Separation of olive proteins combining a simple extraction method and a selective capillary electrophoresis (CE) approach: application to raw and table olive samples

A simple extraction method was developed to extract proteins from olive samples based on chloroform/methanol extraction followed by a protein precipitation with cold acetone. Then, a capillary electrophoresis (CE) method was carried out using an acid buffer (1 M formic acid at pH 2) to ensure a positive net charge for proteins and a neutral charge for potential interferents as polyphenols. The method developed was applied to raw and table olive samples. Interestingly, raw olive samples showed differences in protein profiles depending upon the botanical variety of olives and their geographical region. Protein profiles obtained for table olives also showed differences according to the sample treatment. Thus, a signal reduction in the electropherograms obtained for black olives was observed in comparison to those achieved for treated green olives. In this work, the use of protein profiles was demonstrated to be a powerful tool for studying variations among olive samples.     

Antiadhesion and antibiofilm activities of high molecular weight coffee components against Streptococcus mutans

In previous studies we demonstrated that green and roasted coffee contains low molecular weight (LMW) compounds capable of inhibiting the ability of Streptococcus mutans, the major causative agent of human dental caries, to adhere to hydroxyapatite (HA) beads. This study addressed the ability of the whole high molecular weight coffee fraction (cHMW) and of its melanoidin and non-melanoidin components (GFC1-5), applied at concentrations that occur in coffee beverages, to (i) inhibit S. mutans growth; (ii) affect S. mutans sucrose-dependent adhesion to and detachment from saliva-coated HA beads (sHA); and (iii) inhibit biofilm development on microtiter plates. The results indicated that only cHMW is endowed with antimicrobial activity. The cHMW fraction and each of the five GFC components inhibited S. mutans adhesion, the strongest effect being exerted by cHMW (91%) and GFC1 (88%). S. mutans detachment from sHA was four times greater (～20%) with cHMW and the GFC1 and GFC4 melanoidins than with controls. Finally, biofilm production by S. mutans was completely abolished by cHMW and was reduced by 20% by the melanoidin components GFC2 and GFC4 and by the non-melanoidin component GFC5 compared with controls. Altogether these findings show that coffee beverage contains both LMW compounds and HMW melanoidin and non-melanoidin components with a strong ability to interfere in vitro with the S. mutans traits relevant for cariogenesis.     

Two-season study of the influence of regulated deficit irrigation and reflective mulch on individual and total phenolic compounds of nectarines at harvest and during storage

The influence of deficit irrigation (Deficit) and reflective mulch (Reflective) of Caldesi 2000 nectarines on the content of individual phenolic compounds was studied at harvest and during storage for 2, 4, and 6 weeks at 2 °C during two consecutive years (2007 and 2008). Individual phenolic groups in the edible fruit part consisted mainly of proanthocyanidins (200 mg/100 g fw), lower content of phenolic acids (17 mg/100 g fw), and minor content of flavonols (5 mg/100 g fw) and anthocyanins (1.2 mg/100 g fw). Deficit irrigation increased the content of total phenolics, including proanthocyanidins and phenolic acids, reaching similar amounts in both years. Sun-exposed fruit (upper part of canopy) showed higher content than shaded fruit (lower part of canopy). However, Reflective significantly increased the content of total phenolics, particularly phenolic acids and proanthocyanidins, of fruit located in the lower part of the canopy. During storage, Deficit and Reflective did not affect the content of phenolic acids, flavonols, and proanthocyanidins when compared to the content at harvest. Optimizing cultural practices can be a way to increase the phenolic content of nectarines.     

Micromonospora: An important microbe for biomedicine and potentially for biocontrol and biofuels

Micromonospora species have long been recognized as important sources of antibiotics and also for their unusual spores. However, their involvement in plant-microbe associations is poorly understood although several studies demonstrate that Micromonospora species function in biocontrol, plant growth promotion, root ecology, and in the breakdown of plant cell wall material. Our knowledge of this generally understudied group of actinomycetes has been greatly advanced by the increasing number of reports of their associations with plants, by the deployment of DNA cloning and molecular systematics techniques, and by the recent application of whole genome sequencing. Efforts to annotate the genomes of several Micromonospora species are underway. This information will greatly augment our knowledge of these versatile microorganisms.     

Stabilization of Mine Tailings Using Fly Ash and Sewage Sludge Planted with Phalaris arundinacea L.

The impact of plants (Phalaris arundinacea L.) on the leakage of ammonium, cadmium, copper, nitrate, phosphate, and zinc from sulfidic mine tailings covered with wood fly ash and sewage sludge was investigated. Either ash or sludge was placed in contact with the tailings, and ash layers of either low or high compactness were used. It was revealed that an ash/sludge cover effectively decreased the metal leaching from the tailings regardless of the order in which the materials were applied. Plants decreased the amount of leachate and the concentrations of ammonium and phosphate. The presence of ash below the sludge decreased the plant uptake of copper and zinc and nitrate leakage. However, when the ash was added as a thin (1.5 cm) porous layer, roots and air reached the tailings and caused high metal leakage. The results support the use of a vegetated ash/sludge cover in the treatment of mine tailings, provided that the sealing layer is firm enough to prevent root penetration.     

Effects of Dietary Fiber on Growth and Gastric Emptying Time of the Freshwater Prawn Macrobrachiurn rosenbergii (De Man, 1879)

Feeding experiments were conducted on large and small adult freshwater prawns Macrobrachium rosenbergii (36.96 ± 1.65 and 13.48 ± 1.65 g in weight) to determine the impact of dietary fiber on growth, digestion, and gastric emptying time. Fiber was incorporated as Q- cellulose into four semi-purified diets at calculated levels of 0, 5, 10 and 15% at the expense of starch and fed for an 8-wk period. The experiment was conducted in a closed, filtered, recirculating freshwater system. The results indicate that the specific growth rates, feed conversion ratios, and protein efficiency ratios improved as the levels of dietary fiber increased from 0.4% to 8%. Gastric emptying times increased as the level of inclusion of cellulose in the diets increased. It was concluded that diets containing up to 10% dietary fiber increases growth rates in adult prawn Macrobrachium rosenbergii by increasing nutrient residence time in the gastrointestinal tract, thereby increasing absorption.     

Type IV secretion and Brucella virulence

The type IV secretion system, encoded by the virB region, is a key virulence factor for Brucella. The 12 genes of the region form an operon that is specifically induced by phagosome acidification in cells after phagocytosis. We speculate that the system serves to secrete unknown effector molecules, which allow Brucella to pervert the host cell endosomal pathways and to create a novel intracellular compartment in which it can replicate.     

Artificial Propagation of Mutton Snapper Lutjanus analis, A New Candidate Marine Fish Species for Aquaculture

Abstract Wild-caught mutton snapper Lutjanus analis , a high-value marine food fish species, matured in flow-through seawater (36 g/L) tanks after 3 yr in captivity. On 31 May 1995, a female with a mean oocyte diameter of 382 μm was injected with human chorionic gonadotropin (HCG) (500 IU/kg body wt.) followed 24 h later by a second injection (1,000 IU/kg body wt.). At the time of the second injection, three males were injected with HCG (500 IU/ kg body wt.). Voluntary spawning occurred 33 h after the first injection, with a total of 534, 781 eggs released. Fertilization rate was 75.7%, while average diameter of fertilized eggs was 783 μm. Embryos were stocked in a 30-m³ outdoor tank at a density of 10.5/L. On day 2 post-hatching (d2ph), larval density was 8.61 larvae/L, and average notochord length was 2.6 mm. Larvae were fed ss-type rotifers from dl-d28ph, Artemia nauplii from 0–08ph, and artificial diets (52–48% protein) from d24-d38ph. On d38ph, fish averaged 0.308 g and 22.2 mm standard length. Survival (from d2ph) was 14.3%, with a total of 36,900 post-metamorphic juveniles produced. On d97ph, 1,390 hatchery-reared juveniles (avg. wt. = 10.5 g) were stocked into two 14.5-m³ recirculating seawater tanks (695 fish/tank; 48 fish/m³) and fed a 56% protein pellet. After 168 d, fish averaged 140.8 g, with a survival rate of 97.8% and a feed conversion ratio (dry wt./wet wt.) of 1.2. These preliminary results reveal the mutton snapper to be a prime, new candidate species for commercial cultivation.     

Host/parasite relationship in the in vitro infection of rat gliocytes by Neospora caninum: evaluation of cell respiration

Neospora caninum is a protozoon that causes abortion in cattle and neuromuscular lesions in dogs, with the formation of cysts mainly in the central nervous system. Since N. caninum is an intracellular parasite with tropism for the cells of nervous system, this study evaluated the respiratory metabolism of glial cells infected by this* parasite. Glial cultures obtained from the cerebral cortex of newborn rats were kept in DMEM enriched with 10% fetal bovine serum, 1 mM pyruvic acid and 2 mM of L-glutamine. They were infected at a ratio of approximately 1:1 (cell/parasite). Oxygen consumption was evaluated by polarography in the non infected and N. caninum infected groups, 24 and 72 h following infection. Glial cell respiration after 24 and 72 h was 307.2 +/- 34.7 and 308.9 +/- 64.1 microL of oxygen per mug of total protein per minute, and 566.2 +/- 54.6 and 579 +/- 117.5 microL O2/microg of total protein/minute in the control and infected groups, respectively. These results show that N. caninum does not interfere with glial respiration in vitro.