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      Molecular identification of Fasciola spp. (Digenea: Fasciolidae) in Egypt Translated title: Caractérisation moléculaire de Fasciola spp. en Egypte

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          Abstract

          A total of 134 Egyptian liver flukes were collected from different definitive hosts (cattle, sheep, and buffaloes) to identify them via the use of PCR-RFLP and sequence analysis of the first nuclear ribosomal internal transcribed spacer (ITS1). Specimens of F. hepatica from France, as well as F. gigantica from Cameroon were included in the study for comparison. PCR products of ITS1 were subjected for digestion by RsaI restriction enzyme and visualized on agarose gel. According to RFLP pattern, Egyptian flukes were allocated into two categories. The first was identical to that of French hepatica flukes to have a pattern of 360, 100, and 60 (bp) band size, whereas the second resembled to that of Cameroonian gigantica worms to have a profile of 360, 170, and 60 bp in size. Results of RFLP analysis were confirmed by sequence analysis of representative ITS1 amplicons. No hybrid forms were detected in the present study. Taken together, this study concluded that both species of Fasciola are present in Egypt, whereas the hybrid form may be not very common.

          Translated abstract

          Des douves égyptiennes (134 spécimens) provenant de différents hôtes définitifs (bovins, buffles et moutons) ont été analysées par PCR-RFLP et l’étude de la séquence ITS-1 pour identifier les espèces locales de Fasciola ( F. hepatica, F. gigantica, ou les formes hybrides entre ces deux espèces). Des douves provenant de France ( F. hepatica) et du Cameroun ( F. gigantica) ont été utilisées comme références. Deux types de bandes ont été trouvés dans les fragments de la séquence ITS-1 : le premier type est identique à celui des F. hepatica de France (trois bandes de 60, 100 et 360 bp) tandis que le second ressemble à celui des F. gigantica du Cameroun (trois bandes de 60, 170 et 360 bp). De plus, le séquençage des amplicons ITS-1 confirme cette différence en montrant la présence de six sites nucléotidiques variables, ce qui permet de discriminer F. hepatica de F. gigantica. Aucune forme intermédiaire entre les deux Fasciola n’a été trouvée dans les spécimens analysés. Pris ensemble, cette étude permet de conclure que les deux espèces de Fasciola sont présentes en Égypte, alors que la forme hybride pourrait être pas très commune.

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          Most cited references 19

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          Fascioliasis and other plant-borne trematode zoonoses.

          Fascioliasis and other food-borne trematodiases are included in the list of important helminthiases with a great impact on human development. Six plant-borne trematode species have been found to affect humans: Fasciola hepatica, Fasciola gigantica and Fasciolopsis buski (Fasciolidae), Gastrodiscoides hominis (Gastrodiscidae), Watsonius watsoni and Fischoederius elongatus (Paramphistomidae). Whereas F. hepatica and F. gigantica are hepatic, the other four species are intestinal parasites. The fasciolids and the gastrodiscid cause important zoonoses distributed throughout many countries, while W. watsoni and F. elongatus have been only accidentally detected in humans. Present climate and global changes appear to increasingly affect snail-borne helminthiases, which are strongly dependent on environmental factors. Fascioliasis is a good example of an emerging/re-emerging parasitic disease in many countries as a consequence of many phenomena related to environmental changes as well as man-made modifications. The ability of F. hepatica to spread is related to its capacity to colonise and adapt to new hosts and environments, even at the extreme inhospitality of very high altitude. Moreover, the spread of F. hepatica from its original European range to other continents is related to the geographic expansion of its original European lymnaeid intermediate host species Galba truncatula, the American species Pseudosuccinea columella, and its adaptation to other lymnaeid species authochthonous in the newly colonised areas. Although fasciolopsiasis and gastrodiscoidiasis can be controlled along with other food-borne parasitoses, fasciolopsiasis still remains a public health problem in many endemic areas despite sustained WHO control programmes. Fasciolopsiasis has become a re-emerging infection in recent years and gastrodiscoidiasis, initially supposed to be restricted to Asian countries, is now being reported in African countries.
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            Chapter 2. Fasciola, lymnaeids and human fascioliasis, with a global overview on disease transmission, epidemiology, evolutionary genetics, molecular epidemiology and control.

            Fascioliasis, caused by liver fluke species of the genus Fasciola, has always been well recognized because of its high veterinary impact but it has been among the most neglected diseases for decades with regard to human infection. However, the increasing importance of human fascioliasis worldwide has re-launched interest in fascioliasis. From the 1990s, many new concepts have been developed regarding human fascioliasis and these have furnished a new baseline for the human disease that is very different to a simple extrapolation from fascioliasis in livestock. Studies have shown that human fascioliasis presents marked heterogeneity, including different epidemiological situations and transmission patterns in different endemic areas. This heterogeneity, added to the present emergence/re-emergence of the disease both in humans and animals in many regions, confirms a worrying global scenario. The huge negative impact of fascioliasis on human communities demands rapid action. When analyzing how better to define control measures for endemic areas differing at such a level, it would be useful to have genetic markers that could distinguish each type of transmission pattern and epidemiological situation. Accordingly, this chapter covers aspects of aetiology, geographical distribution, epidemiology, transmission and control in order to obtain a solid baseline for the interpretation of future results. The origins and geographical spread of F. hepatica and F. gigantica in both the ruminant pre-domestication times and the livestock post-domestication period are analyzed. Paleontological, archaeological and historical records, as well as genetic data on recent dispersal of livestock species, are taken into account to establish an evolutionary framework for the two fasciolids across all continents. Emphasis is given to the distributional overlap of both species and the roles of transportation, transhumance and trade in the different overlap situations. Areas with only one Fasciola spp. are distinguished from local and zonal overlaps in areas where both fasciolids co-exist. Genetic techniques applied to liver flukes in recent years that are useful to elucidate the genetic characteristics of the two fasciolids are reviewed. The intra-specific and inter-specific variabilities of 'pure'F. hepatica and 'pure'F. gigantica were ascertained by means of complete sequences of ribosomal deoxyribonucleic acid (rDNA) internal transcribed spacer (ITS)-2 and ITS-1 and mitochondrial deoxyribonucleic acid (mtDNA) cox1 and nad1 from areas with only one fasciolid species. Fasciolid sequences of the same markers scattered in the literature are reviewed. The definitive haplotypes established appear to fit the proposed global evolutionary scenario. Problems posed by fasciolid cross-breeding, introgression and hybridization in overlap areas are analyzed. Nuclear rDNA appears to correlate with adult fluke characteristics and fasciolid/lymnaeid specificity, whereas mtDNA does not. However, flukes sometimes appear so intermediate that they cannot be ascribed to either F. hepatica-like or F. gigantica-like forms and snail specificity may be opposite to the one deduced from the adult morphotype. The phenotypic characteristics of adults and eggs of 'pure'F. hepatica and F. gigantica, as well as of intermediate forms in overlap areas, are compared, with emphasis on the definitive host influence on egg size in humans. Knowledge is sufficient to support F. hepatica and F. gigantica as two valid species, which recently diverged by adaptation to different pecoran and lymnaeid hosts in areas with differing environmental characteristics. Their phenotypic differences and ancient pre-domestication origins involve a broad geographical area that largely exceeds the typical, more local scenarios known for sub-species units. Phenomena such as abnormal ploidy and aspermic parthenogenesis in hybrids suggest that their separate evolution in pre-domestication times allowed them to achieve almost total genetic isolation. Recent sequencing results suggest that present assumptions on fasciolid-lymnaeid specificity might be wrong. The crucial role of lymnaeids in fascioliasis transmission, epidemiology and control was the reason for launching a worldwide lymnaeid molecular characterization initiative. This initiative has already furnished useful results on several continents. A standardized methodology for fasciolids and lymnaeids is proposed herein in order that future work is undertaken on a comparable basis. A complete understanding of molecular epidemiology is expected to help greatly in designing global actions and local interventions for control of fascioliasis.
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              Epidemiology of human fascioliasis: a review and proposed new classification.

              The epidemiological picture of human fascioliasis has changed in recent years. The number of reports of humans infected with Fasciola hepatica has increased significantly since 1980 and several geographical areas have been described as endemic for the disease in humans, with prevalence and intensity ranging from low to very high. High prevalence of fascioliasis in humans does not necessarily occur in areas where fascioliasis is a major veterinary problem. Human fascioliasis can no longer be considered merely as a secondary zoonotic disease but must be considered to be an important human parasitic disease. Accordingly, we present in this article a proposed new classification for the epidemiology of human fascioliasis. The following situations are distinguished: imported cases; autochthonous, isolated, nonconstant cases; hypo-, meso-, hyper-, and holoendemics; epidemics in areas where fascioliasis is endemic in animals but not humans; and epidemics in human endemic areas.
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                Author and article information

                Journal
                Parasite
                Parasite
                parasite
                Parasite : journal de la Société Française de Parasitologie
                EDP Sciences
                1252-607X
                1776-1042
                May 2012
                15 May 2012
                : 19
                : 2 ( publisher-idID: parasite/2012/02 )
                : 177-182
                Affiliations
                [1 ] Zoology Department, Faculty of Science, Tanta University Egypt
                [2 ] Department of Zoology, Faculty of Science, Kafr El-Sheikh University Kafr El Sheikh 33516 Egypt
                [3 ] EA 3174 Tropical and Comparative Neuroepidemiology, Institute of Tropical Neurology, and IFR 145, GEIST, Faculties of Medicine and Pharmacy, University of Limoges France
                Author notes
                [* ]Correspondence: Said Amer. Tel./Fax: 2 047-3215176. E-mail: mssamer5@ 123456yahoo.com
                Article
                parasite2012192p177 10.1051/parasite/2012192177
                10.1051/parasite/2012192177
                3671433
                22550630
                © PRINCEPS Editions, Paris, 2012

                This is an Open Access article distributed under the terms of the Creative Commons Attribution License ( http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

                Page count
                Figures: 3, Tables: 1, Equations: 0, References: 31, Pages: 6
                Categories
                Research Note

                pcr-rflp, egypt, fasciola gigantica, fasciola hepatica, égypte

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