Factors influencing the inhibition of aflatoxin production in corn by Aspergillus niger

Aspergillus niger, a mold commonly associated with Aspergillus flavus in damaged corn, interferes with the production of aflatoxin when grown with A. flavus on autoclaved corn. The pH of corn-meal disks was adjusted using NaOH-HCl, citric acid-sodium citrate, or a water extract of A. niger fermented corn. Aflatoxin formation was completely inhibited below pH 2.8-3.0, irrespective of the system used for pH adjustment. When grown in association with A. flavus NRRL 6432 on autoclaved corn kernels, A. niger NRRL 6411 lowered substrate pH sufficiently to suppress aflatoxin production. The biodegradation of aflatoxin B1 or its conversion to aflatoxin B2a were eliminated as potential mechanisms by which A. niger reduces aflatoxin contamination. A water extract of corn kernels fermented with A. niger caused an additional inhibition of aflatoxin formation apart from the effects of pH.     

Enzymatic release of phosphate in water from subarctic lakes in Northern Sweden

Enzymatic phosphate release was studied in water from three small lakes in the subarctic region of northern Sweden. Free dissolved hydrolytic enzymes were shown to release considerable amounts of phosphate from autoclaved lake water. It was possible to show that phosphate was enzymatically released from particulate matter by exposure of autoclaved, ³²P-labelled plankton to lake enzymes followed by characterization of degradation products with gel chromatography. Seston turned out to be vitally important as a phosphorus source for planktic organisms since plankton inoculated in lake water freed from seston were shown to produce high activities of phosphatase.     

Abscisic Acid and Ethylene Interact in Rice Spikelets in Response to Water Stress During Meiosis

This study investigated whether and how the interaction between abscisic acid (ABA) and ethylene is involved in the regulation of rice (Oryza sativa L.) spikelet sterility when subjected to water stress during meiosis. Two rice cultivars, HA-3 (drought-resistant) and WY-7 (drought-susceptible), were used and subjected to well-watered and water-stressed (WS) treatments during meiosis (15–2 days before heading). Leaf water potentials of both cultivars markedly decreased during the day as a result of the WS treatment, but panicle water potentials remained constant. The percentage of sterile spikelets in WS plants was increased by 49.7% for WJ-7 but only 12.7% for HA-3. ABA, ethylene, and 1-aminocyclopropane-1-carboxylic acid were all enhanced in spikelets by the water stress, but ethylene was enhanced more than ABA in WY-7 when compared with that in HA-3. Spikelet sterility was significantly reduced when ABA or amino-ethoxyvinylglycine, an inhibitor of ethylene synthesis, was applied to the panicles of WS plants at the early meiosis stage. Application of ethephon, an ethylene-releasing agent, or fluridone, an inhibitor of ABA synthesis, had the opposite effect, and sterility was increased. The results suggest that antagonistic interactions between ABA and ethylene may be involved in mediating the effect of water stress on spikelet fertility. A higher ratio of ABA to ethylene would be a physiologic trait of rice adaptation to water stress.     

Inactivation of poliovirus type 1 in mixed human and swine wastes and by bacteria from swine manure.

The persistence of poliovirus type 1 (PO1) in mixed septic tank effluent and swine manure slurry was determined, and the antiviral effects of several bacterial cultures isolated from swine manure slurry were demonstrated. In two field experiments, PO1 was consistently inactivated more rapidly in the mixed waste than in the control Dulbecco's phosphate-buffered saline (D-PBS). D values (time [in days] for a 90% reduction of virus titer) were 18.7 and 29.9 for the mixed waste and 56.5 and 51.8 for the D-PBS control, respectively. The virus inactivation in the mixed waste was temperature dependent. A comparison of PO1 inactivation in raw mixed waste, autoclaved mixed waste, and bacterium-free filtrate of raw mixed waste at the same pH and temperatures provided an initial demonstration that the virus inactivation in the mixed waste is related, at least in part, to microbial activity. At 25 degrees C, the D value was 6.8 for the mixed waste, 11.2 for the autoclaved mixed waste, and 10.5 for the bacterium-free filtrate of raw mixed waste. At 37 degrees C, D values were 1.3, 3.9, and 3.1 for these three suspending media, respectively. Three bacterial isolates which had shown antiviral effects in a screening test each caused virus inactivation in autoclaved mixed waste, in which the effect of other microorganisms was excluded. Inhibition of PO1 inactivation by protease inhibitors suggests that the virus inactivation in the mixed waste was due in part to proteolytic enzymes produced by bacteria in the waste.     

A new method for the production of sterile colonies of Lucilia sericata

Maggot debridement therapy (MDT) refers to the use of blowfly larvae to clean or debride an infected wound. Most commonly, larvae of Lucilia sericata (Meigen) (Diptera: Calliphoridae) are used, and are sterilized prior to use to ensure no further bacterial infections are introduced during treatment. Current methods sterilize eggs from laboratory‐reared blowfly colonies, after which sterile early second instar maggots can be provided to hospitals for use in treatment. Maggots not required for treatment are used for colony regeneration, in which sterility is not maintained. The ability to maintain sterility beyond this would allow further research into fly–bacteria interactions and the effects of different bacteria on the blowfly lifecycle. This study aimed to produce a colony of sterile adults, using current egg sterilization practice, but maintaining sterility through to pupation and emergence. The production of a sterile colony allows further research into the impact of bacteria on fly development and survival. Eggs were placed on a sterile food source within autoclaved plant tissue culture containers to allow growth under sterile conditions. Nutrient agar plating of sterilized and non‐sterilized eggs, larvae and adults (post‐emergence), as well as the pupation medium and feed source in nutrient broth confirmed the aerobic sterility of all samples involved. The lifecycle of L. sericata was successfully completed through pupation to emergence with no effects on lifespan or oviposition by the newly emerged, sterile adult colony.     

Inactivation of poliovirus type 1 in mixed human and swine wastes and by bacteria from swine manure.

The persistence of poliovirus type 1 (PO1) in mixed septic tank effluent and swine manure slurry was determined, and the antiviral effects of several bacterial cultures isolated from swine manure slurry were demonstrated. In two field experiments, PO1 was consistently inactivated more rapidly in the mixed waste than in the control Dulbecco's phosphate-buffered saline (D-PBS). D values (time [in days] for a 90% reduction of virus titer) were 18.7 and 29.9 for the mixed waste and 56.5 and 51.8 for the D-PBS control, respectively. The virus inactivation in the mixed waste was temperature dependent. A comparison of PO1 inactivation in raw mixed waste, autoclaved mixed waste, and bacterium-free filtrate of raw mixed waste at the same pH and temperatures provided an initial demonstration that the virus inactivation in the mixed waste is related, at least in part, to microbial activity. At 25 degrees C, the D value was 6.8 for the mixed waste, 11.2 for the autoclaved mixed waste, and 10.5 for the bacterium-free filtrate of raw mixed waste. At 37 degrees C, D values were 1.3, 3.9, and 3.1 for these three suspending media, respectively. Three bacterial isolates which had shown antiviral effects in a screening test each caused virus inactivation in autoclaved mixed waste, in which the effect of other microorganisms was excluded. Inhibition of PO1 inactivation by protease inhibitors suggests that the virus inactivation in the mixed waste was due in part to proteolytic enzymes produced by bacteria in the waste.     

Microbial challenges for domestic heating oil storage tanks

Microbial growth on hydrocarbons is common in nature and used in bioremediation of contaminated sites, whereas in fuel storage tanks this phenomenon can affect the stability of the fuel and the tank. The impact of microbial growth and produced metabolites on materials, which are used in the construction of storage tanks, were analyzed. In contrast to metal tank components, polymeric materials did not affect or were influenced by microorganisms. Zinc was highly corroded by microbial growth, most likely due to the formation of organic acids that were produced during microbial growth on hydrocarbons. A contaminated water phase in a storage tank of a heating system was simulated with a self‐constructed pump test bench. Microbial growth began in the water phase of the storage tank and microbes were distributed throughout the tank system, through water‐in‐oil microemulsions. No microbial growth was observed in oil that was previously contaminated, indicating that essential nutrients had been depleted. The identification and removal of these essential nutrients from fuels could minimize or prevent microbial contamination. The results are discussed with regard to developing recommendations for the design and operation of domestic heating oil storage tanks to lower the risk of technical failure due to microbial contamination.     

Design and analysis of a solar reactor for anaerobic wastewater treatment

The aim of this research was to design a solar heated reactor system to enhance the anaerobic treatment of wastewater or biological sludge at temperatures higher than the ambient air temperature. For the proposed reactor system, the solar energy absorbed by flat plate collectors was transferred to a heat storage tank, which continuously supplied an anaerobic-filter reactor with water at a maximum temperature of 35 degrees C. The packed reactor was a metallic cylindrical tank with a peripheral twin-wall enclosure. Inside this enclosure was circulated warm water from the heat storage tank. Furthermore, a mathematical model was developed for the prediction of the temperature distribution within the reactor under steady state conditions. Preliminary results based on model simulations performed with meteorological data from various geographical regions of the world suggested that the proposed solar reactor system could be a promising and environmentally friendly approach for anaerobic treatment of wastewater and biological sludge.     

Inhibitor of Clostridium perfringens Formed by Heating Sodium Nitrite in a Chemically Defined Medium

An inhibitor of Clostridium perfringens formed when low levels of nitrite were autoclaved with a defined chemical medium. A systematic study of the medium revealed that only amino acids and mineral salts were involved in the production of this inhibitor, which was proven to be a toxic compound formed from cysteine, ferrous sulfate, and sodium nitrite. The inhibitor was compared to several known compounds. S-nitrosocysteine inhibited the test organism, but would not form in the test system in amounts large enough to explain the observed inhibition. Roussin red salt was unstable in the test system and therefore was not the inhibitor. Roussin black salt, which was also inhibitory, could form in sufficient amounts to explain the inhibition. A complex of cysteine, iron, and nitric oxide was detected in the autoclaved solution of cysteine, ferrous sulfate, and sodium nitrite; this cysteine complex did not appear to be inhibitory, however, at levels which could form in the autoclaved medium. The observed inhibition may have been due to the combined effects of sublethal concentrations of each compound.     

Cadmium sorption by bacteria and freshwater sediment

Summary Sorption of cadmium by sediment bacteria and freshwater sediment was investigated using diffusion chambers to simulate the water-sediment interface. Diffusion chambers were constructed to provide two compartments separated by a dialysis membrane. Diffusion of cadmium across the membrane was monitored after pure cultures of sediment bacteria or lake sediments were added to the sediment side of a diffusion chamber. Cellular accumulation of cadmium by cadmium-sensitive and cadmium-resistant bacteria removed between 20% and 80% of the dissolved cadmium from the simulated water column and pore water. Cellular accumulation of cadmium was greatest for cadmium-sensitive isolates that were tested. Sediment with an intact microbial community sequestered 80% of the cadmium added to sediment, whereas autoclaved sediment retained 97% of the metal that was added. Addition of glucose to cadmium-amended sediment decreased retention of cadmium by untreated and autoclaved sediments, resulting in elevated concentrations of dissolved cadmium in the simulated water column.     

Zooplankton cultivation and prawn seed-production in an artificial ecosystem

The cultivation experiments were conducted in two parts: (1) zooplankton cultivation in a heterogeneous system as fundamental research (Experiment I); (2) prawn seed-production in a homogeneous system as applied research (Experiment II). In Experiment I, several types of apparatus were designed in order to study a feedback system for cultivation of zooplankton (Brachionus plicatilis andTigriopus japonicus). A 550-1 round transparent tank was used for reserve culture, A 150-1 zigzag stream unit was made of vinyl plates, and connected with the reserve culture tank. In both cases, the water was re-circulated 20 times a day by an airlift pump. A tower-decomposer tank and two blowing cultivators were specially designed for promoting energy-flow. During a 140-day experiment, total wet weight of feed and harvest were 4817 g of yeast and 4308 g of zooplankton, respectively. The rate of food conversionwas 89.4% in terms of wet weight and 33.6% in dry weight. In Experiment II, a 2500 m³-hatchery tank with movable aerators and airlift circulation systems was contructed to maintain an artificial ecosystem for rearing of the prawnPenaeus japonicus. Feeding rate in the 2500 m³-tank was 10.8% lower than that in the 200 m³-tank, but growth rate in the former was 10.6% higher than in the latter.     

Arsenic removal from groundwater by pretreated waste tea fungal biomass

Arsenic contamination in ground water poses a serious threat on human health. The tea fungus, a waste produced during black tea fermentation has been examined for its capacity to sequester the metal ions from ground water samples. Autoclaved tea fungal mat and autoclaving followed by FeCl3 pretreated tea fungal mat were exploited for removal of As(III), As(V) and Fe(II) from ground water sample collected from Kolkata, West Bengal, India. The biosorption rate tends to increase with the increase in contact time and adsorbent dosage. FeCl3 pretreated and autoclaved fungal mats removed 100% of As(III) and Fe(II) after 30 min contact time and 77% of As(V) after 90 min contact time. The optimum adsorbent dosage was 1.0 g/50 mL of water sample. The results revealed that the FeCl3 pretreated fungal mat could be used as an effective biosorbent for As(III) and As(V); autoclaved fungal mat for Fe(II) removal from ground water sample.     

Heterologous expression of the mutated melon ethylene receptor gene Cm-ERS1/H70A produces stable sterility in transgenic lettuce (Lactuca sativa)

The mutated melon ethylene receptor gene Cm-ERS1/H70A was introduced into tobacco and induced stable sterility in transgenic lines. This gene contains a missense mutation that converts the His(70) residue to Ala in the melon ethylene receptor gene Cm-ERS1. To test the applicability of this inducible sterility system to other plants, lettuce (Lactuca sativa) was transformed with the gene using Agrobacterium, and putative transformants containing Cm-ERS1/H70A were obtained. Thirteen randomly selected putative transformants were grown in a growth room under constant conditions, and seven of the lines showed sterility or significantly reduced fertility. DNA gel blot analysis confirmed the integration of the Cm-ERS1/H70A gene into the genomes of the putative transformants, and RT-PCR and protein gel blot analysis confirmed the expression of Cm-ERS1/H70A mRNA and protein in all of the transformants. Five transformants showing sterility or reduced fertility when grown in a growth room under constant conditions were randomly selected to be grown in an open-air greenhouse under various environmental conditions. All five showed stable sterility under the various conditions. These results suggest that Cm-ERS1/H70A can induce sterility in heterologous transgenic plants.     

An innovative membrane bioreactor (MBR) system for simultaneous nitrogen and phosphorus removal

Membrane filtration was integrated with a post-denitrification process to form an innovative membrane bioreactor (MBR) system for effective organic degradation and nutrient (N and P) removal. The system comprised of an aerobic tank, an anoxic tank, an intermediate sedimentation tank, and a membrane filtration tank. The sedimentation tank functioned not only as a rough settler for sludge–water separation before membrane filtration but also as an anaerobic chamber for P release. While half of the influent flowed into the aerobic tank, the other half was fed into the anoxic tank to favor the proliferation of phosphorus accumulating organisms (PAOs). The experiment was conducted continuously for about 430 days. With a short overall treatment time of less than 10 h for municipal wastewater, the MBR-based process could achieve the total organic carbon, total nitrogen, and total phosphorus removals of around 94%, 85%, and 87%, respectively. The growth and activity of PAOs in the MBR system were evidenced by the significant P release in the anaerobic chamber followed by the luxury P uptake in the membrane tank. With the DAPI and PAO_mix probe staining, the increases of PAOs and polyhydroxybutyrate (PHB) in sludge during the experiment were well observed under the fluorescent microscope.     

Simple field method for concentration of viruses from large volumes of water

A pressure spray tank was adapted to supply positive pressure for processing water samples for concentrating viruses with microporous filters under field conditions. This low-cost system allows water to be processed in locations where electric current is not readily available or where light-weight portable equipment is required.     

Simple field method for concentration of viruses from large volumes of water.

A pressure spray tank was adapted to supply positive pressure for processing water samples for concentrating viruses with microporous filters under field conditions. This low-cost system allows water to be processed in locations where electric current is not readily available or where light-weight portable equipment is required.     

Optimizing methods to estimate zooplankton concentration based on generalized patterns of patchiness inside ballast tanks and ballast water discharges

Zooplankton populations are spatially heterogeneous in nature and inside ship ballast tanks. Sampling methods should take heterogeneity into account, particularly when estimating quantitative variables such as abundance or concentration. It is particularly important to generate unbiased estimates of zooplankton concentration in ballast water when assessing compliance with new international ballast water discharge standards. We measured spatial heterogeneity of zooplankton within ballast water using three sampling methodologies. In‐tank pump samples were collected at fixed depths within the vertical part of the ballast tank (side tank). Vertical net‐haul samples were collected from the upper portion of the tank as a depth‐integrated and historically relevant method. In‐line, time‐integrated samples were collected during ballast discharge by an isokinetic sample probe, likely representing the double bottom part of the ballast tank. The bias and precision associated with each sampling method were evaluated in reference to the estimated average abundance of the entire ballast tank, which was modeled from the data collected by all methods. In‐tank pump samples provided robust evidence for vertical stratification of zooplankton concentration in the side tank. A consistent trend was also observed for in‐line discharge samples, with zooplankton concentration decreasing through time as the ballast tank is being discharged. Sample representativeness, as compared to the tank average, varied depending on the depth or tank volume discharged. In‐line discharge samples provided the least biased and most precise estimate of average tank abundance (having lowest mean squared error) when collected during the time frame of 20%–60% of the tank volume being discharged. Results were consistent across five trips despite differences in ballast water source, season, and age.     

Effect of temperature and active biogas process on passive separation of digested manure

The objective of the study was to identify the optimum time interval for effluent removal after temporarily stopping stirring in otherwise continuously stirred tank reactors. Influence of temperature (10 and 55 degrees C) and active biogas process on passive separation of digested manure, where no outside mechanical or chemical action was used, within the reactor was studied in three vertical settling columns (100 cm deep). Variations in solids and microbial distribution at top, middle and bottom layers of column were assessed over a 15 day settling period. Results showed that best solids separation was achieved when digested manure was allowed to settle at 55 degrees C with active biogas process (pre-incubated at 55 degrees C) compared to separation at 55 degrees C without active biogas process (autoclaved at 120 degrees C, for 20 min) or at 10 degrees C with active biogas process. Maximum solids separation was noticed 24h after settling in column incubated at 55 degrees C, with active biogas process. Microbiological analyses revealed that proportion of Archaea and Bacteria, absent in the autoclaved material, varied with incubation temperature, time and sampling depth. Short rod shaped bacteria dominated at 55 degrees C, while long rod shaped bacteria dominated at 10 degrees C. Methanosarcinaceae were seen more abundant in the surface layer at 55 degrees C while it was seen more common in the top and bottom layers at 10 degrees C. Thus, passive separation of digester contents within the reactor can be used effectively as an operating strategy to optimize biogas production by increasing the solids and biomass retention times. A minimum of 1-2h "non-stirring" period appears to be optimal time before effluent removal in plants where extraction is batch-wise 2-4 times a day.     

Spikelet Sterility and Flowering Response of Rice to Water Stress at Anthesis

Water deficits at the anthesis stage of rice (Oryza sativa L.)induce a high percentage of spikelet sterility and reduce grainyield. This study attempted to elucidate the direct effectsof water stress on panicle exsertion, spikelet opening, andspikelet desiccation leading to spikelet sterility. A well-wateredtreatment and two water stress levels were imposed in pot-grownplants of IRAT 13 (upland cultivar) and IR20 (lowland cultivar)at the time of flowering under greenhouse conditions A cultivar difference was observed in the flowering responseto water stress with a high sensitivity in IR20. The time courseof panicle exsertion showed an inhibitory effect due to thelow panicle water status. Low panicle water potentials significantlyreduced the number of opened spikelets. Spikelet opening wascompletely inhibited at panicle water potentials below –1·8MPa and –2·3 MPa in IR20 and IRAT 13, respectively.However, the peak spikelet opening time in a day was not influencedby the stress treatment. Spikelets in stressed panicles wereobserved to remain open for a longer period than in the well-wateredpanicles. The role of turgor in spikelet opening is also discussedin the study. At low panicle water potentials, severe desiccationof spikelets and anthers was noted. The deleterious effectsof water deficits on spikelet opening and spikelet water losscontributed to reduced spikelet fertility Oryza sativa L., rice, spikelet sterility, flowering, water stress, panicle water potentials, turgor potentials, desiccation