Investigation of keratinolytic and non-keratinolytic fungi grown above or below a 1-cm sewage sludge blanket

This study investigated the incidence of keratinolytic and non-keratinolytic fungi grown above or below a 1-cm sewage sludge blanket. The hair baiting method was used. Incubation was carried out at 23 and 37  °C. The number of keratinolytic fungi occurrences below a sludge blanket (anoxic conditions) was almost two times smaller than the number of fungal occurrences above this blanket (oxic conditions). The anoxic conditions did not significantly affect the number of non-keratinolytic fungi. Qualitative differences were also observed. Trichophyton ajelloi with its teleomorph Arthroderma uncinatum and some other fungi were found to prefer oxic conditions. In the case of non-keratinolytic fungi, the most evident differences were observed at 37  °C. Aspergillus fumigatus prevailed above a sludge blanket, while Pseudallescheria boydii was the predominating species below this blanket. The incidence of keratinolytic fungi was dependent on sludge physico-chemical characteristics: mainly on sludge organic matter stabilization (total nitrogen and ammonium nitrogen contents, proteolytic activity and C:N ratio) and hygienization (total and fecal coliforms) factors.     

Keratinolytic fungi in sewage sludge

Sewage sludge from the Upper Silesia Region of Poland were surveyed for keratinolytic fungi. Out of 100 Petri dishes examined, 89 were positive for these micro-organisms. Altogether, 185 fungal appearances belonging to 10 species were observed. Trichophyton terrestre with its teleomorph Arthroderma quadrifidum, T. ajelloi with A. uncinatum, Microsporum gypseum with Arthroderma sp., and Chrysosporium keratinophilum with Aphanoascus keratinophilus prevailed in the sludges. The sewage treatment technologies together with the sludge structure, humidity and pH were found to be critical factors determining the occurrence of keratinolytic fungi in the sludge environment. The qualitative and quantitative composition of keratinolytic fungi could be a useful tool in evaluation of sludge treatment processes.     

Removal of intl1 and associated antibiotics resistant genes in water,sewage sludge and livestock manure treatments

Antibiotics resistant genes (ARGs) are frequently detected in different media, such as wastewater, sewage sludge, and livestock manure. ARGs have been evidenced to have more and more threats to our environment because of their increase in species and total abundances causing more attention especially in horizontal gene transfer (HGT). The integron, an important form of mobile gene elements transfer ARGs through HGT, is demonstrated to have a high risk to human beings, and the class 1 integron (intl1), a predominant integron, is a marker of the process of horizontal gene transfer. Tetracycline, sulfonamides, macrolide, beta-lactam, trimethoprim, and quinolone plasmid-mediated resistance genes, which are frequently detected in various media and different treatment processes, have been reported to have significant correlations with intl1. Considering the risk of ARGs, especially those associated with intl1, the removal of intl1 and associated ARGs from water, sludge and livestock manure has attracted much more attention recently. Therefore, the mechanism and removal of intl1 and associated ARGs in water depth treatment, sludge and livestock manure digestion and composting process are reviewed in this paper. Besides, the limitation of the current study in this field is discussed, and the key points in the future investigation are provided.     

Degradation of keratin substrates by fungi isolated from sewage sludge

Four fungal species including two dermatophytes and two saprophytes were isolated from sewage sludge samples at Basrah (Iraq) they were tested for their degradative ability towards three types of keratin substrates (human hair, chicken feathers and wool). The rate of keratin degradation was expressed as weight loss over three weeks of incubation using a liquid culture medium. Human hair had the highest degradation rate by colonization of Chrysosporium pannicola and Microsporum gypseum at a rate of 62% and 4% respectively. Chicken feathers were highly degraded by Aspergillus flavus (32%) while wool degradation was highest by C. pannicola (45.5%) and Trichophyton mentagrophytes var. erinacei (38%). There was a significant difference (p < 0.00l) in keratin substrate degradation rates by the examined fungi. Keratinase activity was highest for C. pannicola and M. gypseum in the culture medium baited with human hair. Aspergillus flavus revealed the highest activity of this enzyme in cultures amended with chicken feathers while T. mentagrophytes var. erinacei showed highest keratinase activity in cultures with wool substrate. The amount of protein released into the culture medium varied among the tested fungi. The medium's alkalinity increased over incubation time from 6.5 to 7.8. Microscopic examination showed maceration of the keratin substrates by the fungi. This revised version was published online in June 2006 with corrections to the Cover Date.     

Influence of bulking agent on sewage sludge composting process

Four types of compost, consisting of mixtures of Acacia dealbata (A) with sewage sludge (SS) were studied in a laboratory reactor. Composting time was 80 days and parameters monitored over this period included temperature, organic matter, pH, CO₂, O₂, C/N ratio, Kjeldahl-N, as well as maturity indexes. All the studied parameters were influenced by the bulking amount used. The highest profile temperature measured was for the A/SS 1/2 (w/w) mixture that reached a maxima temperature of 67 °C and lower maximum temperatures of 52, 48 and 46 °C were observed for A/SS 1/3, 1/1 and 1/0 composts, respectively. The kinetic model used showed that a descent of sewage sludge in the composting mixtures favored the enzyme–substrate affinity. However, an increase in depending on the parameters of the process factors was observed when the sewage sludge ratio was increased in mixtures. The optimal amounts of sewage sludge for co-composting with Acacia indicate that moderate amounts of sludge (1/1) would be the best compromise.     

Plasma ammonia and urea levels in Atlantic salmon farmed in sea water

Plasma ammonia and urea levels were measured in feeding Atlantic salmon Salmo salar L. reared in two different commercial Norwegian sea farms. A very large individual variation of the plasma ammonia and urea levels in addition to the large influence of several factors suggest that it may be difficult to use these parameters in diagnostic work on fish.     

Human enteropathogen load in activated sewage sludge and corresponding sewage sludge end products

This study demonstrated a significant reduction in the concentrations of Cryptosporidium parvum and Cryptosporidium hominis oocysts, Giardia lamblia cysts, and spores of human-virulent microsporidia in dewatered and biologically stabilized sewage sludge cake end products compared to those of the respective pathogens in the corresponding samples collected during the sludge activation process.     

The effect of sewage sludge composting on the quantitative state of some groups of bacteria and fungi

Analysis of the quantitative state of disease causing bacteria and of other microbic groups were done on the sewage sludge from a sewage treatment plant. The results of the analysis include the ammonifying bacteria, nitrifying and denitrifying bacteria. The general quantity of bacteria and fungi in a secondary dehydrated sludge, fermented secondary dehydrated sludge, and in composted secondary dehydrated sludge was deterinated. Composts were prepared from dehydrated secondary sludge with the addition of sawdust. Microbiological analysis of sewage sludge showed, that the quantities of the fecal coli bacteria were 6500; 220 and 150 cells per cm3 of the secondary dehydrated sludge, fermented secondary dehydrated sludge and composted dehydrated secondary sludge, respectively. The numbers of Salmonella were respectively 67.80; 6.48 and 6.60 cells per cm3. The general numbers of bacteria were 2.98 x 10(7); 2.79 x 10(7); 2.15 x 10(7) cells per cm3 of sludge. The cell numbers of fungi were: 6.20x 10(2); 19.60 x 10(2); 7.80 x 10(2) per cm3 of sludge. In the three types of sludge, the results show great numbers of the ammonifying, nitrifying and denitrifying bacteria. Of the analysed groups of bacteria, the highest numbers of cells were found for general bacteria; ammonifying and nitrifying bacteria were next in abundance; still fewer were the denitrifying bacteria. Fungi and pathogenic bacteria were the least numerous.     

Molecular evolution of urea amidolyase and urea carboxylase in fungi

Background  

Urea amidolyase breaks down urea into ammonia and carbon dioxide in a two-step process, while another enzyme, urease, does this in a one step-process. Urea amidolyase has been found only in some fungal species among eukaryotes. It contains two major domains: the amidase and urea carboxylase domains. A shorter form of urea amidolyase is known as urea carboxylase and has no amidase domain. Eukaryotic urea carboxylase has been found only in several fungal species and green algae. In order to elucidate the evolutionary origin of urea amidolyase and urea carboxylase, we studied the distribution of urea amidolyase, urea carboxylase, as well as other proteins including urease, across kingdoms.     

Influence of humic substances on bioavailability of Cu and Zn during sewage sludge composting

Influence of humic substances (HS) on bioavailability of Cu and Zn was characterized during 120 days co-composting of sewage sludge and maize straw. At the initial stage of composting, Cu and Zn in sewage sludge were released as organic matter was degraded, and water soluble Cu and Zn increased markedly. Water soluble Cu and FA content decreased after 21 days whereas water soluble Zn increased during the whole process. Both HA-Cu and HA-Zn were significantly and positively correlated with HA and H/F, respectively. At the end of composting, the distribution coefficients of HA-Cu and HA-Zn reached 27.50% and 3.33% respectively with HA-Cu/HA-Zn ratio increased from 1.29 to 2.73. The results suggest that Cu combined with HA more strongly than Zn, and composting treatment could decrease bioavailability of Cu markedly.     

The time of urea treatment and its effects on the succession of ammonia fungi in two warm temperate forests in Japan

We studied the effects of the timing of urea treatment on the succession of ammonia fungi. In two evergreen Castanopsis cuspidata forests and in one deciduous Quercus serrata forest, we applied 343g urea to 25 and 15 plots of 0.5m², respectively, at three different times of the year. Ten of the early-phase (EP) species, considered to be saprotrophic, and 6 of the late-phase (LP) ones, considered ectomycorrhizal, fruited. In both phases, the commencement, peak, and cessation of fruiting took place simultaneously among all the plots treated at the same time. The fruiting occurred in summer and autumn. Quantity and size of the fruit bodies was larger in the LP than in the EP species. Fruiting of EP species was affected by the treatment time and that of LP species by interaction of the treatment time and vegetation type. EP was short and occurred as one period, whereas LP was long and occurred as two or more fruiting seasons. We found that species composition, dominant species, and degree of its dominance in fruiting of the ammonia fungi are predictable for different treatment times of the year and different vegetation types.     

Efficient,high-speed methane fermentation for sewage sludge using subcritical water hydrolysis as pretreatment

A novel biomass-energy process for the production of methane from sewage sludge using a subcritical water (sub-CW) hydrolysis reaction as pretreatment is proposed. The main substances of sewage sludge hydrolyzed by sub-CW at 513 K for 10 min were acetic acid, formic acid, pyroglutamic acid, alanine, and glycine. Fermentation experiments were conducted in an anaerobic-sludge reactor for two different samples: real sewage sludge and a model solution containing components typically produced by the sub-CW pretreatment of sewage sludge. In the experiment for the sub-CW pretreatment of sewage sludge, methane generation was twice that for non-pretreatment after 3 days of incubation. In the model experiment, the methane conversion was about 40% with the application of mixture of organic acids and amino acids after 5 days of incubation. Furthermore, the methane conversion was about 60% for 2 days when only organic acids, such as acetic acid and formic acid, were applied. Because acetic acid is the key intermediate and main precursor of the methanogenesis step, fermentation experiments were conducted in an anaerobic-sludge reactor with high concentrations of acetic acid (0.01–0.1 M). Nearly 100% of acetic acid was converted to methane and carbon dioxide in 1–3 days.     

土壤盐渍化对尿素与磷酸脲氨挥发的影响

氨挥发是肥料氮素损失的重要途径之一,肥料类型、土壤类型、肥料用量以及土壤全盐量均影响氨挥发损失率及挥发特征。本文采用通气法测定了磷酸脲和尿素两种肥料六个施肥量处理分别施入六个不同盐渍化程度（1.7、9.9、16.4、23.2、29.1、37.9 g/kg）的土壤后氨挥发累积状况和动力学特性,以及土壤氨挥发累积量与土壤电导值之间的相关性。结果表明：（1）在土壤总盐介于1.66 -37.9 g/kg的范围内,随着土壤含盐量增加,尿素与磷酸脲处理的氨挥发累积量显著增加;土壤含盐量对氨挥发速率有显著的促进作用。（2）各处理二次线性函数拟合的二项式系数a均为负值,表明：在不同盐渍化条件下肥料的挥发速率是随着时间增长而降低的;一次线性函数和Elovich 方程的斜率a随土壤含盐量增加而增大,表明：土壤盐渍化将加剧土壤的氨挥发速率。（3）土壤氨挥发累积量与电导值拟合结果符合logistic方程（︱R︱分别为0.9732,0.9815,0.965,0.9182,0.9817,0.9971︱R︱>r0.01=0.9172, n=6）,氨挥发累积量随土壤电导值呈“S”型增长。     

Characterization of anaerobic non-spore-forming bacteria in municipal sewage sludge and integrated paper-mill waste water

Summary Forty-three isolates of Gram-negative, mesophilic, non-spore-forming anaerobic cellulolytic bacteria were obtained from (i) an anaerobic reactor treating waste water from an integrated paper mill and (ii) an anaerobic sewage-sludge digestor. These isolates were studied for carbohydrate fermentation and fermentation products. By numerical techniques, 22 isolates could be placed in two groups: group A (10 isolates) and group B (12 isolates). The isolates belonging to group A showed degradation of filter paper in 2–7 days. They were slightly-curved long rods and similar toBacteroides cellulosolvens andAcetivibrio cellulolyticus. Acetic acid was produced as major product. The bacteria also produced ethanol, isobutanol, pyruvic and lactic acids. Group B strains degraded filter paper in 4–5 weeks. They were short rods and produced propionic, lactic, succinic and acetic acids as fermentation products. The remaining 21 isolates could disintegrate filter paper in 2–5 weeks. They showed variable fermentation patterns, both as to fermentable carbohydrates and end products. Except for one isolate, which showed obvious similarity toButyrivibrio fibrisolvens, the isolate differed distinctly from reference strains of ruminai origin.

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Caractérisation des bactéries anaérobies non-sporulantes dans la boue résiduaire de station d'épuration des eaux domestiques (boues d'égout) et dans l'eau résiduaire d'une usine intégrale de pâte à papier

Résumé 43 souches de bactéries cellulolytiques Gram-négatives, mésophiles, non-sporulantes, et anaérobies ont été isolées à partir (A) d'un réacteur anaérobie traitant l'eau résiduaire d'une usine intégrale de pâte à papier et (B) de digesteurs anaérobies de boues d'égout. Ces souches ont été étudiées quant à leurs propriétés de fermenter les hydrates de carbone et quant à leurs produits de fermentation. Des techniques numériques ont permis de classer 22 de ces souches dans deux groupes: le groupe A (10 souches) et le groupe B (12 souches). Les souches appartenant au groupe A dégradaient le papier filtre en 2 à 7 jours. Elles se présentaient sous la forme de longs bâtonnets curvilignes et ressemblaient àBacteroides cellulosolvens et àAcetivibrio cellulolyticus. Le metabolite majeur était l'acide acétique. Elles produisaient aussi de l'éthanol, de l'iso-butanol, et des acides pyruvique et lactique. Les souches appartenant au groupe B dégradaient le papier filtre en 4 à 5 semaines. Elles se présentaient sous la forme de bâtonnets courts et produisaient, comme métabolites, les acides propionique, lactique, succinique et acétique. Les 21 souches restantes dégradaient le papier filtre en 2 à 5 semaines. Elles présentaient des profils de fermentation variables tant en ce qui concernait les hydrates de carbone fermentés que les produits finaux. A part une souche qui présentait une similitude évidente avecButyrivibrio fibrisolvens, ces souches différaient de façon marquée d'avec les souches de référence du rumen.

     

Inhibition of methanogenesis in pure cultures by ammonia, fatty acids, and heavy metals, and protection against heavy metal toxicity by sewage sludge

The effect of ammonium chloride, sodium butyrate, sodium propionate, and the heavy metals nickel, zinc, and copper on methanogenesis by pure cultures of Methanospirillum hungatei, Methanosarcina barkeri, Methanobacterium thermoautotrophicum, and Methanobacterium formicicum at pH 6.5 was studied. The latter three strains were resistant to greater than 60 g/L of the volatile fatty acids and to greater than 10 g/L of NH3 N. Methanospirillum hungatei was somewhat more sensitive with 50% inhibition of methanogenesis occurring at 4.2 g/L NH3 N, 27 g/L butyrate, and 41 g/L propionate. All strains were very sensitive to both copper (1-5 mg/L) and zinc (1-10 mg/L), but much more resistant to nickel. Zinc and copper concentrations 30 to 270 times higher were required to cause inhibition of Msp. hungatei incubated in sewage sludge compared with buffer, indicating a strong protective environment was afforded the methanogens against heavy metal toxicity in the sludge.     

Occurrence, growth, and suppression of salmonellae in composted sewage sludge

Composted sewage sludge may be used to improve soil quality, but there remains some doubt concerning the microbiological safety of the product. Sewage sludge composts from 30 municipalities were sampled, and four samples (12%) contained salmonellae (two contained fewer than 0.3/g, and the other two had 21/g and 1.7 X 10(4)/g). All 30 composts were inoculated with salmonellae; the populations decreased at a specific death rate of about 0.15 h-1 over 24 h at 36 degrees C. In irradiation-sterilized composts inoculated with salmonellae, the salmonellae grew at a rate of 0.65 doublings per h for over 24 h. Growth and death rates were found to be moisture and flora associated. The growth or death rates for antibiotic-resistant salmonellae were not different from those of nonresistant strains. It was concluded that the active indigenous flora of compost establishes a homeostatic barrier to colonization by salmonellae, and in the absence of competing flora, reinoculated salmonellae may grow to potentially hazardous densities. The active microflora of moist composts eliminated contaminating salmonellae (10(5)/g) after 6 weeks.     

Long-term effects of metals in sewage sludge on soils,microorganisms and plants

Summary This paper reviews the evidence for impacts of metals on the growth of selected plants and on the effects of metals on soil microbial activity and soil fertility in the long-term. Less is known about adverse long-term effects of metals on soil microorganisms than on crop yields and metal uptake. This is not surprising, since the effects of metals added to soils in sewage sludge are difficult to assess, and few long-term experiments exist. Controlled field experiments with sewage sludges exist in the UK, Sweden, Germany and the USA and the data presented here are from these long-term field experiments only. Microbial activity and populations of cyanobacteria,Rhizobium leguminosarum bv.trifolii, mycorrhizae and the total microbial biomass have been adversely affected by metal concentrations which, in some cases, are below the European Community's maximum allowable concentration limits for metals in sludge-treated soils. For example, N₂-fixation by free living heterotrophic bacteria was found to be inhibited at soil metal concentrations of (mg kg^–1): 127 Zn, 37 Cu, 21 Ni, 3.4 Cd, 52 Cr and 71 Pb. N₂-fixation by free-living cyanobacteria was reduced by 50% at metal concentrations of (mg kg^–1): 114 Zn, 33 Cu, 17 Ni, 2.9 Cd, 80 Cr and 40 Pb.Rhizobium leguminosarum bv.trifolii numbers decreased by several orders of magnitude at soil metal concentrations of (mg kg^–1): 130–200 Zn, 27–48 Cu, 11–15 Ni, and 0.8–1.0 Cd. Soil texture and pH were found to influence the concentrations at which toxicity occurred to both microorganisms and plants. Higher pH, and increased contents of clay and organic carbon reduced metal toxicity considerably. The evidence suggests that adverse effects on soil microbial parameters were generally found at surpringly modest concentrations of metals in soils. It is concluded that prevention of adverse effects on soil microbial processes and ultimately soil fertility, should be a factor which influences soil protection legislation.