Amelioration of Myogenin, Bcl-2 expression and DNA damages in myocytes of Trichinella spiralis-infected mice after immunization with gamma radiation-attenuated larvae

iTRAQ-based proteomics of excretory-secretory products of Trichinella spiralis and Trichinella pseudospiralis at the muscle larva stage

The data of present study on the larval load of Trichinella-positive carcasses complement those of similar studies in pine marten and extend to wolf and fox species the identification of muscle districts predictive of the overall larval load of a carcass. In these species, the digestion of diaphragm and lower part of forelimb showed the best diagnostic capacity (the highest average value of LPG), respectively in the wolf and red fox. While the back district in the wolf and the rump in the red fox showed the highest predictive value for estimating the overall larval load of carcasses (the best r2 lpg values). In spite of the few number of examined animals and the unavoidable biases related to the variability of age, sex, carcass conditions and the same technique of muscle separation, the estimation of larval biomass of T. britovi infected carcasses, obtained by determining predictive LPGs and using the estimation of the overall muscle mass of an animal, is the basis for further studies on Trichinella contamination at the level of host populations. These studies should complement data from this study with those from the Trichinella host populations censuses. Knowledge of larval biomass at population level and its evolution over time may allow the adoption or strengthening of control measures in the territory based on the targeted recovery of host species' carcasses from the environment.

Experimental observation of the development of Trichinella spiralis muscle larvae in mice

Trichinella - a hairy and meaty tale.

Trichinella spiralis, Ascaris lumbricoides, and Taenia crassiceps are all parasitic helminths that can infect humans. After invading host, the parasites can induce immune responses such as activation of immune cells and induction of cytokines in the host. The immune responses induced by parasitic helminths have similarities with anti-tumor immune responses. Recent studies have demonstrated that infection of Trichinella spiralis, Ascaris lumbricoides, or Taenia crassiceps can inhibit the development of several kinds of tumors to some extent. In this paper, the progress of anti-tumor effects induced by the above-mentioned helminths on liver cancer, lung cancer, breast cancer, malignant melanoma, myeloma, and other cancers is reviewed, which may provide a valuable reference for treatment of tumors by the immune response to helminthic infection. Key words: Helminths; Infection; Anti-tumor activity

Introduction: Trichinellosis is a parasitic infection caused by the Trichinella genus of roundworms. The main route for contraction is eating raw or undercooked meat, especially pork. In the last report of the European Centre for Disease Prevention and Control Bulgaria and Romania account for 88% of the confirmed outbreaks in Europe with an average annual incidence of 0.8 per 100 000 for 2014. Materials and methods: A small outbreak of trichinellosis in 3 family members is presented. The re­search methods include an analysis of clinical history, physical examination, biochemical and sero­logical tests and enzyme trichinelloscopy of the meat. After the etiological treatment the development of the symptoms was monitored for the course of the hospitalisation and one month after that. Results: The initial signs of diarrhoea, nausea, abdominal pains, and high fever were treated on an ambulatory basis as originating from a flu infection. Upon the specialized physical examination mus­cle pains, periorbital oedema and macular rashes were observed which required immediate hospi­talisation. The family presented an anamnesis of consuming sausage, made of pork meat from a lo­cal farm. The laboratory tests showed extremely high numbers of eosinophils. The initial serological tests were negative, which is not unusual at this stage of trichinellosis. On the grounds of the national­ly adopted algorithm the combination of the clinical features, hypereosinophilia and a positive result for the consumed meat were sufficient for implementing a treatment course with Albendazole. The symptoms diminished during the hospitalisation period and no complications were observed upon the control examination. Conclusion: The diagnostic process of trichinellosis can be a challenging task as the mild forms are often mistaken for a flu infection. The difficulty in the diagnosis arises because the initial serologi­cal findings usually remain negative in the first weeks, but this moment is crucial for the etiological treatment to avoid the life-threatening complications as myocarditis or encephalopathy.

This review summarizes known data on the species composition of nematodes in 24 species of rodents inhabiting the territory of Armenia (South of Lesser Caucasus). The studied rodents include members of some cosmopolitan species (Mus musculus, Rattus norvegicus, Rattus rattus), as well as endemic and narrow-areal species (Meriones vinogradovi, Spermophilus xanthoprymnus, Microtus schidlovskii, Mesocricetus brandti, Sciurus anomalus). The taxonomical identity of the nematode species reported by previous authors is discussed with regard to present knowledge about helminth fauna of rodents. Our review concludes that 34 nematode species are present in rodents in Armenia, five of which are identified for the first time in rodents from the territory of Armenia, and three of which (Trichinella spiralis, Trichostrongylus colubriformis, Trichostrongylus skrjabini) are able to parasitize humans.

Pork meat and processed pork meat products may carry some hazards that threaten the consumer’s health. These hazards are related with animal health or with the way in which the raw materials are manipulated during slaughtering and manufacturing until the consumption. Substances having anabolic effects, the residues of veterinary drugs and chemical contaminants, some of them with origin in animal production but others produced during the transformation processes should be considered among chemical hazards. Concerning biological hazards, we can mention some parasites like Toxoplasma gondii, Trichinella spiralis, Taenia solium and also some bacteria like Salmonella spp., Campylobacter spp., Yersinia enterocolitica and Listeria monocytogenes. The physicochemical characteristics of fresh meat further facilitate the growth of various microorganisms, other than those mentioned above, which turns it into a highly perishable product. Several transformation processes of the raw material, provide different food products to consumers and constitute ways to increase its shelf-life. Drying, fermentation and cure are among these processing methods. In Portugal there is a wide variety of processed meat products, such as Chourico, Paio, Presunto, Salsichao and Catalao many of them manufactured according to traditional techniques of their regions of origin. Some of these products have been studied at the Universidade de Evora. Some results, which show the favourable effect of a range of transformation processes in controlling some of the mentioned biological hazards, will be presented.

Data set containing test results of screening blood and serum samples from five populations of Brown Bears in Alaska. Samples were tested to detect current or previous exposure to bacteria (Brucella spp., Francisella tularensis, and Leptospira spp.), viruses (canine adenovirus, canine distemper virus, canine parvovirus, and influenza A virus), and parasites (Toxoplasma gondii, and Trichinella spp.).

A parasitologic study is presented of 103 adult house rats from various sites in San Jose, Costa Rica. Seven specimens were Rattus rattus rattus, and the rest were Rattus norvegicus. The following protozoa were found in trophic form when feces were examined for eggs and larvae of helminths: Trichomonas, Giardia, Hexamita, Chilomastix, and Entamoeba. Trypanosoma lewisi was found in the blood of 7 out of 17 rats examined (8,8%) The following nematodes and cestodes were found with the incidence indicated: Strongyloides ratti (41,7 %); Aspiculuris tetraptera (1,9 %); Nipostron­gylus brasiliensis (47,6%); Gongylonema neoplasticum (0,9 %); Protospirura muris (8,7%); Trichosomoides crassicauda (23,3%); Taenia taeniformis larvae (33%); Hymenolepis diminuta (43,6%); and H. nana (0,9 %). No larvae of Trichinella spiralis were found in diaphragms examined. The acanthocephalian Moniliformis moniliformis was found in 18,4 per cent of the rats. Arthropods found as ectoparasites were, in arder of incidence, Echino­laelaps echidninus (14,5%), Xenopsilla cheopis (7,7 %) and Ctenopsyllus se­gnis (1,9%). These percent data should be regarded as minimum figures, as only 61 rats were brought alive to the laboratory.

A 35-year-old patient presented with a 3-month history of gradually progressive proximal muscle weakness of all four limbs. He had no constitutional features and no evidence of associated connective tissue disorder. Initial evaluation revealed raised muscle enzymes along with MRI and EMG evidence of idiopathic inflammatory myositis (IIM). Subsequently, muscle biopsy showed presence of trichinella larvae confirming a diagnosis of trichinella inflammatory myopathy (TIM). He was managed with anthelmintics and oral steroids and attained a good clinical recovery. Clin Infect Immun. 2016;1(1):8-10 doi: http://dx.doi.org/10.14740/cii4w

To observe the protective effects of recombinant trichinella spiralis-53 000 protein (rTsP53 protein) combining with imipenem (IMP) on septic mice and their underlying mechanisms. . Male BALB/c mice were divided into five groups randomly. Cecal ligature and puncture (CLP) operation was used for building polymicrobial septic model (CLP group).Mice in sham group were only subjected to laparoromy and abdominal closure without cecum ligation. At 6 hours post CLP, mice in CLP+IMP,CLP+rTsP53,and CLP+IMP+rTsP53 groups were injected intraperitoneally with IMP (20 mg/kg) + 0.1 mL albumin,rTsP53 protein (6 mg/kg) + 0.1 mL normal saline (NS),IMP (20 mg/kg) + rTsP53 protein (6 mg/kg) respectively, mice in sham group and CLP group were injected intraperitoneally with 0.1 mL albumin + 0.1 mL NS, then these therapies were repeated every 12 hours until the experiment ended. Twenty mice were extracted randomly from each group for surveying 72-hour survival rate.At 0,6,12,24,36,48,72 hours post CLP,3 mice in each group were collected and cytokines in serum were tested by enzyme-linked immunosorbent assay (ELISA).Whole blood was cultured, then the numbers of bacteria colony-forming units (CFU) were counted. Mice were executed at 24 hours, then the epithelial cells ultrastructures of the mice small intestinal mucosa were observed by transmission electron microscope (TEM). ① Compared with CLP,CLP+IMP or CLP+rTsP53 group,72-hour survival rate of the mice in CLP+IMP+rTsP53 group was significantly elevated (85％ vs.20％,55％,25％,all P ＜ 0.05).② No bacteria was found in cultured whole blood of mice in sham group at all time-points. At 6 hours post CLP operation, the number of bacterial clone of all experimental groups was rose significantly. The changed trend of bacterial number in CLP group was rising at the beginning, then declining, and the bacterial number reached the peak at 24 hours (× 106 cfu/L:12.74± 2.33).From 12 hours, the bacterial numbers of CLP+rTsP53 group were higher than those of CLP group, and reached the peak at 36 hours (× 106 cfu/L:22.13 ± 4.28),then declined gradually. The bacterial numbers of CLP+IMP and CLP+IMP+rTsP53 groups reached the peak at 6 hours (× 106 cfu/L:5.72 ± 0.50,5.49 ± 0.59),then declined. They were significantly less than those of CLP group from 12 hours.③ From 6 hours after CLP,the cytokines levels of mice in all experimental groups were higher than those in sham group. The tumor necrosis factor-α (TNF-α) levels in CLP group showed a trend of elevation in the beginning, and decrease thereafter. It reached the peak at 36 hours (ng/L:1 422.67 ±72.19).The TNF-α level peak time of CLP+IMP group,CLP+rTsP53 group,CLP+IMP+rTsP53 group was advanced to 12 hours post CLP (ng/L:1376.29±44.67,929.36±40.42,809.61±22.61).At 24 hours post CLP, the interleukin-6 (IL-6) level of CLP group and CLP+IMP group reached the peak (ng/L:215.39 ± 16.05,191.63 ± 8.99).The peak time of CLP+rTsP53 group and CLP+IMP+rTsP53 was advanced to 12 hours post CLP (ng/L:113.01 ± 12.11,92.43±6.11).The level of IL-4,IL-10 in CLP group raised gradually to the highest at 72 hours (ng/L:366.25 ±24.25,923.14±30.36).The IL-4 and IL-10 levels of CLP+IMP group raised to their maximum value at 12 hours and 24 hours respectively (ng/L:281.47±16.33,555.67±13.57),then declined. The IL-4 and IL-10 levels of CLP+rTsP53 group and CLP+IMP+rTsP53 group gradually ascended their peak value at 72 hours [IL-4 (ng/L) was 453.14±18.53,410.43 ± 15.75,IL-10 (ng/L) was 1 185.61 ± 16.74,1 006.77 ± 36.91,respectively].From 12 hours, the pro-inflammatory cytokines levels of CLP+IMP+rTsP53 group were significantly less than those of CLP+IMP group and CLP+rTsP53 group.④ At 24 hours post CLP, compared with mice in CLP,CLP+IMP, or CLP+rTsP53 group, mice in CLP+IMP+rTsP53 group had slighter ultra structure injuries in the microvilli, cell junction and mitochondria of small intestinal mucosa epithelial cells. The levels of pro-inflammatory cytokines were reduced and the levels of anti-inflammatory eytokines were escalated by intervention of rTsP53 protein combining with IMP boosted in polymierobial septic mice serum, and enhanced the survival rate of the mice. The injection of rTsP53 protein alone had no protective effects on polymicrobial septic mice,because the amount of bacteria in mice blood was augmented

Objective To study the effects of Trichinella spiralis (T.spiralis) infection on mice with oxazolone (OXZ)-induced colitis and the possible immunologic mechanism. Methods Female BALB/c mice at age 6-8 weeks were randomly divided into four groups (A to D). Each mouse in groups B and D was infected with T. spiralis strains. Twenty-one days after T. spiralis infection, the mice in groups A and B were treated with 50% ethanol solution, while those in groups C and D were treated with OXZ to induce the murine model of colitis. Several parameters including survival rate, disease activity index (DAI), macroscopic damage and histological score, myeloperoxidase (MPO) activity and the expression of cytokines (IFN-γ, IL-4 and IL-10) in colonic and splenic tissues at mRNA and protein levels were measured 3 and 7 days after modeling. Results No significant differences in the survival rate, DAI score, macroscopic damage score, histological score, MPO activity were observed between mice from groups C and D (P>0.05). Pre-exposing the mice to T. spiralis strains neither alleviated the mucosal damages nor aggravated the condition of colitis. Compared with group C, the expression of IFN-γ on the third day and the expression of IFN-γ, IL-4 and IL-10 on the seventh day at mRNA and protein levels in colon and spleen tissues were significantly increased in mice treated with T. spiralis and OXZ (P<0.05). The expression of IL-10 at transcriptional level in spleen tissues on the third day was higher than that of group C (P<0.05). The expression of IL-4 and IL-10 at protein level in colon tissues on the third day were significantly up-regulated as compared with those of group C (P<0.05). Conclusion The severity of OXZ-induced colitis in the murine model was neither alleviated nor aggravated by pre-exposing the mice to T. spiralis strains. High doses of IL-10 not only weakened the regulatory effects on Th2 responses, but also induced the production of proinflammatory cytokines, resulting in a new drift of Th1/Th2 without aggravating Th2 responses. Further investigation on the mechanism of T. spiralis-induced over-expression of IL-10 should be conducted, which might increase the practicability of using T. spiralis strains against OXZ-induced colitis. Key words: Trichinella spiralis; Inflammatory bowel disease; Mechanism

So far parasitic disease is still the one of the most important public health problems world-wide,especially in the developing countries.With the development of immunology and molecular biology technology,the parasite vaccine has become one of the important means of controlling parasitic diseases.Immune dose of the parasitic vaccine is one of the important factors for the immune effects.If the dose of vaccine is too low,the body can't be stimulated to produce enough immune response; if the dose of vaccine is too high,it may cause immune tolerance or toxic reactions.Therefore,to determine the appropriate immunization dose is the key of vaccine research.This article reviewed the research status of immune dose of several parasite vaccines,such as Schistosoma,Toxoplasma gondi and Trichinella spiralis. Key words: Parasite ; Vaccine; Immune dose

Objective To predict the antigenicity of two new Trichinella genes,Ad48 h-Ts3 and tetraspanin,synthesize their gene sequences,express their recombinant proteins and evaluate their potential for diagnosis.Methods The potential antigenicity of the two proteins,tetraspanin family protein and adult stage protein Ad48 h-Ts3 were determined according to the result of their hydrophilicity and hydrophobicity predictions.The sequences of these two genes were synthesized after codon optimization and were subcloned into pET28a expression vector and expressed in E.coli system.Western blotting was performed to evaluate the diagnosis value of the proteins using mouse infected sera.Results Bioninformatic analysis showed that tetraspanin gene of Trichinella spiralis,which bears two transmembrane domains,has homology gene in other species.However,Ad48 h-Ts3 gene,which has plenty of antigenic epitopes,is unique in Trichinella spiralis.Genes were subcloned into expression vector and recombinant Ad48 h-Ts3 protein was expressed successfully in E.coli system while no obvious expression was observed in tetraspaning expression.Western blotting showed the expressive recombinant Ad48 hTs3 protein could be recognized by Trichinella infected sera obtained from mice.Conclusion The adult stage protein,Ad48 h-Ts3 protein was expressed successfully in E.coli and had a potential value for diagnosis. Key words: Trichinella spiralis; Diagnosis; Tetraspain; Adult stage protein

Objective To detect Trichinella spiralis DNA by loop-mediated isothermal amplification (LAMP).Methods DNA was extracted by phenol-chloroform extraction method from Trichinella larva.Four primers were designed based on 18S rRNA sequence and used for LAMP assay.To evaluate the specificity of the LAMP method,DNA sample derived from Schistosoma japonicum was used as control.For evaluating the sensitivity of LAMP,the genomic DNA derived from T.spiralis was serially diluted and amplified by LAMP.LAMP reaction mixture was stained with SYBR green I dye and the result was determined according to the color developed in the tube by the naked eye.Green indicated positive reaction while brown was negative.Results After LAMP reaction,the test tube which used genomic DNA derived from T.spiralis appeared to be green,while control was brown.The sensitivity of LAMP assay in this study reached up to 415 fg/μl.Conclusion LAMP assay was established for the detection of DNA sample of T.spiralis. Key words: Trichinella spiralis ; Loop-mediated isothermal amplification ; Detection ;

Trichinosis is one of the most common zoonotic parasitic diseases,a kind of important foodborne parasitic disease with serious harm to human health.Dendritic cells are a kind of innate immune cells.As one of the most important antigen-presenting cells in the intestinal mucosal immune system of the host,it is closely related to the intestinal immune system.In recent years,its effect on parasite infection and host immune responses,especially in the Trichinella spiralis infection and immunity has attracted people's attention.In this review,the research progress at home and abroad on the host intestinal mucosal dendritic cells involved in a series of immunological reactions and roles in Trichinella infection was reviewed. Key words: Trichinella spiralis; Dendritic cells; Immunity; Intestinal mucosa

Objective To analyze the protein components and the immunoreactivity of Trichinella spiralis new born larva (NBL) soluble antigens at different stages in Dali,Yunnan.Methods The sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) was used to analyze protein components of the worm soluble antigens of NBL at different stages, and the Western-blot was used to analyze the immunoreactivity of the worm soluble antigens of NBL at different stages against T.spiralis infected rats.Results The results from SDS-PAGE analysis displayed that the worm soluble antigens of T.spiralis NBL on 24 h showed 36 protein bands with the range between Mr160000 and 10000,among which the 8 main bands were Mr73000,64000,58000,54000,46000,40000,34000 and 25000,respectively.The worm soluble antigens of T.spiralis NBL on 48 h displayed that 40 proteins bands with the range between 172000 and 10000,among which the 17 main bands were 120000,102000,89000,87000,79000,74000,72000,64000,58000,54000,46000,40000,34000,32000,25000,18000 and 10000,respectively.The results of Western-blot showed that 7 protein bands of the worm soluble antigens from T.spiralis NBL on 24 h reacted with the serum of rat anti-T.spiralis NBL.Whereas,14 protein bands of the worm soluble antigens from T.spiralis NBL on 48 h reacted with the serum of rat anti-T.spiralis NBL.Moreover,the negative serum had no cross-reactivity to corresponding proteins.Conclusion The worm soluble antigens of T.spiralis NBL on 24 h had 7 same protein components in comparison with NBL on 48 h.The protein components from the worm antigens of T.spiralis NBL increased during the development of the worm.Western-blot displayed that the serum of rat anti-T.spiralis NBL had stronger specific recognition to the 6 protein components from the worm soluble antigen of T.spiralis NBL both on 24 h and 48 h.Thus,these protein components had stronger immunoreactivity. Key words: Ttichinella spiralis; New born larvae; Worm soluble antigens; Protein components; Immunoreactivity

Out of the 48 Asian countries, Trichinella infection has been documented in domestic animals and in wildlife of 11 (22.92%) and 14 (29.17%) countries, respectively. Human trichinellosis has been reported in 18 (37.5%) countries of Asia. The geographical distribution, history and present situation of epidemic, source of infection and etiological agent of trichinellosis in Asian countries were reviewed. Key words: Trichinella; Trichinellosis; Epidemiology; Asia