• Mass Spectrometry Letters
• /
• v.4 no.2
• /
• pp.25-29
• /
• 2013

Proteomics for biomarker validation needs high throughput instrumentation to analyze huge set of clinical samples for quantitative and reproducible analysis at a minimum time without manual experimental errors. Sample preparation, a vital step in proteomics plays a major role in identification and quantification of proteins from biological samples. Tryptic digestion a major check point in sample preparation for mass spectrometry based proteomics needs to be more accurate with rapid processing time. The present study focuses on establishing a high throughput automated online system for proteolytic digestion and desalting of proteins from biological samples quantitatively and qualitatively in a reproducible manner. The present study compares online protein digestion and desalting of BSA with conventional off-line (in-solution) method and validated for real time sample for reproducibility. Proteins were identified using SEQUEST data base search engine and the data were quantified using IDEALQ software. The present study shows that the online system capable of handling high throughput samples in 96 well formats carries out protein digestion and peptide desalting efficiently in a reproducible and quantitative manner. Label free quantification showed clear increase of peptide quantities with increase in concentration with much linearity compared to off line method. Hence we would like to suggest that inclusion of this online system in proteomic pipeline will be effective in quantification of proteins in comparative proteomics were the quantification is really very crucial.

• Environmental Engineering Research
• /
• v.24 no.4
• /
• pp.662-669
• /
• 2019

Anaerobic digestion is a popular sewage sludge (Ss) treatment method as it provides significant pollution control and energy recovery. However, the low C/N ratio and poor biodegradability of Ss necessitate pretreatment methods that improve solubilization under anaerobic conditions in addition to anaerobic co-digestion with other substrates to improve the process efficiency. In this study, three pretreatment methods, namely microwave irradiation, ultrasonication, and heat treatment, were investigated, and the corresponding improvement in methane production was assessed. Additionally, the simplex centroid design method was utilized to determine the optimum mixture ratio of food waste (Fw), livestock manure (Lm), and Ss for maximum methane yield. Microwave irradiation at 700 W for 6 min yielded the highest biodegradability (62.0%), solubilization efficiency (59.7%), and methane production (329 mL/g VS). The optimum mixture ratio following pretreatment was 61.3% pretreated Ss, 28.6% Fw, and 10.1% Lm. The optimum mixture ratio without pretreatment was 33.6% un-pretreated Ss, 46.0% Fw, and 20.4% Lm. These results indicate that the choice of pretreatment method plays an important role in efficient anaerobic digestion and can be applied in operational plants to enhance methane production. Co-digestion of Ss with Fw and Lm was also beneficial.

• Journal of Korean Society for Atmospheric Environment
• /
• v.21 no.4
• /
• pp.423-430
• /
• 2005

This study was investigated with sample digestion techniques and a reliability of the analytical results for a quantitative analysis of a standard reference material ('urban particulate matter', SRM 1648). The metallic elements were analyzed by inductively coupled plasma mass spectrometry (ICP-MS) with closed vessel microwave acid digestion method and atomic absorption spectroscopy (AAS). Quality control of the overall analytical procedures for metallic element determinations of standard reference material were estimated by analysis of a SRM 1648. Three digestion solutions $(type\;1:\;HNO_3,\;type\;2:\;HNO_3\;and\;H2O_2(4'1)\;mixture,\;type\;3:5.55\%\; HNO_3\;and\;16.75\%\;HCI\;mixture)$ were applied to SRM 1648. As a result, three digestion solutions used in this study are completely unable to digest Cr of SRM 1648. Reliability of Cr and As showed some errors in the digestion with digestion solution type (3) due to the influence of chlorides. Type (3) digestion solutions are sufficient to fulfill the digestion of As as well as Se in SRM 1648. ICP-MS results showed the improvement in accurate and precise determination of some trace elements like Cd, V and Pb in SRM 1648. It is important to use the proper digestion solution for each element to qualify analytical precision.

• Environmental Analysis Health and Toxicology
• /
• v.27
• /
• pp.5.1-5.10
• /
• 2012

Objectives: The purpose of this study was to assess the risk of ingestion exposure of lead by particle sizes of crumb rubber in artificial turf filling material with consideration of bioavailability. Methods: This study estimated the ingestion exposure by particle sizes (more than 250 um or less than 250 um) focusing on recyclable ethylene propylene diene monomer crumb rubber being used as artificial turf filling. Analysis on crumb rubber was conducted using body ingestion exposure estimate method in which total content test method, acid extraction method and digestion extraction method are reflected. Bioavailability which is a calibrating factor was reflected in ingestion exposure estimate method and applied in exposure assessment and risk assessment. Two methods using acid extraction and digestion extraction concentration were compared and evaluated. Results: As a result of the ingestion exposure of crumb rubber material, the average lead exposure amount to the digestion extraction result among crumb rubber was calculated to be $1.56{\times}10^{-4}$ mg/kg-day for low grade elementary school students and $4.87{\times}10^{-5}$ mg/kg-day for middle and high school students in 250 um or less particle size, and that to the acid extraction result was higher than the digestion extraction result. Results of digestion extraction and acid extraction showed that the hazard quotient was estimated by about over 2 times more in particle size of lower than 250 um than in higher than 250 um. There was a case of an elementary school student in which the hazard quotient exceeded 0.1. Conclusions: Results of this study confirm that the exposure of lead ingestion and risk level increases as the particle size of crumb rubber gets smaller.

• Animal Bioscience
• /
• v.35 no.10
• /
• pp.1592-1605
• /
• 2022

Objective: The objective of this study was to select an effective in vitro digestion-fermentation model to estimate the effect of decreasing dietary crude protein (CP) on odor emission during pig production and to suggest potential prediction markers through in vitro and in vivo experiments. Methods: In the in vitro experiment, three diet formulations with different CP contents (170 g/kg, 150 g/kg, and 130 g/kg) but containing the same standardized ileal digestible essential amino acids (SID-EAA) were assessed. Each diet was evaluated by two different in vitro gastric-intestinal phase digestion methods (flask and dialysis), combined with fresh pig feces-ferment inoculation. Eighteen growing barrows (31.9±1.6 kg) were divided into three groups: control diet (180 g CP/kg, without SID-EAA adjustment), 170 g CP/kg diet, and 150 g CP/kg diet for 4 weeks. Results: The in vitro digestion results indicated that in vitro digestibility was affected by the gastric-intestinal phase digestion method and dietary CP level. According to the gas kinetic and digestibility results, the dialysis method showed greater distinguishability for dietary CP level adjustment. Nitrogen-related odor compounds (NH₃-N, indole, p-cresol, and skatole) were highly correlated with urease and protease activity. The feeding study indicated that both EAA-adjusted diets resulted in a lower odor emission especially in p-cresol and skatole. Both protease and urease activity in feces were also closely related to odor emissions from nitrogen metabolism compounds. Conclusion: Dialysis digestion in the gastric-intestinal phase followed by fresh fecal inoculation fermentation is suitable for in vitro diet evaluation. The enzyme activity in the fermentation and the fecal samples might provide a simple and effective estimation tool for nitrogen-related odor emission prediction in both in vitro and in vivo experiments.

• Korean Journal of Environmental Agriculture
• /
• v.30 no.1
• /
• pp.31-36
• /
• 2011

BACKGROUND: Disposal of slurry animal manure produced by an anaerobic slurry-type barn method is not easy since the animal slurry contain high moisture content which makes solid-liquid separation a difficult process. However, recently, the interest about anaerobic digestion process as an environment-friendly waste disposal method has gained a wide interest because it can treat highly organic matter contained by the piggery slurry, decrease the odor after treatment, and enable the effective recovery of the methane gas which is a valuable energy resource. The objectives of this study were to identify the solubilization characteristics and to improve the anaerobic digestion efficiency of piggery slurry through full-scale anaerobic digestion experiments. METHODS AND RESULTS: In a full-scale continuous anaerobic digestion operation, the adaptability of single anaerobic digestion and its digestion efficiency were also evaluated in the farm field. The actual pH range and alkalinity concentration of piggery slurry used during the operation were comparatively higher than the concentrations of pH and alkalinity in the digestion tank which were stable at 7.5~8.0, 4,008 mg/L (as$CaCO_3$), respectively. The removal efficiency of organic matter (TCOD) by anaerobic digestion was 75~90% and methane gas production amount was at 0.33 L/L/day, a little higher than that of ordinary animal manure. CONCLUSION(s): Our findings showed higher recovery of highly purified methane and greater efficiency of anaerobic tank digestion since its methane gas content was at 65~70%.

• Korean Journal of Veterinary Research
• /
• v.44 no.2
• /
• pp.269-277
• /
• 2004

Trichinella spiralis is one of the important zoonotic parasites with a wide variety of vertebrates hosts in nature. The purpose of this study were to analyze ESP(Excretory-Secretory Protein) antigen, to evaluate ELISA for the serological diagnosis of Trichinosis, and to survey T. spiralis infection in finishing pigs using the pepsin digestion method and ELISA in Korea. In the analysis of ESP antigen by SDS-PAGE and Western blot, 4 major bands (70, 55, 52.6, and 49 kDa) were revealed from the ESP antigen. Predilection sites of T. spiralis were the diaphragm, the tongue, masseter muscles, intercostal muscle, and hindlimb in orders in the experimentally infected rats. Sera from 581 swine were tested by ELISA with ESP antigen. The 54 (9.3%) sera were suspected as positive reactors, however, these 54 sera were determined as false positives by the use of Western blotting. This study demonstrated that the ELISA was not suitable for the examination of T. spiralis in pork. The diaphragm muscle samples of 251 finishing pigs were tested by the method of pepsin-digestion for the presence of Trichinella larvae, however, T. spiralis was not detected from the samples. We could not find out T. spiralis infection in pig in Korea pork.

• Advances in environmental research
• /
• v.5 no.4
• /
• pp.251-262
• /
• 2016

The biodegradability and decomposition efficiencies increase with the pre-treatment of sludge in a digestion process. In this study, the feasibility of ultrasound coupled with potassium permanganate oxidation as a disintegration method and digestibility of aerobic reactor fed with disintegrated sludge with ultrasound coupled potassium permanganate were investigated. The first stage of the study focused on determining the optimum condition for ultrasonic pre-treatment for achieving better destruction efficiency of sludge. The second part of the study, the aerobic digestibility of sludge disintegrated with ultrasound and potassium permanganate oxidation alone and combined were examined comparatively. The results showed that when 20 min of ultrasonic pre-treatment applied, the specific energy output was 49384 kJ/kgTS with disintegration degree of 58.84%. During the operation of aerobic digester, VS/TS ratios of digesters fed with disintegrated sludge decreased indicating that disintegration methods could obviously enhance aerobic digestion performance. The highest reduction in volatile solids was 75% in the digester fed with ultrasound+potassium permanganate disintegrated sludge at the end of the operation compared to digester fed with raw sludge. Total Nitrogen (TN) and Total Phosphorus (TP) levels in sludge supernatant increased with this combined method significantly. Besides, it promoted the production of ${\bullet}OH$, thus enhancing the release of Carbon (C), Nitrogen (N) and Phosphorus (P) from the sludge. Disintegration with all methods used in this study could not improve Capillary Suction Time (CST) reduction in disintegrated digesters during the operation. The results demonstrated that the combined ultrasound treatment and potassium permanganate oxidation method improves the biodegradability compared to control reactor or their single application.

• Asian-Australasian Journal of Animal Sciences
• /
• v.15 no.11
• /
• pp.1659-1676
• /
• 2002

Ruminant animals develop a diverse and sophisticated microbial ecosystem for digesting fibrous feedstuffs. Plant cell walls are complex and their structures are not fully understood, but it is generally believed that the chemical properties of some plant cell wall compounds and the cross-linked three-dimensional matrix of polysaccharides, lignin and phenolic compounds limit digestion of cell wall polysaccharides by ruminal microbes. Three adaptive strategies have been identified in the ruminal ecosystem for degrading plant cell walls: production of the full slate of enzymes required to cleave the numerous bonds within cell walls; attachment and colonization of feed particles; and synergetic interactions among ruminal species. Nonetheless, digestion of fibrous feeds remains incomplete, and numerous research attempts have been made to increase this extent of digestion. Exogenous fibrolytic enzymes (EFE) have been used successfully in monogastric animal production for some time. The possibility of adapting EFE as feed additives for ruminants is under intensive study. To date, animal responses to EFE supplements have varied greatly due to differences in enzyme source, application method, and types of diets and livestock. Currently available information suggests delivery of EFE by applying them to feed offers the best chance to increase ruminal digestion. The general tendency of EFE to increase rate, but not extent, of fibre digestion indicates that the products currently on the market for ruminants may not be introducing novel enzyme activities into the rumen. Recent research suggests that cleavage of esterified linkages (e.g., acetylesterase, ferulic acid esterase) within the plant cell wall matrix may be the key to increasing the extent of cell wall digestion in the rumen. Thus, a crucial ingredient in an effective enzyme additive for ruminants may be an as yet undetermined esterase that may not be included, quantified or listed in the majority of available enzyme preparations. Identifying these pivotal enzyme(s) and using biotechnology to enhance their production is necessary for long term improvements in feed digestion using EFE. Pretreating fibrous feeds with alkali in addition to EFE also shows promise for improving the efficacy of enzyme supplements.

• Journal of Animal Science and Technology
• /
• v.66 no.2
• /
• pp.374-386
• /
• 2024

To predict the apparent total tract digestibility (ATTD) of crude protein (CP) in dogs we developed an in vitro system using an in vitro digestion method and a statistical analysis. The experimental diets used chicken meat powder as the protein source, with CP levels of 20% (22.01%, analyzed CP value as dry-based), 30% (31.35%, analyzed CP value as dry-based), and 40% (41.34%, analyzed CP value as dry-based). To simulate in vivo digestive processes a static in vitro digestion was performed in two steps; stomach and small intestine. To analyze ATTD the total fecal samples were collected in eight neutered beagle dogs during the experimental period. CP digestibility was calculated by measuring CP levels in dog food, in vitro undigested fraction, and dog feces. In result, CP digestibility at both in vivo and in vitro was increased with increasing dietary CP levels. To estimate in vivo digestibility the co-relation of in vivo ATTD and in vitro digestibility was investigated statistically and a regression equation was developed to predict the CP ATTD (% = 2.5405 × in vitro CP digestibility (%) + + 151.8). The regression equation was evaluated its feasibility by using a commercial diet. The predicted CP digestibility which was calculated by the regression equation showed high index of similarity (100.16%) with that of in vivo in dogs. With that, it would be a feasible non-animal method to predict in vivo CP digestibility by using in vitro digestion method and the proposed linear regression equation in adult dogs.