Hymenolepis microstoma: direct life cycle in immunodeficient mice.

The mouse bile duct tapeworm Hymenolepis microstoma requires beetles as the obligatory intermediate host. However, when congenitally athymic NMRI-nu mice were infected with the mature tapeworm and allowed to eat their own faeces with tapeworm eggs, the oncospheres penetrated the intestinal tissue and developed to cysticercoids. After excysting, growth to adult worms occurs in the lumen of the small intestine and bile duct. Furthermore, the same happened when NMRI-nu mice, non-obese diabetic severe combined immunodeficiency (NOD/Shi-scid) mice and NOD/Shi-scid, IL-2 Rgamma(null) (NOG) mice were orally inoculated with shell-free eggs of this parasite. Differences between the cysticercoids of H. microstoma and H. nana developed in the mouse intestinal tissues were: (i) the time course for the development of fully matured cysticercoids of H. microstoma in mice was about 11 days but only 4 days for H. nana; and (ii) cysticercoids of H. microstoma developed in mice had a tail while those of H. nana had none.

The efficacy of a single intraperitoneal injection of oxolinic acid in the treatment of bacterial infections in goldsinny wrasse (Ctenolabrus rupestris) and corkwing wrasse (Symphodus melops) studied under field and laboratory conditions.

The efficacy of a single intraperitoneal injection of oxolinic acid to control an outbreak of atypical Aeromonas salmonicida infection in goldsinny wrasse (Ctenolabrus rupestris) and in the treatment of systemic vibriosis in corkwing wrasse (Symphodus melops) was examined. In addition a field study was performed to examine the effect of medication on the survival rate of goldsinny wrasse in Atlantic salmon cages. Four groups of wild caught goldsinny wrasse, each of 50 fish, were treated with an intraperitoneal injection of propylene glycol:saline (50:50) (control) or 50 mg/kg oxolinic acid at a concentration of 50 mg/mL. Three days after medication the fish in all groups were treated by an intraperitoneal injection of prednisolone acetate and an increase in seawater temperature from 9.0 to 11.5 degrees C. Cumulative mortalities were 18% in the two groups treated with oxolinic acid and 94 and 100% in the unmedicated control groups, giving a 'relative percentage survival' (RPS) value of 82%. A laboratory maintained population of originally wild caught corkwing wrasse experiencing high daily mortality was treated with oxolinic acid (50 mg/kg) or propylene glycol:saline (control). Cumulative mortalities were 84% (control) and 42% (oxolinic acid medicated group) giving an RPS value of 50%. In a field investigation using goldsinny wrasse approximately 30% were medicated with oxolinic acid (50 mg/kg) prior to stocking in cages with Atlantic salmon. In two of three cages the cumulative mortality was significantly lower (P = 0.025 and P < 0.001) in the medicated groups.

Anthelmintic screening of Zimbabwean plants traditionally used against schistosomiasis.

Extracts of 23 plant species used popularly against schistosomiasis in Zimbabwe were screened for their anthelmintic effect. Schistosomules of the trematode Schistosoma mansoni and cysticercoids of the cestode Hymenolepis diminuta were studied in vitro. The material consisted of 58 plant extracts, of which 37 killed the newly excysted cysticercoids within an hour, when incubated in a culture medium. Lethal concentrations varied from 0.8 to 103 mg/ml. All plant extracts showed activity against the tapeworms after 24 h. Ten of the best extracts were also tested against schistosomules. Five of these extracts showed activity. Lethal concentrations varied from 0.6 to 33.8 mg/ml of dry plant material. Extracts of stem and root from Abrus precatorius (Fabaceae), of root bark and leaves from Ozoroa insignis (Anacardiaceae) and of root bark from Zizyphus mucronata (Rhamnaceae) gave the best results against tapeworms. The best results against schistosomules were obtained with stem and root extracts from Abrus precatorius (Fabaceae) and stem bark from Elephantorrhiza goetzei (Mimosaceae). Although the activity of root and root bark extracts commonly used in traditional medicine was verified in this study, our results showed that also extracts from leaf and stem can be effective anthelmintics.

Age-dependent occurrence of the intestinal ciliate Balantidium coli in pigs at a Danish research farm.

A cross sectional study of the prevalence and intensity of Balantidium coli in pigs was carried out on a Danish research farm. The prevalence of B. coli infection increased from 57% in suckling piglets to 100% in most pig groups > or = 4 weeks old. The mean number of cysts per gram faeces (CPG) of pigs aged 12 weeks and younger were < or = 206, whereas pigs aged 28 weeks and > 52 weeks had significantly higher counts of > or = 865 CPG. Although some lactating sows had very high CPG's, no significant differences in CPG could be detected between the intensities of pregnant sows, lactating sows and empty and dry sows. No human cases of B. coli infection have been published in Denmark though it is zoonotic.

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.

Congenital transmission of Schistosoma japonicum in pigs.

Congenital transmission of Schistosoma japonicum in pigs was investigated by experimentally infecting sows at four weeks gestation (n = 3), 10 weeks gestation (n = 3), or a few weeks prior to insemination (n = 2). None of the piglets born to sows infected prior to insemination or in early pregnancy were found to be infected. However, all of the piglets (n = 26) born to sows infected at 10 weeks gestation were found to harbor schistosomes with S. japonicum eggs recovered from both their feces and livers. The findings show that congenital S. japonicum infection of pigs can occur if sows are infected during mid-to-late pregnancy and may have important implications not only for pigs but also for other mammalian hosts of schistosomes, including humans.

Immunology and biochemistry of Hymenolepis diminuta.

This review is an account of modern research into the immunology and biochemistry of the rat tapeworm, Hymenolepis diminuta. The first half of the review is devoted to the immunological responses of the host to the parasite. It describes the specific responses that occur when the host is exposed to a primary infection, and the changes that occur when further infections are superimposed on the primary one. The aquisition of immunity to the tapeworm and its persistence in the absence of the infection are also discussed, as well as the non-specific responses of the host to the parasite. The second half of the review is concerned with biochemistry, summarizing the early biochemical work that has been carried out on the tapeworm and describing the metabolic pathways now thought to be characteristic of the parasite. What little information that exists on intermediary metabolism in eggs and larvae is summarized here. Much of this section is concerned with the role of mitochondria in H. diminuta, especially the control of the critical branchpoint (PK/PEPCK), which partitions carbon into either the cytosol or the mitochondrion. The role of 5-hydroxytryptamine in controlling both worm behaviour and metabolism is discussed, followed by a brief look at some other effectors that may prove in the future to have great significance in regulating the parasite. Finally, there is a detailed consideration of strain variation within H. diminuta and of the impact on the tapeworm of components of the immune system, formerly described as the 'crowding effect'. The review concludes with a brief discussion of evolutionary aspects of the rat-tapeworm relationship and a comprehensive bibliography.

The contribution of hookworm and other parasitic infections to haemoglobin and iron status among children and adults in western Kenya.

A cross-sectional study of 729 children and adults in western Kenya investigated the impact of infection with hookworm, Ascaris lumbricoides, Trichuris trichiura, Schistosoma mansoni and malaria on iron status. In bivariate analyses, hookworm intensities as low as 300 eggs/g of faeces were negatively related to levels of haemoglobin (Hb) and serum ferritin (SF). Malaria parasitaemia was negatively related to Hb and positively related to SF, while S. mansoni intensities were negatively related to SF. Multivariate regression analysis was done to identify predictors of Hb and SF levels. In children, age (in years) was the only predictor for Hb (B = 1.7 g/L) and only malaria parasitaemia (negative, light, moderate, heavy) was retained in the model for log10 SF (B = 0.097 microgram/L). In adults, hookworm infection and malaria parasitaemia together with age, sex, pregnancy, SF levels < 12 micrograms/L and elevated body temperature were significant predictors of low Hb. The regression coefficient for hookworm egg count (for increments of 100 eggs/g) was -1.3 g/L. Significant interactions between sex and age and between sex and malaria parasitaemia were revealed. Age and malaria parasitaemia were significant predictors only among females, with a regression coefficient for malaria parasitaemia of -6.9 g/L. The regression coefficient for hookworm did not change when SF < 12 micrograms/L was taken out of the model, indicating that the effect of hookworm cannot be explained by low iron stores alone. Using SF as the dependent variable, hookworm and S. mansoni intensities together with age and sex were retained in the model. The regression coefficients for hookworm egg count (increments of 100 eggs/g) and S. mansoni egg count (increments of 10 eggs/g) were -0.011 microgram/L and -0.012 microgram/L, respectively. Iron deficiency was a problem in this population and hookworm infections contributed significantly to this situation.

Host-parasite relationships in infections with two Kenyan isolates of Schistosoma mansoni in NMRI mice.

To relate the marked differences observed in morbidity due to Schistosoma mansoni infection in the Kangundo and Kibwezi areas of Machakos District, Kenya with possible parasite-related differences in pathobiological characteristics, a quantitative comparison of the host-parasite relationship in infections in NMRI mice with S. mansoni isolates from the 2 areas was conducted. One hundred and 20 cercariae were used per mouse, and necropsy took place between 35 and 94 days following infection. Parameters assessed comprised worm establishment, tissue egg counts, fecal egg excretion, egg production/worm pair, and weights of liver and spleen. Central findings included a higher worm establishment and a lower egg production/worm pair in Kangundo infections than in Kibwezi infections, comparable total tissue schistosome egg counts, and a higher liver/intestinal tissue schistosome egg ratio in Kibwezi infected mice than in the Kangundo infected mice. In addition, a much higher fecal egg excretion, expressed as eggs/g feces and eggs/g feces/worm pair, was observed in mice infected with the Kangundo isolate of S. munsoni than in mice infected with the isolate from Kibwezi. Although possible limitations in the use of mouse models need be realized, the results provide some support to the hypothesis that parasite-related differences in pathobiological characteristics may at least contribute to the marked difference observed in S. mansoni-induced morbidity between the Kangundo and Kibwezi areas.

Interactions between intestinal tapeworms and their hosts: present knowledge and problems.

This review on interactions between intestinal tapeworms and their hosts covers our knowledge and problems regarding the life-span of adult tapeworms, chronic versus acute infections and their background, destrobilation and its causes, genetic influence on immunity to tapeworms, spontaneous cure and effects of host immunocompetence, antibodies and cell-mediated immune responses on intestinal tapeworms. Furthermore, evidence of protective antigens, acquired immunity, concomitant immunity and strategies adopted by tapeworms to circumvent host defenses are discussed.

In vitro maintenance of Schistosoma japonicum and surgical transfer from donor to naïve recipient pigs.

An objective of this study was to find a culture medium and a temperature range suitable for in vitro maintenance of adult Schistosoma japonicum during surgical transplantation experiments. Adult S. japonicum were cultivated in four different media (NCTC 135, NCTC 109, RPMI 1640 and 0.85% physiological saline) supplemented with 10% heat-inactivated normal pig serum (hiNPS) at either 4 degrees C, 22-25 degrees C (room temperature) or 37 degrees C. Based on survival and morphologic evaluation, NCTC 135 at room temperature was found to be the best medium/temperature combination for maintenance of worms. An additional objective was to develop a method for transplanting adult S. japonicum from experimentally infected donor pigs to naïve recipient pigs. Six Landrace/Yorkshire crossbred pigs were used as donors to supply worms for two recipient pigs. Worms for transplantation were obtained by perfusion of the mesenteric veins of the donor pigs and maintained for a maximum of 3 h in NCTC 135 + 10% hiNPS at room temperature. A total of 148 and 132 worms were surgically transferred by way of an infusion tube into caecal veins of the two recipients. Six weeks after transplantation, 14% and 36% of the transferred worms were recovered by perfusion and subsequent manual inspection of the mesenteric veins of the two recipient pigs, respectively. The successful results suggest that surgical transfer of S. japonicum worms from donor to naïve recipient pigs may be useful for future studies on population genetics, dynamics and regulation in the pig/S. japonicum model.

Anthelmintic usage of extracts of Embelia schimperi from Tanzania.

Embelia schimperi Vatke, belonging to the family Myrsinaceae, is used among the traditional Masai people of Tanzania and Kenya since it is believed to eliminate adult Taenia saginata, the beef tapeworm. In the present study, the anthelmintic effect of an extract of the dried fruits of Embelia schimperi was studied in experimental parasite/rodent models. In two experiments, rats with 10 adult tapeworms of Hymenolepis diminuta were treated with a diammonium salt of embelin (DE), isolated from the fruits. Significant lower numbers and total worm biomass of Hymenolepis diminuta were observed in rats treated with 100 mg DE/kg. Furthermore, clear indications of the occurrence of destrobilation was observed in faeces after treatment and in 16% of the worms found at autopsy. The killing effect shown in vivo was corroborated by in vitro studies, which showed that all adult Hymenolepis diminuta were killed when incubated in a culture medium containing as little as 0.08 mg DE/ml. No significant in vivo effect of DE was observed against Hymenolepis microstoma, the trematode Echinostoma caproni and the nematode Heligmosomoides polygyrus in mice, although the worms could be killed in vitro. These results indicate that the crushed seeds of Embelia schimperi taken orally by the Masai people indeed have an anthelmintic effect against human intestinal tapeworms.

Microscopical observations on immune precipitates formed in vitro on the surface of hymenolepid tapeworms.

Four-day-old worms of the tapeworms Hymenolepis microstoma, H. diminuta and H. nana and newly excysted H. microstoma were exposed in vitro at 37 degrees C to immune serum from mice infected for 4-12 weeks with H. microstoma. Worms were fixed for electron microscopy after intervals of 5 min to 96 h. Within 10-15 min an homogeneous precipitate occurred between the microtriches of 4-day-old H. microstoma and H. nana, while on some areas of H. microstoma the precipitate extended distal to the microthrix border and contained small vesicles (30 nm in diameter) and shed microtriches. In H. diminuta precipitates were not found until 2 h post-incubation. The thickness of the precipitate and the number of small vesicles and shed microtriches increased with time after incubation. Since a similar precipitate occurred on worms kept in complement-depleted immune serum, antibodies alone may induce immune damage. The precipitate on newly excysted H. microstoma lacked microthrix fragments. After 48 h an extensive precipitate was found protruding from the rostellar glands on some H. microstoma, and within the culture vessel. Antibodies may therefore be complexing with tapeworm secretory products.

Immunity to adult cestodes: basic knowledge and vaccination problems. A review.

Immunity in mammals to intestinal cestodes has been reviewed using the normal final host infected with the tapeworms Hymenolepis diminuta in rats and H. microstoma and H. nana in mice as a model. Primary infections up to a certain level continue to live as long the host, while most worms in infections with larger doses are destrobilated and expelled. It has been argued that concomitant immunity against a superimposed infection exists in rats and mice infected with H. diminuta and H. microstoma, respectively, and suggested that it also takes place in humans infected with Taenia spp. Immunity to secondary infections after expulsion of a primary infection occurs, but immunological memory is rather short-lived, although depression of worm growth occurs for at least two third of the rat's life. Serum antibodies have been shown to produce a direct precipitate on the surface of cestodes in vitro, but a direct effect of antibodies in vivo or the relationship with e.g. host effector cells, like mast cells and eosinophils, is unknown. It has been shown that peritoneal exudate cells from rats are able to kill H. diminuta in vitro. Very little is known about the mechanisms of tapeworms to counteract host immunological responses, but the tegumental glycoconjugates and discoidal secretory bodies are possible candidates. Passive transfer of immunity by mesenteric lymph node cells has only been successful using cells from H. nana egg-infected mice and has shown that only short-lived proliferating cells are responsible for transferring immunity. Vaccination procedures and problems are discussed with special reference to E. granulosus in dogs.

Inhibition of growth of a tapeworm Hymenolepis diminuta in its normal host (rat).

The biomass of 8-day-old worms of Hymenolepis diminuta in secondary infections, administered to rats 3-10 days after chemotherapeutically expelling a primary infection, was 70-90% less, and the worms were more posteriorly distributed, than in naive controls. The strong depressive effect on growth waned rapidly over 2-5 weeks, but even in rats not challenged until 17 months later, worm growth was weakly depressed by 30%. The extent to which growth was depressed in a secondary infection was independent of the number of worms in the challenge but increased with number of worms in the immunizing infection up to four to eight worms. Further increase up to 64 worms had little effect. This suggests, as it is known that the biomass of worms in a rat reaches a maximum with infections of between five and 10 worms, that the change in the intestine is proportional to biomass, not number, of worms. It is argued that partially suppressed immuno-inflammatory changes in the intestine, which will affect secondary worms so strongly, will also have depressed growth and fecundity effects on the primary worms, that a dynamic equilibrium is reached between the strength of the intestinal response and the biomass of the tapeworm, and that it is reaching this equilibrium, not a 'crowding effect', which limits H. diminuta to a level compatible with the survival of the rat.

Antagonistic effects of Schistosoma mansoni on superimposed Hymenolepis diminuta and H. microstoma infections in mice.

Patent, but not prepatent, Schistosoma mansoni infections in mice enhanced the expulsion of a superimposed infection with Hymenolepis diminuta. An antagonistic effect was also directed against a superimposed H. microstoma infection in mice harbouring patent S. mansoni infections.

Complement-mediated leucocyte adherence to newly excysted and 4-day-old Hymenolepis diminuta.

Four-day-old and newly excysted H. diminuta were exposed in vitro (37 degrees C; 5% CO2/air atmosphere) to serum and peritoneal cells (1.7-4.4 X 10(5] obtained from rats. Four-day-old worms incubated in serum alone were lysed in titres of less than 16. In assays containing peritoneal cells, leucocytes, predominantly eosinophils and macrophages, adhered to the posterior end of the parasite in serum titre 32, but not in serum titres 64 and 128. In this region of the worms phagocytosis of microtriches by macrophages, microthrix denudation and loss of tegument were noted. Serum-mediated lysis of newly excysted cysticercoids occurred at a serum titre of 64 and leucocyte adherence and phagocytosis of microtriches occurred in serum titres 128 and 256. Attachment of peritoneal cells to worms did not occur in assays containing heat-inactivated serum and it is suggested that regional leucocyte adherence and subsequent parasite damage is complement-mediated.

Concurrent infections of the trematode Echinostoma caproni and the tapeworms Hymenolepis diminuta and Hymenolepis microstoma in mice.

Superimposing the intestinal tapeworm Hymenolepis diminuta on an established infection with the trematode Echinostoma caproni or simultaneous infection of mice with H. diminuta and Hymenolepis microstoma caused destrobilation and expulsion of H. diminuta, whereas establishment and growth of H. microstoma under the same infection regimes were not affected. In contrast, simultaneous superimposition of H. diminuta and H. microstoma on an established E. caproni infection caused destrobilation and expulsion of both H. diminuta and H. microstoma.