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.

Bacterial inoculum enhances keratin degradation and biofilm formation in poultry compost.

Native microbial populations can degrade poultry waste, but the process can be hastened by using feather-degrading bacteria. Strains of Bacillus licheniformis and a Streptomyces sp. isolated from the plumage of wild birds were grown in a liquid basal medium and used to inoculate feathers in compost bioreaction vessels. Control vessels had only basal medium added to the feathers, litter and straw. Temperature, ammonia, carbon and nitrogen were monitored for 4 weeks. Scanning electron microscopy of the feather samples showed more complete keratin-degradation, more structural damage, and earlier microbial biofilm formation on inoculated feathers than on uninoculated feathers. A diverse community of aerobic bacteria and fungi were cultured early, but declined rapidly. Thermophilic B. licheniformis and Streptomyces spp. were abundant throughout. Enteric gram-negative bacteria, (e.g., Salmonella, E. coli) originally found on waste feathers were not recovered after day 4. Vessel temperatures reached 64-71 degrees C within 36 h and stabilized at 50 degrees C. When tumble-mixed at day 14, renewed activity peaked at 59 degrees C and quickly dropped as available carbon was used. Feathers soaked in an inoculum of B. licheniformis and Streptomyces degraded more quickly and more completely than feathers that were not presoaked. Inoculation of feather waste could improve composting of the large volume of feather waste generated every year by poultry farms and processing plants.

Long-term correction of canine hemophilia B by gene transfer of blood coagulation factor IX mediated by adeno-associated viral vector.

Hemophilia B is a severe X-linked bleeding diathesis caused by the absence of functional blood coagulation factor IX, and is an excellent candidate for treatment of a genetic disease by gene therapy. Using an adeno-associated viral vector, we demonstrate sustained expression (>17 months) of factor IX in a large-animal model at levels that would have a therapeutic effect in humans (up to 70 ng/ml, adequate to achieve phenotypic correction, in an animal injected with 8.5x10(12) vector particles/kg). The five hemophilia B dogs treated showed stable, vector dose-dependent partial correction of the whole blood clotting time and, at higher doses, of the activated partial thromboplastin time. In contrast to other viral gene delivery systems, this minimally invasive procedure, consisting of a series of percutaneous intramuscular injections at a single timepoint, was not associated with local or systemic toxicity. Efficient gene transfer to muscle was shown by immunofluorescence staining and DNA analysis of biopsied tissue. Immune responses against factor IX were either absent or transient. These data provide strong support for the feasibility of the approach for therapy of human subjects.

Using activity-based management to control costs & achieve organization goals.

Activity-based management (ABM) is a management process that focuses on improving costs and outcomes. It derives useful information based on the way people think (their activities) rather than traditional expense categories. ABM supports outcomes, quality, teams, re-engineering, empowerment, and continuous improvement. It is a process that providers may want to adopt in light of new Medicare reimbursement practices.

Characterization of the mutant N-acetylglucosaminylphosphotransferase in I-cell disease and pseudo-Hurler polydystrophy: complementation analysis and kinetic studies.

Complementation was examined among various types of I-cell disease and pseudo-Hurler polydystrophy by monitoring N-acetylglucosaminylphosphotransferase activity in multinucleated cells produced by fusing pair combinations of cultured skin fibroblasts. Patients with the classical forms of these disorders (5 I-cell disease and 3 pseudo-Hurler polydystrophy cell lines) comprised one complementation group and 5 cell lines from patients with variant forms of pseudo-Hurler polydystrophy comprised a distinct complementation group. In the first group, total or partial deficiency of the transferase activity was demonstrated with both natural (lysosomal enzymes) and artificial (alpha-methylmannoside) acceptor substrates with low Vmax but apparently normal Km values for the donor (UDP-GlcNAc) and acceptor (alpha-methylmannoside) substrates. The activity toward artificial substrate could be inhibited by adding exogenous lysosomal enzyme preparations to the reaction mixture. In the second group, the cells demonstrated deficiency of the transferase activity toward lysosomal enzyme acceptors but had normal activity toward alpha-methylmannoside acceptor and this activity could not be inhibited by the addition of exogenous lysosomal enzyme preparations. These findings suggest that N-acetylglucosaminylphosphotransferase is composed of at least two distinct subunits, a catalytic subunit which is absent or defective in the first complementation group, and a recognition subunit which is altered or deficient in the second group.