RESUMO
A composting-accelerating bio-inoculant (Bacillus subtilis, Talaromyces sayulitensis (HC1), Steinernema sp., and Heterorhabditis sp.) was evaluated in a composting process made up of a different mix of wood chips, pig manure, urine, and swine mortality (raw material RM). Three different treatments (T1, T2, and T3) were assessed, and physicochemical, microbiological, and entomological evaluations were carried out at 0 and 45 days of the composting process. The highest organic nitrogen (1.34 %) concentration was detected in swine mortality, whereas the highest total oxidizable organic carbon (39.1 %) concentration was observed in wood chips. Salmonella spp., was not identified in any of the raw materials. Clostridium spp., count was 5.5, 2.0, and 1.0 Log10 unit, for pig manure, wood chips, and swine mortality, respectively. Pig manure, swine mortality, and wood chip total coliform count was 6.21, 5.32, and 1 Log10 unit, respectively. Helminth eggs were not detected in any of the RM and Cryptosporidium spp., oocysts were occasionally found in pig manure and wood chips. Several types of flies were identified, Musca domestica, Muscina stabulans, Stomoxys calcitrans, Fannia canicularis, Sarcophaga sp., and Calliphora sp. Treatment 3 (45.11 % swine mortality, 33.33 % wood chips, and 21.55 %, urine and bio-inoculant) had the greatest total oxidizable organic carbon availability, the highest carbon/nitrogen (C/N) ratio (20.67, p < 0.05), and the lowest dipterous larvae count. Moreover, Salmonella sp., was not observed and had only low Clostridium spp., and fecal coliform count. The bio-inoculant's effect on C/N ratio, cation exchange capacity, and electrical conductivity were beneficial, and resulted in production of a fertilizer complying with EPA 600/1-87-014, EPA 40 CFR Part 258, and NTC5167/11 norms. According to the characterization protocols used in this study the compost was apparently free from bacterial and parasitic pathogens and minimal dipteran counts. Last, maturation time was 15 days shorter compared with control (C4).
RESUMO
Laccases (E.C. 1.10.3.2) are multicopper oxidases of great importance in the industry due to their non-specificity and high oxidative potential. Laccases are useful to bleach synthetic dyes, oxidize phenolic compounds and degrade pesticides, among others. Hence, the objective of this work was to optimize low cost culture media for recombinant (rPOXA 1B) laccase production from Pleurotus ostreatus in Pichia pastoris. To this end, low cost nitrogen sources were studied, such as malt extract, isolated soy protein and milk serum. Following, two central composite designs (CCD) were performed. In CCD-1 different concentrations of glucose USP (0-13.35 gL-1), protein isolated soy protein (5-25 gL-1), malt extract (3.5-17.5 gL-1) and (NH4)2SO4 (1.3-6.5 gL-1) were evaluated. In CCD-2 only different concentrations of glucose USP (7.9-22 gL-1) and isolated soy protein (15.9-44.9 gL-1) were evaluated. CCD-2 results led to a One Factor Experimental design (OFED) to evaluate higher isolated soy protein (20-80 gL-1) concentrations. In all designs, (CCD-1, CCD-2 and OFED) CuSO4 (0.16 gL-1) and chloramphenicol (0.1 gL-1) concentrations remained unchanged. For the OFED after sequential statistical optimization, an enzyme activity of 12,877.3 ± 481.2 UL-1 at 168 h was observed. rPOXA 1B activity increased 30.54 % in comparison with CCD-2 results. Final composition of optimized media was: 20 gL-1 glucose USP, 50 gL-1 isolated soy protein 90 % (w/w), 11.74 gL-1 malt extract, and 4.91 gL-1 (NH4)2SO4. With this culture media, it was possible to reduce culture media costs by 89.84 % in comparison with improved culture media previously described by our group.