Use of Rhodopseudomonas palustris P1 stimulated growth by fermented pineapple extract to treat latex rubber sheet wastewater to obtain single cell protein

Latex rubber sheet wastewater (non sterile wastewater: RAW) was treated efficiently using a stimulated Rhodopseudomonas palustris P1 inoculum with added fermented pineapple extract (FPE) under microaerobic light conditions. Optimization of wastewater treatment conditions using a central composite design (CCD) found that a 3 % stimulated P1 inoculum with 0.9 % added FPE and a 4-day retention time (RT) were the most suitable conditions. Calculations from CCD experiments predicted that a chemical oxygen demand (COD) of 3,005 mg/L could be 98 % removed, together with 79 % of suspended solids (SS) and 72 % of total sulfide (TtS). No H₂S was detected, production costs were low and single cell protein (SCP) was a by-product. The results of the verification test had an error of only 4–8 % and confirmed removal of COD (initial COD 2,742 mg/L), SS and TtS at 94 %, 75 % and 66 %, respectively. These values were less than the best set obtained from the CCD experiment (2 % stimulated P1 inoculum, 0.75 % FPE and 4 days RT); upon repeating, this set could reduce 96 % of the COD, 78 % SS and 71 % TtS. The treated wastewater met the standard guidelines for irrigation use and no H₂S was detected. The biomass obtaining after wastewater treatment from the best set consisted mostly of R. palustris P1; the biomass of this set had 65 % protein, 3 % fat, 8 % carbohydrate, 14 % ash and 10 % moisture. The results demonstrated that an inoculum of stimulated P1 grew well in RAW supplemented with FPE and could be considered to be an appropriate technology for effectively treating wastewater, with SCP as a by-product.     

Rapid Synthesis of a Long Double-Stranded Oligonucleotide from a Single-Stranded Nucleotide Using Magnetic Beads and an Oligo Library

Chemical synthesis of oligonucleotides is a widely used tool in the field of biochemistry. Several methods for gene synthesis have been introduced in the growing area of genomics. In this paper, a novel method of constructing dsDNA is proposed. Short (28-mer) oligo fragments from a library were assembled through successive annealing and ligation processes, followed by PCR. First, two oligo fragments annealed to form a dsDNA molecule. The double-stranded oligo was immobilized onto magnetic beads (solid support) via streptavidin-biotin binding. Next, single-stranded oligo fragments were added successively through ligation to form the complete DNA molecule. The synthesized DNA was amplified through PCR and gel electrophoresis was used to characterize the product. Sanger sequencing showed that more than 97% of the nucleotides matched the expected sequence. Extending the length of the DNA molecule by adding single-stranded oligonucleotides from a basis set (library) via ligation enables a more convenient and rapid mechanism for the design and synthesis of oligonucleotides on the go. Coupled with an automated dispensing system and libraries of short oligo fragments, this novel DNA synthesis method would offer an efficient and cost-effective method for producing dsDNA.     

Histological structure of the digestive tract,liver, and pancreas of Ambassis nalua (Hamilton, 1822) with ultrastructural details of the gastric gland

The scalloped perchlet Ambassis nalua is one of the dominant fishes in the Estuarine Pranburi River, Thailand. It is suggested that this fish is in the secondary trophic level with a carnivorous nature. Studies on digestive system will help us further identify the niche of this species in the food web/food chain. The present study therefore aimed to report the detailed structure and ultrastructure of A. nalua digestive system. Fish samples (n = 30) with a total length of 5.7 ± 0.5 cm were obtained using beach seines from the Estuarine Pranburi River. Their digestive tract length and intestine coeficient were 3.6 ± 0.07 cm and 0.91, respectively. Light microscopic observation showed that the digestive wall comprised four layers, namely mucosa, submucosa, muscularis, and serosa. The prominent mucous-secreting cells were found in the mucosal oesophagus. The stomach had many gastric folds, with height and width being 649.76 ± 85.15 and 370.30 ± 68.56 μm, respectively. Gastric glands were found in the anterior stomach but not in the posterior stomach. Each gastric gland was made up of a single type of columnar cells. The gastric cells were ultrastructurally characterized by numerous mitochondria and well-developed secretory granules of varying sizes. A few small vacuoles were also identified in the apical area of the gastric cells. The intestine had two regions (anterior and posterior intestines), and pyloric caecum was absent. The density of the goblet cell was significantly higher in the posterior intestine. These results provide basic knowledge of the digestive system of A. nalua, and the low intestine coefficient and the absence of pyloric caecum suggest the carnivorous feeding habit of this species.     

Enhancing digestion efficiency of POME in anaerobic sequencing batch reactor with ozonation pretreatment and cycle time reduction

Ozonation pretreatment was applied to palm oil mill effluent (POME) prior to anaerobic digestion using the anaerobic sequencing batch reactor (ASBR). Ozonation increased BOD/COD by 37.9% with a COD loss of only 3.3%. At organic loads of 6.48-12.96 kg COD/m³/d, feeding with non-ozonated POME caused a system failure. The ozonated POME gave significantly higher TCOD removal at loadings 6.52 and 9.04 kg COD/m³/d but failed to sustain the operation at loading 11.67 kg COD/m³/d. Effects of cycle time (CT) and hydraulic retention time (HRT) were determined using quadratic regression model. The generated response surface and contour plot showed that at this high load conditions (6.52-11.67 kg COD/m³/d), longer HRT and shorter CT gave the ASBR higher organic removal efficiency and methane yield. The model was able to satisfactorily describe the relationship of these two key operating parameters.     

Improved fermentation performance to produce bioethanol from Gelidium amansii using Pichia stipitis adapted to galactose

Bioprocess and Biosystems Engineering - This study employed a statistical method to obtain optimal hyper thermal acid hydrolysis conditions using Gelidium amansii (red seaweed) as a source of...     

Effects of pH adjustment by parawood ash and effluent recycle ratio on the performance of anaerobic baffled reactors treating high sulfate wastewater

The objective of this study was to compare the performance of anaerobic baffled reactor (ABR) treating concentrated rubber latex wastewater under different pH adjustment substances and recycling ratios (R). Two ABRs, one received wastewater pretreated with NaOH and the other with ash, were operated at 35 degrees C under identical HRTs from 10 to 1.25d. Results show that both ABRs had highest COD and sulfate removal efficiencies at HRT 10d (averaged 82.71% and 96.16% of ABR-NaOH, and 80.77% and 96.60% of ABR-Ash, respectively), where majority of the influent COD and sulfate were removed by the first compartment of the ABR at all conditions tested. Increasing R (0, 0.3 and 0.5) raised the hydraulic loading on the system and resulted in a drop of organic removal efficiency and methane yield. Translocation of sulfate reducing bacteria and methanogens in the ABRs caused by increased organic loading and effluent recycle is discussed. The results show great potential of parawood ash as a pH adjustment substance for acidic wastewaters.     

Use of wood vinegar to enhance 5-aminolevulinic acid production by selected Rhodopseudomonas palustris in rubber sheet wastewater for agricultural use

This study aimed to produce inexpensive 5-aminolevulinic acid (ALA) in a non-sterile latex rubber sheet wastewater (RSW) by Rhodopseudomonas palustris TN114 and PP803 for the possibility to use in agricultural purposes by investigating the optimum conditions, and applying of wood vinegar (WV) as an economical source of levulinic acid to enhance ALA content. The Box–Behnken Design experiment was conducted under microaerobic-light conditions for 96 h with TN114, PP803 and their mixed culture (1:1) by varying initial pH, inoculum size (% v/v) and initial chemical oxygen demand (COD, mg/L). Results showed that the optimal condition (pH, % inoculum size, COD) of each set to produce extracellular ALA was found at 7.50, 6.00, 2000 for TN114; 7.50, 7.00, 3000 for PP803; and 7.50, 6.00, 4000 for a mixed culture; and each set achieved COD reduction as high as 63%, 71% and 75%, respectively. Addition of the optimal concentration of WV at mid log phase at 0.63% for TN114, and 1.25% for PP803 and the mixed culture significantly increased the ALA content by 3.7–4.2 times (128, 90 and 131 μM, respectively) compared to their controls. ALA production cost could be reduced approximately 31 times with WV on the basis of the amount of levulinic acid used. Effluent containing ALA for using in agriculture could be achieved by treating the RSW with the selected ALA producer R. palustris strains under the optimized condition with a little WV additive.     

Intensifying Clean Energy Production Through Cultivating Mixotrophic Microalgae from Digestates of Biogas Systems: Effects of Light Intensity,Medium Dilution,and Cultivating Time

High-strength wastewaters after being digested for biogas production in anaerobic digesters still contain substantial nutrients and organics. The anaerobic digestates from four major industries in Thailand were tested with batch cultivation of Chlorella sp. for oil production potentials. Pig farm digestate was found most suitable as the growth medium generating 0.95 g/L_medium (dry biomass), which was 1.16–3.06 times of other digestates tested. Considerable removals of nitrogen and phosphorus achieved were an added benefit to the goal of ultimate treatment of these wastewaters. Light intensity had strong influence on growth and heterotrophic metabolism up to 78 μmol/m²/s, while the dilution of digestate above 2.4× diminished growth potential and lipid production. A quadratic regression model was constructed to describe interaction of light intensity, dilution factor, and time of cultivation to lipid production with a satisfactory precision. Light intensity could influence fatty acid composition, although palmitic acid was found predominant at 47.1 %. The algae oil generated could potentially increase the total energy output from anaerobic digesters of a typical pig farm by 22 %.