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Subject: Veterinary Parasitology ×

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    ThesisItemOpen Access
    STUDIES ON VECTOR POTENTIALITY OF IXODID TICKS OF CATTLE AND THEIR CONTROL
    (KARNATAKA VETERINARY, ANIMAL AND FISHERIES SCIENCES UNIVERSITY, BIDAR - 585 401, 2022) YUVARAJA, T. Y.; ANANDA, K. J.
    A systematic study on tick infestation in cattle from Kolar, Ramanagra, Chikkaballapura districts of Karnataka was carried out to know the prevalence status, morphological and molecular identification of ticks, evaluation of acaricidal activity of phytotherapeutic compounds and to study the vector potentiality of ixodid ticks of cattle by molecular method. The prevalence of 55.3 per cent tick infestation was recorded in cattle and district wise prevalence revealed 56, 52 and 58 per cent in Kolar, Ramanagara and Chikkaballapura districts of Karnataka respectively. Morphologically, the ticks were identified as Rhipicephalus microplus, Rhipicephalus haemaphysaloides, Hyalomma spp. and PCR targeting 12S rRNA gene revealed three Rhipicephalus microplus, Rhipicephalus haemaphysaloides and Haemaphysalis longicornis by sequence and phylogenetic analysis. Acaricidal activity of deltamethrin, piperine, nanopiperine, curcumin and nanocurcumin was done by larval packet test (LPT) and adult immersion test (AIT). LPT against R. microplus larvae showed an LC50 at 3.292 ppm for nanopiperine followed by 3.5 ppm, 5.8 ppm, 5.9 ppm and 73.435 ppm for nanocurcumin, curcumin, piperine, and deltamethrin respectively. AIT against R. microplus showed an LC50 at concentration of 5.652 for nanopiperine followed by 8.090 for nanocurcumin, 26.489 for piperine, 32.581 for curcumin and 106.594 for deltamethrin. In both LPT and AIT, LC50 values of nanopiperine and nanocurcumin were significantly lower compared to deltamethrin. Nanopiperine and nanocurcumin showed significant ovulation inhibitory activity with lower IC50 and IC99 values compared to piperine, curcumin and deltamethrin. The vector potentiality of cattle tick was studied by molecular method in 50 engorged ticks DNA randomly selected from 600 pooled tick samples collected from 120 cattle. The PCR was carried out by targeting MPSP, 18S rRNA, RpoB and GroEL genes of Theileria, Babesia, Anaplasma 171 and Ehrlichia species using specific primers revealed highest prevalence of Theileria oriantalis (33/66 %) followed by Babesia spp. (19/38 %), Anaplasma marginale (9/18 %), Ehrlichia minasensis (6/12 %) and Theileria annulata (2/4 %) whereas, 21 (42 %) showed positive for mixed infection. Thus, the study provides key information on the distribution of ticks and tick-borne pathogens of cattle in three districts of Karnataka, which will aid in developing and strategizing effective control measures. Keywords: Cattle, Ticks, Acaricidal activity, Vector potentiality, PCR
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    ThesisItemOpen Access
    STUDIES ON IXODID TICKS OF DOG WITH SPECIAL REFERENCE TO VECTOR POTENTIALITY BY MOLECULAR METHOD
    (KARNATAKA VETERINARY, ANIMAL AND FISHERIES SCIENCES UNIVERSITY, BIDAR - 585 401, 2022) SUNITHA, C. R.; ANANDA. K. J.
    A systematic study on tick infestation of dogs presented to the Veterinary college hospital, Hebbal, breeders, pet clinics and non-governmental organisations located in and around Bengaluru was carried out to know the prevalence status, morphological and molecular identification of ticks, evaluation of acaricidal activity of phytotherapeutic compounds and vector potentiality of ixodid ticks of dog by molecular method. The overall prevalence of tick infestation in dogs was recorded as 44 per cent in and around Bengaluru. Morphologically, the species of ticks were identified as Rhipicephalus sanguineus, Rhipicephalus haemaphysaloides and Haemaphysalis sp. and PCR targeting 12S rRNA gene revealed three tick species such as, Rhipicephalus sanguineus, Rhipicephalus haemaphysaloides and Haemaphysalis longicornis by sequence and phylogenetic analysis. Acaricidal activity of deltamethrin, piperine, nanopiperine, curcumin and nanocurcumin was done by larval packet test (LPT) and adult immersion test (AIT). LPT against R. sanguineus larvae showed an LC50 at 2.89 ppm for nanopiperine followed by 6.2 ppm, 6.95 ppm, 32.5 ppm and 64.05 ppm for piperine, nanocurcumin, curcumin and deltamethrin respectively. AIT against R. sanguineus showed an LC50 at concentration of 5.652 for nanopiperine followed by 6.959 for nanocurcumin, 19.863 for piperine, 32.581 for curcumin and 161.017 for deltamethrin. In both LPT and AIT, LC50 values of nanopiperine and nanocurcumin were significantly lower compared to deltamethrin. Nanopiperine and nanocurcumin showed significant ovulation inhibitory activity with lower IC50 and IC99 values compared to piperine, curcumin and deltamethrin. The vector potentiality of dog tick was studied by molecular method in 50 ticks DNA randomly selected from 600 pooled tick samples. PCR was carried out by targeting GroEL, 18S rRNA, 16S rRNA and RpoBgenes of Ehrlichia, Babesia and Hepatozoon, Haemobortenella and Anaplasma 146 species respectively using specific primers. The results of the molecular study showed highest prevalence of Ehrlichia canis (13/26 %) followed by Babesia spp. (7/14 %), Hepatozoon canis (5/10%) and Mycoplasma haemocanis (5/10 %) along with 6 (12 %) mixed infection. However, none of the tick DNA sample revealed positive by PCR targeting RpoB genes of Anaplasma species. Thus, the study provides key information on the distribution of ticks and tick-borne pathogens of dogs in and around Bengaluru, which will aid in the developing and strategizing effective control measures. Keywords: Dog, Ticks, Acaricidal activity, Vector potentiality, PCR
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    ThesisItemOpen Access
    DEVELOPMENT OF DRY LAMP AS POINT OF CARE DIAGNOSTICS FOR THE DIAGNOSIS OF BOVINE THEILERIOSIS
    (KARNATAKA VETERINARY, ANIMAL AND FISHERIES SCIENCES UNIVERSITY, BIDAR - 585 401, 2022) SINDHOORA DIVAKAR; DHANALAKSHMI. H.)
    Tropical theilerosis is an economically important vector-borne disease of tropical and subtropical parts of the world including India. Though there are many advancements being made for the diagnosis of theileriosis, they could not be efficiently applied to the field level. Hence, a study was undertaken to develop a dry LAMP assay as point of care diagnostics of diagnosis of theileriosis in cattle. Both wet and dry LAMP assays targeting cytochrome b gene were standardized for diagnosis of Theileria annulata in cattle and comparative evaluation was done. To state in terms of sensitivity it was found that both wet LAMP and dry LAMP could amplify as least as 10-4ng (0.1pg) of DNA. In terms of specificity it was found that both wet and dry LAMP assay amplified only Theileria annulata and did not amplify other closely related haemoprotozoans. A total of 200 suspected blood samples were subjected to microscopy, PCR , wet LAMP and dry LAMP assay. It was found that 16 samples were found to be positive for theilerosis in microscopy, 20 samples were found to be positive in PCR and 23 samples were found to be positive in both wet and dry LAMP assay. Hence, we can conclude that the LAMP assay (dry and wet LAMP) was found to be more sensitive (100%) when compared to microscopy (69.5%) and PCR (86.9%). In terms of shelf life, it was observed that the dry LAMP reaction tubes at room temperature as well as refrigeration temperature provided successful amplification till 7 weeks. Hence it can be concluded that using dry LAMP assay, one can develop a point of care diagnostic kit for the field level diagnosis of T.annulata in cattle.
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    ThesisItemOpen Access
    DEVELOPMENT OF LOOP MEDIATED ISOTHERMAL AMPLIFICATION (LAMP) ASSAY FOR THE DIAGNOSIS OF THEILERIOSIS IN CATTLE
    (KARNATAKA VETERINARY, ANIMAL AND FISHERIES SCIENCES UNIVERSITY, BIDAR - 585 401, 2022) SANDEEP. N; DHANALAKSHMI, H.)
    A study on 200 blood samples collected from different parts of Karnataka was undertaken to develop a paper based LAMP assay and a foldable isothermal amplification micro-device for the diagnosis of theileriosis in cattle. Both HNB wet LAMP and phenol red wet LAMP assays were standardized for diagnosis of Theileria annulata in cattle. Half the volume of optimized HNB wet LAMP was dried on paper substrate for the standardization of paper HNB LAMP with modification in dye concentration and this produced a color change from violet to blue. Whereas the same volume of phenol red wet LAMP was unable to produce desired color change on paper due to inherent paper character. And same standardized HNB paper LAMP was used for the standardization of foldable isothermal amplification micro-device and this produced a color change from initial violet to colorless. We fabricated and standardized a simple, cost-effective, compact and easy to operate paper LAMP and foldable isothermal amplification microdevice which can be employed for POCT and comparative evaluation was done. To state in terms of sensitivity it was found that both wet LAMP assays and paper LAMP assays could amplify as least as 10-4ng (0.1pg) of DNA. To state in terms of specificity it was found that both wet and paper LAMP assays amplified only Theileria annulata and did not amplify other closely related haemoprotozoans. All the 200 suspected blood samples were subjected to microscopy, PCR, wet LAMP assays and paper LAMP assays. It was found that 16 samples were found to be positive for theilerosis in microscopy, 20 samples were found to be positive in PCR and 23 samples were found positive in both wet and paper LAMP assays. Hence we can say that the LAMP assays (wet and paper LAMP) was found to be more sensitive (100%) when compared to microscopy (69.5%) and PCR (86.9%). The drying of LAMP reagents on paper substrate enables us to transport LAMP kits to areas where the cold chain is not easily available and employed for POCT because its single step operation.
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    ThesisItemOpen Access
    EFFICACY OF ANTITRYPANOSOMAL DRUGS AGAINST TRYPANOSOMA EVANSI BY IN VIVO STUDIES
    (KARNATAKA VETERINARY, ANIMAL AND FISHERIES SCIENCES UNIVERSITY, BIDAR- 585 401, 2022) NAGARAJ JALLER; JAYA NAGAPPA LAKKUNDI
    Efficacy of aqueous fresh leaf extract and silver nanoparticles of L. camara (LCAgNPs) along with quinapyramine prosalts were evaluated by in vivo studies in mice model against T. evansi (buffalo isolate) in Karnataka state, India. Ten out of 11 experimental mice groups consisting 8 in each were inoculated with 5x104 T. evansi and one group was maintained as negative control. The infected mice showed parasitaemia on 2nd day post inoculum. One infected group was treated subcutaneously with quinapyramine prosalts at 5 mg/kg b.w. Four infected groups treated with aqueous fresh leaf extract and LCAgNPs at 0.417 and 0.208 g/kg b.w intraperitoneally for three consecutive days. Similarly, four groups treated in combination with quinapyramine prosalts. Two mice from each group sacrificed on 8, 15, 28 and 45th day post treatment for histopathology and PCR analysis. On 5th day 100.0 % mortality observed in positive, aqueous fresh leaf extract and LCAgNPs treated groups and showed congested lungs, hepatomegaly and splenomegaly with major histopathological changes along with organisms in blood vessels of vital organs. Whereas, 12.5 % mortality observed on 30th day in quinapyramine treated group due to relapse and no mortality in other groups with absence of organisms from 2 to 45 days post treatment showed no significant difference between quinapyramine prosalts treated and combination of drug, LCAgNPS and aqueous fresh leaf extract treated groups. PCR analysis of blood of sacrificed mice revealed absence of relapse. The present study concluded post treatment evaluation of quinapyramine prosalts for relapse and further exploration of phytochemicals against trypanosomosis.
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    ThesisItemOpen Access
    SYNTHESIS AND EVALUATION OF PLANT BASED SILVER NANOPARTICLES ON MUSCID FLIES
    (KARNATAKA VETERINARY, ANIMAL AND FISHERIES SCIENCES UNIVERSITY, BIDAR- 585 401, 2022) JAGADISH; MAMATHA G. S.)
    An insecticidal efficacy of AgNPs synthesized from aqueous leaf extracts of E. tereticornis (Et-AgNPs) and P. pinnata (Pp-AgNPs) was evaluated on muscid flies by in vitro assays at different concentrations. The highest mortality of 70.0 per cent was observed against the adult flies of S. calcitrans and M. domestica in Et and Pp-AgNPs, at 20 mg concentration after 24 hours exposure. The LC25(1.226, 1.073, 1.013 and 0.420, 0.203, 0.458), LC50(9.158, 9.218, 8.736 and 5.705, 6.482, 2.798) and LC90(467.65, 546.86, 523.49 and 809.5, 467.26, 86.99) values were observed for Et and Pp-AgNPs against all the adult flies, respectively. Et and Pp-AgNPs showed 75.0 and 85.0, 100.0 and 90.0 and 80.0 and 85.0 per cent of larval mortality for M. domestica, S. calcitrans and H. irritans irritans at 20 mg concentration after 48 hours exposure, respectively. The LC25(5.053, 0.0873, 1.5280 and 1.546, 1.166, 2.683), LC50(12.873, 2.294, 5.239 and 6.222, 3.508, 5.702) and LC90(76.086, 14.375, 54.485 and 87.715, 28.455, 23.871) values were indicated for all the three muscid larvae, respectively. The pupicidal assay indicated the highest per cent inhibition against S. calcitrans (45.0 and 40.0%) followed by M. domestica (30.0 and 30.0%) and H. irritans irritans (30.0 and 30.0%) at 8 mg concentration after 24 hours exposure for Et and Pp-AgNPs, respectively. The LC25(1.714, 0.412, 4.20 and 1.908, 3.00 and 4.757), LC50(6.270, 1.921, 30.386 and 28.229, 19.955 and 17.599) and LC90(73.675, 35.801, 62.32 and 4.719, 730.52 and 211.367) values were observed for Et and Pp-AgNPs against all the three pupae, respectively. Probit analysis revealed the significant effect with different concentration of AgNPs on adult, larvae and pupal mortality of muscid flies. The study indicated Et-AgNPs and Pp-AgNPs
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    ThesisItemOpen Access
    MOLECULAR CHARACTERISATION AND IDENTIFICATION OF IMMUNOREACTIVE POLYPEPTIDES OF FILARID NEMATODES OF DOGS
    (KARNATAKA VETERINARY, ANIMAL AND FISHERIES SCIENCIES UNIVERSITY, BIDAR, 2019) ARAVINDA, K.; ANANDA, K. J.
    A study on the molecular characterization and identification of immunoreactive polypeptides of filarid nematodes of dogs in Mangalore, Karnataka state was conducted. Different methods were used for detection of microfilariosis in dogs. A total of 250 dogs screened during the present study, 62 % harboured microfilaria. In the present study, three methods used for detection of microfilariosis in dogs. Modified knott’s method detected 94.19 % of the positive samples, direct wet smear and giemsa’s stained blood smear examination detected 61.93 and 57.42 % positive samples. Morphological studies of microfilaria were done by using giemsa’s and methylene blue phosphate staining techniques revealed microfilariae were unsheathed with blunt head and a tapering tail and cuticle appeared striated. The length and width of microfilaria when measured by giemsa’s and modified knott’s technique were 292.2 ± 1.267 μm, 334.9 ± 4.573 μm, 10.65 ± 0.068 μm and 6.862 ± 0.133 μm respectively. The histochemical staining technique revealed acid phosphatase activity at anal pore region of microfilaria by showing red azo dye indicating the microfilaria were of D. repens. Confirmatory of identification of species was done by PCR using species specific primers showing amplification of mitochondrial cytochrome oxidase subunit 1 (COI) gene with single amplicon 350 bp indicated D. repens. The protein profile of microfilarial and adult worm antigen of D. repens was evaluated using SDS-PAGE. The protein profile of microfilarial antigen showed six polypeptides ranging between 183 kDa to 29 kDa. A total of thirteen polypeptides were identified in adult worm antigen ranging from 208 kDa to 29 kDa. The immunoreactive polypeptides detected on western blot with microfilarial and adult worm antigen of D. repens using known positive serum with anti-dog IgG conjugate at 1:1000 dilution revealed 50 kDa and 29 kDa. Key words: Molecular characterization, Immunoreactive polypeptides, filarid nematodes, dog, Mangalore.
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    ThesisItemOpen Access
    EVALUATION OF LARVAL EXTRACTS OF CHRYSOMYA MAGGOTS FOR ANTIBACTERIAL PROPERTY
    (KARNATAKA VETERINARY, ANIMAL AND FISHERIES SCIENCES UNIVERSITY, BIDAR, 2018-08) GEETHA, N.; Dr. PLACID E DʼSOUZA
    In the present study, the larval extracts of Chrysomya maggots was tested for antibacterial property. The maggots were collected from wounds and carcasses in and around Bengaluru, India. The maggots from wounds were morphologically identified as C. bezziana and of carcasses were C. megacephala and C. albiceps. For larval extracts, the larvae were collected from the fly colony of C. megacephala which was successfully maintained in the laboratory, whereas larvae of C. bezziana collected from the fresh wounds due to their obligation of oviposition on fresh wounds. About two hundred, third instar larvae of Chrysomya specie
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    ThesisItemOpen Access
    SYSTEMATIC STUDY ON ARTHROPOD INFESTATION IN SHEEP WITH SPECIAL REFERENCE TO TICKS
    (KARNATAKA VETERINARY, ANIMAL AND FISHERIES SCIENCES UNIVERSITY, BIDAR, 2018-08) SUDHA RANI, R.; (Dr. PLACID E. D’SOUZA
    A systematic study on arthropod infestation in sheep in Karnataka based on season, breed, management system, agroclimatic zone and site preference was undertaken in nine organized and ten unorganised farms. The prevalence of ectoparasite infestations was 20.7% and 43.7% in organised and unorganized farms. Among all the ectoparasites, prevalence of ticks was highest followed by lice, mites, fleas, nasal bots and flies. The ticks were found resistant to deltamethrin, cypermethrin and were susceptible to amitraz using LPT and AIT-DD test. Among the two EPNs assessed, H.indica was found to be more virulent than S.abbasi. Among the phytoacaricides, Carica papaya seed extract was found to be more virulent than Ricinus communis leaves against ticks by LPT and AIT. By microscopy the prevalence rate of Theileria spp was 16.4% and 64% in organized and unorganized farms respectively. By PCR 24.7 % were positive for T.luwenshuni and 4.7% for T.ovis in organised farms. Also 72 % were positive for T.luwenshuni, 19 % positive for T.ovis and 4 % positive for B.ovis in unorganised farms. Staining of tick tissues with Geimsa revealed 13.5% and 10% of parasitic infection in organized and unorganized farms respectively. Similarly, MGP stain revealed 11.86% and 15.55% infection in organized and unorganized farms. Using PCR, the prevalence of T.luwenshuni in H.Kutchensis was 36% and of T.ovis in H.a.anatolicum was 4% in organised farms. Prevalence of T.luwenshuni and T.ovis was 77% in H.kutchensis and 47 % in H. a. anatolicum in unorganised farms. H.kutchensis ticks was considered as potential vector in disease transmission of T.luwenshuni and T.ovis in sheep in Karnataka. Key words: Arthropods prevalence, sheep farms, acaricides, phytoacaricides, EPNs, vector potentiality