Effect of water temperature on the development, release and survival of the triactinomyxon stage of Myxobolus cerebralis in its oligochaete host.

The development of the triactinomyxon stage of Myxobolus cerebralis and release of mature spores from Tubifex tubifex were shown to be temperature dependent. In the present work, the effect of temperature over a range of 5-30 degrees C on the development and release of the triactinomyxon stages of M. cerebralis was studied. Infected T. tubifex stopped releasing triactinomyxon spores 4 days after transfer from 15 degrees C to 25 degrees C or 30 degrees C. Transmission electron microscopic examinations of the tubificids held at 25 degrees C and 30 degrees C for 3 days showed that all developmental stages degenerated and transformed to electron-dense clusters between the gut epithelial cells of T. tubifex. In contrast, tubificid worms held at 5 degrees C and 10 degrees C examined at the same time were heavily infected with many early developmental stages of triactinomyxon. At 15 degrees C, the optimal temperature for development, maturing and mature stages of the parasite were evident. Infected T. tubifex transferred from 15 degrees C to 20 degrees C stopped producing triactinomyxon spores after 15 days. However, 15 days at 20 degrees C was not sufficient to destroy all developmental stages of the parasite. When the tubificid worms were returned to 15 degrees C, the one-cell stages and the binucleate-cell stages resumed normal growth. It was also demonstrated that T. tubifex cured of infection by holding at 30 degrees C for 3 weeks and shifted to 15 degrees C could be re-infected with M. cerebralis spores. The waterborne triactinomyxon spores of M. cerebralis did not appear to be as short-lived as previously reported. More than 60% of experimentally produced waterborne triactinomyxon spores survived and maintained their infectivity for rainbow trout for 15 days at water temperatures up to 15 degrees C. In natural aquatic systems, the triactinomyxon spores may survive and keep their infectivity for periods even longer than 15 days.     

Life history and population biology of adult Acanthocephalus lucii (Acanthocephala: Echinorhynchidae)

The life history and population biology of adult A. lucii in perch, Perca fluviatilis L., from the Forth and Clyde canal, Scotland, was investigated during May 1979-September 1981. There was an annual cycle in the size of the parasite population; prevalence and abundance (+/- SE) were highest during late spring and summer (70-90% and 14 +/- 4.3 to 16 +/- 5.6 worms/fish, respectively) but declined during late autumn and reached a minimum during winter (50-60% and 2.1 +/- 0.9 to 3.2 +/- 0.6 worms/fish). Parasite maturation was associated with higher water temperatures during spring and summer and most shelled acanthors were probably produced during summer and fall. There was only 1 generation of A. lucii per year, although generations tended to overlap and individuals within each generation did not develop synchronously. The sex ratio of adults was initially near unity but favoured females in the later stages of the infection. The distribution of A. lucii among perch was highly aggregated and stomach content analysis suggested that this was partly due to heterogeneity in perch feeding behaviour. The negative binomial and Poisson lognormal models fitted the data on worm distribution. Seasonal changes in the degree of parasite aggregation were detected, but no conclusive evidence of density-dependent controls on parasite population growth was obtained.     

An attempt to devise a hypothesis of the seasonal maturation of helminths in the definitive host--the fish

The seasonal maturation of four species of helminths, Acanthocephalus clavula, A. lucii, Camallanus lacustris and Bunodera luciopercae, from the perch Perca fluviatilis is briefly described. It is noted that A. clavula, A. lucii and C. lacustris are found as mature worms in the intestine of the fish host throughout the year, whereas in Bunodera luciopercae there is a limited season of maturation. As a result of experimental studies it is suggested that B. luciopercae uses host gonadotrophin to initiate and stimulate the early phase of gametogenesis. The gonadotrophin may also activate the parasites' own endocrine system, which then takes over control of subsequent spermatogenesis and oogenesis. This formation is used for a hypothesis for seasonal maturation of helminths in the fish definitive host. It also provides a potential explanation for seasonal maturation of helminths of fish in polar and tropical zones of the world.     

Partial purification and characterization of cysteine proteinases from various developmental stages of Paragonimus westermani

1. During development of Paragonimus westermani, larvae develop during migration within the host, and adult worms feed on pulmonary tissues, causing significant pathology in the mammalian host. In this report acidic extracts of various developmental stages (metacercariae and worms at one, two and three months of development) were examined for cysteine proteinase activity. 2. A soluble thiol-dependent proteinase activity with a native molecular weight of approximately 20,000 was isolated and partially purified. 3. The enzymes purified from the various developmental stages of the parasite had maximal activity at acidic pH and showed inhibitor susceptibilities similar to the vertebrate acidic cysteine proteinases. 4. Enzymatic activity was stable at pH 5.0 for at least two days when stored at 4 degrees C. 5. It is suggested that these enzymes may be involved in the nutrition of these parasites and/or during penetration and lysis of the tissues.     

Autoradiographic analysis of Schistosoma mansoni migration in the NZ rabbit

The migration of larval Schistosoma mansoni was tracked by means of autoradiographic analysis in naive rabbits percutaneously exposed to L-(75Se) selenomethionine-labeled cercariae on serial intervals of 1, 2, 4, 6, 8, 10, 15, 20, 25, 30, 40 and 50 days post-infection. Autoradiographic foci were detected from the 1st day in the skin, up to the 15th day in the liver. Adult and mature worms were recovered either paired or not 60 days after infection, by perfusion of hepatic and mesenteric veins. Morphometric analysis under optical microscopy, showed that worms were within regular dimention limits as compared to adult worms harboured by other host species. These observations extend previous informations on the S. mansoni-rabbit association and clearly demonstrate the post-liver phase of S. mansoni life-cycle in this host.     

Schistosoma mansoni: reversible destruction of testes by procarbazine

1. The anticancer drug procarbazine is profoundly damaging to the testes of Schistosoma mansoni when administered at 200 mg/kg or more to the mouse host. Somatic tissues appear entirely unaffected. 2. Within 2 days the meiotic process is disrupted, and primary and secondary spermatocytes and spermatids are destroyed and replaced by amorphous granular material. 3. The testes regenerate within about 15 days, apparently from surviving spermatogonial resting cells near the germinal epithelium of the testis. 4. Livers of mice treated once 7 weeks earlier have numerous egg granulomas and give rise to many miracidia, suggesting that full testis function is regained. 5. Male worms given the drug at 200 mg/kg at 19 days of age and fixed 7 weeks later have on average one testis less than control worms, indicating that about 15% of immature testes are unable to regenerate, whereas mature worms given the drug regenerate the normal number of testes. 6. The drug did not have significant antispermatogenic effects when male worms were incubated for a week with various concentrations in vitro, suggesting that a host metabolite is the active agent.     

Ontogenetic development of the brain of the platyhelminth Fasciola hepatica

During ontogenetic development in the definitive host, the cerebral ganglia of the parasitic flatworm Fasciola hepatica lose their cell rind integrity and develop specialized nerve processes. The organization and cytological features of the central nervous system were examined during three developmental stages in the parasitic life cycle of F. hepatica to determine when the changes occur. The cerebral ganglion cell bodies of migrating juvenile worms (5 days post-infection) are organized into a one-cell-thick rind that surrounds a central neuropile composed of small unmyelinated nerve processes (less than 3 microns in diameter). In young, sexually-immature adult worms (30 days post-infection), the cell bodies of the ganglia are no longer organized into a complete or tight cell rind around the ganglia. In addition, large diameter ('giant') unmyelinated nerve processes (greater than 12 microns) are found in the neuropile area. These giant nerve processes are also found in the transverse commissure and the longitudinal nerve cords. In mature adult worms (4-6 months post-infection), the rind of nerve cell bodies has completely disappeared and cell bodies are scattered around and within the neuropile. More than half of the volume of the mature adult neuropile is composed of giant nerve processes. The three developmental stages of the parasite that were used in this study differ significantly in their sizes, behaviours and microhabitat locations in the host. The results suggest that the organizational and morphological changes in the ganglia reflect selective adaptations to changes in the parasitic microenvironment.     

Recovery of adult stages and microfilaraemia after low dose inoculation of third stage larvae of Litomosoides carinii in Sigmodon hispidus

Because of the negative binomial distribution of filarial third stage larvae (L3) in their vectors, under natural conditions only a few are usually transferred per bite. After an inoculation of 5 L3 per animal into eight host animals at least one developed a long lasting patency based on one reproductive female only. After an inoculation of 15 L3, three of eight animals developed long lasting patency, harbouring between two and five fertile females. The rates of adult stages recovered were 0.43 and 0.30 respectively. The parasitaemias of the six patent animals in both experimental groups increased with the number of reproductive females present (r = 0.89, p = less than 0.005). All non-patent animals which were mf-negative in the pleural fluid and lung blood as well had a single sex or no worm load. In only one animal was there an apparently normal but non-reproductive pair of worms without any pathological alterations of the host animal. Encapsulated adult worms were found rarely, but independent of the final worm load or inoculation dose and always beside normal adults. In three of the 16 animals inoculated with 5 or 15 L3 patency passed after 10-12 weeks p.i., in two others it seemed to pass soon. After inoculation of 30, 40 or 60 L3 per animal patency passed early in about one half of 105 animals, when they were observed up to 24-36 weeks p.i. In conclusion all types of host defensive reactions are already visible after inoculation with such small doses.(ABSTRACT TRUNCATED AT 250 WORDS)     

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.     

Competition for minerals between Acanthocephalus lucii and its definitive host perch (Perca fluviatilis)

Concentrations of various essential and toxic elements (Ba, Ca, Co, Cu, Fe, Mg, Mn, Sr, Zn and Al, Ag, Cd, Cr, Ni, Pb, Tl) were analysed by inductively coupled plasma mass spectrometry in the acanthocephalan Acanthocephalus lucii and in different tissues of its host Perca fluviatilis. Nearly all the elements were found in significantly higher concentrations in the acanthocephalan than in the host tissues. Spearman correlation analysis revealed several strong inter-element associations within the organs of perch and within the parasites. Furthermore, statistical analysis revealed different competitive interactions. The concentrations of several essential elements (Ba, Ca, Fe, Mn, Sr and Zn) within the parasites decreased with an increasing number or weight of worms inside the intestine of the host. Additionally, the levels of some elements in the perch liver were negatively correlated with the weight of A. lucii in the intestine. Thus, it emerged that not only is there competition for essential elements between acanthocephalans inside the gut but there is also competition for these elements between the host and its parasites.     

The life history and biology of Diceratocephala boschmai (Platyhelminthes; Temnocephalida), an ectosymbiont on the redclaw crayfish Cherax quadricarinatus

Adults of the temnocephalan Diceratocephala boschmai, an ectocommensal on the crayfish Cherax quadricarinatus, deposited eggs on the host carapace. After 15 to 20 days at 28 °C, the eggs hatched into ciliated miniature adults with undeveloped reproductive organs, that remained attached to the host alongside the eggs. They grew and became gravid within 53 to 70 days if they were not dislodged. Most oviposited on the host on which they had hatched. Although larval temnocephalans have been described from in vitro work, this is the first temnocephalan life history to be described in vivo.Juvenile worms frequently changed their position on the carapace. Adult worms were sedentary just prior to, and during oviposition, and were frequently found at the base of the large chelae, the ventral surface of the antennae and antennules, and at the base of the walking legs. The ventral margins of the abdomen, the mouthparts, and the interorbital-rostral area were secondarily inhabited.It was calculated using an indirect regression that adult Diceratocephala boschmai survived on average 91 days when removed from the host but they did not produce eggs in vitro. On the host, adult worms lived an average of 48 days after the commencement of oviposition.Abbreviations C cilia - E eyespot - Es ejaculatory sac - Ex excretory canal - G gut - Gl glands - I intertentacular flange - M muscular peduncle - O ovary - P penis - Pg pharyngeal glands - Ph pharynx - R rhabdites - S seminal vesicle - T tentacles - Te testes - U uterus - Vd vas deferens - Vit vitelline glands     

Echinostoma revolutum: resistance to secondary and superimposed infections in mice

A complete or almost complete resistance (94-100%) to a superimposed Echinostoma revolutum infection existed in mice harboring 20-, 30-, and 40-day-old infections in the range of 2-4 to 30-35 worms, but no resistance was found at challenge Day 10. A similar high level of resistance (85-100%) also existed in mice for at least 6 weeks after natural expulsion of a primary 6 metacercarial infection and for at least 5 weeks after anthelmintic termination of a 30-day-old 20 metacercarial infection. Thymus-deficient nude mice failed to develop resistance to a superimposed infection, and the resistance in normal mice was inhibited by corticosteroid treatment. These findings are all in favor of a host immune response being responsible for the resistance against both a secondary and a superimposed infection. Nearly all the worms of a superimposed infection were, in resistant mice, expelled prior to 24 hr following infection (rapid expulsion), and the few worms circumventing this early expulsion persisted for at least 8 days. Newly excysted juvenile worms implanted intraduodenally into resistant mice were rejected to the same degree as juvenile worms from an oral metacercarial infection indicating that the newly excysted juvenile worms are the target of the host immune response. However, 7-day-old worms implanted intraduodenally into resistant mice survived indicating that adaptation to the host immune response had occurred. In conclusion, this host-parasite model is an example of concomitant immunity because the immunological mechanism responsible for the expulsion of the superimposed infection had no effect on the number of primary worms present.     

Cultivation of Clinostomum marginatum (Digenea: Clinostomatidae) metacercariae in vitro, in chick embryo and in mouse coelom

Metacercariae of Clinostomum marginatum obtained from naturally infected Perca flavescens were cultured by 5 methods. In vitro cultivation at 37 C in twice daily changes of Tyrode's, Na-poor Tyrode's, and Eagle's media failed to produce ovigerous adults after 4-5 days. Metacercariae placed on the chorioallantois of week-old chick embryos at 37 C migrated to the albumen. Only 8 of 123 worms recovered were ovigerous after 4-7 days in ovo. Best success occurred with metacercariae injected in groups of 4 or 6 into the abdominal cavities of male A/J mice. Despite liver lesions and strong inflammatory responses in the host, all 174 worms recovered were ovigerous after 4-8 days. None of 41 mice died prematurely from the procedure or the parasites.     

Growth and development of Metorchis orientalis in chicks and its adult morphology]

In order to observe the infectivity, growth and development and adult morphology of Metorchis orientalis, a total of 40 chicks were experimentally infected with 100 metacercariae respectively, collected from Pseudorasbora parva. The worms of various developmental stages were recovered from chicks at 1.5, 3, 5, 7, 9, 11, 14 and 21 days after infection, and they were prepared for morphological observations and measurements. All of the worms were found in the gallbladders of chicks, and their recovery rate was 32% in average. The growth of the body was rapid from 9 to 11 days after infection. The genital primordia appeared in 1.5 and 3-day old worms, and ovary and testes were first observed in 5-day old worms. Thereafter, genital organs gradually matured and completed up to 11 days after infection. The adult worm was leaf-like, and possessed a convoluted tubular seminal vesicle, an ovoid ovary, a sac-like seminal receptacle, 2 lobed-testes and follicular vitellaria. Eggs were 31.9 x 15.3 microns in average size, ellipsoid to elliptical in shape and possessed abopercular thickenings. From the above results, it is concluded that M. orientalis grows in sigmoid pattern in chicks, and their genital organs fully mature between days 9 and 11. It is also confirmed that a chick is a new definitive host of M. orientalis.     

Further studies on the effect of cyclosporin A on the course of Paragonimus infection in rats

Paragonimus ohirai-infected rats were treated with cyclosporin A (CyA) at different times during the course of infection. CyA (5 x 80 mg/kg) affected the worm recovery, growth and maturation rates of P. ohirai with respect to control values. This tendency was most remarkable in animals treated 15 days and more after infection with CyA (groups B, +15 to +19 days; C, +25 to +29; D, +35 to +39 and E, +45 to +49). In group A (0 to +4), however, the drug did not affect markedly the growth and maturation of worms, although it significantly lowered worm recovery rates. CyA administration also affected normal migration of P. ohirai in the highly susceptible host (rat), when the drug was administered during the peritoneal and/or liver phase of infection. Thus, in this P. ohirai/rat model, CyA significantly reduced worm recovery rates, and affected the growth, maturation and migration of the worms depending on the time of administration.     

Recovery rate, growth and development of Heterophyopsis continua in experimental chicks

The growth and developmental pattern of H. continua was observed after experimental infection of their metacercariae to chicks. The recovery rate of worms from the chicks at 1 to 28 days post-infection (PI) was 12.8% in average. The rate remained fairly high for early 4 days of infection but decreased thereafter rapidly till 28 days PI. Most of the flukes, 91.9%, were recovered from the ileum of the chicks. In metacercariae, genital organs such as the ovary, testes, seminal vesicle, seminal receptacle and genital sucker were recognizable. At one day PI Mehlis' gland appeared, and at 2 days follicular vitellaria were observed. At 3 days PI, eggs were formed in the uterine tubule and increased in number as the worm grew old. The worms reached 2,990 microns in length and 525 microns in width at 28 days PI. Genital organs developed rapidly in early stages of infection but slowly thereafter to 28 days PI, whereas non-genital organs developed steadily through the infection period. It was proved by this experiment that chicks should be a moderately suitable final host of H. continua.     

Residues effects of isoproturon in mature earthworm (Aporrectodea caliginosa) under laboratory conditions

This study was conducted to investigate the residues of isoproturon and its metabolites, 1-(4-isopropylphenyl)-3-methylurea, 1-(4-isopropylphenyl) urea, and 4-isopropylanilin in soil and mature earthworms under laboratory conditions. Mature earthworms (Aporrectodea caliginosa) were exposed for various durations (7, 15, 30, and 60 days) to soils contaminated with isoproturon concentrations (2, 4, 6, 8, and 10 mg.kg(-1) soil). The decrease in isoproturon concentration in the soil depended on initial concentration it was slower at higher concentrations. The isoproturon and its metabolites accumulated in earthworms it increased during the first 15 days and decreased thereafter. Acute toxicity of isoproturon was determined together with total soluble protein content and glycogen of worms. These parameters were related to isoproturon concentration in soil and earthworms. No lethal effect of isoproturon was observed even at the concentration 1000 mg.kg(-1) soil after 60 days of exposure. A reduction of total soluble protein was observed in all treated worms (maximum 59.54%). This study is suggesting the use of the total soluble protein content and glycogen of earthworms as biomarker of exposure to isoproturon.     

Proximate factors affecting the larval life history of Acanthocephalus lucii (Acanthocephala)

The growth and eventual size of larval helminths in their intermediate hosts presumably has a variety of fitness consequences. Therefore, elucidating the proximate factors affecting parasite development within intermediate hosts should provide insight into the evolution of parasite life histories. An experimental infection that resulted in heavy intensities of an acanthocephalan (Acanthocephalus lucii) in its isopod intermediate host (Asellus aquaticus) permitted the examination of parasite developmental responses to variable levels of resource availability and intraspecific competition. Isopods were infected by exposure to egg-containing fish feces, and larval infrapopulations were monitored throughout the course of A. lucii development. The relative rate of parasite growth slowed over time, and indications of resource constraints on developing parasites, e.g., crowding effects, were only observed in late infections. Consequently, the factors likely representative of resource availability to larval parasites (host size and molting rate) primarily affected parasite size in late infections. Moreover, at this stage of infection, competitive interactions, gauged by variation in worm size, seemed to be alleviated by greater resources, i.e., larger hosts that molted more frequently. The relatively rapid, unconstrained growth of young parasites may be worse for host viability than the slower, resource-limited growth of larger parasites.     

Influence of lactic acid bacteria on longevity of Caenorhabditis elegans and host defense against salmonella enterica serovar enteritidis

This study aimed to develop a convenient model to investigate the senescence of host defenses and the influence of food and nutrition. A small soil nematode, Caenorhabditis elegans, was grown for 3 days from hatching on a lawn of Escherichia coli OP50 as the normal food source, and subsequently some of the nematodes were fed lactic acid bacteria (LAB). The life spans of worms fed LAB were significantly longer than the life spans of those fed OP50. To investigate the effect of age on host defenses, 3- to 7-day-old worms fed OP50 were transferred onto a lawn of Salmonella enterica serovar Enteritidis for infection. The nematodes died over the course of several days, and the accumulation of salmonella in the intestinal lumen suggested that the worms were infected. The 7-day-old worms showed a higher death rate during the 5 days after infection than nematodes infected at the age of 3 days; no clear difference was observed when the worms were exposed to OP50. We then investigated whether the LAB could exert probiotic effects on the worms' host defenses and improve life span. Seven-day-old nematodes fed LAB from the age of 3 days were more resistant to salmonella than worms fed OP50 until they were infected with salmonella. This study clearly showed that LAB can enhance the host defense of C. elegans and prolong life span. The nematode appears to be an appropriate model for screening useful probiotic strains or dietetic antiaging substances.     

Influence of Lactic Acid Bacteria on Longevity of Caenorhabditis elegans and Host Defense against Salmonella enterica Serovar Enteritidis

This study aimed to develop a convenient model to investigate the senescence of host defenses and the influence of food and nutrition. A small soil nematode, Caenorhabditis elegans, was grown for 3 days from hatching on a lawn of Escherichia coli OP50 as the normal food source, and subsequently some of the nematodes were fed lactic acid bacteria (LAB). The life spans of worms fed LAB were significantly longer than the life spans of those fed OP50. To investigate the effect of age on host defenses, 3- to 7-day-old worms fed OP50 were transferred onto a lawn of Salmonella enterica serovar Enteritidis for infection. The nematodes died over the course of several days, and the accumulation of salmonella in the intestinal lumen suggested that the worms were infected. The 7-day-old worms showed a higher death rate during the 5 days after infection than nematodes infected at the age of 3 days; no clear difference was observed when the worms were exposed to OP50. We then investigated whether the LAB could exert probiotic effects on the worms' host defenses and improve life span. Seven-day-old nematodes fed LAB from the age of 3 days were more resistant to salmonella than worms fed OP50 until they were infected with salmonella. This study clearly showed that LAB can enhance the host defense of C. elegans and prolong life span. The nematode appears to be an appropriate model for screening useful probiotic strains or dietetic antiaging substances.