ANALYTICAL METHOD DEVELOPMENT, VALIDATION AND STABILITY STUDIES BY RP-HPLC METHOD FOR SIMULTANEOUS ESTIMATION OF ANDROGRAPHOLIDE AND CURCUMIN IN CO-ENCAPSULATED NANOSTRUCTURED LIPID CARRIER DRUG DELIVERY SYSTEM

Objective: The current study aims to boost the bioavailability criteria of two natural bioactive compounds, andrographolide and curcumin by their combination in nanostructured lipid carrier (NLC) and also to develop a straightforward reverse-phase high-performance liquid chromatography (RP-HPLC) method to validate, quantify of andrographolide and curcumin simultaneously in novel NLC formulation. Methods: The reliable chromatographic separation was executed by using a column of Phenomenex octadecylsilane (C18) at 35 °C column oven temperature using a mobile phase of 0.02 M potassium dihydrogen orthophosphate (KH2PO4) salt solution of pH 3.0 as a buffer and acetonitrile in 50: 50 v/v fixed ratio and 1.5 ml/min flow rate of with 20 μl injection load. The detection was carried out at 240 nm isosbestic wavelength employing a photodiode array (PDA) detector. Results: Andrographolide and curcumin were eluted at 2.4 and 4.9 min, respectively. Quantification and linearity were achieved for both drugs at the 10-140 μg/ml range. The method is specified as the presence of excipients utilized in the formulation failed to interfere with the estimation of andrographolide and curcumin. The developed method was successfully utilized to work out the drug loading efficiency and in vitro drug release study of those drugs in NLC formulation and also for the estimation of those drugs from rat plasma. Conclusion: The developed high-performance liquid chromatography (HPLC) method may be utilized in the future estimation of andrographolide and curcumin simultaneously in NLC and other nanoformulations both in vitro and in vivo

Evaluation of Economic Losses due to Coccidiosis in Poultry Industry in India

Coccidiosis is an old parasitic disease, prevalent all over the country and has a significant impact on poultry production. In this paper, economic loss to poultry industry has been estimated considering the major economic parameters. The estimation has revealed that commercial broiler industry is a major sufferer due to coccidiosis wherein 95.61 per cent of the total economic loss occurs due to the disease. The commercial layer industry shares 3.53 per cent economic loss, mainly due to cost of chemoprophylaxis and reduced egg production. A comparison across economic traits has revealed that loss is maximum due to reduced body weight gain, followed by increased FCR (23.74%) and chemoprophylaxis (2.83%) in the total loss due to coccidiosis in broiler industry of India. The overall comparison of economic traits for all the types of poultry sector it has shown that reduced body wt gain and increased FCR are the major parameters from which 68.08 per cent and 22.70 per cent annual loss has occurred in the total loss from coccidiosis in India during the year 2003-04. The total loss due to coccidiosis has been found to be of Rs 1.14 billion (approx) for the year 2003-04. The study has observed that generation of this data across different geographical regions will be helpful to conclude about the global economic loss due to coccidiosis in the poultry industry.Agricultural and Food Policy,

VALIDATION, STABILITY STUDIES, AND SIMULTANEOUS ESTIMATION OF CO-ENCAPSULATED CURCUMIN, EPIGALLOCATECHIN GALLATE NANOFORMULATION BY RP-HPLC METHOD

Objective: A new reverse-phase high-performance liquid chromatography (RP-HPLC) method was developed to simultaneously determine curcumin and epigallocatechin gallate (EGCG) in novel nanoformulation. Methods: The high-performance liquid chromatography (HPLC) method was achieved by using a Thermo Scientific Hypersil Base Deactivated Silica (BDS) C18 column (25 cm X 4.6 mm, 5 µm) at 35 °C column oven temperature. The chromatographic procedure was performed with a mobile phase of acetonitrile and 0.025 M (pH 4.0) potassium dihydrogen phosphate (KH2PO4) buffer by gradient mode of elution. The injection volume was 20 µl, and the flow rate was 1.5 ml/min, with ultraviolet (UV) detection using a diode array detector (DAD) at a 268 nm isosbestic wavelength. Results: Drug entrapment efficiency studies were performed with co-encapsulated EGCG and curcumin nanoformulation, which were found to be 94.35 % and 95.12 %, respectively. This shows that the developed method is highly effective. EGCG and curcumin were eluted at 3.9 min and 10.7 min, respectively. The linearity range was 25-175 µg/ml for EGCG and 12.5-100 µg/ml for curcumin. The correlation coefficient was 0.991 for EGCG and 0.999 for curcumin from the linearity curve, which indicates that the method can produce good sensitivity. Forced degradation studies were conducted in acidic, basic, oxidative, thermal, photolytic, and UV stress conditions, where all the degradation peaks were monitored. Conclusion: The developed method was linear, simple, rapid, robust, and precise. It could be used to quantify EGCG and curcumin simultaneously in various nanoformulations for in vivo and in vitro applications

Intra-species sequence variability in 28s rRNA gene of Oesophagostomum venulosum isolated from goats of West Bengal, India

AbstractObjectiveTo identify genotypes of Oesophagostmum venulosum (O. venulosum) prevailing in West Bengal, India by comparing variation of nucleotide sequences among 28S rRNA.MethodsPCR amplification of partial segment of 28 S rRNA sequence and analysis of sequence amplified product by single strand conformation polymorphism (SSCP).ResultsTwo distinct conformers among male and female parasites were identified by PCR-SSCP analysis. Sequence analysis among conformers revealed the presence of five single nucleotide polymorphisms (SNP) in codon 64, 66, 86, 125 and 146. Secondary RNA prediction structure showed that out of 5 SNPs, 4 occurred at interior loop of RNA which confirmed evolutionary changes among isolates prevailing in this region.ConclusionsSNPs occured in different isolates of O. venulosum might influence critical changes in rRNA folding pattern which influence evolutionary changes among isolates

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Not AvailableGlobally animal models play very important role in the progress of life science in a wide range of biological experimentations. Suitability of different animal model relies on its similarity in genetic makeup, resemble physiological response on diseases of multiple etiology and different drugs, easy maintenance of model animal. Among the fishes, zebrafish and some other species like killifish and cave fish has been found to have great potential and being experimented as an alternative model animal for experimentation of some specific human diseases. Metadata analysis with relevant keywords revealed >8000 research publications on fish model in last 12 years. In India, zebrafish, Danio rerio (Hamilton, 1822), is widely distributed. However, very less attention has so far been paid to understand the importance of the species in biological experimentation except few scattered reports. Besides zebrafish, use of African killifish and cave fish has also been outlined. The present paper synthesized the current knowledge of research and discussed the status and potentials of fish as a model in modern biological experimentation in various medical disorders and recommends their utilization for drug discovery and critical human disease modelling.Not Availabl

Evaluation of Economic Losses due to Coccidiosis in Poultry Industry in India

Coccidiosis is an old parasitic disease, prevalent all over the country and has a significant impact on poultry production. In this paper, economic loss to poultry industry has been estimated considering the major economic parameters. The estimation has revealed that commercial broiler industry is a major sufferer due to coccidiosis wherein 95.61 per cent of the total economic loss occurs due to the disease. The commercial layer industry shares 3.53 per cent economic loss, mainly due to cost of chemoprophylaxis and reduced egg production. A comparison across economic traits has revealed that loss is maximum due to reduced body weight gain, followed by increased FCR (23.74%) and chemoprophylaxis (2.83%) in the total loss due to coccidiosis in broiler industry of India. The overall comparison of economic traits for all the types of poultry sector it has shown that reduced body wt gain and increased FCR are the major parameters from which 68.08 per cent and 22.70 per cent annual loss has occurred in the total loss from coccidiosis in India during the year 2003-04. The total loss due to coccidiosis has been found to be of Rs 1.14 billion (approx) for the year 2003-04. The study has observed that generation of this data across different geographical regions will be helpful to conclude about the global economic loss due to coccidiosis in the poultry industry

Molecular differentiation of cryptic stage of Echinococcus granulosus and Taenia species from faecal and environmental samples

ObjectiveTo differentiate cryptic stage of Echinococcus granulosus (E. granulosus) and Taenia by PCR-RFLP and sequence information of amplicon.MethodsDNA were isolated from metacestodes stage of Taenia and E. granulosus using DNA isolation kit (Q-BIOgene kit, USA), the amplified and purified DNA product was then cloned and sent for sequencing. The generating sequence information was used for amplicons identification.ResultsOut of 112 faecal and environmental samples, 16 exhibited positive result. The product size of amplicon positive for E. granulosus was 310 bp; whereas, for Taenia spp. sizes varied from 379 to 388 bp. Restriction profile of actin II with Csp61 also differed Taenia spp. and E. granulosus.ConclusionsThe result of the study indicated that, the primers were useful to differentiate cryptic stage of the two genera which is yet to be reported earlier

Immunomodulatory Potency of Eclipta alba (Bhringaraj) Leaf Extract in Heteropneustes fossilis against Oomycete Pathogen, Aphanomyces invadans

Aphanomyces invadans is an aquatic oomycete pathogen and the causative agent of epizootic ulcerative syndrome (EUS) in fresh and brackish water fish, which is responsible for severe mortalities and economic losses in aquaculture. Therefore, there is an urgent need to develop anti-infective strategies to control EUS. An Oomycetes, a fungus-like eukaryotic microorganism, and a susceptible species, i.e., Heteropneustes fossilis, are used to establish whether an Eclipta alba leaf extract is effective against the EUS-causing A. invadans. We found that treatment with methanolic leaf extract, at concentrations between 50–100 ppm (T4–T6), protects the H. fossilis fingerlings against A. invadans infection. These optimum concentrations induced anti-stress and antioxidative response in fish, marked by a significant decrease in cortisol and elevated levels of superoxide dismutase (SOD) and catalase (CAT) levels in treated animals, as compared with the controls. We further demonstrated that the A. invadans-protective effect of methanolic leaf extract was caused by its immunomodulatory effect and is linked to the enhanced survival of fingerlings. The analysis of non-specific and specific immune factors confirms that methanolic leaf extract-induced HSP70, HSP90 and IgM levels mediate the survival of H. fossilis fingerlings against A. invadans infection. Taken together, our study provides evidence that the generation of anti-stress and antioxidative responses, as well as humoral immunity, may play a role in protecting H. fossilis fingerlings against A. invadans infection. It is probable that E. alba methanolic leaf extract treatment might become part of a holistic strategy to control EUS in fish species

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Not AvailableThe disease outbreaks in aquaculture system of wetlands are the major cause of fish mortality. Among various bacterial septicaemic diseases, fish mortality caused by Acinetobacter spp. is recently reported in different fish species. Fish disease outbreak was investigated in a wetland of West Bengal, India to identify the aetiological factors involved. The moribund fish were examined and subjected to bacterial isolation. Two bacterial causative agents were identified as Acinetobacter junii and Acinetobacter pittii b y b iochemical c haracterization a nd 1 6S r RNA g ene a mplification. Both the isolates were oxidase-negative, nitrate-negative, catalase-positive and indole-negative. The molecular identification using 16S rRNA gene sequencing and phylogenetic tree analysis further confirmed the two Acinetobacter spp. with 97%– 99% similarity. The antibiotic resistance patterns of these two bacteria revealed that both of them were resistant to β-lactam, cefalexin, cephalothin, amoxyclav, cefuroxime, cefadroxil, clindamycin, vancomycin and penicillin. In addition, A. pittii was also resistant to other antibiotics of cephams group such as ceftazidime and cefotaxime. In the challenge experiment, both A. junii and A. pittii were found to be pathogenic with LD50 of 1.24 × 105 and 1.88 × 107 cfu/fish respectively. Histopathological examination of gill, liver and kidney revealed prominent changes supporting bacterial septicaemia. The investigation reports for the first time on concurrent infection by A. junii and multidrug-resistant (MDR)-A. pittii as emerging fish pathogens to cause severe mortality in Labeo catla and Hypophthalmichthys molitrix in a freshwater wetlandNot Availabl