Alkaline protease from a non-toxigenic mangrove isolate of Vibrio sp. V26 with potential application in animal cell culture

Vibrio sp. V26 isolated from mangrove sediment showed 98 % similarity to 16S rRNA gene of Vibrio cholerae, V. mimicus, V. albensis and uncultured clones of Vibrio. Phenotypically also it resembled both V. cholerae and V. mimicus.Serogrouping, virulence associated gene profiling, hydrophobicity, and adherence pattern clearly pointed towards the non—toxigenic nature of Vibrio sp. V26. Purification and characterization of the enzyme revealed that it was moderately thermoactive, nonhemagglutinating alkaline metalloprotease with a molecular mass of 32 kDa. The application of alkaline protease from Vibrio sp. V26 (APV26) in sub culturing cell lines (HEp-2, HeLa and RTG-2) and dissociation of animal tissue (chick embryo) for primary cell culture were investigated. The time required for dissociation of cells as well as the viable cell yield obtained by while administeringAPV26 and trypsin were compared. Investigations revealed that the alkaline protease of Vibrio sp. V26 has the potential to be used in animal cell culture for subculturing cell lines and dissociation of animal tissue for the development of primary cell cultures, which has not been reported earlier among metalloproteases of Vibrios.

In vivo screening of mangrove plants for anti WSSV activity in Penaeus monodon, and evaluation of Ceriops tagal as a potential source of antiviral molecules

The objective of the study was to find out a natural way to fight white spot syndrome virus (WSSV) in cultured shrimps, as the present scenario necessitated an organic remedy for the devastating pathogen in crustaceans. Under this research programme seven mangrove plants were collected, identified and aqueous extracts screened for their protective effect on the giant tiger shrimp Penaeus monodon against WSSV. The experimental design consisted two modes of application, such as exposure of the virus to the extract and injection challenge, and oral administration of the extract coated feed followed by oral challenge. All experimental animals were monitored through a nested diagnostic PCR analysis. Of the seven mangrove extracts screened aqueous extract from Ceriops tagal imparted total protection to shrimp from WSSV when challenged by both methods. Shrimps administered with the aqueous extract from C. tagal were devoid of virions. The HPLC fingerprint of the aqueous extracts from C. tagal showed more than 25 peaks and 7 of them were larger and well separated. Preliminary phytochemical analysis revealed the presence of alkaloids, flavonoids, polyphenolics, cardiac glycosides, saponins and sterols. The study indicated suitability of the aqueous extract of C. tagal as a possible prophylaxis for WSSV infection in shrimp. This is the first report on the anti WSSV property of the mangrove plant C. tagal

Characterization and applications of two protease enzymes obtained by culture dependent and independent approaches from mangrove sediments

Soil community genomics or metagenomics is employed in this study to analyze the evolutionary related - ness of mangrove microbial community. The metagenomic DNA was isolated from mangrove sediment and 16SrDNA was amplified using universal primers. The amplicons were ligated into pTZ57R/T cloning vector and transformed onto E. coli JM109 host cells. The recombinant plasmids were isolated from positive clones and the insert was confirmed by its reamplification. The amplicons were subjected to Amplified Ribosomal DNA Restriction Analysis (ARDRA) using three different tetra cutter restriction enzymes namely Sau3A1, Hha1 and HpaII. The 16SrDNA insert were sequenced and their identity was determined. The sequences were submitted to NCBI database and accession numbers obtained. The phylo - genetic tree was constructed based on Neighbor-Joining technique. Clones belonged to two major phyla of the bacterial domain, namely Firmicutes and Proteobacteria, with members of Firmicutes predominating. The microbial diversity of the mangrove sediment was explored in this manner.

Community Participation in Ecotourism: An Inclusive Development Option for Kerala

The inferences obtained from the study are presented in coherent area-specific levels so as to understand the ecotourism and its sub-sector areas for the researchers and policy makers about the issues, importances and potentialities of the sector. An analysis of the tourism sector in Kerala has shown tremendous growth both in terms of tourist arrivals and in terms of revenue generation from direct and indirect sources. The foreign tourist visitors in Kerala in 2014 was 9,23,336 which shows 7.60 percent increase from the last year and the domestic tourist visitors were 1,16,95,411 which again shows 7.71 percent increase, is a clear evidence of its potential. In 2014 the industry contributed revenue of 24885.44 crores from direct and indirect sources giving rise to an increase of 12.11 percent from the last year. A dichotomy of tourists and ecotourists shows that tourists in the ecotourism destinations come to 42.6 percent of the total, shows the scope, significance and its potential. Correlation of zone-wise tourist arrivals based on the ecotourism destinations highlights the fact that with only 19 of the 64 destinations that come in the central zone are the most preferred centres (around 54 percent) for the domestic as well as foreign tourists. The north zone encompassing 6 districts with rich biodiversity shows that the tourists‟ arrival patterns exhibit less promising results. Though the north zone has 31 ecotourism destinations of the state receives only 6.19 percent of the foreign visitors. The ecotourism activities in the state are primarily managed by the Eco-Development Committees (EDCs) and the Vana Samrakshana Samithies (VSS) under the Forest Development Agency of Kerala. Social class-wise categorization of membership shows that 13142 families have membership in 190 EDCs with SC (28 percent), ST (33 percent) and other marginalised communities (39 percent). But this in the VSS shows that 400 VSS have 59085 members actively engaged in ecotourism activities and social category of the VSS makes clear that majority are from the other marginalized fringe households with 62 percent where as the participation of SC is 12 percent and ST is 26 percent. An evaluation of the socio-economic and demographic matrix of the community members involved in ecotourism activities brings out region specific differences. About 75.70 percent of the respondents are males and the rest are females. Majority of the respondents (about 60 percent) are in the age group of 20 to 40 years, followed by the age group of 40-50 (20 percent). The average age of respondents in the three zones is between 35 and 37 years. The majority of the respondents are married, a few are unmarried. Average family size is 4-5 members and differences are identified among zones. Average number of adults per household is 3 and child per household is 2. Majority have an education of 10th class and below i.e. about 60 percent of the sample have only basic school education like primary, secondary and high school (i.e. up to SSLC but not passed) level. About 18 percent are SSLC passed, 10 percent are undergraduates whereas 6 percent constitute respondents having qualification of graduation and above. Majority of the „graduates and above‟ are from south and central zone. Inter-zone differences in educational profile are also identified with lesser number of „graduates and above‟ are identified in the north zone compared to the other two zones. Investigating into the income and livelihood options of the respondents gives insight about the prominence of ecotourism as an employment and livelihood option for the community members, as more than 90 percent of the respondents have cited tourism sector as their main employment option. Most (49.30 percent) of respondents get 100 percent income from tourism related activities, followed by 37.30 percent of community members have income between 75-99 percent from tourism whereas the rest (13 percent) have less than 74 percent of their income from tourism and there exists difference between zones and percentage of income. Financial habit shows that about 49.7 percent hold active bank accounts, 61 percent have savings behaviour and 73.8 percent have indebtedness. Analysis about the ownership of house brings to light that 37 percent of respondents live in their own house followed by 25.7 percent in government funded/provided house and 21 percent in their parent‟s house and 3.5 percent in rented house. About 12 percent of the respondents have other kinds of accommodation facilities such as staff quarters, etc. But in the case of north zone majority i.e. 52 percent primarily depend on the government funded house indicating the effectiveness of government housing programme. Standard of living measured in SLI frameworks shows that majority of the respondents have medium SLI values (42.3 percent); the remaining 47.7 percent have low SLI and 10 percent have high SLI. The community members have been benefitted immensely from forest and its resources. Since the ecotourism destinations are located amidst the wildlife settings, majority of them depend on forest for their livelihood. The information on the tourist‟s demographic characteristics like age, sex, educational qualification and annual income show that the age category of domestic and foreign tourists falls below the age group of less than 35 years (about 65 percent), whereas only 16 percent of tourists are aged above 46 years. The age group below 25 years consists of more international tourists (31.3 percent) compared to the proportion of domestic tourists (12.5 percent). Male-female ratio shows that the males constitute 56 percent of the sample and females with 44 percent. The factors determining the impact of ecotourism programmes in the community was evaluated with the aid of a factor analysis with 12 selected statements. The worries and concerns of the community members about the impact of ecotourism on the environment are well understood from this analysis. It can be drawn that environment protection and the role of ecotourism in improving the income and livelihood options of the local communities is the most important factor concerning the community members.