Search

Background: Fasciola gigantica is a helminth parasite of veterinary importance and a constraint on the growth and productivity of domestic livestock in the tropical countries of Asia and Africa. Present study aimed at determining the expression of four cysteine protease genes including cathepsin (cat) L-1 D and cat-Bs in the embryonated egg stage of F. gigantica by polymerase chain reaction (PCR). In addition to the analysis of expression of the above genes in the embryonated eggs, inhibition of in vitro hatching of the miracidia from the eggs was carried out using three protease inhibitors viz. serine, cysteine and leucine aminopeptidase inhibitors to understand the possible role of these proteases in the hatching of the miracidia from the eggs. Methods: Cat- L-1D, cat B-1, cat B-2 and cat B-3 genes were amplified by reverse transcriptase and quantitative-PCR, respectively for determining their expression in the embryonated eggs. Inhibition of in vitro hatching of the miracidia from the eggs was carried out by exposing the embryonated eggs to different concentrations of iodoacetamide, phenyl methyl sulphonyl fluoride and bestatin as cysteine, serine and leucine aminopeptidase inhibitors. Result: Results showed that cat L1-D, cat B-1 and cat B-3 genes were expressed at the embryonated egg stage but cat B-2 gene was not expressed at this stage of the parasite. Iodoacetamide and phenyl methyl sulphonyl fluoride (cysteine and serine protease inhibitors) inhibited the miracidial hatching at different concentrations, indicating probable role of cysteine and serine proteases in its hatching process. However, leucine aminopeptoidase does not seem to influence the miracidial hatching from the eggs as bestatin did not inhibit its hatching. [ABSTRACT FROM AUTHOR]

Background: Bovine anaplasmosis caused by Anaplasma marginale is highly endemic in tropical and sub-tropical regions of the world where it accounts for huge economic losses to the livestock sector. Approximately 30% of water buffalo (Bubalus bubalis) herds in India are located in Uttar Pradesh (U.P). However in India, most of the research studies on Anaplasma are focused on cattle whereas studies on this pathogen in water buffaloes have been neglected. Therefore, present investigation was undertaken for determining the prevalence of A. marginale infection in water buffaloes in U.P, North India. Methods: In the current investigation during 2017-2019, the prevalence of A. marginale infection in 228 water buffaloes in three districts of North-West U.P was studied by microscopy and by nested-PCR amplification of major surface protein 5 (msp5) gene. Result: Microscopic examination of blood smears of 228 water buffaloes revealed 7.89% animals positive for A. marginale. Conventional PCR of full length major surface protein 5 (msp5) gene detected 29.39% and nested-PCR amplification of this gene detected 64.0% of these animals as positive for A. marginale infection. A high incidence of the infection detected in this study area suggests that water buffaloes may serve as reservoir hosts of A. marginale and play a vital role in the spread of infection to other susceptible animals. However further studies are required on larger buffalo populations for determining the epidemiological status of anaplasmosis in these animals in India. [ABSTRACT FROM AUTHOR]

Fasciola gigantica cathepsin (cysteine) proteases are potential diagnostic antigens for animal and human fasciolosis. These include cathepsin-L proteases that have been exploited in the diagnosis of animal fasciolosis. However, no scientific data on the diagnostic potential of F. gigantica cathepsin B proteases is available. Therefore, three recombinant antigens of F. gigantica viz. cathepsin (cat) B-1, cat B-2 and cat B-3 were expressed in prokaryotic expression system. The recombinant antigens were purified under denaturing conditions by Nickel affinity chromatography and an optimal level of the recombinant proteins was obtained. These recombinant proteins will further be evaluated for their potential in the early prepatent diagnosis of F. gigantica infection in domestic ruminants. [ABSTRACT FROM AUTHOR]

Mitochondrial DNA sequence of the sub-unit 1 of cytochrome c oxidase (Cox1) and ribosomal DNA sequence of the first internal transcribed spacer (ITS-1) of Fasciola collected from mithun and cattle from Arunachal Pradesh, India were characterized at molecular level in order to identify the species of parasite. Based on sequence and phylogenetic analysis, the identity of the parasite was confirmed as Fasciola gigantica in mithun as well as from cattle. In order to know histopathological alteration in Fasciola infection, a histopathological study was performed on eight liver specimens, out of 110 animals studied during the period from 2010-2016. Histopathological examination of F.gigantica infected liver showed extensive fibrous connective tissue proliferation with necrosis of hepatocytes and infiltration of polymorphonuclear cell. There was evidence of migratory tracts of parasites with losing of normal lobular hepatic architecture. Bile duct proliferation was followed by congestion of portal vein with perivascular cuffing with surrounding degeneration of hepatocytes. The hepatocytes showed pyknosis with hyperplasia of bile duct. [ABSTRACT FROM AUTHOR]

Parasitism is a common problem for mithun (Bos frontalis) resulting in production losses in terms of meat and milk. Prevalence of ectoparasites infestation including Rhipicephalus microplus, Ambylomma testuidinarium, Ixodes ovatus and I. acutitarsus, has been reported based on taxonomic studies and PCR amplification of conserved gene. Different gastrointestinal helminth like Haemonchus contortus, Mecistocirrus digitatus, Trichostrongylus spp., Oesophagostomum spp., Trichuris spp s, Thelazia species, Strongyloides papillosus, different amphistomes and Fasciola gigantica were also reported from mithun from India. Among filarial worms, Setaria digitata was recorded from peritoneal cavity of mithun and its molecular confirmation was done based on PCR amplification of marker genes. Tapeworm infestation viz. Moniezia expansa, M. benedeni, hydatid cyst and Cysticercus tenuicollis, were also commonly occurred in mithun. Enteric protozoa like different species of Eimeria, Cryptospordium species, Balantidium coli, has been diagnosed based on morphological identification sporulated oocysts and developmental stages found on faecal examination. Another obligate intracellular parasites Toxoplasma gondii, was also diagnosed based on ELISA and modified agglutination test. An integrated approach incorporating deworming, good livestock management and optimum feeding management practice coupled with vaccination programme will make successful parasite control programme in near future. [ABSTRACT FROM AUTHOR]

Echinococcus granulosus recombinant proteins including two antigen B subunits EgAgB8/1 and EgAgB8/2 and Echinococcus protoscolex calcium binding protein (EPC1) were expressed in prokaryotic expression vectors. The diagnostic potential of these three recombinant proteins was evaluated in the detection of cystic echinococcosis in goats in IgG-ELISA. The EgAgB8/1 and EgAgB8/2 recombinant proteins reacted fairly with the hydatid infected goats with sensitivity of 66.7% and 80.0% and specificity of 71.3% and 73.3%, respectively while EPC1 recombinant protein showed lower sensitivity (60%) but comparable specificity (72.3%). Cross-reactivity of these three antigens with goat gastro-intestinal strongyle nematodes and Taenia hydatigena under field conditions was studied. Results showed that EgAgB8/1, EgAgB8/2 and EPC1 antigens cross-reacted with most of the parasites in the goat host. [ABSTRACT FROM AUTHOR]

The article discusses the study on the prevalence of Cryptosporidium spp. infections, that involves the testing of faecal samples of diarrhoeic bovine calves in dairy farms in Uttar Pradesh, Uttarakhand and Bihar, India. The 25 Cryptosporidium positive samples, based on the Polymerase Chain Reaction Restriction Fragment Length Polymorphism (PCR-RFLP) patterns of 18 ribosomal ribonucleic acid (rRNA), were identified to be positive for C. parvum.

The article discusses a study that examined the primer-directed amplification of the open reading frame (ORF) of the GRA5 gene of two Indian isolates of the Toxoplasma gondii, namely Izatnagar (IZN) and Chennai (CHEN). It details the molecular cloning, comparative sequence analysis and prokaryotic expression of mature GRA5 protein. It highlights the purification and the results of Western blot analysis of expressed histidine-tagged recombinant mature GRA5 protein.

Fasciola gigantica fatty acid binding protein (FABP) was evaluated for evoking an immune response in mice, by delivering the gene coding for this protein with mannosylated-polyethylenimine (PEI) to peritoneal cells. Mice were immunized with 50 µg recombinant plasmid DNA (Group I) or DNA-PEI-mannose (a 22 kDa linear cationic polymer with mannose ligand) (Group II) via the intraperitoneal route. Antibody studies showed no significant humoral immune response evoked to this DNA immunization with either PEI-mannose-delivered or naked DNA. However, on protein boosting of these DNA-primed mice there was a significant enhancement of antibody titre. Flow cytometric bead array was used to measure quantitites of interleukin (IL)-2, IL-4, IL-5, interferon-γ (IFN-γ) and tumour necrosis factor (TNF) cytokines. Overexpression of T-helper 1 (Th1) cytokines such as IFN-γ and TNF, with a lower but significant expression of the T-helper 2 (Th2) cytokine IL-5 was detected. Gene delivery using polyethylenimine-mannose ligand showed significant expression of IFN-γ and TNF (P < 0.05), but no significant difference in IL-2, IL-4 and IL-5 (P > 0.05) cytokine expression was observed between naked-DNA- and mannosylated PEI-DNA-delivered mice. Naked- or PEI-delivered-DNA immunization produced insignificant levels of IL-2 and IL-4 (P > 0.05) cytokines in both groups of mice. [ABSTRACT FROM AUTHOR]

Glutathione S-transferase from an Indian isolate of Fasciola gigantica of buffalo origin was isolated and characterized. Total RNA was transcribed to cDNA by reverse transcription and an amplicon of 657 bp glutathione S-transferase gene was obtained by polymerase chain reaction (PCR). The present isolate showed 99.1% sequence homology with the published sequence of the F. gigantica GST gene of cattle origin, with six nucleotide changes causing an overall change of four amino acids. Glutathione S-transferase protein was expressed in Escherichia coli using a prokaryotic expression vector pPROEXHTb. The recombinant protein was purified under non-denaturing and denaturing conditions by nickel nitrilotriacetic acid (Ni-NTA) affinity chromatography. Recombinant GST protein detected F. gigantica infection in naturally infected buffaloes by dot-ELISA. [ABSTRACT FROM AUTHOR]

The article discusses a research study which examined the effect of deoiled mahua seed cake (DMSC) extract on juvenile flukes of Fasciola gigantica. Data shows the mortality of newly excysted juvenile flukes of F. gigantica at hourly intervals exposed to aqueous extracts of deoiled mahua seed cake. Phytochemical analysis revealed the total tannins and saponins content in DMSC.

The article deals with the use of protoscolices (PSCs) antigens for fast detection of hydatidosis in buffaloes in India. The procedure for preparing antigens of buffalo origin is discussed. The potential of such antigens for detection of antibodies in buffaloes naturally infected with the disease was evaluated by performing the counter immunoelectrophoresis (CIEP). Also noted is the possible importance of PSCs in the immunodiagnosis and protection.

The article discusses a study which deals with antibody responses following development of secondary hydatidosis in mice using surface antigens of protoscolices of buffalo origin extracted via common biological detergents, sodium deoxycholate and TritonX-100. Similar trend of antibody response was observed in both the surface antigen fractions of protoscolices. Results suggest the potential use of surface antigens of protoscolices for immunodiagnosis.

Aim: The objective of this study was characterize the somatic soluble antigens of adult Toxocara canis (Tc-SA) by western blotting. Materials and Methods: T. canis worms were collected from the naturally infected pups after deworming. The somatic antigen was prepared as per standard procedure with slight modification. These antigens were separated using Sodium dodecyl sulphate-electrophoresis (SDS-PAGE). The specific reactivity of the Tc-SA proteins was checked against the serum of naturally infected dogs as well with the hyperimmune serum raised in the rabbit by western blotting. Results: On SDS-PAGE recovered proteins ranged in size from 44 to 300 kDa. The immuno-reactivity of the naturally infected dog sera with the Tc-SAantigens showed 12 prominent immunoreactive bands of distinct sizes at 28.61, 32.60, 38.10, 43.04, 49.99, 67.57, 73.22, 105.77, 144.74, 161.11, 177.84 and 196.31 kDa. The immuno-reactivity of the hyper immune serum raised in rabbits against Tc-SA antigens was observed with 10 prominent bands of distinct sizes at 17.11, 24.15, 34.83, 43.46, 52.47, 55.89, 67.57, 70, 74.60 and 105.6 kDa. Conclusions: Common antigens band were observed at 67 and 105 kDa. These antigens merit further evaluation as candidate for use in diagnosis of toxocarosis in humans and adult dogs. [ABSTRACT FROM AUTHOR]

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.

Fasciola gigantica faces a series of threats from various free radicals produced by the host immune system during its invasion through the abdominal cavity and establishment in the bile duct of ruminants, limiting the fluke viability. The role of the superoxide radical produced by Muzaffarnagari sheep immune effector cells against F. gigantica newly excysted juveniles (NEJs) is highlighted in this study, as is the critical role of superoxide dismutase enzyme (SOD) in dismutation of superoxide radicals derived from host immune effector cells in vitro. Three concentrations of the ovine immune effector cells viz. 2.5, 5, and 10 × 10 6  cells were tested for their ability to induced cytotoxic killing of the parasite. All the three cell concentrations caused significant (p < 0.01) cytotoxic killing of NEJs in comparison to the control groups. Also, reduction of the immune effector cell concentration directly correlates with the NEJs killing. Attachment of immune effector cells to the parasite tegument in the presence of anti-F. gigantica antibodies was found to be critical in inducing NEJs killing via antibody-dependent cell-mediated cytotoxicity (ADCC). However, the addition of SOD greatly inhibits cytotoxic killing of NEJs, demonstrating the importance of SOD enzyme in fluke survival and parasite evasion of the host immunity. Thus, F. gigantica SOD warrants a promising candidate for immunoprophylactic studies in ruminants against the tropical liver fluke., (Copyright © 2022 Elsevier Inc. All rights reserved.)

Anaplasma marginale infection in cattle (n = 216) in the states of Uttar Pradesh and Uttarakhand, North India was screened by microscopy and nested-polymerase chain reaction (PCR). Two recombinant proteins viz. major surface protein (MSP) 5 and MSP2 of A. marginale were expressed in Escherichia coli and their potential in the detection of antibodies to Anaplasma species in the cattle was evaluated by immunoglobulin G-enzyme linked immunosorbent assay (IgG-ELISA). The MSP5 IgG ELISA results were compared with competitive (c) inhibition ELISA. Microscopy being the least sensitive diagnostic test detected 12.0% of animals positive for A. marginale infection while nested-PCR detected 87.9% of these animals as positive for A. marginale infection. The recombinant MSP5 antigen showed positive reactivity in 170/190 nested-PCR confirmed positive animals (sensitivity 89.5%) with specificity of 77.0%. In comparison, the recombinant MSP2 antigen showed lesser sensitivity and specificity of 79.0% and 69.2%, respectively. The cELISA was more sensitive and specific than IgG-ELISA. However, molecular detection by msp5 nested-PCR was highly sensitive and reliable for detection of carrier cattle for Anaplasma infection. The study indicated that a large cattle population (87.9%) was carrier for A. marginale infection in this region of the country., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Fasciolosis in ruminants is a relentless constraint in the livestock industry across the world. Immuno-prophylactic vaccines against fasciolosis may not come up in near future, rendering the control of this scourge with chemotherapy and snail population control. With the alarming threats of anti-fasciolid drug resistance reported from certain parts of the world; the control of fasciolosis should be directed towards the development of rapid and reliable diagnostic tools to execute the specific and discrete treatment. Understanding the epidemiology of Fasciola, its genomics and proteomics, host-parasite interplay, and advances in drug design research is vital for improving animal health that would ultimately succour to meet the ever-increasing demand for food. Due to possible differences in immune response depending on the species of the host and parasite, immuno-prophylactic studies in India should aim at achieving protective efficacy in buffalo against F. gigantica as workers from other countries concentrate primarily on vaccination of cattle and sheep against F. hepatica. This manuscript focused on the research that has been carried out in India for understanding the epidemiology, genetic diversity, immuno-diagnosis, and possible control measure in terms of immuno-prophylaxis and drug designing against tropical fasciolosis caused by Fasciola gigantica., (Copyright © 2021 Elsevier Inc. All rights reserved.)

Three recombinant proteins of Echinococcus granulosus including two antigen B sub-units EgAgB8/1 and EgAgB8/2 and Echinococcus protoscolex calcium binding protein 1 (EPC1) were expressed in prokaryotic expression vectors. The diagnostic potential of these three recombinant proteins was evaluated in the detection of cystic echinococcosis in buffaloes in IgG-ELISA. The EgAgB8/1 and EgAgB8/2 recombinant proteins reacted fairly with the hydatid infected buffaloes with sensitivity of 75.0% and 78.6%, respectively and specificity of 75.8% while EPC1 recombinant protein showed higher sensitivity (89.3%) but lower specificity (51.5%). Cross-reactivity of these three antigens was assayed with buffalo sera naturally infected with Explanatum explanatum, Paramphistomum epiclitum, Gastrothylax spp., Fasciola gigantica and Sarcocystis spp. EgAgB8/1 and EPC1 antigens cross-reacted with all these sera while EgAgB8/2 showed no cross-reaction with Sarcocystis spp. and reacted with some of the E. explanatum infected buffalo sera. This study explores the potential of three hydatid antigens viz. EgAgB8/1, EgAgB8/2 and EPC1 for their diagnostic potential in buffaloes positive for cystic echinococcosis., (Copyright © 2018 Elsevier B.V. All rights reserved.)