Microsatellite Genotyping Corroborated Loss of Genetic Diversity in Clarias batrachus as a Result of Lack of Regulatory Reforms in Aquaculture.

In India, over the past 50 years, aquaculture practices of species such as those used for Clarias batrachus were developed without adequate regulatory oversight. In these situations, it is important to consider the influence that genetic factors can have on such vulnerable aquaculture species. Population genetic structure can be evaluated through the use of neutral molecular markers, and this can aid in predicting the risk of the demise of populations and for framing management strategies to conserve remaining populations. The study presented here reports on the genetic status of C. batrachus populations through the analysis of data collected using 22 microsatellite markers from seven natural and one hatchery population. The mean values for observed heterozygosity across loci within populations ranged from 0.242 to 0.485. Measures of genetic differentiation were low overall, with mean values for FST of 0.270, FIS of 0.113 and FIT of 0.353. An AMOVA analysis revealed that percentages of variation among and within populations were 27.16 and 6.86, respectively, and Bayesian clustering analyses showed a population subdivision consisting of five clusters with admixture of haplotypes from other populations leading to genetic bottleneck. We also examined how hatchery management factors leading to excessive exchanges of fish between river systems through could impact the structure of the C. batrachus populations. Overall, this study shows how the systematic use of molecular markers can facilitate the development of management policies for these populations and for the development of a comprehensive set of rules for hatcheries and aquaculture practices, including avoidance of excessive homozygosity by avoiding repeated use of feral broodstock and their interrogation.

Multiple tests on saffron find new adulterant materials and reveal that Ist grade saffron is rare in the market.

Among spices, Saffron is among the most extensively interrogated for purity and authenticity. Numerous methods have been recommended for authentication of Saffron samples and for detection of adulterants for codex compliance. However, none of these methods can fulfill both of these important quality criteria. This study describes a three step approach to achieving this goal by including the established ISO3632 method and two additional methods based on microscopic examination and DNA barcoding. We provide results showing the utility of these methods both independently and in combination for quality evaluation of 36 commercial saffron samples. Our results show that use of the ISO3632 approach alone can reveal the color and aroma but not the genetic origin of the material or distinguish between synthetic components versus natural ingredients. Also, the microscopic observation method can give a preliminary indication of saffron authenticity, but used alone it is unable to quantify purity. Finally, a relatively new method based on the use of DNA barcodes can authenticate the biological origin of the saffron, but here results may be misleading if auto-adulterating materials are present. Overall, our study reveals that through the combined use of all three methods, saffron authentication can substantially improved.

DNA barcode based wildlife forensics for resolving the origin of claw samples using a novel primer cocktail.

Excessive wildlife hunting for commercial purposes can have negative impacts on biodiversity and may result in species extinction. To ensure compliance with legal statutes, forensic identification approaches relying on molecular markers may be used to identify the species of origin of animal material from hairs, claw, blood, bone, or meat. Using this approach, DNA sequences from the COI "barcoding" gene have been used to identify material from a number of domesticated animal species. However, many wild species of carnivores still present great challenges in generating COI barcodes using standard "universal" primer pairs. In the work presented here, the mitochondrial COI gene was successfully amplified using a novel primer cocktail, and the products were sequenced to determine the species of twenty one unknown samples of claw material collected as part of forensic wildlife case investigations. Sixteen of the unknown samples were recognized to have originated from either Panthera leo or P. pardus individuals. The remaining five samples could be identified only to the family level due to the absence of reference animal sequences. This is the first report on the use of COI sequences for the identification of P. pardus and P. leo from claw samples as part of forensic investigations in India. The study also highlights the need for adequate reference material to aid in the resolution of suspected cases of illegal wildlife harvesting.

Genetic structure of populations and conservation issues relating to an endangered catfish, Clarias batrachus, in India.

The Asian catfish, Clarias batrachus (Linnaeus, 1758), is a highly valued species endemic to India that is currently in drastic decline in most of its natural habitat. The present study was undertaken to document the genetic structure of populations of this species using mitochondrial DNA markers, specifically from the cytochrome B and D-loop regions. Specimens from eight wild populations were collected and analyzed from different regions in India. The genetic variation within and among populations was evaluated using a range of descriptive statistics. The analysis described here provides a broad and consistent view of population structure and demographic history of populations of C. batrachus. Although there was some genetic structuring consistent with regional differences, all eight populations examined here showed relatively low levels of genetic variation in terms of both haplotype and nucleotide diversities in the different analyses used. However, a number of private haplotypes were discovered, and this may provide valuable information for future selective breeding program and conservation management. The results may aid in the design and implementation of strategies for the future management of this endangered catfish C. batrachus in India.