Effect of Broiler Litter Based Complete Feed Fermentation Time on Nutrient Content and In vitro Digestibility

Broiler litter waste is increasing as the population of broiler chickens increases, on the other hand the need for ruminant feed is increasing so that alternative feeds are needed in the form of complete feed made from litter. This study was aimed to determine the effect of fermentation time on the nutritional content and digestibility of the complete feed. A completely randomized design with 4 treatments and 5 replications were applied in this study. Treatment T0 = not fermented; T1 = fermentation for 10 days; T2 = fermentation for 24 days; T3 = fermentation for 38 days. Parameters observed were nutrient content, VFA, NH3, dry matter digestibility and organic matter digestibility in vitro using cow rumen. Results showed that  fermention of complete feed with 2.5% EM4 starter and 5% probiofeed for 38 days (T3) had a significant effect (P0.05) on nutrient content, dry matter digestibility, organic matter digestibility, VFA and NH3.  The T3 significantly affected ash content, crude fat, crude protein, BETN, TDN, dry matter digestibility, organic matter digestibility, VFA, and NH3 but had no significant effect on water content and crude fiber. It is concluded that the complete feed could be used as an alternative feed for ruminants, and it is easy to obtain, cheap and able to reduce environmental pollution

Molecular Cloning and Expression Analysis of the Endogenous Cellulase Gene MaCel1 in Monochamus alternatus

The purpose of this study was to characterize the endogenous cellulase gene MaCel1 of Monochamus alternatus, which is an important vector of Bursaphelenchus xylophilus, a pine wood nematode, which causes pine wilt disease (PWD). In this study, MaCel1 was cloned by rapid amplification of cDNA end (RACE), and its expression analyzed by RT-qPCR (real-time quantitative PCR detecting). A total of 1778 bp of cDNA was obtained. The encoding region of this gene was 1509 bp in length, encoding a protein containing 502 amino acids with a molecular weight of 58.66 kDa, and the isoelectric point of 5.46. Sequence similarity analysis showed that the amino acids sequence of MaCel1 had high similarity with the beta-Glucosinolate of Anoplophora glabripennis and slightly lower similarity with other insect cellulase genes (GH1). The beta-D-Glucosidase activity of MaCel1 was 256.02 +/- 43.14 U/L with no beta-Glucosinolate activity. MaCel1 gene was widely expressed in the intestine of M. alternatus. The expression level of MaCel1 gene in male (3.46) and female (3.51) adults was significantly higher than that in other developmental stages, and the lowest was in pupal stage (0.15). The results will help reveal the digestive mechanism of M. alternatus and lay the foundation for controlling PWD by controlling M. alternatus

Metatranscriptomic profiles of Eastern subterranean termites, Reticulitermes flavipes (Kollar) fed on second generation feedstocks

Background: Second generation lignocellulosic feedstocks are being considered as an alternative to first generation biofuels that are derived from grain starches and sugars. However, the current pre-treatment methods for second generation biofuel production are inefficient and expensive due to the recalcitrant nature of lignocellulose. In this study, we used the lower termite Reticulitermes flavipes (Kollar), as a model to identify potential pretreatment genes/enzymes specifically adapted for use against agricultural feedstocks. Results: Metatranscriptomic profiling was performed on worker termite guts after feeding on corn stover (CS), soybean residue (SR), or 98% pure cellulose (paper) to identify (i) microbial community, (ii) pathway level and (iii) gene-level responses. Microbial community profiles after CS and SR feeding were different from the paper feeding profile, and protist symbiont abundance decreased significantly in termites feeding on SR and CS relative to paper. Functional profiles after CS feeding were similar to paper and SR; whereas paper and SR showed different profiles. Amino acid and carbohydrate metabolism pathways were downregulated in termites feeding on SR relative to paper and CS. Gene expression analyses showed more significant down regulation of genes after SR feeding relative to paper and CS. Stereotypical lignocellulase genes/enzymes were not differentially expressed, but rather were among the most abundant/constitutively-expressed genes. Conclusions: These results suggest that the effect of CS and SR feeding on termite gut lignocellulase composition is minimal and thus, the most abundantly expressed enzymes appear to encode the best candidate catalysts for use in saccharification of these and related second-generation feedstocks. Further, based on these findings we hypothesize that the most abundantly expressed lignocellulases, rather than those that are differentially expressed have the best potential as pretreatment enzymes for CS and SR feedstocks. © 2015 Rajarapu et al

Biosugar production from oil palm mesocarp fiber using crude lignocellulosic degrading enzymes from rhizomucor pusillus AK2

The oil palm industry is undergoing global expansion as a result of the demand of oil palm. Consequently, similar expansion is occurring in amounts of lignocellulosic residues generated by the industries. The conversion of these residues to biosugar is limited by the uneconomical hydrolysis process and the high cost of lignocellulolytic enzymes. Thus, this study aimed at the production of enzymes for efficient degradation of oil palm mesocarp fiber (OPMF). The sum of 6 fungi isolates, AK1 to AK6, were isolated from OPMF, and screened for lignocellulolytic enzymes production on selective medium. Further screening of these isolates were carried out on untreated OPMF using solid state fermentation (SSF). Design-Expert software version 7.0, was used to operate the 2 level fractional factorial design and Central Composite Design (CCD) for the screening and optimization of significant factors affecting lignocelluose degrading enzymes production. The activity of crude enzyme, Viscozyme and Celluclast were evaluated based on the generation of biosugar from OPMF by determining the effects of pretreatments (2% (v/v) HNO3, 2% (w/v) NaOH and 2% (w/v) 1-Butyl-3-methylimidazolium chloride), solid loading (1-4 % w/v) and enzyme cocktail (1:1 (v/v), crude enzyme and Viscozyme, crude enzyme and celluclast, viscozyme and celluclast, and 1:1:1 (v/v) crude enzyme, Viscozyme and Celluclast). Isolate AK2 exhibited potential for lignocellulase enzyme production based on the hydrolysis zones >1.5 mm on selective media and producing CMCase (25.4 U/g), FPase, (5.5 U/g), β-glucosidase (9.8 U/g), Xylanase (68.4 U/g) and MnP (4.9 U/g) at exceptional level. The isolate was thus identified by 18S RNA gene sequencing using a universal primer ITS1-F and ITS4-R as Rhizomucor pusillus AK2 (KY583064). From the 2 level fractional factorial design pH, temperature, inoculum size and moisture content are the significant factors affecting the production of lignocellulolytic enzymes. Xylanase was observed to be the highest activity in the lignocellulolytic enzymes cocktail (111.01 U/g). Therefore, CCD was carried out focussing on xylanase production. At the optimum condition, xylanase (128 U/g), was obtained at pH 4.98, temperature 40.27 °C, inoculum size at 108.2 spores/g and moisture at 80.64% using CCD. The regression model of the ANOVA was found to be significant with p<0.0001 and R2 of 0.9831.Biochemical characterization of the crude enzymes indicated that the enzyme was stable at pH of 4 to 6 and temperatures of 30 to 60 °C. Enzymatic saccharification was carried out with the crude enzymes comparative to Viscozyme and Celluclast. Maximum sugar production was obtained from celluclast-saccharified OPMF (1.2 g/L) using 1% (w/v) NaOH pretreated OPMF. Maximum reducing sugar generated from enzyme cocktail was 1.8 g/L obtained from 1% (w/v) NaOH pretreated OPMF which translates to polyoses; glucose (2.59 g/L), xylose (2.1 g/L) and arabinose (0.254 g/L) at 1:1:1 (v/v) crude enzyme, viscozyme and celluclast. The performance of the enzyme cocktail customized in this study is superior to that of the individual and cocktail of commercial enzyme. The study also indicates the potential of OPMF as both a substrate for biosugar and lignocellulase enzyme production from R. pusillus AK2

Comparison of various culture media effectiveness in the isolation of bacteria from Pekan peat swamp forest soil

Aims: Previously described as non-favorable-microbial habitat, peat swamp forest has its own features, which are extremely acidic, poor in nutrient, water-logged and anoxic environment where rate of decomposition of plant litters is quiet slow. Interestingly, current research has proven that there is diversity of microbial communities in this ecosystem. The main objective of this study is to isolate bacteria from Pekan peat swamp forest soil that play a role in the decomposition of plant litters through cultivation on different agar-based medium. The success of isolation of bacteria from this neglected habitat could open the opportunity in unleashing the specific role of bacteria in peat swamp plant litter degradation as well as potential biotechnological application of these bacteria in lignocellulose-related industry. Methodology and results: To mimic the peat condition that is low in nutrient and comprised of plant debris, M1 and peat agar supplemented with cellulose, glucose, lignin and xylan were used. Specifically, for the isolation of actinomycetes, dry and wet heat pre-treatments were applied to the soil samples. Then, the samples were cultivated on three different agars which were oatmeal agar as well as M1 and peat agar supplemented with glucose. Enrichment method was applied in the isolation of cellulase-producing bacteria. It was found that higher number of bacteria and actinomycetes were successfully isolated from peat agar, followed by oatmeal agar and M1. In fact, more actinomycetes were isolated from soil that was treated with wet heat pre-treatment compared to dry heat pre-treatment and on peat agar compared to M1 and oatmeal agar. This finding is promising, indicating that the application of peat water in the agar-based medium is useful to mimic the actual environment of peat swamp and increase the possibility to isolate indigenous bacteria. Primary screening of isolates from samples enriched with carboxymethyl cellulose (CMC) showed positive result of decolourisation zone on Azo-CM-Cellulose agar indicating the ability of isolates to degrade cellulose compound. Conclusions, significance and impacts of study: The study indicates the effectiveness of different culture media in successful isolation of bacteria including actinomycetes. Using the enrichment method, bacteria that are able to degrade cellulose compound was successfully isolated even though it is well known that plant litter degradation in the peat swamp environment happens at very slow rates

Potensi Lignoselulolitik Dan Selulolitik Baglog Bekas Jamur Tiram Putih (Pleurotus osteratus L.) Sebagai Biang Kompos Dalam Penurunan Bobot Sampah Organik

Penelitian Potensi Lignoselulolitik Dan Selulolitik Baglog Bekas Jamur Tiram Putih (Pleurotus osteratus L.) Sebagai Biang Kompos Dalam Penurunan Bobot Sampah Organik dilaksanakan di Laboratorium Riset Mikrobiologi, Fisiologi Tumbuhan, Jurusan Biologi, Fakultas Matematika Dan Ilmu Pengetahuan Alam, dan di Laboratorium Kimia Tanah, Jurusan Ilmu Tanah, Fakultas Pertanian, Unversitas Andalas, Padang. Penelitian ini bertujuan Untuk Menentukan keberadaan enzim dan aktivitas enzim (Selulase, Lignoselulase) baglog bekas jamur tiram putih (Pleurotus osteratus L.), Mengetahui potensi lignoselulolitik dan selulolitik baglog bekas jamur tiram putih (Pleurotus osteratus L.) sebagai biang dalam pengomposan terhadap penurunan bobot sampah organik. Metode penelitian yang digunakan adalah metode survey, dianalisis secara deskriptif dan disajikan dalam bentuk grafik, tabel dan gambar. Hasil penelitian menunjukkan bahwa di dalam baglog bekas jamur tiram putih (Pleurotus osteratus L.) nilai rata-rata aktivitas enzim selulase yang di peroleh sebesar 0,129 μmol/g. Sedangkan nilai rata-rata aktivitas lignoselulase sebesar 1.073,172 U/g. Potensi Lignoselulolitik dan selulolitik dalam baglog bekas jamur tiram putih (Pleurotus osteratus L.) dapat di jadikan biang dalam penurunan bobot sampah sebesar 39%, dan nilai C/N sebesar 16,222