Fasciola Gigantica Research Articles

Prevalence of fasciolosis emphasis on age, origin, body condition and post mortem by geographic information systems on sacrificial examination in Malang District – East Java

Infection with Fasciola gigantica, a liver trematode, is frequently reported in Malang district. The study was aimed to determine the prevalence of fascioliasis in Malang District, East Java during Eid Qurban 2019. Data were collected from 68 cutting points in 9 sub-districts. A total of 265 cows, 895 goats and 112 sheep than analyzed based on questioner emphasis on age, origin, body condition, and postmortem abnormalities. Overall the highest prevalence of fasciolosis was 57% cattle and 18% goat in Gondanglegi sub-district and 100% sheep in Wajak sub-district. A total of 86% of fasciolosis in cattle have good body conditions came from Malang District. There was also 82% in goat and 78% in sheep infected came from Malang District too. Morphological and postmortem examination findings that 53% of adult cattle (2-3 years), adult goat and sheep (1-1,5 years) were 78% and 89% respectively have abnormalities in the liver. This data showed adult livestock more resistant to fasciolosis infection. From that data above that further surveys should be carried out to understanding on the epidemiology of fascioliasis in the study area and classified risk factors affecting the infection.

Biochemical and immunological investigation of fascioliasis in cattle in Egypt.

Background and Aim:Fasciola hepatica and Fasciola gigantica are two commonly reported liver flukes that cause fascioliasis in ruminants. Among the members of the genus Fasciola, F. hepatica was identified in the study area. Fascioliasis is a major disease that affects the production of livestock by causing liver damage. F. hepatica has developed advanced mechanisms to trick, elude, and alter the host immune response, similar to an extrinsic stressor. These mechanisms consequently affect the animals’ physiological and metabolic functions in vivo and postmortem changes, which have significant influences on animal welfare and meat quality development. Therefore, this study aimed to determine the current prevalence of cattle fascioliasis at abattoirs in El-Kharga city, New Valley Governorate, Egypt, and to investigate the changes in serum biochemical and immunological parameters and oxidative stress factors due to Fasciola spp. infection in terms of meat quality and immune response.Materials and Methods:A total of 226 cattle were inspected for the presence of Fasciola spp. The liver of each cattle was examined by making several incisions for detecting adult Fasciola spp. in El- Kharga. The blood samples were collected to analyze the changes in serum biochemical and immunological parameters and oxidative stress factors.Results:Of the 226 cattle, 38 (16.81%) were positive for F. hepatica at the postmortem examination. Cattle infected with F. hepatica had highly elevated serum alanine aminotransferase, aspartate aminotransferase, glutamate dehydrogenase, γ-glutamyl transferase, urea, and creatinine levels. Immunological cytokine profiles showed significantly increased serum interleukin (IL)-4, IL-10, and transforming growth factor-beta levels and a significantly decreased interferon-γ level. Furthermore, oxidative stress profiles showed significantly increased serum malondialdehyde and nitric oxide levels and significantly decreased total antioxidant capacity and reduced glutathione level.Conclusion:This study demonstrated that F. hepatica infection alone is an oxidative stress factor that affects slaughtered animals, leading to biochemical and metabolic alterations in the early postmortem period.

Helminth infections of great concern among cattle in Nigeria: Insight to its prevalence, species diversity, patterns of infections and risk factors.

Background and Aim:Helminth infections are one of the greatest causes of productive and reproductive loss in animals and man, and in some cases, it results in heavy mortalities. This study was conducted to determine the prevalence, species diversity, patterns of infections and risk factors associated with helminth infections of cattle in Ilorin, Nigeria.Materials and Methods:A total of 478 fecal samples were collected from abattoirs and cattle farms over a year period (March, 2018-February, 2019). Fecal samples were visually examined then observed using simple flotation and formalin-ethyl acetate sedimentation techniques. Eggs and worms were identified according to standard procedures. The packed cell volume was determined using the hematocrit centrifugation technique.Results:A total of 79.92% of the cattle examined were found positive with one or more helminth species. Eighteen helminth species (cutting across all classes of helminths) were detected, with Haemonchus contortus (60.46%), Trichostrongylus spp. (46.44%), Ostertagia ostertagi (42.05%), Bunostomum phlebotomum (28.87%), Cooperia spp. (24.27%), Oesophagostomum radiatum (21.97%), Strongyloides papillosus (12.13%), and Fasciola gigantica (10.67%) been the most prevalent. Helminth infection was detected all through the year with the least prevalence recorded in February (55.00%). About 61% of the examined cattle harbored double/multiple helminth species. There was a significant difference between breed, sex, physiological status, and season with the prevalence rate of helminth infections (p<0.05).Conclusion:Our investigation demonstrated high prevalence and wide diversity of helminth species, which suggests that helminth infections are of great concern among cattle in Ilorin and Nigeria in general. There is a need for a radical veterinary intervention to curb the menace so as to have an economically robust cattle industry in Nigeria.

Genotyping study of Fasciola gigantica isolated from cattle in Aqrah city, Iraq

This study was conducted to investigate the 18S rRNA gene of Fasciola gigantica obtained from the liver of cattle live in Aqrah city, Iraq. Fifty-nine Fasciola flukes were collected through routine investigation from livers of naturally infected local cattle (42 cows), from May to August 2017, at the central slaughterhouse of Aqrah city, Kurdistan region of Iraq, the flukes were washed by PBS and then fixed in ethanol. Genomic DNA was extracted, and a 560 bp fragment was amplified by PCR, subsequent by sequencing of PCR products. A remarkable result of this project was the deposition of our gene isolate in GenBank (Accession No. MG786553). However, it was confirmed by the sequence results that isolate species was F. gigantica, and interestingly our samples sequences have alignment match of 100% with many international isolates, without genetic mutations or variations. It is concluded that molecular study could be utilized for both diagnosis and differential diagnosis of parasites with huge precise. Also, an 18S rRNA gene is a perfect fragment for molecular study and phylogenetic analysis of F. gigantica, also our samples have 100% alignment match with universal isolates.

Molecular phylogenetic and genetic variability of Fasciola gigantica in Kermanshah province, western Iran with an overview to understand haplotypes distribution in Asia and Africa.

Over the last decade, diagnostic tools to detect and differentiate Fasciola species have improved, but our understanding of the distribution of haplotypes and population structure of this parasite is less clear. This study was designed to survey this gap in the F. gigantica epidemiology in Kermanshah province, western Iran from 2015 to 2017. Sixty-eight Fasciola isolates were collected from slaughterhouses from this province. We evaluated the PCR-RFLP assay of the ITS1 genes for the identification of Fasciola species using the RsaI enzyme. After Fasciola species identification, the partial sequence of mitochondrial NADH dehydrogenase subunit 1 (ND1) gene of F. gigantica was used for subsequent construction of the phylogenetic tree and network analysis. Based on the PCR-PRFLP profile, one (6.25%) of sheep isolates and 19 (39.60%) of cattle isolates were detected as F. gigantica, whereas 93.75% of sheep isolates, 60.40% of cattle isolates and all of the goat isolates were F. hepatica. In the 20 analyzed flukes, five ND1 haplotypes were detected. Statistically significant genetic differentiation was demonstrated between the Iran population and all the other populations. Evidence is presented for the existence of two well-separated populations: African and West Asian gigantica flukes and East Asian gigantica flukes. Genetic relationships among haplotypes were associated with geographical divisions. Also, our results have heightened our knowledge about the genetic diversity of F. gigantic, providing the first evidence for the existence of two well-separated populations of this parasite.

Anthelmintic Activity of Curcuma Aeruginosa Roxb Extract on Fasciola gigantica in Vitro

Fasciola gigantica is a parasite that causes a disruption of the metabolism of fats, proteins, and carbohydrates, which interferes growth and causes death. Curcuma aeruginosa Roxb extract is one of the medicinal plants which has been used to treat several diseases. The aim of this study was to determine the effect of methanolic extract of C. aeruginosa Roxb on F. gigantica, including mortality time and histopathological changes that occurred after treatment. This study used a completely randomized design with five replications. The flukes were soaked in three different extract concentrations: 10% (T1), 25% (T2), and 50% (T3) (w/v). Phosphate buffered saline (PBS) solution was used as a negative control (C1) and albendazole as the positive control (C2). The mortality time of F. gigantica in each group was calculated, and the dead flukes were prepared for histopathological study. The data were analyzed by Analysis of variance. The results showed that C. aeruginosa extracts at the concentration of 10, 25 and 50% caused the death of the flukes within 75, 57 and 48 minutes, respectively., Histopathological observations showed that the extract caused breakage of tegument which is an important organ in the respiratory process and nutrient absorption. This study concluded that C. aeruginosa extract exhibited anthelmintic activity towards F. gigantica in vitro.

The Anthelmintics Activity of Ethanol Extract of African Leaf (Vernonia amygdalina) on Mortality of Fasciola gigantica in Vitro

The Aims of this study is to know the anthelmintics activity of ethanol extract of african leaf (Vernonia amygdalina) on mortality of Fasciola gigantica in Vitro. Method that used in the research was a completely randomized design. There were five treatments and each treatment was done in four replications. This research used ten Fasciola gigantica in each treatment for all replications. The observation and recording of dead Fasciola gigantica was done at 2, 4, 6, 8, and 10 hours. Fasciola gigantica were declared dead if there was no movement when disturbed by anatomy tweezers and when dipped in slightly warm water (500C). The obtained data was analyzed using ANOVA and continued with Duncan Multiple Range Test. The result of this research show that ethanol of extract of African Leaf (Vernonia amygdalina) has activity anthelmintic on mortality of Fasciola gigantica in Vitro. Ethanol extract african leaf (Vernonia amygdalina) with concentration of 1.4% is the optimal concentration of anthelmintics. This higher the concentration of the extract, the higher the property of anthelmintics.

Genetic diversity and distribution of Fasciola hepatica haplotypes in Iran: Molecular and phylogenetic studies.

Fasciolosis is a zoonotic disease caused by Fasciola hepatica and Fasciola gigantica. Over the last decade, diagnostic tools to detect and differentiate Fasciola species have improved, but our knowledge of the distribution of haplotypes and gene flow of this parasite is not comprehensive yet. The purpose of this study was to investigate this gap in the epidemiology of F. hepatica in different provinces of Iran between 2015 and 2017. Isolated Fasciola were collected from abattoirs in 9 provinces. The partial sequence of mitochondrial NADH dehydrogenase subunit 1 (ND1) gene was used for the identification and molecular analysis of F. hepatica isolates. The amplified PCR products were purified and subjected to direct sequencing for subsequent construction of phylogenetic tree and network analysis. In the 130 subjects analyzed, 37 ND1 haplotypes were detected. This is the first study in Iran which investigates F. hepatica population and its genetic structure, based on mitochondrial ND1 marker in different geographical regions of Iran.

Newly excysted juveniles (NEJs) of Fasciola gigantica induce mice liver fibrosis and M2 macrophage-like phenotype in vivo

Liver flukes of animals are parasitic flatworms of major socioeconomic importance in many countries. Particularly, Fasciola gigantica is a leading cause of production losses to the livestock (mainly sheep and cattle) and meat industries due to clinical disease, reduced weight gain and milk production, and deaths. Immune responses induced by helminth have been extensively studied, but there is limited information on this aspect by F. gigantica, especially on macrophages induced with this parasite. Studies have shown that host immune responses induced by parasitic infection is greatly correlated with the macrophage polarization axis. In the present study, we used the murine model of F. gigantica to explore the interaction of host and F. gigantica. We found F. gigantica NEJs promoted pathology and fibrosis of mice liver, and the enlargement of mice spleen. We also showed that macrophages were recruited to mice peritoneal cavity at 5 days post infection. By evaluating the expression of genetic markers of M2 macrophages such as Arg-1, Ym1 and RELMɑ, and genetic marker of M1 macrophages iNOS, we showed that M2 macrophages were induced by F. gigantica. M2 macrophages are central to the immune response during helminth infection, and our findings in this study provided insight into the immune interaction between F. gigantica and host.

Utilization of Curcuma Molases Block to Increase The Performance of Goats that Infected Gastrointestinal Worm at Wonorejo, Wates District, Kediri Regency

This was a Community Service activity that aims to share knowladge about utilization of Curcuma Molasses Block (CMB) to increase performance of goats that infected gastrointestinal (GI) parasites to The Goat Breeder Group at Wonorejo, Wates District, Kediri Regency. The activities include: counseling about raising goat management, presentasion on worm diseases that infect goats and CMB as anthelmentic, workshop and practice to make CMB, detection egg worm in goat`s feces, treatment on infected goats and filling out questionnaires evaluating the performance of goat before and after giving CMB. The result show that all of feces samples were positive contain egg worms of Haemonchus contortus, Chabertina ovina, Trichuris globulosa, Strongyloides papillosus, Fasciola gigantica, Trichostrongylus spp., Toxocara sp., and Oesohagustomum columbianum even also protozoans Eimeria sp., and Blastocystis sp. Administration CMB increase performance of worm-infected goats which indicated by increase in appetite, soft and shiny hair and the body more well-fed.

Generation of oxidative stress and induction of apoptotic like events in curcumin and thymoquinone treated adult Fasciola gigantica worms

Fasciolosis is a neglected tropical disease caused by the liver fluke Fasciola gigantica. The absence of successful vaccine and emerging resistance in flukes against the drug of choice, triclabendazole, has necessitated the search for alternatives including phyto-therapeutic approaches. Curcumin and thymoquinone, the active ingredients of Curcuma longa and Nigella sativa plants respectively, were first screened for their binding affinity with Glutathione-S-transferase (GST) molecule through in silico molecular docking followed by in vitro treatment of worms with varying concentrations of the test compounds.The in silico molecular docking of curcumin and thymoquinone with sigma GST revealed strong hydrogen bonding as well as hydrophobic interactions with high fitness scores but showing inter-specific differences. The in vitro treatment of F. gigantica worms with both curcumin and thymoquinone resulted in a significant increase in the generation of reactive oxygen species (ROS) whereas the level of reduced glutathione, a primary redox regulator, was found to be significantly decreased (p < 0.05). The two compounds not only inhibited the GST activity, which is an important detoxification enzyme and also a key drug/vaccine target for the control of fasciolosis but also significantly inhibited the activity of antioxidant enzymes glutathione peroxidase and glutathione reductase that are vital in maintenance of redox homeostasis. The immunohistochemistry performed using anti sigma GST polyclonal antibodies revealed that both the compounds used in the present study significantly reduced immunofluorescence in the vitellaria, developing eggs present in the ovary and the intestinal caecae indicating inhibition of GST enzyme in these regions of the worms. Further, following treatment with curcumin and thymoquinone, chromatin condensation and DNA fragmentation was also observed in F. gigantica worms.In conclusion, both curcumin and thymoquinone generated oxidative stress in the worms by production of ROS and significantly inhibiting their antioxidant and detoxification ability. The oxidative stress along with induction of apoptotic like events would compromise the survival ability of worms within the host. However, further studies are required to establish their anthelmintic potential alone and in combination with the commonly used anthelmintic drugs under in vivo conditions.

Detections of gastrointestinal parasites, including Giardia intestinalis and Cryptosporidium spp., in cattle of Banten province, Indonesia.

Gastrointestinal parasites can induce low productivity in livestock by causing acute or chronic enteritis. Veterinarians make great efforts to design rational and effective hygienic protocols for both the prevention and treatment of diarrhea. Although prevalences can vary depending on the examined areas or the ages of the hosts, and the methods used for detections, it is helpful to accumulate data across many areas to evaluate parasitic distribution. A coprological survey in cattle was conducted in Tangerang, Banten Province of Indonesia, in order to determine the prevalence of the parasites, including those of diarrhea-associated diseases. Furthermore, the risk of transmission of Giardia intestinalis and Cryptosporidium spp. to human was genetically analyzed. Gastrointestinal parasites were detected in 87 of 109 cattle samples, including 85 carrying Eimeria spp., 36 carrying Fasciola gigantica, 35 carrying Strongyloides spp., 33 carrying Paramphistomum spp., and 15 carrying Capillaria spp. Giardia intestinalis and Cryptosporidium spp., parasites with zoonotic potential, were detected in 9 and 1 cattle samples, respectively. Molecular analyses identified the G. intestinalis isolate as a member of Assemblage E, which has been recently detected in humans in another country. These results may be helpful in understanding the hygienic risk affecting the livestock productivity and zoonotic potential of cattle in Indonesia.

Prevalence of Fascioliasis and Dicrocoeliasis in Cattle Slaughtered in Some Abattoirs in Akure Metropolis, Ondo State, Nigeria

The study was carried out in March-May 2019 to investigate the prevalence of fascioliasis and dicrocoeliasis in cattle slaughtered in parts of Akure abattoir, Ondo State, Nigeria. Two hundred (200) bile samples from cattle were collected from both male and female cattle. The parasitological analysis of the bile sample was carried out by sedimentation technique. Prevalence of Fasciola gigantica was (50.5%) for Dicrocoelium hospes was 16.0% and double infections accounted for (5.5%). Prevalence of Fasciola gigantica infections was high in female cattle (57.7%) compared to their male counterparts (38.9%). For Dicrocoelium hospes, the infection was higher in male (18.2%) than in the female (14.6%) while the prevalence of double infections was higher in males (7.8%) than in the females (4.07). The high infection rate for Fasciola gigantica (84.0%) and Dicrocoelium hospes (62.0%) was found to be high at Busa abattoir. Double infection was observed to be high at Oke-Odu abattoir (10%) than in other abattoirs while no infection was observed at Bola meat abattoir. Overall age prevalence of fascioliasis and dicrocoeliasis (82.8%) was within the age bracket of 9-10yrs while the least infection (54.2%) fall within the age bracket of 7-8yrs. Prevalence of fascioliasis and dicrocoeliasis in Sannia breed was 90.2% followed by the Ambala breed (84.0%) while the least was 31.2% in the White Fulani breed. This study which revealed a high prevalence of fascioliasis and dicrocoeliasis among cattle slaughtered in the studied abattoir possesses a threat to healthy livestock productions and it is significant public health importance. Adequate measures to improving the veterinary services are needed to put in place to reduce the economic losses accruing to these infections.

Monoclonal antibody against Fasciola gigantica glutathione peroxidase and their immunodiagnosis potential for fasciolosis

Glutathione peroxidases (GPx), major antioxidant enzymes, secreted by Fasciola spp., are important for the parasite evasion and protection against the host’s immune responses. In the present study, a monoclonal antibody (MoAb) against recombinant F. gigantica glutathione peroxidase (rFgGPx) was produced by hybridoma technique using spleen cells from BALB/c mice immunized with rFgGPx. This MoAb (named 7B8) is IgG1 with κ light chains, and it reacted specifically with rFgGPx at a molecular weight 19 kDa as shown by immunoblotting, and reacted with the native FgGPx in the extracts of whole body (WB), metacercariae, newly excysted juveniles (NEJs), 4 week-old juveniles and adult F. gigantica as shown by indirect ELISA. It did not cross react with antigens in WB fractions from other adult trematodes, including Fischoederius cobboldi, Paramphistomum cervi, Setaria labiato-papillosa, Eurytrema pancreaticum, Gastrothylax crumenifer and Gigantocotyle explanatum. By immunolocalization, MoAb against rFgGPx reacted with the native protein in the tegument, vitelline cells, and eggs of adult F. gigantica. In addition, the sera from mice experimentally infected with F. gigantica were tested positive by this indirect sandwich ELISA. This result indicated that FgGPx is an abundantly expressed parasite protein that is secreted into the tegumental antigens (TA), therefore, FgGPx and its MoAb may be used for immunodiagnosis of both early and late fasciolosis gigantica in animals and humans.

Triclabendazole in the treatment of human fascioliasis: a review.

Fascioliasis occurs on all inhabited continents. It is caused by Fasciola hepatica and Fasciola gigantica, trematode parasites with complex life cycles, and primarily affects domestic livestock. Humans become infected after ingestion of contaminated food (typically wild aquatic vegetables) or water. Fascioliasis may be difficult to diagnose as many symptoms are non-specific (e.g. fever, abdominal pain and anorexia). Treatment options are limited, with older effective therapies such as emetine and bithionol no longer used due to safety issues and unavailability, and most common anthelminthics having poor efficacy. Clinical trials conducted over a 25-year period, together with numerous case reports, demonstrated that triclabendazole has high efficacy in the treatment of human fascioliasis in adults and children and in all stages and forms of infection. Triclabendazole was approved for human use in Egypt in 1997 and in France in 2002 and a donation program for the treatment of fascioliasis in endemic countries was subsequently established by the manufacturer and administered by the World Health Organization. Here the published data on triclabendazole in the treatment of human fascioliasis are reviewed, with a focus on more recent data, in light of the 2019 US Food and Drug Administration approval of the drug for use in human infections.

Complex and dynamic transcriptional changes allow the helminth Fasciola gigantica to adjust to its intermediate snail and definitive mammalian hosts

BackgroundThe tropical liver fluke, Fasciola gigantica causes fasciolosis, an important disease of humans and livestock. We characterized dynamic transcriptional changes associated with the development of the parasite in its two hosts, the snail intermediate host and the mammalian definitive host.ResultsDifferential gene transcription analysis revealed 7445 unigenes transcribed by all F. gigantica lifecycle stages, while the majority (n = 50,977) exhibited stage-specific expression. Miracidia that hatch from eggs are highly transcriptionally active, expressing a myriad of genes involved in pheromone activity and metallopeptidase activity, consistent with snail host finding and invasion. Clonal expansion of rediae within the snail correlates with increased expression of genes associated with transcription, translation and repair. All intra-snail stages (miracidia, rediae and cercariae) require abundant cathepsin L peptidases for migration and feeding and, as indicated by their annotation, express genes putatively involved in the manipulation of snail innate immune responses. Cercariae emerge from the snail, settle on vegetation and become encysted metacercariae that are infectious to mammals; these remain metabolically active, transcribing genes involved in regulation of metabolism, synthesis of nucleotides, pH and endopeptidase activity to assure their longevity and survival on pasture. Dramatic growth and development following infection of the mammalian host are associated with high gene transcription of cell motility pathways, and transport and catabolism pathways. The intra-mammalian stages temporally regulate key families of genes including the cathepsin L and B proteases and their trans-activating peptidases, the legumains, during intense feeding and migration through the intestine, liver and bile ducts. While 70% of the F. gigantica transcripts share homology with genes expressed by the temperate liver fluke Fasciola hepatica, gene expression profiles of the most abundantly expressed transcripts within the comparable lifecycle stages implies significant species-specific gene regulation.ConclusionsTranscriptional profiling of the F. gigantica lifecycle identified key metabolic, growth and developmental processes the parasite undergoes as it encounters vastly different environments within two very different hosts. Comparative analysis with F. hepatica provides insight into the similarities and differences of these parasites that diverged > 20 million years ago, crucial for the future development of novel control strategies against both species.

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

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. 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. 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). 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.

Prevalence and risk factors associated with Fasciola gigantica and Paramphistomum cervi infection through actual retrieval in Philippine Carabaos in Ormoc City Abattoir, Philippines

This study determine the prevalence and risk factors associated with Fasciola and Paramphistomum and the co-infection of both parasites in 246 Philippine carabaos slaughtered in Ormoc City abattoir.Upon postmortem examination, adult parasites were collected from the organ of localization and identified based on morphological characteristics. Intrinsic factors [sex, age, weight, Body Condition Score (BCS)] and origin of the carabaos were recorded. The parasites identified were Fasciola gigantica and Paramphistomum cervi. The study showed a prevalence of 50.81% for F. gigantica, 4.31% for P. cervi and 32.11% for coinfection. was more prevalent in female carabaos (57.80%), carabaos more than three years of age (58.41%), &gt;325kg bodyweight (60.36%) and higher than three body condition score (59.09%). Paramphistomum was common in female carabaos (51.37%), carabaos less than or equal to three years of age (46.82%), ≤325kg bodyweight (52.59%) and less than or equal to three (≤3) body condition score (49.64%). Carabaos originating from Leyte have a higher infection rate to Fasciola (53.30%) and Paramphistomum (46.70%) compared to carabaos , supplied by other provinces. Logistic regression modeling indicates that male carabaos are a protective factor (OR 0.59) against Paramphistomum infection while carabaos within Leyte have a greater likelihood of being infected with both parasites (OR 4.27 to 4.57). Pearson’s Correlation Coefficient (r-value) showed a strong and positive result (r=0.997**) that the likelihood of Fasciola and Paramphistomum co-exist (co-infection) together is high. A high prevalence rate of fluke infection has been recorded in carabaos admitted for slaughter implying, that most of the backyard carabao raisers are not practicing sound deworming, pasture and grazing management and proper deworming protocol. To prevent and control flukes and improve production and health of carabaos, the above-mentioned factors must first be considered.

Detecting Fasciola hepatica and Fasciola gigantica microRNAs with loop-mediated isothermal amplification (LAMP).

Fascioliasis is a parasitic infection typically caused by two common parasites of class Trematodo, genus Fasciola, namely Fasciola hepatica and Fasciola gigantica. The widespread of these species in water and food makes fascioliasis become a global zoonotic disease that affects 2.4 million people in more than 75 countries worldwide. Typically, F. hepatica and F. gigantica can be recognized by parasitological techniques to detect Fasciola spp. eggs, immunological techniques to detect worm-specific antibodies, or by molecular techniques such as PCR to detect parasitic genomic DNA. Recently, miRNAs have been raised as a key regulator and potential diagnostic biomarkers of diseases, including parasitic infection. An isothermal PCR called loop-mediated isothermal amplification (LAMP) is rapid, sensitive, and its amplification process is so extensive that making LAMP well-suited for field diagnostics. LAMP reactions for miRNA detection have been introduced and were able to detect the target miRNA amounts in the wide range of 1.0amol to 1.0pmol, exhibiting high selectivity to differentiate one-base between miRNA sequences. Here, we introduced a modified LAMP to detect a species-specific miRNA of F. hepatica and F. gigantica. Our method did not demand an initial heating step and the reactions had a high sensitivity that greater than 1000 times in comparison to that reported in previous studies. Most importantly, the technique could perform well with parasitic miRNA presenting in bovine serum samples without sophisticated equipment required. These results create a promising technique basis for some novel and simple device to diagnose fascioliasis and other parasitic infection diseases at point-of-care.

Genetic relationship of &lt;i&gt;Fasciola gigantica&lt;/i&gt; populations in Africa: Insight from mitochondrial &lt;i&gt;cox 1&lt;/i&gt; genes

The liver fluke Fasciola gigantica is an important livestock parasite causing fascioliasis in Africa. Population genetics analysis of the mitochondrial cox1 genes of F. gigantica from seven African countries revealed two distinct genetic lineages with 41% nucleotide divergence. The first lineage comprised of populations from four countries (Egypt, Mauritania, Nigeria and Zambia) while the second lineage consisted of sequences from Niger, South Africa and Zimbabwe. Mismatch distribution analysis and neutrality tests revealed both F. gigantica lineages are in genetic equilibrium. There was also evidence of gene flow between the populations in the two lineages. The two distinct genetic lineages of F. gigantica identified may have implication for drug resistance in the treatment of fascioliasis.Keywords: Fasciola gigantica; Africa; cox1; genetic diversity; molecular epidemiology.