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Open Access
Evaluation of hepatoprotective effect of ethanolic extract of eugenia jambolana (njaval) leaves on paracetamol induced toxicity in rats
(Department of Pharmacology and Toxicology, College of Veterinary and Animal Sciences Mannuthy, 2008) Midhun, M V; Aravindakshan, C M
Open Access
Comparative evaluation of membrane protein and biofilm vaccines against duck pasteurellosis
(Department Of Poultry Sciences,College of Veterinary and Animal Sciences, Mannuthy, 2008) Indu, K; KAU; Krishnan Nair, C
A research work was undertaken to prepare effective vaccines against P. multocida grown under different conditions and assess their immunopotency in one month old ducklings. The purity of the Pasteurella multocida A: 1 strain (DP1) and A: 4 strain (PA4) was confirmed as per standard procedures. Pathogenicity of DP1 and PA4 was assessed in six to eight week old mice. The isolates killed the intraperitoneally inoculated mice within eight hours and within 24 h when injected by subcutaneous route. Pasteurella multocida A: 1 was used for the preparation of different vaccines. The organism was grown in BHIB for preparation of ordinary bacterin. The in vitro biofilm formation of the organism was assessed by growing it under nutrient restricted conditions. For this, the organism was grown in TSB (0.32 per cent) supplemented with 0.3 per cent bentonite clay. For preparation of OMP suspension, the organism was grown in iron restricted condition viz., BHIB supplemented with 100 µM 2, 2’ Dipyridyl and the OMPs were extracted using sodium lauryl sarcosinate. The protein concentration of OMP suspension was estimated to be 3 mg/ml. Median lethal dose (LD50) of DP1 was 10-7.5, which contained 32 viable cells/ ml and that of PA4 was 10-7.38, which contained 24 viable cells/ ml when determined in one month old ducklings. Oil adjuvant vaccines were prepared using ordinary bacterin, bacterin made from biofilm and OMP suspension and performed the sterility, safety and potency tests of the vaccine employing standard procedures. A total of 260 four week old ducklings were divided into four groups with 65 birds in each group and the first three groups were vaccinated with ordinary bacterin, OMP vaccine 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 3 weeks post vaccination (PV) and then at 15 days interval upto 60 days, 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. On homologous challenging, biofilm vaccine gave higher protection rates of 80 per cent than the 70 and 50 per cent protection rates of ordinary bacterin and OMP vaccine respectively, when challenged with 100 LD50 dose on day 20 PV. On day 60 PV, biofilm vaccine gave higher protection rate of 70 per cent than the 60 and 50 per cent protection rates respectively of ordinary bacterin and OMP vaccine, when challenged with 100 LD50 dose. On heterologous challenging, biofilm vaccine gave higher protection rates of 70 per cent, while only 50 per cent protection was afforded by both bacterin and OMP vaccine, when challenged with 100 LD50 dose on 20 day PV. On day 60 PV, biofilm vaccine gave higher protection rate of 60 per cent while both the other vaccines gave only 50 per cent protection, when challenged with 100 LD50 dose. All the birds challenged on day 40 PV, either with homologous and heterologous organisms died. In most cases, the death occurred due to coliform infection along with stressful factor such as increased atmospheric humidity due to heavy rainfall at that time. In few cases, birds died due to pasteurellosis which might be due to lack of protective level of antibody titre. Biofilm vaccine proved to be the best among the three vaccines tried. Further field trials are to be done before advocating this vaccine for commercial use.
Open Access
Effect of blood urea nitrogen,mineral status and uterine PH on fertility in dairy cows
(Department of Animal Reproduction, Gynaecology and Obstetrics, College of Veterinary and Animal Sciences, Mannuthy, 2008) Seena, N S; KAU; Athman, K V
An investigation was carried out with the objective of studying the effect of BUN and uterine pH on fertility in dairy cows under farm and field conditions and also for correlating the mineral status with fertility using 40 crossbred dairy cows selected at random during oestrus, 20 each from those belonging to University Livestock Farm, Mannuthy (Group I) and those brought for insemination at Artificial Insemination Centres at Mannuthy and Kokkalai (Group II). All the cows in Group I were found to be maintained in a relatively high nutritious diet computed as per the scientific feeding standards compared to Group II animals. Detailed clinico-gynaecological examination was carried out and blood samples and uterine mucus were collected from all the selected cows during oestrus for estimation of biochemical parameters and uterine pH respectively. They were inseminated during the most appropriate period of oestrus and were subjected to pregnancy diagnosis at 60 days post insemination. Conception rates in both the groups were compared in relation to each parameter. The mean duration of oestrus was slightly higher in Group II (30.00 ± 2.11 hours) compared to Group I (26.10 ± 1.74 hours). Intensity of oestrus was high, medium and low in 40, 45 and 15 per cent of animals respectively in Group I and 50, 40 and 10 per cent in Group II. Physical changes of reproductive tract viz. degree of vulval oedema and hyperemia of vestibular mucous membrane were more pronounced in animals of Group II compared to Group I, where as degree of tonicity was high in Group I compared to Group II. Characteristics of cervical mucus were also found to be affecting fertility. Better conception rate was obtained in animals with clear and stringy cervical mucus exhibiting typical type of fern pattern. Spinnbarkeit value did not vary much between conceived and non-conceived animals. Uterine pH did not show a marked variation between groups, even though a slightly higher value was recorded in Group I. But, an inverse relationship could be obtained between BUN level in serum and uterine pH during oestrus. Correlation between uterine pH and BUN level was highly significant (P<0.01) with a correlation co-efficient of r = -0.896 and r = - 0.753 in groups I and II respectively. The mean blood urea nitrogen level in animals of Group I was significantly higher (P<0.05) than that in Group II. The BUN level also varied significantly (P<0.05) between conceived and non-conceived animals of both groups. A marginal increase in plasma glucose level could be noticed in conceived animals compared to non-conceived animals of Group I. The mean serum total protein was significantly higher (P<0.01) in Group I compared to Group II. The serum level of minerals viz. calcium, phosphorus, manganese, zinc and copper were also correlated with fertility. The mean level of serum calcium and manganese varied significantly between groups I and II, but there was no significant difference in serum phosphorus, zinc and copper between two groups. Also, a slightly higher mean value was observed for serum calcium, phosphorus, manganese and zinc in conceived animals compared to non-conceived. But serum copper level did not vary between conceived and non-conceived animals. In light of these findings, it can be concluded that the elevation in systemic concentration of urea is likely to impair fertility in dairy cows as a consequence of alterations in uterine environment. The benefits of feeding excess dietary protein and urea to maintain peak milk production should be compared with potential negative effects on fertility. Hence, a good nutritional management is essential for improved fertility in dairy cows.
Open Access
Effect of gonadotropin releasing hormone and prostaglandin for improving reproductive efficiency in goats
(Department of Animal Reproduction, Gynaecology and Obstetrics, College of Veterinary and Animal Sciences, Mannuthy, 2008) Julliet; KAU; Joseph, Mathew
With the objective of studying the effect of GnRH and prostaglandin for improving reproductive efficiency in goats the study was carried out at University Sheep and Goat Farm, Mannuthy using 42 cycling goats. Based on the behavioural and physiological changes associated with oestrum the goats were divided into two groups viz., Group I and Group II. Group I animals were those that exhibited pronounced oestrus signs and were divided into two subgroups namely Group IA and Group IB. Group II animals were those that exhibited weak oestrus signs and were divided into three subgroups namely Group IIA, IIB and IIC. Group IA animals were administered 0.0042 mg Buserelin (1 ml Receptal) a potent GnRH analogue on day 0, and Group IB served as the Control. Blood was collected prior to GnRH administration and breeding from all does. The mean duration of oestrum in Group IA and IB was 19.33 ± 0.45 and 33 ± 0.58 h respectively. The conception rate in Group IA and IB was 50 per cent and 66.66 per cent respectively. The serum P4 level on day 0 in does in Group IA and IB was 0.43 ± 0.05 ng/ml and 0.40 ± 0.05 ng/ml respectively. Group IIA and Group IIB does were treated as per the CO-Synch protocol (i/m inj. of 0.0042 mg of Buserelin (1 ml Receptal) on day 0, 125 µg cloprostenol (0.5 ml clostenol) on day 7; 0.0042 mg of Buserelin and mating on day 9) and prostaglandin protocol respectively (two intramuscular injections of 125 µg cloprostenol (0.5 ml clostenol) 11 days apart followed by mating at 72 and 96 h), while Group IIC served as the control. The oestrus response, oestrus onset interval, duration of oestrum and conception rate in Group IIA was 90.9 per cent, 47.6 ± 0.45 h, 24.5 ± 0.63 h and 40 per cent respectively. The oestrus intensity score of induced oestrus ranged from 0 to 13. The serum P4 level in pregnant and non pregnant does was not significantly different on days 0, 7 and 9 (P>0.05). The oestrus response, oestrus onset interval, duration of oestrum and conception rate in Group IIB was 81.8 per cent, 54 ± 1.006 h, 39.77 ± 1.54 h and 66.66 per cent respectively. The oestrus intensity scores in induced oestrus ranged from 0 to 13. The serum progesterone level in does that became pregnant and those that were non pregnant were not significantly different on day 0, 11, and at 72 and 96 h. In Group II C the duration of oestrum and pregnancy rates was 40 ± 0.91 h and 33.33 per cent respectively. Pregnancy diagnosis was done at three months of gestation by abdominal palpation and the accuracy of the method was 90.9 per cent. Mean gestation length was 146.03 ± 0.76 days. Litter size at birth in Group IA, IB, IIA, IIB and IIC was 2, 2, 2, 1.83 and 2 respectively. Average birth weight of kids was 2.35 ± 0.164 kg and the mean birthweight of male and female kid was 2.42 ± 0.98 kg and 2.28 ± 0.36 kg respectively. Thus from the present study, it can be concluded that :- 1. Administration of GnRH on the day of oestrum in animals exhibiting pronounced oestrus signs failed to improve conception rate when compared to the control. 2. In animals exhibiting weak oestrus signs both CO-Synch and double prostaglandin protocols resulted in higher conception rate when compared to control group. 3. The double prostaglandin protocol was found to be more efficient in improving conception rate in animals exhibiting weak oestrus signs.
Open Access
Surveillance of haemoprotozoan and haemorickettsial diseases of cattle of northern Kerala
(Department Of Veterinary Epidemiology & Preventive Medicines,College of Veterinary And Animal Sciences, Mannuthy, 2008) Archana Nair, S; KAU; Tresamol, P V
A cross-sectional study was conducted to assess the carrier status of haemoprotozoan and haemorickettsial diseases of cattle of Northern Kerala. The diagnostic methods employed were microscopical examination of blood smears stained with Giemsa and Acridine Orange and polymerase chain reaction. A total of 150 blood samples were collected from the five districts of Northern Kerala (30 samples from each district). Microscopical examination of Giemsa stained smears revealed Theileria like piroplasms (41.2 per cent), Babesia piroplasms (2.6 per cent) and Anaplasma inclusions (1.3 per cent) while Acridine Orange stained smears detected Theileria like piroplasms in 47.2 per cent, Babesia piroplasms in 2.6 per cent and Anaplasma inclusions in 2 per cent of blood smears examined. Acridine Orange staining is more easy, time saving and reliable in detecting of haemoparasites than Giemsa staining technique. Polymerase chain reaction detected haemoprotozoan like T. evansi (34.6 per cent), B. bigemina (0.6 per cent), an uncharacterized Theileria organism (16 per cent) and T. orientalis (1.3 per cent). A. marginale (16.7 per cent) and A. bovis (3.3 per cent) were the haemorickettsial organisms detected by PCR. Mixed infections were detected in 16.6 per cent of the samples. T.annulata, B. bovis and A. phagocytophila were not detected in any of the samples. Polymerase chain reaction showed much higher sensitivity compared to examination of stained smears in detecting carrier animals. Trypanosomosis is the most prevalent protozoan disease among carrier animals of Northern Kerala which were detected only with the help of PCR while microscopy totally failed to detect such carriers. The high prevalence of haemoprotozoan and haemorickettsial diseases suggests the abundance of ticks and flies in Northern Kerala which acts as vectors for these parasites. Kannur and Palakkad districts revealed maximum prevalence of T. evansi. In addition, anaplasmosis was also widely prevalent in Palakkad district. Microscopical examination of blood smears from cattle of the hilly districts of Wayanad, Malappuram and Kozhikode revealed the presence of a Theileria like piroplasm which were not amplified with PCRs specific for Theileria genus, T. annulata or T. orientalis and this uncharacterized piroplasm needs urgent attention.
Open Access
Energy supplementation on production performance of cows under field condition
(Department of Animal Nutrition, College of Veterinary and Animal Sciences, Mannuthy, 2008) Ruby, Augustian; KAU; Mercy, A D
A study was conducted to assess the effect of energy supplementation in the form of maize and rumen protected fat in early lactating cows under the field condition. Thirty, early lactating cross bred cows were selected and divided into three groups uniformly with regard to days in milk, the average days in milk being 40 days. The animals were randomly allotted to the dietary treatments T1, T2 and T3. The concentrate mix made in the semi liquid form and paddy straw formed the T1 ration. The animals of T2 and T3 were supplemented with 1 kg ground maize and 100 g of rumen protected fat respectively, over and above the control ration. The animals maintained on the three dietary treatments T1, T2 and T3 showed an average milk production of 15.38, 14.58 and 13.01 kg respectively during the 90 day period of the experiment. Statistical analysis showed no significant difference (P>0.05) in milk yield between the three treatment groups. The average daily DMI and DMI/100 kg body weight remained similar for all the three groups. The TDN intake was up to the requirement in animals maintained on T3 ration while T1 and T2 rations could not meet the requirements of the animals of the respective groups. The DCP intake was optimum from the three rations. The energy supplementation did not affect milk composition such as percentage of total solids, milk fat, solids not fat (SNF) and milk protein, and milk urea nitrogen (MUN). The 4 per cent FCM yield, total yield of milk fat and protein remained unaffected by energy supplementation. Animals in all the three groups showed similar blood parameters such as haemoglobin, plasma glucose, plasma urea nitrogen and plasma Ca and P. The digestibility of EE, DM, NDF and ADF was higher for the T3 ration than that of T1 and T2. The digestibility of crude protein (CP) was higher for T2 than T1 and T3 rations. Animals maintained on T3 ration showed earlier post partum heat. The cost of feed for producing 1 kg milk was higher for the dietary treatments T2 and T3. An overall assessment of the results obtained in the present study showed that energy supplementation in the form of either maize or rumen protected fat could not influence dry matter intake, milk yield and 4 per cent FCM yield. There was no change in the composition of milk and blood parameters studied, among the experimental animals. However, energy supplementation in the form of protected fat could meet the energy requirement of early lactating animals, improved the digestibility of EE, DM, NDF and ADF and positively influenced the first post partum heat.
Open Access
Gastrointestinal and neurotoxic effects of cypermethrin in rats
(Centre For Excellence In Pathology, College of Veterinary and Animal Sciences, Mannuthy, 2008) Remya Nair, R; KAU
The present study entitled ‘Gastrointestinal and neurotoxic effects of cypermethrin in rats’ was undertaken to investigate the potential toxic effects of cypermethrin in rats with particular emphasis on its effect on the nervous and digestive system.The clinical signs, gross pathology, histopathology, haematology and biochemical parameters were analysed to study the effects. Cypermethrin was found to be neurotoxic and gastroenterotoxic at the given dose levels. Oral administration of cypermethrin in medium and high doses produced nervous signs in animals. However cypermethrin did not cause any significant variation in the body weight of animals. All haematological parameters evaluated showed a dose dependent reduction in its value. Biochemical parameters assessed revealed hepatotoxic and multisystemic effects of cypermethrin. Gross lesions observed in the intoxicated groups were bloat, congestion of lungs, heart, brain, pulmonary haemorrhage and degenerative changes in liver and kidneys. On histopathological examination, cypermethrin was found to be neurotoxic, hepatotoxic, enterotoxic, pneumotoxic, cardiotoxic and nephrotoxic as evidenced by varying degrees of degeneration and necrosis in various organs examined. Effects on all organs were mild to moderate degenerative changes at the low dose level. Medium and high dose intoxicated groups revealed necrotic changes, extensive haemorrhages, congestion in organs like liver, kidney and lungs apart from the changes observed in low dose group animals. Haemodynamic disturbances were manifested in the forms of congestion and haemorhages in lungs, heart, liver and kidneys. The histopathological evaluation revealed no cypermethrin induced toxic effects on the oesophagus. Spinal cord lesions were observed only in the lowest dose group. Brain lesions were only degenerative changes. These findings suggested that biochemical effects rather than structural changes were involved in toxicity.
Open Access
Comparative efficacy of diagnostic tests for paratuberculosis in goats
(Department of Veterinery Edodomology and Preventive Medicine, College of Veterinary and Animal Science, Mannuthy, 2008) Sulficar, S; KAU; Saseendranath, M R
Paratuberculosis (Johne’s disease) is a chronic debilitating infection of goats caused by Mycobacterium avium subsp paratuberculosis (MAP). Recently the organism was reported to be associated with enteric infection in humans and hence the disease is of public health importance. In the present study comparative efficacy of acid fast staining of faecal smear, single intradermal Johnin test and IS900 faecal PCR were investigated in two hundred goats for detection of MAP. Single intradermal Johnin test was carried out in two hundred goats. Faecal sample from all two hundred goats subjected to Ziehl-Neelsen acid fast staining and IS900 PCR for detection of MAP. Out of one hundred and fifty goats screened for paratuberculosis from University sheep and goat farm, Mannuthy, five animals (3.33 per cent) were found positive by acid fast staining, eight animals (5.3 per cent) were found positive by single intradermal Johnin test and thirty animals (20 per cent) were found positive by IS900 PCR. Out of fifty goats screened for paratuberculosis from field, one goat (2 per cent), three goats (6 per cent) and twelve animals (24 per cent) were found positive by Ziehl-Neelsen acid fast staining of faecal smear, single intradermal Johnin test and IS900 PCR respectively Results of present study from both farm and field indicate that amplification of the IS900 insertion element was the most specific and sensitive diagnostic method. The strategic use of PCR can provide a means for early identification of MAP infected goat, thus ensuring their elimination from an infected herd. Advantage of skin test is its ease of application and its low cost. Culling of CMI test positives could be a cost effective means of removing infected animals before they actually start faecal shedding. Ziehl-Neelsen staining had the lowest detection rate of the three tests under investigation and it proved unreliable in diagnosing Johne’s disease.
Open Access
Application of polymerase chain reaction for rapid evaluation of hygienic status of milk
(Department of Veterinary Public Health, College of Veterinary and Animal Sciences,Mannuthy, 2008) Deepa Mary, J J; KAU; Sunil, B
Rapid assessment of the bacterial load and detection of pathogens in milk is of public health significance. Molecular detection of pathogenic microorganisms is based on DNA amplification of the target pathogens. Therefore efficient extraction of DNA directly from milk is a major step. DNA could be efficiently extracted directly from milk by a prior sample preparation so as to remove the fat and milk proteins. The phenol chloroform method of DNA extraction was modified to reduce the time require for the procedure. The use of lysozyme helped the release of DNA from lysed gram positive Staphylococcus aureus. The extracted DNA was used as template in PCR. PCR was carried out with already published primers. PCR was modified with the use of PCR buffer containing PCR facilitators (BSA and Tween 20) to overcome PCR inhibition. The standardized procedure was used to assess the bacterial load and to detect Escherichia coli and S. aureus directly from milk. To assess the bacterial load dilutions of milk were made upto10-10. DNA was extracted from each dilution with which PCR was carried out with primers specific for Pseudomonas. Aerobic Plate Count was also done for the same samples and compared with PCR. It could be concluded that the approximate APC of the milk sample by PCR is next lower dilution to the dilution giving the PCR amplification. The total time taken for the analysis was approximately five hours. Extraction of DNA and PCR was done with primers for detection of E. coli from the same milk samples and compared with culture. Percentage of samples positive both by culture and PCR was 50 and negative by both methods were 30. Twenty percentage of the samples were positive by PCR and negative by culture. Extraction of DNA and PCR was done with primers for detection of S. aureus from the same milk samples and compared with culture. Percentage of samples positive both by culture and PCR was 60 and negative by both methods were 20. Twenty percentage of the samples were negative by PCR and positive by culture. Hence, protocol developed for detection of S. aureus needs further refinement to take care of false negative results by PCR, probably due to the low number of organisms present in milk.

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