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  • ThesisItemRestricted
    Detection and molecular characterization of foot and mouth disease virus from persistently infected bovine
    (2023-07) Ankit Pannu; Dahiya, Swati
    Foot-and-mouth disease (FMD) is an OIE-listed highly contagious and economically devastating disease of cloven-hoofed animals. Routinely, FMD virus (FMDV) 3AB3 non-structural protein (NSP) based indirect ELISA test is used for population serosurveys as an indicator of exposure to virus to differentiate between infected and vaccinated animals (DIVA) in India. For this, serum samples (n=377) collected from 6-18 month old cattle and buffaloes from nine villages of district Hisar, Haryana during 2022 as per the sample plan developed by ICAR-NIVEDI were subjected to DIVA ELISA. NSP reactivity was found to be 5.3% (20/377) in both, cattle (11.3%; 12/106) and buffaloes (3.0%; 8/271). Two buffaloes from 20 NSP reactors when sampled at a gap of nine months were found to be negative for FMDV 3AB3 NSP Abs as well as for FMDV by real time RT-PCR and RT- Multiplex PCR (RT-mPCR). No FMD outbreak was reported from Haryana during 2022. Further sampling was carried out from an organized cattle farm in which bi-annual FMD+HS combined vaccination has been carried out regularly. There was no history of FMD outbreak on this farm for more than a decade. DIVA reactivity of 43.75% (7/16) was observed in cattle (6-18 months age) of this farm which could be linked to either false positive reactions or scars of past exposure and virus elimination at the time of OPF sampling. The blood parameters between NSP positive and NSP negative animals were statistically insignificant. The only animal exhibiting antibody titres
  • ThesisItemRestricted
    Studies on etiology, haematobiochemical changes and therapeutics of diarrhoea in bovines
    (LUVAS,Hisar, 2023-04) Pradeep Kumar; Tarun Kumar
    Chronic diarrhoea is a significant problem in the livestock industry, leading to economic losses and requires veterinary attention. A study was conducted to understand the clinical features and hematological and biochemical changes in animals affected by chronic diarrhea. The study included detailed history of all affected animals, with an age range of 2 months to 14 years, and majority of affected animals (61.25%) were more than five years old in their third to fourth lactation (43.75%). The duration of illness ranged from 1 day to 1.5 years, and 61.25% of cases were chronic. Clinical signs included normal feed intake in 62.5% of animals, anorexia in 25%, and inappetence in 12.5%. Water intake was reduced in 83.75% of animals, and frequency of defecation ranged from 6-10 times daily in 57.5% of animals while more than 10 times daily was present in 27.5% of cases. Hematochezia and melena was found in 1.25% and 3.75% of cases, respectively. Milk production was reduced remarkably (more than 50%) in 60% of cases, causing great economic loss to the animal owners. Hematological findings showed anemia, lower hemoglobin, higher TLC, neutrophilia, and lymphopenia. Biochemical studies revealed hyponatremia, hypokalemia, hyperchloremia, hypocalcemia, hypophosphatemia, hypomagnesemia, increased activity of liver/skeletal enzymes, hypoglycemia, increased creatinine and BUN, hyperproteinemia, hypoalbuminemia, and hyperglobulinemia. These findings can aid in the diagnosis and management of chronic diarrhea in livestock. Etiological findings through highlighted the importance of locating the persistently infected animal as it will continue to spread infections to the healthy animals living in vicinity. Antibiotic sensitivity tests highlighted Amoxicillin/sulbactum as best drug. Therapeutic trial with same antibiotic further supplemented their outcome in combination with Lugol‘s Iodine. Diarrhoea as a disease condition caused by multiple etological agents.In present study occurence of BVDV as a reason for diarrhoea in bovines was reported as 2.50%, while Mycobacteruium paratuberculosis caused diarrhoea in 3.75% cases. Parasitic causes of diarrhoea were reported as 11.25%. This study highlights the significance of clinical, hematobiochemical and etiological factors associated with chronic diarrhoea.
  • ThesisItemOpen Access
    Effect of prebiotic and probiotic supplementation on performance of rabbits
    (LUVAS,Hisar, 2023) Rajan Papneja; Singh, Sajjan
    Eighteen New Zealand White rabbits of either sex at 24 weeks of age were randomly allocated to three treatments (six rabbits in each treatment). Rabbits in each treatment received different diets [Standard control diet (T1), Diet supplemented with Prebiotic @400 mg/kg concentrate (T2) and Diet supplemented with Probiotic @ 1 billion CFU/Kg concentrate (T3)] in addition to ad lib. berseem fodder. The concentrate mixture of group T1 were formulated using maize, soybean meal, ground nut cake, gram, mineral mixture and common salt. The concentrate diets formulated were isonitrogenous and iso-caloric. Results showed that diet supplemented with prebiotic and probiotic had improved growth performance parameters (body measurements) as compared to the control group. The total weight gain was statistically (p<0.05) higher in rabbits raised on diets supplemented with prebiotic (1563.67g) and probiotic (1557.17 g) as compared to standard control diet (1419.17 g). Significantly (p<0.05) higher serum total protein and globulin values and significantly (p<0.05) lower total serum cholesterol and low density lipoprotein values were observed in prebiotic and probiotic supplemented groups than un-supplemented groups. In prebiotic (T2) and probiotic (T3) supplemented groups, the comparative calculated dressing percentage (57.28 and 57.07 %), body cut up parts i.e., foreleg (135.76 and 137.06 g), hind leg (230.69 and 233.11g), back (263.88 and 270.73g) and chest (219.78 and 226.75g) and meat bone ratio (5.35 and 5.38) showed the higher values as compare to dressing percentage (53.89%), body cut up parts i.e., foreleg ( 127.46g), hind leg (217.48g), back (243.29g) and chest (209.24g) and meat bone ratio (4.91) of control group. However, the per cent moisture content showed a non-significant (p ≤0.05) difference among all the groups, but significantly (P≤0.05) higher mean per cent protein of meat was recorded in T2 (19.17%) and T3 (19.52%) groups as compared to meat of T1 (17.61%) group. The total meat cholesterol levels (mg/g) in T2 and T3 group (160.38 and 159.81 mg/g) found lower as compared to control T1 group (163.67) of rabbit meat samples. Nutrient intake, digestibility of nutrients and nutritive value of ration was significantly (p<0.05) improved in rabbits raised with prebiotic and probiotic supplementation in their concentrates. The results inferred that rabbits fed prebiotic and pro biotic supplemented diets had improved feed intake, FCR and carcass characteristics. Feed cost per kg weight gain was found lowest in treatment group which was supplemented with probiotic.
  • ThesisItemOpen Access
    Performace of Soymilk as partial milk replacer in Murrah buffalo calves
    (LUVAS,Hisar, 2023) Mohammed Ishan Hashmi
    The present study attempts to analyze the performance of Soymilk as partial milk replacer in Murrah buffalo calves. After the period of colostrum feeding at the age of 5 days, 15 Murrah buffalo calves were randomly divided into three treatment groups, having 5 buffalo calves in each treatment on the basis of similar body weight and sex. The treatments included: T1 (Whole milk feeding), T2 (Whole milk and Soymilk in 75:25 ratio) and T3 (Whole milk and Soymilk in 50:50 ratio). This experiment was conducted for a period of 90 days. Soypowder for Soymilk was prepared by soaking, dehulling, drying, grinding and sieving of soybean. For preparation of 1 litre Soymilk, 150 g soypowder was dissolved in water with continuous stirring and then strained with fine muslin cloth. Thereafter, this Soymilk was mixed with the Whole milk as per the treatment groups. The calves were given balanced diet in form of green fodder, dry roughage and concentrate mixture to meet their dietary requirements for growth as per ICAR 2013 feeding standards. The quantity of different feeds given to calves of each group was adjusted at 15 days intervals so that the overall dietary requirements of buffalo calves were met according to the change in their body weight. Body weight and body measurements (body height, heart girth, abdominal girth and body length) were recorded at the start of experiment and thereafter at 15 days interval. The feed intake (fresh basis, dry matter basis and DMI/100 kg body weight), FCR, FCE and faecal score were recorded at 15 days interval up to 90 days of experiment. Haemato-biochemical parameters were observed at monthly interval. The results of the experiment revealed that body weight parameters (average body weight, body weight gain, cumulative body weight gain and average body weight gain) and body measurement parameters (body height, cumulative body height, heart girth, cumulative heart girth, abdominal girth, cumulative abdominal girth, body length and cumulative body length) did not differ significantly due to partial replacement of Whole milk with Soymilk at 25% and 50% level during whole experimental period. Similarly, feed intake (dry roughage, green fodder and concentrate intake), FCR and FCE showed no significant difference among T1 (Whole milk feeding), T2 (Whole milk and Soymilk in 75:25 ratio) and T3 (Whole milk and Soymilk in 50:50 ratio) treatment group. Various haematological parameters (Hb, TEC, TLC, lymphocytes, monocytes, neutrophils, eosinophils, basophils, MCH, MCHC and PCV) and serum biochemical parameters (creatinine, glucose, total protein, albumin, cholesterol, calcium, chloride, SGPT, SGOT, ALP, phosphorus, urea, HDL and LDL) were no significantly different among treatment groups in which calves fed Whole milk and combination of Whole milk and Soymilk. The results also revealed that feeding of Soymilk had no effect on faecal score in experimental calves. The cost of feeding calculated at the end of experiment revealed that net profit in total feeding cost/kg weight gain were Rs. 106.92 and 130.35 in T2 (Whole milk and Soymilk in 75:25 ratio) and T3 (Whole milk and Soymilk in 50:50 ratio) treatment group with respect to T1 (Whole milk feeding) treatment group. Through this experimentation, it can be concluded that Whole milk can be partially replaced by Soymilk up to 50% level without any adverse effect on performance of Murrah buffalo calves and Soymilk feeding is more economical than Whole milk feeding.
  • ThesisItemOpen Access
    Detection of cyromazine and melamine residues in poultry eggs
    (2023-02) Soni,Deepak; Vijay J. Jadhav
    Cyromazine (CYR) is an insect growth regulator for fly control at animal farms. It is used as a feed additive in layer farms so that it gets excreted in the droppings of the birds and exert its larvicidal activity in the manure. However, due to this practice, residues of CYR and its metabolite melamine (MEL) also appear in the eggs. A new instrumental method was developed for separation and detection of CYR and MEL on HPLC-UV system. The mobile phase comprised of ACN and 0.05% TFA at pH 2.5 was applied in gradient mode at a flow rate of 1 mL/min. A new method using anhydrous magnesium sulfate, calcium silicate, celite 545 as adsorbents and acidified acetonitrile as extraction solvent was developed for extraction and clean up of CYR and MEL residues in eggs which was found to be quick, easy, cheap, effective, rugged and safe (QuEChERS). The LOD and LOQ of the method was found to be 5.3 and 16.0 μg/kg for CYR and 11.4 and 34.7 μg/kg for MEL, respectively. The accuracy of the method ranged between 84-103 % and precision of the method remained less than 5 % RSD. Linearity studies of spiked sample showed the consistent performance of the method at various concentrations (R²>0.996). A total of 110 egg samples were collected from 52 farms located in 5 different districts of Haryana (Panchkula, Ambala, Panipat, Sirsa and Hisar) and analyzed for the presence of CYR and MEL by applying newly developed QuEChERS method. The mean concentration of the CYR was found to be 42.71 μg/kg with a range of 16.88 -112.61 μg/kg. MEL was not detected in any of the sample analyzed. Estimates of dietary exposure of CYR through consumption of eggs for population in Haryana was found to be 33.40 μg/kg b.w./day which contributed 0.55 % of ADI of CYR. A cross-sectional study was carried out in 52 layer farms situated in 5 districts (Panchkula, Ambala, Panipat, Sirsa and Hisar) of Haryana, India to identify the risk factors associated with the usage of CYR revealed that, factors such as cleanliness practices, number of birds kept at the farm and fly density in the farm were significantly associated with the CYR usage in feed for the purpose of fly control. The findings of the present study suggested that 57.69 % farmers had knowledge of CYR and are adding it in the feed of layers as larvicidal agent. Further it was observed that 66.67 % farmers (n=30) use CYR above the recommended levels. Data analysis revealed that factors such as cleanliness practices at the farm (OR: 5.143, 95% CI: 1.403-18.858), number of birds kept at the farm (OR: 17.417, 95% CI: 4.307-75.146) and fly density in the farm (OR: 6.22, 95% CI: 1.836-21.090) were significantly associated with CYR usage in the layer farms.
  • ThesisItemOpen Access
    Molecular detection and characterization of porcine rotavirus from piggery units in Haryana
    (LUVAS,Hisar, 2023) Sheoran,Deepika; Bhanot,Vandna
    Porcine Rotavirus A (PRVA) belong to the family Sedoreoviridae and genus Rotavirus. It is non- enveloped virus and contains double stranded RNA genome of about 16-21 kbp in size with 5’ terminal cap and lacks poly A tail at 3’ end. Genome codes for 6 structural (VP1-4, VP6-8) and 5/6 non-structural proteins (NSP1-NSP5/6). PRVA has been reported from diarrheic and non-diarrheic/asymptomatic pigs from several countries worldwide including India. However, there is no report of molecular characterisation of PRVA from Haryana. So, the present study was designed to detect the presence of PRVA in Haryana by RT-PCR targeting VP6 gene and to characterize it at molecular level. In the present study, out of the total 100 rectal swab samples, 36% (36/100) samples; (38% (19/50) diarrheic and 34% (17/50) non-diarrheic) were tested positive for PRVA by RT-PCR of partial length VP6 gene. Of the positive samples, highest positivity was observed among suckling piglets (45%) followed by weaning piglets (39.13%). None of the samples from growing piglet was found positive for PRVA. Further, the phylogenetic analysis of VP6, VP4 and VP7 genes revealed that the genotype I1, P[13], P[6], G11, G4 and combinations of G4P[6], G4P[13] and G11P[13] are circulating in pig population of Haryana. The most common combination found was G4P[6] (66.66%) followed by G11P[13] (22.22%) and G4P[13] (11.11%). The genotype G11 and the combinations G4P[13] and G11P[13] have been reported for the first time from pigs in India in the present study. The results of this study indicate co-circulation of different genotypes at the same time in pig population of Haryana and reveals high genetic diversity among different PRVA genotypes. These findings are useful for the development of more accurate diagnostic tools and provides information to understand the epidemiology of PRVA in Haryana.
  • ThesisItemOpen Access
    Detection of cyromazine and melamine residues in poultry eggs
    (LUVAS,Hisar, 2023-02) Soni,Deepak; Jadhav, Vijay J.
    Cyromazine (CYR) is an insect growth regulator for fly control at animal farms. It is used as a feed additive in layer farms so that it gets excreted in the droppings of the birds and exert its larvicidal activity in the manure. However, due to this practice, residues of CYR and its metabolite melamine (MEL) also appear in the eggs. A new instrumental method was developed for separation and detection of CYR and MEL on HPLC-UV system. The mobile phase comprised of ACN and 0.05% TFA at pH 2.5 was applied in gradient mode at a flow rate of 1 mL/min. A new method using anhydrous magnesium sulfate, calcium silicate, celite 545 as adsorbents and acidified acetonitrile as extraction solvent was developed for extraction and clean up of CYR and MEL residues in eggs which was found to be quick, easy, cheap, effective, rugged and safe (QuEChERS). The LOD and LOQ of the method was found to be 5.3 and 16.0 μg/kg for CYR and 11.4 and 34.7 μg/kg for MEL, respectively. The accuracy of the method ranged between 84-103 % and precision of the method remained less than 5 % RSD. Linearity studies of spiked sample showed the consistent performance of the method at various concentrations (R²>0.996). A total of 110 egg samples were collected from 52 farms located in 5 different districts of Haryana (Panchkula, Ambala, Panipat, Sirsa and Hisar) and analyzed for the presence of CYR and MEL by applying newly developed QuEChERS method. The mean concentration of the CYR was found to be 42.71 μg/kg with a range of 16.88 -112.61 μg/kg. MEL was not detected in any of the sample analyzed. Estimates of dietary exposure of CYR through consumption of eggs for population in Haryana was found to be 33.40 μg/kg b.w./day which contributed 0.55 % of ADI of CYR. A cross-sectional study was carried out in 52 layer farms situated in 5 districts (Panchkula, Ambala, Panipat, Sirsa and Hisar) of Haryana, India to identify the risk factors associated with the usage of CYR revealed that, factors such as cleanliness practices, number of birds kept at the farm and fly density in the farm were significantly associated with the CYR usage in feed for the purpose of fly control. The findings of the present study suggested that 57.69 % farmers had knowledge of CYR and are adding it in the feed of layers as larvicidal agent. Further it was observed that 66.67 % farmers (n=30) use CYR above the recommended levels. Data analysis revealed that factors such as cleanliness practices at the farm (OR: 5.143, 95% CI: 1.403-18.858), number of birds kept at the farm (OR: 17.417, 95% CI: 4.307-75.146) and fly density in the farm (OR: 6.22, 95% CI: 1.836-21.090) were significantly associated with CYR usage in the layer farms.
  • ThesisItemOpen Access
    The effect of simultaneous lactose hydrolysis and fermentation on the quality of yoghurt
    (LUVAS Hisar, 2023) Hardwari Lal; Sharanagouda B.
    The objective of the present study was to develop Lactose Hydrolyzed Yoghurt by using different concentration of lactase enzyme in yoghurt mix were tried to produce acceptable quality Lactose Hydrolysed Yoghurt with incorporation of stabilizer and emulsifier. Standardization of fat and MSNF in milk were done by Pearson‘s square method. Physico-chemical, rheological, microbiological and sensory characteristics of the products were studied. It was observed that as the level of lactose hydrolysis increased, the penetration value, whey syneresis and acidity of the Lactose Hydrolysed Yoghurt (LHY) increased. Whereas, setting time and sensory attributes were decreased correspondingly with increased degree of lactose hydrolysis. The viscosity of control and all other treatments were decreased with increased temperature at constant shear rate. Based on the sensory evaluation T3 (0.6ml/L lactase enzyme) was adjudged as very much comparable to control. Hence, selected for further storage studies. The developed Lactose Hydrolysed Yoghurt products were stored at 4±1℃ and evaluated at 7 days interval for 28 days. There was decrease in pH and increase in acidity over a period of storage at refrigeration temperature. The standard plate count and starter count increased up to 14th day. But, after that SPC and starter count decreased with the progression of storage period. The yeast and mold count increased with the progression of storage period. Coliforms were absent in all the samples.
  • ThesisItemOpen Access
    Isolation, characterization and in vivo evaluation of therapeutics and prophylactic efficacy of bacteriophage(s) against pathogenic Escherichia coli in chickens
    (2022-10) Jhandai, Punit; Mittal, Dinesh
    The bacterial diseases alone or in combination with other pathogens are causing major economic losses to poultry farmers in the state of Haryana. Avian colibacillosis is one among these diseases which is caused by avian pathogenic Escherichia coli (APEC). The development of resistance to the majority of first-line antibiotics has made the treatment of widely prevalent avian pathogenic E. coli more challenging. So, in this connection, finding alternatives to antibiotics is very much important to control E. coli infections in poultry. Though several alternatives are available to tackle the antimicrobial resiatance problem, but bacteriophages are considered as one of the best remedies. By keeping this in view, the present study was envisaged with the objectives to isolate and characterize pathogenic E. coli and bacteriophage along with in vivo study on therapeutic and prophylactic activity of isolated bacteriophage. Using PCR and Vitek 2 Compact system, a total of E. coli 50 isolates were recovered from tissues of 50 colibacillosis affected flocks. Out of these 45 (90%) isolates were characterized as APEC by multiplex PCR using a set of five virulence genes. The antimicrobial susceptibility testing of E. coli isolates revealed that high antibiotic resistance were observed against imipenem, tetracycline, ciprofloxacin and levofloxacin (96% each). All the 50 isolates were found to be multiple drug resistant (MDR) and the antimicrobial profiling indicated that these isolates could be classified into thirty-eight resistotypes. Moreover, phenotypic characterization of E. coli isolates revealed that 10 (20%) isolates were ESBL producers. On genotypic characterization, 62% isolates were positive for blaTEM gene, whereas, 68% isolates carried intI1 gene. Further, assessment on biofilm formation at 72 h incubation, thirteen (26%) isolates were strong biofilm producers, nine (18%) and twenty-eight (56%) isolates were found to be moderate and weak biofilm producers, respectively. Phylogenetic typing of all the 50 E. coli isolates indicated that phylogroup B1 (38%) was dominant among E. coli isolates followed by A (28%), B2 (10%), unknown/untypable group (8%), F, Clade I/II (6% each) and D (4%). Furthermore, in vivo characterization of multidrug resitant APEC-P02 isolate revealed mean LD50 of 1.12×108 CFU/ml during oral challenge study in day old chicks. Subsequently, a novel lytic phage Escherichia phage PJDM against avian pathogenic E. coli was isolated from poultry farm wastewater and further characterized in vitro and in vivo. The TEM analysis of phage PJDM exhibited an icosahedral head and long non contractile tail, which can be classified as a member of the Siphoviridae family and Caudovirales order. Further, on determination of host range of phage PJDM, the phage exhibited specific lytic activity against several E. coli serotypes (O11, O26, 017, O98, 0120 and O135). Biophysical characterization of phage PJDM indicated that isolated phage remained viable up to 120 days at 30°C, 37°C, 42°C and up to 24 h at pH 5 to 11 with negligible change in its titer. Moreover, one step growth at MOI 0.1 kinetics revealed a latency period of about 20 min and a burst size was 26.5 phages particles/infected cell. Further, the phage PJDM genome has a double-stranded linear DNA molecule and its genomic DNA contains 57,756 base pairs with a GC content of 43.58 % on the basis of whole genome sequencing. Escherichia phage PJDM genome consisted of 98 predicted putative ORFs in genome. Among these 98 genes, 34 genes were predicted to have known functions. The in vitro study on bacteriophage activity against APEC using bacterial challenge test at different MOIs (0.01, 1 and 100) indicated that there were significant decreases in the viability of bacterial strains when infected with an MOI of 100. Then, in vivo evaluation of the efficacy of phage PJDM in day old chicken through oral route at MOI 100 to control E. coli infections revealed that the phage was able to reduce mortality on an average by 20 % and 30% in therapeutic and prophylactic group. The study is first in terms of phage isolation and characterization against colibacillosis causing E. coli from Haryana.