DEVELOPMENT OF HIGH-RESOLUTION MOLECULAR TECHNOLOGIES TO INVESTIGATE ECOSYSTEM CHANGES IN THE SOUTHERN EUROPEAN SEAS: THE PROTOTYPING OF DNA CHIPS TO IDENTIFY INVERTEBRATE PREY SPECIES IN MARINE FOOD WEBS OF ADRIATIC SEA

A invertebrate DNA microarray targeting ten species common in the Mediterranean was developed within the framework of the FP6 STREP FISH& CHIPS. Based on sequence variation of the 16S rDNA gene, we designed species-specific probes that were used to construct a DNA chip prototype for the identification of invertebrate (i.e. molluscans and crustaceans) preys of demersal fishes in the Adriatic Sea. The constructed invertebrate DNA chip represents a technological advance towards the wide use of rapid and high-resolution molecular tools to assess structure and changes of marine ecosystems such as the monitoring of invasive species and of re-emerging pathogens in ballast waters

Invertebrate DNA Chip: Opportunities and Challenges in the Development and Application of Microarrays for Marine Biodiversity Studies

DNA microarrays for marine biodiversity studies have been developed and tested in several groups, spanning different applications. They are employed to characterize ecological communities circumventing uncertainties and challenges associated with the conventional techniques of taxonomy employed to characterize these communities. Examples come from identifications of fishes and phytoplankton to monitoring of harmful algae. Although DNA microchips support a great diversity of applications and provide a wealth of findings in functional genomics and environmental studies, limits in ecological applications are known. So far, DNA microarrays used in biodiversity studies provide only qualitative data in terms of presence or absence of the species and its usage is greatly limited in providing the quantitative measurement of environmental samples. Moreover, DNA microarrays are commonly restricted to identify only those species that are targeted by the probes implemented on the chip. In turn, developing a broad spectrum array has technological challenges in terms of probe designing, experimental optimization, and statistical analysis. In this book chapter, we provide a critical assessment on promises and pitfalls of DNA microarrays as tool for marine invertebrate species identification. We implemented a DNA-chip prototype to identify 15 species of marine invertebrates from European Seas, including crustaceans, molluscs, and polychaetes, based on the two mitochondrial markers, cytochrome oxidase subunit I and 16S rRNA. Challenges involved in oligonucleotide probe design, in silico evaluation, and difficulties encountered through hybridization experiments are here explored. Specificity and sensitivity of the probes have also been critically evaluated to verify the suitability of the selected markers for microarray probe design. Since ultimate application of DNA microarray to resolve complex environmental samples is a major challenge, we made an attempt to analyse gut contents of predator fishes. The problems encountered during this analysis, as the presence of target and not target species that could affect the specificity and the sensitivity of the DNA-chip to distinguish low and high abundant target species from a background of non-targets, were explored. We also reviewed advantages and disadvantages of DNA microarray technology compared to other molecular identification methods that recently spread, i.e. DNA barcoding and next generation sequencing. The outcome of the Invertebrate DNA Chip prototype served as a proof-of-concept for the identification of selected marine invertebrates and prey species of demersal fishes by DNA microarray. The potential of such microarrays can encompass several fields of scientific applications in marine biodiversity and ecosystem sciences, as marine ecosystem diversity and environmental monitoring, seafood quality control, and understanding food webs and ecosystem functioning

Knowledge, awareness, and behavior study on HIV/AIDS among engineering students in and around Guntur, South India

Background: Human Immunodeficiency Virus (HIV) is one of the important contagion issues in the world. It poses serious challenges not only to health professionals but professionals across industries. In India, in general, studies on HIV are carried out among medical and paramedical students and least among non- medical students, whose awareness is of prime importance. Aim: This study was designed to assess the knowledge, attitude, and behavior regarding HIV/AIDS among engineering students in and around Guntur, Andhra Pradesh. Patients and Methods: A cross-sectional study was conducted among 400 engineering college students in and around Guntur. A self-administered questionnaire was given and the results were tabulated and analyzed. Statistical Analysis Used: The data was entered in excel sheet and analyzed using SPSS v. 20.0 software. Data was analyzed using student's t-test and Chi-square test. Results: Out of 400 students, 257 (64.2%) were males and 143 (35.7%) were females. Maximum students (97.2%) indicated they know about HIV/AIDS. Eighty-nine percent (89.7%) of the students responded that needle prick injury can transmit HIV infection, whereas 82.5% of the responders knew that HIV/AIDS affects immune systems. A total of 66.5% of individuals agreed that there is no cure for HIV/AIDS and 72.0% of individuals responded that HIV/AIDS cannot be transmitted through saliva and 20.5% of the students felt that it is necessary to isolate the infected individuals from general public. Conclusions: In addition to medical and para medical students, it is very important for the youth to be aware of HIV/AIDS and its social implications

Methods for Seafood Authenticity Testing in Europe

56 pages, 5 figuresSeafood authenticity is a key parameter for seafood quality, particularly in Europe where regulations provide a strict framework for seafood labeling. A wide variety of methods are commonly used in control laboratories (private or public) to identify seafood species, but emergent approaches for the development of new and fast DNA- and protein-based methods for species differentiation are also considered. To address the challenges in controlling further labeling requirements in the latest European legislation on seafood product traceability and labeling (Regulation (EU) 1379/2013), a review of the development of methods to identify fishing areas and to distinguish between wild and farmed fish, as well as an overview of the advanced methods that could be used for differentiation of fresh and frozen-thawed fish, is given. These methods will become increasingly important in the near future as the risk-based control of food authenticity is prescribed by the new EU control regulation (Regulation (EU) 2017/625)N