Ventilation rates in large commercial layer hen houses with two-year continuous monitoring.

1. Ventilation controls the indoor environment and is critical for poultry production and welfare. Ventilation is also crucial for assessing aerial pollutant emissions from the poultry industry. Published ventilation data for commercial layer houses have been limited, and are mostly based on short-term studies, mainly because monitoring airflow from large numbers of fans is technically challenging. 2. A two-year continuous ventilation monitoring trial was conducted at two commercial manure belt houses (A and B), each with 250 000 layers and 88 130-cm exhaust fans. All the fans were individually monitored with fan rotational speed sensors or vibration sensors. Differential static pressures across the house walls were also measured. Three fan performance assessment methods were applied periodically to determine fan degradations. Fan models were developed to calculate house ventilations. 3. A total of 693 and 678 complete data days, each containing >16 h of valid ventilation data, were obtained in houses A and B, respectively. The two-year mean ventilation rates of houses A and B were 2·08 and 2·10 m(3) h(-1) hen(-1), corresponding to static pressures of -36·5 and -48·9 Pa, respectively. For monthly mean ventilation, the maximum rates were 4·87 and 5·01 m(3) h(-1) hen(-1) in July 2008, and the minimum were 0·59 and 0·81 m(3) h(-1) hen(-1) in February 2008, for houses A and B, respectively. 4. The two-year mean ventilation rates were similar to those from a survey in Germany and a 6-month study in Indiana, USA, but were much lower than the 8·4 and 6·2 m(3) h(-1) hen(-1) from a study in Italy. The minimum monthly mean ventilation rates were similar to the data obtained in winter in Canada, but were lower than the minimum ventilation suggested in the literature. The lower static pressure in house B required more ventilation energy input. The two houses, although identical, demonstrated differences in indoor environment controls that represented potential to increase ventilation energy efficiency, and reduce carbon footprints and operational costs.

Degradation of straight-chain aliphatic and high-molecular-weight polycyclic aromatic hydrocarbons by a strain of Mycobacterium austroafricanum.

AIMS: Our goal was to characterize a newly isolated strain of Mycobacterium austroafricanum, obtained from manufactured gas plant (MGP) site soil and designated GTI-23, with respect to its ability to degrade polycyclic aromatic hydrocarbons (PAHs). METHODS AND RESULTS: GTI-23 is capable of growth on phenanthrene, fluoranthene, or pyrene as a sole source of carbon and energy; it also extensively mineralizes the latter two in liquid culture and is capable of extensive degradation of fluorene and benzo[a]pyrene, although this does not lead in either of these cases to mineralization. Supplementation of benzo[a]pyrene-containing cultures with phenanthrene had no significant effect on benzo[a]pyrene degradation; however, this process was substantially inhibited by the addition of pyrene. Extensive and rapid mineralization of pyrene by GTI-23 was also observed in pyrene-amended soil. CONCLUSIONS: Strain GTI-23 shows considerable ability to mineralize a range of polycyclic aromatic hydrocarbons, both in liquid and soil environments. In this regard, GTI-23 differs markedly from the type strain of Myco. austroafricanum (ATCC 33464); the latter isolate displayed no (or very limited) mineralization of any tested PAH (phenanthrene, fluoranthene or pyrene). When grown in liquid culture, GTI-23 was also found to be capable of growing on and mineralizing two aliphatic hydrocarbons (dodecane and hexadecane). SIGNIFICANCE AND IMPACT OF THE STUDY: These findings indicate that this isolate of Myco. austroafricanum may be useful for bioremediation of soils contaminated with complex mixtures of aromatic and aliphatic hydrocarbons.

Effects of alkylphosphates and nitrous oxide on microbial degradation of polycyclic aromatic hydrocarbons.

We conducted a series of liquid-culture experiments to begin to evaluate the abilities of gaseous sources of nitrogen and phosphorus to support biodegradation of polycyclic aromatic hydrocarbons (PAHs). Nutrients examined included nitrous oxide, as well as triethylphosphate (TEP) and tributylphosphate (TBP). Cultures were established using the indigenous microbial populations from one manufactured gas plant (MGP) site and one crude oil-contaminated drilling field site. Mineralization of phenanthrene was measured under alternative nutrient regimes and was compared to that seen with ammoniacal nitrogen and PO(4). Parallel cultures were used to assess removal of a suite of three- to five-ring PAHs. In summary, the abilities of the different communities to degrade PAH when supplemented with N(2)O, TEP, and TBP were highly variable. For example, in the MGP soil, organic P sources, especially TBP, supported a considerably higher degree of removal of low-molecular-weight PAHs than did PO(4); however, loss of high-molecular-weight compounds was impaired under these conditions. The disappearance of most PAHs was significantly less in the oil field soil when organophosphates were used. These results indicate that the utility of gaseous nutrients for PAH bioremediation in situ may be limited and will very likely have to be assessed on a case-by-case basis.

Expression of lip genes during growth in soil and oxidation of anthracene by Phanerochaete chrysosporium.

mRNA extraction from soil and quantitation by competitive reverse transcription-PCR were combined to study the expression of the 10 known lignin peroxidase (lip) genes in anthracene-transforming soil cultures of Phanerochaete chrysosporium. Levels of extractable lipA transcript and protein (LiP H8) were well correlated, although they were separated by a 2-day lag period. The patterns of transcript abundance over time in soil-grown P. chrysosporium varied among the nine lip mRNAs detected; comparison with lip gene expression under different liquid culture conditions suggested an early phase of carbon limitation for the cultures as a whole, which was followed by a transition to nitrogen starvation. Anthracene transformation occurred throughout the 25-day course of the experiment and, therefore, likely involves mechanisms distinct from those involved in oxidation of non-LiP substrate polycyclic aromatic hydrocarbons.

Manganese peroxidase mRNA and enzyme activity levels during bioremediation of polycyclic aromatic hydrocarbon-contaminated soil with Phanerochaete chrysosporium.

mRNA extraction from soil and quantitation by competitive reverse transcription-PCR were combined to study the expression of three manganese peroxidase (MnP) genes during removal of polycyclic aromatic hydrocarbons from cultures of Phanerochaete chrysosporium grown in presterilized soil. Periods of high mnp transcript levels and extractable MnP enzyme activity were temporally correlated, although separated by a short (1- to 2-day) lag period. This time frame also coincided with maximal rates of fluorene oxidation and chrysene disappearance in soil cultures, supporting the hypothesis that high ionization potential polycyclic aromatic hydrocarbons are oxidized in soil via MnP-dependent mechanisms. The patterns of transcript abundance over time in soil-grown P. chrysosporium were similar for all three of the mnp mRNAs studied, indicating that transcription of this gene family may be coordinately regulated under these growth conditions.

Fluorene Oxidation In Vivo by Phanerochaete chrysosporium and In Vitro during Manganese Peroxidase-Dependent Lipid Peroxidation.

The oxidation of fluorene, a polycyclic hydrocarbon which is not a substrate for fungal lignin peroxidase, was studied in liquid cultures of Phanerochaete chrysosporium and in vitro with P. chrysosporium extracellular enzymes. Intact fungal cultures metabolized fluorene to 9-hydroxyfluorene via 9-fluorenone. Some conversion to more-polar products was also observed. Oxidation of fluorene to 9-fluorenone was also obtained in vitro in a system that contained manganese(II), unsaturated fatty acid, and either crude P. chrysosporium peroxidases or purified recombinant manganese peroxidase. The oxidation of fluorene in vitro was inhibited by the free-radical scavenger butylated hydroxytoluene but not by the lignin peroxidase inhibitor NaVO(inf3). Manganese(III)-malonic acid complexes could not oxidize fluorene. These results indicate that fluorene oxidation in vitro was a consequence of lipid peroxidation mediated by P. chrysosporium manganese peroxidase. The rates of fluorene and diphenylmethane disappearance in vitro were significantly faster than those of true polycyclic aromatic hydrocarbons or fluoranthenes, whose rates of disappearance were ionization potential dependent. This result indicates that the initial oxidation of fluorene proceeds by mechanisms other than electron abstraction and that benzylic hydrogen abstraction is probably the route for oxidation.

Polycyclic aromatic hydrocarbon-degrading capabilities of Phanerochaete laevis HHB-1625 and its extracellular ligninolytic enzymes.

The ability of Phanerochaete laevis HHB-1625 to transform polycyclic aromatic hydrocarbons (PAHs) in liquid culture was studied in relation to its complement of extracellular ligninolytic enzymes. In nitrogen-limited liquid medium, P. laevis produced high levels of manganese peroxidase (MnP). MnP activity was strongly regulated by the amount of Mn2+ in the culture medium, as has been previously shown for several other white rot species. Low levels of laccase were also detected. No lignin peroxidase (LiP) was found in the culture medium, either by spectrophotometric assay or by Western blotting (immunoblotting). Despite the apparent reliance of the strain primarily on MnP, liquid cultures of P. laevis were capable of extensive transformation of anthracene, phenanthrene, benz[a]anthracene, and benzo[a]pyrene. Crude extracellular peroxidases from P. laevis transformed all of the above PAHs, either in MnP-Mn2+ reactions or in MnP-based lipid peroxidation systems. In contrast to previously published studies with Phanerochaete chrysosporium, metabolism of each of the four PAHs yielded predominantly polar products, with no significant accumulation of quinones. Further studies with benz[a]anthracene and its 7,12-dione indicated that only small amounts of quinone products were ever present in P. laevis cultures and that quinone intermediates of PAH metabolism were degraded faster and more extensively by P. laevis than by P. chrysosporium.

One-electron oxidation in the degradation of creosote polycyclic aromatic hydrocarbons by Phanerochaete chrysosporium.

The abilities of whole cultures of Phanerochaete chrysosporium and P. chrysosporium manganese peroxidase-mediated lipid peroxidation reactions to degrade the polycyclic aromatic hydrocarbons (PAHs) found in creosote were studied. The disappearance of 12 three- to six-ring PAHs occurred in both systems. Both in vivo and in vitro, the disappearance of all PAHs was found to be very strongly correlated with ionization potential. This was true even for compounds beyond the ionization potential thresholds of lignin peroxidase and Mn3+. Deviations from this correlation were seen in the cases of PAHs which are susceptible to radical addition reactions. These results thus begin to clarify the mechanisms of non-lignin peroxidase-labile PAH degradation in the manganese peroxidase-lipid peroxidation system and provide further evidence for the ability of this system to explain the in vivo oxidation of these compounds.