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In the present paper, we assess the relationship between the expression of IFN-γ and the development of clinical signs in Echinostoma caproni-infected mice. For this purpose, we studied the course of the infection in three mouse strains: ICR (CD-1®) (a host of high compatibility with E. caproni), BALB/c (a prototypical Th2 strain), and BALB/c deficient for IFN-γ mice (IFN-γ(-/-)). Infection in ICR mice is characterized by the elevated expression of IFN-γ and iNOS in the intestine concomitantly with the lack of clinical signs. In contrast, the infection was more virulent in BALB/c and IFN-γ-deficient mice that developed a severe form of the disease together with the absence of IFN-γ expression. The disease was more severe in IFNγ(-/-) mice in which the disease was lethal during the few first weeks of the infection. The analysis of different parameters of the infection in each host strain showed that most of the features were similar in the three mouse strains, suggesting the IFN-γ plays a central role in that protection against severe disease. Thus, IFN-γ seems to play a dichotomous role in the infection facilitating the parasite establishment, but it may also benefit mice since it protects the mice from morbidity and mortality induced by the parasite.

The goal of this study was to characterize and quantify the various neutral and polar lipid classes in the BALB/c mouse that are associated with Echinostoma caproni infection. Ten infected mice and 10 uninfected control mice were used for this study (five infected and five uninfected were used for each of the neutral lipid and polar lipid studies). After 3 weeks postinfection, the mice were necropsied and various organs were removed and prepared for lipid class analysis. The organs used were liver, kidney, spleen, colon, cecum, anterior portion of the small intestine (SI), middle portion of the SI, and posterior portion of the SI. Lipids were determined by high-performance thin-layer chromatography (HPTLC) with Analtech 10 × 20 cm HPTLC-HLF silica gel plates. For neutral lipids, petroleum ether-diethyl ether-glacial acetic acid (80:20:1) mobile phase and 5% ethanolic phosphomolybdic acid detection reagent were used to determine the neutral lipids in each organ. Chloroform-methanol-deionized water (65:25:4) mobile phase and 10% cupric sulfate in 8% phosphoric acid detection reagent were used to determine the polar lipids in each organ. The analyzed polar lipids in all organs were phosphatidylcholine (PC), phosphatidylethanolamine (PE), and sphingomyelin (SM). Using HPTLC-densitometry for quantification, PC was found in the greatest amount and SM the smallest of all organs analyzed. The PE in the anterior portion of the SI was determined to be significantly greater (using the Student’s t test with P < 0.05), with about twice the amount of PE in mice infected with E. caproni relative to the uninfected mice. No significant differences in any of the neutral lipid classes were found between infected and uninfected samples.

The excretory/secretory proteome of Echinostoma caproni (Trematoda: Echinostomatidae) adults collected from experimentally infected mice was investigated using a proteomic approach. We performed a shot-gun liquid chromatography/tandem mass spectrometry for the separation and identification of tryptic peptides from the excretory/secretory products of E. caproni adult worms. Database search was performed using MASCOT search engine (Matrix-Science) and ProteinPilot software v2.0 (Applied Biosystems). A total of 39 parasite proteins were accurately identified. Strikingly, metabolic enzymes, and particularly glycolytic enzymes, constituted the largest protein family in the excretory/secretory proteome of E. caproni adult worms. Moreover, representative proteins involved in parasite structure, response against stress, chaperones, calcium-binding, and signal transduction were also identified. This work extends our knowledge of host-parasite relationships in the E. caproni-rodent model that is extensively used to analyze the factors determining the intestinal helminth rejection. Consequently, information on many proteins may be useful to better understand the molecular basis that determines the survival of this parasite in the definitive host.

This review examines the literature on imported (allochthonous) and local (autochthonous) cases of food-borne trematode (FBT) infections in the United States of America (USA) from 1890 to 2009. Most of the literature is concerned with imported cases of the opisthorchiids Clonorchis sinensis and Opisthorchis viverrini. These flukes cause serious pathology in the liver and biliary system of humans. Chronic cases may induce liver (hepatocarcinoma) and bile duct (cholangiocarcinoma) cancers in humans. Clonorchiasis and opithorchiasis are preventable diseases that can be avoided by eating properly cooked freshwater fish products. Several species of lung flukes in the genus Paragonimus are local or imported FBT in the USA. The endemic cycle occurs in the USA with various local snails and crustaceans serving as intermediate hosts. Paragonimids are acquired when humans eat raw or improperly cooked freshwater crustaceans containing metacercarial cysts. Infection can cause severe lung disease and the symptoms of paragonimiasis often mimic those of tuberculosis and other non-helminthic diseases. Paragonimiasis can be avoided by not eating raw or improperly cooked shellfish. The liver fluke Fasciola hepatica can be acquired by eating raw or uncooked vegetation. The cycle exists in the USA involving local snails and aquatic vegetation. Although some cases are local, most are imported by travelers or immigrants. Fascioliasis can cause serious liver and biliary diseases in humans and consumption of tainted vegetation should be avoided. Lesser known FBT have been reported in the USA including species of Alaria, echinostomids, heterophyids, troglotrematids, and a self-induced infection of Plagiorchis. Treatment of the FBT mentioned in this review consists of various regimens of praziquantel, except for F. hepatica where the drug of choice is triclabendazole.

This review updates an earlier one by Fried (Adv Parasitol 33:91-144, 1994) on metacercarial excystation of trematodes. Our coverage of the salient literature is mainly from 1995 to 2008 and includes significant studies on echinostomatid and non-echinostomatid trematodes. Most of the echinostomatid studies are on species of 37-collar spined Echinostoma, although one study is concerned with a species of Echinochasmus. The non-echinostomatid coverage includes work on species of Paragonimus, Haplorchis, Apatemon, Microphallus, Clonorchis, Riberoia, Maritema, Opisthorchis, and Zygocotyle. Most of the studies were concerned with methods used to excyst the metacercariae of the species being investigated. However, some studies were concerned with metacercarial excystation in order to use the excysted larvae for in vitro cultivation or morphometric studies. Other studies used the excysted larvae for physiological, biochemical, or behavioral work. Several studies were also concerned with elucidating both intrinsic and extrinsic factors which mediate chemical excystation of the metacercarial stage of digenetic trematodes.

This review is mainly concerned with the methods used to study collar spines in echinostomatid trematodes. The salient studies concerned with light microscopy on live and fixed larval and adult echinostomes are reviewed. Similar information is also provided for echinostomatid collar spines examined by scanning electron microscopy and transmission electron miscroscopy. The limited information on specific histochemical studies on these spines is also reviewed. Lastly, an examination of the few papers that have considered structure and function of collar spines is given.

This review updates our previous one (Reddy and Fried, Parasitol Research 100: 921-927, 2007) on Crohn's disease and helminths. The review considers the most recent literature on Trichuris suis therapy and Crohn's and the significant literature on the use of Necator americanus larvae to treat Crohn's and other autoimmune disorders. The pros and cons of helminth therapy as related to autoimmune disorders are discussed in the review. We also discuss the relationship of the bacterium Campylobacter jejuni and T. suis in Crohn's disease. The significant literature on helminths other than N. americanus and T. suis as related to autoimmune diseases is also reviewed.

Silica gel high performance thin layer chromatography-densitometry was used to determine the neutral lipid profile in the urine of BALB/c mice experimentally infected with adults of Echinostoma caproni. An approximate 30 worm burden per host induced certain changes in the neutral lipid profile of the mice at 2 to 10 weeks post-infection (PI) relative to the uninfected controls. Infection caused a significant increase (Student's t test, P < 0.05) in the methyl oleate fraction at 6 and 7 weeks PI relative to the uninfected controls. Neutral lipid profiles of urine may serve as an indicator of experimental infection of echinostomes in mice.

High-performance liquid chromatography (HPLC) was used to determine the effects of a patent Schistosoma mansoni infection on certain carboxylic acids in the digestive gland gonad complex (DGG) and hemolymph of Biomphalaria glabrata. An analysis of DGG samples was done using extraction with 50% Locke's solution, cleanup of the extract by anion exchange solid phase extraction (SPE), and ion exclusion HPLC with ultraviolet detection. Hemolymph was applied directly to the SPE column. Acetic, fumaric, malic, and pyruvic acids were detected, confirmed, and quantified at concentrations ranging from 12 to 280 ppm in the DGG and less than 124 to 8,000 microg/dl in the hemolymph. Infection with S. mansoni caused a significant reduction (Student's t test, P < 0.05) in the concentrations of acetic, fumaric, malic, and pyruvic acids in the DGG but not the hemolymph of B. glabrata compared to uninfected cohort snails. The significant reduction of certain carboxylic acids in the DGG of B. glabrata patently infected with S. mansoni suggests that these acids are utilized by the sporocysts and cercariae in the snail tissue, or that the infection stimulates reduced production or increased utilization by the snail tissue.

Echinostomes are intestinal trematodes with life cycles that are easy and inexpensive to maintain in the laboratory. For this reason, echinostomes have served for years as experimental models in different areas of parasitology. However, the usefulness of these trematodes in experimental parasitology has been under estimated. In this paper, we discuss the life cycles of echinostomes and the techniques used to maintain them in the laboratory. We further examine the characteristics of these trematodes that make them useful models for the analysis of larval parasite-host relationships. Echinostomes are useful for studies on the immunobiology of snails, host-finding processes of free-living larval stages, effects of larval trematode infections on anuran populations, and studies on analytes in the larval trematodes and their snail intermediate hosts.

Echinostoma caproni metacercarial cysts often need to be stored for several months to a year to meet laboratory requirements. This study examined the bacteria associated with encysted metacercariae stored at 4 degrees C. Bacterial assemblages from the kidney-pericardial region of uninfected snails and assemblages from the site of metacercarial infection in Biomphalaria glabrata were determined. Four species of Gram-negative rods were isolated and identified from the stored cysts to species or genus as Pseudomonas sp., Alcaligenes sp., Enterobacter cloacae, and Aeromonas hydrophila. Gram-positive rods were also isolated and included small slender rods (alpha hemolytic), small coccobacilli (gamma hemolytic), and small rods that exhibited gamma hemolysis. Accumulation of these bacteria on the surface of encysted metacercariae probably contributed to a decreased cyst viability of E. caproni.

The Echinostoma caproni-mice system is extensively used as an experimental model for the study of the factors involved in the establishment of chronic intestinal helminth infections. Although several parameters of the immunobiology of the host-parasite system have been studied in detail, the current knowledge of the cellular responses in these infections is still scarce. In the present paper, we analyze the kinetics of the circulating CD3(+) and CD19(+) cell populations and the different T-cell phenotype profiles in mice experimentally infected with E. caproni. Whereas the CD3(+) populations remained stable during the complete experiment, a marked increase in CD19(+) cells was observed from 4 weeks post-infection and beyond. Similarly, a marked increase in CD8(+) cell populations was observed in the 2 week post-infection. Our results show that E. caproni infection in mice alters the peripheral lymphoid cell populations, which may be important to determine the course of the infection. In this sense, CD8(+) cells can be essential in relation to their role as a source of IFN-γ.