Fasciolosis, caused by
and
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
infection is done by detecting parasite eggs in the faeces of infected animals or by immunological methods. Accurate and quick immunodiagnosis of
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
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.
Highlights ► First comprehensive review regarding edible veterinary parasitic vaccines. ► Transgenic plant based edible vaccines possess exceptional merits. ► Trials on plant based vaccines against ...significant parasites bestow promising results. ► Immunization via antibodies expressed in plants is yet another innovative strategy.
Fasciolosis in India: An overview Lalrinkima, H.; Lalchhandama, C.; Jacob, Siju Susan ...
Experimental parasitology,
March 2021, 2021-Mar, 2021-03-00, 20210301, Letnik:
222
Journal Article
Recenzirano
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.
•Epidemiology fasciolosis in animal and human in India and genetic diversity is described.•Genetic diversity of Fasciola sp. in India is described.•Research on immuno-diagnosis, immuno-prophylaxis and potential drug identification in silico which have been carried out in India is highlighted.
Th1 and Th2 cytokine gene expression in buffalo calves during primary infection with Fasciola gigantica as well as in response to immunization with the parasite recombinant fatty acid binding protein ...(rFABP) and recombinant glutathione S-transferase (rGST) proteins was measured at 14th week of infection by real-time PCR with the double-stranded DNA-binding dye SYBR Green. Experimental animals were randomly distributed into FABP, GST, cocktail, challenge and healthy groups. Animals in groups FABP and GST were immunized with 400 μg rFABP and rGST, respectively, and cocktail group with a mixture of 400 μg each of rFABP and rGST in the neck and thigh muscles. All animals received three immunizations at 3-week interval. Calves were challenged per os with 400 viable metacercariae along with the unimmunized challenge control group 1 month after the last immunization. Expression of various cytokines in response to the immunization and parasite primary infection was measured by real-time PCR. Expression of IL-2 (4.5-fold) and IFN-γ (3.2-fold), followed by IL-6 (1.7-fold) and IL-4 (1.6-fold), with downregulation of TNF-α and IL-10 was observed in response to F. gigantica infection in these animals. However, there was a sharp increase in the expression of the IL-4 (211.93 and 111.81-fold) and IL-6 mRNA (219.22 and 48.29-fold) to GST and FABP immunizations, respectively. A downregulation of the IL-1α, a Th1 cytokine in response to FABP and GST immunization in these calves, was also observed. Overall, a mixed type of Th1 and Th2 cytokine environment was evoked to chronic F. gigantica infection and immunization with the above two recombinant proteins in buffaloes.
Babesia gibsoni is a tick borne intraerythrocytic protozoan parasite causing piroplasmosis in dogs and has been predominantly reported in Asian countries, including Japan, Korea, Taiwan, Malaysia, ...Bangladesh and India. The present communication is the first evidence on the genetic diversity of B. gibsoni of dogs in India. Blood samples were collected from 164 dogs in north and northeast states of India and 13 dogs (7.9%) were found positive for B. gibsoni infection by microscopic examination of blood smears. Molecular confirmation of these microscopic positive cases for B. gibsoni was carried out by 18S rRNA nested-PCR, followed by sequencing. Nested-PCR for the 18S rRNA gene was also carried out on microscopically B. gibsoni negative samples that detected a higher percentage of dogs (28.6%) infected with B. gibsoni. Genetic diversity in B. gibsoni in India was determined by studying B. gibsoni thrombospondin-related adhesive protein (BgTRAP) gene fragments (855bp) in 19 isolates from four north and northeast states of India. Phylogenetic analysis of the BgTRAP gene revealed that B. gibsoni parasite in India and Bangladesh formed a distinct cluster away from other Asian B. gibsoni isolates available from Japan, Taiwan and Korea. In addition, tandem repeat analysis of the BgTRAP gene clearly showed considerable genetic variation among Indian isolates that was shared by B. gibsoni isolates of Bangladesh. These results suggested that B. gibsoni parasites in a different genetic clade are endemic in dogs in India and Bangladesh. Further studies are required for better understanding of the genetic diversity of B. gibsoni prevalent in India and in its neighbouring countries.
•First evidence on genetic diversity in Babesia gibsoni of dogs in India•BgTRAP gene showed genetic diversity in the Indian isolates of B. gibsoni•Tandem repeats of consensus nucleotide sequences in the TRAP gene lead to genetic variations•B.gibsoni parasites in a different genetic clade are endemic in dogs in India and Bangladesh
Fasciola gigantica leucine aminopeptidase protein (56kDa) expressed in E. coli, characterized and evaluated for its vaccine potential along with recombinant peroxiredoxin (24kDa) in buffaloes. Both ...proteins failed to give significant level of protection in buffaloes. Display omitted
► Leucine aminopeptidase gene was identified in Fasciola gigantica. ► Recombinant leucine aminopeptidase did not evoke protective response in buffaloes. ► Peroxiredoxin recombinant protein in combination with leucine aminopeptidase also failed to evoke protection in buffaloes.
Gene coding for leucine aminopeptidase (LAP), a metalloprotease, was identified in the tropical liver fluke, Fasciola gigantica; that on sequence analysis showed a close homology (98.6%) with leucine aminopeptidase of the temperate liver fluke, Fasciola hepatica. The recombinant leucine aminopeptidase protein was expressed in Escherichia coli. F. gigantica peroxiredoxin, a hydrogen peroxide scavenger and an immunomodulating protein, was also cloned and expressed in E. coli. A vaccination trial in buffaloes was conducted with these two recombinant proteins, with 150 and 300μg of leucine aminopeptidase and a cocktail of 150μg each of recombinant leucine aminopeptidase and peroxiredoxin in three groups, respectively. Both Th1- and Th2-associated humoral immune responses were elicited to immunization with these antigens. A challenge study with 400 metacercariae did not show a significant protection in terms of reduction in the worm burden (8.4%) or anti-fecundity/embryonation effect in the immunized groups, as to the non-immunized control animals. Our observations in this buffalo vaccination trial are contrary to the earlier promise shown by leucine aminopeptidase of F. hepatica as a leading candidate vaccine molecule. Identification of leucine aminopeptidase gene and evaluation of the protein for its protective efficacy in buffaloes is the first scientific report on this protein in F. gigantica.
•Three recombinant antigens EgAgB8/1, EgAgB8/2 and EPC1 of hydatid cyst were expressed in prokaryotic expression vectors.•These three antigens showed fair degree of sensitivity but cross reacted with ...the parasites of buffalo host in IgG ELISA platform.•Sero-diagnosis of cystic echinococcosis in buffaloes is still a challenge.
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.
Fasciola gigantica
, causative agent of tropical fasciolosis
,
inflicts substantial economic losses on the livestock industry, affecting severely buffalo productivity in the tropical countries. Very ...few vaccination trials with different target antigens against
F. gigantica
infection have been conducted in this host. Present study describes a vaccination trial in buffaloes with
F. gigantica
recombinant glutathione S-transferase and fatty acid binding protein. The two recombinant proteins were expressed in
Escherichia coli
and evaluated for their immunoprophylactic potential in buffalo calves, using montanide 70 M-VG, a mineral oil-based adjuvant, for delivering the antigens. Buffalo calves were distributed in three groups, with group I, II and III calves immunized with recombinant glutathione S-transferase, fatty acid binding protein and a cocktail of these two antigens, respectively. Immunization of the calves evoked a mixed IgG1 and IgG2 antibody response. Present vaccination trial in these animals achieved a maximum protection level of 35%, when the two antigens were used in combination. Eosinophils were measured in both immunized and non-immunized challenge control animals, which showed a steady increase in their count in response to immunization with both the antigens and infection with
F. gigantica
, respectively.
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.
•Anaplasma marginale infection in carrier cattle from north-India detected by microscopy and nested-PCR.•A. marginale recombinant antigens MSP-2 and MSP-5 expressed in Escherichia coli.•Anaplasma specific antibodies detected in cattle by IgG and competitive inhibition ELISA.•msp5 nested-PCR highly sensitive and reliable for detection of A. marginale in carrier cattle.•A high prevalence of A. marginale infection detected in cattle in the region.
•A cDNA coding for Fasciola gigantica Cu/Zn–superoxide dismutase (SOD) was isolated.•Differential expression of the enzyme studied in the adult, juvenile and metacercaria.•Recombinant Cu/Zn–SOD ...partially characterized for enzyme activity, pH profile in NBT–PAGE.•Cu/Zn–SODs in the somatic and excretory secretory product of the fluke were studied.
Nitro blue tetrazolium–polyacrylamide gel (NBT–PAGE) showing Fasciola gigantica recombinant Cu/Zn–superoxide dismutase enzyme activity (lane 1) and effect of various concentrations of hydrogen peroxide (lanes 2, 3 and 4) and sodium azide (lanes 5, 6 and 7) on the Cu/Zn–SOD enzyme activity.
A full-length complementary DNA (cDNA) encoding Cu/Zn–superoxide dismutase was isolated from Fasciola gigantica that on nucleotide sequencing showed a close homology (98.9%) with Cu/Zn–superoxide dismutase (SOD) of the temperate liver fluke, F. hepatica. Expression of the gene was found in all the three developmental stages of the parasite viz. adult, newly excysted juvenile and metacercaria at transcriptional level by reverse transcription-polymerase chain reaction (RT-PCR) and at the protein level by Western blotting. F. gigantica Cu/Zn–SOD cDNA was cloned and expressed in Escherichia coli. Enzyme activity of the recombinant protein was determined by nitroblue tetrazolium (NBT)–polyacrylamide gel electrophoresis (PAGE) and this activity was inactivated by hydrogen peroxide but not by sodium azide, indicating that the recombinant protein is Cu/Zn–SOD. The enzyme activity was relatively stable at a broad pH range of pH 4.0–10.0. Native Cu/Zn–superoxide dismutase protein was detected in the somatic extract and excretory–secretory products of the adult F. gigantica by Western blotting. NBT–PAGE showed a single Cu/Zn–SOD present in the somatic extract while three SODs are released ex vivo by the adult parasite. The recombinant superoxide dismutase did not react with the serum from buffaloes infected with F. gigantica. The role of this enzyme in defense by the parasite against the host reactive oxygen species is discussed.
