Absract Archive May 2008 Research article Optimization and Characterization of Bluetongue Virus in Embryonated Chicken Egg Abstract Bluetongue (BT) is an infectious, non contagious arthropod-borne disease of ruminants caused by Bluetongue virus (BTV), prototype species of the genus Orbivirus, within the family Reoviridae. Bluetongue outbreak was studied in detail in six western districts of Tamilnadu namely, Namakkal, Salem, Erode, Coimbatore, Karur and Dindigul during 2003-2006. The classical signs of cyanosis of the tongue and reddening of the coronary band are a common feature of the disease in native sheep, Macheri breed were observed. 165 blood samples were collected from the field outbreak (95 numbers) have been used to demonstrate BTV and further inoculated in embryonated chicken eggs (11 - 13 day old). In ECE inoculation technique, 13 positive results were obtained in a total of 165 samples (7.87 % positive). The 13 BTV positive samples further inoculated in ECE in two different routes namely, yolk sac route and intravenous route for studying infectivity titres of Bluetongue virus upto 12 passages and compared. High infectivity titre value, i.e., 6.49 and 7.13 was observed in 12th passage in yolk sac and intravenous route of inoculation which was confirmed by higher mean difference (0.642 p=0.019). Intravenous route of inoculation exhibited better infectivity titre value than yolk sac. The thirteen isolated BTV isolates were acid labile, stable at 37oC and resistant to ether, chloroform and RNAse. Key Words: Bluetongue Virus, Embryonated Chicken Egg, EID50, Introduction Bluetongue (BT) is an infectious, non-contagious arthropod-borne disease of ruminants caused by Bluetongue virus (BTV), prototype species of the genus Orbivirus, within the family Reoviridae. Twenty four serotypes of bluetongue virus have been identified till date (Davies et al., 1992). India has significant population of domestic and wild ruminants, which are known to be susceptible to bluetongue virus infection and total sheep population is 51 million, accounting to 5 % of world's sheep population and 123 million goats accounting for 20 % of the total global livestock (FAO, 2003). Hence bluetongue has become one of the important sheep diseases of the Indian subcontinent. The disease was first reported in India in 1964 (Sapre, 1964). Bluetongue in India is endemic in Tamil Nadu, Andhra Pradesh, Karnataka, Maharashtra, Gujarat, Rajasthan, Haryana, Himachal Pradesh, Jammu and Kashmir. However the recent epidemics in Tamil Nadu were devastating (Wilson et al., 1997). India earns Rs.8500 crores annually through the production of meat, wool and skin from 485 million livestock, of which Tamil Nadu accounts for 24 million livestock (Seventeenth Livestock Census, 2003). Small and medium farmers in Tamil Nadu rear sheep and goats for meat and skin, which fetch Rs.200 for an individual animal, and the current total number of sheep and goats for meat and skin records 5.2 and 6.4 million respectively. The outbreak of the bluetongue disease in Tamil Nadu and its occurrence in many parts of India over the last few decades have affected millions of sheep, goats and other livestocks (Ilango, 2006). Sustained growth of the sheep and goat industry depends on eradication of such emerging disease. In Tamil Nadu, 22 out of 24 districts were reported to be affected by the bluetongue virus. Authors:Sekar.P, Gurusubramanian.G,Ponmurugan. K. Research article An approach to suggest the possibilities of interspecific hybridization using RAPD analysis of some commercial fruits and vegetables for improving the genetic characteristics. Abstract RAPD (Random Amplified Polymorphic DNA) analysis is a PCR based molecular marker technique. Here, single short oligo nucleotide primers are arbitrarily selected to amplify a set of DNA segments distributed randomly through out the genome. RAPD was the first to become available and is most commonly and frequently used. Vegetables provide many essential vitamins and minerals. Additionally, higher intakes of vegetables are associated with healthier lives including lower risks of cancer and coronary heart disease. It is recommended that children eat five fruits and vegetables a day. Here different species and varieties of vegetables are used to know the diversity between them. The DNA is isolated and subjected for PCR amplification .The PCR done samples are run on agarose electrophoresis .The analysis is carried out using RAPD software. Key words: RAPD, PCR, primers, Amplification, agarose electrophoresis. Introduction Humans evolved from ancestral apes, and our genes are still reflecting a vegetation and small animal eating past. Evolving in Africa and then Asia, we ate a huge range of leaves, buds, flower buds, stems, gums, roots, tubers, and even pollen. The number of plant families we used as food was very much greater than the restricted range we eat today. Wild foods were carefully selected to avoid the plants or parts of plants with bitter and unpleasant taste, which likely contained toxic compounds. Today's plants are more palatable, and yet paradoxically, we eat very few plants as part of our daily diet. Author:B. Bharathiraja. Review Borrelidin: A promising anticancer agent from Streptomyces species. Abstract Borrelidin, a novel nitrile containing macrolide antibiotic obtained from the Streptomyces species particularly inhibits threonyl-tRNA synthetase (THrRS). Also borrelidin was found to possess antiangiogenic activity- a key process in the spread of malignant tumors by inducing the collapse of formed capillary tubes. Biologically active analogues of borrelidin can be produced by incubating the strains with alternative starting materials. Borrelidin analogues were also found to be the cytotoxic against some cancer cell lines. This review provides the analysis of borrelidin and its analogues from bacteria. Keywords: borrelidin, threonyl-tRNA synthetase, polyketide synthase, angiogenesis. Introduction Borrelidin, 2-(7-cyano-8, 16 dihydroxy-9, 11, 13, 15-tetramethyl-18-oxooxa cyclo octadeca 4, 6-dien-2yl) cyclo pentanecarboxylic acid is first isolated from Streptomyces rochei in 1949. It is crystalline white solid in appearance with molecular weight of 489.6 (molecular formula - C28H43NO6), which is soluble in organic solvents like Dimethyl Sulfoxide (DMSO) and Ethanol. Borrelidin, 18 membered polyketide macrolide antibiotic, is structurally unique and rarely occurs in nature.1 The planar structure of borrelidin was elucidated by Keller-Schierlein in 19672 and subsequently refined by the Nuclear Magnetic Resonance (NMR) analysis and X-ray diffraction methods3,4. The absolute configuration of its nine stereogenic centers has been determined by the X-ray diffraction of its crystal in chiral solvent.4 The unique structural features of borrelidin includes (i) a deoxy propionate subunit consisting of four 1,3 alternating C-methyl groups with a distinctive syn/syn/anti relationship at C4 - C10 (ii) a conjugated diene nitrile chromophere unit at C12 - C15 and (iii) a cyclo pentane carboxylic acid subunit at C17. (Figure 1) The nitrile, lactone and probably the hydroxyl functions are essential for the borrelidin molecule to show antimicrobial activity. The structural features of borrelidin were similar to the antibiotic, Treponemycin.5, 6,7 Biosynthesis of borrelidin Although, Borrelidin first isolated from a soil sample of Streptomyces rochei subsequently identified from other Streptomyces species such as S. parvulus, S. albovinaceous, S. Griseus and an unidentified Streptomyces species C2989.8-12 Apart from microbial sources, borrelidin also can be synthesized chemically which has similar structure and functions. Novel analogues of borrelidin also can be synthesized by precursor directed biosynthesis. 5,13 Author:Vino S. Mini Review Stress Responses- Unique Survival Stratagies of Bacteria Abstract Microbes being the smallest living creatures are no exception when it comes to stress responses. Different micro organisms show varieties of stress responsive mechanisms which in turn help them to survive even under adverse situations like sudden increase/decrease in temperature, lack of nutrient availability and so on. In case of extremophiles, they prefer to live in stressful conditions rather than that of natural environments. It is wonderful to have a deep insight into these unique mechanisms of stress responses in microbes. These mechanisms not only act to provide protection to the microbes but theses are remarkable adaptations showing natural equipments provided by nature to the micro organisms through evolution. Key word : Microbial Stress Response, Extremophiles, Probiotics, Bioremediation. Introduction The word stress has become a very frequently encountered word in day to day life for human beings. But even for a microscopic organism like bacteria there is not much a change in scenario. An organism's survival from moment to moment depends, at least in part, on its ability to sense and respond to changes in its environment. Mechanisms for responding to environmental changes are universally present in all living beings. Bacteriaand especially those capable of persisting in diverse environments, provide particularly valuable models for exploring how single-celled organisms respond to environmental stresses. Bacteria must sense the changes and then respond with appropriate alterations in gene expression and protein levels. Hence it can be said that different stress responses present in different bacteria and their regulations do play significant roles in their survivals especially in adverse conditions. What is stress response in bacteria? It is difficult to provide a universal definition for stress since the perception of a stressful situation is highly dependent on an individual cell. Keeping this in mind, stress can be defined as ' ANY DEVIATION FROM OPTIMAL GROWTH CONDITIONS THAT RESULTS IN A REDUCED GROWTH RATE'. There are different levels of stress severity. Under minor stress, growth usually continues at the same rate and cells fully adapt to the new conditions. Under severe stress, the growth rate gets reduced but cells can still tolerate the state. Under extreme stress, growth ceases and devotes their resources to survival. Under lethal condition, responses may lead to the sacrifice of many cells. Author:Indrani Bhattacharyya. Short communication Resistance in E.coli to antibiotics Abstract The major problems facing today are that many of the disease causing bacteria have become resistant to the effects of different antibiotics. E.coli is one of the most studied bacteria. The E.coli MTCC No.443 was revived and cultured in nutrient broth. The antibiotic sensitivity tests were performed to all the antibiotics such as Ofloxacin, Norfloxacin, Ampicloxacillin, Amoxicillin, Ampicillin, Penicillin, Metronidazole, and Erythromycin. It was found that that the Standard organism showed sensitivity to Ofloxacin, Norfloxacin, and Erythromycin. The E.coli was isolated from an infected patient sample and was confirmed by the biochemical tests. The antibacterial activity tests were done with the same antibiotics mentioned above and showed that the organism showed resistance to all the antibiotics. The Plasmid DNA of the E. coli Sample was isolated and was run on the Agarose gel electrophoresis, and assumption was made that the plasmid DNA could be of a larger molecular weight Introduction Bacteria exists nearly every surface and within a variety of living and non-living things. They exist in a symbiotic condition in which they function as partners with other organisms. Although life would not be the same without bacteria, they also have a bad side. These bad or pathogenic bacteria cause diseases and other unwanted infections. To fight these bacteria we use antibiotics; a natural defense used by other organisms that humans have adopted. E.coli is one of the most studied bacteria. E.coli are essential for producing vitamins K and B complex. Our bodies are dependent on E.coli for the production of these vitamins, our only source. Although we depend on these helpful strains of E.coli there are some harmful strains of E.coli like the 0157:47 strain that have been associated with a wide variety of diseases and infections, including menigeal predominantly in the newborn gastro intestinal, urinary tract, wound and bacterium infections in all age groups. These categories of diarrheogenic E.coli also cause numerous types of diarrhea illness. One of the major problems facing today is that many of the disease causing bacteria have become resistant to the effects of different antibiotics. This occurs when a bacteria population genetically acquires the ability to destroy the antibiotics. This occurs when a bacteria population genetically acquires the ability to destroy the antibiotic. This one resistant cell will divide and produce a population that is now no longer harmed by that particular antibiotic. This is a great concern because certain strains of disease causing bacteria now have only few antibiotics remaining, which will kill them. Due to this concern there are tremendous efforts in finding new natural sources of antibiotics and or make completely synthetic ones in the laboratory. One of the reasons for the cause of this problem is because of prior indiscriminate use of antibiotics in humans. Authors:Karuna Jyothi, Priya.R.Iyer. Tools & Techniques From Ideogram to Spectrogram Application of FISH Technique in clinical diagnosis The most recent developments in cytogenetics have led to the "Molecular Era". Advances in the use of DNA probes have allowed cytogeneticists to hybridize these probes to chromosomes and determine if a specific DNA sequence is present on the target chromosome. This has been useful in detecting anomalies beyond the resolution level of conventional banded chromosomes and also in determining the location of specific genes on chromosomes. Fluorescence in situ hybridization (FISH) as mentioned is used as an additional staining method for metaphase as well as interphase chromosome analysis. During the past 5 years, FISH has begun to demonstrate its considerable diagnostic potential in cancer, prenatal and micro-deletion syndromes. 1. FISH and cancer cytogenetics The study of acquired chromosomal anomalies in cancer continues to expand. Cancer cells are often difficult to grow in culture and FISH techniques can augment standard cytogenetic testing. Numeric changes such as trisomy 8 in myeloid disorders can be detected. Structural chromosomal aberrations - translocations, such as the bcr/abl rearrangement in chronic myelogenous leukemia and acute lymphoblastic leukemia can also be easily identified. FISH studies are being used to look for early relapse and residual disease in nondividing cells. In allogeneic bone marrow transplant patients receiving opposite sex donor cells, the success of engraftment can be monitored by FISH chimerism studies. Differentially labeled X- and Y-specific probes can be used to detect the proportions of XX to XY cells in bone marrow or peripheral blood nuclei in a dual color FISH procedure. By combining immunocytochemical detection of cancer cells and FISH, chromosomal abnormalities and cell types can be simultaneously studied. Gene amplification like Neu and Her-2 also can be detected in solid tumour sections as well as cultured cells. Authors:Solomon F.D. Paul, Vijayalakshmi J, Koshy T, Kaur H, Venketswaran N. & Venkatachalam P.