Aerobic granulation of pure bacterial strain

The objective of this study is to cultivate aerobic granules by pure bacterial strain, Bacillus thuringiensis, in a sequencing batch reactor. Stable granules sized 2.0–2.2 mm were formed in the reactor after a five-week cultivation. These granules exhibited excellent settling attributes, and degraded phenol at rates of 1.49 and 1.19 g phenol/(g VSS·d) at 250 and 1500 mg/L of phenol concentration, respectively. Confocal laser scanning microscopic test results show that Bacillus thuringiensis was distributed over the initial small aggregates, and the outer edge of the granule was away from the core regime in the following stage.     

Assessment of the in vivo genotoxicity of a new formulation of Bacillus thuringiensis var israelensis SH-14

Biological control agents have become a useful alternative for the reduction of the use of chemical insecticides. LABIOFAM (Cuba) is developing a new formulation of a biolarvicide that possesses as active biological agent Bacillus thuringiensis var israelensis serotype H14. In order to evaluate the genotoxicity of this new formulation, an in vivo battery test was used: micronucleus (MN), chromosome aberrations (CAs), and sperm morphology (SM) assays. A dose of 6.45?×?10⁸ spores was administered per animal via oral administration. Bone marrow cells were collected 24?h after a two day treatment for the MN assay, and 24?h after a unique treatment for the CA assay, using cyclophosphamide as the positive control. Sperm cells were collected at 5 weeks from the first of five administrations for the SM test, using acrylamide as positive control. Bacillus thuringiensis var israelensis serotype H14 failed to show either a significative increase of micronucleated polychromatic erythrocytes, chromosomal aberrations, or sperm abnormalities. Acute oral administration of a high dose of Bacillus thuringiensis var israelensis serotype H14 did not produce mutagenic effects in bone marrow or sperm cells.     

Influence of environmental factors on biodegradation of quinalphos by <Emphasis Type="Italic">Bacillus thuringiensis</Emphasis>

Background

The extensive and intensive uses of organophosphorus insecticide—quinalphos in agriculture, pose a health hazard to animals, humans, and environment because of its persistence in the soil and crops. However, there is no much information available on the biodegradation of quinalphos by the soil micro-organisms, which play a significant role in detoxifying pesticides in the environment; so research is initiated in biodegradation of quinalphos.

Results

A soil bacterium strain, capable of utilizing quinalphos as its sole source of carbon and energy, was isolated from soil via the enrichment method on minimal salts medium (MSM). On the basis of morphological, biochemical and 16S rRNA gene sequence analysis, the bacterium was identified as to be Bacillus thuringiensis. Bacillus thuringiensis grew on quinalphos with a generation time of 28.38 min or 0.473 h in logarithmic phase. Maximum degradation of quinalphos was observed with an inoculum of 1.0 OD, an optimum pH (6.5–7.5), and an optimum temperature of 35–37 °C. Among the additional carbon and nitrogen sources, the carbon source—sodium acetate and nitrogen source—a yeast extract marginally improved the rate of degradation of quinalphos.

Conclusions

Display of degradation of quinalphos by B. thuringiensis in liquid culture in the present study indicates the potential of the culture for decontamination of quinalphos in polluted environment sites.

     

Bacillus thuringiensis pesticide exposures reported to Texas poison centers

One of the most common types of microbial pesticides used is Bacillus thuringiensis. Although generally considered safe, exposures to B. thuringiensis pesticide do occur where management by healthcare providers is sought. The objective of this investigation was to describe B. thuringiensis pesticide exposures reported to Texas poison centers. Cases were all B. thuringiensis pesticide exposures reported to Texas poison centers during 2000–2010. The distribution of cases was determined for various demographic and clinical factors. Of 115 total cases, 64% occurred during May–July. The rate per 1,000,000 population was 14.3 in central Texas and 4.4 in the rest of the state. Of the patients, 56% were of an age 20 years or older and 55% were male. The exposure routes were 57% by ingestion, 30% by dermal contact, and 11% by inhalation. The patient was managed on site in 93% of the exposures and only one exposure was judged to be potentially serious. B. thuringiensis pesticide exposures reported to Texas poison centers were more frequently reported from the central region of the state. The majority of exposures occurred during April–July, involved adults and males, occurred by ingestion or dermal route, were managed on site and did not result in serious outcomes.     

Nitrifying and denitrifying bacteria in aerobic granules formed in sequencing batch airlift reactors

The purpose of this study was to investigate nitrifying bacteria and denitrifying bacteria isolated from aerobic granules. Aerobic granules were formed in an internal-circulate sequencing batch airlift reactor (SBAR) and biodegradation of NH₃ ^?-N was analyzed in the reactor. Bacteria were isolated and determined from aerobic granules using selected media. The growth properties and morphology of bacteria colonies were observed by controlling aerobic or anaerobic conditions in the culture medium. It was found that bacteria in aerobic granules were diverse and some of them were facultative aerobes. The diversity of bacteria in aerobic granules was a premise of simultaneous nitrification and denitrification.     

Effect of dilution rate on dynamic and steady-state biofilm characteristics during phenol biodegradation by immobilized <Emphasis Type="Italic">Pseudomonas desmolyticum</Emphasis> cells in a pulsed plate bioreactor

Pulsed plate bioreactor (PPBR) is a biofilm reactor which has been proven to be very efficient in phenol biodegradation. The present paper reports the studies on the effect of dilution rate on the physical, chemical and morphological characteristics of biofilms formed by the cells of Pseudomonas desmolyticum on granular activated carbon (GAC) in PPBR during biodegradation of phenol. The percentage degradation of phenol decreased from 99% to 73% with an increase in dilution rate from 0.33 h^–1 to 0.99 h^–1 showing that residence time in the reactor governs the phenol removal efficiency rather than the external mass transfer limitations. Lower dilution rates favor higher production of biomass, extracellular polymeric substances (EPS) as well as the protein, carbohydrate and humic substances content of EPS. Increase in dilution rate leads to decrease in biofilm thickness, biofilm dry density, and attached dry biomass, transforming the biofilm from dense, smooth compact structure to a rough and patchy structure. Thus, the performance of PPBR in terms of dynamic and steady-state biofilm characteristics associated with phenol biodegradation is a strong function of dilution rate. Operation of PPBR at lower dilution rates is recommended for continuous biological treatment of wastewaters for phenol removal.

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The effect of low concentrations of phenol and ammonia on egg production rates,fecal pellet production and egg viability of the calanoid copepod Acartia clausi

Estimates of daily fecundity, hatching success and fecal pellet production are reported for Acartia clausi females exposed for 10 d to low levels of phenol and ammonia. Copepods were collected in 1991 and 1992 from a southern coastal area of the Mar Grande of Taranto (southern Italy). A reduction in egg numbers and fecal pellet production was observed for females after 8 d of exposure to 500 g l^-1 phenol concentration. Ammonia (120 g l^-1) produced a significant increment in egg production, but hatching success was reduced by about 50% after nine exposure days. A. clausi was more sensitive to ammonia than phenol at high concentrations (24-h LC₅₀ phenol-32.26 mg l^-1; 24-h LC₅₀ ammonia=0.91 mg l^-1). At low concentrations, only long-term exposure to phenol determined a reduction in fecundity.     

Early murine immune responses from endotracheal exposures to biotechnology-related Bacillus strains

An immunology-based in vivo screening regime was used to assess the potential pathogenicity of biotechnology-related microbes. Strains of Bacillus cereus (Bc), Bacillus subtilis (Bs), Bacillus thuringiensis (Bt), and Bt commercial products (CPs) were tested. Balb/c mice were endotracheally instilled with purified spores, diluted CP, or vegetative cells (VC) (live or dead). Exposed mice were evaluated for changes in behavioral and physical symptoms, bacterial clearance, pulmonary granulocytes, and pulmonary and circulatory pyrogenic cytokines (interleukins (IL)-1β, IL-6 and tumor necrosis factor (TNF)-α), as well as acute phase biomarkers (fibrinogen and serum amyloid A). Except for some differences in clearance rates, no marked effects were observed in mice exposed to any spore at 10⁶ or 10⁷ colony forming units (cfu). In contrast, live Bc or Bt VCs (10⁵ or 10⁶?cfu) produced shock-like symptoms (lethargy, hunched appearance, ruffled fur, and respiratory distress), and 11–200-fold elevations in pyrogenic cytokines at 2-h post-exposure. In the study, 4-h effects included increased lethargy, ocular discharge, and 1.5–4-fold rise in circulatory acute phase markers, but no indications of recovery. Bs VC did not produce any changes in symptoms or biomarkers. After 2 or 4?h of exposure to dead VC, increases of only plasma IL-1β and TNF-α (4.6- and 12.4-fold, respectively) were observed. These findings demonstrate that purified spores produced no marked effects in mice compared to that of metabolically active bacteria. This early screening regime was successful in distinguishing the pathogenicity of the different Bacillus species, and might be useful for assessing the relative hazard potential of other biotechnology-related candidate strains.     

Bioleaching of spent Ni-Cd batteries and phylogenetic analysis of an acidophilic strain in acidified sludge

Biohydrometallurgy is a novel method to recycle discarded batteries, in which sewage sludge is used as microorganisms and culture due to the presence of indigenous Thiobacilli. A two-step continuous flow leaching system consisting of an acidifying reactor and a leaching reactor was introduced to achieve the bioleaching of spent nickel-cadmium (Ni-Cd) batteries. The acid supernatant produced in the acidifying reactor by the microorganisms with ferrous ions as the substrate was conducted into the leaching reactor to dissolve electrode materials. The efficiency of a batch treatment of batteries was examined. The results showed that the complete dissolution of two AA-sized Ni-Cd batteries with 0.6 L/d acid supernatant took about 30, 20, and 35 days for Ni, Cd, and Co, respectively. But the dissolution ability of the three metals was different. Cd and Co can be leached mostly for pH below 4.0 while the complete dissolution of Ni can be achieved for pH below 2.5. Meanwhile, a strain (named Thiooxidans. WL) accounting for the reduction of pH in the acidified sludge was isolated and sequenced. It was identified to be 100% similar to Acidithiobacillus ferrooxidans strain Tf-49 based on 16S rDNA sequence analysis. The relevant phylogenetic tree constructed indicates that the strain should be classified into genus Acidithiobacillus ferrooxidans.     

Roles of acid-producing bacteria in anaerobic digestion of waste activated sludge

Three acid-producing strains, AFB-1, AFB-2 and AFB-3, were isolated during this study, and their roles in anaerobic digestion of waste activated sludge (WAS) were evaluated. Data of 16S rRNA method showed that AFB-1 and AFB-2 were Bacillus coagulans, and AFB-3 was Escherichia coli. The removal in terms of volatile solids (VS) and total chemical oxygen demand (TCOD) was maximized at 42.7% and 44.7% by inoculating Bacillus coagulans AFB-1. Besides, the optimal inoculum concentration of Bacillus coagulans AFB-1 was 30% (v/v). Solubilization degree experiments indicated that solubilization ratios (SR) of WAS reached 20.8%±2.2%, 17.7%±1.48%, and 11.1%±1.53%. Volatile fatty acids (VFAs) concentrations and compositions were also explored with a gas chromatograph. The results showed that VFAs improved by 98.5%, 53.0% and 11.6% than those of the control, respectively. Biochemical methane potential (BMP) experiments revealed that biogas production increased by 90.7% and 75.3% when inoculating with Bacillus coagulans AFB-1 and AFB-2. These results confirmed that the isolated acid-producing bacteria, especially Bacillus coagulans, was a good candidate for anaerobic digestion of WAS.

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Model of a chemostat utilising phenol as inhibitory substrate

A pure culture of Acinetobacter lwoffi was grown in a batch and continuous culture using phenol as the limiting substrate. Five kinetic models were applied to the data to determine the kinetic parameters governing growth of the organisms. A non-linear leastsquares technique was used to fit the data to the different functions. No significant differences were found on statistical basis between the models and so a choice was made on the grounds of mathematical simplicity. The so-called ‘Haldane function’ was chosen as the best and utilised in the model describing the steady-state behaviour of the chemostat. This model takes into account the inhibitory character of phenol and also the maintenance energy of the bacteria. Analysis of local stability shows that one can expect to obtain three steady states of which one is the trivial washout state, another is unstable and the third is a high conversion stable steady state.This model was tested with a pure culture of Acinetobacter lwoffi and phenol inlet concentrations of 200, 500 and 1000 mgl^?1. Good agreement was found between the model and the results of the continuous experiments.     

Study of Fishes in the Chung-Kung Stream: Species Distribution and Organochlorine Pesticide Residues

Abstract

In this study, we investigated the potential for reductive dechlorination of 2,4-dichlorophenol (2,4-DCP), 2,4,6-trichlorophenol (2,4,6-TCP), and pentachlorophenol (PCP) by municipal sewage sludge adapted to 2,4-DCP at different concentrations. 2,4-DCP was completely dechlorinated within 4 weeks. After 18 weeks' incubation, 2,4,6-TCP was also completely dechlorinated and the residue of PCP was 0, 44.46, 51.96% at 0.5, 5, and 50 μg ml¹ respectively. the 2,4-DCP adapted communities initially removed the ortho-chlorine from PCP of 5.0, 50 μg ml¹, following an ortho < para < meta order of chlorine removal. Intermediate products were 3,4,5-TCP, 3,5-DCP, 3-CP (3-chlorophenol), phenol, benzoate and hexanoic acids, whereas PCP (0.5 μg ml¹) indicated a preference for meta-chlorine removal. the intermediate product of 2,4,6-TCP at three concentrations were 2,4-DCP, phenol, benzoate and hexanoic acid. These products were identified by GC-MASS spectrometry. the effects of supplements, including sodium citrate (0.08 mM), sodium pyruvate (0.18 mM), sodium sulphate (0.14 mM) had a direct stimulatory effect on the dechlorination of 2,4,6-TCP and PCP after treatment for 4 weeks, but dechlorination was inhibited after 8 weeks.     

温度,碳,氮,磷对一株芽孢杆菌生长的影响

A series of orthogonal array tests and single-factor tests were designed to study the growth of Bacillus sp. H strain isolated from Microcystis aeruginosa in the Taihu Lake. The results indicated that within the experimental scope the factors affecting the growth of Bacillus sp. H strain were arranged in the order organic carbon>temprature>phosphate>ammonium. Of the four factors, organic carbon and temperature most significantly affected the growth of Bacillus sp. H strain. It was also found that with the raising of temperature, concentration of organic carbon or the ratio of N/P, the growth of Bacillus sp. H strain would become better. The optimum conditions for the growth of Bacillus sp. H strain in this experiment weretemperature 30℃, concentration of ammonium 0.520 g/L , concentration of peptone 2.0 g/L , phosphate 0.088 g/L , and organic carbon 2.28 g/L. Fig 4, Tab 2, Ref 5     

Barnacles: Possible indicators of zinc pollution?

Barnacles [Balanus balanoides (L.), Elminius modestus Darwin, Lepas anatifera (L.)] from several different sites were found to accumulate the heavy metal zinc. The majority of the zinc was deposited in the tissues associated with the gut, and the level of zinc in soft body tissue generally reflected well the level of zinc in the immediate sea-water environment. The zinc accumulated in the gut tissues was in the form of discrete granules, mainly within the parenchyma cells which surround the gut. These granules probably exist as an insoluble zinc salt. A comparison of the zinc level in barnacles with those recorded for some other marine organisms indicates the possible usefulness of barnacles as indicators of zinc pollution.     

Absorption of atmospheric phenol by various tree species and their tolerance to phenol

To assess the effect of tree planting on atmospheric phenol, a study was made on the absorption of phenol by various tree species and the tolerance of these species to phenol. The absorption rates ranged from 21.3 (camellia) to 129 ng dm^‐2h^‐1 ppb^‐1 (Japanese elm) at 1000 μmol of photons m^‐2 s^‐1, and the absorption rate increased in the following order: coniferous tree species ? evergreen broad‐leaved tree species < deciduous broad‐leaved tree species. When the light intensity was varied, a linear relationship between the phenol absorption rate and the transpiration rate was observed for three tree species. In comparison with the absorption rate estimated from a simplified gas diffusive resistance model, we conclude that phenol is absorbed through the stomata and is metabolized fairly rapidly within the leaf tissue, although the absorption rate is less than the estimated potential absorption rate. At phenol concentrations below 200 ppb, the tree can absorb atmospheric phenol for at least 8 h without any visible foliar injury. Trees in general could act as an important sink for atmospheric phenol at phenol concentrations less than 200 ppb, a concentration about twenty times higher than normal ambient levels.     

Zinc phosphate granules in tissue surrounding the midgut of the barnacle Balanus balanoides

The chemical composition of inorganic granules found in parenchyma cells surrounding the midgut of adult Balanus balanoides (L.) was determined. X-ray microanalysis indicated the presence of magnesium, phosphorus, potassium, calcium, iron and zinc in the granules. A quantitative analysis of granule-rich pellets isolated from the midguts of adult barnacles showed that the granules were composed mainly of zinc phosphate, the remaining elements being minor constituents only.     

Isolation and characterization of granules from the kidney of the bivalveMercenaria mercenaria

Intracellular and extracellular granules are found in most bivalve kidneys. We examined the composition of kidney granules from the infaunal bivalveMercenaria mercenaria (L.) collected in 1985 from Cataumet Bay, Bourne, and Waquoit Bay, Falmouth, Massachusetts, USA. Small granules are numerically dominant, but large granules dominate the samples on the basis of weight. Large granules (estimated diam. >15 m) are composed primarily of metals (28% by weight) with Mn (8%), Ca (8%), Zn (4%) and Fe (4%) as the major contributors. Non-metal elements (P, C, H, N, S) together constituted 21% of the granule dry weight, though organic content (0.02% protein, 0.3% carbohydrate and 0.5% lipid) was low. Small kidney granules (10 m diam.) and digestive gland granules fromM. mercenaria had significantly higher C, H, and N contents than larger kidney granules. Our results, taken together with those from other bivalve species, suggest that kidney granule formation and subsequent increase in size is a continuous process of lysosomal maturation, residual body release and extracellular accumulation of predominately inorganic elements.     

苯酚的超临界水氧化试验

以自建的一套连续式超临界水氧化装置的温度，压力，溶氧量和流量等参数的可控性进行了实验，并在此基础上进行了苯酚的超临界水氧化研究。结果表明：所建立的装置能比较精确控制各种条件参数，是可靠的超临界水氧化试验研究装置。在氧气过量的条件下，停留时间是影响苯酚在超临界水中氧化分解的主要因素，反应的温度和压力升高导致苯致苯酚去除率的增大，只要有足够的氧气，苯酚起始浓度的增加对苯酚的转化率影响不明显，在实验条件下，溶液中苯酚的去除动力学对苯酚是一级，氧气是零级。     

Accumulation of copper within the “Hepatopancreatic” caeca of Corophium volutator (Crustacea: Amphipoda)

The concentrations of various metals in specimens of Corophium volutator (Pallas) which have been taken from a site that receives freshwater drainage with a high concentration of copper, zinc and manganese, are compared with those in individuals from a site with a normal concentration of these metals for coastal waters. The amphipods at the contaminated site contain 259 g g⁺¹ dry weight of copper, which is three times that of individuals from the uncontaminated site. The concentrations of the other metals do not differ substantially between the two populations. The hepatopancreatic caeca of the midgut of amphipods with a high concentration of copper have numerous granules of homogeneous electron-dense material which can aggregate within the multivesicular bodies. Some of the amphipods with a lower concentration of copper have similar granules, but these are fewer in number. X-ray microporbe analysis shows that these granules contain copper and sulphur, and occasionally calcium. Various tests on the granules in frozen section indicate that they are relatively insoluble in a wide range of reagents and that the copper is probably complexed with organic matter.     

Combined effects of copper and cadmium on Chlorella pyrenoidosa H.Chick: subcellular accumulation,distribution, and growth inhibition

Disposal of waste into aquatic ecosystems may cause microalgae to be exposed to various metals, e.g. copper and cadmium. The effects caused by combinations of metals may be more serious. Evaluations of subcellular fate, bioaccumulation, and biological effects of metals on aquatic organisms are generally derived from experiments with individual metals. The present study aims to evaluate the effects of exposure of Chlorella pyrenoidosa to copper and cadmium in combination on subcellular accumulation, distribution, and growth. The algae were exposed for 72 h to copper at concentrations of 13 – 25 µmol L^?1, cadmium at about 6 µmol L^?1, and combinations thereof. The levels of copper and cadmium in subcellular organelles, heat-denaturated protein, metal-rich granules, and heat-stable protein were determined by atomic absorption spectrometry. Exposure of C. pyrenoidosa to copper and cadmium in combination inhibited growth more strongly than copper and cadmium individually. Highest accumulation was observed in metal-rich granules and heat-stable proteins. Administration of both metals in combination affected their subcellular distribution: copper was mainly distributed into the metal-rich granules (70%–80%) and heat-stable proteins (9%–24%), cadmium in the metal-rich granules (88%–98%).