RESUMEN
BACKGROUND: Snails of the Lymnaeidae family are the intermediate hosts of Fasciola species, the causative agents of fascioliasis. The purpose of this study was to determine the prevalence of Fasciola species in lymnaeid snails and to investigate the association of geoclimatic factors and Fasciola species distribution in northwestern provinces of Iran using geographical information system (GIS) data. METHODS: A total of 2000 lymnaeid snails were collected from 33 permanent and seasonal habitats in northwestern Iran during the period from June to November 2021. After identification by standard morphological keys, they were subjected to shedding and crushing methods. Different stages of Fasciola obtained from these snails were subjected to the ITS1 polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method for species identification. The associations of weather temperature, rainfall, humidity, evaporation, air pressure, wind speed, elevation, and land cover with the distribution of Fasciola species were investigated. Geographical and statistical analysis was performed using ArcMap and SPSS software, respectively, to determine factors related to Fasciola species distribution. RESULTS: Of the 2000 snails collected, 19 were infected with Fasciola hepatica (0.09%), six with F. gigantica (0.03%), and 13 with other trematodes. Among geoclimatic and environmental factors, mean humidity, maximum humidity, and wind speed were significantly higher in areas where F. hepatica was more common than F. gigantica. The altitude of F. hepatica-prevalent areas was generally lower than F. gigantica areas. No significant relationship was observed between other investigated geoclimatic factors and the distribution of infected snails. CONCLUSIONS: The present study showed the relationship of humidity and wind speed with the distribution of snails infected with F. hepatica or F. gigantica in the northwestern regions of Iran. In contrast to F. gigantica, F. hepatica was more prevalent in low-altitude areas. Further research is recommended to elucidate the relationship between geoclimatic factors and the presence of intermediate hosts of the two Fasciola species.
Asunto(s)
Fasciola , Fascioliasis , Caracoles , Animales , Irán/epidemiología , Fascioliasis/epidemiología , Fascioliasis/veterinaria , Fascioliasis/parasitología , Caracoles/parasitología , Fasciola/genética , Fasciola/aislamiento & purificación , Fasciola/clasificación , Fasciola hepatica/genética , Fasciola hepatica/aislamiento & purificación , Fasciola hepatica/fisiología , Fasciola hepatica/clasificación , Clima , Ecosistema , Estaciones del Año , Polimorfismo de Longitud del Fragmento de RestricciónRESUMEN
Fasciola gigantica is a widespread parasite that causes neglected disease in livestock worldwide. Its high transmissibility and dispersion are attributed to its ability to infect intermediate snail hosts and adapt to various mammalian definitive hosts. This study investigated the variation and population dynamics of F. gigantica in cattle, sheep, and goats from three states in Sudan. Mitochondrial cytochrome c oxidase subunit I (COI) and NADH dehydrogenase subunit 1 (ND1) genes were sequenced successfully to examine intra and interspecific differences. ND1 exhibited higher diversity than COI, with 15 haplotypes and 10 haplotypes, respectively. Both genes had high haplotype diversity but low nucleotide diversity, with 21 and 11 polymorphic sites for ND1 and COI, respectively. Mismatch distribution analysis and neutrality tests revealed that F. gigantica from different host species was in a state of population expansion. Maximum likelihood phylogenetic trees and median networks revealed that F. gigantica in Sudan and other African countries had host-specific and country-specific lineages for both genes. The study also indicated that F. gigantica-infected small ruminants were evolutionarily distant, suggesting deep and historical interspecies adaptation.
Asunto(s)
Complejo IV de Transporte de Electrones , Fasciola , Fascioliasis , Variación Genética , Cabras , Haplotipos , NADH Deshidrogenasa , Filogenia , Dinámica Poblacional , Animales , Sudán/epidemiología , Fasciola/genética , Fasciola/clasificación , Fasciola/aislamiento & purificación , Fascioliasis/veterinaria , Fascioliasis/parasitología , Fascioliasis/epidemiología , Ovinos/parasitología , Cabras/parasitología , Bovinos , NADH Deshidrogenasa/genética , Complejo IV de Transporte de Electrones/genética , Enfermedades de las Cabras/parasitología , Enfermedades de las Cabras/epidemiología , Rumiantes/parasitología , Enfermedades de las Ovejas/parasitología , Enfermedades de las Ovejas/epidemiología , Enfermedades de los Bovinos/parasitología , Enfermedades de los Bovinos/epidemiología , Análisis de Secuencia de ADNRESUMEN
PURPOSE: The objective of this study is to study the secondary structure analysis of Fasciola flukes from a rare mithun host from Manipur. Fascioliasis, a neglected tropical trematodiasis, is poorly studied in India and is widely believed to be predominantly caused by F. gigantica. Through this study, we want to assess the flukes from the rare semi-wild ruminants of Northeast India. This study is important as the mithun population is semi-wild and its population is declining in Manipur. METHODS: Sample collected from the difficult and challenging terrain of Northeast India. The sample was collected from mithun and observed under the microscope. DNA was isolated, sequenced, and analyzed using various bioinformatics tools. The secondary structure analysis of the Internal Transcribed Spacer 2 (ITS2) region was also performed. RESULTS: The secondary structure species tree corroborated the Bayesian inference and, hence, strengthened the phylogeny reconstructed. The annotated ITS2 sequence and RNA secondary of the Manipur isolate displayed the typical four-helix or four-domain model. Helix III reveals the presence of the UGGU motif with other deviations like UGG and GGU. CONCLUSION: This is an in-depth analysis of the secondary structure of Fasciola species. The present study has demonstrated the usefulness of ITS2 and its secondary structures for characterizing parasites. The information on fascioliasis in the mithun's population presents itself useful with regards to their conservation strategy as their populations in both Manipur and Nagaland are dwindling.
Asunto(s)
Fasciola , Fascioliasis , Conformación de Ácido Nucleico , Filogenia , Rumiantes , Animales , India/epidemiología , Fasciola/genética , Fasciola/clasificación , Fasciola/aislamiento & purificación , Fascioliasis/veterinaria , Fascioliasis/parasitología , Fascioliasis/epidemiología , Rumiantes/parasitología , ADN de Helmintos/genética , ADN de Helmintos/química , ADN Espaciador Ribosómico/genética , ADN Espaciador Ribosómico/química , Análisis de Secuencia de ADNRESUMEN
Fasciola gigantica and Fasciola hepatica are causative pathogens of fascioliasis, with the widest latitudinal, longitudinal, and altitudinal distribution; however, among parasites, they have the largest sequenced genomes, hindering genomic research. In the present study, we used various sequencing and assembly technologies to generate a new high-quality Fasciola gigantica reference genome. We improved the integration of gene structure prediction, and identified two independent transposable element expansion events contributing to (1) the speciation between Fasciola and Fasciolopsis during the Cretaceous-Paleogene boundary mass extinction, and (2) the habitat switch to the liver during the Paleocene-Eocene Thermal Maximum, accompanied by gene length increment. Long interspersed element (LINE) duplication contributed to the second transposon-mediated alteration, showing an obvious trend of insertion into gene regions, regardless of strong purifying effect. Gene ontology analysis of genes with long LINE insertions identified membrane-associated and vesicle secretion process proteins, further implicating the functional alteration of the gene network. We identified 852 predicted excretory/secretory proteins and 3300 protein-protein interactions between Fasciola gigantica and its host. Among them, copper/zinc superoxide dismutase genes, with specific gene copy number variations, might play a central role in the phase I detoxification process. Analysis of 559 single-copy orthologs suggested that Fasciola gigantica and Fasciola hepatica diverged at 11.8 Ma near the Middle and Late Miocene Epoch boundary. We identified 98 rapidly evolving gene families, including actin and aquaporin, which might explain the large body size and the parasitic adaptive character resulting in these liver flukes becoming epidemic in tropical and subtropical regions.
Asunto(s)
Enfermedades de los Bovinos/parasitología , Elementos Transponibles de ADN , Evolución Molecular , Fasciola/genética , Fascioliasis/veterinaria , Genoma de los Helmintos , Adaptación Fisiológica , Animales , Búfalos/parasitología , Bovinos , Fasciola/clasificación , Fasciola/fisiología , Fascioliasis/parasitología , Fascioliasis/fisiopatología , Genómica , Proteínas del Helminto/genética , Interacciones Huésped-Parásitos , FilogeniaRESUMEN
This study aimed to identify species of Fasciola flukes in Dali Prefecture (Yunnan Province, China) and analyze their genetic diversity. Fasciola flukes (n = 122) were collected from cattle livers in a farmers' market in Xiaguan Town, Dali Prefecture. Nucleotide sequences of ribosomal internal transcribed spacer (ITS) as well as nicotinamide adenine dinucleotide dehydrogenase subunit 1 (ND1) and mitochondrial cytochrome c oxidase subunit 1 (CO1) were amplified, sequenced, and subjected to homology analysis. The heterozygosity ratios of different ITS alleles were determined using the peak-height ratio of heterozygous loci. Multiplex PCR analysis of the nuclear protein coding gene, phosphoenolpyruvate carboxykinase (pepck), was used to identify Fasciola species. Multiple ND1 sequence alignments enabled further genetic diversity analysis of regional Fasciola flukes. Seven ITS sequences belonged to F. hepatica and 115 belonged to Fh/Fg heterozygous flukes. Sequencing analysis of heterozygous flukes revealed 11 heterozygous loci with double peaks, with significantly variable ratios among individuals. ND1 and CO1 results indicated that one specimen was identical to F. hepatica, while 121 specimens were identical to F. gigantica or contained one variable site. Multiplex PCR results for pepck showed that double bands for F. hepatica and F. gigantica were amplified from Dali Fasciola specimens; hence, they were all heterozygous. By combining ITS, ND1, and CO1 sequences with multiplex pepck PCR results, all 122 specimens were identified as Fh/Fg heterozygous Fasciola flukes. Our experimental results preliminarily confirmed a high degree of Fh/Fg heterozygosity among Fasciola flukes in the Dali area. Selecting multiple molecular markers for concurrent analysis will provide more comprehensive and accurate genetic information.
Asunto(s)
Enfermedades de los Bovinos/parasitología , Fasciola/genética , Fascioliasis/veterinaria , Polimorfismo Genético , Animales , Secuencia de Bases , Bovinos , China , Fasciola/clasificación , Fasciola/aislamiento & purificación , Fascioliasis/parasitología , Proteínas del Helminto/análisis , Hepatopatías/parasitología , Hepatopatías/veterinaria , Proteínas Mitocondriales/análisis , NADH Deshidrogenasa/análisis , Alineación de Secuencia/veterinariaRESUMEN
Fasciolosis in ruminants is a relentless constraint in the livestock industry across the world. Immuno-prophylactic vaccines against fasciolosis may not come up in near future, rendering the control of this scourge with chemotherapy and snail population control. With the alarming threats of anti-fasciolid drug resistance reported from certain parts of the world; the control of fasciolosis should be directed towards the development of rapid and reliable diagnostic tools to execute the specific and discrete treatment. Understanding the epidemiology of Fasciola, its genomics and proteomics, host-parasite interplay, and advances in drug design research is vital for improving animal health that would ultimately succour to meet the ever-increasing demand for food. Due to possible differences in immune response depending on the species of the host and parasite, immuno-prophylactic studies in India should aim at achieving protective efficacy in buffalo against F. gigantica as workers from other countries concentrate primarily on vaccination of cattle and sheep against F. hepatica. This manuscript focused on the research that has been carried out in India for understanding the epidemiology, genetic diversity, immuno-diagnosis, and possible control measure in terms of immuno-prophylaxis and drug designing against tropical fasciolosis caused by Fasciola gigantica.
Asunto(s)
Fasciola/genética , Fascioliasis/epidemiología , Rumiantes/parasitología , Animales , Fasciola/clasificación , Fascioliasis/diagnóstico , Fascioliasis/prevención & control , Humanos , India/epidemiología , Prevalencia , Caracoles/parasitología , VacunasRESUMEN
All 225 Fasciola flukes obtained from domestic animals (73 cattle, 7 sheep and 1 pig) of 18 distinct geographic areas in Ecuador-South America, were identified as Fasciola hepatica, based on molecular analyses of nuclear pepck and pold genes, and mitochondrial nad1gene as well as the morphological observation of sperm within the seminal vesicles. Fasciola gigantica and parthenogenic Fasciola forms endemic to Asian countries were not found in this study, although zebu cattle and water buffalos have introduced into South America from Asia; this could be due to the absence of suitable intermediate host snails. The results of pepck analysis using multiplex PCR developed previously showed that 32 of the flukes could not be confirmed as F. hepatica, suggesting that the method is unreliable for the accurate discrimination of F. hepatica, and that pepck gene of the species consists of multiple loci, not a single locus. The results of genetic diversity, phylogenetic, and network analyses based on mitochondrial nad1 sequences suggest that F. hepatica populations in South America, including Ecuador, formed from the ancestral F. hepatica individuals introduced into the continent along with anthropogenic movement of livestock infected with the species.
Asunto(s)
Fasciola hepatica/clasificación , Variación Genética , Animales , Ecuador , Fasciola/clasificación , Fasciola/genética , Fasciola/aislamiento & purificación , Fasciola hepatica/genética , Fasciola hepatica/aislamiento & purificación , Proteínas del Helminto/análisis , Proteínas Mitocondriales/análisis , FilogeniaRESUMEN
Fascioliasis is a zoonotic infection of humans and, more commonly, ruminants. It is caused by 2 liver fluke species, Fasciola hepatica and Fasciola gigantica, which differ in size. The traditional morphological methods used to distinguish the 2 species can be unreliable, particularly in the presence of hybrids between the 2 species. The development of advanced molecular methods has allowed for more definitive identification of Fasciola species, including their hybrids. Hybrids are of concern, as it is thought that they could acquire advantageous traits such as increased pathogenicity and host range. In 2013, we collected flukes from Fasciola-positive cattle, sheep, and goats slaughtered in 4 Chadian abattoirs. DNA from 27 flukes was extracted, amplified, and analyzed to identify species using the ITS1+2 locus. Twenty-six of the 27 flukes were identified as F. gigantica, while the remaining fluke showed heterozygosity at all variable sites that distinguish F. hepatica and F. gigantica. Cloning and sequencing of both alleles confirmed the presence of 1 F. hepatica and 1 F. gigantica allele. To our knowledge, this is the first unambiguous, molecular demonstration of the presence of such a hybrid in a bovine in sub-Saharan Africa.
Asunto(s)
Enfermedades de los Bovinos/parasitología , Quimera/genética , Fasciola hepatica/genética , Fascioliasis/veterinaria , Mataderos , Animales , Bovinos , Chad , Quimera/clasificación , Secuencia de Consenso , Fasciola/clasificación , Fasciola/genética , Fasciola/aislamiento & purificación , Fasciola hepatica/clasificación , Fasciola hepatica/aislamiento & purificación , Fascioliasis/parasitología , Femenino , Inspección de Alimentos , Enfermedades de las Cabras/parasitología , Cabras , Polimorfismo de Nucleótido Simple/genética , Ovinos , Enfermedades de las Ovejas/parasitologíaRESUMEN
Fasciola spp. are responsible for over 3 billion US dollars of production loss annually in livestock and cause widespread zoonotic disease. Nevertheless, understating of the emergence and spread of the trematode species is poor. The multiplicity of F. gigantica infection and its spread is potentially influenced by multiple factors, including the abundance of suitable intermediate hosts, climatic conditions favouring the completion of the parasite's lifecycle, and translocation of infected animals, or free-living parasite stages between regions. Here we describe the development of a 'tremabiome' metabarcoding sequencing method to explore the numbers of F. gigantica genotypes per infection and patterns of parasite spread, based on genetic characteristics of the mitochondrial NADH dehydrogenase 1 (mt-ND-1) locus. We collected F. gigantica from three abattoirs in the Punjab and Balochistan provinces of Pakistan, and our results show a high level of genetic diversity in 20 F. gigantica populations derived from small and large ruminants consigned to slaughter in both provinces. This implies that F. gigantica can reproduce in its definitive hosts through meiosis involving cross- and self-breeding, as described in the closely related species, Fasciola hepatica. The genetic diversity between the 20 populations derived from different locations also illustrates the impact of animal movements on gene flow. Our results demonstrate the predominance of single haplotypes, consistent with a single introduction of F. gigantica infection in 85% of the hosts from which the parasite populations were derived. This is consistent with clonal reproduction in the intermediate snail hosts.
Asunto(s)
Búfalos , Enfermedades de los Bovinos/epidemiología , Fasciola/aislamiento & purificación , Fascioliasis/veterinaria , Variación Genética , Enfermedades de las Cabras/epidemiología , Enfermedades de las Ovejas/epidemiología , Animales , Bovinos , Enfermedades de los Bovinos/parasitología , Fasciola/clasificación , Fascioliasis/epidemiología , Fascioliasis/parasitología , Genotipo , Enfermedades de las Cabras/parasitología , Cabras , Pakistán/epidemiología , Ovinos , Enfermedades de las Ovejas/parasitologíaRESUMEN
Fasciolosis is a public health problem originally transmitted from livestock. Although molecular analysis of the nuclear and mitochondrial genes allow discrimination between the two known species of Fasciola, F. hepatica and F. gigantica, these markers do not permit the detection of hybrid forms. On the other hand, molecular analysis of the pepck and pold genes from Fasciola do permit the detection of hybrid isolates and this study has not yet been performed on specimens from human and domesticated animals in Iran. Therefore, the aim of this study was to molecularly analyze Fasciola isolates using both conventional and the two new genetic markers mentioned above. Fifty-three adult Fasciola worms were isolated from the livers of 28 domesticated animals, including sheep, cattle and goat. Moreover, an adult worm was obtained from the bile duct of an infected woman during endoscopic retrograde cholangio-pancratography (ERCP). Targeted fragments from the ITS-1, NDI and COX1 genes were amplified using specific primers. PCR products were sequenced and genetically analyzed using MEGA v.7 and DnaSP software. Additionally, the pepck and pold regions were amplified and analyzed using multiplex PCR and RLFP-PCR, respectively. Multiple alignment of sequenced fragments showed highest similarity among the ITS-1 sequences isolated from all four hosts in comparison to the other genes. Furthermore, sequence diversity across the COX1 was higher than the NDI and ITS-1. Diversity among sequences isolated from cattle was higher than those from sheep and goat in all three genes. Tajima's D and Fu's Fs were negative and statistically significant for all the genes except ITS-1. Phylogenetic trees showed that the human F. hepatica isolate was closer to sheep isolates. The results of the pepck and pold analyses showed that all isolates were F. hepatica and there were no hybrid forms among samples. The molecular analyses corroborated this finding.
Asunto(s)
Fasciola/clasificación , Fasciola/genética , Fascioliasis/epidemiología , Fascioliasis/parasitología , Genes de Helminto , Tipificación de Secuencias Multilocus , Animales , Variación Genética , Geografía Médica , Humanos , Irán/epidemiología , Pruebas de Neutralización , Filogenia , Reacción en Cadena de la Polimerasa , Polimorfismo de Longitud del Fragmento de Restricción , Vigilancia en Salud PúblicaRESUMEN
INTRODUCTION: Fascioliasis and dicrocoeliasis are the most frequent zoonotic diseases with increasing human health problems in different parts of Iran. Two species, Fasciola hepatica (F. hepatica) and Fasciola gigantica (F. gigantica), are spread in the country. Molecular approaches have a decisive role in identifying both the species. The aim of this study was to detect Fasciola spp. and Dicrocoelium spp. by amplifying the ITS-2 and 28S rDNA gene sequence. METHODS: Overall, 30 infected liver samples were collected from the livestock of Qazvin, Iran. The adult flukes were collected from different livestock. DNA extraction and PCR amplification of ribosomal RNA gene region (ITS2) and 28S rDNA gene fragment were conducted and a phylogenetic tree was constructed. RESULT: All the isolates obtained from the cattle (No: 7) and 82.6% (No: 19) of sheep isolates were infected with F. hepatica species, whereas 17.4% (No: 4) of sheep isolates were infected with F. gigantica. It was also shown that F. hepatica was the predominant species of Fasciola present in the region. All the specimens were infected with Dicrocoelium dendriticum (D. dendriticum). CONCLUSION: Both the species of Fasciola were found in Qazvin. D. dendriticum was the sole infecting species of the Dicrocoelium genus in the livestock of the city of Qazvin. Further research studies are needed to determine the intermediate host of the parasites in the region.
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ADN Espaciador Ribosómico/genética , Dicroceliasis/parasitología , Dicrocoelium/clasificación , Fasciola/clasificación , Fascioliasis/parasitología , Ganado/parasitología , ARN Ribosómico 28S/genética , Animales , Bovinos , ADN Protozoario/genética , ADN Ribosómico/genética , Dicrocoelium/genética , Dicrocoelium/aislamiento & purificación , Fasciola/genética , Fasciola/aislamiento & purificación , Humanos , Irán , Hígado/parasitología , Tipificación de Secuencias Multilocus , Filogenia , Ovinos , Zoonosis/parasitologíaRESUMEN
Fasciola gigantica and hybrid Fasciola are distributed throughout Asia. Herein, we investigated the species of the Fasciola fluke distributed in three hotspots of fascioliasis in Cambodia. A total of 92 flukes collected from 21 slaughtered cattle from Kandal (44), Battambang (41), and Kratie (7) Provinces were identified as F. gigantica using multiplex PCR for a nuclear phosphoenolpyruvate carboxykinase (PEPCK) gene. The overall prevalence of F. gigantica infestation was 7.14% (21/294). Phylogenetic as well as population genetics analyses were performed using the mitochondrial NADH dehydrogenase subunit 1 (ND1). The 19 ND1 haplotypes were identified from Cambodian F. gigantica (haplotype diversity, 0.83). All of the haplotypes were classified into F. gigantica haplogroup C, which includes ND1 haplotypes detected from Thailand, Vietnam, Indonesia, Myanmar, and China. Among haplogroup C, novel and unique haplotypes of Cambodia were found in the Battambang and Kandal Provinces, and the nucleotide diversity of the Cambodian population (0.00532) was the highest. Pairwise fixation indices among the F. gigantica populations from these countries indicated that the Cambodian and Thailand populations were related to each other. The highest genetic diversity in the Cambodian population suggests that F. gigantica in Cambodia may be the ancestor of the populations in Southeast Asian countries. Most likely, livestock movement, including Zebu cattle, played an important role in the transmission of F. gigantica. In this study, the hybrid Fasciola flukes that are commonly found in neighboring countries, were not found in Cambodia. Further comprehensive investigations of Fasciola prevalence should be conducted by analyzing a wider range of hosts throughout Cambodia to reach a more solid conclusion about the absence of hybrid flukes.
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Distribución Animal , Enfermedades de los Bovinos/parasitología , Fasciola/clasificación , Fasciola/genética , Fascioliasis/veterinaria , Variación Genética , Animales , Asia Sudoriental/epidemiología , Cambodia , Bovinos , Enfermedades de los Bovinos/epidemiología , Fascioliasis/epidemiología , Fascioliasis/parasitología , Haplotipos , NADH Deshidrogenasa/genética , Fosfotransferasas/genética , PrevalenciaRESUMEN
Fasciola spp. are the causative agents of fascioliasis in humans and livestock. Before the development of control and management measures, the geographical distribution of the species and patterns of infection must be considered. Because of difficulties in the phenotypic differentiation and morphometric classification of Fasciola spp., DNA molecular markers have become more useful for fluke differentiation and description of phylogenetic patterns. This study aimed to differentiate and describe the phylogenetic background of Fasciola spp. isolated from cattle slaughtered at three abattoirs in the Mpumalanga and KwaZulu-Natal provinces of South Africa. The cytochrome c oxidase I (COI) - FHCO1 (forward: 5'-TTGGTTTTTTGGGCATCCT-3') and FHCO1 (reverse: 5' -AGGCCACCACCAAATAAAAGA3') - marker was sequenced from 55 Fasciola flukes that were collected from abattoirs in catchment areas of the KwaZulu-Natal and Mpumalanga provinces. Fasciola hepatica was demonstrated to have 100% prevalence in KwaZulu-Natal and Mpumalanga (highveld), respectively, and 76% prevalence in the lowveld (Belfast area) of Mpumalanga. Two animals from the Belfast metapopulation were co-infected with both Fasciola gigantica and F. hepatica. DNA sequence analysis of all the isolates demonstrated a sequence conservation of 0.472, nucleotide diversity of 0.082 and Tajima's D of -1.100; however, it was not statistically significant (p > 0.05). Twenty-two haplotypes were identified, with 18 novel haplotypes being unique to the isolates from South Africa. Within the study samples, 12 haplotypes were isolated to a few individuals, with a haplotype diversity of 0.8957 indicating high genetic diversity. Principal coordinate analysis supported the clustering and distribution of the haplotypes, with 11.38% of the variation being attributed to coordinate 2 and 55.52% to coordinate 1. The distribution of Fasciola spp. has been demonstrated to be related to the distribution of the freshwater intermediate host snails, Lymnaea spp., as well as the relative altitude of the localities in South Africa. Information provided by this study serves as preliminary evidence for further studies on the mapping of the distribution of F. gigantica and F. hepatica in South Africa, which is key in designing control programmes for fascioliasis in humans and livestock.
Asunto(s)
Enfermedades de los Bovinos/epidemiología , Complejo IV de Transporte de Electrones/genética , Fasciola/aislamiento & purificación , Fascioliasis/veterinaria , Microbiología de Alimentos , Mataderos , Animales , Bovinos , Enfermedades de los Bovinos/parasitología , ADN de Helmintos/análisis , Fasciola/clasificación , Fasciola/genética , Fascioliasis/epidemiología , Marcadores Genéticos , Filogenia , Sudáfrica/epidemiologíaRESUMEN
A systematic review was conducted focusing on the distribution of Fasciola species and their snail intermediate hosts (IHs) in East and Southern Africa. The reviewed literature showed that both Fasciola hepatica and F. gigantica are present in East and Southern Africa, and infect a wide range of domestic and wild ruminants. Fasciola gigantica was reported in six East African and five Southern African countries, where Radix natalensis (found in low altitudes) was reported to be the main IH. Fasciola hepatica was reported in Tanzania and Ethiopia (East Africa), and in South Africa and Zimbabwe (Southern Africa), where Galba truncatula (found in high altitudes) was documented as the IH in all countries except in Zimbabwe. Both Fasciola species were documented in Tanzania, Ethiopia, Zimbabwe and South Africa. An overlap of the two was observed in areas with an intermediate altitude in Ethiopia and South Africa, where Pseudosuccinea columella was widespread and assumed to transmit both species. Pseudosuccinea columella has been reported in South Africa and Namibia, and proven to transmit F. gigantica in South Africa; its role in Namibia in the transmission of Fasciola species has not been reported. Other lymnaeid species such as R. rubiginosa were reported in South Africa, and R. auricularia in South Africa and Botswana; their role in the transmission of Fasciola species has not been proven. Future studies should aim to determine the role of P. columella in the geographical spread of the two species in East and Southern African countries.
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Fasciola/clasificación , Fascioliasis/veterinaria , Caracoles/parasitología , Vertebrados/parasitología , Animales , Vectores de Enfermedades , Fasciola/fisiología , Femenino , Geografía , Interacciones Huésped-Parásitos , Masculino , Sudáfrica , Especificidad de la EspecieRESUMEN
Fasciola jacksoni is a significant contributor to the health and mortality of Asian elephants, particularly those in Sri Lanka. Despite the impact of fascioliasis on elephant populations, it is a neglected veterinary disease with limited taxonomic understanding. Molecular characterization and phylogenetic analysis of F. jacksoni were carried out to evaluate its suggested basal position in the Fasciolidae. Adult worms were collected during post-mortem of elephants, and eggs were collected from living elephants in National parks across Sri Lanka. Using the mitochondrial genes nicotinamide dehydrogenase subunit 1 (nad1) and cytochrome oxidase subunit 1 (cox1), and a partial 28S ribosomal DNA (28S rDNA), DNA sequences were generated from the F. jacksoni adult and egg material. Maximum likelihood (ML) phylogenetic analyses did not resolve F. jacksoni to be basal to the Fasciolidae. Furthermore, the ML analyses showed that the genus Fasciola was not monophyletic and that F. jacksoni was a sister species to the deer liver fluke Fascioloides magna. A clear framework is required to determine the taxonomic status of F. jacksoni and this current study provides the first detailed application of molecular techniques from multiple hosts across Sri Lanka with the production of reference DNA sequences for this important parasite.
Asunto(s)
Elefantes/parasitología , Fasciola/clasificación , Fasciola/genética , Fascioliasis/veterinaria , Filogenia , Animales , Análisis por Conglomerados , ADN de Helmintos/química , ADN de Helmintos/genética , ADN Mitocondrial/química , ADN Mitocondrial/genética , ADN Ribosómico/química , ADN Ribosómico/genética , Complejo IV de Transporte de Electrones/genética , Fasciola/aislamiento & purificación , Fascioliasis/parasitología , NADH Deshidrogenasa/genética , ARN Ribosómico 28S/genética , Análisis de Secuencia de ADN , Sri LankaRESUMEN
Fasciolosis, a food- and waterborne infection caused by the trematodes Fasciola hepatica and F. gigantica, is recognized by WHO as a neglected zoonotic disease. Whereas F. hepatica is distributed worldwide in cooler climates, F. gigantica occurs mainly in the tropics of Africa and Asia. The southern Caucasus, with Armenia, is one of the most northern regions where both species occur and may produce hybrids. In this study, livestock in central Armenia was surveyed for fasciolosis, the causative species were determined and the genetic diversity of both species was estimated. Total prevalence in sheep (1794), cattle (324) and goats (9) was 21.2%, 15.7% and 44.4%, respectively. After morphological identification and sequencing of a mitochondrial (nad1) and a nuclear marker gene (28S rRNA), 62 collected specimens were allocated to F. hepatica (n = 55) and F. gigantica (n = 7). Intraspecific diversity was evaluated for the complete nad1 gene, resulting in 29 haplotypes of F. hepatica and six haplotypes of F. gigantica. Diversity was higher among F. gigantica than F. hepatica in the Armenian sample set, a difference that was confirmed analyzing available sequences for both species worldwide. Maximum genetic distance between haplotypes in global networks was 49 nucleotide steps for F. gigantica compared to 15 for F. hepatica. In the available sample sets, F. hepatica showed higher diversity in western Asia and the Middle East compared to Europe and eastern Asia, while for F. gigantica loosely structured clusters comprising mainly western/southern Asian and African haplotypes could be identified. A distinct clade comprising haplotypes from Zambia was basal in the phylogenetic tree. Biogeographical implications of these data are discussed.
Asunto(s)
Fasciola/genética , Fascioliasis/veterinaria , Variación Genética , Ganado/parasitología , Filogenia , Animales , Armenia/epidemiología , Asia/epidemiología , Bovinos , ADN de Helmintos , Europa (Continente)/epidemiología , Fasciola/clasificación , Fasciola hepatica/genética , Fascioliasis/epidemiología , Fascioliasis/parasitología , Marcadores Genéticos , Cabras , Haplotipos , Medio Oriente/epidemiología , NADH Deshidrogenasa/genética , Filogeografía , Prevalencia , ARN Ribosómico 28S/genética , Ovinos , Zambia/epidemiologíaRESUMEN
Fasciola hepatica and F. gigantica are ruminant liver flukes that are found worldwide. They can occur sympatrically and hybridize, consequently producing an intermediate form (Fasciola sp.), especially in Africa and Asia. The bridge intron (TkBridgeInt) and intron 4 of domain 2 (TkD2Int4) of the taurocyamine kinase gene were cloned and sequenced from 14 F. hepatica, 18 F. gigantica, and 12 Fasciola sp. These intron regions could be used to differentiate F. hepatica from F. gigantica and revealed genetic hybridization and DNA recombination between F. hepatica and F. gigantica in Fasciola sp. from Japan and Bangladesh. Two specimens of F. hepatica from Ecuador revealed cryptic diversity. A combination of three haplotypes observed in a Fasciola sp. specimen from Japan suggested triploidy. Thus, the intron region of the taurocyamine kinase gene is a potential marker for detecting the hybrid status of the Fasciola flukes.
Asunto(s)
ADN de Helmintos/genética , Fasciola/genética , Hibridación Genética , Recombinación Genética , Animales , Bangladesh , Búfalos/parasitología , Bovinos/parasitología , Clonación Molecular , ADN de Helmintos/química , Ecuador , Fasciola/clasificación , Fasciola/aislamiento & purificación , Variación Genética , Haplotipos , Intrones , Japón , Análisis de Secuencia de ADNRESUMEN
Nuclear gene markers, phosphoenolpyruvate carboxykinase (pepck) and DNA polymerase delta (pold), have been developed for precise discrimination of Fasciola flukes instead of internal transcribed spacer 1. In this study, these two genes of 730 Fasciola flukes from eight Asian countries were analyzed. The results were compared with their mitochondrial NADH dehydrogenase subunit 1 (nad1) lineages for obtaining a definitive evidence of the hybrid origin of aspermic Fasciola flukes. All the flukes categorized into the aspermic nad1 lineages possessed both the fragment patterns of F. hepatica and F. gigantica (mixed types) in pepck and/or pold. These findings provide clear evidence for the hybrid origin of aspermic Fasciola lineages and suggest that "aspermic Fasciola flukes" should hereafter be called "hybrid Fasciola flukes".
Asunto(s)
ADN Polimerasa III/genética , Fasciola/genética , Fosfoenolpiruvato Carboxiquinasa (ATP)/genética , Animales , Asia , Quimera , Fasciola/clasificación , NADH Deshidrogenasa/genéticaRESUMEN
Fasciola gigantica is considered the major pathogen causing fasciolosis in Africa; however, molecular characterization of this fluke has not been adequately elucidated. It is important to scientifically elucidate the dispersal history of F. gigantica by analyzing its genetic diversity. Fasciola flukes from Nigeria were analyzed using nuclear and mitochondrial DNA markers. A total of 172 Fasciola flukes collected from cattle were identified as F. gigantica because they displayed the F. gigantica fragment pattern in multiplex PCR for the nuclear marker, phosphoenolpyruvate carboxykinase (pepck). In total, 70 haplotypes were detected from Nigerian F. gigantica on the basis of the concatenated sequence of mitochondrial NADH dehydrogenase subunit 1 (nad1) and cytochrome c oxidase 1 (cox1). The index of neutrality (Fu's Fs) suggests rapid expansion of the Nigerian F. gigantica population. Although four haplogroups, Nigeria 1A, 1B, 2A, and 2B, were detected from Nigerian F. gigantica, a climate-specific genetic structure was not observed among F. gigantica populations from three agro-climatic regions (Sahel, Savannah, and Forest). This is probably because of the frequent transportation of livestock from one part of the country to the other. Nigeria 1A and 1B had close relationships with the Egyptian population of F. gigantica, whereas Nigeria 2A and 2B were comparatively related to the Zambian population. No haplotype was shared among the three countries, and it therefore is difficult to estimate the dispersal route of F. gigantica within the African continent.
Asunto(s)
Enfermedades de los Bovinos/parasitología , Fasciola/clasificación , Fasciola/genética , Fascioliasis/veterinaria , África/epidemiología , Animales , Bovinos/parasitología , Enfermedades de los Bovinos/epidemiología , ADN de Helmintos/genética , ADN Mitocondrial/genética , ADN Espaciador Ribosómico/genética , Egipto/epidemiología , Fasciola/aislamiento & purificación , Fascioliasis/epidemiología , Fascioliasis/parasitología , Variación Genética , Haplotipos , Nigeria/epidemiología , Fosfoenolpiruvato Carboxiquinasa (ATP)/genética , Filogenia , Reacción en Cadena de la Polimerasa , Polimorfismo de Longitud del Fragmento de Restricción , Análisis de Secuencia de ADNRESUMEN
BACKGROUND: In Egypt, liver flukes, Fasciola spp. (Digenea: Fasciolidae), have a serious impact on the farming industry and public health. Both Fasciola hepatica and Fasciola gigantica are known to occur in cattle, providing the opportunity for genetic recombination. Little is known on the identity and genetic variability of Fasciola populations in sheep. METHODS: This study was performed to determine the prevalence of liver flukes in sheep in Menofia Province as a representative area of the delta region in Egypt, as measured by postmortem examination of slaughtered animals at three abattoirs. The identity and genetic variability of Fasciola spp. in slaughtered animals were determined by PCR-sequence analysis of the nuclear ribosomal internal transcribed spacer 1 (ITS1) and the mitochondrial NADH dehydrogenase subunit 1 (nad1) genes. RESULTS: Physical inspection of the liver indicated that 302 of 2058 (14.7%) slaughtered sheep were infected with Fasciola spp. Sequence analysis of the ITS1 and nad1 genes of liver flukes from 17 animals revealed that 11 animals were infected with F. hepatica, four with F. gigantica, and two with both species. Seventy eight of 103 flukes genetically characterized from these animals were F. hepatica, 23 were F. gigantica, and two had ITS1 sequences identical to F. hepatica but nad1 sequences identical to F. gigantica. nad1 sequences of Egyptian isolates of F. gigantica showed pronounced differences from those in the GenBank database. Egyptian F. gigantica haplotypes formed haplogroup D, which clustered in a sister clade with haplogroups A, B and C circulating in Asia, indicating the existence of geographic isolation in the species. CONCLUSIONS: Both F. hepatica and F. gigantica are prevalent in sheep in Egypt and an introgressed form of the two occurs as the result of genetic recombination. In addition, a geographically isolated F. gigantica population is present in the country. The importance of these observations in epidemiology of fascioliasis needs to be examined in future studies.