Anaerobic microflora of everglades sediments: effects of nutrients on population profiles and activities.

Everglades sediments (wetland soils) near sources of agricultural runoff had low redox potentials, were blackened with sulfide, and displayed high porewater phosphorus (total) concentrations and high water column conductivities. These sediments yielded 10(sup3)- to 10(sup4)-fold-higher numbers of culturable anaerobes, including methanogens, sulfate reducers, and acetate producers, than did sediments from Everglades and Lake Okeechobee comparative control sites not as directly associated with agricultural runoff. These observations demonstrated that there was a general, rather than specific, enhancement of the anaerobic microflora in the sediments most likely influenced by agricultural runoff. Despite these differences in microfloral patterns, methylmercury and total mercury levels were similar among these contrasting sediments. Although available sulfate and phosphorus appeared to stimulate the productivity of sulfate reducers in Everglades sediments, the number of culturable sulfate reducers did not directly correspond to the concentration of sulfate and phosphorus in porewaters. Microcosms supplemented with sulfate, nitrate, and phosphate altered the initial capacities of the sediment microflora to produce acetate and methane from endogenous matter. For sediments nearest sources of agricultural runoff, phosphorus temporarily enhanced acetate formation and initially suppressed methane production, sulfate enhanced acetate formation but did not significantly alter the production of methane, and nitrate totally suppressed the initial production of both methane and acetate. In regards to the latter, microbes capable of dissimilating nitrate to ammonium were present in greater culturable numbers than denitrifiers. In microcosms, acetate was a major source of methane, and supplemental hydrogen was directed towards the synthesis of acetate via CO(inf2)-dependent acetogenesis. These findings demonstrate that Everglades sediments nearest agricultural runoff have enhanced anaerobic microbial profiles and that the anaerobic microflora are poised to respond rapidly to phosphate, sulfate, and nitrate input.

Ammonia-induced cell envelope injury in Escherichia coli and Enterobacter aerogenes.

Ammonia-induced cell envelope injury was examined in pure cultures of Escherichia coli and Enterobacter aerogenes. Cell injury, as determined by the ratio of colony-forming units on m-T7 agar to colony-forming units on m-Endo agar, increased with exposure to increasing concentrations of ammonia. Cell envelopes appeared to be the site of injury as indicated by increasing susceptibility to lysozyme with increasing ammonia concentration. Cells exposed to ammonia also exhibited more cellular leakage than control cells. Leakage from cells exposed to ammonia included proteins, and all leaked substances increased in concentration as ammonia concentrations increased. The concentration of 2-keto-3-deoxyoctonate (KDO) in the outer membrane of E. coli increased with ammonia exposure, while KDO concentration in the outer membrane of E. aerogenes decreased. The results suggest that exposure of E. coli cells to high concentrations of ammonia disrupts the outer membrane and lipopolysaccharide-associated proteins, while E. aerogenes cells are affected through the disruption of bonds between KDO and the outer membrane.

Ammonia-induced injury in pure cultures and natural populations of coliform bacteria.

Ammonia-induced injury was investigated in pure cultures of Escherichia coli and Enterobacter aerogenes, and in natural coliform populations obtained from the oligotrophic Luxapallila and the eutrophic Sunflower Rivers in northern Mississippi. Pure cultures were affected by ammonia exposure as indicated by changes in the injury ratio (IR) of CFU on m-T7 agar/CFU on m-Endo agar. Ammonia concentrations between 0 and 20 (mg NH3-N/1) had little or no effect and concentrations between 40 and 80 caused the greatest injury. Natural coliform populations from the oligotrophic river were more prone to ammonia-induced injury than those from the eutrophic river. The results stress the need for the routine use of m-T7 media and the enumeration of injured cells when using the membrane filter procedure to ascertain domestic water quality.

Impact of nitrogen and phosphorus on [C]lignocellulose decomposition by stream wood microflora.

Nutritional and physical factors affecting the decomposition of [C]lignocellulose prepared from Douglas fir (Pseudotsuga menziesii) were examined by incubating the labeled substrate with homogenized surface wood scrapings obtained from a Douglas fir log in a Pacific Northwest stream. Incubations were conducted in distilled water, in stream water collected from four different sources, or in a defined mineral salts solution with or without supplemental N (KNO(3)). Decomposition rates of [C]lignocellulose, as measured by CO(2) evolution, were greater in each of the four filter-sterilized sources of stream water than in distilled water alone. Decomposition experiments conducted in stream water media with the addition of defined mineral salts demonstrated that [C]cellulose decomposition was stimulated 50% by the addition of either KNO(3) or KH(2)PO(4)/K(2)HPO(4) and further enhanced (167%) by a combination of both. In contrast, [C]lignin decomposition was stimulated (65%) only by the addition of both N and P. Decomposition of [C]lignocellulose was greatest when supplemental KNO(3) was supplied in concentrations of at least 10.0 mg of N liter but not increased further by higher concentrations. The decomposition of [C]lignocellulose increased as the incubation temperature was raised and NO(3)-N supplementation further increased these rates between three-and sevenfold over the range of temperatures examined (5 to 22 degrees C). Accumulation of NH(4) (2 to 4 mg of N liter) was always observed in culture filtrates of incubations which had been supplemented with KNO(3), the quantity being independent of NO(3) concentrations >/= 10 mg of N liter. The role of supplemental NO(3) in the decomposition of [C]lignocellulose is discussed in relation to wood decomposition and the low concentrations of N found in stream ecosystems of the Pacific Northwest.

Nitrogen dynamics in stream wood samples incubated with [C]lignocellulose and potassium [N]nitrate.

Surface wood samples obtained from a Douglas fir log (Pseudotsuga menziesii) in a Pacific Northwest stream were incubated in vitro with [C]lignocellulose in a defined mineral salts medium supplemented with 10 mg of N liter of N-labeled NO(3) (50 atom% N). Evolution of CO(2), distribution and isotopic dilution of N, filtrate N concentrations, and the rates of denitrification, N(2) fixation, and respiration were measured at 6, 12, and 18 days of incubation. The organic N content of the lignocellulose-wood sample mixture had increased from 132 mug of N to a maximum of 231 mug of N per treatment after 6 days of incubation. Rates of [C]lignocellulose decomposition were greatest during the first 6 days and then began to decline over the remaining 12 days. Total CO(2) evolution was also highest at day 6 and declined steadily over the remaining duration of the incubation. Filtrate NH(4)-N increased from background levels to a final value of 57 mug of N per treatment. Filtrate NO(3) N completely disappeared by day 6, and organic N showed a slight decline between days 12 and 18. The majority of the N that could be recovered appeared in the particulate organic fraction by day 6 (41 mug of N), and the filtrate NH(4) N fraction contained 11 mug of N by day 18. The N enrichment values of the filtrate NH(4) and the inorganic N associated with the particulate fraction had increased to approximately 20 atom% N by 18 days of incubation, whereas the particulate organic fraction reached its highest enrichment by day 6. Measurements of N(2) fixation and denitrification indicated an insignificant gain or loss of N from the experimental system by these processes. The data show that woody debris in stream ecosystems might function as a rapid and efficient sink for exogenous N, resulting in stimulation of wood decomposition and subsequent activation of other N cycling processes.

Microbial decomposition of wood in streams: distribution of microflora and factors affecting [C]lignocellulose mineralization.

The distribution and lignocellulolytic activity of the microbial community was determined on a large log of Douglas fir (Pseudotsuga menziesii) in a Pacific Northwest stream. Scanning electron microscopy, plate counts, and degradation of [C]lignocelluloses prepared from Douglas fir and incubated with samples of wood taken from the surface and within the log revealed that most of the microbial colonization and lignocellulose-degrading activity occurred on the surface. Labeled lignocellulose and surface wood samples were incubated in vitro with nutrient supplements to determine potential limiting factors of [C]lignocellulose degradation. Incubations carried out in a nitrogenless mineral salts and trace elements solution were no more favorable to degradation than those carried out in distilled water alone. Incubations supplemented with either (NH(4))(2)SO(4) or organic nitrogen sources showed large increases in the rates of mineralization over incubations with mineral salts and trace elements alone, with the greatest effect being observed from an addition of (NH(4))(2)SO(4). Subsequent incubations with (NH(4))(2)SO(4), KNO(3), and NH(4)NO(3) revealed that KNO(3) was the most favorable for lignin degradation, whereas all three supplements were equally favorable for cellulose degradation. Supplementation with glucose repressed both lignin and cellulose mineralization. The results reported in this study indicate that nitrogen limitation of wood decomposition may exist in streams of the Pacific Northwest. The radiotracer technique was shown to be a sensitive and useful tool for assessing relative patterns of lignocellulose decay and microbial activity in wood, along with the importance of thoroughly characterizing the experimental system before its general acceptance.

Microbial succession on a chitinous substrate in a woodland stream.

Plate counts, scanning electron microscopy, and direct observations were utilized to determine successional trends of different groups of microorganisms onProcambarus versutus (Hagen) exoskeletons incubatedin situ in a second-order, acidic woodland stream in Northwest Florida. Plate counts and SEM observations indicated a definite successional pattern dominated in numbers initially by the nonbranching bacteria, followed by the actinomycetes. The greatest number of fungal propagules coincided with the least number of bacterial colony-forming units. Chitinoclastic bacterial colonizers increased on the substrate throughout the study, comprising as much as 88% of the total bacterial community during the final stages of chitin decomposition. Scanning electron micrographs taken over the duration of the study revealed the abundance of an actinomycete, identified as a species ofStreptomyces, on the exoskeleton. Enumeration of microorganisms in the stream sediment was also performed in conjunction with the exoskeleton analysis. Relatively constant numbers of bacteria and fungi were observed, with chitinoclastic bacteria comprising between 3 and 11% of the total sediment bacterial community.