Genome Mining for Antimicrobial Compounds in Wild Marine Animals-Associated Enterococci

New ecosystems are being actively mined for new bioactive compounds. Because of the large amount of unexplored biodiversity, bacteria from marine environments are especially promising. Further, host-associated microbes are of special interest because of their low toxicity and compatibility with host health. Here, we identified and characterized biosynthetic gene clusters encoding antimicrobial compounds in host-associated enterococci recovered from fecal samples of wild marine animals remote from human-affected ecosystems. Putative biosynthetic gene clusters in the genomes of 22 Enterococcus strains of marine origin were predicted using antiSMASH5 and Bagel4 bioinformatic software. At least one gene cluster encoding a putative bioactive compound precursor was identified in each genome. Collectively, 73 putative antimicrobial compounds were identified, including 61 bacteriocins (83.56%), 10 terpenes (13.70%), and 2 (2.74%) related to putative nonribosomal peptides (NRPs). Two of the species studied, Enterococcus avium and Enterococcus mundtti, are rare causes of human disease and were found to lack any known pathogenic determinants but yet possessed bacteriocin biosynthetic genes, suggesting possible additional utility as probiotics. Wild marine animal-associated enterococci from human-remote ecosystems provide a potentially rich source for new antimicrobial compounds of therapeutic and industrial value and potential probiotic application.     

ACCUMULATION OF SILICON IN CYNODON DACTYLON X C. TRANSVAALENSIS AND POA TRIVIALIS USED AS AN OVERSEED GRASS

Calcium silicate has been used to supply silicon (Si) as a plant nutrient for enhancing crop production and the suppression of turf diseases. Research was initiated to determine the effect of calcium silicate on silicon uptake, turf quality, and its residual activity in providing plant available Si in two turfgrass systems. During 2005 and 2006, calcium silicate (CaSi, 12% Si) was applied as a topdressing at eight rates ranging from 0 to 342 kg m^?2 to a ‘Tifdwarf’ (Cynodon dactylon × C. transvaalensis Burtt Davy) green in the spring and to the same plots overseeded with ‘Wintergreen’ rough bluegrass (Poa trivialis L.) in the fall. At initiation of the study, dolomitic limestone and magnesium sulfate (MgSO₄) was applied to equilibrate calcium across all treatments based on the highest rate of CaSi. Every 28 days, treatments were rated for turf quality and then harvested to determine Si accumulation. One month after applying CaSi in 2005, Si concentration in the leaf tissue of Tifdwarf had increased linearly with increasing rates. This response continued through the months of May, June, July, August, and October with a 9%, 21%, 40%, 22% and 21% increase in Si concentration in the leaf tissue between the control and the 342 kg m^?2 rate, respectively. Although the rate response remained, there was an overall drop in average percent Si accumulation in leaf tissue across the treatments beginning three months after the first application. Similar responses were noted with the overseeded rough bluegrass with a 46%, 85%, 69%, 111% and 58% increase in Si concentration in the leaf tissue between the control and the 342 kg m^?2 rate, respectively. In contrast to the bermudagrass, over the five month period following application, the residual activity of Si in rough bluegrass remained high, suggesting that rough bluegrass might be a better Si accumulator in comparison to Tifdwarf bermudagrass. The second run of the experiment (2006–2007) and the residual trial had the same response with Si concentration in the leaf tissue increasing as the amount of CaSi applied increased. Although turfgrass quality was not well correlated to CaSi rate for Tifdwarf bermudagrass, there was an increase in quality ratings for rough bluegrass from December 2005 to April 2006.     

The Effect of Calcium Sprays and Fruit Thinning on Bitter Pit Incidence and Calcium Content in ‘Honeycrisp’ Apple

‘Honeycrisp’ is a recently released cultivar of apple (Malus x domestica) developed by the University of Minnesota, now in high demand by growers and consumers. This cultivar is susceptible to bitter pit, especially when the trees are young and the fruit load is low. The effects of calcium (Ca) sprays, fruit thinning, vegetative growth, fruit and leaf nutrient concentration on bitter pit incidence, and two sampling methods for nutrient analysis were studied. Bitter pit incidence was associated with several variables, including Ca and phosphorus (P) concentrations in leaves and fruits, magnesium (Mg) concentration in leaves, and yield parameters. Fruit Ca concentration and content for the growing season were modeled, which allowed the determinations of Ca levels associated with bitter pit incidence. Our data suggest that plug analysis was less sensitive than fruit-segment analysis for finding differences in Ca concentrations.     

INTERRELATION BETWEEN FRUIT MINERAL CONTENT AND PRE-HARVEST CALCIUM TREATMENTS ON ‘GOLDEN SMOOTHEE’ APPLE QUALITY

The effects of different pre-harvest calcium treatments and their interrelation with other nutrients in apples (Malus domestica Borkh, ‘Golden Smoothee’) on quality attributes at harvest and after cold storage and on relation between calcium (Ca) and other nutrients were investigated. Calcium sprays were applied during three-seasons beginning 60 days after full bloom (DAFB), with 6 or 12 applications in the second part of the growing season. It was confirmed that calcium treatments generally resulted in increased flesh firmness at harvest but not sufficient to maintain during storage. Treatments reduced bitter pit and lenticel blotch pit after cold storage, but these disorders were not totally eliminated. The treatments did not influence evolution of titratable acidity and soluble solids. Total fruit calcium increased with number of applications, but this increase was not proportional with number of applications. The beneficial effect of calcium treatments was reflected in the nitrogen/Calcium (N/Ca), potassium/calcium (K/Ca), and (K+Mg)/Ca ratios.     

Enhancement of natural disease resistance in potatoes by chemicals

The mechanism involved in systemic acquired resistance (SAR) can be non-specifically induced in susceptible plants. In response to pathogens, plants' natural defence mechanisms include the production of lignin and phytoalexins and the induction of plant enzymes. The aim of this research was to study the induction of SAR mediated by the chemical activator DL-3-aminobutyric acid (BABA) and the fungicide fosetyl-aluminium in potato cultivars with different levels of resistance against Phytophthora infestans (Mont) de Bary. To study the chemical induction of the resistance, the foliage of several potato cultivars was sprayed with BABA, fosetyl-aluminium or water (as a control treatment). After 3 days the foliage was inoculated with P. infestans. Seven days after inoculation, development of disease symptoms in the foliage was assessed. In postharvest tuber samples, evidence for enhancement of the defence response was evaluated by measuring the protein content of several hydrolytic enzymes as well as the phenol and phytoalexin content. The highest level of protection against late blight was observed when the chemicals were applied at early stages of crop development. An increase in resistance to late blight was also detected in tubers after harvest. There was also an increase in the protein level of beta-1,3-glucanase and aspartic protease as well as in the phenol and phytoalexin content of potato tuber discs obtained from postharvest tubers of treated plants. Thus the protective effect seemed to persist throughout the whole crop cycle. This treatment may offer the possibility of controlling both foliage and tuber blight and could have a major impact in reducing over-winter survival of P. infestans in tubers.     

Genetics of Melon Yellows Virus Resistance Derived from Cucumis melo ssp. agrestis

The Melon yellows virus (MYV), a whitefly-transmitted closterovirus, is one of the major pathogens causing crop losses in protected melons in southeastern Spain. An accession of the wild Asiatic Cucumis melo ssp. agrestis (Cma) shows resistance to MYV infection. Results indicate the participation of two resistance mechanisms in this source: firstly, an antixenotic reaction against Trialeurodes vaporariorum, the disease vector, and secondly, resistance to the virus. The combined effect of these two mechanisms confers Cma a higher level of resistance, expressed as a delayed and milder infection.The genetics of resistance to the Melon yellows closterovirus have been studied in two families derived from Cma. As under natural infection conditions, the effect of antixenosis and virus resistance cannot be distinguished, a biometrical model that permits separation of the two resistance mechanisms operating in the same resistant source, has been proposed to determine genetic control of MYV resistance.The genetic analysis has been conducted by fitting the disease progress curves of each generation to the biometrical model instead of fitting the final disease ratios. The scoring of disease incidence over time allows for the comparison of data from assays conducted in different conditions (2 years/4 transplanting dates), thus reinforcing the analysis.The results agree with a simple control of the resistance to MYV derived from Cma, with incomplete penetrance of the gene and partial dominance of resistance. The effect of antixenosis on the spread of this plant virus is highly significant in Cma, but not in segregant generations.Since there do not exist crossability barriers between this accession and the cultivated melon, Cma could be readily used in breeding programmes to obtain melon varieties resistant to MYV.     

Effect of the density on the growth and survival of the Caribbean scallops Argopecten nucleus and Nodipecten nodosus in suspended culture

Juveniles of the Caribbean scallops Argopecten nucleus and Nodipecten nodosus were suspended in pearl nets at densities of 25%, 30% and 50% coverage of the bottom of the net (BC) during days. Additionally, adults of A. nucleus (>35 mm) were placed in lantern nets at densities of 40%, 50% and 65% BC during days. Stocking density affected the growth of animals, with higher growth rates obtained at the lower densities. Although the survival of the two scallops was not influenced at the densities studied, the 15-day net changes, and culture in water of <28 °C produced better survival of A. nucleus and N. nodosus respectively. Predators of the families Cymatiidae, Grapsidae, Portunidae, Majidae and Xanthidae were found in the culture nets of juvenile A. nucleus , but during the adult phase they were absent. Predators exhibited a dense-independent behaviour in relation to scallops. There was not any association between predator frequency and survival of A. nucleus .     

Evaluation of the Microalgae Paste Viability Produced in a Mollusk Hatchery in Southern Brazil

The present study was conducted to define a methodology to produce and store small‐scale microalgae paste to be used in a mollusk hatchery. Microalgae were cultured in 500 L fiberglass tanks, under temperature of 20 ± 2 C, Guillard f/2 culture medium, and continuous light intensity of 203–226 μmol photons/m²/sec. Cultures were centrifuged at 2000 g at the exponential growth phase. Microalgae cell quality after centrifugation and during storage was determined by analyses with Evan’s blue stain and by counting the number of total marine bacteria. Treatments with and without additive were applied to the microalgae paste produced, which was distributed into 100 mL plastic containers, capped, and stored under refrigeration at 4 ± 1 C. Results indicated that in the Chaetoceros muelleri paste, centrifugation did not damage the cells and the number of total marine bacteria reduced significantly from 2.9 × 10⁶ to 8.3 × 10⁵ colony‐forming units per milliliter. Chaetoceros muelleri and Chaetoceros calcitrans pastes stored with addition of 0.1% ascorbic acid had a shelf life shorter than 2 wk. For the treatment without additive, results with Evan’s blue stain showed that cells (99%) remained viable until the sixth week of storage for C. muelleri and seventh week of storage for Skeletonema sp. and C. calcitrans. The number of bacteria did not increase during storage for C. calcitrans and Skeletonema (P > 0.05). For C. muelleri, an increase in bacteria (P < 0.05) was observed after the sixth week of storage. This study demonstrated the feasibility to produce and store microalgae paste for a period of 2–8 wk, which allows it to be used as food source and also optimizes the use of microalgae cultured in laboratory.     

Evolution of Carotenoids,Sensory Profiles and Volatile Compounds in Microwave-Dried Fruits of Three Different Loquat Cultivars (Eriobotrya japonica Lindl.)

Plant Foods for Human Nutrition - The consumption of loquat fruits is highly appreciated for their carotenoid content and valuable sensory notes, but it is limited due to the low shelf-life. An...