Diagnosis of fasciolosis antibodies in Brazilian cattle through ELISA employing both native and recombinant antigens.

Bovine fasciolosis is a parasitic disease with a global reach. Coprological based on egg detection in fecal samples and liver inspection to evaluate the presence of the parasite is currently the gold standard for diagnosing chronic fasciolosis in cattle. However, these techniques are labor-intensive and ineffective during the acute phase of the disease. Serodiagnosis using native and recombinant antigens has become an interesting alternative in efforts to identify cattle fasciolosis. We evaluated cattle from abattoir (n = 139) and farms (n = 500) through liver inspection and coprological examination, respectively. Our laboratory team optimized and validated enzyme-linked immunosorbent assay tests based on somatic antigen, excretory/secretory proteins, and the recombinant antigen cathepsin L-1 to detect serum antibodies against fasciolosis in cattle. For animals from abattoir, 10 were positive for fasciolosis according to liver inspection. Both FhES and FhrCL-1 presented an area under the receiver operating characteristic (AUROC) curve of 0.80, with a sensitivity of 0.80 (95% CI: 0.46-0.95) and 0.70 (95% CI: 0.38-0.90) and specificity of 0.81 (95% CI: 0.73-0.87) and 0.87 (95% CI: 0.80-0.92), respectively. For those cattle from farms, 28 were positive only for fasciolosis according to coprological examination. In this scenario, FhES gave the best performance, with an AUROC of 0.84, sensitivity of 0.79 (95% CI: 0.60-0.90), and specificity of 0.86 (95% CI: 0.82-0.89). In conclusion, our study highlights the potential of serodiagnosis for accurately screening cattle fasciolosis. The promising sensitivity and specificity values of FhES when compared to liver inspection or coprological examination enhance its importance for cattle fasciolosis diagnosis. IMPORTANCE: The aim of this article was to identify antibodies against fasciolosis in cattle in Brazil. The methodology was reproduced in our laboratory and applied for the first time to the Brazilian cattle herd. The antigens tested can be used as a screening test and thus speed up the diagnosis of bovine fascioliasis.

Validation of the natural sedimentation technique in the diagnosis of chronic fasciolosis.

There are various diagnostic techniques available for chronic fasciolosis in ruminants. However, many of them exhibit low specificity and sensitivity, making them impractical for field use and in low-resource laboratories. The present study evaluates the usefulness of the Natural Sedimentation technique in diagnosing chronic fasciolosis in three domestic species conducted at the Laboratorio de Parasitología y Enfermedades Parasitarias, Facultad de Ciencias Veterinas, Universidad Nacional de Cajamarca. Fecal samples were collected from n = 323 cattle, n = 362 sheep, and n = 231 swine for Fasciola hepatica fecal egg counts. The visualization of adult parasites in animal livers post-mortem was considered the gold standard. Additionally, the sensitivity of the technique was evaluated using five different amounts of feces. In cattle, a sensitivity of 0.93 ± 0.03, specificity of 0.91 ± 0.06, positive predictive value of 0.96 ± 0.03, and negative predictive value of 0.86 ± 0.07 were obtained. In sheep, a sensitivity of 0.79 ± 0.05, specificity of 0.83 ± 0.07, positive predictive value of 0.90 ± 0.04, and negative predictive value of 0.66 ± 0.08 were observed. In swine, a sensitivity of 0.92 ± 0.06, specificity of 1.00 ± 0.00, positive predictive value of 1.00 ± 0.00, and negative predictive value of 0.96 ± 0.03 were found. There was no statistical difference in egg counts when using 1, 2, 3, 4, and 5 g of feces (p = 0.907). Furthermore, 1 to 688 fecal eggs of F. hepatica were counted in 1 g of feces. The Natural Sedimentation technique has both qualitative and quantitative applications with satisfactory results when using 1 g of feces in the diagnosis of chronic fasciolosis in domestic animals. Due to its simplicity, it can be implemented in field conditions and low-resource laboratories.

Fasciola hepatica infection modifies IgG1 specific immune response to foot-and-mouth disease virus induced by vaccination.

Fasciola hepatica, a worldwide distributed helminth, has a robust immunoregulatory effect in the host, increasing the susceptibility to secondary infections. Foot and mouth disease (FMD) is a highly contagious acute vesicular viral disease effectively controlled by vaccination in endemic regions. Despite the evidence of immunoregulatory effects, the impact of fasciolosis on the immune response induced by FMD vaccination in cattle has never been assessed. Our objective was to evaluate whether the infection by F. hepatica in cattle influences the long-term immunity elicited by the currently used commercial FMD-inactivated vaccines. Aberdeen Angus steers negative for F. hepatica were vaccinated twice against FMD virus (FMDV) during the first 6 months of age using a commercial oil vaccine formulated with A24/Cruzeiro and O1/Campos strains. When maternal antibodies against F. hepatica were weaned (18--20 months of age) animals were divided into groups of 12 and infected or mock-infected with 500 metacercariae/animal. Individual serum samples were collected at 0-, 28-, 59-, 87- and 157-days post-infection (dpi). Indirect ELISAs were used to detect A24/Cruzeiro specific bovine IgG and IgG subtypes. The total IgG antibody levels and avidity against FMDV did not show significant differences between all the groups. The commercial vaccine induced higher IgG2 than IgG1 titers in vaccinated animals. Anti-FMDV IgG1 levels significantly decreased in the infected group at 28 dpi. In addition, the avidity of IgG1 FMDV-specific antibodies at day 28 in the infected group was reduced compared to the control. These results show that F. hepatica infection modified anamnestic responses against FMDV, reducing serum IgG1 titers and avidity. To our knowledge, this is the first report of immune-regulation of F. hepatica altering the immune response of FMD vaccines, one of the most globally used animal vaccines.

Advances in diagnostic approaches to Fasciola infection in animals and humans: An overviews.

Fasciolosis, caused by Fasciola hepatica and F. gigantica, is an impediment to the livestock industry's expansion and has a massively negative socio-economic impact due to its widespread prevalence in livestock. It is a waterborne zoonosis affecting human populations in the countries where rural economies are associated with livestock rearing. Conventional diagnosis of Fasciola infection is done by detecting parasite eggs in the faeces of infected animals or by immunological methods. Accurate and quick immunodiagnosis of Fasciola infection in animals and humans is based on the detection of antibodies and specific antigens expressed in the prepatent stage of the parasite. Both molecular and serodiagnostic tests developed thus far have enhanced the reliability of Fasciola diagnosis in both man and animals but are not widely available in resource-poor nations. A pen-side diagnostic test based on a lateral flow assay or a DNA test like loop-mediated isothermal amplification (LAMP) would be simple, fast, and cost-effective, enabling clinicians to treat animals in a targeted manner and avoid the development of drug resistance to the limited flukicides. This review focuses on the recent advances made in the diagnosis of this parasite infection in animals and humans.

Geographic expansion of Fasciola hepatica (Linnaeus, 1758) due to changes in land use and cover in Brazil.

Fasciolosis is caused by parasites of the genus Fasciola, affecting animals and humans worldwide. In South America, the disease is a result of infection with Fasciola hepatica and although animal infections are more frequently reported, the full extent of the impact on human health due to underdiagnosis remains uncertain. This study analyzed changes in land use and the distribution of F. hepatica in bovine livers in Brazil over 18 years. Data on land use and land cover were collected from the Mapbiomas Project. Data on 414,481,963 slaughtered cattle and condemned livers due to F. hepatica infection were obtained from 4,433 municipalities. Joinpoint analysis was used to study the time series, and the Susceptible-Infected-Recovered (SIR) model was utilized to explore the behavior of F. hepatica infection. In the North, pasture areas significantly increased (P = 0.000001), while forested areas decreased (P = 0.000001). The midwestern and northern regions concentrated the highest number (>290 million) of cattle slaughtered in Brazil. More than 2 million bovine livers were infected by F. hepatica. The infected cattle originated from 194 municipalities in 2002, increasing to 747 in 2020. We consider that the changes in land use and intense cattle transportation may have caused the expansion of F. hepatica. The SIR model analyzed the spread of the disease looking at all six biomes: Caatinga, Amazon Forest, Cerrado, Pantanal, Atlantic Forest, and Pampa. Moreover, this infection not only threatens the health of animals but is also a major concern to biodiversity and vulnerable human communities in South America. Emblematic biomes such as the Amazon basin already face challenges with logging, desertification, and loss of biodiversity. Therefore, strategies for mitigating infection should include controlling illegal pasture areas, establishing health inspections of animal transport, quarantine of newly arrived animals, and livestock zoning, as well as clear One Health policies.

Prevalence and associated risk factors assessment of bovine fasciolosis in the Imbo Region, Burundi.

Fasciolosis is a zoonosis that limits the productivity of ruminants worldwide, but there is a lack of information on its occurrence in Burundi. Therefore, this study aimed to fill the information gap by determining the prevalence and risk factors associated with bovine fasciolosis in the Imbo Region of Burundi. Two prevalence studies were conducted in parallel in the five communes of the five provinces in the Imbo region. In the first study, a total of 426 fecal samples were collected from randomly selected cattle farms and microscopically examined to determine Fasciola egg burden. Survey data on cattle husbandry were collected from owners of these cattle and analyzed to determine the risk factors for bovine fasciolosis. In the second study, 467 cattle were randomly selected in abattoirs and their livers were examined postmortem to determine liver fluke burdens. Data were entered separately into Microsoft Excel and analyzed using R software. The overall prevalence of bovine fasciolosis was 47.7% (42.9-52.4, 95% CI) for microscopic examination and 33.2% (28.9-37.5, 95% CI) for postmortem examinations. The majority of positive cattle (60.6%) had light intensity infections as determined by eggs per gram of feces (epg). Postmortem examinations corroborated these results and indicated that 80% of cattle had light intensity infections. Chi-square analysis showed a statistical association with the presence of bovine fasciolosis and the age, sex, and origin of cattle and the practices of cattle owners (P < 0.05).

The Fasciola hepatica egg development and hatching test (EDHT) using commercial drugs: a simple protocol for tackling anthelminthic resistance and promoting cattle productivity.

Fasciolosis is an important zoonotic disease caused by the trematode Fasciola hepatica, and it causes great losses in bovine production. The anthelmintic resistance is a major problem in the control of fasciolosis. In this study, the F. hepatica egg development and hatching test (EDHT) was used for the evaluation of the ovicidal activity of commercial drugs, commonly used for treating infected cattle, which reflects F. hepatica anthelminthic resistance in infected bovines, according to recent literature. Bile samples from F. hepatica naturally parasitized cattle were obtained from slaughterhouses in the cities of Lages and Otacílio Costa, Santa Catarina State, Brazil. The bile was washed, the eggs were recovered, quantified, and distributed in universal collectors, with a minimum of 1,000 eggs per vial. Four commercial drugs were used in this study, containing albendazole sulfoxide (ABDZ), closantel (CSTL), nitroxynil (NTXL), and triclabendazole with fenbendazole (TBZF). The drugs were diluted according to the manufacturer instructions. All drugs, and the respective control, were tested in triplicates, with the quantity of recovered eggs determining the number of drugs to be tested. The vials were incubated for 28 days at 27 °C, and the eggs were classified according to their degree of development under a stereomicroscope. In total, 121 egg samples were analyzed. Two samples were identified as resistant to TBZF. Undetermined resistance/susceptibility has been found in two isolates treated with ABDZ, one treated with NTXL and six treated with TBZF. CSTL did not present ovicidal activity and cannot be used in EDHT. This is the first time that commercial drugs were used in F. hepatica EDHT.

Seroprevalence and risk factors of bovine fasciolosis in the municipalities of Colombia.

Background and Aim: Bovine fasciolosis is a reemerging neglected disease with a worldwide distribution caused by the trematode Fasciola spp., which parasitize various hosts. Bovine fasciolosis is responsible for large economic losses in the bovine livestock sector. This study aimed to estimate the seroprevalence and risk factors of bovine fasciolosis in the municipalities of Colombia. Materials and Methods: This was a descriptive cross-sectional study with simple random sampling conducted on 1140 cattle from the municipalities of Chiquinquirá, San Miguel de Sema, and Ubaté for a duration of 3 months. Serum samples were processed using the commercial Fasciola hepatica Antibody Test Kit IDEXX® Fasciolosis Verification (IDEXX, United States), which identified immunoglobulin G antibodies for gf2 antigen purified from Fasciola extracts. The f2 antigen is extremely immunogenic and highly specific for F. hepatica. An epidemiological survey was performed to record variables related to the sampled animals and herd management practices. Data were processed using the statistical program Epi Info® (Centers for Disease Control and Prevention; Atlanta, Georgia). The prevalence ratio was estimated to evaluate the association between fasciolosis and the hypothesized causal factors and the significance of this association using Pearson's Chi-square test. Finally, a logistic regression model was developed. Results: The overall seroprevalence was 72.3%. The seroprevalence was 83.9% (323/385) in Chiquinquirá, 68.17% (257/377) in Ubaté, and 64.55% (244/378) in San Miguel de Sema. The seroprevalence was higher in male animals in Chiquinquirá and in female animals in San Miguel de Sema and Ubaté. Similarly, sex showed a statistically significant association with disease prevalence in Ubaté. The highest prevalence was found in cattle aged >2 years. The Holstein breed showed maximum seroprevalence in Chiquinquirá (p ≤ 0.05) and San Miguel de Sema, whereas crossbreed showed higher seroprevalence in Ubaté. Similarly, in Chiquinquirá, the association between the seroprevalence of fasciolosis and the presence of other species was statistically significant (95% confidence interval [CI]: 0.9601-3.4944; p = 0.0448). In Ubaté, the disease presentation was also associated with pasture rental (95% CI: 0.4047-1.0023; p = 0.003) and attendance to livestock expositions (95% CI: 0.2313-1.0636; p = 0.044). However, in San Miguel de Sema, water from the stream showed a statistically significant association with disease presentation (95% CI: 0.5209-1.0985; p = 0.00649785). Female sex and diarrhea occurrence were considered risk factors for fasciolosis. Conclusion: A high seroprevalence of antibodies to Fasciola spp. was detected in cattle in the study municipalities, indicating a high parasite distribution in these areas. Female sex and diarrhea were established as risk factors associated with fasciolosis in Ubaté and San Miguel de Sema, respectively. Further, research is necessary to establish prevention and control programs against parasitosis.

A Rare Human Helminth Infection in Russia.

Currently, more than 500,000 cases of various helminthes in humans are reported annually in the Russian Federation. This figure may not reflect the true incidence of helminthes, as only nine separate nosological forms are compulsory notifiable. The rest of the species of detected helminthes are included in a separate category of "other helminthes" or "rare helminthes". The bulk of the latter is represented by the helminthes with a rate of incidence that does not exceed one case per 100,000 people. This review is based on data derived from publications in the Russian language, both from the Russian Federation and international, as well as data available from various health treatment facilities in Russia. These data largely cover the period of the 1990s-2010s. A total of 15 species of "rare helminthes" are described in this review: anisakiosis, capillariosis, clonorchosis, dioctophymosis, dipylidiosis, echinochasmosis, fasciolosis, gastrodiscoidosis (amphistomiosis), metagonimosis, metorchiosis, nanophyetosis, pseudamphistomosis, sparganosis (spirometrosis), strongyloidosis and trichostrongylosis. Details of their geographical distribution, clinical and epidemiological peculiarities, and the difficulties they pose in diagnosis are provided. The public health importance of "rare helminthes" in Russia at present and in the forthcoming years is stressed.

Coprological and postmortem assessment and economic significance of bovine fasciolosis in cattle slaughtered at Tarcha Municipal Abattoir, Southern Ethiopia.

Bovine fasciolosis caused by Fasciola hepatica and Fasciola gigantica, is a neglected tropical snail-borne trematode disease of cattle that poses an adverse effect on animals' health culminating in economic damage. Cross-sectional investigation on coprological and postmortem assessment and economic significance of bovine fasciolosis in cattle slaughtered at Tarcha Municipal Abattoir, Southern Ethiopia through simple random sampling techniques was conducted from December 2020 to August 2021. Out of 384 cattle examined, 20.3% and 29.94% were found harboring Fasciola through coprological examination and postmortem examination respectively. The leading cause of fasciolosis in the study area was Fasciola gigantica (53.9%) as compared to F. hepatica (46.1%). The investigation revealed that the sex, origin, and body condition of the cattle has a statistically significant effect on the prevalence of bovine fasciolosis. The odds of male animals infested with fasciolosis were 2.25 times higher than female cattle and those of poor body-conditioned cattle were three times higher than cattle with good body condition. In postmortem examination, an average of 6 flukes were found in a single infested liver and the mean fluke count was (10 ± 1.97) in severely infested livers while in lightly infested livers (3± 1.79). Considering postmortem examination as the gold standard for diagnosis of fasciolosis, the sensitivity of the direct sedimentation technique was found to be 68% and the specificity 100% with substantial agreement (k = 0.74) between the two methods. The estimated yearly direct financial losses incurred owing to fasciolosis was around 2,227,536 2, Birr (47,945.24 USD). Thus, bovine fasciolosis is one of the economically important animal diseases in the study area, which necessitates integrated control measures to tackle its effect on animal health and subsequent economic impact.

Plasma lipopolysaccharide elevations in cattle associated with early-stage infection by Fasciola hepatica.

Fasciolosis is an endemic zoonotic parasitic disease with significant impacts on human health and both animal health and production. Early post-infection impacts on the host remain unclear. The objective of this study was to determine the changes, if any, to levels of endotoxin in cattle plasma in response to early-stage infection with Fasciola hepatica. Thirty-six (36) commercial bred cattle were experimentally infected with approximately 400 viable metacercariae. Plasma lipopolysaccharide (endotoxin) levels were examined on 24 occasions from 0 h before infection to 336 h after infection using the Limulus Amoebocyte Lysate chromogenic end point assay and compared with that of six (6) uninfected control animals. Peak lipopolysaccharide levels in infected animals were reached at 52 h after infection and returned to pre-infection levels at time 144 h after infection. Infected animals had significantly elevated lipopolysaccharide levels between 24 and 120 h after infection when compared to uninfected animals. The mean change in endotoxin units (EU)/mL over time after infection was statistically significant in infected animals. Elevations of lipopolysaccharide occurred in all infected animals suggesting a possible repeatable and titratable endotoxemia conducive to therapeutic agent model development.

Fascioliasis associated with chronic cholecystitis in a woman from Sistan and Baluchestan province, a non-endemic region in Southeastern Iran.

BACKGROUND: Fascioliasis, caused by Fasciola hepatica, is a neglected zoonotic food-borne trematodiasis. The Caspian littoral in northern Iran is endemic for the disease, and human fascioliasis is well-known in that region. In the present study, we report the diagnosis, identification, and clinical management of a human case of fascioliasis associated with common bile duct (CBD) obstruction from a non-endemic remote area in southeastern Iran. CASE PRESENTATION: A 42-year-old female was admitted to Afzalipour Medical Center hepatobiliary surgery ward in Kerman with abdominal pain for the past three months. Dilated biliary tract and an ill-defined mass in CBD were reported in abdominal ultrasonography and magnetic resonance cholangiopancreatography, respectively. During distal CBD operation, nine leaf-like motile flatworms were isolated. A morphological study confirmed all the isolates as Fasciola, and further molecular investigations, identified the flukes as F. hepatica using both pepck multiplex PCR and cox1 sequencing. CONCLUSION: Molecular and morphological findings of the study indicated the presence of human fascioliasis in the southeastern province of Sistan and Baluchestan in Iran. Fascioliasis is among the etiologies of chronic cholecystitis, and physicians should consider chronic cholecystitis associated with fascioliasis in the differential diagnosis. In the present report, endoscopic ultrasound was usefully applied for the accurate diagnosis of biliary fasciolosis.

Molecular Characterization of the Interplay between Fasciola hepatica Juveniles and Laminin as a Mechanism to Adhere to and Break through the Host Intestinal Wall.

Fasciola hepatica is the main causative agent of fasciolosis, a zoonotic parasitic disease of growing public health concern. F. hepatica metacercariae are ingested by the host and excyst in the intestine, thereby releasing the newly excysted juveniles (FhNEJ), which traverse the gut wall and migrate towards the biliary ducts. Since blocking F. hepatica development is challenging after crossing of the intestinal wall, targeting this first step of migration might result in increased therapeutic success. The intestinal extracellular matrix (ECM) is constituted by a network of structural proteins, including laminin (LM) and fibronectin (FN), that provide mechanical support while acting as physical barrier against intestinal pathogens. Here, we employed ELISA and immunofluorescent assays to test for the presence of LM- and FN-binding proteins on a tegument-enriched antigenic fraction of FhNEJ, and further determined their identity by two-dimensional electrophoresis coupled to mass spectrometry. Additionally, we performed enzymatic assays that revealed for the first time the capability of the juvenile-specific cathepsin L3 to degrade LM, and that LM degradation by FhNEJ proteins is further potentiated in the presence of host plasminogen. Finally, a proteomic analysis showed that the interaction with LM triggers protein changes in FhNEJ that may be relevant for parasite growth and adaptation inside the mammalian host. Altogether, our study provides valuable insights into the molecular interplay between FhNEJ and the intestinal ECM, which may lead to the identification of targetable candidates for the development of more effective control strategies against fasciolosis.

The First Case of Human Hepatic Fasciolosis Presented as Hepatic Pseudotumor Histopathologically Diagnosed in Romania-A Case Report.

Human hepatic fasciolosis has been reported in 81 countries, some of which are endemic areas. In Europe, case reports from humans were published in Portugal, Spain, France, and Italy. Regarding Romania, we do not have any data on the prevalence of this parasitosis, with the exception of two cases of twins that were born in Romania and diagnosed in the last 37 years in Italy after joining their mother that lived there. We report the case of a patient diagnosed in Romania with chronic fasciolosis, presented as a hepatic pseudotumor that was diagnosed during the histopathological examination of the hepatic lesion. The patient received oral treatment with triclabendazole, two doses of 10 milligrams (mg) per kilogram (kg) of body weight, given 12 h apart, with no side effects during or after the treatment. The evolution of the patient was favorable. In conclusion, even in areas free of human fasciolosis, the presence of an anemic syndrome especially in children, abdominal pain in the upper quadrants, associated or not with other digestive manifestations, even more so associated with eosinophilia in the acute phase, should be carefully evaluated for ruling out a parasitosis such as fasciolosis even in countries where this diagnosis seems unlikely.

Prevalence and Risk Factors of Ovine and Caprine Fasciolosis in the Last 20 Years in China: A Systematic Review and Meta-Analysis.

Fasciolosis is a significant zoonotic and common parasitic disease for animals and humans, creating public health concerns worldwide. This study retrieved articles related to the occurrence of Fasciola hepatica and Fasciola gigantica in sheep and goats in China by searching five databases: PubMed, ScienceDirect, the Chinese National Knowledge Infrastructure (CNKI), Wanfang Data, and the VIP Chinese Journal Database. A total of 60 valid articles were captured. The pooled prevalence of ovine and caprine fasciolosis was 26.00%. It was also found to be higher in the subgroups of Northwest China and Shaanxi Province, as well as in areas with a high altitude, rainfall of ≥800 mm, and temperature ranging between 10 °C and 20 °C. Analysis of the type of season and sampling years showed significant (p < 0.05) difference. In other subgroups, sheep (34.74%), hosts aged over 2 years (32.26%), females (48.33%) and free-range animals (26.83%) showed a higher disease prevalence. These results indicated that ovine and caprine fasciolosis was widely distributed, especially in Northwest China. The sampling years and the type of season are risk factors for the prevalence of ovine and caprine fasciolosis. Therefore, strategies for ovine and caprine fasciolosis control should be developed based on these epidemic risk factors, which will reduce the prevalence of fasciolosis in China.

Prevalence of Human and Animal Fasciolosis in Butajira and Gilgel Gibe Health Demographic Surveillance System Sites in Ethiopia.

Fasciolosis is regarded as a major challenge to livestock productivity worldwide, but the burden of disease in humans has only started to receive some attention in the past three decades. The aim of this study was to determine the prevalence of human and animal fasciolosis and its determinant factors in the Gilgel Gibe and Butajira Health and Demographic Surveillance System (HDSS) sites in Ethiopia. A study was undertaken among 389 households across the two sites. Face-to-face interviews were conducted to investigate the knowledge, attitudes and practices of households with regard to fasciolosis. Stools from 377 children aged 7-15 years, and 775 animals (cattle, goats and sheep) were analyzed using a proprietary Fasciola hepatica (F. hepatica) coproantigen ELISA kit. The prevalence of fasciolosis in children was 0.5% and 1% in Butajira and Gilgel Gibe HDSS sites, respectively. The overall prevalence of animal fasciolosis was 29%, 29.2%, and 6% among cattle, sheep, and goats, respectively. More than half of the respondents from Gilgel Gibe (59%, n = 115) did not know that humans can be infected with F. hepatica. The majority of respondents in Gilgel Gibe (n = 124, 64%) and Butajira (n = 95, 50%) did not know the transmission route for fasciolosis. Grazing animals were 7 times more likely to be infected with fasciolosis than animals in cut-and-carry production systems (adjusted odds ratio [AOR] = 7.2; 95% confidence interval [CI]: 3.91-13.17). The findings indicated a lack of knowledge amongst local populations about fasciolosis. Thus, there is a need for public health awareness campaigns about fasciolosis in the study areas.

Development of an enzyme linked immunosorbent assay using recombinant cathepsin B5 antigen for sero-surveillance of bovine tropical fasciolosis.

Bovine tropical fasciolosis, caused by Fasciola gigantica, is a major parasitic disease in tropical countries responsible for significant production losses in animal husbandry practices. The disease is transmitted by the Radix sp. snails. In the early developmental stage of the parasite, the juveniles and immature flukes cause considerable damage to the liver parenchyma of the bovine host while migrating through the liver. The cathepsin (cat) B5 is a cysteine protease that is present in the excretory-secretory product of the fluke both in immature and adult stages. The early detection of fasciolosis is very critical in effective disease management. In this study, the cathepsin B5 gene from newly excysted juveniles were cloned, sequenced and analyzed. The phylogenetic analysis revealed existence of two distinct clades. The clade I includes cat B 1 to B3 whereas clade II consist of cat B4 to B7. Further, the present study was aimed to develop an enzyme linked immuno sorbent assay (ELISA) using recombinant cat B5 antigen. The developed enzyme immuno assay showed 95.3 % sensitivity and 92.4 % specificity with a cut-off of 60 % percent positive. It revealed weighted Kappa value as 0.768 (95 % CI 0.648-0.889) when compared with ELISA using native cathepsin protein. Hence, the developed assay can be exploited as a potent tool in the diagnosis and sero-surveillance of bovine tropical fasciolosis.

Comparison of the therapeutic efficacy of five anthelmintics against natural Fasciola hepatica infections in dairy cattle from the Mantaro Valley, Peru.

The intensive use of anthelmintic drugs to control Fasciola hepatica infections in dairy cattle has resulted in the emergence of anthelmintic resistance. Cases of resistance to triclabendazole (TCBZ) have been reported worldwide. The main goal of this research was to evaluate the main five fasciolicides to control fasciolosis in dairy cattle in the Mantaro Valley, Peru. Two fecal egg count reduction tests were performed. In a first study, 24 naturally F. hepatica infected cattle were randomly grouped into three experimental groups (n = 8). Groups were treated with either TCBZ, nitroxynil (NTX) or closantel (CLOS). In a second experiment, 55 naturally infected cows were grouped into three experimental groups and treated with either TCBZ (n = 18), rafoxanide (RFX) + albendazole (ABZ) (n = 19) or clorsulon (CLN) + ivermectin (IVM) (n = 18). Therapeutic efficacy was determined following the WAAVP guidelines by measuring reduction in fluke egg output at days 15 and 30 post-treatment. Bootstrapping method was used to obtain the 95% confidence intervals. The efficacy of TCBZ was inadequate in both studies (≤80.8%). Closantel showed high efficacy (≥ 90%) at both days, while NTX showed 92.9% (83-100) and 82.1% (53.6-100), efficacy, at days 15 and 30, respectively. Efficacy for RFX were 92.1% (79.6-98.9) and 97.4% (94.1-99.4); and for CLN, 98.8% (97.6-100) and 80.1% (44.7-99.4), at days 15 and 30, respectively. The outcome of this study indicates reduced therapeutic efficacy of TCBZ against F. hepatica in an important dairy area of the Peruvian central highlands but also demonstrates the validity of four alternatives.

First Molecular Identification of Fasciola gigantica in Slaughtered Cattle in Cape Verde: Prevalence, Gross Pathological Lesions, Genetic Identification and Coprological Analysis.

A study on fasciolosis prevalence, gross pathological lesions, fluke genetic identification and coprological analysis was carried out in slaughtered cattle from one abattoir in Cape Verde. Of the 131 cattle inspected over two months, 12 (9.0%) presented fasciolosis-compatible lesions (FCL) that resulted in liver condemnation. The genetic characterization of the flukes collected, through restriction fragment length polymorphism analysis of PCR-amplified fragments (PCR-RFLP), confirmed the presence of Fasciola gigantica; therefore, being the first identification of this species in cattle from Cape Verde. Animals that released Fasciola spp. eggs and, thus, responsible for environment contamination (positive shedders), were identified through coprological analysis (natural sedimentation technique). Of the 12 animals with FCL, samples from 11 were submitted to coprological analysis and 7 (63.6%) were found to be positive shedders. Furthermore, of the 82 animals with non-FCL, randomly selected for coprological analysis, 4 (4.9%) were also found to be positive shedders for Fasciola spp. The results of this study, regarding species identification and coprological analysis, are epidemiologically important to update the information regarding fasciolosis in Cape Verde. The new data could help implement effective strategies for disease control and mitigation, consequently reducing economic loss and the level of animal and human infection from the One Health perspective.

Fasciola gigantica vaccine construct: an in silico approach towards identification and design of a multi-epitope subunit vaccine using calcium binding EF-hand proteins.

Continuous attempts have been made to pinpoint candidate vaccine molecules and evaluate their effectiveness in order to commercialise such vaccines for the treatment of tropical fascioliasis in livestock. The pathophysiology of fascioliasis can be related to liver damage brought on by immature flukes that migrate and feed, as well as immunological reactions to chemicals produced by the parasites and alarm signals brought on by tissue damage. Future research should, in our opinion, concentrate on the biology of invasive parasites and the resulting immune responses, particularly in the early stages of infection. The goal of the current study was to use the calcium-binding proteins from F. gigantica to create a multi-epitope subunit vaccine. The adjuvant, B-cell epitopes, CTL epitopes, and HTL epitopes that make up the vaccine construct are all connected by certain linkers. The antigenicity, allergenicity, and physiochemical properties of the vaccine construct were examined. The vaccine construct was docked with toll-like receptor 2, and simulations of the molecular dynamics of the complex's stability, interaction, and dynamics were run. After performing in silico cloning and immunosimulation, it was discovered that the construct was suitable for further investigation. New vaccination technologies and adjuvant development are advancing our food safety procedures since vaccines are seen as safe and are accepted by the user community. This research is also applicable to the F. hepatica system.