Mrityunjay Mandal - Academia.edu (original) (raw)

Papers by Mrityunjay Mandal

Research paper thumbnail of Genetic diversity within ITS-1 region of Eimeria species infecting chickens of north India

Infection, Genetics and Evolution, 2015

Coccidiosis, caused by apicomplexan parasites of the genus Eimeria, inflicts severe economic loss... more Coccidiosis, caused by apicomplexan parasites of the genus Eimeria, inflicts severe economic losses to the poultry industry around the globe. In the present study, ITS-1 based species specific nested PCR revealed prevalence of E. acervulina, E. brunetti, E. maxima, E. mitis, E. praecox, E. necatrix and E. tenella in 79.2%, 12.5%, 64.6%, 89.6%, 60.4%, 64.6% and 97.9% poultry farms of north India, respectively. The ITS-1 sequences of different Eimeria spp. from north India were generated and analyzed to establish their phylogenetic relationship. The sequence identity with available sequences ranged from 80 to 100% in E. tenella, 95 to 100% in E. acervulina, 64 to 97% in E. necatrix, 96 to 99% in E. brunetti and 97 to 98% in E. mitis. Only long ITS-1 sequences of E. maxima could be generated in the present study and it had 80-100% identity with published sequences. Two out of the four ITS-1 sequences of E. maxima had mismatches in the published nested primer sequences from Australia, while one sequence of E. necatrix had a mismatch near 3′ end of both forward and reverse published nested primer sequences, warranting for the need of designing new set of degenerate primers for these two species of Eimeria. In the phylogenetic tree, all isolates of E. acervulina, E. brunetti, E. mitis, E. tenella and E. necatrix clustered in separate clades with high bootstrap value. E. maxima sequences of north Indian isolates grouped in a long form of E. maxima clade. Complete ITS-1 sequences of E. necatrix and E. mitis are reported for the first time from India. Further studies are required with more number of isolates to verify whether these differences are unique to geographical locations.

Research paper thumbnail of Development of loop-mediated isothermal amplification (LAMP) for detection of Babesia gibsoni infection in dogs

Veterinary Parasitology, Apr 15, 2015

Diagnosis of canine babesiosis, caused by Babesia gibsoni is difficult, especially in chronically... more Diagnosis of canine babesiosis, caused by Babesia gibsoni is difficult, especially in chronically infected dogs. A loop mediated isothermal amplification (LAMP) assay was developed and standardized by using four oligonucleotide primers targeting the hypervariable region of 18S rRNA gene (GenBank Acc. no. KC461261). The primers specifically amplified B. gibsoni DNA, while no amplification was detected with DNA from non-infected dogs as well as from dogs infected with Babesia canis vogeli, Hepatozoon canis, Ehrlichia canis and Trypanosoma evansi. The assay could detect 1.35×10(-7) parasitaemia and 10(-4) dilution of recombinant plasmid, equivalent to 12pg of target DNA. All the samples were tested by nested PCR as well as LAMP assay. LAMP was found to be 10 times more sensitive than nested PCR targeting the same gene. Out of 75 suspected field samples, collected from different parts of the country, LAMP could detect B. gibsoni in 43 samples, while nested PCR and microscopy could detect 37 and 23 samples, respectively. High sensitivity, specificity and rapidity of LAMP assay may be exploited for screening large number of samples in a field setting.

Research paper thumbnail of Histopathologial Alterations in Babesia Gibsoni Infectiona Case Report

Exploratory Animal and Medical Research, 2019

An eight-year-old male Labrador with high fever was brought for treatment in the Veterinary Clini... more An eight-year-old male Labrador with high fever was brought for treatment in the Veterinary Clinical Complex, Faculty of Veterinary and Animal Sciences, WBUAFS, Kolkata. Babesia gibsoni was identified in microscopical examination of the blood smear taken from the animal. The dog could not survive despite treatment and the post mortem examination revealed remarkable changes in the liver, lungs, kidneys and spleen. Histopathological examination showed congestion and edema in lungs, liver and spleen along with remarkable necrotic and degenerative changes in the tubular epithelial cell of kidneys.

Research paper thumbnail of Histopathological alterations in Babesia gibsoni infection - a case report

Exploratory Animal and Medical Research, 2019

Research paper thumbnail of Pathological changes in rats and rabbits experimentally infected with Trypanosoma evansi

Indian Journal of Veterinary Pathology, 2013

Macroscopical and microscopical lesions in visceral organs of experimental Trypanosoma evansi inf... more Macroscopical and microscopical lesions in visceral organs of experimental Trypanosoma evansi infected rats and New Zealand White rabbits were studied at the height of parasitaemia. Excessive accumulation of fluids in the abdominal and thoracic cavities, mottling with necrotic edges in liver, splenomegaly and congestion of meninges of brain were prominent gross lesions. Microscopically, fatty degeneration in liver, serous exudations under pleura, emphysema with infiltration of polymorphs, lymphocytes and mononuclear cells in lungs were found. The changes in the spleen include massive hyperplasia of lymphoid tissue at Malpighian corpuscles along with haemosiderin pigments. Kidneys showed hyaline degeneration and epithelial casts in the proximal and distal tubules. Congestion of meninges with focal submeningeal oedema with encephalomalacia was noticed in brain. It is proposed that the pathological changes of these organs might be due to immune complex deposition and complement cascade...

Research paper thumbnail of STUDIES ON OUU. ROUTE OF TRANSMISSION. SURVIVABILITY OF Trypanosoma evansi IN DEAD HOST AND PRESERVATIVE MEDIUM

Research paper thumbnail of Epidemiological studies for strategic control of gastrointestinal helminths in cattle population under Sub-Himalayan Terai Zone of West Bengal

The present study was aimed to understand the epidemiological prevalence of the gastrointestinal ... more The present study was aimed to understand the epidemiological prevalence of the gastrointestinal parasitism in cattle population. A total 1620 faecal samples were collected from three eco-regions under different seasonal conditions. Faecalysis was performed following standard protocol. GI helminths, nematode (Toxocara, Strongyloides, Strongyle and Trichuris), cestode (Moniezia) and trematode (Paramphistomes and Fasciola) were examined based on morphology of eggs. The overall prevalence was recorded upto 60.68% when Paramphistomes and Strongyle represented the highest and lowest degree of infection. The location specific study confirmed the highest magnitude of nematode, cestode and trematode infection at AlipurduarII, Falakata and Mathabhanga respectively. Compared to others, the rainy season was most congenial to GI parasitism. Young animals (6-24months) were severely affected with highest cestode infection (3.14%) among different age groups. This study affirms host, parasite and e...

Research paper thumbnail of PCR based identification of Babesia canis vogeli in clinically affected dogs

Canine babesiosis is a clinically important disease encountered around the globe. In India, the d... more Canine babesiosis is a clinically important disease encountered around the globe. In India, the disease, caused by large form of Babesia is thought to be exclusively due to B. canis, which is identified on the basis of microscopic examination of blood smears. It is now well accepted that B. canis of dogs across the world may look similar but vary in geographical distribution, tick vector preference, immunogenicity and pathogenicity. The present study was undertaken in three clinical cases of babesiosis from northern India to identify the parasite at subspecies level. PCR based amplification of target DNA using specific primers for different subspecies of B. canis revealed the presence of B. canis vogeli in all the three cases.

Research paper thumbnail of Oral route of transmission: Trypanosoma evansi in a mice model experiment

Journal of Parasitic Diseases

Twelve Swiss albino mice of either sex and equal body weight were randomly divided in 2 groups (I... more Twelve Swiss albino mice of either sex and equal body weight were randomly divided in 2 groups (I and II), consisting of 9 and 3 mice respectively and were used to conduct the study. A dose of 2.5 × 104 number of Trypanosoma evansi was instantly fed to each mouse of group I. Each mouse of group II was inoculated intraperitoneally with same dose of parasites through infected mice blood and kept separate. The tail blood of each mouse was examined daily up to 30 days post infection by examination of wet blood film and Giemsa-stained blood smears for presence of any trypanosomes. Out of 9 mice of group I those were infected orally, 3 (33.33%) mice became positive for presence of T. evansi both by examination of wet blood film and Giemsa-stained blood smears after 4, 6 and 7 days post infection. After 2 days post infection all intraperitoneally infected mice were found positive for T. evansi. Thus incubation period in orally infected mice was longer than the intraperitoneally infected mice. All the positive mice of both the groups died with high parasitaemia after 3–4 days of first appearance of parasitaemia. From the present study, it can be concluded that besides mechanical or parenteral means of transmission, T. evansi could also be transmitted through oral route. Thus zoo carnivores might be infected with T. evansi and develop disease by eating infected blood or flesh of the infected animals, as a prey and predator relationship.

Research paper thumbnail of Humoral and cytokine response elicited during immunisation with recombinant Immune Mapped protein-1 (EtIMP-1) and oocysts of Eimeria tenella

Research paper thumbnail of Bovine neosporosis: an overview

North East Veterinarian, 2011

Research paper thumbnail of Integrated parasite management for livestock - alternative control methods

Veterinary World, 2010

Internal parasites are considered by some to be one of the most economically important constraint... more Internal parasites are considered by some to be one of the most economically important constraints in raising livestock. The growing concern about the resistance of internal parasites to all classes of dewormers has caused people to look for alternatives. As dewormers lose their effectiveness, the livestock community fears increasing economic losses from worms. There is no one thing that can be given or done to replace chemical dewormers. It will take a combination of extremely good management techniques and possibly some alternative therapies. It is not wise to think that one can just stop deworming animals with chemical dewormers. It is something one will need to change gradually, observing and testing animals and soil, in order to monitor the progress. Alternative parasite control is an area that is receiving a lot of interest and attention. Programs and research will continue in the pursuit of parasite control, using alternative and more management-intensive methods.

Research paper thumbnail of Development of recombinant BgP12 based enzyme linked immunosorbent assays for serodiagnosis of Babesia gibsoni infection in dogs

Veterinary Immunology and Immunopathology, 2015

Indirect ELISA and dot-ELISA using recombinant BgP12 (rBgP12) were developed for the diagnosis of... more Indirect ELISA and dot-ELISA using recombinant BgP12 (rBgP12) were developed for the diagnosis of Babesia gibsoni infected dogs. The complete open reading frame of BgP12 gene (378bp) was cloned in pET-32a(+) expression vector and expressed in Escherichia coli as a soluble thioredoxin (Trx) fusion protein. The purified rBgP12 was used for production of anti-rBgP12 rabbit serum, which recognized a native 12-kDa protein in B. gibsoni infected erythrocyte by Western blot analysis. To evaluate the potential of rBgP12 for the serodiagnosis of B. gibsoni, a panel of serum/plasma samples from dogs infected with B. gibsoni (n=13), uninfected sera (n=13) and sera from dogs infected with other haemoparasites viz., Babesia canis vogeli (n=3), Ehrlichia canis (n=3), Hepatozoon canis (n=1) and Dirofilaria immitis (n=1) were used in ELISA formats. In addition, the performance of rBgP12 based indirect ELISA and dot-ELISA were evaluated using 75 serum/plasma samples collected from suspected dogs, in respect to the nested PCR as reference test. The diagnostic sensitivities of indirect ELISA and dot-ELISA were 94.59% and 89.18%, respectively, while their specificities were 84.21% and 81.57%, respectively. Moreover, both the assays using rBgP12 showed no cross reaction with sera from dogs infected with other common haemoparasites indicating their high specificity. High kappa values of indirect ELISA and dot-ELISA indicated the potentials of these assays with substantial agreement at 95% confidence level. It is concluded that indirect ELISA and dot ELISA using rBgP12 might be used in large scale epidemiological surveys and clinical diagnosis of B. gibsoni infection in dogs.

Research paper thumbnail of Experimental studies on survivality and degenerative changes of Trypanosoma evansi after death of host

Journal of Parasitic Diseases, 2013

Twenty adult Swiss albino mice, 20 rats and 10 rabbits were artificially infected with Trypanosom... more Twenty adult Swiss albino mice, 20 rats and 10 rabbits were artificially infected with Trypanosoma evansi and killed at the peak of parasitaemia to know the period of survivality of T. evansi and degenerative changes of the parasite after death of these hosts. Examination of Giemsa stained blood smears and wet blood smears revealed the presence of parasites and live trypanosomes along with motility in the heart blood of mice and rats up to 14 h and in rabbits up to 13 h post death. Mouse inoculation test (MIT) conducted with heart blood up to 13 h post death of mice and rabbits became positive. MIT with both heart blood and portal blood of rats became positive up to 14 h post death. The liver and lung impression smears could detect the parasites up to 14 h of death of mice and rats and up to 13 h post death of rabbits whereas spleen impression smears revealed the presence of parasites up to 12 h post death of these animals. It is confirmed that T. evansi infection in animals may be diagnosed after post mortem examination of hosts by demonstration of parasites.

Research paper thumbnail of Development of loop-mediated isothermal amplification (LAMP) for detection of Babesia gibsoni infection in dogs

Veterinary Parasitology, 2015

Research paper thumbnail of Genetic characterization and phylogenetic relationships based on 18S rRNA and ITS1 region of small form of canine Babesia spp. from India

Infection, Genetics and Evolution, 2014

Research paper thumbnail of Cloning and sequencing of beta-tubulin and internal transcribed spacer-2 (ITS-2) of Eimeria tenella isolate from India

Journal of Parasitic Diseases, 2013

Research paper thumbnail of Development and evaluation of serodiagnostic assays with recombinant BgSA1 of Babesia gibsoni

Veterinary Parasitology, 2014

Research paper thumbnail of Development of a Loop-Mediated Isothermal Amplification Assay for Rapid, Sensitive and Specific Detection of a Campylobacter jejuni Clone

Loop-mediated isothermal amplification (LAMP) assay is a simple, rapid and specific detection met... more Loop-mediated isothermal amplification (LAMP) assay is a simple, rapid and specific detection method and has been used for detection and identification of different Campylobacter species. In this study, we develop a LAMP assay specific for detection of a particular clone (clone SA) of Campylobacter jejuni, associated with the vast majority of recent sheep abortions in the U.S. Using a set of specific primers for C. jejuni IA3902 (a clone SA isolate) and genomic DNA or boiled cell extract as template, the target DNA was amplified at 63℃ for 50 min in a water bath. A positive reaction was identified visually as white precipitate or fluorescence under UV, and confirmed by gel electrophoresis. Detection limit of the assay was comparable to that of conventional PCR. The LAMP was shown to be specific for detection of clone SA when tested on a number of C. jejuni strains of different genetic backgrounds. Applicability of the LAMP assay for specific detection of clone SA was demonstrated in...

Research paper thumbnail of Development of loop-mediated isothermal amplification (LAMP) for detection of Babesia gibsoni infection in dogs

Veterinary parasitology, Jan 19, 2015

Diagnosis of canine babesiosis, caused by Babesia gibsoni is difficult, especially in chronically... more Diagnosis of canine babesiosis, caused by Babesia gibsoni is difficult, especially in chronically infected dogs. A loop mediated isothermal amplification (LAMP) assay was developed and standardized by using four oligonucleotide primers targeting the hypervariable region of 18S rRNA gene (GenBank Acc. no. KC461261). The primers specifically amplified B. gibsoni DNA, while no amplification was detected with DNA from non-infected dogs as well as from dogs infected with Babesia canis vogeli, Hepatozoon canis, Ehrlichia canis and Trypanosoma evansi. The assay could detect 1.35×10(-7) parasitaemia and 10(-4) dilution of recombinant plasmid, equivalent to 12pg of target DNA. All the samples were tested by nested PCR as well as LAMP assay. LAMP was found to be 10 times more sensitive than nested PCR targeting the same gene. Out of 75 suspected field samples, collected from different parts of the country, LAMP could detect B. gibsoni in 43 samples, while nested PCR and microscopy could detec...

Research paper thumbnail of Genetic diversity within ITS-1 region of Eimeria species infecting chickens of north India

Infection, Genetics and Evolution, 2015

Coccidiosis, caused by apicomplexan parasites of the genus Eimeria, inflicts severe economic loss... more Coccidiosis, caused by apicomplexan parasites of the genus Eimeria, inflicts severe economic losses to the poultry industry around the globe. In the present study, ITS-1 based species specific nested PCR revealed prevalence of E. acervulina, E. brunetti, E. maxima, E. mitis, E. praecox, E. necatrix and E. tenella in 79.2%, 12.5%, 64.6%, 89.6%, 60.4%, 64.6% and 97.9% poultry farms of north India, respectively. The ITS-1 sequences of different Eimeria spp. from north India were generated and analyzed to establish their phylogenetic relationship. The sequence identity with available sequences ranged from 80 to 100% in E. tenella, 95 to 100% in E. acervulina, 64 to 97% in E. necatrix, 96 to 99% in E. brunetti and 97 to 98% in E. mitis. Only long ITS-1 sequences of E. maxima could be generated in the present study and it had 80-100% identity with published sequences. Two out of the four ITS-1 sequences of E. maxima had mismatches in the published nested primer sequences from Australia, while one sequence of E. necatrix had a mismatch near 3′ end of both forward and reverse published nested primer sequences, warranting for the need of designing new set of degenerate primers for these two species of Eimeria. In the phylogenetic tree, all isolates of E. acervulina, E. brunetti, E. mitis, E. tenella and E. necatrix clustered in separate clades with high bootstrap value. E. maxima sequences of north Indian isolates grouped in a long form of E. maxima clade. Complete ITS-1 sequences of E. necatrix and E. mitis are reported for the first time from India. Further studies are required with more number of isolates to verify whether these differences are unique to geographical locations.

Research paper thumbnail of Development of loop-mediated isothermal amplification (LAMP) for detection of Babesia gibsoni infection in dogs

Veterinary Parasitology, Apr 15, 2015

Diagnosis of canine babesiosis, caused by Babesia gibsoni is difficult, especially in chronically... more Diagnosis of canine babesiosis, caused by Babesia gibsoni is difficult, especially in chronically infected dogs. A loop mediated isothermal amplification (LAMP) assay was developed and standardized by using four oligonucleotide primers targeting the hypervariable region of 18S rRNA gene (GenBank Acc. no. KC461261). The primers specifically amplified B. gibsoni DNA, while no amplification was detected with DNA from non-infected dogs as well as from dogs infected with Babesia canis vogeli, Hepatozoon canis, Ehrlichia canis and Trypanosoma evansi. The assay could detect 1.35×10(-7) parasitaemia and 10(-4) dilution of recombinant plasmid, equivalent to 12pg of target DNA. All the samples were tested by nested PCR as well as LAMP assay. LAMP was found to be 10 times more sensitive than nested PCR targeting the same gene. Out of 75 suspected field samples, collected from different parts of the country, LAMP could detect B. gibsoni in 43 samples, while nested PCR and microscopy could detect 37 and 23 samples, respectively. High sensitivity, specificity and rapidity of LAMP assay may be exploited for screening large number of samples in a field setting.

Research paper thumbnail of Histopathologial Alterations in Babesia Gibsoni Infectiona Case Report

Exploratory Animal and Medical Research, 2019

An eight-year-old male Labrador with high fever was brought for treatment in the Veterinary Clini... more An eight-year-old male Labrador with high fever was brought for treatment in the Veterinary Clinical Complex, Faculty of Veterinary and Animal Sciences, WBUAFS, Kolkata. Babesia gibsoni was identified in microscopical examination of the blood smear taken from the animal. The dog could not survive despite treatment and the post mortem examination revealed remarkable changes in the liver, lungs, kidneys and spleen. Histopathological examination showed congestion and edema in lungs, liver and spleen along with remarkable necrotic and degenerative changes in the tubular epithelial cell of kidneys.

Research paper thumbnail of Histopathological alterations in Babesia gibsoni infection - a case report

Exploratory Animal and Medical Research, 2019

Research paper thumbnail of Pathological changes in rats and rabbits experimentally infected with Trypanosoma evansi

Indian Journal of Veterinary Pathology, 2013

Macroscopical and microscopical lesions in visceral organs of experimental Trypanosoma evansi inf... more Macroscopical and microscopical lesions in visceral organs of experimental Trypanosoma evansi infected rats and New Zealand White rabbits were studied at the height of parasitaemia. Excessive accumulation of fluids in the abdominal and thoracic cavities, mottling with necrotic edges in liver, splenomegaly and congestion of meninges of brain were prominent gross lesions. Microscopically, fatty degeneration in liver, serous exudations under pleura, emphysema with infiltration of polymorphs, lymphocytes and mononuclear cells in lungs were found. The changes in the spleen include massive hyperplasia of lymphoid tissue at Malpighian corpuscles along with haemosiderin pigments. Kidneys showed hyaline degeneration and epithelial casts in the proximal and distal tubules. Congestion of meninges with focal submeningeal oedema with encephalomalacia was noticed in brain. It is proposed that the pathological changes of these organs might be due to immune complex deposition and complement cascade...

Research paper thumbnail of STUDIES ON OUU. ROUTE OF TRANSMISSION. SURVIVABILITY OF Trypanosoma evansi IN DEAD HOST AND PRESERVATIVE MEDIUM

Research paper thumbnail of Epidemiological studies for strategic control of gastrointestinal helminths in cattle population under Sub-Himalayan Terai Zone of West Bengal

The present study was aimed to understand the epidemiological prevalence of the gastrointestinal ... more The present study was aimed to understand the epidemiological prevalence of the gastrointestinal parasitism in cattle population. A total 1620 faecal samples were collected from three eco-regions under different seasonal conditions. Faecalysis was performed following standard protocol. GI helminths, nematode (Toxocara, Strongyloides, Strongyle and Trichuris), cestode (Moniezia) and trematode (Paramphistomes and Fasciola) were examined based on morphology of eggs. The overall prevalence was recorded upto 60.68% when Paramphistomes and Strongyle represented the highest and lowest degree of infection. The location specific study confirmed the highest magnitude of nematode, cestode and trematode infection at AlipurduarII, Falakata and Mathabhanga respectively. Compared to others, the rainy season was most congenial to GI parasitism. Young animals (6-24months) were severely affected with highest cestode infection (3.14%) among different age groups. This study affirms host, parasite and e...

Research paper thumbnail of PCR based identification of Babesia canis vogeli in clinically affected dogs

Canine babesiosis is a clinically important disease encountered around the globe. In India, the d... more Canine babesiosis is a clinically important disease encountered around the globe. In India, the disease, caused by large form of Babesia is thought to be exclusively due to B. canis, which is identified on the basis of microscopic examination of blood smears. It is now well accepted that B. canis of dogs across the world may look similar but vary in geographical distribution, tick vector preference, immunogenicity and pathogenicity. The present study was undertaken in three clinical cases of babesiosis from northern India to identify the parasite at subspecies level. PCR based amplification of target DNA using specific primers for different subspecies of B. canis revealed the presence of B. canis vogeli in all the three cases.

Research paper thumbnail of Oral route of transmission: Trypanosoma evansi in a mice model experiment

Journal of Parasitic Diseases

Twelve Swiss albino mice of either sex and equal body weight were randomly divided in 2 groups (I... more Twelve Swiss albino mice of either sex and equal body weight were randomly divided in 2 groups (I and II), consisting of 9 and 3 mice respectively and were used to conduct the study. A dose of 2.5 × 104 number of Trypanosoma evansi was instantly fed to each mouse of group I. Each mouse of group II was inoculated intraperitoneally with same dose of parasites through infected mice blood and kept separate. The tail blood of each mouse was examined daily up to 30 days post infection by examination of wet blood film and Giemsa-stained blood smears for presence of any trypanosomes. Out of 9 mice of group I those were infected orally, 3 (33.33%) mice became positive for presence of T. evansi both by examination of wet blood film and Giemsa-stained blood smears after 4, 6 and 7 days post infection. After 2 days post infection all intraperitoneally infected mice were found positive for T. evansi. Thus incubation period in orally infected mice was longer than the intraperitoneally infected mice. All the positive mice of both the groups died with high parasitaemia after 3–4 days of first appearance of parasitaemia. From the present study, it can be concluded that besides mechanical or parenteral means of transmission, T. evansi could also be transmitted through oral route. Thus zoo carnivores might be infected with T. evansi and develop disease by eating infected blood or flesh of the infected animals, as a prey and predator relationship.

Research paper thumbnail of Humoral and cytokine response elicited during immunisation with recombinant Immune Mapped protein-1 (EtIMP-1) and oocysts of Eimeria tenella

Research paper thumbnail of Bovine neosporosis: an overview

North East Veterinarian, 2011

Research paper thumbnail of Integrated parasite management for livestock - alternative control methods

Veterinary World, 2010

Internal parasites are considered by some to be one of the most economically important constraint... more Internal parasites are considered by some to be one of the most economically important constraints in raising livestock. The growing concern about the resistance of internal parasites to all classes of dewormers has caused people to look for alternatives. As dewormers lose their effectiveness, the livestock community fears increasing economic losses from worms. There is no one thing that can be given or done to replace chemical dewormers. It will take a combination of extremely good management techniques and possibly some alternative therapies. It is not wise to think that one can just stop deworming animals with chemical dewormers. It is something one will need to change gradually, observing and testing animals and soil, in order to monitor the progress. Alternative parasite control is an area that is receiving a lot of interest and attention. Programs and research will continue in the pursuit of parasite control, using alternative and more management-intensive methods.

Research paper thumbnail of Development of recombinant BgP12 based enzyme linked immunosorbent assays for serodiagnosis of Babesia gibsoni infection in dogs

Veterinary Immunology and Immunopathology, 2015

Indirect ELISA and dot-ELISA using recombinant BgP12 (rBgP12) were developed for the diagnosis of... more Indirect ELISA and dot-ELISA using recombinant BgP12 (rBgP12) were developed for the diagnosis of Babesia gibsoni infected dogs. The complete open reading frame of BgP12 gene (378bp) was cloned in pET-32a(+) expression vector and expressed in Escherichia coli as a soluble thioredoxin (Trx) fusion protein. The purified rBgP12 was used for production of anti-rBgP12 rabbit serum, which recognized a native 12-kDa protein in B. gibsoni infected erythrocyte by Western blot analysis. To evaluate the potential of rBgP12 for the serodiagnosis of B. gibsoni, a panel of serum/plasma samples from dogs infected with B. gibsoni (n=13), uninfected sera (n=13) and sera from dogs infected with other haemoparasites viz., Babesia canis vogeli (n=3), Ehrlichia canis (n=3), Hepatozoon canis (n=1) and Dirofilaria immitis (n=1) were used in ELISA formats. In addition, the performance of rBgP12 based indirect ELISA and dot-ELISA were evaluated using 75 serum/plasma samples collected from suspected dogs, in respect to the nested PCR as reference test. The diagnostic sensitivities of indirect ELISA and dot-ELISA were 94.59% and 89.18%, respectively, while their specificities were 84.21% and 81.57%, respectively. Moreover, both the assays using rBgP12 showed no cross reaction with sera from dogs infected with other common haemoparasites indicating their high specificity. High kappa values of indirect ELISA and dot-ELISA indicated the potentials of these assays with substantial agreement at 95% confidence level. It is concluded that indirect ELISA and dot ELISA using rBgP12 might be used in large scale epidemiological surveys and clinical diagnosis of B. gibsoni infection in dogs.

Research paper thumbnail of Experimental studies on survivality and degenerative changes of Trypanosoma evansi after death of host

Journal of Parasitic Diseases, 2013

Twenty adult Swiss albino mice, 20 rats and 10 rabbits were artificially infected with Trypanosom... more Twenty adult Swiss albino mice, 20 rats and 10 rabbits were artificially infected with Trypanosoma evansi and killed at the peak of parasitaemia to know the period of survivality of T. evansi and degenerative changes of the parasite after death of these hosts. Examination of Giemsa stained blood smears and wet blood smears revealed the presence of parasites and live trypanosomes along with motility in the heart blood of mice and rats up to 14 h and in rabbits up to 13 h post death. Mouse inoculation test (MIT) conducted with heart blood up to 13 h post death of mice and rabbits became positive. MIT with both heart blood and portal blood of rats became positive up to 14 h post death. The liver and lung impression smears could detect the parasites up to 14 h of death of mice and rats and up to 13 h post death of rabbits whereas spleen impression smears revealed the presence of parasites up to 12 h post death of these animals. It is confirmed that T. evansi infection in animals may be diagnosed after post mortem examination of hosts by demonstration of parasites.

Research paper thumbnail of Development of loop-mediated isothermal amplification (LAMP) for detection of Babesia gibsoni infection in dogs

Veterinary Parasitology, 2015

Research paper thumbnail of Genetic characterization and phylogenetic relationships based on 18S rRNA and ITS1 region of small form of canine Babesia spp. from India

Infection, Genetics and Evolution, 2014

Research paper thumbnail of Cloning and sequencing of beta-tubulin and internal transcribed spacer-2 (ITS-2) of Eimeria tenella isolate from India

Journal of Parasitic Diseases, 2013

Research paper thumbnail of Development and evaluation of serodiagnostic assays with recombinant BgSA1 of Babesia gibsoni

Veterinary Parasitology, 2014

Research paper thumbnail of Development of a Loop-Mediated Isothermal Amplification Assay for Rapid, Sensitive and Specific Detection of a Campylobacter jejuni Clone

Loop-mediated isothermal amplification (LAMP) assay is a simple, rapid and specific detection met... more Loop-mediated isothermal amplification (LAMP) assay is a simple, rapid and specific detection method and has been used for detection and identification of different Campylobacter species. In this study, we develop a LAMP assay specific for detection of a particular clone (clone SA) of Campylobacter jejuni, associated with the vast majority of recent sheep abortions in the U.S. Using a set of specific primers for C. jejuni IA3902 (a clone SA isolate) and genomic DNA or boiled cell extract as template, the target DNA was amplified at 63℃ for 50 min in a water bath. A positive reaction was identified visually as white precipitate or fluorescence under UV, and confirmed by gel electrophoresis. Detection limit of the assay was comparable to that of conventional PCR. The LAMP was shown to be specific for detection of clone SA when tested on a number of C. jejuni strains of different genetic backgrounds. Applicability of the LAMP assay for specific detection of clone SA was demonstrated in...

Research paper thumbnail of Development of loop-mediated isothermal amplification (LAMP) for detection of Babesia gibsoni infection in dogs

Veterinary parasitology, Jan 19, 2015

Diagnosis of canine babesiosis, caused by Babesia gibsoni is difficult, especially in chronically... more Diagnosis of canine babesiosis, caused by Babesia gibsoni is difficult, especially in chronically infected dogs. A loop mediated isothermal amplification (LAMP) assay was developed and standardized by using four oligonucleotide primers targeting the hypervariable region of 18S rRNA gene (GenBank Acc. no. KC461261). The primers specifically amplified B. gibsoni DNA, while no amplification was detected with DNA from non-infected dogs as well as from dogs infected with Babesia canis vogeli, Hepatozoon canis, Ehrlichia canis and Trypanosoma evansi. The assay could detect 1.35×10(-7) parasitaemia and 10(-4) dilution of recombinant plasmid, equivalent to 12pg of target DNA. All the samples were tested by nested PCR as well as LAMP assay. LAMP was found to be 10 times more sensitive than nested PCR targeting the same gene. Out of 75 suspected field samples, collected from different parts of the country, LAMP could detect B. gibsoni in 43 samples, while nested PCR and microscopy could detec...