Fasciola Gigantica Research Articles

MORPHOLOGICAL AND BIOINFORMATICS STUDY FOR RADIX AURICULARIA SNAILS IN FRESHWATER IN BASRAH PROVINCE, IRAQ

Radix auricularia (Lymnaea auricularia) snails are the main intermediate host of Fasciola gigantica in the middle and South of Iraq. Recently, some studies suggest these snails are not present in Iraq, in particular Basrah province. Thus, the current study was aimed to use the bioinformatics analysis in the determination of Radix sp. in Basrah and also aimed to identify morphological characteristics of this species in the AL-Sewaib river, Basrah. Four hundred R. auricularia snails were collected from the AL-Sewaib river during May 2018 to March 2019. The results of the bioinformatics analysis showed according to the mitochondrial cytochrome c oxidase subunit I, the presence of R. auricularia snails in Basrah province and these snails are more closely related to Iranian R. auricularia snails than those for European counties and Russia. This study also displayed the existence of the polymorphism phenomenon in R. auricularia snails collected from the AL-Sewaib river which there were six different forms (A, B, C, D, E and F). These forms had variations in the colour and form of shells along with a variation in the number of eggs (30-56) per an egg mass. In addition to this, these R. auricularia forms had variations in their distributions. Form E had the highest spread percentage (33%) followed by 17, 15, 14, 11 and 10% for forms B, D, C, A and F, respectively. These forms had also variations in their morphological characteristics. The largest form was the form D whereas the smallest form was the form B but other forms A, C, E and F were similar and had moderate sizes.Thus, the current study concludes the existence of R. auricularaia snails in Basrah as a representative city for the southern part of Iraq and also confirmed the presence of the polymorphism phenomenon in the members of R. auricularaia obtained from the AL-Sewaib river.

Morphological and molecular characterization of Fasciola hepatica and Fasciola gigantica phenotypes from co-endemic localities in Mpumalanga and KwaZulu-Natal provinces of South Africa

Fasciolosis is a food- and water-borne disease caused by digenean trematode species, Fasciola hepatica and F. gigantica. They are widely distributed and infect a wide range of definitive hosts, causing enormous economic loss due to reduced productivity in domestic ruminants. The two species have been previously reported to be co-endemic in KwaZulu-Natal and Mpumalanga provinces of South Africa. Hybridization between the two species has been reported elsewhere. Despite the overlap of the two species in two provinces, there has been no attempt to determine the presence of the intermediate forms or hybrids. Therefore, this study aimed at morphological and molecular characterization of Fasciola spp. collected from cattle slaughtered at abattoirs located in the two provinces of South Africa, where two species are endemic. A total of 71 fluke specimens were collected cattle from abattoirs in Enhlazeni and Nelspruit in Mpumalanga province and Pietermaritzburg in KwaZulu-Natal province of South Africa, and Zimbabwe (Bulawayo abattoir). Fasciola gigantica specimen collected from Zimbabwe where it has been confirmed as the only species occurring and this was used as control in the morphological and molecular assessment of the collected specimens. Of the 71 specimens collected, 37 were classified as F. hepatica, 12 as F. gigantica and 22 as Fasciola spp using morphological characters. Of these species, 11 of 37F. hepatica and 6 of 22 Fasciola sp were found to be aspermic or having very scanty sperm. Fifteen flukes which were spermatic were all identified morphologically as F. gigantica whilst 5 flukes which were aspermic were identified morphologically as F. hepatica. Molecular analysis of the same 15 spermatic specimens confirmed the presence of F. hepatica (n=9) and F. gigantica (n=6) using the CO1 marker and as F. hepatica (n=4), F. gigantica (n=7) and Fasciola sp. (n=1) for the same specimens using the ITS-1/5.8S/ITS-2 marker. The remaining 4 aspermic flukes (one did not resolve) were all identified morphologically as F. hepatica and molecular analysis confirmed them as F. hepatica (n=4) by both CO1 and ITS-1/5.8S/ITS-2. Phylogenetic analysis based on both CO1 and ITS-1/5.8S/ITS-2 showed that F. hepatica species formed a moderately supported monophyletic clade with F. gigantica. Their ancestral history was further confirmed by haplotype network, which formed novel haplotypes that corresponded with the structure of the phylogenetic tree. Results from this study showed that morphological characters alone have limitations in identifying F. hepatica and F. gigantica in areas where the two species occur, although both methods confirmed the presence of F. gigantica occurring in Zimbabwe, F. hepatica in KwaZulu-Natal, and both species occurring in Mpumalanga province. Therefore, the use of morphological techniques, complemented by molecular techniques are recommended, especially in endemic areas where the two species are co-endemic.

Molecular characterization and phylogenetic analyses of Fasciola gigantica of buffaloes and goats in Punjab, Pakistan.

Fasciola gigantica is considered to be a major pathogen causing fasciolosis in the Indian subcontinent, resulting in production losses of millions of dollars in the livestock industry. Understading the dispersal origin and the patterns of spread of F. gigantica is important. A total of 53 Fasciola flukes collected from buffaloes and goats in Punjab, Pakistan between 2017 and 2018 were identified as F. gigantica based on the multiplex PCR for the phosphoenolpyruvate carboxykinase (pepck) and the PCR-restriction fragment length polymorphism (RFLP) for DNA polymerase delta (pold). A significant genetic difference between F. gigantica from buffaloes and goats was indicated by the genetic analyses of mitochondrial markers, NADH dehydrogenase subunit 1 (nad1) and cytochrome C oxidase subunit 1 (cox1). Phylogenetic analysis of the seventeen nad1 haplotypes of F. gigantica from Pakistan with those in neighbouring countries of the Indian subcontinent revealed that all the haplotypes identified in Pakistan were clustered in haplogroup A. fasciola gigantica with the eight haplotypes might be expanded in Pakistan from Indian origin, along with the migration of the domestic animals, since they were related to Indian haplotypes. In contrast, the remaining nine haplotypes were not shared with any neighbouring countries, suggesting independent origin, probably from neighbouring Middle East countries. However, cautious interpretation is required due to the very limited samples size of this study. Our study provides a proof of concept for a method that could be used to investigate the epidemiology of F. gigantica.

Differential expression of microRNAs and tRNA fragments mediate the adaptation of the liver fluke Fasciola gigantica to its intermediate snail and definitive mammalian hosts

The tropical liver fluke Fasciola gigantica affects livestock and humans in many Asian countries, large parts of Africa, and parts of Europe. Despite the public health and economic impacts of F. gigantica, understanding of F. gigantica biology and how the complex lifecycle of this liver fluke is transcriptionally regulated remain unknown. Here, we tested the hypothesis that the regulatory small non-coding RNAs (sncRNAs), microRNAs (miRNAs) and tRNA-derived fragments (tRFs) play roles in the adaptation of F. gigantica to its intermediate and definitive hosts. We sequenced sncRNAs of eight lifecycle stages of F. gigantica. In total, 56 miRNAs from 33 conserved families and four Fasciola-specific miRNAs were identified. Expression analysis of miRNAs suggested clear stage-related patterns. By leveraging the existing transcriptomic data, we predicted a miRNA-based regulation of metabolism, transport, growth and developmental processes. Also, by comparing miRNA complement of F. gigantica with that of Fasciola hepatica, we detected a high level of conservation and identified differences in some miRNAs, which can be used to distinguish the two species. Moreover, we found that tRFs at each lifecycle stage were predominantly derived by tRNA-Lys and tRNA-Gly at 5′ half sites, but relatively high expression was related to the buffalo-infecting stages. Taken together, we provided a comprehensive overview of the dynamic transcriptional changes of small RNAs that occur during the developmental stages of F. gigantica. This global analysis of F. gigantica lifecycle stages revealed new roles of miRNAs and tRFs in parasite development and will facilitate future research into understanding of fasciolosis pathobiology.

Evaluation of flow through assay as a point-of-care test for detection of antibodies against Fasciola gigantica in sheep

A novel flow through assay (FTA) was evaluated for detection of antibodies against Fasciola gigantica in ovines, employing excretory and secretory antigens (ESA) and somatic antigens (SOA) prepared from adult F. gigantica and serum samples from sheep (n=240) that were positive and negative for F. gigantica and positive for amphistomes. Of the sheep sera sampled, 38.75% and 35.42% were found to have antibodies against F. gigantica with ESA and SOA respectively. An ELISA protocol was used as confirmatory test. In comparison with necropsy inspection and faecal examination as standard, the ESA-FTA showed higher specificity (95.2%), sensitivity (100.0%) and accuracy (96.4%) compared to that of SOA-FTA (90.9, 100 and 93.0 %) with a K value of 0.98 indicating perfect agreement. Screening of serum samples by ESA-ELISA also revealed highest specificity (93.8%), sensitivity (100.0%) and accuracy (95.2%) compared to that of SOA-ELISA (88.2, 100 and 90.9%). The FTA results were comparable with faecal examination and ELISA and can be used as a point-of-care test for the detection of fasciolosis in sheep.

Фасциолез в субъектах Северного Кавказа как вероятная биологическая, эпидемиологическая и эпизоотическая угроза населению и животноводству

When analyzing statistical materials from 32 polyclinics in 15 cities of the North Caucasus (Grozny, Gudermes, Argun, Magas, Nazran, Vladikavkaz, Beslan, Ardon, Karachaevsk, Cherkessk , Nalchik, Nartkala, Baksan, Prokhladny and Mayskiy) in 18,0 thousand samples of feces, the indices of occurrence per 100 thousand of population and abundance of F. gigantica eggs were 0, which indicates an epidemic the well-being of the region in relation to zoonosis, thanks to the program organization of health education of the population. At the same time, for fascioliasis (Fasciola gigantica) of sheep, cattle and buffaloes, all 5 Republics of the North Caucasus belong to the disadvantaged regions of southern Russia with a critical risk index for the biosafety of livestock industries in all climatic zones of the subject due to the weak and half implementation of antiepizootic measures. In 5 Republics of the North Caucasus in 2011–2019. The indices of the incidence and abundance of fascioliasis in sheep, cattle and buffaloes increased by 2–3 times, which indicates the threat of invasion to the efficiency of livestock subsectors in the region. Our data indicate a high level of sanitary contamination of the soils of near-village pastures in the plain and foothill zones of all subjects of the North Caucasus with invasive elements of the causative agent of giant fascioliasis and their absence in the soils of distant pastures. In 100% of soil samples from rural pastures in the plain and foothill zone, invasive eggs of Fasciola gigantica were identified, which, according to forecasts, may lead to the spread of parthenitis among intermediate hosts, followed by the spread of invasion among the population of 5 regions, such as the Chechen and Ingush Republics, RNOAlania, Kabardino-Balkarian and Karachay-Cherkess Republics.

Fasciolosis Infection Level of Various Breed Cattle in Batu and Pujon District, East Java, Indonesia

Fasciolosis is a disease caused by Fasciola gigantica (Cobbold, 1855) or Fasciola hepatica (Linnaeus, 1758) that infected ruminants and caused liver damaged and loss of productivity. To control Fasciola disease in cattle, it is necessary to consider differences in the prevalence or number of cases in various cattle breed. This research was conducted in two slaughterhouses in Batu and Pujon District, Malang Regency, East Java, Indonesia involving 50 livestock in each slaughterhouse, either beef or dairy cattle. Observation was done for 2 mo by performing liver necropsy, observing the presence of fibrosis and calcification of the bile duct, counting the number of worms and weighing the removed liver tissue. Result of this research the number of fasciolosis in Ongole and Limousin cattle (30.95 % and 35.94 %) was fewer compared to Simmental and Frisian Holstein (58.33 %). Average number of worms head–1 in each cattle breed; Limousin, Simmental, Ongole, and FH were (105, 49, 27 and 129) worms. Meanwhile, fibrosis score in each group were 1.48, 1.89, 1.33 and 1.75 respectively. It is suspected that differences in cattle breed had correlation with susceptibility towards infection. It is indicated that Limousin cattle are more tolerant against liver worm infection.

Prevalence and factors influencing gastrointestinal parasitic infections in sheep in Bangladesh.

Gastrointestinal (GI) parasites are major contributors to decrease productivity in livestock over the world. A cross-sectional study was conducted in different areas of Bangladesh to determine the prevalence of GI parasitic infections and their association with the biotic and abiotic factors in sheep. A total of 572 faecal samples were collected from the selected areas of Bangladesh and microscopic examination was performed for the identification of parasites using flotation and sedimentation technique. Out of 572, 441 animals were found infected with one or more species of GI parasites with an overall prevalence of 77.1%. Nine types of parasites from four different classes were detected namely Strongyles (42.1%), Strongyloides sp. (27.1%) and Trichuris sp. (1.0%), Moniezia sp. (2.4%), Paramphistomum cervi (32.5%), Fasciola gigantica (6.1%) and Schistosoma sp. (3.5%), coccidia (16.6%) and Balantidium coli (7.9%). Nematodes infections (56.8%) were significantly highest among trematodes (37.9%), protozoa (24.4%) and cestode (2.4%). In the present study, all the biotic factors including sex, age, physiological condition of female and body condition score (BCS) of animals were insignificantly (p>0.05) associated with the prevalence of GI parasitic infection in sheep but among the abiotic factors, muddy housing of animals, rainy season, having no knowledge about GI parasites and illiteracy of farmers were significantly (p<0.05) associated with the GI parasitic infections. This epidemiological investigation will assist to build a suitable control program against GI parasites in sheep and thus, help to prevent production loss and increase livelihood of small holder farmers.

Comments on "Genetic characterization and phylogenetic analysis of Fasciola species based on ITS2 gene sequence, with first molecular evidence of intermediate Fasciola from water buffaloes in Aswan, Egypt".

Dear Editor-in-Chief, In Annals of Parasitology 2021, 67(1), 55-65, a paper entitled "Genetic characterization and phylogenetic analysis of Fasciola species based on ITS2 gene sequence, with first molecular evidence of intermediate Fasciola from water buffaloes in Aswan, Egypt" was published with great interest [1]. After reading the article carefully and critically, we think some points should be noted. Fasciola species are meiotically functional diploid, can produce sperm and temporarily and store in the seminal vesicles. This type is named spermic fluke [2]. On the other hand, intermediate Fasciola with morphological characteristics intermediates between F. hepatica and F. gigantica with no sperm or aspermic and no sperm in seminal vesicles. However, this is also seen in older flukes [3-5]. It seems that morphological studies based on spermatogenesis ability were necessary for this study. Also, this parasite's anthelmintic resistance is due to aspects of biology, and population structure depends on genetic diversity [6]. We question whether there are any documents about and sequences of mitochondrial markers as COX (Cytochrome Oxidase) and NAD (Nicotinamide Adenine Dinucleotide) to analyze intraspecific phylogenetic relationship in addition to nuclear gene? In Table 3, the pairwise distances between three groups of Fasciola spp. from different livestock animals were low, ranging from 0.004 to 0.01 with an overall mean of 0.008. Genetic diversity is described as a tendency of genetic characteristics to vary and serves as a way for the population to adapt to changing hosts and environments [7]. The nature of the nuclear gene (ITS) is instability. It is better to use mitochondrial sequence data to compare diversity. Also, genetic discrimination grade from infra population to meta population is annotated by Fst value ranging; 0 to 1. Fst values between 0-0.05 indicated a low genetic differentiation population [8]. It seems that by calculating Fst and showing the gene migration based on mitochondrial sequences data of specimens, this study's species population will be obtained. Also, Tajima's D and Fu's F in all loci populations based on GenBank data may show the Fasciola haplotypes' population proximity. Here we recommend, that Omar et al. [1] studies that molecular phylogeny with mitochondrial DNA efectively used for appropriate diferentiation of haplotypes and spermatogenic ability by carmen allium staining helps them find the physiological aspects. Of course, more prominent populations are needed to find intermediate types. [1] Omar M.A, Elmajdoub L.O., Ali A.O., Ibrahim D.A., Sorour S.S., Al-Wabel M.A., Suresh M., Metwally A.M. 2021. Genetic characterization and phylogenetic analysis of Fasciola species based on ITS2 gene sequence, with first molecular evidence of intermediate Fasciola from water buffaloes in Aswan, Egypt. Annals of Parasitology 67: 55-65. doi:10.17420/ap6701.312 [2] Sanderson A. 1953. Maturation and probable gynogenesis in the liver fluke, Fasciola hepatica L. Nature 172: 110-112. doi:10.1038/172110a0 [3] Hayashi K., Ichikawa-Seki M., Mohanta U.K., Singh T.S., Shoriki T., Sugiyama H., Itagaki T. 2015. Molecular phylogenetic analysis of Fasciola flukes from eastern India. Parasitology International 64: 334-338. https://doi.org/10.1016/j.parint.2015.04.004 [4] Ichikawa-Seki M., Tokashiki M., Opara M.N., Iroh G., Hayashi K., Kumar U.M., Itagaki T. 2017. Molecular characterization and phylogenetic analysis of Fasciola gigantica from Nigeria. Parasitology International 66: 893-897. doi:10.1016/j.parint.2016.10.010 [5] Rouhani S., Raeghi S., Mirahmadi H., Fasihi Harandi M., Haghighi A., Spotin A. 2017. Identification of Fasciola spp. in the east of Iran, based on the spermatogenesis and nuclear ribosomal DNA (ITS1) and mitochondrial (ND1) genes. Archives of Clinical Infectious Diseases 12:e57283. doi:10.5812/archcid.57283 [6] Hodgkinson J., Cwiklinski K., Beesley N., Paterson S., Williams D., Devaney E. 2013. Identification of putative markers of triclabendazole resistance by a genome-wide analysis of genetically recombinant Fasciola hepatica. Parasitology 140: 1523. doi:10.1017/S0031182013000528 [7] Bozorgomid A., Rouhani S., Harandi M.F., Ichikawa- Seki M., Raeghi S. 2020. Genetic diversity and distribution of Fasciola hepatica haplotypes in Iran: molecular and phylogenetic studies. Veterinary Parasitology: Regional Studies and Reports 19: 00359. [8] Rouhani S., Raeghi S., Spotin A. 2017. Spermatogenic and phylo-molecular characterizations of isolated Fasciola spp. from cattle, North West Iran. Pakistan Journal of Biological Sciences 20: 204-209.

Prevalence of Fasciolosis in Commercial Cattle Farm of Tilottama Municipality, Rupandehi, Nepal

Fasciolosis is a common parasitic disease affecting cattle and other ruminants, caused by Fasciola hepatica and Fasciola gigantica. The disease is widely distributed and can cause extensive economic losses to the farmers and the national economy. A cross sectional study was conducted to determine the prevalence of fasciolosis in commercial cattle farms of Tilottama Municipality, Rupandehi district, Nepal. A total of 270 fresh faecal samples were collected purposively from the study area with different ages, sex, stages, and breeds for faecal examination (sedimentation method) to diagnose eggs of Fasciola in the collected samples. The obtained data were coded and analysed using Microsoft Excel 2016. The overall prevalence of fasciolosis in cattle was found 15.56%. Age and sex wise prevalence was found statistically significant (P&lt;0.05) while stage and breed wise prevalence was insignificant (P&gt;0.05). Therefore, fasciolosis is prevalent moderately among cattle in Tillottoma municipality, which necessitates the study of detailed epidemiology of the disease and effective control strategies against it in order to prevent huge economic losses.

Identification of Adult Fasciola spp. Using Matrix-Assisted Laser/Desorption Ionization Time-of-Flight (MALDI-TOF) Mass Spectrometry

Fascioliasis is a neglected trematode infection caused by Fasciola gigantica and Fasciola hepatica. Routine diagnosis of fascioliasis relies on macroscopic identification of adult worms in liver tissue of slaughtered animals, and microscopic detection of eggs in fecal samples of animals and humans. However, the diagnostic accuracy of morphological techniques and stool microscopy is low. Molecular diagnostics (e.g., polymerase chain reaction (PCR)) are more reliable, but these techniques are not routinely available in clinical microbiology laboratories. Matrix-assisted laser/desorption ionization time-of-flight (MALDI-TOF) mass spectrometry (MS) is a widely-used technique for identification of bacteria and fungi; yet, standardized protocols and databases for parasite detection need to be developed. The purpose of this study was to develop and validate an in-house database for Fasciola species-specific identification. To achieve this goal, the posterior parts of seven adult F. gigantica and one adult F. hepatica were processed and subjected to MALDI-TOF MS to create main spectra profiles (MSPs). Repeatability and reproducibility tests were performed to develop the database. A principal component analysis revealed significant differences between the spectra of F. gigantica and F. hepatica. Subsequently, 78 Fasciola samples were analyzed by MALDI-TOF MS using the previously developed database, out of which 98.7% (n = 74) and 100% (n = 3) were correctly identified as F. gigantica and F. hepatica, respectively. Log score values ranged between 1.73 and 2.23, thus indicating a reliable identification. We conclude that MALDI-TOF MS can provide species-specific identification of medically relevant liver flukes.

Perubahan Patologi Sistem Hepatobiliari Sapi Bali Terinfeksi Fasciola gigantica

Liver fluke infections is an important parasitic disease that common attack cattle, buffalo and others ruminant. The disease in the tropics is caused by Fasciola gigantica trematodes. Fasciolosis found in Bali cattle besides causing high economic loss also has the opportunity infect humans. This study aims to determine the pathology changes of Bali cattle hepatobiliary system infected by F. gigantica. In January 2019 were examined a hundred of liver tissue Bali cattle. Based on the sex as many as 75 bali cattle are cows and 25 are bulls. On macroscopic examination was found fifteen bali cattles in gallbladder containing of F. gigantica. In positive Fasciolosis the surface of liver look uneven with enlargement of lymph nodes. The tissue of Bali cattle liver infected with F. gigantica is cut into 1x1x1 cm and put in a pot that has been filled with 10% neutral buffered formaline. The fixed tissue then processed in a tissue processor and stained with hematoxylin-eosin (HE). Microscopic examination was found bile ducts proliferation and blocked of the lumen accompanied by cholangio cirrhosis. Found longitudinal pieces of Fasciola gigantica liver fluke surrounded by fibrous connective tissue and multifocal coagulative necrosis with fibroblast proliferation. In some location the infiltration of neutrophil cells around the bile ducts can also be seen. Congestion, bleeding, and neutrophil infiltration are also seen in areas of necrosis. In the bile duct wall bleeding, adenomatus hyperplasia, infiltration of mononuclear cells with mild intensity and foci of calcification are found. It can be concluded that there was a change with severe intensity accompanied by chronic inflammation in the hepatobiliary system of Bali cattle with Fasciolosis.

Incidence of gastro intestinal parasites in cattle of fringe villages of chakrashila wild life sanctuary in B.T.A.D. (Assam) in post monsoon period

Chakrashila Wildlife Sanctuary is a wildlife sanctuary falling under the two districts of Lower Assam under BTAD (Bodoland Territorial Area Districts) of Dhubri and Kokrajhar and covers approximately 45.568 kms2. The humid climatic condition of Assam is very conducive for the growth and multiplication of parasites and cause clinical and subclinical parasitism. Subclinical infections are responsible for high morbidity and mortality in young animals and enormous production losses in adults. Different species of gastrointestinal parasites including zoonotic parasites of public health importance have been reported from cattle of this region. The monsoon season is the ideal time for the mass propagation of parasitic load in environment. The average overall prevalence rate occurrence of gastrointestinal parasites in cattle was found to be 64.45% in these villages wherein the main parasitic load was of Fasciola gigantica, Toxocara vitulorum, Eimeria sp., Amphistome sp., and Buxtonella sp. in both Choraikhola and Harinaguri villages wherein there was a gradual decrease in parasitic load from the month of October towards December-2018. Coccidiosis due to Eimeria sp. from Choraikhola and Harinaguri were found to be the predominating disease with an overall prevalence of (44.45%) and (62.23%) respectively, which is in accordance with the earlier findings of coccidiosis in buffalo calves in winter months in India.

Prevalence and Phylogenetic analysis of Fasciola species in Upper Egypt Based on Ribosomal ITS-2 gene Sequencing

Fascioliasis is an important parasitic zoonosis with an estimated annual economic loss at more than US$3 billion in animals and infection risk in more than 180 million people worldwide. Given the economic and public health significance of this trematode, a periodical evaluation of its prevalence and the associated risk factors as well as the evolutionary relatedness is needed. Here, 1560 cattle and 1630 goat livers were examined for the presence of adult Fasciola among animals slaughtered in slaughterhouses of at Assiut and Sohag Governorates, Upper Egypt. Molecular characterization and phylogenetic relationships were determined based on the second internal transcribed spacer (ITS-2) of nuclear ribosomal DNA (rDNA). The overall prevalence of fasciolosis was based on morphological identification of the recovered Fasciola adult worms and the prevalence was 5.1% and 3.5% among cattle and goats, respectively. Though the age, gender, and housing system significantly affected the disease in cattle, none of these variables had a significant impact on goat fasciolosis. Nonetheless, the sampled goats were found at increased risk of being infected with Fasciola hepatica as the age increases and when the winter season emerges. Phylogenetic analyses showed that both Fasciola gigantica and Fasciola hepatica sequenced in this study had a common ancestor and they fall in one clade with several previously reported Egyptian and worldwide fasciolae from animals and humans. This study demonstrates that the disease is currently circulating among animals in Assiut and Sohag Governorates that warrants the urgent need for application of the appropriate control measures as well as evaluating the possible ongoing zoonotic burden in people residing such areas.

Dysregulation of hepatic microRNA expression in C57BL/6 mice affected by excretory-secretory products of Fasciola gigantica.

The excretory-secretory products released by the liver fluke Fasciola gigantica (FgESPs) play important roles in regulating the host immune response during the infection. Identification of hepatic miRNAs altered by FgESPs may improve our understanding of the pathogenesis of F. gigantica infection. In this study, we investigated the alterations in the hepatic microRNAs (miRNAs) in mice treated with FgESPs using high-throughput small RNA (sRNA) sequencing and bioinformatics analysis. The expression of seven miRNAs was confirmed by quantitative stem-loop reverse transcription quantitative PCR (qRT-PCR). A total of 1,313 miRNAs were identified in the liver of mice, and the differentially expressed (DE) miRNAs varied across the time lapsed post exposure to FgESPs. We identified 67, 154 and 53 dysregulated miRNAs at 1, 4 and 12 weeks post-exposure, respectively. 5 miRNAs (miR-126a-3p, miR-150-5p, miR-155-5p, miR-181a-5p and miR-362-3p) were commonly dysregulated at the three time points. We also found that most of the DE miRNAs were induced by FgESPs in the mouse liver after 4 weeks of exposure. These were subjected to Gene Ontology (GO) enrichment analysis, which showed that the predicted targets of the hepatic DE miRNAs of mice 4 weeks of FgESPs injection were enriched in GO terms, including cell membrane, ion binding, cellular communication, organelle and DNA damage. KEGG analysis indicated that the predicted targets of the most downregulated miRNAs were involved in 15 neural activity-related pathways, 6 digestion-related pathways, 20 immune response-related pathways and 17 cancer-related pathways. These data provide new insights into how FgESPs can dysregulate hepatic miRNAs, which play important roles in modulating several aspects of F. gigantica pathogenesis.

A PIONEER USAGE OF GOLD NANOPARTICLES IN SANDWICH ELISA BASED ON CIRCULATING HYDATID ANTIGEN AS A TOOL FOR IMPROVEMENT OF SERODIAGNOSTIC TESTING

Human cystic echinococcosis is a zoonosis occurring due to dogs handling and exposure toEchinococcus granulosus ova in their stools being an accidental intermediate host. Serologyremains the only sure and important tool to diagnose this disease. We aimed to improve thedetection of circulating antigen by sandwich ELISA through its binding to gold nanoparticleand proving the role of minute nanoparticles in serodiagnosis. Gold nanoparticles (AuNPs)were employed for the capture of the antigens of protoscolices (pAg) in the patient’s serum bysandwich ELISA. Cross-reactivity for antigens from Hymenolepis nana, Entrobius vermicularis,and Fasciola gigantica was ruled out by using anti-protoscolices polyclonal IgG antibodies(ppAb). Antigen of sonicated protoscolices which were removed from camel lungcysts, was purified and injected to a New-zealand white rabbit giving ppAb which then loadedon AuNPs being used as a diagnostic indicator for circulating pAg by both sandwich ELISAand nanogold sandwich ELISA techniques. Nanogold sandwich ELISA was able to give positiveresults with 96.3% of hydatid patients and 5% of non-hydatid patients while sandwichELISA showed 81.4% and 20% positive cases of the same groups respectively. The sensitivityand specificity of nanogold sandwich ELISA were 96.3% & 95% against 81.5% & 80% forthose of sandwich ELISA respectively. The conjugation of AuNPs to anti-Echinococcus IgGantibodies seems to be beneficial to increase PPV, NPV, and efficiency and reduce the overallcost of the assay as less antibody was needed.

IMMUNOLOCALIZATION OF EXCRETORY/SECRETORY ANTIGENS EXPRESSED ON DIFFERENT LIFE CYCLE STAGES AND IN DIFFERENT ORGANS OF FASCIOLA GIGANTICA INFECTED CATTLE

In this study, polyclonal antibodies (pAb) were prepared against Fasciola gigantica excretory/secretory (E/S) products and purified to demonstrate the highly reactive epitopes on the differentlife cycle stages of F. gigantica and on blood cells and different organs of naturally infectedcattle, as well as to screen internal organs for localization of E/S antigens to find out a suitablemethod for diagnosing the disease as early as possible. Using anti-F. gigantica E/S IgG pAb,immunostaining of different life cycle stages of F. gigantica detected strong positive reactions inall stages except eggs. The reactions were along tegumental surface, gut region of cercariae,miracidiae, metacercariae and tail of cercariae. Besides, the tegument, spine surface, muscularisand gut region of the adults were also positive. Immunoperoxidase localization of E/S productsby anti-F. gigantica E/S IgG pAb showed that E/S antigens were located in many organs ofnaturally infected cattle. Examination of peripheral blood mononuclear cells (PBMNCs)represented a high positivity in the phagosomes and activated monocytes as well as in cytoplasmof lymphocytes. Compared to sections of normal uninfected hosts, liver sections of infectedbovine hosts showed collapsed hepatocytes, thickened portal tract and a highly positive reactionin the cytoplasm of the liver cells. Target epitopes were located inside the splenic cells, mainlylymphocytes. In the gall bladder, positive peroxidase reactions were found in the mucosa andmuscularis layers, and in the epithelial cells lining mucosal glands. A marked positivity was inother organs of the naturally infected animals, as in the epithelial layer lining the glomeruli andtubules of the kidney, in heart cardiac muscle fibres and in cells lining the alveoli of the lungs.

Novel expression of Cathepsin B Fasciola gigantica sequence for detection of human fascioliasis in Egypt

Novel expression of Cathepsin B Fasciola gigantica sequence for detection of human fascioliasis in Egypt

Prevalence of Fasciola gigantica in Sheep and Goats and Associated Risk Factors in Zaria, Kaduna State, Nigeria

Fasciolosis is a parasitic disease affecting ruminants. The aim of the study was to determine the prevalence of Fasciola gigantica in sheep and goats in Zaria, Kaduna State. A total of 400 faecal samples were collected over a period of three months (July to September, 2018) and examined using the formal ether sedimentation technique. The overall intensity of the infection was one plus(1+) indicating low infection. The prevalence of F. gigantica in the small ruminants examined was 52(13.0%) with sheep having a higher prevalence of 34(17.0%) than in goats 18(9.0%). Higher prevalence of F.gigantica was also observed in adult sheep 28(22.2%) and goats 15(20.0%) than in young ones of sheep 6(8.1%) and goats 3(2.4%).Consequently, higher prevalence of F. gigantica was observed in female sheep and goats with respective prevalence rates of 23(24.2%) and 17(12.1%) than in male sheep and goats with rates of 11(10.5%) and 1(1.7%) respectively. The prevalence of F. gigantica was 8(21.6%), 23(17.0%) and 3(10.7%) for Uda, Yankasa, and Balami breeds of sheep respectively, and 17(10.4%) and 1(2.7%) for Kano Brown and Crosses of goats respectively. Higher prevalence of F. gigantica was observed in ruminants on non-settled farms 37(18.5%) than those on settled farms15(7.5%).The prevalence of F. gigantica was significantly (p&lt;0.05) higher in animals with poor body condition 18(34.0%) than good body condition 1(1.8%) animals. This study has established the presence of F. gigantica in faecal sample of sheep and goats in farms within Zaria, therefore appropriate use of anthelmintics and awareness of the public on the prevention and control measures should be ensured.

Méthodes comparées de dépistage de &lt;em&gt; Fasciola gigantica &lt;/em&gt; chez les bovins dans un abattoir de l’Ouest Cameroun

Une enquête a été réalisée sur 330 bovins à l’abattoir de Bangangté (région Ouest, Cameroun) pour estimer la prévalence de la fasciolose bovine. Trois méthodes de dépistage ont été comparées : a) l’inspection vétérinaire post mortem, suivie de la dissection des principaux canaux biliaires destinée à mettre en évidence les adultes de Fasciola gigantica ; b) la coproscopie, utilisant la technique de sédimentation ; et c) la recherche des oeufs dans la bile. Cette dernière méthode, considérée comme la technique de référence, a permis d’identifier 110 bovins (prévalence de 33 %) infestés. Des douves adultes ont été retrouvées dans le foie de 24 d’entre eux (prévalence de 7 %). Quant à la coproscopie, elle était positive chez 20 animaux (prévalence de 6 %). Tous les bovins positifs à la coproscopie ou lors de l’examen du foie avaient des oeufs de F. gigantica dans les échantillons de bile. En revanche, parmi les 306 bovins chez lesquels aucune douve adulte n’a été retrouvée dans le foie, 10 avaient des oeufs de F. gigantica dans leurs fèces et 86 éliminaient des oeufs dans la bile. Comparées à la technique de référence, les sensibilités de la coproscopie et de la recherche des douves adultes dans les foies ont été respectivement de 18 % (20 coproscopies vs 110 animaux avec oeufs dans la bile) et de 22 % (24 foies positifs vs 110). Ces résultats montrent que la fasciolose bovine est fréquente chez les animaux abattus à Bangangté, mais que l’examen du foie ou la coproscopie ne permettent pas d’identifier tous les animaux infestés.