Saline-saturated DMSO-EDTA as a storage medium for microbial DNA analysis from coral mucus swab samples

The mucus surface layer of corals plays a number of integral roles in their overall health and fitness. This mucopolysaccharide coating serves as vehicle to capture food, a protective barrier against physical invasions and trauma, and serves as a medium to host a community of microorganisms distinct from the surrounding seawater. In healthy corals the associated microbial communities are known to provide antibiotics that contribute to the coral’s innate immunity and function metabolic activities such as biogeochemical cycling. Culture-dependent (Ducklow and Mitchell, 1979; Ritchie, 2006) and culture-independent methods (Rohwer, et al., 2001; Rohwer et al., 2002; Sekar et al., 2006; Hansson et al., 2009; Kellogg et al., 2009) have shown that coral mucus-associated microbial communities can change with changes in the environment and health condition of the coral. These changes may suggest that changes in the microbial associates not only reflect health status but also may assist corals in acclimating to changing environmental conditions. With the increasing availability of molecular biology tools, culture-independent methods are being used more frequently for evaluating the health of the animal host. Although culture-independent methods are able to provide more in-depth insights into the constituents of the coral surface mucus layer’s microbial community, their reliability and reproducibility rely on the initial sample collection maintaining sample integrity. In general, a sample of mucus is collected from a coral colony, either by sterile syringe or swab method (Woodley, et al., 2008), and immediately placed in a cryovial. In the case of a syringe sample, the mucus is decanted into the cryovial and the sealed tube is immediately flash-frozen in a liquid nitrogen vapor shipper (a.k.a., dry shipper). Swabs with mucus are placed in a cryovial, and the end of the swab is broken off before sealing and placing the vial in the dry shipper. The samples are then sent to a laboratory for analysis. After the initial collection and preservation of the sample, the duration of the sample voyage to a recipient laboratory is often another critical part of the sampling process, as unanticipated delays may exceed the length of time a dry shipper can remain cold, or mishandling of the shipper can cause it to exhaust prematurely. In remote areas, service by international shipping companies may be non-existent, which requires the use of an alternative preservation medium. Other methods for preserving environmental samples for microbial DNA analysis include drying on various matrices (DNA cards, swabs), or placing samples in liquid preservatives (e.g., chloroform/phenol/isoamyl alcohol, TRIzol reagent, ethanol). These methodologies eliminate the need for cold storage, however, they add expense and permitting requirements for hazardous liquid components, and the retrieval of intact microbial DNA often can be inconsistent (Dawson, et al., 1998; Rissanen et al., 2010). A method to preserve coral mucus samples without cold storage or use of hazardous solvents, while maintaining microbial DNA integrity, would be an invaluable tool for coral biologists, especially those in remote areas. Saline-saturated dimethylsulfoxide-ethylenediaminetetraacetic acid (20% DMSO-0.25M EDTA, pH 8.0), or SSDE, is a solution that has been reported to be a means of storing tissue of marine invertebrates at ambient temperatures without significant loss of nucleic acid integrity (Dawson et al., 1998, Concepcion et al., 2007). While this methodology would be a facile and inexpensive way to transport coral tissue samples, it is unclear whether the coral microbiota DNA would be adversely affected by this storage medium either by degradation of the DNA, or a bias in the DNA recovered during the extraction process created by variations in extraction efficiencies among the various community members. Tests to determine the efficacy of SSDE as an ambient temperature storage medium for coral mucus samples are presented here.

Factors influencing colour production from the sugar medium used for the rapid approximation of bacterial counts in fishery products

Results of the studies carried out to elucidate the factors influencing colour production from the sugar medium used for the rapid approximation of bacterial counts in fishery products are reported. The effect of particle size, trace elements, salt soluble protein and non-protein fractions, rate of multiplication of bacteria, in the medium, surface bacteria and the rate of colour production by individual strains of bacteria were studied. It is observed that the best results are obtained when a sea-water homogenate is used.

CFD-DEM simulation of propagation of sound waves in fluid particles fluidised medium

In this work, speed of sound in 2 phase mixture has been explored using CFD-DEM (Computational Fluid Dynamcis - Discrete Element Modelling). In this method volume averaged Navier Stokes, continuity and energy equations are solved for fluid. Particles are simulated as individual entities; their behaviour is captured by Newton's laws of motion and classical contact mechanics. Particle-fluid interaction is captured using drag laws given in literature.The speed of sound in a medium depends on physical properties. It has been found experimentally that speed of sound drops significantly in 2 phase mixture of fluidised particles because of its increased density relative to gas while maintaining its compressibility. Due to the high rate of heat transfer within 2 phase medium as given in Roy et al. (1990), it has been assumed that the fluidised gas-particle medium is isothermal.The similar phenomenon has been tried to be captured using CFD-DEM numerical simulation. The disturbance is introduced and fundamental frequency in the medium is noted to measure the speed of sound for e.g. organ pipe. It has been found that speed of sound is in agreement with the relationship given in Roy et al. (1990). Their assumption that the system is isothermal also appears to be valid.

Evolution of suitable trawl nets for medium size trawlers - 1: Comparative fishing efficiency between 32 m bulged belly, long wing and four panel trawls

Comparative studies of the efficiency of 32 m bulged belly, long wing and four panel trawls have shown that the bulged belly trawl to be superior to the other nets in catching bottom fishes and column fishes. 40% of the bottom fishes and 48% of the column fishes were caught by the bulged belly trawl. However, for prawn catch, the long wing trawl appears to be better as it landed 52% of the total prawn catch of the three nets. Bulged belly trawl was found to be next only to long wing trawl in this respect.

Development of trawls for medium sized trawlers for Veraval, north-west coast of India

Comparative fishing experiments with 25 m bulged belly and 25 m six seam trawls were carried out to study the relative efficiency of the gear. Bulged belly trawl was found more efficient than the other at depths below 40 m. The tension and horizontal opening were more in bulged belly and six seam trawl respectively. Bulged belly caught more of prawns and lobsters but there was no significant difference in the catch of sciaenids, cephalopods and ribbon fishes in the two nets.

Studies on radiation pasteurisation of medium fatty fish 1. Control of radiation induced oxidative changes in white pomfret (Stromateus cinereus) by vacuum packaging

Marked differences were observed in proximate biochemical compositions of the skin and muscle of white pomfret. The skin showed comparatively higher content of extractable lipids and was more susceptible to radiation-induced oxidative changes like development of rancid odours and yellow discolouration than the muscle. Irradiation of skin samples under vacuum suppressed these changes. The present paper also reports on the efficacy of vacuum packaging in controlling oxidative rancidity and yellow discolouration in white pomfret skin subjected to irradiation and subsequent storage at 0-2°C.