Presence of starch enhances in vitro biodegradation and biocompatibility of a gentamicin delivery formulation

The effect of α-amylase degradation on the release of gentamicin from starch-conjugated chitosan microparticles was investigated up to 60 days. Scanning electron microscopic observations showed an increase in the porosity and surface roughness of the microparticles as well as reduced diameters. This was confirmed by 67% weight loss of the microparticles in the presence of α-amylase. Over time, a highly porous matrix was obtained leading to increased permeability and increased water uptake with possible diffusion of gentamicin. Indeed, a faster release of gentamicin was observed with α-amylase. Starch-conjugated chitosan particles are non-toxic and highly biocompatible for an osteoblast (SaOs-2) and fibroblast (L929) cell line as well as adipose-derived stem cells. When differently produced starch-conjugated chitosan particles were tested, their cytotoxic effect on SaOs-2 cells was found to be dependent on the crosslinking agent and on the amount of starch used.

Who is who in anaerobic oil biodegradation?

[Excerpt] Anaerobic bioremediation is an important alternative for the common aerobic cleanup of subsurface petroleum-contaminated soil and water. Microbial communities involved in anaerobic oil biodegradation are scarcely studied, and only few mechanisms of anaerobic hydrocarbons degradation are described. In this work, microbial degradation of aliphatic hydrocarbons (AHC) was studied by using culture-dependent and culture-independent approaches. Hexadecane and hexadecene-degrading microbial communities were enriched under sulfate-reducing and methanogenic conditions. The microorganisms present in the enriched cultures were identified by 16S rRNA gene sequencing. (...)

Cyanobacteria in Iowa Waters, 2005

This monthly report from the Iowa Department of Transportation is about the water quality management of Iowa's rivers, streams and lakes.

Examining chemical compound biodegradation at low concentrations through bacterial cell proliferation.

We show proof of principle for assessing compound biodegradation at 1-2 mg C per L by measuring microbial community growth over time with direct cell counting by flow cytometry. The concept is based on the assumption that the microbial community will increase in cell number through incorporation of carbon from the added test compound into new cells in the absence of (as much as possible) other assimilable carbon. We show on pure cultures of the bacterium Pseudomonas azelaica that specific population growth can be measured with as low as 0.1 mg 2-hydroxybiphenyl per L, whereas in mixed community 1 mg 2-hydroxybiphenyl per L still supported growth. Growth was also detected with a set of fragrance compounds dosed at 1-2 mg C per L into diluted activated sludge and freshwater lake communities at starting densities of 10(4) cells per ml. Yield approximations from the observed community growth was to some extent in agreement with standard OECD biodegradation test results for all, except one of the examined compounds.

Enhanced biodegradation of hexachlorocyclohexane (HCH) in contaminated soils via inoculation with Sphingobium indicum B90A.

Soil pollution with hexachlorocyclohexane (HCH) has caused serious environmental problems. Here we describe the targeted degradation of all HCH isomers by applying the aerobic bacterium Sphingobium indicum B90A. In particular, we examined possibilities for large-scale cultivation of strain B90A, tested immobilization, storage and inoculation procedures, and determined the survival and HCH-degradation activity of inoculated cells in soil. Optimal growth of strain B90A was achieved in glucose-containing mineral medium and up to 65% culturability could be maintained after 60 days storage at 30 degrees C by mixing cells with sterile dry corncob powder. B90A biomass produced in water supplemented with sugarcane molasses and immobilized on corncob powder retained 15-20% culturability after 30 days storage at 30 degrees C, whereas full culturability was maintained when cells were stored frozen at -20 degrees C. On the contrary, cells stored on corncob degraded gamma-HCH faster than those that had been stored frozen, with between 15 and 85% of gamma-HCH disappearance in microcosms within 20 h at 30 degrees C. Soil microcosm tests at 25 degrees C confirmed complete mineralization of [(14)C]-gamma-HCH by corncob-immobilized strain B90A. Experiments conducted in small pits and at an HCH-contaminated agricultural site resulted in between 85 and 95% HCH degradation by strain B90A applied via corncob, depending on the type of HCH isomer and even at residual HCH concentrations. Up to 20% of the inoculated B90A cells survived under field conditions after 8 days and could be traced among other soil microorganisms by a combination of natural antibiotic resistance properties, unique pigmentation and PCR amplification of the linA genes. Neither the addition of corncob nor of corncob immobilized B90A did measurably change the microbial community structure as determined by T-RFLP analysis. Overall, these results indicate that on-site aerobic bioremediation of HCH exploiting the biodegradation activity of S. indicum B90A cells stored on corncob powder is a promising technology.

Sponge Mass Mortalities in a Warming Mediterranean Sea: Are Cyanobacteria-Harboring Species Worse Off?

Mass mortality events are increasing dramatically in all coastal marine environments. Determining the underlying causes of mass mortality events has proven difficult in the past because of the lack of prior quantitative data on populations and environmental variables. Four-year surveys of two shallow-water sponge species, Ircinia fasciculata and Sarcotragus spinosulum, were carried out in the western Mediterranean Sea. These surveys provided evidence of two severe sponge die-offs (total mortality ranging from 80 to 95% of specimens) occurring in the summers of 2008 and 2009. These events primarily affected I. fasciculata, which hosts both phototrophic and heterotrophic microsymbionts, while they did not affect S. spinosulum, which harbors only heterotrophic bacteria. We observed a significant positive correlation between the percentage of injured I. fasciculata specimens and exposure time to elevated temperature conditions in all populations, suggesting a key role of temperature in triggering mortality events. A comparative ultrastructural study of injured and healthy I. fasciculata specimens showed that cyanobacteria disappeared from injured specimens, which suggests that cyanobacterial decay could be involved in I. fasciculata mortality. A laboratory experiment confirmed that the cyanobacteria harbored by I. fasciculata displayed a significant reduction in photosynthetic efficiency in the highest temperature treatment. The sponge disease reported here led to a severe decrease in the abundance of the surveyed populations. It represents one of the most dramatic mass mortality events to date in the Mediterranean Sea

Cyanobacteria from the Baltic Sea and Finnish lakes as an energy source and modulators of bioenergetic pathways

Cyanobacteria are a diverse group of oxygenic photosynthetic bacteria that inhabit in a wide range of environments. They are versatile and multifaceted organisms with great possibilities for different biotechnological applications. For example, cyanobacteria produce molecular hydrogen (H2), which is one of the most important alternatives for clean and sustainable energy. Apart from being beneficial, cyanobacteria also possess harmful characteristics and may become a source of threat to human health and other living organisms, as they are able to form surface blooms that are producing a variety of toxic or bioactive compounds. The University of Helsinki Culture Collection (UHCC) maintains around 1,000 cyanobacterial strains representing a large number of genera and species isolated from the Baltic Sea and Finnish lakes. The culture collection covers different life forms such as unicellular and filamentous, N2-fixing and non-N2-fixing strains, and planktonic and benthic cyanobacteria. In this thesis, the UHCC has been screened to identify potential strains for sustainable biohydrogen production and also for strains that produce compounds modifying the bioenergetic pathways of other cyanobacteria or terrestrial plants. Among the 400 cyanobacterial strains screened so far, ten were identified as high H2-producing strains. The enzyme systems involved in H2 metabolism of cyanobacteria were analyzed using the Southern hybridization approach. This revealed the presence of the enzyme nitrogenase in all strains tested, while none of them are likely to have contained alternative nitrogenases. All the strains tested, except for two Calothrix strains, XSPORK 36C and XSPORK 11A, were suggested to contain both uptake and bidirectional hydrogenases. Moreover, 55 methanol extracts of various cyanobacterial strains were screened to identify potent bioactive compounds affecting the photosynthetic apparatus of the model cyanobacterium, Synechocystis PCC 6803. The extract from Nostoc XPORK 14A was the only one that modified the photosynthetic machinery and dark respiration. The compound responsible for this effect was identified, purified, and named M22. M22 demonstrated a dual-action mechanism: production of reactive oxygen species (ROS) under illumination and an unknown mechanism that also prevailed in the dark. During summer, the Baltic Sea is occupied by toxic blooms of Nodularia spumigena (hereafter referred to as N. spumigena), which produces a hepatotoxin called nodularin. Long-term exposure of the terrestrial plant spinach to nodularin was studied. Such treatment resulted in inhibition of growth and chlorosis of the leaves. Moreover, the activity and amount of mitochondrial electron transfer complexes increased in the leaves exposed to nodularin-containing extract, indicating upregulation of respiratory reactions, whereas no marked changes were detected in the structure or function of the photosynthetic machinery. Nodularin-exposed plants suffered from oxidative stress, evidenced by oxidative modifications of various proteins. Plants initiated strategies to combat the stress by increasing the levels of alpha-tocopherol, mitochondrial alternative oxidase (AOX), and mitochondrial ascorbate peroxidase (mAPX).

Biodegradation of glyphosate in rhizospheric soil cultivated with Glycine max, Canavalia ensiformis e Stizolobium aterrimum

Biodegradation of glyphosate was evaluated in rhizospheric soil cultivated with Glycine max (soybean, var. BRS245-RR), Canavalia ensiformis and Stizolobium aterrimum. After these species were cultivated for 60 days, soil samples were collected, placed in flasks and treated with 14C-glyphosate. After 30 days of incubation, the total release rate of C-CO2 was determined along with microbial biomass (MBC), metabolic quotient (qCO2), and degradation percentage of the radio-labeled glyphosate released as 14C-CO2. A higher mass of rhizosphere-associated microorganisms was verified in the soil samples from pots cultivated with soybean, regardless of glyphosate addition. However, in the presence of the herbicide, this characteristic was the most negatively affected. Microorganisms from the C. ensiformis rhizosphere released a lower amount of 14C-CO2, while for those originated from S. aterrimum, the amount released reached 1.3% more than the total carbon derived from the respiratory activity. The rhizospheric soil from S. aterrimum also presented higher glyphosate degradation efficiency per microbial biomass unit. However, considering qCO2, the microbiota of the rhizospheric soil cultivated with soybean was more efficient in herbicide degradation.

Algas planctônicas do reservatório da Pampulha (MG): Euglenophyta, Chrysophyta, Pyrrophyta, Cyanobacteria

Neste trabalho foram descritos 28 táxons de algas planctônicas pertencentes às divisões Euglenophyta, Chrysophyta, Pyrrophyta e Cyanobacteria, encontradas no reservatório da Pampulha, estado de Minas Gerais. Foram analisadas amostras no período de maio/1992 a fevereiro/1997. Em número de espécies, Euglenophyta foi a que apresentou maior número (13 espécies em uma família), seguidas de Cyanobacteria (oito espécies), Pyrrophyta (cinco espécies) e Chrysophyta (duas espécies). Vinte e três táxons são primeiras citações para o estado de Minas Gerais.

Cylindrospermopsis raciborskii (Woloszynska) Seenayya & Subba Raju(Cyanobacteria): variação semanal e relações com fatores ambientais em um reservatório eutrófico, São Paulo, SP, Brasil

Florações de Cylindrospermopsis raciborskii têm sido cada vez mais freqüentes em reservatórios brasileiros em virtude da sua alta competitividade em ambientes tropicais eutrofizados. Esta espécie é produtora de toxinas extremamente agressivas, as quais podem causar problemas de saúde pública e danos ao meio ambiente. Nosso objetivo foi acompanhar a variação temporal da densidade desta população relacionando-a com variáveis ambientais e detectando os possíveis fatores que interferem no seu desenvolvimento. O lago estudado localiza-se na zona sul do Município de São Paulo (23º39' S e 46º37' W). É um corpo d'água eutrofizado e vem apresentando florações freqüentes de C. raciborskii. Amostras de água foram coletadas semanalmente, na parte mais profunda do lago, durante o período de setembro/97 a setembro/98. Foram analisados o perfil térmico, zona de mistura, transparência, zona eufótica, turbidez, pH, alcalinidade, condutividade, oxigênio dissolvido, nutrientes, clorofila a, densidade da população e densidade total da comunidade fitoplanctônica. A ordenação dos dados abióticos e bióticos foi feita através da análise de componentes principais (PCA) e análise de correspondência canônica (CCA). Durante os meses de novembro e dezembro/97 foram registrados os mais altos valores de densidade de C. raciborskii. Entre 10/12 e 17/12, a espécie contribuiu com 50% para a densidade total da comunidade. Nesse período foram registradas estratificações térmicas, elevados valores de temperatura da água e de turbidez e baixos valores de transparência da água. Os meses de outono e inverno de 1998 foram caracterizados pela ausência da espécie estudada, desestratificação térmica da coluna d'água, baixas temperaturas da água, altos valores de transparência (os maiores já registrados para o ambiente) e pelo aumento considerável dos bancos da macrófita aquática flutuante Eichhornia crassipes (Mart.) Solms. Tais interrelações são discutidas no presente trabalho.

Planktic Cyanobacteria from São Paulo State, Brazil: Chroococcales

The biodiversity studies of planktic cyanobacteria started in 1997 with intensive collecting in different water bodies of São Paulo State. Due to the problems brougth by eutrophication and cyanobacterial blooms, the samplings were concentrated in reservoirs of Alto Tietê region that supply drinking water to millions of people. The samples were collected with 20 µm plankton net or Van Dorn's bottle. Part of each one was preserved in formaldehyde or lugol solution and part was isolated. The culture strains were kept in BG11 and/or AMS1 media. Based on natural and culture material, 26 species were identified belonging to the families Chroococcaceae (2 taxa), Merismopediaceae (12), Microcystaceae (7) and Synechococcaceae (5). Among these species, six are potentialy toxic: Aphanocapsa incerta (Lemmerm.) Cronberg & Komárek, Microcystis aeruginosa (Kütz.) Kütz., M. botrys Teiling, M. panniformis Komárek et al., M. wesenbergii (Komárek) Komárek, and Radiocystis fernandoi Komárek & Komár.-Legn. Bacularia and Coelosphaeriopsis are genera reported for the first time in Brazil.

Effect of nitrate and ammonium on the growth and protein concentration of Microcystis viridis Lemmermann (Cyanobacteria)

Cyanobacteria are a very important group in aquatic systems, particularly in eutrophic waters. Therefore studies about their success in the environment are essential. Many hypotheses have tried to explain the dominance of Cyanobacteria, and several emphasized the importance of various nitrogen sources for the success of the group. In this study, we measured the effect of ammonium and nitrate on the growth and protein concentration of Microcystis viridis (Cyanobacteria). This species is well-known because bloom formation in eutrophic waters. The study was carried out, in experimental batch cultures, using the WC medium with different nitrogen sources: ammonium, nitrate, ammonium + nitrate (50% ammonium + 50% nitrate) and ammonium at different concentrations (to test for possible NH4+ toxicity). Protein, ammonium and nitrate concentrations were measured at end of each experiment, whereas samples for cell counts were taken daily. Results showed that Microcystis viridis grew faster with ammonium (µ = 0.393 day-1) than with nitrate (µ = 0.263 day-1) and ammonium + nitrate (µ = 0.325 day-1). This pattern is explained by the metabolism of ammonium that presents higher uptake and assimilation rates than nitrate. Maximum cell concentration, however, was higher in the ammonium + nitrate treatment, followed by nitrate treatment. Higher protein concentration were observed in the treatment with nitrate. In the ammonium toxicity test, no difference between the control and NH4+ at 50% was found. Thus, the ammonium concentrations used in these experiments were not toxic. Our results suggest that Cyanobacteria is able to grow on both nitrogen sources even if ammonium may allow faster growth and bloom development.