Alternative low-cost process for large-scale production of Bacillus thuringiensis in a simple and novel culture system

Abstract

An industrial-scale, profitable method for production of the most widely used bioinsecticide, Bacillus thuringiensis (Bt), is challenging because of its widespread application. The aim of this study is to present a strategy to develop a low-cost, large-scale bioprocess to produce Bt H14.

This study was first focused on the design of a culture medium composed of economical and available components, such as glycerol and lysed Saccharomyces cerevisiae. The production goal of 1200 ITU was achieved using a medium composed of 20:20 g L^?1of glycerol:lysed yeast in batch cultures. Efforts were subsequently focused on the design of an appropriate culture system, and an original two-stage culture system was proposed. First, yeast (the primary component of the culture medium) are cultivated using a minimal mineral medium and lysed, and in the second stage, Bt is cultivated in the same bioreactor using the lysed yeasts as culture medium (supplemented with a feeding pulse of 10 g L^?1 glycerol). This system was called fed batch one pot (FOP). A new inoculation strategy is also presented in this study, since these Bt cultures were inoculated directly with heat pre-treated spores instead of vegetative bacteria to facilitate the bioprocess. This study was developed from the laboratory to production-scale bioreactors (measuring from 500 mL to 2500 L), and the efficiency of the proposed strategy was evident in LD₅₀ tests results, achieving 1796 ITU in large-scale processes. Both the use of non-conventional sources and the process development for biomass production are important for cost-effective production of Bt-based insecticides in mosquito control projects.     

Volatile carbonylic compounds in downtown Santiago, Chile

Formaldehyde, acetaldehyde, acetone, propanal, butanal, 2-butenal, 3-methylbutanal, hexanal, benzaldehyde, 2-methylbenzaldehyde, and 2,5-dimethylbenzaldehyde were measured during six spring days at downtown Santiago de Chile. Measurements were performed 24h/day and averaged over three hour periods. The averages of the maxima (ppbv) were, formaldehyde: 3.9+/-1.4; butanal: 3.3+/-3.4; acetaldehyde: 3.0+/-0.9; acetone: 2.4+/-1.0; 2-butenal: 0.56+/-0.52; propanal: 0.46+/-0.21; benzaldehyde: 0.34+/-0.3; 3-butanal: 0.11+/-0.05; hexanal: 0.11+/-0.08; 2-methylbenzaldehyde: 0.08+/-0.05; 2,5-dimethylbenzaldehyde: 0.05+/-0.03. Aliphatic aldehydes (C1-C3) are strongly correlated among them and weakly with primary (toluene) and secondary (ozone plus nitrogen dioxide or PAN) pollutants. In particular, the correlation between acetaldehyde and propanal values remains even if diurnal and nocturnal data are considered separately, indicating similar sources. All these aldehydes present maxima values in the morning (9-12h) and minima at night (0-3h). The best correlation is observed when butanal and 2-butenal data are considered (r=0.99, butanal/2-butenal=6.2). These compounds present maxima values during the 3-6h period, with minima values in the 0-3h period. These data imply a strong pre-dawn emission. Other aldehydes show different daily profiles, suggesting unrelated origins. Formaldehyde is the aldehyde whose concentration values best correlate with the levels of oxidants. The contribution of primary emissions and photochemical processes to formaldehyde concentrations were estimated by using a multiple regression. This treatment indicates that (32+/-16)% of measured values arise from direct emissions, while (79+/-23)% is attributable to secondary formation.     

Artisanal fishers’ ethnobotany: from plant diversity use to agrobiodiversity management

Artisanal fishers interact with plants in many ways, and with different intensities. In spite of being characterized by fisheries, Caiçaras use plants with varied degrees of management, since the less intensive management actions, extraction of forest products, until intensively management actions through the cultivation of the agrobiodiversity. This study presents the results of different research projects and includes the North and the South regions of São Paulo state coast. The objective is to characterize the diversity of extracted and cultivated species by traditional fishers at Southeastern region of Brazilian Atlantic forest. Ethnobotanical surveys revealed a high diversity of plants known and used, ranging up to a richness of 272 species known in two communities for the South coast, most of them being native. However, depending on the management intensity, this diversity can be rather particularized: we found 68 varieties of cassava (Manihot esculenta Crantz) at North region and 58 varieties at the South region, with exclusive vernacular names. Through an ethnobotanical perspective, we compare and discuss the different dynamic systems underlying the relationships between fishers and plants. The diversity of cultivated plants is analyzed in depth, focusing on the historical importance of agriculture among people who are currently characterized as fishers. A schematic model is proposed to explain the dynamic systems operating in the relationships between fishers and plants.     

A Gradient Study of 34 Elements in the Vicinity of a Copper-Nickel Smelter in the Kola Peninsula

Concentrations of 34 elements determined by ICP mass spectrometry were studied in surface soil and vegetation along a north–south gradient through the Pechenganickel smelter complex in Kola peninsula, northern Russia. Strong influence from the smelter was evident for Fe, Co, Ni, and Cu, mainly associated with dry deposition of large particles. Also for As, Se, Mo, Sb, Te, Bi, and Pb the smelter or associated sources appeared to be distinct contributors of contamination consisting presumably of smaller particles. Significant but less distinct effects leading to enhanced concentration levels were observed for P, S, V, Cr, Zn, and Tl. In the case of Mn, Rb, Sr, Cs, and Ba the concentrations in vegetation were generally lower near the source, which may be due to cation exchange with protons or heavy metal cations in the soil and subsequent leaching from the root zone. For Li, Be, B, Na, Mg, Al, Ca, Y, Cd, La, Th, and U no particular influence from the smelter complex was observed. Some characteristic differences observed in element concentrations in different plant species and between different years of Pinus sylvestris needles are discussed. The high concentrations observed for many trace elements in the humus horizon indicates that it acts as an active biogeochemical barrier against downward transport of these elements.     

Liquid chromatography-hydride generation-atomic absorption spectrometry for the speciation of tin in seafoods

Liquid chromatography with hydride generation atomic absorption spectrometry as the detection system was applied to the separation and determination of inorganic tin, tributyltin, dibutyltin, monobutyltin, diphenyltin and monophenyltin. A reversed phase C18 column and a methanol/water/acetic acid (70:27:3, v/v/v) mixture containing 0.05%(v/v) triethylamine and 0.1%(w/v) tropolone as the mobile phase (isocratic elution) were used. Extraction of organotins from the samples was carried out using methanol containing 0.05%(w/v) tropolone, a process that was repeated twice. The supernatants were shaken with water and dichloromethane in a separating funnel and the organic phase was collected and evaporated to dryness. When the method was applied to the speciation of tin in fresh and canned mussels, no organotins above the detection limits were identified in any of the samples, inorganic tin being the only species detected. The reliability of the procedure was checked by analyzing the total tin content of the samples by electrothermal atomic absorption spectrometry and by speciation of tin in a certified reference material, mussel tissue (CRM 477). The method can be used for environmental monitoring of organotins contaminated samples.     

Analysing countries’ contribution to climate change: scientific and policy-related choices

This paper evaluates the influence of different policy-related and scientific choices on the calculated regional contributions to global climate change (the “Brazilian Proposal”). Policy-related choices include the time period of emissions, the mix of greenhouse gases and different indicators of climate change impacts. The scientific choices include historical emissions and model representations of the climate system. We generated and compared results of several simple climate models. We find that the relative contributions of different nations to global climate change—from emissions of greenhouse gases alone—are quite robust, despite the varying model complexity and differences in calculated absolute changes. For the default calculations, the average calculated contributions to the global mean surface temperature increase in 2000 are about 40% from OECD, 14% from Eastern Europe and Former Soviet Union, 24% from Asia and 22% from Africa and Latin America. Policy-related choices, such as time period of emissions, climate change indicator and gas mix generally have larger influence on the results than scientific choices. More specifically, choosing a later attribution start date (1990 instead of 1890) for historical emissions, decreases the contributions of regions that started emitting early, such as the OECD countries by 6 percentage points, whereas it increases the contribution of late emitters such as Asia by 8 percentage points. However, only including the fossil CO₂ emissions instead of the emissions of all Kyoto gases (fossil and land use change), increases the OECD contributions by 21 percentage points and decreases the contribution of Asia by 14 percentage points.     

Antibodies Against Hepatitis E Virus (HEV) in European Moose and White-Tailed Deer in Finland

Food and Environmental Virology - The main animal reservoirs of zoonotic hepatitis E virus (HEV) are domestic pigs and wild boars, but HEV also infects cervids. In this study, we estimated the...     

Differential ingestion of zooplankton by four species of bivalves (Mollusca) in the Mali Ston Bay,Croatia

This study provides information about differences in composition of ingested zooplankton amongst bivalve species coexisting in the same area in a period from May 2009 to December 2010. The study was conducted at the Mali Ston Bay (42°51′ N, 17°40′ E)—the most important bivalve aquaculture area in the eastern Adriatic Sea. Stomach content analysis was performed on cultured species—Ostrea edulis and Mytilus galloprovincialis, and commercially important bivalve species from their natural environment—Modiolus barbatus and Arca noae. Results confirmed carnivory in bivalves, both from natural and cultured populations, but cultured species had higher numbers of zooplankters than those living on the seabed. The most abundant taxa were bivalve larvae, followed by tintinnids, copepods, unidentified eggs and gastropod larvae. Recorded numbers of bivalve larvae in M. galloprovincialis stomach were the highest so far reported and show that mussels impact the availability of natural spat.     

The effect of humic substances on Cu migration in the soil profile

Humic substances (HS) are widely used for diverse purposes. The effect of HS on the metal’s status in contaminated soils is contradictory. The aim of this work was to investigate the Cu migration in soils treated with HS. A model field experiment with the addition of Cu (1.243?mg?Cu/kg) and HS Extra® (potassium humate) was performed. The Cu addition resulted in acidification (by 0.7 pH) after 3 months. The major part of the added Cu remained in the upper 7-cm-thick soil layer; 4% reached the lower soil layer, while only 0.1% were removed beyond the profile. The addition of HS mitigated soil acidification increased the content of Cu bound to solid-phase organic substances and abruptly reduced the Cu activity in the soil liquid phase. Simultaneously, the HS addition increased the water-soluble organic substances (WSOS) by four times, including those in the hydrophilic and hydrophobic fractions, resulting in a twofold increase in the content of soluble Cu. Copper complexes with hydrophilic WSOS mainly reached lysimeters, and hydrophobic organic substances were absorbed by the soil. The HS addition to a slightly acidic soil can accelerate the migration of Cu to adjacent environments.