Trichinella spiralis (Owen, 1835) Induces Increased Dystrophin Expression in Invaded Cross-striated Muscle.

PURPOSE: Dystrophin and the dystrophin glycoprotein complex serve as a cytoskeletal integrator, critical for muscle membrane stability. The aim of the present study was to clarify the expression of dystrophin protein and mRNA in the skeletal muscle tissue during the muscle phase of trichinellosis in mice. METHODS: Muscle tissue was collected from mice experimentally infected with Trichinella spiralis at days 0, 14 and 40 after infection. The expression of dystrophin in the muscle tissue was investigated by immunohistochemistry with antibodies against three different domains of the protein, and the expression levels of Dys mRNA by real-time PCR. RESULTS: The presence of dystrophin protein was increased in the de-differentiating cytoplasm at the early stage of muscle infection and was persisting also in the mature Nurse cell harbouring the parasite. It was accompanied by significantly elevated expression of Dys mRNA at days 14 and 40 after infection. CONCLUSION: Our findings indicate that dystrophin plays a role in regeneration of the muscle and in the Nurse cell formation and stability for security of the parasite survival.

The "starry night" (diffuse microcalcific myopathy) - thousands of muscle microcalcifications after 30 years of Trichinella infection detected by ultrasound.

Trichinellosis, a parasitosis transmitted through consumption of raw or undercooked meat from pigs and game animals, is responsible for a specific myositis. The calcifications of infected myocytes and larva can be detected during many years postin-fection. We present the case of a male patient with a history of severe trichinellosis with disease onset 30 years ago, presenting with generalized muscle microcalcifications detected during musculoskeletal ultrasound evaluation. The ultrasound aspect of the muscles was indeed spectacular; hence, the comparison with a "starry night".

Infection, genetics, and evolution of Trichinella: Historical insights and applications to molecular epidemiology.

Genetic variation in pathogen populations provides the means to answer questions in disease ecology and transmission, illuminating interactions between genetic traits, environmental exposures, and disease. Such studies elucidate the phylogeny, evolution, transmission and pathogenesis of viruses, bacteria and parasites. Here, we review how such studies have fostered understanding of the biology and epidemiology of zoonotic nematode parasites in the genus Trichinella spp., which impose considerable economic and health burdens by infecting wildlife, livestock, and people. To use such data to define ongoing chains of local transmission and source traceback, researchers first must understand the extent and distribution of genetic variation resident in regional parasite populations. Thus, genetic variability illuminates a population's past as well as its present. Here we review how such data have helped define population dynamics of Trichinella spp. in wild and domesticated hosts, creating opportunities to harness genetic variation in the quest to prevent, track, and contain future outbreaks.

Trichinella pseudospiralis infection in red-eared slider, Trachemys scripta, influenced by environmental temperature.

Experimental infection with Trichinella pseudospiralis larvae in red-eared slider Trachemys scripta was carried out. Ten turtles were divided into 3 groups and kept at different temperature (38, 32 and 28 °C). The turtles were fed mice muscles which were infected with T. pseudospiralis larvae (444 - 23,013 larvae/mouse), kept in the aforementioned temperature and necropsied at day 30 after infection. At necropsy, the tongue, forelimbs, hindlimbs and spinal column muscles of turtles were removed separately for pepsin digestion, and the larvae counted. Larvae were found from all 3 turtles kept at 38 °C. Larvae per gram of muscle were highest in the tongue. No larvae were recovered from turtles kept at 32 and 28 °C. The result suggested that T. pseudospiralis is able to infect the red-eared slider in a high temperature environment. We speculated that environmental temperature play an important role in altering the physiological condition of the turtle to facilitate the infection of T. pseudospiralis.

Inactivation of Trichinella muscle larvae at different time-temperature heating profiles simulating home-cooking.

BACKGROUND: Trichinellosis is caused by consumption of raw or undercooked meat containing infective Trichinella muscle larvae (ML). Only few studies on heat-inactivation of Trichinella ML are available in literature and more validated data concerning heat inactivation is needed to improve the risk estimation. OBJECTIVE AND METHODS: The aim of the present study was to evaluate the two in vitro methods "staining" and "morphological examination" as proxies for Trichinella ML heat inactivation in comparison with the mouse bioassay method to get more insight in the relationship between heat, heating time and inactivation of Trichinella ML. The second aim was to evaluate whether these methods could replace the bioassay in the light of ongoing animal use reduction in lifescience research. Tubes containing quantified live Trichinella ML were exposed to heat profiles ranging from 40 to 80 °C. Subsequently, inactivation was evaluated using both methylene blue staining and morphological examination, which was validated by bioassay. Results were used to model Trichinella inactivation. RESULTS: Trichinella muscle larvae exposed to 60 °C or higher for 12-12.5 min were not infective to mice. We found that morphological examination was more consistent with the bioassay than methylene blue staining. Modelled inactivation fitted experimental data consistently. Moreover, this study shows that larval Trichinella morphology may be used in situations where bioassays are not possible or prohibited. CONCLUSIONS: The relationship between heat and inactivation of larvae obtained from this study could be used in Trichinella QMRA models to improve quantification of the risk of Trichinella infection.

Experimental infection of tigerfish (Hydrocynus vittatus) and African sharp tooth catfish (Clarias gariepinus) with Trichinella zimbabwensis.

Trichinella zimbabwensis naturally infects a variety of reptilian and wild mammalian hosts in South Africa. Attempts have been made to experimentally infect piranha fish with T. zimbabwensis and T. papuae without success. Tigerfish (Hydrocynus vittatus) and African sharp tooth catfish (Clarias gariepinus) are accomplished predators cohabiting with Nile crocodiles (Crocodylus niloticus) and Nile monitor lizards (Varanus niloticus) in southern Africa and are natural hosts of T. zimbabwensis. To assess the infectivity of T. zimbabwensis to these two hosts, 24 African sharp tooth catfish (mean live weight 581.75 ± 249.71 g) randomly divided into 5 groups were experimentally infected with 1.0 ± 0.34 T. zimbabwensis larvae per gram (lpg) of fish. Forty-one tigerfish (mean live weight 298.6 ± 99.3 g) were randomly divided for three separate trials. An additional 7 tigerfish were assessed for the presence of natural infection as controls. Results showed no adult worms or larvae of T. zimbabwensis in the gastrointestinal tract and body cavities of catfish sacrificed at day 1, 2 and 7 post-infection (p.i.). Two tigerfish from one experimental group yielded 0.1 lpg and 0.02 lpg of muscle tissue at day 26 p.i. and 28 p.i., respectively. No adult worms or larvae were detected in the fish from the remaining groups sacrificed at day 7, 21, 28, 33 and 35 p.i. and from the control group. Results from this study suggest that tigerfish could sustain T. zimbabwensis under specific yet unknown circumstances.

Acute phase protein pattern and antibody response in pigs experimentally infected with a moderate dose of Trichinella spiralis, T. britovi, and T. pseudospiralis.

The aim of the present study was to evaluate the acute-phase protein (APP) response in three groups of pigs experimentally infected with a moderate infective dose, i.e. 1000 muscle larvae (ML) of Trichinella spiralis, 3000 ML of Trichinella britovi, and 2000 ML of Trichinella pseudospiralis. Over a 62-day period of infection, we examined the serum level and kinetics of the haptoglobin (Hp), C-reactive protein (CRP), serum amyloid A (SAA), and pig major acute-phase protein (pig-MAP). In addition, to better understand the immune response of pigs experimentally infected with three different species of Trichinella, the kinetics of IgG and IgM antibodies against excretory-secretory (ES) antigens of Trichinella ML were also investigated. In order to assess anti-Trichinella IgG dynamics, we used a commercial and an in-house ELISA based on both heterologous (T. spiralis) and homologous (T. spiralis, T. britovi, and T. pseudospiralis) Trichinella species ES antigens. Among the four APPs analyzed, the concentration of CRP and pig-MAP significantly increased only in T. britovi-infected swine when compared with control pigs. This took place as early as 6 days post-infection (dpi). Hp was the only APP whose concentration significantly increased in pigs infected with T. pseudospiralis, this occurring as late as on day 62 pi. Despite the statistical differences found, increases in pig-MAP, CRP, and Hp levels were rather mild and transitory; none of these proteins were found to be elevated in the serum of all experimental groups of pigs at the same time point after infection. Specific IgG antibodies against ES antigens of Trichinella ML were first detected by the commercial and in-house T. spiralis ML ES-antigen ELISAs on days 30, 36 and 36 pi in pigs experimentally infected with T. spiralis, T. britovi, and T. pseudospiralis, respectively. However, seroconversion in pigs experimentally infected with T. britovi was detected slightly earlier (30 dpi) when the ELISA based on homologous rather than heterologous ES antigens was applied. In serum samples from pigs infected with T. spiralis, statistically significant increases in the level of specific IgM antibodies against T. spiralis ML ES antigens were first detected on day 30 pi and after this time, their concentration began to decrease. No changes in the level of anti-Trichinella IgM were observed in T. britovi- or T. pseudospiralis-infected pigs throughout the entire period of the experiment.

Comparison between Trichinella patagoniensis and Trichinella spiralis infection in BALB/c mice.

In Argentina, trichinellosis is an endemic disease acquired mainly through consumption of raw pork infected with nematodes larvae from the Trichinella genus. For years, the only species involved in outbreaks in humans and pig foci in Argentina was Trichinella spiralis. In 2008 the presence of a new Trichinella taxon from a cougar (Puma concolor) was detected and recorded in the province of Rio Negro, Argentina, and the finding was established as a new species in 2012: Trichinella patagoniensis. To the best of our knowledge, there is no information available on the intestinal phase and antibody response in a susceptible host during T. patagoniensis infection. Therefore, our research has been designed to study experimental infection with T. patagoniensis compared to infection with T. spiralis in BALB/c mice. One hundred and twenty eight BALB/c mice were divided into two groups and individuals in each group were infected per os with 500 larvae of T. patagoniensis or 500 larvae of T. spiralis, respectively. After that, they were euthanized on different days. Adult worm recovery from small intestines and artificial digestion of each carcass was performed. Histopathology of small intestines was performed using hematoxylin-eosin staining. Systemic cytokines and antibody kinetics were evaluated. Intestinal adult worm recovery of T. patagoniensis and T. spiralis took place until day 17 and 25, respectively. Systemic IFN-γ, IL-10, and TNF showed significant variations in T. patagoniensis infected mice. Seroconversion was detected in animals as from 15 days post-infection (pi) for both T. patagoniensis and T. spiralis, reaching the highest OD value at 42 days pi. Similar microscopic lesions were observed in the small intestine from mice infected with the same dose of T. spiralis and T. patagoniensis. Our findings contribute new information regarding the intestinal phase and the antibody kinetics of T. patagoniensis in BALB/c mice.

Differences in larval survival and IgG response patterns in long-lasting infections by Trichinella spiralis, Trichinella britovi and Trichinella pseudospiralis in pigs.

BACKGROUND: Domesticated and wild swine play an important role as reservoir hosts of Trichinella spp. and a source of infection for humans. Little is known about the survival of Trichinella larvae in muscles and the duration of anti-Trichinella antibodies in pigs with long-lasting infections. METHODS: Sixty pigs were divided into three groups of 20 animals and infected with 10,000 larvae of Trichinella spiralis, Trichinella britovi or Trichinella pseudospiralis. Four pigs from each group were sacrificed at 2, 6, 12, 18 and 24 months post-infection (p.i.) and the number of larvae per gram (LPG) of muscles was calculated. Serum samples were tested by ELISA and western blot using excretory/secretory (ES) and crude antigens. RESULTS: Trichinella spiralis showed the highest infectivity and immunogenicity in pigs and larvae survived in pig muscles for up to 2 years p.i. In these pigs, the IgG level significantly increased at 30 days p.i. and reached a peak at about 60 days p.i., remaining stable until the end of the experiment. In T. britovi-infected pigs, LPG was about 70 times lower than for T. spiralis at 2 months p.i. and only very few infecting larvae were detected at 6 months p.i., whereas no larvae were detected at 12, 18 and 24 months p.i. At 6 months p.i., degenerated/calcified larvae and cysts were detected in the muscles by trichinoscopy and histology. The IgG pattern showed by T. britovi-infected pigs was similar to that of T. spiralis-infected pigs, although seroconversion occurred some days later. The larval burden of T. pseudospiralis was slightly greater than for T. britovi at 2 months p.i., but no larvae were detected at 6 and 12 months p.i. In T. pseudospiralis-infected pigs, seroconversion occurred slowly, as in T. britovi-infected pigs. The IgG level showed a significant drop at 6 months p.i. and declining to the cut-off value at 12 months p.i. CONCLUSIONS: The longer survival of T. spiralis in pigs in comparison with the other two species highlights its exceptional dissemination potential. These results provide an explanation of the controversial data collected by parasitological and serological tools in the course of epidemiological investigations.

Trichinella spiralis y Trichinella patagoniensis: estudio de la carga globular aplicando el método de Azul Alcian / Trichinella spiralis and Trichinella patagoniensis: study of globular charge applying Alcian Blue method / Trichinella spiralis e Trichinella patagoniensis: estudo da carga globular pelo método de Azul de Alcian

Resumen Los residuos de ácido siálico de las glicoproteínas de superficie son los principales responsables de la carga negativa eritrocitaria. El objetivo de este trabajo fue estudiar alteraciones de carga globular producidas por Trichinella spiralis y Trichinella patagoniensis. Se trabajó con concentrados de larvas musculares de ambas especies y con eritrocitos frescos. Se incubó el sedimento globular con igual volumen de concentrado larval (1 y 2 horas). Los controles fueron incubados con solución salina. Se aplicó el método de Azul Alcian y se determinó el coeficiente experimental de carga aniónica eritrocitaria (CexpCAE). Los resultados mostraron que la carga disminuyó con el aumento del tiempo de tratamiento para ambas especies. Los valores de CexpCAE de las suspensiones incubadas con T. spiralis fueron menores que con T. patagoniensis, indicando que T. spiralis produjo mayor disminución de carga que T. patagoniensis. Se concluye que la desialización producida por ambas especies no es la misma, lo que sugiere que la relación hospedador-parásito que se establecería in vivo sería distinta.

Abstract The sialic acid residues of the surface glycoproteins are mainly responsible for the erythrocyte negative charge. The objective of this work was to study alterations of globular charge produced by Trichinella spiralis and Trichinella patagoniensis. Work was carried out on muscle larvae concentrates of both species and fresh erythrocytes. The treatment was performed by incubating the globular pellet with equal volume of larval concentrate (1 and 2 hours). Controls were incubated with saline solution. The Alcian Blue method was applied and the experimental coefficient of erythrocyte anion charge (expCEAC) was determined. The results showed that the globular charge decreased with increasing treatment time for both species. The expCEAC values of the suspensions incubated with T. spiralis were lower than with T. patagoniensis, indicating that T. spiralis produced a greater decrease in charge than T. patagoniensis. It is concluded that the desialization produced by both species is not the same, suggesting that the host-parasite relationship that would be established in vivo would be different.

Resumo Os resíduos de ácido siálico das glicoproteínas de superfície são os principais responsáveis pela carga negativa dos eritrócitos. O objetivo desse trabalho foi estudar as alterações da carga globular produzidas por Trichinella spiralis e Trichinella patagoniensis. Trabalhamos com concentrados de larvas musculares de ambas as espécies e com eritrócitos frescos. O tratamento foi realizado incubando o sedimento globular com igual volume de concentrado larval (1 e 2 horas), Os controles foram incubados com solução salina. Foi aplicado o método de Azul de Alcian e se determinou o coeficiente experimental de carga aniônica de eritrócitos (CexpCAE). Os resultados mostraram que a carga diminuiu com o aumento do tempo de tratamento para ambas as espécies. Os valores de CexpCAE das suspensões incubadas com T. spiralis foram menores que com T. patagoniensis, indicando que T. spiralis produziu uma diminuição maior na carga que T. patagoniensis. Conclui-se que a dessalinização produzida por ambas as espécies não é a mesma, sugerindo que a relação hospedeiro parasita que seria estabelecida in vivo é diferente.

The Disease Ecology, Epidemiology, Clinical Manifestations, and Management of Trichinellosis Linked to Consumption of Wild Animal Meat.

Historically, human trichinellosis was caused by Trichinella spiralis and transmitted to humans by consumption of undercooked domestic pork. Today, most cases of trichinellosis are caused by other Trichinella species and transmitted by consumption of raw or undercooked wild game meats. Given the increasing global prevalence of wild animal meat-linked trichinellosis, the objectives of this review are: 1) to describe the life cycle and global distribution of Trichinella worms; 2) to describe the changing epidemiology of trichinellosis; 3) to describe the clinical phases of trichinellosis; 4) to recommend the latest diagnostic tests; and 5) to recommend treatment and prevention strategies. Internet search engines were queried with keywords as subject headings to meet the objectives of this review. Although trichinellosis surveillance systems and laws regulating commercial pork production have limited T spiralis-caused trichinellosis in Europe and the United States, trichinellosis due to consumption of raw and undercooked wild boar and feral hog meat continues to occur throughout Southeast Asia. Trichinellosis due to consumption of raw or undercooked meats of other infected game, such as bear, deer, moose, and walrus, continues to occur worldwide. Only adherence to hygienic practices when preparing wild game meats and cooking wild game meats to recommended internal temperatures can prevent transmission of trichinellosis to humans. Wilderness medicine clinicians should be prepared to advise hunters and the public on the risks of game meat-linked trichinellosis and on how to diagnose and treat trichinellosis to prevent fatal complications.

Epidemiology and hypothetical transmission cycles of Trichinella infections in the Greater Kruger National Park of South Africa: an example of host-parasite interactions in an environment with minimal human interactions.

Knowledge on the epidemiology, host range and transmission of Trichinella spp. infections in different ecological zones in southern Africa including areas of wildlife-human interface is limited. The majority of reports on Trichinella infections in sub-Saharan Africa were from wildlife resident in protected areas. Elucidation of the epidemiology of the infections and the prediction of hosts involved in the sylvatic cycles within specific ecological niches is critical. Of recent, there have been reports of Trichinella infections in several wildlife species within the Greater Kruger National Park (GKNP) of South Africa, which has prompted the revision and update of published hypothetical transmission cycles including the hypothetical options based previously on the biology and feeding behaviour of wildlife hosts confined to the GKNP. Using data gathered from surveillance studies and reports spanning the period 1964-2019, confirmed transmission cycles and revised hypothesized transmission cycles of three known Trichinella species (T. zimbabwensis, Trichinella T8 and T. nelsoni) are presented. These were formulated based on the epidemiological factors, feeding habits of hosts and prevalence data gathered from the GKNP. We presume that the formulated sylvatic cycles may be extrapolated to similar national parks and wildlife protected areas in sub-Saharan Africa where the same host and parasite species are known to occur. The anecdotal nature of some of the presented data confirms the need for more intense epidemiological surveillance in national parks and wildlife protected areas in the rest of sub-Saharan Africa to unravel the epidemiology of Trichinella infections in these unique and diverse protected landscapes.

TITLE: Épidémiologie et cycles de transmission hypothétiques des infections à Trichinella dans le Grand Parc National Kruger en Afrique du Sud : un exemple d'interactions hôtes-parasites dans un environnement où les interactions humaines sont minimales. ABSTRACT: Les connaissances sur l'épidémiologie, la gamme d'hôtes et la transmission des infections à Trichinella spp. dans différentes zones écologiques d'Afrique australe, y compris les zones d'interface faune-homme, sont limitées. La majorité des signalements d'infections à Trichinella en Afrique subsaharienne proviennent animaux sauvages résidant dans des zones protégées. L'élucidation de l'épidémiologie des infections et la prévision des hôtes impliqués dans les cycles sylvatiques au sein de niches écologiques spécifiques sont essentielles. Récemment, des infections à Trichinella ont été signalées chez plusieurs espèces sauvages dans le Grand Parc National Kruger (GPNK) en Afrique du Sud, ce qui a incité à la révision et à la mise à jour des cycles de transmission hypothétiques publiés, y compris les options hypothétiques basées précédemment sur la biologie et le comportement alimentaire des hôtes de la faune sauvage confinés au GPNK. À partir des données des études de surveillance et de rapports couvrant la période 1964­2019, les cycles de transmission confirmés et les cycles de transmission hypothétiques révisés de trois espèces connues de Trichinella (T. zimbabwensis, Trichinella T8 et T. nelsoni) sont présentés. Ceux-ci ont été formulés sur la base des facteurs épidémiologiques, des habitudes alimentaires des hôtes et des données de prévalence recueillies au GPNK. Nous supposons que les cycles sylvatiques formulés peuvent être extrapolés aux parcs nationaux et zones protégées similaires en Afrique subsaharienne, où les mêmes hôtes et espèces de parasites sont connues. La nature anecdotique de certaines des données présentées confirme la nécessité d'une surveillance épidémiologique plus intense dans les parcs nationaux et zones protégées du reste de l'Afrique subsaharienne pour démêler l'épidémiologie des infections à Trichinella dans ces paysages protégés uniques et diversifiés.

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.

Human trichinellosis caused by Trichinella britovi in Greece, and literature review.

Trichinellosis is a cosmopolitan zoonotic parasitic disease caused by the nematodes of the genus Trichinella, through the consumption of raw or semi-raw infected meat from swine, horses and wild animals. This disease has been sporadically reported in Greece since 1946. The aim of the present study was to describe a trichinellosis case in a patient hospitalized in northern Greece, in 2017. A 47-year-old male was admitted to hospital with intense generalized myalgia, periorbital swelling, fever, exhaustion and anorexia. Biochemical and haematological profile showed eosinophilia and elevated creatine phosphokinase (CPK). Anti-Trichinella spp. IgG and IgM antibodies were detected by serology and Trichinella spp. larvae were found in two muscle biopsies by compressorium and histological examination. A larva collected from the muscle biopsy was identified as Trichinella britovi by polymerase chain reaction (PCR). Albendazole (400 mg twice per day × 10 days) was administered and the clinical condition of the patient promptly improved. This is the first identification of T. britovi in a patient in Greece.

Trichinella britovi muscle larvae and adult worms: stage-specific and common antigens detected by two-dimensional gel electrophoresis-based immunoblotting.

BACKGROUND: Trichinella britovi is the second most common species of Trichinella that may affect human health. As an early diagnosis of trichinellosis is crucial for effective treatment, it is important to identify sensitive, specific and common antigens of adult T. britovi worms and muscle larvae. The present study was undertaken to uncover the stage-specific and common proteins of T. britovi that may be used in specific diagnostics. METHODS: Somatic extracts obtained from two developmental stages, muscle larvae (ML) and adult worms (Ad), were separated using two-dimensional gel electrophoresis (2-DE) coupled with immunoblot analysis. The positively-visualized protein spots specific for each stage were identified through liquid chromatography-tandem mass spectrometry (LC-LC/MS). RESULTS: A total of 272 spots were detected in the proteome of T. britovi adult worms (Ad) and 261 in the muscle larvae (ML). The somatic extracts from Ad and ML were specifically recognized by T. britovi-infected swine sera at 10 days post infection (dpi) and 60 dpi, with a total of 70 prominent protein spots. According to immunoblotting patterns and LC-MS/MS results, the immunogenic spots recognized by different pig T. britovi-infected sera were divided into three groups for the two developmental stages: adult stage-specific proteins, muscle larvae stage-specific proteins, and proteins common to both stages. Forty-five Ad proteins (29 Ad-specific and 16 common) and thirteen ML proteins (nine ML-specific and four common) cross-reacted with sera at 10 dpi. Many of the proteins identified in Ad (myosin-4, myosin light chain kinase, paramyosin, intermediate filament protein B, actin-depolymerizing factor 1 and calreticulin) are involved in structural and motor activity. Among the most abundant proteins identified in ML were 14-3-3 protein zeta, actin-5C, ATP synthase subunit d, deoxyribonuclease-2-alpha, poly-cysteine and histide-tailed protein, enolase, V-type proton ATPase catalytic and serine protease 30. Heat-shock protein, intermediate filament protein ifa-1 and intermediate filament protein B were identified in both proteomes. CONCLUSIONS: To our knowledge, this study represents the first immunoproteomic identification of the antigenic proteins of adult worms and muscle larvae of T. britovi. Our results provide a valuable basis for the development of diagnostic methods. The identification of common components for the two developmental stages of T. britovi may be useful in the preparation of parasitic antigens in recombinant forms for diagnostic use.

Ultrastructural preservation of tissues and their reaction to the infection with trichinella in the El Plomo mummy: Muscle fiber ultrastructure and trichinosis/mummy of the Cerro El Plomo.

The El Plomo mummy was a pre-Columbian Incan child who was found mummified in the Andes Mountains above an altitude of 17,700 feet. In the environment, natural mummification occurred due to low temperatures and strong winds. Dating measurements (relative dating) by experts from the National Museum of Natural History of Chile established that the mummified body corresponds the Inca period (1,450 to 1,500 AD). In 2003, the body was transferred to the University of Chile Medical School for exhaustive medical examination. Tissue samples from the right quadriceps muscle were extracted and fixed in glutaraldehyde and postfixed in osmium tetroxide to obtain ultrathin sections to be observed by transmission electron microscope. Images were recorded on photographic paper, digitalized and analyzed by experts on morphology. Results showed a preservation of cell boundaries in striated muscle cells, but specific subcellular organelles or contractile sarcomeric units (actin and myosin) were unable to be recognized. However, the classical ultrastructural morphology of the polypeptide collagen type I was preserved intact both in primary and secondary organization. Therefore, we concluded that the process of natural mummification by freezing and strong winds is capable of damaging the ultrastructure of muscle cells and preserving collagen type I intact.

Parasite to patient: A quantitative risk model for Trichinella spp. in pork and wild boar meat.

Consumption of raw or inadequately cooked pork meat may result in trichinellosis, a human disease due to nematodes of the genus Trichinella. In many countries worldwide, individual control of pig carcasses at meat inspection is mandatory but incurs high costs in relation to absence of positive carcasses from pigs reared under controlled housing. EU regulation 2015/1375 implements an alternative risk-based approach, in view of absence of positive findings in pigs under controlled housing conditions. Moreover, Codex Alimentarius guidelines for the control of Trichinella spp. in meat of suidae have been published (CAC, 2015) and used in conjunction with the OIE terrestrial Animal health code, to provide guidance to governments and industry on risk based control measures to prevent human exposure to Trichinella spp. and to facilitate international pork trade. To further support such a risk-based approach, we model the risk of human trichinellosis due to consumption of meat from infected pigs, raised under non-controlled housing and wild boar, using Quantitative Microbial Risk Assessment (QMRA) methods. Our model quantifies the distribution of Trichinella muscle larve (ML) in swine, test sensitivity at carcass control, partitioning of edible pork parts, Trichinella ML distribution in edible muscle types, heat inactivation by cooking and portion sizes. The resulting exposure estimate is combined with a dose response model for Trichinella species to estimate the incidence of human illness after consumption of infected meat. Paramater estimation is based on experimental and observational datasets. In Poland, which served as example, we estimated an average incidence of 0.90 (95%CI: 0.00-3.68) trichinellosis cases per million persons per year (Mpy) due to consumption of pork from pigs that were reared under non-controlled housing, and 1.97 (95%CI: 0.82-4.00) cases per Mpy due to consumption of wild boar. The total estimated incidence of human trichinellosis attributed to pigs from non-controlled housing and wild boar in Poland, is similar to the incidence of human trichinellosis in that country reported by EFSA. Overall, in Europe, we estimated an upper incidence limit of 5.3×10-4 cases per Mpy, or less than one predicted case of trichinellosis in the European Union every 4years, due to consumption of pork from controlled housing. Therefore, Trichinella testing of pigs under controlled housing is not adding any value to protect human health. We suggest applying our farm-to-fork QMRA model to further support decision making on the global scale.