Browsing by Author "Narang, Gulshan"
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ThesisItem Open Access Effect Of Infectious Bursal Disease Vaccine And Levamisole On Pathology Of Hydropericardium Syndrome In Broiler Chick(Chaudhary Charan Singh Haryana Agricultural University;Hisar, 2004) Narang, Gulshan; Pruthi, A.K.ThesisItem Open Access Effect of Levamisole on Development of HVT Vaccinal Immunity Against Markes Disease(College of Veterinary Sciences Chaudhary Charan Singh Haryana Agricultural University Hisar, 1991) Narang, Gulshan; Kharole, M. UThesisItem Open Access Effect of Spirulina feeding on sequential pathology of Infectious bursal disease vaccine in broiler chicken(LUVAS, 2018) Pravesh Kumari; Narang, GulshanInfectious bursal disease is an acute, highly contagious viral infection of chickens manifested by inflammation and subsequent atrophy of the bursa of Fabricius and various degrees of immunosuppression. A number of vaccination strategies have been applied in the field to control IBD. Live commercially available hot strains of vaccines for IBD lead to varying levels of immunosuppression which increases the bird’s vulnerability to various infections. The present study was conducted to observe the effect of Spirulina feeding on sequential pathology of Infectious bursal disease vaccine (IBDV) in broiler chickens. One hundred and two day old chickens were reared up to 38th day of age. At the age of 10 days, chickens were divided randomly into four groups (groups A (33), B (27), C (21) and D (21) having different number of chickens. From 10 to 20 days, feed of all the chickens of group B and D was supplemented with probiotic Spirulina at the dose rate of 1.0 % of feed (i.e. 1.0 g/ 100 gm of feed) whereas all chickens of group A and C were given feed without Spirulina supplementation. All the chickens of group C and D were vaccinated with IBDV intermediate plus strain vaccine at the age of 17 days whereas no vaccine was given to the chickens of groups A, and B. Mild clinical signs were observed in broiler chickens after vaccination in vaccinated groups. Chickens in group C (IBDV vaccinated) showed mild dullness, depression, anorexia with reduced feed and water intake. Whitish coloured diarrhoea was also observed in few chickens at 7 and 14 day post vaccination (DPV). However in group D chickens (vaccinated and Spirulina fed) above mentioned clinical signs were milder in nature and were also observed only in few chickens throughout the experiment. Mean body weight in Spirulina fed group B was found to be significantly higher among all the groups. Mean body weight of group C was found significantly lower among all the groups. Spirulina supplementation increased the mean body weight in group D. Mean values of bursal index was higher in group B as compared to control group. There was significant reduction in bursal index in IBD vaccination. Spirulina was able to significantly increase the bursal index in IBD vaccinated chickens. There was higher concentration of serum total protein and albumin in Spirulina fed group B. There was significant lower concentration of serum total protein and albumin in IBD vaccinate group chickens. Spirulina supplementation was able to slightly reduce the effect of IBD vaccine on total protein and albumin level. There was significantly lower activity of serum aspartate transaminase in Spirulina fed group however significantly higher activity was observed in IBD vaccinated alone group indicating that Spirulina supplementations in IBD vaccinated chickens was able to reduce the activity of this enzyme. IBD vaccination led to significantly higher activity of alanine transaminase in vaccinated chickens. Spirulina supplementation in IBDV vaccinated chickens helped to reduce the activity of this enzyme to some extent. A significant higher creatinine concentration was observed in IBDV vaccinated groups (C and D). Spirulina supplementation in IBD vaccinated chickens helped to keep the concentration of creatinine at lower levels. Mean blood urea nitrogen concentration (BUN) concentrations were slightly higher in the vaccinated groups (C and D). Amongst vaccinated groups (C and D) little higher values of serum BUN level was found in group C as compared to group D. Serum ALP activity was found to be significantly higher in group C (IBDV vaccinated). However Spirulina supplementation in IBD vaccinated chickens reduces the activity of this enzyme to some extent. SOD activity was found significantly lower in the vaccinated group chickens whereas Spirulina supplementation in IBD vaccinated chickens slightly increased the activity of this enzyme to some extent. There was significantly lower catalase activity in the vaccinated group C and group D. Amongst both the vaccinated groups C and D, there was non-significant lower values of catalase activity in group C as compared to group D. Spirulina supplementation in IBD vaccinated chickens slightly increases the activity of this enzyme in IBD vaccinated chickens. The glutathione reductase activity was lower in group B chickens among all the groups. Glutathione reductase activity was significantly higher in the IBDV vaccinated group C chickens. Among both the vaccinated groups (group C and D) glutathione activity was significant lower in group D chickens as compared to group C chickens. Lipid peroxidation activity (LPO) was significantly higher in group C chickens and Spirulina supplementation in IBD vaccinated chickens slightly reduces the activity of this enzyme in IBD vaccinated chickens. Gross lesion score in different organs as well as overall gross lesion score was higher in IBD vaccinated chickens. Microscopic lesions in IBD vaccinated chickens were hepatitis, inflammatory reaction around portal area in liver, congestion, haemorrhage, and tubular degeneration in kidney. Congestion and haemorrhage in lungs, intestine, thymus, spleen and caecal tonslis. Depletion of lymphocyte in bursal follicle, interfollicular inflammatory reaction and fibrosis was seen in bursa. Spirulina was able to reduce the gross and microscopic lesions score to some extent as compared to Spirulina non supplemented group chickens. Spirulina was able to provide 73.61% GLS basis and 58.40% HLS basis protection in IBDV vaccinated chickens.ThesisItem Restricted Epidemiology of enteritis in broilers and its association with α and β toxins of Clostridium perfringens(LUVAS, 2009) Agrawal, Anshul; Narang, GulshanThe present work was carried out to study the epidemiology of enteritis in broiler chickens in parts of Haryana as well as to look for its association with alpha and beta toxin of Clostridium perfringens. The data related to enteritis cases for the period from March, 08 to February, 09 were analyzed. During this period, 484 cases of enteritis were recorded in broiler chickens with overall morbidity of 3.15%. The disease was observed through out the year but maximum number of cases were recorded during winter season. Maximum cases of enteritis (158) were recorded in birds of 29-35 days of age. Maximum number of enteritis cases were associated with cocccidiosis, indicating that coccidiosis is a predisposing factor for enteritis. A total of 68 intestinal samples of poultry were screened for isolation of C. perfringens and biochemical characterization. Of the 68 samples, 49 intestinal samples (72.05%) were positive for C. perfringens. All the 49 samples gave black wool like colonies on Tryptose Sulphite Cycloserine (TSC) agar enriched with thioglycollate broth. The bacteria isolated were gram positive rod, non-motile with square ends. On TSC agar colonies were round yellowish-gray, semi-translucent and smooth with entire edge. Screening of 49 C. perfringens isolates revealed that majority of isolates (89.79%) expressed lecithinase activity and 83.60% isolates showed haemolysis on sheep blood agar. Antibiogram studies of 30 C. perfringens isolates with 11 antibiotics showed highest sensitivity to ciprofloxacin (93.3%), followed by enrofloxacin (86.6%) and moderate sensitive to chloramphenicol (76.6%), oxytetracycline (70%) and co-trimaxazol (63.3%). The organism showed 100% resistance to streptomycin and neomycin. A total of 28 C. perfringens isolates were subjected to vero cell assay to evaluate the expression of cytopathic effect (CPE). It was observed that 22 isolates (78.5%) expressed CPE in vero cell culture. Percent cytotoxicity of toxin on vero cells varied from 50.2% to 77.3% and percent dilution of toxin which showed CPE varied from 1:2 to 1:64 dilution. A total of seventy (70) intestinal fluid samples (62 from cases of necrotic enteritis and 8 cases of necrotic enteritis) were collected from dead birds and were tested by ELISA. The results revealed that five samples (7.14%) were positive for alpha toxin alone, two (2.87%) samples were positive for beta toxin alone and three (4.28%) samples were positive for both alpha and beta toxin. Out of these eight necrotic enteritis cases, four (50%) samples were positive for alpha toxin alone and three (37.5%) samples were positive for alpha and beta toxin, and one sample was negative for both of them. A total of 28 isolates of C. perfringens were tested for presence of cpa gene (alpha toxin) and cpb2 gene (beta2 toxin) by polymerase chain reaction (PCR). Number of isolates positive for alpha toxin alone were eleven (39.3%) indicates type ‘A’ Clostridium perfringens. Number of isolates positive for both alpha and beta2 toxin were nine (32.1%) indicates type ‘C’ Clostridium perfringens. Eight isolates (28.6%) were negative both for alpha and beta2 toxin indicating some other type of toxin producing Clostridium perfringens. Nucleotide sequence results for cpa gene (α toxin) revealed that three isolates of present study had 100% similarity with positive control (type ‘C’ Clostridium perfringens), whereas 98.2-100% similarity with cpa/plc gene of strains used for comparison. Nucleotide sequence results for cpb2 gene (β2 toxin) revealed that three isolates of present study had 100% similarity with positive control (type ‘C’ Clostridium perfringens).ThesisItem Restricted Molecular characterization of infectious bursal disease virus isolates from field cases in broiler chicken(LUVAS, 2009) Sunil Kumar; Narang, GulshanThis study was conducted to understand the epidemiology of infectious bursal disease (IBD) in broiler chicks in parts of Haryana as well as to characterize the infectious bursal disease virus (IBDV) by reverse transcription-polymerase chain reaction (RT-PCR), restriction enzyme analysis (RE analysis) and sequencing. The data related to IBD outbreaks for the period from July, 2005 to June, 2008 was analyzed. During this period, 483 outbreaks occurred in broiler chicks with overall morbidity, cumulative mortality and case fatality rate of 4.54, 2.34 and 51.69%, respectively. The disease was recorded through out the year and approximately 86% of the outbreaks were ii in the birds of age 21-40 days. Of these outbreaks, 334 were recorded in vaccinated flocks while 149 were in unvaccinated flocks. Per cent morbidity and cumulative mortality were comparatively higher in unvaccinated flocks than in vaccinated flocks. Detection of IBDV by RT- PCR was carried out in 30 field samples and 4 vaccine strains by amplifying the VP1 gene and hypervariable region of VP2 gene. The IBDV was detected in 28 field samples and all the 4 vaccine strains. The 432 bp PCR products of VP1 gene and 479bp PCR products of VP2 gene of 19 field isolates and 4 vaccine strains were subjected to RE analysis'. Sca1 and Pst1 restriction enzymes were used for VP1 and BstN1, Ssp1 and Pst1 were used for VP2 gene. No restriction site was observed for Pst1 and Sca1 restriction enzymes in PCR products of VP1 gene. However, in case of VP2 gene, 13 of the 19 field isolates were Ssp1+ , Pst1+ and BstN1+ , suggestive of vv types. Three fields isolates were Ssp1- , Pst1- and BstN1+ similar to classical strains. Remaining 3 field isolates were Ssp1+ , Pst1- and BstN1+ and were similar to MB and I+ vaccine strains. Phylogenetic analysis based on the nucleotide sequence of VP1 gene of four IBDVs revealed that all four were of vv type and were close to the already reported very virulent field isolates from India and abroad (KT1/99 India, OKYM, Japan, UK661, TL2004 China, 9109 USA). The nucleotide sequence analysis of VP2 gene of 10 IBDVs revealed that 9 were of vv nature due to the presence of substitutions at 795G, 827T, 833C, 857C, 897A, 905T, 908T, 1011A and1094G , specific to vvIBDV strains. Remaining one was classical/ avirulent type. There was a unique change of amino acid from alanine (A), serine (S), glutamic acid (E) to threonine (T) at amino acid position 270 in IBDVs of this study. The phylogenetic analysis suggested further divergence of IBDVs. Nucleotide and deduced amino acid sequencing revealed that the presence of vvIBDV strains in Haryana which are closely reported to the vvIBDVs from UK and Japan and also suggested continuation of changes at genetic level in these strains.ThesisItem Open Access Sequential immunopathological studies on infectious bursal disease vaccines in broiler chickens(LUVAS, 2019) Sushma; Narang, GulshanThe present study was conducted to delineate the sequential immunopathological changes in broiler chickens vaccinated with intermediate plus IBD vaccines P and Q, which are currently used in Haryana. A total of 108 day old chickens were divided into different groups viz. Group A chickens received no vaccine and served as control, chickens vaccinated with vaccine P and Q at 17 days of age were designated as group B1 and group C1, respectively while chickens which were given primary dose of vaccine at 17 days of age with vaccines P and Q, then subsequently on 24 days of age received booster dose of these vaccine were designated as group B2 and C2, respectively. No pathological lesions were observed in group A at different time interval i.e. 7, 14, 21 and 28 days post vaccination (DPV). Bursa of Fabricius (BF) showed congestion and oedema, vascular changes such as congestion and haemorrhages in other organs at 7 DPV. The severity of lesions was more evident at 14 and 21 DPV. However, lesions started to resolve at 28 DPV. Microscopically, BF from vaccinated groups revealed congestion, haemorrhages and depletion of lymphocytes. However, mild to moderate vascular changes were observed in thymus, caecal tonsils, liver, kidneys, lungs, proventriculus, intestine and heart. Overall average GLS and HLS was significantly (p≤0.05) higher in booster vaccinated group B2 (vaccine P) and group C2 (vaccine Q) as compared to single vaccinated group B1 (vaccine P) and group C1 (vaccine Q). Vaccine Q produced more severe histopathological lesions both in booster as well as in single vaccine groups as compared to vaccine P. Group D, comprised of IBD suspected field cases from 14 different poultry flocks of Haryana with most of flocks reported to be vaccinated with B2K, Georgia and intermediate plus hot strain vaccines at different ages (12-15 days of age). Grossly, petechial and ecchymotic haemorrhages were observed in pectoral, thigh and leg muscles. BF revealed oedema, fibrinous exudate and severe haemorrhages. However, other organs revealed congestion and haemorrhages. Microscopically, BF showed severe congestion, haemorrhages, marked depletion and necrosis of lymphocytes, fibrous connective tissue proliferation in inter-follicular areas along with heterophilic infiltration. Thymus, spleen and caecal tonsils revealed severe congestion, marked haemorrhages, mild depletion and necrosis of lymphocytes, while vascular changes in other organs. To ascertain the role of T cells with pathological lesion observed in BF of chickens vaccinated with P and Q vaccines as well as in IBD affected filed cases, expression of CD8+ and CD4+ T cells were studied by immunohistochemistry. Vaccine Q produced more immunohistochemical expression for CD8+ and CD4+ T cells both in single as well as in booster vaccinated group as compared to the respective groups of vaccine P. In IBD affected field cases, immunopositive CD8+T cells were diffusely present in cortico-medullary portion throughout the bursal follicles. Widely scattered CD4+T cells were present in follicles beneath bursal epithelium, inter-follicular and cortico-medullary region of BF. But the numbers of immunopositive cells were less in comparison to CD8+ T cells. The relative mRNA expression of cytokine genes i.e. IFN-γ, IL-1β and IL-6 in vaccinated chickens by real time PCR when compared with β- actin gene at different time intervals showed an up-regulation at 14 and 21 DPV in all vaccinated groups as compared to control group. From the present study, it can be concluded that both types of intermediate plus vaccine produced immunosuppression as evidenced by the different degree of lesions in BF. In single as well as booster vaccination, the vaccine Q produced more lesions in BF as compared to vaccine P. Lesions were also evidenced by upregulated expression of cytokine genes such as IFN-γ, IL-1β and IL-6 and significant increase in CD8+ T cells in the BF. These responses were more pronounced with vaccine Q in single as well as booster vaccination as compared to the respective groups of vaccine P. From the present study, it can be concluded that both types of intermediate plus vaccine produced immunosuppression as evidenced by the different degree of lesions in BF. In single as well as booster vaccination, the vaccine Q produced more lesions in BF as compared to vaccine P. Lesions were also evidenced by upregulated expression of cytokine genes such as IFN-γ, IL-1β and IL-6 and significant increase in CD8+ T cells in the BF. These responses were more pronounced with vaccine Q in single as well as booster vaccination as compared to the respective group of vaccine P.ThesisItem Open Access Studies on epidemiology of infectious bursal disease and immune response to its vaccines in broiler chicken(LUVAS, 2017) Pooja; Narang, GulshanThis study was conducted to understand the epidemiology of infectious bursal disease (IBD), to check optimal age for IBDV vaccination and immune response offered by currently available intermediate plus vaccines in broiler chicken flocks in Haryana state. The data related to IBD outbreaks for the period from January 2008 to June 2016 was analyzed. A total of 1368 outbreaks of IBD occurred during this period with overall morbidity, mortality and case fatality rate of 4.19%, 2.61% and 62.34% respectively. The disease was recorded throughout the year with maximum outbreaks in August month and in July to September quarter. Approximately 1083 outbreaks were in the 11-30 days age birds. Moreover the number of IBD outbreaks is increasing in the birds of 11-20 day of age. Of these 918 were recorded in vaccinated flocks while 450 were in unvaccinated flocks. Case fatality rate was higher in unvaccinated flocks. To check optimal age of vaccination in broiler chicken, serum samples were collected from 18 hatcheries. Maternally derived antibody titres of all the serum samples collected from different hatcheries and day of vaccination was calculated as per Deventer formula. Maternal antibody titres from the serum samples of day old chicks of 18 hatcheries showed a wide variation and age of vaccination ranged from 7-19 day of age. The non-uniformity in the MAb titres of day-old broiler chicks was expressed as percentage of coefficient of variation (CV %) with a range of 28.2% to 75.3%. Due to wide range of MAb titre, twice vaccination is advisable. As per our study however, calculating the age of vaccination for the chicks from individual hatchery is advisable if possible. To evaluate the vaccine immune response of IBD vaccines, an experimental study was conducted using two commercial intermediate plus vaccines P and Q. The level of MAbs ranged from 6834.6 to 9925.60 in day old chicks. In experimental group age of vaccination came 17 because maternal antibody titre was uniform and high. The mean maternal antibody titre of chicks at 1 day age, 7 and 14 day age was 7874.21, 2449.4 and 1089.9. The mean MAb titre shows a steady decrease in the titre from day old age to 14th day of age. Both vaccines P and Q provided protective immunity from 7 DPV till the end of experiment. Vaccine P showed static increase in ELISA antibody titres from 7 DPV however vaccine Q showed quite higher level of antibody titres on 7 DPV itself which remain protective till the end of experiment in both. CD4+ cells increased at 7 DPV, decreased at 14 and 21 DPV, again increased at 28 DPV. CD8+ cells increased at 7 DPV, decreased at 14 DPV, again increased at 21 and 28 DPV. This could probably be due to immunosuppressive effect of IBD vaccines. Bursal index was comparable with control in case of birds vaccinated with vaccine P at 7 DPV only. However it was reduced significantly in birds vaccinated with vaccine Q at 7 DPV itself. Later the bursal index was significantly lower in all the vaccinated groups vaccinated with either of the vaccine at 14, 21, 28 DPV immunosuppression. Histopathology showed lymphoid depletion, medullary necrosis with cyst formation, fibrous connective tissue proliferation in vaccinated groups which developed early and was comparatively more with vaccine Q.ThesisItem Restricted Studies on pesticide residues in poultry feed, water and meat(LUVAS, 2015) Mishra, Anil Chandra; Narang, GulshanBroiler poultry birds intended for human food get exposed to pesticides through feed, water and environment. These residues get accumulated in the muscle, adipose and organ tissues of exposed birds. Therefore the objective of the present investigation was to study the residues of organochlorine (OC), pyrethroids and organophosphorous (OP) pesticide residues in poultry feed, water and meat at six selected broiler farms of Hisar along with 50, random meat samples. A small survey on pesticide residues in pond water samples from six districts of Haryana was also conducted. Multiresidue methods were standardized for simultaneous extraction and detection of residues of OC and pyrethroid pesticides viz. α-HCH, β-HCH, γ-HCH, δ-HCH, α-endosulfan, β- endosulfan, endosulfan sulfate, λ-cyhalothrin, cypermethrin and deltamethrin by GC-ECD technique and residues of OP pesticides i.e. dichlorovas, monocrotophos, pirimiphos methyl, fenitrothion, malathion, chlorpyriphos, quinalphos and edifenphos by GC-NPD technique. Sample matrices of poultry feed and meat were processed as per the method described by Dutch ministry of Public Health, Welfare and Sports, the Netherlands (AMPRF, 1996), whereas water samples were processed as per the method of Kouzayha et al. (2012). The results of six selected farms revealed that only one sample each of feed and water out of 18 was positive for OC, where in the concentration levels of total HCH was3.914μg/kg, total endosulfan 4.346 μg/kg, while those of endosulfan in poultry water was 0.103μg/kg and meat was not detected with any of the OC’s. At these residue levels, meat of poultry birds from same farm did not showed detectable level of OC pesticides. Pyrethroids were also not detected in any of the sample matrices. The mean levels of monocrotophos, pirimiphos methyl, fenitrothion, malathion, chlorpyriphos and quinalphos in poultry feed were 7.553, 2.257, 3.622, 9.742, 2.760 and 15.792 μg/kg respectively. Similarly in water sample, monocrotophos, pirimiphos methyl, fenitrothion and chlorpyriphos were detected in only one sample in the concentrations 0.0454, 0.083, 0.054 and 0.057 ηg/ml respectively. While the mean levels of monocrotophos, pirimiphos methyl, fenitrothion, malathion, chlorpyriphos and quinalphos were 14.352, 6.73, 15.729, 3.63, 12.38 and 6.084 μg/kg respectively in meat. The results of random meat samples (50) revealed the presence of HCH in 9 samples and endosulfan in 5 samples, while pyrethroids were not detected in any of the samples. In case of OPs, the monocrotophos was found in maximum 8 samples (16%), followed by chlorpyriphos in 5 samples (10%), dichlorovas and malathion in 3 samples (6%) each and qunalphos in 1 sample (2%).Comparison of pesticide concentration in each positive sample of meat with national and international MRLs showed that, endosulfan and lindane among OC and chlorpyriphos among OP pesticides were responsible for maximum violations. The results of present study demonstrated that the poultry feed and water is significance source of pesticide residues in poultry meat. In case of pond water samples OC and Pyrethroids were not detected in any of the sample however OPs were detected in 16 samples with 32% of occurrence rate. Among the OPs monocrotophos and chlorpyriphos were found in maximum of 8 samples (16%) each, followed by pirimiphos methyl in 3 (6%), malathion in 2 (4%) and fenitrothion, quinalphos and edifenphos in 1 sample (2%) each.