Trichinella spiralis inhibits myoblast differentiation by targeting SQSTM1/p62 with a secreted E3 ubiquitin ligase.

Trichinella spiralis infection is associated with the formation of cysts within host skeletal muscle cells, thereby enabling immune evasion and subsequent growth and development; however, the pathogenic factors involved in this process and their mechanisms remain elusive. Here, we found that Ts-RNF secreted by T. spiralis is required for its growth and development in host cells. Further study revealed that Ts-RNF functions as an E3 ubiquitin ligase that targets the UBA domain of SQSTM1/p62 by forming K63-type ubiquitin chains. This modification interferes with autophagic flux, leading to impaired mitochondrial clearance and abnormal myotube differentiation and fusion. Our results established that T. spiralis increases its escape by interfering with host autophagy via the secretion of an E3 ubiquitin ligase.

Inactivation of Toxoplasma gondii in dry sausage and processed pork, and quantification of the pathogen in pig tissues prior to production.

Toxoplasma gondii is an important zoonotic foodborne parasite. Meat of infected animals appears to be a major source of infection in Europe. Pork is the most consumed meat in France, with dry sausages well represented. The risk of transmission via consumption of processed pork products is largely unknown, mainly since processing will affect viability but may not entirely inactivate all T. gondii parasites. We investigated the presence and concentration of T. gondii DNA in the shoulder, breast, ham, and heart of pigs orally inoculated with 1000 oocysts (n = 3) or tissue cysts (n = 3) and naturally infected pigs (n = 2), by means of magnetic capture qPCR (MC-qPCR). Muscle tissues of experimentally infected pigs were further used to evaluate the impact of manufacturing processes of dry sausages, including different concentrations of nitrates (0, 60, 120, 200 ppm), nitrites (0, 60, 120 ppm), and NaCl (0, 20, 26 g/kg), ripening (2 days at 16-24 °C) and drying (up to 30 days at 13 °C), by a combination of mouse bioassay, qPCR and MC-qPCR. DNA of T. gondii was detected in all eight pigs, including in 41.7% (10/24) of muscle samples (shoulder, breast and ham) and 87.5% (7/8) of hearts by MC-qPCR. The number of parasites per gram of tissue was estimated to be the lowest in the hams (arithmetic mean (M) = 1, standard deviation (SD) = 2) and the highest in the hearts (M = 147, SD = 233). However, the T. gondii burden estimates varied on the individual animal level, the tissue tested and the parasitic stage used for the experimental infection (oocysts or tissue cysts). Of dry sausages and processed pork, 94.4% (51/54) were positive for T. gondii by MC-qPCR or qPCR, with the mean T. gondii burden estimate equivalent to 31 parasites per gram (SD = 93). Only the untreated processed pork sample collected on the day of production was positive by mouse bioassay. The results suggest an uneven distribution of T. gondii in the tissues examined, and possibly an absence or a concentration below the detection limit in some of them. Moreover, the processing of dry sausages and processed pork with NaCl, nitrates, and nitrites has an impact on the viability of T. gondii from the first day of production. Results are valuable input for future risk assessments aiming to estimate the relative contribution of different sources of T. gondii human infections.

One Health: A social science discussion of a global agenda.

This article introduces the Parasite issue dedicated to part of the research in social sciences supported by the Domaine d'Intérêt Majeur de la Région Île-de-France (DIM) One Health [2016-2022]. We show how the four papers of this special issue are related. Jérôme Michalon recalls the genealogy of One Health and analyzes it as an "epistemic watchword". Using antibiotic resistance as a case study, Estera Badau demonstrates how "One Health" results from a series of formulas and the bringing together of a plurality of fields and actors. Nicolas Lainé and Serge Morand show how One Health fits in with attempts already initiated in the colonial period and context. They highlight the need to (re)legitimize local and non-human knowledge, in order to truly decolonize One Health and better prevent epidemic emergence. Finally, Frédéric Keck, Nicolas Lainé, Arnaud Morvan and Sandrine Ruhlmann show how zoonotic reservoir and cultural practices are linked in the context of three specific societies. This paper highlights two main contributions of social sciences: 1) To think about One Health genealogy, how the question is framed and by which actors. The questions of practices, social representations but also of the environment are less present than the issues of human and animal medicine. The Anthropocene, the Capitalocene, even some of its variations such as the "domesticoscene" thus appear to be key elements. 2) To propose methods and tools that make One Health operational, advocating a less asymmetrical view of types of knowledge (scientific, local, non-human) and more contextualized global health recommendations.

Title: Une seule santé : discussion en sciences sociales d'un programme mondial. Abstract: Cet article introduit le numéro spécial de la revue Parasite dédié à des travaux soutenus par le Domaine d'Intérêt Majeur de la Région Île-de-France One Health [2016­2022]. Nous montrons ainsi l'articulation entre les quatre articles constituant ce numéro. Jérôme Michalon rappelle la généalogie de One Health et propose une définition qui engage : un mot d'ordre épistémique. Estera Badau illustre le propos avec le cas de l'antibiorésistance en montrant comment One Health résulte d'une série de formulations et de la mise en relation d'une pluralité de domaines et d'acteurs, notamment scientifiques, gestionnaires ou porteurs de politiques publiques. Nicolas Lainé et Serge Morand montrent la façon dont One Health s'inscrit dans des tentatives déjà initiées en période et contexte colonial. Dans ce cadre, ils mettent en avant la nécessité de (re)légitimer le savoir local et celui des non-humains, afin de réellement décoloniser One Health et de mieux prévenir les émergences épidémiques. Enfin, Frédéric Keck, Nicolas Lainé, Arnaud Morvan et Sandrine Ruhlmann montrent comment réservoir zoonotiques et pratiques culturelles s'articulent dans le cadre de trois sociétés spécifiques. Cet article met ensuite en avant deux apports principaux des sciences sociales. 1) réfléchir à la généalogie de One Health, à la façon dont la question est cadrée et par quels acteurs. Il apparait ainsi que les questions de pratiques, de représentations sociales mais aussi d'environnements sont moins présentes que les enjeux de médecine humaine et animale, alors même que les zoonoses sont fortement liées à des modifications des relations entre humains, animaux et environnement. L'anthropocène, et sans-doute plutôt le capitalocène, voire certaines de ses déclinaisons, comme le « domesticoscène ¼ apparaissent ainsi comme des éléments clés. 2) Proposer des méthodes et des outils qui permettent de rendre opérationnel One Health, en plaidant pour une vision moins asymétrique des types de savoirs (scientifiques, locaux, non-humains) et plus contextualisée des recommandations de santé globale.

Spatial and Temporal Circulation of Babesia caballi and Theileria equi in France Based on Seven Years of Serological Data.

Caused by two blood parasites, Babesia caballi and Theileria equi, equine piroplasmosis is a tick-borne disease that poses major health and economic issues for the equine industry. Our objective was to gain insight into the spatio-temporal variations of parasite circulation in France, where the disease is known to be enzootic, but has been the subject of few studies. Seroprevalence was assessed for each parasite thanks to 16,127 equine sera obtained between 1997 and 2003 from all over France and analysed through complement fixation tests. Results indicated that 13.2% (5-27% depending on the region) of horses were seropositive for T. equi and 9.5% (3-25%) for B. caballi. Regardless of the year, horses from the southern regions of France were the most affected by B. caballi or T. equi infection, while the proportion of horses having antibodies against T. equi increased over time. These results highlight the heterogeneity of the circulation of both piroplasms, which may be linked with ecological diversity and vector distribution. Our data provide baseline information regarding the sero-epidemiology of B. caballi and T. equi infection in horses in France, making it now possible to select regions for future studies on risk factors, and design and implement effective targeted measures against equine piroplasms.

The Anti-Inflammatory Immune Response in Early Trichinella spiralis Intestinal Infection Depends on Serine Protease Inhibitor-Mediated Alternative Activation of Macrophages.

Trichinella spiralis is recognized for its ability to regulate host immune responses via excretory/secretory (ES) products. Serine protease inhibitors (serpins) play an important role in ES product-mediated immunoregulatory effects during T. spiralis infection. In this study, the immunoregulatory properties of a serpin derived from T. spiralis (Ts-serpin) were explored in BALB/c mice. The results showed that naturally occurring Ts-serpin was detected in the stichosomes of muscle larvae and adult worms. Moreover, enhancing (by injection of a soluble-expressed recombinant Ts-serpin [rTs-serpin]) or blocking (by passive immunization with anti-rTs-serpin serum) the effects of Ts-serpin changed the levels of cytokines related to inflammation induced by T. spiralis infection in the serum, mesenteric lymph nodes, and peritoneal cavity, which then led to a change in the adult worm burden in early T. spiralis infection. Moreover, the phenotypic changes in peritoneal macrophages were found to be related to Ts-serpin-mediated immunoregulation. Furthermore, a STAT6 activation mechanism independent of IL-4Rα has been found to regulate protein-mediated alternative activation of bone marrow-derived macrophages and mimic the immunoregulatory role of Ts-serpin in T. spiralis infection. Finally, the anti-inflammatory properties of rTs-serpin and bone marrow-derived macrophage alternative activation by rTs-serpin were demonstrated using a trinitrobenzene sulfonic acid-induced inflammatory bowel disease model. In summary, a protein-triggered anti-inflammatory mechanism was found to favor the survival of T. spiralis in the early stage of infection and help to elucidate the immunoregulatory effects of T. spiralis on the host immune response.

Nod-like receptor pyrin domain containing 3 plays a key role in the development of Th2 cell-mediated host defenses against Trichinella spiralis infection.

The inflammasome is a key line of immune defense against invading infectious pathogens. However, knowledge of the role of nod-like receptor pyrin domain containing 3 (NLRP3) in Trichinella spiralis infection which characteristically induces T-helper 2 (Th2) immune responses is sparse. In this study, we investigated the role of NLRP3 in the protection against T. spiralis infection through the Th2 immune response. We show that NLRP3 expression in CD4+ T cells was significantly increased at 7 days post-infection of T. spiralis. Compared to wild-type (WT) CD4+ T cells, the expression of IL-4 mRNA was reduced in NLRP3-/- CD4+ T cells, however, the expression of IFN-γ mRNA was comparable between the two groups. Consistently, ELISA and flow cytometry analysis showed that NLRP3-/- CD4+ T cells secreted lower levels of IL-4 than CD4+ T cells from WT mice, whilst the levels of IFN-γ secreted by NLRP3-/- CD4+ T cells were of similar levels to those secreted by WT CD4+ T cells. In addition, we observed a significant reduction of IL-4 and IL-13 by ELISA in NLRP3 -/- mice at 1, 2 and 4 weeks post-infection. Furthermore, we found that adult worm survival was substantially prolonged and muscle larvae burden was significantly increased in NLRP3 -/- mice. We further show that NLRP3 promotes the host defense against T. spiralis through its participation in the differentiation of Th2 cells. These findings provide novel insights into parasite expulsion and highlight the importance of NLRP3 in the host defense against T. spiralis.

Comparative analysis of excretory-secretory products of muscle larvae of three isolates of Trichinella pseudospiralis by the iTRAQ method.

Trichinella pseudospiralis is a non-encapsulated intracellular parasitic nematode that can possess a strong ability to modulate the host immune response. Here, we compared the differentially expressed proteins of excretory-secretory (ES) products in three isolates of T. pseudospiralis muscle larvae (ML) [from Russia (RUS), United States of America (USA) and Australia (AUS)] using isobaric tags for relative and absolute quantification (iTRAQ)-based technology. A total of 2591 nonredundant proteins were identified, of which 65 (146), 72 (98) and 43 (103) significantly upregulated (downregulated) differentially expressed proteins were detected among pairwise comparisons (T4RUS vs T4USA, T4AUS vs T4USA and T4RUS vs T4AUS). In addition, GO annotation, KEGG and STRING analyses were carried out on the screened differentially altered proteins. The main biological processes involved included carbohydrate metabolic processes, DNA metabolic processes, cellular protein modification processes and homeostatic processes. The majority of KEGG pathways were found to be related to the metabolic pathways, lysosome pathway and protein processing in endoplasmic reticulum. Moreover, all ES protein expression levels involved in the lysosome pathway were significantly higher in the T4USA isolate than in the other two isolates. We also found differences in the expression of some important immunoregulatory proteins, such as protein disulfide-isomerase, thioredoxin protein and deoxyribonuclease-2-alpha, between different isolates of T. pseudospiralis ML. Flow cytometry was used to detect the increase in the CD4+/CD8 + T-cell ratio in pig peripheral blood and to verify the effect of T. pseudospiralis on the Th1/Th2 polarization of the host. Quantitative real-time PCR analysis also confirmed that the changes in the transcriptional level of genes were consistent with those at the proteomic level. These findings reveal the possible role of significantly differentially expressed proteins in ES products of the different isolates of T. pseudospiralis in antagonizing and participating in the regulation of the host immune response and maintaining a stable growth environment.

Extracellular vesicles derived from Trichinella spiralis prevent colitis by inhibiting M1 macrophage polarization.

Extracellular vesicles (EVs) are membranous containers released by cells that are powerful agents of intercellular communication. EVs have been described for various parasites and are associated with tissue inflammation. Several studies have demonstrated that parasite EVs can have either pro- or anti-inflammatory impacts, depending on the type of parasite. To evaluate the immunomodulatory properties of EVs produced by Trichinella spiralis (T. spiralis), we established a mouse model with dextran sulphate sodium (DSS)-induced colitis. The muscle larvae of T. spiralis were cultured in vitro and the released EVs were isolated by ultracentrifugation. T. spiralis EVs (Ts-EVs) were characterized according to morphology, size and constituent surface proteins (CD63, Enolase and Hsp70). Mice were treated with water containing 3% DSS after last intraperitoneal injection of Ts-EVs. Disease activity index (DAI), macroscopic and histopathological scores of Ts-EVs group was lower than DSS group. And Ts-EVs prevented the increase in the expression of TNF-α, IFN-γ, IL-17A and IL-1ß observed in the colon of DSS-treated mice. In contrast, upregulation of IL-4, IL-10, TGF-ß and IL-13 expression was detected in Ts-EVs+DSS group. In addition, Ts-EVs increased the infiltration of alternatively activated (M2) macrophages into the colon. The expression of CD206 (M2 marker) in the mesenteric lymph nodes (MLN) of mice with colitis increased in Ts-EVs+DSS group. Furthermore, Ts-EVs interfered with both the NF-κB and MAPK signalling pathways. Taken together, our findings demonstrate that Ts-EVs can affect the development of inflammation in DSS-induced colitis by inhibiting M1 macrophage polarization, due to their immunomodulatory ability.

NLRP3 played a role in Trichinella spiralis-triggered Th2 and regulatory T cells response.

Trichinella spiralis maintains chronic infections within its host. Muscle larvae excretory-secretory products (MLES) typically induce parasite-specific immune responses such as the Th2 response and regulatory T cells (Tregs) by modulating dendritic cell (DC) phenotype via the recognition of pattern recognition receptors (PRRs), such as Nod-like receptors (NLRs). We aimed to investigate the role of NLRP3 in T. spiralis-triggered immune response. We found that larvae burden was increased in NLRP3-/- mice compared to wild type (WT) mice. Administration of MLES induced higher levels of IL-4, IL-10, TGF-ß and population of Tregs in WT mice than in NLRP3-/- mice. In vitro, we showed that increased expression of CD40 on the surface of MLES-treated DCs was inhibited after NLRP3 knockout. Increased production of IL-1ß, IL-18, IL-10 and TGF-ß, but not IL-12p70, was significantly diminished in the absence of NLRP3. Furthermore, our results demonstrated that MLES-treated DCs induced higher levels of IL-4, IL-10 and TGF-ß and populations of Tregs in vitro. These inductions were abolished by NLRP3 deficiency in DCs, suggesting that NLRP3 in MLES-treated DCs plays a role in promoting the Th2 and Treg response. Taken together, we identified for the first time the involvement of NLRP3 in host defences against T. spiralis.

Toxoplasma gondii in beef consumed in France: regional variation in seroprevalence and parasite isolation.

In France, the consumption of cattle and sheep meat appears to be a risk factor for infection of pregnant women with Toxoplasma gondii. Several nation-wide surveys in France have investigated the prevalence of T. gondii in sheep and pig meat, but little is known at present about the prevalence of the parasite in beef. The main objective of the present cross-sectional survey was to estimate the seroprevalence of T. gondii infection in beef consumed in France. A secondary objective was to attempt to isolate T. gondii from cattle tissues and to study the geographical and age variations of this seroprevalence. The overall estimate of seroprevalence of T. gondii in bovine carcasses (n = 2912), for a threshold of 1:6 was 17.38%. A strong age effect was observed (p < 0.0001) with a seroprevalence of 5.34% for calves (<8 months) and 23.12% for adults (>8 months). Seroprevalence estimates given by area of birth and area of slaughtering for adults showed that the areas with the highest seroprevalence were not the same between these two variables. Only two strains, corresponding to genotype II, were isolated from heart samples, indicating that there is a limited risk of human infection with T. gondii, which needs to be correlated with the food habit of consuming raw or undercook (bleu or saignant) beef. However, new questions have emerged, especially concerning the isolation of parasites from beef and the precise role of bovines, generally described as poor hosts for T. gondii, in human infection.

TITLE: Toxoplasma gondii dans la viande bovine consommée en France : variation régionale de la séroprévalence et isolement de parasites. ABSTRACT: En France, la consommation de viande bovine et ovine apparaît comme un facteur de risque pour la contamination des femmes enceintes par Toxoplasma gondii. Plusieurs enquêtes nationales ont été réalisées afin de déterminer le niveau de contamination par T. gondii de la viande ovine et porcine, en France, mais très peu est encore connu quant à la prévalence du parasite dans la viande bovine. La présente enquête transversale avait pour objectif principal d'estimer la séroprévalence de l'infection à T. gondii dans la viande bovine consommée en France, ainsi que d'isoler T. gondii à partir de tissus de bovins et d'étudier, à titre d'objectif secondaire, les variations géographiques et d'âge de cette prévalence. L'estimation globale de la séroprévalence de T. gondii dans les carcasses de bovins (n = 2912) était de 17,38 % (pour un seuil de dilution à 1:6). Un effet significatif de l'âge a été observé (p < 0,0001) avec une séroprévalence de 5,34 % pour les veaux (<8 mois) et de 23,12 % pour les adultes (>8 mois). Les estimations de séroprévalence données par zone de naissance et par zone d'abattage pour les adultes montrent que les zones de séroprévalence les plus élevées n'étaient pas les mêmes pour ces deux variables. Seulement deux souches, de génotype II, ont été isolées à partir d'échantillons de cœurs, soulignant que le risque d'infection humaine est limité, mais doit être corrélé avec les habitudes de consommation alimentaire de la viande bovine peu/pas cuite (bleu ou saignante). Cependant, de nouvelles questions se posent, notamment en ce qui concerne l'isolement du parasite à partir de la viande bovine, ainsi que le rôle précis des bovins, généralement décrits comme des hôtes médiocres pour T. gondii, dans la contamination humaine.

Epidemiological surveillance of schistosomiasis outbreak in Corsica (France): Are animal reservoir hosts implicated in local transmission?

Environmental and anthropogenic changes are expected to promote emergence and spread of pathogens worldwide. Since 2013, human urogenital schistosomiasis is established in Corsica island (France). Schistosomiasis is a parasitic disease affecting both humans and animals. The parasite involved in the Corsican outbreak is a hybrid form between Schistosoma haematobium, a human parasite, and Schistosoma bovis, a livestock parasite. S. bovis has been detected in Corsican livestock few decades ago raising the questions whether hybridization occurred in Corsica and if animals could behave as a reservoir for the recently established parasite lineage. The latter hypothesis has huge epidemiological outcomes since the emergence of a zoonotic lineage of schistosomes would be considerably harder to control and eradicate the disease locally and definitively needs to be verified. In this study we combined a sero-epidemiological survey on ruminants and a rodent trapping campaign to check whether schistosomes could shift on vertebrate hosts other than humans. A total of 3,519 domesticated animals (1,147 cattle; 671 goats and 1,701 sheep) from 160 farms established in 14 municipalities were sampled. From these 3,519 screened animals, 17 were found to be serologically positive but were ultimately considered as false positive after complementary analyses. Additionally, our 7-day extensive rodent trapping (i.e. 1,949 traps placed) resulted in the capture of a total of 34 rats (Rattus rattus) and 4 mice (Mus musculus). Despite the low number of rodents captured, molecular diagnostic tests showed that two of them have been found to be infected by schistosomes. Given the low abundance of rodents and the low parasitic prevalence and intensity among rodents, it is unlikely that neither rats nor ruminants play a significant role in the maintenance of schistosomiasis outbreak in Corsica. Finally, the most likely hypothesis is that local people initially infected in 2013 re-contaminated the river during subsequent summers, however we cannot definitively rule out the possibility of an animal species acting as reservoir host.

International Commission on Trichinellosis: Recommendations for quality assurance in digestion testing programs for Trichinella.

Effective performance of digestion testing methods for Trichinella, and their use for the detection of infected animals and the prevention of human trichinellosis require system-wide incorporation of appropriate quality assurance (QA) practices. The recommendations of the International Commission on Trichinellosis (ICT) aim to facilitate reliable test results when laboratories operate within a quality management system (QMS) which includes: 1) a quality manual (or similar documentation of the QMS); 2) a validated test method with identified critical control points; 3) a training program; 4) procedures utilizing proficiency testing and other methods to confirm technical capability of analysts; 5) equipment calibration and maintenance; 6) standard operating procedures, related documentation and reporting; 7) procedures to enable continuous monitoring and improvements; and 8) regular internal and third party audits. The quality manual or similar documentation describes the QMS within a testing laboratory, and lists the QA policies and good laboratory practices. Quality assurance goals contained in such documentation are the foundation of an effective QA program and must be explicit, measurable, and expressed in terms of performance criteria for the test method based on purpose for testing. The digestion method is capable of consistently detecting Trichinella larvae in meat at a level of sensitivity that is recognized to be effective for use in controlling animal infection and preventing human disease. However, consistent performance of the assay is assured only when parameters of the test method have been defined, scientifically validated as fit for purpose, and used within an effective QMS. The essential components of a digestion assay, specifically the critical control points and minimum standards for test performance are described. Reliable proficiency samples and their appropriate use in a quality system are key factors for certifying and maintaining an effective testing laboratory, including qualifying, re-qualifying and disqualifying of analysts as appropriate. Thus recommendations are included for the preparation and use of proficiency samples in a Trichinella digestion testing laboratory. The minimum training requirements for analysts performing a quality assured digestion assay, as well as suggested requirements for the content of a training manual, are also outlined. Finally, these ICT recommendations include essential components and minimum standards for maintaining and achieving certification and maintenance of a laboratory performing digestion testing for Trichinella. The certification program for the laboratory, including qualifying analysts, may be administered by a National Reference Laboratory or an authorized third party certifying body, under the auspices of the appropriate competent authority.

Multilocus genotype analysis outlines distinct histories for Trichinella britovi in the neighboring Mediterranean islands of Corsica and Sardinia.

BACKGROUND: The zoonotic nematode Trichinella britovi was discovered in two neighboring Mediterranean islands of Corsica and Sardinia, almost simultaneously at the beginning of the 21st century. An epidemiological link between the two parasite populations was generally assumed. In 2015, an outbreak of trichinellosis in Nice, the South of France, was reportedly caused by the consumption of raw pork delicatessen imported from Corsica. The aims of the present study were to investigate, by multilocus genotype (MLG) analyses, the hypothesis of the common origin of the Corsican and Sardinian T. britovi foci and to trace "from fork to farm" the origin of the pork product, which caused a trichinellosis outbreak in mainland France in 2015. METHODS: Sixty-three T. britovi isolates were collected from animals and pork products of Sardinia and Corsica islands and from mainland of Italy, France and Spain. We analyzed genetic variability at four polymorphic microsatellite loci by two independent algorithms, the Bayesian and multivariate analyses, to evaluate the genetic relationships of 1367 single larvae. RESULTS: Trichinella britovi isolates of the two islands showed different genetic structures and the Bayesian analysis revealed a different membership of the two insular populations. Furthermore, two geographically separate genetic groups were identified among Corsican isolates. Lastly, the origin of the pork delicatessen marketed in Nice was linked to a breeder-butcher in Corsica. CONCLUSIONS: The low level of genetic admixture of the insular T. britovi isolates suggests that this pathogen colonized the two islands by separate events. On the other hand in Corsica, although the isolates share the same genetic structure, geographically separate isolates showed different membership. We suggest the MLG analysis as a suitable method in supporting epidemiological investigations to trace "from fork to farm" insular populations of T. britovi.

Detection of natural Trichinella murrelli and Trichinella spiralis infections in horses by routine post-slaughter food safety testing.

Trichinella spiralis typically infects domestic swine, wild boar and occasionally horses, has a cosmopolitan distribution, and consequently is most frequently associated with food-borne outbreaks of trichinellosis in humans. Trichinella murrelli is typically found in wild carnivores in temperate areas of North America, where it has been responsible for outbreaks of human trichinellosis due to consumption of infected wild game. There has previously been only indirect evidence of natural infection with T. murrelli in a horse originating from Connecticut and implicated in an outbreak of trichinellosis in France in 1985. We describe a T. murrelli infection detected during routine testing of a horse from the USA imported to Canada for slaughter and export to the European Union (EU). Approximately 5 or more larvae per gram were recovered from digested tongue and diaphragm samples and identified as T. murrelli by PCR. This case provides the first direct evidence of naturally acquired T. murrelli infection in a horse, and further supports the potential food safety risk posed by this parasite species. It is the first instance in Canada of the detection of a Trichinella-infected horse via routine post-mortem testing. Trichinella spiralis-infected horses have been similarly detected by regulatory testing in France, and further details of two such previously reported cases are also provided here. The cases described herein underscore the importance of continued vigilance in quality assured food safety testing of horse meat to mitigate the risk of human trichinellosis.

Trichinella spiralis and Tumors: Cause, Coincidence or Treatment?

BACKGROUND: Conventional therapeutic strategies for tumors have had limited success, and innovative and more effective approaches to treatment are urgently required. The ancient idea that various biological, bacterial, yeast, viral, and parasitic agents can be used as cancer therapeutics has gradually attracted considerable interest. Certain parasites have been widely discussed in association with human and animal tumors. The purpose of this review was to examine previous literatures which investigates the relations between Trichinella spiralis (T. spiralis) and tumors. METHODS: Using PubMed, articles published before 2018 in the whole world have been searched and comprehensively reviewed. RESULTS: Many researches have provided proofs that T. spiralis possesses antitumor activities. The antitumor effect of T. spiralis was first described in the 1970s. However, its research has been inconsistent, and little progress has been made in this field. Therefore, the mechanisms underlying these inhibitory effects are still unclear, and convincing evidence of the links between T. spiralis and the prevention or treatment of tumors from clinical trials is absent. Meanwhile, some other researches also suggested that T. spiralis may cause or contribute to coinfection with a tumors. CONCLUSION: The review has highlighted the scientific literature focussing on evidence for T. spiralis to act as a pro- or antitumorigenic agent is summarized and discussed, in hope of contributing to a better understanding of the relations between T. spiralis and tumors.

Molecular identification of Trichinella species by multiplex PCR: new insight for Trichinella murrelli.

In order to identify Trichinella at the species level, the commonly used test is a multiplex PCR, allowing the discrimination of nine out of the twelve taxa described so far. This test is based on five primer pairs amplifying fragments of the large subunit rDNA. Each taxon produces one or two bands of different sizes, resulting in a specific band pattern. By multiplex PCR, Trichinella murrelli shows two bands of 127 bp and 316 bp. However, a third band of 256 bp can occur. This band can lead to misidentification, since it is similar to the 253 bp band displayed by Trichinella britovi. BLAST analysis confirmed that the 256 bp band is from T. murrelli. The aim of this short note is to inform analysts that T. murrelli larvae may display either two- or three-band patterns.

TITLE: Identification moléculaire des espèces de Trichinella par PCR multiplex : nouvel éclairage pour Trichinella murrelli. ABSTRACT: Afin d'identifier les Trichinella au niveau de l'espèce, le test couramment utilisé est une PCR multiplex, permettant la discrimination de neuf des douze taxons décrits jusqu'à présent. Ce test est basé sur cinq paires d'amorces amplifiant des fragments de la grande sous-unité l'ADN ribosomal. Chaque taxon produit une ou deux bandes de tailles différentes, résultant en un patron de bandes spécifique. Par PCR multiplex, Trichinella murrelli présente deux bandes de 127 pb et 316 pb. Cependant, une troisième bande de 256 pb peut s'observer. Cette bande peut être la cause d'une erreur d'identification, car elle est similaire à la bande de 253 pb affichée par Trichinella britovi. L'analyse BLAST a confirmé que la bande à 256 pb provient de T. murrelli. Le but de cette note est d'informer les analystes que les larves de T. murrelli peuvent présenter des patrons à deux ou trois bandes.

Immunoproteomic analysis of the excretory-secretory products of Trichinella pseudospiralis adult worms and newborn larvae.

BACKGROUND: The nematode Trichinella pseudospiralis is an intracellular parasite of mammalian skeletal muscle cells and exists in a non-encapsulated form. Previous studies demonstrated that T. pseudospiralis could induce a lower host inflammatory response. Excretory-secretory (ES) proteins as the most important products of host-parasite interaction may play the main functional role in alleviating host inflammation. However, the ES products of T. pseudospiralis early stage are still unknown. The identification of the ES products of the early stage facilitates the understanding of the molecular mechanisms of the immunomodulation and may help finding early diagnostic markers. RESULTS: In this study, we used two-dimensional gel electrophoresis (2-DE)-based western blotting coupled with matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF/TOF-MS/MS) to separate and identify the T. pseudospiralis adult worms ES products immunoreaction-positive proteins. In total, 400 protein spots were separated by 2-DE. Twenty-eight protein spots were successfully identified using the sera from infected pigs and were characterized to correlate with 12 different proteins of T. pseudospiralis, including adult-specific DNase II-10, poly-cysteine and histidine-tailed protein isoform 2, serine protease, serine/threonine-protein kinase ULK3, enolase, putative venom allergen 5, chymotrypsin-like elastase family member 1, uncharacterized protein, peptidase inhibitor 16, death-associated protein 1, deoxyribonuclease II superfamily and golgin-45. Bioinformatic analyses showed that the identified proteins have a wide diversity of molecular functions, especially deoxyribonuclease II (DNase II) activity and serine-type endopeptidase activity. CONCLUSIONS: Early candidate antigens from the ES proteins of T. pseudospiralis have been screened and identified. Our results suggest these proteins may play key roles during the T. pseudospiralis infection and suppress the host immune response. Further, they are the most likely antigen for early diagnosis and the development of a vaccine against the parasite.

Recombinant Trichinella pseudospiralis Serine Protease Inhibitors Alter Macrophage Polarization In Vitro.

During parasite infection, serine protease inhibitors secreted by parasites play important roles in suppressing host defenses. However, the mechanism of immune regulation is unclear. In this study, a serpin gene from Trichinella pseudospiralis, named Tp-Serpin, was cloned and expressed, in order to reveal its role in the regulation of the host immune response in T. pseudospiralis infection. The results showed that Tp-Serpin encodes a 43 kDa protein that was recognized by serum from T. pseudospiralis infected mice at 60 days post-infection (dpi). Tp-Serpin was found to be expressed at all developmental stages of T. pseudospiralis. Inhibitory activity analysis showed that recombinant Tp-Serpin (rTp-Serpin) effectively inhibited the hydrolytic activity of porcine pancreatic elastase (elastase P), human neutrophil elastase (elastase H), and mouse mast cell protease-1, but showed little inhibitory for human neutrophil cathepsin G (cathepsin G). Furthermore, rTp-Serpin induced polarization of macrophages toward the alternatively activated phenotype (M2) alone by activation of the signal transducer and activator of transcription 3 signaling pathway, and inhibited lipopolysaccharide-induced classically activation (M1) in vitro. These data preliminarily demonstrate that Tp-Serpin may play an important role in the immunoregulation of T. pseudospiralis infection by activating the M2-polarized signaling pathway.

Immune responses in mice vaccinated with a DNA vaccine expressing serine protease-like protein from the new-born larval stage of Trichinella spiralis.

Trichinella spiralis is a parasitic helminth that can infect almost all mammals, including humans. Trichinella spiralis infection elicits a typical type 2 immune responses, while suppresses type 1 immune responses, which is in favour of their parasitism. DNA vaccines have been shown to be capable of eliciting balanced CD4+ and CD8+ T cell responses as well as humoral immune responses in small-animal models, which will be advantage to induce protective immune response against helminth infection. In this study, serine protease (Ts-NBLsp) was encoded by a cDNA fragment of new-born T. spiralis larvae, and was inserted after CMV promoter to construct a DNA vaccine [pcDNA3·1(+)-Ts-NBLsp]. Ts-NBLsp expression was demonstrated by immunofluorescence. Sera samples were obtained from vaccinated mice, and they showed strong anti-Ts-NBLsp-specific IgG response. Mice immunized with the pcDNA3·1(+)-Ts-NBLsp DNA vaccine showed a 77·93% reduction in muscle larvae (ML) following challenge with T. spiralis ML. Our results demonstrate that the vaccination with pcDNA3·1(+)-Ts-NBLsp plasmid promoted the balance of type 1 and 2 immune responses and produced a significant protection against T. spiralis infection in mice.

Inadequate labeling of pork sausages prepared in Corsica causing a trichinellosis outbreak in France.

Three cases of human trichinellosis due to Trichinella britovi were reported in 2015 in the Southeast of France resulting from consumption of raw pork sausages (figatelli) prepared in Corsica. Fourteen other people ate figatelli from the same batch but were not infected due to the figatelli being well cooked. This is the first reported human trichinellosis outbreak due to consumption of Corsican sausages prepared from uncontrolled pork. Consumption of raw figatelli is a common tradition in Corsica. As a result, the health recommendation to cook the product well is not always applied. In the present case, the figatelli product label was not sufficiently visible to advise consumers of the risks associated with uncooked pork.