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ThesisItem Open Access EARLY DIAGNOSIS OF PSEUDOPREGNANCY IN GOATS BY ULTRASONOGRAPHY AND THERAPEUTIC MANAGEMENT USING CLOPROSTENOL SODIUM(COLLEGE OF VETERINARY AND ANIMAL SCIENCES, POOKODE, WAYANAD, KERALA VETERINARY AND ANIMAL SCIENCES UNIVERSITY, 2023-04-11) NEEMA CHERIAN; Dr. Abdul Azeez C. P.The research was undertaken in goats with the objective of early diagnosis of pseudopregnancy using ultrasonography, estimation of serum progesterone concentration, treatment with three doses of cloprostenol sodium and evaluation of conception rate post treatment. The pseudopregnant does (Group I, n=8) diagnosed on day 60 post-breeding using B-mode ultrasonography (anechoic fluid filled uterus with hyperechoic trabeculae) were treated with three doses of 125µg of cloprostenol sodium (i/m) on the day of diagnosis (day 0), day 10 and day 20. The response to treatment were evaluated and compared with normal cycling does (Group II, n=8). Serum progesterone concentration was estimated on day 0, day of evacuation of uterine fluid, on days 10 and 20, day of induced oestrus, on days 21 and 60 post breeding in group I and on day of oestrus, on days 10, 21 and 60 post breeding in group II. Out of 254 goats screened for pregnancy, 15 does were identified as pseudopregnant (5.90%). Among various risk factors related to pseudopregnancy, significant relationship was identified with age (P=0.022), parity (P=0.047) and season (P=0.001). Ultrasonographic examination on different days revealed uterine wall thickness in pseudopregnant does reduced significantly (p <0.05) from day 0 (0.63 ±0.024 cm) to day 20 (0.58 ±0.025). Serum progesterone concentration (RIA) on day 0 and day 60 post breeding varied significantly (P < 0.01) between group I (2.73 ± 0.19 ng/mL) and group II (3.91± 0.19 ng/mL), respectively. The treatment protocol followed was sufficient to induce oestrus in 100 per cent pseudopregnant does. Conception rate varied non-significantly (P >0.05) between group I (50%) and II (75%). Therefore, it is recommended that pseudopregnancy can be diagnosed as early as 60 days post breeding and can be effectively treated using three doses of cloprostenol sodium 10 days apart.ThesisItem Open Access SURGICAL MANAGEMENT OF TRAUMATIC SPINAL INJURIES IN SNAKES USING MODIFIED MICROPLATE SPINAL FIXATION TECHNIQUE(CENTRE FOR WILDLIFE STUDIES, POOKODE, WAYANADKERALA VETERINARY AND ANIMAL SCIENCES UNIVERSITY, 2023-04-10) NITHIN K.; Dr. George ChandyModified microplate-wire stapling spinal fixation for traumatic injuries in snakes was undertaken on ten snakes (six spectacled cobra and four rat snake) presented to the PFA (People for Animals) and WRRC (Wildlife Rescue ad Rehabilitation Centre). Vertebral fractures were more common in spectacled cobras, and the most common causes of traumatic spinal injuries were animal attacks, underground excavation work, automobile accidents, and human attacks. Radiography and neurological examination viz. righting, deep pain, panniculus and cloacal reflexes were successful in determining the fractured site, assessing the status of implants after surgery, and healing of vertebral fractures and luxations.The results of ten snakes treated with modified microplate-wire stapling spinal fixation, four of which showed clinical recovery by the end of the study . In traumatically injured snakes with vertebral fracture and luxation, modified microplate-wire stapling spinal fixation was found to be effective. This method was found to be easy to use and provided rigid fixation. It was simple to apply in the dorsolateral region without requiring advanced surgical skills, and snakes were relatively comfortable because it provided symmetrical fixation close to the spine's neutral axis. Physiological parameters did not show any significant changes (rectal temperature, respiratory rate and heart rate). Neurological examination showed significant improvement in the reflexes. In this technique, post-operative complications were kept to a minimum. This technique efficiently provided a good stability at the fractured site in traumatic spinal injured snakes and had good success rate (40%).ThesisItem Open Access EFFECT OF PROBIOTICS IN CORN REPLACED RICE BASED DIET IN BROILER CHICKEN(COLLEGE OF VETERINARY AND ANIMAL SCIENCES MANNUTHY, THRISSUR, KERALA VETERINARY AND ANIMAL SCIENCES UNIVERSITY, 2023-03-23) RESHMA RAVEENDRAN; Dr. Binoj ChackoAn experiment was conducted in the Department of Poultry Science, College of Veterinary and Animal Sciences, Mannuthy to find out the effect of probiotics in corn replaced rice based diet in broiler chicken for a period of 42 days. A total of 144, day-old Ven Cobb broiler chicks were allotted randomly to six dietary treatment groups with three replicates of eight birds each. The treatment groups were T1-standard broiler ration (SBR1), T2- SBR1 with 0.05 per cent probiotic mixture, T3- SBR1 with 0.1 per cent probiotic mixture, T4- 50 per cent of maize from SBR1 replaced by rice (SBR2), T5- SBR2 with 0.05 per cent probiotic mixture, T6- SBR2 with 0.1 per cent probiotic mixture. The body weight, body weight gain, feed consumption and carcass characteristics were not affected by the inclusion of rice and probiotics in broiler pre-starter and starter diets. The feed efficiency was best in broilers fed rice without probiotic supplementation. Whereas, the probiotic supplementation in rice based diet resulted in poorer FCR. The dietary inclusion of rice did not affect the E. coli, coliform count and ileal pH. Whereas, the E. faecalis count was higher in rice fed group than corn fed group. The probiotics supplementation at 0.1 per cent level significantly (p<0.01) reduced the E. coli, coliform, E. faecalis count and ileal pH both in rice and corn based diets. The dietary inclusion of rice in broilers improved the ileal villi height and VH:CD ratio. The supplementation of probiotics in both rice and corn based diets significantly (p<0.01) increased the villi height of duodenum, jejunum and ileum and VH:CD ratio in jejunum and ileum. The dietary inclusion of rice did not affect the serum total cholesterol, triglycerides, HDL, LDL and VLDL value in broilers. Whereas, the supplementation of probiotics significantly (p<0.01) reduced the total cholesterol, triglycerides and VLDL in both rice and corn based diets. The serum triglyceride and VLDL level was significantly (p<0.01) lower in birds fed on rice diet with probiotics than birds fed corn based diet with probiotics. The dry matter, crude protein and ether extract digestibility of groups fed on rice based diet was better compared to groups fed on corn diet. The protein digestibility was significantly improved by probiotic supplementation at 0.1 per cent level in corn diet. There was no such effect of probiotics in rice fed groups. The crude protein and ether extract digestibility in rice with 0.05 per cent probiotics group was better than corn with 0.05 per cent probiotics fed group. The crude fibre digestibility was not affected by inclusion of rice or probiotics in broiler diet. The cost of feed per kg body weight was lowest in group fed rice without probiotic supplementation. The supplementation of probiotics in corn based diet resulted in lower feed cost per kg body weight. No such effect of probiotics was observed in rice based diet. In conclusion, replacement of 50 per cent of corn by rice from the diet and supplementation of diet with probiotics at 0.05 per cent level can be recommended during pre-starter and starter phase of broilers.ThesisItem Open Access EVALUATION OF BENZIMIDAZOLE RESISTANCE IN GASTROINTESTINAL NEMATODES OF CATTLE(COLLEGE OF VETERINARY AND ANIMAL SCIENCES MANNUTHY, THRISSUR, KERALA VETERINARY AND ANIMAL SCIENCES UNIVERSITY, 2023-03-23) MERIN DAS V. J; Dr. Priya M. NThe present study was carried out to evaluate benzimidazole resistance in gastrointestinal nematodes of cattle. A total of 497 faecal samples were collected from 17 organised cattle farms and 23 small holder farmers’ herd in Thrissur district and adjoining areas. Microscopical examination of the faecal samples revealed an overall incidence of 36.01 per cent of gastrointestinal parasitism in cattle. The incidence of strongylosis among the samples examined was 10.66 per cent with incidence rates of 6.94 and 17.22 per cent in organised and small holder farmers’ herd respectively. There was no significant difference between mean incidence of strongyles in organised farm (11.34±5.60 per cent) and that of small holder farmers’ herd (22.95±4.83). However, there was a significant difference between mean faecal egg count (FEC) in organised farm (1559.09±731.789) and that of small holder farmers’ herd (134.38±27.719). The prevalent genera of strongyles identified by coproculture were Haemonchus spp., Mecistocirrus spp. and Oesophagostomum spp. Detection of anthelmintic resistance was done by in vivo faecal egg count reduction method (FECRT), in vitro egg hatch assay (EHA) and molecular genotyping by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). Based on FECRT with albendazole, resistance was detected in all organised farms and 33.33 per cent of the small holder farmers’ herd whereas 66.67 per cent of the small holder farmers’ herd were susceptible to benzimidazole. Pooled samples from three organised and one small holder farmers’ herd were utilised for EHA and resistance was observed in two organised farms and one small holder farmers’ herd with ED50 greater than 0.1µg/mL and Hdd greater than 0.5 µg/mL whereas one organised farm was detected as susceptible to benzimidazole. Molecular detection by PCR-RFLP was done by using single larvae of Haemonchus spp. from pooled faecal sample of three organised and one small holder farmers’ herd followed by restriction digestion with Eco1051, HpyCH4V and HpyCh4III to determine resistance associated SNPs at codon 167, 198 and 200, respectively at isotype 1 β- tubulin gene. The results of PCR-RFLP with 102 Haemonchus spp. larvae revealed 95.10per cent of the larvae carried resistant genotype (rr) and 4.9 per cent carried susceptible genotype (SS) at codon 198 of isotype 1 β tubulin gene. All the larvae carried susceptible genotype (SS) at codon 167 and 200 of the β tubulin gene. The association of clinical benzimidazole resistance by FECRT with molecular genotyping was done and revealed that benzimidazole resistant organised farms showed 97.7 per cent homozygous resistant genotype (rr) and 2.30 per cent homozygous susceptible genotype (SS). The benzimidazole resistant small holder farmers’ herd showed 81.25 per cent rr genotype and 18.75 per cent SS genotype. In conclusion, the present research work reports the detection of benzimidazole resistance in cattle by in vivo FECRT, in vitro EHA and molecular genotyping by PCR-RFLP to identify the SNPs at isotype 1 β- tubulin gene of Haemonchus spp. To the best of our knowledge, this study forms the first report of anthelmintic resistance in cattle in South India.ThesisItem Open Access ANTINEOPLASTIC POTENTIAL OF Magnolia champaca (CHEMBAKAM) AND Epipremnum aureum (MONEY PLANT) LEAVES IN MCF-7 CELL LINE(COLLEGE OF VETERINARY AND ANIMAL SCIENCES MANNUTHY, THRISSUR, KERALA VETERINARY AND ANIMAL SCIENCES UNIVERSITY, 2023-03-23) SHANKAR R; Dr. Preethy JohnCancer is ranked as the primary cause of mortality and a significant roadblock to raising life expectancy in every part of the world. Though being targeted by multimodal therapies, cancer is still the most common aetiology for loss of life worldwide. The adverse effects associated with conventional therapies and the development of resistance to them warrant investigation of alternative therapeutic agents. Thus, the present study was undertaken to evaluate the antineoplastic potential of the methanolic extracts of Magnolia champaca (MMC) and Epipremnum aureum (MEA) leaves in Michigan Cancer Foundation-7 (MCF-7) cell line and to compare the effect with doxorubicin. The leaves of M. champaca and E. aureum were collected, cleaned, shade dried, extracted using methanol with Soxhlet apparatus and stored until use. The phytochemical constituents were analysed qualitatively and with gas chromatography-tandem mass spectrometry (GC-MS/MS) and fourier transform infrared (FTIR) spectroscopy. The phytochemicals identified were screened for its activity using Prediction of Activity Spectra for Substances (PASS). Those compounds with antineoplastic activity were docked with Bcl-2 protein using Autodock4. The cytotoxic potential of MMC and MEA were assessed using MTT assay at the doses of 10,20,40, 80,160 and 320 μg/mL from which per cent cell viability, per cent inhibition and half-maximal inhibitory concentration (IC50) were calculated. Doxorubicin was used as a positive control. With the IC50concentration, the apoptotic changes were studied using dual acridine orange AO)-ethidium bromide (EB) staining. The ability of the extract to generate intracellular ROS in the cells were assessed using 2’,7’ dichlorofluoresceindiacetate (DCF-DA) assay. The effect of MMC and MEA on the expression of anti-apoptotic gene Bcl-2 was studied using real time - quantitative polymerase chain reaction (RT-qPCR) with glyceraldehyde-3-phosphate dehydrogenase(GAPDH) as house-keeping gene.The extraction yields of MMC and MEA were found to be 21.26 and 17.56 per cent respectively. The qualitative phytochemical screening revealed the presence of steroids, alkaloids, flavonoids and terpenes in both the extracts, in addition tannins, phenols and glycosides in MMC. The GC-MS/MS analysis revealed presence of 99 and 93 compounds in MMC and MEA respectively. The FTIR spectroscopic analysis revealed many peaks that depicted the presence of structurally related compounds and confirmed the presence of phytochemicals that were previously screened qualitatively. The in silico docking analysis revealed good interaction of many compounds with the Bcl-2 protein. Among the various compounds, the binding of ethyl iso-allocholate, β-copaene, ascaridole epoxide and caryophyllene oxide with the Bcl-2 protein were significant with a binding energy of -5.71, -5.7, -5.00 and -4.91 kcal/mol, respectively.The MTT assay revealed a dose-dependent inhibition of cell viability in both the extracts. Based on the per cent inhibition, the IC50 values of doxorubicin, MMC and MEA were calculated to be 15.27, 66.61 and 104.4 µg/ mL respectively. On AO/EB staining, the untreated control cells were viable that emitted uniform green fluorescence whereas cells treated with MMC and MEA revealed early apoptotic changes like nuclear condensation and marginalisation. Nuclear fragmentation and late apoptotic changes could be seen in doxorubicin cells. The DCF-DA assay revealed significant generation of ROS in doxorubicin treated cells whereas cells treated with MMC and MEA scavenged the basal ROS, decreasing their intensities. The RT-qPCR revealed, critically up regulation of Bcl-2 in untreated control cells, whereas a significant down regulation of the gene in cells treated with MMC (0.15-fold) and MEA (0.34-fold)was found which was comparable with the doxorubicin (0.23-fold). From the present study, it can be concluded that both MMC and MEA possessed antineoplastic activity that produced apoptosis-induced ROS independent cytotoxicity and MMC found to be comparatively more potent than MEA. However, further studies are essential for developing novel therapeutic candidates against cancer with M. champaca and E. aureum.ThesisItem Open Access EFFECT OF FERMENTED COTTONSEED MEAL ON GROWTH PERFORMANCE, GUT MICROFLORA AND CARCASS TRAITS IN BROILERS(COLLEGE OF VETERINARY AND ANIMAL SCIENCES MANNUTHY, THRISSUR, KERALA VETERINARY AND ANIMAL SCIENCES UNIVERSITY, 2023-03-23) ARUNDHATHI U; Dr. P. AnithaAn experiment was conducted at the Department of Poultry Science, College of Veterinary and Animal Sciences, Mannuthy to study the effect of dietary inclusion of fermented cottonseed meal (FCSM) on growth performance, intestinal microflora and carcass traits in broiler chicken up to 42 days of age. A total of 160, day-old chicks were randomly allotted into five dietary treatment groups, viz. T1, T2, T3, T4 and T5 with four replicates having eight bird each, in a completely randomised design. The birds in the control group (T1) were fed with standard broiler ration formulated as per BIS (2007). The diet groups T2 to T5 were formulated by inclusion of five per cent CSM (T2), five per cent FCSM (T3), 10 per cent FCSM (T4) and 15 per cent FCSM (T5), respectively. All the experimental diet were isocaloric and isonitrogenous. Microbial fermentation of CSM significantly (p<0.05) increased its moisture, crude protein and total ash content and also improved the lactobacillus count (LAB). The dry matter, crude fibre, free gossypol, aflatoxin B1 level and pH were significantly (p<0.05) lower in FCSM compared to non-fermented CSM. The mean body weight and body weight gain of broilers at the sixth week of age in different treatment groups did not differ significantly. The mean cumulative feed consumption of broilers at sixth week of age did not show any significant difference between the treatment groups. The cumulative mean feed conversion ratio from first to sixth week of age was not significantly affected by the dietary treatments. The carcass traits like pre slaughter live weight, eviscerated weight, abdominal fat, per cent weight of heart, liver and gizzard, yield of thigh, drumstick and wings (per cent) did not differ significantly among different treatment groups. The mean ready to cook yield (per cent) was significantly (p<0.05) higher in all FCSM groups (T3, T4 and T5) than the other two groups.The mean count of harmful bacterial organisms like Escherichia coli, coliform, and Enterococcus faecalis count of ileal content in broilers was significantly (p<0.01) lower in FCSM groups (T3, T4 and T5) as compared to control diet (T1) and five per cent CSM diet (T2). The mean ileal pH was also significantly (p<0.01) lower in FCSM groups (T3, T4 and T5) than the other two groups. The mean villi height, villi height to crypt depth ratio (VH:CD) in duodenum and jejunum was significantly (p<0.01) higher in FCSM groups. The mean villi height of ileum was significantly (p<0.01) higher and crypt depth of duodenum and jejunum was significantly (p<0.05) lower in FCSM groups compared to control and CSM group (T2).The serum total cholesterol, low-density lipoprotein and very low-density lipoprotein levels of birds at sixth week of age was significantly (p<0.01) lower in T3, T4 and T5 groups compared to control (T1) and CSM group (T2). The mean livability per cent in T1, T2, T3, T4 and T5 was in range of 90.62 to 100. The highest net profit per kilogram body weight was obtained in 10 per cent FCSM group (T4 group), followed by T3, T5, T2 and T1. Based on the overall findings of this study, it can be concluded that the dietary inclusion of fermented cottonseed meal at different levels by replacing soybean meal resulted in similar growth performance as that of the control group with the soybean meal diet. Therefore, FCSM in broiler diet up to 15 per cent inclusion can be recommended as an alternative protein to soybean meal without affecting the performance of the birds and with beneficial effect on the gut health. Among the different levels of inclusion studied, 10 per cent level was found to be more beneficial with respect to net profit per kilogram body weight.ThesisItem Open Access CLINICAL EVALUATION OF A MULTIMODAL ANAESTHETIC PROTOCOL USING BUTORPHANOL-DEXMEDETOMIDINE MIDAZOLAM-KETAMINE-ISOFLURANE AND LIGNOCAINE CONTINUOUS RATE INFUSION IN CATTLE(COLLEGE OF VETERINARY AND ANIMAL SCIENCES MANNUTHY, THRISSUR, KERALA VETERINARY AND ANIMAL SCIENCES UNIVERSITY, 2023-03-23) JAYAKRISHNAN V; Dr. Sudheesh S. NairThe present study was conducted for clinical evaluation of a multimodal anaesthetic protocol using butorphanol-dexmedetomidine-midazolam-ketamine isoflurane and lignocaine continuous rate infusion in six cross-bred female cattle with mean weight of 333.00 ± 46.32 kg. Pre-anesthetics used were butorphanol and dexmedetomidine which were administered intravenously at dose rates of 0.05 and 1 µg/kg body weight, respectively. Moderate quality sedation was achieved within a mean time of 7.67 ± 1.08 minutes. Ketamine and midazolam were intravenously administered at dose rates of 4.0 and 0.2 mg/kg body weight respectively, to induce anaesthesia. Excellent quality induction was achieved within a mean time of 5.83 ± 1.11 minutes. Isoflurane was used to maintain anaesthesia after endotracheal intubation, at a concentration of 1.056 ± 0.97 per cent concentration in 100 per centoxygen using a large animal anaesthesia machine. Simultaneously, intravenous lignocaine was administered at a bolus dose of 2 mg/kg followed by a constant rate infusion of 3 mg/kg body weight using a volumetric infusion pump. Isoflurane sparing effect of intravenous lignocaine reduced the required per cent of isoflurane. The third plane of surgical anaesthesia was maintained and various surgical procedures were done. During the course of anaesthesia, the differences in heart rate, rate of respiration and rectal temperature were found to be statistically non-significant. Simultaneously, a significant rise in capillary refill time was also noticed. Haematological, serum biochemical, blood gas and electrolyte parameters recorded non-significant changes. Recovery was smooth and the mean time required to exhibit the first voluntary reflex,assume sternal recumbency and stand up unassisted following the cessation of isoflurane anaesthesia were 7.50 ± 1.82 minutes, 15.17 ± 5.16 minutes and 35.33 ± 17.04 minutes respectively. Other than the mild regurgitation of ruminal fluid in an animal, no anaesthetic complications were noticed.ThesisItem Open Access DEVELOPMENT AND EVALUATION OF ENROBED CHICKEN NUGGETS INCORPORTAED WITH MILLETS AND HYDROCOLLOIDS(COLLEGE OF VETERINARY AND ANIMAL SCIENCES MANNUTHY, THRISSUR, KERALA VETERINARY AND ANIMAL SCIENCES UNIVERSITY, 2023-03-23) AKHILA V. V.; Dr. Sathu TThe present study on “Development and evaluation of enrobed chicken nuggets incorporating millets and hydrocolloids” was conducted in the Department of Livestock Products Technology, College of Veterinary and Animal Sciences, Mannuthy. The study was conducted for the development of enrobed chicken nuggets incorporated with millet flour in the nuggets and hydrocolloid in the nuggets enrobing batter. The study was conducted in four phases, viz., 1. Standardisation of the formulation of enrobed chicken nuggets, 2. Selection and standardisation of level of millet in chicken nuggets, 3. Selection and level of incorporation of hydrocolloid in chicken nugget enrobing batter, 4. Studies on shelf life of the standardised product. The standardised enrobed chicken nuggets was incorporated with pearl millet (3.5%) in nuggets and xanthan gum (1%) in the nugget enrobing batter. The standardised product contains 43.44 % moisture, 14.75% protein, 15.02 % fat whereas the control sample contains 41.73%, 19.56 % and 17.67 % respectively. The control and the standardised product were packed aerobically in a laminated pouches and stored under refrigeration condition (4±1ºC) for shelf life study. Physicochemical parameters, TBARS value, tyrosine value, colour, microbiological assessment and sensory evaluation were performed on 0th, 5th, 10th and 15th day of storage period. The pH and water activity of the control and the standardised product significantly (p<0.001) decreased during storage days. TBARS and tyrosine value was increasing throughout the storage period and also significantly(p<0.001) differed during storage, TBARS value of control ranged from 0.26 to 0.41mg malonaldehyde/kg and from 0.25 to 0.36 mg malonaldehyde/kg for standardised product. The tyrosine value ranged from 4.62 to 12.78 for control and 3.96 to 12.78 for standardised product. Significant difference (p<0.001) was noticed between samples for the microbiological count (aerobic plate count and yeast and mould count). As the storage period advanced the aerobic plate count of the standardised product increased from log10 2.85±0.07 CFU/g to log104.03±0.02 CFU/g and control from log10 2.68 ±0.1 CFU/g to log103.9 ±0.03 CFU/g. Evaluation of the sensory attributes during the shelf life study had significant difference (p<0.001) between the standardised product and control and also between the days as the storage period advances (p<0.001). From the above studies it can be concluded enrobed chicken nuggets cab be prepared with good acceptability by incorporating pearl millet flour (3.5%) in nuggets and xanthan gum (1%) in the enrobing batter.ThesisItem Open Access EVALUATION OF VITAMIN AND MINERAL STATUS IN CHRONIC SKELETAL DISORDERS IN DOGS(COLLEGE OF VETERINARY AND ANIMAL SCIENCES MANNUTHY, THRISSUR, KERALA VETERINARY AND ANIMAL SCIENCES UNIVERSITY, 2023-03-23) RADHIKA R; Dr. Aziz ZarinaDogs in their early growth phase are more susceptible to nutritional deficiency diseases. The bone health is monitored by both water-soluble and fat- soluble vitamins, along with calcium and phosphorus. Fast growing dogs with unbalanced diets are at higher risk of developing skeletal abnormalities due to increased metabolic activity and lower levels of serum vitamins and minerals. The present study was undertaken to evaluate the vitamin and mineral status in chronic skeletal disorders in dogs. In this study, 30 large breed dogs below 18 months with chronic skeletal disorders presented to the University Veterinary Hospitals at Mannuthy and Kokkala under the Kerala Veterinary and Animal Sciences University were selected and ten apparently healthy dogs brought to the hospital for routine health-check up without any symptoms of skeletal deformities served as control. Blood was collected from these animals to analyse serum biochemical parameters. The serum concentrations of water-soluble vitamins (B1, B2, B3, B6, B12 and vitamin C) and cholecalciferol were analysed by ultra-high performance liquid chromatography (HPLC). The serum calcium and magnesium were analysed by atomic absorption spectrophotometry (AAS) and levels of serum alkaline phosphatase, acid phosphatase and phosphorus were estimated by fully automated biochemical analyzer. The levels of serum homocysteine were determined spectrophotometrically by enzyme linked immune-sorbent assay (ELISA). The levels of serum vitamin B2, B6, B12, C and D3 were significantly (p<0.01) lower in dogs with chronic skeletal disorders compared to control dogs. However, serum vitamin B1 and B3 did not differ significantly in dogs with chronic skeletal disorders and control dogs. The serum concentrations of calcium, phosphorus and magnesium were significantly (p<0.05) lower in affected dogs compared to control dogs. A significant (p<0.01) increase in serum alkaline phosphatase, acid phosphatase and homocysteine levels were observed in dogs affected with chronic bone abnormalities compared to normal. The results of this study indicated that decreased levels of serum vitamins (B2, B6, B12, C and D3), minerals (calcium, phosphorus and magnesium) and increased levels of serum alkaline phosphatase, acid phosphatase and homocysteine contributed to development of chronic skeletal disorders in growing dogs.
