PG Thesis

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    Effect of a composite mixture of Emblica officinalis, Terminalia chebula and Terminalia bellirica on aflatoxicosis in rabbits
    (Centre for Excellence in Pathology, College of Veterinary and Animal Sciences, Mannuthy, 2009) Indu, K; Lalithakunjamma, C R
    Among the various mycotoxins, aflatoxins have been the subject of most intensive research because of the extremely potent cytotoxic and carcinogenic effects. Mycotoxicosis and its counteraction have received greater attention by researchers in the last few decades. In this context an evaluation study on the protective effect of a herbal composite mixture triphala (containing Emblica officinalis, Terminalia chebula and Terminalia bellirica) on aflatoxicosis in rabbits was carried out. The study was conducted for a period of two months with twenty four rabbits divided into three groups of eight each. Rice culture containing 27 ppm of aflatoxin was added appropriately to the ration to get a final concentration of 0.5 ppm. Group I was given 0.5 ppm aflatoxin contaminated feed and group II was given combination of triphala (four per cent) and aflatoxin (0.5 ppm ) contaminated diet. Group III was provided with control diet. There were reduction in body weight gain, Hb, PCV, TLC, lymphocyte count while heterophil count, AST, ALT and ALP levels showed an increase in the toxin fed animals. Addition of triphala at four percent level in the aflatoxin contaminated feed effectively counteracted these changes. The gross and histopathological changes due to aflatoxin were reduced in triphala group. There was widespread vascular changes, extensive necrotic changes, bile duct proliferation, biliary hyperplasia and cholangiocellular carcinoma in the liver and necrotic changes in the kidney of toxin group. These changes were reduced in intensity in triphala group and there was no development of tumors in liver. Regenerative process were well pronounced in the liver and kidney of triphala group. Hence the present study revealed that supplementation of composite mixture of Emblica officinalis, Terminalia chebula and Terminalia bellirica at four per cent level could counteract the toxic effects of aflatoxicosis in rabbits.
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    Production and evaluation of vaccines employing Pasteurella multocida A:1 grown under different growth conditions
    (Department of Microbiology, College of Veterinary and Animal Sciences, Mannuthy, 2007) Raja Gopal, R; Krishnan Nair, G
    A research work was undertaken to prepare effective vaccines against P. multocida grown under different conditions and their immunopotency assessed in one month old ducklings. The purity of the Pasteurella multocida A: 1 strain (DP1) was confirmed as per standard procedures. Pathogenicity of the isolate was assessed in six to eight weeks old mice. The isolate killed the intraperitoneally inoculated mice within eight hours and within 24 h when injected by subcutaneous route. The organism was grown in different media to assess the amount of capsular material produced. The media employed were DSA, DSA supplemented with 10 per cent FBS and DSA supplemented with 10 per cent FBS and 0.5 per cent yeast extract. The capsule enhancement was measured by capsule demonstration using Maneval staining and by characterization of crude capsular extract. The capsules of the organisms grown in capsule enhancement media when demonstrated using Maneval staining were discernibly larger and denser, when compared to the organisms grown in DSA alone. The capsular polysaccharides increased by approximately 1.6 times when the organism was grown in capsule enhancement media containing 10 per cent FBS and by about 2.06 times when grown in media supplemented with FBS and yeast extract. The potential of the organism to form in vitro biofilm was assessed by growing the organism in nutrient restricted conditions. The organism was grown in 0.32 per cent TSB supplemented with an inert substrate called bentonite clay, for the bacteria to attach and colonize. For quantification of biofilm, plate count by spread plate method was employed and the results showed that the biofilm cells reached a peak on the third day of incubation with an average count of 1.54 ×106 CFU/g of bentonite clay while the planktonic cells were found to be maximum on day one post inoculation, with a peak count averaging about 8.10 ×108 CFU/ml of the broth. Maneval staining of late logarithmic phase of three day old biofilm culture revealed heavily capsulated cells of P. multocida attached as large aggregates and appearing as chains of coccobacillary cells. Also they appeared as a meshwork of aggregated and chain forming cells. The capsular polysaccharide estimation of biofilm cells revealed a 3.25 times increase over the planktonic bacteria grown in DSA alone. The biofilm cells on nutrient agar after 24 h at 37°C produced some colony morphotypes characterized by radiating strands from centre to periphery and wavy margin. Median lethal dose (LD50) of P. multocida when determined in one month old ducklings was 23 cells. In the present study, it was unable to arrive at the median lethal dose in six month old ducks as only one duck died after 48 h of virulent challenge. Oil adjuvant formalin inactivated bacterin vaccines were prepared from DP1 grown in TSB, capsule enhancement medium and under biofilm mode and performed the sterility, safety and potency tests of the vaccine employing standard procedures. A total of 160 four week old ducklings were divided into four groups with 40 birds in each group and the first three groups were vaccinated with ordinary bacterin, capsule enhanced bacterin and biofilm vaccine respectively. The fourth group served as control. The birds were vaccinated with 0.5 millilitre of vaccine intramuscularly in the thigh region. Blood was collected from all the ducks pre-vaccination, at weekly intervals upto 28th day post PV and on day 42 PV by cardiac puncture or by jugular venipuncture. Passive haemagglutination using GA-SRBC sensitized with sonicated antigen of DP1 was used to measure the humoral immune response. The IHA titres obtained for biofilm vaccine group on day 14 was very much higher than the other two groups. The antibody titre was observed from day seven onwards for all the groups. All the vaccine groups have shown significant difference from the control group at all the stages of the study. Groups I and II were having no significant difference in their mean titres during the entire study. On homologous challenging, biofilm vaccine gave higher protection rates of 70 and 90 per cent than the 60 and 80 per cent protection rates of ordinary and capsule enhanced bacterins, when challenged with 200 and 100 LD50 doses respectively. Biofilm vaccine was proved to be the best among the three vaccines tried. The capsule enhanced vaccine did not provide any additional advantage over the ordinary bacterin vaccine. Elaborate field trials are to be done before advocating the vaccine for commercial use.
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    Investigation on sudden death in pigs
    (Department of Clinical Medicine, College of Veterinary and Animal Sciences, Mannuthy, 2006) Manjusha, A; Baby, P G
    Study of “Investigation on sudden death in pigs’’ was conducted in pigs from various pig farms in Kerala, where cases of sudden death were reported. In these farms around 12.4 per cent of the total pigs were lost due to sudden death. Post mortem examination were carried out in six animals. Clinical materials were collected from the forty-five contact animals and six apparently healthy control animals. Occurrences of sudden death were common in three to six month old piglets. Most of the animals died without any premonitory clinical signs. All these animals were fed with uncooked swill mainly containing chicken waste. Some of the contact animals showed clinical signs like elevated temperature, congested mucous membranes and diarrhoea. On cultural examination of the samples from the two farms growth of the gram-negative organisms were obtained. Transmission of these organisms to these pigs may be from the uncooked swill fed to them. On antibiotic sensitivity testing cefotaxime and ceftriaxone were found to be effective against these organisms. Leukocytoses with neutrophilia were observed in animals of group III where as normal leukogram observed in animals of group II. Serum biochemical studies revealed an elevated level of creatine kinase, lactate dehydrogenase and aspartate aminotransaminase in animals without any obvious clinical sign of illness, indicating involvement of heart in the pathological condition. Animals with obvious clinical signs of illness revealed an elevated level of lactate dehydragenase, indicating damage of the tissues. Gross pathology on post mortem revealed lesions in heart, liver, brain, lymph nodes and kidney. Haemorrhages in the heart were the major findings. Prominent histopathological changes were noticed in heart and brain. Heart showed varying degrees of inflammation, degeneration and necrosis. Stomach, intestine, lung, liver and kidney also showed pathological changes. For all the animals under study supplementation of mineral mixture containing vitamin E and Selenium were advised. Animals with clinical signs of illness were treated with cefotaxime. It prevented further mortality in these farms.
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    Antipyretic and CNS activity of seeds from red and white types of lotus (nelumbo nucifera) in albino rats
    (Department of Pharmacology and Toxicology, College of Veterinary and Animal Sciences, Mannuthy, 2006) Deepa, P K; Usha, P T A
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    Anti-inflamatory and analgestic actions of red and white lotus seeds (nelumbo nucifera) in albino rats
    (Department of Pharmacology and Toxicology, College of Veterinary and Animal Sciences, Mannuthy, 2006) Vikrama Chakravarthi, P; Gopakumar, N
    Anti-inflammatory and analgesic effects of Nelumbo nucifera (lotus) seeds, red and white types were assessed and compared in albino rats. The carrageenin induced paw oedema was one of the acute inflammatory models followed in anti-inflammatory screening. The acute and chronic type pain models viz., tail flick method and experimental neuropathy model of analgesic activity, respectively, were adopted for analgesic screening. Diclofenac potassium at the rate of 3 mg/kg was used as a standard drug for both the studies. In cyclooxygenase-2 enzyme inhibition assays the celecoxib @ 10mg/kg was used as a standard drug. Both red and white types of lotus seeds at the dose rate of 400 mg/kg and 600 mg/kg were taken for the anti-inflammatory and analgesic studies. Anti-inflammatory effect of red and white lotus seeds was found to be effective in all phases of carrageenin induced inflammation. The higher dose groups of lotus seed extracts were revealed more inhibition than their corresponding lower dose. While comparing all groups, the higher dose group of white lotus seed, exhibited more pronounced inhibition of paw oedema than others. Analgesic effect was found to be significant in both acute and chronic analgesic models. The analgesic activity was more in tail flick method when compared to experimental neuropathy model. The cyclooxygenase-2 (COX-2) enzyme inhibition assays in both ELISA and spectrophotometer showed significant effect than control. The percentage of inhibition of COX-2 was more evidenced in both lotus seed extract of higher dose groups than its lower dose. However, the higher dose groups of white lotus seed exhibited more control over inhibition of COX-2 enzyme than others. . In case of biochemical parameters in both anti-inflammatory and analgesic screening the serum cholesterol level was found to be decreased in treatment groups when compared to control. Even though, there was a reduction in serum cholesterol level, all the values were within the normal range. The rise in serum level of AST and ALT in both inflammation and analgesic models were noticed in lower dose treated group. Conversely, there was a reduction in level of enzyme was noticed in higher dose treated groups. However, the values were with in the normal level. Both the studies showed the haematological parameters viz., total leukocyte and differential counts were within the normal range in all groups, even though a non-significant increase in neutrophil count than lymphocyte was noticed in carrageenin induced inflammation and tail flick method in rats.
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    Metabolic and endocrine profile of crossbred pre-ruminant calves under extended colostrum feeding
    (Department of Physiology, College of Veterinary and Animal Sciences, Mannuthy, 2003) Babitha, V; Philomina, P T
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    Effect of human chorionic gonadotrophin,gonodotrophin releasing hormone and prostaglandin F2 alpha in augmenting conception rate of repeat breeder cattle
    (Department of Animal Reproduction, College of Veterinary and Animal Sciences, Mannuthy, 2003) Velayudhakumar, R; Sreekumaran, T
    Repeat breeding in cattle especially due to ovulatory disturbances has become a frustrating problem under field conditions. The present study was aimed to formulate an effective and economic therapy for this problem. Hence three therapeutic approaches viz. human chorionic gonadotrophin, Gonadotrophin releasing hormone and prostaglandin F2 alpha were undertaken, and its efficacy and economic viability was compared. The material for the study consisted of repeat breeder cross-bred cows and heifers presented at Artificial insemination centre and bull station, attached to the Department of Animal Reproduction, College of Veterinary and Animal Sciences, Mannuthy, Thrissur. Occurrence of ovulatory disturbance were studied and white side test was done to exclude those with infection Overall occurrence of anovulation and delayed ovulation were 7.38 and 15.77 per cent.Anovualtion in cows and heifers were 5.21 and 17.82 and delayed ovulation was 14.7 and 8.8 per cent respectively. On study of . characteristics of cervical mucus, clear colour (93 per cent), thick consistency (43.47 per cent) and atypical crystallization (57.97 per cent) were found to be high in repeaters with ovulatory disturbances. Spinnbarkeit value varied significantly (t' = - 4.2779, P < 0.01) where as pH variation was non significant (t' = 2.37, P > 0.05) between normal and repeaters. Changes in the tubular tract such as vulval oedema, hyperaem ia of vaginal mucosa and uterine tonicity were found to be medium and low in most of the repeat breeders. Repeat breeders with ovulatory disturbances were randomly allotted to three groups of twelve each group-A, group B and group C. Intramuscular administration of Human chorionic gonadotrophin (Chorulon, 1500.I.U),GnRH analogue (Buserelin acetate -Receptal, l Ougjand Prostaglandin F2 alpha analogue (Luprostiol - Prosolvin, 3 mg) were done immediately after insemination in group-A ,group B and group C respectively. There was a significant reduction in duration of oestrum in group A (50.17 to 38.08 hours) and group B (50.08 to 36.17 hours) and non significant in group C. Ovulatory response was 33.33, 75.00 and 66.66 per cent respectively in group A, Band C. Conception rate was 25,50,50 and 22.22 per cent respectively in group A, B, C and repeaters given double insemination at 24 hours interval. The cost for treatment per animal in rupees and amount required per conception in rupees was 220, 103,44, and 35 and 880, 206, 88 and 157.5 respectively in group A, B, C and those given double insemination. Hence it is inferred that PGFz alpha treatment immediately after insemination is preferred to treatment with GnRH or hCG or double insemination at 24 hour interval and that administration ofGnRH is found better than hCG in repeat breeding due to ovulatory disturbances in cattle.
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    Clinico-therapeutic studies on ehrlichiosis in dogs
    (Department of Veterinary Epidemiology and Preventive Medicine, College of Veterinary and Animal Sciences, Mannuthy, 2003) Smitha, J P
    The present study was undertaken to get a detailed understanding on clinico-therapeutic, haematological and biochemical aspects of ehrlichiosis in dogs. A total of 64 animals showing symptoms suggestive of the disease were taken as the study group. Diagnosis was based on examination of blood/buffy coat smear and by IF A test as well. Examination of blood/buffy coat smear revealed Ehrlichia canis morulae in the cytoplasm of leukocytes in 16 animals. IF A test, detected antibodies in 42 animals. 0 age, sex or breed predilection for the disease was noticed. Most frequent clinical signs observed were fever, anorexia/selective appetite, lymphadenomegaly, depression and congested mucous membrane. Haematological abnormalities mainly encountered included normocytic normochromic anaemra, thrombocytopenia and monocytosis. Serum biochemical abnormalities observed were elevated levels of serum ALT, AP, total protein, globulin, creatinine and low levels for albumin and lower values for A/G ratio. A therapeutic trial with doxycycline and prednisolone gave encouraging clinical response and good uneventful recovery. The results of the present study confirm that ehrlichiosis is a disease that is prevalent in our area and treatment with doxycycline can be considered as the therapy of choice for ehrlichiosis in dogs.
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    Prevalance and pathology of brain disorders in cattle
    (Centre of Excellence in Pathology, College of Veterinary and Animal Sciences, Mannuthy, 2003) Balasubramanian, M; Divakaran Nair, N
    The present study was undertaken to assess the prevalence and pathology of various brain disorders in cattle. The data collected from the autopsy records maintained at the Centre of Excellence in Pathology, Mannuthy and fifty-two samples of brain obtained from both the slaughtered cattle and autopsy cases formed the basis of the, study. Gross lesions were not so evident except for the meningeal congestion, which accounted to 70 per cent of the cases. Congestion and haemorrhages were found consistently in the slaughtered cattle, which could be attributed to stunning effect. The other major lesions were well appreciated by detailed histopathological examination of ten different areas of the brain in each of the cases. These included vascular changes, neuronal degeneration, inflammatory changes, gliosis, pigmentation and neoplasia. Two cases revealed intra neuronal cytoplasmic inclusions of which one was due to rabies and the other was attributedto viral etiology. Bovine spongiform encephalopathy was not found to be present in any of the cases in the present study. Based on the observations made here, it was suggested that diagnosis of rabies by impression smear examination alone is not sufficient and needs examination by FAT or Immunoperoxidase techniques. The incidence of pathological disorders encountered was relatively high. Thus, the need and scope for investigation into the pathological disorders of bovine brain were highlighted.
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    Evaluation of the immunomodulatory effect of Emblica officinalis (amla) fruit pulp extracts in mice
    (Department of Pharmacology and Toxicology, College of Veterinary and Animal Sciences, Mannuthy, 2003) Suja Rani, S; Chandrasekharan Nair, A M
    The immunomodulatory activity of alcoholic and aqueous extracts of Emblica offictnalis was investigated on the basis of their effects on humoral, cell mediated and cellular immune mechanism in mice. The extracts were also qualitatively tested for the presence of various active principles in it. One hundred and forty four mice taken for the study were divided into two in which one group was tested with alcoholic extract while the other with aqueous extract. The extracts were fed at two dose levels ie, l00mg and 200 mg/kg bodyweight for 19 days. The controls in both groups received vehicle alone (five percent gum acacia). Various physiological, biochemical, haematological and immunological parameters like bodyweight, organ weight, total leukocyte count, differential leukocyte count, serum total protein, serum globulin, Haemagglutination (HA) titre, Delayed Type of Hypersensitivity (DTH), Macrophage Migration Index (MMI) and NitroBlue Tetrazolium (NBT) dye reduction test were performed for evaluating the immunomodulatory potential of the extract. Both the extracts were found to increase the bodyweight and spleen weight significantly when fed at a higher dose rate of 200mglkg for 19 days. The total leukocyte count was increased to a maximum of 13.9O±2.05 and 12.77±<>.78 xI03/cu.mm respectively in aqueous and alcoholic extract treated groups on 1~ day of experiment, compared to control groups (7.l8±<>.72 and 7.IS± 0.72). Lymphocytic leukocytosis was seen after drug treatment. Serum total protein and globulin levels were also increased by the administration of extracts of Emblica. The drug administration increased the globulin concentration to 1.88±<>.42 and 1.97±0.21goA» on 19th day for alcoholic and aqueous extract treated groups respectively which was significantly higher than the control groups. The increase in HA antibody titre indicated the augmentation of humoral immune response to SRBC by Emblica officina/is. Administration of Amla extracts significantly increased cell mediated immune response as evidenced by increase in DTH response. Both the extracts were found to increase the macrophage migration area to 10.93±2.21 and 11.87±3.54 mm' in aqueous and alcoholic extract treated group respectively on 19th day of experiment compared to the controls (5.33±1.61 and 4.78±2.18). Thus a 1.5 to 2.5 fold increase in MMI could be noticed. The results of NBT test gave a maximum of 53.28 percent increase in respiratory burst activity of macrophage in aqueous extract treated group while the alcoholic extract treated group had a maximum of 46.04 percent increase from their controls. The phytochemical study of the Emblica extracts revealed the presence of active principles like tannins, flavonoids, glycosides, phenols, diterpenes, triterpenes and saponins in it. Thus the present study establishes the positive immunomodulatory activity of dried Emblica officinalis fruit pulp extracts, in a concentration dependent manner acting via humoral, cell mediated and cellular immune response.