Changes in diversity of cultured bacteria resistant to erythromycin and tetracycline in swine manure during simulated composting and lagoon storage.

UNLABELLED: This study investigated the impact of composting and lagoon storage on survival and change in diversity of tetracycline-resistant (Tc(r) ) and erythromycin-resistant (Em(r) ) bacteria and the resistance genes they carry in swine manure. Treatments were arranged as a 2 × 2 factorial design: composting vs lagoon storage and 0 vs 1% Surround WP Crop Protectant (a clay product) in three replicates. After 48 days of treatments, resistant bacteria were enumerated by selective plating and identified by 16S rRNA gene sequencing. The erm and the tet gene(s) carried by the resistant isolates were screened using class-specific PCR assays. The plate counts of Tc(r) and Em(r) bacteria decreased by 4-7 logs by composting, but only by 1-2 logs by the lagoon treatment. During the treatments, Acinetobacter gave way to Pseudomonas and Providencia as the largest resistant genera. The clay product had little effect on survival or diversity of resistant bacteria. Of six classes of erm and seven classes of tet genes tested, changes in prevalence were also noted. The results indicate that composting can dramatically shift Tc(r) and Em(r) bacterial populations, and composting can be an effective and practical approach to decrease dissemination of antibiotic resistance from swine farms to the environment. SIGNIFICANCE AND IMPACT OF THE STUDY: The presented research provided evidence that composting is much more effective than lagoon storage in dramatically decreasing culturable bacteria resistant to erythromycin and tetracycline in swine manure. Considerable diversity changes of resistant bacteria were also demonstrated during composting or lagoon storage. Overall, Acinetobacter was the major resistant genus in untreated swine manure, but pseudomonads and Providencia became the major resistant genera after the treatments. This is the first study that investigated diversity changes of cultured bacteria resistant to these two antibiotics during composting and lagoon storage of swine manure. New genes encoding resistance to the two antibiotics were also implied in the cultured isolates.

Bacterial community profiles on feathers during composting as determined by terminal restriction fragment length polymorphism analysis of 16S rDNA genes.

Composting is one of the more economical and environmentally safe methods of recycling feather waste generated by the poultry industry, since 90% of the feather weight consists of crude keratin protein, and feathers contain 15% N. However, the keratin in waste feathers is resistant to biodegradation and may require the addition of bacterial inocula to enhance the degradation process during composting. Two keratin-degrading bacteria isolated from plumage of wild songbirds and identified as Bacillus licheneformis (OWU 1411T) and Streptomyces sp. (OWU 1441) were inoculated into poultry feather composts (1.13 x 10(8) cfu g(-1) feathers) and co-composted with poultry litter and straw in 200-l compost vessels. Composting temperatures, as well as CO(2) and NH(3) evolution, were measured in these vessels to determine the effects of inoculation on the rate and extent of poultry feather decomposition during composting. Terminal restriction fragment length polymorphisms of 16S rRNA genes were used to follow changes in microbial community structure during composting. The results indicated that extensive carbon conversion occurred in both treatments (55.5 and 56.1%). The addition of the bacterial inocula did not enhance the rate of waste feather composting. The microbial community structure over time was very similar in inoculated and uninoculated waste feather composts.

Isolation and characterization of rhizobacteria from composts that suppress the severity of bacterial leaf spot of radish.

ABSTRACT Composts can induce systemic resistance in plants to disease. Unfortunately, the degree of resistance induced seems highly variable and the basis for this effect is not understood. In this work, only 1 of 79 potting mixes prepared with different batches of mature, stabilized composts produced from several different types of solid wastes suppressed the severity of bacterial leaf spot of radish caused by Xanthomonas campestris pv. armoraciae compared with disease on plants produced in a nonamended sphagnum peat mix. An additional batch of compost-amended mix that had been inoculated with Trichoderma hamatum 382 (T(382)), which is known to induce systemic resistance in plants, also suppressed the disease. A total of 11 out of 538 rhizobacterial strains isolated from roots of radish seedlings grown in these two compostamended mixes that suppressed bacterial leaf spot were able to significantly suppress the severity of this disease when used as inoculum in the compost-amended mixes. The most effective strains were identified as Bacillus sp. based on partial sequencing of 16S rDNA. These strains were significantly less effective in reducing the severity of this disease than T(382). A combined inoculum consisting of T(382) and the most effective rhizobacterial Bacillus strain was less effective than T(382) alone. A drench applied to the potting mix with the systemic acquired resistance-inducing chemical acibenzolar-S-methyl was significantly more effective than T(382) in several, but not all tests. We conclude that systemic suppression of foliar diseases induced by compost amendments is a rare phenomenon. Furthermore, inoculation of compost-amended potting mixes with biocontrol agents such as T(382) that induce systemic resistance in plants can significantly increase the frequency of systemic disease control obtained with natural compost amendments.

Evaluation of extraction and purification methods for obtaining PCR-amplifiable DNA from compost for microbial community analysis.

Analysis of microbial community structure in complex environmental samples using nucleic acid techniques requires efficient unbiased DNA extraction procedures; however, humic acids and other contaminants complicate the isolation of PCR-amplifiable DNA from compost and other organic-rich samples. In this study, combinations of DNA extraction and purification methods were compared based on DNA yield, humic acid contamination, PCR amplifiability, and microbial community structure assessed by terminal restriction fragment length polymorphisms (TRFLP) of amplified 16S rRNA genes. DNA yield and humic acid contamination, determined by A230, varied significantly between extraction methods. Humic acid contamination of DNA obtained from compost decreased with increasing salt concentration in the lysis buffer. DNA purified by gel permeation chromatography (Sepharose 4B columns) gave satisfactory PCR amplification with universal eubacterial 16S rRNA gene primers only when A260/A280 ratios exceeded 1.5. DNA purified with affinity chromatography (hydroxyapatite columns), and showing A260/A280 ratios as high as 1.8, did not show consistently satisfactory PCR amplification using the same 16S rRNA primers. Almost all DNA samples purified by agarose gel electrophoresis showed satisfactory PCR amplification. Principal components analysis (PCA) of TRFLP patterns differentiated compost types based on the presence/absence of peaks and on the height of the peaks, but differences in TRFLP patterns were not appreciable between extraction methods that yielded relatively pure DNA. High levels of humic acid contamination in extracted DNA resulted in TRFLP patterns that were not consistent and introduced a bias towards lower estimates of diversity.

Microbial degradation and humification of the lawn care pesticide 2,4-dichlorophenoxyacetic acid during the composting of yard trimmings.

The fate of the widely used lawn care herbicide 2,4-dichlorophenoxyacetic acid (2,4-D) during the composting of yard trimmings consisting of primarily leaves and grass is an important unexplored question. In this study, we determined the extent of 2,4-D mineralization, incorporation into humic matter, volatilization, and sorption during the composting of yard trimmings. Yard trimmings (2:1 [wt/wt] leaves-grass) were amended with 14C-ring-labeled 2,4-D (17 mg/kg of dry weight) and composted in a temperature-controlled laboratory scale compost system. During composting, thermophilic microbes were numerically dominant, reaching a maximum of 2 x 10(11)/g. At the end of composting, 46% of the organic matter (OM) present in the yard trimmings was lost and the compost was stable, with an oxygen uptake rate of 0.09 mg of O2 per g of OM per h, and was well humified (humification index, 0.39). Mineralization of the OM temporally paralleled mineralization of 2,4-D. In the final compost, 47% of the added 2,4-D carbon was mineralized, about 23% was complexed with high-molecular-weight humic acids, and about 20% was not extractable (humin fraction). Less than 1% of the added 14C was present in water expressed from the finished compost, suggesting a low potential for leaching of 2,4-D. Very little volatilization of 2,4-D occurred during composting. It is of interest that our results indicate active mineralization of 2,4-D at composting temperatures of 60 degrees C because microbial 2,4-D degradation at thermophilic temperatures has not been previously documented.

Determination of the respiration kinetics for mycelial pellets of Phanerochaete chrysosporium.

In mycelial pellet cultures of the white rot basidiomycete Phanerochaete chrysosporium, low oxygen concentration negatively affects the production of the extracellular lignin peroxidases and manganese peroxidases which are key components of the lignin-degrading system of this organism. To test the hypothesis that oxygen limitation in the pellets is responsible for this effect, oxygen microelectrodes were used to determine oxygen concentration gradients within the mycelial pellets of P. chrysosporium. Pellets were removed from oxygenated cultures, allowed to equilibrate with air, and probed with oxygen microelectrodes. The oxygen profiles were modelled assuming that O2 uptake follows a Michaelis-Menten relationship. The Vmax and Km values for oxygen uptake were 0.76 +/- 0.10 g/m3 of pellet per s and 0.5 +/- 0.3 g/m3, respectively. These kinetic values were used to predict respiration rates in air-flushed cultures, oxygen-flushed cultures, and cultures with large pellets (diameter greater than 6 mm). The predicted respiration rates were independently validated by experimentally measuring the evolution of carbon dioxide from whole cultures.

Role of manganese peroxidases and lignin peroxidases of Phanerochaete chrysosporium in the decolorization of kraft bleach plant effluent.

The role of lignin peroxidases (LIPs) and manganese peroxidases (MNPs) of Phanerochaete chrysosporium in decolorizing kraft bleach plant effluent (BPE) was investigated. Negligible BPE decolorization was exhibited by a per mutant, which lacks the ability to produce both the LIPs and the MNPs. Also, little decolorization was seen when the wild type was grown in high-nitrogen medium, in which the production of LIPs and MNPs is blocked. A lip mutant of P. chrysosporium, which produces MNPs but not LIPs, showed about 80% of the activity exhibited by the wild type, indicating that the MNPs play an important role in BPE decolorization. When P. chrysosporium was grown in a medium with 100 ppm of Mn(II), high levels of MNPs but no LIPs were produced, and this culture also exhibited high rates of BPE decolorization, lending further support to the idea that MNPs play a key role in BPE decolorization. When P. chrysosporium was grown in a medium with no Mn(II), high levels of LIPs but negligible levels of MNPs were produced and the rate and extent of BPE decolorization by such cultures were quite low, indicating that LIPs play a relatively minor role in BPE decolorization. Furthermore, high rates of BPE decolorization were seen on days 3 and 4 of incubation, when the cultures exhibit high levels of MNP activity but little or no LIP activity. These results indicate that MNPs play a relatively more important role than LIPs in BPE decolorization by P. chrysosporium.

When will a pulsar in supernova 1987a be seen?

The means by which a pulsar might be detected in the remnant of supernova 1987a in the Large Magelanic Cloud is examined. One possibility is that the slower-than-radioactive decay typically seen in the type II light curves is itself the sign of powering by the underlying pulsar, with the decline representing not the spinning down of the pulsar but rather the declining nebular opacity that would allow increasing amounts of the energy to escape as gamma rays. The test of this hypothesis (if the supernova conforms to type II expectations) would be to look for the "missing" energy in the form of those gamma rays that escape from the remnant instead of powering it.

Durability of the accretion disk of millisecond pulsars.

Pulsars with pulsation periods in the millisecond range are thought to be neutron stars that have acquired an extraordinarily short spin period through the accretion of stellar material spiraling down onto the neutron star from a nearby companion. Nearly all the angular momentum and most of the mass of the companion star is transferred to the neutron star. During this process, wherein the neutron star consumes its companion, it is required that a disk of stellar material be formed around the neutron star. In conventional models it is supposed that the disk is somehow lost when the accretion phase is finished, so that only the rapidly spinning neutron star remains. However, it is possible that, after the accretion phase, a residual disk remains in stable orbit around the neutron star. The end result of such an accretion process is an object that looks much like a miniature (about 100 kilometers), heavy version of Saturn: a central object (the neutron star) surrounded by a durable disk.

Ionosphere of venus: first observations of the dayside ion composition near dawn and dusk.

The first in situ measurements of the composition of the ionosphere of Venus are provided by independent Bennett radio-frequency ion mass spectrometers on the Pioneer Venus bits and orbiter spacecraft, exploring the dawn and duskside regions, respectively. An extensive composition of ion species, rich in oxygen, nitrogen, and carbon chemistry is idenitified. The dominant topside ion is O(+), with C(+), N(+), H(+), and He(+) as prominent secondary ions. In the lower ionosphere, the ionzization peak or F(1) layer near 150 kilometers reaches a concentration of about 5 x l0(3) ions per cubic centimeter, and is composed of the dominant molecular ion, O(2)(+), with NO(+), CO(+), and CO(2)(+), constituting less than 10 percent of the total. Below the O(+) peak near 200 kilometers, the ions exhibit scale heights consistent with a neutral gas temperature of about 180 K near the terminator. In the upper ionosphere, scale heights of all species reflect the effects of plasma transport, which lifts the composition upward to the often abrupt ionopause, or thermal ion boundary, which is observed to vary in height between 250 to 1800 kilometers, in response to solar wind dynamics.

Ionosphere of venus: first observations of the effects of dynamics on the dayside ion composition.

Bennett radio-frequency ion mass spectrometers have returned the first in situ measurements of the Venus dayside ion composition, including evidence of pronounced structural variability resulting from a dynamic interaction with the solar wind. The ionospheric envelope, dominated above 200 kilometers by O(+), responds dramatically to variations in the solar wind pressure, Which is observed to compress the thermal ion distributions from heights as great as 1800 kilometers inward to 280 kilometers. At the thermal ion boundary, or ionopause, the ambient ions are swept away by the solar wind, such that a zone of accelerated suprathermnal plasma is encountered. At higher altitudes, extending outward on some orbits for thousands of kilometers to the bows shock, energetic ion currents are detected, apparently originating from the shocked solar wind plasma. Within the ionosphere, observations of pass-to-pass differences in the ion scale heights are indicative of the effects of ion convection stimlulated by the solar wind interaction.

Lunar atmosphere as a source of argon-40 and other lunar surface elements.

The lunar atmosphere is the likely source of excess argon-40 in lunar surface material; about 8.5 percent of the argon-40 released into the lunar atmosphere will be implanted in the surface material by photoionization and subsequent interaction with fields in the solar wind. The atmosphere is also likely to be the source of other unexpected surface elements or of solar wind elements that impact from non-solar wind directions.