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1.
Acta Trop ; 225: 106172, 2022 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-34627760

RESUMO

The excretory-secretory product (ESP) of Trichinella spiralis (T. spiralis) has been reported to inhibit the growth of various tumor cells, but the mechanism is not yet clear. To explore the effect and mechanism of ESP on liver cancer cells, tumor models were established with H22 cells and then infected with T. spiralis. The results showed that the growth of tumors in mice infected with T. spiralis was significantly inhibited. ESP from adult worms or muscle larvae were then incubated with H22 cells in vitro, and it was found that the ESP could inhibit cell proliferation and promote apoptosis. Subsequently, apoptosis-related proteins in stimulated H22 cells were evaluated, and ESP was found to induce cell apoptosis through the mitochondrial pathway. Additionally, Th-related cytokines were investigated in vivo, and the results showed that the levels of Th1 cytokines were significantly increased in the early stage of T. spiralis infection, while Th2 cytokines increased later than Th1 cytokines, implying that Th1 cytokines with antitumor effects may play a role in inhibiting tumor growth at early stage. In short, ESP can directly induce tumor cell apoptosis and indirectly inhibit tumor cell growth through the host immune system, which may be the antitumor mechanism of T. spiralis infection.


Assuntos
Trichinella spiralis , Triquinelose , Animais , Antígenos de Helmintos , Apoptose , Proliferação de Células , Imunidade , Larva , Camundongos
2.
Parasitol Int ; 86: 102449, 2022 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-34481946

RESUMO

Trichinellosis is a potentially deadly parasitic zoonosis that is contracted by consuming undercooked infected meat. Reliable detection of infectious Trichinella spp. larvae in meat is therefore pivotal to ensure consumer's safety. The recently authorised PrioCHECK™ Trichinella Alternative Artificial Digestion (AAD) test kit appears promising when used with the standard magnetic stirrer method, but evaluation with other apparatus types is lacking. In this study, the performance of the AAD kit in an adapted Trichomatic-35 (TM35) instrument was evaluated, first, at the Swiss National Reference Laboratory for trichinellosis (NRL); second, in a ring trial involving four Swiss official laboratories. Proficiency pork samples spiked with larvae of Trichinella spiralis, T. britovi, or T. pseudospiralis were tested with the AAD kit and with the reference pepsin-HCl digestion method in TM35 instruments. At the NRL, both methods yielded identical qualitative and similar quantitative results independently of the Trichinella species. In the ring trial, satisfactory results were obtained for 47/50 (94.0%) (AAD) and 62/67 (92.5%) (reference method) of the analysed samples. Technical problems impairing analysis were more frequently observed with the AAD kit (n = 22) than with the reference method (n = 5) and were mainly (16/22) reported by one of the external labs. When no technical issues were recorded, the performance of both methods was comparable, in agreement with the observations at the NRL; however, these results suggest a need for further training with the kit and standardisation of the adapted TM35 instruments.


Assuntos
Testes Diagnósticos de Rotina/instrumentação , Parasitologia de Alimentos , Carne de Porco/parasitologia , Trichinella/isolamento & purificação , Animais , Larva/crescimento & desenvolvimento , Sensibilidade e Especificidade , Trichinella/crescimento & desenvolvimento , Trichinella spiralis/crescimento & desenvolvimento , Trichinella spiralis/isolamento & purificação
3.
Rev Bras Parasitol Vet ; 30(4): e012021, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34730610

RESUMO

Trichinellosis is a zoonosis results from eating raw or semi-cooked meat of infected animals. Medicinal plants have been used lately as alternatives and/or combined therapies to resolve some drawbacks of the current regimens. This work analyzed the effect of albendazole monotherapy on Trichinella spiralis experimental infection (group A), in comparison to P. granatum and amygdalin extracts +cobalamin (group B), plus its combination with albendazole (group C). The study revealed that the extracts alone or combined with albendazole had an inferior effect to albendazole monotherapy regarding number of adult worms (40.83 ±3.82, 18.67 ±1.86 and 16.83 ±2.32, respectively). However, their effect was more obvious in muscle phase combined with albendazole, achieving the lower number of larvae/mL tissue homogenate (22.33 ±3.27 in comparison to 39.67 ±2.58 achieved by albendazole monotherapy). The extracts exerted a significant immunomodulatory effect by reducing the local CD4+ expression in the intestine as well as in muscle phase (1.15 ±0.25 and 3.80 ±0.65 in comparison to 4.97 ±0.37 and 12.20 ±0.87 with albendazole monotherapy, respectively). So, these extracts improved the therapeutic efficacy of albendazole, specifically in muscle phase and counteracted the inflammatory reaction caused by albendazole monotherapy, thus extensively alleviating the resulting myositis.


Assuntos
Amigdalina , Miosite , Romã (Fruta) , Trichinella spiralis , Triquinelose , Albendazol , Animais , Modelos Animais de Doenças , Larva , Miosite/veterinária , Extratos Vegetais , Triquinelose/tratamento farmacológico , Triquinelose/veterinária , Vitamina B 12
4.
Ann Parasitol ; 67(2): 195-202, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34592086

RESUMO

Trichinellosis is a common parasitic zoonosis. Complications of anthelmintic drugs combined with steroids raise the urge of alternative protective ways. The study aimed to investigate the protective effects of Lactobacillus acidophilus probiotic on both Trichinella spiralis adults and larvae in experimental animal models. Thirty-six male BALB/c mice were divided into 3 groups: negative control Group (G I); Group (G II) mice were inoculated orally by 500 Trichinella spiralis larvae; tested Group (G III) mice were prophylactic by an oral dose of Lactobacillus acidophilus in commercially available form for seven consecutive days, before infection. Mature worms and encysted larvae were counted on the 5th and 21st day post-infection (dpi), respectively. IL-1, IL-6, IL-10 and TNF-α concentrations were estimated at 5th and 21st dpi of all groups. Significant reductions in mean worms and larvae burden were detected by 62.1% and 73.5% in the prophylactic group compared to the non-prophylactic group. The cytokine profiles were revealed IL-1 and IL-6 up-regulation compared to IL-10 and TNF-α down-regulation in the tested group compared to other groups. Although Lactobacillus acidophilus failed to achieve complete eradication of Trichinella spiralis adults and larvae, it showed powerful effects in reducing parasites and cytokines burdens.


Assuntos
Probióticos , Trichinella spiralis , Triquinelose , Animais , Estudos de Viabilidade , Lactobacillus acidophilus , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Triquinelose/prevenção & controle
5.
Acta Trop ; 224: 106125, 2021 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-34508714

RESUMO

Trichinella spiralis is a major foodborne zoonotic parasitic nematode which has a serious threat to meat food safety. Development of anti-Trichinella vaccine is requisite for control and elimination of Trichinella infection in food animals to ensure meat safety. Aminopeptidase P (TsAPP) and cathepsin X (TsCX) are two novel proteins identified in T. spiralis intestinal infectious L1 larvae (IIL1). The objective of this study was to investigate the protective immunity elicited by immunization with TsAPP and TsCX alone and TsAPP-TsCX in combination in a mouse model. The results demonstrate that subcutaneous vaccination of mice with rTsAPP, rTsCX or rTsAPP + rTsCX elicited a systemic humoral response (high levels of serum IgG, IgG1/IgG2a and IgA) and significant local gut mucosal sIgA responses. The vaccination with rTsAPP, rTsCX or rTsAPP + rTsCX also induced a systemic and local mixed Th1/Th2 response, as demonstrated by clear elevation levels of IFN-γ and IL-4 in vaccinated mice. Vaccination of mice with rTsAPP+rTsCX exhibited a 63.99 % reduction of intestinal adult worms and 68.50% reduction of muscle larva burdens, alleviated inflammation of intestinal mucosal and muscle tissues, and provided a higher immune protection than that of vaccination with rTsAPP or rTsCX alone. The results demonstrated that TsAPP and TsCX might be considered novel candidate target molecules for anti-Trichinella vaccines.


Assuntos
Trichinella spiralis , Triquinelose , Aminopeptidases , Animais , Anticorpos Anti-Helmínticos , Antígenos de Helmintos , Camundongos , Camundongos Endogâmicos BALB C , Triquinelose/prevenção & controle , Vacinação
6.
J Helminthol ; 95: e53, 2021 Sep 13.
Artigo em Inglês | MEDLINE | ID: mdl-34515021

RESUMO

Our work aimed to evaluate the possible effect of Annona muricata (Graviola) leaf extract on Trichinella spiralis in in vitro and in vivo studies. Trichinella spiralis worms were isolated from infected mice and transferred to three culture media - group I (with no drugs), group II (contained Graviola) and group III (contained albendazole) - then they were examined using the electron microscope. In the in vivo study, mice were divided into five groups: GI (infected untreated), GII (prophylactically treated with Graviola for seven days before infection), GIII (infected and treated with Graviola), GIV (infected and treated with albendazole) and GV (infected and treated with a combination of Graviola plus albendazole in half doses). Drug effects were assessed by adults and larvae load beside the histopathological small intestinal and muscular changes. A significant reduction of adult and larval counts occurred in treated groups in comparison to the control group. Histopathologically, marked improvement in the small intestinal and muscular changes was observed in treated groups. Also, massive destruction of the cultured adults' cuticle was detected in both drugs. This study revealed that Graviola leaves have potential activity against trichinellosis, especially in combination with albendazole, and could serve as an adjuvant to anti-trichinellosis drug therapy.


Assuntos
Annona , Trichinella spiralis , Triquinelose , Albendazol/farmacologia , Animais , Modelos Animais de Doenças , Camundongos , Triquinelose/tratamento farmacológico
7.
Vet Parasitol ; 297: 109544, 2021 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-34389193

RESUMO

Processing of meat is one possible approach to control meat-borne parasites. Processing methods such as freezing, cooking and irradiation are recommended for the control of Trichinella in pork, horse or game meat if specific technical conditions are fulfilled. Curing is a widely used preservation process influencing product characteristics such as shelf life, food safety, and taste. As curing methods are characterized by high parameter variability and predictions about inactivation of parasitic stages in raw meat products are difficult, curing and smoking are not recommended for Trichinella control. The objective of this study was to investigate the survival of T. spiralis in cured raw sausages taking into account water activity (aw-value), pH value, temperature, and time. For this purpose, four different types of sausage (Knackwurst, vacuum packed Knackwurst, short ripened salami, long ripened salami) were produced using T. spiralis infested batter. After production, the sausages were stored at product specific conditions for up to 35 days. During storage, pH value and aw-value of the sausages were monitored over time. Further, sausages of each type were digested using the magnetic stirrer method and the viability of the isolated larvae was assessed using a previously published larval motility test as a proxy for viability and infectivity of Trichinella larvae. In this context, we also introduce a three-level rated infectivity score (RIS) with a clear categorization scheme allowing the assessment of the infectivity of larvae. Based on the RIS, larvae isolated from the salamis were regarded as potentially infective until day 2 (short ripened salami) or day 3 (long ripened salami) post ripening, whereas in Knackwurst, potentially infective larvae were still found by day 8 post ripening. In contrast potentially infective larvae were detected in vacuum-packed Knackwurst until day 24 post ripening. Finally, using the RIS approach, data from previously published studies were collected and subjected to a correlation analysis to identify matrix factors linked to short Trichinella inactivation times.


Assuntos
Doenças dos Cavalos , Produtos da Carne , Trichinella spiralis , Trichinella , Triquinelose , Animais , Congelamento , Cavalos , Carne , Triquinelose/veterinária
8.
Vet Parasitol ; 297: 109542, 2021 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-34391612

RESUMO

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.


Assuntos
Doenças dos Roedores , Doenças dos Suínos , Trichinella spiralis , Trichinella , Triquinelose , Animais , Larva , Camundongos , Camundongos Endogâmicos BALB C , Suínos , Triquinelose/veterinária
9.
Vet Parasitol ; 297: 109543, 2021 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-34384643

RESUMO

Trichinellosis is a foodborne parasitic disease caused by Trichinella spp. Different methods, such as cooking, freezing and irradiation, have been suggested to inactivate the parasite in meat infected with Trichinella spp. The International Commission on Trichinellosis (ICT) recommends an irradiation dose of 0.3 kGy to inactivate Trichinella spp. larvae, but its effectiveness in all Trichinella species has not yet been demonstrated. Therefore, the objective of the present study was to determine the effectiveness of gamma radiation in treating wild boar meat to inactivate Trichinella pseudospiralis and Trichinella spiralis. Two wild boars (Sus scrofa) were each inoculated per os (PO) with 20,000 muscle larvae (ML) of Trichinella. One wild boar was inoculated with T. pseudospiralis and the other one with T. spiralis. Both wild boars were euthanized 20 weeks post infection (pi). A 250░g sample from each selected muscle type (side ribs, shoulder and upper forelegs) was obtained in duplicate from each animal. One group of samples was not irradiated (Gc; n░=░6) while the other group of samples was irradiated (Gt; n░=░6) at the Atomic Centre of the Argentinean National Commission of Atomic Energy (CNEA) with a minimum and maximum dose of 0.32-0.41 kGy. Twenty gram of each muscle from Gt and Gc were taken at 24░h, 7 days, 14 days and 21 days post-irradiation, and all samples were individually processed by means of artificial digestion. The ML were then inoculated into mice to evaluate the intestinal phase and the muscular phase. All recovered larvae from Gt and Gc samples showed integrity of the cuticle and active motility. Adult worms and ML were recovered from all mice inoculated with ML from Gc. However, no adult worms or ML were obtained in mice inoculated with ML from Gt. These results show the efficacy of irradiation to inhibit the development of T. pseudospiralis and T. spiralis in the host, which could possibly be extended to other non-encapsulated species, but further studies are needed to demonstrate this hypothesis.


Assuntos
Doenças dos Roedores , Doenças dos Suínos , Trichinella spiralis , Trichinella , Triquinelose , Animais , Raios gama , Carne , Camundongos , Sus scrofa , Suínos , Triquinelose/veterinária
10.
Vet Parasitol ; 298: 109556, 2021 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-34419708

RESUMO

Trichinellosis is a very important food-borne parasitic disease, that seriously endangers animal husbandry and food safety. Therefore, it is necessary to develop a safe and effective vaccine against Trichinella spiralis infection. In this experiment, invasive Lactobacillus plantarum carrying the FnBPA gene served as a live bacterial vector to deliver nucleic acids to the host to produce a novel oral nucleic acid vaccine. Coexpression of the T. spiralis cathepsin F-like protease 1 gene (TsCPF1) and murine IL-4 (mIL-4) by the nucleic acid vaccine was constructed and subsequently delivered to intestinal epithelial cells via invasive L. plantarum. Thirty-seven days after the first immunization, the experimental mice were challenged with 350 T. spiralis infective larvae by oral gavage. The results showed that mice orally immune-stimulated with invasive L. plantarum pValac-TsCPF1/pSIP409-FnBPA not only produce anti-TsCPF1-specific IgG antibodies, sIgA, Th1/Th2 cytokine distinctly increased but also intestinal damage and worm burden relieved compare to non-invasive TsCPF1 group (pValac-TsCPF1/pSIP409). Most notably, experimental mice immunized with invasive L. plantarum coexpressing TsCPF1 and mIL-4 (pValac-TsCPF1-IL-4/pSIP409-FnBPA) exhibited the highest protection efficiency against T. spiralis infection. The above results reveal that invasive L. plantarum-expressing the FnBPA protein improved mucosal and cellular immunity and enhanced resistance to T. spiralis. The nucleic acid vaccine delivered by invasive L. plantarum described in this study offers a novel idea for the prevention of T. spiralis.


Assuntos
Genes de Helmintos , Imunidade , Lactobacillus plantarum , Trichinella spiralis , Triquinelose , Vacinas de DNA , Animais , Genes de Helmintos/genética , Genes de Helmintos/imunologia , Interleucina-4/imunologia , Lactobacillus plantarum/genética , Lactobacillus plantarum/imunologia , Camundongos , Camundongos Endogâmicos BALB C , Trichinella spiralis/imunologia , Triquinelose/prevenção & controle , Triquinelose/veterinária , Vacinas de DNA/administração & dosagem , Vacinas de DNA/imunologia
11.
Mol Immunol ; 138: 87-98, 2021 10.
Artigo em Inglês | MEDLINE | ID: mdl-34364076

RESUMO

Understanding the interaction between the gut microbiota and Trichinella spiralis is of interest for the early diagnosis and development of therapeutics for trichinellosis and to reveal the potential role of microbiota in the mechanism of immunomodulation of this tissue-dwelling helminth. In this study, we utilized 16S rRNA gene sequencing to monitor the dynamics of the microbes in BALB/c mice challenged with T. spiralis. Flow cytometry and ELISA were used to analyze cytokines at the same time. Histopathological analysis of the duodenum was also conducted. We found that microbial perturbations occurred during infection. The abundance of the Lachnospiraceae NK4A136 group, Ruminococcus 1 and Lactococcus decreased. However, the abundance of proinflammatory Parabacteroides increased over time after infection. T. spiralis infection also tended to inhibit IFN-γ production, and promote IL-4 and IL-10 levels. In total, T. spiralis disrupts gut homeostasis and impairs the development of the intestinal ecosystem. Defining the bacterial populations affected by T. spiralis infection might help identify microbial markers for diagnosis of the disease, and the populations could also be further exploited as a novel option to treat T. spiralis infection.


Assuntos
Microbioma Gastrointestinal/imunologia , Imunidade nas Mucosas/imunologia , Triquinelose/imunologia , Triquinelose/microbiologia , Animais , Camundongos , Camundongos Endogâmicos BALB C , Trichinella spiralis/imunologia
12.
PLoS Negl Trop Dis ; 15(8): e0009723, 2021 08.
Artigo em Inglês | MEDLINE | ID: mdl-34432796

RESUMO

OBJECTIVES: Trichinella spiralis is a zoonotic parasite with a complex parasitic life cycle and exposed to animals or humans by infectious meat. To control transmissions of T. spiralis through the food chain to humans, sensitive and selective multihost sera-diagnosis is urgent needed for monitoring T. spiralis exposure. METHODS: A competition enzyme-linked immunosorbent assay (cELISA) for T. spiralis infection diagnosis in multihost sera was developed based on recombinant cystatin-like protein (rCLP-cELISA) as well as monoclonal antibodies. The sensitivity and accuracy of the rCLP-cELISA were quantified using swine (n = 1316), mice (n = 189) and human (n = 157) serum samples. T. spiralis-antibody targeting test ability of the rCLP-cELISA in swine (n = 22) and human (n = 36), instead of other parasites or viruses antibodies, was evaluated. RESULTS: The rCLP-cELISA showed high agreement with commercial ELISA kits in field swine sera assessed by Cohen's kappa value (κ = 0.7963). And it showed 100% specificity in human trichinellosis detection with sensitivity of 96.49%, no cross-reaction with other parasite or virus infections, and high positive detection rate of 87.5% in low-dose infected swine. Besides, the rCLP-cELISA exhibited potential in the detection of T. spiralis, T. nelsoni and Trichinella T8 infections. CONCLUSIONS: The rCLP-cELISA can be used for T. spiralis-associated antibody test in multihost sera.


Assuntos
Anticorpos Anti-Helmínticos/sangue , Cistatinas/imunologia , Ensaio de Imunoadsorção Enzimática/métodos , Doenças dos Suínos/sangue , Trichinella spiralis/isolamento & purificação , Triquinelose/sangue , Animais , Cistatinas/genética , Proteínas de Helminto/genética , Proteínas de Helminto/imunologia , Humanos , Camundongos , Proteínas Recombinantes/genética , Proteínas Recombinantes/imunologia , Suínos , Doenças dos Suínos/parasitologia , Trichinella spiralis/genética , Trichinella spiralis/imunologia , Triquinelose/parasitologia
13.
Acta Trop ; 224: 106112, 2021 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-34453915

RESUMO

Cathepsin L is one member of cysteine protease superfamily and widely distributed in parasitic organisms, it plays the important roles in worm invasion, migration, nutrient intake, molting and immune evasion. The objective of this study was to investigate the biological characteristics of a novel cathepsin L from Trichinella spiralis (TsCL) and its role in larval invasion, development and reproduction. TsCL has a functional domain of C1 peptidase, which belongs to cathepsin L family. The complete TsCL sequence was cloned and expressed in Escherichia coli BL21. The rTsCL has good immunogenicity. RT-PCR and Western blotting analysis showed that TsCL was transcribed and expressed at different T. spiralis phases (e.g., muscle larvae, intestinal infectious larvae, adult worms and newborn larvae). Immunofluorescence test revealed that TsCL was principally localized in the cuticle, stichosome, midgut and female intrauterine embryos of the nematode. rTsCL has the capacity to specially bind with intestinal epithelial cells (IECs) and the binding sites was located in the cytoplasm. rTsCL promoted larval penetration into IEC, while anti-rTsCL antibodies inhibited the invasion. The silencing of TsCL gene by specific dsRNA significantly reduced the TsCL expression and enzyme activity, and also reduced larval invasive ability, development and female reproduction. The results showed that TsCL is an obligatory protease in T. spiralis lifecycle. TsCL participates in worm invasion, development and reproduction, and may be regarded as a potential candidate vaccine/drug target against T. spiralis infection.


Assuntos
Trichinella spiralis , Triquinelose , Animais , Catepsina L , Feminino , Proteínas de Helminto , Larva/genética , Camundongos , Camundongos Endogâmicos BALB C , Reprodução , Trichinella spiralis/genética
14.
Vet Res ; 52(1): 113, 2021 Aug 26.
Artigo em Inglês | MEDLINE | ID: mdl-34446106

RESUMO

The aim of this study was to investigate the biological properties of a novel gut-specific cysteine protease in Trichinella spiralis (TsGSCP) and its role in larval intrusion, development and fecundity. TsGSCP has a functional C1 peptidase domain; C1 peptidase belongs to cathepsin B family. The TsGSCP gene cloned and expressed in Escherichia coli BL21 showed intensive immunogenicity. qPCR and Western blotting revealed that TsGSCP mRNA and protein were expressed at various T. spiralis stages, but their expression levels in intestinal infectious larvae (IIL) were clearly higher than those in muscle larvae (ML), adult worms (AWs) and new-born larvae (NBL). Indirect immunofluorescence (IIF) analysis showed that TsGSCP was primarily located at the outer cuticle and the intrauterine embryos of this parasite. rTsGSCP showed the ability to specifically bind with IECs, and the binding site is within the IEC cytoplasm. rTsGSCP accelerated larval intrusion into host intestinal epithelial cells (IECs), whereas anti-rTsGSCP antibodies suppressed larval intrusion; the acceleration and suppression was induced by rTsGSCP and anti-rTsGSCP antibodies, respectively, in a dose-dependent manner. When ML were transfected with TsGSCP-specific dsRNA, TsGSCP expression and enzymatic activity were reduced by 46.82 and 37.39%, respectively, and the capacity of the larvae to intrude into IECs was also obviously impeded. Intestinal AW burden and adult female length and fecundity were significantly decreased in the group of mice infected with dsRNA-transfected ML compared to the control dsRNA and PBS groups. The results showed that TsGSCP plays a principal role in gut intrusion, worm development and fecundity in the T. spiralis lifecycle and might be a candidate target for vaccine development against Trichinella intrusion and infection.


Assuntos
Cisteína Proteases/genética , Proteínas de Helminto/genética , Trichinella spiralis/fisiologia , Sequência de Aminoácidos , Animais , Cisteína Proteases/química , Cisteína Proteases/metabolismo , Feminino , Fertilidade , Proteínas de Helminto/química , Proteínas de Helminto/metabolismo , Larva/genética , Larva/crescimento & desenvolvimento , Larva/metabolismo , Larva/fisiologia , Camundongos , Filogenia , Alinhamento de Sequência/veterinária , Trichinella spiralis/genética , Trichinella spiralis/crescimento & desenvolvimento , Trichinella spiralis/metabolismo , Triquinelose/veterinária
15.
Acta Trop ; 222: 106071, 2021 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-34331898

RESUMO

A vaccine against Trichinella spiralis infection is urgently needed to interrupt its transmission from domestic animals to humans. However, no vaccine against T. spiralis is currently available. Our previous study demonstrated that the use of the 43-kDa glycoprotein present in excretory-secretory (ES) proteins of muscle larvae (ML) as an intramuscular DNA vaccine led to a 52.1% protection rate against T. spiralis infection. Attenuated Salmonella strains have the advantage of eliciting mucosal immunity, which is important for controlling T. spiralis infections at the intestinal stage and can be provided as vaccines via oral or intranasal routes. Therefore, in this study, complete 43-kDa glycoprotein (Ts43) sequences of T. spiralis were cloned into the vector pYA3681, and the recombinant plasmid pYA3681-Ts43 was transformed into the attenuated Salmonella typhimurium strain χ11802. The results showed that oral vaccination of mice with attenuated Salmonella carrying the recombinant plasmid pYA3681-Ts43 induced an evident elevation of the local intestinal mucosal sIgA and serum IgG antibody responses. The flow cytometry results showed that the percentages of CD4+ T cells and secreted IFN-γ, IL-4, and IL-17A in CD4+ T cells were significantly increased in the spleen and mesenteric lymph node (MLN) lymphocytes of the vaccinated groups. In addition, increased levels of the IFN-γ, IL-4, and IL-17A cytokines were also observed in the serum of the immunized groups. The above immune response results in the immunized groups demonstrated that protective immunity was elicited in this study. Finally, vaccinated mice demonstrated a significant 45.9% reduction in ML burden after infection with T. spiralis. This study demonstrated that oral vaccination with Ts43 delivered by attenuated Salmonella elicited local and systemic concurrent Th1/Th2/Th17 immune responses and provided partial protection against T. spiralis infection in BALB/c mice. This is a prospective strategy for the prevention and control of trichinellosis.


Assuntos
Antígenos de Helmintos , Triquinelose , Vacinas de DNA , Animais , Anticorpos Anti-Helmínticos , Antígenos de Helmintos/genética , Antígenos de Helmintos/imunologia , Camundongos , Camundongos Endogâmicos BALB C , Salmonella typhimurium , Trichinella spiralis/genética , Trichinella spiralis/imunologia , Triquinelose/prevenção & controle , Vacinação
16.
Parasit Vectors ; 14(1): 359, 2021 Jul 09.
Artigo em Inglês | MEDLINE | ID: mdl-34243814

RESUMO

BACKGROUND: Trichinella spiralis ranks seventh in the risk posed by foodborne parasites. It causes most human cases of trichinellosis and is the most frequent cause of Trichinella outbreaks on pig farms and in wild boar, worldwide. Veterinary inspectors seek the source of outbreaks in hopes of limiting the spread. Established molecular tools are inadequate for distinguishing among potential T. spiralis infection sources because genetic variability in these zoonotic pathogens is limited in Europe. Microsatellite markers proved successful in tracing an outbreak of T. britovi, a related parasite harboring much more genetic variation. Here, we successfully employed microsatellite markers to determine the genetic structure of T. spiralis isolates from two pig outbreaks, discovering notable uniformity among parasites within each farm and discovering an epidemiological link between these two outbreaks. METHODS: The individual larvae from five isolates of T. spiralis from two pig farms and from ten wild boars were genotyped using nine microsatellite markers to examine their genetic structure. RESULTS: Notably uniform parasite populations constituted each farm outbreak, and the parasites from the first and second outbreaks resembled each other to a notable degree, indicating an epidemiological link between them. Wild boar harbored more genetically variable larval cohorts, distinguishing them from parasites isolated from domestic pigs. CONCLUSIONS: Microsatellite markers succeeded in distinguishing isolates of the highly homogeneous T. spiralis, aiding efforts to track transmission. Each outbreak was composed of a homogenous group of parasites, suggesting a point source of contamination.


Assuntos
Fazendas/estatística & dados numéricos , Genótipo , Doenças dos Suínos/transmissão , Trichinella spiralis/genética , Triquinelose/transmissão , Triquinelose/veterinária , Animais , Estudos de Coortes , Surtos de Doenças , Repetições de Microssatélites , Polônia/epidemiologia , Sus scrofa/parasitologia , Suínos/parasitologia , Doenças dos Suínos/epidemiologia , Doenças dos Suínos/parasitologia , Trichinella spiralis/classificação , Triquinelose/epidemiologia , Triquinelose/parasitologia
17.
Front Immunol ; 12: 639260, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34093526

RESUMO

The parasitic nematode Trichinella spiralis causes trichinellosis, a serious food-borne parasitic zoonosis worldwide. Infection with T. spiralis may also cause myocarditis. In the present study, we used mouse models to assess the impact of blockage of galectin-receptor interactions by α-lactose on cardiac immunopathology during acute T. spiralis experimental infection. Our data demonstrated that, after T. spiralis infection, blockage of galectin-receptor interactions resulted in cardiac dysfunction detected by transthoracic conventional echocardiography, and increased serum Gal-3 level, a biomarker of myocardial damage. In addition, there were increased eosinophil number in peripheral blood, and increased eosinophil infiltration in the heart and spleen tissues accompanied with increased mRNA levels of eosinophil granule proteins (including eosinophil cationic protein (ECP) and eosinophil peroxidase (EPO)) and IL-5 in these organs; increased cardiac fibrosis accompanied with increased Gal-3 and collagen 1 expressions in the hearts of mice with blockage of galectin-receptor interactions after T. spiralis infection. Correlation analysis showed that significant positive correlations existed between the mRNA levels of Gal-3 and ECP/EPO/eosinophil major basic protein/IL-5/CCL11/CCR3/α-SMA/collagen 1 in the hearts of both T. spiralis-infected mice and T. spiralis-infected mice with blockage of galectin-receptor interactions. Our data suggest that galectin-receptor interactions play a pivotal role during acute T. spiralis infection, and lack of galectin-receptor interactions upregulates Gal-3 which, in turn, leads to elevated heart eosinophil recruitment, exacerbated heart pathology and fibrosis, and heart functional damage.


Assuntos
Galectinas/metabolismo , Cardiopatias/metabolismo , Cardiopatias/patologia , Coração/parasitologia , Trichinella spiralis/parasitologia , Triquinelose/metabolismo , Triquinelose/patologia , Animais , Citocinas/metabolismo , Modelos Animais de Doenças , Eosinofilia/metabolismo , Eosinofilia/parasitologia , Eosinofilia/patologia , Eosinófilos/metabolismo , Eosinófilos/parasitologia , Eosinófilos/patologia , Feminino , Fibrose/metabolismo , Fibrose/parasitologia , Fibrose/patologia , Cardiopatias/parasitologia , Camundongos , RNA Mensageiro/metabolismo , Baço/metabolismo , Baço/parasitologia , Baço/patologia , Triquinelose/parasitologia , Regulação para Cima/fisiologia
18.
Vet Res ; 52(1): 85, 2021 Jun 11.
Artigo em Inglês | MEDLINE | ID: mdl-34116710

RESUMO

Trichinellosis, which is caused by nematodes of the genus Trichinella, is one of the most important zoonotic parasite diseases in the world. A rapid and sensitive immunochromatographic strip (ICS) based on Eu (III) nanoparticles (EuNPs) was developed for the detection of Trichinella spiralis (T. spiralis) infection in pigs. T. spiralis muscle larvae excretory secretory or preadult worm excretory secretory (ML-ES or PAW-ES) antigens were conjugated with EuNPs probes to capture T. spiralis-specific antibodies in pig sera, after which the complex bound to mouse anti-pig IgG deposited on the test line (T-line), producing a fluorescent signal. In the pigs infected with 100, 1000 and 10 000 ML, seroconversion was first detectable for the EuNPs-ML-ES ICS at 30, 25 and 21 days post-infection (dpi) and for the EuNPs-PAW-ES ICS at 25, 21 and 17 dpi. These results show that EuNPs-PAW-ES ICS detects anti-Trichinella IgG in pigs 4-5 days earlier that test using ML-ES antigens. Our ICS have no cross reaction with other parasite infection sera. Furthermore, the detection process could be completed in 10 min. This study indicated that our ICS can be used for the detection of the circulating antibodies in early T. spiralis infection and provide a novel method for on-site detection of T. spiralis infection in pigs.


Assuntos
Anticorpos Anti-Helmínticos/análise , Corantes Fluorescentes/química , Imunoensaio/veterinária , Imunoglobulina G/análise , Doenças dos Suínos/diagnóstico , Trichinella spiralis/isolamento & purificação , Triquinelose/veterinária , Animais , Imunoensaio/métodos , Sus scrofa , Suínos , Doenças dos Suínos/parasitologia , Triquinelose/diagnóstico , Triquinelose/parasitologia
19.
PLoS Negl Trop Dis ; 15(5): e0009408, 2021 05.
Artigo em Inglês | MEDLINE | ID: mdl-33970910

RESUMO

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


Assuntos
Serina Proteases/imunologia , Doenças dos Suínos/imunologia , Trichinella spiralis/enzimologia , Triquinelose/veterinária , Animais , Anticorpos Anti-Helmínticos , Citocinas/sangue , Proteínas de Helminto/genética , Proteínas de Helminto/imunologia , Imunidade Celular , Imunidade Humoral , Músculos/parasitologia , Serina Proteases/genética , Sus scrofa , Suínos , Doenças dos Suínos/prevenção & controle , Trichinella spiralis/genética , Trichinella spiralis/crescimento & desenvolvimento , Triquinelose/imunologia , Vacinação/veterinária
20.
Parasitol Res ; 120(7): 2641-2658, 2021 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-33945012

RESUMO

Ivermectin (IVM) is one of the competitive treatments used for trichinellosis. However, several studies linked its efficacy with early diagnosis and administration to tackle the intestinal phase with limited activity being recorded against encysted larvae. The aim of this study was to employ niosomes for enhancing effectiveness of oral IVM against different stages of Trichinella spiralis (T. spiralis) infection with reference to nano-crystalline IVM. Mice were randomized into four groups: group Ι, 15 uninfected controls; group ΙΙ, 30 infected untreated controls; group ΙΙΙ, 30 infected nano-crystalline IVM treated, and group ΙV, 30 infected niosomal IVM treated. All groups were equally subdivided into 3 subgroups; (a) treated on the 1st day post infection (dpi), (b) treated on the 10th dpi, and (c) treated on the 30th dpi. Assessment was done by counting adult worms and larvae plus histopathological examination of jejunum and diaphragm. Biochemical assessment of oxidant/antioxidant status, angiogenic, and inflammatory biomarkers in intestinal and muscle tissues was also performed. Both niosomes and nano-crystals resulted in significant reduction in adult and larval counts compared to the infected untreated control with superior activity of niosomal IVM. The superiority of niosomes was expressed further by reduction of inflammation in both jejunal and muscle homogenates. Biochemical parameters showed highly significant differences in all treated mice compared to infected untreated control at different stages with highly significant effect of niosomal IVM. In conclusion, niosomal IVM efficacy exceeded the nano-crystalline IVM in treatment of different phases of trichinellosis.


Assuntos
Antiparasitários/administração & dosagem , Ivermectina/administração & dosagem , Trichinella spiralis/efeitos dos fármacos , Triquinelose/tratamento farmacológico , Animais , Antiparasitários/farmacocinética , Antiparasitários/uso terapêutico , Cromatografia Líquida de Alta Pressão , Diafragma , Inflamação/patologia , Ivermectina/farmacologia , Ivermectina/uso terapêutico , Jejuno/patologia , Larva/efeitos dos fármacos , Lipossomos , Masculino , Camundongos , Nanopartículas , Distribuição Aleatória , Trichinella spiralis/fisiologia , Triquinelose/diagnóstico , Zoonoses
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