Photometric properties of Mars soils analogs

We have measured the bidirectional reflectance of analogs of dry, wet, and frozen Martian soils over a wide range of phase angles in the visible spectral range. All samples were produced from two geologic samples: the standard JSC Mars-1 soil simulant and Hawaiian basaltic sand. In a first step, experiments were conducted with the dry samples to investigate the effects of surface texture. Comparisons with results independently obtained by different teams with similar samples showed a satisfying reproducibility of the photometric measurements as well as a noticeable influence of surface textures resulting from different sample preparation procedures. In a second step, water was introduced to produce wet and frozen samples and their photometry investigated. Optical microscope images of the samples provided information about their microtexture. Liquid water, even in relatively low amount, resulted in the disappearance of the backscattering peak and the appearance of a forward-scattering peak whose intensity increases with the amount of water. Specular reflections only appeared when water was present in an amount large enough to allow water to form a film at the surface of the sample. Icy samples showed a wide variability of photometric properties depending on the physical properties of the water ice. We discuss the implications of these measurements in terms of the expected photometric behavior of the Martian surface, from equatorial to circum-polar regions. In particular, we propose some simple photometric criteria to improve the identification of wet and/or icy soils from multiple observations under different geometries.

Visualization of cell microtubules in their native state.

BACKGROUND INFORMATION Over the past decades, cryo-electron microscopy of vitrified specimens has yielded a detailed understanding of the tubulin and microtubule structures of samples reassembled in vitro from purified components. However, our knowledge of microtubule structure in vivo remains limited by the chemical treatments commonly used to observe cellular architecture using electron microscopy. RESULTS We used cryo-electron microscopy and cryo-electron tomography of vitreous sections to investigate the ultrastructure of microtubules in their cellular context. Vitreous sections were obtained from organotypic slices of rat hippocampus and from Chinese-hamster ovary cells in culture. Microtubules revealed their protofilament ultrastructure, polarity and, in the most favourable cases, molecular details comparable with those visualized in three-dimensional reconstructions of microtubules reassembled in vitro from purified tubulin. The resolution of the tomograms was estimated to be approx. 4 nm, which enabled the detection of luminal particles of approx. 6 nm in diameter inside microtubules. CONCLUSIONS The present study provides a first step towards a description of microtubules, in addition to other macromolecular assemblies, in an unperturbed cellular context at the molecular level. As the resolution appears to be similar to that obtainable with plunge-frozen samples, it should allow for the in vivo identification of larger macromolecular assemblies in vitreous sections of whole cells and tissues.

Parallel Peptidome and Transcriptome Analyses of Amphibian Skin Secretions Using Archived Frozen Acid-Solvated Samples

Amphibian skin secretions are unique sources of bioactive peptides and their donor species are currently rapidly disappearing from the biosphere. Here, we report that both peptides and polyadenylated mRNAs from skin granular glands remain amenable to study in samples of stimulated skin secretions following their storage in 0.1 % aqueous trifluoroacetic acid at -20 °C for many years. Frozen acidified solutions of toad (Bombina variegata) skin secretions, stored for 12 years, were thawed and samples removed for direct reverse phase HPLC fractionation. Additional samples were removed, snap frozen and lyophilised for construction of cDNA libraries following polyadenylated mRNA capture using magnetic oligo-dT beads and reverse transcription. Using the bombesin and bradykinin peptides found in bombinid toad skin as models, individual variant peptides of each type were located in reverse phase HPLC fractions and their corresponding biosynthetic precursor-encoding mRNA transcripts were cloned from the cDNA library using a RACE PCR strategy. This study illustrates unequivocally that both amphibian skin secretion peptides and their biosynthetic precursor-encoding polyadenylated mRNAs are stable in frozen acid-solvated skin secretion samples for considerable periods of time-a finding that may have fundamental implications in the study of archived materials but also in the wider field of molecular biology.

Astaxanthin profiles and corresponding colour properties in Australian farmed black tiger prawn (Penaeus monodon) during frozen storage

The colour of commercial cooked black tiger prawns (Penaeus monodon) is a key quality requirement to ensure product is not rejected in wholesale markets. The colour, due to the carotenoid astaxanthin, can be impacted by frozen storage, but changes in colour or astaxanthin profile, during frozen storage, have not been studied in detail. Subsequently in this study, the aims were to define the astaxanthin (as cis, trans, mono-ester and di-ester forms) content, together with the colour properties, in both pleopods (legs) and abdominal segments. Changes in astaxanthin content and colour properties were further determined during frozen storage (−20°C). Total astaxanthin content was seen to decrease in all samples over time, with the rate of degradation being significantly greater (P < 0.05) in pleopods than abdomen. In both pleopods and abdomen, rate of degradation of esterified forms was significantly greater (P < 0.05) than non-esterified forms. Hue angle (increase), a* value (decrease) and L value (increase) were all seen to significantly change (P < 0.05) during storage, with changes being more prevalent in the pleopods. The pleopods are the key indicator of astaxanthin and colour loss in cooked black tiger prawns and preservation strategies are required to preserve astaxanthin and colour during frozen storage.

Influence of chilled and frozen storage on the stability of trout and herring fillet

Effects of chilled and frozen storage on specific enthalpy (ΔH) and transition temperature (Td) of protein denaturation as well as on selected functional properties of muscle tissue of rainbow trout and herring were investigated. The Td of myosin shifted from 39 to 33 °C during chilling of trout post mortem, but was also influenced by pH. Toughening during frozen storage of trout fillet was characterized by an increased storage modulus of a gel made from the raw fillet. Differences between long term and short term frozen stored, cooked trout fillet were identified by a compression test and a consumer panel. These changes did not affect the Td and ΔH of heat denaturation during one year of frozen storage at –20 °C. In contrast the Td of two myosin peaks of herring shifted during frozen storage at –20 °C to a significant lower value and overlaid finally. Myosin was aggregated by hydrophobic protein-protein interactions. Both thermal properties of myosin and chemical composition were sample specific for wild herring, but were relative constant for farmed trout samples over one year. Determination of Td was very precise (standard deviation <2 %) at a low scanning rate (≤ 0.25 K·min-1) and is useful for monitoring the quality of chilled and frozen stored trout and herring.

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.

Sampling of frozen prawns for bacteriological estimation

Experiments were conducted to study the significance of difference between samples taken from the surface and interior of a frozen shrimps block, as well as to determine the size of sample necessary to represent the whole block, with respect to bacterial count determination. The results showed that the surface samples and interior samples did not differ significantly at 5% level of significance and that the minimum quantity representative of the block was 21-26 gms in the case of a block weighing about 1300 gms. The procedure adopted for taking the bacterial count was the normal standard plate count method.

Postmortem changes in hilsa fish. Pt. 2. Studies on physical and bacteriological changes in iced and frozen stored hilsa fish (Tenualosa ilisha Ham.)

Studies were conducted to evaluate the quality of hilsa fish during icing and freezing storage at -20°C by determining organoleptic and bacteriological aspects. The fishes stored in ice were organoleptically in acceptable condition 2 for 20 days. The bacterial load in muscles of 4 days ice stored fish was 2.5x10² CFU/g which gradually increased up to 1.8x10⁵ CFU/g after 20 days when the fishes were organoleptically in acceptable condition. The keeping qualities of different days of ice stored fishes were also evaluated during their subsequent frozen storage at -20°C. Both 4 and 7 days of ice stored fishes were organoleptically in acceptable condition up to 48 weeks but the highest degree of freshness was found for fish stored in ice for 4 days before freezing at -20°C. The result indicates that the longer is the duration of ice storage before freezing, the shorter is the shelf life of the fish. The initial bacterial load prior to freezing of the 4 and 7 days of ice stored samples were 2.5x10³ CFU/g and 3.8x10⁴ CFU/g, respectively which reduced to 2.21x10² CFU/g and 2.38x10² CFU/g, respectively at the end of the 24 weeks of frozen storage. However, after 40 weeks the bacterial load in the frozen stored sample fell below the detection level.

Phosphate treatment of frozen prawns. Pt 2. Frozen storage characteristics of prawn meat treated with polyphosphates

This communication reports the changes in physical, organoleptic and biochemical characteristics of prawn meat dip-treated with alkaline and neutral solutions of polyphosphates during frozen storage. Results are presented on changes in thawed and cooked yields, water extractable nitrogen, non-protein nitrogen, free amino-nitrogen, salt solubility, myosin and moisture in the muscle and loss of soluble nitrogenous constituents in thaw drip during frozen storage up to seven months. The salt solubility remained unchanged during storage in samples treated with neutral polyphosphate solutions and the organoleptic quality was superior to control sample. It is concluded that dip treatment with neutralized solutions of tripolyphosphate not only maintains correct drained weight and improves cooked yield during prolonged frozen storage but also protects the frozen product from denaturation as measured by the salt solubility of the proteins.

Iced and frozen storage characteristics of cultured Chanos chanos (Forskal)

Freshly harvested milk fish (Chanos chanos) were stored in crushed ice and their storage life estimated by following biochemical, bacteriological and organoleptic changes occurring during storage. Samples of the fish were withdrawn at various intervals of storage, quick frozen, glazed and held in frozen storage at-l8°C. Shelf-life in frozen storage was determined in relation to period of ice storage prior to freezing by determining biochemical and organoleptic characteristics up to 30 weeks.

Denaturation of Labeo rohita (Rohu) actomyosin on frozen storage: preventive effect of carbohydrates

The preventive effect of sucrose and glucose on the denaturation of frozen rohu actomyosin at -20°C for 7 weeks was examined using an in vitro test model. The rate of denaturation was followed by estimating percentage salt extractability, Ca¹²+ ATPase activity and the clearing response test. Sucrose and glucose showed cryoprotective action for all concentration of actomyosin. Higher actomyosin concentration was preserved better than lower concentration. Post-rigor actomyosin was preserved to a greater extent than pre-rigor actomyosin. Correlation between percentage salt extractability and enzyme activity could not be observed in all samples of frozen actomyosin studied.

Studies on frozen storage characteristics of sole fish Cynoglossus macrolepidotus

Fresh sole fish (Cynoglossus macrolepidotus) was quick frozen at -40°C and stored at -18°C. Shelf life was evaluated by following biochemical, bacteriological and organoleptic changes occurring during storage. Rapid decrease was noted in the water extractable nitrogen and salt soluble nitrogen fractions. Samples of frozen sole fish remained in acceptable condition for 20 weeks.

Changes in the muscle of three Indian major carps during frozen storage

Chunks of Labeo rohita, Cirrhinus mrigala and Catla catla wrapped in polythene film were stored at -8 to -10°C in the freezer cabinet of the refrigerator. It was found that L. rohita and C. mrigala were acceptable up to 33 days and C. catch up to 35 days. Total volatile base nitrogen, free fatty acids and degree of sponginess of the samples showed increasing trend during frozen storage.

The effect of washing on the quality of minced catfish during frozen storage

The effect of washing minced catfish in water, sodium chloride solution (1%) and ascorbic acid solution (0.1%) in improving the quality and frozen shelf-life has been studied. Washing improved the colour and reduced the non-protein nitrogen contents and extractable nitrogen. Denaturation was more in samples washed in salt and ascorbic acid solutions. Rancidity as measured by PV and organoleptic studies showed significant reduction in washed samples. The frozen storage life was significantly enhanced by washing.

Occurrence of enterotoxigenic staphylococci in frozen fishery products

Sixty six samples of frozen fishery products consisting of frozen crab meat, frozen prawns, frozen cuttlefish and frozen mussel collected from nine cold stores situated in and around Cochin city were examined for coagulase positive staphylococci. Forty eight samples (72.72%) revealed the presence of coagulase positive staphylococci, which were then tested for enterotoxigenity, employing cellophane-over-agar method for toxin production and optimum-sensitivity-plate (OSP) method for toxin detection. Seventy seven out of one hundred and two (75.49%) coagulase positive staphylococcal isolates from the above products produced enterotoxins A, B, C, D and E either singly or in combinations. Enterotoxin A and D were detected more often than others. The occurrence of enterotoxigenic staphylococci in frozen fishery product is reported for the first time in this country.