Nitrogen-Fixing and Uricolytic Bacteria Associated with the Gut of Dendroctonus rhizophagus and Dendroctonus valens (Curculionidae: Scolytinae)

The bark beetles of the genus Dendroctonus feed on phloem that is a nitrogen-limited source. Nitrogen fixation and nitrogen recycling may compensate or alleviate such a limitation, and beetle-associated bacteria capable of such processes were identified. Raoultella terrigena, a diazotrophic bacteria present in the gut of Dendroctonus rhizophagus and D. valens, exhibited high acetylene reduction activity in vitro with different carbon sources, and its nifH and nifD genes were sequenced. Bacteria able to recycle uric acid were Pseudomonas fluorescens DVL3A that used it as carbon and nitrogen source, Serratia proteomaculans 2A CDF and Rahnella aquatilis 6-DR that used uric acid as sole nitrogen source. Also, this is the first report about the uric acid content in whole eggs, larvae, and adults (male and female) samples of the red turpentine beetle (Dendroctonus valens). Our results suggest that the gut bacteria of these bark beetles could contribute to insect N balance.     

Nitrogen Requirements of Microtus rossiaemeridionalis

Total nitrogen requirements in the small herbivorous rodent Microtus rossiaemeridionaliswere studied. When provided with two kinds of standard laboratory food (about 20% and 0% digestible protein, respectively) ad libitum(cafeteria diet), the voles chose a diet of 14% protein. The total daily (with urine and faeces) nitrogen losses were, in this case, determined as 3.65 ± 0. 60 mg N/ g of body mass. When provided with a protein-free diet alone, the animals lost 0.90 ± 0,05 mg N/ g body mass per day, comprising about 3% of their body mass daily. It was proposed that the consumption of a low-fiber diet (about 4% in both kinds of food) could affect the gastrointestinal bacterial flora and, thus, lead to changes in the primarily herbivorous nitrogen metabolism.     

Impacts of Organic and Conventional Crop Management on Diversity and Activity of Free-Living Nitrogen Fixing Bacteria and Total Bacteria Are Subsidiary to Temporal Effects

A three year field study (2007–2009) of the diversity and numbers of the total and metabolically active free-living diazotophic bacteria and total bacterial communities in organic and conventionally managed agricultural soil was conducted using the Nafferton Factorial Systems Comparison (NFSC) study, in northeast England. Fertility management appeared to have little impact on both diazotrophic and total bacterial communities. However, copy numbers of the nifH gene did appear to be negatively impacted by conventional crop protection measures across all years suggesting diazotrophs may be particularly sensitive to pesticides. Impacts of crop management were greatly overshadowed by the influence of temporal effects with diazotrophic communities changing on a year by year basis and from season to season. Quantitative analyses using qPCR of each community indicated that metabolically active diazotrophs were highest in year 1 but the population significantly declined in year 2 before recovering somewhat in the final year. The total bacterial population in contrast increased significantly each year. It appeared that the dominant drivers of qualitative and quantitative changes in both communities were annual and seasonal effects. Moreover, regression analyses showed activity of both communities was significantly affected by soil temperature and climatic conditions.     

Volatile Fatty Acid Requirements of Cellulolytic Rumen Bacteria

A gas chromatographic method was developed which could separate the isomers isovaleric and 2-methylbutyric acid. Subsequent analyses revealed that most commercially available samples of these acids were cross-contaminated; however, one sample of each acid was found to be pure by this criterion. The growth response of seven strains of cellulolytic rumen bacteria (three strains of Bacteroides succinogenes, three strains of Ruminococcus flavefaciens, and one strain of R. albus) to additions of isobutyric, isovaleric, 2-methylbutyric, valeric, and combinations of valeric and a branched-chain acid was determined. Strains of B. succinogenes required a combination of valeric plus either isobutyric or 2-methylbutyric acid. Isovaleric acid was completely inactive. Either isobutyric or 2-methylbutyric acid was required for the growth of R. albus 7. Strain C-94 of R. flavefaciens grew slowly in the presence of any one of the three branched-chain acids, but a combination of isobutyric and 2-methylbutyric acids appeared to satisfy this organism's growth requirements. None of the individual acids or mixtures of straight- and branched-chain acids allowed growth of R. flavefaciens strain C1a which would approach the response obtained from the total mixture of acids. Further work indicated that all three branched-chain acids were required for optimal growth by this strain, although isovaleric acid only influenced the rate of maximal growth. Either 2-methylbutyric or isovaleric acid allowed growth of nearly the same magnitude as that of the positive control for R. flavefaciens B34b. The presence of acetic acid had little influence on the rate or extent of growth of any of the strains except R. albus 7, for which the extent of growth was markedly increased. Determination of the quantitative fatty acid requirements for the three B. succinogenes strains indicated that 0.1 μmole of valeric per ml and 0.05 μmole of 2-methylbutyric per ml permitted maximal growth. However, with isobutyric acid as the branched-chain component, strains A3c and B21a required 0.1 μmole/ml in contrast to S-85 which exhibited optimal growth at the 0.05 μmole/ml level. By use of mixtures of isobutyric and 2-methylbutyric acids, good growth of C-94 was obtained at concentrations of 0.1 and 0.01 μmole/ml, respectively. About 0.3 μmole/ml of each acid was required for satisfactory growth of C1a.     

Carbon and Nitrogen Content of Natural Planktonic Bacteria

A method of estimating carbon and nitrogen content per unit of natural bacterial cell volume was developed. This method is based on the difference in the retentiveness of bacteria between two kinds of glass fiber filter, GF/C and GF/F (Whatman, Inc., Clifton, N.J.). Biovolume and biomass (carbon and nitrogen content) of bacteria which passed through the GF/C but not the GF/F filter were estimated with an epifluorescence microscopy and a CHN analyzer, respectively. From seasonal determinations of natural planktonic bacteria in epilimnetic waters of a mesotrophic lake, the conversion factors of 106 fg of C/μm³ and 25 fg of N/μm³ were derived as average values. By using these values, the contribution of bacteria to the biomass of lake plankton is discussed.     

Nitrogen Fixation by Marine Photosynthetic Bacteria

S ummary . Potential nitrogen-fixing marine photosynthetic bacteria isolated from the coastal waters and sea-bed sediments of Cardigan Bay and adjoining mud flats, were investigated, using the acetylene reduction technique, to ascertain the magnitude of their contribution of fixed nitrogen to the coastal ecosystem. Aberystwyth harbour and Iceland fjord mud readily yielded Athiorhodaceae, the majority of which consistently reduced acetylene when grown as pure cultures on marine media. Only one such strain was isolated from local seawater, but they were more common in Iceland fjord water. No marine Thiorhodaceae were isolated. It was essential to monitor crude and pure culture systems for ethylene production in the absence of acetylene.     

甘蔗器官固氮酶活性及其对接种固氮菌的响应

以巴西固氮甘蔗品种B1、B8和广西主栽甘蔗品种ROC22和ROC16为材料,通过桶栽砂培方法,研究不同品种甘蔗及接种固氮菌R16SH后不同生育期和不同器官(根、茎、叶片、叶鞘)的固氮酶活性.结果显示,4个品种甘蔗及接菌后各器官的固氮酶活性均在伸长初期或伸长盛期达到高峰,且以根和茎较高,叶、叶鞘次之;B1、B8根的接菌效果最佳,其固氮酶活性分别比对照增加13.22%和12.42%,而B1和ROC16茎的固氮酶活性分别比对照提高12.19%和12.02%,叶片提高了8%~9%,叶鞘的固氮酶活性变化最小;4个品种甘蔗固氮酶活性表现为B8>B1>ROC22>ROC16,且品种B8与ROC22和ROC16存在显著或极显著差异;相对于品种ROC22和ROC16,固氮菌R16SH对B1与B8的株高以及B1的茎径有更好的生长诱导效应,且在伸长期表现最明显.研究表明,甘蔗的固氮酶活性在品种、器官及生育期间均存在明显差异,接种固氮菌能不同程度提高各品种器官的固氮酶活性,且各品种根、茎以及B1和B8的叶片固氮酶活性显著或极显著差异增加;巴西固氮甘蔗品种表现出比广西主栽甘蔗品种更强的固氮酶活性,且对接种固氮菌反应更敏感.     

Collagenolytic Activity of Some Marine Bacteria

Reconstituted, acid-extracted collagen was used to prepare a medium to screen proteolytic marine bacteria for their ability to elaborate collagenolytic enzymes. The medium was resistant to solubilization by trypsin, hyaluronidase, chondroitinase ABC, and various marine proteinases, but was readily hydrolyzed by commercial Clostridium collagenases. Eighty-seven marine isolates collected in the vicinity of Bermuda, Oahu (Hawaii), and Stone Harbor and Cape May, N. J., were screened. Approximately 44 per cent of the isolates were capable of elaborating enzymes that hydrolyzed reconstituted collagen gels. Several cultures produced collagenolytic enzymes only when grown in the presence of collagen or degradation products of collagen, and with very few exceptions the presence of collagen in the medium greatly enhanced collagenolytic enzyme production. The enzymes from a collagenolytic Bermuda marine isolate were studied in more detail to illustrate that the enzymes capable of hydrolyzing reconstituted collagen were separable from nonspecific proteinases by zone electrophoresis and that these enzymes were true collagenases by virtue of their ability to hydrolyze native bovine Achilles'tendon obtained from three different sources.     

Heterotrophic Activity of Deep-Sea Sediment Bacteria

Sediment samples, containing mixed microbial populations that were decompressed during retrieval from 7,750 and 8,130 m in the Puerto Rican Trench, were recompressed and incubated at the approximate in situ temperature (3 C) and pressure (775 or 815 atm) in the presence of 14C-labeled amino acids. Heterotrophic activity (total uptake, CO2 respiration, and cellular assimilation) and cellular-associated "pool" concentrations were measured. Compared with atmospheric controls held at 3 C, the total uptake at elevated pressure at 3 C was reduced, on an average, 55 times, CO2 respiration was reduced 45 times, and cellular assimilation was reduced 69 times. Rate of total uptake at elevated pressure was found to range from 4.0 X 10(-11) mug/cell per h for leucine to 2.61 X 10(-10) mug/cell per h for an amino acid mixture. Also, the percentage of total uptake at elevated pressures, respired as CO2, increased at the expense of cellular assimilation (ca. 22% increase). Two cellular-associated amino acid pools were detected, a large, loosely bound, outer pool and a small, tightly bound internal pool. The loosely bound outer pool was removed by a change in the pH of the incubation medium. Even though heterotrophic uptake and the outer, cellular-associated pool were markedly reduced at an elevated pressure, the percentage of total uptake calculated for the unincorporated, tightly bound, intracellular pool was 2 to 19 times that obtained for cultures held at 1 atm. The results were interpreted as indicating that bacterial metabolism and biosynthesis in the deep sea are markedly reduced, with a greater proportion of metabolic activity devoted to cellular maintenance.     

Collagenolytic Activity of Cured Hide Bacteria

S ummary : The aerobic and anaerobic collagenolytic activities of bacterial populations from 1 Australian and 2 South African cured hide samples at 0·85, 2·34, 7·0 and 10·0% of NaCl are described. The rate of denaturation of pure, undegraded collagen was determined by the release of amino acids and considerable collagenolysis was demonstrated. Collagenolytic activity was highest under aerobic conditions. Australian hides which produce uniformly good leather with little or no decay lacked collagenolytic bacteria.