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Lala Lajpat Rai University of Veterinary & Animal Sciences, Hisar
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ThesisItem Open Access Development of an indirect ELISA and study of immune response to Clostridium chauvoei in cattle(LUVAS Hisar, 2023-08-17) Tejaswi; Gupta, Akhil KumarIn the present study, an Indirect ELISA was developed to demonstrate antibodies against Black quarter (B.Q.) vaccine antigen. The B.Q. vaccine antigen was retrieved from the vaccine formulation by dialysis to remove the formalin and was used as antigen. The hyper immune serum raised in rabbits against B.Q. vaccine antigen was used as positive control in ELISA development. The optimized dilutions of the reagent used in ELISA were 5 µg/ml for coating antigen; 1:200 dilution for hyper-immune rabbit serum; 1:200 dilution for pre-immune Rabbit serum; 1:200 dilution for Cattle serum and 1:8000 dilution for Protein G HRP conjugate. The cut-off value (mean+3SD) for the developed ELISA calculated using the naive calf serum (n=37) was found to be 0.347135. In this study, the animals kept at Animal Farm, Department of Animal Genetics and Breeding were divided into three groups with 10 animals in each group: Group I (calves; < 9- month age), Group II (heifers; 18-to-26-month age), and Group III (adults; > 26-months age). In Group I 70% of the calves; in Group II 90% of the heifers and in Group III 100% of the adults showed one month post vaccination immune response (in terms of OD450) above cut-off value. However, the kinetics of the immune response studied over a period of six months was found to be aberrant which required to be further investigated. Based on the above study, we can conclude that some preliminary base had been developed and further improvement and refinement is required in the ELISA so that we can calculate sero-conversion in vaccinated animals in terms of antibody titerThesisItem Open Access Modification of foxnut starch and its application in low fat Yogurt(LUVAS Hisar, 2023-09) Anuj; Arora,ShaliniThe demand for dairy products is steadily increasing worldwide. Yogurt, being a healthy beverage, is preferred by consumers but due to health reasons, desires lower fat content in the product. However, reducing the fat content in yogurt can result in a loss of its original texture and consistency. To address this issue, texturizing agent could be added to produce low-fat yogurt. In this study, we aimed to develop foxnut starch based novel texturizing agent by physical modification. Further, the developed product was evaluated of its effectiveness in low-fat yogurt. Two different physical modification approaches, namely heat moisture treatment (HMT) at moisture levels of 20%, 25%, and 30% for 4 and 8 hours, respectively, at 110ºC, and annealing at 50ºC and 55ºC for 6 and 12 hours, respectively, were employed. The modified starch samples were analysed for their functional, colour, and pasting properties. Based on the desired functionality and pasting properties, the HMT modified starch treated at 20% moisture for 4 hours and the annealing modified starch at 55ºC for 6 hours were selected for application in low-fat yogurt. The native foxnut starch exhibited a swelling power of 2.98g/g, solubility of 9.28%, oil absorption capacity (OAC) of 0.32g/g, emulsifying capacity (EC) of 89.39%, and water absorption capacity (WAC) of 1.91g/g. The pasting temperature and oil absorption capacity showed a significant increase (p ≤ 0.05) after the HMT and annealing processes. On the other hand, starch solubility and swelling power decrease significantly (p≤0.05) with increasing moisture levels, temperature, and treatment duration during HMT and annealing. The addition of modified starch significantly reduced syneresis in low-fat yogurt from 11.55% (control) to 8.28%, while water holding capacity increased from 33.4% (control) to 39.11% at a 2.0% modified starch level. Starch contributed to water retention, decreased syneresis, and enhanced mouth feel, colour, and body of the yogurt. The optimal concentration of modified starch for improving the texture, body, and overall acceptability of yogurt with reduced syneresis was found to be up to 1%. Beyond this limit (starch concentration above 1%), it resulted in undesirable sensory attributes such as chalkiness, lumpiness, reduced tongue spread ability, and excessive hardness.ThesisItem Open Access Development of ice cream incorporated with Spirulina platensis and inulin(LUVAS Hisar, 2023-06) Rathore,Jyotsana; .Monika RaniThe study was conducted to develop a low sugar functional ice cream by incorporating Spirulina platensis and inulin at various levels individually as well as in combination along with other ingredients. On the basis of sensory evaluation, out of twenty-five treatments (four variations with Spirulina platensis, four with inulin, sixteen combinations of both including control), one treatment was selected from each group, namely S2 from Spirulina platensis ice cream, I4 from inulin ice cream, and S2I4 from the combination group. The selected treatments underwent further analysis for proximate composition, physicochemical parameters, whipping ability, melting rate, overrun, antioxidant activity, and total phenolic content. The ice cream formulations I4 and S2I4 showed 3 % lower sugar, while S2 and S2I4 were having 17.89% higher protein, and 9.6 times higher iron content. Moreover, the formulation (S2 and S2I4) exhibits 2 times higher antioxidant activity. The Spirulina platensis and inulin incorporation improved whipping ability and increased overrun values, leading to a longer meltdown time, thus enhancing the ice cream's stability during consumption. Further the selected treatment (S2I4) was further studied for storage at freezing temperature (-18 to -20oC). The physicochemical properties of the S2I4 treatment showed lower pH and higher titrable acidity, which changed during storage but remained within acceptable limits and the presence of Spirulina platensis powder resulted in a darker color and a greener hue, while inulin had no significant impact on color. On organoleptic evaluation, the final treatment (S2I4) maintained its high sensory scores for up to 30 days, making it safe for consumption. Microbiological analysis indicated that the microbial counts (Standard plate count, coliform count, psychrotrophic count) of developed ice cream were within safety limits defined by FSSAI. Furthermore, cost economics study revealed a slightly higher cost of the developed S2I4 ice cream compared to the control, but the superior nutritional and sensory attributes justified the difference.ThesisItem Open Access Development of reverse transcription recombinase polymerase amplification assay for rapid detection of porcine sapelovirus(LUVAS Hisar, 2023-04) Kaur, Ramandeep; Sushila MaanPorcine Sapelovirus (PSV) A, a member of newly classified genus Sapelovirus, belongs to family Picornaviridae. PSV is non-enveloped virus with icosahedral symmetry with diameter of around ~35nm and the genome is a linear, non-segmented, single-stranded positive-sense RNA. PSV infection in pigs is mostly asymptomatic, but it can also cause severe neurologic, enteric, and respiratory symptoms or reproductive failure. Sapelovirus infections have been reported worldwide in pigs. PSV is considered as the emerging pathogen of porcine species. For the virus detection, isothermal Recombinase polymerase amplification (RPA) were developed. The RT-RPA assay was developed using commercially synthesized positive control and primers based on 5´conserved region of PSV genome. The assay was optimized at low and constant temperature of 35°C for 20 minutes with forward primer concentration 20pm, reverse primer concentration 5pm and MgOAc concentration of 14mM. The analytical sensitivity of assay was determined using serial dilutions of positive control. The limit of detection was up to 283 copies using agarose gel electrophoresis. The assay was specific to PSV and no cross reactivity was observed with other related viruses. The assay was validated using field samples followed by visual assay for detection. A total 85 samples were collected from different organized and unorganized piggery farms of Haryana. Out of 85 samples 21 samples were found positive for PSV. The positivity rate of PSV was 24.7%. This is the first report of PSV from Haryana suggesting the virus circulation among pig population. Therefore, the above study concludes that the developed RT-RPA assay is rapid, highly sensitive and specific and works at low, constant temperature and do not require any special instrument. Visual dye-based detection makes the developed RPA assay a potential candidate for pointof-care testing (POCT).ThesisItem Open Access Effect of pellet diets with or without phytogenic feed additives on performance of broilers(LUVAS Hisar, 2023-04) Devika; Sihag,Zile SinghA trial of 42 days was carried out to study the effect of pellet diets with or without phytogenic feed additives on performance of broilers. A total of 240, day-old commercial broiler chicks were randomly distributed into eight dietary treatments with three replicates per treatment and each replicate had ten birds. Ashwagandha and shatavari were used as herbal feed additives. The basal diet was formulated as per BIS (2007) specifications. The treatments included T1: Basal diet + antibiotics (mash form), T2: Basal diet + antibiotics (pellet form), T3: Basal diet + 1% Ashwagandha powder (mash form), T4: Basal diet + 0.5% Ashwagandha powder (pellet form), T5: Basal diet + 1% Ashwagandha powder (pellet form), T6: Basal diet + 1% Shatavari powder (mash form), T7: Basal diet + 0.5% Shatavari powder (pellet form) and T8: Basal diet + 1% Shatavari powder (pellet form). Body weight (g/bird), Feed intake (g/bird) and FCR improved significantly (P<0.05) after supplementing the diet with phytogenic feed additives and the best results were observed in 1% Ashwagandha supplemented group in pellet form. In between the control groups, the pelleted diet fed group had better growth performance than mash diet fed group. Dietary supplementation of 1% Ashwagandha in pellet form resulted in significantly (P<0.05) higher DM metabolizability and gross energy metabolizability as compared to control. Nitrogen retention was found significantly (P<0.05) higher in groups T3, T5, T6 and T8. The carcass traits were found to be significantly (P<0.05) better in 1% Ashwagandha supplemented group in pellet form. The Hb levels and TLC count were found to be significantly (P<0.05) higher in all Shatavari supplemented groups while decreased % of heterophils and increased % of lymphocytes thereby, reducing heterophil: lymphocyte ratio in T6, T7 and T8 groups compared to control. No significant effect of pelleting was observed on haematological parameters. Significant reduction in serum cholesterol and triglycerides levels was observed in 1% Ashwagandha and 1% Shatavari supplemented groups. Inclusion of 0.5% and 1% Ashwagandha and Shatavari powder resulted in significant improvement in HDL and reduction in LDL as compared with control. But no effect of pelleting was observed on serum parameters. A significant reduction in E.colicount was recorded in the T3 and T5 groups compared to control. Higher values of Lactobacilli were observed in all herbal additives supplemented groups. Highest villi height andVillus height: Villi depth of intestinal crypts was recorded in 1% Ashwagandha supplemented group in pellet form. The activity of antioxidant enzymes; SOD and Catalase was observed in groups T3, T5, T6 and T8. Thus, the dietary supplementation of Ashwagandha root powder and Shatavari root powder leads to significant improvement in the growth performance, nutrient metabolizability and hematological indices; possess potent hypocholestrolemic, and gut microbiota stabilizing effects, thus help in achieving a profitable and feasible poultry production. The feeding of pelleted ration also improved the growth performance of birds and this will help in establishing profitable poultry farming.ThesisItem Open Access Comparative evaluation of interlocking nailing alone and in combination with Beta–Tricalcium Phosphate (β-TCP) for management of long bone fracture in dogs(LUVAS Hisar, 2023-04) Manikant; Sandeep SaharanThe objective of this research was to determine whether beta-tricalcium phosphate (β-TCP) might be used to enhance fracture healing following interlocking nail stabilisation. Twelve dogs with long bone fractures, regardless of age, breed, or sex, were divided into two groups at random. Six randomly chosen animals from group I underwent interlocking nailing, whereas six animals from group II underwent interlocking nailing as well as the application of β-TCP to the fracture site. On the day of presentation, a detailed clinical history was taken, and radiographic, haematological, biochemical, and clinical examinations were performed before surgery as well as on the 15th, 30th, and 60th postoperative days. At different intervals, group II performed better than group I in terms of weight bearing when walking and standing. The higher bone union and bone formation scores (BFS and BUS, respectively) were seen in group II compared to group I. In group II, the stage of bone union and functional limb usage was achieved earlier as compare to group I. The haemato-biochemical and physiological parameters did not differ significantly between the groups. Also, the complications identified in this analysis included suture dehiscence, delayed healing, screw loosening, and suture tract infection. According to the findings of the current study, interlocking nailing combined with the application of βTCP was found to be more effective than interlocking nailing alone in the management of long bone fracture in dogsThesisItem Open Access Effect of inclusion of Moringa leaf meal and pelleting of the ration on the performance of broilers(LUVAS Hisar, 2023-04-10) Shabnam; Tewatia,B.S.An experiment was conducted to study the effects of pelleting and Moringa oleifera leaf meal supplementation on the performance of broiler chicken. One hundred sixty, one day old chicks, were randomly divided into 4 treatment groups having 4 replicates each, and each replicate had 10 birds. The control group (T1) was offered basal diet with antibiotics in mash form; T2 group was fed basal diet with antibiotics in pellet form; while birds in T3 group were offered ration having 5% Moringa oleifera leaf meal (MOLM) without antibiotics in mash form and T4 group ration had 5% Moringa oleifera leaf meal without antibiotics in pellet form. During the feeding trial of 42 days, feed intake and body weight changes were recorded. At the end of feeding trial, a metabolism trial of five days duration was conducted during which samples of feed offered, residue left and faeces voided were collected. For carcass evaluations, one bird from each dietary treatment was sacrificed. The average feed intake (g/bird) during overall period (0 to 42 day) did not differ significantly among groups. Average body weight gain (g/bird) in all the three growth phases differed significantly (P<0.05) and maximum body weight gain (2281.32) was recorded in group (T4) supplemented with 5% Moringa oleifera leaf meal in pellet form. Similar trend was observed in respect of feed conversion ratio, carcass traits, DM metabolizability, nitrogen metabolizability and gross energy metabolizability. Quality of the breast and thigh muscles in terms of crude protein (21.97, 17.11) and ether extract (5.52, 6.76) also improved significantly (P<0.05) in T4 group. Hematological parameters (Hb, TEC and lymphocyte) increased significantly (P<0.05), while TLC and heterophils decreased (P<0.05) in T4, T3 and T2 as compared to the control group. A significant (P<0.05) reduction in serum total cholesterol, LDL and triglycerides levels and increase in HDL was also observed in Moringa supplemented groups (T4 and T3) as compared to the antibiotic supplemented groups (T1 and T2). MOLM supplemented groups (T4 and T3) resulted in significant (P<0.05) decrease in E.coli count and increase in Lactobacilli count as compared to T1 and T2 groups. Antioxidant enzymes (SOD and Catalase) activity was also significantly higher (P<0.05) in MOLM supplemented group in pelleted form (T4) followed by T3, T2 and T1 group. Villus height: intestinal crypts depth of the experimental birds ranged from 4.47µm (T1) to 6.30 µm (T4) and significantly (P<0.05) higher effect was observes in the groups supplemented with MOLM. Thus, it can be inferred from the results of this study that inclusion of 5% MOLM in pellet form had better effect on body weight gain, FCR, carcass traits and antioxidant enzyme activity besides being better in terms of feeding cost per kg gain in broilers.ThesisItem Open Access Effect of feeding pelleted ration with or without herbal feed additives on the performance of broiler chickens(LUVAS Hisar, 2023-04) Radha Bai; R.S. BerwalThis research was conducted to investigate the effect of feeding pelleted ration with or without herbal feed additives on the growth performance, nutrient utilization, carcass traits, hematological, serological parameters and to economise the feeding of broiler chickens. Study was conducted on two hundred and forty, day-old broiler chicks which were randomly divided into eight dietary treatment groups with three replicates of each treatment. Each treatment had different feeding regimen: T1 received a basal diet with antibiotics in mash form; T2: basal diet with antibiotics in pellet form; T3: basal diet with 1% aloe vera in mash, T4: basal diet with 0.5% aloe vera in pellet form, T5: basal diet with 1% aloe vera in pellet form, T6: basal diet with 1% amla in mash form, T7: basal diet with 0.5% amla in pellet form and T8: basal diet with 1% amla in pellet form. In the 42-day feeding trial, Feed intake and body weight were recorded at every week and residual feed and faecal samples were collected and analyzed during a five-day metabolic trial during the end of growth trial. The results showed that the average overall body weight differ significantly (p<0.05) in amla and aloe vera supplemented groups in comparison to control groups (T1 and T2 ), with the highest average body weight was recorded in the group supplemented with 1% amla in pellet form, followed by 1% amla in mash form. Better feed conversion ratio was observed in 1% aloe- vera supplemented group in pellet form (T5 ) followed by T8 , T4 , and T3 . Significantly (p<0.05) higher nutrient metabolizability was observed in 1% aloe vera supplemented group followed by 1% amla supplemented groups. Significantly (p<0.05) higher percentage of carcass traits were observed in 1% amla supplemented groups in pellet and mash form as compared to the control groups. Hematological parameters (Hb, TEC, TLC and lymphocyte) were significantly (p<0.05) improved, while heterophils were significantly (p<0.05) decreased in the 1% amla supplemented groups compared to the control group. The herbal feed supplemented groups also showed a significant reduction in serum cholesterol, LDL, and triglyceride levels, as well as an increase in HDL levels, compared to the antibiotic-added groups. But better results were obtained in 1% aloe vera supplemented groups. The groups supplemented with 1% aloe vera in pellet and mash form had a significant decrease in E. coli count and an increase in Lactobacilli count compared to the T1 and T2 groups. The antioxidant activity of SOD and catalase enzyme were significantly (p<0.05) increased in all herbal feed supplemented groups with the highest improvement observed in T8 followed by T6 , T5 , and T3 . The villus height and intestinal crypts depth of the experimental birds ranged from 898.41µm (T1 ) to 1035.40 µm (T5 ) and 3.15 µm (T1 ) to 4.24 µm (T5 ) under different dietary treatments, and herbal feed supplemented group had significantly better effect on gut morphometry higher villus height was observed in the groups with inclusion of 1% aloe-vera in pellet as well in mash form. Better economics was found in 1% amla supplemented group in pellet form as compared to that of other herbal feed supplemented and control groups. In conclusion, the study suggested that inclusion of amla and aloe-vera supplementation at 1% level improved the growth performance, nutrient digestibility, carcass traits, hematological and serological parameters of broiler chickens.ThesisItem Open Access Studies on the ameliorating effect of Eugenol (clove essential oil) on Aflatoxicosis in broiler chicken(LUVAS Hisar, 2023-07) Nehra,Neetu; DeepikaAn experimental study was conducted on 132 broiler chicks to assess the hepatoprotective and antioxidative potential of eugenol (clove essential oil) in broiler chicken and to evaluate the ameliorative effect of eugenol supplementation on haemato-biochemical, oxidative stress, pathological changes and immune response in aflatoxin administered broiler chicken. Aspergillus parasiticus MTCC 411 spores were revived on potato dextrose agar and Cezpak yeast extract agar and used for aflatoxin production in maize and final experimental feed was estimated to contain 184 ppb of aflatoxin. One hundred and thirty-two (132) broiler chicks at an age of 7 days were randomly divided into 2 groups viz, Group A and Group B having 66 chicks in each group. All chicks were vaccinated against NCDV as per standard schedule. Group A chicks were supplemented with eugenol (@ 5ml/kg) of feed throughout the experiment. Group B chicks were kept as control and fed with basal control diet (without eugenol). At the age of 14 days, the chicks of groups A and B were further divided into two subgroups {group A into A1(AF+E) and A2(E); group B into B1(AF) and B2(C)} of 33 chicks each. All the chicks of group A1 and B1 were given aflatoxin (AF) @184ppb mixed with feed till the end of experiment and chicks of group B2 were fed on basal feed and act as control i.e., without any treatment. Blood samples were collected from six chicks of each subgroup directly from heart on 0, 7th, 14th 21st and 28th day post aflatoxin administration (DPAA) for assessing different parameters like haematological and oxidative stress parameters in blood while biochemical parameters and humoral immune response in serum. After collection of blood, chicks were sacrificed and detailed postmortem examination was conducted. Representative tissues of the different organs were preserved with 10 per cent neutral buffered formalin for histopathological examination. The aflatoxin administered chicks (group B1 and A1) exhibited varying degree of clinical signs such as dullness, ruffled feathers and reduced feed intake at different time intervals. Mean body weight was significantly decreased in groups B1 and A1 as compared to control group (B2) at different time intervals. Haematological studies showed significant decrease in mean Hb, PCV and TEC and non-significant decrease in MCV and MCHC in groups B1 and A1 at different time intervals. Mean TLC was also found significantly decreased in group B1 and A1. Biochemical studies showed significant decrease in mean total serum protein and albumin concentration and significant increase in liver function parameters like serum AST, ALT, GGT in aflatoxin fed groups (B1 and A1) at different time interval as compared to control group B2. Kidney function parameters like uric acid and creatinine concentration were also significantly increased in group B1 and A1. Blood catalase and SOD activities were significantly decreased and blood LPO (nMDA) was significantly increased in toxin fed groups as compared to control group at different time intervals. Eugenol supplementation significantly improved the body weight and ameliorated the adverse effect of aflatoxin on hemato-biochemical parameters and oxidative stress parameters. Immunological studies revealed significant decrease in humoral immune response in aflatoxin administered groups as observed by reduced mean HI titre (log2) against NDV in Group B1 and A1. Delayed type hypersensitivity study to assess CMI also revealed significant decreased skin thickness 24 hour after DNCB application in aflatoxin administered group B1(AF) as compared to control groups B2(C) and A2(E) while slight reduction with no significant difference was observed in group A1(AF+E). Pathomorphological lesions characteristic of aflatoxicosis were prominent in liver, kidneys and lymphoid organs and eugenol supplementation reduced the severity of lesions. In case of histopathology the lesions were vacuolar, vascular, fatty, degenerative, necrotic, infiltrative and proliferative in aflatoxin intoxicated group in different organs and were of mild nature in eugenol supplemented group A1(AF+E). Results of the present study indicated the hepatoprotective and antioxidative potential of eugenol (@ 0.5% in feed) along with its ameliorative effects against aflatoxin induced pathological alterations in broiler chicken
