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1. Journal of Cell and Tissue Research 14(2): 4181- 4192 (2014)
Histopathological effect of Spodoptera littoralis nucleopolyhedrosis virus (slnpv), Bacillus Thuringiensis kurstaki (btk), and their combination on Culex pipiens l. larvae (Culicidae: Diptera)
Abd El-Bar, M.M. and Mahmoud, D. M.
Department of Entomology, Faculty of Science, Ain Shams University, Abbassya, Cairo, Egypt.
E. mail: marah_elnaggar@yahoo.com
Abstract: The aim of the present work is to seek the histopathological alterations in Culex pipiens larvae midgut that may explain to some extent, the synergistic effect of SlNPV-Btk combination treatment which was noticed in our previous work.Third larval instar of Culex pipens were exposed to LC50 of SlNPV, Btk and SlNPV-Btk (1.3 X 103 PIB/ml, 2.5 X 105 spores/ml and 3.6 X 106 mixed polyhedra and spores/ml) respectively, and alterations in midgut region were recorded 6, 12 and 24 h post-infection. Data showed increasing damages of Cx. pipiens larval midgut epithelium for all three treatments but acceleration in midgut alterations was more evident in case of SlNPV-Btk combination sections. The SlNPV histopathological results revealed a serious damage on epithelial cells, Brush boarder, nuclei and basement membrane. The peritrophic membrane (pm) was not completely lysed untill 24h post-infection. Btk treatment of Cx. pipiens larvae promoted structural alteration changes in midgut and by 24h post infection, swelling and dislodging were observed in epithelial cells but no lyses. SlNPV-Btk combination had proven to be more effective against Cx. pipiens larval midgut cells than using either SlNPV or Btk solitary. Serious alterations were recorded by 6h post-treatment. Moreover, the abnormal aggregation of mitochondria, the bulbous-shaped microvilli, and the nuclear tubules that originated from inner membrane of nuclear envelope, were all noticed only in SlNPV-Btk combination sections. By 24h, only combination sections exhibited the highly necrotic lyses midgut appearance. Biocontrol agents and their combinations may provide an excellent alternative to conventional insecticides or at least minimize their use.
Key words: Culex pipiens, Nucleopolyhedrosis virus, Bacillus thuringiensis
2.Journal of Cell and Tissue Research 14(2): 4193- 4199 (2014)
Immunohistochemical expression of CD 44, CD24 and ESA in canine mammary tumours
Anjan Kumar, K.R., Sudhakar Rao, G.V., Balachandran, C., Murali Manohar, B., Dhanan Jaya Rao, C.G., Uzama, S. and Shammi, M.
Department of Veterinary Pathology, Veterinary College, Shimoga, KVAFSU; E. mail: drvet_anjan@hotmail.com
Abstract: Identification of Cancer stem cell lineage subpopulations of tumor cells within the tumor mass offer to identify targets for therapeutic interventions and a better understanding of carcinogenesis. The main objective this study was to evaluate the prognostic value of cancer stem cell lineage markers i.e., CD44, CD24 and Epithelial specific antigen (ESA) in canine mammary tumours using triple immunohistochemistry. A total of 40 surgically excised tissue samples of canine mammary tumours were obtained during the eight month study period. These tumors were classified according to WHO classification of mammary tumors of dog (2002), as Carcinoma – tubular, Carcinoma – Papillary cystic, Squamous cell carcinoma and Carcinosarcoma with tumor grades I & II respectively. Of 40 samples, 24 selected cases were subjected to triple immunohistochemistry using antibodies against CD 44, CD 24 and ESA. Two cases were positive for cancer stem cell phenotype i.e., CD44+/CD24-/low/ ESA+ showing colocalized expression of CD44, epithelial specific antigen (ESA) with membranous staining and absence or isolated staining for CD24, which correlated with both the cases showing distant metastases and reduced average survival period of 122 days, indicating poor survivability and distant metastasis. CD44+/CD24-/ESA+ expression was seen in carcinosarcoma (Grade I) indicating an epithelial mesenchymal transition and could favor metastasis. triple immunohistochemical expression of cancer stem cell phenotype i.e., CD44+/CD24-/ESA+ favored distant metastasis with poor prognosis and could be used as a prognostic indicator in canine mammary tumors as evident from this study.
Key words: Canine mammary tumor, CD 44, CD24, ESA
3.Journal of Cell and Tissue Research 14(2): 4201- 4209 (2014)
Histological study on the effect of formaldehyde on mice liver and kidney and possible protective role
of selenium
Treesh, S.A., El Jaafari, H.A., Darmun, E.I., Abu-Aisha, A.M., Alwaer, F.S., Eltubuly, R.A., Elghedamsi, M.and Aburawi, S.M.
Department of Histology and Medical Genetics, Faculty of Medicine; University of Tripoli, Tripoli, Libya.
E. mail: smaburawi@gmail.com
Abstract: This study was performed to identify the histological changes in the Liver and kidney resulting from formaldehyde injection and to investigate the possible protective role of selenium in repairing the hepatic and renal damage. Twenty four albino male mice were included in this study; intraperitoneal administration was adopted. Mice were divided into three groups (n=8). Group 1 mice received 5ml/kg. water for injection daily for five days (Control Group). Group 2 received 30mg/kg. of formaldehyde daily for five days. Group 3 (Cure Group) received formaldehyde daily for five days followed by daily administration of selenium (100μg/kg) for the next five days. Specimens of liver and kidney were evaluated histologically by Hematoxylin and Eosin and Mallory Trichrome stains. The result showed that formaldehyde exposure induced many histological changes in the liver and kidney. In formaldehyde treated group, the hepatocytes exhibited a vacuolated cytoplasm, dilated congested blood sinusoids, congested blood vessels and the von kupffer cells became more prominent. In the renal cortex, some glomeruli had dilated Bowman’s capsule space. The renal cortex exhibited a vacuolated cytoplasm of proximal convoluted tubules with loss of integrity of their brush border; also congested renal blood vessels and little damage of renal tubules were observed. Treatment with selenium led to a marked improvement of most of the previously mentioned changes as disappearance of cytoplasmic vacuolation of hepatocytes and preservation of renal tissue in a way that resembles the control. It can be concluded that, the short term exposure to formaldehyde can cause remarkable damage in the liver and kidney tissues; treatment with selenium as an antioxidant agent was effective in reversing the damage caused by formaldehyde in both liver and kidney.
Key words:, Liver, Kidney, Formaldehyde, Selenium
4.Journal of Cell and Tissue Research 14(2): 4211- 4218 (2014)
Biochemical and genotoxic effects of octylphenol in hepatomitochondrial fractions of freshwater fish,
Ooreochromis mossambicus
Sreedevi, N.V. and Chitra, K.C.
Department of Zoology, University of Calicut, Malappuram, District, Kerala, 673 635, India.E: mail: kcchitra@yahoo.com
Abstract: The environmental contaminants released into the aquatic ecosystem can disrupt the normal structure and functions of various organs in an organism thereby have an effect on its normal life. Octylphenol, one of the environmental contaminants, is known to perturb the normal functioning of aquatic life. In the present study octylphenol at 75µg/ L was exposed to freshwater fish, Oreochromis mossambicus for 7, 14 and 21 days and the biochemical changes and genotoxic potential was studied. During octylphenol treatment the body weights of the fish remained unchanged but the weight of liver and hepatosomatic index was decreased when compared to the control group. Octylphenol significantly increased mucous deposition all over the body in time-dependent manner. Biochemical estimation of the levels of lipid peroxidation, cytochrome oxidase and xanthine oxidase in hepatic mitochondrial fractions showed a significant increase at the end of treatment. This could be due to the generation of reactive oxygen species and energy demand in the mitochondrial fractions of hepatocytes. Exposure to octylphenol resulted in cytogenotoxicity as evidenced by fragmented apoptotic, bi-nucleated and sticky cells in micronuleus test and an increase in the number of colonies of the bacterium, Salmonella typhimurium, than that of control group. Therefore, octylphenol impairs antioxidant status and affects genotoxic potential when exposed chronically in freshwater fish O. mossambicus.
Key words: Octylphenol, Oreochromis mossambicus Hepatic mitochondria
5. Journal of Cell and Tissue Research 14(2): 4219- 4226 (2014)
Effect of bisphenol-A on the antioxidant defense system and its impact on the activity of succinate dehydrogenase in the gill of freshwater fish, Oreochromis mossambicus
Chitra, K. C. and Sajitha, R.
Department of Zoology, University of Calicut, Malappuram 673 635, Kerala. E. mail: kcchitra@yahoo.com,
Abstract: The study evaluated that bisphenol A induces oxidative stress to the exposed fishes by time-dependent reduction in the antioxidant enzymes and concomitant increase in the lipid peroxidation in the gill of Oreochromis. A significant reduction in the marker enzyme, succinate dehydrogenase (SDH) was also observed and this could be due to the impairment of aerobic metabolism in bisphenol- A treated fishes. Exposure to bisphenol- A for 20 days caused principal alterations in the gills of treated fishes as destruction of primary and secondary lamellae, upliftment of gill epithelium, hyperplasia and vasodialation in the lamellar epithelium. Thus the present study summarizes that the increased production of oxygen free radicals due to the exposure of bisphenol A inhibited the activities of antioxidant enzymes and succinate dehydrogenase thereby inducing oxidative stress and also destruct the normal architecture of gill in the fish.
Key words: Bisphenol A, Oxidative stress, Succinate dehydrogenase
6.Journal of Cell and Tissue Research 14(2): 4227- 4231 (2014)
Immunohistochemical and quantitative real time PCR expression of matrix metalloproteinases (MMPS) in canine mammary tumours
Shettar, M., Rao, S., Anjan Kumar, K.R., Purushottam, K.M., Rao, S., and Sathyanarayana, M.L.
Department of Pathology, Veterinary College, Hebbal, KVAFSU, Bangalore 560024; E. mail: shettardr@gmail.com
Abstract: Expression of matrix metalloproteinases (MMP2) was evaluated by immuno-histochemistry and quantitative Real time PCR. All the 19 malignant canine mammary tumours expressed MMP2 immunohistochemically as diffuse and granular reaction, which was restricted to cytoplasm of neoplastic epithelial cells, endothelial cells, fibroblasts, inflammatory cells especially macrophages and faintly in myoepithelial cells. The present study observed 16 malignant tumours that over expressed and three tumours that under expressed the MMP2. qRT PCR revealed a expression of MMP2 in all canine mammary tumours but expression was highly varied within tumour types and indicated that the tumours were in various stages in their progression towards attainment of malignancy with invasive and metastatic characteristics. Results of qRT PCR accorded well with the immunohistochemical results obtained with anti-human MMP2 monoclonal antibody (Clone K-20).
Key word: Metalloproteinases, Canine mammary tumors
7.Journal of Cell and Tissue Research 14(2): 4233- 4236 (2014)
Effect of bone morphogenetic protein-2(BMP-2) on sheep granulosa cell steriodogenic function
Vinod Kumar, D., Gulzar, R., Selvaraju, S., Nazar, S., Parthipans, Anand, Prasad, R.V., Jamuna, K.V.and Ravindra, J.P.
Department of Veterinary Anatomy and Histology, Veterinary College, Hebbal, Bangalore-560024.
Abstract: The bone morphogenetic protein-2 (BMP-2) has been implicated in follicle growth and steroidogenic functions of the ovary. The BMP-2 belongs to the BMPs subfamily of TGF-β super family. The present study was carried out in sheep granulosa cell culture to study the effect of different doses of BMP-2 0ng/ml (control group), 30ng/ml, 50ng/ml and 100ng/ml on granulosa cell steriodogenic function. For functional studies, granulosa cells were obtained from ovarian follicles 1–3 mm in diameter. The cells were cultured for 6 days in serum-free medium containing androstenedione and LR3-IGF-I. The medium was replaced for every two days and the spent medium was used to assess the concentration of estradiol 17-β in cultured granulosa cells by radioimmunoassay. The BMP-2 at all the three different doses increased the estradioal 17-β production when compared to control group of treatment (P<0.05), It was observed that even the lower doses tested (30 ng/ml) caused a significant increase in estradiol 17-β production by the granulosa cells as compared to the control (P<0.001), but no significant difference was observed among the different doses tested. It is concluded that all three different doses of BMP-2 were capable to increase the estradioal 17-β production without effecting the proliferation of granulosa cells in sheep.
Key words: BMP-2, Granulosa cell, Estradioal
8.Journal of Cell and Tissue Research 14(2): 4237- 4240 (2014)
Leutinising hormone receptor gene polymorphism and its association with post partum anestrus in
Murrah buffaloes
Kumar, R.,Gupta, M., Shafiq, S., Balhara, A.K. Sadeesh, E.M. and Singh. I.
Department of Veterinary Physiology & BiochemistryLala Lajpat Rai University of Veterinary and Animal Sciences, Hisar, Haryana-125004. E. mail: drrajesh7.vet@gmail.com
Abstract: To study polymorphism in the exon 11 of luteinizing hormone receptor (LHR) gene and to find its association with post partum anestrus (PPA) condition blood samples of 40 Murrah buffaloes in which 20 animals showing PPA (>120 days/gestation) and 20 animal showing normal cyclicity (<60 days/ gestation) for more than two consecutive gestation were taken in the present study. The genomic DNA was isolated and PCR was standardized to amplify partial exon 11 of luteinizing hormone receptor (LHR) gene 275bp by using primers. Amplified fragments of gene were sequenced and sequence variation was detected by doing multiple alignments. Among aligned population LHR gene showed one nucleotide deletion polymorphism at 243 bp in aligned sequence. Statistical analysis of this polymorphism between both the groups by chi square test revealed no significant association of this polymorphism with post partum anestrous condition in Murrah buffalo. Hence, role of polymorphism in exon 11 of luteinizing hormone receptor (LHR) gene with PPA cannot be established.
Key words: Murrah Buffalo, Luteinizing hormone receptor
9.Journal of Cell and Tissue Research 14(2): 4241- 4243 (2014)
Hematological profile in postpartum anestrous and normal cyclic Murrah buffaloes
Kumar, R., Gupta, M., Rose, M.K., Ghosh, M. and Singh, I.
Department of Veterinary Physiology and Biochemistry, Lala Lajpat Rai University of Veterinary
and Animal Sciences, Hisar, Haryana 125004. E-mail: drrajesh7.vet@gmail.com.
Abstract: Blood samples of twenty postpartum anestrous and twenty normal cyclic Murrah buffalo selected on the basis of their reproductive history and current status of reproductive organs were analyzed for haematological parameters. No significant differences between the postpartum anestrous (PPA) and normal cyclic animals were observed in red blood cell count (RBC), hemoglobin (Hb), packed cell volume (PCV) , platelet count, mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH), mean corpuscular hemoglobin concentration (MCHC), RBC distribution width (RdWc), MPV, platelet distribution width (PdWc), WBC, eosinophils, lymphocyte, monocyte, basophil, eosinophils (%), monocyte (%) and basophil (%). However, significantly (P<0.05) higher, neutrophil count as well as percentage of neutrophil and lymphocytes were recorded in the postpartum anestrous group buffaloes compared to that in the normal cyclic group indicating subclinical infection in the former groups. It can be concluded that subclinical bacterial infection might be one cause along with nutritional deficiency and heat stress for postpartum anestrous in buffaloes
Key word: Postpartum anestrous, Hematology, Buffalo
10. Journal of Cell and Tissue Research 14(2): 4245- 4250 (2014)
FecB genes in two different sheep breeds of Karnataka
Gadad, S.S., Jayashankar, M.R., Nagaraja, C.S., Satyanarayan, K. and Byregowda, S.M.
Department of Animal Genetics and Breeding, Veterinary College, Hebbal, Bangalore 560 024, Kvafsu,
Abstract: A study was conducted with the objective of analyzing the genetic variation of the FecB gene of Bannur and Tumkur sheep by using PCR-RFLP technique. Genomic DNA was isolated from blood samples of 50 each of Bannur and Tumkur sheep breeds. A 140 bp fragment of exon 6 of FecB gene were amplified. The sizes of the amplified products were uniform with no variation in size within the breeds of animals studied indicating conservation of DNA at this locus. Restriction endonucleases AvaII was used to detect the genetic variation in the FecB gene. AvaII restriction enzyme did not detect any polymorphism, revealing the absence of FecB mutation in Bannur and Tumkur sheep suggesting that all the animals were non-carriers for FecB mutation gene. The sequence analysis indicated high homology between present result and published FecB sequences. There was query coverage to the extent of 96% for Bannur and 98% for Tumkur sheep sequences, respectively and there was a 95% homology between the sequences of Bannur and Tumkur sheep. Two SNPs each at FecB locus were identified in Bannur and Tumkur sheep, respectively.
Key words: FecB Polymorphism, Sheeps
11. Journal of Cell and Tissue Research 14(2): 4251- 4254 (2014)
Toxic effect of azo dye (c.i. direct green 6) on blood parameters of freshwater fish Labeo rohita (ham.)
Barot, J, and Bahadur, A.
Department of Biosciences, V. N. South Gujarat University, Surat, Gujarat. E mail: anita26p@gmail.com,
Abstract: Haematotoxic effect of azo dye (Direct Green-6; DG 6; C.I. 30295) in widely consumed Indian major carp Labeo rohita (Ham.) was investigated. Fingerlings were exposed to three sublethal concentrations (2.5mg/l, 5.0mg/l and 10.0 mg/l) of DG-6 and their blood samples were collected and analysed at fixed intervals (5, 10, 15 days) followed by the exposure. A marked decrease in red blood corpuscles (RBC) count, haemoglobin (Hb) and packed cell volume (PCV) was noted. White blood corpuscles (WBC) count was also significantly decreased. Other related haematological indices such as mean corpuscular volume (MCV), mean corpuscular haemoglobin (MCH) and mean corpuscular haemoglobin concentration (MCHC) were calculated and these too showed a remarkable increase. These variations in the haematological parameters revealed erythrocytopenia, anaemic condition and leucocytopenia induced by intoxicated dye.
Key words: Azo dye, Haematotoxic effects, Labeo rohita
12. Journal of Cell and Tissue Research 14(2): 4255- 4258 (2014)
Identification of buffalo meat by polymerase chain reaction
Narendra Babu, R., Thulasi, G., Robinson, J.J., Abrahum, and Appa Rao, V.
Department of Meat Science and Technology, Madras Veterinary College, Chennai - 7, E. mail: nbabumst2@gmail.com
Abstract: Meat species identification involves a wide variety of techniques, including DNA based method. In present study, polymerase chain reaction (PCR) as means for authentication of origin of meat species have been used. Samples were taken as fresh, frozen, cooked and processed meat from indigenous non descript buffalo breeds for extraction of DNA, which was carried out once using simple lysis based extraction procedure and a commercial DNA extraction kit. Samples with OD ratio (260:280) between 1.7 to 1.9 were considered good in terms of concentration and purity and were used for PCR amplification using species specific oligonucleotide primers which were custom made based on conserved region of cytochrome b gene. Electrophoresis of amplified PCR products and gel documentation revealed 425 bp in 3% agarose gel. Added ingredients, additives and cooking temperature did not affect the efficacy of PCR amplification. Sequencing of purified PCR product was done by automated sequencing procedure using Big Dye terminator V 3.1 cycle and AB1 3700 DNA analyzer. The results of sequence analysis and comparison with the sequences for mitochondrial cytochrome b gene showed 100% identity with accession number D 88983.1 as published in National Centre for Biotechnology Information
Key words: Buffalo meat, Cyt b gene.
13. Journal of Cell and Tissue Research 14(2): 4259- 4265 (2014)
The evaluation of the antidiabetic effect of Coccinia indica leaves extract in streptozotocin induced diabetes in rats
Rao, S.,Rao, S., Manjunatha, K.P., Satyanarayan, M.L., Byregowda, S.M., Ramachandra, G., Sudharani, R., Shettar, M., Mundas, S. and Kalge, R.
Department of Veterinary Pathology, Veterinary College, Hebbal, KVAFSU, Bangalore. E. Mail : drraoiahvb@gmail.com,
Abstract: An alcoholic extract of coccinia indica leaves (200 mg/kg b w / day) was tested for its biochemical and pathomorphological effects in streptozotocin induced diabetes in rats. Experimental diabetes mellitus was induced in wister female rats with single buffer intraperitonial at 45 mg/kg. The rats were separated into five groups as follows: group 1 served as a normal control, grpoup-2 served as a diabetic control group-3 serves as a glibenclamide control, group-4 and group-5 serves as a diabetic rats administered with 200mg/kg Coccinia indica extract and Coccinia indica extract along with half dose of glibenclamide 300 ìg /kg body weight once daily for 90 days respectively. In diabetic animals there was a increase in the mean (± SE) serum levels of glucose, cholesterol, triglycerides, ALT and AST and a significant reduction in body weight and serum insulin. Coccinia indica leaves extract alone and in combination with glibenclamide half dose treated rats showed significant (Pd”0.001) reduction in the levels of serum glucose, cholesterol, triglycerides, ALT and AST from Day 3 post treatment till the end of the study with significant improvement in body weight and serum insulin levels (Pd”0.001) in diabetic animals. Coccinia indica leaves extract alone and with glibenclamide half dose also alleviated the damage caused by STZ morphologically in beta cells of islets of Langerhans and hepatocytes. However the combined treatment of Coccinia indica with glibenclamide half dose revealed a very good antidiabetic effect with reference to improvement in insulin level and beta cell number which indicated a synergistic effect between Coccinia indica and glibenclamide half dose.
Key words: Diabetes, Coccinia indica,
14. Journal of Cell and Tissue Research 14(2): 4267- 4272 (2014)
Forebrain embryonic zinc finger like gene poly-mormophism and its association with mastitis in Murrah buffalo
Dubey, T., Sangwan, M.L., Wani, S.A., Kumar, A., Gupta, A., Kumari, A. and Rana, V.
Department of Animal Biotechnology, Lala Lajpat Rai University of Veterinary and Animal Science,
Hisar 125 004, Hariyana.E. mail: tripurari.dube@gmail.com,
Abstract: Present investigation was carried out to study the Forebrain embryonic Zinc finger like gene polymorphism and its association with mastitis in Murrah buffaloes by using PCR-RFLP technique. Blood samples of 40 Murrah buffaloes were taken in which 20 animals were suffering from mastitis. The genomic DNA was isolated by phenol chloroform method. PCR was standardized to amplify partial region of (978bp) of gene using designed primers. PCR-RFLP analysis of the partial region of gene was studied using, Eae I and Msp I restriction endonucleases which gave restriction products of 978 bp, 655bp, 323 bp; 508bp, 320bp, 121bp & 29bp, respectively, in all the animals under study. Amplification of partial gene (978bp) using designed primers specific for cattle ortholog sequence signifies that the locus is conserved in cattle and buffaloes. The PCR-RFLP revealed same pattern viz. xy and yy of resolved digested fragments of FEZL gene with all the restriction enzymes used i.e. EaeI and MspI respectively in both group of animals i.e. healthy and mastitic. As monomorphic patterns were observed, hence FEZL gene association cannot be established with mastitis. So, further studies are needed with larger sample size, amplicon size and inclusion of more enzymes to get polymorphism in FEZL gene.Studies in this respect are in progress.
Key words: FEZL gene, Murrah buffalo, PCR-RFLP
15. Journal of Cell and Tissue Research 14(2): 4273- 4277 (2014)
Tissue distribution of different minerals and antioxidant status in various organs of Hyalomma anatolicum anatolicum (Acari: ixodidae) ticks
Ghosh, M., Sangwan, N., Sangwan, A. K., Kumar, R.and Gaur, R.S.
Department of Veterinary Physiology and Biochemistry, Lala Lajpat Rai University of Veterinary and Animal Sciences, Hisar 125004, Haryana. E. mail: ghosh.mayukh87@gmail.com,
Abstract: Ticks are obligate ectoparasites infesting varieties of vertebrate hosts. Hyalomma anatolicum anatolicum tick is one of the most important ectoparasite of cattle and buffaloes because it acts as the vector of tropical theileriosis causing huge economic loss in terms of production in India. It also takes large amount of blood from their hosts causing production loss. During their prolonged feeding on their vertebrate hosts, the release of different reactive oxygen and nitrogen species by exogenous and endogenous means can potentially induce oxidative damage to ticks. So ticks evolved an effective free radical scavenging and antioxidant defense system for their successful feeding of a blood meal. Therefore, the present study was aimed to explore the roles of free radical scavenging activity and the level of different antioxidants and some micro & micro elements by their quantitative estimation in different organs of engorging female Hyalomma a. anatolicum ticks. In current work, nitric oxide scavenging and superoxide dismutase (SOD) activities, reduced glutathione (GSH) concentrations and level of some micro & macro elements like- copper (Cu), zinc (Zn), manganese (Mn), iron (Fe), calcium (Ca) and magnesium (Mg) were estimated in different organs (salivary gland, gut and ovary) of engorging female H. anatolicum ticks. Variations in antioxidant activities and metal ion concentrations were observed in different organs of H. anatolicumticks that will help in better understanding about tick biology and to develop improved tick control strategies.
Key words: Hyalomma anatolicum anatolicum, Antioxidants; Free radicals
16. Journal of Cell and Tissue Research 14(2): 4279- 4282 (2014)
Molecular finger printing of mitochondrial 12s rRNA and 16s rRNA gene of the indigenous pig of Assam
Saikia, D.P. Kalita, D.J. Borah, P., Zaman, G.U., Dutta, R. and Saikia, B.
Department of Animal Biotechnology, College of Veterinary Science, Assam Agricultural University, Khanapara, Guwahati 781022 Assam. E. mail: saikiadeep17@gmail.com, Cell: 08859875506
Abstract: Polymerase chain reaction (PCR) in combination with restriction fragment length polymorphism (RFLP) is the most efficient and preferred method for characterization of different species in terms of the applicability and detection limit. The present investigation was carried out with an aim to develop a molecular fingerprint of mitochondrial 12S rRNA and 16S rRNA gene of the indigenous pigs of Assam. Ten random blood samples were collected from different places of Assam for mitochondrial DNA extraction. The extracted DNA was used for the amplification of 12S rRNA and 16S rRNA gene by universal primers. The size of the amplicon was 456bp and 600bp respectively for both the gene. RFLP studies were carried out with restriction enzymes viz. AluI and ApoI respectively for both the gene. To confirm the variations at nucleotide level sequencing of the PCR product and analysis of the sequence result was performed.
Key words: Mitochondrial DNA, 12S rRNA gene, 16S rRNA gene, Indigenous pigs
17. Journal of Cell and Tissue Research 14(2): 4283- 4285 (2014)
Impact of vitamin e and c on nitrogen dioxide gas induced alteration in blood coagulation factors of male Albino rat
Agarwal, A. and Khan, R.
Department of Zoology, School of Life Sciences, Khandari Campus, Dr. B.R.Ambedkar University,
Agra 282 003, U.P. E. mail: bajahidtrue@gmail.com
Abstract: In the present investigation, the effect of nitrogen dioxide exposure (50ppm) and role of vitamins E (2.5mg/rat) and vitamin C (5mg/rat) supplementation in combination on coagulation factors have been reported inmale albino rats. An increase in coagulation factors viz. Prothombintime (PT), Partial thromboplastin time (PTT), Activated partial thromboplastin time (APTT), Thrombin time (TT) and platelet count (PLT), while decrease in fibrinogen deficiency plasma (FDP) level was reported after supplementation of vitamin E and C in comparison to nitrogen dioxide gas exposed rats. Present study suggests that administration of vitamin E and vitamin C in combination shows protective and beneficial effect to blood coagulation factors of albino rats against nitrogen dioxide gas toxicity.
Key words: NO2 gas, Vitamin (E+C), Coagulation factors
18. Journal of Cell and Tissue Research 14(2): 4287- 4291 (2014)
Adventitious shoot regeneration in Safflower (Carthamus tin ctorius l.) using TDZ×NAA
Mohite, N.R., Dudhare, M.S., Jadhav, P.V., Moharil, M.P. and Deshmukh, A.G.
Biotechnology centre, Dr. Punjabrao Deshmukh Krishi Vidyapeeth , Akola- 444104, Maharashtra.
E. mail: mahendra_s_d@yahoo.com,
Abstract: Carthamus tinctorius L. commonly known as safflower is an important oilseed crop cultivated in several parts of India and it is widely exploited for its medicinal properties. An efficient protocol for in vitro shoot regeneration in two varieties of safflower, namely AKS-207 and PKV-pink was developed. Cotyledonary leaf segments and cotyledonary nodes were used in the present investigation and those were obtained by in-vitro germinating seedlings of 10-days-old. Both explant cultures were raised on Murashige and Skoog’s (MS) medium supplemented with different concentrations of either TDZ or in combination with NAA. 100% percentage of shoot induction within 8-9 days, followed by high frequency of shooting was observed in AKS-207 with cotyledonary node used as explant on MS medium fortified with TDZ + NAA (2.0 + 0.1mg/l).
Key words: Carthamus tinctorius L., In vitro shoot regeneration
19. Journal of Cell and Tissue Research 14(2): 4293-4296 (2014)
Rapid one step protocol for in vitro regeneration of Bacopa monnieri (L.)
Naik, P.M., Patil, B.R., Kotagi, K.S., Kazi, A.M., Lokesh, H. and Kamplikoppa, S.G.
Department of Genetics and Plant Breeding, University of Agricultural Sciences, Dharwad-580005;
Karnataka. E. mail: patilbhuvaneshwara@gmail.com
Abstract: A cost-effective and rapid in vitro regeneration protocol was described in the present work for large scale in vitro propagation of a valuable medicinal herb Bacopa monnieri (L.) by leaf, node and internode segments isolated from field grown mature plants. Leaf, node and internode explants were grown on Murashige and Skoog medium supplemented 2 mg/l kinetin (KN) for 8 weeks. Highest rate of shoot regeneration was observed in leaf explants (72.25 shoots/explants). The fresh and dry weight of shoots was also highest with this treatment (4.050 g fresh and 0.208 g dry wt). Maximum number of rooting was seen in leaf explant (44.25 roots/explants) supplemented 2 mg/l KN. The resulted plantlets were successfully transferred to soil.
Key words: Bacopamonnieri, In vitro regeneration
20. Journal of Cell and Tissue Research 14(2): 4297-4302 (2014)
In vitro multiple shoot induction and plantlet formation in tomato
Archana, K., Archana, P., Mahitha, B. and Rama Swamy, N.
Plant Biotechnology Research Laboratory, Department of Biotechnology, Kakatiya University,
Warangal 506009. E. mail: swamynr.dr@gmail.com,
Abstract: In present contribution the effect of different plant growth regulators (PGRs) on in vitro direct plant regeneration of tomato cv Arka Vikas from primary leaf explants has been reported. Primary leaf explants excised from in vitro grown seedlings (15 days old) were cultured on Murashige and Skoog’s (MS) medium supplemented with different concentrations (0.5-4.0 mg/L) of 6- Benzyl amino purine (BAP) and Kinetin (KIN ) individually and also in combination with 0.5mg/L Indole-3-acetic acid (IAA). Primary leaf explants cultured on medium containing BAP (2.0 mg/L) in combination with IAA (0.5 mg/L ) induced maximum number of multiple shoots (15.8±0.04) with increased mean shoot length (4.3 ± 0.09). Addition of IAA to MS medium supplemented with BAP triggered for induction of more number of multiple shoots from primary leaf explants compared to single cytokinin. The elongated shoots were excised and rooted on MS medium supplemented with IAA (0.5- 2.0 mg/L). The in vitro rooted plantlets were acclimatized and the survival percentage was found to be 80%. The protocol developed in the present investigation can be used for genetic transformation studies in tomato cv Arka Vikas for introducing a novel gene.
Key words: Multiple shoot induction, Tomato
21. Journal of Cell and Tissue Research 14(2): 4303-4308 (2014)
Antioxidant potential in seeds of Coriandrum sativum: An in vitro study
Kaur, D., Chugh, V. and Soni, G.
Department of Biochemistry, Punjab Agricultural University, Ludhiana. E. mail: dasmeet.kaur@gmail.com
Abstract: In recent years, natural antioxidants in food and other biological materials have drawn considerable concern due to their presumed safety, nutritional and therapeutic value that offers protection against a range of non-communicable diseases and ageing. The present study focuses on evaluation of Coriandrum sativum (Coriander) varieties for their antioxygenic potential using different in vitro free radical scavenging models. The methanolic extracts of local and simco varieties of coriander were found to be appreciably effective in scavenging hydroxyl radical generated by Fenton reaction (EC50=38 and 18μg/ml), super oxide radical generated by photoreduction of riboflavin (EC50=553.50 and 668.67μg/ml), and nitric oxide radical generated in vitro from sodium nitroprusside (EC50=180 and 227μg/ml) in a concentration dependent manner. In contrast, methanol: hexane extracts of both the varieties of coriander were found to be ineffective in quenching hydroxyl radical and revealed only moderate activity in quenching super oxide radical (EC50=712 and 832μg/ml) and nitric oxide radical (EC50=569 and 703μg/ml). However, hexane extracts exhibited no appreciable affects at either of the concentrations in all the three models. Instead, it showed pro-oxidant activity at higher concentrations. Above mentioned in vitro models proved Local variety to possess better antioxygenic potential. Likewise, the inhibition of in vitro linoleic acid peroxidation for longer period of incubation by methanolic extract (superior to ascorbic acid, a standard antioxidant) and lipid peroxidation in rat liver homogenate (EC50=383µg/ml) also indicated good antioxygenic potential of Local variety of coriander.
Key words: Coriander, Antioxidant activity, oxidative stress
22. Journal of Cell and Tissue Research 14(2): 4309-4314 (2014)
Establishment of an in vitro micropropagation protocol for a medicinal herb Tylophora indica
Anjum, A., Narula, A., Khan, M, A.,and Kamaluddin
Department of Biotechnology, Jamia Hamdard, New Delhi-110062, E. mail: anwarkhan@skuastkashmir.ac.in,
Abstract: An efficient micropropagation protocol has been developed for in vitro propagation of endangered medicinal herb (Tylophora indica) using leaf and nodal segment as an explant collected from one year old mature plant. The medium used for the present study was 3% MS (Murashige and Skoog) medium supplemented with different concentrations and combinations of growth hormones (IAA, BA, NAA, Kn).The response was periodically recorded after 4, 8 and 12 weeks. The concentration of plant growth regulators and explant types exhibited discrete roles in the efficiency of plant regeneration. The regeneration potentiality of leaf segment was better on medium MS + NAA (0.5mg/l) + BA (2.0 mg/l). Nodal segment showed better regeneration than leaf and started to respond after 4 weeks. Maximum regeneration was obtained on MS + IAA (0.5mg/l) + Kn (2.0mg/l). The study demonstrated a propagation route via leaf and nodal segment in T. indica, which could be useful for large scale multiplication of this medicinal plant.
Key words: Tylophora indica, Micropropagation
23. Journal of Cell and Tissue Research 14(2): 4315-4321 (2014)
In vitro callus approach in selection for drought tolerance in bread wheat and its relation to yield performance under field drought conditions
Gupta, N., Bains, N.S. and Thind, S.K.
Department of Botany, Punjab Agricultural University, Ludhiana 141004. E. mail: nehagupta.pau@gmail.com
Abstract: In present study, bread wheat (Triticum aestivum L.) genotypes were evaluated for drought tolerance under in vitro and field drought conditions. At in vitro level, genotypic differences were significant for callus fresh weight, growth index and relative water content on 0%, 5% and 10% PEG selection media assessing that in vitro callus growth was affected with higher levels of PEG. DBW 17 followed by PBW 175 and PBW 550 were superior in sequence for in vitro PEG tolerance. In contrast, genotypes BW 9183, BW 9007 and BW L0089 exhibited severe reductions in calli fresh weight and growth index under PEG induced drought stress. At field level, significant reductions in ear head/m2, number of grains/spike, thousand grain weight resulted in grain yield loss under drought stress. For drought tolerance, the genotypic response observed under field conditions was reverse of the demarcations observed under in vitro stress conditions. Genotype DBW 17 and PBW 550 which exhibited tolerance to drought stress under in vitro conditions, were sensitive at field level. In contrast, genotypes BWL0089 and BW 9007 which performed poorly under in vitro conditions, were tolerant to field based stress. Thus, in vitro screening can not be used as a direct strategy in selection of drought tolerant genotypes for field conditions.
Key words: Drought tolerance, Triticum aestivum, Callus growth
24. Journal of Cell and Tissue Research 14(2): 4323-4331 (2014)
Effect of different plant hormones on node culture in pointed gourd
Ahmad, T., Kumar, V., Patel, N. L., Kapadia C.V., Suthar, K. P. and Singh, D.
Department of Plant Molecular Biology and Biotechnology, ASPEE College of Horticulture and
Forestry, Navsari Agricultural University, Navsari 396450, Gujarat. E mail: chintan_bt@yahoo.co.in
Abstract: In India pointed gourd (Trichosanthes dioca Roxb.) is extensively cultivated in eastern Uttar Pradesh, Bihar and West Bengal and to some extent in Assam, Orissa, Madhya Pradesh, Maharashtra and Gujarat. Propagation of pointed gourd from seed is not desirable due to poor germination as well as dioecism which results in 50% unproductive male plants. Traditionally pointed gourd is multiplied through stem cuttings and root suckers. For planting one hectare land, 6,000-7,500 cuttings are required, however cuttings are not easily available and therefore tissue culture offers a promise for the bulk supply of planting material which can be transplanted in a proper sex ratio in the field. Effect of various treatments of plant hormones supplemented in MS medium was examined for node culture in four genotypes of pointed gourd viz. Calcutti Female, Calcutti Male, Hybrid Female, and Hybrid Male. Four different parameters viz. (i) per cent establishment, (ii) number of shoots per culture, (iii) shoot length and (iv) number of nodes per culture was studied. The medium supplemented with 1.0 mg/l IAA + 0.2 mg/l BA showed longest shoot with better and quicker establishment and little or no root formation in all the genotypes of pointed gourd. This was found to be best culture establishment medium. Among different genotypes, in vitro response of Calcutti Female was best followed by Hybrid Female, Calcutti Male and Hybrid Male. Of the various treatment of plant hormones examined in MS medium, the same treatment 1.0 mg/l IAA + 0.2 mg/l BA was found to be best shoot proliferation medium for continuous sub culturing. Rooting of in vitro shoots in pointed gourd was not a problem as rooting occurred in many treatments during shoot establishment itself. Half strength MS medium supplemented with 0.1mg/l IAA + 0.2 mg/l IBA was found most effective for rooting of in vitro raised shoots of pointed gourd as it gave highest number of longest root with maximum frequency.
Key words: Node culture, pointed gourd, Trichosanthes dioca Roxb
25. Journal of Cell and Tissue Research 14(2): 4333-4338 (2014)
Micropropagation strategies for conservation of endangered medicinal plant Withania somnifera
(l.) dunal
Sharma, N., Sharma, M.D., Dhiman, M., and Koshy, E.P.
Department of Tissue Engineering, Jacob School of Biotechnology, SHIATS Allahabad, Uttar Pradesh,
E.. mail: nishesh21@gmail.com
Abstract: Withania somnifera commonly called as Ashwagandha is an important medicinal plant which has become endangered due to over utilization. In respect of endangered status of W. sominfera in vitro culture studies have gained importance for conservation of the plant. Hence, present work avails a rapid and effective protocol for conservation and mass propagation of the species through tissue culture technique by utilizing nodal segments as explants onto MS medium containing NAA and Kn. 100% cultures showed multiple shoot regeneration, with a maximum of 7 shoots and average shoot number of 3.4 ± 0.33 per culture onto 10 µm Kn. 100% cultures exhibited callus induction with 92.1% cultures showing shoot bud initiation in MS media fortified with 4 µm NAA and 10 µm Kn. A maximum of 90.6 ± 0.20 % cultures were successfully rooted onto ½ MS medium fortified with 20 µm IBA. Regenerated plants were acclimatized and about 53.4 % plants survived during acclimatization and were transferred to field.
Key words: Withania somnifera, Micropropagation, Napthalic acetic acid
26. Journal of Cell and Tissue Research 14(2): 4339-4344 (2014)
Adventitious shoot regeneration from hypocotyls of Wrightia arborea (Dennst.) Mabb.: An endangered toy wood species
Nagalakshmi, M., Vishwanath, S. and Viswanath, S.
Tree improvement and Genetics division, Institute of Wood science and Technology, Malleshwaram
Abstract: The species Wrightia arborea (Dennst.) Mabb is a multipurpose tree with economically important timber and therapeutic significance. It is highly exploited in toy making industry for its ivory like wood. Due to its over exploitation, the species has become endangered. The present investigation reports on a protocol for conservation of W. arborea through direct organogenesis from hypocotyl. The ability of hypocotyl segments to produce shoot buds varied depending upon their position on the seedling. Hypocotyl segments obtained from closer to the root were highly responsive and produced a maximum of 12 shoots per explant in MS medium supplemented with BAP (0.5 mg/l), AdSO4 (1mg/l). MS medium with BAP (0.5mg/l), IBA (1mg/l) and AdSO4 (1mg/l) was proved to be the best for proliferation and elongation. Before rooting, thin elongated shoots were maintained for one passage in MS medium with BAP (0.5mg/l), IBA (1mg/l) and AdSO4 (1mg/l) and charcoal (50mg/l) to obtain sturdy shoots. Sturdy individual shoots were transferred to quarter strength MS medium with charcoal (50mg/l) devoid of growth regulators for rooting. Thus regenerated plantlets were primary hardened successfully in sterilized vermiculite for a period of 45 days and then transferred to poly bags.
Key words: Wrightia arborea, adventitious regeneration, Hypocotyl.
27. Journal of Cell and Tissue Research 14(2): 4345-4350 (2014)
Efficient in vitro micro propagation of sugar cane (Saccharum spp. complex CV. CO 99004) through callus culture
Gadakh, S.S., Patel, D.U., Narwade, A.V., Mali, S.C., Mehta, R. and Singh, D.
Department of Genetics and Plant Breeding, N.M. College of Agriculture, Navsari Agricultural
University, Navsari – 396 450, Gujarat. E. mail: gadakhsuraj@gmail.com, Cell: 07588079791
Abstract: Standardization of protocol for induction of callus and regeneration of plantlets was established through in vitro culture using meristem of Sugarcane (Saccharum spp. Complex, cv. Co 99004) as an explant. The multiple shoot regeneration at various frequencies was observed by using different concentrations and combinations of growth regulators. The highest percentage of callus induction was observed in MS medium supplemented with 4 mg/l, 2,4-D. The best response in terms of multiple shoot induction was observed on MS medium supplemented with NAA 0.5 mg/l + BAP 1 mg/l. More profuse rooting observed when in vitro shoot lets were inoculated on to the half-strength MS basal + 2 % sucrose media supplemented with 0.5mg/l NAA + 1mg/l IBA. Rooted shoots were transplanted in the green house for hardening and best hardening response was observed in potting mixture containing Clay + Sand + Bio-compost (1:1:1 ratio) in primary and secondary hardening.
Key words: Micro propagation, Sugar cane
28. Journal of Cell and Tissue Research 14(2): 4351-4356 (2014)
Differential cultural responses of wheat (Triticum aestivum l.) with different explants
Gill, A. K., Gosal, S. S. and Sah, S. K.
School of Agricultural Biotechnology, Directorate of Research, Punjab Agricultural
University, Ludhiana 141 004, Punjab. E. mail: amangill1189@gmail.com,
Abstract: Different explants viz. immature embryos, immature inflorescences and mature grains of two bread wheat varieties viz. HD2967 and Bobwhite were used as explant source for establishing tissue cultures. Explants cultured on MS medium supplemented with 2,4-D (2.0 mgL-1) showed best callus induction except for mature grains which showed best callusing on MS medium supplemented with 2,4-D (3.0 mgL-1). Highest shoot regeneration from mature grain-derived calli was observed on MS medium supplemented with TDZ (0.3 mgL-1) in HD 2967 and in Bobwhite optimum regeneration was observed on MS medium supplemented with TDZ (0.2 mgL-1). Highest shoot regeneration from immature inflorescence-derived calli was observed on MS medium supplemented with TDZ (0.1 mgL-1). Optimum shoot regeneration from immature embryo-derived calli was observed on TDZ (0.2 mgL-1) containing medium. From the results of this study immature embryos were selected to be best explants with best callus induction and subsequent plant regeneration, although best embryogenesis was observed in calli derived from immature inflorescences. TDZ was found to be superior in regeneration over BAP and IAA combinations.
Key words: Triticum aestivum, Embryogenesis, Shoot regeneration
29. Journal of Cell and Tissue Research 14(2): 4357-4362 (2014)
Optimization for the most organogenic responsive combinations of plant growth regulators through in vitro regeneration of tomato cv Pusa dwarf
Patel, N., Kansara, R.,Mahatama, M. and Mahatma, L.
Department of Plant Molecular Biology & Biotechnology, Navsari Agricultural University, Navsari-396450, Gujarat. E. mail: nafisa.z.patel@gmail.com
Abstract: Successful in vitro plant regeneration protocol has been optimized for the parental variety of tomato (cv Pusa dwarf) through leaves. Shoot Regeneration was obtained directly from leaves as well as indirectly through callusing. Leaves were surface sterilized and cultured on Murashige and Skoog (MS) medium supplemented with various concentrations of cytokinins and auxins either alone or in combination. The combination of BAP and Kinetin with IAA gave maximum callus formation and shoot regenearation. The highest number of shoots as well as the maximum shoot length was seen with 2 mg/l BAP + 1 mg/l IAA. The other combinations of growth regulators with 2 mg/l kinetin + BAP 0.20 mg/l + 0.30 mg/l IAA seems to be an effective combination giving 57.78% multiplication. The minimum number of days and the maximum root formation was found in the Half MS medium fortified with 0.50 mg/l IBA. The maximum survival (61 %) of plantlets with minimum days for new sprouting (11.25 days) was reported in Vermicompost: Soil mixture in the hardening process.
Key words:Lycopersicon esculantum, Growth regulators, Pusa Dwarf
30. Journal of Cell and Tissue Research 14(2): 4363-4366 (2014)
Effect of growth regulators on micro propagation of potato cultivars
Kaur, M., Kaur, R., Sharma, C., Kaur, N. and Kaur, A.
School of Agricultural Biotechnology, Punjab Agricultural University, Ludhiana, Punjab.
E. mail: chhaya_uhf@yahoo.co.in
Abstract: The effect of growth regulators on micropropagation (in vitro) of three potato cultivars (Solanum tuberosum L.) were evaluated. In the present study nodal explants of potato cultivars (Diamont, 1533 and Kufri Badshah) were cultured on MS basal medium supplemented with different hormonal combinations (BAP and GA3). Out of four different media combinations, MS + BAP (1.0 mg/l) gave maximum shoot regeneration on all the three cultivars of potato. Successful rooting was achieved by placing the shoots on half strength MS basal medium. The combination of sand: soil (1:1) was the best for plant acclimatization as 90% of the plants survived and became established.
Key words: Potato, Nodal explants, Micropropagation
31. Journal of Cell and Tissue Research 14(2): 4367-4372 (2014)
Evaluation of induced mastitis in rabbits based on clinical parameters, direct microscopic somatic cell count and viable counts of E. coli in milk
Ravikumar, P., Rao, S., Byre Gowda , S.M., Satyanarayana , M.L.,Isloor, S., Rajeshwari, Y.B. and Jayaramu, G.M.
Dept of Veterinary Pathology, Veterinary College, Hassan, Karnataka 573202;
E. mail: ravikumarputtamallappa@gmail.com.
Abstract: Mastitis was induced in lactating rabbits by inoculating E.coli strains isolated form bovine mastitis cases. Six rabbits each were infected with E.coli strains O9(36) and O147 isolated from bovine mastitis cases. The milk samples were collected from the rabbits at the interval of 24 hours after carefully recording the clinical signs of mastitis. Both O9(36) and O147 strains of E.coli successfully induced clinical mastitis in rabbits by 24 hrs as shown by rise in body temperature, dullness, depression and inflammation of the mammary glands characterized by hyperemia, enlargement, heat and pain on palpation. The severity of clinical signs reduced gradually from 3rd day post inoculation. The SCC of normal rabbits ranged from 0.15- 0.18X106 /ml of milk. There was a drastic rise in the SCC of rabbits infected with either O9(36) or O147 strain of E.coli by 24 hours PI which reached a peak at 48 hours PI. The peak SCC values observed in the infected rabbits at 48 hours PI at the infective dose of 106 CFU/ml were 7.60 X106 and 9.86 X106 /ml respectively in the rabbits infected with O9(36) and O147 strains. This increase in SCC was characterized mainly by a phenomenal increase in the number of desquamated epithelial cells and PMNs at 24 & 48 hours PI. The SCC counts of infected rabbits showed a decreasing trend from 72 hours PI and observed to be replaced by mononuclear cells. The mean viable counts of E.coli in the milk of rabbits infected with either O9(36) or O147 E. coli strain showed a rapid rise at 24 hours PI which reached a peak at 48 hours PI at all the infective dose levels (104 cfu to 107 cfu) and tended to decrease from 72 hours till 144 hours PI. The mean viable counts for 104 cfu and 105cfu dose levels were 3.35 × 106 cells/ml & 5.48 × 106 cells/ml respectively at 24 hours PI for the O9(36) infected rabbits and 2.47 × 106 cells/ml & 5.26 × 106 cells/ml respectively for the O147 infected rabbits. The mean viable counts and SCC were significantly higher (P < 0.01) at all the infective dose levels compared to that of PBS infused glands. Based on the clinical involvement, SCC and viable counts of E.coli in milk, it was concluded that the bovine mastitis strains of E.coli successfully induced clinical mastitis in rabbits.
Key words: E. coli, Mastitis, Rabbits
32. Journal of Cell and Tissue Research 14(2): 4373-4376 (2014)
Evaluation of DNA extraction protocol for detection of Salmonella from animal foods
Porteen, K., Gunaseelan, L., Bhanu Rekha, V., Pawar, G.R. and Balakrishnan, S.
Department of Veterinary Public Health and Epidemiology, Madras Veterinary College, Chennai- 600007. E mail: rajavet2002@gmail.com
Abstract: Detection of Salmonella requires rapid and specific methods for isolation, identification and enumeration but classical microbial procedures for the detection of Salmonella from foods are laborious and time consuming. In recent years PCR is a rapid and powerful molecular biology tool that is able to detect positive samples with extremely high sensitivity, which makes it a promising technique applicable to monitoring programs. However, the use of PCR directly with variety of animal food samples has been very limited because of the large variety of substances that might inhibit the reaction. An appropriate DNA-extraction step should be able to remove or, at least, dilute such inhibitory compounds to levels that do not compromise PCR performance. DNA-extraction methods using lysozyme, proteinase K and phenol-chloroform are time-consuming, expensive and laborious. The use of commercial kits based methods is effective but expensive. Considering the above points the present study was done to evaluate a cost effective DNA isolation method (alkaline lysis PEG method (AL-PEG)1 from foods of animal origin and its suitability in PCR is presented.
Key words: Salmonella, Polymerase Chain Reaction, DNA Extraction, Alkaline lysis
33. Journal of Cell and Tissue Research 14(2): 4377-4384 (2014)
Multimycotoxin analysis using LC-MS/MS and sub chronic toxicity study of Fusarium semitectum fungus isolated from groundnut hay
Vinay, P.T.,Shridhar, N.B., Sanganal, J.S., Narayanaswamy, H.D., Isloor, S.and Chitra, B.M.
Department of Pharmacology and Toxicology, Veterinary College, KVAFSU, Bidar-585226;
Abstract: The present study was conducted to know the cause of the obscure disease found in the cattle of south Karnataka region since 2005. This study confirmed the disease in cattle, which occurred after feeding of the contaminated groundnut hay. Ailing cattle were showing the clinical signs of colic, tenesmus, bleeding from natural orifices, central nervous system abnormality symptoms, anorexia, chronic recumbancy and death after few days. Fungus was isolated from the groundnut hay, identified, characterized by molecular methods. The fungus contaminated wheat materials were subjected to multimycotoxin analysis using LC-MS/MS method. Apicidin and beauvericin mycotoxins found in high concentrations. To confirm further, toxicity studies were conducted in rats. Clinical signs of toxicity were observed. Body weight, blood clotting time, postmortem findings, histopathological changes, hematological parameters, and biochemical parameters abnormality were recorded and correlated. The present study indicated the toxic feature of the culture filtrates/ fungi infected wheat material isolated from ground nut hay in rats. This study helped the farming community by identifying cause of the death of the cattle and reduced further mortality.
Key words: Fusarium semitectum, Fungal isolate, Groundnut hay
34. Journal of Cell and Tissue Research 14(2): 4385-4390 (2014)
Assessment of biochemical diversity in diverse maize genotypes using isozyme markers
Sharma, R., Maloo, S. R. and Joshi, A.
Department of Molecular Biology and Biotechnology, Rajasthan College of Agriculture, Maharana Pratap University of Agriculture and Technology, Udaipur 313001, Rajasthan. E. mail: radhebiotech88@gmail.com. Cell: 09660235287
Abstract: Maize is considered to be one of the important coarse cereal crop in India. It is also known as “queen of cereals” due to their wider adaptability and higher production potentials. In present study twenty maize genotypes were used for biochemical diversity analysis using three enzymes viz., esterase, peroxidase and superoxide dismutase. These enzymes were screened by native polyacrylamide gel electrophoresis (PAGE) technique to study the variation in isozymatic patterns. Only nine bands were resolved for young (28 days after sowing) leaf samples. The isozyme profile for esterase showed two bands having Rm value of 0.1 and 2.5. Whereas, peroxidase showed two activity zones having Rm value of 0.25 and 0.44 and superoxide dismutase showed four bands having Rm value 0.1, 0.29, 0.41 and 0.55, respectively. A total 8 alleles were generated by three isozyme. Easily resolved and bright isozyme bands were only counted. Superoxide dismutase isozyme showed 63% polymorphism could be attributed to environmental condition. The pooled data for Jaccard’s similarity coefficient values revealed that the genetic distance derived from isozyme analysis was very low (0-11%) due to the small number of polymorphic bands. The above study inferred that all genotypes might have some ancestral/phylogenic relationship due to migration/inflow of genes.
Key words: Maize, Isozyme, Genetic diversity
35. Journal of Cell and Tissue Research 14(2): 4391-4394 (2014)
Canine persistent wound pathogens and their association with biofilm formation
Padhy, A. and Sahu, A.R.
Department of Veterinary Microbiology, College of Veterinary Science and Animal Husbandry,
Bhubaneswar 751003, Odisha. E. mail: dearlita123@gmail.com,
Abstract: The persistency of wound infections and treatment failure for prolonged period of time has shown a major association with biofilm formation in many animals. Most of the canine persistent wounds also show biofilm formation. 172 different types of persistent wound samples were taken for isolation and identification of bacterial isolates and then processed for biofilm production by Tube and TCP method. The wound samples showed that they were polymicrobial in nature and nearly 50% of isolates showed the biofilm formation suggesting that biofilm formation by bacterial isolates in dog wounds was the cause of their persistency and treatment failure.
Key words: Persistent wound, Biofilm
36. Journal of Cell and Tissue Research 14(2): 4495-4403 (2014)
Induction of systemic acquired resistance in Arachis hypogaeal. by Nspergillus flavus derived elicitors
Nandini, D.and Bariya, H.
Department of Biotechnology, Shri A. N. Patel Post Graduate Institute, Sardar Patel University,
Anand 388110, Gujarat. E. mail: durgeshnandinibt@gmail.com,
Abstract: Aflatoxin production caused by Aspergillus flavus in peanut (Arachis hypogaea L.) seeds is a serious problem. Fungal components of Aspergillus flavus in form of fungal culture filtrate and mycelial cell wall fractions were used as elicitors to provoke the defense response in four different varieties (J-11, GG-20, TG-26 and TPG-41) of Arachis hypogaea L. Biochemical changes i.e., salicylic acid as signaling molecule and marker enzymes such as peroxidase, phenyl alanine ammonia lyase, β-1, 3 glucanase and lignin content. There was a substantial fold count increase in peroxidase, phenylalanine ammonia lyase activity, salicylic acid and β-1, 3 glucanase and lignin content in fungal culture filtrate and mycelial cell wall fractions treated plants of all varieties of groundnut than in control plants. Higher enzyme contant was observed in fungal culture filtrate treated plants than mycelial cell wall fractions treated plants. Levels of aflatoxins were negligible in all FCF treated four varieties of groundnut plants stored seeds as compared to mycelial cell wall fractions and control plants seeds. The present results indicate that the use of fungal components (fungal culture filtrate and mycelial cell wall fractions) had successfully induced systemic resistance in four different varieties of groundnut plants against Aspergillus flavus.
Key words: Aflatoxin, Aspergillus flavus. Arachis hypogaea L
.
37. Journal of Cell and Tissue Research 14(2): 4405-4408 (2014)
Identification of biofilm and extended spectrum β-lactamases (ESBL) producing isolate of Klebsiella pneumonia from Tamilnadu
Prakash, B., Mathew, L.P., Narendrababu. R., Perumal, P., Geandeal, M.A.and Jagathesh, P.
Department of Biotechnology, PGP College of Arts & Science, Namakkal 637207, India; E. mail: nbabumst2@gmail.com
Abstract: Prevention of food-borne illness is an important public health concern worldwide. Consumption of raw or undercooked unhigeneic meat and eggs linked to health risk to individuals with compromised immune system. In the recent years, multi drug resistant Escherichia coli and Klebsiella pneumoniae strains pose a great threat to many countries. The aim of the present study was to evaluate the presence of drug resistant K. pneumonia from local meat chains. Chicken, beef, goat tissue samples from small scale abattoir and commercially available processed meat sausages were tested for the presence of multi drug resistant K. pneumonia. Among the tested meat samples, highest occurrence of β-lactamase producing K. pneumonia was observed in goat meat sample (70%), bovine (60%) and chicken (10%). The prevalence of extended-spectrum β-lactamase genes identified by the amplification of SHV, TEM, CTXm and OXA specific primers by PCR. TEM type was observed in all meat isolates having strong or medium bio film production, followed by CTXm. Antibiotic resistance was found to be more in goat isolates with 68.5%, followed bovine isolates with 56.6% and chicken isolates with 46%. In our study, the strong biofilm producing individual isolates (41%) had at least three of the ESBL genes with higher percentage of antibiotic resistance (>70%) than weak or moderate biofilm producing isolates.
Key words: ß-lactamase, Klebsiella pneumonia
38. Journal of Cell and Tissue Research 14(2): 4409-4412 (2014)
Regeneration of ginger (Zingiber officinale Rosc.) through shoot tip culture
Solanki, R.U.,Parekh, M.J. and Patel, S.R.
Department of Genetics and Plant Breeding, Navsari Agricultural University, Navsari 396450, Gujarat. E. mail: parekh_mithil@yahoo.com,
Abstract: An efficient and systematic protocol for complete plant regeneration via shoot tip culture has been developed for Zingiber officinale Rosc. Shoot tip of ginger treated with HgCl2 at 0.10 per cent concentration performed best results with sterilization. Types of cytokinin and auxin and its concentration influenced the frequency and number of multiple shoots. Profuse multiplication was obtained from shoot tip of young plant on MS medium supplemented with a combination of 6-benzyleaminopurine, naphthalene acetic acid and kinetin. Further plantlets were transferred in half as well as full strength MS medium supplemented with different concentrations of indole 3-butyric acid for effective induction of roots. Regenerated plantlets with well developed roots were transferred for hardening, Cocopeat mixed with clay and FYM gave the maximum survival percentage with better plant growth.
Key words: Zingiber officinale Rosc., Shoot tip culture
39. Journal of Cell and Tissue Research 14(2): 4413-4422 (2014)
Chronic sleep deprivation and its effect on the length of human telomeres: A review
Vartak, S., Barve, S.,Deshpande, A. and Deshpande, P.
Department of Biotechnology, Kelkar Education Trusts’s, V. G. Vaze College, Mulund (E), Mumbai- 400081, Affiliated to University of Mumbai. E. mail: suv21688@gmail.com,
Abstract: Chronic Sleep deprivation has developed into a routine in today’s rapidly growing world. Variety of factors including shift works, medical conditions, sleep disorders, household stress, domestic duties, social and work pressures, etc. are responsible for increased prevalence of Sleep restriction over extended periods and subject body to stress. It produces associated observable physiological and psychological changes. The present analysis has provided certain evidences suggesting that the chronic sleep deprivation as prolonged partial sleep restriction and accelerated telomere attrition are closely linked. Telomeres are the crucial targets for the effects of myriad factors influencing ageing and age related disorders. Telomeres are ‘Biomarkers of Ageing’ and the chronological age advancement is closely associated with these molecular structures. Newly emerging research in telomere biology highlights variety of stress factors with all culminating into accelerated telomere shortening. The present review summarizes the studies that have investigated chronic sleep deprivation as a major stress factor with effects of prolonged sleep loss on telomeres. A brief review of methodologies used to assess sleep and quantify Telomere Length (TL) is done to aid in postulating their feasibility, sensitivity, specificity and reproducibility to help standardize future research directions in this field.
Key words: Chronic Sleep Deprivation, Stress, Ageing, Telomere Length
40. Journal of Cell and Tissue Research 14(2): 4423-4428 (2014)
Pseudomonas and actinobacteria reduce the incidence of leaf curl virus disease in tomato
Gund, S.V., Krishnaraj, P.U.,Jagdeesh, K.S. and Byadgi, A.S.
Department of Biotechnology, University of Agricultural Sciences, Dharwad-580005, India.
E. mail: wishu2810@gmail.com,
Abstract: Fifteen Pseudomonas and five actinomycete isolates were prospected for control of tomato leaf curl viral disease through seed priming, soil and foliar application to tomato plants under green house condition. The inoculation with isolates, AUDP326(4), AUDP360(2), AUDP139, AUDT217 and AUDT152 resulted in significant reduction of disease up to 60-80 per cent. The observation of reduced viral load in rhizobacteria treated plants clearly indicated the suppression of virus multiplication when compared to the disease control. The isolates AUDP360(2), AUDP326(4) and AUDP139 were identified as Pseudomonas fluorescens through 16S rDNA sequence analysis. The reduced tomato leaf curl viral infection and suppression of viral multiplication observed due to inoculation with these isolates indicate its potential utility in the disease control and can form a component of an integrated programme for management of this virus disease in tomato.
Key words: Actinobacteria, Pseudomonas, Tomato leaf curl virus
41. Journal of Cell and Tissue Research 14(2): 4429-4433 (2014)
Gene expression involved in apoptosis- like mechanism before and after freezing of umbilical cord matrix stem cells using different cryoprotectants
Singh, P., Rose, M. K., Yadav, P. S., Sharma, R. and Singh, J.
Department of Veterinary Physiology and Biochemistry, College of Veterinary Sciences,Lala Lajpat Rai
University of Veterinary and Animal Sciences, Hisar 125 004, Haryana. E. mail: raulsikandar@gamil.com,
Abstract: Apoptosis, genetically regulated form of cell death which allows the safe disposal of cells at the point of time when they have fulfilled their intended biological function. An experiment was under taken to study the Gene expression involved in apoptosis –like mechanism before and after freezing of umblical cord matrix stem cells using different cryoprotectants. Umbilical cord tissues from 3 different fetuses were processed and cryopreserved in 3 cryprotectants. Before and after freezing cell viability was assessed using trypan blue dye exclusion method. Cells were cryopreserved in fetal bovine serum and dimethyl sulfoxide combination (FBS-DMSO), ethylene glycol and sucrose combination (EG-Sucrose) and dimethyl sulfoxide trehalose and fetal bovine serum combination (D">.
Key words: Apoptosis, Cryoprotectants
42. Journal of Cell and Tissue Research 14(2): 4435-4441(2014)
Genomic DNA isolation and analysis of genetic variation in Ancistrocladus heyneanus Wall. EX J. graham using ISSR markers
Bhave, R.K., Madan, N.and Barve, S.S.
V. G. Vaze College of Arts, Science and Commerce, Mithagar Road, Mulund (E), Mumbai- 81;
E. mail : raginidange@gmail.com,
Abstract:Ancistrocladus heyneanus Wall ex. J. Graham is an important medicinal plant, endemic to Western Ghats of India, belonging to the family Ancistrocladaceae. The family is unique with respect to the presence of Napthylisoquinoline alkaloids and the biological activity of its constituents, including anti-HIV. In the present research work, the protocol for DNA isolation from leaves has been standardized. The Genetic Diversity between four locations of Ancistrocladus was evaluated using inter simple sequence repeat (ISSR) markers. The average polymorphism detected by selected ISSR primers was 35.95. Genetic parameters consisting of number of effective alleles (Ne), nei genetic diversity (H) and polymorphic information content (PIC) were calculated based on the amplification data. The mean PIC value for selected ISSR markers was 0.4975. The dendrogram obtained from combined molecular data revealed that the population of Hiranyakeshi is distinct from other three, whereas, B1 group of Bhimashankar population is closely related to Matheran population. The ISSR markers proved to be potent tool for diversity analysis studies of Ancistrocladus.
Key words: Ancistrocladus heyneanus, ISSR markers
43. Journal of Cell and Tissue Research 14(2): 4443-4447 (2014)
Antioxidant effect of fenugreek (Trigonella foenum graecum) and turmeric (Curcuma longa) in diet
induced hyperlipidemic rats
Hemanth Gowda, K., Narayana Swamy, M. and Veena, T.
Department of Veterinary Physiology and Biochemistry, Veterinary College, Hassan 573202,
E. mail: hemannagowda@gmail.com,
Abstract: The present study was performed to evaluate the possible antioxidant potentials of Fenugreek seeds and Turmeric rhizome on diet induced hyperlipidemic rats. Hyperlipidemic rat models were created by gavaging the rats with high fat diet made up dalda (vanaspathy) and coconut oil at the dose rate of 10 ml/kg body weight for four weeks. Hyperlipidemic rats were thereafter kept on normal diet and gavaged with Fenugreek seeds (2 to 4 g/kg body weight) and turmeric rhizome (200 to 400 mg/kg body weight) powder for four weeks. Hyperlipidemic rats fed with normal diet were used as positive control. It was observed that treatments significantly inhibited levels of TBARS in hyperlipidemic rats. In addition, the activities of catalase, superoxide dismutase and glutathione peroxidase were augmented after herbal supplement. Histopathological examination of the liver and kidney showed fewer lesions in the hyperlipidemic rats treated with herbs compared with control hyperlipidemic rats. The results suggest that fenugreek seeds and turmeric rhizome would be effective in improving the antioxidant levels.
Key words: Fenugreek, Turmeric, Hyperlipidemia
44. Journal of Cell and Tissue Research 14(2): 4449-4454 (2014)
Molecular analysis of isolates of Fusarium udum causing pigeonpea wilt through RAPD and ISSR
Dhage, S.A., Gupta, V.R., Mane S.S. and Suryawanshi, K.K.
Department of Plant Pathology, PGI, Dr. Panjabrao Deshmukh Krishi Vidyapeeth, Akola, Maharashtra.
E. mail: suryawanshikk@gmail.com, Cell: 0 9421164181
Abstract: The genetic variation was detected among six isolates of Fusarium udum using RAPD and ISSR markers. Out of 30 RAPD primers screened for amplification of DNA of six isolates of Fusarium udum, 18 produced reproducible and scorable bands with 94.8 percent polymorphism. The primer OPA-9, OPB-3 and OPB-14 amplified maximum fragments 12 and the least in OPB-20.. Buldhana isolate had shown higher value of similarity coefficient (0.6428) whereas, Gadchiroli isolate had the lower value of similarity coefficient (0.448). The primer OPA-2, OPA-3, OPA-4, OPA-16 and OPB-17 showed the monomorphic band could be used for identification of Fusarium udum. Similarly, among 16 ISSR markers used, 9 produced reproducible and scorable bands with high percentage of polymorphism. The primer (GA), C amplified maximum fragments (12) and the least in (ATG)6 and (CA)8 RG found (7) fragments. Gadchiroli isolate had higher value of similarity coefficient 0.6125 whereas Akola isolate had the lower value of similarity coefficient (0.40).
Key words: Fusarium udum, Wilt, Pigeonpea
45. Journal of Cell and Tissue Research 14(2): 4455-4461 (2014)
Creation of genetic variability in saffron (Crocus sativus L.) using induced mutation
Salwee, Y. and Nehvi, F.A.
Baba Ghulam Shah Badshah University, Rajouri. Sher-e-Kashmir University of Agricultural Sciences and Technology of Kashmir, Kashmir 191121 J. and K. E. mail: salwees@yahoo.com, Cell: 09419974563
Abstract: Saffron, the world’s famous low volume high value spice growing as a natural heterogeneous temporal subpopulation is a legendary crop of Jammu and Kashmir. Triploid nature of saffron (Crocus sativus L) with genome structure 2n=24 (x=8) is responsible for its sterility. Cytological behavior of saffron eliminates chances of improvement through conventional hybridization methods as commonly practiced in agricultural and horticultural crops for generation of variation with regard to yield and resistance to disease and pests. The chemical mutagen-Ethyl methane sulphonate, Ethidium bromide, colchicine and physical mutagens viz; gamma radiation at different doses have a pronounced effect on floral as well as morphological attributes, viz; number of flowers, fresh flower weight, fresh pistil weight, dry pistil weight, number of leaves, leaf length and leaf thickness. Thus an approach to develop new variants for yield gains using induced mutagens produced yield mutants with increased pistil weight .
Key words: Saffron (Crocus sativus L.), Mutagens, Divergence
46. Journal of Cell and Tissue Research 14(2): 4463-4470 (2014)
In vitro microcorm formation in saffron (Crocus sativus L.)
Salwee, Y.and Nehvi, F.A.
Baba Ghulam Shah Badshah University, Rajouri. Sher-e-Kashmir, University of Agricultural Sciences and Technology of Kashmir, Kashmir 191121. J. and K. E. mail: salwees@yahoo.com,
Abstract: Saffron the sterile triploid belongs to Iridaceae family. Due to triploid nature of saffron sexual reproduction is not possible and it is propagated vegetatively by corms. However the multiplication ratio via corms is very low in nature with a ratio of 1:3 thereby increasing the cost of cultivation and making it the most expensive spice in the world. Tissue culture offers a great potential for commercial corm production in saffron. Present investigation was a step towards standardization of effect of different growth regulators ensuring initial bud sprouting, direct shoot regeneration from the base of spouted buds and cormlet production from multiple shoots for establishment of commercially viable protocol. Initial bud sprouting was achieved using NAA (0.5 mg/l) in combination with BAP (1.5 mg/l) and 2,4-D (2 mg/l) in combination with Zeatin (3mg/l) and BAP (1.5 mg/l).Inoculation of sprouted cultures on MS medium supplemented with 2mg/l Zeatin resulted in shooting Significant shoot proliferation was observed on combining cytokinin with auxins. Shoot multiplicationwas achieved from treatment combination 6mg/l Zeatin and 2mg/l 2,4-D. Minicorms of 8g weight were obtained on growth retardant (CCC@0.2 ml/l) in combination with 9%
Key words: Corm regeneration, Saffron (Crocus sativus L.)
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