Enzyme assisted extraction of chitin from shrimp shells (Litopenaeus vannamei)

BACKGROUND: Chemical chitin extraction generates large amounts of wastes and increases partial deacetylation of the product. Therefore, the use of biological methods for chitin extraction is an interesting alternative. The effects of process conditions on enzyme assisted extraction of chitin from the shrimp shells in a systematic way were the focal points of this study. RESULTS: Demineralisation conditions of 25C, 20 min, shells-lactic acid ratio of 1:1.1 w/w; and shells-acetic acid ratio of 1:1.2 w/w, the maximum demineralisation values were 98.64 and 97.57% for lactic and acetic acids, respectively. A total protein removal efficiency of 91.10% by protease from Streptomyces griseus with enzyme-substrate ratio 55 U/g, pH 7.0 and incubation time 3 h is obtained when the particle size range is 50-25 μm, which was identified as the most critical factor. The X-ray diffraction and 13C NMR spectroscopy analysis showed that the lower percent crystallinity and higher degree of acetylation of chitin from enzyme assisted extraction may exhibit better solubility properties and less depolymerisation in comparison with chitin from the chemical extraction. CONCLUSION: The present work investigates the effects of individual factors on process yields, and it has shown that, if the particle size is properly controlled a reaction time of 3 h is more than enough for deproteination by protease. Physicochemical analysis indicated that the enzyme assisted production of chitin seems appropriate to extract chitin, possibly retaining its native structure.

Microcapsules with improved shells

Disclosed are microcapsules and methods for preparing and using them, as well as methods for improving various properties of microcapsules like impermeability.

Fracture analysis of spherical shells : application to cracking of macadamia nuts

The main consideration in recovering the macadamia kernal is to crack the spherical nutshell without damaging the kernal. Five mechanical cracking tools were tested, and the fracture mechanisms of nutshells, under various cracking loads, were studied. A classical theoretical approach and a numerical method were both used to investigate the influence of crack face closure on the stress intensity factor for a cracked spherical shell subjected to membrane forces and bending moments.

Encapsulation of single hMSCs in polyelectrolyte shells

The main objective of this Thesis was to encapsulate single viable cells within polyelectrolyte films using the Layer-by-Layer (LbL) technique. Most of the experiments used human mesenchymal stem cells (MSCs) whose characteristics (capacity of selfrenewal and potential to differentiate into several types of cells) make them particularly interesting to be used in biomedical applications. Also, most of the experiments used alginate (ALG) as the anionic polyelectrolyte and chitosan (CHI) or poly(allylamine hydrochloride) (PAH) as the cationic polyelectrolyte. Hyaluronic acid (HA) was also tested as an anionic polyelectrolyte. At the beginning of the work, the experimental conditions necessary to obtain the encapsulation of individual cells were studied and established. Through fluorescence microscopy visualization by staining the cell nucleus and using polyelectrolytes conjugated to fluorescent dyes, it was possible to prove the obtainment of capsules containing one single cell inside. Capsules aggregation was an observed problem which, despite the efforts to design an experimental process to avoid this situation (namely, by playing with cell concentration and different means of re-suspending and stirring the cells), was not completely overcome. In a second part of the project, single cells were encapsulated within polyelectrolyte layers made of CHI/ALG, PAH/ALG and PAH/HA and their viability was evaluated through the resazurin reduction assay and the Live/Dead assay. In these experiments, during the LbL process, polyelectrolyte solutions were used at a concentration of 1mg/mL based on literature. In general, the viability of the encapsulated cells was shown to be very low/absent. Then, as a consequence of the lack of viability of cells encapsulated within polyelectrolyte layers, the LbL technique was applied in cells growing adherent to the surface of cell culture plates. The cells were cultured like in a sandwich, between the surface of the cell culture dish and the polyelectrolyte layers. Also here, the polyelectrolyte solutions were used at a concentration of 1mg/mL during the LbL process. Surprisingly, cell viability was also absent in these systems. A systematic study (dose-effect study) was performed to evaluate the effect of the concentration of the individual polyelectrolytes (ALG, CHI and PAH were studied) in cell viability. Experiments were performed using cells growing adherent to the surface of cell culture plates. The results pointed out that a very high (cytotoxic) concentration of polyelectrolytes had been in use. Also, in general, PAH was much more cytotoxic than CHI, whereas ALG was the less cytotoxic polyelectrolyte. Finally, using alginate and chitosan solutions with adequate concentrations (low concentrations: 50ng/mL and 1μg/mL), the encapsulation of single viable cells was again attempted. Once again, the encapsulated cells were not shown to be viable. In conclusion, the viability of the encapsulated cells is not only dependent on the cytotoxic characteristics (or combined cytotoxic characteristics) of the polyelectrolytes but it seems that, when detached from the culture plates, the cells become too fragile and lose their viability very easily.

Are the preference and selection patterns of hermit crabs for gastropod shells species- or site-specific?

Experimental analyses of hermit crabs and their preferences for shells are essential to understand the intrinsic relationship of the crabs' dependence on shells, and may be useful to explain their shell use pattern in nature. The aim of this study was to evaluate the effect of crab species and site on the pattern of shell use, selection, and preference in the south-western Atlantic hermit crabs Pagurus brevidactylus and Pagurus criniticornis, comparing sympatric and allopatric populations. Differently from the traditional approach to evaluate shell preference by simply determining the shell selection pattern (i.e., the number of shells of each type selected), preference was defined (according to [Liszka, D., Underwood, AJ., 1990. An experimental design to determine preferences for gastropod shells by a hermit-crab. J. Exp. Mar. Biol. Ecol., 137(1), 47-62]) by the comparison of the number of crabs changing for a particular shell type when three options were given (Cerithium atratum, Morula nodulosa, and Tegula viridula) with the number of crabs changing for this same type when only this type was offered. The effect of crab species was tested at Cabelo Gordo Beach, where P. brevidacrylus was found occupying shells of C. atratum, M. nodulosa, and T viridula in similar frequencies, whereas P. criniticornis occupied predominantly shells of C atratum. In laboratory experiments the selection patterns of the two hermit-crab species for these three gastropods were different, with P criniticornis selecting mainly shells of C atratum, and R brevidactylus selecting more shells of M. nodulosa. The shell preference was also dependent on crab species, with P. criniticornis showing a clear preference for shells of C atratum, whereas P. brevidactylus did not show a preference for any of the tested shells. The effect of site was tested for the two species comparing data from Cabelo Gordo to Preta (P brevidactylus) and Araca beaches (P. criniticornis). The pattern of shell use, selection, and preference was demonstrated to be dependent on site only for P. brevidactylus. The results also showed that the shell use pattern of P criniticornis can be explained by its preference at both sites, whereas for P. brevidactylus it occurred only at Cabelo Gordo, where the absence of preference was correlated with the similar use of the three gastropod species studied. Finally, the results showed that the shell selection pattern cannot be considered as a measure of shell preference, since it overestimates crab selectivity. (C) 2009 Elsevier B.V. All rights reserved.

Taphonomy of Bouchardia rosea (Rhynchonelliformea, Brachiopoda) shells from Ubatuba Bay, Brazil: implications for the use of taphonomic signatures in (paleo)environmental analysis

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Penetration time of Salmonella Heidelberg through shells of white and brown commercial eggs

This study aimed at determining the minimum time required for the penetration of Salmonella Heidelberg inside the eggs after contact with contaminated material. Recently-collected brown and white eggs from laying hens between 45-50 weeks of age, reared in a commercial poultry house, were artificially contaminated by contact with wood shavings moistened with liquid inoculum of Salmonella Heidelberg in stationary-growth phase (10³-10(4) CFU g-1). According to type (white or brown), eggs were distributed into three different groups, with four replicates each: negative control group (no artificial contamination), positive control group (analyzed externally immediately after contamination and internally after the maximum storage period of the test group) and test group. Eggs were stored at controlled environmental temperature varying from 25ºC to 30ºC. In the test group, eggs contents (yolk and albumen) were pooled and analyzed after 1:00, 1:30, 2:00, 2:30, 3:00, 3:30, and 4:00 hours after contamination for the presence of Salmonella Heidelberg in 25g of this pool. The experimental unit consisted of five eggs in each test. The analysis protocol included pre-enrichment, selective enrichment, plating on selective agar, and biochemical and serological tests. The results obtained were submitted to logistic regression, which indicated that the presence of Salmonella Heidelberg was verified after 2:16 h and 2:44 h of contact with white and brown eggs, respectively.

Biotic interactions recorded in shells of recent rhynchonelliform brachiopods from San Juan Island, USA

Biotic interactions between brachiopods and spionid polychaete worms, collected around San Juan Islands (USA), were documented using observations from live-collected individuals and traces of bioerosion found in dead brachiopod shells. Specimens of Terebratalia tranversa (Sowerby), Terebratulina unguicula (Carpenter), Laqueus californianus (Koch), and Hemithiris psittacea (Gmelin) were collected from rocky and muddy substrates, from sites ranging from 14.7-93.3 m in depth. Out of 1,131 specimens, 91 shells showed traces of bioerosion represented by horizontal tubes. Tubes are U-shaped, straight or slightly curved, sometimes branched, with both tube openings communicating externally. on internal surfaces of infested shells, blisters are observed. All brachiopod species yielded tubes, except for H. psittacea. Tubes are significantly more frequent on live specimens, and occur preferentially on larger, ventral valves. This pattern suggests selectivity by the infester rather than a taphonomic bias. Given the mode of life of studied brachiopods (epifaunal, sessile, attached to the substrate, lying on dorsal valve), ventral valves of living specimens should offer the most advantageous location for suspension-feeding infesters. Frequent infestation of brachiopods by parasitic spionids is ecologically and commercially noteworthy because farmed molluscs are also commonly infested by parasitic polychaetes. In addition, brachiopod shells are among the most common marine macroscopic fossils found in the Phanerozoic fossil record. From a paleontological perspective, spionid-infested brachiopod shells may be a prime target for studying parasite-host interactions over evolutionary time scales.

Drill holes in shells of Permian benthic invertebrates

Newly discovered benthic fossils and specimens illustrated in the paleontological literature indicate that drilling predators (or parasites) were present in the Permian. New field data from southern Brazil document the first drill holes ever reported for Permian bivalve mollusks. In addition, a literature review revealed drill holes in shells of articulate brachiopods from Russia, Greece, and West Texas. Holes range in size from 0.1 to 5.8 mm and are typically round, cylindrical, singular penetrations perpendicular to the valve surface. Incomplete, healed, and multiple holes are absent. Drilling frequency, a proxy for predation intensity, is very low: less than 1 percent (this estimate may be seriously affected by taphonomic and monographic biases). Literature data suggest that frequency of drilled specimens varied significantly among higher brachiopod taxa. The geography and stratigraphy of drilled specimens indicate that drilling organisms were worldwide in their occurrence and continuously present in marine ecosystems throughout the Permian. This report is consistent with other recent studies indicating that although drillers were continuously present throughout the Phanerozoic, drilling intensity was lower in the Late Paleozoic and early Mesozoic.

Utilization of shells of the snail Achatina fúlica Bowdich, 1822 (Mollusca, Gastropoda) by the hermit crab Clibanarius vittatus (Bosc, 1802) (Decapoda, Anomura) in the São Vicente Estuary, São Paulo, Brazil

Hermit crabs depend on mollusc shells for housing. In this study, an unusual resource is reported for a hermit crab that usually inhabits marine gastropod shells. During a field study conducted from May 2001 to April 2003 in an estuarine area in São Vicente, state of São Paulo, Brazil, 21 individuals of Clibanarius vittatus (Bosc, 1802) were found inhabiting the shells of the terrestrial gastropod Achatina fullea Bowdich, 1822. A. fúlica occurs in nearshore grass patches, where occasional contact with sea water kills them, and their shells then become available to the hermit crabs.

An exact series solution for the vibration analysis of cylindrical shells with arbitrary boundary conditions

In this paper, an exact series solution for the vibration analysis of circular cylindrical shells with arbitrary boundary conditions is obtained, using the elastic equations based on Flügge's theory. Each of the three displacements is represented by a Fourier series and auxiliary functions and sought in a strong form by letting the solution exactly satisfy both the governing differential equations and the boundary conditions on a point-wise basis. Since the series solution has to be truncated for numerical implementation, the term exactly satisfying should be understood as a satisfaction with arbitrary precision. One of the important advantages of this approach is that it can be universally applied to shells with a variety of different boundary conditions, without the need of making any corresponding modifications to the solution algorithms and implementation procedures as typically required in other techniques. Furthermore, the current method can be easily used to deal with more complicated boundary conditions such as point supports, partial supports, and non-uniform elastic restraints. Numerical examples are presented regarding the modal parameters of shells with various boundary conditions. The capacity and reliability of this solution method are demonstrated through these examples. © 2012 Elsevier Ltd. All rights reserved.

Awaking the vacuum with spheroidal shells

It has been shown that well-behaved spacetimes may induce the vacuum fluctuations of some nonminimally coupled free scalar fields to go through a phase of exponential growth. Here, we discuss this mechanism in the context of spheroidal thin shells emphasizing the consequences of deviations from spherical symmetry. © 2013 American Physical Society.