Enteral and parenteral responses in mice after primary infection with Trichinella spiralis (Nematoda)

Enteral and enteral-parenteral infections were produced with T. spiralis in albino, Swiss Webster, outbred mice. Primary enteral infections abbreviated with thiabendazole stimulated inflammatory changes in Peyer's patches and the lamina propria of the small intestine of mice. These changes were accompanied by increased IgA in the intestinal luminal wash. Primary enteral-parenteral infections similarly stimulated the gut, and, in addition, the spleen. Splenic stimulation resulted in production of IgG1, and IgG2 antibodies specific for T. spiralis L3.     

Muscle fiber selectivity of Trichinella spiralis and Trichinella pseudospiralis (1983)

The biceps, semimembranosus, biceps femoris, and soleus muscles of female Rockland Wistar mice infected with either 1,000 Trichinella spiralis or 1,000 Trichinella pseudospiralis larvae were removed on days 12, 14, 16, and 18 post-infection (PI), sectioned and stained histochemically for their myosin ATPase activity. Light microscopic examination of the sections revealed that larvae of T. spiralis invade only the slow twitch muscle fibers, and those of T. pseudospiralis invade both the fast twitch and the slow twitch fibers. In sections obtained from mice infected with either parasite and killed on days 16 and 18 PI, identification of the majority of the infected fibers as fast twitch or slow twitch was not possible due to pathological modification of infected fibers.     

The role of malic enzyme in the carbohydrate metabolism of Trichinella spiralis spiralis and Trichinella spiralis pseudospiralis

The role of malic enzyme in the carbohydrate metabolism of Trichinella spiralis spiralis and Trichinella spiralis pseudospiralis. International Journal for Parasitology16: 435–440. The activities, intracellular localization and some regulatory properties of malic enzymes from homogenates of T.s. spiralis and T.s. pseudospiralis larvae have been studied. The malate saturation curves exhibit sigmoidicity. With increasing pH a ‘double sigmoidicity’ was observed in both NAD- and NADP-specific malic enzyme from T.s.spiralis and Trichinella malic enzymes resemble the Ascaris enzyme in their nondecarboxylation of oxaloacetate and in nucleotide and ammonium sulphate sensitivity, but the enzyme from T.s. pseudospiralis differs in its equal specificity for NAD and NADP. The cytoplasmic localizations and some properties of phosphoenolpyruvate carboxykinase in both Trichinella species are similar to the characteristics of the same enzyme in Ascaris.     

Trichinella spiralis in an agricultural ecosystem. II. Evidence for natural transmission of Trichinella spiralis spiralis from domestic swine to wildlife

Epidemiological investigations of an outbreak of trichinellosis were carried out in a domestic swine herd and it was established that the parasite also occurred in rats, and in skunks, opossums, and raccoons. Because considerable uncertainty exists regarding the role of sylvatic trichinellosis as a reservoir for the synanthropic cycle, studies were conducted to determine the genetic nature of the various isolates from this ecosystem. Pig infectivity trials, isoenzyme analyses, and repetitive DNA sequence analyses were performed. The results showed that all isolates from the farm environs were genetically similar and that they are related to Trichinella spiralis isolated from domestic pigs. The implication of these findings, in contrast to studies on isolates from wildlife elsewhere, is that this parasite is transmitted from domestic swine to sylvatic hosts and that any control or eradication efforts must take into account the potential for reinfection of hogs from wild animals.     

Characterization of endonuclease activity from excretory/secretory products of a parasitic nematode, Trichinella spiralis.

Double-stranded endonuclease activity was demonstrated for the first time in the excretory/secretory (ES) products of a parasitic nematode, Trichinella spiralis, which can reorganize host muscle cells. The endonuclease introduced double-stranded breaks to the native DNA. The ES double-stranded endonuclease(s) was sequence nonspecific, with a pH optimum below 6, and required divalent cations as a cofactor. Its activity was inhibited by the Zn2+ ion. It was detected mainly in the ES products of the infective-stage larvae of T. spiralis collected at 37 degrees C and was present in much smaller amounts in samples collected at 43 degrees C and in the products of T. pseudospiralis, a nonencapsulated species. The activity of endonuclease was blocked by antibodies against ES products. Zymographic analysis showed that the endonuclease activity was associated with at least three molecular forms, designated approximately 25, 30 and 58 kDa, respectively.     

The immune protection induced by a serine protease from the Trichinella spiralis adult against Trichinella spiralis infection in pigs

Trichinellosis is a major foodborne parasitosis caused by Trichinella spiralis. In the present study, a serine protease gene from an adult T. spiralis (Ts-Adsp) cDNA library was cloned, expressed in Escherichia coli and purified by Ni-affinity chromatography. Previous studies of our laboratory have found that mice vaccinated with recombinant Ts-Adsp protein (rTs-Adsp) exhibited partial protection against T. spiralis infection. In this study, the protective effect of rTs-Adsp against T. spiralis infection in pigs was further explored. The cell-mediated and humoral immune responses induced by rTs-Adsp were measured, including the dynamic trends of specific antibody levels (IgG, IgG1, IgG2a and IgM), as well as the levels of cytokines (IFN-γ, IL-2, IL-4, and IL-10) in the serum. Moreover, the changes in T lymphocytes, B lymphocytes, and neutrophils were measured to evaluate cellular immune responses in pigs vaccinated with rTs-Adsp. The results indicated that a Th1-Th2 mixed immune response with Th1 predominant was induced by rTs-Adsp after vaccination. Flow cytometric analysis showed that the proportions of CD4⁺ T cells, B cells, and neutrophils in the immunized groups were significantly increased. Furthermore, pigs vaccinated with rTs-Adsp exhibited a 50.9% reduction in the muscle larvae burden, compare with pigs from the PBS group five weeks after challenged. Our results suggested that rTs-Adsp elicited partial protection and it could be a potential target molecule for preventing and controlling Trichinella transmission from pigs to human.     

Characterization of cholinesterases from the parasitic nematode Trichinella spiralis

1. Trichinella cholinesterases occur in multiple molecular forms which differ in size, kinetics, activity with butyrylthiocholine, and effects of inhibitors.2. The 5.3 and 13S forms identified in Trichinella extracts are also found in C. elegans and other nematodes but the 7S form which occurs in other nematodes was absent from Trichinella detergent extracts. Differences in kinetic and inhibition properties among nematode species were also evident.3. The level of cholinesterases in excretory/secretory products is low.4. Trichinella cholinesterases did not elicit a detectable antibody response in mice.     

Hosts and habitats of Trichinella spiralis and Trichinella britovi in Europe

Trichinella spiralis and Trichinella britovi are the two most common species of Trichinella circulating in Europe. Based on data provided to the International Trichinella Reference Centre over the past 20 years (data referring to 540 isolates of T. spiralis and 776 isolates of T. britovi), we describe the host species and habitat characteristics for these two pathogens in Europe. A Geographical Information System was constructed using administrative boundaries, a Corine Land Cover (CLC) map, and an elevation map. In most countries, T. britovi is more widespread (62.5-100% of the isolates) than T. spiralis (0.0-37.5%), although in Finland, Germany, Poland and Spain, T. spiralis is more prevalent (56.3-84.2% of the isolates). Trichinella britovi is more widespread than T. spiralis in sylvatic carnivores (89% versus 11%), whereas T. spiralis is prevalent in both wild boars (62% versus 38%) and domestic swine (82% versus 18%), as well as in rodents (75% versus 25%). Trichinella spiralis and T. britovi circulate in the same environments: 41.1% and 46.0%, respectively, in agricultural areas, and 45.5% and 46.6% in forested and semi-natural areas. Although both pathogens can be transmitted by domestic and sylvatic cycles, their epidemiology is strongly influenced by the higher adaptability of T. spiralis to swine and of T. britovi to carnivores. These results are important because they include information on the countries at risk for these pathogens, the role played by specific species as reservoirs, the role of the pathogens in domestic and sylvatic cycles, and the role of the habitat in their circulation. The results can also be used to identify the most suitable animal species for the monitoring of these pathogens in Europe.     

Prevalence of Trichinella spiralis in black bears (Ursus americanus) from Newfoundland and Labrador, Canada.

Tongue and diaphragm samples from 158 black bears (Ursus americanus) from Newfoundland and Labrador were examined for Trichinella spiralis. No larvae were detected in samples from the island of Newfoundland but one animal from the Labrador samples was infected. The results of this and other studies suggest a lack of involvement of the black bear in a sylvatic cycle of T. spiralis in eastern Canada.     

Trichinella spiralis and breast carcinoma--a case report

Authors report about a patient with recurrent ductal invasive breast carcinoma and trichinosis. The patient underwent mastectomy of the left breast with evacuation of the axilla because of the cancer. Radiation therapy was received. An infestation with Trichinella spiralis was diagnosed two years after The patient was treated with mebendazole. A local recurrence of the tumor was found on the chest wall six years after the surgery. Tumor excision was performed. Histological analysis pointed at a ductal invasive carcinoma with numerous parasites of Trichinella spiralis present within both the muscle and the tumor tissue. The finding of parasites in the tumor tissue witnesses in favor of infestation, and the parasite morphology preserved in the tumor shows at the protective effects of the cysts, i.e. preventing parasite necrosis.     

Predilection Muscles and Physical Condition of Raccoon Dogs (Nyctereutes procyonoides) Experimentally Infected with Trichinella spiralis and Trichinella nativa

The predilection muscles of Trichinella spiralis and T. nativa were studied in 2 experimental groups of 6 raccoon dogs (Nyctereutes procyonoides), the third group serving as a control for clinical signs. The infection dose for both parasites was 1 larva/g body weight. After 12 weeks, the animals were euthanized and 13 sampling sites were analysed by the digestion method. Larvae were found in all sampled skeleton muscles of the infected animals, but not in the specimens from the heart or intestinal musculature. Both parasite species reproduced equally well in the raccoon dog. The median density of infection in positive tissues was 353 larvae per gram (lpg) with T. spiralis and 343 lpg with T. nativa. All the infected animals had the highest larvae numbers in the carpal flexors (M. flexor carpi ulnaris). Also tongue and eye muscles had high infection levels. There were no significant differences in the predilection sites between these 2 parasite species. Trichinellosis increased the relative amount of fat, but not the body weight in the captive raccoon dogs. Thus, Trichinella as a muscle parasite might have catabolic effect on these animals.     

A family history of deoxyribonuclease II: surprises from Trichinella spiralis and Burkholderia pseudomallei

Deoxyribonuclease IIalpha (DNase IIalpha) is an acidic endonuclease found in lysosomes and nuclei, and it is also secreted. Though its Caenorhabditis elegans homolog, NUC-1, is required for digesting DNA of apoptotic cell corpses and dietary DNA, it is not required for viability. However, DNase IIalpha is required in mice for correct development and viability, because undigested cell corpses lead to lesions throughout the body. Recently, we showed that, in contrast to previous reports, active DNase IIalpha consists of one contiguous polypeptide. To better analyze DNase II protein structure and determine residues important for activity, extensive database searches were conducted to find distantly related family members. We report 29 new partial or complete homologs from 21 species. Four homologs with differences at the purported active site histidine residue were detected in the parasitic nematodes Trichinella spiralis and Trichinella pseudospiralis. When these mutations were reconstructed in human DNase IIalpha, the expressed proteins were inactive. DNase II homologs were also identified in non-metazoan species. In particular, the slime-mold Dictyostelium, the protozoan Trichomonas vaginalis, and the bacterium Burkholderia pseudomallei all contain sequences with significant similarity and identity to previously cloned DNase II family members. We report an analysis of their sequences and implications for DNase II protein structure and evolution.     

Trichinella spiralis: a 76-kDa excretory/secretory larval antigen identified by a monoclonal antibody

Spleen cells from BALB/c mice were fused with myeloma cells following infection of the mice with Trichinella spiralis larvae and an ip booster injection with larval homogenate antigen. A monoclonal antibody (Mab), designated as TS 3G6 which did not react with sera or tissue extracts from noninfected mice, rats, and guinea pigs, was selected for further studies because of its high activity and specificity. When tested in ELISA TS 3G6 did not cross-react with Ascaris suum, A. lumbricoides, Toxocara canis, E. granulosus (larvae), Trichiuris suis, or T. ovis. Western blot analysis showed that Mab 3G6 recognized an antigen of 76 kDa located in the stichosome of the larvae as well as on the surface of the larval cuticle. Digestion of a larval extract with different enzymes suggests that the Mab TS 3G6 corresponding epitope is a polypeptide. The TS 3G6 antigen was detected in culture supernatants of Trichinella muscle larvae and in sera of experimentally infected animals using a sensitive ELISA assay. This secretory antigen also seemed to induce a specific immune response in the host since sera from infected animals could block the binding of Mab TS 3G6 to its target antigen when tested in a competitive ELISA.     

Extracellular vesicles from Trichinella spiralis: Proteomic analysis and protective immunity

Trichinella spiralis (T. spiralis) derived extracellular vesicles (EVs) have been proposed to play a key role in regulating the host immune responses. In this study, we provided the first investigation of EVs proteomics released by T. spiralis muscle larvae (ML). T. spiralis ML EVs (Ts-ML-EVs) were successfully isolated and characterized by transmission electron microscopy (TEM) and western blotting. Using liquid chromatograph mass spectrometer (LC-MS/MS) analysis, we identified 753 proteins in the Ts-ML-EVs proteome and annotated by gene ontology (GO). These proteins were enriched in different categories by GO, kyoto encyclopedia of genes and genomes (KEGG) and domain analysis. GO enrichment analysis indicated association of protein deglutathionylation, lysosomal lumen and serine-type endopeptidase inhibitor activity with proteins which may be helpful during parasite-host interaction. Moreover, KEGG enrichment analysis revealed involvement of Ts-ML-EVs proteins in other glycan degradation, complement and coagulation cascades, proteasome and various metabolism pathways. In addition, BALB/c mice were immunized by subcutaneous injection of purified Ts-ML-EVs. Ts-ML-EVs group demonstrated a 23.4% reduction in adult worms and a 43.7% reduction in ML after parasite challenge. Cellular and humoral immune responses induced by Ts-ML-EVs were detected, including the levels of specific antibodies (IgG, IgM, IgE, IgG1 and IgG2a) as well as cytokines (IL-12, IFN-γ, IL-4 and IL-10) in serum. The results showed that Ts-ML-EVs could induce a Th1/Th2 mixed immune response with Th2 predominant. This study revealed a potential role of Ts-ML-EVs in T. spiralis biology, particularly in the interaction with host. This work provided a critical step to against T. spiralis infection based on Ts-ML-EVs.     

Characterisation of novel protein families secreted by muscle stage larvae of Trichinella spiralis

Proteins secreted by Trichinella spiralis have a potential role in remodelling host skeletal muscle. However, whilst many parasite-secreted proteins have been identified, it has rarely been demonstrated that these are secreted into the nurse cell. Using an informatics-based analysis, we have searched the T. spiralis expressed sequence tag (EST) datasets for cDNAs encoding potential secreted proteins. Here we describe the characterisation of three of the top candidates isolated from our analysis, termed secreted from muscle stage larvae (SML)-1, -2 and -3. All three proteins were demonstrated to be secreted by muscle stage larvae, and immunohistochemical analysis established that SML-1 and -2 are secreted into developing nurse cells. We also show that SML-2 is processed from a precursor into smaller peptides by a metalloprotease contained within T. spiralis-secreted products. With the identification of these and other secreted proteins, we now have molecules to test in functional assays designed to dissect molecular features of the developing nurse cell.