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Tissue Research » Past Abstracts
1. Journal of  Cell and Tissue Research 15(2): 4929- 4942  (2015)
Ultrastructure and histopathological changes in testes and accessory glands in callosobruchus maculatus (f) (coleoptera, bruchidae) transpired by solar radiation

Khaled, A.S., Mohamed, M.I., Abdel Fattah, H.M., Hussein, M.A., Salem, D.A.M. and Fawki, S.
Entomology Department, Faculty of Science, Ain Shams University, Abbasiya 11566, Cairo, Egypt.
E. mail:,
Abstract: The ultrastructure and histopathology in the testes and accessory glands were studied in normal adult males and in those developed from solar energy treated pupae (53 ºC for 15 minutes) of Callosobruchus maculatus. The adult male reproductive system consists of two pairs of testes, two vasa deferentia, vesicular seminales, an ejaculatory duct and four pairs of accessory glands. Testes are formed of numerous testicular follicles containing germ cells at different stages of development. The transformation process of spermatid to sperm involves several morphological reorganization of the cells. Cross sections of elongating sperm tail of mature spermatozoa revealed the existence of two equal mitochondria derivatives containing crystalline material and the axial filament comprises 9+9+2 microtubules. C. maculatus has four pairs of accessory glands (one pair of lateral gland, the mesadenia and three pairs of median glands the ectadenia). The accessory glands consist of glandular cells with a well developed rough endoplasmic reticulum and Golgi apparatus. Variable morphological appearance of the accessory glands secretion was present in the gland lumen. The testicular follicles of treated pupae showed structural abnormalities. The cyst cells have a severe vacuolation, degeneration of germ cells, malformation and tearing of sperm bundles. Accessory glands were the most obvious signs of damage. The three different parts of the accessory glands (muscle layer, gland epithelium and lumen) changed as a result of solar effect. Deterioration of muscular layer, cytoplasmic organelles were very poor and the glandular lumen was filled with vacuoles and malformed granules. The damage was more pronounced in the gonads of (F) progeny of the treated generation.
 Key words: Callosobruchus maculatus, Male reproductive organs, Ultrastructure, Solar energy.
2. Journal of Cell and Tissue Research 15(2): 4943- 4947 (2015)
Effect of various solute concentrations on the membrane permeability of RBCS in human blood
Meena, Y., Romsha, S., Sneha, Y., Yadav, S. and Dhiman, S.
Department of Zoology, Maitreyi College, University of Delhi, Chanakyapuri, New Delhi 110021
E. mail:, Cell: 09818400124
Abstract: The development in the field of medicine has been immense in the past few decades. As the rate of diseases in humans as well as accidents is increasing, there is a constant need of the blood for the transfusion. The number of blood donors is limited, so the blood needs to be stored. A great care is needed to preserve the blood components specially RBCs. The integrity and physiology of the RBCs must be maintained to tackle the demand of blood in hospitals. A great care has to be taken to choose the possible donors with healthy blood. An increased level of the normal constituents may be harmful for the integrity of the RBCs. In the present study, the effect of abnormal levels of glucose, acetone, urea as well as saline on the membrane permeability of the RBCs was considered. These components i.e. 0.2 M glucose, 0.4 M glucose, 4% acetone, 5% acetone, 0.5% urea, 2% urea and hypotonic saline solutions, were shown to promote membrane permeability in RBCs, once RBCs were placed in them. However, the RBC membrane permeability was also found to be influenced by the temperature at which the RBCs were kept. Some solutions like 0.2% saline at 20°C, 5% saline at 20°C and 4% acetone at 20°C were found to have delayed the swelling and rupture of RBCs as compared with RBCs at 4°C and 37°C. However, 0.5% urea was seen to fasten the rupture of the RBCs at 4°C and 20°C as compared to that at 37°C where it was significantly delayed.
Key words: Hemolysis, Membrane permeability, RBCs
3. Journal of Cell and Tissue Research 15(2): 4949- 4955 (2015)
Expression of insulin receptors in insulin target organs in streptozotocin diabetic model: An immunohistochemical study
Kavitha Rani, B., Sugunarao., Byregowda, S. M., Satyanarayana, M. L., Narayanaswamy, H. D., Purushotham, K. M., and Gurikar, R. Y.
Department of Veterinary Pathology, Veterinary College, Karnataka Veterinary Animal and Fisheries Sciences University (KVAFSU), Shimoga 577204. E. mail:
Abstract: Insulin receptors are transmembrane proteins involved in insulin regulated glucose uptake signaling pathway, highly expressed in target tissues like liver, adipose tissue, skeletal muscle and also in several other types of mammalian cells. Several studies have documented a reduced expression of insulin receptors in insulin resistance and diabetes. In the present study, clinical diabetes was induced experimentally with streptozotocin in a rodent model, and maintained for 45 days along with non diabetic rats. Tissues like liver, heart, skeletal muscle and adipose tissues analyzed by immunohistochemical method using anti-insulin receptor α  antibody, showed expression of insulin receptors indicated by an intense brown coloration. It was noted that immunopositivity was observed in two locations viz., the cytoplasm and the plasma membrane of the target tissue cells. In the present study, a reduced expression of insulin receptors among the diabetic rats, when compared to non diabetic rats has been demonstrated by immunohistochemistry in liver, heart, skeletal muscles and adipose tissue, potentiating the fact that there is indeed a reduction in expression of insulin receptors in diabetes mellitus.
Key words: Insulin receptors, Diabetes, Immunohistochemistry
4. Journal of Cell and Tissue Research 15(2): 4957- 4965 (2015)
Evaluation of ROS induced genetic aberration in benzene intoxicated rat bone marrow
Roy, H. and Pillai, A.
Department of Zoology, Faculty of Science, The M.S. University of Baroda, Vadodara, Gujarat,
 E. mail:, Cell: 09924428489
Abstract: Here we aimed to observe the dose dependent genotoxic potential of benzene which may trigger the leukomogenic activity in bone marrow of rat. We used Wistar rat as a model and benzene was administrated orally at the doses of 50mg/ml/ day and 100mg/ml / day for 90 days which exhibited with definite signs of genotoxicity which may initiate carcinogenesis. Cellular stress of bone marrow was evaluated through various ROS parameter which was resulted into significant decrease in the level of antioxidative enzyme activity. There was significantly increased frequency of micronuclei in bone marrow of rat especially in polychromatic erythrocyte and the ratio of polychromatic and normochromatic erythrocyte was depleted at the significant level. Structural chromosomal aberration was observed significantly higher at 100mg/ml exposure of rat. The extent of DNA damage was evaluated through comet assay. A significant increase in the tail length of comet was recorded in the benzene exposed rats. Furthermore, the analysis of DNA ladder assay and cell morphology study of bone marrow cell was revealed the apparent hike of apoptotic cell death due to benzene intoxication. Present finding indicate that benzene exposure develops cancer pathogenesis in rat bone marrow by modulating the oxidative stress parameters and apoptosis pathway.
Key words: Benzene, Oxidative stress, Genotoxicity
5. Journal of Cell and Tissue Research 15(2): 4967- 4974 (2015)

Effect of respiratory disease on the bronchoalveolar lavage fluid cytology and bacteriology in sheep
Zaman, R. and Malik, H. U.
Division of Clinical Veterinary Medicine, Ethics and Jurisprudence, Faculty of Veterinary Sciences & Animal Husbandry, Sher-e-Kashmir University of Agricultural Sciences and Technology, Shuhama, Alusteng,Srinagar, Kashmir 190 006. E. mail:
Abstract: A study was undertaken to determine the effect of respiratory disease in sheep on the Bronchoalveolar lavage (BAL) cytology and bacteriology. Thirty two male housed sheep were subjected to fibreoptic bronchoscopy and BAL was collected. The BAL fluid of the affected sheep contained a significantly higher percentage of neutrophils (18.50 ±1.37 %) and lymphocytes (27.75 ±3.17 %) and a lower percentage of alveolar macrophages (39.25 ±5.15 %) than the group of healthy sheep (10.75 ±2.01, 14 ±2.73 and 62 ±6.60 % respectively). A low macrophage-to-neutrophil ratio was observed in sheep showing symptoms of respiratory diseases. Staphylococcus aureus was the most common bacteria isolated from BAL (41.67 %) of affected sheep. Other bacteria that were isolated included Klebsilla spp., Escherichia coli, Streptococcus spp and Mannheimia haemolytica. The bacteria were found to be sensitive to cephotaxime (84.21 %), gentamicin (78.95 %), enrofloxacin (78.95 %), chloramphenicol (68.42 %) and oxytetracycline (47.37 %) and resistant to streptomycin, ampicillin and cloxacillin. These results may allow BAL to assume a more prominent role in diagnosis and management of respiratory disease of sheep in the near future.
Key words: Sheep, Bronchoalveolar lavage, Fibreoptic bronchoscopy
6. Journal of Cell and Tissue Research 15(2): 4975- 4982 (2015)
Comparative pathogenesis of two spodopteralittoralis nucleopolyhedro virus isolates against
Culex pipiens larvae
Mahmoud, D. M. and Salem, D.A.M.
Department of Entomology, Faculty of Science, Ain Shams University, Abbassya, Cairo, Egypt
E. mail:,
Abstract: Viruses represent an important class of pathogens, and the most common viruses of mosquitoes are baculoviruses, which is considered as one of the most promising bioinsecticides. We estimated LC50 values of two local nucleopolyhedrovirus isolates from Spodopteralittoralis larvae (SLNPV) from two localities in Egypt; Giza and Cairo governorates; and their effect on the midgut region of third larval instar of Culex pipiens larvae after 6, 24 and 48 hours. LC50 values for Giza SlNPV and Cairo SlNPV were (1.7 X 10PIB/ml and 2.2 X 1010 PIB/ml.) respectively. In this investigation, an ultrastructure study, using semi-thin sections and electron micrographs was carried out to demonstrate the effect of the two isolates Giza SlNPV and Cairo SlNPV on the third larval instar of Cx. pipens. Treatment with Cairo SlNPV showed changes in the epithelial cells which returned back to its normal shape. Data reveals that Giza SlNPV was more efficient for control Culex pipiens larvae than Cairo SlNPV, and it can cause death to larval stage at 48h post infection with dramatic changes in the midgut layer, which makes this baculovirus as effective as insecticides, but in a safer form, and can be successfully included in the integrated pest management.
Key words: Baculoviruses, Nucleopolyhedrovirus, Culex pipiens larvae
7. Journal of Cell and Tissue Research 15(2): 4983- 4988 (2015)
Effect of diatomaceous earth on performance, internal organs and biochemical alterations
in T-2 toxicosis of broiler chickens
Shivashankar, B.P.,Narayanaswamy, H.D., Satyanarayana,  M.L., Rao, S., Rathnamma, D., Muniyellappa,
H.K. and Sridhar, N.B.
Department of Veterinary Pathology, Veterinary College, Bangalore 560024. 
E. mail:
Abstract: The efficacy of Diatomaceous Earth (DE) was evaluated in broiler chickens by adding 400 and 800 mg/kg in the feed and T-2 toxin was fed at 0.5 ppm and 1 ppm in the diet for 35 days of age. The body weight, FCR, relative organ weights and serum biochemical alterations were studied. A total of three hundred and sixty newly hatched unsexed day-old healthy commercial broiler chicks were randomly divided into nine groups consisting of control and treatment groups. Feeding of T-2 toxin resulted in significantly (P<0.05) reduced body weight and higher feed conversion ratio at the end of the trial. T-2 toxin at 1 ppm showed significantly (P<0.05) increased relative weights of liver and kidney and decreased relative weights of thymus and bursa of Fabricius compared to control. Whereas, significantly (P<0.05) higher kidney weight and decreased weight of thymus was noticed at 0.5 ppm of T-2 toxin in the diet. The serum biochemical alterations in T-2 toxin fed groups included increased ALT, AST and creatinine values and decreased total protein, albumin, triglycerides and cholesterol levels. Birds with dietary supplementation of DE showed significantly (P<0.05) higher body weight and lower feed conversion ratio and increased serum total protein and albumin values as compared to only T-2 toxin fed groups. However, DE supplemented birds did not differ significantly in relative organ weights of liver, kidney, thymus, spleen and bursa of Fabricius and serum biochemical values of ALT, AST, triglycerides, cholesterol and creatinine when compared to birds fed T-2 toxin alone. The study indicated that the addition of DE as an adsorbent in the diet was partially helpful in reducing the negative effects of T-2 toxin in broiler chickens.
Key words: T-2 toxin, Diatomaceous earth, Broiler chicken
8. Journal of Cell and Tissue Research 15(2): 4989- 4993 (2015)
Toxic effects of tefluthrin and aluminium on haematological parameters in wistar rats with special reference to ameliorative effect of alpha lipoic acid
 Mir, H.A., Sultana, M., Dar, A.M., Raina, R., Chirag, S., and Ahmed, A.
Department of Veterinary Pharmacology and Toxicology, IIVER, Rohtak, Haryana 124001
 E. mail:,
Abstract: The present study was aimed to investigate the protective role of alpha lipoic acid on various altered values of hematological parameters produced by sub lethal doses of tefluthrin and aluminium alone and in combination for a period of 28 days. Forty eight wistar rats of either sex weredivided into eight groups with six animals in each group. Group I served as control and were orally administered with corn oil. Groups II and III orally received tefluthrin and aluminium @ 1.1mg/Kg and 34mg/Kg respectively. Group IV animals were orally intoxicated both with tefluthrin @ 1.1mg/Kg and aluminium @ 34mg/Kg. Groups V received alpha lipoic acid @ 30mg/Kg where as groups VI, VIII and VIII respectively received tefluthrin, aluminium and combination thereof along with alpha lipoic acid. Tefluthrin and/or aluminium treatment produced a significant (P<0.05) decrease in the hematological parameters including hemoglobin (Hb), total erythrocyte count (TEC) and packed cell volume (PCV). Total leukocyte count (TEC) increased significantly (P<0.05) in all treated groups as compared to control. Groups treated with both tefluthrin and aluminium produced significant (P<0.05) changes in Hb and MCH as compared to groups treated with tefluthrin or aluminium alone. Pretreatment with alpha lipoic acid resisted the changes in hematological parameters produced by tefluthrin and/or aluminium.
Key words: Lipoic acid, Aluminium toxicity, Tefluthrin toxicity
9. Journal of Cell and Tissue Research 15(2): 4995- 4998 (2015)
Prenatal and postnatal changes in the biliary bile acid composition of Gaddi sheep
 Razvi, R.,Rajput, R.and Sharma, R.
Division of  Veterinary Anatomy, Himachal Pradesh Krishi Vishvvidlaya, Palampur, 176 061, Himachal Pradesh.
 E. mail:,
Abstract: The biliary bile was collected from different prenatal and postnatal age groups of Gaddi sheep. Prenatal age groups included; 1-60 days (group I), 61-120days (group-II) and 121 –full term (group-III), while the postnatal age groups included; neonatal (below 1 year; group-IV), young (2-3 years; group-V) and adult (above 3 years of age, group-VI). Biliary bile was collected from nine animals in each group. Thin layer chromatography was done and retention factor (Rf) was recorded to observe variable composition of three bile acids: chenodeoxycholic acid (CDCA), cholic acids (CA) and deoxycholic acid (DCA). The Rf  value of CDCA was maximum (0.43) in group III and minimum (.055) in group VI. The proportion of cholic acid was lesser in prenatal groups as compared to postnatal groups and its maximum value was recorded in neonatal (group IV). The DCA- a secondary bile acid, its minimum Rf value (0.023) was observed in neonatal group (group IV) and highest (0.207) in adult Gaddi sheep (group VI). The transition of these bile acids from prenatal to postnatal age groups, indicated the limited placental transfer of bile, development of an anaerobic cecum and change of bile acid proportion with age.
Key words: Bile acids, Gaddi sheep
10.    Journal of Cell and Tissue Research 15(2): 4999- 5011 (2015)
Modulating effect of the aerial parts of pumpkin methanol extract in liver and pancreatic islets on experimental diabetic rat: histological, biochemical and immunochemical studies
Ahmed, F.E.,Selim, A.O., Selim, S.A. and Mohamed, A.M.
Department of Histology & Cell Biology, Faculty of Medicine, Zagazig University, Zagazig, Egypt. 
 E. mail:
Abstract: Nowadays, the herbal is considered a novel therapeutic approaches in the treatment of diabetes. Pumpkin extract is the most recommended one is seen to help in regulation of blood sugar. It is prescribed widely even when its biologically active compounds are still unknown. This work is designed to evaluate the protective role of methanol extract of aerial part of Cucurbita maxima (MECM) on experimental streptozotocin (STZ) induced pancreatic islets and hepatic injury. Forty male rats were divided into 4 groups: I: control; II: STZ induced diabetes; III: diabetic rats received daily 5 mg/kg of Gilbenclamide (GLB) and IV: diabetic rats received daily 200 mg/kg of MECM. At the end of experiment (30 days), the rats were sacrificed and blood samples were collected for different biochemical analysis. Livers and pancreases were removed and prepared for histopathological examination. It was found that the liver of diabetic rats revealed histological and chemical changes versus the control. The administration of MECM decreased glucose level as well as the lipid profile to/towards near the normal. On the other hand, the normal histological features were nearly resumed in the liver and pancreas which further evidenced the hepatoprotective activity of MECM. The present study proved a lessening effect of MECM on the diabetic liver. It is advisable to widen the scale of its use, for patients at high risk of diabetes mellitus in a trial to alleviate the hepatic hazards.
Key words: Streptozotocin, Pumpkin, Liver and pancreas, Rat, Histology.
11.    Journal of Cell and Tissue Research 15(2): 5013- 5018 (2015)
Assessment of biochemical diversity in green gram genotypes through isozymes
 Kaur, G., Joshi, A., Jain, D., Rajamani, G., Vyas, D. and Jain, R.K.
Department of Molecular Biology and Biotechnology, Rajasthan College of Agriculture, Maharana Pratap University of Agriculture and Technology, Udaipur-313001, Rajasthan. E. mail:,
Abstract: Amongst the pulses, greengram is an crop of excellent source of high (25%) quality dietary protein in the vegetarian diet. In the present investigation, isozymic patterns and protein profiling were studied in 23 genotypes of Vigna radiata L. peroxidase (PER), Esterase (EST), and superoxide dismutase (SOD) isozymes were screened by native polyacrylamide gel electrophoresis (PAGE) technique to study the genetic divergence. A total of 10 alleles were generated by the three isozymes. Isozymes of SOD and EST each exhibited a maximum of four activity zones followed by PER that showed only two activity zones. Isozymes of EST and SOD showed 50 and 25 per cent polymorphism with all alleles. Jaccard’s Similarity Coefficient values lay between 0.73 to1.00 that divided all the genotypes into two main clusters. Genetic divergence and distance derived from isozyme analysis was found low due to the small number of polymorphic alleles. PIC values ranged from 0.07 to 0.16 with an average of 0.115. The total soluble protein content was found maximum in genotype HUM-12 (16.8 mg/g) whereas minimum was observed in genotype ML-729(8.8mg/g). Variation in the isozymic profiles amongst greengram genotypes may be an indication of genetic variation that could be exploited in breeding programmes.
Key words: Green gram,Isozymes, Genetic divergence
12.  Journal of Cell and Tissue Research 15(2): 5019- 5024 (2015)
Incidences of nuclear anomalies in human in relation to quantity and duration of gutkha consumption
 Jaggi, S. and Yadav, A.S.
Human genetics Laboratory, Department of Zoology, Kurukshetra University, Kurukshetra 136119, Haryana.
E. mail: ,
Abstract: The gutkha consumption is very popular among a section of society, especially in socio-economically backward and illiterate. Chewing of gutkha is believed to be associated with oral and hypopharyngeal cancers. The present study was aimed to evaluate the genetic damage in buccal exfoliated epithelial cells of gutkha consumers using micronucleus assay. The study comprises 100 individuals of which 50 individuals were gutkha consumers (25 males and 25 females) and 50 were control subjects (25 males and 25 females). A significantly (P<0.001) high frequency of micronuclei and other nuclear anomalies were observed among gutkha consumers as compared to control subjects. Significant (P<0.01) positive correlation was observed for micronuclei frequency in relation to number of pouches of gutkha consumed per day and duration of gutkha consumption (years). The results of the present study suggest that gutkha is a genotoxic agent and induces genetic damage.
Key words: Gutkha, Nuclear anomalies, Human buccal epithelial cells
13.    Journal of Cell and Tissue Research 15(2): 5025- 5028 (2015)
Comparative efficacy of various therapeutic protocols on haematological and biochemical profiles of bitches affected with Pyometra
Ahmed, A., Dar, A.M., Bhat, A.A., Chowdhary, A.R.S., Jena, B., Pagrut, K.N., Sharma, N. and Bhainsare, P.
Department of Teaching Veterinary Clinical Complex,  M. J. F. College of Veterinary and Animal Sciences, Rajasthan University of Veterinary and Animal Sciences, (Rajasthan).
Abstract:: Pyometra is a potentially life threatening condition which requires an early diagnosis to conserve the life of the animal. The analysis of various parameters helps in the assessment of the clinical status of the bitches and in prediction of the prognosis. This study was undertaken on canine pyometra and consisted of 20 bitches of different breeds with known breeding history and open type of pyometra. The haematological and biochemical parameters were evaluated before and at the end of treatment. Haematological values showed decrease in Hb, PCV, TEC and lymphocyte count in bitches affected with pyometra. Biochemical studies revealed elevated monocytes, neutrophils, BUN, globulin and creatinine level in affected bitches. The clinical cases were divided randomly into four different groups with each group consisting of five bitches. One group of bitches was kept as untreated control group in which only supportive therapies were given. The bitches in the other three groups were treated using antibiotics with supportive therapies, natural PGF2α along with antibiotics and the fourth group was treated with dopamine agonist prolactin inhibiting drug, i.e., cabergoline along with combination of a synthetic PGF2α analogue. Treatment of canine pyometra by the use of different drugs was found to be successful. Treatment of canine pyometra using a combination natural PGF2α along with antibiotics was found to be the most effective method among the three therapeutic protocols used in the present study.
Key words: Pyometra, Bitches
14.    Journal of Cell and Tissue Research 15(2): 5029- 5036 (2015)
Biochemical alteration in tomato varieties resistant and susceptible to Fusarium oxysporum
F. sp. lycopersici
Solanki, V.H., Patel, R.M. and Patel, K.G.
Department of Plant Molecular Biology and Biotechnology, N. M. College of Agriculture Navsari Agricultural University, Navsari 396450 (Gujarat). E. mail:,
Abstract:: In present contribution Fusarium oxysporum f. sp. lycopersici, causing Fusarium wilt, induced changes in biochemical composition of tomato varieties viz., resistant (Junagadh Tomato-3 and TBK-01113), moderately resistant (Gujarat Tomato-2 and Anand Tomato-3) and susceptible (Pusa Ruby and Junagadh Ruby) was studied. Study shows alteration in total phenol content and enzymes viz. catalase, peroxidase, superoxide dismutase, polyphenol oxidase, phenylalanine ammonia lyase and β-1,3-glucanase (B13g) activity in different varieties of tomato at pre-infection (one day before infection i.e. 20 days after sowing) and post-infection stages (7 days after infection (DAI) i.e. 28 DAS, 14 days after infection (DAI) i.e. 35 DAS, 21 days after infection (DAI) i.e. 42 DAS). Results reveal that inoculated resistant varieties had significantly higher total phenol content, catalase, PAL, PPO and (B13g) activity on 7th days after infection and their concentration progressively reduced up to 21st day. The highest peroxidase activity was found on the 14th days after infection. Whereas, highest SOD activity was observed in susceptible varieties of tomato at 21st day.
Key words: Fusarium oxysporum, Fusarium wilt, Tomato varieties
15.  Journal of Cell and Tissue Research 15(2): 5037- 5041 (2015)
Effect of NACL induced salt stress on proline, mda and antioxidant mechanism in rice (Oryza sativa L.)
Srivashtav, V.,Rajkumar, Jha, S.and Singh, D.
Department of Plant Molecular Biology and Biotechnology, Navsari Agricultural University, Navsari, Gujarat. E. mail:, Cell: 7600059128
Abstract:: The aim of this study was to examine possible alterations in the metabolism of rice plants (Oryza sativa L.) that were exposed to high salt concentrations. NAUR1 seedlings were treated with Hoagland’s solution containing 25, 50, 100 and 200 mM NaCl and maintained for 14 days in these conditions. After 14 days there was a gradual increase in the levels of lipid peroxidation, proline and catalase (CAT); and reduced activity of antioxidant enzymes viz., superoxide dismutase (SOD), and guaiacol peroxidase (GPOX) with increasing salt concentrations in the Hoagland’s solution. Glutathione reductase (GR) activity increased at 25 mM but decreased at 50, 100 and 200 mM salt concentrations. These results suggested that proline may have protective effects against protein degradation. Moreover, although antioxidant enzymes such as SOD and GPOX shown to possess low levels of activity, a large proportion of the hydrogen peroxide that produced is preferentially directed towards lipid peroxidation. High catalase and lower guaiacol peroxidase activities may be regarded as potentially useful biochemical indicators for selection of salt tolerant rice cultivars and targets for improvement through transgenic approaches.
Key words: Salt stress, Oryza sativa
16.  Journal of Cell and Tissue Research 15(2): 5043- 5046 (2015)
Genetic variability within and between Cucumis species through SSR markers
 Rahul, K., Imran, D., Sudarshan, Y., Sharma, R., Singh, G. and Rizwan, M.
Plant Biotechnology Centre, Swami Keshwanand Rajasthan Agricultural University, Bikaner Rajasthan .
E. mail:,
Abstract:: Genetic diversity in 30 of Cucumis melo and 10 each of C. melo var. callosus and momordica collected across Rajasthan and nearby area were evaluated through SSR marker. Out of 11 SSR primers tested, seven primers amplified and out of seven,   three SSR markers (SSR 00001> 12682> 16301) showed variability in C. melo while SSR 19343 displayed variability in C. melo var. callosus.. The similarity in annealing temperature of each of the seven SSR markers for any two of the three groups (C. melo, C. melo var. momordica and C. melo var. callosus) indicated a common genetic pool among these accessions. A small set of four SSR markers viz., 00001, 12682, 16301 and 19343 were sufficient differentiate intra and inter group accessions of C. melo and C. melo var. callosus.
Key words: Genetic diversity, Cucumis melo
17.  Journal of Cell and Tissue Research 15(2): 5047- 5052 (2015)
Association of sugar and phenol content with fusarium wilt resistance in pigeonpea
 Patil, V.R., Patel, R.M. and Parekh, V.B.  
Department of Plant Molecular Biology and Biotechnology, NMCA, Navsari 396 450, Gujarat.
E. mail:,
Abstract:: The study was undertaken using wilt resistant pigeonpea cultivars: Vaishali, Maruti and susceptible cultivars: T15-15, Bahar. Total soluble sugars and total phenols were estimated using 15 days old seedlings infected with fusarium wilt pathogen Fusarium udum and control seedlings. Lower leaf samples were collected at all three different stages (0 days, 4 and 8 DAI), while lateral roots were collected only at two stages (0 days and 8 DAI). In resistant cultivars of pigeonpea, sugar content in leaves as well as root tissues was drastically reduced whereas, susceptible plants had pronounced induction as diseases progressed compared to pre-infection. In resistant cultivars total phenols content in leaf was increased (48.17 %) over control in cultivar Maruti at 4 DAI and then, it was reduced (2.67 %) at 8 DAI, cultivar Vaishali also showed same trend. In susceptible cultivars, total phenol was decreased at 4 DAI only in cultivar T15-15, however, it was registered higher at 8 DAI over the control in both susceptible cultivars T15-15 and Bahar. In case of root, total phenol was reduced in both resistant cultivars. The higher level of sugar during fusarium wilt disease was associated with wilt susceptible pigeonpea cultivar and higher level of phenol at earlier stages associated with wilt resistant cultivars.
Key words: Fusarium wilt, Pigeonpea
18.    Journal of Cell and Tissue Research 15(2): 5053- 5057 (2015)
The viability of yoghurt probiotic culture in microencapsulated iron fortified yoghurt
Subash, R., Elango, A., Pandiyan, C., Karthikeyan, N., Kumaresan, G. and Raghunath, B.V.
Department of Dairy Science, Veterinary College and Research Institute, Namakkal, 637 002, Tamil Nadu.
E mail:,
Abstract:: A study was designed to develop microencapsulated whey protein-chelated iron (Fe-wp) using ferrous sulphate as the iron source that could be used in the development of iron fortified yoghurt. Influence of iron on survival of yoghurt culture, TBA values of yoghurt and sensory properties of yoghurt were tested by control, free iron and encapsulated iron fortification. Statistically no significant (P>0.05) difference was noticed in count of Lactobacillus delbrueckii ssp. bulgaricus and Streptococcus salivarius ssp. thermophilus between control and different iron fortified yoghurt treatments on 0, 7, 14 and 21 days. During storage period, the count of Lactobacillus delbrueckii ssp. bulgaricus and Streptococcus salivarius ssp. thermophilus significantly (P<0.05) decreased both in control and as well as in iron fortified yoghurt and thus the fortified iron did not affect the viability of yoghurt bacteria. The TBA values of unencapsulated iron fortified yoghurt was significantly (P<0.05) higher when compared to control and encapsulated iron fortified yoghurt. Significant (P<0.05) difference was observed in oxidized flavour at 0, 7, 14 and 21st day of storage between control and different treatments of yoghurt. In addition, significant (P<0.05) difference was observed in overall preference at 0, 7, 14 and 21st day of storage between control and different treatments of yoghurt and between different storage periods. The present study demonstrated that microencapsulated whey protein chelated iron can be added up to a level of 80 mg per litre of yoghurt using ferrous sulphate without affecting the viability of yoghurt probiotic culture.      
Key words:- Yoghurt, Microencapsulation, Whey protein chelated iron
19.  Journal of Cell and Tissue Research 15(2): 5059- 5061 (2015)
Isolation and detection of Escherichia coli from diarrheic calves alongwith biochemical and hematologic parameters
 Bashir, S., Ashraf, I., Dar, A.M., Majid, A.,Bhat, A.A., Farooq, J.
Division of Epidemiology and Veterinary Preventive Medicine, DUVASU, Mathura, UP;
E. mail:,
Abstract:: This diagnostic study was conducted to isolate Escherichia coli from calves affected with neonatal diarrhea (Colibacillosis) and determine the changes in hematologic and biochemical parameters. E. coli was isolated and identified from fecal samples by cultural and biochemical characteristics. 16s rRNA gene of E. coli was detected by PCR. 16 of 25 samples were positive for E. coli isolation. There were marked alterations in hematological findings with elevated PCV, TEC, and TLC. Diarrheic calves were neutrophilic but lymphopenic. Biochemical analysis revealed lowered sodium level, and increased chloride and potassium levels. This study suggests colibacillosis an important cause of neonatal diarrhea in calves.
   Key words: Diarrheic calves, E. coli
20.    Journal of Cell and Tissue Research 15(2): 5063- 5066 (2015)
Isolation of infectious bovine rhinotracheitis virus in cell culture: (A case study)
 Bashir, S., Dar, A.M., Bhat, A.A.,Bukhari, S.,Hussain, T., Yadav, K.S., and Ahmed, A.
Division of Epidemiology and Veterinary Preventive Medicine, DUVASU, Mathura, UP
E. mail:,
Abstract:: Infectious bovine rhinotracheitis is an important disease of cattle with serious economic implications. The etiology is bovine herpes virus type-1 which leads to clinical to latent infections and abortions. In this study, an aborted bovine fetus was suspected of IBR infection. The virus was grown in cell line (MDBK) and cytopathic effect observed. CPE began at 12 hr post infection of cells and was quite evident 96 hr post infection with complete degeneration of cells which were floating in the medium. Cell culture cultivation is an important method for isolation and detection of this virus. 
Key words: Bovine rhinotracheitis, Bovine herpes virus type 1
21.  Journal of Cell and Tissue Research 15(2): 5067- 5071 (2015)
Efficacy of growth hormone for callus induction in sesame (Sesamum indicum L.)
Pusadkar, P.P., Kokiladevi, E., Aishwarya, V., Gnanam, R., Sudhakar, D. and Balasubramanian, P.
Department of Plant Biotechnology, Centre for Plant Molecular Biology and Biotechnology, Tamil Nadu Agricultural University, Coimbatore 641003, Tamilnadu. E. mail:
Abstract: An efficient procedure was developed for continuous callus induction from the hypocotyl explants of two sesame cultivars. Explant and growth regulators used were shown to be critical factors for callus induction. The potential for in vitro response of cotyledonary and hypocotyl explants of sesame (S. indicum L) cv. TMV 7 and G1 were studied on MS media supplemented with different concentrations of growth hormones such as auxins (NAA, 2,4-D, IAA) cytokinins (BAP, KIN, TDZ) along with ABA and AgNO3 for callus induction. Explants were cultured on MS medium supplemented with different concentrations (1-5 mg/l) of individual growth hormones (BAP, KIN, NAA, 2, 4-D, TDZ, IAA). Callus formation was not observed when individual hormones at different concentrations were used. The combinations of growth regulators (BAP, KIN, NAA, 2, 4-D, TDZ, IAA) along with ABA and AgNO3 were used for callus induction using cotyledonary and hypocotyl explants. Only hypocotyl explants showed the best response for callus induction. MS medium containing NAA (0.5 mg/l) and TDZ (1 mg/l) produced yellowish green friable calli with highest callus induction efficiency of 93.3% in cv. TMV 7 and 90% in G1. The morphology of the calli varied with the different combination of growth regulator used from greenish yellow to pale white from friable, compact to watery calli.
Key words: Sesamum indicum L, Hypocotyl explants, Callus
22.  Journal of Cell and Tissue Research 15(2): 5073- 5077 (2015)
Molecular detection and cardiopulmonary pathological studies of porcine pasteurellosis in pig population of Chhattisgarh
Tigga, M., Ghosh, R.C., Malik, P.,Choudhary, B.K. and Mishra, C.S.
Department of Veterinary Pathology, College of Veterinary Science and A.H., Kamdhenu Vishwavidyalaya, Durg, Anjora, Chhattisgarh; E. mail:,
Abstract: Outbreaks of swine pasteurellosis were observed in descript as well as non-descript pigs of Chhattisgarh, India. It is an economically important infectious disease of pigs, has brought great economic losses to the pig farmers by high rates of morbidity and mortality. Affected animals exhibited pyrexia, dullness, staggering gait, anorexia, serous nasal discharge and dyspnoea.Affected pigs revealed 66% mortality and 100% case fatality rates in pre-weaned piglets. The article describes the molecular characterization and cardiopulmonary pathological studies on swine pasterurellosis. Diagnosis of disease was confirmed by PCR on Pasteurella multocida isolated from clinical samples. Lungs, the predominantly affected organ were severely congested with varying degrees of consolidation and with a marked thickening of the interalveolar septa, pleura and rubbery consistency of lungs. Heart was severely congested and there were presence of petechial as well as haemorrhagic streaks and necrotic foci visible upon removal of pericardium. Histopathological examination showed haemorrhages and necrosis of myocardium, acute fibrinous pneumonia characterized by serofibrinous exudation and infiltration with polymorphonuclear cells, macrophages and erythrocytes.
Key words: Swine pasteurellosis, Pneumonia, Pasteurella multocida
23.  Journal of Cell and Tissue Research 15(2): 5079- 5084 (2015)
The impact of feeding propylene glycol, bypass fat and bypass protein on progesterone concentration
 in postpartum dairy cattle
Shankare Gowda, A.J.,Devaraj, M., Krishnaswamy, A., Ranganath, L.,Ravindra, J.P. and Gupta, P.S.P.
Department of Animal Reproduction, Gynaecology & Obstetrics, Veterinary College, Vinoba nagar, Shimoga 577204; E. mail:,
Abstract: High yielding dairy cows cannot consume adequate nutrients in early lactation to support the level of milk yield. The peak milk production, at about 5-8 weeks postpartum occurs earlier than maximum feed consumption causing cows to be in negative energy balance and to mobilize fat from adipose tissue. Negative energy balance affects gonadotropin secretion, ovarian physiology and ovarian hormone production which in turn adversely affects the reproductive performance of cattle during postpartum period. The present study was under taken to evaluate the influence of propylene glycol, bypass fat and bypass protein on postpartum luteal activity as the interest was being grown among dairy farmers, veterinarians and advisers. A total of twenty eight postpartum dairy cows were randomly allotted to four groups with seven animals in each group. The Group-I animals served as control without being fed any feed additive. Group-II animals received propylene glycol once daily as a drench at the rate of 300 ml per head while Group-III and Group-IV animals were fed 100 and 200 gms of commercially available bypass fat and bypass protein respectively. In all the groups, the mean serum progesterone concentration was at basal level on day 7 and 14 postpartum and subsequently its level increased with clear higher level from day 21 onwards. The difference in mean progesterone levels at different stages of the study was significant (Pd”0.05) and the increase seen was almost linear with highest levels on day 63 postpartum. Although, the stage variation between groups was significant, the higher or lower values recorded at different stages was not consistent with a given group.
Key words: Propylene glycol, Bypass protein, Bypass fat, Progesterone, Dairy cattle
24.  Journal of Cell and Tissue Research 15(2): 5085- 5087 (2015)
Incidence of abortion in relation to its parity and season in Holstein Friesian upgraded dairy cows
Shankare Gowda, A.J., Honnappa, T.G., Nagaraju, C.S., Devaraj, M.and Krishnaswamy, A.
Department of Animal Reproduction, Gynaecology & Obstetrics, Veterinary College, Vinoba nagar, Shimoga-577204. E. mail:,
Abstract: Abortion is major threat to dairy industry causing fetal loss and prolonged uterine pathology followed by infertility and sterility. Data on HF upgraded cows revealed that overall incidence of abortion to be 7.53%. Parity and stage of gestation found to have no influence on the incidence of abortion while season showed a significant effect on it. High incidence of abortion was recorded during northeast (2.25%) and southeast (2.07%) monsoon seasons. Since exact cause for this incidence of abortion is unknown, further studies are needed to ascertain the cause.
Key words: Abortion, Holstein Friesian upgraded Cows.
25.    Journal of Cell and Tissue Research 15(2): 5089- 5094 (2015)
Assessment of biochemical diversity through isozyme analysis in black gram (Vigna mungo l.) genotypes
 Vyas, D., Joshi, A., Jain, D., Rajamani, G. and Kaur, G.
Department of Molecular Biology and Biotechnology, Rajasthan College of Agriculture, Maharana
Pratap University of Agriculture and Technology, Udaipur-313001, (Rajasthan). E. mail:  
Abstract: Vigna mungo L. is considered to be one of the important pulse crop of India. In the present study 22 genotypes of V. mungo L. were collected from different geographical locations of India. Total soluble protein was estimated and three isozymes viz., esterase (EST), peroxidase (POX) and superoxide dismutase (SOD) were screened by native polyacrylamide gel electrophoresis technique to study the variation in isozymatic patterns. The total soluble protein was ranged from UH-86-5 (9 mg/g) to RUG-8 (17.4 mg/g) respectively. Based on isozymic profile a total of 14 alleles were generated. Isozymes of SOD exhibited a maximum number of activity zones i.e., 7 followed by EST (4) and POX (3). Average polymorphism was 53.96 %. PIC values ranged from 0.08 to 0.30 with an average of 0.23. Jaccards similarity coefficient values ranged from 0.76 to 1.00, revealed that the genetic distance derived from isozyme analysis was very low due to small number of polymorphic bands. Cluster tree analysis carried out by UPGMA method divided all genotypes into two clusters at a similarity coefficient value of 0.80. Isozymes of esterase showed 100% polymorphism. The esterase banding pattern could be used as a biochemical marker for the identification of species in V. mungo L.

Key words: Vigna mungo L., Genetic diversity, Isozymes
26.    Journal of Cell and Tissue Research 15(2): 5095- 5100 (2015)
Adhesion properties of riboflavin producing Lactobacillus mucosae isolated from human faeces
 Thakur, K.,Gosh, S.,Kadam. J.  and Tomar, S.K.
National Collection of Dairy Cultures, Dairy Microbiology Division, National Dairy Research Institute, Karnal 132001, Haryana. E. mail:,
Abstract: In the process of selecting new riboflavin producing isolates, several Lactobacilli were isolated from human faeces. Among them one strain identified as Lactobacillus mucosae KTF was characterized on the basis of 16srDNA. In this study, we have assessed the adhesion property of Lactobacillus mucosae KTF based on its autoaggregation phenotype and presence of mub gene (mucin binding factor). The mucin degradation test was performed to ensure the safety of the culture. This riboflavin producing strain has displayed the presence of mubgene, appreciable in vitro adhesion on HT-29 cell lines investigated by % adhesion method, adhesion score method, giemsa stain method and Scanning electron microscopy and thus fulfilled the criteria to be identified as a putative probiotic microorganism. Evaluation by mucin degradation test showed the test strain is unlikely to degrade the mucin in plate assay method. The strong adhesion may be useful for competitive exclusion and displacement of pathogens as well as for functional foods preparations in the future. Further research is required to identify the other attributes related to adhesion and pathogen exclusion properties of this potential putative indigenous riboflavin producing strain.
Key words: Lactobacillus, Riboflavin, Human faeces
27.    Journal of Cell and Tissue Research 15(2): 5101- 5103 (2015)
Antibiotic sensitivity pattern of Aeromonas spp. isolated from food animals and immunocompromised cancer patients
 Tanuja Gowda, K.G.M., Zade, N.N., Chaudhari, S.P., Khan, W.A., Shinde S.V. and Kadagi, M.
Department of Veterinary Public Health, Nagpur Veterinary College, Maharashtra Animal and Fishery Science University, Seminary Hills, Nagpur 440006 (Maharashtra); E. mail:
Abstract: A total of 49 Aeromonas isolates obtained from food animals and immunocompromised human cancer patients from Nagpur, were investigated for antibiotic sensitivity pattern by disc diffusion method. Aeromonas spp. exhibited different level of antibiotic sensitivity based on the zone of inhibition observed around the antibiotic disc. Most of the isolates exhibited highest sensitivity towards ceftazidime, amikacin, ceftriaxone, cefotaxime and doxycycline hydrochloride. Drugs which showed significant resistance were ampicillin, amoxyclav and ampicillin/sulbactum. Among the isolates of Aeromonas spp. tested, Aeromonas hydrophila and Aeromonas sobria exhibited maximum sensitivity towards ceftriaxone whereas, Aeromonas caviae showed highest sensitivity towards cefotaxime. These results demonstrated the presence of wide spread resistant Aeromonas spp. in the region.
 Key words: Aeromonas spp. Antibiotic sensitivity, Cancer patients
28.    Journal of Cell and Tissue Research 15(2): 5105- 5110 (2015)
Acidosis reduced the sensitivity of l-type Ca2+ channels in goat superior mesenteric artery
 Mohanty, I. and Parija, S.C.
Department of Pharmacology and Toxicology, College of Veterinary Science and Animal husbandry, Orissa University of Agriculture Technology, Bhubaneswar 751003 E. mail:,
Abstract: In acidosis the relative effectiveness of calcium channel blockers (CCBs) in normalizing the hypertension could be affected. The sensitivity of L-type Ca2+ -channels to CCBs in goat superior mesenteric arterial rings under acidosis was examined using standard isometric contraction study. KCl(85mM) or NA(10µM) induced sustained contraction was decreased from pH 7.4   6.8  6.0 in the ratio of 1.00: 0.87: 0.79 (KCl) and 1.00: 0.84: 0.56 (NA), respectively. Vasotonic response induced by KCl and NA(100%) was inhibited by nifedipine (92.82±1.99%, 42.79±5.08%) at pH 7.4, (88.76±0.38%, 19.31±0.81%) at pH 6.8 and (77.34±2.49%, 14.75±0.80%) at pH 6.0. Amlopdipine inhibited vasotonic response of KCl and NA by (78.37±4.08%, 48.04±4.92%) at pH 7.4, (56.70±4.12%, 39.97±4.66%) at pH 6.8 and (57.39±4.03%, 17.08±3.75%) at pH 6.0. In conclusion, acidosis caused reduced sensitivity of L-type Ca2+ channels to CCBs in vascular bed. The clinical implication is that hypertensive patients when normotensive after treatment with CCBs if develops hyperacidosis could exhibit hypertension.
Key words: L-Type Calcium Channels, Goat mesenteric artery, Hypertension
29.    Journal of Cell and Tissue Research 15(2): 5111- 5115 (2015)
Elimination of bacterial contamination by using antibiotics in micropropagtion of banana
(Musa spp.) cv. Grand naine
 Sabale, S.N., Patil, D.M., Gokhale, N.B., Sawardekar, S.V. and Sawant, S.S.
Plant Biotechnology Centre, Dr. Balasaheb Sawant Konkan Krishi Vidyapeeth, Dapoli
Dist. Ratnagiri 415 712 (M.S.). E. mail:, 
Abstract: In banana, the major factor affecting the efficiency of micropropagation systems is the occurrence of bacterial contamination. Bacterial growth observed around the explants in the culture media results into destruction of large number of plantlets in the culture. Hence, the use of different antibiotics in medium during micropropagtion of banana (Musa spp.) Cv. Grand Naine were found to be an effective method to eliminate the bacterial contamination. Out of a four antibiotics tested in MS medium suggests that, the most suitable antibiotic concentration to be used in media during establishment and proliferation in banana genotype Grand Naine was 1000 ppm amoxicillin which showed maximum elimination of bacterial contamination at 30 DAI. The lowest incidence of bacterial contamination (11.667) and maximum explants survival (88.333) at 30 DAI was observed in the media combination MS + 5 mgl-1 BAP + 2 mgl-1 IAA + 1000 ppm amoxicillin. The maximum numbers of shoot initiation was observed in media combination MS + 5 mgl-1 BAP + 2 mgl-1 IAA + 1000 ppm amoxicillin while, the minimum number of days to shooting was recorded (24.75 days) in media combination MS + 5 mgl-1 BAP + 2 mgl-1 IAA + 1000 ppm cefotaxime. The media combination MS + 5 mgl-1 BAP + 2 mgl-1 IAA + 750 ppm amoxicillin produced highest number of shoots per explant i.e. 6.24 shoots per explants.
Key words: Banana, Bacterial contamination, Amoxicillin
30.    Journal of Cell and Tissue Research 15(2): 5117- 5124 (2015)
Analysis of Booroola fecundity (FecB) gene using PCR-RFLP with Haeiii and Xhoi restriction endonucleases in garole and Muzaffarnagri sheep
 Kumar, P., Singh, R.V., Chaudhari, M., Dubey, T. and Tomar, A.K.
Indian Veterinary Research Institute, Izatnagar 243122 (U.P.). E. mail:,
Abstract: Booroola fecundity (FecB) gene mutation is attributed to high prolificacy in Garole sheep. In present study to identify a RFLP marker of FecB gene of Garole sheep, PCR-RFLP method was used to study polymorphism of exons 8 (223 bp) and 10 (153 bp) of Booroola fecundity (FecB) gene. Specific exons were amplified from DNA extracted from the blood samples of 53 Garole sheep and 10 Muzafarnagari sheep. Digestion of amplified product was carried out with HaeIII and XhoI restriction enzymes. In both breeds of sheep HaeIII restriction site was present in all animals resulting in a single restriction pattern consisting of two fragments. HaeIII digestion of exon 8 yield fragments of 61 and 162 bp and XhoI digestion of exon 10 yield 18 bp and 135 bp fragments. All the animals studied showed similar restriction fragment pattern. These showed that, HaeIII and XhoI sites were not polymorphic in the Garole and Muzafarnagari sheep subpopulation in this study. Exon 8 and 10 were analyzed by DNA sequencing of the cloned products and a phylogenetic analysis was carried out to interpret the divergence among major species.
Key words: FecB gene, Garole, Muzzaffarnagri, PCR-RFLP
31.    Journal of Cell and Tissue Research 15(2): 5125- 5132 (2015)
PCR-RFLP based leptin gene polymorphism and its association with mastitis in Murrah buffalo
Anand, V. K., Sharma, V., Kumar, A., Verma, V.K., Rana, V.P., Khirbat, R. and Sangwan, M.L.
Department of Animal Biotechnology, Lala Lajpat Rai University of Veterinary and Animal Sciences,
 Hisar, Haryana, 125004. E. mail:,
Abstract: Due to the role of leptin gene in the immunity of animal, the present study was carried out to find the association of leptin gene with mastitis by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). Genomic DNA was isolated from the blood of randomly selected, lactating 40 Murrah buffaloes (including 20 mastitic animals) by phenol chloroform isoamyl alcohol method. PCR amplification of the partial intron2 (213 bp) and exon3 (331 bp) of leptin gene was carried out using self-designed primers and published primers, respectively. Resolution of digested, 213 bp PCR products of intron2 and 331 bp PCR product of exon3, with AluI and HphI; and HinfI and HaeIII restriction enzymes respectively, detected single genotype in both groups. The sequence of partial leptin gene (intron2) was found 100% identical to Bubalus bubalis, 98% identical to Bos taurus and 97% identical to Bos indicus, Capra hircus and Ovis aries leptin gene sequences. While the sequence of partial leptin gene (exon3) was found 100% identical to Bubalus bubalis, 98% identical to Bos taurus and Bos indicus, 94% identical to Capra hircus and 92% identical to Ovis aries leptin gene sequences. So we conclude that the partial region of leptin gene viz. intron2 and exon3 are monomorphic and could not be associated with the mastitis.
Key words:Leptin, PCR-RFLP, AluI, HphI, HinfI, HaeIII

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