Microbial population of feedlot waste and associated sites

A quantitative determination was made every 2 months for a year of the microflora of beef cattle waste and runoff at a medium-sized midwestern feedlot. Counts were obtained for selected groups of organisms in waste taken from paved areas of pens cleaned daily and, therefore, reflect the flora of raw waste. Overall, in terms of viable count per gram dry weight, the feedlot waste contained 10¹⁰ total organisms, 10⁹ anaerobes, 10⁸ gram-negative bacteria, 10⁷ coliforms, 10⁶ sporeformers, and 10⁵ yeasts, fungi, and streptomycetes. The specific numbers and pattern of these groups of organisms varied only slightly during the study in spite of a wide variation in weather. Data indicate that little microbial growth occurs in the waste as it exists in the feedlot. Runoff from the pens contained the same general population pattern but with greater variation attributable to volume of liquid. Comparable determinations of an associated field disposal area (before and after cropping), stockpiled waste, and elevated dirt areas in the pens indicate that fungi, and especially streptomycetes, are the aerobic organisms most associated with final stabilization of the waste. Yeasts, which are the dominant type of organism in the ensiled corn fed the cattle, do not occur in large numbers in the animal waste. Large ditches receiving runoff and subsurface water from the fields have a population similar to the runoff but with fewer coliforms.     

Integrated System of Phytodepuration for Agroindustrial Wastewater: Three Different Case Studies

The effluents deriving from agricultural industries are sources of wastewater sensibly different from common civil wastewater treatment plants effluents, because they are characterized by significant amounts of nutrients and organic load. Agricultural industries require considerable water volumes for processing the farm products, in doing so generating huge volumes of wastewater, with high concentration of chemical oxygen demand (COD), nitrogen (N) and phosphorus (P). Advanced and low cost techniques for water depuration are required in such circumstances, as the use of Integrated System of Phytodepuration (ISP). In the present work, three different case studies (a dairy, a pig feedlot and a vinegar industry) are investigated: the performances of the ISPs were evaluated analyzing raw wastewaters and final effluents over a period ranging from 2 to 4 years. The results obtained show that the designed ISPs are characterized by a mean efficiency value higher than 85% for COD removal, 73% for N and 85% for P. Moreover, for the pig feedlot the ISP final effluent is characterized by a quality level not only suited for the release into surface waters but also for irrigation, while for the other two case studies is possible to release the final effluent in surface water.     

The investigation of effect of organic carbon sources addition in anaerobic–aerobic (low dissolved oxygen) sequencing batch reactor for nutrients removal from wastewaters

The effect of addition of organic carbon sources (acetic acid and waste activated sludge alkaline fermentation liquid) on anaerobic–aerobic (low dissolved oxygen, 0.15–0.45 mg/L) biological municipal wastewater treatment was investigated. The results showed that carbon source addition affected not only the transformations of polyhydroxyalkanoates (PHA), glycogen, nitrogen and phosphorus, but the net removal of nitrogen and phosphorus. The removal efficiencies of TN and TP were, respectively, 61% and 61% without organic carbon source addition, 81% and 95% with acetic acid addition, and 83% and 97% with waste activated sludge alkaline fermentation liquid addition. It seems that the alkaline fermentation liquid of waste biosolids generated in biological wastewater treatment plant can be used to replace acetic acid as an additional carbon source to improve the anaerobic–aerobic (low dissolved oxygen) municipal wastewater nutrients removal although its use was observed to cause a slight increase of effluent BOD and COD concentrations.     

Observations on the distinction between oligotrophic and eutrophic marine bacteria

The nutritional requirements of two marine bacteria designated as oligotrophic because they could grow on media containing 10 mg of C per liter supplied as peptone and two classified as eutrophic because they could grow only at higher concentrations of C supplied as peptone were examined. Each of the four organisms was found to have its own unique group of compounds which could serve either individually or in combination as sources of carbon and energy for growth. When the peptone in the medium was replaced by another appropriate source of carbon and energy, the difference in the capacity of the organisms examined to grow at 10 mg of C per liter disappeared, and all four organisms could be described as being oligotrophic. Some of the organisms required a low concentration of one specific carbon source but a higher concentration of another. One of the organisms was inhibited by high concentrations of one specific carbon source but not by another. The observations indicate that current methods of enumeration based on the capacity of cells to grow in the presence of high or low concentrations of complex mixtures of nutrients such as peptone do not distinguish between two broad classes of bacteria differing intrinsically in their ability to grow at high and low concentrations of nutrients. Whether two such broad classes exist seems extremely doubtful. Which organisms will multiply in a particular environment will depend on both the specific nutrients available and their concentrations.     

Cellulase Production by Trichoderma viride on Feedlot Waste

Feedlot waste contains essentially all the necessary nutrients for batch fermentation with the fungus Trichoderma viride. The organism utilizes two-thirds of the carbohydrate in feedlot waste while elaborating cellulase in quantities comparable to commercial preparations. Essentially odor-free, the fermented waste contains all of the original nitrogen but has 24% less organic matter.     

Antifungal potential of extracellular metabolites produced by Streptomyces hygroscopicus against phytopathogenic fungi

Indigenous actinomycetes isolated from rhizosphere soils were assessed for in vitro antagonism against Colletotrichum gloeosporioides and Sclerotium rolfsii. A potent antagonist against both plant pathogenic fungi, designated SRA14, was selected and identified as Streptomyces hygroscopicus. The strain SRA14 highly produced extracellular chitinase and β-1,3-glucanase during the exponential and late exponential phases, respectively. Culture filtrates collected from the exponential and stationary phases inhibited the growth of both the fungi tested, indicating that growth suppression was due to extracellular antifungal metabolites present in culture filtrates. The percentage of growth inhibition by the stationary culture filtrate was significantly higher than that of exponential culture filtrate. Morphological changes such as hyphal swelling and abnormal shapes were observed in fungi grown on potato dextrose agar that contained the culture filtrates. However, the antifungal activity of exponential culture filtrates against both the experimental fungi was significantly reduced after boiling or treatment with proteinase K. There was no significant decrease in the percentage of fungal growth inhibition by the stationary culture filtrate that was treated as above. These data indicated that the antifungal potential of the exponential culture filtrate was mainly due to the presence of extracellular chitinase enzyme, whereas the antifungal activity of the stationary culture filtrate involved the action of unknown thermostable antifungal compound(s).     

Factors influencing the production of cellulases by Sporotrichum thermophile.

Cellulase production and growth of a strain of Sporotrichum thermophile were studied by using a mineral salts medium supplemented with yeast extract and insoluble cellulose. The effects of cultural conditions, such as pH, nitrogen source, substrate concentration, and temperature, were examined. Maximum production of C1 and CX cellulases occurred at 45 C in 2 to 4 days, in the presence of 1% Solka/Floc as substrate, when NaNO3 or urea used as sources of nitrogen. Under these conditions, cellulolytic activity of culture filtrates appeared to be similar to that reported for Trichoderma viride grown in a favorable environment. However, comparable yields of cellulase were produced by S. thermophile in less than one-quarter the time required by mesophilic fungi.     

Factors influencing the production of cellulases by Sporotrichum thermophile.

Cellulase production and growth of a strain of Sporotrichum thermophile were studied by using a mineral salts medium supplemented with yeast extract and insoluble cellulose. The effects of cultural conditions, such as pH, nitrogen source, substrate concentration, and temperature, were examined. Maximum production of C1 and CX cellulases occurred at 45 C in 2 to 4 days, in the presence of 1% Solka/Floc as substrate, when NaNO3 or urea used as sources of nitrogen. Under these conditions, cellulolytic activity of culture filtrates appeared to be similar to that reported for Trichoderma viride grown in a favorable environment. However, comparable yields of cellulase were produced by S. thermophile in less than one-quarter the time required by mesophilic fungi.     

养虾废水生物处理过程中营养负荷的动态变化

本文研究了几种方法对养虾废水的处理效果.在水体自净作用及投加翡翠贻贝、细基江蓠繁枝变种、活菌制剂和混合投加上述三种生物等5种处理条件下,测定了养殖废水中的氨、亚硝酸盐、硝酸盐、总氮、磷酸盐、总磷、溶解态COD、颗粒态COD、总COD、浮游植物总数、浮游动物总数和细菌总数等水质指标.结果表明:处理过程中各营养负荷变化趋势不同且相互之间有联系;各种处理对无机氮的减少有一定效果,并都使细菌总数明显下降;各处理不能降低COD含量,但可以改变COD形态;投加贻贝能明显抑制浮游植物的数量.     

Degradation of lignin in pulp mill wastewaters by white-rot fungi on biofilm

An investigation was conducted to explore the lignin-degrading capacity of attached-growth white-rot fungi. Five white-rot fungi, Phanerochaete chrysosporium, Pleurotus ostreatus, Lentinus edodes, Trametes versicolor and S22, grown on a porous plastic media, were individually used to treat black liquor from a pulp and paper mill. Over 71% of lignin and 48% of chemical oxygen demand (COD) were removed from the wastewater. Several factors, including pH, concentrations of carbon, nitrogen and trace elements in wastewater, all had significant effects on the degradation of lignin and the removal of COD. Three white-rot fungi, P. chrysosporium, P. ostreatus and S22, showed high capacity for lignin degradation at pH 9.0-11.0. The addition of 1 g l-1 glucose and 0.2 g l-1 ammonium tartrate was beneficial for the degradation of lignin by the white-rot fungi studied.     

Distillery and curd whey wastes as viable alternative sources for biosurfactant production

Biosurfactant production from synthetic medium and industrial waste, viz. distillery and whey wastes was investigated by using an oily sludge isolate Pseudomonas aeruginosa strain BS2. In synthetic medium separately supplemented with glucose and hexadecane as water-soluble and -insoluble carbon sources, respectively, strain BS2 reduced the surface tension of the fermentation broth from 57 to 27 mN/m. The culture produced biosurfactant during the stationary growth phase and its yield was 0.97 g/l. The culture utilized distillery and whey wastes for its growth, as maximum cell counts reached to 54 × 10⁸ and 64 × 10⁹ c.f.u./ml from an initial inoculum size of 1 × 0⁵ c.f.u./ml, respectively, within 48 h of incubation and in these wastes the yields of biosurfactant obtained were 0.91 and 0.92 g/l, respectively. In synthetic medium, distillery and whey wastes, strain BS2 produced a crystalline biosurfactant which belonged to the category of secondary metabolites and its maximum production occurred after the onset of nitrogen-limiting conditions. After recovering biosurfactant from the fermented waste, the chemical oxygen demand (COD) of distillery and whey wastes was significantly reduced by 81 and 87%, respectively. Total acids, nitrogen and phosphate levels in distillery waste were reduced by 90, 92 and 92%, respectively, while in case of whey waste the concentration of these nutrients was reduced by 88, 95 and 93%, respectively. The isolated biosurfactant possessed potent surface active properties, as it effectively reduced the surface tension of water from 72 to 27 mN/m and formed 100% stable emulsions of a variety of water-insoluble compounds such as hydrocarbons, viz. hexadecane, crude oil, kerosene and oily sludge and pesticides, viz. dichlorodiphenyltrichloroethane (DDT) and benzene hexachloride (BHC). The effectiveness of biosurfactant was also evident from its low critical micellar concentration (CMC) which was 0.028 mg/ml.     

Recognition and degradation of chitin by streptomycetes

Chitin is the second most abundant renewable polysaccharide, as it is a component of the exoskeleton of many organisms and of the cell walls of numerous fungi. Most streptomycetes secrete a number of chitinases, hydrolyzing chitin to oligomers, chitobiose or N-acetylglucosamine which can be utilized as carbon or nitrogen source. The chitinases of several streptomycetes have been shown to have a modular arrangement comprising catalytic, substrate binding as well as linker domains. Moreover, during growth in the presence of chitin-containing substrates, many Streptomycesstrains have been shown to secrete formerly unknown, small (about 200 aa) chitin binding proteins (CHBs) which lack enzymatic activity and specifically target and invade chitin. Several motifs, including the relative location and spacing of four tryptophan residues, are conserved in the investigated CHB types, CHB1 and CHB2. The affinity of CHB1 to crab shell chitin is two times higher than that of CHB2. Comparative studies of various generated mutant CHB1 proteins led to the conclusion that it is one of the exposed tryptophan residues that directly contributes to the interaction with chitin. On the basis of immunological, biochemical and physiological studies, it can be concluded that the CHBs act like a glue with which streptomycetes target chitin-containing samples or organisms. The ecological implications of these findings are discussed.     

有机碳源下废水厌氧氨氧化同步脱氮除碳

为明确有机碳源胁迫下,厌氧氨氧化反应器的同步脱氮除碳规律及功能微生物群落结构的动态变化,采用成功启动的厌氧氨氧化UASB反应器,通过逐步提升进水有机负荷,探究有机碳源下废水厌氧氨氧化同步脱氮除碳。研究表明,当进水化学需氧量(Chemical oxygen demand,COD)浓度从172 mg/L升至620 mg/L,反应器维持较高的脱氮效率,氨氮和总氮去除率均在85%以上,并对COD具有平均56.6%的去除率,高浓度COD未对Anammox菌活性构成显著抑制作用。聚合酶链式反应和变性梯度凝胶电泳(PCR-DGGE)图谱和割胶测序结果表明,变形菌门Proteobacteria、浮霉菌门Planctomycetes、绿曲挠菌门Chloroflexi以及绿菌门Chlorobi等微生物共存于同一反应体系中,推测反应器内存在复杂的脱氮除碳途径。而且,代表厌氧氨氧化的部分浮霉菌门微生物能耐受高浓度有机碳源,在高有机负荷下依旧发挥着高效的脱氮作用,为反应器高效脱氮提供了保障。     

Rapid assay for detection of microorganisms producing DNA-damaging metabolites.

Microorganisms producing DNA-damaging metabolites (i.e., fungi and streptomycetes) were detected by the Bacillus subtilis rec assay with agar plugs from plates on which the microorganisms had been grown. This assay allowed rapid identification of aflatoxinogenic fungi and streptomycetes producing strong DNA-damaging metabolites. For screening programs, several media have to be used to grow the microorganisms to be tested.     

Rapid assay for detection of microorganisms producing DNA-damaging metabolites

Microorganisms producing DNA-damaging metabolites (i.e., fungi and streptomycetes) were detected by the Bacillus subtilis rec assay with agar plugs from plates on which the microorganisms had been grown. This assay allowed rapid identification of aflatoxinogenic fungi and streptomycetes producing strong DNA-damaging metabolites. For screening programs, several media have to be used to grow the microorganisms to be tested.     

Reduction of pollutant parameters of leaf protein concentrate by-product by microbial fermentation

Deproteinised juice (DLJ) is left as a waste of a leaf protein concentrate (LPC) production plant. As it contains carbohydrates, nitrogenous compounds, lipids, minerals, and vitamins, its values of chemical oxygen demand (COD) and biological oxygen demand (BOD) are high enough to pose a serious disposal problem. But this waste, due to its nutrient content, is suitable for microbial growth. The growth of a number of filamentous fungi such as Rhizopus arrhizus, Aspergillus japonicus, and Trichoderma viride have been studied in different DLJ samples from different plants, such as cowpea, winged bean, radish, and turnip. The biomasses of fungi have been harvested and found to contain a large amount of protein. The fungi were found to reduce the high COD and BOD values of the DLJ samples, indicating the possibility of large scale use of this liquid waste as a fermentation medium.     

Using black soldier fly larvae for processing organic leachates

A large number of biodegradable byproducts including alcohols, soluble saccharides, volatile organic acids, and amines accumulate in the liquid fraction (leachate) produced as vegetal and food scrap waste decomposes. Untreated leachate, because it is rich in nutrients and organic byproducts, has a high chemical oxygen demand and is normally cleared of soluble organic byproducts by mineralization before its discharge into waterways. Mineralizing leachates using chemical and microbial biotechnologies is, however, a lengthy and costly process. We report here that the larvae of the black soldier fly Hermetia illucens (L.) (Diptera: Stratiomyidae), an insect rich in protein and lipids, and having significant commercial value, while feeding and growing off of compost leachate, lowers its chemical oxygen demand relative to that of leachate unexposed to larvae, neutralizes its acidity, and clears it of volatile organic acids, amines, and alcohols. These observations demonstrate that black soldier fly larvae could be used to help offset the cost and clean up of organic solutes in leachate waste streams while recycling carbon, nitrogen, and phosphate into usable and commercially valuable biomass.     

Utilization of Organic Nitrogen Sources by Two Phytoplankton Species and a Bacterial Isolate in Pure and Mixed Cultures

Algal production of dissolved organic carbon and the regeneration of nutrients from dissolved organic carbon by bacteria are important aspects of nutrient cycling in the sea, especially when inorganic nitrogen is limiting. Dissolved free amino acids are a major carbon source for bacteria and can be used by phytoplankton as a nitrogen source. We examined the interactions between the phytoplankton species Emiliania huxleyi and Thalassiosira pseudonana and a bacterial isolate from the North Sea. The organisms were cultured with eight different amino acids and a protein as the only nitrogen sources, in pure and mixed cultures. Of the two algae, only E. huxleyi was able to grow on amino acids. The bacterium MD1 used all substrates supplied, except serine. During growth of MD1 in pure culture, ammonium accumulated in the medium. Contrary to the expectation, the percentage of ammonium regenerated from the amino acids taken up showed no correlation with the substrate C/N ratio. In mixed culture, the algae grew well in those cultures in which the bacteria grew well. The bacterial yields (cell number) were also higher in mixed culture than in pure culture. In the cultures of MD1 and T. pseudonana, the increase in bacterial yield (number of cells) over that of the pure culture was comparable to the bacterial yield in mixed culture on a mineral medium. This result suggests that T. pseudonana excreted a more-or-less-constant amount of carbon. The bacterial yields in mixed cultures with E. huxleyi showed a smaller and less consistent difference than those of the pure cultures of MD1. It is possible that the ability of E. huxleyi to use amino acids influenced the bacterial yield. The results suggest that interactions between algae and bacteria influence the regeneration of nitrogen from organic carbon and that this influence differs from one species to another.     

Gelatinase Activity by Marine Fungi

Gelatinase activity was detectable in the culture filtrates of 13 of 14 species of marine fungi tested. The fungi were grown under conditions of submerged culture in a medium consisting of corn meal, soybean meal, and CaCO(3). The degree of enzymatic activity did not necessarily correlate with the amino nitrogen content of the filtrates. Eight of the filtrates contained measurable quantities of glutamic acid and alanine. Highest levels of gelatinase were obtained in the culture filtrates of Halosphaeria mediosetigera. Increasing the corn meal concentration of the medium resulted in even greater production of gelatinase by H. mediosetigera, whereas amino acid yields were enhanced by higher concentrations of soybean meal. Supplementation of the high corn meal medium with either a mineral mixture or distillers' solubles brought about a faster rate of production of gelatinase and, in the case of the distillers' solubles, a significantly higher level of enzyme, as well.