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Helminthic Infections of the Liver.

Human helminthic infestation is exceedingly common on a global scale, with as many as 1.5 to 2 billion people affected worldwide. Helminths (parasitic worms) that infect the liver and hepatobiliary system include nematodes (roundworms), cestodes (tapeworms), and trematodes (flatworms or flukes). The majority of morbidity and mortality from these infestations is caused by the host immune response to the larvae or adult worm. Helminthic disease manifestations vary from the extremes of asymptomatic carriage to cirrhosis and decompensated liver disease. Current basic science and clinical research focus on improvements in medical therapy, mass screening and chemoprophylaxis, and the development of preventative vaccine strategies.

Journal Article↗

Echinococcus multilocularis: characterization of a DNA probe.

A 0.6 kb DNA fragment has been isolated from a genomic sublibrary of the cestode Echinococcus multilocularis. This DNA-fragment showed a strong hybridization signal to 32PdCTP labeled total DNA prepared from E. multilocularis metacestode material. The fragment was subcloned into the Escherichia coli vector Bluescript BS+ resulting in the recombinant plasmid pAL1. The recombinant parasite DNA probe was labeled by biotinylation and hybridized to Southern blots of resolved EcoRI/PstI digested genomic DNA originating from E. multilocularis, E. granulosus and other helminth species (Taenia hydatigena, T. crassiceps, T. saginata, Mesocestoides corti, Hymenolepis diminuta, Moniezia expansa). The Southern blot hybridization experiments revealed that the DNA probe pAL1 was specific for E. multilocularis and E. granulosus. By comparison of the hybridization banding patterns a clear discrimination between E. multilocularis and E. granulosus was possible at the genome level. Furthermore, pAL1 was used to detect genetic variation within a set of different E. multilocularis isolates which had been experimentally maintained in mice by parasite tissue transplantation.

Animals↗

Serological survey of human cysticercosis in Irianese refugee camps in Papua New Guinea.

In 1984, over 10,000 refugees left the Indonesian province of Irian Jaya, and thus possibly imported Taenia solium taeniasis/cysticercosis into Papua New Guinea, which was believed to be free of T. solium until 1966. In a serological survey carried out in 1986, 50 refugees originating from areas endemic for T. solium and 171 patients from other areas with symptoms suggesting the possibility of cysticercosis were examined. As a sensitive prescreening technique an ELISA was used with a crude antigen extract obtained from T. solium metacestodes of pig origin. Of 221 persons investigated, 79 (36%) were positive in ELISA. For excluding frequently occurring cross-reactions in ELISA, Western-blotting (or EITB, enzyme-linked immunoelectrotransfer blot) was employed. In this test the demonstration of antibody activity to the 26 or the 8 kilodalton band has been proved to be species-specific for T. solium cysticercosis. One from 79 patients positive in ELISA was simultaneously positive (26 and 8 kDa) in Western blot, corresponding to the first case found in Papua New Guinea with a highly probable T. solium cysticercosis. This patient, originating from an endemic area in Irian Jaya, had immigrated into Papua New Guinea in 1980. The present work emphasizes the need for using highly specific immunodiagnostic techniques in seroepidemiology of larval cestode infections. T. solium taeniasis/cysticercosis remains a risk for Papua New Guinea, and refugees originating from endemic areas should be regarded as potential carriers of T. solium.

Animals↗

Recent advances in laboratory models for evaluation of helminth chemotherapy.

Extensive research on nematodes of veterinary importance has resulted in new tests based on incubation of parasitic nematodes in culture media, with a variety of detection systems including visual observation, inhibition of aggregation, reduced secretion of acetylcholinesterase and electronic motility measurements. There has been a proliferation of tests to detect benzimidazole resistant nematodes including egg embryonation, egg hatch, larval motility, differences in esterases, and tubulin binding by labelled benzimidazoles. A simple larval development test, which detects resistance to all types of anthelmintics, is likely to become the preferred test for evaluation of resistance in nematodes of grazing animals. By contrast the relative unimportance of trematodes and cestodes from a commercial point of view has resulted in few recent developments in models for evaluating chemotherapy. But the emergence of drug resistant strains of both schistosomes and liver fluke indicates the need for simple tests to detect resistance.

Animals↗

Partial purification and properties of ovine liver Echinococcus granulosus protoscolices phospholgucose isomerase.

Echinococcus granulosus protoscolex is the actual larval stage of the cestode causing echinococcosis both in man and animals. In the present report, certain properties of phosphoglucose isomerase from the ovine liver E. granulosus protoscolices have been studied and compared with those of the hydatid cyst fluid and the healthy ovine liver enzymes. The protoscolices enzyme prepared in a manner similar to the hydatid cyst fluid and the ovine liver enzymes exhibited the following properties: (1) pH optimum of 8.2 (2) KM value of 0.23 mM, (3) the enzyme was inhibited in the presence of high concentrations of alpha-D-glucose 6-phosphate, (4) no detectable inhibition of the enzyme was observed in the presence of phosphate ion up to 4.1 mM, (5) the protoscolices enzyme was less thermostable as compared to the hydatid cyst fluid and the ovine liver enzymes, (6) the protoscolices enzyme had a lower Ki value (0.7 mM) as compared to either the hydatid cyst fluid (1.1 mM) or the ovine liver enzymes (4.6 mM) when 6-phosphogluconic acid was used as a competitive inhibitor.

Animals↗

Entamoeba histolytica: inhibition in vitro by bithionol of respiratory activity and growth.

Endogenous and 2-propanol-supported respiration of intact trophozoites of of Entamoeba histolytica (stain HM-1:IMSS) were inhibited by bithionol, an effective chemotherapeutic agent for some trematode and cestode infections in humans. Dichlorophene and hexachlorophene also inhibited 2-propanol-supported respiration of the parasite. In contrast, ethanol formation by E. histolytica extract in the presence of N2 was scarcely inhibited by bithionol. The compound also inhibited in vitro growth of axenic (HM-1 strain) and polyxenic (strain HJ-1:KEIO) amoebae in culture. It took less than 24 hr to kill and disrupt virtually all amoebae of either strain with 0.28 mM bithionol. Omission of bovine serum from BI-S-33 medium resulted in considerably less disruption of HM-1 strain amoebae by the compound. However, organisms that looked undisrupted were strained with trypan blue. Moreover, the number of amoebae incubated for 10 min in the serum-free BI-S-33 medium containing 0.14 mM bithionol did not increase, even after incubation for 24 hr following replacement of the experimental culture fluid with fresh complete BI-S-33 medium free of the compound. These findings suggest that, although serum appears to diminish the antiamoebic action, some halogenated bisphenols (in particular bithionol) may be useful for treatment of amoebiasis.

1-Propanol↗

Taenia taeniaeformis: characterization of larval metabolic products and growth of host gastric cells in vitro.

Development of larvae of the cestode parasite Taenia taeniaeformis in the liver of rats induces gross hyperplasia of the gastric mucosa and excessive mucus production in the stomach without any direct contact with the stomach. Because the taeniid larvae are known to elaborate excretory-secretory (E-S) product in vivo and in vitro, the product was analyzed further, and its effects on cultured rat and dog stomach cells were investigated. In vitro E-S product contained less negatively charged glycosaminoglycan than either heparin or chondroitin sulfate, and proteins of various molecular weights. It stimulated the growth of both rat and dog stomach cells at concentrations of 3-9 micrograms protein/ml culture medium. At a concentration of 30 micrograms protein/ml culture medium, it stimulated hexosamine production in the cells up to 20 times, and multiple intracytoplasmic granules were found in both rat and dog cultured cells by light and electron microscopy. These results suggest that larval E-S product may be involved in the induction of gastric hyperplasia and hypermucus secretion.

Animals↗

Mesocestoides lineatus: trypsin induced development to adult mediated by Ca2+ and protein kinase C.

A mode of action of the inducible treatment with trypsin for the development of Mesocestoides lineatus tetrathyridium to adult was analyzed by administering various agents effective on Ca2+-dependent metabolic pathways in the cells: protein kinase C activators such as a synthetic diacylglycerol, 1-oleoyl-2-acetylglycerol, and a tumor promoting phorbol, 12-O-tetra-decanoyl-phorbol-13-acetate, enhanced the trypsin induced developmental processes. On the contrary, a calmodulin inhibitor, N-(6-aminohexyl)-5-chloro-1-naphthalene sulfonamide, cyclic adenosine 3',5'-monophosphate, and adenylate cyclase activators such as forskolin and cholera toxin, inhibited the triggering action of trypsin. Furthermore, a combined administration of Ca2+ ionophore (A23187) and the phorbol showed a similar effect with trypsin treatment, and sodium taurocholate acted as a potent enhancer like the activators of protein kinase C. These results strongly suggest that the initiation of development to adult in this cestode may be regulated synergistically by Ca2+ and protein kinase C, and that a bile acid may be involved in an activation mechanism of protein kinase C.

Calcimycin↗

Hymenolepis diminuta: intestinal goblet cell response to infection in male C57 mice.

Intestinal goblet cell numbers in two regions of the small intestine (20-30% and 60-70% distance form the pylorus) of male, 6- to 8-week-old C57 mice have been monitored following a 5-cysticercoid infection of the rat tapeworm, Hymenolepis diminuta. Test and sham-infected control mice were autopsied 0, 4, 8, 10, 14, and 28 days postprimary infection (p-1 degree-i) and 2, 4, 5, 7, and 14 days postsecondary infection (p-2 degree-i), administered 28 days p-1 degree-i. Results show a statistically significant increase in the number of mucus-containing goblet cells in both regions of the intestine during primary and secondary infections. Peak goblet cell numbers occurred on Day 8 p-1 degree-i and Day 5 p-2 degree-i in the 20-30% region and on Day 10 p-1 degree-i and Day 5 p-2 degree-i in the 60-70% region. In both regions, cell numbers declined to control levels by Day 14 p-1 degree-i, but remained significantly above control values 14 days p-2 degree-i. The increase in cell numbers correlated with an increase in goblet cell theca size and observable amounts of luminal mucus. The same infection regime in mice treated with cortisone elicited no goblet cell response. Male Wistar rats given a 10-cysticercoid infection and autopsied on Day 0, Day 10, and 15 months p-i showed a statistically significant increase in mucus-containing goblet cells only in the 60-70% region of intestine 10 days p-i; however, the worm burden was not eliminated. The functional significance of these results is discussed in relation to host immunity and murine cestode rejection.

Animals↗

Extraintestinal strobilar development of immature Echinococcus multilocularis in laboratory rodents following intratracheal inoculation of the protoscoleces.

Intratracheal inoculation of Echinococcus multilocularis protoscoleces was performed in prednisolone tertiary-butylacetate(PTBA)-treated and untreated golden hamsters and AKR/J mice. Repeated PTBA treatments prior to the day of inoculation prevented the influx of neutrophils against the cestode. More than 1 month after infection, living immature adult forms of E. multilocularis, showing some somatic prolongation, were recovered from the pulmonary alveolar space of PTBA-treated golden hamsters, in the absence of any cystic development.

Animals↗

Immunology and molecular biology of Echinococcus infections.

Echinococcus spp. are the etiological agents of hydatid disease in man and other intermediate hosts. Many questions regarding the factors which determine susceptibility/resistance to hydatid disease, and the factors which influence the viability and fertility of hydatid cysts, remain to be answered. Recent research into the effects of hydatid infection on the immune system of the host has provided some insights into the host-parasite relationships. Immunochemical and recombinant DNA techniques are being applied to improve diagnosis of hydatidosis in man and E. granulosus infection in dogs, and also in the development of vaccines against infection with taeniid cestode larvae. The successes which have been achieved in these areas are likely to provide valuable tools for the control of cystic hydatidosis in man. These recent studies in the areas of immunobiology, serological diagnosis and vaccination are reviewed.

Animals↗

Sparganum proliferum: an overview of its structure and ultrastructure.

A detailed study of the structure and ultrastructure of Sparganum proliferum was made possible for the first time thanks to the successful in vitro and in vivo maintenance of this rare parasite. Although S. proliferum exhibits many of the classical tegumental and parenchymal structures previously described for other larval cestodes, these are either arranged in a distinct fashion or, in some cases, may be completely different. Among the latter and of special interest are the single or multiple parenchymal cavities, surrounded by tegument, which in some instances appear to act as a primitive digestive tract.

Animals↗

Metabolism of lipid peroxidation products by the gastro-intestinal nematodes Necator americanus, Ancylostoma ceylanicum and Heligmosomoides polygyrus.

Somatic extracts of the three parasitic nematodes Necator americanus, Ancylostoma ceylanicum and Heligmosomoides polygyrus were able to detoxify a model hydroperoxide and a putative natural peroxide by glutathione-dependent peroxidase activity while cytotoxic carbonyls could be metabolized by NADPH-linked reduction activities. Unlike cestodes and digeneans, the nematodes in this study could not enzymatically conjugate carbonyls with glutathione. The results indicate that the three nematodes can protect themselves against possible host-immune initiated lipid peroxidation of their membranes at the level of the hydroperoxide and at the level of cytotoxic carbonyl, although other protective enzymatic mechanisms are also likely to exist (superoxide dismutase and catalase).

Ancylostoma↗

Intestinal contractions and migration behaviour in Hymenolepis diminuta.

The aim of this study was to determine if intestinal contractions were important in the migration behaviour of the rat tapeworm Hymenolepis diminuta. The objectives were to investigate the intestinal motility responses of the host to a meal which initiates worm migration, and the worms' responses to an artificial peristaltic contraction. A 1 g glucose meal elicited a significant orad migration by H. diminuta in the small intestine of the rat host when compared to water-fed controls (P less than 0.05). The glucose meal also significantly increased the transit rate, and thus, frequency of intestinal contractions in the small intestine of the rat, when compared to water-fed controls (P less than 0.05). Application of a circumintestinal ligature (6.3 g) (simulating an intestinal peristaltic contraction) resulted in significant worm migration when the ligature was applied in regions containing the worm's strobila as compared to controls where loose ligatures were tied in regions containing the strobila, or to controls where tight ligatures were tied ahead of the worm's strobila. These results suggest that H. diminuta migrates in an orad direction in response to the mechanical pressure produced by intestinal contractions induced by host feeding. It is concluded that contractions of the small intestine are an important cue in the migration behaviour of this cestode.

Animals↗

The effects of changes in the definitive host environment on the metabolism of Hymenolepis diminuta during growth and maturation.

Flexibility in the metabolism of Hymenolepis diminuta is associated with changing intrinsic requirements during maturation but is also influenced by extrinsic factors, that is, by the nature of the host environment. End-products of carbohydrate metabolism and enzyme activities in worm extracts were used as indicators of metabolic regulation in H. diminuta recovered at various times postinfection. The predominant end-product from 6-day-old worms is lactate, generated by cytosolic glycolysis. As the cestode matures in the host, lactate production by the whole worm decreases and greater amounts of the mitochondrial end-products, succinate and acetate, are detected. A stable, dichotomous carbon flow to lactate, succinate and acetate is observed from 12 days post-infection. A metabolic gradient along the length of individual strobila is also evident. It extends from glycolysis, in the anterior region, to mitochondrial dismutation in the posterior region. The transition from cytosolic to mitochondrial pathways during maturation and along the strobilus is delayed or suppressed in worms recovered from immunosensitized hosts. Four host environments were compared: unsensitized rats, rats immunosensitized with a primary infection of H. diminuta, rats immunosensitized with a primary infection of Nippostrongylus brasiliensis and mice concurrently infected with Heligmosomoides polygyrus. The specific activities of PK and PEPCK in whole worm extracts were similar in 10-, 21- and 35-day-old worms and did not differ in worms isolated from different host environments. However, the PEPCK/PK ratio is high in worms that utilize mitochondrial pathways and low in worms that produce predominantly lactate. LDH activity is high in lactate producers. It is concluded that the pattern of metabolism in H. diminuta is influenced by many effectors in the host environment.

Acetates↗

The phylogenetic position of the Aspidobothrea within the parasitic flatworms inferred from ribosomal RNA sequence data.

Sequence data from parts of the small subunit ribosomal RNA gene (18s rRNA) from an aspidobothrean and two digeneans have been added to a published data set which includes sequences from a number of parasitic and non-parasitic flatworms. Parsimony and distance matrix analyses of the data were carried out. Tests for particular clades were done using bootstrap resampling methods, and tests of particular topologies done using 4- and 5-taxon tests. The aspidobothrean Lobatostoma manteri emerges from near the base of the Neodermata, but whether as a sister group to the rest of the Neodermata or as a sister to the Digenea alone cannot be determined from the number of nucleotide sites available. Gyrocotyle is included with the cestodes in a very strongly supported clade in all analyses. There is evidence that the Monogenea is a paraphyletic group, although more data are needed to confirm this possibility.

Animals↗

Identification and partial characterization of the proteases from different developmental stages of Schistocephalus solidus (Cestoda: Pseudophyllidae).

The proteolytic activities of extracts prepared from procercoids and plerocercoids as well as adults of the pseudophyllidean cestode Schistocephalus solidus were examined using several proteins and synthetic peptides as substrates. Whole bodies of procercoids and the isolated syncytial tegument of plerocercoids and adults prepared by freeze-thaw were studied. Extracts of procercoids contained a chymotrypsin-like proteinase exhibiting a molecular weight of 23,500 determined by gel filtration chromatography. The proteinase showed collagenolytic activity and had a pH optimum at 8. Such a proteinase was absent in plerocercoids and adults. In these developmental stages leucine aminopeptidases were detected in the isolated syncytial tegument having molecular weights of 93,500 (plerocercoids) and 89,000 (adults), respectively. The aminopeptidases in both stages displayed optimal activity at pH 8.5. The chymotrypsin-like proteinase of the procercoid is possibly necessary for the penetration of the host's intestinal wall, whereas the aminopeptidases of the plerocercoid and the adult of S. solidus may aid in parasite nutrition by degrading oligopeptides at the tegumental surface.

Aminopeptidases↗

Evolutionary expansion of the Monogenea.

The evolutionary expansion of the monogeneans has taken place in parallel with the diversification of the fish-like vertebrates. In this article the main trends in monogenean evolution are traced from a hypothetical skin-parasitic ancestor on early vertebrates. Special consideration is given to the following topics: early divergence between skin feeders and blood feeders; diversification and specialization of the haptor for attachment to skin; transfer from host to host, viviparity and the success of the gyrodactylids; predation on skin parasites and camouflage; colonization of the buccal and branchial cavities; diversification and specialization of the haptor for attachment to the gills; phoresy in gill parasites; the development of endoparasitism and the origin of the cestodes; the success of dactylogyroidean gill parasites; the uniqueness of the polyopisthocotyleans; ovoviviparity and the colonization of the tetrapods. Host specificity has been the guiding force of coevolution between monogeneans and their vertebrate hosts, but the establishment of monogeneans on unrelated hosts sharing the same environment (host-switching) may have been underestimated. Host-switching has provided significant opportunities for evolutionary change of direction and is probably responsible for the establishment of monogeneans on cephalopod molluscs, on the hippopotamus and possibly on chelonians. There are indications that host-switching may be more common in monogeneans that spread by direct transfer of adults/juveniles from host to host. A limitation on the further expansion of monogeneans is the need for water for the dispersal of the infective larva (oncomiracidium).

Animals↗