Confocal laser scanning microscopy elucidation of the micromorphology of the leaf cuticle and analysis of its chemical composition

Electron microscopy techniques such as transmission electron microscopy (TEM) and scanning electron microscopy (SEM) have been invaluable tools for the study of the micromorphology of plant cuticles. However, for electron microscopy, the preparation techniques required may invariably introduce artefacts in cuticle preservation. Further, there are a limited number of methods available for quantifying the image data obtained through electron microscopy. Therefore, in this study, optical microscopy techniques were coupled with staining procedures and, along with SEM were used to qualitatively and quantitatively assess the ultrastructure of plant leaf cuticles. Leaf cryosections of Triticum aestivum (wheat), Zea mays (maize), and Lupinus angustifolius (lupin) were stained with either fat-soluble azo stain Sudan IV or fluorescent, diarylmethane Auramine O and were observed under confocal laser scanning microscope (CLSM). For all the plant species tested, the cuticle on the leaf surfaces could be clearly resolved in many cases into cuticular proper (CP), external cuticular layer (ECL), and internal cuticular layer (ICL). Novel image data analysis procedures for quantifying the epicuticular wax micromorphology were developed, and epicuticular waxes of L. angustifolius were described here for the first time. Together, application of a multifaceted approach involving the use of a range of techniques to study the plant cuticle has led to a better understanding of cuticular structure and provides new insights into leaf surface architecture.

Discrimination of origin of farmed trout by means of biometrical parameters, fillet chemical composition and flavour volatile compounds

To date it is well known that the quality of farmed trout is affected by diet composition, by feeding regime, by husbandry practices and by rearing conditions and environment. The trout processing industry and the large-scale retail trade, in consideration of the wide variability of trout quality and characteristics, have imposed, or will soon impose, quality criteria for the end product. Moreover, recent food scares and the malpractices of some food producers have increased public requests
for traceability. The aim of the present study was to evaluate the main chemical quality and the biometrical characteristics of rainbow trout produced in three different farms in Italy (two intensive farms, located one on mountain and one on plain, and an extensive farm in which fish fed only on naturally available nutrients) and to establish whether farmed trout
origins could be differentiated by these parameters. Trout farmed in the intensive mountain farm (IMF) showed the highest crude lipid content in the fillets and the fatty acids of their fillets were characterized by the highest percentage of MUFA. Trout farmed in the intensive plain farm (IPF) were characterized by low dressing percentage, and the lipid of their fillets
was rich in n-6 fatty acids. Trout stocked for the last year of their life in the extensive farm (EF) were leaner both in the carcass and in the fillets. The analysis of flavor volatile compounds showed some differences in the bouquet design, particularly differences in the amounts of n-3 and n-6 derivates volatile aldehydes and alcohols. All data significantly different
(P<0.05) were subjected to Linear Discriminant Analysis (LDA) and 8 variables were chosen to create two discriminant equations generating a strong prediction model for classification of farmed trout respective to their origins.

Effects of alternative dietary lipid sources on performance, tissues chemical composition, mitochondrial fatty acid oxidation capabilities, and sensory characteristics in brown trout (Salmo trutta L.)

The efficiency of five dietary lipid sources (fish oil as control—C; canola oil—CO; poultry fat—PF; pork lard—PL; and oleine oil—OO) were evaluated in juvenile brown trout (58.4±0.7 g) in an experiment conducted over 70 days at 14.6±0.4 °C. The best growth was observed in fish fed the C diet whereas the PL diet fed fish had the best feed utilization. Significant differences in carcass and muscle proximate composition, but not in liver, were noted among fish fed the different dietary treatments. The fatty acid composition of muscle largely reflected that of the diets, while total cholesterol was not affected. The atherogenicity and the thrombogenicity qualities of the trout flesh were modified by the lipid sources. Sensory analysis did not show any significant differences among the cooked fillets with respect to dietary treatments, while in uncooked products, some significant differences were observed. The carnitine palmitoyltransferase I and II (CPT-I and CPT-II) activities of liver and white muscle were assayed for a better understanding of the potential β-oxidation capability of the different dietary lipid sources. The hepatic, but not white muscle CPT-I and CPT-II activities were affected by dietary treatments. This study showed that alternative lipid sources could be used effectively for oil coating extruded diets for brown trout.

Effects of dietary lipid source on fillet chemical composition, flavour volatile compounds and sensory characteristics in the freshwater fish tench (Tinca tinca L.)

In this study the effects of soybean and linseed oils on chemical and sensory characteristics of fillets were evaluated in the freshwater fish tench (Tinca tinca L.). Five experimental diets, differing only in the relative amount of soybean and linseed oil, were formulated and the experiment was conducted on 360 sub-adult tench for 12 weeks. The fatty acid composition of muscle reflected that of the diets and significant correlations were observed. Diets containing higher amounts of n − 6 fatty acids were responsible for an increased level of n − 6 fatty acids in the fish flesh. Consequently, an increase in the relative amount of n − 6-derived volatile aldehydes was also observed. These latter compounds are generally reported to contribute negatively to the general aroma of fish muscle and, consistently, the results of the sensory analysis showed a high value for the “off-flavour” attribute for fish fed the diet containing only n − 6-rich soybean oil.

Chemical composition and tissue energy density of the cuttlefish (Sepia apama) and its assimilation efficiency by Diomedea albatrosses.

The cuttlefish Sepia apama Gray (Mollusca: Cephalopoda) is a seasonally abundant food resource exploited annually by moulting albatrosses throughout winter and early spring in the coastal waters of New South Wales, Australia. To assess its nutritional value as albatross forage, we analysed S. apama for water, lipid protein, ash contents, energy density and amino acid composition. Because albatrosses consistently consume S. apama parts preferentially in the order of head, viscera and mantle, we analysed these sections separately, but did not identify any nutritional basis for this selective feeding behaviour. The gross energy value of S. apama bodies was 20.9 kJ/g dry mass, but their high water content (>83%; cf <70% for fish) results in a relatively low energy density of 3.53 kJ/g. This may contribute to a need to take large meals, which subsequently degrade flight performance. Protein content was typically >75% dry mass, whereas fat content was only about 1%. Albatrosses feed on many species of cephalopods and teleost fish, and we found the amino acid composition of S. apama to be comparable to a range of species within these taxa. We used S. apama exclusively in feeding trials to estimate the energy assimilation efficiency for Diomedea albatrosses. We estimated their nitrogen-corrected apparent energy assimilation efficiency for consuming this prey to be 81.82 ± 0.72% and nitrogen retention as 2.90 ± 0.11 g N kg^-1 d^-1. Although S. apama has a high water content and relatively low energy density, its protein composition is otherwise comparable to other albatross prey species. Consequently, the large size and seasonal abundance of this prey should ensure that albatrosses remain replete and adequately nourished on this forage while undergoing moult.