RESUMO
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.
Assuntos
Granulovirus , Larva , Spodoptera , Animais , Spodoptera/virologia , Granulovirus/genética , Larva/virologiaRESUMO
This study aimed to investigating the possible interference caused by glass test tubes on the quantification of bacterial adhesion to hydrocarbons by the MATH test. The adhesion of four bacteria to hexadecane and to glass test tubes was evaluated employing different suspending polar phases. The role of the ionic strength of the polar phase regarding adhesion to glassware was investigated. Within the conditions studied, Gram-positive bacteria adhered to both the test tube and the hydrocarbon regardless of the polar phase employed; meanwhile, Escherichia coli ATCC 25922 did not attach to either one. The capacity of the studied microorganisms to adhere to glassware was associated with their electron-donor properties. The ionic strength of the suspending media altered the patterns of adhesion to glass in a strain-specific manner by defining the magnitude of electrostatic repulsion observed between bacteria and the glass surface. This research demonstrated that glass test tubes may interact with suspended bacterial cells during the MATH test under specific conditions, which may lead to overestimating the percentage of adhesion to hydrocarbons and, thus, to erroneous values of cell surface hydrophobicity.
Assuntos
Aderência Bacteriana , Vidro , Vidro/química , Escherichia coli , Alcanos/química , Concentração Osmolar , Interações Hidrofóbicas e Hidrofílicas , Hidrocarbonetos/metabolismo , Bactérias Gram-Positivas/isolamento & purificaçãoRESUMO
This study aimed to evaluate the effectiveness of pH control, water activity (Aw), and the addition of lactic acid bacteria (LAB) on the proliferation of Escherichia coli in the curd during the manufacturing of Chihuahua cheese. Milk proved to be an excellent culture medium for E. coli, allowing it to develop at concentrations up to 109 cfu/g. However, the presence of LAB, the pH control, Aw, and especially the use of the Cheddarization process during the Chihuahua cheese production proved to be important obstacles that inhibited the proliferation of E. coli under the conditions studied. Moreover, reducing the water activity of the curd as quickly as possible is presented as the most powerful tool to inhibit the development of E. coli during the Chihuahua cheese-making process.
RESUMO
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.
Assuntos
Inseticidas , Nucleopoliedrovírus , Animais , Spodoptera , Raios Ultravioleta , Temperatura , Zea mays , Cápsulas , Controle Biológico de Vetores , LarvaRESUMO
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.
RESUMO
Bacterial cell surface hydrophobicity is a relevant property in determining the ability of bacteria to adhere to inert surfaces. This property has been measured using the microbial adhesion to hydrocarbon (MATH) test. Several reports in the literature establish the percentage of adhesion to hydrocarbons (PoAtH) value produced by the MATH test for a broad variety of bacteria. Discrepancies in PoAtH values reported for the same strain of a specific microorganism suggest that some method-induced variation may exist, as different research teams employ different versions of the assay. The objective of the present study was to compare the performance of different versions of the MATH test as reported in the literature, to quantify the magnitude of the method-induced variation on PoAtH values. The study demonstrated that PoAtH values are influenced twice as much by variations in the employed assay than by actual differences in cell surface composition or architecture. The two L. reuteri strains studied responded differently to changes in assay conditions showing 40 and 70% method-dependent variation for strain ATCC 53609 and 55730, respectively. These results highlight the need to properly standardize the MATH test to enable comparison of PoAtH values produced by independent research teams.
Assuntos
Aderência Bacteriana , Bioensaio/métodos , Membrana Celular/metabolismo , Interações Hidrofóbicas e Hidrofílicas , Bactérias/crescimento & desenvolvimento , Bactérias/metabolismo , Humanos , Hidrocarbonetos/metabolismo , Limosilactobacillus reuteri , TemperaturaRESUMO
Insects are considered as alternative sources of chitosan; however, studies about the functional film-forming properties of insect chitosan are scarce. Insect chitosan films were made from Tenebrio molitor and Brachystola magna and were compared with commercial chitosan of different molecular weights (Mw). Mechanical properties (tensile strength, TS; elastic modulus, EM; elongation at break, %E), water vapor permeability (WVP) and physicochemical properties were characterized. The film properties of both commercial and insect chitosan were affected by Mw. Commercial chitosan films showed that at lower Mw, the TS (from 59 to 48 MPa) and EM (from 1471 to 1286 MPa) decreased; whereas WVP (from 2.9 × 10-11 to 3.4 × 10-11 g m-1s-1Pa-1), % E (from 38 to 41%) and solubility (from 30 to 33%) increased. Chitosan insect films showed lower TS and EM, and higher WPV, %E and solubility than commercial films. SEM revealed that chitosan insect films had lower porosity than commercial films. FTIR and X-ray diffraction showed not difference between insect and commercial chitosan films. These results showed that T. molitor and B. magna chitosan films could be used as a packaging material in several food products.
Assuntos
Quitosana/química , Membranas Artificiais , Tenebrio/química , Animais , Fenômenos Químicos , Fenômenos Mecânicos , Peso Molecular , Reologia , Solubilidade , Análise Espectral , VaporRESUMO
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.
Assuntos
Abelhas/microbiologia , Enterococcaceae/fisiologia , Filogenia , Animais , Larva/microbiologia , México , PrevalênciaRESUMO
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.
Assuntos
Quimosina/genética , Quimosina/metabolismo , Pichia/genética , Análise de Sequência de DNA/métodos , Animais , Bovinos , Códon , Meios de Cultura/química , Genes Sintéticos , Proteínas Recombinantes/metabolismo , Transformação GenéticaRESUMO
Organic strawberry fruits (Cv. 'Albion') were harvested at six different ripening stages and evaluated for physical and chemical parameters. Biometrical characteristics and moisture content did not change significantly during ripening. Total soluble solids, pH and colour development increased while titratable acidity and fruit firmness decreased 14.7% and 91%, respectively. Fructose, glucose, and sucrose followed similar tendencies. Final contents of these sugars were 2323.4, 1988.5, and 1578.4 mg/100 g. Citric, malic, and ascorbic acids followed a descending, irregular, and increasing tendency during ripening, respectively. Final contents of these acids were 822.8, 245.8, and 78.1 mg/100 g. Total anthocyanins content (TAC) increased during ripening, while the opposite was observed for total phenolic content (TPC). TAC and TPC in ripe fruit were 56.4 mg/100g and 196 mg gallic acid equivalents (GAE)/100 g. Twenty eight phenolic compounds, mainly glycosides, were identified and quantified by HPLC-DAD-MS analysis. The concentration of these compounds was ripening dependent.