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Open Access
Studies on synthetic pyrethroid resistance in Rhipicephalus (Boophilus) microplus and comparative sequence analysis of carboxylesterase gene
(G.B. Pant University of Agriculture and Technology, Pantnagar - 263145 (Uttarakhand), 2013-07) Arun, A.; Vatsya, Stuti
Two FAO recommended in vitro bioassays namely Adult Immersion Test (AIT) and Larval Packet Test (LPT) were employed to detect deltamethrin resistance in the tropical cattle tick, Rhipicephalus (Boophilus) microplus collected from tarai and hill regions of Uttarakhand. The AIT revealed the lowest LC50 (0.00085% A. I.; RF= 1) for Reference Susceptible R. microplus tick population. The highest LC50 (0.03842%; RF= 45.17) was observed for Dehradun (Mussourie) population of ticks followed by Nainital I (0.03403%; RF= 40.1), Nainital II (0.00571%; RF= 6.70) and Uttarkashi (0.00114%; RF= 1.29) ticks. In ticks collected from tarai region, the highest LC50 was found for Pantnagar II (0.03361%; RF= 39.53) population followed by Sitarganj (0.00663%; RF= 7.7), Pantnagar I (0.00652%; RF= 7.64), Ramnagar (0.00335%; RF= 3.88) and Tanakpur (0.00252%; RF= 2.94). The LC99 of different populations ranged from 0.3262% to 0.0036%. The R2 values determined from AIT ranged from 0.998 (Susceptible) to 0.847 (Nainital I ticks). The slopes of egg mass of different tick populations were highly negative. The values ranged from -146.95±20.96 (Susceptible) to -40.19±6.86 (Dehradun). The slopes of reproductive indices of different populations of ticks were found to be negative indicating the reduction of fecundity with increased concentration of acaricide. The values of slopes of inhibition of oviposition were positive and the highest value (34.7) was exhibited by Susceptible and lowest (6.92) by Dehradun ticks. In LPT bioassay, an LC50 of 0.00076% A. I. was observed for the Susceptible R. microplus population. Fom hills, Dehradun population exhibited the highest LC50 (0.03454%: RF= 45.39) followed by Nainital I (0.03113%: RF= 40.9), Nainital II (0.00421%; RF= 5.5) and Uttarkashi (0.00084%; RF= 1.1). From tarai region, the highest LC50 concentration was observed for Pantnagar II (0.03091%: RF= 40.65) population of ticks followed by Pantnagar I, Sitarganj, Ramnagar, and Tanakpur population of ticks (0.00493%; RF= 6.44), (0.00460%; RF= 6.05), (0.00250%; RF= 3.28) and (0.00164%; RF=2.10), respectively. The highest LC99 was found for Nainital I (1.8060%) population of ticks and the lowest for Susceptible population (0.0033%). Variations in LC50 values of different populations in AIT and LPT bioassays might be due to the difference in the stage of the tick being tested. The Chi square values ranged from 97.84 (Dehradun) to 18.298 (Nainital II). Six tick populations (Dehradun, Nainital I, Pantnagar II, Pantnagar I, Sitarganj and Nainital II) were found deltamethrin resistant (RF > 5), one tick population (Ramnagar) deltamethrin tolerant (RF 3-5) and two tick populations (Uttarkashi and Tanakpur) deltamethrin sensitive (RF< 3) in both in vitro bioassays. This implies that frequent monitoring of deltamethrin resistance in ticks is required so that timely measures could be taken before resistance to this acaricide becomes established. The Spearman Rank correlation coefficient (rS) was found to be 0.997 between AIT and LPT bioassays. The coefficient of determination (R2) was found to be 0.998. AIT and LPT were found highly correlated. The LPT bioassay takes 6 weeks to give results of acaricide resistance whereas AIT 2 weeks. So, it is suggested that AIT can be used as an effective screening test to detect the acaricide resistance in field populations of ticks. The sequences of amplified partial carboxylesterase gene (372 bp) of R. microplus from tick populations matched 100% with the carboxylesterase gene sequence of R. microplus available in GenBank. The sequence analysis revealed that no population had a nucleotide polymorphism at the position 300. The sequencing of partial carboxylesterase gene revealed four nucleotide changes in Uttarkashi and one in Dehradun population of ticks. The mutations in the nucleotide sequence of Uttarkashi population resulted in two amino acid mutations (Val 66 Gly, Asp 120 Asn) in the translated amino acid sequence. The amino acid sequence deduced from the nucleotide sequence of Dehradun population exhibited one (Glu 123 Arg) mutation. The mutations in Uttarkashi population had no effect in the resistance development as this population was phenotypically susceptible. Further investigations are required to identify role of mutation in resistance development in Dehradun population of ticks. Animal owners use different acaricides and adopt various application methods to control R. microplus. Thus frequent monitoring of acaricide resistance is very important to know the distribution and level of resistance to these acaricides. The baseline information thus generated will enable timely management of spread of acaricide resistance in ticks and hence enhance animal productivity.
Open Access
Evaluation of protective effect of Erythrina variegata and Spirulina platensis in imidacloprid intoxicated WLH cockerels
(G.B. Pant University of Agriculture and Technology, Pantnagar - 263145 (Uttarakhand), 2014-07) Gupta, Mayank; Singh, S.P.
This study was undertaken to investigate the ameliorative efficacy of Erythrina variegata leaf powder (EVLP) @1% and Spirulina platensis (SP) @ 0.2% in feed by determining hematobiochemical and antioxidant parameters and histopathological examination following simultaneous administration of imidacloprid (IMI) @ 50 ppm in feed for 60 days in white leghorn cockerels. Hydroethanolic extract of leaves of Erythrina variegata (HEEV) was prepared for phytochemical analysis, and to evaluate its effect on isolated smooth muscles of rat ileum. Phytochemical analysis revealed the presence of alkaloids, flavonoids, proteins, sterols, saponins and terpenes. For evaluation of protective effects of SP and EVLP in IMI intoxicated cockerels, forty two male white leghorn chicks were divided equally and randomly into six groups viz. I, II, III, IV, V and VI. Group I served as control and other groups were administered as SP @ 0.2% in II, IMI @ 50 PPM in III, SP @ 0.2% plus IMI @ 50 PPM in group IV, EVLP (1%) in group V and EVLP (1%) plus IMI @ 50 PPM in VI, respectively, for 60 days and parameters were recorded at 0, 30 and 60 days interval. A significant (P<0.05) decline in body weight gain was measured in imidacloprid treated cockerels, whereas, SP and EVLP treatments alone in groups II and V and simultaneously with IMI in groups IV and VI revealed an improvement in body weight gain. A significant (P<0.05) reduction in Hb, PCV, TEC, TLC was observedin group III and a significant (P<0.05) increase in Hb, PCV, TEC, TLC was observed in group II as compared to control after 60 days. Groups IV and VI showed a significant (P<0.05) improvement in Hb, PCV, TEC, TLC as compared to group III. A significant decline in total serum protein, albumin and globulin was reported in imidacloprid treated cockerels of group III as compared to control. Groups IV and VI showed significant (P<0.05) increase in total serum protein, albumin and globulin as compared to group III showing ameliorative effect of SP and EVLP medication. A significant (P<0.05) increase in triglycerides, cholesterol, total bilirubin, indirect bilirubin, creatinine, AST, ALT, ALP was observed in cockerels of imidacloprid treated Group III, whereas, a significant (P<0.05) decline in value of these parameters was observed in groups IV and VI supplemented with SP and EVLP, respectively. A significant (P<0.05) decline in RBC catalase and SOD and an increase in LPO in RBC, liver, kidney and testes was observed in group III which, however, returned to normalcy following simultaneous medication with SP and EVLP in groups IV and VI after 60 days. Histopathological changes such as severe congestion of blood vessels, interstitial hemorrhages between tubules, swelling of the glomeruli and hypercellularity of the glomerular capillaries in kidney; severe congestion of large blood vessel, sinusoidal congestion, vacuolar degeneration of the hepatocytes throughout the parenchyma and presence of mononuclear cells around congested blood vessels in liver; reduction in size of the seminiferous tubules with a single layer of germinal epithelium in testes and congestion of blood vessels, elongation of neurons, gliosis and satellitosis in brain were observed in IMI treated group after 60 days trial in cockerels.The tissues from IMI cockerels supplemented with EVLP in group VI showed mild pathological lesions while there were no pathological changes in liver, kidney, testes and brain from IMI plus SP supplemented cockerels of group IV. Presensitization with HEEV (100-1200 µg/ml) for five minutes showed significant spasmolytic activity against acetylcholine (10-5M) and carbachol (10-5M) induced contractions in isolated rat ileum It is concluded from this study that administration of imidacloprid (50 ppm) in feed for 60 days produced haemotoxic, hepatotoxic, nephrotoxic and potent oxidative effects, which were ameliorated following simultaneous administration of Spirulina platensis (0.2%) and Erythrina variegata (1%), respectively, for 60 days in white leghorn cockerels. Hydroethanolic extract of Erythrina variegata showed spasmolytic activity in rat ileum.
Open Access
Comparative efficacy of madar (Calotropis procera) and Amprolium on coccidiosis in commercial broilers
(G.B. Pant University of Agriculture and Technology, Pantnagar - 263145 (Uttarakhand), 2015-05) Chauhan, Sakshi; Singh, Vidya Sagar
Coccidiosis is one of the major harmful disease of universal importance in poultry. Present investigation was conducted to evaluate the comparative anticoccidial efficacy of madar leaf powder and amprolium supplementation against mixed Eimeria sp. infection. For the purpose, 168, day old broiler chicks were divided into 7 groups, with 2 replicates of 12 chicks each. Broilers of group I and II were provided unsupplemented diet. Broilers of group III and IV were provided 0.0125% amprolium supplemented diet, whereas broilers of group V and VI were provided diet supplemented with 0.2% madar leaf powder and broilers of group VII were provided 0.4% madar leaf powder supplemented diet. On 15th day of experiment, broilers of group II, IV, VI and VII were infected with 50,000 mixed Eimeria sp. sporulated oocysts. Experimenrt was conducted for 30 days period. Growth performance for pre-infection period indicated significant effect of 0.4% madar leaf powder on growth performance followed by 0.2% madar leaf powder. During post infection period coccidiosis caused significant reduction in growth performance in infected groups. Growth performance restoration was maximum in amprolium supplemented group, followed by 0.4% madar leaf powder and 0.2% madar leaf powder supplemented groups. Overall growth performance was non significantly different between infected amprolium supplemented and 0.4% madar leaf powder supplemented group, however 0.2% madar leaf powder supplemented group showed significant difference to amprolium supplemented group. Haematological parameters on 0 DPI and 15 DPI were non significant among all groups. On 5 and 10 DPI, Hb, PCV, TEC, MCH, MCHC, heterophils % and monocytes % were decreased, whereas MCV, TLC, lymphocytes % and eosinophils % were increased in infected groups than respective uninfected groups. Haematological variation due to coccidiosis was maximum in control infected group. Maximum restoration of normal parameters was observed in amprolium supplemented group followed by 0.4% and 0.2% madar leaf powder supplemented group. Parasitological parameters in terms of percent faecal score, percent survival, percent weight gain, performance index, average oocyst production, percent reduction in oocyst production, average oocyst index, average lesion score, percent protection against lesion, global index and percent global index clearly indicated maximum anticoccidial effect of amprolium, however anticoccidial effect of 0.4% as well as 0.2% madar leaf powder was also comparable to amprolium. All supplementations showed very good efficacy against coccidiosis as indicated by percent global index. Carcass traits as dressed yield % and organ weight % were significantly higher in 0.4% as well as 0.2% madar leaf powder supplemented groups followed by amprolium supplemented and non supplemented groups. Difference in the dressed yield % among respective infected and uninfected groups was restored on 10 and 15 DPI, but organ weights % were significantly higher in broilers of respective infected groups than uninfected groups on 5, 10 and 15 DPI. Histopathological changes revealed maximum degenerative changes and presence of maximum parasitic stages as second generation schizonts, macrogametocyte and oocysts on 5, 10 and 15 DPI in caeca of control infected group whereas minimum deviation in caecal architecture was noticed in amprolium supplemented group and parasitic stages were absent on 10 and 15 DPI. 0.4% as well as 0.2% madar leaf powder supplemented group also had significantly lower architectural changes than control infected group and parasitic stages were absent on 15 DPI. From the results of present study it can be concluded that madar (Calotropis procera) leaf powder supplementation had anticoccidial efficacy comparable to amprolium supplementation and further research is needed to evaluate the coccidiosis curative property of madar leaf powder.
Open Access
Studies on evaluation of some phytoacaricides against Rhipicephalus (Boophilus) microplus
(G.B. Pant University of Agriculture and Technology, Pantnagar - 263145 (Uttarakhand), 2017-08) Vijay Kumar; Vatsya, Stuti
A study was under taken with objective of detecting synthetic pyrethroid resistance (deltamethrin concentrations- 0.0003125%, 0.000625%, 0.00125%, 0.0025%, 0.005%, 0.01% and 0.02%-each tested in triplicate) in R. microplus using two in vitro laboratory bioassays namely Adult Immersion Test (AIT) and Larval Packet Test (LPT) collected from different parts of Uttarakhand state {Nainital (Bindu Khatta, Patwadangar, Halduchor), Udham Singh Nagar (Saanp Katani Khatta, Pantnagar Dairy, Nehru Colony, Sitarganj), Pithoragarh (Bin) and Uttarkashi} to generate a baseline data and Azadirachta and Eucalyptus plants were screened for acaricidal activity. The susceptible tick population collected from Bindu Khatta, Nainital displayed an LC50 of 0.00075% Active Ingredient (A.I.) [Confidence Interval (C.I.) 95% 0.0006-0.0009] and LC99 of 0.00533% (C.I. 95% 0.0034-0.0112) in AIT bioassay. The R. microplus population from Nehru colony exhibited the highest LC50 of 0.03192%, (CI 95% 0.0188-0.1026%) followed by Pithoragarh (0.00526%, CI 95% 0.0036-0.0083%), Pantnagar dairy (0.00514%, CI 95% 0.0036-0.0080%), Sitarganj (0.00376%, CI 95% 0.0025-0.0061%), Halduchor (0.00337% CI 95% 0.0025-0.0047%), Saanp Katani Khatta (0.00189% CI 95% 0.0013-0.0026%), Nainital (Patwadangar) (0.00177% CI 95% 0.0012-0.0025%) and Uttarkashi (0.00088%, CI 95% 0.0007-0.0011). The R2 values of 0.994, 0.991, 0.978, 0.974, 0.973, 0.964, 0.962, 0.952 and 0.877 were detected in Bindu Khatta, Uttarkashi, Pithoragarh, Pantnagar Dairy, Nehru colony, Nainital, Halduchor, Sitarganj, and Saanp Katani Khatta populations, respectively. The Nehru colony population of ticks had the highest resistance factor (RF) of 42.55, whereas the Pithoragarh and Pantnagar dairy populations had RF values of 7.01 and 6.85, respectively; Sitarganj, Halduchor, Saanp Katani Khatta and Nainital tick populations were 5.01, 4.49, 2.52, and 2.36 times resistant, respectively. Level IV resistance was found in Nehru colony tick population; level II in Pithoragarh, Pantnagar Dairy, Sitarganj and level I in Halduchor, Saanp Katani Khatta and Nainital R. microplus populations. The population of ticks collected from Uttarkashi and Bindu Khatta were found susceptible to deltamethrin. In LPT bioassay, an LC50 of 0.00074% A. I. (C.I. 95% 0.0005-0.0010) was observed for the Susceptible R. microplus population. The LC50 values were observed to be (0.03524%, CI 95% 0.0248- 0.0607%), (0.00508%, CI 95% 0.0037-0.0073%), (0.00439%, CI 95% 0.0030-0.0067), (0.00393%, CI 95% 0.0021-0.0089%), (0.00365%, CI 95% 0.0025- 0.0054%), (0.00196%, CI 95% 0.0013-0.0029%), (0.00165% CI 95% 0.0012-0.0022%) and (0.00097% CI 95% 0.0006-0.0015) for Nehru colony, Pantnagar Dairy, Sitarganj, Pithoragarh, Halduchor, Saanp Katani Khatta, Nainital and Uttarkashi, respectively. Azadirachta indica and Eucalyptus citriodora were selected to know their acaricidal potential against R. microplus ticks. The % yield obtained with, aqueous, methanolic and aqueous-methanolic extract was, 16, 7.48 and 17.56 for A. indica and 13.28, 18.96 and 19.28 for Eucalyptus. The results of AIT using the aqueous, methanolic and aqueous-methanolic extract of both plants revealed a dose dependent decrease in reproductive index (RI) and percent inhibition of oviposition (IO%) generally from concentration 2.5 to 20%. A decrease in egg production was significant (p<0.05) for 2.5, 5, 10 and 20% concentration of the extract. Tick mortality percent of 40 and 70% was observed at 10 and 20 % concentration of methanolic extract solution of A. indica. The results of AIT with E. citriodora revealed that RI and IO% decreased from concentration 1.25 to 20%. The results of AIT using the Neem oil showed that there was dose dependent decrease in RI and IO% in concentrations ranging from 1.25 to 20%. A decrease of 5, 30, 40, 50 and 60% hatching of eggs was observed at 1.25, 2.5, 5, 10 and 20% concentration of oil, respectively. With Eucalyptus oil, also a dose dependent decrease in RI and IO% for 1.25% concentration was noted. A decrease of 50% hatching of eggs was also observed at 1.25% concentration of oil. Ticks failed to oviposit at 2.5% to 20% concentration. 100% mortality was observed at 2.5% to 20% concentration of oil. In LPT bioassay, an LC50 of 9.333% A. I. (C.I. 95% 6.132-16.983) was observed for the Azadirachta aqueous extract solution against resistant R. microplus population. With E. citriodora aqueous methanolic solution, LC50 of 9.620% A. I. (C.I. 95% 4.491-17.148), methanolic LC50 of 14.492% A. I. (C.I. 95% 13.210-16.060) and aqueous LC50 of 17.556% A. I. (C.I. 95% 2.916.210-18.817)}. E. citriodora oil exhibited LC50 of 11.634% and neem oil an LC50 of 22.818% against larvae of R. microplus population. In LIT, no mortality of R. microplus larvae was recorded at any concentration of A. indica and E. citriodora extracts except at 20% concentration of neem aqueous solution (85%). Several phytoconstituents were revealed on phytochemical analysis of A. indica [alkaloids, glycosides, saponins, sterols, tannins and triterpenes] and E. citriodora [alkaloids, flavonoids, reducing sugar, saponins, tannins and triterpenes]. Based on results of laboratory bioassays, it is possible to conclude that the oils of A. indica and E. citriodora caused a negative effect on reproduction and hatchability against deltamethrin resistant R. microplus while Eucalyptus oil even resulted in mortality of ticks. The use of these oils shows great potential for the future as an alternative biocontrol method for R. microplus. These can be incorporated as part of an integrated control programme for ticks.
Open Access
Prevalence of gastro-intestinal parasites in zoo animals of some Indian zoos
(G.B. Pant University of Agriculture and Technology, Pantnagar - 263145 (Uttarakhand), 2017-07) Niranjan, Anuruddha Singh; Singh, Vidya Sagar
The present research was conducted to study the prevalence and intensity of gastrointestinal parasites in the Zoo animals at Zoological Park, Nainital and Kanpur. A total of 333 faecal samples (99 samples from Nainital, Zoo and 234 samples from Kanpur, Zoo) were randomly collected. The overall prevalence of parasitic infection were 28.28% and 32.05% whereas, prevalence of helminths, protozoans and mixed infection were 20.20%, 7.07%, 1.01% and 20.94%, 8.11%, 2.99% for Zoological Parks, Nainital and Kanpur respectively. The results for Zoological Park, Nainital indicated that the helminth infections were more compared to protozoans and mixed infection in herbivores, carnivores and some birds. The prevalence of helminths and protozoans in carnivores were 20.83%, 4.16% respectively and for herbivores were 27.77%, 16.66% and mixed infection 5.55% respectively. Gastrointestinal parasites (eggs/cysts/oocysts) identified in herbivores were Strongyle sp., Capillaria sp. (11.11%), Trichuris sp., Muellerius sp. (5.55%) and Eimeria sp. (16.66%). EPG/CPG/OPG ranged from 100 to 500. Gastrointestinal parasites (eggs/cysts/oocysts) identified in carnivores were Toxocara cati (12.50%), Toxocara canis (8.33%) and Isospora sp.(4.16%). EPG/CPG/OPG ranged from 150 to 450. Common Peafowls were found positive with Ascaridia galli (18.18%) and Eimeria sp. (3.63%). The results for Zoological Park, Kanpur indicated that the helminth infections were more than protozoan and mixed infection in herbivores, carnivores, omnivores and non-human primates. The prevalence of helminths, protozoans and mixed infection in carnivores were 34.42%, 6.55% and 3.27% respectively, for herbivores, 16.93%, 4.83% and 3.22% repectively, for non-human primates 13.33% and 20%. Gastrointestinal parasites (eggs/cysts/oocysts) identified in herbivores were Amphistome (2.41%), Fasciola sp. (1.61%), Trichuris sp. (3.22%), Strongyle sp., Strongyloides sp., Oesophagostomum sp., Trichostrongylus sp.(2.41%) and Eimeria sp. (4.83%). EPG/CPG/OPG was ranged from 50to350. Gastrointestinal parasites (eggs/cysts/oocysts) identified in carnivores were Ancylostoma sp. (8.19%), Toxocara cati (8.19%), Strongyloides sp. (6.55%), Trichuris sp. (4.91%) Toxocara canis (3.27%), Toxascaris leonina (1.63%), Spirometra sp. (1.63%) and Isospora sp. (6.55%). EPG/CPG/OPG were ranged from 100 to 600. Gastrointestinal parasites (eggs/cysts/oocysts) identified in non-human primates were Strongyloides sp. (13.33%) and Eimeria sp. (20%). EPG/CPG/OPG were ranged from 200 to 350. The overall prevalence of intestinal parasitic infection in the present study was 30.93% with 21.02% samples positive with helminths, 7.80% samples positive with protozoans and 2.10% samples positive with mixed infection. The overall prevalence of helminths, protozoans and mixed infection in herbivore were 18.30%, 6.33% and 3.52%, in carnivores were 30.58%, 5.88% and 2.35%, in non-human primates helminths and protozoan prevalence were 12.76% and 21.27% and in wild birds were 18.18% and 3.63% respectively. The overall prevalence of helminths in omnivores was 0.60%. There was no protozoan and mixed infection. Looking to the incidence of gastro-intestinal parasites kept in captivity it can be concluded that there is a need for detailed epidemiological investigation on the prevalence of gastrointestinal parasites by regular screening of faecal samples in Zoo animals with respect to season, age, climate etc. Based on the prevalence of parasites deworming with suitable anthelmintic is required. Proper hygienic measures for the control of different stages of parasites need to be taken care of so as to reduce the parasitic infection in Zoo animals.
Open Access
Studies on evaluation of some phytoacaricides against Rhipicephalus (Boophilus) microplus
(G.B. Pant University of Agriculture and Technology, Pantnagar - 263145 (Uttarakhand), 2017-08) Vijay Kumar; Vatsya, Stuti
A study was under taken with objective of detecting synthetic pyrethroid resistance (deltamethrin concentrations- 0.0003125%, 0.000625%, 0.00125%, 0.0025%, 0.005%, 0.01% and 0.02%-each tested in triplicate) in R. microplus using two in vitro laboratory bioassays namely Adult Immersion Test (AIT) and Larval Packet Test (LPT) collected from different parts of Uttarakhand state {Nainital (Bindu Khatta, Patwadangar, Halduchor), Udham Singh Nagar (Saanp Katani Khatta, Pantnagar Dairy, Nehru Colony, Sitarganj), Pithoragarh (Bin) and Uttarkashi} to generate a baseline data and Azadirachta and Eucalyptus plants were screened for acaricidal activity. The susceptible tick population collected from Bindu Khatta, Nainital displayed an LC50 of 0.00075% Active Ingredient (A.I.) [Confidence Interval (C.I.) 95% 0.0006-0.0009] and LC99 of 0.00533% (C.I. 95% 0.0034-0.0112) in AIT bioassay. The R. microplus population from Nehru colony exhibited the highest LC50 of 0.03192%, (CI 95% 0.0188-0.1026%) followed by Pithoragarh (0.00526%, CI 95% 0.0036-0.0083%), Pantnagar dairy (0.00514%, CI 95% 0.0036-0.0080%), Sitarganj (0.00376%, CI 95% 0.0025-0.0061%), Halduchor (0.00337% CI 95% 0.0025-0.0047%), Saanp Katani Khatta (0.00189% CI 95% 0.0013-0.0026%), Nainital (Patwadangar) (0.00177% CI 95% 0.0012-0.0025%) and Uttarkashi (0.00088%, CI 95% 0.0007-0.0011). The R2 values of 0.994, 0.991, 0.978, 0.974, 0.973, 0.964, 0.962, 0.952 and 0.877 were detected in Bindu Khatta, Uttarkashi, Pithoragarh, Pantnagar Dairy, Nehru colony, Nainital, Halduchor, Sitarganj, and Saanp Katani Khatta populations, respectively. The Nehru colony population of ticks had the highest resistance factor (RF) of 42.55, whereas the Pithoragarh and Pantnagar dairy populations had RF values of 7.01 and 6.85, respectively; Sitarganj, Halduchor, Saanp Katani Khatta and Nainital tick populations were 5.01, 4.49, 2.52, and 2.36 times resistant, respectively. Level IV resistance was found in Nehru colony tick population; level II in Pithoragarh, Pantnagar Dairy, Sitarganj and level I in Halduchor, Saanp Katani Khatta and Nainital R. microplus populations. The population of ticks collected from Uttarkashi and Bindu Khatta were found susceptible to deltamethrin. In LPT bioassay, an LC50 of 0.00074% A. I. (C.I. 95% 0.0005-0.0010) was observed for the Susceptible R. microplus population. The LC50 values were observed to be (0.03524%, CI 95% 0.0248- 0.0607%), (0.00508%, CI 95% 0.0037-0.0073%), (0.00439%, CI 95% 0.0030-0.0067), (0.00393%, CI 95% 0.0021-0.0089%), (0.00365%, CI 95% 0.0025- 0.0054%), (0.00196%, CI 95% 0.0013-0.0029%), (0.00165% CI 95% 0.0012-0.0022%) and (0.00097% CI 95% 0.0006-0.0015) for Nehru colony, Pantnagar Dairy, Sitarganj, Pithoragarh, Halduchor, Saanp Katani Khatta, Nainital and Uttarkashi, respectively. Azadirachta indica and Eucalyptus citriodora were selected to know their acaricidal potential against R. microplus ticks. The % yield obtained with, aqueous, methanolic and aqueous-methanolic extract was, 16, 7.48 and 17.56 for A. indica and 13.28, 18.96 and 19.28 for Eucalyptus. The results of AIT using the aqueous, methanolic and aqueous-methanolic extract of both plants revealed a dose dependent decrease in reproductive index (RI) and percent inhibition of oviposition (IO%) generally from concentration 2.5 to 20%. A decrease in egg production was significant (p<0.05) for 2.5, 5, 10 and 20% concentration of the extract. Tick mortality percent of 40 and 70% was observed at 10 and 20 % concentration of methanolic extract solution of A. indica. The results of AIT with E. citriodora revealed that RI and IO% decreased from concentration 1.25 to 20%. The results of AIT using the Neem oil showed that there was dose dependent decrease in RI and IO% in concentrations ranging from 1.25 to 20%. A decrease of 5, 30, 40, 50 and 60% hatching of eggs was observed at 1.25, 2.5, 5, 10 and 20% concentration of oil, respectively. With Eucalyptus oil, also a dose dependent decrease in RI and IO% for 1.25% concentration was noted. A decrease of 50% hatching of eggs was also observed at 1.25% concentration of oil. Ticks failed to oviposit at 2.5% to 20% concentration. 100% mortality was observed at 2.5% to 20% concentration of oil. In LPT bioassay, an LC50 of 9.333% A. I. (C.I. 95% 6.132-16.983) was observed for the Azadirachta aqueous extract solution against resistant R. microplus population. With E. citriodora aqueous methanolic solution, LC50 of 9.620% A. I. (C.I. 95% 4.491-17.148), methanolic LC50 of 14.492% A. I. (C.I. 95% 13.210-16.060) and aqueous LC50 of 17.556% A. I. (C.I. 95% 2.916.210-18.817)}. E. citriodora oil exhibited LC50 of 11.634% and neem oil an LC50 of 22.818% against larvae of R. microplus population. In LIT, no mortality of R. microplus larvae was recorded at any concentration of A. indica and E. citriodora extracts except at 20% concentration of neem aqueous solution (85%). Several phytoconstituents were revealed on phytochemical analysis of A. indica [alkaloids, glycosides, saponins, sterols, tannins and triterpenes] and E. citriodora [alkaloids, flavonoids, reducing sugar, saponins, tannins and triterpenes]. Based on results of laboratory bioassays, it is possible to conclude that the oils of A. indica and E. citriodora caused a negative effect on reproduction and hatchability against deltamethrin resistant R. microplus while Eucalyptus oil even resulted in mortality of ticks. The use of these oils shows great potential for the future as an alternative biocontrol method for R. microplus. These can be incorporated as part of an integrated control programme for ticks.
Open Access
Studies on evaluation of efficacy of herbal plants against gastrointestinal nematodosis in goats
(G.B. Pant University of Agriculture and Technology, Pantnagar - 263145 (Uttarakhand), 2016-06) Sastya, Sushmita; Rajeev Ranjan Kumar
Open Access
Epidemiology and anthelmintic resistance of gastrointestinal nematodes in small ruminants with special reference to evaluation and characterization of anthelmintic activity of some herbal plants
(G.B. Pant University of Agriculture and Technology, Pantnagar - 263145 (Uttarakhand), 2014-08) Rajeev Ranjan Kumar; Yadav, C.L.
The present study was planned to know the distribution of G.I. nematodes, evaluate the efficacy of commonly used anthelmintics against G.I. nematodes of small ruminants and also exploit some herbal plants as an alternative treatment. The epidemiology of G.I. nematodes of small ruminants was studied in district Udham Singh Nagar of Uttarakhand state. Small ruminants were found to be infected with G.I. nematodes throughout the year with slightly higher overall prevalence rate (84.85%) in goats as compared to sheep (81.25%). Higher prevalence of strongyle worms was observed in rainy season (99.81% and 98.60%) and lowest (33.01% and 77.80%) during the month of winter season in both sheep and goat. Higher mean epg of strongyle worms was recorded in goats (605.18) as compared to sheep (579.50) Coproculture studies revealed the presence of H. contortus, O. columbianum, T. colubriformis and S. papillosus throughout the year in both sheep and goats. In the present study, H. contortus was predominat G.I. strongyle worm of small ruminants. The highest (47) pasture larval (L3) burden was observed in the month of August and lowest (7) in February. In sheep and goats, highest (379 and 243) post mortem worm burden was recorded in the month of September. Anthelmintic resistance was detected by FECRT and results showed that sheep of Shamaliti, Kedarkatha and goats of Gwaldum were found resistant to fenbendazole ( FECR%: -170, -10 and 63) and tetramisole (FECR%: 18, 24 and 91) while all sheep and goats were found susceptible to ivermectin with FECR% varied from 97-100%. Post coproculture examination in all animals revealed the presence of 100% H. contortus as resistant nematode. In-vitro trial of herbal plants showed that average highest corrected mortality of 95%, 100%, 100% and 100% of A. indica was observed at 0.25%, 0.5%, 1% and 2%, respectively. However, lowest percent corrected mortality of 0, 0, 20 and 60 of P. persica was recorded at 0.25%, 0.5%, 1% and 2%, respectively. Overall A. indica possed highest anthelmintic activity (corrected mortality-97.0%) followed by T. Copticum (corrected mortality-95%) against H. contortus. Phytochemical analysis revealed that tannins, saponins, alkaloids, flavonoids, reducing sugars, resins, sterols, glycosides and triterpenes were the common constituents found in various extracts of different tested plants. In-vitro trial of various plants indicated that A. indica had highest anthelmintic activity. So that, phytochemicals found in the plant were further fractionated by HPTLC. Phytochemical analysis of A. Indica showed that alkaloids and tannins were found in all extracts (aqueous, diethyl ether and methanol) whereas saponins, triterpenes and reducing sugars were found in diethyl ether and methanol extracts. However, glycosides showed its presence only in methanol extract. Out of all three extracts of A. indica, methanol extract showed maximum in-vitro efficacy against H. contortus followed by aqueous and least by diethyl ether extract. Alkaloids of methanol, aqueous and diethyl ether extracts were fractionated into 7, 3 and 7 peaks, respectively upon HPTLC. The tannins of above extracts were fractionated into 3, 1 and 7 peaks, respectively. On the basis of present study, it is suggests that epidemiological data may be used to formulate effective control strategies against G.I. nematodosis in small ruminants. And avoid the use of resistant drugs for controlling G.I. nematodosis in small ruminants of organized farms in Uttarakhand. Leaves of A. Indica and seeds of T. copticum may be used as an alternative treatment against most pathogenic H. contortus parasite.
Open Access
Epidemiology of paramphistomosis in ruminants of Uttarakhand and comparative efficacy of some ethnomedicinal plants
(G.B. Pant University of Agriculture and Technology, Pantnagar - 263145 (Uttarakhand), 2014-01) Chaudhary, Sanjay; Gupta, H.P.
An epidemiological study of paramphistomosis of domestic ruminants (cattle buffalo, sheep and goat) was carried out in Garhwal region of Uttarakhand during 2009-11. A total of 6241 faecal samples were screened with an overall percent prevalence of 10.06 (13.33% cattle, 10.57 % buffalo, 8.40% sheep and 7.68% goats). An overall 1.88 mean eggs per gram of faeces (EPG) (cattle 2.68, buffalo 2.16, sheep 1.34 and goats 1.3) was recorded. Overall monthly highest prevalence of paramphistomosis was observed in July (16.12%) followed by June (15.48%) and August (15.7%) and lowest was recorded in December (4.6%), January (5.82%) and February (7.03%) with highest mean EPG being observed in August (3.65) and July (3.51) and lowest during December (0.7) and November (1.12). In cattle, highest monthly prevalence was found in August (25.33%) and lowest in December (5.3%). In buffaloes, the highest percentage of infection was observed in July (16.0%) and lowest in December (7.07%). In sheep, the highest monthly infection was recorded in July (16.36%) and lowest in November (1.33%). The same in goats were recorded in June (12.67 %) and December (3.33%), respectively. Maximum overall seasonal prevalence was observed in monsoon and post monsoon (12.91%) followed by summer (11.3%) and least in winter (6.06 %). The paramphistome species identified included Paramphistomum cervi, Fischoederius elongatus, Cotylophoron cotylophoroum and Gastrothylax crumeniform from rumen and reticulum and Gigantocotyle explanatum from bile ducts. Overall 18.57% (20.69% buffaloes, 19.68% sheep and 14.74% goat) of slaughtered ruminants were found to be positive for mature paramphistomes. The results of epidemiological study revealed that the domestic ruminants of Tehri - Garhwal region of Uttarakhand suffered from paramphistomosis throughout the year and certainly it cannot be treated as a casual entity. Therefore to control paramphistomosis in this area, it is suggested to treat animals with antitrematodal drug twice in a year- once in the beginning of summer (April/May) and second at the end of monsoon season (July/August/September). Six medicinal plants namely Emblia ribes, Butea frondosa, Andrographis paniculata, Argemone maxicana, Azardichta indica and Prunus persica were evaluated for their in vitro efficacy against P. cervi and analysed for presence of different phytochemical groups and fractionated through HPTLC. The maximum cumulative antitrematodal efficacy (80%) was observed by chloroform extracts of E. ribes and methanol extract of A. maxicana followed by aqueous and methanol extracts of B. frondosa (73.33%) at 2% concentration whereas at 3%, maximum efficacy (83.33%) was observed in aqueous and chloroform extracts of E. ribes followed by B. frondosa (76.67%). Therefore, E.ribes and B .frondosa may be explored their antitrematodal activity. Maximum histopathological changes (vacuole formation and necrosis etc.) in tagument and other internal organs, were also observed in the sections of paramphistome exposed to above mention plants extracts. Phytochemical analysis revealed tannins in all extracts except aqueous extract of A. maxicana. Coumarins was present in chloroform and methanol extracts of A. paniculata and E. ribes; chloroform and aqueous extracts of A. mexicana and all extracts of B. frondosa. Triterpenes were found in chloroform and methanol extracts. Sterols were found in only five extracts (aqueous extract of A. paniculata, methanol and aqueous extracts of A. Maxicana and chloroform and methanol extracts of B. frondosa). Glycosides were present in chloroform extract of A. paniculata and E. ribes, methanol extract of A. maxicana and aqueous extract of E. ribes; proteins in aqueous extract of A. paniculata and B. frondosa and methanol extract of B. frondosa and A. paniculata and saponins in aqueous extract of A. paniculata and B. frondosa and chloroform extract of E. ribes and B. frondosa. Reducing sugars were found chloroform extract of A. paniculata and A. maxicana and aqueous extract of E. ribes. Alkaloids were observed in chloroform, methanol and aqueous extracts of A. maxicana and A. paniculata and only in aqueous extract of E. ribes. Flavonoids were found only in methanol extract of A. maxicana. On densitometric analysis the above mentioned different phytochemicals groups were fractionated by HPTLC. Plants like A. paniculata, B. frondosa, A. maxicana, E. ribes had components like tannins and/or alkaloids which have anthelmintic properties and can be exploited as dewormers or as neutraceuticals.

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