Lycium barbarum polysaccharide promotes the immunoprotective effects of a recombinant Lactobacillus plantarum vaccine expressing the Trichinella spiralis cathepsin F-like protease 1 gene.

Trichinellosis caused by Trichinella spiralis (T. spiralis) is a zoonotic disease that poses a substantial risk to human health. At present, vaccines used to prevent trichinellosis are effective, but the production of antibody levels and immunogenicity are low. Adjuvants can increase antibody levels and vaccine immunogenicity. As a result, it is critical to develop an effective adjuvant for the T. spiralis vaccine. Recent research has shown that traditional Chinese medicine polysaccharides with low-toxicity and biodegradability can act as adjuvants in vaccines. In this study, BALB/c mice were orally inoculated with a recombinant Lactobacillus plantarum (L. plantarum) vaccine expressing the T. spiralis cathepsin F-like protease 1 gene (rTs-CPF1), which was given three times at 10-day intervals. Lycium barbarum polysaccharide (LBP) was administered orally for 37 days. At 37 days after the first immunization, mice were infected with 350 T. spiralis muscle larvae (ML). Specific IgG and sIgA antibody levels against the T. spiralis CPF1 protein were increased in mice immunized with rTs-CPF1+LBP compared to those immunized with rTs-CPF1 alone. Furthermore, LBP increased IFN-γ and IL-4 expression levels, and the number of intestinal and intramuscular worms was significantly reduced in the rTs-CPF1+LBP group compared to that in the rTs-CPF1 group. In the rTs-CPF1+LBP group, the reduction rates of adult worms and muscle larvae were 47.31 % and 68.88 %, respectively. To summarize, LBP promotes the immunoprotective effects of the T. spiralis vaccine and may be considered as a novel adjuvant in parasitic vaccines.

Extract of Torreya nucifera Pericarps Exhibits a Parasiticidal Effect on the Nematode Parasite, Trichinella spiralis.

Benzimidazole derivatives can effectively treat nematode parasitic infections; however, some derivatives demand distinct administrative strategies depending on plasma concentration and patient conditions. Numerous studies have examined the potential of natural extracts to exert parasiticidal activity with minimal side effects. Herein, we examined the potential parasiticidal effects of Torreya nucifera extract. The pericarps of T. nucifera were extracted with methanol, dried, and the pellet was dissolved in hot water (Tn-Phw). We designed four individual mouse experiments to clarify the prophylactic and therapeutic effects of Tn-Phw on Trichinella spiralis infection. Also, 100 L1 larvae were isolated and treated with Tn-Phw (10 mg/mL) in vitro to confirm the killing effect. Furthermore, we microscopically examined the morphology of L1 larvae to confirm the parasite-killing effect and analyzed the morphology using a scanning electron microscope (SEM). The expression of three molting-related genes was confirmed to determine whether Tn-Phw induced morphological changes in L1 larvae. Following treatment with Tn-Phw, L1 larvae death was observed after 16 h. Following SEM examination, the healthy muscle larvae showed striated ridges and wrinkles; this was not observed in extract-treated muscle larvae. Expression levels of the three molting-related genes did not differ between the Tn-Phw-treated and control groups. T. spiralis-infected mice pretreated with Tn-Phw showed significantly reduced muscle larva infection when compared with control mice. In all experiments, treatment with Tn-Phw afforded preventive and therapeutic effects against T. spiralis infection and parasitism. Natural substances against nematode parasites could be developed as therapeutic agents with few side effects and enhanced parasiticidal efficacy.

Stem cell biotherapy: A new remedy for Trichinella spiralis-induced inflammatory myopathy.

Trichinella spiralis (T. spiralis)-induced myopathy is an inflammatory myopathy that is difficult to treat unless the parasite is combated in its early intestinal phase before it reaches the muscles. This study aimed to evaluate the effect of local mesenchymal stem cell (MSC) therapy on T. spiralis-induced inflammatory myopathy in rats. Rats were divided into four groups: Group 1 (non-infected non-treated group); Group 2 (infected non-treated group); Group 3 (infected albendazole (ABZ)-treated group); and Group 4 (infected MSC-treated group). Their muscle status was assessed physiologically with the righting reflex and electromyography (EMG), parasitologically with the total muscle larval count, histopathologically with hematoxylin and eosin and Mallory's trichrome stains, as well as immunohistochemically for myogenin as a marker of muscle regeneration. Additionally, serum muscle enzymes creatine kinase (CK) and lactate dehydrogenase (LDH), as well as muscle matrix metalloproteinases MMP1 and MMP9, were assayed. Finally, the immunological response was assessed by measuring the levels of the muscle inflammatory cytokines tumor necrosis factor-alpha (TNF-α), interferon-gamma (INF-γ), and interleukin-4 (IL-4). Our findings revealed that MSC therapy markedly improved muscle EMG and righting reflex, as well as the histopathological appearance of the muscles, reduced inflammatory cellular infiltrates, and increased myogenin immunostaining. It also reduced serum CK and LDH levels, as well as muscle INF-γ, TNF-α, IL-4, MMP1, and MMP9 levels. However, it had no effect on the total muscle larval count. Accordingly, due to its anti-inflammatory properties and muscle-regenerative effect, MSC therapy could be a promising new remedy for T. spiralis-induced myopathy.

Evaluation of the therapeutic effect of Olibanum extract against enteric and intramuscular phases of trichinosis in experimentally infected mice.

Trichinosis is a global food-borne zoonotic disease. Most drugs used in its treatment have low bioavailability and reduced activity against larvae. Therefore, there is an urgent need for safe and effective medications. This study aimed to investigate the in vivo anti-parasitic and anti-inflammatory efficacy of olibanum (OL) extract, alone or combined with albendazole (ABZ) during both intestinal and muscular phases of trichinosis. Male Swiss albino mice (n = 130) were allocated to seven groups, with 20 mice in each group except for the negative control group (10 mice): negative control (GI), positive control (GII), OL25- treated (GIII), OL50- treated (GIV), ABZ50- treated (GV), OL25 + ABZ25 (GVI), and OL50 + ABZ25 (GVII). For intestinal and muscular phase analysis, each group was divided into two subgroups based on euthanizing day (6 and 35 days post-infection). The drug's efficacy was evaluated through parasitological, biochemical, histopathological, and immunohistochemical studies. OL extract at both concentrations (25 mg/kg/d, 50 mg/kg/d) significantly reduced adult (53.7% and 68.1%, respectively) and larval counts (57.3% and 78.8%, respectively). It improved the histopathological changes in intestine and muscle. The expression of CD8+ T cells and the serum level of IL-10 increased significantly during both intestinal and muscular phases (P < 0.05) in OL50 treated mice. Additionally, OL decreased abnormal levels of liver enzymes (ALT & AST). Its effects were dose-dependent in both adult and larval stages. In conclusion, OL exhibits promising in vivo activity against both stages of Trichinella spiralis infection, particularly at the intramuscular phase. It can be safe as an alternative treatment for trichinosis.

Evaluation of Immunotropic Activity of Iridoid-Anthocyanin Extract of Honeysuckle Berries (Lonicera caerulea L.) in the Course of Experimental Trichinellosis in Mice.

Our experiment determined the immunotropic activity of a natural, iridoid-anthocyanin extract from honeysuckle berry (Lonicera caerulea L.) (LC). The extract was administered to mice infected with Trichinella spiralis, orally at a dose of 2 g/kg bw, six times at 24 h intervals (from day 3 prior to the infection to day 3 post-infection (dpi) with T. spiralis. At 5, 7, 14, and 21 dpi, samples of blood, spleen, and mesenteric lymph nodes (MLN) were collected, and isolated lymphocytes were analyzed by flow cytometry. The splenocyte proliferation was estimated with MTT testing, and the intensity of intestinal and muscle infection was also studied. LC stimulated the local immune system by inducing lymphocyte proliferation in the spleen 7 dpi and altered the percentage and absolute count of B (CD19+) and T (CD3+, CD8+) cells 7, 14, and 21 dpi in the peripheral blood. LC extract affected the dynamics of expulsion of adult Trichinella from the intestines and prolonged the intestinal phase of the infection but did not change the number of larvae in the muscles. These results suggest that Lonicera caerulea L. fruit extract modulates murine cellular immune response during intestinal phase of T. spiralis infection but shows no antiparasitic activity.

Effects of probiotic supplementation on post-infectious irritable bowel syndrome in rodent model.

BACKGROUND: Probiotics have been reported to be the active component used in the treatment of many functional gastrointestinal symptoms and syndromes. Lactobacillus and yeast culture are extensively used in probiotic supplements and traditional treatments for irritable bowel syndrome (IBS). The aim of this study was to investigate the effects of probiotic treatments (Lactobacillus acidophilus LA5, Bifidobacterium animalis subsp. lactis BB12 and Saccharomyces cerevisiae var. boulardii) on the behavioral response, targeted gene expression and pro-inflammatory cytokine levels of Pi (Post infectious)-IBS -induced mice. METHODS: Pathogen-free male C57L/B6 mice and the Trichinella-infected mice were used to measure the score of abdominal withdrawal reflex (AWR). To compare molecular, biological and biochemical evidences of given probiotics with normal and positive control groups in mice, we conducted quantitative reverse transcription polymerase chain reaction (RT-qPCR), western blotting, and cytokine analysis. RESULTS: Pi-IBS-induced immune response was confirmed that PAR-2 mRNA level was significantly increased by Trichinella infection (P < 0.05). The reduction of Pi-IBS symptoms through Trichinella infection and the effects of given probiotics were confirmed by a change in the protein levels of cytokines (P < 0.05). In addition, the administration of DW (Daewon) probiotics significantly decreased serum levels of IL-1 and IL-6 (P < 0.05). CONCLUSIONS: We have demonstrated that the given probiotics decreased pro-inflammatory cytokine levels in both the control and Pi-IBS induced mice. Taken all the results together, the results support that DW probiotics has a potential as a probiotic medication for patient with IBS via regulating TNF-α and IL-6 protein levels and serum IL-1 and IL-6 levels.

Vaccaria n-Butanol Extract Lower the Production of Proinflammatory Cytokines and the Infection Risk of T. spiralis In Vivo.

INTRODUCTION: Trichinellosis is a severe zoonosis involving the activation of inflammatory cells, accompanied by the prominent expressions of proinflammatory cytokines in the host. Semen vaccariae, the seeds of Vaccaria segetalis (Neck.) Garcke. ex Asch. (Caryophyllaceae), is a famous traditional herb that is rich in vaccaria n-butanol extract (VNE). Vaccarin is one major active component of VNE, and it is reported in the treatment of stranguria disease. Hypaphorine is another main active component of VNE and has good anti-inflammatory effect, whereas the potential bioactivity of VNE in trichinellosis treatment is still unknown. MATERIALS AND METHODS: This study was designed to evaluate the potential anthelmintic and anti-inflammatory activity of VNE toward T. spiralis infection. ICR mice were used to assess the effect of VNE on repression larvae and adult worms in vivo. Immunohistochemistry analysis was performed to evaluate the expression levels of IL-1ß, IL-6, TNF-α, and COX-2. RESULTS: Our results showed that VNE could effectively depress the expressions of proinflammatory cytokines, including IL-1ß, IL-6, TNF-α, and COX-2. The adult worms were decreased by 79.53%, while the muscle larvae were diminished by 77.70% as compared to the control. CONCLUSION: These results demonstrated that VNE may be a promising therapeutic agent against the inflammation and diseases caused by T. spiralis infection.

Taurine drinking attenuates the burden of intestinal adult worms and muscle larvae in mice with Trichinella spiralis infection.

The parasitic nematode Trichinella spiralis can cause trichinellosis, which leads to pathological processes in the intestine and muscle. The intestinal invasion determines the development, subsequent course, and consequences of the disease. Gastrointestinal nematode infection, including with T. spiralis, is accompanied by a rapid and reversible expansion of mucosal mast cell and goblet cell in the intestinal epithelium, which play important roles in the host immune response to parasite and worm expulsion from the intestine. Taurine and its derivatives have anti-infection and anti-inflammatory properties. We investigated whether taurine supplementation in mice could influence the development and pathological processes of infection with T. spiralis. Supplementing 1% taurine in drinking water in mice infected with T. spiralis could alleviate the burden of intestinal adult worms on days 7 and 10 postinfection (all p < 0.01) and the formation of infective muscle larvae in striated muscle during T. spiralis infection (p < 0.01). As compared with T. spiralis infection alone, taurine treatment increased the number of goblet cells on days 7, 10, and 15 (p < 0.01 and p < 0.05) and alleviated intestinal mucosal mast cell hyperplasia on days 10 and 15 (all p < 0.01). So taurine supplementation in drinking water increased infection-induced intestinal goblet cell hyperplasia and ameliorated mucosal mastocytosis. Thus, taurine can ameliorate the pathological processes of trichinellosis and may be of great value for the treatment and prevention of infection with T. spiralis and other gastrointestinal nematodes.

Calcification of Trichinella spiralis larval capsule.

Trichinella spiralis larvae were examined in TEM to identify calcareous corpuscles in the outer part of parasite capsule. The microroentgenographic analysis of calcareous corpuscles mainly demonstrated the presence of phosphorus and calcium. The physiological importance of calcareous corpuscles, as well as their significance in decay of T. spiralis larvae were discussed.

Cloning and analysis of a novel cDNA from Trichinella spiralis encoding a protein with an FYVE zinc finger domain.

A cDNA library from Trichinella spiralis adults 3 days post-infection was screened with a cDNA probe, designated T 54, derived from a newborn larvae subtracted cDNA library. Sequence analysis showed that the positive clone contained a cDNA insert of 1464 bp in length with a single open reading frame of 1290 bp, which encoded a protein of 429 amino acids with a putative molecular mass of 49.9 k Da. Database analysis predicted the deduced protein had a leucine zipper motif and an FYVE zinc finger domain. The recombinant fusion protein was expressed and rabbit anti-recombinant protein sera reacted with a single peptide migrating at approximately 55 k Da in crude worm extract from muscle larvae, adults and newborn larvae stages.

Polyunsaturated fatty acids and parasitism: effect of a diet supplemented with fish oil on the course of rat trichinellosis.

Dietary fish oil has a beneficial effect on heart and some bacterial diseases and apart from other effects, some studies have revealed their ability to modulate the course of inflammatory and autoimmune diseases. The study here reported was designed to evaluate the possible influence of a fish oil supplement on the course of a Trichinella infection. Nutritional, parasitological and immunological parameters were analyzed. Two groups of 20 Wistar rats, one fed a standard diet and the other one a standard diet supplemented with fish oil, were infected with 1000 L1 larvae. Other two uninfected groups served as control. Results were as follows: fish oil diet intake and infection have, respectively, a positive and a negative effect on growth and food utilization. The negative effect is detected later in animals fed the fish oil diet. A reduction of 30.9 and 36.6% in the number of adult worms and L1 larvae, respectively, was observed in the fish oil group as compared to the standard diet group. Production of IFNgamma (Th1 response) and IL4 (Th2) response was measured in stimulated splenic cells. The fish oil diet increased both IFNgamma and IL4 levels. At 6 days after infection both IFNgamma and IL4 responses were detected, but at 36 days after infection only IL4 was detected in the standard group. The level of somatic and cuticular antibodies was not affected by the diet.

Trichinella pseudospiralis: X-ray analysis of the infected muscle cells and the muscle larva compartments after exponential growth of the larva in situ.

The distribution of nutrients such as Na, Cl, K, P, S, and Ca was investigated by energy-dispersive X-ray microanalysis (EDXA) on bulk-frozen mouse muscles infected with Trichinella pseudospiralis. In an attempt to gain information as to whether muscle larvae would modify the element status within muscles of their host. Significant increases in phosphorus concentrations within nurse cells and internal cells of the larva, e.g., the somatic muscle cells and the stichocytes, were shown. This could reflect changes in the energy metabolism and/or in the turnover of nucleic acids of the nurse cell. High phosphorus concentrations within the internal larva cells corresponded to high levels of expression of thymidylate synthase shown elsewhere in both T. pseudospiralis and T. spiralis muscle larvae, most likely reflecting more intensive DNA replication. Furthermore, the Na+-concentration gradient between the nurse cell and neighboring host muscle cells indicated the plasma membrane as a diffusion barrier for Na+. Moreover, the cuticle seemed to maintain the Cl--concentration gradient between the nurse cell and the internal cells of the larva. Nevertheless, the mechanisms involved in the transport of both ions, the collagen cuticle appeared, strong permeability that serves the transcuticular transport maintaining the Cl--concentration gradient.

[Changes in the invasive process and the immune response of the host during the mebendazole and voltaren treatment of experimental trichinelliasis in guinea pigs]. / Izmenenie invazionnogo protsessa i immunnogo otveta khoziaina pri lechenii mebendazolom i vol'tarenom éksperimental'nogo trikhinelleza morskikh svinok.

Guinea pigs infested with Trichinella (5-10 larvae per 1 g body weight) at the intestinal (at days 2-4 of infection), migration al (at days 6-11), and muscular (at days 22-27) stages of invasion, were given voltaren, 2.14 mg/kg daily, and mebendazole, 75 mg/kg daily. The amount of adult Trichinella in small intestine and larvae in the diaphragm, T- and B-lymphocyte count (T-REC and B-RFC), functional leukocyte activity in RTML for FGA and Trichinella larvae antigens, content of circulating immune complexes and complement were determined. Voltaren did not cause delay in adult Trichinella elimination from the intestine of experimental animals, it also did not increase invasion rate at the muscular stage. As compared to the control, T- and B-lymphocyte content and RTML for FGA remained unaltered. Experimental animals show elevated blood complement activity and RTML inhibition of trichinella antigens. The combination of mebendasole and voltaren did not reduce parasitogenic effect. Despite the terms of drug injection, T- and B-lymphocytes count and complement levels in blood were found to increase, as well as the RTML index on FGA and its inhibition of specific antigens as compared with the infected untreated animals.

Rapid expulsion of Trichinella spiralis in suckling rats.

Orally administered Trichinella spiralis muscle larvae were rapidly expelled by rat pups suckling an immune dam. The immunity was delivered in the milk; substantial resistance was conferred on normal rat pups suckled for only 24 hours by a Trichinella-immune foster mother. The pups were protected by oral or systemic administration of specific serum antibodies. When infused into a normal lactating dam, these antibodies accumulated in the serum of her suckling pups.