is a suitable alternative to other methods of infection with S. mansoni.     

东方人类血吸虫及其螺蛳宿主交互感染的研究

以我国安徽及菲律宾莱特岛的日本血吸虫、湄公血吸虫、马来西亚类似日本血吸虫以及中华血吸虫交互感染其螺蛳宿主。结果表明日本血吸虫及钉螺与湄公血吸虫、马来西亚类似日本血吸虫及其螺蛳宿主亲缘关系较疏远。而湄公血吸虫与马来西亚类似日本血吸虫及其螺蛳宿主有较近的亲缘关系。交互感染结果为马来西亚类似日本血吸虫及钙河罗氏螺的分类问题提供了新的依据。     

Recovery,growth, and development of Acanthoparyphium tyosenense (Digenea: Echinostomatidae) in experimental chicks

Chicks were experimentally infected with Acanthoparyphium tyosenense (Digenea: Echinostomatidae) metacercariae per os, and the growth and development of worms in this host were observed from days I to 38 postinfection (PI). The worms grew rapidly and matured sexually in the small intestine (chiefly in the jejunum) of chicks by day 5 PI. and survived at least up to day 38 Pi, although worm recovery decreased after day 5 PI. Both parenchymal and reproductive organs increased greatly in size from day 2 to day 10 PI and then continued to increase gradually in size up to day 38 PI. The number of uterine eggs reached a peak on days 10 and 15 PI and then decreased gradually. The results suggest that chicks are a fairly suitable definitive host for experimental infection with A. tyosenense.     

Longevity of Zygocotyle lunata in Balb/c mice and long-term survival of digeneans in vertebrate hosts

This study used Balb/c mice to examine the longevity of Zygocotyle lunata in a murine host. Of 11 mice, each exposed to 20 Z. lunata cysts, six were infected with a total of 12 worms from 11 to 24 weeks postinfection (PI). Live worms recovered at 24 weeks PI had a mean body area of about 25?mm2. These worms produced viable eggs with well-developed miracidia following embryonation in artificial spring water for 2 weeks at 28°C. The Balb/c mouse is a useful model to study longevity of this paramphistomid trematode for at least 6 months PI. An additional aspect of this article is a review of the pertinent literature published from 1937 to 2007 on ageing and longevity of digeneans.     

Reproductive potential of Echinococcus multilocularis in experimentally infected foxes, dogs, raccoon dogs and cats

A total of 15 red foxes, 15 raccoon dogs, 15 domestic dogs and 15 domestic cats were each infected with 20,000 protoscolices of Echinococcus multilocularis. At 35, 63, and 90 days post inoculation (dpi), five animals from each group were necropsied and the worm burdens determined. The highest worm burdens in foxes (mean of 16,792) and raccoon dogs (mean of 7930) were found at 35 dpi. These declined to a mean of just 331 worms in foxes and 3213 worms in raccoon dogs by day 63 with a further decline to 134 worms in foxes and 67 worms in raccoon dogs by day 90. In dogs, there was no significant difference between worm burdens recovered at days 35 (mean of 2466) and day 90 (mean of 1563), although reduced numbers were recovered on day 63 (mean of 899). In cats, worms were found in four animals 35 dpi (mean of 642), in three at 63 dpi (mean of 28) and in two at 90 dpi (mean of 57). Faecal egg counts were determined at 3 day intervals from 25 dpi. A mathematical model of egg excretion dynamics suggested that the mean biotic potential per infected animal was high in foxes (346,473 eggs); raccoon dogs (335,361 eggs) and dogs (279,910 eggs) but very low for cats (573 eggs). It also indicated that approximately 114, 42 and 27 eggs per worm were excreted in the faeces of dogs, raccoon dogs and foxes, respectively. The fecundity of worms in cats was low with an average of less than one egg per worm. The peak levels of coproantigen were detected earlier in foxes and raccoon dogs than in dogs. Eggs recovered from foxes, raccoon dogs and dogs resulted in massive infections in experimental mice. However, metacestodes did not develop from eggs originating from infected cats. It is concluded that foxes, raccoon dogs and dogs are good hosts of E. multilocularis. In contrast, the low worm establishment, the very few excreted eggs and the lack of infectivity of eggs strongly indicate that cats play an insignificant role in parasite transmission.     

Effect of Schistosoma mansoni infection on fecundity and perivitelline fluid composition in Biomphalaria glabrata

Egg production in the snail, Biomphalaria glabrata, infected with Schistosoma mansoni declined on day 23 postinfection, and was significantly lower than uninfected control snails by day 28 and thereafter. Protein and galactogen content of eggs produced by infected snails did not change during the period of reduced fecundity. This suggests that decreased hemolymph nutrient levels (rather than depleted albumen gland reserves) are responsible for inhibition of snail egg production. Growth rates of infected and uninfected snails were indistinguishable from days 14 through 35 postinfection. The hatching success of eggs produced by infected snails decreased slightly beginning at day 21 postinfection.     

Dynamic analysis of T-lymphocyte function in relation to hepatopathologic changes and effect of interleukin-12 treatment in mice infected with Schistosoma japonicum

We analyzed the dynamics of splenic T-lymphocyte function in relation to hepatopathologic changes in C3H/Hc mice, experimentally infected with Schistosoma japonicum. Vigorous granuloma formation was observed at 7 wk postinfection. At 10 wk postinfection, granuloma formation entered into the down-modulation stage, as represented by the diminished granuloma size. The Th2 response was activated when eggs appeared in the liver, whereas Th1 responses were depressed and the proliferation of T lymphocytes was decreased. The level of IgG antibodies to the worm and egg antigens rose continually after infection. Interleukin-12 treatment of infected mice inhibited Th2 responses and T-cell proliferation, decreased granuloma formation and fibrosis, but had no effect on the fecundity of the worms. These data suggest that egg deposition is the major factor driving Th2 responses, depressing Th1 cytokine expression as well as T-cell proliferation in S. japonicum-infected mice.     

Oxamniquine, praziquantel and lovastatin association in the experimental Schistosomiasis mansoni

The activity of lovastatin associated with oxamniquine or praziquantel against schistosomiasis mansoni was evaluated in mice infected with Schistosoma mansoni. Forty days after infection, mice were treated with lovastatin, 400 mg/kg for five consecutive days by oral route, and on the last day of this sequence with 50 mg/kg oxamniquine or with 200 mg/kg praziquantel, both by oral route, single dose. Fifteen days later, the animals were perfused in parallel with an untreated control group. Studies were carried out in vitro, using lovastatin in culture medium containing S. mansoni worms proceeding from experimentally infected mice. In the in vivo trials, the association of lovastatin with oxamniquine or praziquantel did not show any additive action, but there were oogram changes when lovastatin was associated with oxamniquine. In vitro lovastatin was able to interrupt the maturation of S. mansoni eggs, which remained at the 1st or 2nd stages, depending on the dose used. The total number of morphologically dead eggs found in culture of worms exposed to 2 microg/ml or 4 microg/ml concentrations of lovastatin was significantly higher than the number of viable eggs. Using the probe Hoescht 33258 it was observed that 70% of the eggs considered morphologically viable in the treated groups (against 16% in the control group) were labeled, indicating that the majority of the viable eggs had membrane permeability increased due to lovastatin action.     

Cultivation of excysted metacercariae of Echinostoma caproni (Trematoda) to ovigerous adults on the chick chorioallantois

Excysted metacercariae of Echinostoma caproni were cultivated on the chick chorioallantoic membrane (CAM) maintained at 38.5 +/- 1 C and a relative humidity of 60-65%. Of 59 6-day-old embryos, each inoculated with 25 metacercariae, 29 (49.2%) were infected 2-12 days postinoculation. The total number of worms recovered from the infected eggs was 163 or 22.5% of the 725 inoculated metacercariae. Eggs contained from 1 to 12 (average 5.6) worms per CAM. Worm length increased rapidly from an average of 0.5 mm at 2 days to about 3.0 mm at 6 days postinoculation. Ovigerous worms first were seen on day 8 PI, but fluke eggs did not develop embryos. Worm development in ovo lagged about 1 day behind that of in vivo worms. One worm maintained for 17 days on 2 successive CAMs reached 6 mm in length, contained about 100 eggs in its uterus, and laid an additional 100 eggs on the CAM surface.     

Further studies on Capillaria philippinensis: development of the parasite in the Mongolian gerbil

Capillaria philippinensis larvae from the digestive tract of a Northern Luzon freshwater fish (Hypselotris bipartita) experimentally exposed to embryonated eggs, were given by stomach tube to Mongolian gerbils (Meriones unguiculatus). The larvae developed into adults within 10 to 11 days and female worms produced larvae within 13 to 14 days. These larvae developed into second generation adults by days 22 to 24 and the second generation females produced eggs that were present in the feces of the animal on the average 26 days after infection. Most females were oviparous but a few larviparous females were always present. The gerbils died on an average of 46 days after infection, with the highest numbers of worms recovered between days 36 and 46. All stages of the parasite were generally found at necropsy. Gerbils developed patent infections after receiving 2 or 3 laeval from fish, and 852 to 5,353 worms were recovered at necropsy. These studies show that autoinfection in an integral part of the life cycle of C. philippinensis, both initially and in maintaining the infection. The natural transmission of the parasite was demonstrated when H. bipartita from a lagoon in the endemic area were fed to gerbils and 3 became infected. The parasite can also be maintained in the laboratory by transfer of worms by stomach tube from the small intestines of an infected gerbil to a clean gerbil.     

Inflammatory cell stimulation and wound healing in Sphaeridiotrema globulus experimentally infected mallard ducks (Anas platyrhynchos)

Thirty laboratory-reared mallard ducks (Anas platyrhynchos) were experimentally infected with Sphaeridiotrema globulus. Host cell-mediated immunity and wound healing in S. globulus infected ducks were evaluated by gross and histological examination. Establishment, location, and life span of S. globulus differed from previous reports of sphaeridiotremiasis in both naturally and experimentally infected waterfowl. No worms were recovered from the ceca, and worm migration occurred anterior to the ileo-cecal valve with greater dispersion (less worm crowding) at higher rates of infectivity. Parasite death and host lesion resolution were evident at days 8 to 10 postinfection (PI) in ducks fed a moderate dose (200 metacercariae, group A) with a 5% mean parasite recovery rate. Host death occurred at days 3 to 6 in ducks fed a high dose (550 metacercariae, group B) with a 16% mean parasite recovery rate. Mast cells increased significantly (P less than 0.005) in group A ducks from days 4 to 10 PI. Eosinophil proliferation was greater in group B than in group A on day 4 PI, but comparatively fewer eosinophils were identifiable in group B ducks on day 6 PI.     

Description and life-cycle of three new species of Dingularis n. g. (Digena: Plagiorchiida), parasites of Australian freshwater turtles

Dingularus n.g. is proposed to accommodate three new species, D. anfracticirrus, D. pearsoni and D. megapharynx, within the Plagiorchiida. Dingularus is closely related to the Plagiorchiidae, despite the unusual excretory systems of the three species. Adult worms were parasitic in the intestines of freshwater turtles, Chelodina expansa (D. pearsoni and D. megapharynx) and Emydura macquarii (D. anfracticirrus), and appeared to be host-specific. The three species had similar, three-host, aquatic life-cycles and each life-cycle was followed experimentally. The planorbid snail Glyptophysa gibbosa served as the first intermediate host for each species. Tadpoles of Limnodynastes peronii and snails G. gibbosa and Austropeplea lessoni served as second intermediate hosts. Dingularus spp. eggs were fully embryonated and infective when laid. They remained viable for 2-3 months in water but did not hatch until eaten by G. gibbosa. Miracidia were stimulated in the stomach of the snail host but did not hatch until eggs passed into the intestine. Hatching occurred in only the anterior fifth of the intestine. Hatched miracidia were not passed with the snail faeces. The pre-patent period in the snail differed in each species: D. anfracticirrus 42 days, D. pearsoni 23 days and D. megapharynx 32 days.     

Schistosoma ovuncatum n. sp. (Digenea: Schistosomatidae) from northwest Thailand and the historical biogeography of Southeast Asian Schistosoma Weinland, 1858

Schistosoma sinensium Bao, 1958 was first isolated from an unidentified snail in Sichuan Province, PR China. This species was apparently rediscovered in Chiang Mai Province, northwest Thailand (Baidikul et al., 1984); the definitive host was the rat Rattus rattus and the intermediate host was the snail Tricula bollingi. In this paper S. sinensium is rediscovered in Sichuan Province and compared with worms recovered from experimentally infected mice, which had been exposed to cercariae shed by T. bollingi from Chiang Mai. Evidence is presented suggesting that the schistosome collected by Baidikul was not S. sinensium and that a new species is involved. The new species, named Schistosoma ovuncatum (etymology: ovum (egg) + uncatus (hooked)), is described and compared with related taxa. All previous papers on the Thai schistosome have used worms recovered from field-collected rodents only; this is the first account in which the life-cycle has been completed in the laboratory, using cercariae shed by T. bollingi, and the resulting worms described. S. ovuncatum differs from S. sinensium in terms of size and shape of body and egg, number of testes, size of ovary, length of vitellarium, intermediate host and biogeographical distribution. The relationships of the two taxa and their position with respect to the Schistosoma indicum- and S. japonicum-groups are discussed. The implications of the findings for the evolution of human schistosomiasis in the region are also commented upon.     

Development and pathology of Echinostoma caproni in experimentally infected mice

In the present article, several parasitological features of mice, each experimentally infected with 75 metacercariae of Echinostoma caproni (Trematoda: Echinostomatidae), were studied during the first 12 wk postinfection. Moreover, the early pathological responses also were analyzed and compared with data previously published on other host species of E. caproni to gain further insight into the factors determining worm rejection or establishment of chronic infections. The results obtained show that the pattern of E. caproni infection in mice is consistent with a highly compatible host-parasite system. This combination is characterized by a high worm establishment, high egg output, and long survival of the worms. However, some differences with respect to other highly compatible hosts have been observed, particularly in relation to the survival of the adult worms. Histological studies suggest that the kinetics of goblet cells, mucosal neutrophils, and mononuclear inflammatory cells in the mesentery seem to be essential in determining the course of E. caproni infection in mice.     

Effects of a diet deficient in the B complex vitamins on infectivity, growth and distribution of Echinostoma caproni in ICR mice

The effects of a diet deficient in the B vitamins on infectivity, growth, and distribution of Echinostoma caproni in ICR mice were studied. The vitamin-deficient diet (experimental) was isocaloric to the control diet but lacked the B vitamins. Thirty-six female, 6- to 8-week-old ICR mice were each infected with 25 metacercarial cysts. From the day of infection to the day of necropsy, 18 mice were fed the experimental diet and the remaining mice received the control diet. Equal numbers of experimental and control mice were necropsied at 2, 3 and 4 weeks postinfection (p.i.). Mice on the experimental diet showed a significant loss in body weight between 2 and 4 weeks p.i. There was no significant difference in worm recovery at 2 to 4 weeks p.i. from mice on either diet. Worms from hosts on the experimental diet were more dispersed and located more posteriad in the small intestine than those from mice on the control diet. Worm dry weight was significantly less in hosts on the experimental diet at all weeks p.i. compared with that of hosts on the control diet. The body area of worms on the experimental diet was significantly less at 2 and 3 weeks p.i. than that of worms on the control diet. An isocaloric diet deficient in the B vitamins had a detrimental effect on the growth of E. caproni in ICR mice.     

Schistosoma japonicum infection in pregnant mice.

Ten 1-week and ten 2-weeks pregnant female NMRI mice were experimentally exposed to 70 Schistosoma japonicum cercariae. Ten littermice from each group were examined for worms by perfusion 4, 6 and 8 weeks post infection. Although the mothers (n = 15) were found infected with 15.5 +/- 13.4 worms at perfusion 6 and 7 weeks post infection, no worms were found in any of the examined littermice, as well as no detection of faecal or tissue eggs. Litter sizes did not differ from control groups and all littermice were healthy. The present study therefore suggests that congenital infection with S. japonicum does not occur in percutaneously infected mice and that infection of the mother during pregnancy does not seem to affect the offspring.