RESUMO
Echinococcus granulosus sensu lato is a platyhelminth parasite and the etiological cause of cystic echinococcosis (CE), a zoonotic and neglected disease that infects animals and humans worldwide. As a part of the biological arsenal of the parasite, cathepsin L proteases are a group of proteins that are believed to be essential for parasite penetration, immune evasion, and establishment in the tissues of the host. In this work, we have cloned and sequenced a new putative cathepsin L protease from Echinococcus canadensis (EcCLP1). The bioinformatic analysis suggests that EcCLP1 could be synthesized as a zymogen and activated after proteolytic cleavage. The multiple sequence alignment with other cathepsin proteases reveals important functional conserved features like a conserved active site, an N-linked glycosylation residue, a catalytic triad, an oxyanion hole, and three putative disulfide bonds. The phylogenetic analysis suggests that EcCLP1 could indeed be a cathepsin L cysteine protease from clade 1 as it grouped with cathepsins from other species in this clade. Modeling studies suggest that EcCLP1 has two domains forming a cleft where the active site is located and an occluding role for the propeptide. The transcriptomic analysis reveals different levels of cathepsin transcript expression along the different stages of the parasite life cycle. The whole-mount immunohistochemistry shows an interesting superficial punctate pattern of staining which suggests a secretory pattern of expression. The putative cathepsin L protease characterized here may represent an interesting tool for diagnostic purposes, vaccine design, or a new pharmacological target for antiparasitic intervention.
Title: Caractérisation moléculaire d'EcCLP1, une nouvelle protéase putative de type cathepsine L d'Echinococcus canadensis. Abstract: Echinococcus granulosus sensu lato est un Plathelminthe parasite et la cause étiologique de l'échinococcose kystique (EK), une maladie zoonotique et négligée qui infecte les animaux et les humains dans le monde entier. En tant que partie de l'arsenal biologique du parasite, les protéases de type cathepsine L sont un groupe de protéines considérées comme essentielles à la pénétration du parasite, l'évasion immunitaire et son établissement dans les tissus de l'hôte. Dans ce travail, nous avons cloné et séquencé une nouvelle protéase putative de type cathepsine L d'Echinococcus canadensis (EcCLP1). L'analyse bioinformatique suggère qu'EcCLP1 pourrait être synthétisée sous forme de zymogène et activée après clivage protéolytique. L'alignement de séquences multiples avec d'autres protéases de type cathepsine révèle d'importantes caractéristiques fonctionnelles conservées telles qu'un site actif conservé, un résidu de glycosylation lié à N, une triade catalytique, un trou oxyanion et trois liaisons disulfure putatives. L'analyse phylogénétique suggère qu'EcCLP1 pourrait en effet être une protéase de type cathepsine L du clade 1 car elle se regroupe avec les cathepsines d'autres espèces de ce clade. Les études de modélisation suggèrent qu'EcCLP1 possède deux domaines formant une fente où se trouve le site actif et un rôle d'occlusion pour le propeptide. L'analyse transcriptomique révèle différents niveaux d'expression du transcrit de la cathepsine au cours des différentes étapes du cycle de vie du parasite. L'immunohistochimie de montages entiers montre un intéressant motif de coloration ponctuée superficielle qui suggère un modèle d'expression sécrétoire. La protéase putative de type cathepsine L caractérisée ici peut représenter un outil intéressant à des fins de diagnostic, de conception de vaccins ou une nouvelle cible pharmacologique pour une intervention antiparasitaire.
Assuntos
Sequência de Aminoácidos , Catepsina L , Echinococcus , Filogenia , Animais , Catepsina L/genética , Echinococcus/enzimologia , Echinococcus/genética , Echinococcus/classificação , Alinhamento de Sequência , Clonagem Molecular , Proteínas de Helminto/genética , Proteínas de Helminto/química , Estágios do Ciclo de Vida , Equinococose/parasitologia , Domínio Catalítico , Perfilação da Expressão GênicaRESUMO
OBJECTIVE: To investigate the feasibility of constructing the risk index of Echinococcus infection based on the classification of echinococcosis lesions, so as to provide insights into the management of echinococcosis. METHODS: The imaging data of echinococcosis cases were collected from epidemiological surveys of echinococcosis in China from 2012 to 2016, and the detection of incident echinococcosis cases was captured from the annual echinococcosis prevention and control reports across provinces (autonomous regions) and Xinjiang Production and Construction Corps in China from 2017 to 2022. After echinococcosis lesions were classified, a risk index of Echinococcus infection was constructed based on the principle of discrete distribution marginal probability and multi-group classification data tests. The correlation between the risk index of Echinococcus infection and the detection of incident echinococcosis cases was evaluated in the provinces (autonomous regions and corps) from 2017 to 2022, and the correlations between the short and medium-term risk indices and between the medium and long-term risk indices of Echinococcus infection were examined using a univariate linear regression model. RESULTS: A total of 4 014 echinococcosis cases in China from 2012 to 2016 were included in this study. The short-, medium- and long-term risk indices of E. granulosus infection varied in echinococcosis-endemic provinces (autonomous regions and corps) of China (χ2 = 4.12 to 708.65, all P values < 0.05), with high short- (0.058), medium- (0.137) and long-term risk indices (0.104) in Tibet Autonomous Region, and the short-, medium- and long-term risk indices of E. multilocularis infection varied in echinococcosis-endemic provinces (autonomous regions and corps) of China (χ2 = 6.74 to 122.60, all P values < 0.05), with a high short-term risk index in Sichuan Province (0.016) and high medium- (0.009) and long-term risk indices in Qinghai Province (0.018). There were no significant correlations between the risk index of E. granulosus infection and the detection of incident cystic echinococcosis cases during the study period (t = -0.518 to 2.265, all P values > 0.05), and strong correlations were found between the risk indices of E. multilocularis infection and the detection of incident alveolar echinococcosis cases (including mixed type) in 2018, 2020, 2021, 2022, during the period from 2017 through 2020, from 2017 through 2021, from 2017 through 2022 (all r values > 0.7, t = 2.521 to 3.692, all P values < 0.05). Linear regression models were established between the risk index of E. multilocular infection and the detection of alveolar echinococcosis cases (including mixed type), and the models were all statistically significant (b = 0.214 to 2.168, t = 2.458 to 3.692, F = 6.044 to 13.629, all P values < 0.05). The regression coefficients for the correlations between the medium- and short-term, and between the long- and medium-term risk indices of E. granulosus infection were 2.339 and 0.765, and the regression coefficients for the correlations between the medium- and short-term, and between the long- and medium-term risk indices of E. multilocular infection were 0.280 and 1.842, with statistical significance seen in both the regression coefficients and regression models (t = 16.479 to 197.304, F = 271.570 to 38 928.860, all P values < 0.05). CONCLUSIONS: The risk index of Echinococcus infection has been successfully established based on the classification of echinococcosis lesions, which may provide insights into the prevention and control, prediction, diagnosis and treatment, and classified management of echinococcosis.
Assuntos
Equinococose , Equinococose/epidemiologia , Equinococose/parasitologia , Equinococose/diagnóstico , Humanos , China/epidemiologia , Echinococcus/isolamento & purificação , Echinococcus/fisiologia , Echinococcus/classificação , Fatores de Risco , AnimaisRESUMO
OBJECTIVE: To investigate the prevalence of Echinococcus infections in small rodents around human residential areas in Yushu City, Qinghai Province in 2023, so as to provide insights into precision echinococcosis control. METHODS: One or two quadrats, each measuring 50 m × 50 m, were randomly assigned in Shanglaxiu Township and Longbao Township, Yushu City, Qinghai Province on June 2023, respectively, and 300 plate-type mouse traps, each measuring 12.0 cm × 6.5 cm, were assigned in each quadrat. Small rodents were captured during the period between 10 : 00 and 18 : 00 each day for 4 days. Then, all captured small rodents were identified and dissected, and liver specimens with suspected Echinococcus infections were subjected to pathological examinations. The Echinococcus cytochrome c oxidase 1 (cox1) gene was amplified using PCR assay, and the sequence of the amplified product was aligned to that was recorded in the GenBank to characterize the parasite species. In addition, a phylogenetic tree of Echinococcus was generated based on the cox1 gene sequence using the neighbor-joining method. RESULTS: A total of 236 small rodents were captured in Shanglaxiu and Longbao townships, Yushu City, including 65 Qinghai voles and 51 plateau pikas in Shanglaxiu Township, and 62 Qinghai voles and 58 plateau pikas in Longbao Township, and there was no significant difference in the constituent ratio of small rodents between the two townships (χ2 = 0.294, P > 0.05). Seven plateau pikas and 12 Qinghai voles were suspected to be infected with Echinococcus by dissection, and pathological examinations showed unclear structure of hepatic lobules and disordered hepatocyte arrangement in livers of small rodents suspected of Echinococcus infections. PCR assay identified E. shiquicus DNA in 7 Qinghai voles, which were all captured from Shanglaxiu Township. Phylogenetic analysis showed that the cox1 gene sequence of Echinococcus in small rodents was highly homologous to the E. shiquicus cox1 gene sequence reported previously. CONCLUSIONS: Plateau pika and Qinghai vole were predominant small rodents around human residential areas in Yushu City, Qinghai Province in 2023, and E. shiquicus infection was detected in Qinghai voles.
Assuntos
Equinococose , Echinococcus , Filogenia , Roedores , Animais , Equinococose/epidemiologia , Equinococose/veterinária , Equinococose/parasitologia , China/epidemiologia , Echinococcus/genética , Echinococcus/isolamento & purificação , Echinococcus/classificação , Roedores/parasitologia , Prevalência , HumanosRESUMO
Echinococcus granulosus sensu lato (s.l.) is a species complex with the potential to cause cystic echinococcosis (CE). Contact with the feces of domestic dogs (Canis familiaris) fed with raw viscera of intermediate livestock hosts is a risk factor for this infection in the southern region of Brazil. Although the region has been considered endemic to CE for many years, molecular data regarding the species of the complex causing CE in humans are scarce. This study aimed to perform a molecular analysis of the biological fluid from a human liver cyst to investigate the species responsible for CE. Genetic material obtained from the hydatid fluid of a hepatic cyst from a human with CE was subjected to PCR to amplify mitochondrial and nuclear DNA sequences. The phylogenetic analysis confirmed the human infection by Echinococcus canadensis G7 in the state of Paraná, Brazil. This is the first molecular record of E. canadensis G7 infecting a human in Brazil, and it is important to reiterate the risk of human CE caused by this species in South America, as reported by a previous study in Patagonia, Argentina. From the epidemiological point of view, this finding is of great relevance for the southern region of Brazil, since this parasite has previously only been detected in pigs in the state of Rio Grande do Sul, neighboring Paraná. The finding points to the importance of this identification in the molecular epidemiology of E. granulosus s.l., especially in South America.
Assuntos
DNA de Helmintos , Echinococcus , Filogenia , Animais , Brasil/epidemiologia , Echinococcus/genética , Echinococcus/classificação , Echinococcus/isolamento & purificação , Humanos , DNA de Helmintos/genética , Equinococose/veterinária , Equinococose/parasitologia , Equinococose/epidemiologia , Análise de Sequência de DNA , Reação em Cadeia da Polimerase , DNA Mitocondrial/genética , MasculinoRESUMO
Echinococcosis is a zoonotic disease with great significance to public health, and appropriate detection and control strategies should be adopted to mitigate its impact. Most cases of echinococcosis are believed to be transmitted by the consumption of food and/or water contaminated with canid stool containing Echinococcus spp. eggs. Studies assessing Echinococcus multilocularis, Echinococcus granulosus sensu stricto, and Echinococcus shiquicus coinfection from contaminated water-derived, soil-derived, and food-borne samples are scarce, which may be due to the lack of optimized laboratory detection methods. The present study aimed to develop and evaluate a novel triplex TaqMan-minor groove binder probe for real-time polymerase chain reaction (rtPCR) to simultaneously detect the 3 Echinococcus spp. mentioned above from canid fecal samples in the Qinghai-Tibetan Plateau area (QTPA). The efficiency and linearity of each signal channel in the triplex rtPCR assay were within acceptable limits for the range of concentrations tested. Furthermore, the method was shown to have good repeatability (standard deviation ≤0.32 cycle threshold), and the limit of detection was estimated to be 10 copies plasmid/µl reaction. In summary, the evaluation of the present method shows that the newly developed triplex rtPCR assay is a highly specific, precise, consistent, and stable method that could be used in epidemiological investigations of echinococcosis.
Assuntos
Canidae/parasitologia , Doenças do Cão/parasitologia , Equinococose/veterinária , Echinococcus/isolamento & purificação , Fezes/parasitologia , Reação em Cadeia da Polimerase Multiplex/veterinária , Animais , Biologia Computacional/normas , DNA de Helmintos/isolamento & purificação , Cães , Equinococose/parasitologia , Echinococcus/classificação , Echinococcus/genética , Raposas/parasitologia , Limite de Detecção , Reação em Cadeia da Polimerase Multiplex/métodos , Reação em Cadeia da Polimerase Multiplex/normas , Reprodutibilidade dos Testes , Sensibilidade e Especificidade , Solo/parasitologiaRESUMO
Echinococcosis is a zoonotic parasitic disease that is highly endemic to the Qinghai province of China. Limited data are available on the prevalence of the causal pathogen, Echinococcus spp., in definitive hosts in this region. Thus, the aim of this study was to evaluate the prevalence of Echinococcus spp. in wild foxes and stray dogs in Qinghai province. Five hundred and twenty-eight feces from wild foxes and 277 from stray dogs were collected from 11 counties in the Golog, Yushu, and Haixi prefectures and screened for Echinococcus spp. using copro-DNA polymerase chain reaction (PCR). In total, 5.5% of wild foxes and 15.2% of stray dogs tested positive for Echinococcus spp. The prevalence rates of Echinococcus spp. in wild foxes in Golog, Yushu, and Haixi were 7.3%, 5.2%, and 1.9%, respectively. In stray dogs, these rates were 13.3%, 17.3%, and 0%, respectively. Sequencing analysis determined that Echinococcus multilocularis was the most prevalent species, occurring in 4.0% and 12.6% of wild foxes and stray dogs, respectively. Echinococcus shiquicus was observed in 1.5% of wild foxes and 0.7% of stray dogs. Echinococcus granulosus was observed only in wild dogs, with a prevalence rate of 1.8%. To our knowledge, this is the first report on the prevalence of E. shiquicus in dogs in Qinghai province. The current results improve our understanding of the transmission and dissemination of human echinococcosis and suggest that exposure to the eggs of E. multilocularis harbored by wild foxes and stray dogs may pose a great risk of alveolar echinococcosis to humans in Qinghai province.
Assuntos
Animais Selvagens/parasitologia , Equinococose/epidemiologia , Echinococcus/genética , Raposas/parasitologia , Animais , China/epidemiologia , DNA de Helmintos/genética , Doenças do Cão/epidemiologia , Doenças do Cão/parasitologia , Cães , Equinococose/parasitologia , Equinococose/transmissão , Echinococcus/classificação , Fezes/parasitologia , Prevalência , Zoonoses/epidemiologia , Zoonoses/parasitologiaRESUMO
Echinococcosis, caused by genus Echinococcus, is the most pathogenic zoonotic parasitic disease in the world. In Tibet of the People's Republic of China, echinococcosis refers principally to two types of severe zoonosis, cystic echinococcosis (CE) and alveolar echinococcosis (AE), which place a serious burden on public health and economy in the local community. However, research on the spatial epidemiology of echinococcosis remains inadequate in Tibet, China. Based on the recorded human echinococcosis data, maps of the spatial distribution of human CE and AE prevalence in Tibet were produced at city level and county level respectively, which show that the prevalence of echinococcosis in northern and western Tibet was much higher than that in other regions. We employ a geographical detector to explore the influencing factors for causing CE and AE while sorting information on the maps of disease prevalence and environment factors (e.g. terrain, population, and yak population). The results of our analysis showed that biological factors have the most impact on the prevalence of echinococcosis, of which the yak population contributes the most for CE, while the dog population contributes the most for AE. In addition, the interaction between various factors, as we found out, might further explain the disease prevalence, which indicated that the echinococcosis prevalence is not simply affected by one single factor, but by multiple factors that are correlated with each other complicatedly. Our results will provide an important reference for the evaluation of the echinococcosis risk, control projects, and prevention programs in Tibet.
Assuntos
Equinococose/parasitologia , Equinococose/veterinária , Echinococcus/isolamento & purificação , Animais , Bovinos , Doenças dos Bovinos/epidemiologia , Doenças dos Bovinos/parasitologia , Doenças do Cão/epidemiologia , Doenças do Cão/parasitologia , Cães , Equinococose/epidemiologia , Echinococcus/classificação , Echinococcus/genética , Humanos , Prevalência , Tibet/epidemiologia , Zoonoses/epidemiologia , Zoonoses/parasitologiaRESUMO
The paraphyletic group Echinococcus granulosus sensu lato is comprised of parasitic tapeworms of wild and domestic canids such as wolves (Canis lupus) and coyotes (Canis latrans), which serve as definitive hosts, and ungulates, which are the intermediate hosts. Members of this tapeworm group are characterized by both cosmopolitan distribution and zoonotic disease potential. This survey (conducted from 2012 through 2017) was designed to provide insight into the prevalence and distribution of this parasite in wild canids in Wyoming. Echinococcus sp. infections were documented in 14 of 22 gray wolves (63.6%), 1 of 182 coyotes (0.55%) and 0 of 5 red foxes (Vulpes fulva). Echinococcus granulosus s. l. was confirmed in 4 of these 14 specimens obtained from wolves with two parasite specimens corresponding morphologically with E. canadensis (G8/G10). These results suggest that wolves serve as the major definitive host of E. granulosus s. l. in Wyoming, while coyotes do not play an equivalent role. Limited sample size precludes evaluation of the importance of the red fox as a favorable definitive host. Whereas this study documents the occurrence of E. granulosus s. l. in Wyoming, the zoonotic disease risk does not appear to be high. Education remains the key to disease prevention, coupled with good hygienic practices by humans and anthelmintic treatment of domestic dogs exhibiting elevated risk of exposure.
Assuntos
Coiotes/parasitologia , Equinococose/veterinária , Echinococcus granulosus , Raposas/parasitologia , Lobos/parasitologia , Animais , Animais Selvagens/parasitologia , Equinococose/epidemiologia , Equinococose/parasitologia , Echinococcus/classificação , Echinococcus granulosus/classificação , Feminino , Masculino , Wyoming/epidemiologia , ZoonosesRESUMO
BACKGROUND: Cystic or alveolar echinococcosis caused by the larval stages of Echinococcus spp. is a very severe zoonotic helminth infection. Echinococcus shiquicus is a newly discovered species that has only been reported in the Qinghai and Sichuan provinces of the Qinghai-Tibet plateau, China where, to date, it has only been confirmed in Tibetan foxes and wild small mammal populations of the Tibetan plateau. Information on its genetic and evolutionary diversity is scanty. The aim of this study was to investigate the prevalence of E. shiquicus in plateau pikas (Ochotona curzoniae), a known intermediate host, and to determine the genetic variation and phylogenetic relationship of the E. shiquicus population in the Tibet region of China based on mitochondrial DNA. METHODS: Echinococcus shiquicus samples were collected from Damxung and Nyêmo counties (located in Tibet Autonomous Region, China). The mitochondrial cox1 and nad1 gene sequences were analyzed, and the genetic diversity and epidemiology of E. shiquicus in the region were discussed based on the results. RESULTS: The prevalence of E. shiquicus in pikas in Damxung and Nyêmo counties was 3.95% (6/152) and 6.98% (9/129), respectively. In combination with previous public sequence data, the haplotype analysis revealed 12 haplotypes (H) characterized by two distinct clusters (I and II), and a sequence distance of 99.1-99.9% from the reference haplotype (H1). The diversity and neutrality indices for the entire E. shiquicus populations were: haplotype diversity (Hd) ± standard deviation (SD) 0.862 ± 0.035; nucleotide diversity (Hd ± SD) 0.0056 ± 0.0003; Tajima's D 0.876 (P > 0.05); and Fu's F 6.000 (P > 0.05). CONCLUSIONS: This was the first analysis of the newly discovered E. shiquicus in plateau pikas in the Tibet Autonomous Region of China. The neutrality indices suggest a deficiency of alleles, indicative of a recent population bottleneck.
Assuntos
Equinococose/epidemiologia , Equinococose/veterinária , Echinococcus/genética , Variação Genética , Lagomorpha/parasitologia , Filogenia , Animais , China/epidemiologia , Equinococose/parasitologia , Echinococcus/classificação , Echinococcus/isolamento & purificação , Raposas/parasitologia , Genótipo , Prevalência , Tibet/epidemiologiaRESUMO
Cystic echinococcosis (CE) is endemic in many parts of sub-Saharan Africa. In contrast to the eastern part of the continent, very little data exists on the current disease situation in southern Africa including Zambia. This study determined frequency and species identity of Echinococcus spp. circulating in livestock and dogs in the Western Province of Zambia. Cysts were collected in slaughterhouses at meat inspection (cattle) and during examination of home slaughtered pigs, while dog faecal samples were collected per-rectum and examined microscopically for the presence of taeniid eggs. Individual taeniid eggs from faecal samples and individual protoscoleces from cysts were genotyped by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and/or sequencing of the NADH-dehydrogenase subunit 1 (nad1) and cytochrome C oxidase 1 (cox1) gene. Fifty-four of 2000 cattle (2.7%) were found infected with a total of 65 cysts, predominantly fertile lungs cysts; all cysts were identified as Echinococcus ortleppi. Two out of 52 home-slaughtered pigs (3.8%) were infected with a fertile lung cyst each; both cysts were also identified as E. ortleppi. Microscopic examination revealed 10/289 dog faecal samples to contain taeniid eggs, of which four samples (two each) contained Echinococcus canadensis (G6/7) or Taenia hydatigena, respectively. This is the first insight in the Echinococcus species circulating in Zambia providing premises for further studies into transmission dynamics of CE in the southern African region.
Assuntos
Equinococose/veterinária , Echinococcus/classificação , Animais , Animais Domésticos , Equinococose/epidemiologia , Equinococose/parasitologia , Echinococcus/genética , Fezes , Genótipo , Reação em Cadeia da Polimerase/veterinária , Polimorfismo de Fragmento de Restrição , Zâmbia/epidemiologiaRESUMO
Cystic echinococcosis (CE), a worldwide zoonosis, is highly prevalent in Africa particularly in northern and eastern Africa where data are more abundant than other regions. However, harmonization of available data through systematic review and meta-analysis may foster improved transboundary cooperation for the control of CE in Africa. Using the Preferred Reporting Items for Systematic Reviews and Meta-analyses guidelines, research articles (from 2000 to 2019) were retrieved from ScienceDirect, PubMed, African Journals OnLine and Google Scholar databases. A total of 98 studies of 806,624 animals from 13 countries comprising 264,016 goats, 247,326 sheep, 251,106 cattle, 28,314 camels, 4,764 buffaloes, 2,920 equids, 1,966 pigs, 408 wild boars and 50 Norway rats were available for systematic review and meta-analysis of pooled prevalence including 5,048 dogs, 345 lions, 220 hyenas, 94 wolves and 47 jackals/foxes analysed for Echinococcus infection. In total, 46,869 animals were infected and pooled prevalence of CE in intermediate hosts was highest in camels (17.1%; 95% CI: 12.1-22.8) and lowest in pigs (0.3%; 95% CI: 0.1-0.6). Results also showed uneven species/genotype distribution across the continent such that Echinococcus granulosus sensu stricto (G1, G3) constituted 74.45% of the total isolates from East Africa, E. canadensis (G6/7) accounted for 60.3% and 97.4% in North and West Africa, respectively, while 81.3% of E. ortleppi (G5) were recorded for southern Africa. The comparatively higher prevalence estimates for eastern and northern Africa than other regions indicate where efforts on CE management should now be given greater attention in Africa. Additionally, this study also advocates for better cooperation between countries within the same sub-region and the establishment of joint CE control programmes.
Assuntos
Animais Domésticos/parasitologia , Animais Selvagens/parasitologia , Equinococose/veterinária , Echinococcus/classificação , Echinococcus/isolamento & purificação , África/epidemiologia , Animais , Equinococose/epidemiologia , Equinococose/parasitologia , Echinococcus/genética , Echinococcus granulosus/classificação , Echinococcus granulosus/genética , Genótipo , PrevalênciaRESUMO
Cystic echinococcosis (CE) is a zoonotic infection and could lead to significant public health problems. The genetic diversity of CE includes five species: E. granulosus sensu stricto (s.s.) (G1-G3), Echinococcus equinus (G4), Echinococcus ortleppi (G5), Echinococcus canadensis genotypic cluster (G6, G7, G8 and G10, with the doubtful G9) and the Echinococcus felidis (lion strain). The species are important in epidemiology, pathology, control, prevention measures and vaccine/drug designs. The aim of the present study was to determine the E. granulosus genotypes in humans in the Van province in east of Turkey. In total, 102 echinococcal cysts were collected from operated patients. Genomic analyses were conducted with PCR-RFLP of the rDNA internal transcribed spacer 1 (ITS1) fragment and partial PCR sequencing of the cytochrome c oxidase subunit 1 (cox1) mitochondrial DNA gene region. In total, DNAs of 96 isolates could be extracted, unfortunately six extractions failed. The PCR-RFLP analysis findings were identical in all isolates. Two bands were observed at approximately 300 bp and 600 bp. All profiles corresponded to the G1-G3 strain. Also, 446 bp amplified gene regions were observed for cox1. Out of 20 samples, alignment of 16 sequences exhibited a total identification (100%) of granulosus sensu stricto (G1/G3). Of 16 samples, 8 were obtained in the lung and 12 were obtained in the liver; 8 belonged to male and 12 belonged to female patients. Other four samples exhibited one nucleotide substitution at different positions. Four samples had one nucleotide substitution at different positions. We detected single nucleotide variations in TRH1, TRH67, TRH85 and TRH89 isolates at the positions C240T; G330T; G211A and T157C, respectively. In conclusion, the present study was the first comprehensive molecular investigation on genetic characterization of human CE isolates in Van region. The findings demonstrated that E. granulosus s.s. was the dominant species, which indicated that the sheep-dog cycle was the source in human infections. And, probably, it would be possible to describe these mutations as "Turkey" or "lung" variants. In addition to contributing molecular epidemiological data, the present results should be considered when designing and implementing E. granulosus control programs.
Assuntos
Equinococose Hepática/parasitologia , Equinococose Pulmonar/parasitologia , Echinococcus/genética , Animais , Echinococcus/classificação , Feminino , Humanos , MasculinoRESUMO
The parasitic species of the Echinococcus granulosus sensu lato (sl) complex are the causative agents of cystic echinococcosis in humans. The lifecycle of E. granulosus sl is essentially domestic, and is based on the consumption by dogs of hydatid cysts in viscera of livestock species. The aim of this study was to survey E. granulosus sensu lato in livestock in France. A 1-year national survey of E. granulosus sl in livestock at the slaughterhouse was organized in 2012 in France, with systematic molecular confirmation. The prevalence of E. granulosus ss nationally was 0.002% in sheep, mainly focused in the Alpine area, and 0.001% in cattle, with the distribution of cases throughout the country. Echinococcus canadensis G6/7 was observed only in Corsica in pigs, with a prevalence of nearly 1% in the island. A national prevalence of 0.0002% was estimated for E. ortleppi in cattle, due to seven cases distributed in two foci. The results of this survey are of particular interest because of the zoonotic risk associated with the presence of these parasite species, for which systematic control at the slaughterhouse should enable their elimination.
Assuntos
Equinococose/veterinária , Echinococcus granulosus/isolamento & purificação , Echinococcus/isolamento & purificação , Distribuição Animal , Animais , Bovinos , Doenças dos Bovinos/epidemiologia , Doenças dos Bovinos/parasitologia , Equinococose/epidemiologia , Equinococose/parasitologia , Echinococcus/classificação , Echinococcus/genética , Echinococcus granulosus/genética , França/epidemiologia , Doenças das Cabras/epidemiologia , Doenças das Cabras/parasitologia , Cabras , Doenças dos Cavalos/epidemiologia , Doenças dos Cavalos/parasitologia , Cavalos , Prevalência , Ovinos , Doenças dos Ovinos/epidemiologia , Doenças dos Ovinos/parasitologia , Carneiro Doméstico , Sus scrofa , Suínos , Doenças dos Suínos/epidemiologia , Doenças dos Suínos/parasitologiaRESUMO
BACKGROUND: Cystic echinococcosis (CE) in humans and livestock is caused by Echinococcus granulosus (sensu lato). In China where CE is endemic, a number of studies have shown that Echinococcus granulosus (sensu stricto) is majorly responsible for CE. However, E. canadensis (G6) which is the second leading cause of CE is now being detected in most parts of the country. In this study, the species diversity and genetic variation of Echinococcus granulosus (s.l.) in four counties in Tibet Autonomous Region of China were investigated. METHODS: Infection with Echinococcus granulosus (s.s.) in yaks and sheep was identified using NADH dehydrogenase subunit 1 and 5 (nad1 and nad5) mitochondrial genes while the genotype G6 of E. canadensis initially diagnosed with NADH dehydrogenase subunit 1 (nad1) was further confirmed by analysis of the complete mitochondrial genome and a phylogenetic network constructed based on the nad2 and nad5 genes. RESULTS: Out of 85 hydatid cyst samples collected from slaughtered sheep (n = 54) and yaks (n = 31), 83 were identified as E. granulosus (s.s.) G1 (n = 77), G3 (n = 6) and 2 were identified as E. canadensis G6. Analysis of the nad1/nad5 genes revealed 16/17 mutations with 9/14 parsimony informative sites resulting in 15/14 haplotypes, respectively. Haplotype diversity (Hd) and nucleotide diversity (π) of E. granulosus (s.s.) population were 0.650 and 0.00127 for nad1 and 0.782 and 0.00306 for nad5, respectively, with an overall negative Tajima's D and Fu's Fs. A low FST indicated no genetic difference between isolates from sheep and yaks. CONCLUSION: Pockets of infection with E. canadensis (G6, G7, G8 and G10) have been previously reported in sheep, goats, yaks and/or humans in different parts of China. While the G6 genotype has been previously reported in sheep in the Tibet Autonomous Region, the detection in a yak in the present study represents the first to the best of our knowledge. Therefore, we recommend future surveys and control efforts to comprehensively investigate other potential intermediate hosts for the prevalence and genetic diversity of the E. canadensis group (G6, G7, G8 and G10) across the country and their inclusion into the existing CE control programme.
Assuntos
Doenças dos Bovinos/parasitologia , DNA de Helmintos/genética , DNA Mitocondrial/genética , Equinococose/veterinária , Echinococcus/genética , Variação Genética , Doenças dos Ovinos/parasitologia , Animais , Bovinos , Equinococose/parasitologia , Echinococcus/classificação , Echinococcus/isolamento & purificação , Genótipo , Filogenia , Ovinos , TibetRESUMO
The diversity and importance of Echinococcus species in domesticated animals in Kazakhstan are poorly understood. In this study, 17 cysts of Echinococcus were collected from cattle and a further 17 cysts from sheep. DNA was extracted from the individual cysts and used for polymerase chain reaction amplification of mitochondrial subunit 1 of the cox1 and nadh1 gene. Amplicon sequencing results revealed the presence of Echinococcus granulosus sensu stricto G1 in 15 cattle and 15 sheep, and G3 genotype from two cattle. Echinococcus canadensis (G6/G7 strain) was found in two cysts originating from sheep.
Assuntos
DNA de Helmintos/genética , Equinococose/veterinária , Echinococcus/genética , Genótipo , Gado/parasitologia , Animais , Bovinos/parasitologia , Equinococose/parasitologia , Echinococcus/classificação , Echinococcus/isolamento & purificação , Cazaquistão , Ovinos/parasitologiaRESUMO
Cystic echinococcosis (CE) is a neglected zoonotic disease with considerable economic and public health burden worldwide, particularly affecting developing countries like Ethiopia. To initiate effective prevention and control of CE, comprehensive data on the local lifecycles of the various species/genotypes of Echinococcus are needed. In the present study, conducted in eastern Ethiopia, a total of 1106 livestock animals were examined at three slaughterhouses, which resulted in combined prevalence of morphologically and molecularly confirmed CE of 8.4% (75/891) in cattle, 1.1% (1/95) in sheep, 0.0% (0/95) in goats and 12.0% (3/25) in camels. All cystic lesions recovered during post mortem examination were assessed for cyst condition and underwent molecular characterization by PCR and sequencing of a 1081â¯bp fragment of the mitochondrial cox1 gene. A total of 175 cysts belonged to E. granulosus s.s. (nâ¯=â¯165), E. ortleppi (nâ¯=â¯6) and E. canadensis G6/7 (nâ¯=â¯4). Of all examined cysts, only 14 were fertile and contained protoscoleces, all from the lungs of cattle: 5 were E. granulosus s.s., 6 E. ortleppi and 3 E. canadensis G6/7. In sheep, only one sterile liver cyst of E. granulosus s.s. was found, while in camels seven sterile or caseated/calcified cysts of E. granulosus s.s. and E. canadensis G6/7 were found in liver and lungs. In conclusion, the prevalence of CE was rather low compared to other regions of Ethiopia, and, based on the number of fertile cysts, three Echinococcus spp. contributed almost equally to transmission. Cattle seem to be, epidemiologically, the most important livestock species. Our data provide a substantial basis for more detailed investigations of the transmission dynamics of CE in the study area.
Assuntos
Equinococose/veterinária , Echinococcus/genética , Gado/parasitologia , Matadouros , Animais , Camelus/parasitologia , Bovinos , Doenças dos Bovinos/epidemiologia , Doenças dos Bovinos/parasitologia , Equinococose/epidemiologia , Equinococose/parasitologia , Echinococcus/classificação , Complexo IV da Cadeia de Transporte de Elétrons/genética , Etiópia/epidemiologia , Genes Mitocondriais , Doenças das Cabras/epidemiologia , Doenças das Cabras/parasitologia , Cabras , Doenças Negligenciadas/epidemiologia , Doenças Negligenciadas/parasitologia , Doenças Negligenciadas/veterinária , Prevalência , Ovinos , Doenças dos Ovinos/epidemiologia , Doenças dos Ovinos/parasitologia , Zoonoses/epidemiologia , Zoonoses/parasitologiaRESUMO
Cystic Echinococcosis (CE) is a widespread neglected zoonotic disease and is caused by the larval stage of the dog tapeworm Echinococcus granulosus sensu lato. CE is more frequent in livestock-rearing areas and where people live a nomadic or seminomadic lifestyle such as in Kajiado County, Kenya. There is limited data on CE disease situation in the county of Maasailand; the present study, therefore, reports on the prevalence of CE in cattle, sheep, and goats and their relative importance in CE transmission in Kajiado County. In total, 1,486 livestock (388 cattle, 625 sheep, and 473 goats) slaughtered in two abattoirs were examined for the presence of hydatid cysts in various organs. Cyst isolates were genotyped by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) of the NADH dehydrogenase subunit 1 gene (nad1). The overall prevalence of CE was 14.8% (220/1486), while prevalence per livestock species was 15.2% (72/473) in goats, 14.9% (93/625) in sheep, and 14.2% (55/388) in cattle. Out of the 421 cysts isolated, 389 cysts were successfully characterized to be either E. granulosus sensu stricto (s. s.), 356/389 (91.5%), E. canadensis (G6/7), 26/389 (6.7%), or E. ortleppi, 7/389 (1.8%). This record confirms predominance of E. granulosus s. s. in Maasailand and other parts of Kenya, while the importance of E. ortleppi and E. canadensis (G6/7) to the general CE burden in Maasailand might be higher than previously thought. More so, a higher infection pressure for humans by E. granulosus s. s. based on its abundance could be speculated. The study sheds significant light on CE situation in livestock in the nomadic/seminomadic society of the Maasai in Kajiado County and provides good bases to investigate human CE in the area.
Assuntos
Equinococose , Echinococcus , Técnicas de Genotipagem , Proteínas de Helminto/genética , Helmintíase Animal , Gado/parasitologia , Animais , Bovinos , Cães , Equinococose/diagnóstico , Equinococose/epidemiologia , Equinococose/genética , Echinococcus/classificação , Echinococcus/genética , Cabras , Helmintíase Animal/diagnóstico , Helmintíase Animal/epidemiologia , Helmintíase Animal/genética , Humanos , Quênia/epidemiologia , Polimorfismo de Fragmento de Restrição , OvinosRESUMO
We report a sheep infected with Echinococcus canadensis G8 tapeworm in China in 2018. This pathogen was previously detected in moose, elk, muskox, and mule deer in Europe and North America; our findings suggest a wider host range and geographic distribution. Surveillance for the G8 tapeworm should be conducted in China.
Assuntos
Equinococose/veterinária , Echinococcus , Doenças dos Ovinos/epidemiologia , Doenças dos Ovinos/parasitologia , Animais , China/epidemiologia , Echinococcus/classificação , Echinococcus/genética , Echinococcus/isolamento & purificação , Genes Mitocondriais , Genótipo , História do Século XXI , Humanos , Filogenia , Ovinos , Doenças dos Ovinos/históriaRESUMO
This study examined Echinococcus spp. genotypes and genetic variants isolated from humans as well as domestic and wild animals from the Qinghai-Tibetan Plateau Area using the cox1 gene. All samples except the pika isolates were identified as the Echinococcus granulosus sensu stricto. Sixteen different haplotypes with considerable intraspecific variation were detected and characterized in mitochondrial cox1 sequences. The parsimonious network of cox1 haplotypes showed star-like features, and the neutrality indexes computed via Tajima's D and Fu's Fs tests showed high negative values in E. granulosus s. s., indicating deviations from neutrality; the Fst values were low among the populations, implying that the populations were not genetically differentiated. The pika isolates were identified as E. multilocularis and E. shiquicus. Only one haplotype was recognized in the pika isolates. E. granulosus s. s. was the predominant species found in animals and humans, followed by E. multilocularis and E. shiquicus, with high genetic diversity circulating among the animals and humans in this area. Further studies are needed to cover many sample collection sites and larger numbers of pathogen isolates, which may reveal abundant strains and/or other haplotypes in the hydatid cysts infecting human and animal populations of the QTPA, China.
Assuntos
Equinococose/parasitologia , Equinococose/veterinária , Echinococcus/classificação , Echinococcus/genética , Variação Genética , Genótipo , Animais , Animais Domésticos , Animais Selvagens , China , Echinococcus/isolamento & purificação , Complexo IV da Cadeia de Transporte de Elétrons/genética , Haplótipos , HumanosRESUMO
Larval tapeworms of Echinococcus granulosus have been viewed as the etiological agent for the zoonotic disease cystic echinococcosis, but the species is a complex readily divided into several species and genotypes. Cystic echinococcosis is an important public health issue. Here, the case of liver hydatid cyst in a donkey and molecular characterization of the cyst is presented. The fluid-filled hydatid cyst materials were obtained from the liver of a necropsied donkey. Genomic DNA was extracted and PCR amplification of mitochondrial 12S rRNA gene as well as partial sequencing of mitochondrial cytochrome c oxidase subunit 1 (COI) gene were performed. All cysts were fertile. Traditional polymerase chain reaction (PCR) amplification of 12S rRNA and COI yielded bands (254 and 446 base pairs, respectively) for all 3 cyst samples. However, partial COI gene sequences were identical to those reported for Echinococcus equinus (formerly E. granulosus genotype G4). Thus E. equinus is still transmitting among the equids in Turkey but the mitochondrial 12S rRNA gene primers may not be sufficient for the molecular characterization of members of the E. granulosus species/genotype complex. Molecular diagnosis must be confirmed by partial COI sequence analysis.