High-quality reference genome of Fasciola gigantica: Insights into the genomic signatures of transposon-mediated evolution and specific parasitic adaption in tropical regions.

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.

Molecular identification and genetic-polymorphism analysis of Fasciola flukes in Dali Prefecture, Yunnan Province, China.

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.

Fasciolosis in India: An overview.

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.

Molecular characterization revealed Fasciola specimens in Ecuador are all Fasciola hepatica, none at all of Fasciola gigantica or parthenogenic Fasciola species.

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.

Molecular Confirmation of a Fasciola gigantica × Fasciola hepatica Hybrid in a Chadian Bovine.

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.

Genetic diversity and multiplicity of infection in Fasciola gigantica isolates of Pakistani livestock.

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.

Multigene typing and phylogenetic analysis of Fasciola from endemic foci in Iran.

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.

Molecular Characterization of Fasciola and Dicrocoelium Species Isolated from Ruminant Livestock in Qazvin, Iran.

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.

Genetic Characterization of Cambodian Fasciola gigantica and Dispersal Direction of the Species in Asia.

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.

Fasciola species and their vertebrate and snail intermediate hosts in East and Southern Africa: a review.

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.

Phylogenetic analysis of Fasciola spp. isolated from slaughtered cattle in KwaZulu-Natal and Mpumalanga provinces of South Africa based on the cytochrome c oxidase subunit I mitochondrial marker.

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.

Molecular characterization of Fasciola jacksoni from wild elephants (Elephas maximus maximus) of Sri Lanka: a taxonomic evaluation.

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.

Fasciola spp. in Armenia: Genetic diversity in a global context.

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.

Revealing genetic hybridization and DNA recombination of Fasciola hepatica and Fasciola gigantica in nuclear introns of the hybrid Fasciola flukes.

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.

Hybrid origin of Asian aspermic Fasciola flukes is confirmed by analyzing two single-copy genes, pepck and pold.

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".

Molecular characterization and phylogenetic analysis of Fasciola gigantica from Nigeria.

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.

Identity of Fasciola spp. in sheep in Egypt.

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.

Cross-sectional study of Fasciola gigantica and other trematode infections of cattle in Edu Local Government Area, Kwara State, north-central Nigeria.

BACKGROUND: Trematode infections of livestock are of global veterinary and public health importance causing serious economic losses. Majority of data on burden of trematode infections in Nigeria are based on abattoir surveys and there are very few data on herd level risk factors. The present study investigated the prevalence of, and herd level risk factors for, fasciolosis and other trematode infections in cattle in Edu Local Government Area (LGA). METHODS: A cross-sectional survey used two-stage study design to investigate cattle belonging to 65 households. Two questionnaires were administered for household-level and individual cattle-level data. Faecal and blood samples were obtained from the cattle. Logistic regression analyses were performed to determine risk factors for infections. RESULTS: Of 686 faecal samples analysed, 74.9 %, 16.1 %, 7.3 % and 1.2 % were positive for infections with Fasciola gigantica, paramphistomes, Dicrocoelium hospes and Schistosoma bovis respectively. Fasciola gigantica had higher prevalence in adult cattle (77.3 %) than weaners (62.5 %). Majority of co-infections was a combination of F. gigantica with paramphistomes 84/130 (64.6 %). Most (58.9 %) of the cattle belonged to FAMACHA© score 2. The mean packed cell volume (PCV) was 34.4 %. The sensitivity and specificity of FAMACHA© for anaemia (PCV < 24 %) were 18.2 and 96.9 %, respectively. Positive correlation was obtained between faecal egg counts for F. gigantica and paramphistomes (R = 0.15, P = 0.0001). Adult cattle were more likely to be infected with F. gigantica (odds ratio, OR: 1.94; Confidence Interval, CI: 1.19-3.16) than weaners. Cattle belonging to household heads aged between 40-59 years were more likely infected with paramphistomes (OR: 1.95; CI: 1.02-3.74) than those belonging to other age groups. Cattles from herds with size ≥ 100 were more likely infected with D. hospes than those from smaller herds (OR: 6.98; CI: 2.94-16.6). CONCLUSION: This study revealed high prevalence of infection with F. gigantica in Kwara State. The co-infections by F. gigantica and paramphistomes with a positive correlation should be considered during anthelmintic therapy. There is a need to optimise and validate the FAMACHA© for use in cattle based on breeds and variation in colour of ocular mucous membrane. Risk factors identified could assist in tailoring control strategies for various trematode infections to particular groups of farmers and cattle.

Molecular characterization and phylogenetic analysis of Fasciola gigantica from western Java, Indonesia.

Fasciola gigantica and aspermic (hybrid) Fasciola flukes are thought to be distributed in Southeast Asian countries. The objectives of this study were to investigate the distribution of these flukes from unidentified ruminants in western Java, Indonesia, and to determine their distribution history into the area. Sixty Fasciola flukes from western Java were identified as F. gigantica based on the nucleotide sequences of the nuclear phosphoenolpyruvate carboxykinase (pepck) and DNA polymerase delta (pold) genes. The flukes were then analyzed phylogenetically based on the nucleotide sequence of the mitochondrial NADH dehydrogenase subunit 1 (nad1) gene, together with Fasciola flukes from other Asian countries. All but one F. gigantica fluke were classified in F. gigantica haplogroup C, which mainly contains nad1 haplotypes detected in flukes from Thailand, Vietnam, and China. A population genetic analysis suggested that haplogroup C spread from Thailand to the neighboring countries including Indonesia together with domestic ruminants, such as the swamp buffalo, Bubalus bubalis. The swamp buffalo is one of the important definitive hosts of Fasciola flukes in Indonesia, and is considered to have been domesticated in the north of Thailand. The remaining one fluke displayed a novel nad1 haplotype that has never been detected in the reference countries. Therefore, the origin of the fluke could not be established. No hybrid Fasciola flukes were detected in this study, in contrast to neighboring Asian countries.

CIAS detection of Fasciola hepatica/F. gigantica intermediate forms in bovines from Bangladesh.

Fascioliasis is an important food-borne parasitic zoonosis caused by two trematode species, Fasciola hepatica and Fasciola gigantica. The characterisation and differentiation of Fasciola populations is crucial to control the disease, given the different transmission, epidemiology and pathology characteristics of the two species. Lineal biometric features of adult liver flukes infecting livestock have been studied to characterise and discriminate fasciolids from Bangladesh. An accurate analysis was conducted to phenotypically discriminate between fasciolids from naturally infected bovines (cattle, buffaloes) throughout the country. Morphometric analyses were made with a computer image analysis system (CIAS) applied on the basis of standardised measurements and the logistic model of the body growth and development of fasciolids in the different host groups. Since it is the first ever comprehensive study of this kind undertaken in Bangladesh, the results are compared to pure fasciolid populations of F. hepatica from the European Mediterranean area and F. gigantica from Burkina Faso, geographical areas where both species do not co-exist. Principal component analysis showed that the biometric characteristics of fasciolids from Bangladesh are situated between F. hepatica and F. gigantica standard populations, indicating the presence of phenotypes of intermediate forms in Bangladesh. These results are analysed by considering the present emergence of animal fascioliasis, the local lymnaeid fauna, the impact of climate change, and the risk of human infection in the country.