RESUMEN
Typical Kunitz proteins (I2 family of the MEROPS database, Kunitz-A family) are metazoan competitive inhibitors of serine peptidases that form tight complexes of 1:1 stoichiometry, mimicking substrates. The cestode Echinococcus granulosus, the dog tapeworm causing cystic echinococcosis in humans and livestock, encodes an expanded family of monodomain Kunitz proteins, some of which are secreted to the dog host interface. The Kunitz protein EgKU-7 contains, in addition to the Kunitz domain with the anti-peptidase loop comprising a critical arginine, a C-terminal extension of â¼20 amino acids. Kinetic, electrophoretic, and mass spectrometry studies using EgKU-7, a C-terminally truncated variant, and a mutant in which the critical arginine was substituted by alanine, show that EgKU-7 is a tight inhibitor of bovine and canine trypsins with the unusual property of possessing two instead of one site of interaction with the peptidases. One site resides in the anti-peptidase loop and is partially hydrolyzed by bovine but not canine trypsins, suggesting specificity for the target enzymes. The other site is located in the C-terminal extension. This extension can be hydrolyzed in a particular arginine by cationic bovine and canine trypsins but not by anionic canine trypsin. This is the first time to our knowledge that a monodomain Kunitz-A protein is reported to have two interaction sites with its target. Considering that putative orthologs of EgKU-7 are present in other cestodes, our finding unveils a novel piece in the repertoire of peptidase-inhibitor interactions and adds new notes to the evolutionary host-parasite concerto.
Asunto(s)
Echinococcus granulosus , Proteínas del Helminto , Echinococcus granulosus/enzimología , Echinococcus granulosus/genética , Echinococcus granulosus/metabolismo , Animales , Perros , Proteínas del Helminto/metabolismo , Proteínas del Helminto/genética , Proteínas del Helminto/química , Inhibidores de Tripsina/metabolismo , Inhibidores de Tripsina/química , Bovinos , Secuencia de Aminoácidos , Tripsina/química , Tripsina/metabolismoRESUMEN
Cestodes use own lipid-binding proteins to capture and transport hydrophobic ligands, including lipids that they cannot synthesise as fatty acids and cholesterol. In E. granulosus s.l., one of these lipoproteins is antigen B (EgAgB), codified by a multigenic and polymorphic family that gives rise to five gene products (EgAgB8/1-5 subunits) assembled as a 230 kDa macromolecule. EgAgB has a diagnostic value for cystic echinococcosis, but its putative role in the immunobiology of this infection is still poorly understood. Accumulating research suggests that EgAgB has immunomodulatory properties, but previous studies employed denatured antigen preparations that might exert different effects than the native form, thereby limiting data interpretation. This work analysed the modulatory actions on macrophages of native EgAgB (nEgAgB) and the recombinant form of EgAg8/1, which is the most abundant subunit in the larva and was expressed in insect S2 cells (rEgAgB8/1). Both EgAgB preparations were purified to homogeneity by immunoaffinity chromatography using a novel nanobody anti-EgAgB8/1. nEgAgB and rEgAgB8/1 exhibited differences in size and lipid composition. The rEgAgB8/1 generates mildly larger lipoproteins with a less diverse lipid composition than nEgAgB. Assays using human and murine macrophages showed that both nEgAgB and rEgAgB8/1 interfered with in vitro LPS-driven macrophage activation, decreasing cytokine (IL-1ß, IL-6, IL-12p40, IFN-ß) secretion and ·NO generation. Furthermore, nEgAgB and rEgAgB8/1 modulated in vivo LPS-induced cytokine production (IL-6, IL-10) and activation of large (measured as MHC-II level) and small (measured as CD86 and CD40 levels) macrophages in the peritoneum, although rEgAgB8/1 effects were less robust. Overall, this work reinforced the notion that EgAgB is an immunomodulatory component of E. granulosus s.l. Although nEgAgB lipid's effects cannot be ruled out, our data suggest that the EgAgB8/1 subunit contributes to EgAgB´s ability to regulate the inflammatory activation of macrophages.
Asunto(s)
Echinococcus granulosus , Humanos , Animales , Ratones , Echinococcus granulosus/genética , Echinococcus granulosus/metabolismo , Interleucina-6/metabolismo , Lipopolisacáridos/metabolismo , Activación de Macrófagos , Lipoproteínas/genética , Lipoproteínas/metabolismo , Macrófagos , Citocinas/metabolismoRESUMEN
Cystic echinococcosis (CE) is a zoonotic disease caused by the tapeworm Echinococcus granulosus sensu lato (s.l). In the intermediate host, this disease is characterized by the growth of cysts in viscera such as liver and lungs, inside of which the parasite develops to the next infective stage known as protoscoleces. There are records that the infected viscera affect the development and morphology of E. granulosus s.l. protoscolex in hosts such as buffalo or humans. However, the molecular mechanisms that drive these differences remains unknown. Weighted gene co-expression network analysis (WGCNA) using a set of RNAseq data obtained from E. granulosus sensu stricto (s.s.) protoscoleces found in liver and lung cysts reveals 34 modules in protoscoleces of liver origin, of which 12 have differential co-expression from protoscoleces of lung origin. Three of these twelve modules contain hub genes related to immune evasion: tegument antigen, tegumental protein, ubiquitin hydrolase isozyme L3, COP9 signalosome complex subunit 3, tetraspanin CD9 antigen, and the methyl-CpG-binding protein Mbd2. Also, two of the twelve modules contain only hypothetical proteins with unknown orthology, which means that there are a group of unknown function proteins co-expressed inside the protoscolex of liver CE cyst origin. This is the first evidence of gene expression differences in protoscoleces from CE cysts found in different viscera, with co-expression networks that are exclusive to protoscoleces from liver CE cyst samples. This should be considered in the control strategies of CE, as intermediate hosts can harbor CE cysts in liver, lungs, or both organs simultaneously.
Asunto(s)
Quistes , Equinococosis , Echinococcus granulosus , Humanos , Animales , Echinococcus granulosus/genética , Evasión Inmune , Genotipo , Equinococosis/genética , Equinococosis/parasitologíaRESUMEN
Nuclear receptors (NRs) are ligand-modulated transcription factors that regulate various biological processes, such as metabolism, development and reproduction. Although NRs with two DNA-binding domains (2DBD) were identified in Schistosoma mansoni (Platyhelminth, Trematoda) more than fifteen years ago, these proteins have been poorly studied. 2DBD-NRs could become attractive therapeutic targets to combat parasitic diseases such as cystic echinococcosis since this type of protein is absent in vertebrate hosts. Cystic echinococcosis is a worldwide zoonosis caused by the larval stage of the parasitic platyhelminth Echinococcus granulosus (Cestoda) that generates an important public health problem and a significant economic loss. Recently, our research group identified four 2DBD-NRs in E. granulosus, named Eg2DBDα, Eg2DBDα.1 (an isoform of Eg2DBDα), Eg2DBDß, and Eg2DBDγ. This work demonstrated that Eg2DBDα.1 forms homodimers through the E and F regions, whereas its interaction with EgRXRßa could not be detected. In addition, the stimulation of Eg2DBDα.1 homodimerization by intermediate host serum was shown, suggesting that at least one lipophilic molecule from bovine serum could bind to Eg2DBDα.1. Finally, Eg2DBDs expression studies in the protoscolex larval stage were performed, indicating that Eg2dbdγ is not expressed, whereas Eg2dbdα has the highest expression level followed by Eg2dbdß and Eg2dbdα.1 in decreased order. Overall, these findings provide new insights into the mechanism of action of Eg2DBDα.1 and its potential role in host-parasite communication.
Asunto(s)
Equinococosis , Echinococcus granulosus , Parásitos , Animales , Echinococcus granulosus/genética , Parásitos/metabolismo , Dimerización , Equinococosis/parasitología , Larva/genética , Larva/metabolismo , Receptores Citoplasmáticos y Nucleares/genética , Receptores Citoplasmáticos y Nucleares/metabolismoRESUMEN
Most human cystic echinococcosis (CE) cases worldwide are attributed to Echinococcus granulosus sensu stricto (s.s), followed by the G6 and G7 genotypes. While E. granulosus s.s. has a cosmopolitan distribution, the G6 genotype is restricted to areas where camels and goats are present. Goats are the primary livestock in the Neuquén province in Argentina where the G6 genotype has been reported to be responsible for a significant percentage of CE human cysts genotyped. In the present study, we genotyped 124 Echinococcus cysts infecting 90 CE-confirmed patients. Echinococcus granulosus s.s. was identified in 51 patients (56.7%) with 81 cysts and the G6 genotype in 39 patients (43.3%) harbouring 43 cysts. Most CE cases ≤18 years were male suggesting pastoral work could be a risk factor for the infection. Echinococcus granulosus s.s. was significantly found more frequently in the liver (32/51 patients) and the G6 genotype in the lungs and extrahepatic localizations (27/39). The patients infected with E. granulosus s.s., presented up to 6 cysts while patients infected with G6 presented a maximum of 2. The diameter of lung cysts attributed to E. granulosus s.s. was significantly larger compared to lung cysts from G6. Following the WHO ultrasound classification of liver cysts, we observed inactive cysts in 55.6% of G6 cysts and only 15.3% of E. granulosus s.s cysts. In conclusion, we provide evidence of differences in clinical aspects of CE caused by E. granulosus s.s. and the G6 genotype of E. granulosus s.l. complex infecting humans.
Asunto(s)
Quistes , Equinococosis , Echinococcus granulosus , Animales , Humanos , Masculino , Femenino , Echinococcus granulosus/genética , Argentina/epidemiología , Equinococosis/epidemiología , Equinococosis/veterinaria , Genotipo , Cabras , CamelusRESUMEN
Glutathione transferase enzymes (GSTs) are believed to be a major detoxification system in helminth parasites and have been associated with immunomodulation of the host response. Echinococcus granulosus sensu lato (s.l.) is a cestode parasite known to express at least five different GSTs, but no Omega-class enzymes have been reported in this parasite or in any other cestode. Herein we report the identification of a new member of the GST superfamily in E. granulosus s.l., which is phylogenetically related to the Omega-class: EgrGSTO. Through mass spectrometry, we showed that the 237 amino acids protein EgrGSTO is expressed by the parasite. Moreover, we identified homologues of EgrGSTO in other eight members of the Taeniidae family, including E. canadensis, E. multilocularis, E. oligarthrus, Hydatigera taeniaeformis, Taenia asiatica, T. multiceps, T. saginata and T. solium. A manual sequence inspection and rational modification yielded eight Taeniidae's GSTO sequences, each one encoding for a 237 aa polypeptide showing 80.2% overall identity. To the best of our knowledge, this is the first description of genes encoding for Omega-class GSTs in worms belonging to the Taeniidae family -that at least in E. granulosus s.l. is expressed as a protein- suggesting the gene encodes for a functional protein.
Asunto(s)
Echinococcus granulosus , Parásitos , Taenia , Animales , Glutatión Transferasa/química , Echinococcus granulosus/genética , Parásitos/metabolismoRESUMEN
Cystic echinococcosis (CE) is a zoonosis caused by species of the complex Echinococcus granulosus, sensu lato in their larval stage. It is an endemic disease in the province of Río Negro, where small farmers generally have both sheep and goats. Lamb vaccination with EG95 was incorporated in 2009 with very good results: in fact, it contributed to a significant drop in prevalence of infection in both sheep and goats, when determined by necropsy and serology in 2018. In the design of the activity, it was decided not to vaccinate goats in order to minimize the operational requirements of vaccination and comments from producers about the rarity of observing hydatid cysts in goat viscera were considered. OBJECTIVE: To identify causes which can still generate infection in dogs, and to detect species/genotypes in circulation in the province of Río Negro. MATERIALS AND METHODS: In indigenous reserves comprised within the area of lamb vaccination with 3 doses of EG95, (dose 1 in December, dose 2 in January and dose 3 in December of the year following, at the time of application of dose 1 to the new lambs). Prevalence in adult goats and sheep was determined by necropsy and serology (ELISA). Infective species/genotypes present in the work area and in the rest of the province of Río Negro were identified by Cox1 mitochondrial gene sequencing. Epidemiological analysis was completed with surveys among farmers about slaughter habits for human consumption. RESULTS: Through serology and necropsy, infection rates in vaccinated and nonvaccinated sheep were significantly different (21% versus 66%). Non-vaccinated sheep and non-vaccinated goats were also significantly different in that there was less infection in goats compared to sheep (7% versus 66% for necropsy, 30% versus 61% for serology); After many years of sheep vaccination the infection positives were low, and differences between vaccinated sheep and non-vaccinated goats turned out non-significant (21% versus 7%). With reference to epidemiology and control along the period 2018-2022, PZQ dosing of dogs 4 times a year was maintained, and 2 extra deworming tasks were introduced together with dose 1 and 2 of EG95, performed by the veterinary vaccination team, ensuring the ingestion of PZQ by dogs. Assessment of animal slaughter for consumption in 41 producers showed that 21 of them slaughter a monthly average of 18 goats (an average of 0.43 goat per month per farm) and 36 in all slaughter 35 old sheep in a year (average of 0.85 sheep per month per farm). With respect to identification of species/genotypes as from 2010, genotypes G1 have been found in 11 sheep (out of which 6 belong to vaccination zone) and genotypes G7, in one pig. A goat cyst within vaccination zone turned out unfertile and it was not possible to sequence it. CONCLUSION: Design and implementation of a vaccine programme combined with the use of PZQ resulted as cost-effective, since it was possible to maintain the vaccine over time, with clear impact on prevalence decrease in sheep and goats.
Asunto(s)
Enfermedades de los Perros , Equinococosis , Echinococcus granulosus , Enfermedades de las Cabras , Enfermedades de las Ovejas , Enfermedades de los Porcinos , Vacunas , Negro o Afroamericano , Animales , Argentina/epidemiología , Enfermedades de los Perros/epidemiología , Enfermedades de los Perros/prevención & control , Perros , Equinococosis/epidemiología , Equinococosis/prevención & control , Equinococosis/veterinaria , Echinococcus granulosus/genética , Enfermedades de las Cabras/epidemiología , Enfermedades de las Cabras/prevención & control , Cabras , Humanos , Ovinos , Enfermedades de las Ovejas/epidemiología , Enfermedades de las Ovejas/prevención & control , Porcinos , Vacunación/veterinariaRESUMEN
BACKGROUND: Knowledge of the distribution cystic is required for its territorial control. Aim: To describe the spatial distribution of Echinococcus granulosus sensu lato genotypes by host in the American continent. MATERRIAL AND METHODS: A systematic review of studies from the American continent, related to genotypes of the E. granulosus s.l complex were included, including any host species, without restriction of language or year of publication. Sensitive searches were performed based on sensitive searches from PubMed, EMBASE, ScienceDirect, SCOPUS and WoS; SciELO and BIREME-BVS and Trip Database. MeSH and free terms were used, including articles up to December 2020. Cartography was carried out with the Arc Map 10® program, using a world geodetic system. Result variables sought were genotype, host, geographic location, year of publication, number of samples, genes used for genotyping. RESULTS: From 1123 retrieved studies retrieved, 53 met the inclusion and exclusion criteria. The studies analyzed represent 3,397 samples from humans and animals. Thirty six percent of articles were published in the five-year period 2016-2020. Reports were mainly from Argentina (27.9%), Brazil (20.6%) and Chile (13.2%). The most reported genotypes globally were G1-G3 (47.3%), G7 (15.3%), G5 (14.6%) and G6 (13.3%). A predominance of G1-G3 and G6 genotypes was verified in South America, G8 and G10 in North America, and "epidemiological silence" in Central America and the Caribbean. Conclusions: Spatial analysis allows defining the relationship of territories and cases with their own characteristics, which can help to plan control interventions.
Asunto(s)
Humanos , Echinococcus granulosus/genética , Equinococosis , Argentina/epidemiología , Brasil , Genotipo , AnimalesRESUMEN
Cystic echinococcosis (CE) is a zoonotic disease worldwide distributed, caused by the cestode Echinococcus granulosus sensu lato (E. granulosus), with an incidence rate of 50/100,000 person/year and a high prevalence in humans of 5-10%. Serology has variable sensitivity and specificity and low predictive values. Antigens used are from the hydatid fluid and recombinant antigens have not demonstrated superiority over hydatid fluid. A cell line called EGPE was obtained from E. granulosus sensu lato G1 strain from bovine liver. Serum from CE patients recognizes protein extracts from EGPE cells with higher sensitivity than protein extracts from hydatid fluid. In the present study, EGPE cell protein extracts and supernatants from cell colonies were eluted from a protein G affinity column performed with sera from 11 CE patients. LC-MS/MS proteomic analysis of the eluted proteins identified four E. granulosus histones: one histone H4 in the cell extract and supernatant, one histone H2A only in the cell extract, and two histones H2A only in the supernatant. This differential distribution of histones could reflect different parasite viability stages regarding their role in gene transcription and silencing and could interact with host cells. Bioinformatics tools characterized the linear and conformational epitopes involved in antibody recognition. The three-dimensional structure of each histone was obtained by molecular modeling and validated by molecular dynamics simulation and PCR confirmed the presence of the epitopes in the parasite genome. The three histones H2A were very different and had a less conserved sequence than the histone H4. Comparison of the histones of E. granulosus with those of other organisms showed exclusive regions for E. granulosus. Since histones play a role in the host-parasite relationship they could be good candidates to improve the predictive value of serology in CE.
Asunto(s)
Quistes , Equinococosis , Echinococcus granulosus , Animales , Bovinos , Extractos Celulares , Cromatografía Liquida , Equinococosis/epidemiología , Equinococosis/parasitología , Echinococcus granulosus/genética , Epítopos de Linfocito B , Genotipo , Histonas , Humanos , Hígado , Hepatopatías , Proteómica , Espectrometría de Masas en TándemRESUMEN
Glutathione transferases (GSTs) perform catalytic and non-catalytic activities, mostly involved in stress-response and cell detoxification. Helminth parasites express several GSTs of multiple classes that are involved in the neutralization of potentially harmful oxidants, and in the inactivation or removal of xenobiotics. Additionally, GSTs participate in immunomodulatory processes that facilitate the parasite establishment and survival within its host. In Echinococcus granulosus sensu lato (s.l.) -the cestode parasite responsible for cystic echinococcosis- only one Mu-class GST has been reported. In the present work, by using bioinformatic and proteomic approaches we searched for novel Mu-class GSTs potentially involved in the parasite oxidative-stress metabolism. In the genome of E. granulosus s.l., 6 GST-related sequences were found to constitute a strongly conserved phylogenetical clade with Mu-class members. Among them, 5 displayed conserved gene structure (exon/intron), as well as specific residues and motifs characteristic of Mu-class enzymes. By proteomic analysis, 3 Mu-GSTs were identified to be expressed in the protoscolex parasite stage, 2 of them being firstly described as Mu-class GSTs here. The existence of more than one productive Mu-GST gene expands the parasite xenobiotic phase II metabolism, which might have beneficial roles on E. granulosus s.l. ability to successfully infect its host.
Asunto(s)
Equinococosis , Echinococcus granulosus , Parásitos , Animales , Equinococosis/parasitología , Echinococcus granulosus/genética , Glutatión Transferasa/genética , Glutatión Transferasa/metabolismo , Parásitos/metabolismo , ProteómicaRESUMEN
Cystic Echinococcosis (CE), a zoonotic parasitic disease, is caused by the cestode Echinococcus granulosus sensu lato. CE inflicts severe damage in cattle, sheep, and human hosts worldwide. Fertile CE cysts are characterized by the presence of viable protoscoleces. These parasite forms are studied with minimal contamination with host molecules. Hosts, cattle and sheep, show differences in their CE cyst fertility. The effect of the host in protoscolex transcriptome is not known. We genotyped and performed transcriptomic analysis on sheep protoscoleces obtained from liver and lung CE cysts. The transcriptomic data of Echinococcus granulosus sensu stricto protoscoleces from 6 lung CE cysts and 6 liver CE cysts were Collected. For host comparison analysis, 4 raw data files belonging to Echinococcus granulosus sensu stricto protoscoleces from cattle liver CE cysts were obtained from the NCBI SRA database. Principal component and differential expression analysis did not reveal any statistical differences between protoscoleces obtained from liver or lung cysts, either within the same sheep or different sheep hosts. Conversely, there are significant differences between cattle and sheep protoscolex samples. We found differential expression of immune-related genes. In cattle, 7 genes were upregulated in protoscoleces from liver cysts. In sheep, 3 genes were upregulated in protoscoleces from liver and lung CE cysts. Noteworthy, are the differential expression of antigen B, tegument antigen, and arginase-2 in samples obtained from sheep CE cysts, and basigin in samples from cattle CE cysts. These findings suggest that the host species is an important factor involved in the differential expression of immune related genes, which in turn is possibly related to the fertility of Echinococcus granulosus sensu stricto cysts.
Asunto(s)
Enfermedades de los Bovinos , Quistes , Equinococosis , Echinococcus granulosus , Enfermedades de las Ovejas , Animales , Bovinos , Enfermedades de los Bovinos/parasitología , Quistes/veterinaria , Equinococosis/parasitología , Equinococosis/veterinaria , Echinococcus granulosus/genética , Expresión Génica , Perfilación de la Expresión Génica/veterinaria , Genotipo , Ovinos , Enfermedades de las Ovejas/genética , Enfermedades de las Ovejas/parasitologíaRESUMEN
INTRODUCTION: The aim of this study was to develop a synthesis of the evidence available regarding verified E. granulosus sensu lato (s.l.) genotypes in different species worldwide. MATERIAL AND METHODS: A systematic review was performed including studies concerning genotypes of E. granulosus s.l. without language or genotyped method restriction, published between 1990 and 2020. A systematic search was carried out in Trip Database, BIREME, SciELO, LILACS, IBECS, PAHO-WHO, EMBASE, PubMed, Scopus, and WoS. Variables of interest were year of publication, country, number of samples, and hosts; genotypes, molecular marker, haplotypes and molecular biology techniques used. Descriptive statistics were applied. RESULTS: 2411 articles were analyzed, however 135 met the selection criteria, representing 8643 liver and lung samples. Of the samples selected 24% were human, the remaining samples pertained to non-human animal hosts; cattle and sheep prevailed with 28.6% and 26.6% of the studied samples, respectively. The reported evidence is mainly from Iran, Turkey, Argentina, China and Chile; with 50, 11, 6, 6 and 5 studies, respectively, published between 1992 and 2020 [most frequently during 2015-2020 (76/135 studies; 56.3%)]. The mitochondrial gene cox1 was generally sequenced and informative (91.8%). Genotypes most frequently identified were E. granulosus sensu stricto (s.s.) (83.2%). CONCLUSIONS: Based on this overall evidence, it can be concluded that publications related to genotypes of E. granulosus s.l. are heterogeneous. E. granulosus ss accounts for the vast majority of the global burden of E. granulosus s.l. worldwide. Further studies including larger number of cases and adequate internal validity are required to specify the distribution of genotypes in various host species. TRIAL REGISTRATION: PROSPERO CRD42018099827.
Asunto(s)
Equinococosis , Echinococcus granulosus , Animales , Bovinos , Equinococosis/parasitología , Equinococosis/veterinaria , Echinococcus granulosus/genética , Genes Mitocondriales , Genotipo , Humanos , OvinosRESUMEN
BACKGROUND: Knowledge of the distribution cystic is required for its territorial control. AIM: To describe the spatial distribution of Echinococcus granulosus sensu lato genotypes by host in the American continent. MATERIAL AND METHODS: A systematic review of studies from the American continent, related to genotypes of the E. granulosus s.l complex were included, including any host species, without restriction of language or year of publication. Sensitive searches were performed based on sensitive searches from PubMed, EMBASE, ScienceDirect, SCOPUS and WoS; SciELO and BIREME-BVS and Trip Database. MeSH and free terms were used, including articles up to December 2020. Cartography was carried out with the Arc Map 10® program, using a world geodetic system. Result variables sought were genotype, host, geographic location, year of publication, number of samples, genes used for genotyping. RESULTS: From 1123 retrieved studies retrieved, 53 met the inclusion and exclusion criteria. The studies analyzed represent 3,397 samples from humans and animals. Thirty six percent of articles were published in the five-year period 2016-2020. Reports were mainly from Argentina (27.9%), Brazil (20.6%) and Chile (13.2%). The most reported genotypes globally were G1-G3 (47.3%), G7 (15.3%), G5 (14.6%) and G6 (13.3%). A predominance of G1-G3 and G6 genotypes was verified in South America, G8 and G10 in North America, and "epidemiological silence" in Central America and the Caribbean. CONCLUSIONS: Spatial analysis allows defining the relationship of territories and cases with their own characteristics, which can help to plan control interventions.
Asunto(s)
Equinococosis , Echinococcus granulosus , Animales , Humanos , Echinococcus granulosus/genética , Argentina/epidemiología , Brasil , GenotipoRESUMEN
The prevalence of cystic echinococcosis is high in many livestock areas of Peru, where intermediate hosts such as sheep, cattle, and South American camelids can be infected. Several species of E. granulosus have been described in relation to its genetic diversity and distribution. The aim of this study was to determine the species of E. granulosus sensu lato (s.l.) metacestodes collected from sheep, cattle, swine and camelids at different localities in the department of Puno, in the southern highlands of Peru. One hundred and fifty-two echinococcal cysts were collected from 10 different locations. E. granulosus s.l. species were determined by amplification of the Internal transcribed spacer 1 of the ribosomal DNA using a Nested PCR-RFLP technique. The cytochrome C oxidase 1 gene (450 bp) was also amplified and sequenced in samples with different RFLP patterns. Cysts samples were collected from sheep (39.5%), cattle (32.9%), pigs (15.8%) and alpacas/llamas (11.8%). E. granulosus sensu stricto (G1 genotype) was mainly identified in all animal hosts, while, the E. canadensis (G7) was only identified in cysts from pigs and alpacas. This is the first report of E. granulosus sensu stricto and E. canadensis in llamas and alpacas, respectively. Knowledge of species and molecular epidemiology of E. granulosus s.l. in endemic areas in Peru may help to evaluate preventive programs, understand disease transmission, as well as improve vaccine and chemotherapy effectiveness.
Asunto(s)
Equinococosis , Echinococcus granulosus , Echinococcus , Animales , Bovinos , Equinococosis/epidemiología , Equinococosis/veterinaria , Echinococcus granulosus/genética , Genotipo , Ganado , Perú/epidemiología , Ovinos , PorcinosRESUMEN
Viable eggs of the canine intestinal tapeworm Echinococcus granulosus sensu lato (s.l.) infect various intermediate hosts causing cystic echinococcosis (CE). Furthermore, CE represents a serious zoonosis causing a significant global burden of disease. CE is highly endemic in South America, including Argentina, Brazil, Chile, Uruguay, and Peru. For Bolivia, no official data concerning the incidence in humans or the number of livestock and dogs infected are available. However, it is well known that CE occurs in Bolivia. We aim here to fill the gap in the current knowledge of the epidemiological situation of CE in Bolivia, providing a historical overview of documents published within the country, which have never been comprehensively reviewed. The very first documentation of E. granulosus infection in animals dates in 1910, while the first human case was reported in 1913. In total, 876 human CE cases have been reported in the scientific literature, with an apparent increase since the 1970s. In the absence of other epidemiological studies, the highest prevalence in human comes from Tupiza, Potosí Department, where 4.1% (51/1,268) of the population showed signs of CE at mass ultrasound screening in 2011. In the same report, 24% of dog faecal samples were positive for coproantigens of E. granulosus s.l. in ELISA. The highest prevalence in intermediate hosts reported at abattoir reached 37.5% in cattle from Potosí, followed by 26.9% in llamas from Oruro, 2.4% in pigs and 1.4% in sheep from La Paz. Finally, Echinococcus granulosus sensu stricto (s.s.), Echinococcus ortleppi (G5), and Echinococcus intermedius (G7) have been identified in Bolivia. Data reviewed here confirm that E. granulosus s.l. is circulating in Bolivia and that a proper prospective nationwide epidemiological study of CE is urgently needed to define transmission patterns as a basis for the planning and implementation of future control measurements.
Asunto(s)
Equinococosis/veterinaria , Echinococcus granulosus/genética , Zoonosis/parasitología , Animales , Bolivia/epidemiología , Equinococosis/epidemiología , Humanos , Vigilancia de la Población , Zoonosis/epidemiologíaRESUMEN
Glutathione transferases (GSTs) belong to a diverse superfamily of multifunctional proteins involved in metabolic detoxification. In helminth parasite, GSTs are particularly relevant since they are also involved in host immunomodulation. Echinococcus granulosus sensu lato (s.l.) is a cestode parasite known to express at least three phylogenetically distant cytosolic GSTs: EgGST1 and EgGST2 previously grouped within Mu and Sigma classes, respectively; and EgGST3 related to both Omega and Sigma classes. To better characterize E. granulosus s.l. GSTs, herein their expression and distribution were assessed in the pre-adult protoscolex (PSC) parasite stage. Potential transcriptional regulatory mechanisms of the corresponding EgGST genes were also explored. Firstly, the transcription of the three EgGSTs was significantly induced during the early stages of the murine model of infection, suggesting a potential role during parasite establishment. EgGST1 was detected in the parenchyma of PSCs and its expression increased after H2O2 exposure, supporting its role in detoxification. EgGST2 was mainly detected on the PSCs tegument, strategically localized for potential immunoregulation functions due to its Sigma-class characteristics. In addition, its expression increased after anthelmintic treatment, suggesting a role in chemotherapy resistance. Finally, the Omega-related EgGST3 was localized throughout the entire PSC body, including suckers and tegument, and since its expression also increased after H2O2 treatment, a potential role in oxidative stress response could also be ascribed. On the other hand, known cis-acting regulatory motifs were detected in EgGST genes, suggesting similar transcription processes to other eukaryotes. The results herein reported provide additional data regarding the roles of EgGSTs in E. granulosus s.l. biology, contributing to a better understanding of its host-parasite interaction.
Asunto(s)
Echinococcus granulosus , Animales , Antihelmínticos , Echinococcus granulosus/genética , Echinococcus granulosus/metabolismo , Glutatión Transferasa/genética , Glutatión Transferasa/metabolismo , Peróxido de Hidrógeno , Ratones , Estrés OxidativoRESUMEN
Resumen Introducción: La evidencia sobre las características genotípicas de la infección por Echinococcus granulosus en humanos es escasa. Objetivo: Desarrollar un resumen de la evidencia disponible respecto a genotipos de E. granulosus verificados en hidatidosis humana en el mundo. Material y Métodos: Revisión sistemática. Se incluyeron artículos relacionados con genotipos de E. granulosus, en humanos, sin restricción de lenguaje ni método de secuenciación; publicados entre 1990-2019. Se realizó una búsqueda sistemática en WoS, EMBASE, MEDLINE, SCOPUS, Trip Database, BIREME, SciELO, LILACS, IBECS y OPS-OMS. Las variables en estudio fueron: año de publicación, país de origen, número de muestras, órganos parasitados, marcador molecular utilizado y genotipo identificado. Se aplicó estadística descriptiva. Resultados: Se identificaron 701 artículos relacionados; 62 cumplieron los criterios de selección, representando 1.511 muestras. La evidencia existente fue publicada entre 1994 y 2019 y proviene principalmente de Irán (45,2%). El método de secuenciación más utilizado fue amplificación por reacción de polimerasa en cadena más secuenciación tipo Sanger con genotipificación del gen cox1 (79,0%). Los genotipos identificados con mayor frecuencia fueron G1 (49,1%) y el complejo G1/G3 (32,2%). Conclusión: Las publicaciones relacionadas con genotipos de E. granulosus en humanos son escasas y heterogéneas. Eg G1 representa la mayor parte de la carga global mundial.
Abstract Background: The evidence regarding genotypic characteristics of Echinococcus granulosus infection in humans worldwide is scarce. Aim: To develop a synthesis of the available evidence regarding genotypes of E. granulosus verified in humans worldwide. Methods: Systematic review. Articles related with genotypes of E. granulosus, in humans, without language neither genotyped method restriction, published between 1990-2019 were included. A systematic in WoS, EMBASE, MEDLINE, SCOPUS, Trip Database, BIREME, SciELO, LILACS, IBECS, and PAHO-WHO was carried out. In study variables were year of publication, country, number of samples, host and parasite organs, genotype identified, molecular marker and genes. Descriptive statistics were applied. Results: 701 related articles were identified; 62 fulfilled selection criteria, representing 1,511 samples. The existing evidence was published between 1994 and 2019; and mainly comes from Iran (45.2%). The most commonly used sequencing method was PCR amplification and Sanger type sequencing with partial or total genotyping of the cox1 gene. Genotyped method most frequently used was cox1 (79,0%). Genotypes most frequently identified were G1 and G1/G3 complex (49.1% and 32.2%). Conclusions: Publications related to genotypes of Eg in humans are scarce, heterogeneous, and presenting differing results. Eg G1/G3 accounts for most of the global burden worldwide.
Asunto(s)
Humanos , Animales , Echinococcus granulosus/genética , Equinococosis , Filogenia , Reacción en Cadena de la Polimerasa , GenotipoRESUMEN
Echinococcus granulosus (Eg) is a parasite causing an incidental infection in humans. The disease can be observed as a hydatid cyst in the liver and lungs and is found in dogs and wolves as definitive hosts. It is considered a neglected infectious disease which has rarely been studied in Mexico1.
Asunto(s)
Echinococcus granulosus/genética , Haplotipos , AnimalesRESUMEN
Cystic echinococcosis (CE) is a major neglected tropical zoonotic disease caused by the tissue-dwelling larval stage of the cestode parasite Echinococcus granulosus. For individuals suspected of CE, the diagnostic standard is imaging using ultrasonography, X rays, or computed tomography. These resource-demanding and expensive procedures are rarely available in endemic rural areas where CE is most prevalent. There is a critical need for a new approach to identify CE patients so that they can be managed early in the course of their infection. This study reports on the results of a diagnostic approach that identifies E. granulosus-derived cell-free DNA (cfDNA) in the urine of CE patients. Utilizing PCR to amplify a fragment of a major tandem repeat element found in E. granulosus nuclear DNA, urine samples from all seven imaging-confirmed CE patients who harbored active liver cysts were positive. In addition, the urine samples from 2/4 patients who presented with non-viable/calcified liver cysts were also PCR positive for the repeat fragment. To our knowledge, this is the first report of using parasite cfDNA from urine to diagnose CE. This approach provides an easy to implement and cost-effective method to survey for the prevalence of E. granulosus in humans populations.