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We conducted a recent investigation in Quebec, Canada, concerning Canadian deer hunters who went to the United States to hunt deer and returned with symptoms of fever, severe headache, myalgia, and articular pain of undetermined etiology. Further investigation identified that a group of 10 hunters from Quebec attended a hunting retreat in Illinois (USA) during November 22-December 4, 2018. Six of the 10 hunters had similar symptoms and illness onset dates. Serologic tests indicated a recent toxoplasmosis infection for all symptomatic hunters, and the risk factor identified was consumption of undercooked deer meat. Among asymptomatic hunters, 2 were already immune to toxoplasmosis, 1 was not immune, and the immune status of 1 remains unknown. Outbreaks of acute toxoplasmosis infection are rare in North America, but physicians should be aware that such outbreaks could become more common.
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Carne , Toxoplasmose/epidemiologia , Adulto , Animais , Culinária/normas , Cervos , Humanos , Masculino , Carne/parasitologia , Pessoa de Meia-Idade , Quebeque/epidemiologia , Fatores de Risco , Toxoplasmose/sangue , Toxoplasmose/etiologiaRESUMO
The primary aim of this work was to isolate common bovine digestive tract parasites in recycled manure bedding (RMS), as well as to determine the ability of current RMS preparation procedures to eliminate these pathogens. Other objectives were to assess whether any of the aforementioned parasites could be retrieved in bulk milk from dairies using RMS and to study whether the prevalence of these parasites differed among manure of cows housed on RMS versus on straw bedding. For the study, 27 RMS farms and 61 control farms were recruited. Samples of manure from the pre-pit and milk from the bulk tank were recovered from straw-bedding farms and RMS-based farms. In addition, samples from the manure solid fraction after liquid extraction, RMS before use, and RMS currently in use were recovered from RMS herds. Parasites were first detected by double centrifugation zinc sulfate flotation to enhance isolation of gastrointestinal protozoa, and by modified Wisconsin sugar flotation for the appraisal of gastrointestinal nematodes. Cryptosporidium parasites were confirmed by nested PCR amplification and sequencing of a portion of the gene encoding the small subunit rRNA. Results revealed a high prevalence of Cryptosporidium spp. (C. parvum, C. andersoni, and C. meleagridis, identified by PCR) and Eimeria spp. (mainly E. bovis and E. zuernii) parasites in both types of farms, with a larger proportion of manure samples from RMS-bedded farms testing positive for Cryptosporidium parasites compared with manure from straw-bedded farms. Both Cryptosporidium spp. and Eimeria spp. oocysts were found at every step of RMS preparation and transformation, showing that current RMS preparation strategies do not guarantee the destruction of protozoan parasites. Cryptosporidium parvum, a potential zoonotic risk for professionals in close contact with livestock, was found to be present in 32 out of 61 straw-bedded and 24 of 27 RMS farms. No protozoan parasites were found in any sample derived from bulk milk, neither by microscopy analysis nor by molecular methods.
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Doenças dos Bovinos/parasitologia , Cryptosporidium parvum/isolamento & purificação , Indústria de Laticínios/métodos , Eimeria/isolamento & purificação , Enteropatias Parasitárias/veterinária , Esterco , Criação de Animais Domésticos , Animais , Bovinos , Feminino , Enteropatias Parasitárias/parasitologia , Leite/química , Oocistos , ReciclagemRESUMO
BACKGROUND: Lyme disease (LD), a multisystem infection caused by the spirochete Borrelia burgdorferi sensu stricto (B. burgdorferi), is the most reported vector-borne disease in North America, and by 2020, 80% of the population in central and eastern Canada could live in LD risk areas. Among the key factors for minimising the impact of LD are the accurate diagnosis and appropriate management of patients bitten by ticks. In this study, the practices of Quebec general practitioners (GPs) on LD diagnosis and management of patients bitten by infected ticks are described. METHODS: Eight years (2008 to 2015) of retrospective demographic and clinical data on patients bitten by infected Ixodes scapularis (I. scapularis) ticks and on the management of suspected and confirmed LD cases by Quebec GPs were analysed. RESULTS: Among 50 patients, all the antimicrobial treatments of LD clinical cases were appropriate according to current guidelines. However, more than half (62.8%) of erythema migrans (EM) were possibly misdiagnosed, 55.6%, (n = 27) of requested serologic tests were possibly unnecessary and the majority (96.5%, n = 57) of prophylactic antimicrobial treatments were not justified according to current guidelines. CONCLUSIONS: These observations underline the importance for public health to enhance the knowledge of GPs where LD is emerging, to minimise the impact of the disease on patients and the financial burden on the health system.
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Clínicos Gerais/estatística & dados numéricos , Doença de Lyme/diagnóstico , Padrões de Prática Médica/estatística & dados numéricos , Animais , Anti-Infecciosos/uso terapêutico , Humanos , Ixodes , Doença de Lyme/tratamento farmacológico , Doença de Lyme/terapia , Quebeque , Estudos Retrospectivos , Inquéritos e Questionários , Picadas de Carrapatos/complicações , Picadas de Carrapatos/tratamento farmacológico , Picadas de Carrapatos/terapiaRESUMO
Context: Environmental changes will foster the spread of Ixodes scapularis ticks and increase the incidence of Lyme disease in Québec in the coming years. The objective of this study is to estimate the epidemiological and clinical burden and part of the current economic burden of Lyme disease in Québec and to estimate the number of cases expected by 2050. Methods: Cases of Lyme disease reported in Québec from 2015 to 2019 were used to describe their demographic, geographical and clinical characteristics and the cost of their initial care. Three incidence rate scenarios were then developed to estimate the number of cases expected by 2050, based on demographic and climate projections. Results: From 2016 to 2019, 1,473 cases of Lyme disease were reported in Québec. Over 90% of those cases were acquired in two regions of southern Québec (Estrie and Montérégie), while the individuals infected were residents from all over Québec. The average age of cases is 44 years and 66% of infections were at the localized stage, the first stage of Lyme disease. The cost of initial care is estimated at an average of $182 CAN per patient ($47 CAN at the localized stage and $443 CAN at the disseminated stage). According to projections, over 95% of the Québec population will live in a climate zone conducive to the establishment of ticks by 2050, with a number of cases acquired in Québec being 1.3 to 14.5 times higher than in 2019, depending on the incidence rate scenario used. Conclusion: The epidemiological burden is concentrated primarily in southern Québec, but the clinical and economic burden is already distributed throughout the province. The projections for 2050 should help the regions of Québec adapt and optimize public health protection measures.
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Malaria remains a significant risk in many areas of the world, with resistance to the current antimalarial pharmacopeia an ever-increasing problem. The M1 alanine aminopeptidase (PfM1AAP) and M17 leucine aminopeptidase (PfM17LAP) are believed to play a role in the terminal stages of digestion of host hemoglobin and thereby generate a pool of free amino acids that are essential for parasite growth and development. Here, we show that an orally bioavailable aminopeptidase inhibitor, CHR-2863, is efficacious against murine malaria.
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Aminopeptidases/antagonistas & inibidores , Antimaláricos/farmacologia , Inibidores Enzimáticos/farmacologia , Animais , Antimaláricos/química , Inibidores Enzimáticos/química , Feminino , Malária/parasitologia , Camundongos , Camundongos Endogâmicos C57BL , Plasmodium falciparum/efeitos dos fármacos , Plasmodium falciparum/patogenicidadeRESUMO
The incidence of Lyme disease is increasing in Québec and is closely linked to the distribution of Ixodes scapularis ticks. A time-to-establishment model developed in 2012 by Leighton and colleagues predicted the year of tick population establishment for each municipality in eastern Canada. To validate if this model correctly predicted tick distribution in Québec, predicted tick establishment was compared to field data from active tick surveillance (2010-2018) using two criteria: i) the detection of at least one tick and ii) the detection of the three questing stages of the tick. The speed of tick establishment and the increase in the exposed human population by 2100 were predicted with the time-to-establishment model. Field observations were consistent with model predictions. Ticks were detected on average 3 years after the predicted year. The probability of tick detection is significantly higher after the predicted year than before (61% vs 27% of collections). The trend was similar for the detection of three tick stages (16% vs 9% of collections). The average speed of tick range expansion was estimated by the model to be 18 km/year in Québec, with 90% of the human population exposed by 2027. The validation of the time-to-establishment model using field data confirmed that it could be used to project I. scapularis range expansion in Québec, and consequently the increase in Lyme disease risk over the coming decades. This will help public health authorities anticipate and adapt preventive measures, especially in areas not yet affected by Lyme disease.
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Ixodes/microbiologia , Ixodes/fisiologia , Migração Animal , Animais , Vetores Aracnídeos , Borrelia burgdorferi , Sistemas de Informação Geográfica , Geografia , Humanos , Doença de Lyme/epidemiologia , Saúde Pública , Quebeque , Reprodutibilidade dos Testes , Infestações por Carrapato/epidemiologiaRESUMO
BACKGROUND: Despite increases in cases of Lyme disease, little is known about the management and clinical course of the disease in Canada. We aimed to describe the management and clinical course of Lyme disease in patients treated in acute care facilities in Quebec and to assess adherence to the 2006 Infectious Diseases Society of America (IDSA) guideline. METHODS: This retrospective multicentre cohort study included pediatric and adult patients with serologically confirmed Lyme disease treated in acute care facilities (12 community hospitals and 2 tertiary care centres) of 2 endemic regions of Quebec (Estrie and Montérégie), from 2004 to 2017. We considered drug choice, prescribed dose and treatment duration in assessing adherence of prescriptions to the 2006 IDSA guideline. The main outcome was complete resolution of symptoms at 3 months after the initiation of treatment. RESULTS: We included 272 patients from 14 institutions (age range 3-87 yr). Early disseminated Lyme disease (140 patients [51%]) was predominant. Adherence to the IDSA guideline was observed in 235 (90%) of the 261 cases with complete information, and adherence was stable over time (2004-2013: 57/64 [89%]; 2014-2015: 64/71 [90%]; 2016-2017: 114/126 [90%]; p = 0.8). Non-adherence to the guideline (n = 26) was predominantly due to longer-than-recommended treatment duration (16/26 [62%]). Resolution of objective signs at 3 months after treatment initiation occurred in 265 (99%) of 267 patients, whereas post-treatment Lyme disease syndrome was observed in 27 patients (10%) with increasing incidence over time (2004-2013: 3/65 [5%]; 2014-2015: 4/73 [5%]; 2016-2017: 20/129 [16%]; p = 0.02). INTERPRETATION: We observed clinical resolution of Lyme disease in 99% of the patients, and most treatments (90%) complied with the 2006 IDSA guideline. The incidence of post-treatment Lyme disease syndrome increased over the study period, warranting further prospective studies.
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Doença de Lyme , Síndrome Pós-Lyme , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Canadá , Criança , Pré-Escolar , Estudos de Coortes , Humanos , Doença de Lyme/diagnóstico , Doença de Lyme/tratamento farmacológico , Doença de Lyme/epidemiologia , Pessoa de Meia-Idade , Estudos Prospectivos , Quebeque/epidemiologia , Estudos Retrospectivos , Adulto JovemRESUMO
Cyclospora cayetanensis is an emerging foodborne parasite that causes cyclosporiasis, an enteric disease of humans. Domestically acquired outbreaks have been reported in Canada every spring or summer since 2013. To date, investigations into the potential sources of infection have relied solely on epidemiological data. To supplement the epidemiological data with genetic information, we genotyped 169 Canadian cyclosporiasis cases from stool specimens collected from 2010 to 2021 using an existing eight-marker targeted amplicon deep (TADS) scheme specific to C. cayetanensis as previously described by the US Centers for Disease Control and Prevention (CDC). This is the first study to genotype Canadian Cyclospora cayetanensis isolates, and it focuses on evaluating the genotyping performance and genetic clustering. Genotyping information was successfully collected with at least part of one of the markers in the TADS assay for 97.9% of specimens, and 81.1% of cyclosporiasis cases met the minimum requirements to genetically cluster into 20 groups. The performance of the scheme suggests that examining cyclosporiasis cases genetically will be a valuable tool for supplementing epidemiological outbreak investigations and to minimize further infections. Further research is required to expand the number of discriminatory markers to improve genetic clustering.
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Lyme disease (LD) is an emerging public health threat in Canada, associated with the northward range expansion of the black-legged tick (Ixodes scapularis). To address this, public health authorities have been carrying out surveillance activities and awareness campaigns targeting vulnerable populations such as outdoor workers. Implementing these measures is time-consuming and resource-intensive, prompting the assessment of alternatives. Our goal was to evaluate the feasibility and implementation of a training-of-trainers-inspired approach in raising awareness about LD risk and prevention among workers and general population, as well as to evaluate its potential to contribute to provincial LD surveillance efforts. We trained a group of workers from publicly-accessible outdoor parks of the province of Québec to become "LD education ambassadors". Ambassadors were trained to raise tick and LD awareness, share information on preventive measures in their respective communities, and lead tick sampling activities using a standardised protocol similar to that used by Public Health authorities. Ambassador-led outreach activities, public reach, sampling activities and collected ticks were documented, as well as ambassadors' satisfaction with the training using forms and semi-structured interviews. In total, 18 ambassadors from 12 organizations were trained. Between June and September 2019, they led 28 independent outreach activities, reaching over 1 860 individuals (from occupational and general public settings) in seven public health units. Ambassadors led 28 tick samplings, together collecting 11 I. scapularis ticks. This study suggests that an adapted training-of-trainers is a feasible approach to raising tick and LD risk awareness among Québec outdoor workers and public. Trained ambassadors have the potential of reaching a large portion of the population visiting or working in outdoor parks while also providing much-needed outreach regarding risk and prevention. Pushing this concept further to include other types of workers and jurisdictions may contribute to national LD surveillance efforts.
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Doença de Lyme/prevenção & controle , Voluntários/educação , Adulto , Animais , Canadá , Feminino , Humanos , Ixodes/fisiologia , Doença de Lyme/parasitologia , Masculino , Pessoa de Meia-Idade , Vigilância da População/métodos , QuebequeRESUMO
Cats that spend time outdoors and dogs are particularly at risk of exposure to ticks and the pathogens they transmit. A retrospective study on data collected through passive tick surveillance was conducted to estimate the risk of exposure to tick-borne pathogens in cats and dogs bitten by blacklegged ticks (Ixodes scapularis) in the province of Quebec, Canada, from 2010 to 2017. Blacklegged ticks collected from these host animals were tested by PCR for Borrelia burgdorferi sensu stricto, Borrelia miyamotoi, Anaplasma phagocytophilum, and Babesia microti. A total of 13,733 blacklegged ticks were collected from 12,547 animals. Most ticks were adult females and partially engorged. In total, 1,774 cats were infested with ticks and 22.6 and 2.7% of these animals were bitten by at least one tick infected with B. burgdorferi and A. phagocytophilum, respectively. For the 10,773 tick infested dogs, 18.4% were exposed to B. burgdorferi positive ticks while 1.9% of infested dogs were exposed to ticks infected with A. phagocytophilum. The risk of exposure of both cats and dogs to B. miyamotoi and B. microti was lower since only 1.2 and 0.1% of ticks removed were infected with these pathogens, respectively. Traveling outside of the province of Quebec prior to tick collection was significantly associated with exposure to at least one positive tick for B. burgdorferi, A. phagocytophilum and B. microti. Animals exposed to B. burgdorferi or B. miyamotoi positive tick(s) were at higher risk of being concurrently exposed to A. phagocytophilum; higher risk of exposure to B. microti was also observed in animals concurrently exposed to B. burgdorferi. The odds of dogs having B. burgdorferi antibodies were higher when multiple ticks were collected on an animal. The testing and treatment strategies used on dogs bitten by infected ticks were diverse, and misconceptions among veterinarians regarding the treatment of asymptomatic but B. burgdorferi-seropositive dogs were noted. In conclusion, our study demonstrates that cats and dogs throughout Quebec are exposed to blacklegged ticks infected with B. burgdorferi and A. phagocytophilum, and veterinarians across the province need to be aware of this potential threat to the health of pets and their owners.
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The Plasmodium falciparum M1 alanyl aminopeptidase and M17 leucyl aminopeptidase, PfM1AAP and PfM17LAP, are potential targets for novel anti-malarial drug development. Inhibitors of these aminopeptidases have been shown to kill malaria parasites in culture and reduce parasite growth in murine models. The two enzymes may function in the terminal stages of haemoglobin digestion, providing free amino acids for protein synthesis by the rapidly growing intra-erythrocytic parasites. Here we have performed a comparative cellular and biochemical characterisation of the two enzymes. Cell fractionation and immunolocalisation studies reveal that both enzymes are associated with the soluble cytosolic fraction of the parasite, with no evidence that they are present within other compartments, such as the digestive vacuole (DV). Enzyme kinetic studies show that the optimal pH of both enzymes is in the neutral range (pH 7.0-8.0), although PfM1AAP also possesses some activity (< 20%) at the lower pH range of 5.0-5.5. The data supports the proposal that PfM1AAP and PfM17LAP function in the cytoplasm of the parasite, likely in the degradation of haemoglobin-derived peptides generated in the DV and transported to the cytosol.
Assuntos
Antígenos CD13/metabolismo , Leucil Aminopeptidase/metabolismo , Plasmodium falciparum/enzimologia , Proteínas de Protozoários/metabolismo , Animais , Antimaláricos/farmacologia , Antimaláricos/uso terapêutico , Antígenos CD13/antagonistas & inibidores , Antígenos CD13/química , Antígenos CD13/isolamento & purificação , Fracionamento Celular , Células Cultivadas , Citosol/enzimologia , Desenvolvimento de Medicamentos , Ensaios Enzimáticos , Eritrócitos/parasitologia , Humanos , Concentração de Íons de Hidrogênio , Leucil Aminopeptidase/antagonistas & inibidores , Leucil Aminopeptidase/química , Leucil Aminopeptidase/isolamento & purificação , Malária Falciparum/tratamento farmacológico , Malária Falciparum/parasitologia , Plasmodium falciparum/efeitos dos fármacos , Proteínas de Protozoários/antagonistas & inibidores , Proteínas de Protozoários/química , Proteínas de Protozoários/isolamento & purificação , Coelhos , Proteínas Recombinantes/química , Proteínas Recombinantes/isolamento & purificaçãoRESUMO
BACKGROUND: Cryptosporidiosis is a gastrointestinal disease with global distribution. It has been a reportable disease in Canada since 2000; however, routine molecular surveillance is not conducted. Therefore, sources of contamination are unknown. The aim of this project was to identify species and subtypes of Cryptosporidium in clinical cases from Ontario, the largest province in Canada, representing one third of the Canadian population, in order to understand transmission patterns. METHODS: A total of 169 frozen, banked, unpreserved stool specimens that were microscopy positive for Cryptosporidium over the period 2008-2017 were characterized using molecular tools. A subset of the 169 specimens were replicate samples from individual cases. DNA was extracted directly from the stool and nested PCR followed by Sanger sequencing was conducted targeting the small subunit ribosomal RNA (SSU) and glycoprotein 60 (gp60) genes. RESULTS: Molecular typing data and limited demographic data were obtained for 129 cases of cryptosporidiosis. Of these cases, 91 (70.5 %) were due to Cryptosporidium parvum and 24 (18.6%) were due to Cryptosporidium hominis. Mixed infections of C. parvum and C. hominis occurred in four (3.1%) cases. Five other species observed were Cryptosporidium ubiquitum (n = 5), Cryptosporidium felis (n = 2), Cryptosporidium meleagridis (n = 1), Cryptosporidium cuniculus (n = 1) and Cryptosporidium muris (n = 1). Subtyping the gp60 gene revealed 5 allelic families and 17 subtypes of C. hominis and 3 allelic families and 17 subtypes of C. parvum. The most frequent subtype of C. hominis was IbA10G2 (22.3%) and of C. parvum was IIaA15G2R1 (62.4%). CONCLUSIONS: The majority of isolates in this study were C. parvum, supporting the notion that zoonotic transmission is the main route of cryptosporidiosis transmission in Ontario. Nonetheless, the observation of C. hominis in about a quarter of cases suggests that anthroponotic transmission is also an important contributor to cryptosporidiosis pathogenesis in Ontario.
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Cryptosporidium/classificação , Cryptosporidium/genética , Fezes/parasitologia , Variação Genética , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Criança , Pré-Escolar , Criptosporidiose/epidemiologia , Criptosporidiose/parasitologia , Criptosporidiose/transmissão , Cryptosporidium/isolamento & purificação , DNA de Protozoário/genética , Feminino , Genótipo , Humanos , Masculino , Pessoa de Meia-Idade , Ontário/epidemiologia , Filogenia , Análise de Sequência de DNA , Adulto JovemRESUMO
INTRODUCTION: Climate change plays an important role in the geographic spread of zoonotic diseases. Knowing which populations are at risk of contracting these diseases is critical to informing public health policies and practices. In Québec, 14 zoonoses have been identified as important for public health to guide the climate change adaptation efforts of decision-makers and researchers. A great deal has been learned about these diseases in recent years, but information on at-risk workplaces remains incomplete. The objective of this study is to paint a portrait of the occupations and sectors of economic activity at risk for the acquisition of these zoonoses. METHODS: A rapid review of the scientific literature was conducted. Databases on the Ovid and EBSCO research platforms were searched for articles published between 1995 and 2018, in English and French, on 14 zoonoses (campylobacteriosis, cryptosporidiosis, verocytotoxigenic Escherichia coli, giardiasis, listeriosis, salmonellosis, Eastern equine encephalitis, Lyme disease, West Nile virus, food botulism, Q fever, avian and swine influenza, rabies, hantavirus pulmonary syndrome) and occupational health. The literature search retrieved 12,558 articles and, after elimination of duplicates, 6,838 articles were evaluated based on the title and the abstract. Eligible articles had to address both concepts of the research issue (prioritized zoonoses and worker health). Of the 621 articles deemed eligible, 110 were selected following their full reading. RESULTS: Of the diseases under study, enteric zoonoses were the most frequently reported. Agriculture, including veterinary services, public administration services and medical and social services were the sectors most frequently identified in the literature. CONCLUSION: The results of our study will support public health authorities and decision-makers in targeting those sectors and occupations that are particularly at risk for the acquisition of zoonoses. Doing so will ultimately optimize the public health practices of those responsible for the health of workers.
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Though there is overlap, plant responses to caterpillar herbivory show distinct variations from mechanical wounding. In particular, effectors in caterpillar oral secretions modify wound-associated plant responses. Previous studies have focused on transcriptional and protein abundance differences in response to caterpillar herbivory. This study investigated Spodoptera exigua caterpillar-specific post-translational modification of Arabidopsis thaliana soluble leaf proteins by liquid chromatography/electrospray ionization/mass spectroscopy/mass spectroscopy (LC/ESI/MS/MS). Given that caterpillar labial saliva contains oxidoreductases, such as glucose oxidase, particular attention was paid to redox-associated modifications, such as the oxidation of protein cysteine residues. Caterpillar- and saliva-specific protein modifications were observed. Differential phosphorylation of the jasmonic acid biosynthetic enzyme, lipoxygenase 2, and a chaperonin protein is seen in plants fed upon by caterpillars with intact salivary secretions compared to herbivory by larvae with impaired labial salivary secretions. Often a systemic suppression of photosynthesis is associated with caterpillar herbivory. Of the five proteins modified in a caterpillar-specific manner (a transcription repressor, a DNA-repair enzyme, PS I P700, Rubisco and Rubisco activase), three are associated with photosynthesis. Oxidative modifications are observed, such as caterpillar-specific denitrosylation of Rubisco activase and chaperonin, cysteine oxidation of Rubisco, DNA-repair enzyme, and chaperonin and caterpillar-specific 4-oxo-2-nonenal modification of the DNA-repair enzyme.
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Proteínas de Arabidopsis/metabolismo , Processamento de Proteína Pós-Traducional/fisiologia , Spodoptera/fisiologia , Animais , Proteínas de Arabidopsis/análise , Larva , Espectrometria de Massas , Oxirredução , Folhas de Planta/química , Proteínas de Plantas/metabolismo , Proteômica/métodos , Saliva/enzimologiaRESUMO
Nicotiana benthamiana plants were agroinoculated with an infectious cDNA clone of Turnip mosaic virus (TuMV) that was engineered to express a fluorescent protein (green fluorescent protein [GFP] or mCherry) fused to the viral 6K2 protein known to induce vesicle formation. Cytoplasmic fluorescent discrete protein structures were observed in infected cells, corresponding to the vesicles containing the viral RNA replication complex. The vesicles were motile and aligned with microfilaments. Intracellular movement of the vesicles was inhibited when cells were infiltrated with latrunculin B, an inhibitor of microfilament polymerization. It was also observed that viral accumulation in the presence of this drug was reduced. These data indicate that microfilaments are used for vesicle movement and are necessary for virus production. Biogenesis of the vesicles was further investigated by infecting cells with two recombinant TuMV strains: one expressed 6K2GFP and the other expressed 6K2mCherry. Green- and red-only vesicles were observed within the same cell, suggesting that each vesicle originated from a single viral genome. There were also vesicles that exhibited sectors of green, red, or yellow fluorescence, an indication that fusion among individual vesicles is possible. Protoplasts derived from TuMV-infected N. benthamiana leaves were isolated. Using immunofluorescence staining and confocal microscopy, viral RNA synthesis sites were visualized as punctate structures distributed throughout the cytoplasm. The viral proteins VPg-Pro, RNA-dependent RNA polymerase, and cytoplasmic inclusion protein (helicase) and host translation factors were found to be associated with these structures. A single-genome origin and presence of protein synthetic machinery components suggest that translation of viral RNA is taking place within the vesicle.
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Brassica/virologia , Genoma Viral , Potyvirus/ultraestrutura , RNA Viral/metabolismo , Vesículas Transportadoras/metabolismo , Replicação Viral , Citoesqueleto de Actina/fisiologia , Citoesqueleto de Actina/ultraestrutura , Proteínas de Fluorescência Verde/genética , Proteínas de Fluorescência Verde/metabolismo , Microscopia Confocal , Potyvirus/genética , Potyvirus/metabolismo , Nicotiana/virologia , Vesículas Transportadoras/fisiologiaRESUMO
The aim of this study was to describe the epidemiology of human cryptosporidiosis in Québec from 2016 to 2017 and to identify possible exposures associated with the disease, and the dominant Cryptosporidium species in circulation. A descriptive analysis was performed on data collected from the provincial notifiable infectious diseases registry and the epidemiological investigation. Fecal sample were sent to the Laboratoire de santé publique du Québec for molecular characterization. In Québec, from January 1, 2016 to December 31, 2017, a total of 201 confirmed cases of cryptosporidiosis were notified. A peak in the number of reported cases was observed at the end of the summer. The regional public health department with the highest adjusted incidence rate for sex and age group for both years was that of Nunavik, in the north of Québec. A higher average annual incidence rate was observed for females between the ages of 20 to 34 years compared to males. Overall, for both males and females the distribution appeared to be bimodal with a first peak in children younger than five years old and a second peak in adults from 20 to 30 years of age. Molecular characterization showed that 23% (11/47) of cases were infected with C. hominis while 74% (35/47) were infected with C. parvum. Meanwhile, subtyping results identified by gp60 sequencing, show that all C. parvum subtypes belonged to the IIa family, whereas the subtypes for C. hominis belonged to the Ia, Ib, and Id families. Finally, the epidemiological investigation showed that diarrhea was the most common reported symptom with 99% (72/73) of investigated cases having experienced it. This first brief epidemiological portrait of cryptosporidiosis in Québec has allowed for the description, both at the provincial and regional level, of the populations that could be particularly vulnerable to the disease.
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Criptosporidiose/epidemiologia , Adulto , Canadá/epidemiologia , Feminino , Humanos , Incidência , Masculino , Saúde Pública/estatística & dados numéricos , Quebeque/epidemiologia , Adulto JovemRESUMO
BACKGROUND: Sm16, also known as SPO-1 and SmSLP, is a low molecular weight protein (~16kDa) secreted by the digenean trematode parasite Schistosoma mansoni, one of the main causative agents of human schistosomiasis. The molecule is secreted from the acetabular gland of the cercariae during skin invasion and is believed to perform an immune-suppressive function to protect the invading parasite from innate immune cell attack. METHODOLOGY/PRINCIPAL FINDINGS: We show that Sm16 homologues of the Schistosomatoidea family are phylogenetically related to the helminth defence molecule (HDM) family of immunomodulatory peptides first described in Fasciola hepatica. Interrogation of 69 helminths genomes demonstrates that HDMs are exclusive to trematode species. Structural analyses of Sm16 shows that it consists predominantly of an amphipathic alpha-helix, much like other HDMs. In S. mansoni, Sm16 is highly expressed in the cercariae and eggs but not in adult worms, suggesting that the molecule is of importance not only during skin invasion but also in the pro-inflammatory response to eggs in the liver tissues. Recombinant Sm16 and a synthetic form, Sm16 (34-117), bind to macrophages and are internalised into the endosomal/lysosomal system. Sm16 (34-117) elicited a weak pro-inflammatory response in macrophages in vitro but also suppressed the production of bacterial lipopolysaccharide (LPS)-induced inflammatory cytokines. Evaluation of the transcriptome of human macrophages treated with a synthetic Sm16 (34-117) demonstrates that the peptide exerts significant immunomodulatory effects alone, as well as in the presence of LPS. Pathways most significantly influenced by Sm16 (34-117) were those involving transcription factors peroxisome proliferator-activated receptor (PPAR) and liver X receptors/retinoid X receptor (LXR/RXR) which are intricately involved in regulating the cellular metabolism of macrophages (fatty acid, cholesterol and glucose homeostasis) and are central to inflammatory responses. CONCLUSIONS/SIGNIFICANCE: These results offer new insights into the structure and function of a well-known immunomodulatory molecule, Sm16, and places it within a wider family of trematode-specific small molecule HDM immune-modulators with immuno-biotherapeutic possibilities.
Assuntos
Antígenos de Helmintos/metabolismo , Proteínas de Helminto/metabolismo , Schistosoma mansoni/metabolismo , Animais , Anticorpos Anti-Helmínticos , Antígenos de Helmintos/genética , Antígenos de Helmintos/imunologia , Células da Medula Óssea , Linhagem Celular Tumoral , Regulação da Expressão Gênica , Proteínas de Helminto/genética , Humanos , Macrófagos/parasitologia , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos C57BL , Óvulo , Filogenia , Transporte ProteicoRESUMO
BACKGROUND: Lyme disease is an emerging vector-borne zoonotic disease of increasing public health importance in Canada. As part of its mandate, the Canadian Lyme Disease Research Network (CLyDRN) launched a pan-Canadian sentinel surveillance initiative, the Canadian Lyme Sentinel Network (CaLSeN), in 2019. OBJECTIVES: To create a standardized, national sentinel surveillance network providing a real-time portrait of the evolving environmental risk of Lyme disease in each province. METHODS: A multicriteria decision analysis (MCDA) approach was used in the selection of sentinel regions. Within each sentinel region, a systematic drag sampling protocol was performed in selected sampling sites. Ticks collected during these active surveillance visits were identified to species, and Ixodes spp. ticks were tested for infection with Borrelia burgdorferi, Borrelia miyamotoi, Anaplasma phagocytophilum, Babesia microti and Powassan virus. RESULTS: In 2019, a total of 567 Ixodes spp. ticks (I. scapularis [n=550]; I. pacificus [n=10]; and I. angustus [n=7]) were collected in seven provinces: British Columbia, Manitoba, Ontario, Québec, New Brunswick, Nova Scotia and Prince Edward Island. The highest mean tick densities (nymphs/100 m2) were found in sentinel regions of Lunenburg (0.45), Montréal (0.43) and Granby (0.38). Overall, the Borrelia burgdorferi prevalence in ticks was 25.2% (0%-45.0%). One I. angustus nymph from British Columbia was positive for Babesia microti, a first for the province. The deer tick lineage of Powassan virus was detected in one adult I. scapularis in Nova Scotia. CONCLUSION: CaLSeN provides the first coordinated national active surveillance initiative for tick-borne disease in Canada. Through multidisciplinary collaborations between experts in each province, the pilot year was successful in establishing a baseline for Lyme disease risk across the country, allowing future trends to be detected and studied.
RESUMO
Since its detection in Canada in the early 1990s, Ixodes scapularis, the primary tick vector of Lyme disease in eastern North America, has continued to expand northward. Estimates of the tick's broad-scale distribution are useful for tracking the extent of the Lyme disease risk zone; however, tick distribution may vary widely within this zone. Here, we investigated I. scapularis nymph distribution at three spatial scales across the Lyme disease emergence zone in southern Quebec, Canada. We collected ticks and compared the nymph densities among different woodlands and different plots and transects within the same woodland. Hot spot analysis highlighted significant nymph clustering at each spatial scale. In regression models, nymph abundance was associated with litter depth, humidity, and elevation, which contribute to a suitable habitat for ticks, but also with the distance from the trail and the type of trail, which could be linked to host distribution and human disturbance. Accounting for this heterogeneous nymph distribution at a fine spatial scale could help improve Lyme disease management strategies but also help people to understand the risk variation around them and to adopt appropriate behaviors, such as staying on the trail in infested parks to limit their exposure to the vector and associated pathogens.