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.

Saline water intrusion from deep artesian sources in McGregor Isles area of Lee County, Florida

Upward leakage of saline water from an artesian aquifer below 1,500 feet has caused an increase in chloride concentration in the lower Hawthorn aquifer from less than 1,000 mg/1 (milligrams per liter) to values ranging from about 1,300 to 15,000 mg/1. Similarly the higher temperatures of the intruding water has caused an increase in water temperatures in the aquifer from 82"F to values ranging from 83 to 93"F. The intruding water moves upward either through the open bore hole of deep wells or test holes, or along a fault or fracture system, which has been identified in the area. From these points of entry into the lower Hawthorn aquifer, the saline water spreads laterally toward the south and southeast, but is generally confined to components of the fault system. The saline water moves upward from the lower Hawthorn aquifer into the upper Hawthorn aquifer through the open bore hole of wells, which connect the aquifers. This movement has resulted in an increase in chloride from less than 200 mg/1 in the unaffected parts of the upper Hawthorn aquifer to values commonly ranging from about 300 to more than 3,000 mg/1 in parts of the aquifer affected by upward leakage. The upper Hawthorn aquifer is the principal source of ground-water supply for public water-supply systems in western Lee County. Similar effects have been noted in the water-table aquifer, where chloride increased from less than 100 to concentrations ranging from about 500 to more than 5,000 mg/1. This was caused by the downward infiltration of water discharged at land surface from wells tapping the lower Hawthorn aquifer. The spread of saline water throughout most of the McGregor Isles area is continuing as of 1971. (40 page document)

Optimizing the benefits from Nigerian small and medium size reservoirs through cooperative management approach

The paper traces the different management practices adopted for Nigerian inland water bodies from the Colonial era to independence. It observes that the full potentials of these waters have never been realized over the years due to the absence of an effective management. The replacement of the traditional fisheries management by the centralized top-down approach by government after independence has not helped matters. Lately, the cooperative/community-based management approach has taken the centre stage worldwide. This has been identified to offer the most viable and equitable option towards the attainment of an optimum utilization of the fisheries resource. The entire community sensing security of tenure and enjoying some of the benefits from access control will actively take responsibility and enforcement. The paper drew experiences from some water bodies in Bangladesh, Philippines, Benin Republic and Malawi showing sound management strategy that, if adopted for our small and medium size reservoirs and other water bodies, would help optimize on an sustainable manner the benefits from those water bodies

Subspecies, morphs and clines in the amphipod Gammarus duebeni from fresh and saline waters

The nature and extent of morphological variation within populations of Gammarus duebeni are examined. The exceptional tolerance of G. duebeni to salinities that encompass three orders of magnitude was known in the 19th Century, and has attracted considerable attention from physiologists and ecologists in the 20th Century, including the likelihood that populations in freshwater are distinct from those living in more saline environments. It is concluded that gradual evolution into discrete and readily distinguished subspecies is currently underway, eventually producing several new species in freshwater and saline habitats.

Mastigophora and Rhizopoda found in saline lakes Weissovo and Repnoie (near Slaviansk). Selected passages describing Ochromonas species and Pedinella. [Translation of: Trudy Obshch.Ispyt.Prir.imp.khar'kov.Univ. 21 119-140, 1888]

This translation includes selected passages of a longer paper on Mastigophora and Rhizopoda found in saline lakes Weissovo and Repnoie. The translation focuses on describing taxonomy and morphology of Ochromonas species and Pedinella. Plates and figures of the original paper are not included in the translation.

Preliminary observation on culture of Penaeus monodon in low-saline waters

The feasibility of semi-intensive culture of Penaeus monodon in low saline environment was investigated by comparing the growth and production in low (0.16-6.52 ppt) and high (4.60-19.42 ppt) saline areas at two stocking densities (10.5 and 16 individuals/m super(2)). After 135 days of culture, yield of shrimp in low and high stocking densities was 1563.37 kg/ha and 2274 kg/ha, respectively, in low saline ponds, and 1173.00 and 1974.00 kg/ha, respectively in high saline ponds. Food conversion ratio (FCR, 1.31-1.58) and specific growth rate (SGR 21.04-21.19%) were higher in low saline ponds as compared to high saline ponds (FCR, 1.35-1.68; SGR, 19.22-19.88%). Growth of shrimp was satisfactory in low saline ponds even when salinity decreased after 60 days of culture to almost freshwater level (0.16 ppt) indicating the variability of semi-intensive culture of P. monodon in low saline environment.

Tilapia broodstock development for saline waters in the Philippines

Four Oreochromis species were used in the study. Progenies from the 27 cross combinations (5 pure breds and 22 crossbreds) were evaluated in 10 environments with different salinity levels and agro-climatic conditions using communal rearing concept. Among the different cross combinations reared across environments, O. aureus x O. spilurus gave the highest body weight and O. mossambicus x O. spilurus, the highest survival rate. Positive percent mean heterosis were observed in the crosses between O. mossambicus x O. niloticus, FAC selected line and O. aureus x O. spilurus.

Management of waterlogged areas, island and coastal saline soils using fish based mixed farming system

Problems in India regarding the management of various coastal saline soil and waterlogged environments are discussed in detail, considering in particular the potential application of mixed fish farming systems. Various operational and cost requirements of such systems are examined.

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.

Physical soil characteristics of waterlogged and saline lands of five districts of Haryana, India

Physical characteristics of waterlogged and saline soils of five districts, namely Rohtak, Faridabad, Gurgaon, Sonipat and Jhajjar, of Haryana were studied to evaluate the aquaculture potential. The soil samples from 76 sites were procured and analysed for pH, electrical conductivity, soil particles and water retention capacity, since the knowledge of these parameters is essential for the preliminary evaluation of a site for aqua-farming. Six soil types were identified and clay-loam was observed to be the maximum at 45 (59.21%) sites, followed by silty-clay loam at 14 (18.42%), sandy-clay loam at six (7.89%), silty-loam at four (5.26%), sandy-loam at four (5.26%) and loam at three (3.94%) sites. The pH of soil indicated moderate to high alkaline conditions ranging between 8.01and 9.00 at 53 (69.73%) sites, 9.01 and 10.00 at 17 (22.36%) sites, and low between 7.01 and 8.00 at six (7.89%) sites. The electrical conductivity was found to range between <1 and 10 mmho cmˉ¹ with the value of <1.0 mmho cmˉ¹ at 36 (47.36%) sites. The water retention capacity was observed mostly to be moderate. The paper describes the interrelationship between these parameters with reference to the suitability for aqua-farming.

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.

Studies on radiation pasteurisation of medium fatty fish 3. Storage properties of white pomfret (Stromateus cinereus) fillets

White pomfret fillets packed under aerobic conditions had a limited shelf life of 8 days as against 10 days for samples packed under vacuum and stored at 0-2°C. Irradiation and subsequent storage of the fillets under vacuum at 0-2°C exhibited shelf lives of 30, 50 and 60 days for radiation doses of 0.1, 0.3 and 0.5 Mrad respectively in contrast to aerobically packed fillets which showed only 20, 35 and 50 days of storage life for the same levels of radiation doses and developed yellow discolouration and rancid odours.