Morphological, Biological, and Molecular Characterization of Type I Granuloviruses of Spodoptera frugiperda.

Granuloviruses (GVs) Betabaculovirus associated with the fall armyworm (FAW), Spodoptera frugiperda (J.E. Smith) (Lepidoptera: Noctuidae), especially those of the type I, have scarcely been studied. These GVs might be an effective alternative for the biocontrol of this insect. In this study, the native GVs SfGV-CH13 and SfGV-CH28 were isolated from FAW larvae and characterized for morphology, molecular traits, and insecticidal activity. The elapsed time between symptomatic infection of larvae and stop feeding as well as the weight of larvae before death or prior to pupation were also evaluated. Both GVs had ovoid shape and a length of 0.4 µm. They had the same DNA restriction profiles and their genome sizes were about 126 kb. The symptomatic infection with the tested GVs mainly caused flaccidity of larva body and discoloration of integument. The integument lysis was only observed in 8% of infected larvae. Infected larvae gradually stopped feeding. Overall, these symptoms are characteristic of infections caused by type I GVs, which are known as monoorganotropic or slow-killing GVs. The median lethal dose (LD50) values for SfGV-CH13 and SfGV-CH28 isolates were 5.4 × 102 and 1.1 × 103 OBs/larva, respectively. The median lethal time (LT50) ranged from 17 to 24 days. LT50 values decreased as the viral dose was increased. The elapsed time from symptomatic infection until pupation and body weight of larvae (third instar) were higher with SfGV-CH28 than SfGV-CH13. Both granulovirus isolates were able to kill the FAW larvae from the 12th day.

Foliar Application of CaCO3-Rich Industrial Residues on 'Shiraz' Vines Improves the Composition of Phenolic Compounds in Grapes and Aged Wine.

The quality of wine grapes and wine depends on their content of phenolic compounds. Under commercial conditions, the phenolic maturity of grapes is mostly achieved by applying abscisic acid analogues. Some Ca forms represent a cost-effective alternative for these compounds. In this study, 'Shiraz' vines (veraison of 90%) were sprayed with CaCO3-rich residues from the cement industry (4.26 g of Ca per L). Fruit from treated and untreated vines was harvested 45 days after CaCO3 spraying and evaluated for quality. The fruit was vinified, and the obtained wines were bottled and stored in darkness for 15 months at 20 °C. Wines were evaluated for quality after storage. The evaluation of grape and wine quality included the content of phenolic compounds and antioxidant capacity. The treatment with CaCO3 did not affect the ripening rate of grapes. However, the treatment improved the fruit yield as well as the color development, the content of phenolic compounds, and antioxidant capacity of grapes and wine. The treatment favored especially the accumulation of malvidin-3-O-glucoside, pelargonidin-3-O-glucoside, caftaric acid, caffeic acid, trans-cinnamic acid, quercetin, catechin, epicatechin, resveratrol, and the procyanidins B1 and B2. Wine made with treated fruit was of higher quality than that of control fruit.

Effect of High Temperature and UV Radiation on the Insecticidal Capacity of a Spodoptera frugiperda Nucleopolyhedrovirus Microencapsulated in a Matrix Based on Oxidized Corn Starch.

A multiple nucleopolyhedrovirus native isolate (SfCH32) of Spodoptera frugiperda (J.E. Smith) (Lepidoptera: Noctuidae) was encapsulated by spray-drying in a matrix based on oxidized corn starch without and with a fluorescent brightener. The microcapsules were exposed to UV radiation (365 nm) for 0, 2, 4, and 8 h at 25 °C or temperatures of 35, 40, and 45 °C for 8 h. The data obtained with temperatures 35, 40, and 45 °C were contrasted with those obtained at 25 °C. The microcapsules were evaluated for size, shape, and insecticidal capacity against third instar S. frugiperda larvae under laboratory conditions. The 82-84.2% of the encapsulating matrix, in a dry-weight basis, was recovered as NPV microcapsules of heterogeneous shape and size. The exposure to UV radiation and temperatures reduced significantly the insecticidal capacity of tested viruses; however, such capacity was higher for microencapsulated than for non-microencapsulated viruses. The non-encapsulated virus that had been exposed to 45 °C or maintained at UV radiation for 8 h showed the lowest insecticidal activity at 5th day post-inoculation, with a larvae mortality of 25.3 and 16%, respectively. The fluorescent brightener increased significantly the insecticidal capacity of encapsulated and non-encapsulated viruses, causing a mortality of 100% at that time point, and decreased the median lethal time independently of the incubation temperature and exposure time to radiation. The findings suggested that an encapsulating matrix based on oxidized corn starch might protect the insecticidal capacity of NPV under field conditions.

In vitro biocontrol of Fusarium oxysporum with antagonistic microorganisms and In vivo effect on Solanum lycopersicum L.

The objective was to evaluate the effects of In vitro biocontrol of Trichoderma spp. and Bacillus spp. against Fusarium oxysporum and its in vivo effect on Solanum lycopersicum plants. The antagonistic effect in vitro of microcapsules of Trichoderma spp. and Bacillus spp. against F. oxysporum, was evaluated by inhibition and antagonism classification, in the in vivo part growth variables in plant and fruit were measured. Trichoderma asperellum showed inhibition of 84.7% and class 1 and Bacillus spp., obtained an inhibition of 83.7% with a positive classification. The canonical correspondence analysis of the fruit, the T1 treatment indicates that the variables that highlight the effect of the treatment are the firmness of the fruit and the Brix. The canonical correspondence analysis of the plant, the height presents a significant difference (P < 0.05), highlighting B. subtilis, Bacillus liquefasciens, T. harzianum and T. asperellum + Bacillus amiloliquefasciens, with 1.05 and 1.1 m compared to the control of 74 cm, also highlighted the number of total fruits in the T. harzianum treatment with an average of 22.6 fruits and root weight in B. subtilis with 37.12 g. Strains of Trichoderma spp. and Bacillus spp. successfully inhibited F. oxysporum also induced growth in the tomato plant.

Growth Promotion of Phaseolus vulgaris and Arabidopsis thaliana Seedlings by Streptomycetes Volatile Compounds

Streptomyces are recognized as antipathogenic agents and plant-growth-promoting rhizobacteria. The objective of this study was to evaluate the capacities of four antifungal Streptomyces strains to: produce the substances that are involved in plant growth; solubilize phosphates; and fix nitrogen. The effects of the volatile organic compounds (VOCs) that are emitted by these strains on the growth promotion of Arabidopsis thaliana and Phaseolus vulgaris L. (var. Pinto Saltillo) seedlings were also tested. All of the Streptomyces strains produced indole-3-acetic acid (IAA) (10.0 mg/L to 77.5 mg/L) and solubilized phosphates, but they did not fix nitrogen. In vitro assays showed that the VOCs from Streptomyces increased the shoot fresh weights (89-399%) and the root fresh weights (94-300%) in A. thaliana seedlings; however, these effects were less evident in P. vulgaris. In situ experiments showed that all the Streptomyces strains increased the shoot fresh weight (11.64-43.92%), the shoot length (11.39-29.01%), the root fresh weight (80.11-140.90%), the root length (40.06-59.01%), the hypocotyl diameter (up to 6.35%), and the chlorophyll content (up to 10.0%) in P. vulgaris seedlings. 3-Methyl-2-butanol had the highest effect among the ten pure VOCs on the growth promotion of A. thaliana seedlings. The tested Streptomyces strains favored biomass accumulation in A. thaliana and P. vulgaris seedlings.

Gastrointestinal metabolism and bioaccessibility of selected anthocyanins isolated from commonly consumed fruits.

There is uncertainty about the identity of digestive metabolites of anthocyanins because many are naturally present in foods and/or are formed from other phenolic compounds during the digestive process. Studies using pure anthocyanins are needed to clarify this uncertainty. In this study, selected anthocyanins were purified from common fruits and individually subjected to gastric and small intestinal digestion in vitro to determine their stability, metabolites generated and bioaccessibility. Anthocyanins were highly stable during the gastric phase of simulated digestion (p > 0.05). The recovery of anthocyanins decreased during the small intestinal phase of digestion (p < 0.05). Stability was dependent on anthocyanidin structure and type of glycation (p < 0.05). Gastric and gastrointestinal phases mainly contained anthocyanins as bioaccessible flavylium cations and chalcones. Expected anthocyanin metabolites (i.e., phenolic acids and phoroglucinaldehyde) were not detected in chyme. Deglycation of anthocyanins during simulated digestion was quite limited and the bioaccessibility of intact anthocyanins was very low (0.07-2.21%).

Physicochemical, Structural, Thermal and Rheological Properties of Flour and Starch Isolated from Avocado Seeds of Landrace and Hass Cultivars.

The objective of this study was to obtain and characterize flours and starches from the avocado seeds of Hass and landrace cultivars. The morphological, physical-chemical, structural, thermal and rheological characteristics were evaluated. The flour yield of the Hass and landrace cultivars was 41.56 to 46.86% (w/w), while for starch, it was 35.47 to 39.57% (w/w) (cv. Hass and landrace, respectively). Scanning electron microscopy (SEM) revealed the presence of oval starch granules and other particles in flour, in contrast to flours, starches showed lower ash, proteins and lipids content. However, the amylose content was higher in starches (42.25-48.2%). Flours showed a higher gelatinization temperature (Tp = 73.17-73.62 °C), and their starches presented greater gelatinization enthalpy (∆Hgel = 11.82-13.43 J/g). All samples showed a B-type diffraction pattern, and the crystallinity was higher in the flours. The rheological analysis (flow curves and viscoelastic tests) evidenced a pseudoplastic (n = 0.28-0.36) behavior in all samples analyzed, but the consistency index (k) was higher in starches. In general, the flours and starches from avocado seeds presented interesting proximal, thermal and functional properties for possible application in food systems, and these findings could contribute to the revaluation of this by-product.

Virulence variations between clonal complexes of Melisococcus plutonius and the possible causes.

Melissococcus plutonius is a pathogenic bacterium that affects honeybee brood triggering colony collapse in severe cases. The bacterium causes a European foulbrood (EFB) disease in the honeybee populations, impacting beekeeping and agricultural industries. The pathogenesis, epidemiology, and variants of M. plutonius have been studied, but the virulence factors involved in larval infection are still unknown. Recently, an in-silico study suggested putative genes that might play a role in the pathogenesis of EFB. However, studies are required to determine their function as virulence factors. In addition, the few studies of clonal complexes (CCs), virulence factors, and variation in the honeybee larvae mortality have interfered with the development of more efficient control methods. The research, development, and differences in virulence between genetic variants (CCs) of M. plutonius and potential virulence factors implicated in honeybee larval mortality are discussed in this review.

Effect of cultivar on the content of selected phytochemicals in avocado peels.

The peels of ripe fruit of 'Hass' and 'Hass' type (HT) avocado cultivars were evaluated for phytochemical composition and other attributes. Peels represented from 8.78 to 14.11% of fruit weight. Their color ranged from homogeneous black to black with very small greenish spots. The oil content in the peels was low. Twelve fatty acids were identified in peel oil and the ratio of unsaturated to saturated fatty acids suggested that peel oil might contribute to human health. The phytochemical composition varied significantly with cultivar. However, many HT peels were superior than 'Hass' peel in their content of α-tocopherol, ß-sitosterol, perseitol, and cyanidin-3-glucoside, which was up to 211.67, 45.92, 337.17, and 519.27% higher in HT peels, respectively. The content of some phenolic compounds, especially procyanidin B2 and epicatechin, was significantly lower in 'Hass' than in many HT peels. Few HT peels showed a higher content of carotenoids and chlorophyll than 'Hass' peels. Lutein was the most abundant carotenoid. Chlorophyll a and b were also abundant in peels and low concentrations of chlorophyll derivatives were observed. Avocado peels are an important source of bioactive compounds, including some carotenoids, acids, sterols, and volemitol, which were observed for the first time.

Metabolomic analysis and physical attributes of ripe fruits from Mexican Creole (Persea americana var. Drymifolia) and 'Hass' avocados.

The physicochemical properties, including nutrient and bioactive compound compositions, in fruit of four creole avocados (CA) from Mexico were determined and compared with those of 'Hass' fruit. 'Hass' pulp and some CA pulps contained similar concentrations of lutein, chlorophyll a, ß-sitosterol and α-tocopherol. CA pulp contained 3.91-9.55% more oil than 'Hass'. Oil from CA pulp contained 10.10-26.79% more oleic acid than 'Hass' pulp. However, CA were small (CA = 81.40-137.15 g, 'Hass' = 188.59 g) and their pulp contents were low (CA = 39.83-84.82 g, 'Hass' = 144.14 g). CA peels were very thin, making these avocado peels edible but prone to mechanical damage. CA peels also contained higher concentrations and greater diversity of anthocyanins and glycosylated quercetin compounds than 'Hass' peels. Some CA were particularly rich in mannoheptulose and perseitol. Consumption of CA, including their peel, might result in higher intakes of some nutrients and bioactive compounds compared with 'Hass' avocados.

Prevalence, typing and phylogenetic analysis of Melissococcus plutonius strains from bee colonies of the State of Chihuahua, Mexico.

European foulbrood (EFB) caused by Melissococcus plutonius is an important bee brood disease but, in Mexico, information about this bacterium is limited. We evaluated the prevalence of typical and atypical strains in beehives of seven apicultural regions of the state of Chihuahua, Mexico. We performed MLST and phylogenetic analysis to characterize the isolates. Prevalence was highest 59%, in the region of Chihuahua, and lowest, 14%, in the regions of Cuauhtémoc and Nuevo Casas Grandes. Typical and atypical strains were identified in hives from all regions; however, in the regions of Parral, Cuauhtémoc and Aldama, the atypical strains were only detected in combination with typical strains. We obtained 81 isolates of M. plutonius and identified seven sequence types, of which three were new types. Additionally, we observed a relation between sequence type and the region where the strain was isolated. Phylogenetic analysis and multilocus sequence typing using goeBURST analysis showed that 97.5% of the isolates correspond to the Clonal Complex (CC) 12 and 2.5% to the CC3. Our work is the first molecular characterization of M. plutonius in Mexico and contributes to global information about the epidemiology of this pathogen.

Relationship between the firmness of Golden Delicious apples and the physicochemical characteristics of the fruits and their pectin during development and ripening.

Firmness at harvest determines the postharvest storability and quality of apples. The climate change has altered the physiological processes of fruits and the reliability of ripening indicators typically used to determine the harvest time, compromising fruit firmness. In this study, 'Golden Delicious' apples were harvested at several developmental/ripening stages (107, 122, 137, 152 and 167 days after full bloom, DAFB) and evaluated for physicochemical attributes, which were correlated with fruit firmness. The 167 DAFB fruit corresponded to fruit at the commercial harvest. Fruit harvested at 107 and 122 DAFB did not develop the characteristics of ripe fruit while fruit harvested after 137 DAFB ripened normally. Fruit at commercial harvest showed low firmness. The changes of fruit weight, diameter, height, tristimulus color (L* values) as well as the content of total soluble solids and moisture in fruit correlated well with fruit firmness (r values from -0.76 to -0.97). The changes of pectin content, degree of esterification, molecular weight and content of glucose and galactose in pectin showed a positive relationship with firmness (r = 0.62-0.94). The content of protein, galacturonic acid and mineral elements in pectin correlated negatively with firmness (r -0.66 to -0.99). The results demonstrated that commercial harvest was delayed 30 days, compromising fruit firmness. Some underestimated ripening indicators may help in determining the harvest time of apples.

Effects of pectin on lipid digestion and possible implications for carotenoid bioavailability during pre-absorptive stages: A review.

Pectin, an abundant polysaccharide in the human diet, has structural characteristics and functional properties that are strongly dependent on the food matrix (e.g., origin, type, cultivar/variety, ripening stage, style and intensity of processing). These polysaccharides have a strong effect on lipid digestion, which is required for the liberation of carotenoids from emulsified lipid droplets in the gastrointestinal content and for the formation of micelles, in which the carotenoids must be incorporated before absorption. Only micellarized carotenoids can be absorbed and subsequently exert protective effects on human health. The alteration of lipolysis by pectin can occur through several mechanisms; however, they have not been linked directly to carotenoid micellarization. This paper provides an overview of the effects of the properties of pectin on the ion concentration in the digestive content, the viscosity of the digestive medium, the properties of the lipid droplet surfaces and lipase activity and analyzes the impact of these events on lipid digestion and subsequent carotenoid micellarization.

Codon Optimization of the "Bos Taurus Chymosin" Gene for the Production of Recombinant Chymosin in Pichia pastoris.

Codon optimization of the Bos taurus Chymosin gene (CYM) for its expression in Pichia pastoris was performed in this study. A synthetic CYM gene was designed in silico by replacing codons rarely used by P. pastoris with equivalent nucleotide combinations that codify for the same amino acid but that are more frequently encountered in the genome of P. pastoris. A total of 332 nucleotides were modified to optimize 289 codons. The synthetic CYM gene was cloned into the expression vector pPICZαA and transformed into P. pastoris. The transformed strains were grown in artificial media supplemented with glycerol as a carbon source to increase biomass and then cultured in a similar medium replacing glycerol with methanol as a carbon source to initiate gene induction. Raw extracts of the growth media exhibited milk-clotting activity of 146.11 SU/mL. Produced recombinant chymosin showed coagulant activity from 25 to 50 °C, and within a pH range of 5-6.9, having optimum activity at 35-40 °C, and pH 5.0. These results show that codon optimization is a viable strategy to improve CYM gene expression levels in P. pastoris for the production of recombinant chymosin.