Development of an in vitro model to assess protein bioavailability in diets for common octopus (Octopus vulgaris)

An in vitro model designed to assess protein bioavailability in diets for growing Octopus was developed. The model required a previous assessment of some functional features of protein digestion in this species like the main producing organs, optimum pH for activity and total production per g tissue. The main producing organs identified were the salivary glands and the hepatopancreas (HP), being optimum pH for protease activity quite different in both organs (mostly alkaline in the posterior salivary glands and acid in the HP). In spite of the high‐specific protease activity measured in the salivary glands, a major role of the HP in protein hydrolysis is suggested due to the much bigger size of this viscera. All this information was used as a basis to develop an in vitro two‐step hydrolysis process, which simulated protein hydrolysis performed by these two organs using the Octopus enzymes. The assay was used to evaluate differences in amino acid bioavailability from several protein sources (casein, gelatin, fish meal, squid meal and krill meal) that could be used as feed ingredients for this species. As significant differences were detected both in total amount and in rate of release of the amino acids from such proteins, the model is proposed as a complementary tool in the selection and nutritional evaluation of protein ingredients to be used in diets for Octopus.     

Carotenoid profile modification during refrigerated storage in untreated and pasteurized orange juice and orange juice treated with high-intensity pulsed electric fields

A comparative study was made of the evolution and modification of various carotenoids and vitamin A in untreated orange juice, pasteurized orange juice (90 degrees C, 20 s), and orange juice processed with high-intensity pulsed electric fields (HIPEF) (30 kV/cm, 100 micros), during 7 weeks of storage at 2 and 10 degrees C. The concentration of total carotenoids in the untreated juice decreased by 12.6% when the juice was pasteurized, whereas the decrease was only 6.7% when the juice was treated with HIPEF. Vitamin A was greatest in the untreated orange juice, followed by orange juice treated with HIPEF (decrease of 7.52%) and, last, pasteurized orange juice (decrease of 15.62%). The decrease in the concentrations of total carotenoids and vitamin A during storage in refrigeration was greater in the untreated orange juice and the pasteurized juice than in the juice treated with HIPEF. During storage at 10 degrees C, auroxanthin formed in the untreated juice and in the juice treated with HIPEF. This carotenoid is a degradation product of violaxanthin. The concentration of antheraxanthin decreased during storage, and it was converted into mutatoxanthin, except in the untreated and pasteurized orange juices stored at 2 degrees C.     

Drought pretreatment increases the salinity resistance of tomato plants

Agricultural productivity is worldwide subjected to increasing salinity problems. Various strategies are applied to overcome the deleterious effects of salinity on plants. This study was conducted in order to determine whether drought pretreatment of seedlings or seed pretreatment with NaCl increases the long‐term salinity resistance of tomato (Solanum lycopersicum L.) and whether the adaptive response to salinity is accompanied by physiological changes throughout the plant‐growth cycle. When plants were pretreated at the five‐leaf growth stage, the plant dry weight was significantly higher in drought‐pretreated than in non‐pretreated plants after 50 d of salt treatment. The positive effect of drought pretreatment applied at the five‐leaf stage was maintained throughout the entire growth cycle, as fruit yield of drought‐pretreated plants was 40% higher than that of non‐pretreated plants at the end of the harvest period (150 d of 70 mM NaCl treatment). Moreover, the most productive plants maintained lower Na⁺ and Cl^– accumulation in their leaves until the end of the growth cycle, which shows that adaptation is a long‐term response during which the plants adjust their physiology to the environmental conditions. Salt resistance was also improved through seed pretreatment with NaCl. In conclusion, drought pretreatment applied at the five‐leaf stage or seed pretreatment with NaCl provide an alternative way to enhance salt resistance in tomato, and the increase in yield is associated with physiological changes throughout the plant‐growth cycle.     

Development of a method for the genetic identification of mussel species belonging to Mytilus, Perna, Aulacomya, and other genera

Legislation regarding the labeling of processed products is an important issue in the protection of consumer rights. This labeling is especially important in products that cannot be identified on the basis of their morphological characters, because these are removed from the animal in the transformation process. The goal of this study was the identification of mussel species using Polymerase Chain Reaction-Restriction Fragment Length Polymorphism (PCR-RFLP) and Forensically Informative Nucleotide Sequencing (FINS) methodologies. The molecular marker selected was 18S rDNA (nuclear small-subunit rDNA gene), which allows identification at the genus level and at the species level in some cases. The genera included in this study were Mytilus, Perna, Aulacomya, Semimytilus, Brachidontes, Choromytilus, and Perumytilus. Different markers were used for genetic identification at the species level. To identify the species included in the genus Perna and Choromytilus, a fragment of ITS 1 (Internal Transcribed Spacer 1) was amplified by multiplex PCR and digested with restrictases. The species of Mytilus were identified by length polymorphism and RFLP of the polyphenolic adhesive protein gene. This methodology was validated with products manufactured in the authors' pilot plant and applied to commercial samples. Therefore, this sequential method can be completely or partially used to determine the mussel genus or species present in any food product.     

THE IMPACT OF ORGANIC AMENDMENTS ON FOREST SOIL PROPERTIES UNDER MEDITERRANEAN CLIMATIC CONDITIONS

Restoring the native vegetation is one of the most effective ways to control soil degradation in Mediterranean areas, especially in very degraded areas. In the initial months after afforestation, vegetation cover establishment and soil quality could be better sustained if the soil was amended with an external extra source of organic matter. The goal of this study was to test the effect of various organic amendments on select soil properties [soil organic carbon (SOC), pH, and electrical conductivity (EC)] over a 24‐month period. Four amendments were applied in an experimental set of plots: straw mulching (SM), mulch with chipped branches of Aleppo Pine (Pinus halepensis L.; PM), sheep manure compost (SH), and sewage sludge (RU). Plots were afforested following the same spatial pattern, and amendments were mixed with the soil at the rate 10 Mg ha⁻¹. Organic amendments helped maintain SOC over the initial 6 months after the afforestation. However, only the SM and PM treatments had increased SOC values after 24 months. Decreases in EC were found after the addition of SM, PM, and SH amendments. However, RU increased EC values 24 months after the afforestation. Variations in pH values were not sufficient to establish differences among the various treatments. Furthermore, the results show that forest soils with or without organic amendments responded similarly to the seasonal changes in Mediterranean conditions. Copyright © 2014 John Wiley & Sons, Ltd.     

Intravaginal progesterone devices in synchronization protocols for artificial insemination in beef heifers

Two experiments were designed to investigate the administration of intravaginal progesterone in protocols for oestrus and ovulation synchronization in beef heifers. In Experiment 1, cyclic Black Angus heifers (n = 20) received an Ovsynch protocol and were randomly assigned to receive (CIDR‐Ovsynch) or not (Ovsynch) a progesterone device between Days 0 and 7. Treatment with a controlled internal drug release (CIDR) device significantly increased the size of the dominant follicle prior to ovulation (12.8 ± 0.4 CIDR‐Ovsynch vs 11.4 ± 0.4 Ovsynch) (p < 0.02). Plasma progesterone concentrations throughout the experiment were affected by the interaction between group and day effects (p < 0.004). In Experiment 2, cyclic Polled Hereford heifers (n = 382) were randomly assigned to one of the six treatment groups (3 × 2 factorial design) to receive a CIDR, a used bovine intravaginal device (DIB), or a medroxiprogesterone acetate (MAP) sponge and GnRH analogues (lecirelin or buserelin). All heifers received oestradiol benzoate plus one of the devices on Day 0 and PGF on Day 7 pm (device withdrawal). Heifers were detected in oestrus 36 h after PGF and inseminated 8–12 h later, while the remainder received GnRH 48 h after PGF and were inseminated on Day 10 (60 h). The number of heifers detected in oestrus on Day 8 and conception rate to AI on Day 9 were higher (p < 0.01) in the used‐DIB than in the CIDR or MAP groups, while the opposite occurred with the pregnancy rate to FTAI on Day 10 (p < 0.01). There was no effect of progesterone source, GnRH analogue or their interaction on overall pregnancy rates (64.9%). Progesterone treatment of heifers during an Ovsynch protocol resulted in a larger pre‐ovulatory follicle in beef heifers. Progesterone content of intravaginal devices in synchronization protocols is important for the timing of AI, as the use of low‐progesterone devices can shorten the interval to oestrus.     

Effects of agricultural practices on color, carotenoids composition, and minerals contents of sweet peppers, cv. Almuden

Consumers demand organic products because they believe they are more flavorful and respectful to the environment and human health. The effects of conventional, integrated, and organic farming, grown in a controlled greenhouse, on color, minerals, and carotenoids of sweet pepper fruits ( Capsicum annuum), cv. Almuden, were studied. Experimental results proved that organic farming provided peppers with the highest (a) intensities of red and yellow colors, (b) contents of minerals, and (c) total carotenoids. Integrated fruits presented intermediate values of the quality parameters under study, and conventional fruits were those with the lowest values of minerals, carotenoids, and color intensity. As an example, the concentrations of total carotenoids were 3231, 2493, and 1829 mg kg (-1) for organic, integrated, and conventional sweet peppers, respectively. Finally, organic red peppers could be considered as those having the highest antioxidant activity of all studied peppers (agricultural farming and development stage).     

Mycobacterium bovis cultured from commercially pasteurized cows' milk: laboratory cross-contamination

The ability of Mycobacterium paratuberculosis to survive the commercial pasteurization process of raw milk remains controversial. In a study undertaken in Venezuela to culture M. paratuberculosis from commercially pasteurized cows' milk, 83-200 ml containers of milk were processed and cultured on Herrold's egg yolk slants. No M. paratuberculosis was cultured but a total of six colonies of Mycobacterium bovis were isolated from one container each from two different milk providers. Because laboratory cross-contamination was suspected, the laboratory records were reviewed and spoligotyping was carried out on the isolated individual colonies. On the day before these milk specimens were processed, the biological safety cabinet had been used for the isolation of M. bovis from lymph nodes from infected cattle. Spoligotyping showed that that the colonies isolated from the milk all had the same pattern as the strains isolated from the lymph nodes that were processed the previous day. As far as we know, this is the first report of cross-contamination in a veterinary mycobacterial laboratory. False-positive cultures in the mycobacterial laboratory are not rare. In this setting M. bovis was isolated because it is the most common manipulated organism in this laboratory. We believe that reports on the isolation of M. paratuberculosis from commercially pasteurized milk should exclude cross-contamination before reporting, especially when this organism is routinely isolated from animal material in the same lab.