Pathogens

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18 pages, 2324 KiB

Open AccessArticle

Cystic Echinococcosis: Clinical, Immunological, and Biomolecular Evaluation of Patients from Sardinia (Italy)

by Cinzia Santucciu, Piero Bonelli, Angela Peruzzu, Alessandro Fancellu, Vincenzo Marras, Antonello Carta, Scilla Mastrandrea, Giorgio Bagella, Toni Piseddu, Stefano Profili, Alberto Porcu and Giovanna Masala

Pathogens 2020, 9(11), 907; https://doi.org/10.3390/pathogens9110907 - 30 Oct 2020

Cited by 10 | Viewed by 3344

Abstract

Cystic echinococcosis (CE), a zoonotic disease caused by the larval stage of the tapeworm Echinococcus granulosus sensu lato (s.l.), is a worldwide public health problem. Echinococcus granulosus sensu stricto (s.s.), associated with G1 and G3 genotypes, is endemic with [...] Read more.

Cystic echinococcosis (CE), a zoonotic disease caused by the larval stage of the tapeworm Echinococcus granulosus sensu lato (s.l.), is a worldwide public health problem. Echinococcus granulosus sensu stricto (s.s.), associated with G1 and G3 genotypes, is endemic with high prevalence in the Mediterranean basin. The parasite’s life cycle comprises definitive hosts (canids) and intermediate hosts (ruminants) and can occasionally involve humans. The main aim of this research was to confirm the diagnosis of 13 patients suspected of CE who presented different complications and needed the surgical removal of the cysts. We also wanted to understand and clarify more the diagnosis of echinococcosis in humans. For this purpose, the patients first underwent cyst evaluation by ultrasound (US), immunological analysis, and then total pericystectomy, followed by parasitological, histopathological, and molecular biology examinations of the cysts. US stadiated one CE1, one CE2, eight CE3b, one CE4, and two CE5; immunology evidenced nine positives; histopathology confirmed 11 CE cysts, of which 8 fertile presenting protoscoleces were identified as E. granulosus s.s. by molecular biology, genotyped as three G1 and four G3 by neighbor-joining (NJ) phylogenetic tree. In conclusion, the results showed that 11 patients were affected by E. granulosus s.s. G1 orG3, and 2 cystic neoformations were of non-parasitic origin. Full article

(This article belongs to the Special Issue Echinococcus)

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9 pages, 983 KiB

Open AccessArticle

Bayesian Analysis of Three Methods for Diagnosis of Cystic Echinococcosis in Sheep

by Piero Bonelli, Federica Loi, Maria Giovanna Cancedda, Angela Peruzzu, Elisabetta Antuofermo, Elisabetta Pintore, Toni Piseddu, Giovanni Garippa and Giovanna Masala

Pathogens 2020, 9(10), 796; https://doi.org/10.3390/pathogens9100796 - 27 Sep 2020

Cited by 2 | Viewed by 2210

Abstract

Diagnosis of cystic echinococcosis (CE) in sheep is essentially based on necropsy findings. Clinical symptoms can be easily overlooked, while the use of immunological tests is still not recommended for an intra vitam diagnosis. This study assessed the performances of three post-mortem laboratory [...] Read more.

Diagnosis of cystic echinococcosis (CE) in sheep is essentially based on necropsy findings. Clinical symptoms can be easily overlooked, while the use of immunological tests is still not recommended for an intra vitam diagnosis. This study assessed the performances of three post-mortem laboratory methods in the diagnosis of ovine CE. In the absence of a single and accurate test as a gold standard, the results of multiple analytical tests can be combined to estimate diagnostic performance based on a Bayesian statistical approach. For this purpose, livers (n = 77), and lungs (n = 79) were sampled from adult sheep and examined using gross pathology, histopathology and molecular analyses. Data from the three diagnostic methods were analyzed using a Bayesian latent class analysis model to evaluate their diagnostic accuracy in terms of sensitivity (Se), specificity (Sp), positive predictive value (PPV) and negative predictive value (NPV). The gross pathology examination revealed excellent diagnostic capabilities in diagnosing ovine CE with an Se of 99.7 (96.7–99.8), Sp of 97.5 (90.3–99.8), PPV of 97.6 (90.5–100), and NPV of 99.7 (96.5–100). The experimental design used in this work could be implemented as a validation protocol in a quality assurance system. Full article

(This article belongs to the Special Issue Echinococcus)

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17 pages, 3246 KiB

Open AccessEditor’s ChoiceArticle

Species Detection within the Echinococcus granulosus sensu lato Complex by Novel Probe-Based Real-Time PCRs

by Pavlo Maksimov, Hannes Bergmann, Marion Wassermann, Thomas Romig, Bruno Gottstein, Adriano Casulli and Franz J. Conraths

Pathogens 2020, 9(10), 791; https://doi.org/10.3390/pathogens9100791 - 26 Sep 2020

Cited by 15 | Viewed by 3362

Abstract

Infections with eggs of Echinococcus granulosus sensu lato (s.l.) can cause cystic echinococcosis in intermediate host animals and humans. Upon ingestion of viable eggs, oncospheres hatch from the eggs and subsequently develop into fluid-filled larval cysts, most frequently in the liver [...] Read more.

Infections with eggs of Echinococcus granulosus sensu lato (s.l.) can cause cystic echinococcosis in intermediate host animals and humans. Upon ingestion of viable eggs, oncospheres hatch from the eggs and subsequently develop into fluid-filled larval cysts, most frequently in the liver or the lungs. The slowly growing cysts progressively interfere with organ function. The risk of infection is determined by the host range of the parasite, its pathogenicity and other epidemiologically relevant parameters, which differ significantly among the five species within the E. granulosus s.l. complex. It is therefore essential to diagnose the correct species within E. granulosus s.l. to help understand specific disease epidemiology and to facilitate effective implementation of control measures. For this purpose, simple, fast and cost-effective typing techniques are needed. We developed quantitative real-time polymerase chain reactions (qPCRs) to target polymorphic regions in the mitochondrial genome of E. granulosus s.l. In a single-step typing approach, we distinguished E. granulosus s.l. members in four epidemiologically relevant subgroups. These were E. granulosus sensu stricto, E. equinus, E. ortleppi and the E. canadensis cluster. The technique also allowed identification and differentiation of these species from other Echinococcus or Taenia taxa for samples isolated from cysts or faeces. Full article

(This article belongs to the Special Issue Echinococcus)

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12 pages, 1905 KiB

Open AccessArticle

Transcriptomic Analysis of the Early Strobilar Development of Echinococcus granulosus

by João Antonio Debarba, Martín Pablo Cancela Sehabiague, Karina Mariante Monteiro, Alexandra Lehmkuhl Gerber, Ana Tereza Ribeiro Vasconcelos, Henrique Bunselmeyer Ferreira and Arnaldo Zaha

Pathogens 2020, 9(6), 465; https://doi.org/10.3390/pathogens9060465 - 12 Jun 2020

Cited by 6 | Viewed by 2362

Abstract

Echinococcus granulosus has a complex life cycle involving two mammalian hosts. The transition from one host to another is accompanied by changes in gene expression, and the transcriptional events that underlie this transition have not yet been fully characterized. In this study, RNA-seq [...] Read more.

Echinococcus granulosus has a complex life cycle involving two mammalian hosts. The transition from one host to another is accompanied by changes in gene expression, and the transcriptional events that underlie this transition have not yet been fully characterized. In this study, RNA-seq was used to compare the transcription profiles of samples from E. granulosus protoscoleces induced in vitro to strobilar development at three time points. We identified 818 differentially expressed genes, which were divided into eight expression clusters formed over the entire 24 h period. An enrichment of gene transcripts with molecular functions of signal transduction, enzymes, and protein modifications was observed upon induction and developmental progression. This transcriptomic study provides insights for understanding the complex life cycle of E. granulosus and contributes for searching for the key genes correlating with the strobilar development, which can be used to identify potential candidates for the development of anthelmintic drugs. Full article

(This article belongs to the Special Issue Echinococcus)

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13 pages, 762 KiB

Open AccessArticle

Microsatellite Investigations of Multiple Echinococcus granulosus Sensu Stricto Cysts in Single Hosts Reveal Different Patterns of Infection Events between Livestock and Humans

by Selim M’rad, Myriam Oudni-M’rad, Vanessa Bastid, Laure Bournez, Sana Mosbahi, Abdelallatif Nouri, Hamouda Babba, Frédéric Grenouillet, Franck Boué and Gérald Umhang

Pathogens 2020, 9(6), 444; https://doi.org/10.3390/pathogens9060444 - 5 Jun 2020

Cited by 6 | Viewed by 2945

Abstract

Cystic echinococcosis (CE) caused by the cestode Echinococcus granulosus sensu lato (s.l.) is a worldwide zoonosis and E. granulosus sensu stricto (s.s.) is the most common species associated with animal and human diseases. The objective of this study was to obtain a better [...] Read more.

Cystic echinococcosis (CE) caused by the cestode Echinococcus granulosus sensu lato (s.l.) is a worldwide zoonosis and E. granulosus sensu stricto (s.s.) is the most common species associated with animal and human diseases. The objective of this study was to obtain a better understanding of CE infection in livestock and humans from very low and high endemic areas—France and Tunisia—by studying the genetic diversity of E. granulosus s.s. at the intra-individual host level. This genetic diversity was studied using EgSca6 and EgSca11 microsatellite profiles in 93 sheep from France and Tunisia, and in 12 cattle and 31 children from Tunisia only, all presenting multiple CE cysts (2 to 10 cysts). Overall, 96% of sheep, 92% of cattle, and 48% of children had at least two cysts with different microsatellite profiles. Inversely, 35% of sheep, 17% of cattle, and 65% of children had at least two cysts with the same microsatellite profile. The genotyping results for the CE samples highlight high and similar genetic diversity in France and Tunisia, suggesting that the probability of being successively infected by CE of the same microsatellite profile was rare in both countries. Therefore, our results suggest that in rare cases, several eggs of the same microsatellite profile, from two to seven in our data, can be ingested simultaneously in a single infection event and develop into several cysts in livestock and children. They also indicate that multiple infection events are frequent in livestock, even in a low endemic country such as France, and are less frequent but not negligible in children in a high endemic country such as Tunisia. Moreover, this is the first time that genetic evidence of secondary CE has been found. Further studies are needed to better assess the pattern of infection events in livestock and humans, especially by studying the genetic diversity of adult worms in definitive hosts. Full article

(This article belongs to the Special Issue Echinococcus)

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21 pages, 3475 KiB

Open AccessArticle

Genotyping Echinococcus multilocularis in Human Alveolar Echinococcosis Patients: An EmsB Microsatellite Analysis

by Jenny Knapp, Bruno Gottstein, Stéphane Bretagne, Jean-Mathieu Bart, Gérald Umhang, Carine Richou, Solange Bresson-Hadni and Laurence Millon

Pathogens 2020, 9(4), 282; https://doi.org/10.3390/pathogens9040282 - 13 Apr 2020

Cited by 17 | Viewed by 3081

Abstract

For clinical epidemiology specialists, connecting the genetic diversity of Echinococcus multilocularis to sources of infection or particular sites has become somewhat of a holy grail. It is very difficult to trace the infection history of alveolar echinococcosis (AE) patients as there may be [...] Read more.

For clinical epidemiology specialists, connecting the genetic diversity of Echinococcus multilocularis to sources of infection or particular sites has become somewhat of a holy grail. It is very difficult to trace the infection history of alveolar echinococcosis (AE) patients as there may be an incubation period of five to 15 years before reliable diagnosis. Moreover, the variability of parasitic manifestations in human patients raises the possibility of genetically different isolates of E. multilocularis having different levels of pathogenicity. Thus, the exposure of human patients to different strains or genotypes circulating in geographically different environments may lead to different disease outcomes. Molecular tools, such as the microsatellite marker EmsB, were required to investigate these aspects. This genetic marker was previously tested on a collection of 1211 European field samples predominantly of animal origin, referenced on a publicly available database. In this study, we investigated a panel of 66 metacestode samples (between 1981 and 2019) recovered surgically from 63 patients diagnosed with alveolar echinococcosis originating from four European countries (France, Switzerland, Germany, Belgium). In this study, we identified nine EmsB profiles, five of which were found in patients located in the same areas of France and Switzerland. One profile was detected on both sides of the French-Swiss border, whereas most patients from non-endemic regions clustered together in another profile. EmsB profiles appeared to remain stable over time because similar profiles were detected in patients who underwent surgery recently and patients who underwent surgery some time ago. This study sheds light on possible pathways of contamination in humans, including proximity contamination in some cases, and the dominant contamination profiles in Europe, particularly for extrahepatic lesions. Full article

(This article belongs to the Special Issue Echinococcus)

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Review

Jump to: Research, Other

9 pages, 209 KiB

Open AccessEditor’s ChoiceReview

The Molecular Epidemiology of Echinococcus Infections

by R. C. Andrew Thompson

Pathogens 2020, 9(6), 453; https://doi.org/10.3390/pathogens9060453 - 8 Jun 2020

Cited by 25 | Viewed by 4070

Abstract

Molecular epidemiology (ME) is the application of molecular tools to determine the causation of disease. With infectious diseases, such as echinococcosis, this applies to identifying and characterising the aetiological agents and elucidating host range. Such an approach has been very successful with the [...] Read more.

Molecular epidemiology (ME) is the application of molecular tools to determine the causation of disease. With infectious diseases, such as echinococcosis, this applies to identifying and characterising the aetiological agents and elucidating host range. Such an approach has been very successful with the causative agents of echinococcosis, species of Echinococcus, initially by providing a workable and practical taxonomy and subsequently determining transmission patterns in endemic areas. This review summarises the taxonomy and nomenclature of species of Echinococcus and provides an update on ME investigations of the ecology of Echinococcus transmission, particularly in areas where more than one species of Echinococcus is maintained in cycles of transmission that may interact. Full article

(This article belongs to the Special Issue Echinococcus)

Other

Jump to: Research, Review

11 pages, 1615 KiB

Open AccessBrief Report

In-House Developed ELISA Indicates High Prevalence of Anti-Echinococcus granulosus IgG in Sheep Population—An Update from Pakistan

by Mughees Aizaz Alvi, John Asekhaen Ohiolei, Li Li, Muhammad Saqib, Muhammad Hammad Hussain, Muhammad Haleem Tayyab, Muzafar Ghafoor, Warda Qamar, Noman Yousaf Faridi, Anum Aizaz Alvi, Bao-Quan Fu, Hong-Bin Yan and Wan-Zhong Jia

Pathogens 2020, 9(11), 905; https://doi.org/10.3390/pathogens9110905 - 29 Oct 2020

Cited by 3 | Viewed by 2227

Abstract

Cystic echinococcosis (CE) is a World Health Organization (WHO)-listed neglected tropical farm economy jeopardizing and public health concern disease. This study was aimed at furnishing sero-epidemiological baseline data of CE in sheep in Pakistan, where data are non-existent. For this purpose, two sheep-rich [...] Read more.

Cystic echinococcosis (CE) is a World Health Organization (WHO)-listed neglected tropical farm economy jeopardizing and public health concern disease. This study was aimed at furnishing sero-epidemiological baseline data of CE in sheep in Pakistan, where data are non-existent. For this purpose, two sheep-rich provinces of Pakistan were selected, and 728 sheep sera were collected using probability proportional to size (PPS) statistical technique. Epidemiological information was recorded on a questionnaire for the estimation of potential risk factors. The serum samples were analyzed for IgG antibodies against Echinococcus granulosus using an in-house-developed EgAgB-based ELISA kit. The overall seroprevalence recorded was 21.98% (160/728) in the tested sheep, suggesting higher seropositivity in sheep from Punjab (23.73%) as compared to Khyber Pakhtunkhwa (KPK) (19.04%). The overall apparent prevalence observed by this ELISA method was almost similar to the calculated true prevalence (21.77%). Prevalence was significantly different (p < 0.05) among sheep from different districts. Higher prevalence was found in females (22.54%, OR 1.41), age group > 5 years (29.66%, OR 1.64), crossbreeds (42.85%, OR 2.70), and sheep with pasture access (25.96%, OR 3.06). Being in age group > 5 years and having pasture access were the factors significantly associated with seropositivity (p < 0.05). This study provides serological evidence of E. granulosus infection in sheep and can be used as a model for ante-mortem screening of the sheep globally. Full article

(This article belongs to the Special Issue Echinococcus)

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