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ThesisItem Open Access Evaluation of haemato-biochemical changes associated with ciprofloxacin therapy in canine pyoderma(Department of Veterinary Biochemistry, College of Veterinary and Animal Sciences, Mannuthy, 2008) Jessy, V; KAU; Jayavardhanan, K KThe present study was undertaken with the objective to assess the haemato-biochemical changes following prolonged ciprofloxacin therapy in dogs affected with pyoderma. The study was conducted in dogs presented to the Veterinary College Hospital, Mannuthy, during the period from August 2007 to March 2008. Among the twenty one dogs studied, twelve completed the full course of antibiotic therapy and post treatment evaluation. The pyoderma confirmed dogs were treated with ciprofloxacin @ 10 mg/kg body weight once daily orally for a period of 14 days. Blood samples collected on day 0, 7 and 14 of treatment and fifth day after the completion of therapy was subjected to haemato-biochemical analysis. Signalment indicated an age wise occurrence of pyoderma and was highest in one to three years of age group (38.10 %). Breed wise occurrence was highest in GSD (38.10 %) and sex wise prevalence was higher in males (52.38 %). Clinical signs and lesions noted in the 21 dogs were combinations of papules and pustules, matting of hairs, erosions, cellulitis, alopecia, scales, hyperpigmentation and pruritus. The antibiotic sensitivity pattern of the gram positive cocci isolates from skin swabs showed maximum sensitivity to ciprofloxacin followed by cefotaxime, cephalexin, gentamicin and amoxycillin. Haematological examination of the blood samples showed no change in TEC, Hb, PCV, ESR and DLC (moncyte and eosinophil) between before, during and after treatment. But a significant variation in TLC, neutrophil and lymphocyte were noticed in dogs with pyoderma. As the treatment progressed, the condition of the animal improved which resulted in decreasing TLC and neutrophil count to normal level. The data obtained on hepatocellular enzymes ALT, AST and other biochemical parameters such as total protein, albumin, A:G and cholesterol suggest that treatment with ciprofloxacin might not have produced any adverse effect on liver tissue. Serum BUN and creatinine levels were found within the normal range. The electrolytes, sodium and potassium were also not altered during the course of treatment. These observations suggest that prolonged administration of ciprofloxacin, at the dose rate mentioned, is not capable of causing any nephrotoxicity. Reduced glutathione also support the above conclusion by eliminating the chance of having any oxidative stress. The present study conclude with the findings that prolonged ciprofloxacin treatment does not produce any deterioration in hepatic and renal system of dogs affected with pyoderma, suggesting ciprofloxacin treatment as a safe and efficacious drug for the complete cure of canine pyoderma.ThesisItem Open Access Effect of piper longum linn.(pippali) in monosodium glutamate toxicity in rats(Department of Veterinary Biochemistry, College of Veterinary and Animal Sciences, Mannuthy, 2008) Mariyamma, Thomas; KAU; Sisilamma, GeorgeThe present study was conducted to evaluate the effect of ethanolic extract of fruits of Piper longum in monosodium glutamate toxicity in rats. Treatment as well as protective effects of the plant extract against MSG toxicity were studied. The experiments were carried out in adult male Wistar rats, which were divided into seven groups. The treatment study was conducted in four groups viz; G0- normal control, G1-positive control, G2 and G3- two treatment groups. G1, G2 and G3 were administered with MSG at a dose rate of 8 mg/g body weight p.o. for 20 days followed by treatment of G2 and G3 with Piper longum extract at dose rates of 300 mg/kg and 600 mg/kg b.w p.o. respectively for 14 days.. Blood collection and weight recording of these animals were carried out on days 0, 21, 28 and 35 of experiment and then the animals were euthanized. The remaining three groups viz; G4, G5 and G6 were subjected to protective study where G4 served as normal control, G5, positive control and G6 was administered with Piper longum extract at 300 mg/kg dose rate along with MSG for 20 days. Blood samples were collected and the animals were weighed on days 0 and 21 of experiment followed by euthanasia. The oxidative stress and subsequent damage to liver and kidney were assessed by measuring the biochemical parameters viz; activities of serum ALT, AST, concentration of serum triacylglycerol, total cholesterol, bilirubin, total protein, albumin, A:G ratio, urea, creatinine and the levels of serum and tissue (liver and kidney) lipid peroxides and GSH. From the euthanized animals liver and spleen were separated and weighed. Representative samples of liver and kidney tissues were subjected to histopathological examination. Administration of MSG induced a significant increase in the body weight, weight of liver and spleen. The increase in body weight was gradual and became significant only on day 35. Oxidative injury to the tissues of liver and kidney was evident from the increased level of lipid peroxides, decreased level of GSH and increased activities of serum ALT and AST. There was also an increase in the levels of serum triacylglycerol, cholesterol and urea. Histopathological examination of the liver and kidney of positive control animals revealed necrosis of hepatocytes in the para cortical and midzonal areas of liver and diffuse cortical tubular degeneration, occasional necrosis and shrinkage of glomeruli of the kidney. However, the hepatic and nephro toxicities caused by MSG were not so severe to alter the levels of total protein, albumin, A:G ratio, bilirubin and creatinine in serum. Piper longum extract at both the dose levels proved to be effective in treating the toxicity induced by MSG and helped to bring back the body weight and weight of spleen near to that of the control, significantly reduced the lipid peroxides and increased the GSH levels in serum, liver and kidney. Treated groups also showed a significant reduction in serum ALT and AST activity, ameliorated the MSG induced hyperlipidemia and normalized the histological architecture of both the liver and kidney. Among the two dose rates, only the 300 mg/kg dose rate was effective in maintaining the liver weight near to that of the control and this dose rate was found to be more effective in alleviating the toxicity caused by MSG. Co-administration of Piper longum extract and MSG prevented the abnormal increase in the weight of vital organs, level of lipid peroxides in serum, liver and kidney, while no increase was observed in the level of GSH. Significant decrease was also observed in the levels of serum AST, triacylglycrol and total cholesterol, whereas no change was observed in the level of ALT and urea. Although, ethanolic extract of Piper longum fruits at 300 mg/kg dose rate could offer significant protection against the induction of toxicity by MSG, it appears that the dose rate is insufficient to provide a complete protection against the oxidative injury.ThesisItem Open Access Clinico-therapeutic studies on canine microfilariasis(Department of Clinical Veterinary Medicine, College of Veterinary and Animal Sciences, Mannuthy, 2009) Ambily, V R; KAU; Usha, Narayanapillaietc. A study on canine microfilariosis was conducted in the Department of Clinical Veterinary Medicine, College of Veterinary and Animal Sciences, Mannuthy during the period of 2007-2009. Hundred dogs of both sexes belonging to various breeds and above 6 months of age presented to Veterinary College Hospital, Mannuthy and University Veterinary Hospital, Kokkala from different parts of Kerala with clinical signs suggestive of microfilariosis were screened for microfilaria by wet film examination. Wet film examination revealed that 80% of dogs were positive for microfilaria. Staining of blood smear with giemsa (1:10) demonstrated that 16 out of 80 dogs were positive for sheathed microfilaria and remaining were nonsheathed. Out of these 50 (nonsheathed) microfilaremic dogs were selected and treated at random with five schedules of treatment so that each schedule consisted of ten animals each (Group I, II, III, IV and V). Sheathed microfilaraemic animals were considered as a separate group for treatment trial. All these animals were subjected to periodic wet film examination on 2nd, 4th and 7th day of treatment to assess the treatment response. High infestation rates were recorded in male dogs of 2-4 years of age than females irrespective of the type of microfilariae. High incidence of microfilariosis with non sheathed and sheathed microfilariae was observed in GSD and Labrador breeds respectively. Diagnosis was made by parasitological studies, immunological and molecular techniques, clinical investigations and haematobiochemical analysis. In wet film examination distinct patterns of motility was observed with the type of microfilariae present. The speciation of microfilariae were done based on morphological characteristics in giemsa stained smears, acid phosphatase enzyme activity, immunospot test and PCR analysis and sequencing of amplicon. The different species of microfilariae identified were that of Dirofilaria repens, Dipetalonema reconditum, Brugia malayi and Brugia pahangi. Of which Brugia malayi and Brugia pahangi were sheathed. Results of micrometry, staining, immunospot test and molecular studies revealed that the newly identified parasite were similar to that of Brugia malayi in human beings. This is the first report of detection of Brugia malayi in dogs for which no previous reports were available in pubmed or other literature data bases. Infact this is also the first report of Brugia pahangi in a dog from India and Dipetalonema reconditum from dogs of Kerala. Detailed clinical investigations included ECG, ultrasonography and radiography were conducted to visualize the abnormalities encountered with vital organs. Haematobiochemical studies of dogs affected with both non sheathed and sheathed microfilariae revealed mild anaemia with severe leucocytosis, neutropenia, lymphocytosis, eosinophilia, elevated ESR and severe thrombocytopaenia, hyperproteinemia with hyperglobulinaemia and non significant reduction in AG ratio, increased serum ALT, AST, ALP , BUN and creatinine values could be observed when compared to healthy controls. Qualitative urinalysis revealed proteinuria with reduced specific gravity. Quantitative analysis of urinary markers revealed elevation of NAG, UPC, γGT and ALP in microfilaraemic dogs. The elevated levels of serum total protein, globulin, serum enzymes like ALT and ALP and nonsignificant reduction in AG ratio suggestive of liver pathology in microfilaraemic dogs. Elevated levels of BUN, creatinine, urine protein creatinine ratio, NAG, ALP, proteinuria with low specific gravity confirmed the renal involvement in microfilaraemic dogs irrespective of the type of microfilaria involved in the disease process. This multiorgan pathology in canine microfilariosis suggested the involvement of toxic and immunological effects of these parasite in the pathogenesis of the disease. The treatment response was evaluated by the periodic clearance of microfilariae on wet blood film examination, remission of clinical signs and the improvement in haematobiochemical alterations. Single oral dose of ivermectin @ 100 µg/kg body weight can be selected as a treatment modality for microfilariosis due to Dirofilaria repens and Dipetalonema reconditum in dogs. Levamisole hydrochloride @ 10 mg/kg body weight for seven days was the only effective treatment for microfilariosis due to Brugia malayi in dogs. Result of post treatment values of hepatic and renal function test revealed that many of the parameters like ALT, ALP and BUN were still in elevated level. Two animals died during the course of treatment were subjected to post mortem examination. The gross and hispathological examination revealed lesions in heart, lungs, liver and kidneys in microfilaraemic dogs. Myofibrillar fragmentation, atlectasis, thromboemboli formation, portal hepatitis and chronic interstitial nephritis were the major lesions observed. Based on above studies it concluded that follow-up evaluation of these parameters could be a relevant approach to find out the therapeutic effectiveness. A therapeutic plan should consists of both specific and clinically supportive treatments to improve hepatic and renal function. A study on canine microfilariosis was conducted in the Department of Clinical Veterinary Medicine, College of Veterinary and Animal Sciences, Mannuthy during the period of 2007-2009. Hundred dogs of both sexes belonging to various breeds and above 6 months of age presented to Veterinary College Hospital, Mannuthy and University Veterinary Hospital, Kokkala from different parts of Kerala with clinical signs suggestive of microfilariosis were screened for microfilaria by wet film examination. Wet film examination revealed that 80% of dogs were positive for microfilaria. Staining of blood smear with giemsa (1:10) demonstrated that 16 out of 80 dogs were positive for sheathed microfilaria and remaining were nonsheathed. Out of these 50 (nonsheathed) microfilaremic dogs were selected and treated at random with five schedules of treatment so that each schedule consisted of ten animals each (Group I, II, III, IV and V). Sheathed microfilaraemic animals were considered as a separate group for treatment trial. All these animals were subjected to periodic wet film examination on 2nd, 4th and 7th day of treatment to assess the treatment response. High infestation rates were recorded in male dogs of 2-4 years of age than females irrespective of the type of microfilariae. High incidence of microfilariosis with non sheathed and sheathed microfilariae was observed in GSD and Labrador breeds respectively. Diagnosis was made by parasitological studies, immunological and molecular techniques, clinical investigations and haematobiochemical analysis. In wet film examination distinct patterns of motility was observed with the type of microfilariae present. The speciation of microfilariae were done based on morphological characteristics in giemsa stained smears, acid phosphatase enzyme activity, immunospot test and PCR analysis and sequencing of amplicon. The different species of microfilariae identified were that of Dirofilaria repens, Dipetalonema reconditum, Brugia malayi and Brugia pahangi. Of which Brugia malayi and Brugia pahangi were sheathed. Results of micrometry, staining, immunospot test and molecular studies revealed that the newly identified parasite were similar to that of Brugia malayi in human beings. This is the first report of detection of Brugia malayi in dogs for which no previous reports were available in pubmed or other literature data bases. Infact this is also the first report of Brugia pahangi in a dog from India and Dipetalonema reconditum from dogs of Kerala. Detailed clinical investigations included ECG, ultrasonography and radiography were conducted to visualize the abnormalities encountered with vital organs. Haematobiochemical studies of dogs affected with both non sheathed and sheathed microfilariae revealed mild anaemia with severe leucocytosis, neutropenia, lymphocytosis, eosinophilia, elevated ESR and severe thrombocytopaenia, hyperproteinemia with hyperglobulinaemia and non significant reduction in AG ratio, increased serum ALT, AST, ALP , BUN and creatinine values could be observed when compared to healthy controls. Qualitative urinalysis revealed proteinuria with reduced specific gravity. Quantitative analysis of urinary markers revealed elevation of NAG, UPC, γGT and ALP in microfilaraemic dogs. The elevated levels of serum total protein, globulin, serum enzymes like ALT and ALP and nonsignificant reduction in AG ratio suggestive of liver pathology in microfilaraemic dogs. Elevated levels of BUN, creatinine, urine protein creatinine ratio, NAG, ALP, proteinuria with low specific gravity confirmed the renal involvement in microfilaraemic dogs irrespective of the type of microfilaria involved in the disease process. This multiorgan pathology in canine microfilariosis suggested the involvement of toxic and immunological effects of these parasite in the pathogenesis of the disease. The treatment response was evaluated by the periodic clearance of microfilariae on wet blood film examination, remission of clinical signs and the improvement in haematobiochemical alterations. Single oral dose of ivermectin @ 100 µg/kg body weight can be selected as a treatment modality for microfilariosis due to Dirofilaria repens and Dipetalonema reconditum in dogs. Levamisole hydrochloride @ 10 mg/kg body weight for seven days was the only effective treatment for microfilariosis due to Brugia malayi in dogs. Result of post treatment values of hepatic and renal function test revealed that many of the parameters like ALT, ALP and BUN were still in elevated level. Two animals died during the course of treatment were subjected to post mortem examination. The gross and hispathological examination revealed lesions in heart, lungs, liver and kidneys in microfilaraemic dogs. Myofibrillar fragmentation, atlectasis, thromboemboli formation, portal hepatitis and chronic interstitial nephritis were the major lesions observed. Based on above studies it concluded that follow-up evaluation of these parameters could be a relevant approach to find out the therapeutic effectiveness. A therapeutic plan should consists of both specific and clinically supportive treatments to improve hepatic and renal function. Further studies are warranted to elucidate the possible role of dogs in the transmission of human filariasis and to develop a suitable therapeutic approach to treat canine microfilariosis in the increasing ewe of chronic renal diseases in dogs.ThesisItem Open Access Evaluation of intranasal canine parvovirus vaccination in dogs(Department of Veterinary Epidemiology and Preventive Medicine,College of Veterinary and Animal Sciences, Mannuthy, 2010) Murugesan, V; KAU; Tresamol, P VPresent study was carried out to evaluate the potency of intranasal canine parvovirus vaccination in dogs at the Department of Veterinary Epidemiology and Preventive Medicine, College of Veterinary and Animal Sciences, Mannuthy during the period 2009-2010. The post vaccination immune response of intranasal CPV vaccination and the interference of MDA were assessed by HI and SN tests. The vaccine performance was studied in twelve unvaccinated healthy pups of six to eight weeks age. The blood samples were collected before vaccination for assessment of the CPV antibodies by HI test and the pups were grouped into two viz., Group 1 (G1) having HI titre of ≤80 (range, 0 to 80) and Group 2 (G2) having HI titre of >80 (range, 160 to 320). Intranasal vaccination has been done based on two different schedules viz., Schedule I and II. Schedule I was performed by using one drop of attenuated CPV-2 antigen each dose containing 103 TCID50 antigenic mass through intranasal route on day zero and day two. Schedule II was performed using the same antigen intranasally on day zero, day two and day four. Seroconversion was considered when pups developed a fourfold or greater increase in HI titre to a level ≥ 80. Hundred percent of the pups were seroconverted in both groups and had protective titre against CPV disease. In group I animals, the range of geometric mean HI titre from day zero to day 90 post vaccination was 30.58 to 2560. The titre also increased from day seven to day 90 and the differences between consecutive sampling intervals were statistically significant (P<0.05). In group II animals, the range of geometric mean HI titre recorded from zero to 90th day post vaccination were 201.58 to 1436.75. Active immune responses after vaccination were observed in all pups but a fall in immune response was noticed on 7th day with a geometric mean HI titre of 126.99 compared to the geometric mean HI titre of 201.58 on day zero. Statistically significant difference in titre was observed only on 90th day. The pattern of seroconversion in SN titres on day zero to day 90 post inoculation was similar to that of HI test in both groups. All the pups belonging to group I had seroconversion within a week or two after vaccination. All pups belonging to group II had a good titre of seroconversion on day 90, and surpassed the maternal antibody interference with a geometric mean HI titre as high as 201.58. In both groups, slight fall in seroconversion titre was noticed on 60th day post vaccination in all pups which may be due to some stress or immunogenic unresponsiveness. None of the vaccinated pups developed anaphylaxis or other adverse reactions attributable to the vaccine and no viral shedding in the faeces following vaccination. All the pups in both tested groups survived in good health during the post vaccination monitoring period of one year and none of the pups showed any clinical signs of CPV disease. The vaccine was well tolerated by the dogs and there were no adverse effects. The findings of this study suggested that good protection against CPV infection may be achieved by the use of attenuated CPV-2 antigen with a titre of 103TCID50 per dose administered intranasally based on both schedules. This minimal vaccine mass was able to induce active immune responses in all pups with MDA titres of ≤ 80 with a range of zero to 80 according to schedule I and > 80 with a range of 160 to 320 according to schedule II by intranasal route and may confer more adequate protection than that of oral and parentral routes. This method of immunization was proved simple, convenient, reliable, safe, painless and efficacious. Hence, both vaccination schedules could be recommended for immunoprophylaxis under field conditions against CPV disease in dogs.