Kidney Dysfunction in Adult Offspring Exposed In Utero to Type 1 Diabetes Is Associated with Alterations in Genome-Wide DNA Methylation

PubMed Central

Gautier, Jean-François; Porcher, Raphaël; Abi Khalil, Charbel; Bellili-Munoz, Naima; Fetita, Lila Sabrina; Travert, Florence; Choukem, Simeon-Pierre; Riveline, Jean-Pierre; Hadjadj, Samy; Larger, Etienne; Boudou, Philippe; Blondeau, Bertrand; Roussel, Ronan; Ferré, Pascal; Ravussin, Eric; Rouzet, François; Marre, Michel

2015-01-01

Background Fetal exposure to hyperglycemia impacts negatively kidney development and function. Objective Our objective was to determine whether fetal exposure to moderate hyperglycemia is associated with epigenetic alterations in DNA methylation in peripheral blood cells and whether those alterations are related to impaired kidney function in adult offspring. Design Twenty nine adult, non-diabetic offspring of mothers with type 1 diabetes (T1D) (case group) were matched with 28 offspring of T1D fathers (control group) for the study of their leukocyte genome-wide DNA methylation profile (27,578 CpG sites, Human Methylation 27 BeadChip, Illumina Infinium). In a subset of 19 cases and 18 controls, we assessed renal vascular development by measuring Glomerular Filtration Rate (GFR) and Effective Renal Plasma Flow (ERPF) at baseline and during vasodilatation produced by amino acid infusion. Results Globally, DNA was under-methylated in cases vs. controls. Among the 87 CpG sites differently methylated, 74 sites were less methylated and 13 sites more methylated in cases vs. controls. None of these CpG sites were located on a gene known to be directly involved in kidney development and/or function. However, the gene encoding DNA methyltransferase 1 (DNMT1)—a key enzyme involved in gene expression during early development–was under-methylated in cases. The average methylation of the 74 under-methylated sites differently correlated with GFR in cases and controls. Conclusion Alterations in methylation profile imprinted by the hyperglycemic milieu of T1D mothers during fetal development may impact kidney function in adult offspring. The involved pathways seem to be a nonspecific imprinting process rather than specific to kidney development or function. PMID:26258530

Kidney Dysfunction in Adult Offspring Exposed In Utero to Type 1 Diabetes Is Associated with Alterations in Genome-Wide DNA Methylation.

PubMed

Gautier, Jean-François; Porcher, Raphaël; Abi Khalil, Charbel; Bellili-Munoz, Naima; Fetita, Lila Sabrina; Travert, Florence; Choukem, Simeon-Pierre; Riveline, Jean-Pierre; Hadjadj, Samy; Larger, Etienne; Boudou, Philippe; Blondeau, Bertrand; Roussel, Ronan; Ferré, Pascal; Ravussin, Eric; Rouzet, François; Marre, Michel

2015-01-01

Fetal exposure to hyperglycemia impacts negatively kidney development and function. Our objective was to determine whether fetal exposure to moderate hyperglycemia is associated with epigenetic alterations in DNA methylation in peripheral blood cells and whether those alterations are related to impaired kidney function in adult offspring. Twenty nine adult, non-diabetic offspring of mothers with type 1 diabetes (T1D) (case group) were matched with 28 offspring of T1D fathers (control group) for the study of their leukocyte genome-wide DNA methylation profile (27,578 CpG sites, Human Methylation 27 BeadChip, Illumina Infinium). In a subset of 19 cases and 18 controls, we assessed renal vascular development by measuring Glomerular Filtration Rate (GFR) and Effective Renal Plasma Flow (ERPF) at baseline and during vasodilatation produced by amino acid infusion. Globally, DNA was under-methylated in cases vs. controls. Among the 87 CpG sites differently methylated, 74 sites were less methylated and 13 sites more methylated in cases vs. controls. None of these CpG sites were located on a gene known to be directly involved in kidney development and/or function. However, the gene encoding DNA methyltransferase 1 (DNMT1)--a key enzyme involved in gene expression during early development--was under-methylated in cases. The average methylation of the 74 under-methylated sites differently correlated with GFR in cases and controls. Alterations in methylation profile imprinted by the hyperglycemic milieu of T1D mothers during fetal development may impact kidney function in adult offspring. The involved pathways seem to be a nonspecific imprinting process rather than specific to kidney development or function.

Changes in solvation during DNA binding and cleavage are critical to altered specificity of the EcoRI endonuclease

PubMed Central

Robinson, Clifford R.; Sligar, Stephen G.

1998-01-01

Restriction endonucleases such as EcoRI bind and cleave DNA with great specificity and represent a paradigm for protein–DNA interactions and molecular recognition. Using osmotic pressure to induce water release, we demonstrate the participation of bound waters in the sequence discrimination of substrate DNA by EcoRI. Changes in solvation can play a critical role in directing sequence-specific DNA binding by EcoRI and are also crucial in assisting site discrimination during catalysis. By measuring the volume change for complex formation, we show that at the cognate sequence (GAATTC) EcoRI binding releases about 70 fewer water molecules than binding at an alternate DNA sequence (TAATTC), which differs by a single base pair. EcoRI complexation with nonspecific DNA releases substantially less water than either of these specific complexes. In cognate substrates (GAATTC) kcat decreases as osmotic pressure is increased, indicating the binding of about 30 water molecules accompanies the cleavage reaction. For the alternate substrate (TAATTC), release of about 40 water molecules accompanies the reaction, indicated by a dramatic acceleration of the rate when osmotic pressure is raised. These large differences in solvation effects demonstrate that water molecules can be key players in the molecular recognition process during both association and catalytic phases of the EcoRI reaction, acting to change the specificity of the enzyme. For both the protein–DNA complex and the transition state, there may be substantial conformational differences between cognate and alternate sites, accompanied by significant alterations in hydration and solvent accessibility. PMID:9482860

Comparison of the nucleotide and amino acid sequences of the RsrI and EcoRI restriction endonucleases.

PubMed

Stephenson, F H; Ballard, B T; Boyer, H W; Rosenberg, J M; Greene, P J

1989-12-21

The RsrI endonuclease, a type-II restriction endonuclease (ENase) found in Rhodobacter sphaeroides, is an isoschizomer of the EcoRI ENase. A clone containing an 11-kb BamHI fragment was isolated from an R. sphaeroides genomic DNA library by hybridization with synthetic oligodeoxyribonucleotide probes based on the N-terminal amino acid (aa) sequence of RsrI. Extracts of E. coli containing a subclone of the 11-kb fragment display RsrI activity. Nucleotide sequence analysis reveals an 831-bp open reading frame encoding a polypeptide of 277 aa. A 50% identity exists within a 266-aa overlap between the deduced aa sequences of RsrI and EcoRI. Regions of 75-100% aa sequence identity correspond to key structural and functional regions of EcoRI. The type-II ENases have many common properties, and a common origin might have been expected. Nevertheless, this is the first demonstration of aa sequence similarity between ENases produced by different organisms.

Construction of a novel gene bank of Bacillus subtilis using a low copy number vector in Escherichia coli.

PubMed

Hasnain, S; Thomas, C M

1986-07-01

Low copy number vector plasmid pCT571 was constructed to clone Bacillus subtilis genomic fragments in Escherichia coli. pCT571 confers KmR, TcR and CmR in E. coli and CmR in B. subtilis. It has unique restriction sites within the KmR and TcR markers to allow screening for recombinant plasmids by insertional inactivation of these genes. It contains the pSC101 replicon and replicates normally at six to eight copies per chromosome equivalent in E. coli. It also contains oriVRK2, which when supplied with the product of the trfA gene of RK2 in trans, allows pCT571 to replicate at 35-40 copies per chromosome equivalent. A B. subtilis gene bank was created by cloning partially Sau3A-digested and size-fractionated fragments of B. subtilis chromosomal DNA into the BamHI site of pCT571. DNA from 1097 KmR TcS transformants was extracted and analysed electrophoretically as supercoiled DNA and after digesting with EcoRI or EcoRI and SalI. Approximately 1000 hybrid plasmids were found with reasonably sized B. subtilis fragments. The mean size of the inserts in pCT571 is 8 kb, ranging from 4 to 20 kb in different plasmids. The gene bank covers most of the B. subtilis chromosome, as demonstrated by the results of screening the gene bank for selectable nutritional markers in E. coli and B. subtilis. Hybrid plasmids which complement E. coli mutants for arg, his, lys, met, pdx, pyr and thr markers were identified from the gene bank. In B. subtilis the presence of argC, cysA, dal, hisA, ilvA, leuA, lys, metB, metC, phe, purA, purB, thr and trpC was established by transformation experiments. The effects of copy number on cloning and long-term maintenance in the bacterial strains were also investigated. At high copy number some hybrid plasmids cannot be maintained at all, while others show an increased rate of structural deletions and rearrangements.

No clustering for linkage map based on low-copy and undermethylated microsatellites.

PubMed

Zhou, Yi; Gwaze, David P; Reyes-Valdés, M Humberto; Bui, Thomas; Williams, Claire G

2003-10-01

Clustering has been reported for conifer genetic maps based on hypomethylated or low-copy molecular markers, resulting in uneven marker distribution. To test this, a framework genetic map was constructed from three types of microsatellites: low-copy, undermethylated, and genomic. These Pinus taeda L. microsatellites were mapped using a three-generation pedigree with 118 progeny. The microsatellites were highly informative; of the 32 markers in intercross configuration, 29 were segregating for three or four alleles in the progeny. The sex-averaged map placed 51 of the 95 markers in 15 linkage groups at LOD > 4.0. No clustering or uneven distribution across the genome was observed. The three types of P. taeda microsatellites were randomly dispersed within each linkage group. The 51 microsatellites covered a map distance of 795 cM, an average distance of 21.8 cM between markers, roughly half of the estimated total map length. The minimum and maximum distances between any two bins was 4.4 and 45.3 cM, respectively. These microsatellites provided anchor points for framework mapping for polymorphism in P. taeda and other closely related hard pines.

DNA hypomethylation of individual sequences in aborted cloned bovine fetuses.

PubMed

Chen, Tao; Jiang, Yan; Zhang, Yan-Ling; Liu, Jing-He; Hou, Yi; Schatten, Heide; Chen, Da-Yuan; Sun, Qing-Yuan

2005-09-01

Cloned bovines have a much higher abortion rate than those derived in vivo. Available evidence indicates that inappropriate epigenetic reprogramming of donor nuclei is the primary cause of cloning failure. To gain a better understanding of the DNA methylation changes associated with the high abortion rate of cloned bovines, we examined the DNA methylation status of a repeated sequence (satellite I) and the promoter regions of two single-copy genes (interleukin 3/cytokeratin) in aborted cloned fetuses, aborted fetuses derived from artificial insemination (AI), cloned adults and AI adults by bisulfite sequencing and restriction enzyme analysis. Two of four aborted cloned fetuses show very low methylation levels in the two single-copy gene promoter regions. One of the two fetuses also showed undermethylated status in the satellite I sequence. The other two aborted cloned fetuses have similar methylation levels to those of aborted AI fetuses. However, no difference in methylation was observed between cloned adults and AI adults. Our results demonstrate for the first time the undermethylated status of individual sequences in aborted cloned fetuses. These findings suggest that aberrant DNA methylation may contribute to the developmental failure of cloned bovine fetuses.

Nickel-Catalyzed Proton-Deuterium Exchange (HDX) Procedures for Glycosidic Linkage Analysis of Complex Carbohydrates.

PubMed

Price, Neil P J; Hartman, Trina M; Vermillion, Karl E

2015-07-21

The structural analysis of complex carbohydrates typically requires the assignment of three parameters: monosaccharide composition, the position of glycosidic linkages between monosaccharides, and the position and nature of noncarbohydrate substituents. The glycosidic linkage positions are often determined by permethylation analysis, but this can be complicated by high viscosity or poor solubility, resulting in under-methylation. This is a drawback because an under-methylated position may be misinterpreted as the erroneous site of a linkage or substituent. Here, we describe an alternative approach to linkage analysis that makes use of a nonreversible deuterium exchange of C-H protons on the carbohydrate backbone. The exchange reaction is conducted in deuterated water catalyzed by Raney nickel, and results in the selective exchange of C-H protons adjacent to free hydroxyl groups. Hence, the position of the residual C-H protons is indicative of the position of glycosidic linkages or other substituents and can be readily assigned by heteronuclear single quantum coherence-nuclear magnetic resonance (HSQC-NMR) or, following suitable derivatization, by gas chromatography-mass spectroscopy (GC/MS) analysis. Moreover, because the only changes to the parent sugar are proton/deuterium exchanges, the composition and linkage analysis can be determined in a single step.

Mitochondrial DNA variation and genetic relationships of Populus species.

PubMed

Barrett, J W; Rajora, O P; Yeh, F C; Dancik, B P; Strobeck, C

1993-02-01

We examined variation in and around the region coding for the cytochrome c oxidase I (coxI) and ATPase 6 (atp6) genes in the mitochondrial genomes of four Populus species (P. nigra, P. deltoides, P. maximowiczii, and P. tremuloides) and the natural hybrid P. x canadensis (P. deltoides x P. nigra). Total cellular DNAs of these poplars were digested with 16 restriction endonucleases and probed with maize mtDNA-specific probes (CoxI and Atp6). The only variant observed for Atp6 was interspecific, with P. maximowiczii separated from the other species as revealed by EcoRI digestions. No intraspecific mtDNA variation was observed among individuals of P. nigra, P. maximowiczii, P. x canadensis, or P. tremuloides for the CoxI probe. However, two varieties of P. deltoides were distinct because of a single site change in the KpnI digestions, demonstrating that P. deltoides var. deltoides (eastern cottonwood) and var. occidentalis (plains cottonwood) have distinct mitochondrial genomes in the region of the coxI gene. Populus x canadensis shared the same restriction fragment patterns as its suspected maternal parent P. deltoides. Nucleotide substitutions per base in and around the coxI and atp6 genes among the Populus species and the hybrid ranged from 0.0017 to 0.0077. The interspecific estimates of nucleotide substitution per base suggested that P. tremuloides was furthest removed from P. deltoides and P. x canadensis and least diverged from P. nigra. Populus maximowiczii was placed between these two clusters.

Characterization of two nisin-producing Lactococcus lactis subsp. lactis strains isolated from a commercial sauerkraut fermentation.

PubMed Central

Harris, L J; Fleming, H P; Klaenhammer, T R

1992-01-01

Two Lactococcus lactis subsp. lactis strains, NCK400 and LJH80, isolated from a commercial sauerkraut fermentation were shown to produce nisin. LJH80 was morphologically unstable and gave rise to two stable, nisin-producing (Nip+) derivatives, NCK318-2 and NCK318-3. NCK400 and derivatives of LJH80 exhibited identical morphological and metabolic characteristics, but could be distinguished on the basis of plasmid profiles and genomic hybridization patterns to a DNA probe specific for the iso-ISS1 element, IS946. NCK318-2 and NCK318-3 harbored two and three plasmids, respectively, which hybridized with IS946. Plasmid DNA was not detected in NCK400, and DNA from this strain failed to hybridize with IS946. Despite the absence of detectable plasmid DNA in NCK400, nisin-negative derivatives (NCK402 and NCK403) were isolated after repeated transfer in broth at 37 degrees C. Nisin-negative derivatives concurrently lost the ability to ferment sucrose and became sensitive to nisin. A 4-kbp HindIII fragment containing the structural gene for nisin (spaN), cloned from L. lactis subsp. lactis ATCC 11454, was used to probe genomic DNA of NCK318-2, NCK318-3, NCK400, and NCK402 digested with EcoRI or HindIII. The spaN probe hybridized to an 8.8-kbp EcoRI fragment and a 10-kbp HindIII fragment in the Nip+ sauerkraut isolates, but did not hybridize to the Nip- derivative, NCK402. A different hybridization pattern was observed when the same probe was used against Nip+ L. lactis subsp. lactis ATCC 11454 and ATCC 7962. These phenotypic and genetic data confirmed that unique Nip+ L. lactis subsp. lactis strains were isolated from fermenting sauerkraut. Images PMID:1622214

Molecular cloning and expression of Corynebacterium glutamicum genes for amino acid synthesis in Escherichia coli cells

DOE Office of Scientific and Technical Information (OSTI.GOV)

Beskrovnaya, O.Yu.; Fonshtein, M.Yu.; Kolibaba, L.G.

1989-01-01

Molecular cloning of Corynebacterium glutamicum genes for threonine and lysine synthesis has been done in Escherichia coli cells. The clonal library of EcoRI fragments of chromosomal DNA of C. glutamicum was constructed on the plasmid vector /lambda/pSL5. The genes for threonine and lysine synthesis were identified by complementation of E. coli mutations in thrB and lysA genes, respectively. Recombinant plasmids, isolated from independent ThrB/sup +/ clone have a common 4.1-kb long EcoRI DNA fragment. Hybrid plasmids isolated from LysA/sup +/ transductants of E. coli have common 2.2 and 3.3 kb long EcoRI fragments of C. glutamicum DNA. The hybrid plasmidsmore » consistently transduced the markers thrB/sup +/ and lysA/sup +/. The Southern hybridization analysis showed that the cloned DNA fragments hybridized with the fragments of identical length in C. glutamicum chromosomes.« less

Distributed biotin–streptavidin transcription roadblocks for mapping cotranscriptional RNA folding

PubMed Central

Strobel, Eric J.; Nedialkov, Yuri; Artsimovitch, Irina

2017-01-01

Abstract RNA folding during transcription directs an order of folding that can determine RNA structure and function. However, the experimental study of cotranscriptional RNA folding has been limited by the lack of easily approachable methods that can interrogate nascent RNA structure at nucleotide resolution. To address this, we previously developed cotranscriptional selective 2΄-hydroxyl acylation analyzed by primer extension sequencing (SHAPE-Seq) to simultaneously probe all intermediate RNA transcripts during transcription by stalling elongation complexes at catalytically dead EcoRIE111Q roadblocks. While effective, the distribution of elongation complexes using EcoRIE111Q requires laborious PCR using many different oligonucleotides for each sequence analyzed. Here, we improve the broad applicability of cotranscriptional SHAPE-Seq by developing a sequence-independent biotin–streptavidin (SAv) roadblocking strategy that simplifies the preparation of roadblocking DNA templates. We first determine the properties of biotin–SAv roadblocks. We then show that randomly distributed biotin–SAv roadblocks can be used in cotranscriptional SHAPE-Seq experiments to identify the same RNA structural transitions related to a riboswitch decision-making process that we previously identified using EcoRIE111Q. Lastly, we find that EcoRIE111Q maps nascent RNA structure to specific transcript lengths more precisely than biotin–SAv and propose guidelines to leverage the complementary strengths of each transcription roadblock in cotranscriptional SHAPE-Seq. PMID:28398514

Distributed biotin-streptavidin transcription roadblocks for mapping cotranscriptional RNA folding.

PubMed

Strobel, Eric J; Watters, Kyle E; Nedialkov, Yuri; Artsimovitch, Irina; Lucks, Julius B

2017-07-07

RNA folding during transcription directs an order of folding that can determine RNA structure and function. However, the experimental study of cotranscriptional RNA folding has been limited by the lack of easily approachable methods that can interrogate nascent RNA structure at nucleotide resolution. To address this, we previously developed cotranscriptional selective 2΄-hydroxyl acylation analyzed by primer extension sequencing (SHAPE-Seq) to simultaneously probe all intermediate RNA transcripts during transcription by stalling elongation complexes at catalytically dead EcoRIE111Q roadblocks. While effective, the distribution of elongation complexes using EcoRIE111Q requires laborious PCR using many different oligonucleotides for each sequence analyzed. Here, we improve the broad applicability of cotranscriptional SHAPE-Seq by developing a sequence-independent biotin-streptavidin (SAv) roadblocking strategy that simplifies the preparation of roadblocking DNA templates. We first determine the properties of biotin-SAv roadblocks. We then show that randomly distributed biotin-SAv roadblocks can be used in cotranscriptional SHAPE-Seq experiments to identify the same RNA structural transitions related to a riboswitch decision-making process that we previously identified using EcoRIE111Q. Lastly, we find that EcoRIE111Q maps nascent RNA structure to specific transcript lengths more precisely than biotin-SAv and propose guidelines to leverage the complementary strengths of each transcription roadblock in cotranscriptional SHAPE-Seq. © The Author(s) 2017. Published by Oxford University Press on behalf of Nucleic Acids Research.

Characterization of the repetitive DNA elements in the genome of fish lymphocystis disease viruses.

PubMed

Schnitzler, P; Darai, G

1989-09-01

The complete DNA nucleotide sequence of the repetitive DNA elements in the genome of fish lymphocystis disease virus (FLDV) isolated from two different species (flounder and dab) was determined. The size of these repetitive DNA elements was found to be 1413 bp which corresponds to the DNA sequences of the 5' terminus of the EcoRI DNA fragment B (0.034 to 0.052 m.u.) and to the EcoRI DNA fragment M (0.718 to 0.736 m.u.) of the FLDV genome causing lymphocystis disease in flounder and plaice. The degree of DNA nucleotide homology between both regions was found to be 99%. The repetitive DNA element in the genome of FLDV isolated from other fish species (dab) was identified and is located within the EcoRI DNA fragment B and J of the viral genome. The DNA nucleotide sequence of one duplicate of this repetition (EcoRI DNA fragment J) was determined (1410 bp) and compared to the DNA nucleotide sequences of the repetitive DNA elements of the genome of FLDV isolated from flounder. It was found that the repetitive DNA elements of the genome of FLDV derived from two different fish species are highly conserved and possess a degree of DNA sequence homology of 94%. The DNA sequences of each strand of the individual repetitive element possess one open reading frame.

A tag-based approach for high-throughput analysis of CCWGG methylation.

PubMed

Denisova, Oksana V; Chernov, Andrei V; Koledachkina, Tatyana Y; Matvienko, Nicholas I

2007-10-15

Non-CpG methylation occurring in the context of CNG sequences is found in plants at a large number of genomic loci. However, there is still little information available about non-CpG methylation in mammals. Efficient methods that would allow detection of scarcely localized methylated sites in small quantities of DNA are required to elucidate the biological role of non-CpG methylation in both plants and animals. In this study, we tested a new whole genome approach to identify sites of CCWGG methylation (W is A or T), a particular case of CNG methylation, in genomic DNA. This technique is based on digestion of DNAs with methylation-sensitive restriction endonucleases EcoRII-C and AjnI. Short DNAs flanking methylated CCWGG sites (tags) are selectively purified and assembled in tandem arrays of up to nine tags. This allows high-throughput sequencing of tags, identification of flanking regions, and their exact positions in the genome. In this study, we tested specificity and efficiency of the approach.

Mechanism for circularization of linear DNAs: circular parvovirus MVM DNA is formed by a "noose" sliding in a "lasso"-like DNA structure.

PubMed

Bratosin, S; Laub, O; Tal, J; Aloni, Y

1979-09-01

During an electron-microscopic survey with the aim of identifying the parvovirus MVM transcription template, we observed previously unidentified structures of MVM DNA in lysates of virus-infected cells. These included double-stranded "lasso"-like structures and relaxed circles. Both structures were of unit length MVM DNA, indicating that they were not intermediates formed during replication; they each represented about 5% of the total nuclear MVM DNA. The proportion of these structures was unchanged after digestion with sodium dodecyl sulfate/Pronase and RNase and after mild denaturation treatment. Cleavage of the "lasso" structures with EcoRI restriction endonuclease indicated that the "noose" part of the "lasso" structure is located on the 5' side of the genomic single-stranded MVM DNA. A model is presented for the molecular nature of the circularization process of MVM DNA in which the "lasso" structures are identified as intermediates during circle formation. This model proposes a mechanism for circularization of linear DNAs.

Restriction enzyme analysis of Indian isolates of egg drop syndrome 1976 virus recovered from chicken, duck and quail.

PubMed

Senthilkumar, N; Kataria, J M; Koti, M; Dhama, K; Dash, B B

2004-07-01

Egg drop syndrome 1976 (EDS-76) is caused by a haemagglutinating adenovirus belonging to group III of the genus Aviadenovirus in the family Adenoviridae. All isolates are serologically identical, but have been divided into three groups based on restriction endonuclease (RE) analysis. In this study the viral DNA of various Indian EDS-76 viral isolates (CEDS-A, CEDS-B, EDS-M, EDS-ML, EDS-1/AD/86, EDS-KC and QEDS) obtained from different avian species and different geographical regions were digested with restriction endonucleases viz., EcoRI, BamHI, HindIII and PstI. The results showed that one Indian isolate obtained from duck (DEDS-KC) was different from all other chicken and quail counterparts. All other isolates were identical to the reference viral strain BC-14, which belong to group I of EDS-76 viruses. The duck isolate EDS-KC could not be placed in any of the three groups reported earlier.

Person-to-person transfer of Candida albicans in the spacecraft environment

NASA Technical Reports Server (NTRS)

Pierson, D. L.; Mehta, S. K.; Magee, B. B.; Mishra, S. K.

1995-01-01

We assessed the exchange of Candida albicans among crew members during 10 Space Shuttle missions. Throat, nasal, urine and faecal specimens were collected from 61 crew members twice before and once after space flights ranging from 7 to 10 days in duration; crews consisted of groups of five, six or seven men and women. Candida albicans was isolated at least once from 20 of the 61 subjects (33%). Candida strains were identified by restriction-fragment length polymorphism (RFLP) after digestion by the endonucleases EcoRI and HinfI; further discrimination was gained by Southern blot hybridization with the C. albicans repeat fragment 27A. Eighteen of the 20 Candida-positive crew members carried different strains of C. albicans in the specimens collected. Possible transfer of C. albicans between members of the same crew was demonstrated only once in the 10 missions studied. We conclude that the transfer of C. albicans among crew members during Space Shuttle flights is less frequent than had been predicted from earlier reports.

A Novel Locomotion-based Validation Assay for Candidate Drugs Using Drosophila DYT1 Disease Model

DTIC Science & Technology

2014-06-01

rescue the locomotion defects of Drosophila larvae caused by the expression of human torsinAΔE. These results demonstrated that human torsinA can... Drosophila dtorsin∆D transgenic lines dtorsin∆E and dtorsin∆D cDNA constructs were made from the wild type dtorsin cDNA using QuikChange II XL Site...After confirming mutated sequences , the insert was again cut out with EcoRI and NotI and inserted between EcoRI and NotI sites of pUAST [2] to produce

Ribotype diversity of Listeria monocytogenes isolates from two salmon processing plants in Norway.

PubMed

Klaeboe, Halvdan; Rosef, Olav; Fortes, Esther; Wiedmann, Martin

2006-10-01

The purpose of this study was to use automated ribotyping procedure to track Listeria monocytogenes transmission in the cold smoked fish production chain and to characterize L. monocytogenes subtypes associated with the salmon processing industry. A total of 104 isolates, which had previously been obtained from a raw fish slaughter and processing plant (plant B) and an adjacent, downstream, salmon smoking operation (plant A), were characterized. These isolates had been obtained through a longitudinal study on Listeria presence, which covered a 31-week period, in both plants. Isolates had been obtained from samples taken from different machinery used throughout the production process. In addition, six isolates obtained from products produced in plant A two years after the initial study were included, so that a total of 110 isolates were characterized. Automated ribotyping was performed using both the restriction enzymes EcoRI and PvuII to increase the discriminatory power. The 110 L. monocytogenes isolates could be divided into 11 EcoRI ribotypes; PvuII ribotype data yielded multiple subtypes within 7 EcoRI ribotypes for a total of 21 subtypes based on both EcoRI and PvuII ribotyping. A total of three EcoRI ribotypes (DUP-1023C, DUP-1045B, and DUP-1053E) were isolated at multiple sampling times from both plants. In addition, one subtype (DUP-1053B) was isolated at multiple sampling times in only plant A, the salmon smoking operation. These data not only support that L. monocytogenes can persist throughout the salmon production system, but also showed that L. monocytogenes may be transmitted between slaughter and smoking operations or may be unique to smoking operations. While the majority of subtypes isolated have been rarely or never linked to human listeriosis cases, some subtypes have previously caused human listeriosis outbreaks and cases. Molecular subtyping thus is critical to identify L. monocytogenes transmission and niches in order to allow design and implementation of control strategies at the appropriate stage of production and in order to reduce the prevalence of L. monocytogenes linked to human disease.

Cloning of genes required for hypersensitivity and pathogenicity in Pseudomonas syringae pv. aptata.

PubMed

Minardi, P

1995-01-01

A genomic library of Pseudomonas syringae pv. aptata strain NCPPB 2664, which causes bacterial blight of sugar beet, lettuce and other plants, was constructed in the cosmid vector pCPP31. The 13.4 kb EcoRI fragment of the cosmid pHIR11, containing the hrp (hypersensitive response and pathogenicity) gene cluster of the closely related bacterium Pseudomonas syringae pv. syringae strain 61, was used as a probe to identify a homologous hrp gene cluster in P. syringae pv. aptata. Thirty of 2500 cosmid clones, screened by colony hybridization, gave a strong hybridization signal with the probe, but none of these conferred to the non-pathogenic bacterium, Pseudomonas fluorescens, the ability to elicit the hypersensitive response (HR) in tobacco. Southern blot analysis of EcoRI-digested genomic DNA of P. syringae pv. aptata showed hybridizing bands of 12 kb and 4.4 kb. Only a 12 kb fragment hybridized in digests of the cosmids. Cosmid clone pCPP1069 was mutagenized with Tn10-minitet and marker-exchanged into the genome of P. syringae pv. aptata. Three resulting prototrophic mutant strains failed to elicit the HR in tobacco and to cause disease in lettuce. The DNA flanking the Tn10-minitet insertions from mutated derivatives of pCPP1069 hybridized with the 10.6 kb Bg/II fragment of pHIR11. These results indicate that P. syringae pv. aptata harbours hrp genes that are similar to, but arranged differently from, homologous hrp genes of P. syringae pv. syringae.

Genotyping and chlorine-resistance of Methylobacterium aquaticum isolated from water samples in Japan.

PubMed

Furuhata, Katsunori; Banzai, Azusa U; Kawakami, Yasushi; Ishizaki, Naoto; Yoshida, Yoshihiro; Goto, Keiichi; Fukuyama, Masafumi

2011-09-01

For microbial ecological analysis, 14 strains of Methylobacterium aquaticum isolated from water samples were subjected to clustering analysis on the basis of ribotyping and RAPD-PCR tests. The ribopatterns after digestion with EcoRI obtained from 14 strains of M. aquaticum were used to divide the strains into two groups (Groups I and II) with a similarity of 55%. From the analysis of RAPD patterns using primer 208, the 14 strains were divided into 3 groups (A-C) based on a homology of 45% or greater, and from that using primer 272, there were 4 groups (A-D) based on a homology of 50% or greater. The chlorine resistance (99.9% CT values) of these isolates was also experimentally confirmed, and we attempted to define the connection between chlorine resistance and the geno-cluster. The average CT value of group I was 0.89 mg•min/l and the average of group II was 0.69 mg•min/l. No remarkable differences in the CT values for the groups were found.

Molecular Characterization of Plasmid-Mediated Oxytetracycline Resistance in Aeromonas salmonicida

PubMed Central

Adams, C. A.; Austin, B.; Meaden, P. G.; McIntosh, D.

1998-01-01

Using broth conjugation, we found that 19 of 29 (66%) oxytetracycline (OT)-resistant isolates of Aeromonas salmonicida transferred the OT resistance phenotype to Escherichia coli. The OT resistance phenotype was encoded by high-molecular-weight R-plasmids that were capable of transferring OT resistance to both environmental and clinical isolates of Aeromonas spp. The molecular basis for antibiotic resistance in OT-resistant isolates of A. salmonicida was determined. The OT resistance determinant from one plasmid (pASOT) of A. salmonicida was cloned and used in Southern blotting and hybridization experiments as a probe. The determinant was identified on a 5.4-kb EcoRI fragment on R-plasmids from the 19 OT-resistant isolates of A. salmonicida. Hybridization with plasmids encoding the five classes (classes A to E) of OT resistance determinants demonstrated that the OT resistance plasmids of the 19 A. salmonicida isolates carried the class A resistance determinant. Analysis of data generated from restriction enzyme digests showed that the OT resistance plasmids were not identical; three profiles were characterized, two of which showed a high degree of homology. PMID:9797265

Cloning of Pf3, a filamentous bacteriophage of Pseudomonas aeruginosa, into the pBD214 vector of Bacillus subtilis.

PubMed Central

Putterman, D G; Gryczan, T J; Dubnau, D; Day, L A

1983-01-01

The genome of Pf3, a filamentous single-stranded DNA bacteriophage of Pseudomonas aeruginosa (a gram-negative organism) was cloned into pBD214, a plasmid cloning vector of Bacillus subtilis (a gram-positive organism). Cloning in the gram-positive organism was done to avoid anticipated lethal effects. The entire Pf3 genome was inserted in each orientation at a unique Bc/I site within a thymidylate synthetase gene (from B. subtilis phage beta 22) on the plasmid. Additional clones were made by inserting EcoRI fragments of Pf3 DNA into a unique EcoRI site within this gene. Images PMID:6306273

Capture of unstable protein complex on the streptavidin-coated single-walled carbon nanotubes

NASA Astrophysics Data System (ADS)

Liu, Zunfeng; Voskamp, Patrick; Zhang, Yue; Chu, Fuqiang; Abrahams, Jan Pieter

2013-04-01

Purification of unstable protein complexes is a bottleneck for investigation of their 3D structure and in protein-protein interaction studies. In this paper, we demonstrate that streptavidin-coated single-walled carbon nanotubes (Strep•SWNT) can be used to capture the biotinylated DNA- EcoRI complexes on a 2D surface and in solution using atomic force microscopy and electrophoresis analysis, respectively. The restriction enzyme EcoRI forms unstable complexes with DNA in the absence of Mg2+. Capturing the EcoRI-DNA complexes on the Strep•SWNT succeeded in the absence of Mg2+, demonstrating that the Strep•SWNT can be used for purifying unstable protein complexes.

Variation in Ribosomal DNA among Isolates of the Mycorrhizal Fungus Cenococcum Geophilum FR.

NASA Astrophysics Data System (ADS)

Lobuglio, Katherine Frances

1990-01-01

Cenococcum geophilum Fr., a cosmopolitan mycorrhizal fungus, is well-known for its extremely wide host and habitat range. The ecological diversity of C. geophilum sharply contrasts its present taxonomic status as a monotypic form -genus. Restriction fragment length polymorphisms (RFLPs) in nuclear ribosomal DNA (rDNA) was used to assess the degree of genetic variation among 72 isolates of C. geophilum. The probe used in this study was the rDNA repeat cloned from C. geophilum isolate A145 (pCG15). Length of the rDNA repeat was approximately 9 kb. The rDNA clone was mapped for 5 restriction endonucleases. Hybridization with cloned Saccharomyces cerevisiae rDNA (pSR118, and pSR125 containing the 18S, and 5.8-25S rRNA genes respectively), and alignment of restriction endonuclease sites conserved in the rDNA genes of other fungi, were used to position the corresponding rDNAs of C. geophilum. Southern hybridizations with EcoRI, HindIII, XhoI, and PstI digested DNAs indicated extensive variation among the C. geophilum isolates, greater than has been previously reported to occur within a fungal species. Most of the rDNA polymorphisms occurred in the IGS region. Restriction endonuclease site and length polymorphisms were also observed in the 5.8S-26S genic regions. Sixteen size categories of length mutations, 6 restriction endonuclease site additions, and 4 restriction endonuclease site deletions were determined using isolate A145 as a reference. The rDNA repeat length among the isolates varied from approximately 8.5 to 10.2 kb. RFLPs were also observed in the mitochondrial (mt) 24S rRNA gene and flanking regions of HindIII digested DNAs of C. geophilum isolates representing both geographically distinct and similar origins. Among the C. geophilum isolates analyzed there were fewer RFLPs in mt-DNA than in nuclear rDNA. EcoRI rDNA phenotypes between C. geophilum and Elaphomyces anthracinus, its proposed teleomorph or sexual state, did not correspond. In addition, the four Elaphomyces species examined had smaller rDNA repeat sizes (approximately 7.3 to 8.0 kb) than that observed among C. geophilum isolates. UPGMA cluster analysis grouped C. geophilum isolates, on the basis of shared nuclear rDNA phenotypes, into a broad range of clusters ranging from 100% to 44% similarity. Limited correlation was observed among nuclear rDNA phenotypes and culture morphology, mycorrhizal characteristics, or geographic origins of the isolates. The amount of genetic variation demonstrated in this study indicates that C. geophilum is either an extremely heterogenous species or it represents more than one species and possibly more than one genus.

Morphological, chemical and molecular differentiation of Fusarium equiseti isolated from Norwegian cereals.

PubMed

Kosiak, Elzbieta Barbara; Holst-Jensen, Arne; Rundberget, Thomas; Gonzalez Jaen, Maria Teresa; Torp, Mona

2005-03-15

The morphological variation, secondary metabolite profiles and restriction fragment length polymorphisms (RFLPs) of PCR amplified intergenic spacer (IGS) ribosomal DNA (rDNA) were studied in 27 isolates of Fusarium equiseti, 25 isolated from Norwegian cereals and 2 from soil obtained from the IBT culture collection (BioCentrum, Technical University of Denmark). All 27 isolates were tested for production of fusarochromanone (FUSCHR), zearalenone (ZEA) and the trichothecenes: 15-monoacetoxy-scirpentriol (MAS), diacetoxy-scirpenol (DAS), T-2 and HT-2 toxins, T2-triol, neosolaniol (NEO), deoxynivalenol (DON), nivalenol (NIV) and 4-acetylnivalenol (Fus-X). The trichothecenes were analysed by GC-MS in a selected ion monitoring mode, while FUSCHR was determined by ion pair HPLC with fluorometric detection and production of ZEA by TLC. For amplification of IGS rDNA primers CNL12 and CNS1 were applied. IGS rDNA was digested with the four restriction enzymes: AvaII, CfoI, EcoRI and Sau3A. In addition, we sequenced the IGS rDNA region of three of the Norwegian isolates. There were two morphological types among the Norwegian strains of F. equiseti, type I with short apical cells (dominating) and type II with long apical cells, with four haplotypes identified based on the RFLP data. Variation in secondary metabolite profiles within and between the morphological groups was observed and the levels of produced toxins were: FUSCHR 3000-42,500 and 25-30 ng/g, NIV 20-2500 and 120-700 ng/g, FUS-X 20-15,000 and 0 ng/g, DAS 30-7500 and 0-600 ng/g, and MAS 10-600 and 0-500 ng/g, for strains with short and long apical cells, respectively. NEO was detected in 16/27 strains tested (all morphotype I). All but four strains of type I (these four lacked a restriction site for EcoRI) had identical RFLP profiles. The isolates of type II had two haplotypes. The IGS sequence similarity data indicated differences between these morphotypes corresponding to two separate lineages apparently at the species level.

Linkage map of the fragments of herpesvirus papio DNA.

PubMed Central

Lee, Y S; Tanaka, A; Lau, R Y; Nonoyama, M; Rabin, H

1981-01-01

Herpesvirus papio (HVP), an Epstein-Barr-like virus, causes lymphoblastoid disease in baboons. The physical map of HVP DNA was constructed for the fragments produced by cleavage of HVP DNA with restriction endonucleases EcoRI, HindIII, SalI, and PvuI, which produced 12, 12, 10, and 4 fragments, respectively. The total molecular size of HVP DNA was calculated as close to 110 megadaltons. The following methods were used for construction of the map; (i) fragments near the ends of HVP DNA were identified by treating viral DNA with lambda exonuclease before restriction enzyme digestion; (ii) fragments containing nucleotide sequences in common with fragments from the second enzyme digest of HVP DNA were examined by Southern blot hybridization; and (iii) the location of some fragments was determined by isolating individual fragments from agarose gels and redigesting the isolated fragments with a second restriction enzyme. Terminal heterogeneity and internal repeats were found to be unique features of HVP DNA molecule. One to five repeats of 0.8 megadaltons were found at both terminal ends. Although the repeats of both ends shared a certain degree of homology, it was not determined whether they were identical repeats. The internal repeat sequence of HVP DNA was found in the EcoRI-C region, which extended from 8.4 to 23 megadaltons from the left end of the molecule. The average number of the repeats was calculated to be seven, and the molecular size was determined to be 1.8 megadaltons. Similar unique features have been reported in EBV DNA (D. Given and E. Kieff, J. Virol. 28:524-542, 1978). Images PMID:6261015

Cloning of a Recombinant Plasmid Encoding Thiol-Specific Antioxidant Antigen (TSA) Gene of Leishmania majorand Expression in the Chinese Hamster Ovary Cell Line.

PubMed

Fatemeh, Ghaffarifar; Fatemeh, Tabatabaie; Zohreh, Sharifi; Abdolhosein, Dalimiasl; Mohammad Zahir, Hassan; Mehdi, Mahdavi

2012-01-01

TSA (thiol-specific antioxidant antigen) is the immune-dominant antigen of Leishmania major and is considered to be the most promising candidate molecule for a recombinant or DNA vaccine against leishmaniasis. The aim of the present work was to express a plasmid containing the TSA gene in eukaryotic cells. Genomic DNA was extracted, and the TSA gene was amplified by polymerase chain reaction (PCR). The PCR product was cloned into the pTZ57R/T vector, followed by subcloning into the eukaryotic expression vector pcDNA3 (EcoRI and HindIII sites). The recombinant plasmid was characterised by restriction digest and PCR. Eukaryotic Chinese hamster ovary cells were transfected with the plasmid containing the TSA gene. Expression of the L. major TSA gene was confirmed by sodium dodecyl sulphate-polyacrylamide gel electrophoresis and Western blotting. The plasmid containing the TSA gene was successfully expressed, as demonstrated by a band of 22.1 kDa on Western blots. The plasmid containing the TSA gene can be expressed in a eukaryotic cell line. Thus, the recombinant plasmid may potentially be used as a DNA vaccine in animal models.

A ZFY-like sequence in fish, with comments on the evolution of the ZFY family of genes in vertebrates

DOE Office of Scientific and Technical Information (OSTI.GOV)

Zimmerer, E.J.; Threlkeld, L.

1995-08-01

ZFY-like genes have been observed in a variety of vertebrate species. Although originally implicated as the primary testis-determining gene in humans and other placental mammals, more recent evidence indicates a role(s) outside that of testis determination. In this study, DNA from five species of fish, Carasius auratus, Rivulus marmoratus, Xiphophorus maculatus, X. milleri, and X. nigrensis was subjected to Southern blot analysis using a PCR-amplified fragment of mouse ZFY-like sequence as a probe. Restriction fragment patterns were not polymorphic between sexes in any one species but showed a different pattern for each species. With one exception, Rivulus, a 3.1-kb bandmore » from the EcoRI digestion was common to all. Sequence and open reading frame analysis of this fragment showed a strong homology to other known vertebrate ZFY-like genes. Of particular interest in this gene is a novel third finger domain similar to one human and one alligator ZFY-like gene. Our studies and others provide evidence for a family of vertebrate ZFY genes, with those having this novel third finger being representative of the ancestral condition. 30 refs., 3 figs., 3 tabs.« less

Identification and nucleotide sequence analysis of the repetitive DNA element in the genome of fish lymphocystis disease virus.

PubMed

Schnitzler, P; Delius, H; Scholz, J; Touray, M; Orth, E; Darai, G

1987-12-01

The genome of the fish lymphocystis disease virus (FLDV) was screened for the existence of repetitive DNA sequences using a defined and complete gene library of the viral genome (98 kbp) by DNA-DNA hybridization, heteroduplex analysis, and restriction fine mapping. A repetitive DNA sequence was detected at the coordinates 0.034 to 0.057 and 0.718 to 0.736 map units (m.u.) of the FLDV genome. The first region (0.034 to 0.057 m.u.) corresponds to the 5' terminus of the EcoRI FLDV DNA fragment B (0.034 to 0.165 m.u.) and the second region (0.718 to 0.736 m.u.) is identical to the EcoRI DNA fragment M of the viral genome. The DNA nucleotide sequence of the EcoRI FLDV DNA fragment M was determined. This analysis revealed the presence of many short direct and inverted repetitions, e.g., a 18-mer direct repetition (TTTAAAATTTAATTAA) that started at nucleotide positions 812 and 942 and a 14-mer inverted repeat (TTAAATTTAAATTT) at nucleotide positions 820 and 959. Only short open reading frames were detected within this region. The DNA repetitions are discussed as sequences that play a possible regulatory role for virus replication. Furthermore, hybridization experiments revealed that the repetitive DNA sequences are conserved in the genome of different strains of fish lymphocystis disease virus isolated from two species of Pleuronectidae (flounder and dab).

Dissemination of plasmid-encoded AmpC β-lactamases in antimicrobial resistant Salmonella serotypes originating from humans, pigs and the swine environment.

PubMed

Keelara, Shivaramu; Thakur, Siddhartha

2014-09-17

The aim of this study was to characterize and determine the inter-serovar exchange of AmpC β-lactamase conferring plasmids isolated from humans, pigs and the swine environment. Plasmids isolated from a total of 21 antimicrobial resistant (AMR) Salmonella isolates representing human clinical cases (n=6), pigs (n=6) and the swine farm environment (n=9) were characterized by replicon typing and restriction digestion, inter-serovar transferability by conjugation, and presence of AmpC β-lactamase enzyme encoding gene blaCMY-2 by southern hybridization. Based on replicon typing, the majority (17/21, 81%) of the plasmids belonged to the I1-Iγ Inc group and were between 70 and 103kb. The potential for inter-serovar plasmid transfer was further confirmed by the PCR detection of AMR genes on the plasmids isolated from trans-conjugants. Plasmids from Salmonella serovars Anatum, Ouakam, Johannesburg and Typhimurium isolated from the same cohort of pigs and their environment and S. Heidelberg from a single human clinical isolate had identical plasmids based on digestion with multiple restriction enzymes (EcoRI, HindIII and PstI) and southern blotting. We demonstrated likely horizontal inter-serovar exchange of plasmid-encoding AmpC β-lactamases resistance among MDR Salmonella serotypes isolated from pigs, swine farm environment and clinical human cases. This study provides valuable information on the role of the swine farm environment and by extension other livestock farm environments, as a potential reservoir of resistant bacterial strains that potentially transmit resistance determinants to livestock, in this case, swine, humans and possibly other hosts by horizontal exchange of plasmids. Copyright © 2014 Elsevier B.V. All rights reserved.

Characterization of class II alpha genes and DLA-D region allelic associations in the dog.

PubMed

Sarmiento, U M; Storb, R F

1988-10-01

Human major histocompatibility complex (HLA) cDNA probes were used to analyze the restriction fragment length polymorphism (RFLP) of the alpha genes of the DLA-D region in dogs. Genomic DNA from peripheral blood leucocytes of 23 unrelated DLA-D homozygous dogs representing nine DLA-D types (defined by mixed leucocyte reaction) was digested with restriction enzymes (BamHI, EcoRI, Hind III, Pvu II, Taq I, Rsa I, Msp I, Pst I and Bgl II), separated by agarose gel electrophoresis and transferred onto Biotrace membrane. The Southern blots were successively hybridized with radiolabelled HLA cDNA probes corresponding to DQ, DP, DZ and DR alpha genes. Clear evidence was obtained for the canine homologues of DQ and DR alpha genes with simple bi- or tri-allelic polymorphism respectively. Evidence for a single, nonpolymorphic DP alpha gene was also obtained. However, the presence of a DZ alpha gene could not be clearly demonstrated in canine genomic DNA. This report extends our previous RFLP analysis documenting polymorphism of DLA class II beta genes in the same panel of homozygous typing cell dogs, and provides the basis for DLA-D genotyping at a population level. This study also characterizes the RFLP-defined preferential allelic associations across the DLA-D region in nine different homozygous typing cell specificities.

Search for methylation-sensitive amplification polymorphisms in mutant figs.

PubMed

Rodrigues, M G F; Martins, A B G; Bertoni, B W; Figueira, A; Giuliatti, S

2013-07-08

Fig (Ficus carica) breeding programs that use conventional approaches to develop new cultivars are rare, owing to limited genetic variability and the difficulty in obtaining plants via gamete fusion. Cytosine methylation in plants leads to gene repression, thereby affecting transcription without changing the DNA sequence. Previous studies using random amplification of polymorphic DNA and amplified fragment length polymorphism markers revealed no polymorphisms among select fig mutants that originated from gamma-irradiated buds. Therefore, we conducted methylation-sensitive amplified polymorphism analysis to verify the existence of variability due to epigenetic DNA methylation among these mutant selections compared to the main cultivar 'Roxo-de-Valinhos'. Samples of genomic DNA were double-digested with either HpaII (methylation sensitive) or MspI (methylation insensitive) and with EcoRI. Fourteen primer combinations were tested, and on an average, non-methylated CCGG, symmetrically methylated CmCGG, and hemimethylated hmCCGG sites accounted for 87.9, 10.1, and 2.0%, respectively. MSAP analysis was effective in detecting differentially methylated sites in the genomic DNA of fig mutants, and methylation may be responsible for the phenotypic variation between treatments. Further analyses such as polymorphic DNA sequencing are necessary to validate these differences, standardize the regions of methylation, and analyze reads using bioinformatic tools.

Engineered selective plant male sterility through pollen-specific expression of the EcoRI restriction endonuclease.

PubMed

Millwood, Reginald J; Moon, Hong S; Poovaiah, Charleson R; Muthukumar, Balasubramaniam; Rice, John Hollis; Abercrombie, Jason M; Abercrombie, Laura L; Green, William Derek; Stewart, Charles Neal

2016-05-01

Unintended gene flow from transgenic plants via pollen, seed and vegetative propagation is a regulatory concern because of potential admixture in food and crop systems, as well as hybridization and introgression to wild and weedy relatives. Bioconfinement of transgenic pollen would help address some of these concerns and enable transgenic plant production for several crops where gene flow is an issue. Here, we demonstrate the expression of the restriction endonuclease EcoRI under the control of the tomato pollen-specific LAT52 promoter is an effective method for generating selective male sterility in Nicotiana tabacum (tobacco). Of nine transgenic events recovered, four events had very high bioconfinement with tightly controlled EcoRI expression in pollen and negligible-to-no expression other plant tissues. Transgenic plants had normal morphology wherein vegetative growth and reproductivity were similar to nontransgenic controls. In glasshouse experiments, transgenic lines were hand-crossed to both male-sterile and emasculated nontransgenic tobacco varieties. Progeny analysis of 16 000-40 000 seeds per transgenic line demonstrated five lines approached (>99.7%) or attained 100% bioconfinement for one or more generations. Bioconfinement was again demonstrated at or near 100% under field conditions where four transgenic lines were grown in close proximity to male-sterile tobacco, and 900-2100 seeds per male-sterile line were analysed for transgenes. Based upon these results, we conclude EcoRI-driven selective male sterility holds practical potential as a safe and reliable transgene bioconfinement strategy. Given the mechanism of male sterility, this method could be applicable to any plant species. © 2015 Society for Experimental Biology, Association of Applied Biologists and John Wiley & Sons Ltd.

Identification and properties of the largest subunit of the DNA-dependent RNA polymerase of fish lymphocystis disease virus: dramatic difference in the domain organization in the family Iridoviridae.

PubMed

Müller, M; Schnitzler, P; Koonin, E V; Darai, G

1995-05-01

Cytoplasmic DNA viruses encode a DNA-dependent RNA polymerase (DdRP) that is essential for transcription of viral genes. The amino acid sequences of the known largest subunits of DdRPs from different species contain highly conserved regions. Oligonucleotide primers, deduced from two conserved domains (RQP[T/S]LH and NADFDGDE) were used for detecting the corresponding gene of fish lymphocystis disease virus (FLCDV), a member of the family Iridoviridae, which replicates in the cytoplasm of infected cells of flatfish. The gene coding for the largest subunit of the DdRP was identified using a PCR-derived probe. The screening of the complete EcoRI gene library of the viral genome led to the identification of the gene locus of the largest subunit of the DdRP within the EcoRI DNA fragment B (12.4 kbp, 0.034 to 0.165 map units). The nucleotide sequence of a part (8334 bp) of the EcoRI DNA fragment B was determined and a large ORF on the lower strand (ATG = 5787; TAA = 2190) was detected which encodes a protein of 1199 amino acids. Comparison of the amino acid sequences of the largest subunits of the DdRP (RPO1) of FLCDV and Chilo iridescent virus (CIV) revealed a dramatic difference in their domain organization. Unlike the 1051 aa RPO1 of CIV, which lacks the C-terminal domain conserved in eukaryotic, eubacterial and other viral RNA polymerases, the 1199 aa RPO1 of FLCDV is fully collinear with its cellular and viral homologues. Despite this difference, comparative analysis of the amino acid sequences of viral and cellular RNA polymerases suggests a common origin for the largest RNA polymerase subunits of FLCDV and CIV.

The Influence of Primary and Secondary DNA Structure in Deletion and Duplication between Direct Repeats in Escherichia Coli

PubMed Central

Trinh, T. Q.; Sinden, R. R.

1993-01-01

We describe a system to measure the frequency of both deletions and duplications between direct repeats. Short 17- and 18-bp palindromic and nonpalindromic DNA sequences were cloned into the EcoRI site within the chloramphenicol acetyltransferase gene of plasmids pBR325 and pJT7. This creates an insert between direct repeated EcoRI sites and results in a chloramphenicol-sensitive phenotype. Selection for chloramphenicol resistance was utilized to select chloramphenicol resistant revertants that included those with precise deletion of the insert from plasmid pBR325 and duplication of the insert in plasmid pJT7. The frequency of deletion or duplication varied more than 500-fold depending on the sequence of the short sequence inserted into the EcoRI site. For the nonpalindromic inserts, multiple internal direct repeats and the length of the direct repeats appear to influence the frequency of deletion. Certain palindromic DNA sequences with the potential to form DNA hairpin structures that might stabilize the misalignment of direct repeats had a high frequency of deletion. Other DNA sequences with the potential to form structures that might destabilize misalignment of direct repeats had a very low frequency of deletion. Duplication mutations occurred at the highest frequency when the DNA between the direct repeats contained no direct or inverted repeats. The presence of inverted repeats dramatically reduced the frequency of duplications. The results support the slippage-misalignment model, suggesting that misalignment occurring during DNA replication leads to deletion and duplication mutations. The results also support the idea that the formation of DNA secondary structures during DNA replication can facilitate and direct specific mutagenic events. PMID:8325478

Highly repressible expression system for cloning genes that specify potentially toxic proteins.

PubMed Central

O'Connor, C D; Timmis, K N

1987-01-01

A highly repressible expression vector system that allows the cloning of potentially deleterious genes has been constructed. Undesired expression of a cloned gene was prevented (i) at the level of initiation of transcription, by the presence of the strong but highly repressible leftward promoter of bacteriophage lambda, lambda pL, and (ii) at the level of transcript elongation or translation, through synthesis of antisense RNA complementary to the mRNA of the cloned gene. The system was tested by measuring the inhibition of expression of traT, the gene for the TraT major outer membrane lipoprotein. Direct detection and functional assays indicated that an essentially complete inhibition of traT expression was obtained. As a further test of the system, the gene encoding the EcoRI restriction endonuclease was cloned in the absence of the gene of the corresponding protective EcoRI modification methylase. Transformants harboring this construct were only viable when both repression controls were operational. Images PMID:2443481

Molecular analysis of the anaerobic rumen fungus Orpinomyces - insights into an AT-rich genome.

PubMed

Nicholson, Matthew J; Theodorou, Michael K; Brookman, Jayne L

2005-01-01

The anaerobic gut fungi occupy a unique niche in the intestinal tract of large herbivorous animals and are thought to act as primary colonizers of plant material during digestion. They are the only known obligately anaerobic fungi but molecular analysis of this group has been hampered by difficulties in their culture and manipulation, and by their extremely high A+T nucleotide content. This study begins to answer some of the fundamental questions about the structure and organization of the anaerobic gut fungal genome. Directed plasmid libraries using genomic DNA digested with highly or moderately rich AT-specific restriction enzymes (VspI and EcoRI) were prepared from a polycentric Orpinomyces isolate. Clones were sequenced from these libraries and the breadth of genomic inserts, both genic and intergenic, was characterized. Genes encoding numerous functions not previously characterized for these fungi were identified, including cytoskeletal, secretory pathway and transporter genes. A peptidase gene with no introns and having sequence similarity to a gene encoding a bacterial peptidase was also identified, extending the range of metabolic enzymes resulting from apparent trans-kingdom transfer from bacteria to fungi, as previously characterized largely for genes encoding plant-degrading enzymes. This paper presents the first thorough analysis of the genic, intergenic and rDNA regions of a variety of genomic segments from an anaerobic gut fungus and provides observations on rules governing intron boundaries, the codon biases observed with different types of genes, and the sequence of only the second anaerobic gut fungal promoter reported. Large numbers of retrotransposon sequences of different types were found and the authors speculate on the possible consequences of any such transposon activity in the genome. The coding sequences identified included several orphan gene sequences, including one with regions strongly suggestive of structural proteins such as collagens and lampirin. This gene was present as a single copy in Orpinomyces, was expressed during vegetative growth and was also detected in genomes from another gut fungal genus, Neocallimastix.

Characterization of poxviruses from forest birds in Hawaii

USGS Publications Warehouse

Tripathy, Deoki N.; Schnitzlein, William M.; Morris, Patrick J.; Janssen, Don L.; Zuba, Jeffery K.; Massey, Greg; Atkinson, Carter T.

2000-01-01

Two strains of avian pox viruses were isolated from cutaneous lesions in Hawaiian crows (Corvus hawaiiensis) examined in 1994 and a third from a biopsy obtained in 1992 from an infected bird of the Apapane species (Himatione sanguinea) by inoculation of the chorioallantoic membranes (CAM) of developing chicken embryos. The resulting proliferative CAM lesions contained eosinophilic cytoplasmic inclusion bodies characteristic of pox virus infection. The pathogenicity of these three viruses in domestic chickens was mild as evidenced by the development of relatively minor lesions of short duration at the sites of inoculation. Their virulence in this host was similar to that of a fowlpox virus (FPV) vaccine strain and contrasted greatly with the ability of two field strains of FPV to produce extensive proliferative lesions. One of the Hawaiian crow pox virus isolates as well as the one originating from the Apapane species could be propagated in two secondary avian cell lines, QT-35 and LMH. A comparison of the restriction fragment length polymorphisms (RFLP) of the genomes of the two cell line-adapted viruses, generated by EcoRI digestion, revealed a limited degree of similarity. Moreover, neither profile was comparable to those of the two field isolates of FPV, which were almost indistinguishable from each other. Thus, based on the genetic distinctness of the two Hawaiian bird viruses, they appear to represent different strains of avipoxvirus.

Conservation of nodulation genes between Rhizobium meliloti and a slow-growing Rhizobium strain that nodulates a nonlegume host

PubMed Central

Marvel, Deborah J.; Kuldau, Gretchen; Hirsch, Ann; Richards, Eric; Torrey, John G.; Ausubel, Frederick M.

1985-01-01

Parasponia, a woody member of the elm family, is the only nonlegume genus whose members are known to form an effective nitrogen-fixing symbiosis with a Rhizobium species. The bacterial strain RP501 is a slow-growing strain of Rhizobium isolated from Parasponia nodules. Strain RP501 also nodulates the legumes siratro (Macroptilium atropurpureum) and cowpea (Vigna unguiculata). Using a cosmid clone bank of RP501 DNA, we isolated a 13.4-kilobase (kb) EcoRI fragment that complemented insertion and point mutations in three contiguous nodulation genes (nodABC) of Rhizobium meliloti, the endosymbiont of alfalfa (Medicago sativa). The complemented R. meliloti nod mutants induced effective nitrogen-fixing nodules on alfalfa seedlings but not on siratro, cowpeas, or Parasponia. The cloned RP501 nodulation locus hybridized to DNA fragments carrying the R. meliloti nodABC genes. A 3-kb cluster of Tn5 insertion mutations on the RP501 13.4-kb EcoRI fragment prevented complementation of R. meliloti nodABC mutations. Images PMID:16593600

Occurance of Staphylococcus nepalensis strains in different sources including human clinical material.

PubMed

Nováková, Dana; Pantůcek, Roman; Petrás, Petr; Koukalová, Dagmar; Sedlácek, Ivo

2006-10-01

Five isolates of coagulase-negative staphylococci were obtained from human urine, the gastrointestinal tract of squirrel monkeys, pig skin and from the environment. All key biochemical characteristics of the tested strains corresponded with the description of Staphylococcus xylosus species. However, partial 16S rRNA gene sequences obtained from analysed strains corresponded with those of Staphylococcus nepalensis reference strains, except for two strains which differed in one residue. Ribotyping with EcoRI and HindIII restriction enzymes, whole cell protein profile analysis performed by SDS-PAGE and SmaI macrorestriction analysis were used for more precise characterization and identification of the analysed strains. Obtained results showed that EcoRI and HindIII ribotyping and whole cell protein fingerprinting are suitable and reliable methods for the differentiation of S. nepalensis strains from the other novobiocin resistant staphylococci, whereas macrorestriction analysis was found to be a good tool for strain typing. The isolation of S. nepalensis is sporadic, and according to our best knowledge this study is the first report of the occurrence of this species in human clinical material as well as in other sources.

Deletion of the steroid-binding domain of the human androgen receptor gene in one family with complete androgen insensitivity syndrome: Evidence for further genetic heterogeneity in this syndrome

DOE Office of Scientific and Technical Information (OSTI.GOV)

Brown, T.R.; Lubahn, D.B.; Wilson, E.M.

1988-11-01

The cloning of a cDNA for the human androgen receptor gene has resulted in the availability for cDNA probes that span various parts of the gene, including the entire steroid-binding domain and part of the DNA-binding domain, as well as part of the 5' region of the gene. The radiolabeled probes were used to screen for androgen receptor mutations on Southern blots prepared by restriction endonuclease digestion of genomic DNA from human subjects with complete androgen insensitivity syndrome (AIS). In this investigation, the authors considered only patients presenting complete AIS and with the androgen receptor (-) form as the mostmore » probably subjects to show a gene deletion. One subject from each of six unrelated families with the receptor (-) form of complete AIS and 10 normal subjects were studied. In the 10 normal subjects and in 5 of the 6 patients, identical DNA restriction fragment patterns were observed with EcoRI and BamHI. Analysis of other members of this family confirmed the apparent gene deletion. The data provide direct proof that complete AIS in some families can result from a deletion of the androgen receptor structural gene. However, other families do not demonstrate such a deletion, suggesting that point mutations may also result in the receptor (-) form of complete AIS, adding further to the genetic heterogeneity of this syndrome.« less

Extraction of high-quality DNA from ethanol-preserved tropical plant tissues.

PubMed

Bressan, Eduardo A; Rossi, Mônica L; Gerald, Lee T S; Figueira, Antonio

2014-04-24

Proper conservation of plant samples, especially during remote field collection, is essential to assure quality of extracted DNA. Tropical plant species contain considerable amounts of secondary compounds, such as polysaccharides, phenols, and latex, which affect DNA quality during extraction. The suitability of ethanol (96% v/v) as a preservative solution prior to DNA extraction was evaluated using leaves of Jatropha curcas and other tropical species. Total DNA extracted from leaf samples stored in liquid nitrogen or ethanol from J. curcas and other tropical species (Theobroma cacao, Coffea arabica, Ricinus communis, Saccharum spp., and Solanum lycopersicon) was similar in quality, with high-molecular-weight DNA visualized by gel electrophoresis. DNA quality was confirmed by digestion with EcoRI or HindIII and by amplification of the ribosomal gene internal transcribed spacer region. Leaf tissue of J. curcas was analyzed by light and transmission electron microscopy before and after exposure to ethanol. Our results indicate that leaf samples can be successfully preserved in ethanol for long periods (30 days) as a viable method for fixation and conservation of DNA from leaves. The success of this technique is likely due to reduction or inactivation of secondary metabolites that could contaminate or degrade genomic DNA. Tissue conservation in 96% ethanol represents an attractive low-cost alternative to commonly used methods for preservation of samples for DNA extraction. This technique yields DNA of equivalent quality to that obtained from fresh or frozen tissue.

Extraction of high-quality DNA from ethanol-preserved tropical plant tissues

PubMed Central

2014-01-01

Background Proper conservation of plant samples, especially during remote field collection, is essential to assure quality of extracted DNA. Tropical plant species contain considerable amounts of secondary compounds, such as polysaccharides, phenols, and latex, which affect DNA quality during extraction. The suitability of ethanol (96% v/v) as a preservative solution prior to DNA extraction was evaluated using leaves of Jatropha curcas and other tropical species. Results Total DNA extracted from leaf samples stored in liquid nitrogen or ethanol from J. curcas and other tropical species (Theobroma cacao, Coffea arabica, Ricinus communis, Saccharum spp., and Solanum lycopersicon) was similar in quality, with high-molecular-weight DNA visualized by gel electrophoresis. DNA quality was confirmed by digestion with EcoRI or HindIII and by amplification of the ribosomal gene internal transcribed spacer region. Leaf tissue of J. curcas was analyzed by light and transmission electron microscopy before and after exposure to ethanol. Our results indicate that leaf samples can be successfully preserved in ethanol for long periods (30 days) as a viable method for fixation and conservation of DNA from leaves. The success of this technique is likely due to reduction or inactivation of secondary metabolites that could contaminate or degrade genomic DNA. Conclusions Tissue conservation in 96% ethanol represents an attractive low-cost alternative to commonly used methods for preservation of samples for DNA extraction. This technique yields DNA of equivalent quality to that obtained from fresh or frozen tissue. PMID:24761774

Physical mapping of the genomic DNA of the Oryctes rhinoceros baculovirus, KI.

PubMed

Mohan, K S; Gopinathan, K P

1991-11-15

A non-occluded baculovirus, OBV-KI has been isolated from the insect pest, Oryctes rhinoceros. The viral genome is estimated to be 123 kb, with a G + C content of 43 mol% and no detectible methylated bases. A restriction map of the OBV-KI genome for BamHI, EcoRI, HindIII, PstI, SalI and XbaI has been constructed.

Characterization of a New Type of Human Papillomavirus That Causes Skin Warts

PubMed Central

Orth, Gérard; Favre, Michel; Croissant, Odile

1977-01-01

A human papillomavirus (HPV) was isolated from the lesions of a patient (ML) bearing numerous hand common warts. This virus was compared with the well-characterized HPV found in typical plantar warts (plantar HPV). ML and plantar HPV DNAs have similar molecular weights (5.26 × 106 and 5.23 × 106, respectively) but were shown to be different by restriction enzyme analysis. When the cleavage products of both DNAs by endonuclease EcoRI, BamI, HpaI, or Hind were analyzed by electron microscopy, one, two, one, and four fragments were detected for ML HPV DNA instead of the two, one, two, and six fragments, respectively, detected for plantar HPV DNA. In contrast to plantar HPV DNA, a high proportion of ML HPV DNA molecules were resistant to these restriction enzymes. Most, if not all, of the molecules were either resistant to BamI and sensitive to EcoRI or sensitive to BamI and resistant to EcoRI. After denaturation and renaturation of the cleavage products of ML HPV DNA by a mixture of the two enzymes, the circular “heteroduplexes” formed showed one to three heterology loops corresponding to about 4 to 8% of the genome length. No sequence homology was detected between ML and plantar HPV DNAs by cRNA-DNA filter hybridization, by measuring the reassociation kinetics of an iodinated plantar HPV DNA in the presence of a 25-fold excess of ML HPV DNA, or by the heteroduplex technique. The two viruses had distinct electrophoretic polypeptide patterns and showed no antigenic cross-reaction by immunodiffusion or immunofluorescence techniques. Preliminary cRNA-DNA hybridization experiments, using viral DNAs from single or pooled plantar or hand warts, suggest that hand common warts are associated with viruses similar or related to ML HPV. The existence of at least two distinct types of HPVs that cause skin warts was demonstrated; they were provisionally called HPV type 1 and HPV type 2, with plantar HPV and ML HPV as prototypical viruses, respectively. Images PMID:198572

Variability and distribution of COL1A2 (type I collagen) polymorphisms in the central-eastern Mediterranean Basin.

PubMed

Scorrano, Gabriele; Lelli, Roberta; Martínez-Labarga, Cristina; Scano, Giuseppina; Contini, Irene; Hafez, Hani S; Rudan, Pavao; Rickards, Olga

2016-01-01

The most abundant of the collagen protein family, type I collagen is encoded by the COL1A2 gene. The COL1A2 restriction fragment length polymorphisms (RFLPs) EcoRI, RsaI and MspI in samples from several different central-eastern Mediterranean populations were analysed and found to be potentially informative anthropogenetic markers. The objective was to define the genetic variability of COL1A2 in the central-eastern Mediterranean and to shed light on its genetic distribution in human groups over a wide geographic area. PCR-RFLP analysis of EcoRI, RsaI and MspI polymorphisms of the COL1A2 gene was performed on oral swab and blood samples from 308 individuals from the central-eastern Mediterranean Basin. The genetic similarities among these groups and other populations described in the literature were investigated through correspondence analysis. Single-marker data and haplotype frequencies seemed to suggest a genetic homogeneity within the European populations, whereas a certain degree of differentiation was noted for the Egyptians and the Turks. The genetic variability in the central-eastern Mediterranean area is probably a result of the geographical barrier of the Mediterranean Sea, which separated European and African populations over time.

Label-free thioflavin T/G-quadruplex-based real-time strand displacement amplification for biosensing applications.

PubMed

Du, Yi-Chen; Zhu, Li-Na; Kong, De-Ming

2016-12-15

To promote application of strand-displacement amplification (SDA) techniques in biosensing, a label-free, real-time monitoring strategy for isothermal nucleic acid amplification reactions was designed. G-quadruplex structures were introduced into SDA products using specific recognition of G-quadruplexes by the fluorogenic dye thioflavin T. Performance was good for real-time monitoring of traditional SDA by a linear-amplification mechanism and for exponential cross-triggered SDA amplification. The strategy worked on a commercial real-time PCR instrument, making it suitable for biosensing platforms. As examples, two highly sensitive and specific biosensors were designed for analysis of the activity of uracil-DNA glycosylase (UDG) and the restriction endonuclease EcoRI. Detection limits were 6×10(-5)U/mL for UDG and 0.016U/mL for EcoRI. Detection of corresponding targets in complex matrices such as cell lysates or human serum was also demonstrated. Compared to traditional end-point detection methods, real-time SDA-based approaches have the advantages of simple, fast operation; high sensitivity; low risk of carryover contamination; and very high throughput. The introduction of real-time monitoring strategies may promote application of SDA reactions in biosensor design. Copyright © 2016 Elsevier B.V. All rights reserved.

Isolation and characterization of bacteriophages of Salmonella enterica serovar Pullorum.

PubMed

Bao, H; Zhang, H; Wang, R

2011-10-01

In this study, 2 bacteriophages of Salmonella Pullorum were isolated using an enrichment protocol and the double agar layer method. They were named PSPu-95 and PSPu-4-116, respectively, against clinical isolates of Salmonella Pullorum SPu-95 and SPu-116. The host ranges of the 2 bacteriophages were determined by performing spot tests with 20 bacteria strains. Both bacteriophages had wide host ranges. Bacteriophage PSPu-95 had a lytic effect on 17 of the 20 isolates (85%), and PSPu-4-116 produced a lytic effect on 14 isolates (70%) and was the only bacteriophage that produced a clear plaque on enterotoxigenic Escherichia coli K88. Transmission electron microscopy revealed the bacteriophages belonged to the order Caudovirales. Bacteriophage PSPu-95 was a member of the family Siphoviridae, but bacteriophage PSPu-4-116 belonged to the family Myoviridae. Both had a double-stranded DNA, which was digested with HindIII or EcoRI, that was estimated to be 58.3 kbp (PSPu-95) and 45.2 kbp (PSPu-4-116) by 1% agar electrophoresis. One-step growth kinetics showed that the latent periods were all less than 20 min, and the burst size was 77.5 pfu/cell for PSPu-95 and 86 pfu/cell for PSPu-4-116. The bacteriophages were able to survive in a pH range between 4 and 10, and they were able to survive in a treatment of 70°C for 60 min. The characterizations of these 2 bacteriophages were helpful in establishing a basis for adopting the most effective bacteriophage to control bacteria in the poultry industry.

In Vivo Imaging of mdrla Gene Expression

DTIC Science & Technology

2005-06-01

svImJ mouse strain, compatible with the ES cells used in our Transgenic Mouse Facility. b. Engineer PGK-neo and Renilla luciferase cassettes...inserted between the two loxP sites, upstream of the Neo cassette. A cloning strategy was then devised to fuse Renilla luciferase in-frame with the...sites: B, BamHI; E, EcoRI; S, ScaI. PGK-neo: neo under the control of the PGK promoter. Luc: Renilla luciferase fused in- frame with the translated

A molecular switch sensor for detection of PRSS1 genotype based on site-specific DNA cleavage of restriction endonuclease.

PubMed

Liu, Qicai; Gao, Feng; Weng, Shaohuang; Peng, Huaping; Lin, Liqing; Zhao, Chengfei; Lin, Xinhua

2015-01-01

PRSS1 mutations or polymorphism in the peripheral blood of patients can be used as susceptible molecular markers to pancreatic cancer. A sensor for selective electrochemical detection of PRSS1 genotypes was developed based on site-specific DNA cleavage of restriction endonuclease EcoRI. A mercapto-modified hairpin probe was immobilized on a gold electrode. The probe's neck can be cleaved by EcoRI in the absence of rs10273639 C/C of PRSS1 genotype, but it cannot be cleaved in the presence of T/T. The difference in quantity of electric charge was monitored by biosensors before and after enzymatic cleavage. Electrochemical signals are generated by differential pulse voltammetry interrogation of methylene blue (MB) that quantitatively binds to surface-confined hairpin probe via electrostatic interactions. The results suggested this method had a good specificity in distinguishing PRSS1 genotypes. There was a good linear relationship between the charge and the logarithmic function of PRSS1 rs10273639 T/T type DNA concentration (current=120.6303+8.8512log C, R=0.9942). The detection limit was estimated at 0.5 fM. The molecular switch sensor has several advantages, and it is possible to qualitatively, quantitatively, and noninvasively detect PRSS1 genotypes in the blood of patients with pancreatic cancer. © 2015 by the Association of Clinical Scientists, Inc.

Replication and Transcription of Eukaryotic DNA in Esherichia coli

PubMed Central

Morrow, John F.; Cohen, Stanley N.; Chang, Annie C. Y.; Boyer, Herbert W.; Goodman, Howard M.; Helling, Robert B.

1974-01-01

Fragments of amplified Xenopus laevis DNA, coding for 18S and 28S ribosomal RNA and generated by EcoRI restriction endonuclease, have been linked in vitro to the bacterial plasmid pSC101; and the recombinant molecular species have been introduced into E. coli by transformation. These recombinant plasmids, containing both eukaryotic and prokaryotic DNA, replicate stably in E. coli. RNA isolated from E. coli minicells harboring the plasmids hybridizes to amplified X. laevis rDNA. Images PMID:4600264

Translations on USSR Science and Technology, Biomedical and Behavioral Sciences, Number 33

DTIC Science & Technology

1978-07-05

7.6, 6.6 mM MgCl2, 6.6 mM 2-ß- mercaptoethanol, 2 y£ BamHI. Restriction was performed at 37°C for 15 min. We used, as marker DNA, EcoRI restrictor...linear DNA as function of molecular weight, plotted according to marker DNA to determine the molecular weights of DNA restrictors. Electron...according to the amp marker ; 3) reaction to agents that eliminate plasmids (acridine orange, ethidium bromide, sodium dodecylsulfate). The obtained

New genomic resources for switchgrass: a BAC library and comparative analysis of homoeologous genomic regions harboring bioenergy traits

PubMed Central

2011-01-01

Background Switchgrass, a C4 species and a warm-season grass native to the prairies of North America, has been targeted for development into an herbaceous biomass fuel crop. Genetic improvement of switchgrass feedstock traits through marker-assisted breeding and biotechnology approaches calls for genomic tools development. Establishment of integrated physical and genetic maps for switchgrass will accelerate mapping of value added traits useful to breeding programs and to isolate important target genes using map based cloning. The reported polyploidy series in switchgrass ranges from diploid (2X = 18) to duodecaploid (12X = 108). Like in other large, repeat-rich plant genomes, this genomic complexity will hinder whole genome sequencing efforts. An extensive physical map providing enough information to resolve the homoeologous genomes would provide the necessary framework for accurate assembly of the switchgrass genome. Results A switchgrass BAC library constructed by partial digestion of nuclear DNA with EcoRI contains 147,456 clones covering the effective genome approximately 10 times based on a genome size of 3.2 Gigabases (~1.6 Gb effective). Restriction digestion and PFGE analysis of 234 randomly chosen BACs indicated that 95% of the clones contained inserts, ranging from 60 to 180 kb with an average of 120 kb. Comparative sequence analysis of two homoeologous genomic regions harboring orthologs of the rice OsBRI1 locus, a low-copy gene encoding a putative protein kinase and associated with biomass, revealed that orthologous clones from homoeologous chromosomes can be unambiguously distinguished from each other and correctly assembled to respective fingerprint contigs. Thus, the data obtained not only provide genomic resources for further analysis of switchgrass genome, but also improve efforts for an accurate genome sequencing strategy. Conclusions The construction of the first switchgrass BAC library and comparative analysis of homoeologous harboring OsBRI1 orthologs present a glimpse into the switchgrass genome structure and complexity. Data obtained demonstrate the feasibility of using HICF fingerprinting to resolve the homoeologous chromosomes of the two distinct genomes in switchgrass, providing a robust and accurate BAC-based physical platform for this species. The genomic resources and sequence data generated will lay the foundation for deciphering the switchgrass genome and lead the way for an accurate genome sequencing strategy. PMID:21767393

New genomic resources for switchgrass: a BAC library and comparative analysis of homoeologous genomic regions harboring bioenergy traits.

PubMed

Saski, Christopher A; Li, Zhigang; Feltus, Frank A; Luo, Hong

2011-07-18

Switchgrass, a C4 species and a warm-season grass native to the prairies of North America, has been targeted for development into an herbaceous biomass fuel crop. Genetic improvement of switchgrass feedstock traits through marker-assisted breeding and biotechnology approaches calls for genomic tools development. Establishment of integrated physical and genetic maps for switchgrass will accelerate mapping of value added traits useful to breeding programs and to isolate important target genes using map based cloning. The reported polyploidy series in switchgrass ranges from diploid (2X = 18) to duodecaploid (12X = 108). Like in other large, repeat-rich plant genomes, this genomic complexity will hinder whole genome sequencing efforts. An extensive physical map providing enough information to resolve the homoeologous genomes would provide the necessary framework for accurate assembly of the switchgrass genome. A switchgrass BAC library constructed by partial digestion of nuclear DNA with EcoRI contains 147,456 clones covering the effective genome approximately 10 times based on a genome size of 3.2 Gigabases (~1.6 Gb effective). Restriction digestion and PFGE analysis of 234 randomly chosen BACs indicated that 95% of the clones contained inserts, ranging from 60 to 180 kb with an average of 120 kb. Comparative sequence analysis of two homoeologous genomic regions harboring orthologs of the rice OsBRI1 locus, a low-copy gene encoding a putative protein kinase and associated with biomass, revealed that orthologous clones from homoeologous chromosomes can be unambiguously distinguished from each other and correctly assembled to respective fingerprint contigs. Thus, the data obtained not only provide genomic resources for further analysis of switchgrass genome, but also improve efforts for an accurate genome sequencing strategy. The construction of the first switchgrass BAC library and comparative analysis of homoeologous harboring OsBRI1 orthologs present a glimpse into the switchgrass genome structure and complexity. Data obtained demonstrate the feasibility of using HICF fingerprinting to resolve the homoeologous chromosomes of the two distinct genomes in switchgrass, providing a robust and accurate BAC-based physical platform for this species. The genomic resources and sequence data generated will lay the foundation for deciphering the switchgrass genome and lead the way for an accurate genome sequencing strategy.

Use of molecular markers to compare Fusarium verticillioides pathogenic strains isolated from plants and humans.

PubMed

Chang, S C; Macêdo, D P C; Souza-Motta, C M; Oliveira, N T

2013-08-12

Fusarium verticillioides is a pathogen of agriculturally important crops, especially maize. It is considered one of the most important pathogens responsible for fumonisin contamination of food products, which causes severe, chronic, and acute intoxication in humans and animals. Moreover, it is recognized as a cause of localized infections in immunocompetent patients and disseminated infections among severely immunosuppressed patients. Several molecular tools have been used to analyze the intraspecific variability of fungi. The objective of this study was to use molecular markers to compare pathogenic isolates of F. verticillioides and isolates of the same species obtained from clinical samples of patients with Fusarium mycoses. The molecular markers that we used were inter-simple sequence repeat markers (primers GTG5 and GACA4), intron splice site primer (primer EI1), random amplified polymorphic DNA marker (primer OPW-6), and restriction fragment length polymorphism-internal transcribed spacer (ITS) from rDNA. From the data obtained, clusters were generated based on the UPGMA clustering method. The amplification products obtained using primers ITS4 and ITS5 and loci ITS1-5.8-ITS2 of the rDNA yielded fragments of approximately 600 bp for all the isolates. Digestion of the ITS region fragment using restriction enzymes such as EcoRI, DraI, BshI, AluI, HaeIII, HinfI, MspI, and PstI did not permit differentiation among pathogenic and clinical isolates. The inter-simple sequence repeat, intron splice site primer, and random amplified polymorphic DNA markers presented high genetic homogeneity among clinical isolates in contrast to the high variability found among the phytopathogenic isolates of F. verticillioides.

Autonomous parvovirus LuIII encapsidates equal amounts of plus and minus DNA strands

DOE Office of Scientific and Technical Information (OSTI.GOV)

Bates, R.C.; Snyder, C.E.; Banerjee, P.T.

1984-02-01

Autonomous parvoviruses are thought to uniquely encapsidate single-stranded DNA of minus polarity. In contrast, the defective adeno-associated viruses separately encapsidate equal amounts of plus and minus DNA strands. The uniqueness of minus strand encapsidation is reexamined for the autonomous parvoviruses. Although it was found that Kilham rat virus and H-1 virus encapsidate varying but small amounts of complementary-strand DNA, it was unexpected to find that LuIII virus encapsidated equal amounts of plus and minus DNA. The extracted LuIII DNA possessed properties of double-stranded replicative-form DNA, including insensitivity to S1 endonuclease, cleavage by restriction enzymes, and conversion to unit-length, single-stranded DNAmore » when electrophoresed under denaturing conditions. However, the inability of this DNA to form single-stranded DNA circles when denatured and then renatured in the presence of formamide and the lack of double-stranded DNA circle formation after treatment with exonuclease III and reannealing shows a lack of sequence homology of the 3' and 5' termini of LuIII DNA, in contrast to adeno-associated virus DNA. Digestion of LuIII double-stranded DNA with EcoRI and HincII and separation of plus and minus DNA strands on composite agarose-acrylamide gels identified a heterogeneity present only in the plus DNA strand. These results suggest that strand specificity of viral DNA encapsidation is not a useful property for differentiation between the autonomous and defective parvoviruses. Furthermore, encapsidation by LuIII of equal amounts of complementary DNA strands in contrast to encapsidation of minus strands by H-1 virus, when propagated in the same host cell type, suggests that selection of strands for encapsidation is a virus-coded rather than host-controlled event.« less

Epidemiological Investigation of Vaginal Saccharomyces cerevisiae Isolates by a Genotypic Method

PubMed Central

McCullough, Michael J.; Clemons, Karl V.; Farina, Claudio; McCusker, John H.; Stevens, David A.

1998-01-01

Saccharomyces cerevisiae is a ubiquitous, ascomycetous yeast, and vaginitis caused by this organism has been reported only very rarely. The aim of the present investigation was to assess the epidemiological relatedness of a group of vaginal and commercial S. cerevisiae isolates by a previously reported genetic typing method, which divided the isolates into two broad groups with numerous subtypes. Nineteen S. cerevisiae isolates obtained from patients suffering from vaginitis and four isolates from commercial products in the same city were analyzed. The cellular DNA from each isolate was digested with the restriction endonuclease EcoRI, and restriction fragment length polymorphisms were generated by horizontal gel electrophoresis. The results showed that although vaginal isolates did not cluster in any particular genetic subtype, multiple patients were infected with indistinguishable strains (there were nine distinct strains among 23 isolates). For two of three patients, all three with two episodes of S. cerevisiae vaginitis, different strains were isolated during the recurrence of this disease. Three other patients with indistinguishable isolates were epidemiologically related in that two were practitioners in the same clinic and the third was a patient at this clinic. We also found that one commercial strain was indistinguishable from the strain isolated from three different women at the time that they were suffering from vaginitis. The findings of the present study suggest that some S. cerevisiae strains may possess properties permitting persistence in the human host. Furthermore, person-to-person contact and the proliferation of the use of S. cerevisiae as a health-food product, in home baking, and in home brewing may be a contributing factor in human colonization and infection with this organism. PMID:9466776

Characterization of early follicular cDNA library suggests evidence for genetic polymorphisms in the inbred strain C108 of Bombyx mori.

PubMed

Mills, D R; Goldsmith, M R

2000-04-01

Recent work towards the completion of a saturated molecular genetic linkage map for the lepidopteran silkworm, Bombyx mori (n = 28), has provided evidence for existing polymorphisms in the inbred strain C108. Two inbred parental strains, p50 and C108, were crossed to produce the F1 (P/C) hybrid offspring. The populations used in this project were comprised of a combination of 29 F2 (F1 x F1) and 31 reciprocal backcross (P/C x C/C, P/C x P/P) progeny. All restriction fragment length polymorphisms (RFLPs) for the initial analysis were hybridized with anonymous probes derived from a random early follicular cDNA (Rcf) library from Bombyx. A total of 19 Rcf probes were selected as showing scorable codominant polymorphic patterns when screened against F2 and backcross DNAs digested with the restriction enzymes EcoRI, HindIII, or PstI, and Southern blotted to nylon membranes for hybridization. Of the newly reported Rcf probes, 7 (37%) were characterized as producing 'simple' polymorphic patterns, while 12 (63%) were characterized as producing 'complex' polymorphic patterns. Further characterization of the complex patterns subdivided this group into two general classes: polymorphisms that contained an additional allele, and multiple bands that contained an easily scored two banded polymorphism. Because the extra allele class was limited to the (P/C x C/C) backcross progeny, it is suggested that the inbred parental strain C108 harbors polymorphic loci that are inherited in a simple Mendelian fashion. A genetic analysis discussing plausible origins and maintenance of these polymorphisms is presented.

Dysfunctional growth hormone receptor in a strain of sex-linked dwarf chicken: evidence for a mutation in the intracellular domain.

PubMed

Agarwal, S K; Cogburn, L A; Burnside, J

1994-09-01

The sex-linked dwarf (dwdw) chicken represents a valuable animal model for studying GH insensitivity and the consequence of mutations in the GH receptor (GHR) gene. We have recently reported undetectable hepatic GH-binding activity and an aberrantly sized transcript in a strain of dwdw chickens obtained from Arbor Acre Farms, Inc. (Glastonbury, CT, USA). Southern blot analysis of the chicken GHR (cGHR) gene revealed a restriction-fragment length polymorphism in HindIII and EcoRI digests of genomic DNA in this strain of dwdw chicken. In order to localize the molecular mutation, we analysed the gene structure and determined the complete sequence of the 3' untranslated region (3' UTR) of the normal cGHR. With the use of this information, we located a large deletion in the 3' end of the cGHR gene of the Connecticut (CT) strain of dwdw chicken. This deletion (1773 bp) contained 27 highly conserved amino acids of the 3' end of the coding region, the in-frame stop codon, a less frequently used poly(A) signal that is normally found 445 bp downstream of the stop codon, and a large portion of the 3' UTR. Because of this deletion, 27 novel amino acids were substituted and the open reading frame was extended for an additional 26 amino acids before reaching the transcriptional termination site. The predicted amino acid sequence of the novel carboxyl-terminus of the dwdw cGHR is largely hydrophobic with a polylysine tail, whereas the carboxyl-terminus of the wild-type (DwDw) cGHR is composed of hydrophilic amino acids.(ABSTRACT TRUNCATED AT 250 WORDS)

Detection of tetQ and ermF antibiotic resistance genes in Prevotella and Porphyromonas isolates from clinical specimens and resident microbiota of humans.

PubMed

Arzese, A R; Tomasetig, L; Botta, G A

2000-05-01

Gram-negative anaerobes belonging to the genera Fusobacterium, Prevotella and Porphyromonas were investigated for the presence of tetQ and ermF, which have been shown to be spread by conjugal elements. One hundred isolates from either sites of infection or various body sites in healthy subjects were studied. PCR was used to detect tetQ, and DNA-DNA hybridization studies on EcoRI chromosomal digests were undertaken to detect the presence of tetQ and ermF. Antibiotic sensitivity assays were performed on selected isolates to detect tetracycline, erythromycin and penicillin resistance. Twenty Fusobacterium isolates lacked tetQ, and were tetracycline sensitive. Twenty per cent of Prevotella spp. isolates both from clinical specimens and from healthy subjects were found to possess tetQ. Of 20 Porphyromonas isolates tested, one (Porphyromonas levii) from a case of bacterial vaginosis was shown to possess tetQ in the chromosome. The presence of tetQ was always associated with tetracycline resistance. Four isolates of Prevotella melaninogenica and one isolate of Prevotella were ermF-positive, although expression of erythromycin resistance was not consistently associated with detection of this gene. Antibiotic resistance phenotypes of Prevotella isolates were shown to be related to specific chromosomal restriction patterns by hybridization studies: tetracycline resistance and tetracycline/erythromycin resistance are conferred by Bacteroides tetracycline-resistant ERL elements, whereas the tetracycline/penicillin resistance phenotype could be due to spread of elements identified in Prevotella only. Tetracycline/erythromycin-resistant and tetracycline/erythromycin/penicillin-resistant P. melaninogenica isolates were found in this study. It appeared that the presence of tetQ and ermF in Bacteroides and Prevotella contributed to the persistence of antibiotic resistance isolates within the host and to potential spread to other organisms through conjugal elements.

Overexpression of Salmonella enterica serovar Typhi recA gene confers fluoroquinolone resistance in Escherichia coli DH5α.

PubMed

Yassien, M A M; Elfaky, M A

2015-11-01

A spontaneous fluoroquinolone-resistant mutant (STM1) was isolated from its parent Salmonella enterica serovar Typhi (S. Typhi) clinical isolate. Unlike its parent isolate, this mutant has selective resistance to fluoroquinolones without any change in its sensitivity to various other antibiotics. DNA gyrase assays revealed that the fluoroquinolone resistance phenotype of the STM1 mutant did not result from alteration of the fluoroquinolone sensitivity of the DNA gyrase isolated from it. To study the mechanism of fluoroquinolone resistance, a genomic library from the STM1 mutant was constructed in Escherichia coli DH5α and two recombinant plasmids were obtained. Only one of these plasmids (STM1-A) conferred the selective fluoroquinolone resistance phenotype to E. coli DH5α. The chromosomal insert from STM1-A, digested with EcoRI and HindIII restriction endonucleases, produced two DNA fragments and these were cloned separately into pUC19 thereby generating two new plasmids, STM1-A1 and STM1-A2. Only STM1-A1 conferred the selective fluoroquinolone resistance phenotype to E. coli DH5α. Sequence and subcloning analyses of STM1-A1 showed the presence of an intact RecA open reading frame. Unlike that of the wild-type E. coli DH5α, protein analysis of a crude STM1-A1 extract showed overexpression of a 40 kDa protein. Western blotting confirmed the 40 kDa protein band to be RecA. When a RecA PCR product was cloned into pGEM-T and introduced into E. coli DH5α, the STM1-A11 subclone retained fluoroquinolone resistance. These results suggest that overexpression of RecA causes selective fluoroquinolone resistance in E. coli DH5α.

An electrochemiluminescence biosensor for endonuclease EcoRI detection.

PubMed

Li, Yingjie; Li, Yuqin; Wu, Yaoyu; Lu, Fushen; Chen, Yaowen; Gao, Wenhua

2017-03-15

Endonucleases cleavage of DNA plays an important role in biological and medicinal chemistry. This work was going to develop a reliable and sensitive electrochemiluminescent (ECL) biosensor for detecting endonucleases by using gold nanoparticles graphene composite (GNPs-graphene) as a signal amplifier. Firstly, the GNPs and graphene were simultaneously deposited on the glassy carbon electrode (GCE) by cyclic voltammetry. Then a stem DNA was anchored on the surface of GCE. And with a modifying DNA introduced into the electrode by DNA assembly, a strong ECL signal was obtained. After a DNA modified with ferrocene assembly to the stem DNA, the ECL signal had a sharp decrease due to the quench effect of ferrocene to and the biosensor comes into being a "off" state. With the effect of endonuclease, the ECL signal had a recovery because of the ferrocene being released and the biosensor formed a "on" state. Moreover, the recovery of ECL signal was related to the concentration of endonucleases. Combining specific defined DNA and endonuclease, this method has a potential to detect different endonucleases. In this work, we took the EcoRI as an example to identify the feasibility of ECL biosensor in applying in sensitive detection of endonucleases using a GNPs-graphene signal amplifier. Under optimal condition, the proposed biosensor obtained a low limit of detection (LOD) 5.6×10 -5 UmL -1 . And the stability, selectivity and reproducibility of the biosensor also were researched. Copyright © 2016 Elsevier B.V. All rights reserved.

Frequencies of occurrence of all human chromosomes in micronuclei from normal and 5-azacytidine-treated lymphocytes as revealed by chromosome painting.

PubMed

Fauth, E; Scherthan, H; Zankl, H

1998-05-01

Chromosome painting with library DNA probes specific for all human chromosomes was used to study the chromosomal content of micronuclei (MN) in normal and 5-azacytidine (5-aza-C)-treated lymphocyte cultures. More than 60,000 normal lymphocytes were screened for associated MN after in situ hybridization. At least 50 MN were scored for each probe. With the exception of chromosomes 12 and 19, which did not occur in MN, all other chromosomes were detected in MN at frequencies varying from 1 to 11.5%. Treatment of lymphocyte cultures with 5-aza-C induced preferential exclusion of chromosomes 1 (34%), 9 (32%) and 16 (20%) material in MN, whereas chromosome 8, 10, 12-15 and 21 material was not detected in MN. The results obtained from normal lymphocytes allow for the first time an estimation of the frequency of occurrence of all chromosomes in spontaneously occurring MN in human cells. Data derived from 5-aza-C-treated lymphocytes are furthermore consistent with the view that undermethylation of heterochromatin may be associated with loss of specific chromosomes at metaphase.

Identification of Two Candidate Tumor Suppressor Genes on Chromosome 17p13.3: Assessment of Their Roles in Breast and Ovarian Carcinogenesis

DTIC Science & Technology

1997-07-01

minimum region of allelic loss on chromosome 17p 13.3, between polymorphic markers D17S5 and D17S28, in genomic DNA from breast and ovarian tumors (Figure 1...encode proteins of 443 and 227 amino acids, with no known functional motifs. Comparison of genomic and cDNA sequences showed that the genes overlap...is tissue specific (Figure 4). When zoo blots comprised of EcoRI fragments of genomic DNA from various species were probed with the unique exon 1 of

Biochemical and genetic analyses of acetoin catabolism in Alcaligenes eutrophus.

PubMed Central

Fründ, C; Priefert, H; Steinbüchel, A; Schlegel, H G

1989-01-01

In genetic studies on the catabolism of acetoin in Alcaligenes eutrophus, we used Tn5::mob-induced mutants which were impaired in the utilization of acetoin as the sole carbon source for growth. The transposon-harboring EcoRI restriction fragments from 17 acetoin-negative and slow-growing mutants (class 2a) and from six pleiotropic mutants of A. eutorphus, which were acetoin-negative and did not grow chemolithoautotrophically (class 2b), were cloned from pHC79 gene banks. The insertions of Tn5 were mapped on four different chromosomal EcoRI restriction fragments (A, C, D, and E) in class 2a mutants. The native DNA fragments were cloned from a lambda L47 or from a cosmid gene bank. Evidence is provided that fragments A (21 kilobase pairs [kb]) and C (7.7 kb) are closely linked in the genome; the insertions of Tn5 covered a region of approximately 5 kb. Physiological experiments revealed that this region encodes for acetoin:dichlorophenol-indophenol oxidoreductase, a fast-migrating protein, and probably for one additional protein that is as yet unknown. In mutants which were not completely impaired in growth on acetoin but which grew much slower and after a prolonged lag phase, fragments D (7.2 kb) and E (8.1 kb) were inactivated by insertion of Tn5::mob. No structural gene could be assigned to the D or E fragments. In class 2b mutants, insertions of Tn5 were mapped on fragment B (11.3 kb). This fragment complemented pleiotropic hno mutants in trans; these mutants were impaired in the formation of a rpoN-like protein. The expression of the gene cluster on fragments A and C seemed to be rpoN dependent. PMID:2556366

Chemoraces and Habitat Specialization of Claviceps purpurea Populations

PubMed Central

Pažoutová, Sylvie; Olšovská, Jana; Linka, Marek; Kolínská, Renata; Flieger, Miroslav

2000-01-01

We studied genetic variability of 100 isolates of Claviceps purpurea by using randomly amplified polymorphic DNA (RAPD), an EcoRI restriction site polymorphism in the 5.8S ribosomal DNA (rDNA), the alkaloids produced, and conidial morphology. We identified three groups: (i) group G1 from fields and open meadows (57 isolates), (ii) group G2 from shady or wet habitats (41 isolates), and (iii) group G3 from Spartina anglica from salt marshes (2 isolates). The sclerotia of G1 isolates contained ergotamines and ergotoxines; G2 isolates produced ergosine and ergocristine along with small amounts of ergocryptine; and G3 isolates produced ergocristine and ergocryptine. The conidia of G1 isolates were 5 to 8 μm long, the conidia of G2 isolates were 7 to 10 μm long, and the conidia of G3 isolates were 10 to 12 μm long. Sclerotia of the G2 and G3 isolates floated on water. In the 5.8S rDNA analysis, an EcoRI site was found in G1 and G3 isolates but not in G2 isolates. The host preferences of the groups were not absolute, and there were host genera that were common to both G1 and G2; the presence of members of different groups in the same locality was rare. Without the use of RAPD or rDNA polymorphism, it was not possible to distinguish the three groups solely on the basis of phenotype, host, or habitat. In general, populations of C. purpurea are not host specialized, as previously assumed, but they are habitat specialized, and collecting strategies and toxin risk assessments should be changed to reflect this paradigm shift. PMID:11097923

Use of a repetitive DNA probe to type clinical and environmental isolates of Aspergillus flavus from a cluster of cutaneous infections in a neonatal intensive care unit.

PubMed

James, M J; Lasker, B A; McNeil, M M; Shelton, M; Warnock, D W; Reiss, E

2000-10-01

Aspergillus flavus is second to A. fumigatus as a cause of invasive aspergillosis, but no standard method exists for molecular typing of strains from human sources. A repetitive DNA sequence cloned from A. flavus and subcloned into a pUC19 vector, pAF28, was used to type 18 isolates from diverse clinical, environmental, and geographic sources. The restriction fragment length polymorphisms generated with EcoRI- or PstI-digested genomic DNA and probed with digoxigenin-labeled pAF28 revealed complete concordance between patterns. Eighteen distinct fingerprints were observed. The probe was used to investigate two cases of cutaneous A. flavus infection in low-birth-weight infants in a neonatal intensive care unit (NICU). Both infants were transported by the same ambulance and crew to the NICU on the same day. A. flavus strains of the same genotype were isolated from both infants, from a roll of tape used to fasten their umbilical catheters, from a canvas bag used to store the tape in the ambulance, and from the tape tray in the ambulance isolette. These cases highlight the need to consider exposures in critically ill neonates that might occur during their transport to the NICU and for stringent infection control practices. The hybridization profiles of strains from a second cluster of invasive A. flavus infections in two pediatric hematology-oncology patients revealed a genotype common to strains from a definite case patient and a health care worker. A probable case patient was infected with a strain with a genotype different from that of the strain from the definite case patient but highly related to that of an environmental isolate. The high degree of discrimination and reproducibility obtained with the pAF28 probe underscores its utility for typing clinical and environmental isolates of A. flavus.

Use of a Repetitive DNA Probe To Type Clinical and Environmental Isolates of Aspergillus flavus from a Cluster of Cutaneous Infections in a Neonatal Intensive Care Unit

PubMed Central

James, Michael J.; Lasker, Brent A.; McNeil, Michael M.; Shelton, Mark; Warnock, David W.; Reiss, Errol

2000-01-01

Aspergillus flavus is second to A. fumigatus as a cause of invasive aspergillosis, but no standard method exists for molecular typing of strains from human sources. A repetitive DNA sequence cloned from A. flavus and subcloned into a pUC19 vector, pAF28, was used to type 18 isolates from diverse clinical, environmental, and geographic sources. The restriction fragment length polymorphisms generated with EcoRI- or PstI-digested genomic DNA and probed with digoxigenin-labeled pAF28 revealed complete concordance between patterns. Eighteen distinct fingerprints were observed. The probe was used to investigate two cases of cutaneous A. flavus infection in low-birth-weight infants in a neonatal intensive care unit (NICU). Both infants were transported by the same ambulance and crew to the NICU on the same day. A. flavus strains of the same genotype were isolated from both infants, from a roll of tape used to fasten their umbilical catheters, from a canvas bag used to store the tape in the ambulance, and from the tape tray in the ambulance isolette. These cases highlight the need to consider exposures in critically ill neonates that might occur during their transport to the NICU and for stringent infection control practices. The hybridization profiles of strains from a second cluster of invasive A. flavus infections in two pediatric hematology-oncology patients revealed a genotype common to strains from a definite case patient and a health care worker. A probable case patient was infected with a strain with a genotype different from that of the strain from the definite case patient but highly related to that of an environmental isolate. The high degree of discrimination and reproducibility obtained with the pAF28 probe underscores its utility for typing clinical and environmental isolates of A. flavus. PMID:11015372

Genetic differentiation in Pyrenophora teres f. teres populations from Syria and Tunisia as assessed by AFLP markers.

PubMed

Bouajila, A; Zoghlami, N; Murad, S; Baum, M; Ghorbel, A; Nazari, K

2013-06-01

To investigate the level of genetic differentiation and diversity among Pyrenophora teres isolate populations originating from different agro-ecological areas of Syria and Tunisia and to determine the potential of AFLP profiling in genotyping Pyrenophora teres f. teres. In this study, AFLP markers have been employed to identify patterns of population structure in 20 Pyrenophora teres f. teres populations from Syria and Tunisia. Ninety-four isolates were studied by the use of a protocol that involved stringent PCR amplification of fragments derived from digestion of genomic DNA with restriction enzymes EcoRI and MesI. Based on 401 amplified polymorphic DNA markers (AFLP), variance analyses indicated that most of the variation was partitioned within rather than between populations. Genotypic diversity (GD) was high for populations from Rihane, local landraces and different agro-ecological zones (GD = 0·75-0·86). There was high genetic differentiation among pathogen populations from different host populations in Syria (Gst  = 0·31, ht = 0·190) and Tunisia (Gst  = 0·39, ht = 0·263), which may be partly explained by the low gene flow around the areas sampled. A phenetic tree revealed three groups with high bootstrap values (55, 68, 76) and reflected the grouping of isolates based on host, or agro-ecological areas. AFLP profiling is an effective method for typing the genetically diverse pathogen Pyrenophora teres f. teres. The study represents a comparative analysis of the genetic diversity in P. teres isolates from two countries spanning two continents and also shows that several distinct P. teres genotypes may be found in a given environment. The implications of these findings for Pyrenophora teres f. teres evolutionary potential and net blotch-resistance breeding in Syria and Tunisia were also discussed. © 2012 The Society for Applied Microbiology.

Molecular analysis of DiGeorge Syndrome-related translocation breakpoints in 22q11.2

DOE Office of Scientific and Technical Information (OSTI.GOV)

Chieffo, C.; Barnoski, B.L.; Emanuel, B.S.

1994-09-01

22q11 demonstrates a high frequency of disease-specific rearrangements. Several of the rearrangements are associated with developmental abnormalities such as DiGeorge Syndrome (DGS), Velocardiofacial Syndrome (VCFS), Cat Eye Syndrome (CES) and Supernumerary der(22)t(11;22) Syndrome. DGS and VCFS involve deletions of 22q11.2 resulting from unbalanced translocations or microdeletions. In contrast, CES and Supernumerary der(22)t(11;22) Syndrome result from duplications of this region via inter- or intra- chromosomal exchange. Although the molecular mechanism giving rise to these rearrangements has yet to be elucidated, the presence of known 22q11 repetitive elements are likely to be involved. GM5878 is a 46,XY,t(10;22) cell line from a balancedmore » translocation carrier father of an unbalanced DGS patient. GM0980 is a cell line from a patient with features of DGS/VCFS with an unbalanced karyotype. Using FISH cosmids, we have localized these translocation breakpoints near pH160b (D22S66) which maps to the center of the DiGeorge chromosomal region (DGCR). To further localize the breakpoint of GM5878, overlapping cosmids spanning this region were used as probes for FISH. Use of additional overlapping cosmids allowed the sublocalization of the breakpoint to a 10kb region. A 4.8 kb BglII fragment predicted to cross the breakpoint was isolated. When this fragment was used as a probe to normal and GM5878 DNA, novel bands were detected in GM5878 DNA digested with EcoRI and BglII. Similar analysis of the GM0980 breakpoint is being pursued. Full molecular characterization of these translocations is in progress using inverse PCR to clone the junctional fragments for sequencing. Detailed analysis of the region may reveal molecular features which make this a rearrangement prone area of the genome and help elucidate its relationship to human cytogenetic disease.« less

Inhibition of Autoimmune Chagas-Like Heart Disease by Bone Marrow Transplantation

PubMed Central

Guimaro, Maria C.; Alves, Rozeneide M.; Rose, Ester; Sousa, Alessandro O.; de Cássia Rosa, Ana; Hecht, Mariana M.; Sousa, Marcelo V.; Andrade, Rafael R.; Vital, Tamires; Plachy, Jiří; Nitz, Nadjar; Hejnar, Jiří; Gomes, Clever C.; L. Teixeira, Antonio R.

2014-01-01

Background Infection with the protozoan Trypanosoma cruzi manifests in mammals as Chagas heart disease. The treatment available for chagasic cardiomyopathy is unsatisfactory. Methods/Principal Findings To study the disease pathology and its inhibition, we employed a syngeneic chicken model refractory to T. cruzi in which chickens hatched from T. cruzi inoculated eggs retained parasite kDNA (1.4 kb) minicircles. Southern blotting with EcoRI genomic DNA digests revealed main 18 and 20 kb bands by hybridization with a radiolabeled minicircle sequence. Breeding these chickens generated kDNA-mutated F1, F2, and F3 progeny. A targeted-primer TAIL-PCR (tpTAIL-PCR) technique was employed to detect the kDNA integrations. Histocompatible reporter heart grafts were used to detect ongoing inflammatory cardiomyopathy in kDNA-mutated chickens. Fluorochromes were used to label bone marrow CD3+, CD28+, and CD45+ precursors of the thymus-dependent CD8α+ and CD8β+ effector cells that expressed TCRγδ, vβ1 and vβ2 receptors, which infiltrated the adult hearts and the reporter heart grafts. Conclusions/Significance Genome modifications in kDNA-mutated chickens can be associated with disruption of immune tolerance to compatible heart grafts and with rejection of the adult host's heart and reporter graft, as well as tissue destruction by effector lymphocytes. Autoimmune heart rejection was largely observed in chickens with kDNA mutations in retrotransposons and in coding genes with roles in cell structure, metabolism, growth, and differentiation. Moreover, killing the sick kDNA-mutated bone marrow cells with cytostatic and anti-folate drugs and transplanting healthy marrow cells inhibited heart rejection. We report here for the first time that healthy bone marrow cells inhibited heart pathology in kDNA+ chickens and thus prevented the genetically driven clinical manifestations of the disease. PMID:25521296

Recombination hot spot in 3.2-kb region of the Charcot-Marie Tooth type 1A repeat sequences: New tools for molecular diagnosis of hereditary neuropathy with liability to pressure palsies and of Charcot-Marie-Tooth type 1A

DOE Office of Scientific and Technical Information (OSTI.GOV)

Lopes, J.; LeGuern, E.; Gouider, R.

1996-06-01

Charcot-Marie-Tooth type 1A (CMT1A) disease and hereditary neuropathy with liability to pressure palsies (HNPP) are autosomal dominant neuropathies, associated, respectively, with duplications and deletions of the same 1.5-Mb region on 17p11.2-p12. These two rearrangements are the reciprocal products of an unequal meiotic crossover between the two chromosome 17 homologues, caused by the misalignment of the CMT1A repeat sequences (CMT1A-REPs), the homologous sequences flanking the 1.5-Mb CMT1A/HNPP monomer unit. In order to map recombination breakpoints within the CMT1A-REPs, a 12.9-kb restriction map was constructed from cloned EcoRI fragments of the proximal and distal CMT1A-REPs. Only 3 of the 17 tested restrictionmore » sites were present in the proximal CMT1A-REP but absent in the distal CMT1A-REP, indicating a high degree of homology between these sequences. The rearrangements were mapped in four regions of the CMT1A-REPs by analysis of 76 CMT1A index cases and 38 HNPP patients, who were unrelated. A hot spot of crossover breakpoints located in a 3.2-kb region accounted for three-quarters of the rearrangements, detected after EcoRI/SacI digestion, by the presence of 3.2-kb and 7.8-kb junction fragments in CMT1A and HNPP patients, respectively. These junction fragments, which can be detected on classical Southern blots, permit molecular diagnosis. Other rearrangements can also be detected by gene dosage on the same Southern blots. 25 refs., 4 figs., 2 tabs.« less

A 1.5-Mb deletion in 17p11.2-p12 is frequently observed in Italian families with hereditary neuropathy with liability to pressure palsies

DOE Office of Scientific and Technical Information (OSTI.GOV)

Lorenzetti, D.; Pandolfo, M.; Pareyson, D.

1995-01-01

Hereditary neuropathy with liability to pressure palsies (HNPP) is an autosomal dominant disorder characterized by recurrent mononeuropathies. A 1.5-Mb deletion in chromosome 17p11.2-p12 has been associated with HNPP. Duplication of the same 1.5-Mb region is known to be associated with Charcot-Marie-Tooth disease type 1 (CMT1A), a more severe peripheral neuropathy characterized by symmetrically slowed nerve conduction velocity (NCV). The CMT1A duplication and HNPP deletion appear to be the reciprocal products of a recombination event involving a repeat element (CMT1A-REP) that flanks the 1.5-Mb region involved in the duplication/deletion. Patients from nine unrelated Italian families who were diagnosed with HNPP onmore » the basis of clinical, electrophysiological, and histological evaluations were analyzed by molecular methods for DNA deletion on chromosome 17p. In all nine families, Southern analysis using a CMT1A-REP probe detected a reduced hybridization signal of a 6.0-kb EcoRI fragment mapping within the distal CMT1A-REP, indicating deletion of one copy of CMT1A-REP in these HNPP patients. Families were also typed with a polymorphic (CA){sub n} repeat and with RFLPs corresponding to loci D17S122, D17S125, and D17S61, which all map within the deleted region. Lack of allelic transmission from affected parent to affected offspring was observed in four informative families, providing an independent indication for deletion. Furthermore, pulsed-field gel electrophoresis analysis of SacII-digested genomic DNA detected junction fragments specific to the 1.5-Mb HNPP deletion in seven of nine Italian families included in this study. These findings suggest that a 1.5-Mb deletion on 17p11.2-p12 is the most common mutation associated with HNPP. 51 refs., 5 figs., 1 tab.« less

Cloning of the citrate permease gene of Lactococcus lactis subsp. lactis biovar diacetylactis and expression in Escherichia coli.

PubMed Central

Sesma, F; Gardiol, D; de Ruiz Holgado, A P; de Mendoza, D

1990-01-01

The citrate plasmid (Cit+ plasmid) from Lactococcus lactis subsp. lactis biovar diacetylactis was cloned into the EcoRI site of plasmid pUC18. This recombinant plasmid enabled Escherichia coli K-12 to transport and utilize citrate as a source of energy, indicating expression of the citrate permease from L. lactis biovar diacetylactis. The citrate permease was under the control of the lac promoter of pUC18. Genetic expression of the Cit+ plasmid in maxicells revealed that the plasmid encoded two polypeptides of 47 and 32 kilodaltons, determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Images PMID:2117878

DNA fingerprinting of Brassica juncea cultivars using microsatellite probes.

PubMed

Bhatia, S; Das, S; Jain, A; Lakshmikumaran, M

1995-09-01

The genetic variability in the Brassica juncea cultivars was detected by employing in-gel hybridization of restricted DNA to simple repetitive sequences such as (GATA)4, (GACA)4 and (CAC)5. The most informative probe/enzyme combination was (GATA)4/EcoRI, yielding highly polymorphic fingerprint patterns for the B. juncea cultivars. This technique was found to be dependable for establishing the variety specific patterns for most of the cultivars studied, a prerequisite for germplasm preservation. The results of the present study were compared with those reported in our earlier study in which random amplification of polymorphic DNA (RAPD) was used for assessing the genetic variability in the B. juncea cultivars.

Characterization of epidemic Neisseria meningitidis serogroup C strains in several Brazilian states.

PubMed Central

Sacchi, C T; Tondella, M L; de Lemos, A P; Gorla, M C; Berto, D B; Kumiochi, N H; Melles, C E

1994-01-01

Epidemic strains of the Neisseria meningitidis C:2b:P1.3 electrophoretic type 11 complex were responsible for an outbreak in Curitiba, Parana State, Brazil, from 1990 to 1991. Strains of this complex were also isolated in other Brazilian states and were responsible for a meningococcal disease epidemic in São Paulo State in 1990. Serotyping both with monoclonal antibodies and by multilocus enzyme electrophoresis was useful for typing these epidemic strains related to the increased incidence of meningococcal disease. The genetic similarity of members of the electrophoretic type 11 complex was confirmed by the ribotyping method by using EcoRI or ClaI endonuclease restriction enzymes. Images PMID:7929775

Comparative ribotyping of Staphylococcus intermedius isolated from members of the Canoidea gives possible evidence for host-specificity and co-evolution of bacteria and hosts.

PubMed

Aarestrup, F M

2001-07-01

A total of 41 Staphylococcus intermedius isolates were isolated from skin of healthy members of six phylogenetic groups within the Canoidea (the dog family, skunk subfamily, weasel subfamily, racoon family, red panda and bear family) of different geographical origin and compared by EcoRI ribotyping and cluster analysis. The S. intermedius isolates from the different families and subfamilies clustered together in separate groups, almost completely following the phylogenetic relationship of the animal hosts. These ribotype data indicate host-specificity of different types of S. intermedius and suggest co-evolution between the animal hosts within the Canoidea and S. intermedius.

κ chain monoallelic demethylation and the establishment of allelic exclusion

PubMed Central

Mostoslavsky, Raul; Singh, Nandita; Kirillov, Andrei; Pelanda, Roberta; Cedar, Howard; Chess, Andrew; Bergman, Yehudit

1998-01-01

Allelic exclusion in κ light-chain synthesis is thought to result from a feedback mechanism by which the expression of a functional κ light chain on the surface of the B cell leads to an intracellular signal that down-regulates the V(D)J recombinase, thus precluding rearrangement of the other allele. Whereas such a feedback mechanism clearly plays a role in the maintenance of allelic exclusion, here we provide evidence suggesting that the initial establishment of allelic exclusion involves differential availability of the two κ alleles for rearrangement. Analysis of κ+ B-cell populations and of individual κ+ B cells that have rearranged only one allele demonstrates that in these cells, critical sites on the rearranged allele are unmethylated, whereas the nonrearranged allele remains methylated. This pattern is apparently generated by demethylation that is initiated at the small pre-B cell stage, on a single allele, in a process that occurs prior to rearrangement and requires the presence in cis of both the intronic and 3′ κ enhancers. Taken together with data demonstrating that undermethylation is required for rearrangement, these results indicate that demethylation may actually underly the process of allelic exclusion by directing the initial choice of a single κ allele for rearrangement. PMID:9637682

Comparison of genomes of malignant catarrhal fever-associated herpesviruses by restriction endonuclease analysis.

PubMed

Shih, L M; Zee, Y C; Castro, A E

1989-01-01

The restriction endonuclease DNA cleavage patterns of eight isolates of malignant catarrhal fever-associated herpesviruses were examined using the restriction endonucleases HindIII and EcoRI. The eight viruses could be assigned to two distinct groups. Virus isolates from a blue wildebeest, a sika deer and an ibex had restriction endonuclease DNA cleavage patterns that were in general similar to each other. The restriction pattern of these three viruses was distinct from the other five. Of these five, four were isolated from a greater kudu, a white tailed wildebeest, a white bearded wildebeest, and a cape hartebeest. The fifth isolate C500, was isolated from a domestic cow with malignant catarrhal fever. These five viruses had similar DNA cleavage patterns.

rRNA gene restriction patterns as an epidemiological marker in nosocomial outbreaks of Staphylococcus aureus infections.

PubMed

Meugnier, H; Fernandez, M P; Bes, M; Brun, Y; Bornstein, N; Freney, J; Fleurette, J

1993-01-01

rRNA gene restriction patterns (ribotyping) were compared with phage typing, serotyping, enterotoxins and exfoliatin production in the analysis of 26 Staphylococcus aureus strains isolated from two different nosocomial outbreaks. Total DNA was cleaved by EcoRI restriction endonuclease. After agarose gel electrophoresis and Southern transfer, the hybridization of the membranes was done with radiolabelled 16S rRNA gene from Bacillus subtilis inserted into a plasmid vector. Six to 13 fragments were visualized. A core of common fragments was discerned for all strains tested. A full correlation between ribotyping and conventional markers was observed in only one of the outbreaks studied. In both outbreaks, ribotyping proved helpful in characterizing otherwise untypable strains.

Validation of use of whole-cell repetitive extragenic palindromic sequence-based PCR (REP-PCR) for typing strains belonging to the Acinetobacter calcoaceticus-Acinetobacter baumannii complex and application of the method to the investigation of a hospital outbreak.

PubMed Central

Snelling, A M; Gerner-Smidt, P; Hawkey, P M; Heritage, J; Parnell, P; Porter, C; Bodenham, A R; Inglis, T

1996-01-01

Acinetobacter spp. are being reported with increasing frequency as causes of nosocomial infection. In order to identify reservoirs of infection as quickly as possible, a rapid typing method that can differentiate epidemic strains from environmental and nonepidemic strains is needed. In 1993, a cluster of Acinetobacter baumannii isolates from five patients in the adult intensive therapy unit of our tertiary-care teaching hospital led us to develop and optimize a rapid repetitive extragenic palindromic sequence-based PCR (REP-PCR) typing protocol for members of the Acinetobacter calcoaceticus-A. baumannii complex that uses boiled colonies and consensus primers aimed at repetitive extragenic palindromic sequences. Four of the five patient isolates gave the same REP-PCR typing pattern as isolates of A. baumannii obtained from the temperature probe of a Bennett humidifier; the fifth isolate had a unique profile. Disinfection of the probe with 70% ethanol, as recommended by the manufacturer, proved ineffective, as A. baumannii with the same REP-PCR pattern was isolated from it 10 days after cleaning, necessitating a change in our decontamination procedure. Results obtained with REP-PCR were subsequently confirmed by ribotyping. To evaluate the discriminatory power (D) of REP-PCR for typing members of the A. calcoaceticus-A. baumannii complex, compared with that of ribotyping, we have applied both methods to a collection of 85 strains that included representatives of six DNA groups within the complex. Ribotyping using EcoRI digests yielded 53 patterns (D = 0.98), whereas 68 different REP-PCR patterns were observed (D = 0.99). By computer-assisted analysis of gel images, 74 patterns were observed with REP-PCR (D = 1.0). Overall, REP-PCR typing proved to be slightly more discriminatory than ribotyping. Our results indicate that REP-PCR typing used boiled colonies is a simple, rapid, and effective means of typing members of the A. calcoaceticus-A. baumannii complex. PMID:8727902

The construction, identification and partial characterization of plasmids containing guinea-pig milk protein complementary DNA sequences.

PubMed Central

Craig, R K; Hall, L; Parker, D; Campbell, P N

1981-01-01

A complementary DNA (cDNA) plasmid library has been constructed in the plasmid pAT153, using poly(A)-containing RNA isolated from the lactating guinea-pig mammary gland as the starting material. Double stranded cDNA was inserted into the EcoRI site of the plasmid using poly(dA . dT) tails, then transformed into Escherichia coli HB101. From the resulting colonies we have selected and partially characterized plasmids containing cDNA copies of the mRNAs for casein A, casein B, casein C and alpha-lactalbumin. However, the proportion containing casein C cDNA was exceptionally low, and these contained at best 60% of the mRNA sequence. Images Fig. 2. Fig. 3. Fig. 4. Fig. 5. PMID:7306038

Molecular typing and exopolysaccharide biosynthesis of Burkholderia cepacia isolates from a Portuguese cystic fibrosis center.

PubMed

Richau, J A; Leitão, J H; Correia, M; Lito, L; Salgado, M J; Barreto, C; Cescutti, P; Sá-Correia, I

2000-04-01

This work describes the first epidemiological survey of Burkholderia cepacia involved in pulmonary infections among the Portuguese population with cystic fibrosis (CF) who attended the major CF treatment Center in Lisbon at Sta. Maria Hospital from 1995 to the end of 1997. The characterization of the genomic relatedness of the isolates was based on the analysis of their ribopatterns (with EcoRI) followed by construction of a ribotype-based phylogenetic tree. This study was complemented with macrorestriction fragment analysis by pulsed-field gel electrophoresis. After optimization of the solid growth medium, we found that exopolysaccharide (EPS) production by B. cepacia CF isolates is not as rare a phenomenon as was thought before; indeed, 70% of the isolates examined were EPS producers.

Molecular Typing and Exopolysaccharide Biosynthesis of Burkholderia cepacia Isolates from a Portuguese Cystic Fibrosis Center

PubMed Central

Richau, João A.; Leitão, Jorge H.; Correia, Manuela; Lito, Luís; Salgado, Maria José; Barreto, Celeste; Cescutti, Paola; Sá-Correia, Isabel

2000-01-01

This work describes the first epidemiological survey of Burkholderia cepacia involved in pulmonary infections among the Portuguese population with cystic fibrosis (CF) who attended the major CF treatment Center in Lisbon at Sta. Maria Hospital from 1995 to the end of 1997. The characterization of the genomic relatedness of the isolates was based on the analysis of their ribopatterns (with EcoRI) followed by construction of a ribotype-based phylogenetic tree. This study was complemented with macrorestriction fragment analysis by pulsed-field gel electrophoresis. After optimization of the solid growth medium, we found that exopolysaccharide (EPS) production by B. cepacia CF isolates is not as rare a phenomenon as was thought before; indeed, 70% of the isolates examined were EPS producers. PMID:10747161

Light-Regulated Transcription of Genes Encoding Peridinin Chlorophyll a Proteins and the Major Intrinsic Light-Harvesting Complex Proteins in the Dinoflagellate Amphidinium carterae Hulburt (Dinophycae)1

PubMed Central

ten Lohuis, Michael R.; Miller, David J.

1998-01-01

In the dinoflagellate Amphidinium carterae, photoadaptation involves changes in the transcription of genes encoding both of the major classes of light-harvesting proteins, the peridinin chlorophyll a proteins (PCPs) and the major a/c-containing intrinsic light-harvesting proteins (LHCs). PCP and LHC transcript levels were increased up to 86- and 6-fold higher, respectively, under low-light conditions relative to cells grown at high illumination. These increases in transcript abundance were accompanied by decreases in the extent of methylation of CpG and CpNpG motifs within or near PCP- and LHC-coding regions. Cytosine methylation levels in A. carterae are therefore nonstatic and may vary with environmental conditions in a manner suggestive of involvement in the regulation of gene expression. However, chemically induced undermethylation was insufficient in activating transcription, because treatment with two methylation inhibitors had no effect on PCP mRNA or protein levels. Regulation of gene activity through changes in DNA methylation has traditionally been assumed to be restricted to higher eukaryotes (deuterostomes and green plants); however, the atypically large genomes of dinoflagellates may have generated the requirement for systems of this type in a relatively “primitive” organism. Dinoflagellates may therefore provide a unique perspective on the evolution of eukaryotic DNA-methylation systems. PMID:9576788

Transcriptional activity across the Epstein-Barr virus genome in Raji cells during latency and after induction of an abortive lytic cycle.

PubMed

Kirchner, E A; Bornkamm, G W; Polack, A

1991-10-01

We have studied the relative rate of transcription across the Epstein-Barr virus genome in the Burkitt's lymphoma cell line Raji by nuclear run-on analysis during latency and after induction of an abortive lytic cycle with 12-0-tetradecanoylphorbol 13-acetate (TPA) and 5-iodo-2'-deoxyuridine (IUdR). During latency the entire, or almost the entire, viral genome was found to be transcriptionally active to a low or intermediate extent, with some variation in activity along the genome. The fragment with the highest transcriptional activity was EcoRI J, which contains the genes encoding the small nuclear RNAs EBER1 and -2, transcribed predominantly by RNA polymerase III. An intermediate level of transcription was observed between positions 10 and 138 (kb), with areas of slightly higher activity on the large internal repeats and the left duplicated region (DL). The remaining part of the viral genome, between position 138 and the termini, and the termini and position 10 (kb) (with the exception of the EcoRI J fragment), showed very little transcriptional activity, except for the intermediately active regions carrying the righthand oriLyt (DR) and the terminal repeats. Upon induction of the viral genome with TPA and IUdR, the viral genome was transcriptionally active at a rate at least tenfold that seen during latency. Polymerases were not equally distributed along the genome after induction; the highest density was found in regions 48 to 58 kb, 82 to 84 kb, 102 to 104 kb, 118 to 122 kb and 142 to 145 kb of the viral genome. High transcriptional activity correlated with distinct transcription units in some cases, i.e. BamHI H1LF1 (DL), BamHI MLF1, BamHI ZLF1/BamHI RLF1 and BamHI X (thymidine kinase), but not in others (BamHI H2). Besides initiation of transcription, other regulatory processes such as stabilization and processing of primary transcripts may also contribute to regulation of virus gene expression. Addition of cycloheximide completely abolished the transcriptional activation of the genome mediated by TPA and IUdR.

Highlights of the DNA cutters: a short history of the restriction enzymes

PubMed Central

Loenen, Wil A. M.; Dryden, David T. F.; Raleigh, Elisabeth A.; Wilson, Geoffrey G.; Murray, Noreen E.

2014-01-01

In the early 1950’s, ‘host-controlled variation in bacterial viruses’ was reported as a non-hereditary phenomenon: one cycle of viral growth on certain bacterial hosts affected the ability of progeny virus to grow on other hosts by either restricting or enlarging their host range. Unlike mutation, this change was reversible, and one cycle of growth in the previous host returned the virus to its original form. These simple observations heralded the discovery of the endonuclease and methyltransferase activities of what are now termed Type I, II, III and IV DNA restriction-modification systems. The Type II restriction enzymes (e.g. EcoRI) gave rise to recombinant DNA technology that has transformed molecular biology and medicine. This review traces the discovery of restriction enzymes and their continuing impact on molecular biology and medicine. PMID:24141096

Highlights of the DNA cutters: a short history of the restriction enzymes.

PubMed

Loenen, Wil A M; Dryden, David T F; Raleigh, Elisabeth A; Wilson, Geoffrey G; Murray, Noreen E

2014-01-01

In the early 1950's, 'host-controlled variation in bacterial viruses' was reported as a non-hereditary phenomenon: one cycle of viral growth on certain bacterial hosts affected the ability of progeny virus to grow on other hosts by either restricting or enlarging their host range. Unlike mutation, this change was reversible, and one cycle of growth in the previous host returned the virus to its original form. These simple observations heralded the discovery of the endonuclease and methyltransferase activities of what are now termed Type I, II, III and IV DNA restriction-modification systems. The Type II restriction enzymes (e.g. EcoRI) gave rise to recombinant DNA technology that has transformed molecular biology and medicine. This review traces the discovery of restriction enzymes and their continuing impact on molecular biology and medicine.

Restriction fragment length polymorphism and allozyme linkage map of Cuphea lanceolata.

PubMed

Webb, D M; Knapp, S J; Tagliani, L A

1992-02-01

Cuphea lanceolata Ait. has had a significant role in the domestication of Cuphea and is a useful experimental organism for investigating how medium-chain lipids are synthesized in developing seeds. To expand the genetics of this species, a linkage map of the C. lanceolata genome was constructed using five allozyme and 32 restriction-fragment-length-polymorphism (RFLP) marker loci. These loci were assigned to six linkage groups that correspond to the six chromosomes of this species. Map length is 288 cM. Levels of polymorphism were estimated for three inbred lines of C. lanceolata and an inbred line of C. viscosissima using 84 random genomic clones and two restriction enzymes, EcoRI and HindIII. Of the probes 29% detected RFLPs between C. lanceolata and C. viscosissima lines. Crosses between these species can be exploited to expand the map.

DNA fusion product of phage P2 with plasmid pBR322 - A new phasmid

NASA Technical Reports Server (NTRS)

Nicoletti, M.; Bertani, G.

1983-01-01

The chromosome of the temperate bacteriophage P2 and that of the plasmid pBR322 have been joined in vitro after treatment with restriction endonuclease EcoRI. The fusion product - a phasmid - can behave as a plasmid, as a phage and as a prophage. It can replicate its DNA under the control of either the specific replication mechanism of the parent phage in a polA mutant or that of the parent plasmid in a rep mutant. Several interesting interactions between the two replication modes are indicated. In particular, phage particles may be produced even when the phage mode of DNA replication is blocked, and this throws new light on the involvement of the early gene A in the regulation of late gene expression in phage P2.

Restriction fragment length polymorphism of the human c-fms gene.

PubMed Central

Xu, D Q; Guilhot, S; Galibert, F

1985-01-01

By using blot hybridization with a v-fms probe, a polymorphism for EcoRI, HindIII, and BamHI restriction endonuclease sites associated with the human c-fms locus was observed in a random adult population. This restriction fragment length polymorphism can be explained on the basis of the existence of two alleles, a and b, and is due to a short (congruent to 500 base pairs) deletion characteristic of allele a. The distribution in the analyzed population (48 unrelated individuals) is 23% heterozygotes ab, 75% homozygotes bb, and 2% homozygotes aa. Though the inheritance of this polymorphism follows a Mendelian pattern, the children from couples ab X bb are of the following genotype: 74% ab and 26% bb. These deviations from the expected frequencies of 50% suggest a selective pressure in favor of heterozygotes. Images PMID:2986142

Identification of a 'Candidatus Phytoplasma hispanicum'-related strain, associated with yellows-type diseases, in smoke-tree sharpshooter (Homalodisca liturata Ball).

PubMed

Servín-Villegas, Rosalía; Caamal-Chan, Maria Goretty; Chavez-Medina, Alicia; Loera-Muro, Abraham; Barraza, Aarón; Medina-Hernández, Diana; Holguín-Peña, Ramón Jaime

2018-04-11

The 16SrXIII group from phytoplasma bacteria were identified in salivary glands from Homalodisca liturata, which were collected in El Comitán on the Baja California peninsula in Mexico. We were able to positively identify 15 16S rRNA gene sequences with the corresponding signature sequence of 'CandidatusPhytoplasma' (CAAGAYBATKATGTKTAGCYGGDCT) and in silico restriction fragment length polymorphism (RFLP) profiles (F value estimations) coupled with a phylogenetic analysis to confirm their relatedness to 'CandidatusPhytoplasma hispanicum', which in turn belongs to the 16SrXIII group. A restriction analysis was carried out with AluI and EcoRI to confirm that the five sequences belongs to subgroup D. The rest of the sequences did not exhibit any known RFLP profile related to a subgroup reported in the 16SrXIII group.

Transformation of primary human embryonic kidney cells to anchorage independence by a combination of BK virus DNA and the Harvey-ras oncogene

DOE Office of Scientific and Technical Information (OSTI.GOV)

Pater, A.; Pater, M.M.

Primary human embryonic kidney (HEK) cells were transformed by a focus assay with BK virus (BKV) DNA molecularly cloned at its unique EcoRI site. Both viral DNA sequences and viral tumor antigens were present and expressed in all the foci that the authors examined. However, cells isolated from foci were incapable of growth in soft agar. They then examined the transformation of HEK cells after their transfection with a combination of BKV DNA and either the normal or the activated form of the human Ha-ras oncogene (EJ c-Ha-ras-1). Only the cells transfected with a combination of BKV DNA and themore » activated form of Ha-ras DNAs were present in the transformed colonies. BKV tumor antigens and the Ha-ras p21 protein were also expressed.« less

Physical and genetic mapping of the dipeptidase gene DPEP1 to 16q24. 3

DOE Office of Scientific and Technical Information (OSTI.GOV)

Austruy, E.; Jeanpierre, C.; Junien, C.

1993-03-01

The authors report the subregional physical and genetic mapping on chromosome 16q of a cDNA clone selected as a potential tumor/growth suppressor sequence. By DNA sequencing and RNA expression pattern, this clone was identified as part of the renal dipeptidase gene (DPEP1). Using somatic cell hybrids carrying either different human chromosomes or chromosome 16 segments, they confirm and refine the physical mapping of DPEP1 to the chromosome 16 subregion q24.3. Two RFLPs, a biallelic polymorphism detected by TaqI and a VNTR detected by BamHI, EcoRI, and BglII, are described. Using the VNTR polymorphism, DPEP1 was shown to be linked tomore » D16S7 with a maximum lod score of 5.8 at a recombination fraction of 0.03. 14 refs., 2 figs., 2 tabs.« less

Herpesvirus papio contains a plasmid origin of replication that acts in cis interspecies with an Epstein-Barr virus trans-acting function.

PubMed Central

Pesano, R L; Pagano, J S

1986-01-01

Herpesvirus papio (HVP) and Epstein-Barr virus (EBV) are closely related biologically and biochemically; lymphoblastoid cells infected with either virus contain episomal viral DNA. The putative origin of replication for EBV plasmids (oriP) has been assigned to a 1,790-base-pair fragment (cis) in the short unique region of the genome which requires a viral function supplied in trans from elsewhere in the genome (J. Yates, N. Warren, D. Reisman, and B. Sugden, Proc. Natl. Acad. Sci. USA 81:3806-3810, 1984). We report here the identification of the putative origin of replication (cis) in HVP; we assigned it to the HVP EcoRI K fragment. The results indicate that the HVP replication process requires both a cis and a trans-acting function, analogous to that found in EBV. Images PMID:3023667

Polymorphic human somatostatin gene is located on chromosome 3.

PubMed Central

Naylor, S L; Sakaguchi, A Y; Shen, L P; Bell, G I; Rutter, W J; Shows, T B

1983-01-01

Somatostatin is a 14-amino-acid neuropeptide and hormone that inhibits the secretion of several peptide hormones. The human gene for somatostatin SST has been cloned, and the sequence has been determined. This clone was used as a probe in chromosome mapping studies to detect the human somatostatin sequence in human-rodent hybrids. Southern blot analysis of 41 hybrids, including some containing translocations of human chromosomes, placed SST in the q21 leads to qter region of chromosome 3. Human DNAs from unrelated individuals were screened for restriction fragment polymorphisms detectable by the somatostatin gene probe. Two polymorphisms were found: (i) an EcoRI variant located at the 3' end of the gene, found in Caucasian, U.S. Black, and Asian populations with a frequency of approximately 0.10 and (ii) a BamHI variant in the intron, which occurs in Caucasians at a frequency of 0.13. Images PMID:6133281

The construction of a synthetic Escherichia coli trp promoter and its use in the expression of a synthetic interferon gene.

PubMed Central

Windass, J D; Newton, C R; De Maeyer-Guignard, J; Moore, V E; Markham, A F; Edge, M D

1982-01-01

An 82 base pair DNA fragment has been synthesised which contains the E. coli trp promoter and operator sequences and also encodes the first Shine Dalgarno sequence of the trp operon. This DNA fragment is flanked by EcoRI and ClaI/TaqI cohesive ends and is thus easy to clone, transfer between vector systems and couple to genes to drive their expression. It has been cloned into plasmid pAT153, producing a convenient trp promoter vector. We have also joined the fragment to a synthetic IFN-alpha 1 gene, using synthetic oligonucleotides to generate a completely natural, highly efficient bacterial translation initiation signal on the promoter proximal side of the IFN gene. Plasmids carrying this construction enable E. coli cells to express IFN-alpha 1 almost constitutively and with significantly higher efficiency than from a lacUV5 promoter based system. Images PMID:6184675

Evaluation of (GTG)5-PCR for rapid identification of Streptococcus mutans.

PubMed

Svec, Pavel; Nováková, Dana; Zácková, Lenka; Kukletová, Martina; Sedlácek, Ivo

2008-11-01

Repetitive sequence-based polymerase chain reaction (PCR) fingerprinting using the (GTG)(5) primer was applied for fast screening of bacterial strains isolated from dental plaque of early childhood caries (ECC)-affected children. A group of 29 Gram-positive bacteria was separated into a homogeneous cluster together with Streptococcus mutans reference strains and constituted an aberrant branch after the numerical analysis of (GTG)(5)-PCR fingerprints. Automated ribotyping with EcoRI restriction enzyme (RiboPrinter microbial characterization system) revealed high genetic heterogeneity among the tested group and proved to be a good tool for strain-typing purposes. Further characterization of the studied strains was achieved by extensive phenotyping and whole-cell protein fingerprinting and confirmed all the strains as S. mutans representatives. Obtained results showed rep-PCR fingerprinting with the (GTG)(5) primer to be a fast and reliable method for identification of S. mutans.

Isolation and characterization of two overlapping cosmid clones from the 4q35 region, near the facioscapulohumeral muscular dystrophy locus

DOE Office of Scientific and Technical Information (OSTI.GOV)

Deidda, G.; Grisanti, P.; Vigneti, E.

1994-09-01

The gene for facioscapulohumeral muscular dystrophy (FSHD) has been localized by linkage analysis to the 4q35 region. The most telomeric p13E-11 prove has been shown to detect 4q35 DNA rearrangements in both sporadic and familial cases of the disease. With the aim of constructing a detailed physical map of the 4q35 region and searching for the mutant gene, we used p13E-11 probe to isolate cosmid clones from a human genomic library in a pCos-EMBL 2 vector. Two positive clones were isolated, clones 3 and 5, which partially overlap and carry human genomic inserts of 42 and 45 kb, respectively. Themore » cosmids share a common region containing the p13E-11 region and a stretch of KpnI units consisting of 3.2 kb tandemly repeated sequences (about 10). The restriction maps were constructed using the following enzymes: Bam HI, BgIII, Eco RI, EcoRV, KpnI and Sfi I. Clone 3 extends 4 kb upstream of C5 and stops within the Kpn repeats. Clone 5 extends 4 kb downstream from the Kpn repeats and it presents an additional EcoRI site. Clone 5 contains a stretch of Kpn sequences of nearly 32 kb, corresponding to 10 Kpn repeats; clone 3 contains a stretch of 29 kb corresponding to 9 Kpn repeats, as determined by PFGE analysis of partial digestion of the clones. Clone 5 seems to contain the entire Eco RI region prone to rearrangements in FSHD patients. From clone 5 several subclones were obtained, from the Kpn region and from the region spanning from the last Kpn repeat to the cloning site. No single copy sequences were detected. Subclones from the 3{prime} end region contain beta-satellite or Sau3A-like sequences. In situ hybridization with the whole C5 cosmid shows hybridization signals at the tip of chromosome 4 (4q35) and chromosome 10 (10q26), in the pericentromeric region of chromosome 1 (1q12) and in the p12 region of the acrocentric chromosomes (chr. 21, 22, 13, 14, 15).« less

Resistance of Spiroplasma citri Lines to the Virus SVTS2 Is Associated with Integration of Viral DNA Sequences into Host Chromosomal and Extrachromosomal DNA.

PubMed

Sha, Y; Melcher, U; Davis, R E; Fletcher, J

1995-11-01

Spiroplasmavirus SVTS2, isolated from Spiroplasma melliferum TS2, produces plaques when inoculated onto lawns of Spiroplasma citri M200H, a derivative of the type strain Maroc R8A2. S. citri strains MR2 and MR3, originally selected as colonies growing within plaques on a lawn of M200H inoculated with SVTS2, were resistant to SVTS2. Genomic DNA fingerprints and electrophoretic protein profiles of M200H, MR2, and MR3 were similar, but three proteins present in M200H were missing or significantly reduced in both resistant lines. None of these three polypeptides reacted with antiserum against S. citri membrane proteins, indicating that they probably are not surface-located virus receptors. Electroporation with SVTS2 DNA produced 1.5 x 10(sup5) transfectants per (mu)g of DNA in M200H but none in MR2 or MR3, suggesting that resistance may result from inhibition of viral replication. The digestion patterns of the extrachromosomal double-stranded (ds) DNA of these lines were similar. Three TaqI fragments of MR2 extrachromosomal DNA that were not present in M200H extrachromosomal DNA hybridized strongly to an SVTS2 probe, and two of these fragments plus an additional one hybridized with the MR3 extrachromosomal DNA, indicating that a fragment of SVTS2 DNA was present in the extrachromosomal ds DNA of MR2 and MR3 but not of M200H. When the restricted genomes of all three lines were probed with SVTS2 DNA, strong hybridization to two EcoRI fragments of chromosomal MR2 and MR3 DNA but not M200H DNA indicated that SVTS2 DNA had integrated into the genomes of MR2 and MR3 but not of M200H. When MR3 extrachromosomal ds DNA containing a 2.1-kb SVTS2 DNA fragment was transfected into M200H, the transformed spiroplasmas were resistant to SVTS2. These results suggest that SVTS2 DNA fragments, possibly integrated into the chromosomal or extrachromosomal DNA of a previously susceptible spiroplasma, may function as viral incompatibility elements, providing resistance to superinfection by SVTS2.

Identification of the gene encoding a 38-kilodalton immunogenic and protective antigen of Streptococcus suis.

PubMed

Okwumabua, Ogi; Chinnapapakkagari, Sharmila

2005-04-01

In our continued effort to search for a Streptococcus suis protein(s) that can serve as a vaccine candidate or a diagnostic reagent, we constructed and screened a gene library with a polyclonal antibody raised against the whole-cell protein of S. suis type 2. A clone that reacted with the antibody was identified and characterized. Analysis revealed that the gene encoding the protein is localized within a 2.0-kbp EcoRI DNA fragment. The nucleotide sequence contained an open reading frame that encoded a polypeptide of 445 amino acid residues with a calculated molecular mass of 46.4 kDa. By in vitro protein synthesis and Western blot experiments, the protein exhibited an electrophoretic mobility of approximately 38 kDa. At the amino acid level the deduced primary sequence shared homology with sequences of unknown function from Streptococcus pneumoniae (89%), Streptococcus mutans (86%), Lactococcus lactis (80%), Listeria monocytogenes (74%), and Clostridium perfringens (64%). Except for strains of serotypes 20, 26, 32, and 33, Southern hybridization analysis revealed the presence of the gene in strains of other S. suis serotypes and demonstrated restriction fragment length differences caused by a point mutation in the EcoRI recognition sequence. We confirmed expression of the 38-kDa protein in the hybridization-positive isolates using specific antiserum against the purified protein. The recombinant protein was reactive with serum from pigs experimentally infected with virulent strains of S. suis type 2, suggesting that the protein is immunogenic and may serve as an antigen of diagnostic importance for the detection of most S. suis infections. Pigs immunized with the recombinant 38-kDa protein mounted antibody responses to the protein and were completely protected against challenge with a strain of a homologous serotype, the wild-type virulent strain of S. suis type 2, suggesting that it may be a good candidate for the development of a vaccine that can be used as protection against S. suis infection. Analysis of the cellular fractions of the bacterium by Western blotting revealed that the protein was present in the surface and cell wall extracts. The functional role of the protein with respect to pathogenesis and whether antibodies against the antigen confer protective immunity against diseases caused by strains of other pathogenic S. suis capsular types remains to be determined.

Extrachromosomal DNA length and antibiograms of Staphylococcus aureus and Pseudomonas aeruginosa isolated from tears of HIV/AIDS patients after curing with sodium dodecyl sulphate.

PubMed

Ajayi, B O; Otajevwo, F D

2012-01-01

Staphylococcus aureus and Pseudomonas aeruginosa strains were isolated from eye swab samples randomly obtained from 100 seropositive HIV/AIDS patients who reported to various anti-retroviral treatment clinics at the University of Benin Teaching Hospital and Central Hospital both based in Benin City, Nigeria. Invitro antibiotic sensitivity patterns of strains before curing were determined by the Kirby-Bauer disc diffusion technique. Resistance plasmid DNA of multidrug resistant strains was cured with 0.1% sodium dodecyl sulphate and cured strains were again subjected to in vitro antibiotic sensitivity testing. EcoRI and Hind III restriction endonuclease enzymes were used to make cuts on extracted plasmid DNA whose length sizes were then determined. A total of 36 (36.0%) strains made up of 27 (75.0%) Staphylococcus. aureus and 9 (25.0%) Pseudomonas aeruginosa were isolated of which 7 (19.4%) strains showed multidrug resistance to ciprofloxacin, pefloxacin, ofloxacine, gentamycin, tetracycline, ampicillin, chloramphenicol, nitrofurantoin and erythromycin. All seven multidrug resistant strains before curing, recorded 85.7%, 42.9%, 14.3% and 14.3% sensitivity in that decreasing order to ciprofloxacin, pefloxacin, ofloxacin and gentamycin respectively. There was 0.0% sensitivity each to tetracycline and ampicillin. After curing, there was enhanced sensitivity of 100.0%, 85.7%, 28.6% and 71.4% respectively. There was also 28.6% and 57.1% improved sensitivity to tetracycline and ampicillin after curing. Before curing, there was 76.2% average resistance to all used antibiotics and this reduced to 47.6% after curing Staph. aureus plasmid DNA. In the case of Pseudomonas aeruginosa, there was an average resistance of 76.3% before curing which fell to 42.5% after curing. EcoRI restriction enzyme gave the plasmid DNA length of Staphylococcus aureus strain 04 as 4.0Kb and this size depended upon the distance between recognition sites. Isolation of 36 (36.0%) strains of both isolates from 100 eye swabs shows the danger these organisms portend to all categories of opticians. The cheapness and high sensitivity of gentamycin justifies its use as eye drops for treatment of some eye infections. Curing of plasmid DNA is an indication that if SDS is administered to the organisms in sublethal doses, it can lead to the elimination of plasmid DNA without adverse effect on the genomic DNA of the bacterial strains.

Extrachromosomal DNA Length and Antibiograms of Staphylococcus aureus and Pseudomonas aeruginosa Isolated from Tears of HIV/AIDS Patients after Curing with Sodium Dodecyl Sulphate

PubMed Central

B. O., Ajayi; F. D., Otajevwo

2012-01-01

Staphylococcus aureus and Pseudomonas aeruginosa strains were isolated from eye swab samples randomly obtained from 100 seropositive HIV/AIDS patients who reported to various anti-retroviral treatment clinics at the University of Benin Teaching Hospital and Central Hospital both based in Benin City, Nigeria. Invitro antibiotic sensitivity patterns of strains before curing were determined by the Kirby-Bauer disc diffusion technique. Resistance plasmid DNA of multidrug resistant strains was cured with 0.1% sodium dodecyl sulphate and cured strains were again subjected to invitro antibiotic sensitivity testing. EcoRI and Hind III restriction endonuclease enzymes were used to make cuts on extracted plasmid DNA whose length sizes were then determined. A total of 36 (36.0%) strains made up of 27 (75.0%) Staphylococcus. aureus and 9 (25.0%) Pseudomonas aeruginosa were isolated of which 7 (19.4%) strains showed multidrug resistance to ciprofloxacin, pefloxacin, ofloxacine, gentamycin, tetracycline, ampicillin, chloramphenicol, nitrofurantoin and erythromycin. All seven multidrug resistant strains before curing, recorded 85.7%, 42.9%, 14.3% and 14.3% sensitivity in that decreasing order to ciprofloxacin, pefloxacin, ofloxacin and gentamycin respectively. There was 0.0% sensitivity each to tetracycline and ampicillin. After curing, there was enhanced sensitivity of 100.0%, 85.7%, 28.6% and 71.4% respectively. There was also 28.6% and 57.1% improved sensitivity to tetracycline and ampicillin after curing. Before curing, there was 76.2% average resistance to all used antibiotics and this reduced to 47.6% after curing Staph. aureus plasmid DNA. In the case of Pseudomonas aeruginosa, there was an average resistance of 76.3% before curing which fell to 42.5% after curing. EcoRI restriction enzyme gave the plasmid DNA length of Staphylococcus aureus strain 04 as 4.0Kb and this size depended upon the distance between recognition sites. Isolation of 36 (36.0%) strains of both isolates from 100 eye swabs shows the danger these organisms portend to all categories of opticians. The cheapness and high sensitivity of gentamycin justifies its use as eye drops for treatment of some eye infections. Curing of plasmid DNA is an indication that if SDS is administered to the organisms in sublethal doses, it can lead to the elimination of plasmid DNA without adverse effect on the genomic DNA of the bacterial strains. PMID:22980121

Effect of Acylglycerol Composition and Fatty Acyl Chain Length on Lipid Digestion in pH-Stat Digestion Model and Simulated In Vitro Digestion Model.

PubMed

Qi, Jin F; Jia, Cai H; Shin, Jung A; Woo, Jeong M; Wang, Xiang Y; Park, Jong T; Hong, Soon T; Lee, K-T

2016-02-01

In this study, a pH-stat digestion model and a simulated in vitro digestion model were employed to evaluate the digestion degree of lipids depending on different acylglycerols and acyl chain length (that is, diacylglycerol [DAG] compared with soybean oil representing long-chain triacylglycerol compared with medium-chain triacylglycerol [MCT]). In the pH-stat digestion model, differences were observed among the digestion degrees of 3 oils using digestion rate (k), digestion half-time (t1/2 ), and digestion extent (Φmax). The results showed the digestion rate order was MCT > soybean oil > DAG. Accordingly, the order of digestion half-times was MCT < soybean oil < DAG. In simulated in vitro digestion model, digestion rates (k') and digestion half-times (t'1/2 ) were also obtained and the results showed a digestion rate order of MCT (k' = 0.068 min(-1) ) > soybean oil (k' = 0.037 min(-1) ) > DAG (k' = 0.024 min(-1) ). Consequently, the order of digestion half-times was MCT (t'1/2 = 10.20 min) < soybean oil (t'1/2 = 18.74 min) < DAG (t'1/2 = 29.08 min). The parameters obtained using the 2 models showed MCT was digested faster than soybean oil, and that soybean oil was digested faster than DAG. © 2015 Institute of Food Technologists®

Digestibility of gluten proteins is reduced by baking and enhanced by starch digestion

PubMed Central

Pan, Xiaoyan; Bellido, Vincent; Toole, Geraldine A.; Gates, Fred K.; Wickham, Martin S. J.; Shewry, Peter R.; Bakalis, Serafim; Padfield, Philip; Mills, E. N. Clare

2015-01-01

Scope Resistance of proteins to gastrointestinal digestion may play a role in determining immune‐mediated adverse reactions to foods. However, digestion studies have largely been restricted to purified proteins and the impact of food processing and food matrices on protein digestibility is poorly understood. Methods and results Digestibility of a total gliadin fraction (TGF), flour (cv Hereward), and bread was assessed using in vitro batch digestion with simulated oral, gastric, and duodenal phases. Protein digestion was monitored by SDS‐PAGE and immunoblotting using monoclonal antibodies specific for celiac‐toxic sequences (QQSF, QPFP) and starch digestion by measuring undigested starch. Whereas the TGF was rapidly digested during the gastric phase the gluten proteins in bread were virtually undigested and digested rapidly during the duodenal phase only if amylase was included. Duodenal starch digestion was also slower in the absence of duodenal proteases. Conclusion The baking process reduces the digestibility of wheat gluten proteins, including those containing sequences active in celiac disease. Starch digestion affects the extent of protein digestion, probably because of gluten‐starch complex formation during baking. Digestion studies using purified protein fractions alone are therefore not predictive of digestion in complex food matrices. PMID:26202208

[Anaerobic digestion of lignocellulosic biomass with animal digestion mechanisms].

PubMed

Wu, Hao; Zhang, Pan-Yue; Guo, Jian-Bin; Wu, Yong-Jie

2013-02-01

Lignocellulosic material is the most abundant renewable resource in the earth. Herbivores and wood-eating insects are highly effective in the digestion of plant cellulose, while anaerobic digestion process simulating animal alimentary tract still remains inefficient. The digestion mechanisms of herbivores and wood-eating insects and the development of anaerobic digestion processes of lignocellulose were reviewed for better understanding of animal digestion mechanisms and their application in design and operation of the anaerobic digestion reactor. Highly effective digestion of lignocellulosic materials in animal digestive system results from the synergistic effect of various digestive enzymes and a series of physical and biochemical reactions. Microbial fermentation system is strongly supported by powerful pretreatment, such as rumination of ruminants, cellulase catalysis and alkali treatment in digestive tract of wood-eating insects. Oxygen concentration gradient along the digestive tract may stimulate the hydrolytic activity of some microorganisms. In addition, the excellent arrangement of solid retention time, digesta flow and end product discharge enhance the animal digestion of wood cellulose. Although anaerobic digestion processes inoculated with rumen microorganisms based rumen digestion mechanisms were developed to treat lignocellulose, the fermentation was more greatly limited by the environmental conditions in the anaerobic digestion reactors than that in rumen or hindgut. Therefore, the anaerobic digestion processes simulating animal digestion mechanisms can effectively enhance the degradation of wood cellulose and other organic solid wastes.

Digestibility of gluten proteins is reduced by baking and enhanced by starch digestion.

PubMed

Smith, Frances; Pan, Xiaoyan; Bellido, Vincent; Toole, Geraldine A; Gates, Fred K; Wickham, Martin S J; Shewry, Peter R; Bakalis, Serafim; Padfield, Philip; Mills, E N Clare

2015-10-01

Resistance of proteins to gastrointestinal digestion may play a role in determining immune-mediated adverse reactions to foods. However, digestion studies have largely been restricted to purified proteins and the impact of food processing and food matrices on protein digestibility is poorly understood. Digestibility of a total gliadin fraction (TGF), flour (cv Hereward), and bread was assessed using in vitro batch digestion with simulated oral, gastric, and duodenal phases. Protein digestion was monitored by SDS-PAGE and immunoblotting using monoclonal antibodies specific for celiac-toxic sequences (QQSF, QPFP) and starch digestion by measuring undigested starch. Whereas the TGF was rapidly digested during the gastric phase the gluten proteins in bread were virtually undigested and digested rapidly during the duodenal phase only if amylase was included. Duodenal starch digestion was also slower in the absence of duodenal proteases. The baking process reduces the digestibility of wheat gluten proteins, including those containing sequences active in celiac disease. Starch digestion affects the extent of protein digestion, probably because of gluten-starch complex formation during baking. Digestion studies using purified protein fractions alone are therefore not predictive of digestion in complex food matrices. © 2015 The Authors. Molecular Nutrition & Food Research published by Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.

Hypervariable and highly divergent intron-exon organizations in the chordate Oikopleura dioica.

PubMed

Edvardsen, Rolf B; Lerat, Emmanuelle; Maeland, Anne Dorthea; Flåt, Mette; Tewari, Rita; Jensen, Marit F; Lehrach, Hans; Reinhardt, Richard; Seo, Hee-Chan; Chourrout, Daniel

2004-10-01

Oikopleura dioica is a pelagic tunicate with a very small genome and a very short life cycle. In order to investigate the intron-exon organizations in Oikopleura, we have isolated and characterized ribosomal protein EF-1alpha, Hox, and alpha-tubulin genes. Their intron positions have been compared with those of the same genes from various invertebrates and vertebrates, including four species with entirely sequenced genomes. Oikopleura genes, like Caenorhabditis genes, have introns at a large number of nonconserved positions, which must originate from late insertions or intron sliding of ancient insertions. Both species exhibit hypervariable intron-exon organization within their alpha-tubulin gene family. This is due to localization of most nonconserved intron positions in single members of this gene family. The hypervariability and divergence of intron positions in Oikopleura and Caenorhabditis may be related to the predominance of short introns, the processing of which is not very dependent upon the exonic environment compared to large introns. Also, both species have an undermethylated genome, and the control of methylation-induced point mutations imposes a control on exon size, at least in vertebrate genes. That introns placed at such variable positions in Oikopleura or C. elegans may serve a specific purpose is not easy to infer from our current knowledge and hypotheses on intron functions. We propose that new introns are retained in species with very short life cycles, because illegitimate exchanges including gene conversion are repressed. We also speculate that introns placed at gene-specific positions may contribute to suppressing these exchanges and thereby favor their own persistence.

Expression and localization of components of the histone deacetylases multiprotein repressory complexes in the mouse preimplantation embryo.

PubMed

Kantor, Boris; Makedonski, Kirill; Shemer, Ruth; Razin, Aharon

2003-12-01

DNA methylation had been implicated in the assembly of multiprotein repressory complexes that affect chromatin architecture thereby rendering genes inactive. Proteins containing methyl binding domains (MBDs) are major components of these complexes. MBD3 is a component of the HDAC associated chromatin remodeling complex Mi2/NuRD. The addition of MBD2 to the Mi2/NuRD complex creates MeCP1, a complex that is known to inactivate methylated promoters. The undermethylated state of the mouse preimplantation embryo prompted us to investigate the known repressory complexes at this developmental stage. We found individual components of Mi2/NuRD: MBD3, Mi2, HDAC1 and HDAC2 to be expressed from a very early stage of embryo development and to localize in close proximity with each other and with constitutive heterochromatin by the blastula stage. Expression of MBD2, a component of MeCP1, starts in the blastula stage. Then it is also found to be in proximity with heterochromatin (based on DAPI staining) and with MBD3, Mi2 and HDAC1. In contrast, expression of MeCP2, an MBD containing component of a third repressory complex (MeCP2/Sin3A), is not seen in the preimplantation embryo. Our results suggest that both Mi2/NuRD and MeCP1 complexes are already present at the very early stages of embryo development, while a MeCP2 complex is added to the arsenal of repressory complexes post-implantation at a stage when DNA methylation takes place.

Current perspective on assessing site of digestion in ruminants.

PubMed

Merchen, N R; Elizalde, J C; Drackley, J K

1997-08-01

The site of nutrient digestion in the gastrointestinal tract of ruminants affects nutrient availability and the nature of digestive end-products supplied to the host animal. Methods to study site of digestion in vivo provide a tool to obtain data that enhance the ability to interpret or predict animal responses to some feeding practices. Examples are discussed for which site of digestion data provided insights that might not have been evident from other approaches. The use of site of digestion techniques may provide interpretation regarding digestion of N by ruminants different from those derived from measurement of total tract N digestion. Site of digestion measurements have been particularly important in studying effects of heat processing of protein sources and in understanding the nature of digestion of N in high-quality, fresh forages. Moreover, site of digestion techniques have been instrumental in interpreting the influences of supplemental fat sources on ruminal digestion and ruminal biohydrogenation and small intestinal digestion of long-chain fatty acids.

The functional-morphological adaptive strategy of digestive organs of decapodiform cephalopods

PubMed Central

OMURA, Ayano; ENDO, Hideki

2015-01-01

The digestive organs in decapodiform cephalopod species morphologically vary by individual lifestyle. We examined the following six species of adult decapodiformes cephalopods representing different habitats: Todarodes pacificus, Loligo bleekeri, Loligo edulis, Watasenia scintillans (pelagic), Sepia lycidas and Euprymna morsei (benthic). L. bleekeri and L. edulis possess a bursiform cecal sac connected to the cecum. Pelagic species have a single digestive gland smaller than in benthic species. T. pacificus has an oval digestive gland larger than that of L. bleekeri and L. edulis, which possess withered-looking and smaller digestive glands. In contrast, the digestive glands in benthic species are paired. S. lycidas and E. morsei have well-developed and larger digestive glands than those of the pelagic species. Well-developed digestive duct appendages are found in benthic species. In qualification of the mass of digestive organs, pelagic species have smaller stomachs, digestive glands and digestive ducts’ appendages than benthic species. Because pelagic species need to swim, they may possess smaller stomachs and larger cecums for more rapid digestion. A smaller digestive gland may have the advantage of reducing the body weight in pelagic species for rapid swimming. In contrast, since benthic species require a longer time for digestion than pelagic species, they compact more food in their stomachs and absorb nutrients via more organs, such as the digestive grand and digestive duct appendages, in addition to cecum. PMID:26369293

The functional-morphological adaptive strategy of digestive organs of decapodiform cephalopods.

PubMed

Omura, Ayano; Endo, Hideki

2016-01-01

The digestive organs in decapodiform cephalopod species morphologically vary by individual lifestyle. We examined the following six species of adult decapodiformes cephalopods representing different habitats: Todarodes pacificus, Loligo bleekeri, Loligo edulis, Watasenia scintillans (pelagic), Sepia lycidas and Euprymna morsei (benthic). L. bleekeri and L. edulis possess a bursiform cecal sac connected to the cecum. Pelagic species have a single digestive gland smaller than in benthic species. T. pacificus has an oval digestive gland larger than that of L. bleekeri and L. edulis, which possess withered-looking and smaller digestive glands. In contrast, the digestive glands in benthic species are paired. S. lycidas and E. morsei have well-developed and larger digestive glands than those of the pelagic species. Well-developed digestive duct appendages are found in benthic species. In qualification of the mass of digestive organs, pelagic species have smaller stomachs, digestive glands and digestive ducts' appendages than benthic species. Because pelagic species need to swim, they may possess smaller stomachs and larger cecums for more rapid digestion. A smaller digestive gland may have the advantage of reducing the body weight in pelagic species for rapid swimming. In contrast, since benthic species require a longer time for digestion than pelagic species, they compact more food in their stomachs and absorb nutrients via more organs, such as the digestive grand and digestive duct appendages, in addition to cecum.

Plasmid origin of replication of herpesvirus papio: DNA sequence and enhancer function.

PubMed Central

Loeb, D D; Sung, N S; Pesano, R L; Sexton, C J; Hutchison, C; Pagano, J S

1990-01-01

Herpesvirus papio (HVP) is a lymphotropic virus of baboons which is related to Epstein-Barr virus (EBV) and produces latent infection. The nucleotide sequence of the 5,775-base-pair (bp) EcoRI K fragment of HVP, which has previously been shown to confer the ability to replicate autonomously, has been determined. Within this DNA fragment is a region which bears structural and sequence similarity to the ori-P region of EBV. The HVP ori-P region has a 10- by 26-bp tandem array which is related to the 20- by 30-bp tandem array from the EBV ori-P region. In HVP there is an intervening region of 764 bp followed by five partial copies of the 26-bp monomer. Both the EBV and HVP 3' regions have the potential to form dyad structures which, however, differ in arrangement. We also demonstrate that a transcriptional enhancer which requires transactivation by a virus-encoded factor is present in the HVP ori-P. Images PMID:2159548

Screening of cytoplasmic DNA diversity between and within Lupinus mutabilis Sweet and Lupinus albus sensu lato by restriction fragment length polymorphism (RFLP).

PubMed

Olczak, T; Rurek, M; Jańska, H; Augustyniak, H; Sawicka-Sienkiewicz, E J

2001-01-01

Seven populations and five mutant lines of the Andean lupin and four species from the section Albus were screened for their mitochondrial and chloroplast polymorphisms. For this purpose the RFLP method with EcoRI as a restriction enzyme was used. Lupinus luteus, Lupinus albus and Phaseolus vulgaris organellar clones as well as amplified fragments were used as probes. We found that mitochondrial probes were more suitable than chloroplast probes for identification of inter- and intra-specific variations within the examined material. Most mitochondrial probes differentiate the two species investigated. A high level of mitochondrial polymorphism was observed among the populations of L. mutabilis in contrast to monomorphism among the species in the section Albus. A limited polymorphism was detected between the mutant lines of L. mutabilis. We conclude from this study that the mitochondrial RFLP analysis is a valuable tool for identification of variability among Andean lupin populations.

Chromosomal arrangement of leghemoglobin genes in soybean.

PubMed Central

Lee, J S; Brown, G G; Verma, D P

1983-01-01

A cluster of four different leghemoglobin (Lb) genes was isolated from AluI-HaeIII and EcoRI genomic libraries of soybean in a set of overlapping clones which together include 45 kilobases (kb) of contiguous DNA. These four genes, including a pseudogene, are present in the same orientation and are arranged in the order: 5'-Lba-Lbc1-Lb psi-Lbc3-3'. The intergenic regions average 2.5 kb. In addition to this main Lb locus, there are other Lb genes which do not appear to be contiguous to this locus. A sequence probably common to the 3' region of Lb loci was found flanking the Lbc3 gene. The 3' flanking region of the main Lb locus also contains a sequence that appears to be expressed more abundantly in root tissue. Another sequence which is primarily expressed in root and leaf is found 5' to two Lb loci. Overall, the main leghemoglobin locus is similar in structure to the mammalian globin gene loci. Images PMID:6310504

Characterization of new plasmids from methylotrophic bacteria.

PubMed

Brenner, V; Holubová, I; Benada, O; Hubácek, J

1991-07-01

Several tens of methanol-utilizing bacterial strains isolated from soil were screened for the presence of plasmids. From the obligate methylotroph Methylomonas sp. strain R103a plasmid pIH36 (36 kb) was isolated and its restriction map was constructed. In pink-pigmented facultative methylotrophs (PPFM), belonging to the genus Methylobacterium four plasmids were detected: plasmids pIB200 (200 kb) and pIB14 (14 kb) in the strain R15d and plasmids pWU14 (14 kb) and pWU7 (7.8 kb) in the strain M17. Because of the small size and the presence of several unique REN sites (HindIII, EcoRI, NcoI), plasmid pWU7 was chosen for the construction of a vector for cloning in methylotrophs. Cointegrates pKWU7A and pKWU7B were formed between pWU7 and the E. coli plasmid pK19 Kmr, which were checked for conjugative transfer from E. coli into the methylotrophic host.

Yersinia pekkanenii sp. nov.

PubMed

Murros-Kontiainen, Anna; Johansson, Per; Niskanen, Taina; Fredriksson-Ahomaa, Maria; Korkeala, Hannu; Björkroth, Johanna

2011-10-01

The taxonomic position of three strains from water, soil and lettuce samples was studied by using a polyphasic taxonomic approach. The strains were reported to lack the virulence-encoding genes inv and virF in a previous study. Controversially, API 20 E and some other phenotypic tests suggested that the strains belong to Yersinia pseudotuberculosis, which prompted this polyphasic taxonomic study. In both the phylogenetic analyses of four housekeeping genes (glnA, gyrB, recA and HSP60) and numerical analyses of HindIII and EcoRI ribopatterns, the strains formed a separate group within the genus Yersinia. Analysis of the 16S rRNA gene sequences showed that the strains were related to Yersinia aldovae and Yersinia mollaretii, but DNA-DNA hybridization analysis differentiated them from these species. Based on the results of the phylogenetic and DNA-DNA hybridization analyses, a novel species, Yersinia pekkanenii sp. nov., is proposed. The type strain is ÅYV7.1KOH2(T) ( = DSM 22769(T)  = LMG 25369(T)).

Anaerobic Digestion and its Applications

EPA Science Inventory

Anaerobic digestion is a natural biological process. The initials "AD" may refer to the process of anaerobic digestion, or the built systems of anaerobic digesters. While there are many kinds of digesters, the biology is basically the same for all. Anaerobic digesters are built...

Life in the Clouds of Venus? An Experimental Synthetic Biology Approach

NASA Technical Reports Server (NTRS)

Rothschild, L. J.; Paulino-Lima, I. G.; Amatya, D.; Bajar, B.; Geilich, B.; Hu, J.; Jackson, C. J.

2015-01-01

The surface of Venus constitutes the most hellish and biologically inhospitable planetary surface in our solar system, boasting a pH of 0, blistering winds that can melt lead, and pressures of 60 atm. However, during the earlier years of the solar system, without the runaway greenhouse effect that has plagued the planet, Venus potentially housed oceans and perhaps even life. There is a possibility that microbes could have retreated into hospitable niches in the atmosphere, as suggested by Carl Sagan as early as 1967 [1]. For example, 50 km above the raging hell of the Venusian surface, exists a relatively temperate environment that might serve as reservoir for life. This astrobiology project seeks to explore life at the extremes and to theorize whether microbial communities could not only survive but also reproduce in the Venusian atmosphere. Specifically, we ask: are aerosols viable microbial environments? But before we can test for life in the clouds, we have to develop a proper reporter to visualize cell growth in situ. For this purpose, we aimed to develop cell-growth dependent reporters to serve as remote biosensors for cell growth. We developed two using the polA promoter, a DNA-replication dependent promoter, and nrd operon promoter, a cell-cycle dependent promoter. Using these cell-growth reporters, the next step is to aerosolize microbes expressing these reporters in a suspension chamber adapted from a Millikan Drop Apparatus to assay reproduction in an aerosolized environment. Better yet is to test the reproduction of microbes in a microgravity regime such as on ISS.Approach: We engineered two cell-cycle dependent genetic reporters. One was the polA promoter which codes for DNA Polymerase I, a gene active in DNA replication [2]. The other was the nrdP. The activation of ribonucleotide reductase reduces ribonucleotides into deoxyribonucleotides and is involved in the bacterial cell cycle [3]. This promoter began activation during the initiation of DNA replication and is cell-cycle dependent [4]. These promoters were fused to a GFP reporter, transformed into E. coli. The constructs were deposited in the iGEM registry as K847210: Escherichia coli DNA-replication dependent polA promoter K847211: Escherichia coli cell-division dependent nrd promoter Results: Our constructs displayed fluorescence when transformed into NEB-5alpha competent cells. While nrdP-E0840 displayed sufficient fluorescence as verified by fluorescent microscopy, the original polAP-E0840 construct (which uses mut3b GFP) exhibited low expression; while fluorescence was visible under the microscope, the signal was too weak for the camera to recognize. The polA promoter was therefore digest-ed with EcoRI and SpeI then ligated into plasmid pNCS containing a RBS, Clover, and a terminator. Clover is a highly engineered green fluorescent protein that exhibits extreme brightness [5] Fluoresence time course data demonsrated that the genes were induced in a cell cycle dependant manner [6]. Our assays via microscopy and the bulk assay shows that our promoters are functional as cell cycle reporters.Conclusions: The application of such tools are widespread and not limited to astrobiology; nrdP could be used to determine doubling times empirically and could possibly extrapolate DNA content from intensity of signals expressed by polAP. However, we are pri-marily interested in its use in astrobiology.

Improving biogas quality and methane yield via co-digestion of agricultural and urban biomass wastes.

PubMed

Poulsen, Tjalfe G; Adelard, Laetitia

2016-08-01

Impact of co-digestion versus mono-digestion on biogas and CH4 yield for a set of five biomass materials (vegetable food waste, cow dung, pig manure, grass clippings, and chicken manure) was investigated considering 95 different biomass mixes of the five materials under thermophilic conditions in bench-scale batch experiments over a period of 65days. Average biogas and CH4 yields were significantly higher during co-digestion than during mono-digestion of the same materials. This improvement was most significant for co-digestion experiments involving three biomass types, although it was independent of the specific biomasses being co-digested. Improvement in CH4 production was further more prominent early in the digestion process during co-digestion compared to mono-digestion. Co-digestion also appeared to increase the ultimate CH4/CO2 ratio of the gas produced compared to mono-digestion although this tendency was relatively weak and not statistically significant. Copyright © 2016 Elsevier Ltd. All rights reserved.

Anaerobic Digestion. Student Manual. Biological Treatment Process Control.

ERIC Educational Resources Information Center

Carnegie, John W., Ed.

This student manual contains the textual material for a four-lesson unit on anaerobic digestion control. Areas addressed include: (1) anaerobic sludge digestion (considering the nature of raw sludge, purposes of anaerobic digestion, the results of digestion, types of equipment, and other topics); (2) digester process control (considering feeding…

The function of digestive enzymes on Cu, Zn, and Pb release from soil in in vitro digestion tests.

PubMed

Li, Yi; Demisie, Walelign; Zhang, Ming-kui

2013-07-01

The bioaccessibility of soil heavy metals is the solubility of soil heavy metals in synthetic human digestive juice, which is usually determined using in vitro digestion test. To reveal the effects of digestive enzymes on soil heavy metals bioaccessibility, three representative in vitro digestion tests, Simple Bioaccessibility Extraction Test (SBET), Physiologically Based Extraction Test (PBET), and Simple Gastrointestinal Extraction Test (SGET), were chosen. The bioaccessibility of soil Cu, Zn, and Pb in each method were respectively evaluated with and without digestive enzymes, and the differences were compared. The results showed that the effects of digestive enzymes varied with different methods and elements. Because of digestive enzymes addition, the environmental change from acid gastric phase to neutral intestinal phase of PBET did not result in apparently decrease of the bioaccessibility of soil Cu. However, the solubility of soil Zn and Pb were pH-dependent. For SGET, when digestive enzymes were added, its results reflected more variations resulting from soil and element types. The impacts of digestive enzymes on heavy metal dissolution are mostly seen in the intestinal phase. Therefore, digestive enzyme addition is indispensable to the gastrointestinal digestion methods (PBET and SGET), while the pepsin addition is not important for the methods only comprised of gastric digestion (SBET).

Personalizing Protein Nourishment

PubMed Central

DALLAS, DAVID C.; SANCTUARY, MEGAN R.; QU, YUNYAO; KHAJAVI, SHABNAM HAGHIGHAT; VAN ZANDT, ALEXANDRIA E.; DYANDRA, MELISSA; FRESE, STEVEN A.; BARILE, DANIELA; GERMAN, J. BRUCE

2016-01-01

Proteins are not equally digestible—their proteolytic susceptibility varies by their source and processing method. Incomplete digestion increases colonic microbial protein fermentation (putrefaction), which produces toxic metabolites that can induce inflammation in vitro and have been associated with inflammation in vivo. Individual humans differ in protein digestive capacity based on phenotypes, particularly disease states. To avoid putrefaction-induced intestinal inflammation, protein sources and processing methods must be tailored to the consumer’s digestive capacity. This review explores how food processing techniques alter protein digestibility and examines how physiological conditions alter digestive capacity. Possible solutions to improving digestive function or matching low digestive capacity with more digestible protein sources are explored. Beyond the ileal digestibility measurements of protein digestibility, less invasive, quicker and cheaper techniques for monitoring the extent of protein digestion and fermentation are needed to personalize protein nourishment. Biomarkers of protein digestive capacity and efficiency can be identified with the toolsets of peptidomics, metabolomics, microbial sequencing and multiplexed protein analysis of fecal and urine samples. By monitoring individual protein digestive function, the protein component of diets can be tailored via protein source and processing selection to match individual needs to minimize colonic putrefaction and, thus, optimize gut health. PMID:26713355

Digestive Diseases

MedlinePlus

... cells and provide energy. This process is called digestion. Your digestive system is a series of hollow organs joined ... are also involved. They produce juices to help digestion. There are many types of digestive disorders. The ...

The dynamic changes and interactional networks of prokaryotic community between co-digestion and mono-digestions of corn stalk and pig manure.

PubMed

Wang, Min; Zhang, Xueying; Zhou, Jun; Yuan, Yuexiang; Dai, Yumei; Li, Dong; Li, Zhidong; Liu, Xiaofeng; Yan, Zhiying

2017-02-01

Anaerobic co-digestion is considered to be an efficient way to improve the biogas production. The abundance, dynamic and interactional networks of prokaryotic community were investigated between co-digestion and mono-digestions of corn stalk and pig manure in mesophilic batch test. Co-digestion showed higher methane production, and contributed to suitable microenvironment as well as stable prokaryotic community structure. The highest methane production was achieved with the highest relative abundance of Methanosaeta. Prokaryotic community in mono-digestions might inhibited by FA or FVFA. The functional modules in co-digestion and mono-digestion of pig manure clustered together with bigger size and higher degree, and the connections of metabolic functions were better-organized, which means high-efficient utilization of substrate and prevention of the two digestion systems crash. The partial mantel tests showed the functional modules were significantly affected by environmental factors. These results further explained that why co-digestion was more efficient than mono-digestion owing to suitable microenvironment. Copyright © 2016 Elsevier Ltd. All rights reserved.

Effects of Aging on the Digestive System

MedlinePlus

... switch to the Professional version Home Digestive Disorders Biology of the Digestive System Effects of Aging on ... Version. DOCTORS: Click here for the Professional Version Biology of the Digestive System Overview of the Digestive ...

Enhanced methane yield by co-digestion of sewage sludge with micro-algae and catering waste leachate.

PubMed

2018-04-04

The co-digestion of different wastes is a promising concept to improve methane generation during anaerobic process. However, the anaerobic co-digestion of catering waste leachate with algal biomass and sewage sludge has not been studied to date. This work investigated the methane generation by the anaerobic co-digestion of different mixtures of catering waste leachate, micro-algal biomass, and sewage sludge. Co-digestion of waste mixture containing equal ratios of three substrates had 39.31% higher methane yield than anaerobic digestion of raw sludge. This was possibly due to a proliferation of methanogens during the co-digestion period induced by multi-phase digestion of different wastes with different degrees of digestibility. Therefore, co-digestion of catering waste leachate, micro-algal biomass, and sewage sludge appears to be an efficient technology for energy conversion from waste resources. The scientific application of this co-digestion technology with these three substrates may play a role in solving important environmental issues of waste management.

Anaerobic digestion potential of urban organic waste: a case study in Malmö.

PubMed

Davidsson, Asa; Jansen, Jes la Cour; Appelqvist, Björn; Gruvberger, Christopher; Hallmer, Martin

2007-04-01

A study of existing organic waste types in Malmö, Sweden was performed. The purpose was to gather information about organic waste types in the city to be able to estimate the potential for anaerobic treatment in existing digesters at the wastewater treatment plan (WWTP). The urban organic waste types that could have a significant potential for anaerobic digestion amount to about 50 000 tonnes year(-1) (sludge excluded). Some of the waste types were further evaluated by methane potential tests and continuous pilot-scale digestion. Single-substrate digestion and co-digestion of pre-treated, source-sorted organic fraction of municipal solid waste, wastewater sludge, sludge from grease traps and fruit and vegetable waste were carried out. The experiments showed that codigestion of grease sludge and WWTP sludge was a better way of making use of the methane potential in the grease trap sludge than single-substrate digestion. Another way of increasing the methane production in sludge digesters is to add source-sorted organic fraction of municipal solid waste (SSOFMSW). Adding SSOFMSW (20% of the total volatile solids) gave a 10-15% higher yield than could be expected by comparison with separate digestion of sludge respective SSOFMSW. Co-digestion of sludge and organic waste is beneficial not just for increasing gas production but also for stabilizing the digestion process. This was seen when co-digesting fruit and vegetable waste and sludge. When co-digested with sludge, this waste gave a better result than the separate digestion of fruit and vegetable waste. Considering single-substrate digestion, SSOFMSW is the only waste in the study which makes up a sufficient quantity to be suitable as the base substrate in a full-scale digester that is separated from the sludge digestion. The two types of SSOFMSW tested in the pilot-scale digestion were operated successfully at mesophilic temperature. By adding SSOFMSW, grease trap sludge and fruit and vegetables waste to sludge digesters at the wastewater treatment plant, the yearly energy production from methane could be expected to increase from 24 to 43 GWh.

Tests of spool models for DNA packaging in phage lambda.

PubMed

Widom, J; Baldwin, R L

1983-12-25

Experiments are reported which bear on two spool models proposed for packaging the DNA of phage lambda. Both spool models fill an assumed spherical cavity with DNA wrapped in cylindrical or quasi-cylindrical layers composed of adjacent circular turns. In the curved-spool model, a single continuous segment of DNA, about 20% of the DNA length and probably located near the left end of the DNA, is in contact with the coat protein of the phage capsid. In the straight spool model, there are several DNA segments in contact with the capsid; they are concentrated in one half (probably the left half) of lambda DNA. We have identified the loci on the DNA which are in contact with the capsid by chemical crosslinking, induced by ultraviolet-irradiation of phage containing 5-bromodeoxyuridine in place of thymine. In an electron microscope experiment, phage are first lysed with EDTA, and then spread in a cytochrome c film by the formamide method. The disrupted capsid, which has the appearance of a phage ghost, serves as a marker showing where the DNA is crosslinked to the coat. The left end of the DNA is not distinguished from the right end, and so the map of DNA-capsid contacts is folded over on itself. Contacts are found nearly randomly over the entire map. In a second experiment, DNA from lysed, crosslinked phage is cut either with EcoRI or HindIII restriction endonucleases and the cut restriction fragments are labeled at their ends with 32P. Density centrifugation in a CsCl gradient separates free DNA from restriction fragments crosslinked to protein. After digestion with proteinase k, the DNA fragments previously crosslinked to protein are identified by size after agarose gel electrophoresis. DNA fragments from all parts of the genome are found. These two experiments show that, if the DNA of each phage is packaged identically, then the curved-spool model is ruled out and the straight spool model is unlikely. Alternatively, the manner of packaging the DNA may vary from one phage to the next. These results agree with other recent experiments on lambda DNA packaging by Hall & Schellman (1982a,b), and by Haas et al. (1982). A different experiment is also reported. The psoralen derivative aminomethyltrioxalen (AMT) is allowed to intercalate into lambda phage and then the DNA strands are crosslinked by ultraviolet-irradiation after the rapid phase of AMT intercalation is complete. The DNA is subsequently denatured by glyoxal modification and spread for electron microscopy in a cytochrome c film by the formamide method.(ABSTRACT TRUNCATED AT 400 WORDS)

Interactions between seed traits and digestive processes determine the germinability of bird-dispersed seeds

PubMed Central

Soons, Merel B.

2018-01-01

Waterbirds disperse a wide range of plant seeds via their guts, promoting biotic connectivity between isolated habitat patches. However, the intensity of digestive forces encountered by seeds, and therefore their potential to survive digestive tract passage, varies within and between waterbird species. Here, we investigate under controlled conditions how the interaction between seed traits and digestive strategies affect the germinability of seeds following waterbird-mediated dispersal. We exposed seeds of 30 wetland plant species to the main digestive processes in the dabbling duck digestive system: mechanical, chemical and intestinal digestion. These were simulated by 1) a pressure test and scarification treatment, 2) incubation in simulated gastric juice, and 3) incubation in intestinal contents of culled mallards (Anas platyrhynchos). We evaluated their separate and combined effects on seed germination, and identified the role of seed size and seed coat traits in resisting the digestive forces. Seeds were generally resistant to separate digestive processes, but highly sensitive to a combination. Resistance to mechanical break-down was reduced by up to 80% by chemical pre-treatment, especially for seeds with permeable coats. Scarified seeds were 12–17% more vulnerable to chemical and intestinal digestive processes than undamaged seeds. Large seeds and seeds with thin, permeable coats were particularly sensitive to chemical and intestinal digestion. These results indicate that efficient digestion of seeds requires multiple digestive processes. The gizzard, responsible for mechanical digestion, plays a key role in seed survival. Omnivorous birds, which have relatively light gizzards compared to pure herbivores or granivores, are thus most likely to disperse seeds successfully. Regardless of digestive strategy, small seeds with tough seed coats are most resistant to digestion and may be adapted to endozoochorous dispersal by waterbirds. PMID:29614085

Interactions between seed traits and digestive processes determine the germinability of bird-dispersed seeds.

PubMed

Kleyheeg, Erik; Claessens, Mascha; Soons, Merel B

2018-01-01

Waterbirds disperse a wide range of plant seeds via their guts, promoting biotic connectivity between isolated habitat patches. However, the intensity of digestive forces encountered by seeds, and therefore their potential to survive digestive tract passage, varies within and between waterbird species. Here, we investigate under controlled conditions how the interaction between seed traits and digestive strategies affect the germinability of seeds following waterbird-mediated dispersal. We exposed seeds of 30 wetland plant species to the main digestive processes in the dabbling duck digestive system: mechanical, chemical and intestinal digestion. These were simulated by 1) a pressure test and scarification treatment, 2) incubation in simulated gastric juice, and 3) incubation in intestinal contents of culled mallards (Anas platyrhynchos). We evaluated their separate and combined effects on seed germination, and identified the role of seed size and seed coat traits in resisting the digestive forces. Seeds were generally resistant to separate digestive processes, but highly sensitive to a combination. Resistance to mechanical break-down was reduced by up to 80% by chemical pre-treatment, especially for seeds with permeable coats. Scarified seeds were 12-17% more vulnerable to chemical and intestinal digestive processes than undamaged seeds. Large seeds and seeds with thin, permeable coats were particularly sensitive to chemical and intestinal digestion. These results indicate that efficient digestion of seeds requires multiple digestive processes. The gizzard, responsible for mechanical digestion, plays a key role in seed survival. Omnivorous birds, which have relatively light gizzards compared to pure herbivores or granivores, are thus most likely to disperse seeds successfully. Regardless of digestive strategy, small seeds with tough seed coats are most resistant to digestion and may be adapted to endozoochorous dispersal by waterbirds.

Prior lactose glycation of caseinate via the Maillard reaction affects in vitro activities of the pepsin-trypsin digest toward intestinal epithelial cells.

PubMed

Wang, X P; Zhao, X H

2017-07-01

The well-known Maillard reaction in milk occurs between lactose and milk proteins during thermal treatment, and its effects on milk nutrition and safety have been well studied. A lactose-glycated caseinate was prepared via this reaction and digested using 2 digestive proteases, pepsin and trypsin. The glycated caseinate digest was assessed for its in vitro activities on rat intestinal epithelial cells in terms of growth proliferation, anti-apoptotic effect, and differentiation induction using caseinate digest as reference, to verify potential effects of the Maillard reaction on these activities of caseinate digest to the cells. Two digests had proliferative and anti-apoptotic effects, and reached the highest effects at 0.02 g/L of digest concentration with treatment time of 24 h. In comparison with caseinate digest, glycated caseinate digest always showed weaker proliferative (5.3-14.2%) and anti-apoptotic (5.9-39.0%) effects, and was more toxic to the cells at 0.5 g/L of digest concentration with treatment time of 48 h. However, glycated caseinate digest at 2 incubation times of 4 to 7 d showed differentiation induction higher than caseinate digest, as it could confer the cells with increased activities in lactase (16.3-26.6%), sucrase (22.4-31.2%), and alkaline phosphatase (17.4-24.8%). Transmission electron microscopy observation results also confirmed higher differentiation induction of glycated caseinate digest. Amino acid loss and lactose glycation partially contributed to these decreased and enhanced activities of glycated caseinate digest, respectively. The Maillard reaction of caseinate and lactose is thus shown in this study to have effects on the activities of caseinate digest to intestinal epithelial cells. Copyright © 2017 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Fungal fermentation on anaerobic digestate for lipid-based biofuel production.

PubMed

Zhong, Yuan; Liu, Zhiguo; Isaguirre, Christine; Liu, Yan; Liao, Wei

2016-01-01

Anaerobic digestate is the effluent from anaerobic digestion of organic wastes. It contains a significant amount of nutrients and lignocellulosic materials, even though anaerobic digestion consumed a large portion of organic matters in the wastes. Utilizing the nutrients and lignocellulosic materials in the digestate is critical to significantly improve efficiency of anaerobic digestion technology and generate value-added chemical and fuel products from the organic wastes. Therefore, this study focused on developing an integrated process that uses biogas energy to power fungal fermentation and converts remaining carbon sources, nutrients, and water in the digestate into biofuel precursor-lipid. The process contains two unit operations of anaerobic digestion and digestate utilization. The digestate utilization includes alkali treatment of the mixture feed of solid and liquid digestates, enzymatic hydrolysis for mono-sugar release, overliming detoxification, and fungal fermentation for lipid accumulation. The experimental results conclude that 5 h and 30 °C were the preferred conditions for the overliming detoxification regarding lipid accumulation of the following fungal cultivation. The repeated-batch fungal fermentation enhanced lipid accumulation, which led to a final lipid concentration of 3.16 g/L on the digestate with 10% dry matter. The mass and energy balance analysis further indicates that the digestate had enough water for the process uses and the biogas energy was able to balance the needs of individual unit operations. A fresh-water-free and energy-positive process of lipid production from anaerobic digestate was achieved by integrating anaerobic digestion and fungal fermentation. The integration addresses the issues that both biofuel industry and waste management encounter-high water and energy demand of biofuel precursor production and few digestate utilization approaches of organic waste treatment.

Microbial communities in liquid and fiber fractions of food waste digestates are differentially resistant to inhibition by ammonia.

PubMed

Peng, Wei; Lü, Fan; Shao, Liming; He, Pinjing

2015-04-01

The effect of different concentrations of ammonia (1.0-7.0 g/L) during mesophilic anaerobic digestion with fiber or liquid digestate as inoculum was examined. Evolution of microbial community within fiber and liquid digestates was quantitatively assessed by the intact lipid analysis methods and qualitatively by DNA fingerprint methods in order to determine their resistance to ammonia inhibition. The results showed that an increased level of total ammonia nitrogen prolonged the lag phase of fiber digestates while reduced the metabolic rate of liquid digestates. Fiber digestates had 19.6-50.9-fold higher concentrations of phospholipid fatty acids (PLFA) compared to liquid digestates, whereas concentrations of phospholipid ether lipids (PLEL) in the fiber digestates were only 2.91-17.6-fold higher compared to liquid digestates. Although the cell concentration in liquid fraction was far lower than that in the fiber one, the ammonia-resistant ability and the methanization efficiency of the liquid digestate was superior to the fiber digestate. The bacterial profiles were affected more by the type of digestate inoculum compared to the concentration of ammonia. Principal component analysis indicated that the lipids technique was superior to the DNA technique for bacterial quantification but detected less archaeal diversity.

Evaluation of the pepsin digestibility assay for predicting amino acid digestibility of meat and bone meals.

PubMed

Davis, T M; Parsons, C M; Utterback, P L; Kirstein, D

2015-05-01

Sixteen meat and bone meal (MBM) samples were obtained and selected from various company plants to provide a wide range in pepsin nitrogen digestibility values. Pepsin digestibility was determined using either 0.02 or 0.002% pepsin. Amino acid (AA) digestibility of the 16 MBM samples was then determined using a precision-fed cecectomized rooster assay. The 0.02% pepsin digestibility values were numerically higher than the 0.002% pepsin values. The values varied from 77 to 93% for 0.02% pepsin and from 67 to 91% for 0.002% pepsin. The rooster AA digestibility results showed a wide range of values among MBM samples mostly due to the 4 samples having lowest and highest AA digestibility. A precision-fed broiler chick ileal AA digestibility assay confirmed that there were large differences in AA digestibility among the MBM samples having the lowest and highest rooster digestibility values. Correlation analyses between pepsin and AA digestibility values showed that the correlation values (r) were highly significant (P < 0.0001) for all AA when all 16 MBM samples were included in the analysis. However, when the MBM samples with the 2 lowest and the 2 highest rooster digestibility values were not included in the correlation analyses, the correlation coefficient values (r) were generally very low and not significant (P > 0.05). The results indicated that the pepsin nitrogen digestibility assay is only useful for detecting large differences in AA digestibility among MBM. There also was no advantage for using 0.02 versus 0.002% pepsin. © 2015 Poultry Science Association Inc.

Improving off-line accelerated tryptic digestion. Towards fast-lane proteolysis of complex biological samples.

PubMed

Vukovic, Jadranka; Loftheim, Håvard; Winther, Bjørn; Reubsaet, J Léon E

2008-06-27

Off-line digestion of proteins using immobilized trypsin beads is studied with respect to the format of the digestion reactor, the digestion conditions, the comparison with in-solution digestion and its use in complex biological samples. The use of the filter vial as the most appropriate digestion reactor enables simple, efficient and easy-to-handle off-line digestion of the proteins on trypsin beads. It was shown that complex proteins like bovine serum albumin (BSA) need much longer time (89 min) and elevated temperature (37 degrees C) to be digested to an acceptable level compared to smaller proteins like cytochrome c (5 min, room temperature). Comparing the BSA digestion using immobilized trypsin beads with conventional in-solution digestion (overnight at 37 degrees C), it was shown that comparable results were obtained with respect to sequence coverage (>90%) and amount of missed cleavages (in both cases around 20 peptides with 1 or 2 missed cleavages were detected). However, the digestion using immobilized trypsin beads was considerable less time consuming. Good reproducibility and signal intensities were obtained for the digestion products of BSA in a complex urine sample. In addition to this, peptide products of proteins typically present in urine were identified.

Biogas from mesophilic anaerobic digestion of cow dung using gelatin as additive

NASA Astrophysics Data System (ADS)

Salam, Bodius; Rahman, Md Mizanur; Sikder, Md Asif R.; Islam, Majedul

2017-06-01

A research work was conducted to investigate the enhanced production ability of biogas from mesophilic anaerobic digestions of cow dung (CD) using gelatin as additive. Five laboratory scale digesters were constructed to digest cow dung, where one set up was used for digestion of cow dung without additive and the other set up were used for digestion with additive. Gelatin additive was added in the slurry of amount 0.29, 0,57, 0.85 and 1.14% (wt.). The digesters were made of glass conical flask of 1-liter capacity each. Cow dung was used 335 gm and water was used 365 gm in each experiment. In the slurry, total solid content was maintained 8% (wt.) for all the observations. The digesters were fed on batch basis. The digesters were operated at ambient temperatures of 26 - 35°C. The total gas yield was obtained about 14.4 L/kg CD for digestion without additive and about 65% more biogas for digestion with 0.29% gelatin additive. The retention time for digestion without additive was 38 days and with additive retention time varied between 24 and 52 days.

Oxidation and ozonation of waste activated sludge.

PubMed

Mines, Richard O; Northenor, C Brett; Murchison, Mitchell

2008-05-01

In this bench-scale study, the treatment of waste activated sludge (WAS) was evaluated using aerobic digestion and ozonation. Two, 2-L batch digesters, one aerated and the other one ozonated, were operated for 30 days in each phase of the study. The aerated digester simulated the aerobic digestion process and served as control to the ozonated digester. In Phase I, the aerated digester was supplied 810 mg O(2) min(- 1), whereas, the ozonated digester was supplied 0.88 mg O(3) min(- 1). In Phase II, the oxygenation rate to the aerobic digester was increased to 1,200 mg O(2) min(- 1) while the ozonation rate was reduced to 0.44 mg O(3) min(- 1). Ozone was more effective than air at oxidizing and reducing both total solids (TS) and volatile solids (VS) in the WAS. TS removals of 50% and 56% were observed for the ozonated digester versus TS removals of 23% and 35% for the aerated digester. VS removals of 40% and 42% were observed for the aerobic digester versus 57% and 74% for the ozonated digester. Aerobic digestion barely met the 38% reduction in VS required by the U.S. Environmental Protection Agency (EPA). The degradation rate constant (K(d)) based on degradable TS for the ozonated digester varied from 0.082 to 0.11 days(- 1) and from 0.067 to 0.09 days(- 1) for the aerobic digester. Total chemical oxygen demand (TCOD) removal in the aerobic digester increased from 30% to 40% from Phase I to Phase II. TCOD removal increased slightly from 57% to 58% in the ozonated digester from Phase I to Phase II. Soluble chemical oxygen demand (SCOD) concentrations in the sludge supernatant increased with digestion time, especially in the ozonated digester. Approximately 0.12 to 0.22 mg SCOD was produced per mg of TS destroyed during ozonation. The specific oxygen uptake rate (SOUR) was consistently below the EPA standard of 1.5 mg O(2) per hr per g TS, indicating that the sludge was well stabilized. The average quantity of oxygen required during aerobic digestion was 1.53 g O(2) per g of TS destroyed. Actual ozone consumption rates were 0.57 mg O(3) per mg TS destroyed and 1.09 mg O(3) per mg TS destroyed for Phase II and Phase I, respectively.

Peptide profiling and the bioactivity character of yogurt in the simulated gastrointestinal digestion.

PubMed

Jin, Yan; Yu, Yang; Qi, Yanxia; Wang, Fangjun; Yan, Jiaze; Zou, Hanfa

2016-06-01

This study investigated the relationship between peptide profiles and the bioactivity character of yogurt in simulated gastrointestinal trials. A total of 250, 434 and 466 peptides were identified by LC-MS/MS analyses of yogurt, gastric digest and pancreatic digest. Forty peptides of yogurt survived in gastrointestinal digestion. κ-CN and β-CN contributed the diversity of peptides during the fermentation process and gastrointestinal digestion, respectively. The favorite of κ-CN by lactic acid bacteria complemented gut digestion by hydrolyzing κ-CN, the low abundance milk proteins. The potential bioactivities were evaluated by in vitro ACE and DPP-IV inhibition assays. The ACE inhibition rate of the pancreatic digests was ~4 - and ~2 - fold greater than that of yogurt and the gastric digests. The ACE inhibitory peptides generated during gastrointestinal digestion improved the ACE inhibitory activity of the gastric and pancreatic digests. The DPP-IV inhibition rate of the pancreatic digest was ~6 - and ~3 - fold greater than that of yogurt and the gastric digest. The numbers of potential DPP-IV inhibitory peptides were positively correlated to the DPP-IV inhibitory activity of the gastric and pancreatic digests. The present study describes the characters and bioactivities of peptides from yogurt in a simulated gastrointestinal digestion. The number of peptides identified from yogurt and gastrointestinal digests by LC-MS/MS increased in the simulated gastrointestinal trials. The in vitro ACE and DPP-IV inhibition bioactivities revealed that the bioactivity of yogurt was enhanced during gastrointestinal digestion. The correlation between peptides and bioactivity in vitro indicated that not only the peptides amount but also the proportion of peptides with high bioactivities contributed to increased bioactivity during gastrointestinal digestion. The study of peptides identified from yogurt and digests revealed that the number of released peptides was not determined by the abundance of the parent proteins but by whether the enzymes favored the protein. In summary, peptide profiling and bioactivities of yogurt in simulated gastrointestinal digestion helped to elucidate the health benefits of yogurt peptides. The results further revealed that pre-digestion of milk by lactic acid bacteria are complementary to generate bioactive peptides and to provide particular yogurt functions. Copyright © 2016 Elsevier B.V. All rights reserved.

Effect of temperature on methane production from field-scale anaerobic digesters treating dairy manure.

PubMed

Arikan, Osman A; Mulbry, Walter; Lansing, Stephanie

2015-09-01

Temperature is a critical factor affecting anaerobic digestion because it influences both system heating requirements and methane production. Temperatures of 35-37°C are typically suggested for manure digestion. In temperate climates, digesters require a considerable amount of additional heat energy to maintain temperatures at these levels. In this study, the effects of lower digestion temperatures (22 and 28°C), on the methane production from dairy digesters were evaluated and compared with 35°C using duplicate replicates of field-scale (FS) digesters with a 17-day hydraulic retention time. After acclimation, the FS digesters were operated for 12weeks using solids-separated manure at an organic loading rate (OLR) of 1.4kgVSm(-3)d(-1) and then for 8weeks using separated manure amended with manure solids at an OLR of 2.6kgVSm(-3)d(-1). Methane production values of the FS digesters at 22 and 28°C were about 70% and 87%, respectively, of the values from FS digesters at 35°C. The results suggest that anaerobic digesters treating dairy manure at 28°C were nearly as efficient as digesters operated at 35°C, with 70% of total methane achievable at 22°C. These results are relevant to small farms interested in anaerobic digestion for methane reduction without heat recovery from generators or for methane recovery from covered lagoon digesters. Copyright © 2015 Elsevier Ltd. All rights reserved.

Anaerobic digestion of chicken feather with swine manure or slaughterhouse sludge for biogas production.

PubMed

Xia, Yun; Massé, Daniel I; McAllister, Tim A; Beaulieu, Carole; Ungerfeld, Emilio

2012-03-01

Biogas production from anaerobic digestion of chicken feathers with swine manure or slaughterhouse sludge was assessed in two separate experiments. Ground feathers without any pre-treatment were added to 42-L digesters inoculated with swine manure or slaughterhouse sludge, representing 37% and 23% of total solids, respectively and incubated at 25°C in batch mode. Compared to the control without feather addition, total CH(4) production increased by 130% (P<0.001) and 110% (P=0.09) in the swine manure and the slaughterhouse sludge digesters, respectively. Mixed liquor NH(4)N concentration increased (P<0.001) from 4.8 and 3.1g/L at the beginning of the digestion to 6.9 and 3.5 g/L at the end of digestion in the swine manure and the slaughterhouse sludge digesters, respectively. The fraction of proteolytic microorganisms increased (P<0.001) during the digestion from 12.5% to 14.5% and 11.3% to 13.0% in the swine manure and the slaughterhouse sludge digesters with feather addition, respectively, but decreased in the controls. These results are reflective of feather digestion. Feather addition did not affect CH(4) yields of the swine manure digesters (P=0.082) and the slaughterhouse sludge digesters (P=0.21), indicating that feathers can be digested together with swine manure or slaughterhouse sludge without negatively affecting the digestion of swine manure and slaughterhouse sludge. Crown Copyright © 2011. Published by Elsevier Ltd. All rights reserved.

Application of an in vitro digestion model for 210Po bioaccessibility assessment in seafood.

PubMed

Roselli, Carla; Desideri, Donatella; Feduzi, Laura; Ugolini, Lucia; Meli, Maria Assunta

2017-12-01

This study aims to investigate the bioaccessibility of 210 Po in seafood and the impact of food preparation on this radionuclide. Polonium bioaccessibility is the fraction of 210 Po mobilised from food matrices into digestive extractants when applying an in vitro digestion model. The degree of bioaccessibility of 210 Po in food has important implications for estimating ingestion doses from this radionuclide. The simulation of gastrointestinal digestion was divided into three stages through the use of synthetic saliva, gastric and bile-pancreas solutions. Following pre-treatment with a saliva solution, raw and cooked seafood samples underwent one of the following treatments: (a) simulated gastric digestion only or (b) simulated complete gastrointestinal digestion (gastric digestion followed by bile-pancreas digestion). No significant difference (P > 0.05) in 210 Po mobility was found between samples that underwent gastric digestion compared to those that only underwent gastro-intestinal digestion. However, a significant difference (P < 0.05) in 210 Po bioaccessibility was found between raw and cooked seafood undergoing both gastric and gastro-intestinal digestion.

Improved biogas production from food waste by co-digestion with de-oiled grease trap waste.

PubMed

Wu, Li-Jie; Kobayashi, Takuro; Kuramochi, Hidetoshi; Li, Yu-You; Xu, Kai-Qin

2016-02-01

The objective of this study was to assess the feasibility of co-digesting food waste (FW) and de-oiled grease trap waste (GTW) to improve the biogas production. A lab-scale mesophilic digester (MD), a temperature-phased anaerobic digester (TPAD) and a TPAD with recycling (TPAD-R) were synchronously operated under mono-digestion (FW) and co-digestion (FW+de-oiled GTW). Co-digestion increased the biogas yield by 19% in the MD and TPAD-R, with a biogas yield of 0.60L/g VS added. Specific methanogenic activity in the TPAD-R was much higher than that in the MD. In addition to methane, hydrogen at a yield of approximately 1mol/mol hexose was produced in the TPAD-R. Alkalinity was consumed more in the co-digestion than in mono-digestion. Co-digestion resulted in more lipid accumulation in each digester. The MD favored the degradation of lipid and conversion of long-chain fatty acids more than the TPAD and TPAD-R. Copyright © 2015 Elsevier Ltd. All rights reserved.

Performance Evaluation of Existing Wedgewater and Vacuum-Assisted Bed Dewatering Systems

DTIC Science & Technology

1992-01-01

prior to dewatering by the wedgewater method. Of the 20 satisfied users, 11 preferred aerobic digestion , two employed anaerobic digestion, and seven...did not further process their sludge. Of the seven dissatisfied users, four used aerobic digestion and three employed anaerobic digestion. A meelic...queried, 11 employed aerobic digestion , two employed anaerobic digestion, and three did not process their sludge. Eight dissatisfied users employed

Aerobic sludge digestion under low dissolved oxygen concentrations.

PubMed

Arunachalam, RaviSankar; Shah, Hemant K; Ju, Lu-Kwang

2004-01-01

Low dissolved oxygen (DO) concentrations occur commonly in aerobic digesters treating thickened sludge, with benefits of smaller digester size, much reduced aeration cost, and higher digestion temperature (especially important for plants in colder areas). The effects of low DO concentrations on digestion kinetics were studied using the sludge from municipal wastewater treatment plants in Akron, Ohio, and Los Lunas, New Mexico. The experiments were conducted in both batch digestion and a mixed mode of continuous, fed-batch, and batch operations. The low DO condition was clearly advantageous in eliminating the need for pH control because of the simultaneous occurrence of nitrification and denitrification. However, when compared with fully aerobic (high DO) systems under constant pH control (rare in full-scale plants), low DO concentrations and a higher solids loading had a negative effect on the specific volatile solids (VS) digestion kinetics. Nonetheless, the overall (volumetric) digestion performance depends not only on the specific digestion kinetics, but also the solids concentration, pH, and digester temperature. All of the latter factors favor the low DO digestion of thickened sludge. The significant effect of temperature on low DO digestion was confirmed in the mixed-mode study with the Akron sludge. When compared with the well-known empirical correlation between VS reduction and the product (temperature x solids retention time), the experimental data followed the same trend, but were lower than the correlation predictions. The latter was attributed to the lower digestible VS in the Akron sludge, the slower digestion at low DO concentrations, or both. Through model simulation, the first-order decay constant (kd) was estimated as 0.004 h(-1) in the mixed-mode operations, much lower than those (0.011 to 0.029 h(-1)) obtained in batch digestion. The findings suggested that the interactions among sludges with different treatment ages may have a substantially negative effect on digestion kinetics. The use of multistage digesters, especially with small front-end reactors, may be advantageous in both "process" kinetics and "biological reaction" kinetics for sludge digestion.

Impact of a multicarbohydrase containing α-galactosidase and xylanase on ileal digestible energy, crude protein digestibility, and ileal amino acid digestibility in broiler chickens.

PubMed

Jasek, A; Latham, R E; Mañón, A; Llamas-Moya, S; Adhikari, R; Poureslami, R; Lee, J T

2018-06-08

Exogenous enzymatic supplementation of poultry feeds, including α-galactosidase and xylanase, has been shown to increase metabolically available energy, although little information has been published on the impact on amino acid digestibility. An experiment was conducted to investigate a multicarbohydrase containing α-galactosidase and xylanase on amino acid digestibility, ileal digestible energy (IDE), and CP in male broiler chicks. The experiment was a 2 × 2 (diet × enzyme) factorial arrangement with 15 replicates of 8 male broilers per replicate raised for 21 d in a battery setting. The 2 dietary treatments included a positive control (PC) and a negative control (NC) diet formulated to contain 2.5% less calculated AME and digestible amino acids. Each of these diets was fed with and without enzyme. Broilers were fed a starter diet from 0-14 d (crumble) and a grower from 14-21 d (pellet). Birds were sampled on day 21 to determine ileal amino acid digestibility, IDE, and CP digestibility. Titanium dioxide (TiO2) was used as an indigestible marker for the determination of digestibility coefficients. Total ileal amino acid digestibility was increased (P = 0.008) by 3.80% with the inclusion of enzyme. Methionine and lysine digestibility was improved (P < 0.05) with the inclusion of enzyme by 3.37% and 2.61%, respectively. Enzyme inclusion increased (P = 0.001) cysteine digestibility by 9.3%. Diet-influenced ileal amino acid digestibility with tryptophan, threonine, isoleucine, and valine digestibility being increased (P < 0.05) in the PC when compared to the NC. IDE was decreased (P = 0.037) in broilers fed the NC diet by 100 kcal/kg feed when compared to broilers fed the PC diet. Enzyme inclusion increased (P = 0.047) IDE value by 90 kcal/kg. Crude protein digestibility was not influenced by diet; however, similar improvements in CP digestibility with enzyme inclusion were observed as with energy. These data support the benefits of a multicarbohydrase containing α-galactosidase and xylanase inclusion to improve nutrient and ileal amino acid digestibility across multiple dietary nutrient profiles.

Engineering Digestion: Multiscale Processes of Food Digestion.

PubMed

Bornhorst, Gail M; Gouseti, Ourania; Wickham, Martin S J; Bakalis, Serafim

2016-03-01

Food digestion is a complex, multiscale process that has recently become of interest to the food industry due to the developing links between food and health or disease. Food digestion can be studied by using either in vitro or in vivo models, each having certain advantages or disadvantages. The recent interest in food digestion has resulted in a large number of studies in this area, yet few have provided an in-depth, quantitative description of digestion processes. To provide a framework to develop these quantitative comparisons, a summary is given here between digestion processes and parallel unit operations in the food and chemical industry. Characterization parameters and phenomena are suggested for each step of digestion. In addition to the quantitative characterization of digestion processes, the multiscale aspect of digestion must also be considered. In both food systems and the gastrointestinal tract, multiple length scales are involved in food breakdown, mixing, absorption. These different length scales influence digestion processes independently as well as through interrelated mechanisms. To facilitate optimized development of functional food products, a multiscale, engineering approach may be taken to describe food digestion processes. A framework for this approach is described in this review, as well as examples that demonstrate the importance of process characterization as well as the multiple, interrelated length scales in the digestion process. © 2016 Institute of Food Technologists®

Foaming phenomenon in bench-scale anaerobic digesters.

PubMed

Siebels, Amanda M; Long, Sharon C

2013-04-01

The Madison Metropolitan Sewerage District (The District) in Madison, Wisconsin has been experiencing seasonal foaming in their anaerobic biosolids digesters, which has occurred from mid-November to late June for the past few years. The exact cause(s) of foaming is unknown. Previous research findings are unclear as to whether applications of advanced anaerobic digestion processes reduce the foaming potential of digesters. The object of this study was to investigate how configurations of thermophilic and acid phase-thermophilic anaerobic digestion would affect foaming at the bench-scale level compared to single stage mesophilic digestion for The District. Bench-scale anaerobic digesters were fed with a 4 to 4.5% by dry weight of solids content blend of waste activated sludge (WAS) and primary sludge from The District. Foaming potential was monitored using Alka-Seltzer and aeration foaming tests. The bench-scale acid phase-thermophilic digester had a higher foaming potential than the bench-scale mesophilic digester. These results indicate that higher temperatures increase the foaming potential of the bench-scale anaerobic digesters. The bench-scale acid phase-thermophilic digesters had a greater percent (approximately 5 to 10%) volatile solids destruction and a greater percent (approximately 5 to 10%) total solids destruction when compared to the bench-scale mesophilic digester. Overall, for the full-scale foaming experienced by The District, it appears that adding an acid phase or switching to thermophilic digestion would not alleviate The District's foaming issues.

Inclusion of Digestible Surfactants in Solid SMEDDS Formulation Removes Lag Time and Influences the Formation of Structured Particles During Digestion.

PubMed

Vithani, Kapilkumar; Hawley, Adrian; Jannin, Vincent; Pouton, Colin; Boyd, Ben J

2017-05-01

Solid self-microemulsifying drug delivery systems (SMEDDS) have received considerable attention in recent times attempting to overcome the drawbacks of liquid SMEDDS. Earlier literature reports on solid SMEDDS have focussed on formulation development; however, the digestibility and propensity for self-assembly of the digested components with endogenous bile salts and phospholipids are unknown. Therefore, as a starting point, previously reported solid SMEDDS containing Gelucire® 44/14 (GEL) and the non-digestible surfactants, Vitamin E TPGS (TPGS) and Lutrol® F 127 (F 127), were prepared, and their dispersion and digestion behaviours were studied using an in vitro lipolysis model, coupled with small-angle X-ray scattering (SAXS) to determine the formed colloidal structures during digestion in real time. GEL alone was digested (89%) and formed a lamellar phase (Lα). When surfactants were added at a 40:60% w/w lipid to surfactants ratio, digestion was inhibited with a significant lag time being evident. However, increasing the fraction of GEL to 50% w/w enabled digestion with reduced lag time. The substitution of the non-digestible surfactants with digestible surfactants, sucrose esters S-1670 (S-1670) and Span® 60 (S-60), eliminated the digestion lag time, and the formation of colloidal structures was more similar to that of GEL alone.

High Hydrostatic Pressure Pretreatment of Whey Protein Isolates Improves Their Digestibility and Antioxidant Capacity

PubMed Central

Iskandar, Michèle M.; Lands, Larry C.; Sabally, Kebba; Azadi, Behnam; Meehan, Brian; Mawji, Nadir; Skinner, Cameron D.; Kubow, Stan

2015-01-01

Whey proteins have well-established antioxidant and anti-inflammatory bioactivities. High hydrostatic pressure processing of whey protein isolates increases their in vitro digestibility resulting in enhanced antioxidant and anti-inflammatory effects. This study compared the effects of different digestion protocols on the digestibility of pressurized (pWPI) and native (nWPI) whey protein isolates and the antioxidant and anti-inflammatory properties of the hydrolysates. The pepsin-pancreatin digestion protocol was modified to better simulate human digestion by adjusting temperature and pH conditions, incubation times, enzymes utilized, enzyme-to-substrate ratio and ultrafiltration membrane molecular weight cut-off. pWPI showed a significantly greater proteolysis rate and rate of peptide appearance regardless of digestion protocol. Both digestion methods generated a greater relative abundance of eluting peptides and the appearance of new peptide peaks in association with pWPI digestion in comparison to nWPI hydrolysates. Hydrolysates of pWPI from both digestion conditions showed enhanced ferric-reducing antioxidant power relative to nWPI hydrolysates. Likewise, pWPI hydrolysates from both digestion protocols showed similar enhanced antioxidant and anti-inflammatory effects in a respiratory epithelial cell line as compared to nWPI hydrolysates. These findings indicate that regardless of considerable variations of in vitro digestion protocols, pressurization of WPI leads to more efficient digestion that improves its antioxidant and anti-inflammatory properties. PMID:28231198

High Hydrostatic Pressure Pretreatment of Whey Protein Isolates Improves Their Digestibility and Antioxidant Capacity.

PubMed

Iskandar, Michèle M; Lands, Larry C; Sabally, Kebba; Azadi, Behnam; Meehan, Brian; Mawji, Nadir; Skinner, Cameron D; Kubow, Stan

2015-05-28

Whey proteins have well-established antioxidant and anti-inflammatory bioactivities. High hydrostatic pressure processing of whey protein isolates increases their in vitro digestibility resulting in enhanced antioxidant and anti-inflammatory effects. This study compared the effects of different digestion protocols on the digestibility of pressurized (pWPI) and native (nWPI) whey protein isolates and the antioxidant and anti-inflammatory properties of the hydrolysates. The pepsin-pancreatin digestion protocol was modified to better simulate human digestion by adjusting temperature and pH conditions, incubation times, enzymes utilized, enzyme-to-substrate ratio and ultrafiltration membrane molecular weight cut-off. pWPI showed a significantly greater proteolysis rate and rate of peptide appearance regardless of digestion protocol. Both digestion methods generated a greater relative abundance of eluting peptides and the appearance of new peptide peaks in association with pWPI digestion in comparison to nWPI hydrolysates. Hydrolysates of pWPI from both digestion conditions showed enhanced ferric-reducing antioxidant power relative to nWPI hydrolysates. Likewise, pWPI hydrolysates from both digestion protocols showed similar enhanced antioxidant and anti-inflammatory effects in a respiratory epithelial cell line as compared to nWPI hydrolysates. These findings indicate that regardless of considerable variations of in vitro digestion protocols, pressurization of WPI leads to more efficient digestion that improves its antioxidant and anti-inflammatory properties.

Determination of metal concentrations in certified plastic reference materials after small-size autoclave and microwave-assisted digestion followed with inductively coupled plasma optical emission spectrometry

NASA Astrophysics Data System (ADS)

Lehtimäki, Esa; Väisänen, Ari

2017-01-01

The digestion methods for the determination of As, Cd, Cr, Pb, Sb, Sn and Zn concentrations in plastic samples using microwave-assisted digestion (MW-AD) and small-size autoclave digestion was developed. The certified polyethylene, polypropylene, polyvinyl chloride and acrylonitrile butadiene styrene certified reference materials were used in order to find digestion method working properly for several sample matrices. Efficiency of the digestion methods was evaluated by analyzing the residual carbon in digests by TOC analyzer. MW-AD using a mixture of 7 mL of HNO3 and 3 mL of H2O2 as a digestion solution resulted in excellent recoveries for As, Cd, Pb, Sb and Zn, and were in the range of 92-107% for all the analytes except Pb in polyethylene material. Autoclave digestion using 5 mL of concentrated HNO3 as a digestion solution resulted in similar recoveries with the exception of a higher As recovery (98%). Tin recovery resulted in low level after both MW-AD and autoclave digestion. Autoclave digestion was further developed resulting in a partially open two-step digestion process especially for the determination of Sn and Cr. The method resulted in higher recoveries of Sn and Cr (87 and 76%) but with the lower concentration of easily volatile As, Cd and Sb.

Actinidin enhances protein digestion in the small intestine as assessed using an in vitro digestion model.

PubMed

Kaur, Lovedeep; Rutherfurd, Shane M; Moughan, Paul J; Drummond, Lynley; Boland, Mike J

2010-04-28

This paper describes an in vitro study that tests the proposition that actinidin from green kiwifruit influences the digestion of proteins in the small intestine. Different food proteins, from sources including soy, meat, milk, and cereals, were incubated in the presence or absence of green kiwifruit extract (containing actinidin) using a two-stage in vitro digestion system consisting of an incubation with pepsin at stomach pH (simulating gastric digestion) and then with added pancreatin at small intestinal pH, simulating upper tract digestion in humans. The digests from the small intestinal stage (following the gastric digestion phase) were subjected to gel electrophoresis (SDS-PAGE) to assess loss of intact protein and development of large peptides during the in vitro simulated digestion. Kiwifruit extract influenced the digestion patterns of all of the proteins to various extents. For some proteins, actinidin had little impact on digestion. However, for other proteins, the presence of kiwifruit extract resulted in a substantially greater loss of intact protein and different peptide patterns from those seen after digestion with pepsin and pancreatin alone. In particular, enhanced digestion of whey protein isolate, zein, gluten, and gliadin was observed. In addition, reverse-phase HPLC (RP-HPLC) analysis showed that a 2.5 h incubation of sodium caseinate with kiwifruit extract alone resulted in approximately 45% loss of intact protein.

Anaerobic Co-digestion for Enhanced Renewable Energy and Green House Gas Emission Reduction

DOE Office of Scientific and Technical Information (OSTI.GOV)

Navaratnam, Navaneethan; Zitomer, Daniel

2012-05-01

The need to develop renewable energy is important for replacing fossil fuel, which is limited in quantity and also tends to increase in price over time. The addition of high strength organic wastes in municipal anaerobic digesters is growing and tends to increase renewable energy production. In addition, conversion of wastes to energy significantly reduces uncontrolled greenhouse gas emissions. Co-digestion of municipal sludge with any combination of wastes can result in synergistic, antagonistic or neutral outcomes. The objectives of this study were to identify potential co-digestates, determine synergistic, antagonistic and neutral effects, determine economic benefits, quantify performance of bench scalemore » co-digesters, identify influence of co-digestion on microbial communities and implement appropriate co-digestion, if warranted, after full-scale testing. A market study was used to identify promising co-digestates. Most promising wastes were determined by biochemical methane potential (BMP) and other testing followed by a simple economic analysis. Performance was investigated using bench-scale digesters receiving synthetic primary sludge with and without co-digestates. Denaturing gradient gel electrophoresis (DGGE) and quantitative polymerase chain reaction (qPCR) analyses were performed on the gene encoding the α subunit of methyl coenzyme M reductase (mcrA) to compare methanogen communities among the digesters. One significant band contributing to the greatest difference in banding patterns was excised, cloned, amplified and sequenced. Full- scale co-digestion was conducted using the most promising co-digestate at South Shore Wastewater Reclamation Facility (Oak Creek, WI). Over 80 wastes were identified from 54 facilities within 160 km of an existing municipal digester. A simple economic comparison identified the greatest benefits for seven co-digestates. Methane production rates of two co- digester systems increased by 105% and 66% in comparison to a control system. These increases were great than anticipated based on theoretical methane production from the additional chemical oxygen demand (COD) of the co-digestates. Co-digestion of the most promising wastes with primary sludge was estimated to generate enough electricity to power more than 2500 houses. Synergistic outcomes of co-digestion may be caused by chances in microbial community resulting in more rapid methane production rate and higher specific methanogenic activities of the biomass against acetate, propionate and H2 as substrates. The presence of Methanospirillum hungatei correlated to higher SMAs in the Co-Digester 1 system. In subsequent full-scale testing, acid whey in addition to primary sludge increased methane production by 16 %, biogas methane content by 5%, methane yield per VS destroyed by 9% ( from 650 to 710 L CH4 / kg VSdestroyed ) and volatile solids removal by 20%. Co-digestion is a promising technology to increase renewable energy production and convert municipal digesters into regional renewable energy facilities.« less

Optimization of microwave digestion for mercury determination in marine biological samples by cold vapour atomic absorption spectrometry.

PubMed

Cardellicchio, Nicola; Di Leo, Antonella; Giandomenico, Santina; Santoro, Stefania

2006-01-01

Optimization of acid digestion method for mercury determination in marine biological samples (dolphin liver, fish and mussel tissues) using a closed vessel microwave sample preparation is presented. Five digestion procedures with different acid mixtures were investigated: the best results were obtained when the microwave-assisted digestion was based on sample dissolution with HNO3-H2SO4-K2Cr2O7 mixture. A comparison between microwave digestion and conventional reflux digestion shows there are considerable losses of mercury in the open digestion system. The microwave digestion method has been tested satisfactorily using two certified reference materials. Analytical results show a good agreement with certified values. The microwave digestion proved to be a reliable and rapid method for decomposition of biological samples in mercury determination.

A critical review on the interaction of substrate nutrient balance and microbial community structure and function in anaerobic co-digestion.

PubMed

Xu, Rong; Zhang, Kai; Liu, Pu; Khan, Aman; Xiong, Jian; Tian, Fake; Li, Xiangkai

2018-01-01

Anaerobic co-digestion generally results in a higher yield of biogas than mono-digestion, hence co-digestion has become a topic of general interest in recent studies of anaerobic digestion. Compared with mono-digestion, co-digestion utilizes multiple substrates. The balance of substrate nutrient in co-digestion comprises better adjustments of C/N ratio, pH, moisture, trace elements, and dilution of toxic substances. All of these changes could result in positive shifts in microbial community structure and function in the digestion processes and consequent augmentation of biogas production. Nevertheless, there have been few reviews on the interaction of nutrient and microbial community in co-digestions. The objective of this review is to investigate recent achievements and perspectives on the interaction of substrate nutrient balance and microbial community structure and function. This may provide valuable information on the optimization of combinations of substrates and prediction of bioreactor performance. Copyright © 2017 Elsevier Ltd. All rights reserved.

Assessment of Two Solid Anaerobic Digestate Soil Amendments for Effects on Soil Quality and Biosolarization Efficacy.

PubMed

Fernández-Bayo, Jesús D; Achmon, Yigal; Harrold, Duff R; McCurry, Dlinka G; Hernandez, Katie; Dahlquist-Willard, Ruth M; Stapleton, James J; VanderGheynst, Jean S; Simmons, Christopher W

2017-05-03

Anaerobic digestion is an organic waste bioconversion process that produces biofuel and digestates. Digestates have potential to be applied as soil amendment to improve properties for crop production including phytonutrient content and pest load. Our objective was to assess the impact of solid anaerobic digestates on weed seed inactivation and soil quality upon soil biosolarization (a pest control technique that combines solar heating and amendment-induced microbial activity). Two solid digestates from thermophilic (TD) and mesophilic (MD) digesters were tested. The solarized TD-amended samples presented significantly higher mortality of Brassica nigra (71%, P = 0.032) than its equivalent incubated at room temperature. However, biosolarization with digestate amendment led to decreased weed seed mortality in certain treatments. The plant-available water, total C, and extractable P and K were significantly increased (P < 0.05) in the incubated amended soils. The results confirm the potential of digestates as beneficial soil amendments. Further studies are needed to elucidate the impacts of digestate stability on biosolarization efficacy and soil properties.

Sequential Anaerobic/Aerobic Digestion for Enhanced Carbon/Nitrogen Removal and Cake Odor Reduction.

PubMed

Ahmad, Muneer; Denee, Marco Abel; Jiang, Hao; Eskicioglu, Cigdem; Kadota, Paul; Gregonia, Theresa

2016-12-01

  Anaerobic digestion (AD) has been proven to be an effective process for the treatment of wastewater sludge. However, it produces high levels of ammonia in the digester effluent, which may jeopardize meeting stringent nutrient discharge limits. In this study, the effect of a sequential anaerobic/aerobic (AN/AERO) digestion and a single-stage conventional AN digestion (as control) was investigated on mixed (primary + secondary) sludge generated by the Annacis Island wastewater treatment plant (WWTP) (BC, Canada). An overall sludge retention time (SRT) of 22.5 days under three different scenarios was chosen based on the current operational SRT of the digesters at the Annacis Island WWTP. The steady state results have shown that sequential AN/AERO digestion configurations achieved up to 11% higher volatile solids (VS) removal and 72% lower ammonia generation over single-stage conventional AN digestion. Furthermore, sequential AN/AERO system also showed enhanced dewaterability, improved fecal coliform destruction and reduced digested cake odors over control digesters.

Comparative molecular cytogenetic analyses of a major tandemly repeated DNA family and retrotransposon sequences in cultivated jute Corchorus species (Malvaceae).

PubMed

Begum, Rabeya; Zakrzewski, Falk; Menzel, Gerhard; Weber, Beatrice; Alam, Sheikh Shamimul; Schmidt, Thomas

2013-07-01

The cultivated jute species Corchorus olitorius and Corchorus capsularis are important fibre crops. The analysis of repetitive DNA sequences, comprising a major part of plant genomes, has not been carried out in jute but is useful to investigate the long-range organization of chromosomes. The aim of this study was the identification of repetitive DNA sequences to facilitate comparative molecular and cytogenetic studies of two jute cultivars and to develop a fluorescent in situ hybridization (FISH) karyotype for chromosome identification. A plasmid library was generated from C. olitorius and C. capsularis with genomic restriction fragments of 100-500 bp, which was complemented by targeted cloning of satellite DNA by PCR. The diversity of the repetitive DNA families was analysed comparatively. The genomic abundance and chromosomal localization of different repeat classes were investigated by Southern analysis and FISH, respectively. The cytosine methylation of satellite arrays was studied by immunolabelling. Major satellite repeats and retrotransposons have been identified from C. olitorius and C. capsularis. The satellite family CoSat I forms two undermethylated species-specific subfamilies, while the long terminal repeat (LTR) retrotransposons CoRetro I and CoRetro II show similarity to the Metaviridea of plant retroelements. FISH karyotypes were developed by multicolour FISH using these repetitive DNA sequences in combination with 5S and 18S-5·8S-25S rRNA genes which enable the unequivocal chromosome discrimination in both jute species. The analysis of the structure and diversity of the repeated DNA is crucial for genome sequence annotation. The reference karyotypes will be useful for breeding of jute and provide the basis for karyotyping homeologous chromosomes of wild jute species to reveal the genetic and evolutionary relationship between cultivated and wild Corchorus species.

Digestive stimulant action of spices: a myth or reality?

PubMed

Platel, Kalpana; Srinivasan, K

2004-05-01

Spices have long been recognized for their digestive stimulant action. Several spices are also employed in medicinal preparations against digestive disorders in traditional and Indian systems of medicine. Earlier reports on the digestive stimulant action of spices are largely empirical; only in recent years, this beneficial attribute of spices has been authenticated in exhaustive animal studies. Animal studies have shown that many spices induce higher secretion of bile acids which play a vital role in fat digestion and absorption. When consumed through the diet also spices produce significant stimulation of the activities of pancreatic lipase, amylase and proteases. A few of them also have been shown to have beneficial effect on the terminal digestive enzymes of small intestinal mucosa. Concomitant with such a stimulation of either bile secretion or activity of digestive enzymes by these spices, leading to an accelerated digestion, a reduction in the food transit time in the gastrointestinal tract has also been shown. Thus, the digestive stimulant action of spices seems to be mediated through two possible modes: (i) by stimulating the liver to secrete bile rich in bile acids, components that are vital for fat digestion and absorption, and (ii) by a stimulation of enzyme activities that are responsible for digestion. This review highlights the available information on the influence of spices on the digestive secretions and enzymes.

Identification of a low digestibility δ-Conglutin in yellow lupin (Lupinus luteus L.) seed meal for atlantic salmon (Salmo salar L.) by coupling 2D-PAGE and mass spectrometry.

PubMed

Ogura, Takahiro; Hernández, Adrián; Aizawa, Tomoko; Ogihara, Jun; Sunairi, Michio; Alcaino, Javier; Salvo-Garrido, Haroldo; Maureira-Butler, Iván J

2013-01-01

The need of quality protein in the aquaculture sector has forced the incorporation of alternative plant proteins into feeding diets. However, most plant proteins show lower digestibility levels than fish meal proteins, especially in carnivorous fishes. Manipulation of protein content by plant breeding can improve the digestibility rate of plant proteins in fish, but the identification of low digestibility proteins is essential. A reduction of low digestibility proteins will not only increase feed efficiency, but also reduce water pollution. Little is known about specific digestible protein profiles and/or molecular identification of more bioavailable plant proteins in fish diets. In this study, we identified low digestibility L. luteus seed proteins using Atlantic salmon (Salmo salar) crude digestive enzymes in an in vitro assay. Low digestibility proteins were identified by comparing SDS-PAGE banding profiles of digested and non-digested lupin seed proteins. Gel image analysis detected a major 12 kDa protein band in both lupin meal and protein isolate digested products. The 12 kDa was confirmed by 2D-PAGE gels and the extracted protein was analyzed with an ion trap mass spectrometer in tandem mass mode. The MS/MS data showed that the 12 kDa low digestibility protein was a large chain δconglutin, a common seed storage protein of yellow lupin. Comparison of the protein band profiles between lupin meal and protein isolates showed that the isolatation process did not affect the low digestibility of the 12 kDa protein.

Identification of a Low Digestibility δ-Conglutin in Yellow Lupin (Lupinus luteus L.) Seed Meal for Atlantic Salmon (Salmo salar L.) by Coupling 2D-PAGE and Mass Spectrometry

PubMed Central

Ogura, Takahiro; Hernández, Adrián; Aizawa, Tomoko; Ogihara, Jun; Sunairi, Michio; Alcaino, Javier; Salvo-Garrido, Haroldo; Maureira-Butler, Iván J.

2013-01-01

The need of quality protein in the aquaculture sector has forced the incorporation of alternative plant proteins into feeding diets. However, most plant proteins show lower digestibility levels than fish meal proteins, especially in carnivorous fishes. Manipulation of protein content by plant breeding can improve the digestibility rate of plant proteins in fish, but the identification of low digestibility proteins is essential. A reduction of low digestibility proteins will not only increase feed efficiency, but also reduce water pollution. Little is known about specific digestible protein profiles and/or molecular identification of more bioavailable plant proteins in fish diets. In this study, we identified low digestibility L. luteus seed proteins using Atlantic salmon (Salmo salar) crude digestive enzymes in an in vitro assay. Low digestibility proteins were identified by comparing SDS-PAGE banding profiles of digested and non-digested lupin seed proteins. Gel image analysis detected a major 12 kDa protein band in both lupin meal and protein isolate digested products. The 12 kDa was confirmed by 2D-PAGE gels and the extracted protein was analyzed with an ion trap mass spectrometer in tandem mass mode. The MS/MS data showed that the 12 kDa low digestibility protein was a large chain δconglutin, a common seed storage protein of yellow lupin. Comparison of the protein band profiles between lupin meal and protein isolates showed that the isolatation process did not affect the low digestibility of the 12 kDa protein. PMID:24278278

Geriatric syndromes increased the nutritional risk in elderly cancer patients independently from tumour site and metastatic status. The ELCAPA-05 cohort study.

PubMed

Paillaud, E; Liuu, E; Laurent, M; Le Thuaut, A; Vincent, H; Raynaud-Simon, A; Bastuji-Garin, S; Tournigand, C; Caillet, P; Canoui-Poitrine, F

2014-04-01

We assessed the prevalence and risk factors of malnutrition in elderly cancer patients. We studied a prospective cohort of solid cancer patients aged ≥70 years at referral to two geriatric oncology clinics between 2007 and 2010. Nutrition was evaluated using the Mini-Nutritional Assessment (MNA) using validated cut-offs (<17: malnutrition, 17-23.5: at-risk for malnutrition). Patients with non-digestive tumours (breast, prostate, urinary tract) and with digestive (colorectal, upper digestive tract and liver) were analysed separately using multinomial logistic regression. Of 643 consecutive patients, 519 had available data (median age, 80; men, 48.2%; metastases, 46.3%; digestive cancer 47.8%). In non-digestive group, 13.3% had malnutrition versus 28.6% in digestive group. The link between metastasis and malnutrition was significantly higher in non-digestive group (adjusted odds ratio [ORa ], 25.25; 95%CI: 5.97-106.8) than in digestive group (ORa, 2.59; 1.08-6.24; p for heterogeneity = 0.04). Other factors independently associated with malnutrition were cognitive impairment (ORa MMMSE ≤ 24 versus > 24 in non-digestive group: 16.68; 4.89-56.90 and in digestive group: 3.93; 1.34-11.50), and depressed mood (ORa MiniGDS ≥1 versus <1 in non-digestive group: 11.11; 3.32-37.17 and in digestive group: 3.25; 1.29-8.15) and fall risk (ORa fall risk versus no fall risk in non-digestive group: 4.68; 1.77-12.37; in digestive group: 100% of malnourished patients were faller's). We highlighted, in elderly cancer patients, the high prevalence of malnutrition and that geriatrics syndromes (i.e. cognitive impairment, depressed mood and fall risk) were independent risk factors for malnutrition. Moreover, metastatic status was significantly much more strongly associated with malnutrition in non-digestive than digestive tumours. Copyright © 2013 Elsevier Ltd and European Society for Clinical Nutrition and Metabolism. All rights reserved.

Influence of resistant starch and slowly digestible starch on rice texture.

USDA-ARS?s Scientific Manuscript database

Rice, comprised mainly of starch, serves as a significant source of caloric energy world-wide, therefore differences in starch digestibility are important to human health. Rice starch consists of three forms based on digestibility, rapidly digestible starch (RDS), slowly digestible starch (SDS), and...

40 CFR 428.80 - Applicability; description of the wet digestion reclaimed rubber subcategory.

Code of Federal Regulations, 2010 CFR

2010-07-01

... digestion reclaimed rubber subcategory. 428.80 Section 428.80 Protection of Environment ENVIRONMENTAL... Wet Digestion Reclaimed Rubber Subcategory § 428.80 Applicability; description of the wet digestion... discharges resulting from the production of reclaimed rubber by use of the wet digestion process. ...

40 CFR 428.80 - Applicability; description of the wet digestion reclaimed rubber subcategory.

Code of Federal Regulations, 2011 CFR

2011-07-01

... digestion reclaimed rubber subcategory. 428.80 Section 428.80 Protection of Environment ENVIRONMENTAL... Wet Digestion Reclaimed Rubber Subcategory § 428.80 Applicability; description of the wet digestion... discharges resulting from the production of reclaimed rubber by use of the wet digestion process. ...

76 FR 29771 - National Institute of Diabetes and Digestive and Kidney Diseases; Notice of Closed Meetings

Federal Register 2010, 2011, 2012, 2013, 2014

2011-05-23

... Diabetes and Digestive and Kidney Diseases; Notice of Closed Meetings Pursuant to section 10(d) of the... Diabetes and Digestive and Kidney Diseases Special Emphasis Panel, NIDDK Interconnectivity Network. Date... and Digestive and Kidney Diseases Special Emphasis Panel, Digestive Diseases Program Projects. Date...

Phosphatidyl derivative of hydroxytyrosol. In vitro intestinal digestion, bioaccessibility, and its effect on antioxidant activity.

PubMed

Martin, Diana; Moran-Valero, Maria I; Casado, Víctor; Reglero, Guillermo; Torres, Carlos F

2014-10-08

Intestinal digestion of phosphatidyl derivatives of HT (PHT) and its bioaccessibility under in vitro conditions was performed. First, an in vitro intestinal digestion model for phospholipids was developed. The impact of digestion in the antioxidant ability of PHT was also assayed. PHT was progressively hydrolyzed to lyso-PHT. However, digestion was slower than the phospholipid control. Nevertheless, most hydrolysis products were found at the micellar phase fraction, meaning a high bioaccessibility. Either PHT or digested PHT showed lower antioxidant activity than HT. However, PHT improved its antioxidant ability after digestion, likely related to lyso-PHT. As a summary, the synthetic phosphatidyl derivative of HT as PHT is recognized by phospholipases during simulation of intestinal digestion, although less efficiently than analogous phospholipids. Nevertheless, taking into account the bioaccessibility and the antioxidant activity of digested PHT, the potential of carriers of HT under the form of phospholipids might be of interest.

Semi-continuous anaerobic co-digestion of thickened waste activated sludge and fat, oil and grease

DOE Office of Scientific and Technical Information (OSTI.GOV)

Wan Caixia; Zhou Quancheng; Fu Guiming

2011-08-15

Highlights: > Co-digestion of thickened waste activated sludge (TWAS) with fat, oil and grease (FOG). > Co-digestion of TWAS and FOG at 64% VS increased biogas production by 137%. > FOG addition ratio at 74% of total VS caused inhibition of the anaerobic digestion process. > Micronutrients addition did not significantly improve the biogas production and digestion stabilization. - Abstract: Co-digestion of thickened waste activated sludge (TWAS) and fat, oil and grease (FOG) was conducted semi-continuously under mesophilic conditions. The results showed that daily methane yield at the steady state was 598 L/kg VS{sub added} when TWAS and FOG (64%more » of total VS) were co-digested, which was 137% higher than that obtained from digestion of TWAS alone. The biogas composition was stabilized at a CH{sub 4} and CO{sub 2} content of 66.8% and 29.5%, respectively. Micronutrients added to co-digestion did not improve the biogas production and digestion stabilization. With a higher addition of FOG (74% of total VS), the digester initially failed but was slowly self-recovered; however, the methane yield was only about 50% of a healthy reactor with the same organic loading rate.« less

Survival rates of parasite eggs in sludge during aerobic and anaerobic digestion.

PubMed Central

Black, M I; Scarpino, P V; O'Donnell, C J; Meyer, K B; Jones, J V; Kaneshiro, E S

1982-01-01

The effects of mesothermic anaerobic or aerobic sludge digestion on survival of eggs from the roundworms Ascaris suum, toxocara canis, Trichuris vulpis, and Trichuris suis and from the rat tapeworm Hymenolepis diminuta were studied. Destruction of eggs throughout a 15-day treatment period, as well as their viabilities after reisolation, was analyzed. The laboratory model digesters used in this study were maintained at a 15-day retention schedule, partially simulating a continuously operating system. Ascaris eggs were destroyed in the anaerobic (23%) or aerobic (38%) digesters, and 11% Trichuris eggs were destroyed in the aerobic digesters. Trichuris eggs in anaerobic digesters and Toxocara eggs in either anaerobic or aerobic digesters were not destroyed. Destruction of eggs in digesters was correlated with the state of the eggs before subjection to the treatment processes; i.e., some Ascaris and Trichuris eggs were already embryonated in host intestinal contents or feces and hence past their most resistant stage. The viabilities of Ascaris and Toxocara eggs that survived the digestion processes were greater in anaerobically treated than in aerobically treated material. Eggs from Hymenolepis were nonviable before use in the experiments. However, they were more effectively destroyed in aerobic digesters than in anaerobic digesters. PMID:6891199

Benefits of supplementing an industrial waste anaerobic digester with energy crops for increased biogas production

DOE Office of Scientific and Technical Information (OSTI.GOV)

Nges, Ivo Achu, E-mail: Nges.Ivo_Achu@biotek.lu.se; Escobar, Federico; Fu Xinmei

2012-01-15

Highlights: Black-Right-Pointing-Pointer This study demonstrates the feasibility of co-digestion food industrial waste with energy crops. Black-Right-Pointing-Pointer Laboratory batch co-digestion led to improved methane yield and carbon to nitrogen ratio as compared to mono-digestion of industrial waste. Black-Right-Pointing-Pointer Co-digestion was also seen as a means of degrading energy crops with nutrients addition as crops are poor in nutrients. Black-Right-Pointing-Pointer Batch co-digestion methane yields were used to predict co-digestion methane yield in full scale operation. Black-Right-Pointing-Pointer It was concluded that co-digestion led an over all economically viable process and ensured a constant supply of feedstock. - Abstract: Currently, there is increasing competitionmore » for waste as feedstock for the growing number of biogas plants. This has led to fluctuation in feedstock supply and biogas plants being operated below maximum capacity. The feasibility of supplementing a protein/lipid-rich industrial waste (pig manure, slaughterhouse waste, food processing and poultry waste) mesophilic anaerobic digester with carbohydrate-rich energy crops (hemp, maize and triticale) was therefore studied in laboratory scale batch and continuous stirred tank reactors (CSTR) with a view to scale-up to a commercial biogas process. Co-digesting industrial waste and crops led to significant improvement in methane yield per ton of feedstock and carbon-to-nitrogen ratio as compared to digestion of the industrial waste alone. Biogas production from crops in combination with industrial waste also avoids the need for micronutrients normally required in crop digestion. The batch co-digestion methane yields were used to predict co-digestion methane yield in full scale operation. This was done based on the ratio of methane yields observed for laboratory batch and CSTR experiments compared to full scale CSTR digestion of industrial waste. The economy of crop-based biogas production is limited under Swedish conditions; therefore, adding crops to existing industrial waste digestion could be a viable alternative to ensure a constant/reliable supply of feedstock to the anaerobic digester.« less

Digestive diseases

MedlinePlus

... this page: //medlineplus.gov/ency/article/007447.htm Digestive diseases To use the sharing features on this page, please enable JavaScript. Digestive diseases are disorders of the digestive tract, which ...

DOE Office of Scientific and Technical Information (OSTI.GOV)

Bayr, S., E-mail: suvi.bayr@jyu.fi; Ojanperä, M.; Kaparaju, P.

Highlights: • Rendering wastes’ mono-digestion and co-digestion with potato pulp were studied. • CSTR process with OLR of 1.5 kg VS/m{sup 3} d, HRT of 50 d was unstable in mono-digestion. • Free NH{sub 3} inhibited mono-digestion of rendering wastes. • CSTR process with OLR of 1.5 kg VS/m{sup 3} d, HRT of 50 d was stable in co-digestion. • Co-digestion increased methane yield somewhat compared to mono-digestion. - Abstract: In this study, mono-digestion of rendering wastes and co-digestion of rendering wastes with potato pulp were studied for the first time in continuous stirred tank reactor (CSTR) experiments at 55more » °C. Rendering wastes have high protein and lipid contents and are considered good substrates for methane production. However, accumulation of digestion intermediate products viz., volatile fatty acids (VFAs), long chain fatty acids (LCFAs) and ammonia nitrogen (NH{sub 4}-N and/or free NH{sub 3}) can cause process imbalance during the digestion. Mono-digestion of rendering wastes at an organic loading rate (OLR) of 1.5 kg volatile solids (VS)/m{sup 3} d and hydraulic retention time (HRT) of 50 d was unstable and resulted in methane yields of 450 dm{sup 3}/kg VS{sub fed}. On the other hand, co-digestion of rendering wastes with potato pulp (60% wet weight, WW) at the same OLR and HRT improved the process stability and increased methane yields (500–680 dm{sup 3}/kg VS{sub fed}). Thus, it can be concluded that co-digestion of rendering wastes with potato pulp could improve the process stability and methane yields from these difficult to treat industrial waste materials.« less

Intestinal digestibility of long-chain fatty acids in lactating dairy cows: A meta-analysis and meta-regression.

PubMed

Boerman, J P; Firkins, J L; St-Pierre, N R; Lock, A L

2015-12-01

The objective of this analysis was to examine the intestinal digestibility of individual long-chain fatty acids (FA) in lactating dairy cows. Available data were collated from 15 publications containing 61 treatments, which reported total and individual FA duodenal flows and calculations of intestinal digestibility. All studies involved lactating dairy cows, and estimates of digestibility were based on measurements either between the duodenum and ileum (18 treatments) or between the duodenum and feces (43 treatments). Fatty acid digestibility was calculated for C16:0, C18:0, C18:1 (cis and trans isomers), C18:2, and C18:3. Digestibility of C18:0 was lower than for C18:1 and C18:3, with no difference in digestibility between saturated FA (C16:0 and C18:0). We weighted the studies by the reciprocal of the variance to generate best-fit equations to predict individual FA digestibility based on duodenal flow of FA and dietary independent variables. The flow of C18:0 negatively affected the digestibility of C18:0 and was also included in the best-fit equations for all other 18-carbon FA using duodenal flow characteristics. The type of fat supplemented had an effect on digestibility of individual FA, with whole seeds having reduced digestibility. Our meta-analysis results showed minimal differences in the digestibility of individual FA. However, C18:0 flow through the duodenum had a negative effect on the digestibility of several individual FA, with the largest negative effect on C18:0 digestibility. The mechanisms that reduce C18:0 absorption at high concentrations are unknown and warrant further investigation. Copyright © 2015 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Antioxidant and antidiabetic properties of tartary buckwheat rice flavonoids after in vitro digestion*

PubMed Central

Bao, Tao; Wang, Ye; Li, Yu-ting; Gowd, Vemana; Niu, Xin-he; Yang, Hai-ying; Chen, Li-shui; Chen, Wei; Sun, Chong-de

2016-01-01

Oxidative stress and diabetes have a tendency to alter protein, lipid, and DNA moieties. One of the strategic methods used to reduce diabetes-associated oxidative stress is to inhibit the carbohydrate-digesting enzymes, thereby decreasing gastrointestinal glucose production. Plant-derived natural antioxidant molecules are considered a therapeutic tool in the treatment of oxidative stress and diabetes. The objective of this study was to identify tartary buckwheat rice flavonoids and evaluate the effect of in vitro digestion on their antioxidant and antidiabetic properties. High performance liquid chromatography (HPLC) analysis indicated the presence of rutin as a major component and quercitrin as a minor component of both digested and non-digested flavonoids. Both extracts showed a significant antioxidant capacity, but digested flavonoids showed reduced activity compared to non-digested. There were some decreases of the antioxidant activities (2,2'-azino-bis-(3-ethylbenzthiazoline-6-sulfonic acid) diammonium salt (ABTS), 2,2-diphenyl-1-picrylhydrazy (DPPH) radical, and ferric reducing antioxidant power (FRAP)) of digested tartary buckwheat rice flavonoids compared with non-digested. Flavonoids from both groups significantly inhibited reactive oxygen species (ROS) production and α-glucosidase activity. Both digested and non-digested flavonoids markedly increased glucose consumption and glycogen content in HepG2 cells. Tartary buckwheat rice flavonoids showed appreciable antioxidant and antidiabetic properties, even after digestion. Tartary buckwheat rice appears to be a promising functional food with potent antioxidant and antidiabetic properties. PMID:27921399

Solid anaerobic digestion batch with liquid digestate recirculation and wet anaerobic digestion of organic waste: Comparison of system performances and identification of microbial guilds.

PubMed

Di Maria, Francesco; Barratta, Martino; Bianconi, Francesco; Placidi, Pisana; Passeri, Daniele

2017-01-01

Solid anaerobic digestion batch (SADB) with liquid digestate recirculation and wet anaerobic digestion of organic waste were experimentally investigated. SADB was operated at an organic loading rate (OLR) of 4.55kgVS/m 3 day, generating about 252NL CH 4 /kgVS, whereas the wet digester was operated at an OLR of 0.9kgVS/m 3 day, generating about 320NL CH 4 /kgVS. The initial total volatile fatty acids concentrations for SADB and wet digestion were about 12,500mg/L and 4500mg/L, respectively. There were higher concentrations of ammonium and COD for the SADB compared to the wet one. The genomic analysis performed by high throughput sequencing returned a number of sequences for each sample ranging from 110,619 to 373,307. More than 93% were assigned to the Bacteria domain. Seven and nine major phyla were sequenced for the SADB and wet digestion, respectively, with Bacteroidetes, Firmicutes and Proteobacteria being the dominant phyla in both digesters. Taxonomic profiles suggested a methanogenic pathway characterized by a relevant syntrophic acetate-oxidizing metabolism mainly in the liquid digestate of the SADB. This result also confirms the benefits of liquid digestate recirculation for improving the efficiency of AD performed with high solids (>30%w/w) content. Copyright © 2016 Elsevier Ltd. All rights reserved.

Biogas and methane yield in response to co- and separate digestion of biomass wastes.

PubMed

Adelard, Laetitia; Poulsen, Tjalfe G; Rakotoniaina, Volana

2015-01-01

The impact of co-digestion as opposed to separate digestion, on biogas and methane yield (apparent synergetic effects) was investigated for three biomass materials (pig manure, cow manure and food waste) under mesophilic conditions over a 36 day period. In addition to the three biomass materials (digested separately), 13 biomass mixtures (co-digested) were used. Two approaches for modelling biogas and methane yield during co-digestion, based on volatile solids concentration and ultimate gas and methane potentials, were evaluated. The dependency of apparent synergetic effects on digestion time and biomass mixture composition was further assessed using measured cumulative biogas and methane yields and specific biogas and methane generation rates. Results indicated that it is possible, based on known volatile solids concentration and ultimate biogas or methane yields for a set of biomass materials digested separately, to accurately estimate gas yields for biomass mixtures made from these materials using calibrated models. For the biomass materials considered here, modelling indicated that the addition of pig manure is the main cause of synergetic effects. Co-digestion generally resulted in improved ultimate biogas and methane yields compared to separate digestion. Biogas and methane production was furthermore significantly higher early (0-7 days) and to some degree also late (above 20 days) in the digestion process during co-digestion. © The Author(s) 2014.

Predicting the apparent viscosity and yield stress of mixtures of primary, secondary and anaerobically digested sewage sludge: Simulating anaerobic digesters.

PubMed

Markis, Flora; Baudez, Jean-Christophe; Parthasarathy, Rajarathinam; Slatter, Paul; Eshtiaghi, Nicky

2016-09-01

Predicting the flow behaviour, most notably, the apparent viscosity and yield stress of sludge mixtures inside the anaerobic digester is essential because it helps optimize the mixing system in digesters. This paper investigates the rheology of sludge mixtures as a function of digested sludge volume fraction. Sludge mixtures exhibited non-Newtonian, shear thinning, yield stress behaviour. The apparent viscosity and yield stress of sludge mixtures prepared at the same total solids concentration was influenced by the interactions within the digested sludge and increased with the volume fraction of digested sludge - highlighted using shear compliance and shear modulus of sludge mixtures. However, when a thickened primary - secondary sludge mixture was mixed with dilute digested sludge, the apparent viscosity and yield stress decreased with increasing the volume fraction of digested sludge. This was caused by the dilution effect leading to a reduction in the hydrodynamic and non-hydrodynamic interactions when dilute digested sludge was added. Correlations were developed to predict the apparent viscosity and yield stress of the mixtures as a function of the digested sludge volume fraction and total solids concentration of the mixtures. The parameters of correlations can be estimated using pH of sludge. The shear and complex modulus were also modelled and they followed an exponential relationship with increasing digested sludge volume fraction. Copyright © 2016 Elsevier Ltd. All rights reserved.

78 FR 73551 - National Institute of Diabetes and Digestive and Kidney Diseases; Notice of Closed Meeting

Federal Register 2010, 2011, 2012, 2013, 2014

2013-12-06

... Diabetes and Digestive and Kidney Diseases; Notice of Closed Meeting Pursuant to section 10(d) of the... Diabetes and Digestive and Kidney Diseases Special Emphasis Panel; Digestive Diseases Ancillary Study. Date..., Digestive Diseases and Nutrition Research; 93.849, Kidney Diseases, Urology and Hematology Research...

75 FR 56119 - National Institute of Diabetes and Digestive and Kidney Diseases; Notice of Closed Meetings

Federal Register 2010, 2011, 2012, 2013, 2014

2010-09-15

... Diabetes and Digestive and Kidney Diseases; Notice of Closed Meetings Pursuant to section 10(d) of the... Diabetes and Digestive and Kidney Diseases Special Emphasis Panel, Fellowships in Digestive Diseases and... Institute of Diabetes and Digestive and Kidney Diseases Special Emphasis Panel. Kidney Disease Ancillary...

76 FR 3147 - National Institute of Diabetes and Digestive and Kidney Diseases; Notice of Closed Meetings

Federal Register 2010, 2011, 2012, 2013, 2014

2011-01-19

... Diabetes and Digestive and Kidney Diseases; Notice of Closed Meetings Pursuant to section 10(d) of the... Diabetes and Digestive and Kidney Diseases Special Emphasis Panel; Fellowships in Digestive Diseases and... Committee: National Institute of Diabetes and Digestive and Kidney Diseases Special Emphasis Panel...

78 FR 79706 - National Institute of Diabetes and Digestive and Kidney Diseases; Notice of Closed Meetings

Federal Register 2010, 2011, 2012, 2013, 2014

2013-12-31

... Diabetes and Digestive and Kidney Diseases; Notice of Closed Meetings Pursuant to section 10(d) of the... Diabetes and Digestive and Kidney Diseases Special Emphasis Panel; Fellowships in Digestive Diseases and... . Name of Committee: National Institute of Diabetes and Digestive and Kidney Diseases Special Emphasis...

78 FR 58322 - National Institute of Diabetes and Digestive and Kidney Diseases; Notice of Meeting

Federal Register 2010, 2011, 2012, 2013, 2014

2013-09-23

... Diabetes and Digestive and Kidney Diseases; Notice of Meeting Pursuant to section 10(d) of the Federal... Digestive and Kidney Diseases Initial Review Group, Digestive Diseases and Nutrition C Subcommittee. Date...; 93.848, Digestive Diseases and Nutrition Research; 93.849, Kidney Diseases, Urology and Hematology...

Evaluating the effects of activated carbon on methane generation and the fate of antibiotic resistant genes and class I integrons during anaerobic digestion of solid organic wastes.

PubMed

Zhang, Jingxin; Mao, Feijian; Loh, Kai-Chee; Gin, Karina Yew-Hoong; Dai, Yanjun; Tong, Yen Wah

2018-02-01

The effects of activated carbon (AC) on methane production and the fate of antibiotic resistance genes (ARGs) were evaluated through comparing the anaerobic digestion performance and transformation of ARGs among anaerobic mono-digestion of food waste, co-digestion of food waste and chicken manure, and co-digestion of food waste and waste activated sludge. Results showed that adding AC in anaerobic digesters improved methane yield by at least double through the enrichment of bacteria and archaea. Conventional digestion process showed ability in removing certain types of ARGs, such as tetA, tetX, sul1, sul2, cmlA, floR, and intl1. Supplementing AC in anaerobic digester enhanced the removal of most of the ARGs in mono-digestion of food waste. The effects tended to be minimal in co-digestion of co-substrates such as chicken manure and waste activated sludge, both of which contain a certain amount of antibiotics. Copyright © 2017 Elsevier Ltd. All rights reserved.

Digestion proteomics: tracking lactoferrin truncation and peptide release during simulated gastric digestion.

PubMed

Grosvenor, Anita J; Haigh, Brendan J; Dyer, Jolon M

2014-11-01

The extent to which nutritional and functional benefit is derived from proteins in food is related to its breakdown and digestion in the body after consumption. Further, detailed information about food protein truncation during digestion is critical to understanding and optimising the availability of bioactives, in controlling and limiting allergen release, and in minimising or monitoring the effects of processing and food preparation. However, tracking the complex array of products formed during the digestion of proteins is not easily accomplished using classical proteomics. We here present and develop a novel proteomic approach using isobaric labelling to mapping and tracking protein truncation and peptide release during simulated gastric digestion, using bovine lactoferrin as a model food protein. The relative abundance of related peptides was tracked throughout a digestion time course, and the effect of pasteurisation on peptide release assessed. The new approach to food digestion proteomics developed here therefore appears to be highly suitable not only for tracking the truncation and relative abundance of released peptides during gastric digestion, but also for determining the effects of protein modification on digestibility and potential bioavailability.

Biomethanation of Sewage Sludge with Food Waste Leachate Via Co-Digestion.

PubMed

Shin, Jingyeong; Kim, Young Beom; Jeon, Jong Hun; Choi, Sangki; Park, In Kyu; Kim, Young Mo

2017-08-28

Anaerobic mono- and co-digestion of sewage sludge and food waste leachate (FWL) were performed by assessing methane production and characterizing microbial communities. Anaerobic digestion (AD) of waste activated sludge (WAS) alone produced the lowest methane (281 ml CH 4 ), but an approximately 80% increase in methane production was achieved via co-digestion of WAS and FWL (506 ml CH 4 ). There were less differences in the diversity of bacterial communities in anaerobic digesters, while archaeal (ARC) and bacterial (BAC) amounts reflected AD performance. Compared with the total ARC and BAC amounts in the mono-digestion of WAS, the ARC and BAC amounts increased two and three times, respectively, during co-digestion of FWL and WAS. In characterized archaeal communities, the dominant ratio of hydrogenotrophic methanogens in the mono-digestion of WAS approached nearly a 1:1 ratio of the two acetoclastic and hydrogenotrophic methanogens in the co-digestion of FWL and WAS. The ARC/BAC ratio in the digesters varied in the range of 5.9% to 9.1%, indicating a positive correlation with the methane production of AD.

Immunohistochemical localization of hepatopancreatic phospholipase A2 in Hexaplex trunculus digestive cells.

PubMed

Zarai, Zied; Boulais, Nicholas; Karray, Aida; Misery, Laurent; Bezzine, Sofiane; Rebai, Tarek; Gargouri, Youssef; Mejdoub, Hafedh

2011-06-01

Mammalian sPLA2-IB localization cell are well characterized. In contrast, much less is known about aquatic primitive ones. The aquatic world contains a wide variety of living species and, hence represents a great potential for discovering new lipolytic enzymes and the mode of digestion of lipid food. The marine snail digestive phospholipase A2 (mSDPLA2) has been previously purified from snail hepatopancreas. The specific polyclonal antibodies were prepared and used for immunohistochimical and immunofluorescence analysis in order to determine the cellular location of mSDPLA2. Our results showed essentially that mSDPLA2 was detected inside in specific vesicles tentatively named (mSDPLA2+) granules of the digestive cells. No immunolabelling was observed in secretory zymogene-like cells. This immunocytolocalization indicates that lipid digestion in the snail might occur in specific granules inside the digestive cells. The cellular location of mSDPLA2 suggests that intracellular phospholipids digestion, like other food components digestion of snail diet, occurs in these digestive cells. The hepatopancreas of H. trunculus has been pointed out as the main region for digestion, absorption and storage of lipids.

Psychrophilic anaerobic digestion of guinea pig manure in low-cost tubular digesters at high altitude.

PubMed

Garfí, Marianna; Ferrer-Martí, Laia; Villegas, Vidal; Ferrer, Ivet

2011-05-01

Guinea pig is one of the most common livestock in rural communities of the Andes. The aim of this research was to study the anaerobic digestion of guinea pig manure in low-cost unheated tubular digesters at high altitude. To this end, the performance of two pilot digesters was monitored during 7 months; and two greenhouse designs were compared. In the dome roof digester the temperature and biogas production were significantly higher than in the shed roof digester. However, the biogas production rate was low (0.04 m(biogas)(3)m(digester)(-3) d(-1)), which is attributed to the low organic loading rate (0.6 kg(VS)m(digester)(-3)d(-1)) and temperature (23°C) of the system, among other factors. In a preliminary fertilization study, the potato yield per hectare was increased by 100% using the effluent as biofertilizer. Improving manure management techniques, increasing the organic loading rate and co digesting other substrates may be considered to enhance the process. Copyright © 2011 Elsevier Ltd. All rights reserved.

Kenaf Bast Fibers—Part I: Hermetical Alkali Digestion

DOE PAGES

Shi, Jinshu; Shi, Sheldon Q.; Barnes, H. Michael; ...

2011-01-01

The objective of this study was to develop a hermetical alkali digestion process to obtain single cellulosic fibers from kenaf bast. Kenaf bast were hermetically digested into single fiber using a 5% sodium hydroxide solution for one hour at four different temperatures (80 ° C, 110 ° C, 130 ° C, and 160 ° C). The hermetical digestion process used in this study produced fibers with high cellulose content (84.2–92.3%) due to the removal of lignin and hemicelluloses. The surface hardness and elastic modulus of the fibers digested at 130 ° C and 160 ° C were improved significantly comparedmore » with those digested at 80 ° C. The tensile modulus and tensile strength of the individual fibers reduced as the digestion temperature increased from 110 ° C to 160 ° C. Micropores were generated in fiber cell wall when the fibers were digested at 130 ° C and 160 ° C. The studies on the composites that were made from polypropylene reinforced with the digested fibers indicated that the compatibility between the digested fibers and polypropylene matrix was poor.« less

In vitro digestibility of goat milk and kefir with a new standardised static digestion method (INFOGEST cost action) and bioactivities of the resultant peptides.

PubMed

Nehir El, Sedef; Karakaya, Sibel; Simsek, Sebnem; Dupont, Didier; Menfaatli, Esra; Eker, Alper Tolga

2015-07-01

The hydrolysis degrees of goat milk and kefir during simulated gastrointestinal digestion and some bioactivities of the resulting peptides after fermentation and digestion were studied. A static in vitro digestion method by the COST FA1005 Action INFOGEST was used and goat milk and kefir were partially hydrolyzed during the gastric phase and had above 80% hydrolysis after duodenal digestion. There were no differences between the digestibility of goat milk and kefir (p > 0.05). Goat milk and kefir displayed about 7-fold antioxidant activity after digestion (p < 0.05). Fermentation showed no effect on the calcium-binding capacity of the samples (p > 0.05), however, after in vitro digestion calcium-binding capacity of the goat milk and kefir increased 2 and 5 fold, respectively (p < 0.05). Digested goat milk and kefir showed a higher dose-dependent inhibitory effect on α-amylase compared to undigested samples (p < 0.05). α-Glucosidase inhibitory activities and in vitro bile acid-binding capacities of the samples were not determined at the studied concentrations.

Unexpected High Digestion Rate of Cooked Starch by the Ct-Maltase-Glucoamylase Small Intestine Mucosal α-Glucosidase Subunit

PubMed Central

Lin, Amy Hui-Mei; Nichols, Buford L.; Quezada-Calvillo, Roberto; Avery, Stephen E.; Sim, Lyann; Rose, David R.; Naim, Hassan Y.; Hamaker, Bruce R.

2012-01-01

For starch digestion to glucose, two luminal α-amylases and four gut mucosal α-glucosidase subunits are employed. The aim of this research was to investigate, for the first time, direct digestion capability of individual mucosal α-glucosidases on cooked (gelatinized) starch. Gelatinized normal maize starch was digested with N- and C-terminal subunits of recombinant mammalian maltase-glucoamylase (MGAM) and sucrase-isomaltase (SI) of varying amounts and digestion periods. Without the aid of α-amylase, Ct-MGAM demonstrated an unexpected rapid and high digestion degree near 80%, while other subunits showed 20 to 30% digestion. These findings suggest that Ct-MGAM assists α-amylase in digesting starch molecules and potentially may compensate for developmental or pathological amylase deficiencies. PMID:22563462

Unexpected high digestion rate of cooked starch by the Ct-maltase-glucoamylase small intestine mucosal α-glucosidase subunit.

PubMed

Lin, Amy Hui-Mei; Nichols, Buford L; Quezada-Calvillo, Roberto; Avery, Stephen E; Sim, Lyann; Rose, David R; Naim, Hassan Y; Hamaker, Bruce R

2012-01-01

For starch digestion to glucose, two luminal α-amylases and four gut mucosal α-glucosidase subunits are employed. The aim of this research was to investigate, for the first time, direct digestion capability of individual mucosal α-glucosidases on cooked (gelatinized) starch. Gelatinized normal maize starch was digested with N- and C-terminal subunits of recombinant mammalian maltase-glucoamylase (MGAM) and sucrase-isomaltase (SI) of varying amounts and digestion periods. Without the aid of α-amylase, Ct-MGAM demonstrated an unexpected rapid and high digestion degree near 80%, while other subunits showed 20 to 30% digestion. These findings suggest that Ct-MGAM assists α-amylase in digesting starch molecules and potentially may compensate for developmental or pathological amylase deficiencies.

Effect of starvation and refeeding on digestive enzyme activities in sturgeon (Acipenser naccarii) and trout (Oncorhynchus mykiss).

PubMed

Furné, Miriam; García-Gallego, Manuel; Hidalgo, M Carmen; Morales, Amalia E; Domezain, Alberto; Domezain, Julio; Sanz, Ana

2008-04-01

The digestive enzyme activities were determined in Adriatic sturgeon and rainbow trout during starvation and refeeding period. Overall, the digestive enzyme activities are affected in the same sense in both species. The protease and lipase activities were decreased later than amylase activity. Even after 1 month of starvation, both species would be prepared to digest protein and lipids in an effective way. After 72 days of starvation, the digestive machinery of the sturgeon and of the trout shows an altered capacity to digest macronutrients. The capacity to digest proteins and lipids, after 60 days of refeeding, begins to become re-established in sturgeon and trout. In contrast, in this period, the capacity to digest carbohydrates remains depressed in both species.

Effects of varying dietary ratios of corn silage to alfalfa silage on digestion of neutral detergent fiber in lactating dairy cows.

PubMed

Lopes, F; Cook, D E; Combs, D K

2015-09-01

An in vivo study was performed to test an in vitro procedure and model that predicts total-tract neutral detergent fiber (NDF) digestibility for lactating dairy cattle. Corn silage (CS) and alfalfa silage (AS) were used as forages for this study. These forages had similar NDF composition, but fiber in the CS contained less indigestible NDF compared with AS (35.5 and 47.8% of indigestible NDF, respectively). The in vitro method estimated rate of digestion of alfalfa potentially digestible NDF to be approximately 2 times faster than CS fiber (6.11 and 3.21%/h, respectively). Four diets were formulated containing different proportions of CS to AS: 100CS:0AS, 67CS:33AS, 33CS:67AS, and 0CS:100AS, as percentage of diet DM basis. The objective was to construct diets that contained approximately similar levels of NDF but with different pool sizes and rates of digestion of potentially digestible NDF. Diets were fed to 8 ruminally cannulated, multiparous, lactating dairy cows in a replicated 4×4 Latin square with 21-d periods. Total-tract fiber digestibility and fiber digestion kinetic parameters observed in vivo were compared with the values predicted by the in vitro assay and model. Total-tract NDF digestibility coefficients were similar (41.8 and 40.6% of total NDF) for the in vitro and in vivo methods, respectively. As the proportion of dietary alfalfa increased, the digestibility of NDF increased. The rate of digestion of potentially digestible NDF predicted from the in vitro assay was also similar to what was observed in vivo. Results suggest that the in vitro total-tract NDF digestibility model could be used to predict rate of fiber digestion and NDF digestibility for lactating dairy cattle. Copyright © 2015 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Low cost digester monitoring under realistic conditions: Rural use of biogas and digestate quality.

PubMed

Castro, L; Escalante, H; Jaimes-Estévez, J; Díaz, L J; Vecino, K; Rojas, G; Mantilla, L

2017-09-01

The purpose of this work was to assess the behaviour of anaerobic digestion of cattle manure in a rural digester under realistic conditions, and estimate the quality and properties of the digestate. The data obtained during monitoring indicated that the digester operation was stable without risk of inhibition. It produced an average of 0.85Nm 3 biogas/d at 65.6% methane, providing an energy savings of 76%. In addition, the digestate contained high nutrient concentrations, which is an important feature of fertilizers. However, this method requires post-treatment due to the presence of pathogens. Copyright © 2017 Elsevier Ltd. All rights reserved.

Mechanism and Effect of Temperature on Variations in Antibiotic Resistance Genes during Anaerobic Digestion of Dairy Manure.

PubMed

Sun, Wei; Qian, Xun; Gu, Jie; Wang, Xiao-Juan; Duan, Man-Li

2016-07-22

Animal manure comprises an important reservoir for antibiotic resistance genes (ARGs), but the variation in ARGs during anaerobic digestion at various temperatures and its underlying mechanism remain unclear. Thus, we performed anaerobic digestion using dairy manure at three temperature levels (moderate: 20 °C, mesophilic: 35 °C, and thermophilic: 55 °C), to analyze the dynamics of ARGs and bacterial communities by quantitative PCR and 16S rRNA gene sequencing. We found that 8/10 detected ARGs declined and 5/10 decreased more than 1.0 log during thermophilic digestion, whereas only four and five ARGs decreased during moderate and mesophilic digestion, respectively. The changes in ARGs and bacterial communities were similar under the moderate and mesophilic treatments, but distinct from those in the thermophilic system. Potential pathogens such as Bacteroidetes, Proteobacteria, and Corynebacterium were removed by thermophilic digestion but not by moderate and mesophilic digestion. The bacterial community succession was the dominant mechanism that influenced the variation in ARGs and integrons during anaerobic digestion. Thermophilic digestion decreased the amount of mesophilic bacteria (Bacteroidetes and Proteobacteria) carrying ARGs. Anaerobic digestion generally decreased the abundance of integrons by eliminating the aerobic hosts of integrons (Actinomycetales and Bacilli). Thermophilic anaerobic digestion is recommended for the treatment and reuse of animal manure.

The Effects of Biopolymer Encapsulation on Total Lipids and Cholesterol in Egg Yolk during in Vitro Human Digestion

PubMed Central

Hur, Sun-Jin; Kim, Young-Chan; Choi, Inwook; Lee, Si-Kyung

2013-01-01

The purpose of this study was to examine the effect of biopolymer encapsulation on the digestion of total lipids and cholesterol in egg yolk using an in vitro human digestion model. Egg yolks were encapsulated with 1% cellulose, pectin, or chitosan. The samples were then passed through an in vitro human digestion model that simulated the composition of mouth saliva, stomach acid, and the intestinal juice of the small intestine by using a dialysis tubing system. The change in digestion of total lipids was monitored by confocal fluorescence microscopy. The digestion rate of total lipids and cholesterol in all egg yolk samples dramatically increased after in vitro human digestion. The digestion rate of total lipids and cholesterol in egg yolks encapsulated with chitosan or pectin was reduced compared to the digestion rate of total lipids and cholesterol in other egg yolk samples. Egg yolks encapsulated with pectin or chitosan had lower free fatty acid content, and lipid oxidation values than samples without biopolymer encapsulation. Moreover, the lipase activity decreased, after in vitro digestion, in egg yolks encapsulated with biopolymers. These results improve our understanding of the effects of digestion on total lipids and cholesterol in egg yolk within the gastrointestinal tract. PMID:23965957

Mechanism and Effect of Temperature on Variations in Antibiotic Resistance Genes during Anaerobic Digestion of Dairy Manure

NASA Astrophysics Data System (ADS)

Sun, Wei; Qian, Xun; Gu, Jie; Wang, Xiao-Juan; Duan, Man-Li

2016-07-01

Animal manure comprises an important reservoir for antibiotic resistance genes (ARGs), but the variation in ARGs during anaerobic digestion at various temperatures and its underlying mechanism remain unclear. Thus, we performed anaerobic digestion using dairy manure at three temperature levels (moderate: 20 °C, mesophilic: 35 °C, and thermophilic: 55 °C), to analyze the dynamics of ARGs and bacterial communities by quantitative PCR and 16S rRNA gene sequencing. We found that 8/10 detected ARGs declined and 5/10 decreased more than 1.0 log during thermophilic digestion, whereas only four and five ARGs decreased during moderate and mesophilic digestion, respectively. The changes in ARGs and bacterial communities were similar under the moderate and mesophilic treatments, but distinct from those in the thermophilic system. Potential pathogens such as Bacteroidetes, Proteobacteria, and Corynebacterium were removed by thermophilic digestion but not by moderate and mesophilic digestion. The bacterial community succession was the dominant mechanism that influenced the variation in ARGs and integrons during anaerobic digestion. Thermophilic digestion decreased the amount of mesophilic bacteria (Bacteroidetes and Proteobacteria) carrying ARGs. Anaerobic digestion generally decreased the abundance of integrons by eliminating the aerobic hosts of integrons (Actinomycetales and Bacilli). Thermophilic anaerobic digestion is recommended for the treatment and reuse of animal manure.

In vitro digestion testing of lipid-based delivery systems: calcium ions combine with fatty acids liberated from triglyceride rich lipid solutions to form soaps and reduce the solubilization capacity of colloidal digestion products.

PubMed

Devraj, Ravi; Williams, Hywel D; Warren, Dallas B; Mullertz, Anette; Porter, Christopher J H; Pouton, Colin W

2013-01-30

In vitro digestion testing is of practical importance to predict the fate of drugs administered in lipid-based delivery systems. Calcium ions are often added to digestion media to increase the extent of digestion of long-chain triglycerides (LCTs), but the effects they have on phase behaviour of the products of digestion, and consequent drug solubilization, are not well understood. This study investigates the effect of calcium and bile salt concentrations on the rate and extent of in vitro digestion of soybean oil, as well as the solubilizing capacity of the digestion products for two poorly water-soluble drugs, fenofibrate and danazol. In the presence of higher concentrations of calcium ions, the solubilization capacities of the digests were reduced for both drugs. This effect is attributed to the formation of insoluble calcium soaps, visible as precipitates during the digestions. This reduces the availability of liberated fatty acids to form mixed micelles and vesicles, thereby reducing drug solubilization. The use of high calcium concentrations does indeed force in vitro digestion of LCTs but may overestimate the extent of drug precipitation that occurs within the intestinal lumen. Copyright © 2012 Elsevier B.V. All rights reserved.

Gallbladder and Biliary Tract

MedlinePlus

... switch to the Professional version Home Digestive Disorders Biology of the Digestive System Gallbladder and Biliary Tract ... Version. DOCTORS: Click here for the Professional Version Biology of the Digestive System Overview of the Digestive ...

Redundancy in Anaerobic Digestion Microbiomes during Disturbances by the Antibiotic Monensin

PubMed Central

Spirito, Catherine M.; Daly, Sarah E.; Werner, Jeffrey J.

2018-01-01

ABSTRACT The antibiotic monensin is fed to dairy cows to increase milk production efficiency. A fraction of this monensin is excreted into the cow manure. Previous studies have found that cow manure containing monensin can negatively impact the performance of anaerobic digesters, especially upon first introduction. Few studies have examined whether the anaerobic digester microbiome can adapt to monensin during the operating time. Here, we conducted a long-term time series study of four lab-scale anaerobic digesters fed with cow manure. We examined changes in both the microbiome composition and function of the anaerobic digesters when subjected to the dairy antibiotic monensin. In our digesters, monensin was not rapidly degraded under anaerobic conditions. The two anaerobic digesters that were subjected to manure from monensin feed-dosed cows exhibited relatively small changes in microbiome composition and function due to relatively low monensin concentrations. At higher concentrations of monensin, which we dosed directly to control manure (from dairy cows without monensin), we observed major changes in the microbiome composition and function of two anaerobic digesters. A rapid introduction of monensin to one of these anaerobic digesters led to the impairment of methane production. Conversely, more gradual additions of the same concentrations of monensin to the other anaerobic digester led to the adaptation of the anaerobic digester microbiomes to the relatively high monensin concentrations. A member of the candidate OP11 (Microgenomates) phylum arose in this anaerobic digester and appeared to be redundant with certain Bacteroidetes phylum members, which previously were dominating. IMPORTANCE Monensin is a common antibiotic given to dairy cows in the United States and is partly excreted with dairy manure. An improved understanding of how monensin affects the anaerobic digester microbiome composition and function is important to prevent process failure for farm-based anaerobic digesters. This time series study demonstrates how anaerobic digester microbiomes are inert to low monensin concentrations and can adapt to relatively high monensin concentrations by redundancy in an already existing population. Therefore, our work provides further insight into the importance of microbiome redundancy in maintaining the stability of anaerobic digesters. PMID:29500266

Biokinetics and bacterial communities of propionate oxidizing bacteria in phased anaerobic sludge digestion systems.

PubMed

Zamanzadeh, Mirzaman; Parker, Wayne J; Verastegui, Yris; Neufeld, Josh D

2013-03-15

Phased anaerobic digestion is a promising technology and may be a potential source of bio-energy production. Anaerobic digesters are widely used for sewage sludge stabilization and thus a better understanding of the microbial process and kinetics may allow increased volatile solids reduction and methane production through robust process operation. In this study, we analyzed the impact of phase separation and operational conditions on the bio-kinetic characteristics and communities of bacteria associated with four phased anaerobic digestion systems. In addition to significant differences between bacterial communities associated with different digester operating temperatures, our results also revealed that bacterial communities in the phased anaerobic digestion systems differed between the 1st and 2nd phase digesters and we identified strong community composition correlations with several measured physicochemical parameters. The maximum specific growth rates of propionate oxidizing bacteria (POB) in the mesophilic and thermophilic 1st phases were 11 and 23.7 mgCOD mgCOD(-1) d(-1), respectively, while those of the mesophilic and thermophilic 2nd-phase digesters were 6.7 and 18.6 mgCOD mgCOD(-1) d(-1), respectively. Hence, the biokinetic characteristics of the POB population were dependent on the digester loading. In addition, we observed that the temperature dependency factor (θ) values were higher for the less heavily loaded digesters as compared to the values obtained for the 1st-phase digesters. Our results suggested the appropriate application of two sets of POB bio-kinetic that reflect the differing growth responses as a function of propionate concentration (and/or organic loading rates). Also, modeling acetogenesis in phased anaerobic sludge digestion systems will be improved considering a population shift in separate phases. On the basis of the bio-kinetic values estimated in various digesters, high levels of propionate in the thermophilic digesters may be best explained by the establishment of POB with low affinities (high K(s)) for propionate. Achieving low levels of propionate with either thermophilic or short HRT digesters is challenging and a relatively long HRT mesophilic digester should be employed for this purpose. Copyright © 2012 Elsevier Ltd. All rights reserved.

Anaerobic Digestion I. Sludge Treatment and Disposal Course #166. Instructor's Guide [and] Student Workbook.

ERIC Educational Resources Information Center

Arasmith, E. E.

This lesson is the first of a two-part series on anaerobic digestion. Topics discussed include the five basic functions of an anaerobic digester, basic theory of the biological processes involved, basic equipment necessary for digestion, and the products of digestion. The lesson includes an instructor's guide and student workbook. The instructor's…

Composition and Digestibility of Deer Browse in Southern Forests

Treesearch

Henry L. Short; Robert M. Blair; E.A. Epps

1975-01-01

Twigs were most nutritious and digestible during early growth in spring; they were high in fiber content and low in digestibility during summer, autumn, and winter. Evergreen leaves did not vary substantially in nutrient content and digestibility throughout the year. By contrast, leaves of deciduous species were reduced in quality and digestibility after leaf-fall....

Mesophilic and thermophilic anaerobic co-digestion of abattoir wastewater and fruit and vegetable waste in anaerobic sequencing batch reactors.

PubMed

Bouallagui, Hassib; Rachdi, Boutheina; Gannoun, Hana; Hamdi, Moktar

2009-06-01

Anaerobic co-digestion of fruit and vegetable waste (FVW) and abattoir wastewater (AW) was investigated using anaerobic sequencing batch reactors (ASBRs). The effects of hydraulic retention time (HRT) and temperature variations on digesters performances were examined. At both 20 and 10 days biogas production for co-digestion was greater thanks to the improved balance of nutrients. The high specific gas productions for the different digestion processes were 0.56, 0.61 and 0.85 l g(-1) total volatile solids (TVS) removal for digesters treating AW, FVW and AW + FVW, respectively. At an HRT of 20 days, biogas production rates from thermophilic digesters were higher on average than from mesophilic AW, FVW and AW + FVW digestion by 28.5, 44.5 and 25%, respectively. However, at 10 days of HRT results showed a decrease of biogas production rate for AW and AW + FVW digestion processes due to the high amount of free ammonia at high organic loading rate (OLR).

The interplay of α-amylase and amyloglucosidase activities on the digestion of starch in in vitro enzymic systems.

PubMed

Warren, Frederick J; Zhang, Bin; Waltzer, Gina; Gidley, Michael J; Dhital, Sushil

2015-03-06

In vitro hydrolysis assays are a key tool in understanding differences in rate and extent of digestion of starchy foods. They offer a greater degree of simplicity and flexibility than dynamic in vitro models or in vivo experiments for quantifiable, mechanistic exploration of starch digestion. In the present work the influence of α-amylase and amyloglucosidase activities on the digestion of maize and potato starch granules was measured using both glucose and reducing sugar assays. Data were analysed through initial rates of digestion, and by 1st order kinetics, utilising logarithm of slope (LOS) plots. The rate and extent of starch digestion was dependent on the activities of both enzymes and the type of starch used. Potato required more enzyme than maize to achieve logarithmic reaction curves, and complete digestion. The results allow targeted design of starch digestion experiments through a thorough understanding of the contributions of α-amylase and amyloglucosidase to digestion rates. Copyright © 2014 Elsevier Ltd. All rights reserved.

Assessment of in Vitro Digestibility of Dietary Carbohydrates Using Rat Small Intestinal Extract.

PubMed

Ferreira-Lazarte, Alvaro; Olano, Agustín; Villamiel, Mar; Moreno, F Javier

2017-09-13

There are few studies on the assessment of digestibility of nondigestible carbohydrates, despite their increasingly important role in human health. In vitro digestibility of a range of dietary carbohydrates classified as digestible (maltose, sucrose, and lactose), well-recognized (lactulose, fructooligosaccharides (FOS), and two types of galactooligosaccharides (GOS) differing in the predominant glycosidic linkage), and potential (lactosucrose and GOS from lactulose, OsLu) prebiotics using a rat small intestinal extract (RSIE) under physiological conditions of temperature and pH is described. Recognized and potential prebiotics were highly resistant to RSIE digestion although partial hydrolysis at different extents was observed. FOS and lactulose were the most resistant to digestion, followed closely by OsLu and more distantly by both types of GOS and lactosucrose. In GOS, β(1 → 6) linkages were more resistant to digestion than β(1 → 4) bonds. The reported in vitro digestion model is a useful, simple, and cost-effective tool to evaluate the digestibility of dietary oligosaccharides.

Recovery of failed solid-state anaerobic digesters.

PubMed

Yang, Liangcheng; Ge, Xumeng; Li, Yebo

2016-08-01

This study examined the performance of three methods for recovering failed solid-state anaerobic digesters. The 9-L digesters, which were fed with corn stover, failed at a feedstock/inoculum (F/I) ratio of 10 with negligible methane yields. To recover the systems, inoculum was added to bring the F/I ratio to 4. Inoculum was either added to the top of a failed digester, injected into it, or well-mixed with the existing feedstock. Digesters using top-addition and injection methods quickly resumed and achieved peak yields in 10days, while digesters using well-mixed method recovered slowly but showed 50% higher peak yields. Overall, these methods recovered 30-40% methane from failed digesters. The well-mixed method showed the highest methane yield, followed by the injection and top-addition methods. Recovered digesters outperformed digesters had a constant F/I ratio of 4. Slow mass transfer and slow growth of microbes were believed to be the major limiting factors for recovery. Copyright © 2016 Elsevier Ltd. All rights reserved.

Disinhibition of excessive volatile fatty acids to improve the efficiency of autothermal thermophilic aerobic sludge digestion by chemical approach.

PubMed

Jin, Ningben; Jin, Bo; Zhu, Nanwen; Yuan, Haiping; Ruan, Jianbo

2015-01-01

In this study, we explored a chemical approach to eliminate inhibition of excessive volatile fatty acids (VFAs) in autothermal thermophilic aerobic digestion (ATAD). Ferric nitrate, ferric chloride, potassium nitrate and potassium chloride were employed to demonstrate the combined action of ferric ion and nitrate ion. Supplementation of ferric nitrate in the sludge digestion system resulted in reducing the concentration of Total VFAs (TVFA) by round 5000mg/L and more than 2000mg/L of acetic acid at the end of digestion. Lower TVFA concentration contributed to faster sludge stabilization rate and the VS removal of ferric nitrate dosed digester achieved 38.18% after 12days digestion which was 9days in advance compared with the stabilization time of sludge in digester without chemicals addition. Lower concentrations of NH4(+)-N and SCOD in supernatant while higher content of TP in digestion sludge were obtained in digester with ferric nitrate added. Copyright © 2014 Elsevier Ltd. All rights reserved.

Enhanced methane generation during theromophilic co-digestion of confectionary waste and grease-trap fats and oils with municipal wastewater sludge.

PubMed

Gough, Heidi L; Nelsen, Diane; Muller, Christopher; Ferguson, John

2013-02-01

Recent interest in carbon-neutral biofuels has revived interest in co-digestion for methane generation. At wastewater treatment facilities, organic wastes may be co-digested with sludge using established anaerobic digesters. However, changes to organic loadings may induce digester instability, particularly for thermophilic digesters. To examine this problem, thermophilic (55 degrees C) co-digestion was studied for two food-industry wastes in semi-continuous laboratory digesters; in addition, the wastes' biochemical methane potentials were tested. Wastes with high chemical oxygen demand (COD) content were selected as feedstocks allowing increased input of potential energy to reactors without substantially altering volumetric loadings. Methane generation increased while reactor pH and volatile solids remained stable. Lag periods observed prior to methane stimulation suggested that acclimation of the microbial community may be critical to performance during co-digestion. Chemical oxygen demand mass balances in the experimental and control reactors indicated that all of the food industry waste COD was converted to methane.

Digestibility of Glyoxal-Glycated β-Casein and β-Lactoglobulin and Distribution of Peptide-Bound Advanced Glycation End Products in Gastrointestinal Digests.

PubMed

Zhao, Di; Li, Lin; Le, Thao T; Larsen, Lotte Bach; Su, Guoying; Liang, Yi; Li, Bing

2017-07-19

This work reports the influence of glyoxal (GO)-derived glycation on the gastrointestinal enzymatic hydrolysis of β-lactoglobulin and β-casein. Reduced digestibility of glycated proteins was found in both gastric and intestinal stage. Distribution of Maillard reaction products in digests with different molecular weight ranges was investigated subsequently. The colorless and brown MRPs largely presented in the digests smaller than 20 kDa. However, the resistance of fluorescent advanced glycation end products (AGEs) to enzymatic hydrolysis gradually increased during glycation, rendering fluorescent AGEs largely present in the digests larger than 20 kDa. No free N (ε)-carboxymethyllysine (CML) was detected in digests. The relative amount of CML in digests larger than 1 kDa was higher than that of Lys, demonstrating the hindrance of CML to enzymatic hydrolysis. This study highlights the resistance of GO-derived AGEs to digestive proteases via blockage of tryptic cleavage sites or steric hindrance, which is a barrier to the absorption of dietary AGEs.

Comparison of mesophilic and thermophilic anaerobic digestion of sugar beet pulp: performance, dewaterability and foam control.

PubMed

Suhartini, Sri; Heaven, Sonia; Banks, Charles J

2014-01-01

Digestion of sugar beet pulp was assessed in relation to biogas and methane production, foaming potential, and digestate dewaterability. Four 4-litre working volume digesters were operated mesophilically (37±0.5 °C) and four thermophilically (55±0.5 °C) over three hydraulic retention times. Digesters were operated in duplicate at organic loading rates (OLR) of 4 and 5 g volatile solids l(-1) day(-1) without water addition. Thermophilic digestion gave higher biogas and methane productivity than mesophilic and was able to operate at the higher OLR, where mesophilic digestion showed signs of instability. Digestate dewaterability was assessed using capillary suction time and frozen image centrifugation. The occurrence of, or potential for, stable foam formation was assessed using a foaming potential test. Thermophilic operation allowed higher loadings to be applied without loss of performance, and gave a digestate with superior dewatering characteristics and very little foaming potential. Copyright © 2013 Elsevier Ltd. All rights reserved.

Anaerobic co-digestion of dairy manure and potato waste

NASA Astrophysics Data System (ADS)

Yadanaparthi, Sai Krishna Reddy

Dairy and potato are two important agricultural commodities in Idaho. Both the dairy and potato processing industries produce a huge amount of waste which could cause environmental pollution. To minimize the impact of potential pollution associated with dairy manure (DM) and potato waste (PW), anaerobic co-digestion has been considered as one of the best treatment process. The purpose of this research is to evaluate the anaerobic co-digestion of dairy manure and potato waste in terms of process stability, biogas generation, construction and operating costs, and potential revenue. For this purpose, I conducted 1) a literature review, 2) a lab study on anaerobic co-digestion of dairy manure and potato waste at three different temperature ranges (ambient (20-25°C), mesophilic (35-37°C) and thermophilic (55-57°C) with five mixing ratios (DM:PW-100:0, 90:10, 80:20, 60:40, 40:60), and 3) a financial analysis for anaerobic digesters based on assumed different capital costs and the results from the lab co-digestion study. The literature review indicates that several types of organic waste were co-digested with DM. Dairy manure is a suitable base matter for the co-digestion process in terms of digestion process stability and methane (CH4) production (Chapter 2). The lab tests showed that co-digestion of DM with PW was better than digestion of DM alone in terms of biogas and CH4 productions (Chapter 3). The financial analysis reveals DM and PW can be used as substrate for full size anaerobic digesters to generate positive cash flow within a ten year time period. Based on this research, the following conclusions and recommendations were made: ▸ The ratio of DM:PW-80:20 is recommended at thermophilic temperatures and the ratio of DM:PW-90:10 was recommended at mesophilic temperatures for optimum biogas and CH4 productions. ▸ In cases of anaerobic digesters operated with electricity generation equipment (generators), low cost plug flow digesters (capital cost of 600/cow) operating at thermophilic temperatures are recommended. • The ratio of DM:PW-90:10 or 80:20 is recommended while operating low cost plug flow digesters at thermophilic temperatures. ▸ In cases of anaerobic digesters operated without electricity generation equipment (generators), completely mixed or high or low cost plug flow digesters can be used. • The ratio of DM:PW-80:20 is recommended for completely mixed digesters operated at thermophilic temperatures; • The ratio of DM:PW-90:10 or 80:20 is recommended for high cost plug flow digesters (capital cost of 1,000/cow) operated at thermophilic temperatures; • All of the four co-digested mixing ratios (i.e. DM:PW-90:10 or 80:20 or 60:40 or 40:60) are good for low cost plug flow digesters (capital cost of $600/cow) operated at thermophilic temperatures. The ratio of DM:PW-90:10 is recommended for positive cash flow within the ten year period if the low cost plug flow digesters are operated at mesophilic temperatures.

[Agreement between animal and vegetable protein digestibility measured in vivo and in vitro and its effect on the chemical score].

PubMed

Carias, D; Cioccia, A M; Hevia, P

1995-06-01

Protein digestibility is a key factor in the determination of protein quality using the chemical score. Since there are several methods available for determining protein digestibility the purpose of this study was to compare three methods in vitro (pH drop, pH stat and pepsin digestibility) and two methods in vivo (true and apparent digestibility in rats) in the determination of the protein digestibility of: casein, soy protein isolate, fish meal, black beans, corn meal and wheat flour. The results showed that in the case of highly digestible proteins all methods agreed very well. However, this agreement was much less apparent in the case of protein with digestibilities below 85%. As a result, the chemical score of these proteins varied substantially depending upon the method used to determine its digestibility. Thus, when the chemical score of the proteins analyzed was corrected by the true protein digestibility measured in rats, they ranked as: casein 83.56, soy 76.11, corn-beans mixtures (1:1) 58.14, fish meal 55.25, black beans 47.93, corn meal 46.06 and wheat flour 32.77. In contrast, when the chemical score of these proteins was corrected by the pepsin digestibility method, the lowest quality was assigned to fish meal. In summary, this results pointed out that for non conventional proteins of for known proteins which have been subjected to processing, protein digestibility should be measured in vivo.

Digestive Physiology of Octopus maya and O. mimus: Temporality of Digestion and Assimilation Processes

PubMed Central

Gallardo, Pedro; Olivares, Alberto; Martínez-Yáñez, Rosario; Caamal-Monsreal, Claudia; Domingues, Pedro M.; Mascaró, Maite; Sánchez, Ariadna; Pascual, Cristina; Rosas, Carlos

2017-01-01

Digestive physiology is one of the bottlenecks of octopus aquaculture. Although, there are successful experimentally formulated feeds, knowledge of the digestive physiology of cephalopods is fragmented, and focused mainly on Octopus vulgaris. Considering that the digestive physiology could vary in tropical and sub-tropical species through temperature modulations of the digestive dynamics and nutritional requirements of different organisms, the present review was focused on the digestive physiology timing of Octopus maya and Octopus mimus, two promising aquaculture species living in tropical (22–30°C) and sub-tropical (15–24°C) ecosystems, respectively. We provide a detailed description of how soluble and complex nutrients are digested, absorbed, and assimilated in these species, describing the digestive process and providing insight into how the environment can modulate the digestion and final use of nutrients for these and presumably other octopus species. To date, research on these octopus species has demonstrated that soluble protein and other nutrients flow through the digestive tract to the digestive gland in a similar manner in both species. However, differences in the use of nutrients were noted: in O. mimus, lipids were mobilized faster than protein, while in O. maya, the inverse process was observed, suggesting that lipid mobilization in species that live in relatively colder environments occurs differently to those in tropical ecosystems. Those differences are related to the particular adaptations of animals to their habitat, and indicate that this knowledge is important when formulating feed for octopus species. PMID:28620313

Investigating the fate of activated sludge extracellular proteins in sludge digestion using sodium dodecyl sulfate polyacrylamide gel electrophoresis.

PubMed

Park, Chul; Helm, Richard F; Novak, John T

2008-12-01

The fate of activated sludge extracellular proteins in sludge digestion was investigated using three different cation-associated extraction methods and sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Extraction methods used were the cation exchange resin (CER) method for extracting calcium (Ca2+) and magnesium (Mg2+), sulfide extraction for removing iron, and base treatment (pH 10.5) for dissolving aluminum. Extracellular polymeric substances extracted were then subjected to SDS-PAGE, and the resultant protein profiles were examined before and after sludge digestion. The SDS-PAGE results showed that three methods led to different SDS-PAGE profiles for both undigested and digested sludges. The results further revealed that CER-extracted proteins remained mainly undegraded in anaerobic digestion, but were degraded in aerobic digestion. While the fate of sulfide- and base-extracted proteins was not clear for aerobic digestion, their changes in anaerobic digestion were elucidated. Most sulfide-extracted proteins were removed by anaerobic digestion, while the increase in protein band intensity and diversity was observed for base-extracted proteins. These results suggest that activated sludge flocs contain different fractions of proteins that are distinguishable by their association with certain cations and that each fraction undergoes different fates in anaerobic and aerobic digestion. The proteins that were resistant to degradation and generated during anaerobic digestion were identified by liquid chromatography tandem mass spectrometry. Protein identification results and their putative roles in activated sludge and anaerobic digestion are discussed in this study.

Historical Analysis of Portuguese Primary School Textbooks (1920-2005) on the Topic of Digestion

ERIC Educational Resources Information Center

Carvalho, Graca S.; Silva, Rui; Clement, Pierre

2007-01-01

Our previous studies have shown that Portuguese primary school pupils and teachers have three main difficulties in the representation of the digestion process: the sequence of the digestive tract, blood absorption, and the relationship of the digestive function with other human functions. In this study we analysed the topic of digestion in 63…

Immunohistochemical localization of hepatopancreatic phospholipase A2 in Hexaplex Trunculus digestive cells

PubMed Central

2011-01-01

Background Mammalian sPLA2-IB localization cell are well characterized. In contrast, much less is known about aquatic primitive ones. The aquatic world contains a wide variety of living species and, hence represents a great potential for discovering new lipolytic enzymes and the mode of digestion of lipid food. Results The marine snail digestive phospholipase A2 (mSDPLA2) has been previously purified from snail hepatopancreas. The specific polyclonal antibodies were prepared and used for immunohistochimical and immunofluorescence analysis in order to determine the cellular location of mSDPLA2. Our results showed essentially that mSDPLA2 was detected inside in specific vesicles tentatively named (mSDPLA2+) granules of the digestive cells. No immunolabelling was observed in secretory zymogene-like cells. This immunocytolocalization indicates that lipid digestion in the snail might occur in specific granules inside the digestive cells. Conclusion The cellular location of mSDPLA2 suggests that intracellular phospholipids digestion, like other food components digestion of snail diet, occurs in these digestive cells. The hepatopancreas of H. trunculus has been pointed out as the main region for digestion, absorption and storage of lipids. PMID:21631952

Food Allergens: Is There a Correlation between Stability to Digestion and Allergenicity?

PubMed

Bøgh, Katrine Lindholm; Madsen, Charlotte Bernhard

2016-07-03

Food allergy is a major health problem in the Western countries, affecting 3-8% of the population. It has not yet been established what makes a dietary protein a food allergen. Several characteristics have been proposed to be shared by food allergens. One of these is resistance to digestion. This paper reviews data from digestibility studies on purified food allergens and evaluates the predictive value of digestibility tests on the allergenic potential. We point out that food allergens do not necessarily resist digestion. We discuss how the choice of in vitro digestibility assay condition and the method used for detection of residual intact protein as well as fragments hereof may greatly influence the outcome as well as the interpretation of results. The finding that digests from food allergens may retain allergenicity, stresses the importance of using immunological assays for evaluating the allergenic potential of food allergen digestion products. Studies assessing the allergenicity of digestion products, by either IgE-binding, elicitation or sensitizing capacity, shows that digestion may abolish, decrease, have no effect, or even increase the allergenicity of food allergens. Therefore, the predictive value of the pepsin resistance test for assessing the allergenic potential of novel proteins can be questioned.

Isolation, identification and utilization of thermophilic strains in aerobic digestion of sewage sludge.

PubMed

Liu, Shugen; Zhu, Nanwen; Li, Loretta Y; Yuan, Haiping

2011-11-15

Two representative thermophilic bacterial strains (T1 and T2) were isolated from a one-stage autothermal thermophilic aerobic digestion pilot-scale reactor. 16S rRNA gene analysis indicated that they were Hydrogenophilaceae and Xanthomonodaceae. These isolated strains were inoculated separately and/or jointly in sewage sludge, to investigate their effects on sludge stabilization under thermophilic aerobic digestion condition. Four digestion conditions were tested for 480 h. Digestion without inoculation and inoculation with strain T2, as well as joint- inoculation with strains T1 and T2, achieved 32.6%, 43.0%, and 38.2% volatile solids (VS) removal, respectively. Removal in a digester inoculated with stain T1 only reached 27.2%. For the first 144 h, the three inoculated digesters all experienced higher VS removal than the digester without inoculations. Both specific thermophilic strains and micro-environment significantly affected the VS removal. DGGE profiles revealed that the isolated strains T1 and T2 can successfully establish in the thermophilic digesters. Other viable bacteria (including anaerobic or facultative microbes) also appeared in the digestion system, enhancing the microbial activity. Copyright © 2011. Published by Elsevier Ltd.

Cow, sheep and llama manure at psychrophilic anaerobic co-digestion with low cost tubular digesters in cold climate and high altitude.

PubMed

Martí-Herrero, J; Alvarez, R; Cespedes, R; Rojas, M R; Conde, V; Aliaga, L; Balboa, M; Danov, S

2015-04-01

The aim of this research is to evaluate the co-digestion of cow and llama manure combined with sheep manure, in psychrophilic conditions and real field low cost tubular digesters adapted to cold climate. Four digesters were monitored in cold climate conditions; one fed with cow manure, a second one with llama manure, the third one with co-digestion of cow-sheep manure and the fourth one was fed with llama-sheep manure. The slurry had a mean temperature of 16.6 °C, the organic load rate was 0.44 kgvs m(-3) d(-1) and the hydraulic retention time was 80 days. After one hundred days biogas production was stable, as was the methane content and the pH of the effluent. The co-digestion of cow-sheep manure results in a biogas production increase of 100% compared to the mono-digestion of cow manure, while co-digestion of llama-sheep manure results in a decrease of 50% in biogas production with respect to mono-digestion of llama manure. Copyright © 2015 Elsevier Ltd. All rights reserved.

Blending protein separation and peptide analysis through real-time proteolytic digestion.

PubMed

Slysz, Gordon W; Schriemer, David C

2005-03-15

Typical liquid- or gel-based protein separations require enzymatic digestion as an important first step in generating protein identifications. Traditional protocols involve long-term proteolytic digestion of the separated protein, often leading to sample loss and reduced sensitivity. Previously, we presented a rapid method of proteolytic digestion that showed excellent digestion of resistant and low concentrations of protein without requiring reduction and alkylation. Here, we demonstrate on-line, real-time tryptic digestion in conjunction with reversed-phase protein separation. The studies were aimed at optimizing pH and ionic strength and the size of the digestion element, to produce maximal protein digestion with minimal effects on chromatographic integrity. Upon establishing optimal conditions, the digestion element was attached downstream from a capillary C4 reversed-phase column. A four-protein mixture was processed through the combined system, and the resulting peptides were analyzed on-line by electrospray mass spectrometry. Extracted ion chromatograms for protein chromatography based on peptide elution were generated. These were shown to emulate ion chromatograms produced in a subsequent run without the digestion element, based on protein elution. The methodology will enable rapid and sensitive analysis of liquid-based protein separations using the power of bottom-up proteomics methodologies.

Enhancement of anaerobic digestion efficiency of wastewater sludge and olive waste: Synergistic effect of co-digestion and ultrasonic/microwave sludge pre-treatment.

PubMed

Alagöz, B Aylin; Yenigün, Orhan; Erdinçler, Ayşen

2015-12-01

This study investigates the effect of ultrasonic and microwave pre-treatment on biogas production from the anaerobic co-digestion of olive pomace and wastewater sludges. It was found that co-digestion of wastewater sludge with olive pomace yielded around 0.21 L CH4/g VS added, whereas the maximum methane yields from the mono-digestion of olive pomace and un-pretreated wastewater sludges were 0.18 and 0.16L CH4/g VS added. In the same way, compared to mono-digestion of these substrates, co-digestion increased methane production by 17-31%. The microwave and ultrasonic pre-treatments applied to sludge samples prior to co-digestion process led to further increase in the methane production by 52% and 24%, respectively, compared to co-digestion with un-pretreated wastewater sludge. The highest biogas and methane yields were obtained from the co-digestion of 30 min microwave pre-treated wastewater sludges and olive pomace to be 0.46 L/g VS added and 0.32 L CH4/g VS added, respectively. Copyright © 2015 Elsevier Ltd. All rights reserved.

Effect of Different Heat Treatments on In Vitro Digestion of Egg White Proteins and Identification of Bioactive Peptides in Digested Products.

PubMed

Wang, Xuefen; Qiu, Ning; Liu, Yaping

2018-04-01

Chicken eggs are ingested by people after a series of processes, but to date, only a few studies have explored the nutrient variations caused by different heat treatments. In this work, the impacts of different heat treatments (4, 56, 65, and 100 °C on the in vitro digestibility of egg white proteins were investigated by hydrolysis with pepsin or pepsin + pancreatin to simulate human gastrointestinal digestion, and the digested products were identified using Nano-LC-ESI-MS/MS. Egg white proteins treated at 65 °C had the highest in vitro pepsin digestibility value whereas the pepsin + pancreatin digestibility increased significantly (P < 0.05) as the cooking temperature was raised. The molecular weight distribution of the digested products indicated that, when compared to pepsin-treated samples, pepsin + pancreatin-treated samples contained more low-molecular-weight peptides (m/z < 849.2) with higher signal intensities. The number of unique peptides identified in every digestion product showed a positive correlation with their in vitro digestibility. Additionally, bioactive peptides such as antioxidant, antimicrobial and antihypertensive peptides were found present in egg white digested products, especially in samples treated at 4 and 100 °C. These findings may facilitate a better understanding of nutritive values of egg white proteins and their digested products under different cooking temperatures, such as antibacterial and antioxidant peptides identified in the digestion samples treated, respectively at 4 and 100 °C. This study also provided information for improving the applications of eggs in the food industry as well as a theoretical basis for egg consumption. © 2018 Institute of Food Technologists®.

The role of two isoenzymes of alpha-amylase of Araucaria araucana (Araucariaceae) on the digestion of starch granules during germination.

PubMed

Waghorn, Juana J; del Pozo, Talía; Acevedo, Elba A; Cardemil, Liliana A

2003-03-01

Starch is the principal reserve of Araucaria araucana seeds, and it is hydrolysed during germination mainly by alpha-amylase. There are several alpha-amylase isoenzymes whose patterns change in the embryo and in the megagametophyte from the one observed in quiescent seeds (T(0)) to a different one observed 90 h after imbibition (T(90)). The objective of this research was to study the roles of two purified alpha-amylase isoenzymes by in vitro digestion of starch granules extracted from the tissues at two times of imbibition: one is abundant in quiescent seeds and the other is abundant after 90 h of imbibition. The isoenzymes digested the starch granules of their own stage of germination better, since the isoenzyme T(0) digested starch granules mainly from quiescent seeds, while the isoenzyme T(90) digested starch mainly at 90 h of imbibition. The sizes of the starch granule and the tissue from which these granules originated make a difference to digestion by the isoenzymes. Embryonic isoenzyme T(0) digested large embryonic starch granules better than small and medium-sized granules, and better than those isolated from megagametophytes. Similarly isoenzyme T(90) digested small embryonic starch granules better than medium-sized and large granules, and better than those isolated from megagametophytes. However, a mixture of partially purified megagametophytic isoenzymes T(0) and T(90) digested the megagametophytic granules better than those isolated from embryos. Studies of in vitro sequential digestion of starch granules with these isoenzymes corroborated their specificity. The isoenzyme T(90) digested starch granules previously digested by the isoenzyme T(0). This suggests that in vivo these two isoenzymes may act sequentially in starch granule digestion.

Anaerobic digestion of paunch in a CSTR for renewable energy production and nutrient mineralization

DOE Office of Scientific and Technical Information (OSTI.GOV)

Nkemka, Valentine Nkongndem; Marchbank, Douglas H.; Hao, Xiying, E-mail: xiying.hao@agr.gc.ca

Highlights: • Anaerobic digestion and nutrient mineralization of paunch in a CSTR. • Low CH{sub 4} yield and high CH{sub 4} productivity was obtained at an OLR of 2.8 g VS L{sup −1} day{sup −1.} • Post-digestion of the digestate resulted in a CH{sub 4} yield of 0.067 L g{sup −1} VS. • Post-digestion is recommended for further digestate stabilization. - Abstract: A laboratory study investigated the anaerobic digestion of paunch in a continuous stirred tank reactor (CSTR) for the recovery of biogas and mineralization of nutrients. At an organic loading rate (OLR) of 2.8 g VS L{sup −1} day{supmore » −1} with a 30-day hydraulic retention time (HRT), a CH{sub 4} yield of 0.213 L g{sup −1} VS and CH{sub 4} production rate of 0.600 L L{sup −1} day{sup −1} were obtained. Post-anaerobic digestion of the effluent from the CSTR for 30 days at 40 °C recovered 0.067 L g{sup −1} VS as CH{sub 4}, which was 21% of the batch CH{sub 4} potential. Post-digestion of the effluent from the digestate obtained at this OLR is needed to meet the stable effluent criteria. Furthermore, low levels of soluble ions such as K{sup +}, Ca{sup 2+} and Mg{sup 2+} were found in the liquid fraction of the digestate and the remainder could have been retained in the solid digestate fraction. This study demonstrates the potential of biogas production from paunch in providing renewable energy. In addition, recovery of plant nutrients in the digestate is important for a sustainable agricultural system.« less

[The digestive characteristics of fishes (the biocenotic aspects)].

PubMed

Kuz'mina, V V

1996-03-01

The cavity, membrane and intracellular digestion is added by two additional types of digestion in the fish: a symbiont digestion and an induced autolysis. As the intensity of processes maintained by the enzyme systems of the victim and a symbiont depends upon the structure and conditions of the biocenosis, the latter are regarded as an ecological component. The enzymes of microflora and objects of feeding contribute much to digestive processes in fish and take part in the adaptation of their digestive system to the character of the food and specifics of the biocenosis.

Up-to-date modification of the anaerobic sludge digestion process introducing a separate sludge digestion mode.

PubMed

Sato, K; Ochi, S; Mizuochi, M

2001-01-01

Sewage treatment plants in Japan are subjected to advanced treatment to remove nutrients and hence control eutrophication problems in lakes and bays. This paper discusses the advantages and disadvantages of the separate digestion treatment mode for sludge generated from advanced wastewater treatment. In the separate digestion only primary sludge is digested and the excess activated sludge is directly dewatered. Separate digestion can reduce the return load of nutrients to approximately one third, and has major potential for the beneficial use of sludge.

Digestibility marker and ileal amino acid digestibility in phytase-supplemented soybean or canola meals for growing pigs.

PubMed

Favero, A; Ragland, D; Vieira, S L; Owusu-Asiedu, A; Adeola, O

2014-12-01

Two experiments using soybean meal (SBM) or canola meal (CM) were conducted to investigate whether the choice of digestibility marker influenced the apparent ileal digestibility (AID) or standardized ileal digestibility (SID) of N and AA in diets supplemented with phytase. In each experiment, 18 barrows fitted with T-cannulas at the ileocecal junction were assigned to 3 diets consisting of a N-free diet to determine endogenous losses of N and AA, a semipurified diet (SBM in Exp. 1 or CM in Exp. 2), and the semipurified diet supplemented with phytase at 1,000 phytase units/kg. Three digestibility markers including acid-insoluble ash (AIA), chromic oxide (Cr2O3), and titanium dioxide (TiO2) were added to each diet at 3 g/kg. Each diet was fed for 7 d, consisting of a 5-d adjustment and a 2-d collection of ileal digesta. In both studies, basal ileal endogenous losses determined with Cr2O3 as a digestibility marker were lower (P<0.01) than with those determined with AIA or TiO2 digestibility markers. Using SBM as the protein source in Exp. 1, there was no interaction between phytase and digestibility marker on AID or SID of AA. The AID of N and AA in SBM using AIA as a digestibility marker tended to be lower (P<0.1) compared with Cr2O3 or TiO2 digestibility markers. Phytase supplementation increased (P<0.001) the AID of Ca and P. The use of AIA or Cr2O3 digestibility marker tended to be associated with lower (P<0.1) SID values compared with TiO2. Phytase did not affect the SID of N or any AA in SBM except for Met, for which there was an increase (P<0.05) with phytase supplementation. Using CM as the protein source in Exp. 2, there were significant interactions between digestibility marker and phytase. Phytase supplementation had effects (P<0.01) on AID or SID when Cr2O3 or TiO2 was used as the digestibility marker. With Cr2O3 or TiO2 as the digestibility marker in the CM diets, phytase supplementation increased (P<0.05) the SID of N and all AA (except Trp). There was no SID of N or AA response to phytase supplementation of CM when AIA was used as a digestibility marker. In contrast, there were no clear improvements in AA digestibility from phytase supplementation for SBM. Phytase effects on AID or SID of AA were dependent on the digestibility marker used in diets when CM was used as the protein source but not when SBM was used as the protein source. Therefore, AA digestibility response to phytase supplementation may depend on the protein being evaluated as well as the choice of digestibility marker.

Biochemical, transcriptomic and proteomic analyses of digestion in the scorpion Tityus serrulatus: insights into function and evolution of digestion in an ancient arthropod.

PubMed

Fuzita, Felipe J; Pinkse, Martijn W H; Patane, José S L; Juliano, Maria A; Verhaert, Peter D E M; Lopes, Adriana R

2015-01-01

Scorpions are among the oldest terrestrial arthropods and they have passed through small morphological changes during their evolutionary history on land. They are efficient predators capable of capturing and consuming large preys and due to envenomation these animals can become a human health challenge. Understanding the physiology of scorpions can not only lead to evolutionary insights but also is a crucial step in the development of control strategies. However, the digestive process in scorpions has been scarcely studied. In this work, we describe the combinatory use of next generation sequencing, proteomic analysis and biochemical assays in order to investigate the digestive process in the yellow scorpion Tityus serrulatus, mainly focusing in the initial protein digestion. The transcriptome generated database allowed the quantitative identification by mass spectrometry of different enzymes and proteins involved in digestion. All the results suggested that cysteine cathepsins play an important role in protein digestion. Two digestive cysteine cathepsins were isolated and characterized presenting acidic characteristics (pH optima and stability), zymogen conversion to the mature form after acidic activation and a cross-class inhibition by pepstatin. A more elucidative picture of the molecular mechanism of digestion in a scorpion was proposed based on our results from Tityus serrulatus. The midgut and midgut glands (MMG) are composed by secretory and digestive cells. In fasting animals, the secretory granules are ready for the next predation event, containing enzymes needed for alkaline extra-oral digestion which will compose the digestive fluid, such as trypsins, astacins and chitinase. The digestive vacuoles are filled with an acidic proteolytic cocktail to the intracellular digestion composed by cathepsins L, B, F, D and legumain. Other proteins as lipases, carbohydrases, ctenitoxins and a chitolectin with a perithrophin domain were also detected. Evolutionarily, a large gene duplication of cathepsin L occurred in Arachnida with the sequences from ticks being completely divergent from other arachnids probably due to the particular selective pressures over this group.

Performance evaluation of a high-pressure microwave-assisted flow digestion system for juice and milk sample preparation.

PubMed

Marques, Thiago L; Wiltsche, Helmar; Nóbrega, Joaquim A; Winkler, Monika; Knapp, Günter

2017-07-01

Acid digestion is usually required for metal determination in food samples. However, this step is usually performed in batch mode which is time consuming, labor intensive, and may lead to sample contamination. Flow digestion can overcome these limitations. In this work, the performance of a high-pressure microwave-assisted flow digestion system with a large volume reactor was evaluated for liquid samples high in sugar and fat (fruit juice and milk). The digestions were carried out in a coiled perfluoroalkoxy (PFA) tube reactor (13.5 mL) installed inside an autoclave pressurized with 40 bar nitrogen. The system was operated at 500 W microwave power and 5.0 mL min -1 carrier flow rate. Digestion conditions were optimized with phenylalanine, as this substance is known to be difficult to digest completely. The combinations of HCl or H 2 O 2 with HNO 3 increased the digestion efficiency of phenylalanine, and the residual carbon content (RCC) was around 50% when 6.0% V/V HCl or H 2 O 2 was used in combination with 32% V/V HNO 3 . Juice samples were digested with 3.7 mol L -1 HNO 3 and 0.3 mol L -1 HCl, and the RCC was 16 and 29% for apple and mango juices, respectively. Concentrated HNO 3 (10.5 mol L -1 ) was successfully applied for digesting milk samples, and the RCCs were 23 and 25% for partially skimmed and whole milk, respectively. Accuracy and precision of the flow digestion procedure were compared with reference digestions using batch mode closed vessel microwave-assisted digestion and no statistically significant differences were encountered at the 95% confidence level. Graphical abstract Application of a high-pressure microwave-assisted flow digestion system for fruit juice and milk sample preparation.

Predicting the apparent viscosity and yield stress of digested and secondary sludge mixtures.

PubMed

Eshtiaghi, Nicky; Markis, Flora; Zain, Dwen; Mai, Kiet Hung

2016-05-15

The legal banning of conventional sludge disposal methods such as landfill has led to a global movement towards achieving a sustainable sludge management strategy. Reusing sludge for energy production (biogas production) through the anaerobic digestion of sludge can provide a sustainable solution. However, for the optimum performance of digesters with minimal use of energy input, operating conditions must be regulated in accordance with the rheological characteristics of the sludge. If it is assumed that only secondary sludge enters the anaerobic digesters, an impact of variations to the solids concentration and volume fraction of each sludge type must be investigated to understand how the apparent viscosity and yield stress of the secondary and digested sludge mixture inside the digesters changes. In this study, five different total solids concentration of secondary and digested sludge were mixed at different digested sludge volume fractions ranging from 0 to 1. It was found that if secondary sludge was mixed with digested sludge at the same total solids concentration, the apparent viscosity and the yield stress of the mixture increased exponentially by increasing the volume fraction of digested sludge. However, if secondary sludge was added to digested sludge with a different solids concentration, the apparent viscosity and yield stress of the resulting mixed sludge was controlled by the concentrated sludge regardless of its type. Semi - empirical correlations were proposed to predict the apparent viscosity and yield stress of the mixed digested and secondary sludge. A master curve was also developed to predict the flow behaviour of sludge mixtures regardless of the total solid concentration and volume fraction of each sludge type within the studied solids concentration range of 1.4 and 7%TS. This model can be used for digesters optimization and design by predicting the rheology of sludge mixture inside digester. Copyright © 2016 Elsevier Ltd. All rights reserved.

Anaerobic co-digestion of recalcitrant agricultural wastes: Characterizing of biochemical parameters of digestate and its impacts on soil ecosystem.

PubMed

Muscolo, Adele; Settineri, Giovanna; Papalia, Teresa; Attinà, Emilio; Basile, Carmelo; Panuccio, Maria Rosaria

2017-05-15

Anaerobic digestion (AD) of organic wastes is a promising alternative to landfilling for reducing Greenhouse Gas Emission (GHG) and it is encouraged by current regulation in Europe. Biogas-AD produced, represents a useful source of green energy, while its by-product (digestate) is a waste, that needs to be safely disposal. The sustainability of anaerobic digestion plants partly depends on the management of their digestion residues. This study has been focused on the environmental and economic benefits of co-digest recalcitrant agricultural wastes such olive wastes and citrus pulp, in combination with livestock wastes, straw and cheese whey for biogas production. The aim of this work was to investigate the effects of two different bioenergy by-products on soil carbon stock, enzymes involved in nutrient cycling and microbial content. The two digestates were obtained from two plants differently fed: the first plant (Uliva) was powered with 60% of recalcitrant agricultural wastes, and 40% of livestock manure milk serum and maize silage. The second one (Fattoria) was fed with 40% of recalcitrant agricultural wastes and 60% of livestock manure, milk serum and maize silage. Each digestate, separated in liquid and solid fractions, was added to the soil at different concentrations. Our results evidenced that mixing and type of input feedstock affected the composition of digestates. Three months after treatments, our results showed that changes in soil chemical and biochemical characteristics depended on the source of digestate, the type of fraction and the concentration used. The mainly affected soil parameters were: Soil Organic Matter (SOM), Microbial Biomass Carbon (MBC), Fluorescein Diacetate Hydrolysis (FDA), Water Soluble Phenol (WSP) and Catalase (CAT) that can be used to assess the digestate agronomical feasibility. These results show that the agronomic quality of a digestate is strictly dependent on percentage and type of feedstocks that will be used to power the digester. Copyright © 2017 Elsevier B.V. All rights reserved.

Apolipoprotein B and E genetic polymorphisms in the Cayapa Indians of Ecuador.

PubMed

Scacchi, R; Corbo, R M; Rickards, O; Mantuano, E; Guevara, A; De Stefano, G F

1997-06-01

Three DNA polymorphisms (XbaI, EcoRI, and Ins/Del) of the apolipoprotein B (APOB) gene and the CfoI polymorphism of the APOE gene were investigated in a sample of 96 Cayapa Indians from Ecuador. The frequencies of the X+ (0.182), R+ (1.000), and Del alleles (0.432) at the three APOB sites were found to be higher than and to differ significantly from those reported for East Asians. No comparisons could be made between the Cayapa and other native Americans because of the lack of data on these sites. We observed in our sample that, like native American populations but unlike East Asians, the APOE allele frequencies were characterized by the absence of the APOE*2 allele and by a high frequency of the APOE*4 allele (0.280). Besides a probable drift effect, the high APOE*4 value was tentatively attributed to an effect of selection. Because this allele enhances the absorption of cholesterol by the intestine, it could confer an advantage to carriers in an unfavorable environment (i.e., diet poor in cholesterol).

Clonal structure in Ichthyobacterium seriolicida, the causative agent of bacterial haemolytic jaundice in yellowtail, Seriola quinqueradiata, inferred from molecular epidemiological analysis.

PubMed

Matsuyama, T; Fukuda, Y; Sakai, T; Tanimoto, N; Nakanishi, M; Nakamura, Y; Takano, T; Nakayasu, C

2017-08-01

Bacterial haemolytic jaundice caused by Ichthyobacterium seriolicida has been responsible for mortality in farmed yellowtail, Seriola quinqueradiata, in western Japan since the 1980s. In this study, polymorphic analysis of I. seriolicida was performed using three molecular methods: amplified fragment length polymorphism (AFLP) analysis, multilocus sequence typing (MLST) and multiple-locus variable-number tandem repeat analysis (MLVA). Twenty-eight isolates were analysed using AFLP, while 31 isolates were examined by MLST and MLVA. No polymorphisms were identified by AFLP analysis using EcoRI and MseI, or by MLST of internal fragments of eight housekeeping genes. However, MLVA revealed variation in repeat numbers of three elements, allowing separation of the isolates into 16 sequence types. The unweighted pair group method using arithmetic averages cluster analysis of the MLVA data identified four major clusters, and all isolates belonged to clonal complexes. It is likely that I. seriolicida populations share a common ancestor, which may be a recently introduced strain. © 2016 John Wiley & Sons Ltd.

Analysis of ELA-DQB exon 2 polymorphism in Argentine Creole horses by PCR-RFLP and PCR-SSCP.

PubMed

Villegas-Castagnasso, E E; Díaz, S; Giovambattista, G; Dulout, F N; Peral-García, P

2003-08-01

The second exon of equine leucocyte antigen (ELA)-DQB genes was amplified from genomic DNA of 32 Argentine Creole horses by PCR. Amplified DNA was analysed by PCR-restriction fragment length polymorphism (RFLP) and PCR-single-strand conformation polymorphism (SSCP). The PCR-RFLP analysis revealed two HaeIII patterns, four RsaI patterns, five MspI patterns and two HinfI patterns. EcoRI showed no variation in the analysed sample. Additional patterns that did not account for known exon 2 DNA sequences were observed, suggesting the existence of novel ELA-DQB alleles. PCR-SSCP analysis exhibited seven different band patterns, and the number of bands per animal ranged from four to nine. Both methods indicated that at least two DQB genes are present. The presence of more than two alleles in each animal showed that the primers employed in this work are not specific for a unique DQB locus. The improvement of this PCR-RFLP method should provide a simple and rapid technique for an accurate definition of ELA-DQB typing in horses.

Methods of analysis by the U.S. Geological Survey National Water Quality Laboratory; comparison of a nitric acid in-bottle digestion procedure to other whole-water digestion procedures

USGS Publications Warehouse

Garbarino, John R.; Hoffman, Gerald L.

1999-01-01

A hydrochloric acid in-bottle digestion procedure is used to partially digest wholewater samples prior to determining recoverable elements by various analytical methods. The use of hydrochloric acid is problematic for some methods of analysis because of spectral interference. The inbottle digestion procedure has been modified to eliminate such interference by using nitric acid instead of hydrochloric acid in the digestion. Implications of this modification are evaluated by comparing results for a series of synthetic whole-water samples. Results are also compared with those obtained by using U.S. Environmental Protection Agency (1994) (USEPA) Method 200.2 total-recoverable digestion procedure. Percentage yields that use the nitric acid inbottle digestion procedure are within 10 percent of the hydrochloric acid in-bottle yields for 25 of the 26 elements determined in two of the three synthetic whole-water samples tested. Differences in percentage yields for the third synthetic whole-water sample were greater than 10 percent for 16 of the 26 elements determined. The USEPA method was the most rigorous for solubilizing elements from particulate matter in all three synthetic whole-water samples. Nevertheless, the variability in the percentage yield by using the USEPA digestion procedure was generally greater than the in-bottle digestion procedure, presumably because of the difficulty in controlling the digestion conditions accurately.

Wood digestion in Pselactus spadix Herbst--a weevil attacking marine timber structures.

PubMed

Oevering, Pascal; Pitman, Andrew J; Pandey, Krishna K

2003-04-01

Pselactus spadix tunnels timber structures in the marine environment. Recent studies reported a cosmopolitan distribution for this weevil, which is frequently found in harbour and port areas. P. spadix feeds on timber (hardwood and softwood) in immature and adult life stages, but its digestion of wood components had not been investigated. Using dry weight analyses of tunnel walls and frass produced, P. spadix adults consumed Scots pine with soft rot decay at a rate of 1.59 +/- 0.37 mg d-1 and the digestibility of this substrate was 57.96 +/- 5.89 (i.e. for 100 mg consumed SR-pine, 58 mg was digested). Using gravimetric analysis to quantify structural wood components in tunnel walls and frass, P. spadix adults were found to digest cellulose, lignin and hemicellulose with digestibility coefficients of 82.2, 41.2 and 14.5 respectively. Fourier Transform Infrared (FTIR) spectroscopy analyses of tunnel walls and frass of adults and larvae from soft rotted pine also indicated digestion of all structural components, with larvae digesting cellulose and lignin more efficiently than adults. When FTIR was employed to analyse adult tunnel walls and frass from undecayed pine, cellulose and hemicellulose were digested, but no evidence of lignin digestion was found. This study shows that adults digest lignin when soft rot is present and suggests a symbiotic function of wood degrading microorganisms.

Anaerobic digestibility of beef hooves with swine manure or slaughterhouse sludge.

PubMed

Xia, Yun; Wang, Ding-Kang; Kong, Yunhong; Ungerfeld, Emilio M; Seviour, Robert; Massé, Daniel I

2015-04-01

Anaerobic digestion is an effective method for treating animal by-products, generating at the same time green energy as methane (CH4). However, the methods and mechanisms involved in anaerobic digestion of α-keratin wastes like hair, nails, horns and hooves are still not clear. In this study we investigated the feasibility of anaerobically co-digesting ground beef hooves in the presence of swine manure or slaughterhouse sludge at 25 °C using eight 42-L Plexiglas lab-scale digesters. Our results showed addition of beef hooves statistically significantly increased the rate of CH4 production with swine manure, but only increased it slightly with slaughterhouse sludge. After 90-day digestion, 73% of beef hoof material added to the swine manure-inoculated digesters had been converted into CH4, which was significantly higher than the 45% level achieved in the slaughterhouse sludge inoculated digesters. BODIPY-Fluorescent casein staining detected proteolytic bacteria in all digesters with and without added beef hooves, and their relative abundances corresponded to the rate of methanogenesis of the digesters with the different inocula. Fluorescence in situ hybridization in combination with BODIPY-Fluorescent casein staining identified most proteolytic bacteria as members of genus Alkaliphilus in the subfamily Clostridiaceae 2 of family Clostridiaceae. They thus appear to be the bacteria mainly responsible for digestion of beef hooves. Crown Copyright © 2014. Published by Elsevier Ltd. All rights reserved.

Mechanism and Effect of Temperature on Variations in Antibiotic Resistance Genes during Anaerobic Digestion of Dairy Manure

PubMed Central

Sun, Wei; Qian, Xun; Gu, Jie; Wang, Xiao-Juan; Duan, Man-Li

2016-01-01

Animal manure comprises an important reservoir for antibiotic resistance genes (ARGs), but the variation in ARGs during anaerobic digestion at various temperatures and its underlying mechanism remain unclear. Thus, we performed anaerobic digestion using dairy manure at three temperature levels (moderate: 20 °C, mesophilic: 35 °C, and thermophilic: 55 °C), to analyze the dynamics of ARGs and bacterial communities by quantitative PCR and 16S rRNA gene sequencing. We found that 8/10 detected ARGs declined and 5/10 decreased more than 1.0 log during thermophilic digestion, whereas only four and five ARGs decreased during moderate and mesophilic digestion, respectively. The changes in ARGs and bacterial communities were similar under the moderate and mesophilic treatments, but distinct from those in the thermophilic system. Potential pathogens such as Bacteroidetes, Proteobacteria, and Corynebacterium were removed by thermophilic digestion but not by moderate and mesophilic digestion. The bacterial community succession was the dominant mechanism that influenced the variation in ARGs and integrons during anaerobic digestion. Thermophilic digestion decreased the amount of mesophilic bacteria (Bacteroidetes and Proteobacteria) carrying ARGs. Anaerobic digestion generally decreased the abundance of integrons by eliminating the aerobic hosts of integrons (Actinomycetales and Bacilli). Thermophilic anaerobic digestion is recommended for the treatment and reuse of animal manure. PMID:27444518

Effect of short-time aerobic digestion on bioflocculation of extracellular polymeric substances from waste activated sludge.

PubMed

Zhang, Zhiqiang; Zhang, Jiao; Zhao, Jianfu; Xia, Siqing

2015-02-01

The effect of short-time aerobic digestion on bioflocculation of extracellular polymeric substances (EPSs) from waste activated sludge (WAS) was investigated. Bioflocculation of the EPS was found to be enhanced by 2∼6 h of WAS aerobic digestion under the conditions of natural sludge pH (about 7), high sludge concentration by gravity thickening, and dissolved oxygen of about 2 mg/L. With the same EPS extraction method, the total suspended solid content reduction of 0.20 and 0.36 g/L and the volatile suspended solid content reduction of 0.19 and 0.26 g/L were found for the WAS samples before and after aerobic digestion of 4 h. It indicates that more EPS is produced by short-time aerobic digestion of WAS. The scanning electron microscopy images of the WAS samples before and after aerobic digestion of 4 h showed that more EPS appeared on the surface of zoogloea by aerobic digestion, which reconfirmed that WAS aerobic digestion induced abundant formation of EPS. By WAS aerobic digestion, the flocculating rate of the EPS showed about 31 % growth, almost consistent with the growth of its yield (about 34 %). The EPSs obtained before and after the aerobic digestion presented nearly the same components, structures, and Fourier transform infrared spectra. These results revealed that short-time aerobic digestion of WAS enhanced the flocculation of the EPS by promoting its production.

High-solids anaerobic co-digestion of sewage sludge and food waste in comparison with mono digestions: stability and performance.

PubMed

Dai, Xiaohu; Duan, Nina; Dong, Bin; Dai, Lingling

2013-02-01

System stability and performance of high-solids anaerobic co-digestion of dewatered sludge (DS) and food waste (FW) in comparison with mono digestions were investigated. System stability was improved in co-digestion systems with co-substrate acting as a diluting agent to toxic chemicals like ammonia or Na(+). For high-solids digestion of DS, the addition of FW not only improved system stability but also greatly enhanced volumetric biogas production. For high-solids digestion of FW, the addition of DS could reduce Na(+) concentration and help maintain satisfactory stability during the conversion of FW into biogas. System performances of co-digestion systems were mainly determined by the mixing ratios of DS and FW. Biogas production and volatile solids (VSs) reduction in digestion of the co-mixture of DS and FW increased linearly with higher ratios of FW. A kinetic model, which aimed to forecast the performance of co-digestion and to assist reactor design, was developed from long-term semi-continuous experiments. Maximum VS reduction for DS and FW was estimated to be 44.3% and 90.3%, respectively, and first order constant k was found to be 0.17d(-1) and 0.50 d(-1), respectively. Experimental data of co-digestion were in good conformity to the predictions of the model. Copyright © 2012 Elsevier Ltd. All rights reserved.

Environmental assessment of digestate treatment technologies using LCA methodology.

PubMed

Vázquez-Rowe, Ian; Golkowska, Katarzyna; Lebuf, Viooltje; Vaneeckhaute, Céline; Michels, Evi; Meers, Erik; Benetto, Enrico; Koster, Daniel

2015-09-01

The production of biogas from energy crops, organic waste and manure has augmented considerably the amounts of digestate available in Flanders. This has pushed authorities to steadily introduce legislative changes to promote its use as a fertilising agent. There is limited arable land in Flanders, which entails that digestate has to compete with animal manure to be spread. This forces many anaerobic digestion plants to further treat digestate in such a way that it can either be exported or the nitrogen be removed. Nevertheless, the environmental impact of these treatment options is still widely unknown, as well as the influence of these impacts on the sustainability of Flemish anaerobic digestion plants in comparison to other regions where spreading of raw digestate is allowed. Despite important economic aspects that must be considered, the use of Life Cycle Assessment (LCA) is suggested in this study to identify the environmental impacts of spreading digestate directly as compared to four different treatment technologies. Results suggest relevant environmental gains when the digestate mix is treated using the examined conversion technologies prior to spreading, although important trade-offs between impact categories were observed and discussed. The promising results of digestate conversion technologies suggest that further LCA analyses should be performed to delve into, for instance, the appropriateness to shift to nutrient recovery technologies rather than digestate conversion treatments. Copyright © 2015 Elsevier Ltd. All rights reserved.

Enhanced dewaterability of sludge during anaerobic digestion with thermal hydrolysis pretreatment: New insights through structure evolution.

PubMed

Zhang, Jingsi; Li, Ning; Dai, Xiaohu; Tao, Wenquan; Jenkinson, Ian R; Li, Zhuo

2017-12-19

Comprehensive insights into the sludge digestate dewaterability were gained through porous network structure of sludge. We measured the evolution of digestate dewaterability, represented by the solid content of centrifugally dewatered cake, in high-solids sequencing batch digesters with and without thermal hydrolysis pretreatment (THP). The results show that the dewaterability of the sludge after digestion was improved by 3.5% (±0.5%) for unpretreated sludge and 5.1% (±0.4%) for thermally hydrolyzed sludge. Compared to the unpretreated sludge digestate, thermal hydrolysis pretreatment eventually resulted in an improvement of dewaterability by 4.6% (±0.5%). Smaller particle size and larger surface area of sludge were induced by thermal hydrolysis and anaerobic digestion treatments. The structure strength and compactness of sludge, represented by elastic modulus and fractal dimension respectively, decreased with increase of digestion time. The porous network structure was broken up by thermal hydrolysis pretreatment and was further weakened during anaerobic digestion, which correspondingly improved the dewaterability of digestates. The logarithm of elastic modulus increased linearly with fractal dimension regardless of the pretreatment. Both fractal dimension and elastic modulus showed linear relationship with dewaterability. The rheological characterization combined with the analysis of fractal dimension of sewage sludge porous network structure was found applicable in quantitative evaluation of sludge dewaterability, which depended positively on both thermal hydrolysis and anaerobic digestion. Copyright © 2017 Elsevier Ltd. All rights reserved.

The effect of trace element addition to mono-digestion of grass silage at high organic loading rates.

PubMed

Wall, David M; Allen, Eoin; Straccialini, Barbara; O'Kiely, Padraig; Murphy, Jerry D

2014-11-01

This study investigated the effect of trace element addition to mono-digestion of grass silage at high organic loading rates. Two continuous reactors were compared. The first mono-digested grass silage whilst the second operated in co-digestion, 80% grass silage with 20% dairy slurry (VS basis). The reactors were run for 65weeks with a further 5weeks taken for trace element supplementation for the mono-digestion of grass silage. The co-digestion reactor reported a higher biomethane efficiency (1.01) than mono-digestion (0.90) at an OLR of 4.0kgVSm(-3)d(-1) prior to addition of trace elements. Addition of cobalt, iron and nickel, led to an increase in the SMY in mono-digestion of grass silage by 12% to 404LCH4kg(-1)VS and attained a biomethane efficiency of 1.01. Copyright © 2014 Elsevier Ltd. All rights reserved.

The microstructure of starchy food modulates its digestibility.

PubMed

Tian, Jinhu; Ogawa, Yukiharu; Shi, John; Chen, Shiguo; Zhang, Huiling; Liu, Donghong; Ye, Xingqian

2018-06-05

Starch is the main carbohydrate in human nutrition and shows a range of desired food properties. It has been demonstrated that fast digestion of starchy food can induce many health issues (e.g., hyperglycaemia, diabetes, etc.); therefore, how to modulate its digestion is an interesting topic. Previous studies have revealed that the microstructure and digestibility of starchy food of different botanical origin or from multiple processes are quite different; modulating starch digestion by retaining or altering its microstructure may be effective. In the present review, the current knowledge of the relationship between microstructural changes to starchy food and its digestibility at molecular, cell and tissue, and food processing levels is summarized. New technologies focused on microstructure studies and ways to manipulate food microstructure to modulate starch digestibility are also reviewed. In particular, some insights focusing on the future study of microstructure and the digestibility of starchy food are also suggested.

Gross and true ileal digestible amino acid contents of several animal body proteins and their hydrolysates.

PubMed

Cui, J; Chong, B; Rutherfurd, S M; Wilkinson, B; Singh, H; Moughan, P J

2013-07-01

Amino acid compositions of ovine muscle, ovine myofibrillar protein, ovine spleen, ovine liver, bovine blood plasma, bovine blood globulins and bovine serum albumin and the amino acid compositions and in vivo (laboratory rat) true ileal amino acid digestibilities of hydrolysates (sequential hydrolysis with Neutrase, Alcalase and Flavourzyme) of these protein sources were determined. True ileal amino acid digestibility differed (P<0.05) among the seven protein hydrolysates. The ovine myofibrillar protein and liver hydrolysates were the most digestible, with a mean true ileal digestibility across all amino acids of 99%. The least digestible protein hydrolysate was bovine serum albumin with a comparable mean true ileal digestibility of 93%. When the digestible amino acid contents were expressed as proportions relative to lysine, considerable differences, across the diverse protein sources, were found in the pattern of predicted absorbed amino acids. Copyright © 2013 Elsevier Ltd. All rights reserved.

Relationship between body weight and level of fat supplementation on fatty acid digestion in feedlot cattle.

PubMed

Plascencia, A; Mendoza, G D; Vásquez, C; Zinn, R A

2003-11-01

Eight Holstein steers with cannulas in the rumen and proximal duodenum were used in a split-plot design experiment to evaluate the interaction of body weight (175 vs. 370 kg) and level of fat supplementation (0, 3, 6, and 9% yellow grease) on characteristics of digestion and feeding value of fat in finishing diets. Dry matter intake was restricted to 2% of BW. There were no interactions between BW and level of fat supplementation (P > 0.10) on ruminal or total-tract digestion. Level of supplemental fat decreased (linear, P < 0.01) ruminal digestion of OM and NDF, and increased (linear, P < 0.05) ruminal N efficiency. There were no treatment effects (P > 0.10) on postruminal digestion of OM, NDF, and N. There tended to be an interaction (P < 0.10) between BW and level of fat supplementation on postruminal starch digestion. Increasing level of fat supplementation increased postruminal digestion of starch in heavier steers but did not affect starch digestion in lighter steers. There were no interactions (P > 0.10) between BW and level of fat supplementation on postruminal fatty acid digestion. Increasing level of fat supplementation decreased (linear, P < 0.01) postruminal fatty acid digestion, which was due to a decreased (linear, P < 0.01) postruminal digestion of C16:0 and C18:0. Supplemental fat decreased (linear, P < 0.01) total-tract digestion of OM and NDF. The estimated NEm (Mcal/kg) of yellow grease averaged (linear, P < 0.01) 6.02, 5.70, and 5.06 for the 3, 6, and 9% of level supplementation, respectively. We conclude that intestinal fatty acid digestion (FAD, %) is a predictable function (r2 = 0.89; P < 0.01) of total fatty acid intake per unit body weight (FAI, g/kg BW): FAD = 87.560 - 8.591FAI. Depressions in fatty acid digestion with increasing level of intake were due primarily to decreased intestinal absorption of palmitic and stearic acid. Level of fatty acids intake did not appreciably affect intestinal absorption of unsaturated fatty acid. Changes in intestinal fatty acid digestion accounted for most of the variation in the NE value of supplemental fat.

Prediction of in vivo neutral detergent fiber digestibility and digestion rate of potentially digestible neutral detergent fiber: comparison of models.

PubMed

Huhtanen, P; Seppälä, A; Ahvenjärvi, S; Rinne, M

2008-10-01

Eleven 1-pool, seven 2-pool, and three 3-pool models were compared in fitting gas production data and predicting in vivo NDF digestibility and effective first-order digestion rate of potentially digestible NDF (pdNDF). Isolated NDF from 15 grass silages harvested at different stages of maturity was incubated in triplicate in rumen fluid-buffer solution for 72 h to estimate the digestion kinetics from cumulative gas production profiles. In vivo digestibility was estimated by the total fecal collection method in sheep fed at a maintenance level of feeding. The concentration of pdNDF was estimated by a 12-d in situ incubation. The parameter values from gas production profiles and pdNDF were used in a 2-compartment rumen model to predict pdNDF digestibility using 50 h of rumen residence time distributed in a ratio of 0.4:0.6 between the non-escapable and escapable pools. The effective first-order digestion rate was computed both from observed in vivo and model-predicted pdNDF digestibility assuming the passage kinetic model described above. There were marked differences between the models in fitting the gas production data. The fit improved with increasing number of pools, suggesting that silage pdNDF is not a homogenous substrate. Generally, the models predicted in vivo NDF digestibility and digestion rate accurately. However, a good fit of gas production data was not necessarily translated into improved predictions of the in vivo data. The models overestimating the asymptotic gas volumes tended to underestimate the in vivo digestibility. Investigating the time-related residuals during the later phases of fermentation is important when the data are used to estimate the first-order digestion rate of pdNDF. Relatively simple models such as the France model or even a single exponential model with discrete lag period satisfied the minimum criteria for a good model. Further, the comparison of feedstuffs on the basis of parameter values is more unequivocal than in the case of multiple-pool models.

Measurement of protein digestibility in humans by a dual-tracer method.

PubMed

Devi, Sarita; Varkey, Aneesia; Sheshshayee, M S; Preston, Thomas; Kurpad, Anura V

2018-06-01

Recent evaluations of the risk of dietary protein deficiency have indicated that protein digestibility may be a key limiting factor in the provision of indispensable amino acids (IAAs), particularly for vulnerable populations living in challenging environments where intestinal dysfunction may exist. Since the digestion of protein occurs only in the small intestine, and the metabolic activity of colonic bacteria confounds measurements at the fecal level, there is a need to develop noninvasive protein digestibility measurements at the ileal level. We used a dual-tracer method with stable isotopes to characterize the digestibility of uniformly labeled [13C]-spirulina protein as a standard protein, in comparison to a mixture of 2H-labeled crystalline amino acids, and then demonstrated the use of this standard protein to measure the digestibility of selected legumes (chick pea and mung bean) through the use of proteins that were intrinsically labeled with 2H. The digestibility of uniformly labeled [13C]-spirulina was first measured in 6 healthy volunteers (3 males and 3 females) by feeding it along with a standard mixture of 2H-labeled amino acids, in a dual-tracer, plateau-fed test meal approach. Next, intrinsically labeled legume protein digestibility was studied with a similar dual-tracer approach, with uniformly labeled [13C]-spirulina as the standard, when processed differently before consumption. The average digestibility of IAA in spirulina protein was 85.2%. The average IAA digestibility of intrinsically 2H-labeled chick pea and mung bean protein was 56.6% and 57.7%, respectively. Dehulling of mung bean before ingestion increased the average IAA digestibility by 9.9% in comparison to whole mung bean digestibility. An innovative, minimally invasive "dual-stable-isotope" method was developed to measure protein digestibility, in which the ingestion of an intrinsically 2H-labeled test protein along with a 13C-labeled standard protein of known digestibility allows for an accurate measure of digestion and absorption of the intrinsically labeled protein. This minimally invasive method is critical to redefining protein quality and will aid in revisiting human protein requirements in different settings and in vulnerable populations. This trial was registered at Clinical Trials Registry-India as CTRI/2017/11/010468.

Changes in chemical composition and digestibility of three maize stover components digested by white-rot fungi.

PubMed

Lynch, J P; O'Kiely, P; Murphy, R; Doyle, E M

2014-08-01

Maize stover (total stem and leaves) is not considered a ruminant feed of high nutritive value. Therefore, an improvement in its digestibility may increase the viability of total forage maize production systems in marginal growth regions. The objective of this study was to describe the changes in chemical composition during the storage of contrasting components of maize stover (leaf, upper stem and lower stem) treated with either of two lignin degrading white-rot fungi (WRF; Pleurotus ostreatus, Trametes versicolor). Three components of maize stover (leaf, upper stem and lower stem), harvested at a conventional maturity for silage production, were digested with either of two WRF for one of four digestion durations (1-4 months). Samples taken prior to fungal inoculation were used to benchmark the changes that occurred. The degradation of acid detergent lignin was observed in all sample types digested with P. ostreatus; however, the loss of digestible substrate in all samples inoculated with P. ostreatus was high, and therefore, P. ostreatus-digested samples had a lower dry matter digestibility than samples prior to inoculation. Similarly, T. veriscolor-digested leaf underwent a non-selective degradation of the rumen-digestible components of fibre. The changes in chemical composition of leaf, upper stem and lower stem digested with either P. ostreatus or T. veriscolor were not beneficial to the feed value of the forage, and incurred high DM losses. Journal of Animal Physiology and Animal Nutrition © 2013 Blackwell Verlag GmbH.

Aged-engineered nanoparticles effect on sludge anaerobic digestion performance and associated microbial communities.

PubMed

Eduok, Samuel; Ferguson, Robert; Jefferson, Bruce; Villa, Raffaella; Coulon, Frédéric

2017-12-31

To investigate the potential effect of aged engineered nanoparticles (a-ENPs) on sludge digestion performance, 150L pilot anaerobic digesters (AD) were fed with a blend of primary and waste activated sludge spiked either with a mixture of silver oxide, titanium dioxide and zinc oxide or a mixture of their equivalent bulk metal salts to achieve a target concentration of 250, 2000, and 2800mgkg -1 dry weight, respectively. Volatile fatty acids (VFA) were 1.2 times higher in the spiked digesters and significantly different (p=0.05) from the control conditions. Specifically, isovaleric acid concentration was 2 times lower in the control digester compared to the spiked digesters, whereas hydrogen sulfide was 2 times lower in the ENPs spiked digester indicating inhibitory effect on sulfate reducing microorganisms. Based on the ether-linked isoprenoids concentration, the total abundance of methanogens was 1.4 times lower in the ENPs spiked digester than in the control and metal salt spiked digesters. Pyrosequencing indicated 80% decrease in abundance and diversity of methanogens in ENPs spiked digester compared to the control digester. Methanosarcina acetivorans and Methanosarcina barkeri were identified as nano-tolerant as their relative abundance increased by a factor of 6 and 11, respectively, compared to the other digesters. The results further provide compelling evidence on the resilience of Fusobacteria, Actinobacteria and the Trojan horse-like effect of ENPs which offered a competitive advantage to some organisms while reducing microbial abundance and diversity. Copyright © 2017 The Authors. Published by Elsevier B.V. All rights reserved.

Microalgal Cultivation in Treating Liquid Digestate from Biogas Systems.

PubMed

Xia, Ao; Murphy, Jerry D

2016-04-01

Biogas production via anaerobic digestion (AD) has rapidly developed in recent years. In addition to biogas, digestate is an important byproduct. Liquid digestate is the major fraction of digestate and may contain high levels of ammonia nitrogen. Traditional processing technologies (such as land application) require significant energy inputs and raise environmental risks (such as eutrophication). Alternatively, microalgae can efficiently remove the nutrients from digestate while producing high-value biomass that can be used for the production of biochemicals and biofuels. Both inorganic and organic carbon sources derived from biogas production can significantly improve microalgal production. Land requirement for microalgal cultivation is estimated as 3% of traditional direct land application of digestate. Copyright © 2015 Elsevier Ltd. All rights reserved.

Temperature-phased anaerobic digestion of food waste: A comparison with single-stage digestions based on performance and energy balance.

PubMed

Xiao, Benyi; Qin, Yu; Zhang, Wenzhe; Wu, Jing; Qiang, Hong; Liu, Junxin; Li, Yu-You

2018-02-01

The temperature-phased anaerobic digestion (TPAD) of food waste was studied for the purpose of comparing with single-stage mesophilic and thermophilic anaerobic digestion. The biogas and methane yields in the TPAD during the steady period were 0.759 ± 0.115 L/g added VS and 0.454 ± 0.201 L/g added VS, which were lower than those in the two single-stage anaerobic digestion. The improper sludge retention time may be the reason for the lower biogas and methane production in TPAD. The removal of volatile solids in the TPAD was 78.55 ± 4.59% and the lowest among the three anaerobic digestion processes. The reaction ratios of the four anaerobic digestion steps in the TPAD were all lower than those in the two single-stage anaerobic digestion. The energy conversion efficiency of the degraded substrate in the TPAD was similar with those in single-stage mesophilic and thermophilic anaerobic digestion systems. Copyright © 2017 Elsevier Ltd. All rights reserved.

Nucleic acids digestion by enzymes in the stomach of snakehead (Channa argus) and banded grouper (Epinephelus awoara).

PubMed

Liu, Yu; Zhang, Yanfang; Jiang, Wei; Wang, Jing; Pan, Xiaoming; Wu, Wei; Cao, Minjie; Dong, Ping; Liang, Xingguo

2017-02-01

Dietary nucleic acids (NAs) were important nutrients. However, the digestion of NAs in stomach has not been studied. In this study, the digestion of NAs by enzymes from fish stomach was investigated. The snakehead pepsins (SP) which were the main enzymes in stomach were extracted and purified. The purity of SP was evaluated by SDS-PAGE and HPLC. The snakehead pepsin 2 (SP2) which was the main component in the extracts was used for investigating the protein and NAs digestion activity. SP2 could digest NAs, including λ DNA and salmon sperm DNA. Interestingly, the digestion could be inhibited by treatment of alkaline solution at pH 8.0 and pepstatin A, and the digestion could happen either in the presence or absence of hemoglobin (Hb) and BSA as the protein substrates. Similarly, the stomach enzymes of banded grouper also showed the NAs digestion activity. NAs could be digested by the stomach enzymes of snakehead and banded grouper. It may be helpful for understanding both animal nutrition and NAs metabolic pathway.

Digestibility of fucosylated glycosaminoglycan from sea cucumber and its effects on digestive enzymes under simulated salivary and gastrointestinal conditions.

PubMed

Zhao, Longyan; Qin, Yujing; Guan, Ruowei; Zheng, Wenqi; Liu, Jikai; Zhao, Jinhua

2018-04-15

The digestibility of fucosylated glycosaminoglycan (FG) and its effects on digestive enzymes were investigated using an in vitro digestion model. Results showed that the molecular weight and the reducing sugar content of FG were not significantly changed, and no free monosaccharides released from FG were detected after the salivary, gastric and intestinal digestion, indicating that both the backbone and the sulfated fucose branches of FG are resistant to be cleaved in the saliva and gastrointestinal tract environments. Furthermore, FG can dose-dependently inhibit digestive enzymes such as α-amylase, pepsin and pancreatic lipase in different degrees under the simulated digestion conditions due to the sulfate and carboxyl groups. These physiological effects of FG may help control the postprandial glucose concentration and have the potential in the prevention or treatment of reflux disease and obesity. The findings may provide information on the digestibility and beneficial physiological effects of FG as a potential natural product to promote human health. Copyright © 2018 Elsevier Ltd. All rights reserved.

Anaerobic co-digestion of cyanide containing cassava pulp with pig manure.

PubMed

Glanpracha, Naraporn; Annachhatre, Ajit P

2016-08-01

Anaerobic co-digestion of cyanide-containing cassava pulp with pig manure was evaluated using laboratory scale mesophilic digester. The digester was operated in a semi-continuous mode with the mixed feedstock having C/N ratio of 35:1. Digester startup was accomplished in 60days with loading of 0.5-1kgVS/m(3)d. Subsequently, the loading to digester was increased step-wise from 2 to 9kgVS/m(3)d. Digester performance was stable at loading between 2 and 6kgVS/m(3)d with an average volatile solid removal and methane yield of 82% and 0.38m(3)/kgVSadded, respectively. However, beyond loading of 7kgVS/m(3)d, solubilization of particulate matter did not take place efficiently. Cyanide present in cassava pulp was successfully degraded indicating that anaerobic sludge in the digester was well acclimatized to cyanide. The results show that cassava pulp can be successfully digested anaerobically with pig manure as co-substrate without any inhibitory effect of cyanide present in the cassava pulp. Copyright © 2016 Elsevier Ltd. All rights reserved.

Enhancing post aerobic digestion of full-scale anaerobically digested sludge using free nitrous acid pretreatment.

PubMed

Wang, Qilin; Zhou, Xu; Peng, Lai; Wang, Dongbo; Xie, Guo-Jun; Yuan, Zhiguo

2016-05-01

Post aerobic digestion of anaerobically digested sludge (ADS) has been extensively applied to the wastewater treatment plants to enhance sludge reduction. However, the degradation of ADS in the post aerobic digester itself is still limited. In this work, an innovative free nitrous acid (HNO2 or FNA)-based pretreatment approach is proposed to improve full-scale ADS degradation in post aerobic digester. The post aerobic digestion was conducted by using an activated sludge to aerobically digest ADS for 4 days. Degradations of the FNA-treated (treated at 1.0 and 2.0 mg N/L for 24 h) and untreated ADSs were then determined and compared. The ADS was degraded by 26% and 32%, respectively, in the 4-day post aerobic digestion period while being pretreated at 1.0 and 2.0 mg HNO2-N/L. In comparison, only 20% of the untreated ADS was degraded. Economic analysis demonstrated that the implementation of FNA pretreatment can be economically favourable or not depending on the sludge transport and disposal cost. Copyright © 2016 Elsevier Ltd. All rights reserved.

Comparison of sludge digestion under aerobic and anaerobic conditions with a focus on the degradation of proteins at mesophilic temperature.

PubMed

Shao, Liming; Wang, Tianfeng; Li, Tianshui; Lü, Fan; He, Pinjing

2013-07-01

Aerobic and anaerobic digestion are popular methods for the treatment of waste activated sludge. However, the differences in degradation of sludge during aerobic and anaerobic digestion remain unclear. In this study, the sludge degradation during aerobic and anaerobic digestion was investigated at mesophilic temperature, focused on protein based on the degradation efficiency and degree of humification. The duration of aerobic and anaerobic digestion was about 90 days. The final degradation efficiency of volatile solid was 66.1 ± 1.6% and 66.4 ± 2.4% under aerobic and anaerobic conditions, respectively. The final degradation efficiency of protein was 67.5 ± 1.4% and 65.1 ± 2.6% under aerobic and anaerobic conditions, respectively. The degradation models of volatile solids were consistent with those of protein under both aerobic and anaerobic conditions. The solubility of protein under aerobic digestion was greater than that under anaerobic digestion. Moreover, the humification index of dissolved organic matter of aerobic digestion was greater than that during anaerobic digestion. Copyright © 2013 Elsevier Ltd. All rights reserved.

Nutrient digestibility of broiler feeds containing different levels of variously processed rice bran stored for different periods.

PubMed

Mujahid, A; Asif, M; ul Haq, I; Abdullah, M; Gilani, A H

2003-09-01

Nutrient digestibility of broiler feeds containing different levels of variously processed rice bran stored for varying periods was determined. A total of 444 Hubbard male chicks were used to conduct four trials. Each trial was carried out on 111 chicks to determine digestibility of 36 different feeds. Chicks of 5 wk age were fed feeds containing raw, roasted, and extruded rice bran treated with antioxidant, Bianox Dry (0, 125, 250 g/ton), stored for a periods of 0, 4, 8, and 12 mo and used at levels of 0, 10, 20, and 30% in feeds. Digestibility coefficients for fat and fiber of feeds were determined. Increasing storage periods of rice bran significantly reduced the fat digestibility of feed, whereas no difference in fiber digestibility was observed. Processing of rice bran by extrusion cooking significantly increased digestibility of fat even used at higher levels in broiler feeds. Interaction of storage, processing, and levels was significant for fat digestibility. Treatments of rice bran by different levels of antioxidant had no effect on digestibility of fat and fiber when incorporated in broiler feed.

Digestibility of Bovine Serum Albumin and Peptidomics of the Digests: Effect of Glycation Derived from α-Dicarbonyl Compounds.

PubMed

Sheng, Bulei; Larsen, Lotte Bach; Le, Thao T; Zhao, Di

2018-03-21

α-Dicarbonyl compounds, which are widely generated during sugar fragmentation and oil oxidation, are important precursors of advanced glycation end products (AGEs). In this study, the effect of glycation derived from glyoxal (GO), methylglyoxal (MGO) and diacetyl (DA) on the in vitro digestibility of bovine serum albumin (BSA) was investigated. Glycation from α-dicarbonyl compounds reduced digestibility of BSA in both gastric and intestinal stage of digestion according to measurement of degree of hydrolysis. Changes in peptide composition of digests induced by glycation were displayed, showing absence of peptides, occurrence of new peptides and formation of peptide-AGEs, based on the results obtained using liquid chromatography electron-spray-ionization tandem mass spectrometry (LC-ESI-MS/MS). Crosslinked glycation structures derived from DA largely reduced the sensitivity of glycated BSA towards digestive proteases based on sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) results. Network structures were found to remain in the digests of glycated samples by transmission electron microscope (TEM), thus the impact of AGEs in unabsorbed digests on the gut flora should be an interest for further studies.

DOE Office of Scientific and Technical Information (OSTI.GOV)

Shi, Jinshu; Shi, Sheldon Q.; Barnes, H. Michael

The objective of this study was to develop a hermetical alkali digestion process to obtain single cellulosic fibers from kenaf bast. Kenaf bast were hermetically digested into single fiber using a 5% sodium hydroxide solution for one hour at four different temperatures (80 ° C, 110 ° C, 130 ° C, and 160 ° C). The hermetical digestion process used in this study produced fibers with high cellulose content (84.2–92.3%) due to the removal of lignin and hemicelluloses. The surface hardness and elastic modulus of the fibers digested at 130 ° C and 160 ° C were improved significantly comparedmore » with those digested at 80 ° C. The tensile modulus and tensile strength of the individual fibers reduced as the digestion temperature increased from 110 ° C to 160 ° C. Micropores were generated in fiber cell wall when the fibers were digested at 130 ° C and 160 ° C. The studies on the composites that were made from polypropylene reinforced with the digested fibers indicated that the compatibility between the digested fibers and polypropylene matrix was poor.« less

Characterization of Hydrolysis Kinetics in Staged Anaerobic Digestion of Wastewater Treatment Sludge.

PubMed

Zamanzadeh, Mirzaman; Parker, Wayne J

2018-01-01

  The hydrolysis of mixed primary and secondary sludges in two-stage anaerobic digestion was evaluated and compared with conventional single-stage digestion, using various temperature-phased configurations of M1-M2, M1-T3, T1-T2, and T1-M3. A dual hydrolysis model best described the hydrolysis in all tests. This model was also able to consistently estimate the readily and slowly fractions of particulate chemical oxygen demand (COD) of raw sludge used in the tests. The hydrolysis kinetic coefficients (Khyd_s and Khyd_r) estimated for the mesophilic digesters were significantly greater in the short hydraulic retention time (HRT) M1 digester than those of the extended HRT digesters. Conversely, at thermophilic temperatures only Khyd_r was greater in short HRT T1 digester when compared to the extended HRT digesters. The increased Khyd_r and reduced Khyd_s values due to staging effect were explained with surface reaction models and endogenous decay. The temperature dependency of Khyd_s and Khyd_r was also explored in the staged digesters.

Accelerated digestion of nucleic acids by pepsin from the stomach of chicken.

PubMed

Liu, Y; Zhang, Y; Guo, H; Wu, W; Dong, P; Liang, X

2016-10-01

Nucleic acids have become an important nutritional supplement in poultry feed; however, the digestion of nucleic acids in poultry is unclear. The objective of this study was to investigate the digestion of nucleic acids by chicken pepsin in vitro. The extracted pepsinogen from the stomach of the chicken was purified to homogeneity. Upon activation at pH 2.0, chicken pepsinogen was converted to its active form. Nucleic acids, including λ-DNA, salmon sperm DNA and single-strand DNA (ssDNA), can be used as substrates and digested into short-chain oligonucleotides by pepsin. Interestingly, the digestion of the nucleic acids was inhibited when pepsin was treated by alkaline solution (pH 8.0) or pepstatin A. Also, the digestion of the nucleic acids was not affected by the addition of haemoglobin or bovine serum albumin. The results suggested that nucleic acids could be digested by chicken pepsin. Thus pepsin may have a role in digesting nucleic acids in vivo. Nucleic acids added to poultry fed may be digested, starting from the stomach.

Identification and Relative Quantification of Bioactive Peptides Sequentially Released during Simulated Gastrointestinal Digestion of Commercial Kefir.

PubMed

Liu, Yufang; Pischetsrieder, Monika

2017-03-08

Health-promoting effects of kefir may be partially caused by bioactive peptides. To evaluate their formation or degradation during gastrointestinal digestion, we monitored changes of the peptide profile in a model of (1) oral, (2) gastric, and (3) small intestinal digestion of kefir. Matrix-assisted laser desorption/ionization time-of-flight mass spectroscopy analyses revealed clearly different profiles between digests 2/3 and kefir/digest 1. Subsequent ultraperformance liquid chromatography-electrospray ionization-tandem mass spectrometry identified 92 peptides in total (25, 25, 43, and 30, partly overlapping in kefir and digests 1, 2, and 3, respectively), including 16 peptides with ascribed bioactivity. Relative quantification in scheduled multiple reaction monitoring mode showed that many bioactive peptides were released by simulated digestion. Most prominently, the concentration of angiotensin-converting enzyme inhibitor β-casein 203-209 increased approximately 10 000-fold after combined oral, gastric, and intestinal digestion. Thus, physiological digestive processes may promote bioactive peptide formation from proteins and oligopeptides in kefir. Furthermore, bioactive peptides present in certain compartments of the gastrointestinal tract may exert local physiological effects.

Validation of an in vitro model for predicting rumen and total-tract fiber digestibility in dairy cows fed corn silages with different in vitro neutral detergent fiber digestibilities at 2 levels of dry matter intake.

PubMed

Lopes, F; Cook, D E; Combs, D K

2015-01-01

An in vivo study was performed to validate an in vitro procedure that predicts rate of fiber digestion and total-tract neutral detergent fiber digestibility (TTNDFD). Two corn silages that differed in fiber digestibility were used in this trial. The corn silage with lower fiber digestibility (LFDCS) had the TTNDFD prediction of 36.0% of total NDF, whereas TTNDFD for the corn silage with higher fiber digestibility (HFDCS) was 44.9% of total neutral detergent fiber (NDF). Two diets (1 with LFDCS and 1 with HFDCS) were formulated and analyzed using the in vitro assay to predict the TTNDFD and rumen potentially digestible NDF (pdNDF) digestion rate. Similar diets were fed to 8 ruminally cannulated, multiparous, high-producing dairy cows in 2 replicated 4×4 Latin squares with 21-d periods. A 2×2 factorial arrangement of treatments was used with main effects of intake (restricted to approximately 90% of ad libitum intake vs. ad libitum) and corn silage of different fiber digestibility. Treatments were restricted and ad libitum LFDCS as well as restricted and ad libitum HFDCS. The input and output values predicted from the in vitro model were compared with in vivo measurements. The pdNDF intake predicted by the in vitro model was similar to pdNDF intake observed in vivo. Also, the pdNDF digestion rate predicted in vitro was similar to what was observed in vivo. The in vitro method predicted TTNDFD of 50.2% for HFDCS and 42.9% for LFDCS as a percentage of total NDF in the diets, whereas the in vivo measurements of TTNDFD averaged 50.3 and 48.6% of total NDF for the HFDCS and LFDCS diets, respectively. The in vitro TTNDFD assay predicted total-tract NDF digestibility of HFDCS diets similar to the digestibility observed in vivo, but for LFDCS diets the assay underestimated the digestibility compared with in vivo. When the in vitro and in vivo measurements were compared without intake effect (ad libitum and restricted) considering only diet effect of silage fiber digestibility (HFDCS and LFDCS), no differences were observed between methods. These values suggest that our in vitro TTNDFD model could be used to predicted rate of fiber digestion and NDF digestibility for dairy cattle. Copyright © 2015 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Structural aspects of digestion of medium chain triglycerides studied in real time using sSAXS and Cryo-TEM.

PubMed

Phan, Stephanie; Hawley, Adrian; Mulet, Xavier; Waddington, Lynne; Prestidge, Clive A; Boyd, Ben J

2013-12-01

The purpose of this study was to investigate the colloidal structures formed on digestion of medium chain triglyceride (MCT) with a specific objective of identifying and characterizing a previously reported vesicular phase, which has been linked to supersaturation and anomalous digestion kinetics, and to evaluate the influence of lipid mass and enzyme inhibition on self assembled structure. MCT was digested in vitro and nanostructure was monitored in real time using synchrotron small angle X-ray scattering (sSAXS), and morphology was studied using cryogenic transmission electron microscopy (cryo-TEM). Formation of the putative vesicular phase formed on digestion of MCT was confirmed and its structural attributes were determined. Vesicle formation was dependent on lipid mass and bile salt concentration. The use of enzyme inhibitor for offline analysis of lipolysis samples did influence structural aspects of the digestion medium when compared to real time evaluation. The formation of a vesicular phase was directly linked to the kinetics of lipid digestion. Vesicle formation is linked to lipid mass, or more specifically the ratio of lipid to bile salts present in the digestion mixture. Inhibition of lipase to halt digestion during sampling for offline analysis must be done with caution as structural aspects were shown to differ for the MCT digests with and without inhibitor present.

The challenges of anaerobic digestion and the role of biochar in optimizing anaerobic digestion.

PubMed

Fagbohungbe, Michael O; Herbert, Ben M J; Hurst, Lois; Ibeto, Cynthia N; Li, Hong; Usmani, Shams Q; Semple, Kirk T

2017-03-01

Biochar, like most other adsorbents, is a carbonaceous material, which is formed from the combustion of plant materials, in low-zero oxygen conditions and results in a material, which has the capacity to sorb chemicals onto its surfaces. Currently, research is being carried out to investigate the relevance of biochar in improving the soil ecosystem, digestate quality and most recently the anaerobic digestion process. Anaerobic digestion (AD) of organic substrates provides both a sustainable source of energy and a digestate with the potential to enhance plant growth and soil health. In order to ensure that these benefits are realised, the anaerobic digestion system must be optimized for process stability and high nutrient retention capacity in the digestate produced. Substrate-induced inhibition is a major issue, which can disrupt the stable functioning of the AD system reducing microbial breakdown of the organic waste and formation of methane, which in turn reduces energy output. Likewise, the spreading of digestate on land can often result in nutrient loss, surface runoff and leaching. This review will examine substrate inhibition and their impact on anaerobic digestion, nutrient leaching and their environmental implications, the properties and functionality of biochar material in counteracting these challenges. Copyright © 2016 Elsevier Ltd. All rights reserved.

Structure and function of the digestive system of solen grandis dunker

NASA Astrophysics Data System (ADS)

Sheng, Xiuzhen; Zhan, Wenbin; Ren, Sulian

2003-10-01

Structure and function of the digestive system of a bivalve mollusc, Solen grandis, were studied using light microscopy and histochemical methods. The wall of digestive tube consists of four layers: the mucosal epithelium, connective tissue, muscular and fibrosa or serosa (only in the portion of rectum) from the inner to the outer. The ciliated columnar epithelial cells, dispersed by cup-shaped mucous cells, rest on a thin base membrane. There are abundant blood spaces in connective tissue layer. The digestive diverticula are composed of multi-branched duct and digestive tubules. The digestive tubules are lined with digestive and basophilic secretory cells, and surrounded by a layer of smooth muscle fibers and connective tissues. Activities of acid and alkaline phosphatases, esterase and lipase are detected in the digestive cells, and the epithelia of stomach and intestine, suggesting that these cells are capable of intracellular digesting of food materials and absorbing. Besides, acid phosphatase and esterase activities are present in the posterior portion of esophagus. Phagocytes are abundant in blood spaces and the lumens of stomach and intestine, containing brown granules derived from the engulfed food materials. The present work indicates that phagocytes play important roles in ingestion and digestion of food materials, which is supported as well by the activities of acid phosphatase, esterase and lipase detected in blood spaces.

Anaerobically digested food waste in compost for Agaricus bisporus and Agaricus subrufescens and its effect on mushroom productivity.

PubMed

Stoknes, Ketil; Beyer, David M; Norgaard, Erik

2013-07-01

Source-separated food waste is increasingly being treated by means of hygienisation followed by anaerobic digestion. The fibrous digester residue (digestate) is a potential mushroom substrate, while heat from the biogas can provide steam for the cultivation process. Using bag experiments the present study explored digestate as a full substitute for chicken manure conventionally used in mushroom composts. After mixing, a rapid temperature development in the compost was stimulated by a small amount of chicken manure, as aerobic microbial seeding. Mechanical elimination of lumps was essential for full mycelial colonisation. Three straw digestate composts had Agaricus bisporus mushroom yields above 370 g kg⁻¹ substrate. The optimal compost water content was 600 g kg⁻¹ at inoculation, and high digestate content (up to 500 g kg⁻¹ by dry weight) did not affect yield for this species. High yields of A. subrufescens (200 g kg⁻¹) were related to drier composts of lower digestate content (more straw) and lower pH values at inoculation. Digestate successfully substituted chicken manure in straw composts without affecting mushroom yields for both species. There were no clear differences between straw digestate and control composts in terms of mushroom dry matter, size, nitrogen or ash content. © 2012 Society of Chemical Industry.

Protein oxidation and proteolysis during roasting and in vitro digestion of fish (Acipenser gueldenstaedtii).

PubMed

Hu, Lyulin; Ren, Sijie; Shen, Qing; Ye, Xingqian; Chen, Jianchu; Ling, Jiangang

2018-04-15

Roasted fish enjoys great popularity in Asia, but how roasting and subsequent digestion influence the oxidation and proteolysis of fish meat is unknown. This paper is aimed to investigate the effect of roasting time on lipid and protein oxidation and their evolution and consequence on proteolysis during simulated digestion of fish fillets. Several oxidation markers (TBARS, free thiols, total carbonyls and Schiff bases) were employed to assess the oxidation of fish. SDS-PAGE and TBNS assay for free amino groups were used to study the proteolysis during gastrointestinal digestion. The results showed that significant lipid and protein oxidative changes occurring in roasted fish fillets were reinforced after gastric digestion and were much more intense after intestinal digestion. Throughout the roasting and digestion, close interconnection between lipid and protein was also manifested as the levels of total carbonyls and Schiff bases rose while TBARS fell. Furthermore, free amino groups decreased with prolonged roasting time, signifying protein oxidation before digestion resulted in impaired proteolysis during digestion. This paper indicated the lipid and protein oxidation of fish fillets could be dependent on time of roasting, and the oxidation continued to develop and have an impact on proteolysis during in vitro digestion. This article is protected by copyright. All rights reserved.

A novel image processing method to determine the nutritional condition of lobsters.

PubMed

Berillis, P; Simon, C; Mente, E; Sofos, F; Karapanagiotidis, I T

2013-02-01

The digestive gland of crustacean is involved in various metabolic activities, including the synthesis and secretion of digestive enzymes that begin the process of food digestion, intracellular digestion and absorption of nutrients, storage of reserves, and disposal of waste products. It consists of two glandular lobes which extensively subdivide to form a complex of blind-ending tubules, whose size, surface area, and digestive cells are associated with intracellular digestion and the nutritional status of the organism. The aim of this paper was to study the morphology of the digestive gland in various lobster species and calculate the surface area of tubules, lumen and digestive cells (R-, F-, and B-cells) and their ratios to total tubule surface area. The similarity in ratios obtained in this study between individual lobsters suggests that the method developed in this study can be successfully applied to a range of species. This study describes a novel image processing algorithm for the automatic measurement of the hepatopancreas structure using stained cross sections of digestive gland tubules. The proposed new methodology could be used for studying the physiology and nutrient metabolism of lobsters and other crustaceans. The computer-aided analysis described in this paper is accurate for the quantitative assessment of the lobster's digestive gland structure. Copyright © 2012 Elsevier Ltd. All rights reserved.

Occurrence and Fate of Trace Contaminants during Aerobic and Anaerobic Sludge Digestion and Dewatering.

PubMed

Guerra, Paula; Kleywegt, Sonya; Payne, Michael; Svoboda, M Lewina; Lee, Hing-Biu; Reiner, Eric; Kolic, Terry; Metcalfe, Chris; Smyth, Shirley Anne

2015-07-01

Digestion of municipal wastewater biosolids is a necessary prerequisite to their beneficial use in land application, in order to protect public health and the receiving environment. In this study, 13 pharmaceuticals and personal care products (PPCPs), 11 musks, and 17 polybrominated diphenyl ethers were analyzed in 84 samples including primary sludge, waste activated sludge, digested biosolids, dewatered biosolids, and dewatering centrate or filtrate collected from five wastewater treatment plants with aerobic or anaerobic digestion. Aerobic digestion processes were sampled during both warm and cold temperatures to analyze seasonal differences. Among the studied compounds, triclosan, triclocarban, galaxolide, and BDE-209 were the substances most frequently detected under different treatment processes at levels up to 30,000 ng/g dry weight. Comparing aerobic and anaerobic digestion, it was observed that the levels of certain PPCPs and musks were significantly higher in anaerobically digested biosolids, relative to the residues from aerobic digestion. Therefore, aerobic digestion has the potential advantage of reducing levels of PPCPs and musks. On the other hand, anaerobic digestion has the advantage of recovering energy from the biosolids in the form of combustible gases while retaining the nutrient and soil conditioning value of this resource. Copyright © by the American Society of Agronomy, Crop Science Society of America, and Soil Science Society of America, Inc.

Optimizing the Logistics of Anaerobic Digestion of Manure

NASA Astrophysics Data System (ADS)

Ghafoori, Emad; Flynn, Peter C.

Electrical power production from the combustion of biogas from anaerobic digestion (AD) of manure is a means of recovering energy from animal waste. We evaluate the lowest cost method of moving material to and from centralized AD plants serving multiple confined feeding operations. Two areas are modeled, Lethbridge County, Alberta, Canada, an area of concentrated beef cattle feedlots, and Red Deer County, Alberta, a mixed-farming area with hog, dairy, chicken and beef cattle farms, and feedlots. We evaluate two types of AD plant: ones that return digestate to the source confined feeding operation for land spreading (current technology), and ones that process digestate to produce solid fertilizer and a dischargeable water stream (technology under development). We evaluate manure and digestate trucking, trucking of manure with return of digestate by pipelines, and pipelining of manure plus digestate. We compare the overall cost of power from these scenarios to farm or feedlot-based AD units. For a centralized AD plant with digestate return for land spreading the most economical transport option for manure plus digestate is by truck for the mixed-farming area and by pipelines for the concentrated feedlot area. For a centralized AD plant with digestate processing, the most economical transport option is trucking of manure for both cases.

Impact of different particle size distributions on anaerobic digestion of the organic fraction of municipal solid waste.

PubMed

Zhang, Y; Banks, C J

2013-02-01

Particle size may significantly affect the speed and stability of anaerobic digestion, and matching the choice of particle size reduction equipment to digester type can thus determine the success or failure of the process. In the current research the organic fraction of municipal solid waste was processed using a combination of a shear shredder, rotary cutter and wet macerator to produce streams with different particle size distributions. The pre-processed waste was used in trials in semi-continuous 'wet' and 'dry' digesters at organic loading rate (OLR) up to 6kg volatile solids (VS) m(-3)day(-1). The results indicated that while difference in the particle size distribution did not change the specific biogas yield, the digester performance was affected. In the 'dry' digesters the finer particle size led to acidification and ultimately to process failure at the highest OLR. In 'wet' digestion a fine particle size led to severe foaming and the process could not be operated above 5kgVSm(-3)day(-1). Although the trial was not designed as a direct comparison between 'wet' and 'dry' digestion, the specific biogas yield of the 'dry' digesters was 90% of that produced by 'wet' digesters fed on the same waste at the same OLR. Copyright © 2012 Elsevier Ltd. All rights reserved.

Digestive oncologist in the gastroenterology training curriculum

PubMed Central

Mulder, Chris Jacob Johan; Peeters, Marc; Cats, Annemieke; Dahele, Anna; Droste, Jochim Terhaar sive

2011-01-01

Until the late 1980s, gastroenterology (GE) was considered a subspecialty of Internal Medicine. Today, GE also incorporates Hepatology. However, Digestive Oncology training is poorly defined in the Hepatogastroenterology (HGE)-curriculum. Therefore, a Digestive Oncology curriculum should be developed and this document might be a starting point for such a curriculum. HGE-specialists are increasingly resisting the paradigm in which they play only a diagnostic and technical role in the management of digestive tumors. We suggest minimum end-points in the standard HGE-curriculum for oncology, and recommend a focus year in the Netherlands for Digestive Oncology in the HGE-curriculum. To produce well-trained digestive oncologists, an advanced Digestive Oncology training program with specific qualifications in Digestive Oncology (2 years) has been developed. The schedule in Belgium includes a period of at least 6 mo to be spent in a medical oncology department. The goal of these programs remains the production of well-trained digestive oncologists. HGE specialists are part of the multidisciplinary oncological teams, and some have been administering chemotherapy in their countries for years. In this article, we provide a road map for the organization of a proper training in Digestive Oncology. We hope that the World Gastroenterology Organisation and other (inter)national societies will support the necessary certifications for this specific training in the HGE-curriculum. PMID:21556128

Microbial release of ferulic and p-coumaric acids from forages and their digestibility in lactating cows fed total mixed rations with different forage combinations.

PubMed

Cao, Bin-Bin; Jin, Xin; Yang, Hong-Jian; Li, Sheng-Li; Jiang, Lin-Shu

2016-01-30

Ferulic acid (FA) and p-coumaric acid (PCA) are widely distributed in graminaceous plant cell walls. This study investigated the in vitro and in vivo digestibility of ester-linked FA (FAest) and PCA (PCAest) in lactating dairy cows. Regarding corn stover, ensiled corn stover, whole corn silage, Chinese wild ryegrass and alfalfa hay with different phenolic acid profiles, the in vitro rumen digestibility of forage FAest and PCAest was negatively correlated with the ether-linked FA content and original PCA/FA ratio in the forages. The concentration of both phenolic acids in culture fluids was low after a 72 h incubation, and the mixed rumen microorganisms metabolized nearly all phenolic acids released into the culture fluids. FAest digestibility in the whole digestive tract was negatively correlated with dietary PCA/FA ratio, but a converse result occurred with dietary PCAest digestibility. The digestibility in either the rumen or the whole digestive tract was greater for FAest than for PCAest. Forage PCAest in comparison with FAest is not easily digested in either the rumen or the whole digestive tract, and they were negatively affected by forage FAeth content and lignification extent indicated by the original dietary PCA/FA ratio. © 2015 Society of Chemical Industry.

Comparison of the Digestibility of the Major Peanut Allergens in Thermally Processed Peanuts and in Pure Form

PubMed Central

Maleki, Soheila J.; Schmitt, David A.; Galeano, Maria; Hurlburt, Barry K.

2014-01-01

It has been suggested that the boiling or frying of peanuts leads to less allergenic products than roasting. Here, we have compared the digestibility of the major peanut allergens in the context of peanuts subjected to boiling, frying or roasting and in purified form. The soluble peanut extracts and the purified allergens were digested with either trypsin or pepsin and analyzed by gel electrophoresis and western blot. T-cell proliferation was measured for the purified allergens. In most cases, boiled and raw peanut proteins were similarly digestible, but the Ara h 1 protein in the boiled extracts was more resistant to digestion. Most proteins from fried and roasted peanuts were more resistant to digestion than in raw and boiled samples, and more IgE binding fragments survived digestion. High-molecular-weight fragments of Ara h1 were resistant to digestion in fried and roasted samples. Ara h 1 and Ara h 2 purified from roasted peanuts were the most resistant to digestion, but differed in their ability to stimulate T-cells. The differences in digestibility and IgE binding properties of the major allergens in roasted, fried and boiled peanuts may not explain the difference between the prevalence of peanut allergy in different countries that consume peanut following these varied processing methods. PMID:28234320

Life Cycle Assessment of Biogas Production in Small-scale Household Digesters in Vietnam

PubMed Central

Vu, T. K. V.; Vu, D. Q.; Jensen, L. S.; Sommer, S. G.; Bruun, S.

2015-01-01

Small-scale household digesters have been promoted across Asia as a sustainable way of handling manure. The major advantages are that they produce biogas and reduce odor. However their disadvantages include the low recycling of nutrients, because digestate is dilute and therefore difficult to transport, and the loss of biogas as a result of cracks and the intentional release of excess biogas. In this study, life cycle assessment (LCA) methodology was used to assess the environmental impacts associated with biogas digesters in Vietnam. Handling 1,000 kg of liquid manure and 100 kg of solid manure in a system with a biogas digester reduced the impact potential from 4.4 kg carbon dioxide (CO2) equivalents to 3.2 kg CO2 equivalents compared with traditional manure management. However, this advantage could easily be compromised if digester construction is considered in the LCA or in situations where there is an excess of biogas which is intentionally released. A sensitivity analysis showed that biogas digesters could be a means of reducing global warming if methane emissions can be kept low. In terms of eutrophication, farms with biogas digesters had 3 to 4 times greater impacts. In order to make biogas digesters sustainable, methods for recycling digestates are urgently required. PMID:25715690

Activated carbon enhanced anaerobic digestion of food waste - Laboratory-scale and Pilot-scale operation.

PubMed

Zhang, Le; Zhang, Jingxin; Loh, Kai-Chee

2018-05-01

Effects of activated carbon (AC) supplementation on anaerobic digestion (AD) of food waste were elucidated in lab- and pilot-scales. Lab-scale AD was performed in 1 L and 8 L digesters, while pilot-scale AD was conducted in a 1000 L digester. Based on the optimal dose of 15 g AC per working volume derived from the 1 L digester, for the same AC dosage in the 8 L digester, an improved operation stability coupled with a higher methane yield was achieved even when digesters without AC supplementation failed after 59 days due to accumulation of substantial organic intermediates. At the same time, color removal from the liquid phase of the digestate was dramatically enhanced and the particle size of the digestate solids was increased by 53% through AC supplementation after running for 59 days. Pyrosequencing of 16S rRNA gene showed the abundance of predominant phyla Firmicutes, Elusimicrobia and Proteobacteria selectively enhanced by 1.7-fold, 2.9-fold and 2.1-fold, respectively. Pilot-scale digester without AC gave an average methane yield of 0.466 L⋅(gVS) -1 ⋅d -1 at a composition of 53-61% v/v methane. With AC augmentation, an increase of 41% in methane yield was achieved in the 1000 L digester under optimal organic loading rate (1.6 g VS FW ·L -1 ·d -1 ). Copyright © 2018 Elsevier Ltd. All rights reserved.

Anaerobic Digestion Alters Copper and Zinc Speciation.

PubMed

Legros, Samuel; Levard, Clément; Marcato-Romain, Claire-Emmanuelle; Guiresse, Maritxu; Doelsch, Emmanuel

2017-09-19

Anaerobic digestion is a widely used organic waste treatment process. However, little is known on how it could alter the speciation of contaminants in organic waste. This study was focused on determining the influence of anaerobic digestion on the speciation of copper and zinc, two metals that generally occur at high concentration in organic waste. Copper and zinc speciation was investigated by X-ray absorption spectroscopy in four different raw organic wastes (predigestion) and their digested counterparts (postdigestion, i.e., digestates). The results highlighted an increase in the digestates of the proportion of amorphous or nanostructured copper sulfides as well as amorphous or nanostructured zinc sulfides and zinc phosphate as compared to raw waste. We therefore suggest that the environmental fate of these elements would be different when spreading either digestates or raw waste on cropland.

Improved Starch Digestion of Sucrase-deficient Shrews Treated With Oral Glucoamylase Enzyme Supplements.

PubMed

Nichols, Buford L; Avery, Stephen E; Quezada-Calvillo, Roberto; Kilani, Shadi B; Lin, Amy Hui-Mei; Burrin, Douglas G; Hodges, Benjamin E; Chacko, Shaji K; Opekun, Antone R; Hindawy, Marwa El; Hamaker, Bruce R; Oda, Sen-Ichi

2017-08-01

Although named because of its sucrose hydrolytic activity, this mucosal enzyme plays a leading role in starch digestion because of its maltase and glucoamylase activities. Sucrase-deficient mutant shrews, Suncus murinus, were used as a model to investigate starch digestion in patients with congenital sucrase-isomaltase deficiency.Starch digestion is much more complex than sucrose digestion. Six enzyme activities, 2 α-amylases (Amy), and 4 mucosal α-glucosidases (maltases), including maltase-glucoamylase (Mgam) and sucrase-isomaltase (Si) subunit activities, are needed to digest starch to absorbable free glucose. Amy breaks down insoluble starch to soluble dextrins; mucosal Mgam and Si can either directly digest starch to glucose or convert the post-α-amylolytic dextrins to glucose. Starch digestion is reduced because of sucrase deficiency and oral glucoamylase enzyme supplement can correct the starch maldigestion. The aim of the present study was to measure glucogenesis in suc/suc shrews after feeding of starch and improvement of glucogenesis by oral glucoamylase supplements. Sucrase mutant (suc/suc) and heterozygous (+/suc) shrews were fed with C-enriched starch diets. Glucogenesis derived from starch was measured as blood C-glucose enrichment and oral recombinant C-terminal Mgam glucoamylase (M20) was supplemented to improve starch digestion. After feedings, suc/suc and +/suc shrews had different starch digestions as shown by blood glucose enrichment and the suc/suc had lower total glucose concentrations. Oral supplements of glucoamylase increased suc/suc total blood glucose and quantitative starch digestion to glucose. Sucrase deficiency, in this model of congenital sucrase-isomaltase deficiency, reduces blood glucose response to starch feeding. Supplementing the diet with oral recombinant glucoamylase significantly improved starch digestion in the sucrase-deficient shrew.

Small intestinal digestion of raw cornstarch in cattle consuming a soybean hull-based diet is improved by duodenal casein infusion.

PubMed

Brake, D W; Titgemeyer, E C; Bailey, E A; Anderson, D E

2014-09-01

Six duodenally and ileally cannulated steers were used in 3 sequential studies to measure 1) basal nutrient flows from a soybean hull-based diet, 2) small intestinal digestibility of raw cornstarch continuously infused into the duodenum, and 3) responses of small intestinal starch digestion to duodenal infusion of 200 or 400 g/d casein. Our objective was to evaluate responses in small intestinal starch digestion in cattle over time and to measure responses in small intestinal starch digestion to increasing amounts of MP. On average, cattle consumed 3.7 kg/d DM, 68 g/d dietary N, and 70 g/d dietary starch. Starch flow to the duodenum was small (38 g/d), and N flow was 91 g/d. Small intestinal digestibility of duodenal N was 57%, and small intestinal digestion of duodenal starch flow was extensive (92%). Small intestinal starch digestibility was 34% when 1.5 kg/d raw cornstarch was continuously infused into the duodenum. Subsequently, cattle were placed in 1 of 2 replicated Latin squares that were balanced for carryover effects to determine response to casein infusions and time required for adaptation. Duodenal infusion of casein linearly increased (P ≤ 0.05) small intestinal starch digestibility, and small intestinal starch digestion adapted to infusion of casein in 6 d. Ethanol-soluble starch and unpolymerized glucose flowing to the ileum increased linearly (P ≤ 0.05) with increasing infusion of casein. Plasma cholecystokinin was not affected by casein infusion, but circulating levels of glucose were increased by casein supplementation (P ≤ 0.05). Responses in small intestinal starch digestion in cattle adapted to casein within 6 d, and increases in duodenal supply of casein up to 400 g/d increased small intestinal starch digestion in cattle.

Formation of reactive aldehydes (MDA, HHE, HNE) during the digestion of cod liver oil: comparison of human and porcine in vitro digestion models.

PubMed

Tullberg, Cecilia; Larsson, Karin; Carlsson, Nils-Gunnar; Comi, Irene; Scheers, Nathalie; Vegarud, Gerd; Undeland, Ingrid

2016-03-01

In this work, we investigated lipid oxidation of cod liver oil during gastrointestinal (GI) digestion using two types of in vitro digestion models. In the first type of model, we used human GI juices, while we used digestive enzymes and bile from porcine origin in the second type of model. Human and porcine models were matched with respect to factors important for lipolysis, using a standardized digestion protocol. The digests were analysed for reactive oxidation products: malondialdehyde (MDA), 4-hydroxy-trans-2-nonenal (HNE), and 4-hydroxy-trans-2-hexenal (HHE) by liquid chromatography/atmospheric pressure chemical ionization-mass spectrometry (LC/APCI-MS), and for free fatty acids (FFA) obtained during the digestion by gas chromatography-mass spectrometry (GC-MS). The formation of the oxidation products MDA, HHE, and HNE was low during the gastric digestion, however, it increased during the duodenal digestion. The formation of the oxidation products reached higher levels when digestive juices of human origin were used (60 μM of MDA, 0.96 μM of HHE, and 1.6 μM of HNE) compared to when using enzymes and bile of porcine origin (9.8, and 0.36 μM of MDA; 0.16, and 0.026 μM of HHE; 0.23, and 0.005 μM of HNE, respectively, in porcine models I and II). In all models, FFA release was only detected during the intestinal step, and reached up to 31% of total fatty acids (FA). The findings in this work may be of importance when designing oxidation oriented lipid digestion studies.

Digestion of chrysanthemum stunt viroid by leaf extracts of Capsicum chinense indicates strong RNA-digesting activity.

PubMed

Iraklis, Boubourakas; Kanda, Hiroko; Nabeshima, Tomoyuki; Onda, Mayu; Ota, Nao; Koeda, Sota; Hosokawa, Munetaka

2016-08-01

CSVd could not infect Nicotiana benthamiana when the plants were pretreated with crude leaf extract of Capsicum chinense 'Sy-2'. C. chinense leaves were revealed to contain strong RNA-digesting activity. Several studies have identified active antiviral and antiviroid agents in plants. Capsicum plants are known to contain antiviral agents, but the mechanism of their activity has not been determined. We aimed to elucidate the mechanism of Capsicum extract's antiviroid activity. Chrysanthemum stunt viroid (CSVd) was inoculated into Nicotiana benthamiana plants before or after treating the plants with a leaf extract of Capsicum chinense 'Sy-2'. CSVd infection was determined using quantitative reverse transcription-polymerase chain reaction (qRT-PCR) 3 weeks after inoculation. When Capsicum extract was sprayed or painted onto N. benthamiana before inoculation, it was effective in preventing infection by CSVd. To evaluate CSVd digestion activity in leaf extracts, CSVd was mixed with leaf extracts of Mirabilis, Phytolacca, Pelargonium and Capsicum. CSVd-digesting activities were examined by quantifying undigested CSVd using qRT-PCR, and RNA gel blotting permitted visualization of the digested CSVd. Only Capsicum leaf extract digested CSVd, and in the Capsicum treatment, small digested CSVd products were detected by RNA gel blot analysis. When the digesting experiment was performed for various cultivars and species of Capsicum, only cultivars of C. chinense showed strong CSVd-digesting activity. Our observations indicated that Capsicum extract contains strong RNA-digesting activity, leading to the conclusion that this activity is the main mechanism for protection from infection by CSVd through spraying or painting before inoculation. To our knowledge, this is the first report of a strong RNA-digesting activity by a plant extract.

Benefits to decomposition rates when using digestate as compost co-feedstock: Part I - Focus on physicochemical parameters.

PubMed

Arab, Golnaz; McCartney, Daryl

2017-10-01

Anaerobic digestion (AD) has gained a significant role in municipal solid waste management, but managing a high volume of digestate is one of the challenges with AD technology. One option is to mix digestate with fresh and/or stabilized organic waste and then feed to the composting process. In this study, the effect of co-composting anaerobic digestate (in different quantities) on a composting process was investigated. The digestate was prepared in a pilot-scale 500L high solids dry anaerobic digester and composting was completed in eight 25L reactors with different ratios of digestate to fresh feedstock from the organic fraction of municipal solid waste (OFMSW). The digestate constituted 0, 10, 20, 30, 40, 50, 75, or 100% (wet mass) of the feedstock. The co-composting experiment was conducted in two phases: active aeration and curing. Monitored parameters included: process temperature, aeration rate, oxygen concentration of the outlet gas, mass changes, total solids, organic matter, pH, and electrical conductivity. In addition, respirometry, C:N ratio, ammonium to nitrate ratio, and Solvita® tests were used to quantify stability and maturity end points. The results showed that the addition of digestate to the OFMSW increased composting reaction rates in all cases, with peak performance occurring within the ratio of 20-40% of digestate addition on a wet weight basis. Reactor performance may have been influenced by the high total ammonia nitrogen (TAN) levels in the digestate. Composting rates increased as TAN levels increased up to 5000 TAN mgkg -1 DM; however, TAN may have become inhibitory at higher levels. Crown Copyright © 2017. Published by Elsevier Ltd. All rights reserved.

Effects of Aircraft Noise and Sonic Booms on Domestic Animals and Wildlife: A Literature Synthesis

DTIC Science & Technology

1988-06-01

the digestive response of yearling wethers to the same sound types and intensities used in the two studies above: white noise and music presen’ed...metabolizable energy of the ration and improved the apparent nutrient digestibilities . Sound intensity did not affect apparent digest - ibility coefficients. The...high digestibility coefficients for lambs exposed to intermittent sounds suggests that those types of auditory stimuli influenced the digestive system

Relationship between in vitro assays and standardized ileal amino acid digestibility of animal protein meals in broilers.

PubMed

Rochell, S J; Kuhlers, D L; Dozier, W A

2013-01-01

Two identical trials were conducted to determine the relationship of a novel digestive enzyme assay, Poultry Complete IDEA (PC IDEA), and the pepsin digestibility assay with standardized ileal amino acid digestibility (SIAAD) of 20 animal protein meals (APM) fed to broilers from 25 to 30 d of age. Animal protein meals included 10 meat and bone meals (MBM) consisting of bovine, porcine, or mixed bovine and porcine raw materials (BP), and 10 animal protein blends containing animal proteins from various species. Treatments consisted of 20 semi-purified diets containing 1 APM as the sole source of dietary amino acids (AA), and 1 N-free diet to determine endogenous ileal AA flow. With the exception of the N-free diet, diets were formulated to contain 20% CP. In each trial, 756 Ross × Ross 708 male broilers were housed in battery cages and randomly assigned to 21 dietary treatments on d 25 (12 birds per cage; 3 replicate cages), and ileal digesta were collected on d 30 for determination of SIAAD. Pepsin digestibility and PC IDEA were determined for APM samples from each experimental diet (3 replicates per trial; 6 total replicates). Pepsin digestibility and PC IDEA were both correlated (P < 0.001) with SIAAD for each AA. Multiple linear regression of PC IDEA and pepsin digestibility on SIAAD resulted in the following equations: % Lys SIAAD = [-9.65 + (0.38 × % PC IDEA predicted Lys digestibility) + (0.69 × % pepsin digestibility)], % Met SIAAD = [-35.95 + (0.62 × % PC IDEA predicted Met digestibility) + (0.75 × % pepsin digestibility)], % Thr SIAAD = [-77.5 + (0.39 × % PC IDEA predicted Thr digestibility) + (1.37 × % pepsin digestibility)]. Values of R(2) were 0.46, 0.47, and 0.55 for Lys, Met, and Thr, respectively. The relatively low R(2) values may have been due to the limited range in SIAAD observed for the 20 APM, and additional data on APM varying in SIAAD are needed.

Testing low cost anaerobic digestion (AD) systems

USDA-ARS?s Scientific Manuscript database

To evaluate the potential for low technology and low cost digesters for small dairies, BARC and researchers from the University of Maryland installed six modified Taiwanese-model field-scale (FS) digesters near the original dairy manure digester. The FS units receive the same post-separated liquid ...

Inactivation of dairy manure-borne pathogens by anaerobic digestion

USDA-ARS?s Scientific Manuscript database

Background: Anaerobic digestion of animal manure has the potential to inactivate enteric pathogens, thereby reducing exposures to livestock and humans when the products of digestion are disposed by land-spreading or irrigation or returned to livestock uses such as bedding. Data on digester effectiv...

40 CFR Appendix D to Part 136 - Precision and Recovery Statements for Methods for Measuring Metals

Code of Federal Regulations, 2012 CFR

2012-07-01

... water and three effluents. These samples were digested by the total digestion procedure, 4.1.3 in this.... These samples were digested by the total digestion procedure, 4.1.3 in this manual. Results for the...

40 CFR Appendix D to Part 136 - Precision and Recovery Statements for Methods for Measuring Metals

Code of Federal Regulations, 2014 CFR

2014-07-01

... water and three effluents. These samples were digested by the total digestion procedure, 4.1.3 in this.... These samples were digested by the total digestion procedure, 4.1.3 in this manual. Results for the...

40 CFR Appendix D to Part 136 - Precision and Recovery Statements for Methods for Measuring Metals

Code of Federal Regulations, 2013 CFR

2013-07-01

... water and three effluents. These samples were digested by the total digestion procedure, 4.1.3 in this.... These samples were digested by the total digestion procedure, 4.1.3 in this manual. Results for the...

Slowly digestible starch diets alter proximal glucosidase activity and glucose absorption

USDA-ARS?s Scientific Manuscript database

Sucrase-isomaltase (Si) and maltase-glucoamylase (Mgam) are mucosal glucosidases required for digestion of starch to glucose. Ablation of maltase-Mgam reduces in vivo starch digestion. We tested whether slowly digestible starch diets induce changes in glucosidase activities. Rice starch was encaps...

Quantitative measurement of protein digestion in simulated gastric fluid.

PubMed

Herman, Rod A; Korjagin, Valerie A; Schafer, Barry W

2005-04-01

The digestibility of novel proteins in simulated gastric fluid is considered to be an indicator of reduced risk of allergenic potential in food, and estimates of digestibility for transgenic proteins expressed in crops are required for making a human-health risk assessment by regulatory authorities. The estimation of first-order rate constants for digestion under conditions of low substrate concentration was explored for two protein substrates (azocoll and DQ-ovalbumin). Data conformed to first-order kinetics, and half-lives were relatively insensitive to significant variations in both substrate and pepsin concentration when high purity pepsin preparations were used. Estimation of digestion efficiency using densitometric measurements of relative protein concentration based on SDS-PAGE corroborated digestion estimates based on measurements of dye or fluorescence release from the labeled substrates. The suitability of first-order rate constants for estimating the efficiency of the pepsin digestion of novel proteins is discussed. Results further support a kinetic approach as appropriate for comparing the digestibility of proteins in simulated gastric fluid.

Evaluation studies of persimmon plant (Diospyros kaki) for physiological benefits and bioaccessibility of antioxidants by in vitro simulated gastrointestinal digestion.

PubMed

Martínez-Las Heras, Ruth; Pinazo, Alicia; Heredia, Ana; Andrés, Ana

2017-01-01

This study aims to analyze the antioxidant benefits from persimmon leaf tea, fruit and fibres taking into account their changes along gastrointestinal digestion. The evolution of polyphenols, flavonoids and antioxidant capacity was studied using the recent harmonized in vitro protocol published by Minekus et al. (2014). The digestion was performed with and without digestive enzymes. Results showed aqueous leaf extract was richer in antioxidants than the fruit or the extracted fibres. Nevertheless, persimmon-leaf antioxidants were more sensitive to the digestive environment. In general, the oral conditions greatly affected the antioxidants, while gastric digestion led to slight additional losses. The intestinal step enhanced polyphenols and flavonoids solubility coming from the fruit and fibres. Additionally, the presence of digestive enzymes positively contributed to antioxidant release throughout digestion. Finally, the bioaccessibility of polyphenols, flavonoids and antioxidant activity of persimmon fruit were 1.4, 1.0 and 3.8 times higher than in aqueous leaf extract. Copyright © 2016 Elsevier Ltd. All rights reserved.

Heat inactivation of poliovirus in wastewater sludge

DOE Office of Scientific and Technical Information (OSTI.GOV)

Ward, R.L.; Ashley, C.S.; Moseley, R.H.

1976-09-01

The effect of raw and anaerobically digested sludge on heat inactivation of poliovirus was investigated. Raw sludge was found to be very protective of poliovirus plaque-forming ability at all temperatures studied, but digested sludge had variable effects that were highly dependent upon the experimental conditions. In low concentrations and at relatively low inactivation temperatures, digested sludge is nearly as protective of poliovirus as raw sludge. However, at higher temperatures and concentrations, digested sludge caused a significant acceleration of poliovirus inactivation. The difference between the protective capability of raw and digested sludge is not due to loss of protective material, becausemore » this component is present in the solids of digested sludge as well as in those of raw sludge. Instead, the difference is due to a virucidal agent acquired during digestion. Addition of this agent to the solids of either raw or digested sludge reverses the protective potential of these solids during heat treatment of poliovirus.« less

[Anaerobic digestion of animal manure contaminated by tetracyclines].

PubMed

Tong, Zi-Lin; Liu, Yuan-Lu; Hu, Zhen-Hu; Yuan, Shou-Jun

2012-03-01

Anaerobic digestion of pig manure spiked with tetracycline (TC) and chlortetracycline (CTC) and the degradation of the two antibiotics during the anaerobic digestion at 35 degrees C were investigated. The results indicate that propionate was the main volatile fatty acid produced during the anaerobic digestion followed by acetate. Compared with the CTC addition, TC + CTC addition showed obvious inhibitory effect on the hydrolysis and acidification of easily digestible organic components of pig manure. The cumulative methane production of TC, CTC, TC + CTC and CK2 during anaerobic digestion was 386.4 mL, 406.0 mL, 412.1 mL and 464.6 mL, respectively. Degradation of TC and CTC followed the first-order kinetic equation. The half-life of TC and CTC was 14-18 days and 10 days, respectively. After the treatment of 45-day anaerobic digestion, the degradation efficiency of TC was 88.6%-91.6% with 97.7%-98.2% of CTC. Therefore, anaerobic digestion shows the benefit on the management of animal manures contaminated by tetracyclines.

A first step towards a consensus static in vitro model for simulating full-term infant digestion.

PubMed

Ménard, O; Bourlieu, C; De Oliveira, S C; Dellarosa, N; Laghi, L; Carrière, F; Capozzi, F; Dupont, D; Deglaire, A

2018-02-01

In vitro alternatives to clinical trials are used for studying human food digestion. For simulating infant digestion, only a few models, lacking physiological relevance, are available. Thanks to an extensive literature review of the in vivo infant digestive conditions, a gastrointestinal static in vitro model was developed for infants born at term and aged 28days. The model was applied to the digestion of a commercial infant formula. Kinetics of digestion, as well as the structural evolution, were compared with those obtained while submitting the same formula to the adult international consensus protocol of in vitro static digestion. The kinetics of proteolysis and lipolysis differed according to the physiological stage resulting mainly from the reduced level of enzymes and bile salts, as well as the higher gastric pH in the infant model. This in vitro static model of infant digestion is of interest for scientists, food or pharmaceutical manufacturers. Copyright © 2017 Elsevier Ltd. All rights reserved.

Comparative nutrient digestibility of arctic foxes (Alopex lagopus) on Svalbard and farm-raised blue foxes (Alopex lagopus).

PubMed

Ahlstrøm, Øystein; Fuglei, Eva; Mydland, Liv Torunn

2003-01-01

Arctic foxes from Svalbard (n=4) and farmed blue foxes (n=4) was used in a digestibility experiment with a high-carbohydrate feed to add more information to the nutritional physiology of the arctic fox, and to compare its digestive capacity with that of the farmed blue fox. The arctic fox has a diet containing mainly protein and fat from mammals and birds, while farmed blue foxes have been exposed to an omnivorous dietary regime for more than 80 generations. The experiment showed in general no difference in digestive capacity for protein and fat between the foxes (P>0.05), but for carbohydrates, including starch and glucose, the blue fox revealed higher digestibility values. The superior digestive capacity for carbohydrates in blue fox might be a result of a long-term selection of animals digesting dietary carbohydrates more efficiently, or that an early age exposition to dietary carbohydrates has given permanent improvement of the carbohydrate digestion in the gut.

In Vitro Pepsin Digestibility of Cooked Proso Millet ( Pancium miliaceum L.) and Related Species from Around the World.

PubMed

Gulati, Paridhi; Jia, Shangang; Li, Aixia; Holding, David Richard; Santra, Dipak; Rose, Devin Jerrold

2018-06-20

Thirty-three accessions of proso millet (Panicum miliaceum) with different countries of origin were screened for their pepsin digestibility after cooking to identify samples with high digestibility. The pepsin digestibility of all samples ranged from 26 to 57% (average 32%). There were no apparent differences in protein profiles (SDS-PAGE) of samples with the lowest, intermediate, and highest digestibilities. However, LC-MS/MS analysis revealed a negative correlation between pepsin digestibility and peptides that matched to high molecular weight proteins (24 kDa) from Panicum hallii with regions of contiguous hydrophobic amino acids. Low digestibility upon cooking was also observed for other species from the Panicum genus, such as little millet, switchgrass and panicgrass, which suggests a unique inherent property of the genus. The obtained results from this study may form a basis for in-depth analysis of proso proteins that may help in developing new cultivars with higher digestibility upon cooking.

Effect of biopolymer encapsulation on the digestibility of lipid and cholesterol oxidation products in beef during in vitro human digestion.

PubMed

Hur, Sun Jin; Lee, Seung Yuan; Lee, Seung-Jae

2015-01-01

In this study, beef patties were encapsulated with 3% chitosan, pectin, onion powder, or green tea powder and the beef patties were then passed through an in vitro human digestion model. The total lipid digestibility was lowest (p<0.05) in beef patties encapsulated with chitosan and pectin after digestion in the small intestine. Thiobarbituric acid reactive substance (TBARS) values were significantly lower (p<0.05) for beef patties encapsulated with chitosan and pectin, when compared with the control, after digestion in the small intestine. In contrast, the 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) radical-scavenging activity was highest (p<0.05) in beef patties encapsulated with onion powder and green tea powder after digestion in the small intestine. The total cholesterol oxidation product (COP) content was significantly lower (p<0.05) in beef patties encapsulated with biopolymers than in the control after digestion in the small intestine. Copyright © 2014 Elsevier Ltd. All rights reserved.

Impact of different ratios of feedstock to liquid anaerobic digestion effluent on the performance and microbiome of solid-state anaerobic digesters digesting corn stover.

PubMed

Li, Yueh-Fen; Shi, Jian; Nelson, Michael C; Chen, Po-Hsu; Graf, Joerg; Li, Yebo; Yu, Zhongtang

2016-01-01

The objective of this study was to understand how the non-microbial factors of L-AD effluent affected the microbiome composition and successions in the SS-AD digesters using both Illumina sequencing and qPCR quantification of major genera of methanogens. The SS-AD digesters started with a feedstock/total effluent (F/Et) ratio 2.2 (half of the effluent was autoclaved) performed stably, while the SS-AD digesters started with a 4.4 F/Et ratio (no autoclaved effluent) suffered from digester acidification, accumulation of volatile fatty acids, and ceased biogas production two weeks after startup. Some bacteria and methanogens were affected by non-microbial factors of the L-AD fluent. Alkalinity, the main difference between the two F/Et ratios, may be the crucial factor when SS-AD digesters were started using L-AD effluent. Copyright © 2015 Elsevier Ltd. All rights reserved.

Evaluation of anaerobic degradation, biogas and digestate production of cereal silages using nylon-bags.

PubMed

Negri, Marco; Bacenetti, Jacopo; Fiala, Marco; Bocchi, Stefano

2016-06-01

In this study, the degradation efficiency and the biogas and digestate production during anaerobic digestion were evaluated for the cereal silages most used to feed biogas plants. To this purpose, silages of: maize from the whole plant, maize from the ear, triticale and wheat were digested, inside of nylon bags, in laboratory scale digesters, for 75days. Overall, the test involved 288 nylon bags. After 75days of digestion, the maize ear silage shows the highest degradation efficiency (about 98%) while wheat silage the lowest (about 83%). The biogas production ranges from 438 to 852Nm(3)/t of dry matter for wheat and ear maize silage, respectively. For all the cereal silages, the degradation as well as the biogas production are faster at the beginning of the digestion time. Digestate mass, expressed as percentage of the fresh matter, ranges from 38% to 84% for wheat and maize ear silage, respectively. Copyright © 2016 Elsevier Ltd. All rights reserved.

Biogas production generated through continuous digestion of natural and cultivated seaweeds with dairy slurry.

PubMed

Tabassum, Muhammad Rizwan; Wall, David M; Murphy, Jerry D

2016-11-01

The technical feasibility of long term anaerobic mono-digestion of two brown seaweeds, and co-digestion of both seaweeds with dairy slurry was investigated whilst increasing the organic loading rate (OLR). One seaweed was natural (L. digitata); the second seaweed (S. Latissima) was cultivated. Higher proportions of L. digitata in co-digestion (66.6%) allowed the digester to operate more efficiently (OLR of 5kgVSm(-3)d(-1) achieving a specific methane yield (SMY) of 232LCH4kg(-1)VS) as compared to lower proportions (33.3%). Co-digestion of 66.6% cultivated S. latissima, with dairy slurry allowed a higher SMY of 252LCH4kg(-1)VS but at a lower OLR of 4kgVSm(-3)d(-1). Optimum conditions for mono-digestion of both seaweeds were effected at 4kgVSm(-3)d(-1). Chloride concentrations increased to high levels in the digestion of both seaweeds but were not detrimental to operation. Copyright © 2016 Elsevier Ltd. All rights reserved.

Biochemical, Transcriptomic and Proteomic Analyses of Digestion in the Scorpion Tityus serrulatus: Insights into Function and Evolution of Digestion in an Ancient Arthropod

PubMed Central

Fuzita, Felipe J.; Pinkse, Martijn W. H.; Patane, José S. L.; Juliano, Maria A.; Verhaert, Peter D. E. M.; Lopes, Adriana R.

2015-01-01

Scorpions are among the oldest terrestrial arthropods and they have passed through small morphological changes during their evolutionary history on land. They are efficient predators capable of capturing and consuming large preys and due to envenomation these animals can become a human health challenge. Understanding the physiology of scorpions can not only lead to evolutionary insights but also is a crucial step in the development of control strategies. However, the digestive process in scorpions has been scarcely studied. In this work, we describe the combinatory use of next generation sequencing, proteomic analysis and biochemical assays in order to investigate the digestive process in the yellow scorpion Tityus serrulatus, mainly focusing in the initial protein digestion. The transcriptome generated database allowed the quantitative identification by mass spectrometry of different enzymes and proteins involved in digestion. All the results suggested that cysteine cathepsins play an important role in protein digestion. Two digestive cysteine cathepsins were isolated and characterized presenting acidic characteristics (pH optima and stability), zymogen conversion to the mature form after acidic activation and a cross-class inhibition by pepstatin. A more elucidative picture of the molecular mechanism of digestion in a scorpion was proposed based on our results from Tityus serrulatus. The midgut and midgut glands (MMG) are composed by secretory and digestive cells. In fasting animals, the secretory granules are ready for the next predation event, containing enzymes needed for alkaline extra-oral digestion which will compose the digestive fluid, such as trypsins, astacins and chitinase. The digestive vacuoles are filled with an acidic proteolytic cocktail to the intracellular digestion composed by cathepsins L, B, F, D and legumain. Other proteins as lipases, carbohydrases, ctenitoxins and a chitolectin with a perithrophin domain were also detected. Evolutionarily, a large gene duplication of cathepsin L occurred in Arachnida with the sequences from ticks being completely divergent from other arachnids probably due to the particular selective pressures over this group. PMID:25875018

Rheological properties of sewage sludge during enhanced anaerobic digestion with microwave-H2O2 pretreatment.

PubMed

Liu, Jibao; Yu, Dawei; Zhang, Jian; Yang, Min; Wang, Yawei; Wei, Yuansong; Tong, Juan

2016-07-01

The rheological behavior of sludge is of serious concern in anaerobic digestion. This study investigated the rheological properties of sewage sludge during enhanced anaerobic digestion with microwave-H2O2 pretreatment (MW-H2O2). The results showed that MW-H2O2 pretreatment resulted in the improvement of sludge flowability and weakening of its viscoelastic properties. Further positive effects on the rheological properties of digested sludge during anaerobic digestion were observed. The flowability was improved with a low level of apparent viscosity. The decrease of the consistency index and increase of the flow behavior index indicated that the strength of the inner structures and non-Newtonian flow characteristics of digested sludge weakened. Both the storage modulus (G') and loss modulus (G″) decreased, indicating that the viscoelastic behavior became weak. These effects were possibly attributed to the changes of the digested sludge micro-structures, such as extracellular polymeric substances (EPS). This study concluded that anaerobic digestion for treating sewage sludge combined with pretreatment is a more favorable option than single anaerobic digestion from the perspective of rheology. Copyright © 2016 Elsevier Ltd. All rights reserved.

The effects of food components on the digestion of DNA by pepsin.

PubMed

Zhang, Yanfang; Wang, Xingyu; Pan, Xiaoming; Liu, Yu; Wang, Hanqing; Dong, Ping; Liang, Xingguo

2016-11-01

Recently, our study found that naked nucleic acids (NAs) can be digested by pepsin. To better understand the fate of dietary DNA in the digestive tract, in this study we investigated the effects of several food compositions on its digestion. The results showed that protein inhibited the digestion of DNA when the protein:DNA ratio was higher than 80:1 (m/m). DNA found in nucleoprotein (NA), which more closely resembles the state of DNA in food, was as efficiently digested as naked DNA. When the carbohydrate:DNA ratio was 50:1-140:1 (m/m), mono-, di- and polysaccharides did not inhibit DNA digestion. NaCl exhibited an inhibitory effect at 300 mM, whereas divalent cations (Ca(2+ )and Mg(2+)) exerted a much stronger inhibitory effect even at 50 mM. The polycation compounds (e.g. chitosan and spermine) showed a significant inhibitory effect at N/P (NH3(+)/PO4(-)) = 10:1. The close relationship between food composition and DNA digestion suggests that dietary habits and food complexes are important for understanding the in vivo fate of the ingested DNA in the digestive tract.

Angiotensin I-converting enzyme-inhibitory activity of the Norwegian autochthonous cheeses Gamalost and Norvegia after in vitro human gastrointestinal digestion.

PubMed

Qureshi, T M; Vegarud, G E; Abrahamsen, R K; Skeie, S

2013-02-01

The angiotensin I-converting enzyme (ACE) inhibitory activity of Gamalost cheese, its pH 4.6-soluble fraction, and Norvegia cheese was monitored before and after digestion with human gastric and duodenal juices. Both Gamalost and Norvegia cheeses showed an increased ACE-inhibitory activity during gastrointestinal digestion. However, only Norvegia showed pronounced increased activity after duodenal digestion. More peptides were detected in digested Gamalost compared with digested Norvegia. Most of the peptides in Gamalost were derived from β-casein (CN), some originated from α(s1)-CN, and only a very few originated from α(s2)-CN and κ-CN. In general, the number of peptides increased during gastrointestinal digestion, whereas some peptides were further degraded and disappeared; however, surprisingly, a few peptides remained stable. The aromatic amino acids, such as Tyr, Phe, and Trp; the positively charged amino acids (Arg and Lys); and Leu increased after simulated gastrointestinal digestion of Gamalost and Norvegia. After in vitro gastrointestinal digestion, both Gamalost and Norvegia showed high ACE-inhibitory activity, which may contribute in lowering of mild hypertension. Copyright © 2013 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Low-heat, mild alkaline pretreatment of switchgrass for anaerobic digestion.

PubMed

Jin, Guang; Bierma, Tom; Walker, Paul M

2014-01-01

This study examines the effectiveness of alkaline pretreatment under mild heat conditions (100°C or 212°F) on the anaerobic co-digestion of switchgrass. The effects of alkaline concentration, types of alkaline, heating time and rinsing were evaluated. In addition to batch studies, continuous-feed studies were performed in triplicate to identify potential digester operational problems caused by switchgrass co-digestion while accounting for uncertainty due to digester variability. Few studies have examined anaerobic digestion of switchgrass or the effects of mild heating to enhance alkaline pretreatment prior to biomass digestion. Results indicate that pretreatment can significantly enhance digestion of coarse-ground (≤ 0.78 cm particle size) switchgrass. Energy conversion efficiency as high as 63% was observed, and was comparable or superior to fine-grinding as a pretreatment method. The optimal NaOH concentration was found to be 5.5% (wt/wt alkaline/biomass) with a 91.7% moisture level. No evidence of operational problems such as solids build-up, poor mixing, or floating materials were observed. These results suggest the use of waste heat from a generator could reduce the concentration of alkaline required to adequately pretreat lignocellulosic feedstock prior to anaerobic digestion.

Extracellular digestion during hyposaline exposure in the Dungeness crab, Cancer magister, and the blue crab, Callinectes sapidus.

PubMed

Curtis, Daniel L; van Breukelen, Frank; McGaw, Iain J

2013-12-01

Extracellular digestive processes were examined in the Dungeness crab, Cancer magister and the blue crab, Callinectes sapidus, during hyposaline exposure. Both species are found in estuaries as adults, but vary in their ability to balance the cardiovascular and respiratory demands of concurrent osmoregulation and digestion. The weak osmoregulator, C. magister, is unable to balance the demands of osmoregulation and digestion. Concordant with observed decreases in oxygen consumption and mechanical digestion, proteolytic digestion within the foregut and hepatopancreas was delayed, resulting in a relative reduction of circulating amino acids post-feeding in low salinity. In contrast, the efficient osmoregulator, C. sapidus, balances the demands of osmoregulation and digestion, and mechanical digestion continues unabated in low salinity. Protease activity in the gut fluid and hepatopancreas showed either no change or a reduction over time. The transport of amino acids into the cells post-feeding is opposed by an efflux of amino acids at the cellular level, and resulted in a build up of amino acids in the hemolymph. Despite differences in the extracellular responses to low salinity exposure following feeding, both species were able to maintain high digestive efficiencies. © 2013.

1H NMR and SPME-GC/MS study of hydrolysis, oxidation and other reactions occurring during in vitro digestion of non-oxidized and oxidized sunflower oil. Formation of hydroxy-octadecadienoates.

PubMed

Nieva-Echevarría, Bárbara; Goicoechea, Encarnación; Manzanos, María J; Guillén, María D

2017-01-01

Both fresh and slightly oxidized sunflower oils, as models of omega-6 rich lipids, were submitted to in vitro gastrointestinal digestion and studied by 1 H NMR and SPME-GC/MS. Changes in lipolysis degree, lipid composition and oxidative level were studied by 1 H NMR. Three quantitative approaches were used and several equations were newly developed. In oxidized oil digestates slightly lower hydrolysis and a higher advance of oxidation took place during digestion. This latter was evidenced by a greater decrease of lipid unsaturation degree and enhanced generation of oxidation products (cis,trans-hydroperoxy-octadecadienoates, cis,trans- and trans,trans-hydroxy-octadecadienoates). For the first time, the generation of hydroxy-octadecadienoates during in vitro digestion is reported. Furthermore, SPME-GC/MS study of non-digested and digested samples headspaces confirmed that lipid oxidation occurred: abundances of volatile markers increased (including potentially toxic alpha,beta-unsaturated aldehydes), especially in oxidized oils digestates. Markers of Maillard-type and esterification reactions were also detected in the digestates. Copyright © 2016 Elsevier Ltd. All rights reserved.

Functional Utrastructure of Genlisea (Lentibulariaceae) Digestive Hairs

PubMed Central

Płachno, Bartosz Jan; Kozieradzka-Kiszkurno, Małgorzata; Świątek, Piotr

2007-01-01

Background and Aims Digestive structures of carnivorous plants produce external digestive enzymes, and play the main role in absorption. In Lentibulariaceae, the ultrastructure of digestive hairs has been examined in some detail in Pinguicula and Utricularia, but the sessile digestive hairs of Genlisea have received very little attention so far. The aim of this study was to fill this gap by expanding their morphological, anatomical and histochemical characterization. Methods Several imaging techniques were used, including light, confocal and electron microscopy, to reveal the structure and function of the secretory hairs of Genlisea traps. This report demonstrates the application of cryo-SEM for fast imaging of whole, physically fixed plant secretory structures. Key Results and Conclusion The concentration of digestive hairs along vascular bundles in subgenus Genlisea is a primitive feature, indicating its basal position within the genus. Digestive hairs of Genlisea consist of three compartments with different ultrastructure and function. In subgenus Tayloria the terminal hair cells are transfer cells, but not in species of subgenus Genlisea. A digestive pool of viscous fluid occurs in Genlisea traps. In spite of their similar architecture, the digestive-absorptive hairs of Lentibulariaceae feature differences in morphology and ultrastructure. PMID:17550910

Effect of dosing time on the ammonium nitrogen disinhibition in autothermal thermophilic aerobic digestion for sewage sludge by chemical precipitation.

PubMed

Xu, Changwen; Yuan, Haiping; Lou, Ziyang; Zhang, Guofang; Gong, Junzhe; Zhu, Nanwen

2013-12-01

Magnesium ammonium phosphate (MAP) precipitation was introduced to remove ammonium nitrogen (NH4(+)-N) in autothermal thermophilic aerobic digestion (ATAD) in this study. The dosing time of MgCl2 · 6H2O and NaH2PO4 · 2H2O will influence the removal efficiency of ammonium nitrogen greatly, and the time interval of 2nd, 7th, 12th day were chosen in ATAD process. The lowest NH4(+)-N concentration was found in the 2nd day dosing digester, and 38.37% of VS removal rate was obtained after 12 days digestion, which achieved stabilization 9 days earlier than the non-dosing digester. It revealed that removal of ammonium nitrogen could accelerate the sludge stabilization process. Meanwhile, 49.30% of VS removal rate was found in the 2nd day dosing digester in the 21st day, much higher than that in the non-dosing digester, the 7th day dosing digester, and the 12th day dosing digester, with the corresponding value of 38.37%, 38.38% and 37.04%, respectively. Copyright © 2013 Elsevier Ltd. All rights reserved.

Combined mesophilic anaerobic and thermophilic aerobic digestion process: effect on sludge degradation and variation of sludge property.

PubMed

Cheng, Jiehong; Ji, Yuehong; Kong, Feng; Chen, Xian

2013-12-01

One-stage autothermal thermophilic aerobic digestion (ATAD) is effective for the reduction of volatile solids (VSs) and pathogen in sewage sludges. A novel process of combining mesophilic (<35 °C) anaerobic digestion with a thermophilic (55 °C) aerobic digestion process (AN/TAD) occurred in a one-stage digester, which was designed for aeration energy savings. The efficiency of sludge degradation and variation of sludge properties by batch experiments were evaluated for the AN/TAD digester with an effective volume of 23 L for 30 days compared with conventional thermophilic aerobic digestion (TAD). The AN/TAD system can efficiently achieve sludge stabilization on the 16th day with a VS removal rate of 38.1 %. The AN/TAD system was operated at lower ORP values in a digestion period with higher contents of total organic compounds, volatile fatty acids, protein, and polysaccharide in the soluble phase than those of the TAD system, which can rapidly decreased and had low values in the late period of digestion for the AN/TAD system. In the AN/TAD system, intracellular substances had lysis because of initial hydrolytic acidification.

Frogs as integrative models for understanding digestive organ development and evolution

PubMed Central

Womble, Mandy; Pickett, Melissa; Nascone-Yoder, Nanette

2016-01-01

The digestive system comprises numerous cells, tissues and organs that are essential for the proper assimilation of nutrients and energy. Many aspects of digestive organ function are highly conserved among vertebrates, yet the final anatomical configuration of the gut varies widely between species, especially those with different diets. Improved understanding of the complex molecular and cellular events that orchestrate digestive organ development is pertinent to many areas of biology and medicine, including the regeneration or replacement of diseased organs, the etiology of digestive organ birth defects, and the evolution of specialized features of digestive anatomy. In this review, we highlight specific examples of how investigations using Xenopus laevis frog embryos have revealed insight into the molecular and cellular dynamics of digestive organ patterning and morphogenesis that would have been difficult to obtain in other animal models. Additionally, we discuss recent studies of gut development in non-model frog species with unique feeding strategies, such as Lepidobatrachus laev is and Eleutherodactylouscoqui, which are beginning to provide glimpses of the evolutionary mechanisms that may generate morphological variation in the digestive tract. The unparalleled experimental versatility of frog embryos make them excellent, integrative models for studying digestive organ development across multiple disciplines. PMID:26851628

Cytochemical investigation of the digestive gland of two strombidae species (Strombus gigas and Strombus pugilis) in relation to the nutrition.

PubMed

Volland, Jean-Marie; Gros, Olivier

2012-10-01

Strombus gigas and Strombus pugilis are threatened species and aquaculture represents a good alternative solution to the fishing. In this study, we highlighted the intracellular digestion process in the digestive gland of two Strombidae species, S. gigas and Strombuspugilis, by the cytochemical characterization of two lysosomal enzymes: acid phosphatase and arylsulfatase. In order to check the efficiency of artificial food digestion, we conducted the characterization on freshly collected, starved and artificially fed individuals of S. pugilis. TEM observations of digestive gland sections from freshly collected individuals of both species revealed the presence of acid phosphatase and arylsulfatase activity mostly located in the apical third of digestive cells. Both enzymes were also detected in artificially fed individuals. In response to the starvation, acid phosphatase is not produced anymore by digestive cells, while arylsulfatase is still present. To our knowledge, this is the first cytochemical validation of intracellular digestion of artificial food in Strombidae. This study highlights the intracellular digestion of artificial food developed for Strombidae aquaculture. Moreover, we have shown that the lysosomal activity could be used as a feed index. Copyright © 2012 Wiley Periodicals, Inc.

Effects of inoculum to substrate ratio and co-digestion with bagasse on biogas production of fish waste.

PubMed

Xu, Jie; Mustafa, Ahmed M; Sheng, Kuichuan

2017-10-01

To overcome the biogas inhibition in anaerobic digestion of fish waste (FW), effects of inoculum to substrate ratio (I/S, based on VS) and co-digestion with bagasse on biogas production of FW were studied in batch reactors. I/S value was from 0.95 to 2.55, bagasse content in co-digestion (based on VS) was 25%, 50% and 75%. The highest biogas yield (433.4 mL/gVS) with 73.34% methane content was obtained at an I/S value of 2.19 in mono-digestion of FW; the biogas production was inhibited and the methane content was below 70% when I/S was below 1.5. Co-digestion of FW and bagasse could improve the stability and biogas potential, also reducing the time required to obtain 70% of the total biogas production, although the total biogas yield and methane content decreased with the increase in bagasse content in co-digestion. Biogas yield of 409.5 mL/gVS was obtained in co-digestion of 75% FW and 25% bagasse; simultaneously 78.46% of the total biogas production was achieved after 10 days of digestion.

Anaerobic co-digestion of livestock and vegetable processing wastes: fibre degradation and digestate stability.

PubMed

Molinuevo-Salces, Beatriz; Gómez, Xiomar; Morán, Antonio; García-González, Mari Cruz

2013-06-01

Anaerobic digestion of livestock wastes (swine manure (SM) and poultry litter (PL)) and vegetable processing wastes (VPW) mixtures was evaluated in terms of methane yield, volatile solids removal and lignocellulosic material degradation. Batch experiments were performed with 2% VS (volatile solids) to ensure complete conversion of TVFAs (total volatile fatty acids) and to avoid ammonia inhibition. Experimental methane yields obtained for the mixtures resulted in higher values than those obtained from the sum of the methane yields from the individual components. VPW addition to livestock wastes before anaerobic digestion also resulted in improved VS elimination. In SM-VPW co-digestions, CH4 yield increased from 111 to 244 mL CH4 g VS added(-1), and the percentage of VS removed increased from 50% to 86%. For PL-VPW co-digestions, the corresponding values were increased from 158 to 223 mL CH4 g VS added(-1) and from 70% to 92% VS removed. Hemicelluloses and more than 50% of cellulose were degraded during anaerobic digestion. Thermal analyses indicated that the stabilization of the wastes during anaerobic digestion resulted in significantly less energy being released by digestate samples than fresh samples. Copyright © 2013 Elsevier Ltd. All rights reserved.

The fate and effect of monensin during anaerobic digestion of dairy manure under mesophilic conditions.

PubMed

Arikan, Osman A; Mulbry, Walter; Rice, Clifford; Lansing, Stephanie

2018-01-01

There is growing concern about residual antibiotics and feed additives in the manure of treated animals because of the effects of these residues in the environment. Monensin is the most widely used ionophore coccidiostat in the U.S. The objective of this study was to determine the fate and effect of monensin during the anaerobic digestion of dairy manure. Duplicate plug flow field-scale digesters were operated using non-amended dairy manure and dairy manure amended with monensin to 1 and 10 mg/L for 56 days at 30°C at an organic loading rate of 1.4 kg VS/m3-d and 17-day hydraulic retention time. Results showed that monensin was reduced approximately 70% during anaerobic digestion. Methane production from digesters using manure amended with 1 mg/L monensin was comparable to that from digesters operated without added monensin. However, digesters using manure amended with 10 mg/L monensin yielded 75% less methane than digesters using manure without added monensin. These results suggest that anaerobic digestion is an effective treatment for reducing, but not eliminating, monensin in dairy manure. Monensin did not reduce methane production at concentrations expected in dairy manure at recommended dosage rates.

In vitro digestion of purified β-casein variants A(1), A(2), B, and I: effects on antioxidant and angiotensin-converting enzyme inhibitory capacity.

PubMed

Petrat-Melin, B; Andersen, P; Rasmussen, J T; Poulsen, N A; Larsen, L B; Young, J F

2015-01-01

Genetic polymorphisms of bovine milk proteins affect the protein profile of the milk and, hence, certain technological properties, such as casein (CN) number and cheese yield. However, reports show that such polymorphisms may also affect the health-related properties of milk. Therefore, to gain insight into their digestion pattern and bioactive potential, β-CN was purified from bovine milk originating from cows homozygous for the variants A(1), A(2), B, and I by a combination of cold storage, ultracentrifugation, and acid precipitation. The purity of the isolated β-CN was determined by HPLC, variants were verified by mass spectrometry, and molar extinction coefficients at λ=280nm were determined. β-Casein from each of the variants was subjected to in vitro digestion using pepsin and pancreatic enzymes. Antioxidant and angiotensin-converting enzyme (ACE) inhibitory capacities of the hydrolysates were assessed at 3 stages of digestion and related to that of the undigested samples. Neither molar extinction coefficients nor overall digestibility varied significantly between these 4 variants; however, clear differences in digestion pattern were indicated by gel electrophoresis. In particular, after 60min of pepsin followed by 5min of pancreatic enzyme digestion, one ≈4kDa peptide with the N-terminal sequence (106)H-K-E-M-P-F-P-K- was absent from β-CN variant B. This is likely a result of the (122)Ser to (122)Arg substitution in variant B introducing a novel trypsin cleavage site, leading to the changed digestion pattern. All investigated β-CN variants exhibited a significant increase in antioxidant capacity upon digestion, as measured by the Trolox-equivalent antioxidant capacity assay. After 60min of pepsin + 120min of pancreatic enzyme digestion, the accumulated increase in antioxidant capacity was ≈1.7-fold for the 4 β-CN variants. The ACE inhibitory capacity was also significantly increased by digestion, with the B variant reaching the highest inhibitory capacity at the end of digestion (60min of pepsin + 120min of pancreatic enzymes), possibly because of the observed alternative digestion pattern. These results demonstrate that genetic polymorphisms affect the digestion pattern and bioactivity of milk proteins. Moreover, their capacity for radical scavenging and ACE inhibition is affected by digestion. Copyright © 2015 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Effect of heat and homogenization on in vitro digestion of milk.

PubMed

Tunick, Michael H; Ren, Daxi X; Van Hekken, Diane L; Bonnaillie, Laetitia; Paul, Moushumi; Kwoczak, Raymond; Tomasula, Peggy M

2016-06-01

Central to commercial fluid milk processing is the use of high temperature, short time (HTST) pasteurization to ensure the safety and quality of milk, and homogenization to prevent creaming of fat-containing milk. Ultra-high-temperature sterilization is also applied to milk and is typically used to extend the shelf life of refrigerated, specialty milk products or to provide shelf-stable milk. The structures of the milk proteins and lipids are affected by processing but little information is available on the effects of the individual processes or sequences of processes on digestibility. In this study, raw whole milk was subjected to homogenization, HTST pasteurization, and homogenization followed by HTST or UHT processing. Raw skim milk was subjected to the same heating regimens. In vitro gastrointestinal digestion using a fasting model was then used to detect the processing-induced changes in the proteins and lipids. Using sodium dodecyl sulfate-PAGE, gastric pepsin digestion of the milk samples showed rapid elimination of the casein and α-lactalbumin bands, persistence of the β-lactoglobulin bands, and appearance of casein and whey peptide bands. The bands for β-lactoglobulin were eliminated within the first 15min of intestinal pancreatin digestion. The remaining proteins and peptides of raw, HTST, and UHT skim samples were digested rapidly within the first 15min of intestinal digestion, but intestinal digestion of raw and HTST pasteurized whole milk showed some persistence of the peptides throughout digestion. The availability of more lipid droplets upon homogenization, with greater surface area available for interaction with the peptides, led to persistence of the smaller peptide bands and thus slower intestinal digestion when followed by HTST pasteurization but not by UHT processing, in which the denatured proteins may be more accessible to the digestive enzymes. Homogenization and heat processing also affected the ζ-potential and free fatty acid release during intestinal digestion. Stearic and oleic acids were broken down faster than other fatty acids due to their positions on the outside of the triglyceride molecule. Five different casein phosphopeptide sequences were observed after gastric digestion, and 31 sequences were found after intestinal digestion, with activities yet to be explored. Processing affects milk structure and thus digestion and is an important factor to consider in design of foods that affect health and nutrition. Copyright © 2016 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

A meta-analysis of feed digestion in dairy cows. 1. The effects of forage and concentrate factors on total diet digestibility.

PubMed

Nousiainen, J; Rinne, M; Huhtanen, P

2009-10-01

A meta-analysis based on published experiments with lactating dairy cows was conducted to study the effects of dietary forage and concentrate factors on apparent total diet digestibility. A data set was collected that included a total of 497 dietary treatment means from 92 studies. The diets were based on grass silage or on legume or whole-crop cereal silages partly or completely substituted for grass silage. The silages were supplemented with concentrates given at a flat rate within a dietary comparison. For the statistical evaluation, the data were divided into 5 subsets to quantify silage (digestibility, 42 diets in 17 studies; fermentation characteristics, 108 diets in 39 studies) and concentrate (amount of supplementation, 142 diets in 59 studies; concentration of crude protein, 215 diets in 82 studies; carbohydrate composition, 66 diets in 23 studies) factors on total diet digestibility. The diet digestibility of dairy cows was determined by total fecal collection or by using acid-insoluble ash as an internal marker. Diet organic matter digestibility (OMD) at a maintenance level of feeding (OMD(m)) was estimated using sheep in vivo or corresponding in vitro digestibility values for the forage and reported ingredient and chemical composition values, with tabulated digestibility coefficients for the concentrate components of the diet. A mixed model regression analysis was used to detect the responses of different dietary factors on apparent total diet digestibility. Improved silage OMD(m) resulting from earlier harvest was translated into improved production-level OMD in cows (OMD(p)). The effects of silage fermentation characteristics on OMD(p) were quantitatively small, although sometimes significant. Concentrate supplementation improved total diet OMD(m), but this was not realized in lactating dairy cows because of linearly decreased neutral detergent fiber (NDF) digestibility as concentrate intake increased. Increasing the concentrate crude protein amount quadratically improved OMD(p) in cows, with the response being mostly due to improved NDF digestibility. Replacement of starchy concentrates with fibrous by-products slightly decreased OMD(p) but tended to improve NDF digestibility. The true digestibility of cell solubles (OM - NDF) estimated by the Lucas test both from all data and from the data subsets was not significantly different from 1.00, suggesting that responses in OMD(p) of dairy cows are mediated through changes in the concentration and digestibility of NDF.

Hot topic: apparent total-tract nutrient digestibilities measured commercially using 120-hour in vitro indigestible neutral detergent fiber as a marker are related to commercial dairy cattle performance.

PubMed

Schalla, A; Meyer, L; Meyer, Z; Onetti, S; Schultz, A; Goeser, J

2012-09-01

Measuring individual feed nutrient concentration is common practice for field dairy nutritionists. However, accurately measuring nutrient digestibility and using digestion values in total digestible nutrients models is more challenging. Our objective was to determine if in vivo apparent total-tract nutrient digestibility measured with a practical approach was related to commercial milk production parameters. Total mixed ration and fecal samples were collected from high-producing cows in pens on 39 commercial dairies and analyzed at a commercial feed and forage testing laboratory for nutrient concentration and 120-h indigestible NDF (iNDF) content using the Combs-Goeser in vitro digestion technique. The 120-h iNDF was used as an internal marker to calculate in vivo apparent nutrient digestibilities. Two samples were taken from each dairy and were separated in time by at least 3 wk. Samples were targeted to be taken within 7d of Dairy Herd Improvement (DHI) herd testing. Approved DHI testers measured individual cow milk weights as well as fat and protein concentrations. Individual cow records were averaged by pen corresponding to the total mixed ration and fecal samples. Formulated diet and dry matter intake (DMI) records for each respective pen were also collected. Mixed model regression analysis with dairy specified as a random effect was used to relate explanatory variables (diet nutrient concentrations, formulated DMI, in vivo apparent nutrient digestibilities, and fecal nutrient concentrations) to milk production measures. Dry matter intake, organic matter (OM) digestibility, fecal crude protein (CP) concentration, and fecal ether extract concentration were related to milk, energy-corrected milk, and fat yields. Milk protein concentration was related to CP digestibility, and milk protein yield was related to DMI, OM digestibility, CP digestibility, and ether extract digestibility. Although many studies have related DMI and OM digestibility to milk production under controlled experimental settings, very few have related practical in vivo measures to milk production. By documenting the practical OM digestibility relationship with milk production, nutritionists and scientists may have confidence in this approach for measuring diet performance and collecting nutritional data for commercial dairies. Copyright © 2012 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

In vitro digestibility of individual amino acids in rumen-undegraded protein: the modified three-step procedure and the immobilized digestive enzyme assay.

PubMed

Boucher, S E; Calsamiglia, S; Parsons, C M; Stern, M D; Moreno, M Ruiz; Vázquez-Añón, M; Schwab, C G

2009-08-01

Three soybean meal, 3 SoyPlus (West Central Cooperative, Ralston, IA), 5 distillers dried grains with solubles, and 5 fish meal samples were used to evaluate the modified 3-step in vitro procedure (TSP) and the in vitro immobilized digestive enzyme assay (IDEA; Novus International Inc., St. Louis, MO) for estimating digestibility of AA in rumen-undegraded protein (RUP-AA). In a previous experiment, each sample was ruminally incubated in situ for 16 h, and in vivo digestibility of AA in the intact samples and in the rumen-undegraded residues (RUR) was obtained for all samples using the precision-fed cecectomized rooster assay. For the modified TSP, 5 g of RUR was weighed into polyester bags, which were then heat-sealed and placed into Daisy(II) incubator bottles. Samples were incubated in a pepsin/HCl solution followed by incubation in a pancreatin solution. After this incubation, residues remaining in the bags were analyzed for AA, and digestibility of RUP-AA was calculated based on disappearance from the bags. In vitro RUP-AA digestibility estimates obtained with this procedure were highly correlated to in vivo estimates. Corresponding intact feeds were also analyzed via the pepsin/pancreatin steps of the modified TSP. In vitro estimates of AA digestibility of the feeds were highly correlated to in vivo RUP-AA digestibility, which suggests that the feeds may not need to be ruminally incubated before determining RUP-AA digestibility in vitro. The RUR were also analyzed via the IDEA kits. The IDEA values of the RUR were good predictors of RUP-AA digestibility in soybean meal, SoyPlus, and distillers dried grains with solubles, but the IDEA values were not as good predictors of RUP-AA digestibility in fish meal. However, the IDEA values of intact feed samples were also determined and were highly correlated to in vivo RUP-AA digestibility for all feed types, suggesting that the IDEA value of intact feeds may be a better predictor of RUP-AA digestibility than the IDEA value of the RUR. In conclusion, the modified TSP and IDEA kits are good approaches for estimating RUP-AA digestibility in soybean meal products, distillers dried grains with solubles, and fish meal samples.

40 CFR 428.80 - Applicability; description of the wet digestion reclaimed rubber subcategory.

Code of Federal Regulations, 2014 CFR

2014-07-01

... digestion reclaimed rubber subcategory. 428.80 Section 428.80 Protection of Environment ENVIRONMENTAL... CATEGORY Wet Digestion Reclaimed Rubber Subcategory § 428.80 Applicability; description of the wet digestion reclaimed rubber subcategory. The provisions of this subpart are applicable to process waste water...

40 CFR 428.80 - Applicability; description of the wet digestion reclaimed rubber subcategory.

Code of Federal Regulations, 2012 CFR

2012-07-01

... digestion reclaimed rubber subcategory. 428.80 Section 428.80 Protection of Environment ENVIRONMENTAL... CATEGORY Wet Digestion Reclaimed Rubber Subcategory § 428.80 Applicability; description of the wet digestion reclaimed rubber subcategory. The provisions of this subpart are applicable to process waste water...

40 CFR 428.80 - Applicability; description of the wet digestion reclaimed rubber subcategory.

Code of Federal Regulations, 2013 CFR

2013-07-01

... digestion reclaimed rubber subcategory. 428.80 Section 428.80 Protection of Environment ENVIRONMENTAL... CATEGORY Wet Digestion Reclaimed Rubber Subcategory § 428.80 Applicability; description of the wet digestion reclaimed rubber subcategory. The provisions of this subpart are applicable to process waste water...

Apparent seed digestibility and germination of seeds after passage through the digestive system of northern bobwhite

USDA-ARS?s Scientific Manuscript database

Limited information is available regarding the digestibility or germination of seed after the passage through the digestive system of northern bobwhites (Colinus virginianus), especially of plants associated with the sand sagebrush (Artemisia filifolia)-mixed prairie community. Thus, our objectives...

Effect of partitioning the nonfiber carbohydrate fraction and neutral detergent fiber method on digestibility of carbohydrates by dairy cows.

PubMed

Tebbe, A W; Faulkner, M J; Weiss, W P

2017-08-01

Many nutrition models rely on summative equations to estimate feed and diet energy concentrations. These models partition feed into nutrient fractions and multiply the fractions by their estimated true digestibility, and the digestible mass provided by each fraction is then summed and converted to an energy value. Nonfiber carbohydrate (NFC) is used in many models. Although it behaves as a nutritionally uniform fraction, it is a heterogeneous mixture of components. To reduce the heterogeneity, we partitioned NFC into starch and residual organic matter (ROM), which is calculated as 100 - CP - LCFA - ash - starch - NDF, where crude protein (CP), long-chain fatty acids (LCFA), ash, starch, and neutral detergent fiber (NDF) are a percentage of DM. However, the true digestibility of ROM is unknown, and because NDF is contaminated with both ash and CP, those components are subtracted twice. The effect of ash and CP contamination of NDF on in vivo digestibility of NDF and ROM was evaluated using data from 2 total-collection digestibility experiments using lactating dairy cows. Digestibility of NDF was greater when it was corrected for ash and CP than without correction. Conversely, ROM apparent digestibility decreased when NDF was corrected for contamination. Although correcting for contamination statistically increased NDF digestibility, the effect was small; the average increase was 3.4%. The decrease in ROM digestibility was 7.4%. True digestibility of ROM is needed to incorporate ROM into summative equations. Data from multiple digestibility experiments (38 diets) using dairy cows were collated, and ROM concentrations were regressed on concentration of digestible ROM (ROM was calculated without adjusting for ash and CP contamination). The estimated true digestibility coefficient of ROM was 0.96 (SE = 0.021), and metabolic fecal ROM was 3.43 g/100 g of dry matter intake (SE = 0.30). Using a smaller data set (7 diets), estimated true digestibility of ROM when calculated using NDF corrected for ash and CP contamination was 0.87 (SE = 0.025), and metabolic fecal ROM was 3.76 g/100 g (SE = 0.60). Regardless of NDF method, ROM exhibited nutritional uniformity. The ROM fraction also had lower errors associated with the estimated true digestibility and its metabolic fecal fraction than did NFC. Therefore, ROM may result in more accurate estimates of available energy if integrated into models. Copyright © 2017 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Assessment of by-products of bioenergy systems (anaerobic digestion and gasification) as potential crop nutrient.

PubMed

Kataki, Sampriti; Hazarika, Samarendra; Baruah, D C

2017-01-01

Alternative fertilizer resources have drawn attention in recent times in order to cope up with ever increasing demand for fertilizer. By-products of bioenergy system are considered favourable as organic fertilizer due to their ability to recycle plant nutrients. Present study evaluates fertilizer suitability of by-products of two bioenergy systems viz. 3 types of anaerobic digestion by-products (digestate) from local surplus biomass such as cowdung, Ipomoea carnea:cowdung (60:40) and ricestraw:green gram stover:cowdung (30:30:40) and one gasification by-product (biochar) from rice husk. Digestates were assessed considering 4 different application options of each viz. whole, solid, liquid and ash from solid digestates. Digestate characteristics (organic matter, macronutrients, micronutrients and heavy metal content) were found to be a function of feedstock and processing (solid liquid separation and ashing). Ipomoea carnea based digestates in all application options showed comparatively higher N, P, K, NH 4 + -N, Ca, Mg, S and micro nutrient content than other digestates. Separation concentrated plant nutrients and organic matter in solid digestates, making these suitable both as organic amendments and fertilizer. Separated liquid digestate shared larger fraction of ammonium nitrogen (61-91% of total content), indicating their suitability as readily available N source. However, fertilizer application of liquid digestate may not match crop requirements due to lower total nutrient concentration. Higher electrical conductivity of the liquid digestates (3.4-9.3mScm -1 ) than solid digestates (1.5-2mScm -1 ) may impart phyto-toxic effect upon fertilization due to salinity. In case of by-products with unstable organic fraction i.e. whole and solid digestates of rice straw:green gram stover:cowdung digestates (Humification index 0.7), further processing (stabilization, composting) may be required to maximize their fertilizer benefit. Heavy metal contents of the by-products were found to be within the permitted range specified for organic fertilizer (vermicompost) in India. However, higher Al content of the digestates in whole, solid and ash phase (0.06-16.97gkg -1 fresh matter) can be a concern in acid soil which may cause Al toxicity. Understanding on agrochemical characteristics of bioenergy by-products with varying feedstock and application option is expected to promote their valorization opportunities considering user specific requirements. In the context of agriculturally dominant but energy deficient rural Indian scenario, integrated production of bioenergy and by-product based fertilizer could be very significant to meet the critical additional requirement of both energy and fertilizer. Copyright © 2016 Elsevier Ltd. All rights reserved.

Effects of the Application of Digestates from Wet and Dry Anaerobic Fermentation to Japanese Paddy and Upland Soils on Short-Term Nitrification

PubMed Central

Sawada, Kozue; Toyota, Koki

2015-01-01

Wet and dry anaerobic fermentation processes are operated for biogas production from organic matter, resulting in wet and dry digestates as by-products, respectively. The application of these digestates to soil as fertilizer has increased in recent years. Therefore, we herein compared the effects of applying wet digestates (pH 8.2, C/N ratio 4.5), dry digestates (pH 8.8, C/N ratio 23.4), and a chemical fertilizer to Japanese paddy and upland soils on short-term nitrification under laboratory aerobic conditions. Chloroform-labile C, an indicator of microbial biomass, was only minimally affected by these applications, indicating that a small amount of labile N was immobilized by microbes. All applications led to rapid increases in NO3 -N contents in both soils, and ammonia-oxidizing bacteria, but not archaea may play a critical role in net nitrification in the amended soils. The net nitrification rates for both soils were the highest after the application of dry digestates, followed by wet digestates and then the chemical fertilizer in order of decreasing soil pH. These results suggest that the immediate effects of applying digestates, especially dry digestates with the highest pH, on nitrate leaching need to be considered when digestates are used as alternative fertilizers. PMID:25740173

Batch anaerobic co-digestion of waste activated sludge and microalgae (Chlorella sorokiniana) at mesophilic temperature.

PubMed

Beltrán, Carolina; Jeison, David; Fermoso, Fernando G; Borja, Rafael

2016-08-23

The microalgae Chlorella sorokiniana are used as co-substrate for waste activated sludge (WAS) anaerobic digestion. The specific objective of this research was to evaluate the feasibility of improving methane production from anaerobic digestion of WAS in co-digestion with this microalga, based on an optimized mixture percentage. Thus, the anaerobic co-digestion of both substrates aims to overcome the drawbacks of the anaerobic digestion of single WAS, simultaneously improving its management. Different co-digestion mixtures (0% WAS-100% microalgae; 25% WAS-75% microalgae; 50% WAS-50% microalgae; 75% WAS-25% microalgae; 100% WAS-0% microalgae) were studied. The highest methane yield (442 mL CH4/g VS) was obtained for the mixture with 75% WAS and 25% microalgae. This value was 22% and 39% higher than that obtained in the anaerobic digestion of the sole substrates WAS and microalgae, respectively, as well as 16% and 25% higher than those obtained for the co-digestion mixtures with 25% WAS and 75% microalgae and 50% WAS and 50% microalgae, respectively. The kinetic constant of the process increased 42%, 42% and 12%, respectively, for the mixtures with 25%, 50% and 75% of WAS compared to the substrate without WAS. Anaerobic digestion of WAS, together with C. sorokiniana, has been clearly improved by ensuring its viability, suitability and efficiency.

Benefits of supplementing an industrial waste anaerobic digester with energy crops for increased biogas production.

PubMed

Nges, Ivo Achu; Escobar, Federico; Fu, Xinmei; Björnsson, Lovisa

2012-01-01

Currently, there is increasing competition for waste as feedstock for the growing number of biogas plants. This has led to fluctuation in feedstock supply and biogas plants being operated below maximum capacity. The feasibility of supplementing a protein/lipid-rich industrial waste (pig manure, slaughterhouse waste, food processing and poultry waste) mesophilic anaerobic digester with carbohydrate-rich energy crops (hemp, maize and triticale) was therefore studied in laboratory scale batch and continuous stirred tank reactors (CSTR) with a view to scale-up to a commercial biogas process. Co-digesting industrial waste and crops led to significant improvement in methane yield per ton of feedstock and carbon-to-nitrogen ratio as compared to digestion of the industrial waste alone. Biogas production from crops in combination with industrial waste also avoids the need for micronutrients normally required in crop digestion. The batch co-digestion methane yields were used to predict co-digestion methane yield in full scale operation. This was done based on the ratio of methane yields observed for laboratory batch and CSTR experiments compared to full scale CSTR digestion of industrial waste. The economy of crop-based biogas production is limited under Swedish conditions; therefore, adding crops to existing industrial waste digestion could be a viable alternative to ensure a constant/reliable supply of feedstock to the anaerobic digester. Copyright © 2011 Elsevier Ltd. All rights reserved.

The Physical State of Emulsified Edible Oil Modulates Its in Vitro Digestion.

PubMed

Guo, Qing; Bellissimo, Nick; Rousseau, Dérick

2017-10-18

Emulsified lipid digestion was tailored by manipulating the physical state of dispersed oil droplets in whey protein stabilized oil-in-water (O/W) emulsions, where the oil phase consisted of one of five ratios of soybean oil (SO) and fully hydrogenated soybean oil (FHSO). The evolution in particle size distribution, structural changes during oral, gastric, and intestinal digestion, and free fatty acid release during intestinal digestion were all investigated. Irrespective of the physical state and structure of the dispersed oil/fat, all emulsions were stable against droplet size increases during oral digestion. During gastric digestion, the 50:50 SO:FHSO emulsion was more stable against physical breakdown than any other emulsion. All emulsions underwent flocculation and coalescence or partial coalescence upon intestinal digestion, with the SO emulsion being hydrolyzed the most rapidly. The melting point of all emulsions containing FHSO was above 37 °C, with the presence of solid fat within the dispersed oil droplets greatly limiting lipolysis. Fat crystal polymorph and nanoplatelet size did not play an important role in the rate and extent of lipid digestion. Free fatty acid release modeled by the Weibull distribution function showed that the rate of lipid digestion (κ) decreased with increasing solid fat content, and followed an exponential relationship (R 2 = 0.95). Overall, lipid digestion was heavily altered by the physical state of the dispersed oil phase within O/W emulsions.

PCR-DGGE Analysis on Microbial Community Structure of Rural Household Biogas Digesters in Qinghai Plateau.

PubMed

Han, Rui; Yuan, Yongze; Cao, Qianwen; Li, Quanhui; Chen, Laisheng; Zhu, Derui; Liu, Deli

2018-05-01

To investigate contribution of environmental factor(s) to microbial community structure(s) involved in rural household biogas fermentation at Qinghai Plateau, we collected slurry samples from 15 digesters, with low-temperature working conditions (11.1-15.7 °C) and evenly distributed at three counties (Datong, Huangyuan, and Ledu) with cold plateau climate, to perform polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) and further sequencing. The bacterial communities in the total 15 digesters were classified into 38 genera with Mangroviflexus (12.1%) as the first dominant, and the archaeal communities into ten genera with Methanogenium (38.5%) as the most dominant. For each county, the digesters with higher biogas production, designated as HP digesters, exclusively had 1.6-3.1 °C higher fermentation temperature and the unique bacterial structure composition related, i.e., unclassified Clostridiales for all the HP digesters and unclassified Marinilabiliaceae and Proteiniclasticum for Ledu HP digesters. Regarding archaeal structure composition, Methanogenium exhibited significantly higher abundances at all the HP digesters and Thermogymnomonas was the unique species only identified at Ledu HP digesters with higher-temperature conditions. Redundancy analysis also confirmed the most important contribution of temperature to the microbial community structures investigated. This report emphasized the correlation between temperature and specific microbial community structure(s) that would benefit biogas production of rural household digesters at Qinghai Plateau.

Thermophilic anaerobic co-digestion of garbage, screened swine and dairy cattle manure.

PubMed

Liu, Kai; Tang, Yue-Qin; Matsui, Toru; Morimura, Shigeru; Wu, Xiao-Lei; Kida, Kenji

2009-01-01

Methane fermentation characteristics of garbage, swine manure (SM), dairy cattle manure (DCM) and mixtures of these wastes were studied. SM and DCM showed much lower volatile total solid (VTS) digestion efficiencies and methane yield than those of garbage. VTS digestion efficiency of SM was significantly increased when it was co-digested with garbage (Garbage: SM=1:1). Co-digestion of garbage, SM and DCM with respect to the relative quantity of each waste discharged in the Kikuchi (1: 16: 27) and Aso (1: 19: 12) areas indicated that co-digestion with garbage would improve the digestion characteristic of SM and DCM as far as the ratio of DCM in the wastes was maintained below a certain level. When the mixed waste (Garbage: SM: DCM=1:19:12) was treated using a thermophilic UAF reactor, methanogens responsible for the methane production were Methanoculleus and Methanosarcina species. Bacterial species in the phylum Firmicutes were dominant bacteria responsible for the digestion of these wastes. As the percentage of garbage in the mixed wastes used in this study was low (2-3%) and the digestion efficiency of DCM was obviously improved, the co-digestion of SM and DCM with limited garbage was a prospective method to treat the livestock waste effectively and was an attractive alternative technology for the construction of a sustainable environment and society in stock raising area.

Anaerobic digestion of selected Italian agricultural and industrial residues (grape seeds and leather dust): combined methane production and digestate characterization.

PubMed

Caramiello, C; Lancellotti, I; Righi, F; Tatàno, F; Taurino, R; Barbieri, L

2013-01-01

A combined experimental evaluation of methane production (obtained by anaerobic digestion) and detailed digestate characterization (with physical-chemical, thermo-gravimetric and mineralogical approaches) was conducted on two organic substrates, which are specific to Italy (at regional and national levels). One of the substrates was grape seeds, which have an agricultural origin, whereas the other substrate was vegetable-tanned leather dust, which has an industrial origin. Under the assumed experimental conditions of the performed lab-scale test series, the grape seed substrate exhibited a resulting net methane production of 175.0 NmL g volatile solids (VS)(-1); hence, it can be considered as a potential energy source via anaerobic digestion. Conversely, the net methane production obtained from the anaerobic digestion of the vegetable-tanned leather dust substrate was limited to 16.1 NmL gVS(-1). A detailed characterization of the obtained digestates showed that there were both nitrogen-containing compounds and complex organic compounds present in the digestate that was obtained from the mixture of leather dust and inoculum. As a general perspective of this experimental study, the application of diversified characterization analyzes could facilitate (1) a better understanding of the main properties of the obtained digestates to evaluate their potential valorization, and (2) a combination of the digestate characteristics with the corresponding methane productions to comprehensively evaluate the bioconversion process.

Archaeal and Bacterial Community Structure in an Anaerobic Digestion Reactor (Lagoon Type) Used for Biogas Production at a Pig Farm.

PubMed

Pampillón-González, Liliana; Ortiz-Cornejo, Nadia L; Luna-Guido, Marco; Dendooven, Luc; Navarro-Noya, Yendi E

2017-01-01

Biogas production from animal waste is an economically viable way to reduce environmental pollution and produce valuable products, i.e., methane and a nutrient-rich organic waste product. An anaerobic digestion reactor for biogas production from pig waste was sampled at the entrance, middle (digestion chamber), and exit of a digester, while the bacterial and archaeal community structure was studied by 16S rRNA gene metagenomics. The number of bacterial operational taxonomic units (OTU)-97% was 3-7 times larger than that of archaeal ones. Bacteria and Archaea found in feces of animals (e.g., Clostridiaceae, Lachnospiraceae, Ruminococcaceae, Methanosarcina, Methanolobus, Methanosaeta, and Methanospirillum) dominated the entrance of the digester. The digestion chamber was dominated by anaerobic sugar-fermenting OP9 bacteria and the syntrophic bacteria Candidatus Cloacamonas (Waste Water of Evry 1; WWE1). The methanogens dominant in the digestion chamber were the acetoclastic Methanosaeta and the hydrogenothrophic Methanoculleus and Methanospirillum. Similar bacterial and archaeal groups that dominated in the middle of the digestion chamber were found in the waste that left the digester. Predicted functions associated with degradation of xenobiotic compounds were significantly different between the sampling locations. The microbial community found in an anaerobic digestion reactor loaded with pig manure contained microorganisms with biochemical capacities related to the 4 phases of methane production. © 2017 S. Karger AG, Basel.

In vitro evaluation on neutral detergent fiber and cellulose digestion by post-ruminal microorganisms in goats.

PubMed

Jiao, Jinzhen; Wang, Pengpeng; He, Zhixiong; Tang, Shaoxun; Zhou, Chuanshe; Han, Xuefeng; Wang, Min; Wu, Duanqin; Kang, Jinhe; Tan, Zhiliang

2014-07-01

Post-ruminal digestion of fiber has received much less attention than its ruminal digestion. Using in vitro incubation techniques, the present study explored whether variations in fiber digestion occurred in different segments of the post-ruminal tract and whether fiber structure could influence its digestibility. A split plot design was conducted with gut segments (jejunum, ileum, cecum and colon) as main plot and substrates (neutral detergent fiber (NDF) and cellulose (CEL)) as subplot. With the same substrate, the final asymptotic gas volume (V(F)), gas production at t(i) (V(t(i)), digestibility, microbial crude protein (MCP), total bacteria number (TBN), total short-chain fatty acids (TSCFA) and xylanase in incocula from the cecum and colon exceeded (P < 0.01) those in incocula from the jejunum and ileum, while the NH3-N in the former was less (P < 0.01). For the same gut segment, the digestion of CEL was superior to NDF, as reflected in greater V(F), V(t(i)), maximum rate of gas production, digestibility, enzyme activities and SCFA but lower pH and NH3-N. The current results imply that the intestinal contents from the cecum and colon have greater potential to digest fiber than those from the jejunum and ileum, and CEL is more easily digested in the post-ruminal tract than NDF. © 2013 Society of Chemical Industry.

Digested wheat gluten inhibits binding between leptin and its receptor.

PubMed

Jönsson, Tommy; Memon, Ashfaque A; Sundquist, Kristina; Sundquist, Jan; Olsson, Stefan; Nalla, Amarnadh; Bauer, Mikael; Linse, Sara

2015-01-20

Leptin resistance is considered a primary risk factor for obesity. It has been hypothesized that dietary cereal grain protein could cause leptin resistance by preventing leptin from binding to its receptor. Non-degraded dietary wheat protein has been found in human serum at a mean level of 41 ng/mL. Here, we report our findings from testing whether enzymatically digested gluten from wheat prevents leptin from binding to the leptin receptor in vitro. Gluten from wheat was digested with pepsin and trypsin under physiological conditions. Pepsin and trypsin activity was removed from the gluten digest with a 10 kDa spin-filter or by heat treatment at 100°C for 30 min. Binding to the leptin receptor of leptin mixed with gluten digest at a series of concentrations was measured using surface plasmon resonance technology. Binding of the gluten digest to the leptin receptor was not detected. Spin-filtered gluten digest inhibited binding of leptin to the leptin receptor, with 50% inhibition at a gluten digest concentration of ~10 ng/mL. Heat-treated gluten digest did not inhibit leptin binding. Digested wheat gluten inhibits binding of leptin to the leptin receptor, with half-maximal inhibition at 10 ng/mL. The inhibition is significant at clinically relevant concentrations and could therefore serve as a novel pathway to investigate to understand the molecular basis of leptin resistance, obesity and associated disorders.

Improved design of anaerobic digesters for household biogas production in indonesia: one cow, one digester, and one hour of cooking per day.

PubMed

Usack, Joseph G; Wiratni, Wiratni; Angenent, Largus T

2014-01-01

A government-sponsored initiative in Indonesia to design and implement low-cost anaerobic digestion systems resulted in 21 full-scale systems with the aim to satisfy the cooking fuel demands of rural households owning at least one cow. The full-scale design consisted of a 0.3 m diameter PVC pipe, which was operated as a conventional plug-flow system. The system generated enough methane to power a cooking stove for ∼ 1 h. However, eventual clogging from solids accumulation inside the bioreactor proved to be a major drawback. Here, we improved the digester configuration to remedy clogging while maintaining system performance. Controlled experiments were performed using four 9-L laboratory-scale digesters operated at a temperature of 27 ± 1 °C, a volatile solids loading rate of 2.0 g VS · L(-1) · day(-1), and a 21-day hydraulic retention time. Two of the digesters were replicates of the original design (control digesters), while the other two digesters included internal mixing or effluent recycle (experimental digesters). The performance of each digester was compared based on methane yields, VS removal efficiencies, and steady-state solids concentrations during an operating period of 311 days. Statistical analyses revealed that internal mixing and effluent recycling resulted in reduced solids accumulation compared to the controls without diminishing methane yields or solids removal efficiencies.

Effects of the application of digestates from wet and dry anaerobic fermentation to Japanese paddy and upland soils on short-term nitrification.

PubMed

Sawada, Kozue; Toyota, Koki

2015-01-01

Wet and dry anaerobic fermentation processes are operated for biogas production from organic matter, resulting in wet and dry digestates as by-products, respectively. The application of these digestates to soil as fertilizer has increased in recent years. Therefore, we herein compared the effects of applying wet digestates (pH 8.2, C/N ratio 4.5), dry digestates (pH 8.8, C/N ratio 23.4), and a chemical fertilizer to Japanese paddy and upland soils on short-term nitrification under laboratory aerobic conditions. Chloroform-labile C, an indicator of microbial biomass, was only minimally affected by these applications, indicating that a small amount of labile N was immobilized by microbes. All applications led to rapid increases in NO3 -N contents in both soils, and ammonia-oxidizing bacteria, but not archaea may play a critical role in net nitrification in the amended soils. The net nitrification rates for both soils were the highest after the application of dry digestates, followed by wet digestates and then the chemical fertilizer in order of decreasing soil pH. These results suggest that the immediate effects of applying digestates, especially dry digestates with the highest pH, on nitrate leaching need to be considered when digestates are used as alternative fertilizers.

In Vitro Effects of Cooking Methods on Digestibility of Lipids and Formation of Cholesterol Oxidation Products in Pork

PubMed Central

Moon, Sung Sil

2014-01-01

This study investigated the effects of cooking methods on the digestibility of lipids and formation of cholesterol oxidation products (COPs) in pork, during in vitro human digestion. Pork patties were cooked using four different methods (oven cooking, pan frying, boiling, and microwaving), to an internal temperature of approximately 85℃. The digestibility of pork patties were then evaluated, using the in vitro human digestion model that simulated the composition (pH, minerals, surfaceactive components, and enzymes) of digestive juices in the human mouth, stomach, and small intestine. The total lipid digestibility was higher after microwave cooking, whereas pan-frying resulted in lower in vitro digestibility, compared to the other cooking methods. The microwaving method followed by in vitro digestion also showed significantly higher content of free fatty acids and thiobarbituric acid reactive substances (TBARS), compared to the other cooking methods; whereas, the pan frying and boiling methods showed the lowest. Cholesterol content was not significantly different among the cooked samples before, and after in vitro human digestion. The formation of COPs was significantly higher in the microwave-treated pork samples, compared to those cooked by the other methods, which was consistent with the trend for lipid peroxidation (TBARS). We propose that from the point of view of COPs formation and lipid oxidation, the pan-frying or boiling methods would be useful. PMID:26761168

210Polonium bioaccessibility assessment in algae for human consumption: An in vitro gastrointestinal digestion method.

PubMed

Desideri, Donatella; Meli, Maria Assunta; Roselli, Carla; Feduzi, Laura; Ugolini, Lucia

2017-01-01

The occurrence and mobility of natural radioactive element as 210 Polonium ( 210 Po) in 13 commercial algae consumed in Italy by humans were determined because the effects on human health need to take into account the bioavailability of these elements. The simulation of gastrointestinal (GIT) digestion was divided into three stages and was accomplished using three different artificial solutions: saliva, gastric, and synthetic bile-pancreas solution. The same sample was treated in two different ways: a) only gastric digestion and b) complete GIT digestion (gastric digestion followed by bile-pancreas solution). The difference between Po gastric mobility with respect to that found for GIT digestion was not significant; in fact, Po mobility exhibited a mean value 17.2 ± 15.1% and 19.5 ± 11.5% for gastric and GIT digestion, respectively.

Thermophilic biogasification of biomass

DOE Office of Scientific and Technical Information (OSTI.GOV)

Ghosh, S.; Klass, D.L.; Edwards, V.H.

1980-01-01

Secondary sewage effluent- and fresh-water-grown water hyacinths (Eichhornia crassipes), Coastal Bermuda grass (Cynodon dactylon), and a hyacinth-grass-municipal solid waste-sludge (biomass-waste) blend were used as test feeds to develop a fast thermophilic biomass- digestion process. For the pure biomass feeds thermophilic digestion has no apparent advantage over mesophilic digestion, but the reverse is true for the biomass-waste blend. Alkaline pretreatment of the feed improved thermophilic digester performance substantially. For a given plant feed load, the reactor volume, culture-heating requirements, and CH4 production rate for thermophilic digestion of the pretreated biomass-waste feed were 18,46, and 135% of those for conventional mesophilic digestion.more » For a biomass-waste feed the respective volatile solids reduction and energy recovery efficiencies were 46 and 49% for thermophilic and 36 and 43% for mesophilic digestions.« less

Modifications in trypsin digestion protocol for increasing the efficiency and coverage.

PubMed

Syal, Kirtimaan; Tadala, Raghu

2015-01-01

Standard trypsin digestion protocol of proteins followed by MALDI-MS analysis has been realized as an important tool for the identification and characterization of proteins. In this article, we proposed the elimination of the step of 'staining/de-staining of gel pieces' in in-gel digestion protocol in order to improve the efficiency of trypsin digestion. Coomassie dye is known to interfere with digestion of proteins by trypsin and the procedure of staining-de-staining could result in loss of photoaffinity probe, post translational modifications and catalytic activities of enzymes. Further, we studied parameters like hydrophobicity and isoelectric point, and attempted to quantitatively relate it to the efficiency of trypsin digestion. We suggest that properties of proteins should be considered and trypsin digestion protocol should be appropriately modified as per sequence and other information.

Anaerobic sludge digestion in the presence of lactobacillus additive

DOE Office of Scientific and Technical Information (OSTI.GOV)

Ghosh, S.; Klass, D.L.

1980-01-01

A laboratory evaluation of a lactobacillus fermentation product was performed to study its effects as an additive on the anaerobic digestion of sewage sludge under conventional and overloaded high-rate conditions. The overloaded conditions were those experienced in commercial municipal digesters. It was concluded from this work that the use of the additive at low concentrations permits digester operation at least up to double the loading of untreated digesters and at higher methane yields and volatile solids reductions without affecting effluent quality. The additive also imparts iproved digester stability and rapid response to loading rate and detention time excursions and upsets.more » The beneficial effets of the additive observed in the laboratory remain to be established with other feeds such as biomass, and in large-scale commercial digestion tests that are now in progress.« less

Potential for chlorate interference in ion chromatographic determination of total nitrogen in natural waters following alkaline persulfate digestion.

PubMed

Halstead, J A; Edwards, J; Soracco, R J; Armstrong, R W

1999-10-01

Determination of total nitrogen in aqueous samples after thermal potassium peroxydisulfate (persulfate) digestion is a commonly used alternative to the tedious Kjeldahl procedure. When ion chromatography is used to quantify the nitrate formed during digestion, there is a potential for interference from a chlorate peak if the digested sample initially contained chloride in concentrations close to or greater than the concentration of nitrogen. It was determined that this interference can be avoided either by using chromatographic conditions which cleanly resolve the nitrate and chlorate peaks (e.g., the Dionex AG9-HG column) or by using digestion reagent concentrations chosen to maintain a high pH throughout the digestion. The second alternative is not a viable option for investigators using a single digestion for both total nitrogen (TN) and total phosphorus (TP) analysis.

Application of Methanobrevibacter acididurans in anaerobic digestion.

PubMed

Savant, D V; Ranade, D R

2004-01-01

To operate anaerobic digesters successfully under acidic conditions, hydrogen utilizing methanogens which can grow efficiently at low pH and tolerate high volatile fatty acids (VFA) are desirable. An acid tolerant hydrogenotrophic methanogen viz. Methanobrevibacter acididurans isolated from slurry of an anaerobic digester running on alcohol distillery wastewater has been described earlier by this lab. This organism could grow optimally at pH 6.0. In the experiments reported herein, M. acididurans showed better methanogenesis under acidic conditions with high VFA, particularly acetate, than Methanobacterium bryantii, a common hydrogenotrophic inhabitant of anaerobic digesters. Addition of M. acididurans culture to digesting slurry of acidogenic as well as methanogenic digesters running on distillery wastewater showed increase in methane production and decrease in accumulation of volatile fatty acids. The results proved the feasibility of application of M. acididurans in anaerobic digesters.

Comparison of amino acid digestibility of feedstuffs determined with the precision-fed cecectomized rooster assay and the standardized ileal amino acid digestibility assay.

PubMed

Kim, E J; Utterback, P L; Applegate, T J; Parsons, C M

2011-11-01

The objective of this study was to evaluate and compare amino acid digestibility of several feedstuffs using 2 commonly accepted methods: the precision-fed cecectomized rooster assay (PFR) and the standardized ileal amino acid assay (SIAAD). Six corn, 6 corn distillers dried grains with or without solubles (DDGS/DDG), one wet distillers grains, one condensed solubles, 2 meat and bone meal (MBM) and a poultry byproduct meal were evaluated. Due to insufficient amounts, the wet distillers grains and condensed solubles were only evaluated in roosters. Standardized amino acid digestibility varied among the feed ingredients and among samples of the same ingredient for both methods. For corn, there were generally no differences in amino acid digestibility between the 2 methods. When differences did occur, there was no consistent pattern among the individual amino acids and methods. Standardized amino acid digestibility was not different between the 2 methods for 4 of the DDG samples; however, the PFR yielded higher digestibility values for a high protein DDG and a conventionally processed DDGS. The PFR yielded higher amino acid digestibility values than the SIAAD for several amino acids in 1 MBM and the poultry byproduct meal, but it yielded lower digestibility values for the other MBM. Overall, there were no consistent differences between methods for amino acid digestibility values. In conclusion, the PFR and SIAAD methods are acceptable for determining amino acid digestibility. However, these procedures do not always yield similar results for all feedstuffs evaluated. Thus, further studies are needed to understand the underlying causes in this variability.

Experimental co-digestion of corn stalk and vermicompost to improve biogas production

DOE Office of Scientific and Technical Information (OSTI.GOV)

Chen Guangyin; Zheng Zheng, E-mail: zzhenghj@fudan.edu.c; Department of Environmental Science and Engineering, Fudan University, Shanghai 200433

2010-10-15

Anaerobic co-digestion of corn stalk and vermicompost (VC) as well as mono-digestion of corn stalk were investigated. Batch mono-digestion experiments were performed at 35 {+-} 1 {sup o}C and initial total solid loading (TSL) ranged from 1.2% to 6.0%. Batch co-digestion experiments were performed at 35 {+-} 1 {sup o}C and initial TSL of 6% with VC proportions ranged from 20% to 80% of total solid (TS). For mono-digestion of corn stalk, a maximum methane yield of 217.60 {+-} 13.87 mL/g TS{sub added} was obtained at initial TSL of 4.8%, and acidification was found at initial TSL of 6.0% withmore » the lowest pH value of 5.10 on day 4. Co-digestion improved the methane yields by 4.42-58.61% via enhancing volatile fatty acids (VFAs) concentration and pH value compared with mono-digestion of corn stalk. The maximum biogas yield of 410.30 {+-} 11.01 mL/g TS{sub added} and methane yield of 259.35 {+-} 13.85 mL/g TS{sub added} were obtained for 40% VC addition. Structure analysis by X-ray diffractometry (XRD) showed that the lowest crystallinity of 35.04 of digested corn stalk was obtained from co-digestion with 40% VC, which decreased 29.4% compared to 49.6 obtained from un-treated corn stalk. It is concluded that co-digestion with VC is beneficial for improving biodigestibility and methane yield from corn stalk.« less

Design of lipid-based formulations for oral administration of poorly water-soluble drug fenofibrate: effects of digestion.

PubMed

Mohsin, Kazi

2012-06-01

Lipid-based drug carriers are likely to have influence on bioavailability through enhanced solubilization of the drug in the gastrointestinal tract. The study was designed to investigate the lipid formulation digestibility in the simulated gastro intestinal media. Fenofibrate was formulated in representative Type II, IIIA, IIIB and IV self-emulsifying/microemulsifying lipid delivery systems (SEDDS and SMEDDS designed for oral administration) using various medium-chain glyceride components, non-ionic surfactants and cosolvents as excipients. Soybean oil was used only as an example of long-chain triglycerides to compare the effects of formulation with their counterparts. The formulations were subjected to in vitro digestion specifically to predict the fate of the drug in the gastro intestinal tract after exposure of the formulation to pancreatic enzymes and bile. In vitro digestion experiments were carried out using a pH-stat maintained at pH 7.5 for 30 min using intestinal fluids simulating the fed and fasted states. The digestion rate was faster and almost completed in Type II and IIIA systems. Most of the surfactants used in the studies are digestible. However, the high concentration of surfactant and/or cosolvent used in Type IIIB or IV systems lowered the rate of digestion. The digestion of medium-chain triglycerides was faster than long-chain triglycerides, but kept comparatively less drug in the post digestion products. Medium-chain mixed glycerides are good solvents for fenofibrate as rapidly digested but to improve fenofibrate concentration in post digestion products the use of long-chain mixed glycerides are suggested for further investigations.

The on-bead digestion of protein corona on nanoparticles by trypsin immobilized on the magnetic nanoparticle.

PubMed

Hu, Zhengyan; Zhao, Liang; Zhang, Hongyan; Zhang, Yi; Wu, Ren'an; Zou, Hanfa

2014-03-21

Proteins interacting with nanoparticles would form the protein coronas on the surface of nanoparticles in biological systems, which would critically impact the biological identities of nanoparticles and/or result in the physiological and pathological consequences. The enzymatic digestion of protein corona was the primary step to achieve the identification of protein components of the protein corona for the bottom-up proteomic approaches. In this study, the investigation on the tryptic digestion of protein corona by the immobilized trypsin on a magnetic nanoparticle was carried out for the first time. As a comparison with the usual overnight long-time digestion and the severe self-digestion of free trypsin, the on-bead digestion of protein corona by the immobilized trypsin could be accomplished within 1h, along with the significantly reduced self-digestion of trypsin and the improved reproducibility on the identification of proteins by the mass spectrometry-based proteomic approach. It showed that the number of identified bovine serum (BS) proteins on the commercial Fe3O4 nanoparticles was increased by 13% for the immobilized trypsin with 1h digestion as compared to that of using free trypsin with even overnight digestion. In addition, the on-bead digestion of using the immobilized trypsin was further applied on the identification of human plasma protein corona on the commercial Fe3O4 nanoparticles, which leads the efficient digestion of the human plasma proteins and the identification of 149 human plasma proteins corresponding to putative critical pathways and biological processes. Copyright © 2014 Elsevier B.V. All rights reserved.

Bioaugmentation of overloaded anaerobic digesters restores function and archaeal community.

PubMed

Tale, V P; Maki, J S; Zitomer, D H

2015-03-01

Adding beneficial microorganisms to anaerobic digesters for improved performance (i.e. bioaugmentation) has been shown to decrease recovery time after organic overload or toxicity upset. Compared to strictly anaerobic cultures, adding aerotolerant methanogenic cultures may be more practical since they exhibit higher methanogenic activity and can be easily dried and stored in ambient air for future shipping and use. In this study, anaerobic digesters were bioaugmented with both anaerobic and aerated, methanogenic propionate enrichment cultures after a transient organic overload. Digesters bioaugmented with anaerobic and moderately aerated cultures recovered 25 and 100 days before non-bioaugmented digesters, respectively. Increased methane production due to bioaugmentation continued a long time, with 50-120% increases 6 to 12 SRTs (60-120 days) after overload. In contrast to the anaerobic enrichment, the aerated enrichments were more effective as bioaugmentation cultures, resulting in faster recovery of upset digester methane and COD removal rates. Sixty days after overload, the bioaugmented digester archaeal community was not shifted, but was restored to one similar to the pre-overload community. In contrast, non-bioaugmented digester archaeal communities before and after overload were significantly different. Organisms most similar to Methanospirillum hungatei had higher relative abundance in well-operating, undisturbed and bioaugmented digesters, whereas organisms similar to Methanolinea tarda were more abundant in upset, non-bioaugmented digesters. Bioaugmentation is a beneficial approach to increase digester recovery rate after transient organic overload events. Moderately aerated, methanogenic propionate enrichment cultures were more beneficial augments than a strictly anaerobic enrichment. Copyright © 2014 Elsevier Ltd. All rights reserved.

A Research-Oriented Approach to Digestive Physiology To Replace Traditional Enzymatic Laboratories.

ERIC Educational Resources Information Center

Grabowski, Gregory M.; Holt, Jelena

2002-01-01

Describes a physiology laboratory designed to localize digestive enzymes within the digestive tract of cockroaches and develop a general conclusion about the similarities to mammalian digestion. This approach not only demonstrates the practicality of lecture material, but also provides a springboard for independent research opportunities.…

78 FR 56906 - National Institute of Diabetes and Digestive and Kidney Diseases; Notice of Meetings

Federal Register 2010, 2011, 2012, 2013, 2014

2013-09-16

... Diabetes and Digestive and Kidney Diseases; Notice of Meetings Pursuant to section 10(d) of the Federal... Digestive and Kidney Diseases Initial Review Group Diabetes, Endocrinology and Metabolic Diseases B..., [email protected] . Name of Committee: National Institute of Diabetes and Digestive and Kidney...

76 FR 20692 - National Institute of Diabetes and Digestive and Kidney Diseases; Notice of Meeting

Federal Register 2010, 2011, 2012, 2013, 2014

2011-04-13

... Diabetes and Digestive and Kidney Diseases; Notice of Meeting Pursuant to section 10(d) of the Federal... Digestive and Kidney Diseases Initial Review Group; Diabetes, Endocrinology and Metabolic Diseases B... and Metabolic Research; 93.848, Digestive Diseases and Nutrition Research; 93.849, Kidney Diseases...

77 FR 53208 - National Institute of Diabetes and Digestive and Kidney Diseases; Notice of Closed Meeting

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2012-08-31

... Diabetes and Digestive and Kidney Diseases; Notice of Closed Meeting Pursuant to section 10(d) of the... of Diabetes and Digestive And Kidney Diseases, including consideration of personnel qualifications..., Ph.D., Scientific Director, National Institute of Diabetes and Digestive and Kidney Diseases...

76 FR 30733 - National Institute of Diabetes and Digestive and Kidney Diseases; Notice of Meeting

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2011-05-26

... Diabetes and Digestive and Kidney Diseases; Notice of Meeting Pursuant to section 10(d) of the Federal... Digestive and Kidney Diseases Initial Review Group, Digestive Diseases and Nutrition C Subcommittee. Date... Diseases and Nutrition Research; 93.849, Kidney Diseases, Urology and Hematology Research, National...

77 FR 33750 - National Institute of Diabetes and Digestive and Kidney Diseases; Notice of Closed Meeting

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2012-06-07

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78 FR 58325 - National Institute of Diabetes and Digestive and Kidney Diseases; Notice of Closed Meetings

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2013-09-23

... Diabetes and Digestive and Kidney Diseases; Notice of Closed Meetings Pursuant to section 10(d) of the... Diabetes and Digestive and Kidney Diseases Special Emphasis Panel, Bariatric Surgery-- Related Ancillary... of Diabetes and Digestive and Kidney Diseases Special Emphasis Panel, Regulatory Mechanisms in...

Effect of temperature on methane production from field-scale anaerobic digesters treating dairy manure

USDA-ARS?s Scientific Manuscript database

Temperature is a critical factor affecting anaerobic digestion because it influences both system heating requirements and methane production. Temperatures of 35-37°C are typically suggested for manure digestion, yet in temperate climate digesters, require a considerable amount of additional heat en...

40 CFR 428.90 - Applicability; description of the pan, dry digestion, and mechanical reclaimed rubber subcategory.

Code of Federal Regulations, 2013 CFR

2013-07-01

..., dry digestion, and mechanical reclaimed rubber subcategory. 428.90 Section 428.90 Protection of... MANUFACTURING POINT SOURCE CATEGORY Pan, Dry Digestion, and Mechanical Reclaimed Rubber Subcategory § 428.90 Applicability; description of the pan, dry digestion, and mechanical reclaimed rubber subcategory. The...

40 CFR 428.90 - Applicability; description of the pan, dry digestion, and mechanical reclaimed rubber subcategory.

Code of Federal Regulations, 2012 CFR

2012-07-01

..., dry digestion, and mechanical reclaimed rubber subcategory. 428.90 Section 428.90 Protection of... MANUFACTURING POINT SOURCE CATEGORY Pan, Dry Digestion, and Mechanical Reclaimed Rubber Subcategory § 428.90 Applicability; description of the pan, dry digestion, and mechanical reclaimed rubber subcategory. The...

40 CFR 428.90 - Applicability; description of the pan, dry digestion, and mechanical reclaimed rubber subcategory.

Code of Federal Regulations, 2011 CFR

2011-07-01

..., dry digestion, and mechanical reclaimed rubber subcategory. 428.90 Section 428.90 Protection of... POINT SOURCE CATEGORY Pan, Dry Digestion, and Mechanical Reclaimed Rubber Subcategory § 428.90 Applicability; description of the pan, dry digestion, and mechanical reclaimed rubber subcategory. The...

40 CFR 428.90 - Applicability; description of the pan, dry digestion, and mechanical reclaimed rubber subcategory.

Code of Federal Regulations, 2014 CFR

2014-07-01

..., dry digestion, and mechanical reclaimed rubber subcategory. 428.90 Section 428.90 Protection of... MANUFACTURING POINT SOURCE CATEGORY Pan, Dry Digestion, and Mechanical Reclaimed Rubber Subcategory § 428.90 Applicability; description of the pan, dry digestion, and mechanical reclaimed rubber subcategory. The...

40 CFR 428.90 - Applicability; description of the pan, dry digestion, and mechanical reclaimed rubber subcategory.

Code of Federal Regulations, 2010 CFR

2010-07-01

..., dry digestion, and mechanical reclaimed rubber subcategory. 428.90 Section 428.90 Protection of... POINT SOURCE CATEGORY Pan, Dry Digestion, and Mechanical Reclaimed Rubber Subcategory § 428.90 Applicability; description of the pan, dry digestion, and mechanical reclaimed rubber subcategory. The...

76 FR 10042 - National Institute of Diabetes and Digestive and Kidney Diseases; Notice of Closed Meetings

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2011-02-23

... Digestive and Kidney Diseases Special Emphasis Panel, PAR09-247 Ancillary Clinical Studies of Interest to... Institute of Diabetes and Digestive and Kidney Diseases Special Emphasis Panel, Nutrition and Metabolism.... 93.847, Diabetes, Endocrinology and Metabolic Research; 93.848, Digestive Diseases and Nutrition...

Methane and hydrogen sulfide production during co-digestion of forage radish and dairy manure

USDA-ARS?s Scientific Manuscript database

Forage radish cover crops were investigated as a co-substrate to increase biogas production from dairy manure-based anaerobic digestion. Lab-scale batch digesters (300 mL) were operated under mesophilic conditions during two experiments. In the first experiment, the optimal co-digestion ratio for ...

Developing a Decision Support Tool for Waste to Energy Calculations Using Energy Return on Investment

DTIC Science & Technology

2016-12-01

Incinerator with Cogeneration. Source: Taylor (2016). 2. Anaerobic Digestion Anaerobic digestion uses a fermentation process to produce methane from...ANAEROBIC DIGESTION Anaerobic digestion uses a fermentation process to produce methane from organic waste inputs, resulting in a biogas that is then

Supervision: Exploring the Effective Components. ERIC/CASS Counseling Digest Series.

ERIC Educational Resources Information Center

Borders, L. DiAnne, Ed.

This document contains a collection of ERIC Digests on supervision, a topic of critical professional importance for counselors. Following an introductory article by the guest editor, L. DiAnne Borders, "Supervision: Exploring the Effective Components," 19 digests address a different facet of supervision. The 19 digests are: (1)…

Anaerobic digestion of organic solid poultry slaughterhouse waste--a review.

PubMed

Salminen, E; Rintala, J

2002-05-01

This work reviews the potential of anaerobic digestion for material recovery and energy production from poultry slaughtering by-products and wastes. First, we describe and quantify organic solid by-products and wastes produced in poultry farming and poultry slaughterhouses and discuss their recovery and disposal options. Then we review certain fundamental aspects of anaerobic digestion considered important for the digestion of solid slaughterhouse wastes. Finally, we present an overview of the future potential and current experience of the anaerobic digestion treatment of these materials.

Ascorbate and Apple Phenolics Affect Protein Oxidation in Emulsion-Type Sausages during Storage and in Vitro Digestion.

PubMed

Rysman, Tine; Van Hecke, Thomas; De Smet, Stefaan; Van Royen, Geert

2016-05-25

The effect of sodium ascorbate and apple phenolics on the oxidative stability of emulsion-type sausages during storage and digestion was investigated. Emulsion-type sausages containing 0.05% sodium ascorbate or 3% freeze-dried apple pomace were subjected to chilled illuminated storage and subsequent in vitro digestion. Lipid oxidation was assessed as TBARS, and protein oxidation was evaluated as thiol oxidation, total carbonyls, and γ-glutamic and α-amino adipic semialdehyde. Proteolysis was measured after digestion to evaluate protein digestibility. The results suggest the presence of protein-ascorbate and protein-phenol interactions, which may decrease protein digestibility and may interfere with spectrophotometric methods for measuring oxidation.

Anaerobic digestion of slaughterhouse waste with UF-membrane separation and recycling of permeate after free ammonia stripping.

PubMed

Siegrist, H; Hunziker, W; Hofer, H

2005-01-01

Anaerobic digestion can adapt to free ammonia to a certain extent. During the anaerobic digestion of slaughterhouse waste, however, an ammonia concentration of up to 15 g Nl(-1) can be reached in the sludge liquid and this will even inhibit adapted sludge. To lower this concentration, a fraction of the digester liquid must therefore be continuously separated from the digested sludge and the free ammonia stripped before the liquid is recycled to the digester. A mesophilic laboratory digester was successfully operated with an ammonium concentration of 4-5g l(-1) and a pH of 8.0-8.4. After free ammonia stripping, the excess liquid was treated in a laboratory SBR for nitrogen and phosphorus removal before being added to the receiving water. The effluent had no toxic effect on daphnia and algae.

Development of an advanced anaerobic digester design and a kinetic model for biogasification of water hyacinth/sludge blends

DOE Office of Scientific and Technical Information (OSTI.GOV)

Srivastava, V.; Fannin, K.F.; Biljetina, R.

1986-07-01

The Institute of Gas Technology (IGT) conducted a comprehensive laboratory-scale research program to develop and optimize the anaerobic digestion process for producing methane from water hyacinth and sludge blends. This study focused on digester design and operating techniques, which gave improved methane yields and production rates over those observed using conventional digesters. The final digester concept and the operating experience was utilized to design and operate a large-scale experimentla test unit (ETU) at Walt Disney World, Florida. This paper describes the novel digester design, operating techniques, and the results obtained in the laboratory. The paper also discusses a kinetic modelmore » which predicts methane yield, methane production rate, and digester effluent solids as a function of retention time. This model was successfully utilized to predict the performance of the ETU. 15 refs., 6 figs., 6 tabs.« less

Anaerobic co-digestion of municipal organic wastes and pre-treatment to enhance biogas production from waste.

PubMed

Li, Chenxi; Champagne, Pascale; Anderson, Bruce C

2014-01-01

Co-digestion and pre-treatment have been recognized as effective, low-cost and commercially viable approaches to reduce anaerobic digestion process limitations and improve biogas yields. In our previous batch-scale study, fat, oil, and grease (FOG) was investigated as a suitable potential co-substrate, and thermo-chemical pre-treatment (TCPT) at pH = 10 and 55 °C improved CH4 production from FOG co-digestions. In this project, co-digestions with FOG were studied in bench-scale two-stage thermophilic semi-continuous flow co-digesters with suitable TCPT (pH = 10, 55 °C). Overall, a 25.14 ± 2.14 L/d (70.2 ± 1.4% CH4) biogas production was obtained, which was higher than in the two-stage system without pre-treatment. The results could provide valuable fundamental information to support full-scale investigations of anaerobic co-digestion of municipal organic wastes.

Combined anaerobic and aerobic digestion for increased solids reduction and nitrogen removal.

PubMed

Novak, John T; Banjade, Sarita; Murthy, Sudhir N

2011-01-01

A unique sludge digestion system consisting of anaerobic digestion followed by aerobic digestion and then a recycle step where thickened sludge from the aerobic digester was recirculated back to the anaerobic unit was studied to determine the impact on volatile solids (VS) reduction and nitrogen removal. It was found that the combined anaerobic/aerobic/anaerobic (ANA/AER/ANA) system provided 70% VS reduction compared to 50% for conventional mesophilic anaerobic digestion with a 20 day SRT and 62% for combined anaerobic/aerobic (ANA/AER) digestion with a 15 day anaerobic and a 5 day aerobic SRT. Total Kjeldahl nitrogen (TKN) removal for the ANA/AER/ANA system was 70% for sludge wasted from the aerobic unit and 43.7% when wasted from the anaerobic unit. TKN removal was 64.5% for the ANA/AER system. Copyright © 2010 Elsevier Ltd. All rights reserved.

Anaerobic sludge digestion with a biocatalytic additive. [Lactobacillus acidophilus

DOE Office of Scientific and Technical Information (OSTI.GOV)

Ghosh, S.; Henry, M.P.; Fedde, P.A.

1982-01-01

Aimed at improving the process operating characteristics of anaerobic digestion for sludge stabilization and SNG production, this study evaluates the effects of a lactobacillus additive under normal, variable, and overload conditions. This whey fermentation product of an acid-tolerant strain of L. acidophilus fortified with CoCO/sub 3/, (NH/sub 4/)/sub 2/HPO/sub 4/, ferrous lactate, and lactic acid provides growth factors, metabolic intermediates, and enzymes needed for substrate degradation and cellular synthesis. Data indicate that the biochemical additive increases methane yield, gas production rate, and volatile solids reduction; decreases volatile acids accumulation; enhances the digester buffer capacity; and improves the fertilizer value andmore » dewatering characteristics of the digested residue. Digester capacities could be potentially doubled when the feed is so treated. Results of field tests with six full-scale digesters confirm observations made with bench-scale digesters.« less

77 FR 50518 - National Institute of Diabetes and Digestive and Kidney Diseases; Notice of Meetings

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2012-08-21

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78 FR 10623 - National Institute of Diabetes and Digestive and Kidney Diseases; Notice of Closed Meeting

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2013-02-14

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78 FR 26641 - National Institute of Diabetes and Digestive and Kidney Diseases; Notice of Closed Meeting

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2013-05-07

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2013-11-22

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78 FR 48456 - National Institute of Diabetes and Digestive and Kidney Diseases; Notice of Closed Meetings

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2013-08-08

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2012-11-07

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76 FR 10042 - National Institute of Diabetes and Digestive and Kidney Diseases; Notice of Closed Meetings

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2011-02-23

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76 FR 63313 - National Institute of Diabetes and Digestive and Kidney Diseases; Notice of Closed Meeting

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2011-10-12

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76 FR 36931 - National Institute of Diabetes and Digestive and Kidney Diseases; Notice of Closed Meetings

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2011-06-23

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75 FR 61766 - National Institute of Diabetes and Digestive and Kidney Diseases; Notice of Closed Meetings

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2010-10-06

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75 FR 38818 - National Institute of Diabetes and Digestive and Kidney Diseases; Notice of Closed Meetings

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2010-07-06

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78 FR 5467 - National Institute of Diabetes and Digestive and Kidney Diseases; Notice of Closed Meeting

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2013-01-25

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77 FR 2076 - National Institute of Diabetes and Digestive and Kidney Diseases Notice of Closed Meetings

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2012-01-13

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2010-06-23

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75 FR 25278 - National Institute of Diabetes and Digestive and Kidney Diseases; Notice of Closed Meetings

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78 FR 55086 - National Institute of Diabetes and Digestive and Kidney Diseases; Notice of Meeting

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76 FR 16432 - National Institute of Diabetes and Digestive and Kidney Diseases; Notice of Meetings

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78 FR 72684 - National Institute of Diabetes and Digestive and Kidney Diseases; Notice of Closed Meeting

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2013-12-03

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77 FR 12855 - National Institute of Diabetes and Digestive and Kidney Diseases; Notice of Closed Meeting

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2012-03-02

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75 FR 3741 - National Institute of Diabetes and Digestive and Kidney Diseases; Notice of Meetings

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2010-01-22

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75 FR 4830 - National Institute of Diabetes and Digestive and Kidney Diseases;

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78 FR 76632 - National Institute of Diabetes and Digestive and Kidney Diseases; Notice of Meetings

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2013-12-18

... Diabetes and Digestive and Kidney Diseases; Notice of Meetings Pursuant to section 10(d) of the Federal... Diabetes and Digestive and Kidney Diseases Advisory Council. The meetings will be open to the public as... personal privacy. Name of Committee: National Diabetes and Digestive and Kidney Diseases Advisory Council...

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2011-05-26

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2010-11-15

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2013-09-09

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2011-12-15

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2013-02-07

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2010-10-22

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2011-05-03

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2010-03-10

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2012-02-17

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2010-03-26

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77 FR 33752 - National Institute of Diabetes and Digestive and Kidney Diseases; Notice of Closed Meetings

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2012-06-07

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2012-08-09

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2010-05-07

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78 FR 28859 - National Institute of Diabetes and Digestive and Kidney Diseases Notice of Closed Meetings

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2013-05-16

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77 FR 2075 - National Institute of Diabetes and Digestive and Kidney Diseases Notice of Meetings

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2012-01-13

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75 FR 56117 - National Institute of Diabetes and Digestive and Kidney Diseases; Notice of Meetings

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2010-09-15

... Diabetes and Digestive and Kidney Diseases; Notice of Meetings Pursuant to section 10(d) of the Federal... Digestive and Kidney Diseases Initial Review Group, Kidney, Urologic and Hematologic Diseases D Subcommittee.... [email protected] . Name of Committee: National Institute of Diabetes and Digestive and Kidney...

77 FR 6129 - National Institute of Diabetes and Digestive and Kidney Diseases Notice of Closed Meetings

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2012-02-07

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77 FR 28890 - National Institute of Diabetes and Digestive and Kidney Diseases Notice of Meetings

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2012-05-16

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2011-07-05

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77 FR 52750 - National Institute of Diabetes and Digestive and Kidney Diseases; Notice of Closed Meetings

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2012-08-30

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2011-06-07

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75 FR 49940 - National Institute of Diabetes and Digestive and Kidney Diseases; Notice of Meetings

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2010-08-16

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2013-05-16

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2010-03-04

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2010-03-01

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2013-10-22

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2011-07-29

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2011-03-17

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2011-03-17

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2011-03-01

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2013-04-15

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2012-02-22

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[Specific features of digestive function development in larvae of some salmonid fish].

PubMed

Ershova, T S; Volkova, I V; Zaĭtseva, V F

2004-01-01

We studied the activities of digestive enzymes responsible for the digestion of food carbohydrate and protein components in plant-eating fish at various stages of larval development. The activities of all digestive enzymes tend to rise during larval development. Species specific features of the alimentary canal functioning have been described.

75 FR 63495 - National Institute of Diabetes and Digestive and Kidney Diseases; Notice of Closed Meetings

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2010-10-15

... Diabetes and Digestive and Kidney Diseases Special Emphasis Panel, Kidney Diseases in Children Ancillary... Diabetes and Digestive and Kidney Diseases; Notice of Closed Meetings Pursuant to section 10(d) of the... Diabetes and Digestive and Kidney Diseases Special Emphasis Panel, DDK-C Conflicts. Date: November 16, 2010...

76 FR 33322 - National Institute of Diabetes and Digestive and Kidney Diseases; Notice of Closed Meetings

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2011-06-08

... Diabetes and Digestive and Kidney Diseases; Notice of Closed Meetings Pursuant to section 10(d) of the... Diabetes and Digestive and Kidney Diseases Special Emphasis Panel, NIDDK KUH-Fellowship Review. Date: June... . Name of Committee: National Institute of Diabetes and Digestive and Kidney Diseases Special Emphasis...

78 FR 13360 - National Institute of Diabetes and Digestive and Kidney Diseases; Notice of Closed Meeting

Federal Register 2010, 2011, 2012, 2013, 2014

2013-02-27

... Diabetes and Digestive and Kidney Diseases Special Emphasis Panel; Chronic Kidney Disease in Children. Date... Diabetes and Digestive and Kidney Diseases; Notice of Closed Meeting Pursuant to section 10(d) of the..., Diabetes, Endocrinology and Metabolic Research; 93.848, Digestive Diseases and Nutrition Research; 93.849...

78 FR 3903 - National Institute of Diabetes and Digestive and Kidney Diseases; Notice of Meetings

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2013-01-17

... Diabetes and Digestive and Kidney Diseases; Notice of Meetings Pursuant to section 10(d) of the Federal... Diabetes and Digestive and Kidney Diseases Advisory Council. The meetings will be open to the public as... privacy. Name of Committee: National Diabetes and Digestive and Kidney Diseases Advisory Council. Date...

78 FR 48455 - National Institute of Diabetes and Digestive and Kidney Diseases; Notice of Meetings

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2013-08-08

... Diabetes and Digestive and Kidney Diseases; Notice of Meetings Pursuant to section 10(d) of the Federal... Diabetes and Digestive and Kidney Diseases Advisory Council. The meetings will be open to the public as... privacy. Name of Committee: National Diabetes and Digestive and Kidney Diseases Advisory Council. Date...

78 FR 18358 - National Institute of Diabetes and Digestive and Kidney Diseases; Notice of Closed Meetings

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2013-03-26

... Diabetes and Digestive and Kidney Diseases Special Emphasis Panel; Review of U34 Clinical Trial Planning... of Diabetes and Digestive and Kidney Diseases Special Emphasis Panel; Review of U34 Clinical Trial... Metabolic Research; 93.848, Digestive Diseases and Nutrition Research; 93.849, Kidney Diseases, Urology and...

77 FR 76056 - National Institute of Diabetes and Digestive and Kidney Diseases; Notice of Closed Meetings

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2012-12-26

... Diabetes and Digestive and Kidney Diseases Special Emphasis Panel; Renal Supportive Care Studies. Date... Diabetes and Digestive and Kidney Diseases; Notice of Closed Meetings Pursuant to section 10(d) of the... Committee: National Institute of Diabetes and Digestive and Kidney Diseases Special Emphasis Panel; Diverse...

Recovery of ammonia in raw and co-digested swine manure using gas-permeable membrane technology

USDA-ARS?s Scientific Manuscript database

Anaerobic digestion of agro-industrial and livestock waste generates considerable digestate volumes that are important sources of nitrogen (N). However, on some occasions, the high concentrations of N present in the digestates may represent an obstacle to its use locally as fertilizer, since it can ...

Anaerobic co-digestion technology in solid wastes treatment for biomethane generation

NASA Astrophysics Data System (ADS)

Al Mamun, Muhammad Rashed; Torii, Shuichi

2017-05-01

Anaerobic co-digestion is considered to be an efficient way of disposing solid wastes which can not only reduce environmental burden, but also produce bioenergy. Co-digestion of solid wastes in the absence of bacteria inoculums with variable mixing ratios of three wastes has been experimentally tested for 35 days digestion time to determine the biogas potential. The temperature remained relatively constant at a mesophilic range of 29-36°C throughout the study. An average pH of 7.4 was recorded from all digesters. The average biogas yields obtained from the four digesters (D1, D2, D3 and D4) were 13.31, 15.67, 16.52 and 19.12 L/day, respectively. The cumulative result showed that from co-digestion of D4 43.67%, 22.02% and 15.71% more biogas was produced, respectively, than others. The maximum and average COD reduction was 57% and 31%, respectively, in co-digestion wastes. The biogas comprised average of 61% CH4, 33.5% CO2, 222 ppm H2S, and 4.7% H2O, respectively.

Exposure to polystyrene nanoplastic leads to inhibition of anaerobic digestion system.

PubMed

Fu, Shan-Fei; Ding, Jian-Nan; Zhang, Yun; Li, Yi-Fei; Zhu, Rong; Yuan, Xian-Zheng; Zou, Hua

2018-06-01

In this study, impacts of nanoplastic on the pure and mixed anaerobic digestion systems were investigated. Results showed the growth and metabolism of Acetobacteroides hydrogenigenes were partly inhibited by nanoplastic existed in the pure anaerobic digestion system. The anaerobic digestion of sewage sludge was also obviously inhibited by nanoplastic existed in the mixed anaerobic digestion system. Both the methane yield and methane production rate of the mixed anaerobic digestion system showed negative correlation with the nanoplastic concentration. Compared with anaerobic digestion system without nanoplastic, methane yield and maximum daily methane yield at the nanoplastic concentration of 0.2g/L decreased for 14.4% and 40.7%, respectively. In addition, the start-up of mixed anaerobic digestion system was prolonged by addition of nanoplastic. Microbial community structure analysis indicated the microbial community structures were also affected by nanoplastic existed in the system. At the nanoplastic concentration of 0.2g/L, the relative abundances of family Cloacamonaceae, Porphyromonadaceae, Anaerolinaceae and Gracilibacteraceae decreased partly. Conversely, the relative abundances of family Anaerolinaceae, Clostridiaceae, Geobacteraceae, Dethiosulfovibrionaceae and Desulfobulbaceae improved partly. Copyright © 2017 Elsevier B.V. All rights reserved.

Nanobiocatalysis for protein digestion in proteomic analysis

DOE Office of Scientific and Technical Information (OSTI.GOV)

Kim, Jungbae; Kim, Byoung Chan; Lopez-Ferrer, Daniel

2010-02-01

The process of protein digestion is a critical step for successful protein identification in the bottom-up proteomic analysis. To substitute the present practice of in-solution protein digestion, which is long, tedious, and difficult to automate, a lot of efforts have been dedicated for the development of a rapid, recyclable and automated digestion system. Recent advances of nanobiocatalytic approaches have improved the performance of protein digestion by using various nanomaterials such as nanoporous materials, magnetic nanoparticles, and polymer nanofibers. Especially, the unprecedented success of trypsin stabilization in the form of trypsin-coated nanofibers, showing no activity decrease under repeated uses for onemore » year and retaining good resistance to proteolysis, has demonstrated its great potential to be employed in the development of automated, high-throughput, and on-line digestion systems. This review discusses recent developments of nanobiocatalytic approaches for the improved performance of protein digestion in speed, detection sensitivity, recyclability, and trypsin stability. In addition, we also introduce the protein digestions under unconventional energy inputs for protein denaturation and the development of microfluidic enzyme reactors that can benefit from recent successes of these nanobiocatalytic approaches.« less

Resistance of Soil-Bound Prions to Rumen Digestion

PubMed Central

Saunders, Samuel E.; Bartelt-Hunt, Shannon L.; Bartz, Jason C.

2012-01-01

Before prion uptake and infection can occur in the lower gastrointestinal system, ingested prions are subjected to anaerobic digestion in the rumen of cervids and bovids. The susceptibility of soil-bound prions to rumen digestion has not been evaluated previously. In this study, prions from infectious brain homogenates as well as prions bound to a range of soils and soil minerals were subjected to in vitro rumen digestion, and changes in PrP levels were measured via western blot. Binding to clay appeared to protect noninfectious hamster PrPc from complete digestion, while both unbound and soil-bound infectious PrPSc proved highly resistant to rumen digestion. In addition, no change in intracerebral incubation period was observed following active rumen digestion of unbound hamster HY TME prions and HY TME prions bound to a silty clay loam soil. These results demonstrate that both unbound and soil-bound prions readily survive rumen digestion without a reduction in infectivity, further supporting the potential for soil-mediated transmission of chronic wasting disease (CWD) and scrapie in the environment. PMID:22937149

Cooking Chicken Breast Reduces Dialyzable Iron Resulting from Digestion of Muscle Proteins

PubMed Central

Gokhale, Aditya S.; Mahoney, Raymond R.

2014-01-01

The purpose of this research was to study the effect of cooking chicken breast on the production of dialyzable iron (an in vitro indicator of bioavailable iron) from added ferric iron. Chicken breast muscle was cooked by boiling, baking, sautéing, or deep-frying. Cooked samples were mixed with ferric iron and either extracted with acid or digested with pepsin and pancreatin. Total and ferrous dialyzable iron was measured after extraction or digestion and compared to raw chicken samples. For uncooked samples, dialyzable iron was significantly enhanced after both extraction and digestion. All cooking methods led to markedly reduced levels of dialyzable iron both by extraction and digestion. In most cooked, digested samples dialyzable iron was no greater than the iron-only (no sample) control. Cooked samples showed lower levels of histidine and sulfhydryls but protein digestibility was not reduced, except for the sautéed sample. The results showed that, after cooking, little if any dialyzable iron results from digestion of muscle proteins. Our research indicates that, in cooked chicken, residual acid-extractable components are the most important source of dialyzable iron. PMID:26904627

In situ biogas stripping of ammonia from a digester using a gas mixing system.

PubMed

Serna-Maza, Alba; Heaven, Sonia; Banks, Charles J

2017-12-01

Previous studies have suggested the use of digester biogas mixing systems for in situ ammonia removal from anaerobic digestates. The feasibility of this was tested at moderate and complete gas mixing rates at mesophilic and thermophilic temperatures in a 75-L digester. Experimental results showed that at gas mixing rates typical of full-scale commercial digesters the reduction in total ammonia nitrogen concentrations would be insufficient to allow stable acetoclastic methanogenesis in mesophilic conditions, or to prevent total inhibition of methanogenic activity in thermophilic food waste digestion. Simulation based on batch column stripping experiments at 55°C at gas violent flow rates of 0.032 m 3  m -2  min -1 indicated that ammonia concentrations could be reduced below inhibitory values in thermophilic food waste digestion for organic loading rates of up to 6 kg VS m -3  day -1 . These mixing rates are far in excess of those used in full-scale gas-mixed digesters and may not be operationally or commercially feasible.

Comparing environmental consequences of anaerobic mono- and co-digestion of pig manure to produce bio-energy--a life cycle perspective.

PubMed

De Vries, J W; Vinken, T M W J; Hamelin, L; De Boer, I J M

2012-12-01

The aim of this work was to assess the environmental consequences of anaerobic mono- and co-digestion of pig manure to produce bio-energy, from a life cycle perspective. This included assessing environmental impacts and land use change emissions (LUC) required to replace used co-substrates for anaerobic digestion. Environmental impact categories considered were climate change, terrestrial acidification, marine and freshwater eutrophication, particulate matter formation, land use, and fossil fuel depletion. Six scenarios were evaluated: mono-digestion of manure, co-digestion with: maize silage, maize silage and glycerin, beet tails, wheat yeast concentrate (WYC), and roadside grass. Mono-digestion reduced most impacts, but represented a limited source for bio-energy. Co-digestion with maize silage, beet tails, and WYC (competing with animal feed), and glycerin increased bio-energy production (up to 568%), but at expense of increasing climate change (through LUC), marine eutrophication, and land use. Co-digestion with wastes or residues like roadside grass gave the best environmental performance. Copyright © 2012 Elsevier Ltd. All rights reserved.

Protein enrichment of corn cob heteroxylan waste slurry by thermophilic aerobic digestion using Bacillus stearothermophilus.

PubMed

Ugwuanyi, J Obeta; Harvey, Linda M; McNeil, Brian

2008-10-01

Thermophilic aerobic digestion (TAD) of heteroxylan waste was implemented at waste load of 30gL(-1) with mineral nitrogen supplementation to study effect of the process on waste degradation, protein accretion and quality. Digestions were carried out at 45 50, 55, 60 and 65 degrees C using Bacillusstearothermophilus in a CSTR under batch conditions at 1.0vvm aeration rate, pH 7.0 for a maximum of 120h. Amylase and xylanase activities appeared rapidly in the digest, while basal protease activity appeared early in the digestion and increased towards end of the processes. Highest degradation of volatile suspended solid, hemicellulose and fibre occurred at 55 degrees C while highest degradation of total suspended solid occurred at 60 degrees C. Highest protein accretion (258.8%) and assimilation of mineral nitrogen and soluble protein occurred at 55 degrees C. The % content of amino acids of digest crude protein increased relative to raw waste and with digestion temperature. Quality of digest protein was comparable to the FAO standard for feed use. TAD has potentials for use in the protein enrichment of waste.

Acid digestion of geological and environmental samples using open-vessel focused microwave digestion.

PubMed

Taylor, Vivien F; Toms, Andrew; Longerich, Henry P

2002-01-01

The application of open vessel focused microwave acid digestion is described for the preparation of geological and environmental samples for analysis using inductively coupled plasma-mass spectrometry (ICP-MS). The method is compared to conventional closed-vessel high pressure methods which are limited in the use of HF to break down silicates. Open-vessel acid digestion more conveniently enables the use of HF to remove Si from geological and plant samples as volatile SiF4, as well as evaporation-to-dryness and sequential acid addition during the procedure. Rock reference materials (G-2 granite, MRG-1 gabbros, SY-2 syenite, JA-1 andesite, and JB-2 and SRM-688 basalts) and plant reference materials (BCR and IAEA lichens, peach leaves, apple leaves, Durham wheat flour, and pine needles) were digested with results comparable to conventional hotplate digestion. The microwave digestion method gave poor results for granitic samples containing refractory minerals, however fusion was the preferred method of preparation for these samples. Sample preparation time was reduced from several days, using conventional hotplate digestion method, to one hour per sample using our microwave method.

Methane production and characteristics of the microbial community in the co-digestion of spent mushroom substrate with dairy manure.

PubMed

Luo, Xiaosha; Yuan, Xufeng; Wang, Shiyu; Sun, Fanrong; Hou, Zhanshan; Hu, Qingxiu; Zhai, Limei; Cui, Zongjun; Zou, Yajie

2018-02-01

Spent mushroom substrate (SMS) is a potential biomass material generated during mushroom cultivation. In this study, the methane yield and microbial community resulting from co-digestion of SMS and dairy manure (DM) at different mixing ratios (0:4, 1:1, 3:1, and 1:3), were evaluated. Co-digestion analysis showed that the methane yield from the mixtures was 6%-61% higher than the yield from SMS or DM alone, indicating a synergistic effect of co-digestion of SMS with DM. For the SMS of F.velutipes (SFv) and P.erygii var. tuoliensis (SPt), co-digestion of DM/SMS at a ratio of 1:1 was optimal, but for the SMS of P. eryngi (SPe), co-digestion of DM/SMS at a ratio of 3:1 was ideal. The pH at all co-digestion ratios was in the range of 6.8-8.0, indicating that adding DM could increase the systemic buffering capacity. Methanosaetaceae was shown to be the predominant methanogens present during the co-digestion of DM/SMS. Copyright © 2017 Elsevier Ltd. All rights reserved.

Application of Pineapple Juice in the Fish Digestion Process for Carcinogenic Liver Fluke Metacercaria Collection

PubMed Central

Sripan, Panupan; Aukkanimart, Ratchadawan; Boonmars, Thidarut; Pranee, Sriraj; Songsri, Jiraporn; Boueroy, Parichart; Khueangchaingkhwang, Sukhonthip; Pumhirunroj, Benjamabhorn; Artchayasawat, Atchara

2017-01-01

Pepsin is common digestive enzyme used for fish digestion in the laboratory to collect trematode metacercariae. In a field study, to survey the infected fish is needed a huge yield of pepsin and it is very expensive. Therefore, our purpose of this study was to investigate the candidate enzyme from pineapple juice which has a digestive enzyme called bromelain, a mixture of proteolytic enzymes, to digest fish in order to harvest metacercariae. Fish were divided into 2 groups: one group in which metacercariae were harvested using acid pepsin as a control and other groups in which the fish was digested using fresh pineapple juices. The results showed that pineapple juice is able to digest fish similarly to pepsin. The Pattavia pineapple juice had the highest number of metacercariae similar to the control. For Trat Si Thong pineapple juice, we found the number of metacercariae was less than control. This result suggests that the Pattavia pineapple juice was optimal juice for fish digestion to metacercaria collection and can be used instread of pepsin acid. PMID:28441786

Effect of heat–moisture treatment on digestibility of different cultivars of sweet potato (Ipomea batatas (L.) Lam) starch

PubMed Central

Senanayake, Suraji; Gunaratne, Anil; Ranaweera, K K D S; Bamunuarachchi, Arthur

2014-01-01

Different heat–moisture levels were applied to native starches from different cultivars of sweet potatoes available in Sri Lanka (Wariyapola red, Wariyapola white, Pallepola variety, Malaysian variety and CARI 273) to study the digestibility level. Samples were treated with 20, 25, and 30% moisture at 85°C and 120°C for 6 h and in vitro starch digestibility was tested with porcine pancreatin enzyme. A range of 19.3–23.5% digestibility was shown by the native starches with no significant difference (P < 0.05). Significant changes were observed in the digestibility level of the hydrothermally modified starches and the moisture content showed a positive impact on the digestibility. Heat–moisture treatment at 85°C brought an overall increase in digestibility and temperature beyond 85°C had a negative impact. No significant difference (P < 0.05) in the digestibility was observed with 20% and 25% moisture at 85°C and increased level were seen at 85°C and 30% moisture. PMID:25473497

Variation in cell wall composition among forage maize (Zea mays L.) inbred lines and its impact on digestibility: analysis of neutral detergent fiber composition by pyrolysis-gas chromatography-mass spectrometry.

PubMed

Fontaine, Anne-Sophie; Bout, Siobhán; Barrière, Yves; Vermerris, Wilfred

2003-12-31

Cell wall digestibility is an important determinant of forage quality, but the relationship between cell wall composition and digestibility is poorly understood. We analyzed the neutral detergent fiber (NDF) fraction of nine maize inbred lines and one brown midrib3 mutant with pyrolysis-gas chromatography-mass spectrometry (Py-GC-MS). Among 29 pyrolysis fragments that were quantified, two carbohydrate-derived and six lignin-derived fragments showed statistically significant genetic variation. The pyrolysis products 4-vinyl phenol and 2,6-dimethoxy-4-vinyl phenol were negatively correlated with digestibility, whereas furfural and 3-(4-hydroxyphenyl)-3-oxopropanal showed a positive correlation with digestibility. Linear discriminant analysis of the pyrolysis data resulted in the resolution of groups of inbred lines with different digestibility properties based on their chemical composition. These analyses reveal that digestibility is governed by complex interactions between different cell wall compounds, but that several pyrolysis fragments can be used as markers to distinguish between maize lines with different digestibility.

Cooking Chicken Breast Reduces Dialyzable Iron Resulting from Digestion of Muscle Proteins.

PubMed

Gokhale, Aditya S; Mahoney, Raymond R

2014-01-01

The purpose of this research was to study the effect of cooking chicken breast on the production of dialyzable iron (an in vitro indicator of bioavailable iron) from added ferric iron. Chicken breast muscle was cooked by boiling, baking, sautéing, or deep-frying. Cooked samples were mixed with ferric iron and either extracted with acid or digested with pepsin and pancreatin. Total and ferrous dialyzable iron was measured after extraction or digestion and compared to raw chicken samples. For uncooked samples, dialyzable iron was significantly enhanced after both extraction and digestion. All cooking methods led to markedly reduced levels of dialyzable iron both by extraction and digestion. In most cooked, digested samples dialyzable iron was no greater than the iron-only (no sample) control. Cooked samples showed lower levels of histidine and sulfhydryls but protein digestibility was not reduced, except for the sautéed sample. The results showed that, after cooking, little if any dialyzable iron results from digestion of muscle proteins. Our research indicates that, in cooked chicken, residual acid-extractable components are the most important source of dialyzable iron.

Biogas by semi-continuous anaerobic digestion of food waste.

PubMed

Zhang, Cunsheng; Su, Haijia; Wang, Zhenbin; Tan, Tianwei; Qin, Peiyong

2015-04-01

The semi-continuous anaerobic digestion of food waste was investigated in 1-L and 20-L continuously stirred tank reactors (CSTRs), to identify the optimum operation condition and the methane production of the semi-continuous anaerobic process. Results from a 1-L digester indicated that the optimum organic loading rate (OLR) for semi-continuous digestion is 8 g VS/L/day. The corresponding methane yield and chemical oxygen demand (COD) reduction were 385 mL/g VS and 80.2 %, respectively. Anaerobic digestion was inhibited at high OLRs (12 and 16 g VS/L/day), due to volatile fatty acid (VFA) accumulation. Results from a 20-L digester indicated that a higher methane yield of 423 mL/g VS was obtained at this larger scale. The analysis showed that the methane production at the optimum OLR fitted well with the determined kinetics equation. An obvious decrease on the methane content was observed at the initial of digestion. The increased metabolization of microbes and the activity decrease of methanogen caused by VFA accumulation explained the lower methane content at the initial of digestion.

Mechanisms of starch digestion by α-amylase-Structural basis for kinetic properties.

PubMed

Dhital, Sushil; Warren, Frederick J; Butterworth, Peter J; Ellis, Peter R; Gidley, Michael J

2017-03-24

Recent studies of the mechanisms determining the rate and extent of starch digestion by α-amylase are reviewed in the light of current widely-used classifications for (a) the proportions of rapidly-digestible (RDS), slowly-digestible (SDS), and resistant starch (RS) based on in vitro digestibility, and (b) the types of resistant starch (RS 1,2,3,4…) based on physical and/or chemical form. Based on methodological advances and new mechanistic insights, it is proposed that both classification systems should be modified. Kinetic analysis of digestion profiles provides a robust set of parameters that should replace the classification of starch as a combination of RDS, SDS, and RS from a single enzyme digestion experiment. This should involve determination of the minimum number of kinetic processes needed to describe the full digestion profile, together with the proportion of starch involved in each process, and the kinetic properties of each process. The current classification of resistant starch types as RS1,2,3,4 should be replaced by one which recognizes the essential kinetic nature of RS (enzyme digestion rate vs. small intestinal passage rate), and that there are two fundamental origins for resistance based on (i) rate-determining access/binding of enzyme to substrate and (ii) rate-determining conversion of substrate to product once bound.

Lyophilization decreases the formation of dialyzable iron by extraction and digestion of chicken breast muscle.

PubMed

Karava, Nilesh B; Mahoney, Raymond R

2011-06-01

We studied the effect of lyophilization of chicken breast muscle on the formation of dialyzable iron from ferric iron. Chicken breast muscle was used chilled, frozen or lyophilized and was analyzed for sulfhydryl and histidine content. It was then homogenized and mixed with ferric iron. The mixture was extracted with acid or digested with pepsin and pancreatin. The extracts and digests were analyzed for dialyzable ferrous and dialyzable total iron and also for protein. In the chilled muscle, similar amounts of dialyzable iron were formed after acid extraction and after proteolytic digestion; however, digestion led to more dialyzable ferrous iron. Freezing had no effect but lyophilization of the homogenized muscle caused large decreases in dialyzable iron and dialyzable ferrous iron for both extraction and digestion processes. Lyophilization also resulted in decreased extraction of peptides, decreased digestion of muscle proteins and reduced levels of sulfhydryl and histidine residues. Our results demonstrate that dialyzable iron is produced both by acid-soluble low molecular weight muscle component(s) and also by peptides resulting from digestion of muscle proteins: both of which reduce and chelate iron. Reduced formation of dialyzable iron by both mechanisms following lyophilization could be explained by sulfhydryl oxidation and impaired digestion due to protein crosslinking.

Short communication: Intestinal digestibility of amino acids in fluid- and particle-associated rumen bacteria determined using a precision-fed cecectomized rooster bioassay.

PubMed

Fonseca, A C; Fredin, S M; Ferraretto, L F; Parsons, C M; Utterback, P L; Shaver, R D

2014-01-01

Microbial protein represents the majority of metabolizable protein absorbed by ruminant animals. Enhanced understanding of the AA digestibility of rumen microbes will improve estimates of metabolizable protein. The objective of this experiment was to determine the digestibility of AA in fluid- (FAB) and particle-associated bacteria (PAB) using the precision-fed cecectomized rooster bioassay. Bacteria were isolated from 4 ruminally cannulated lactating Holstein cows by differential centrifugation, including particle suspension in 0.1% Tween-80 for increased removal of PAB from ruminal digesta. Samples of FAB and PAB were fed to 9 cecectomized roosters to determine standardized digestibility of AA. Total AA digestibility was 76.8 and 75.5% for FAB and PAB, respectively, but did not differ. Differences existed in AA digestibilities within bacterial type when compared with the mean essential AA digestibility value. Compared with previous literature estimates of AA digestibility in microbes (mean = 76%; range = 57-87%) and relative to National Research Council estimates of total AA from rumen bacteria (80%), the precision-fed cecectomized rooster assay is an acceptable in vivo model to determine AA digestibility of rumen bacteria. Copyright © 2014 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Long-term thermophilic mono-digestion of rendering wastes and co-digestion with potato pulp.

PubMed

Bayr, S; Ojanperä, M; Kaparaju, P; Rintala, J

2014-10-01

In this study, mono-digestion of rendering wastes and co-digestion of rendering wastes with potato pulp were studied for the first time in continuous stirred tank reactor (CSTR) experiments at 55°C. Rendering wastes have high protein and lipid contents and are considered good substrates for methane production. However, accumulation of digestion intermediate products viz., volatile fatty acids (VFAs), long chain fatty acids (LCFAs) and ammonia nitrogen (NH4-N and/or free NH3) can cause process imbalance during the digestion. Mono-digestion of rendering wastes at an organic loading rate (OLR) of 1.5 kg volatile solids (VS)/m(3)d and hydraulic retention time (HRT) of 50 d was unstable and resulted in methane yields of 450 dm(3)/kg VS(fed). On the other hand, co-digestion of rendering wastes with potato pulp (60% wet weight, WW) at the same OLR and HRT improved the process stability and increased methane yields (500-680 dm(3)/kg VS(fed)). Thus, it can be concluded that co-digestion of rendering wastes with potato pulp could improve the process stability and methane yields from these difficult to treat industrial waste materials. Copyright © 2014 Elsevier Ltd. All rights reserved.

Food waste co-digestion with slaughterhouse waste and sewage sludge: Digestate conditioning and supernatant quality.

PubMed

Borowski, Sebastian; Boniecki, Paweł; Kubacki, Przemysław; Czyżowska, Agata

2018-04-01

In this study, the anaerobic mesophilic co-digestion of food waste (FW) with municipal sewage sludge (MSS) and slaughterhouse waste (SHW) was undertaken in 3-dm 3 laboratory reactors as well as in 50-dm 3 reactors operated in semi-continuous conditions. The highest methane yield of around 0.63 m 3 CH 4 /kgVS fed was achieved for the mixture of FW and SHW treated in the laboratory digester operated at solids retention time (SRT) of 30 days, whereas the co-digestion of FW with MSS under similar operating conditions produced 0.46 m 3 of methane from 1 kgVS fed . No significant differences between methane yields from laboratory digesters and large-scale reactors were reported. The conditioning tests with the digestates from reactor experiments revealed the highest efficiency of inorganic coagulants among all investigated chemicals, which applied in a dose of 10 g/kg allowed to reduce capiliary suction time (CST) of the digestate below 20 s. The combined conditioning with coagulants and bentonite did not further reduce the CST value but improved the quality of the digestate supernatant. In particular, the concentrations of suspended solids, COD as well as metals in the supernatant were considerably lowered. Copyright © 2017. Published by Elsevier Ltd.

A new in vitro lipid digestion - in vivo absorption model to evaluate the mechanisms of drug absorption from lipid-based formulations.

PubMed

Crum, Matthew F; Trevaskis, Natalie L; Williams, Hywel D; Pouton, Colin W; Porter, Christopher J H

2016-04-01

In vitro lipid digestion models are commonly used to screen lipid-based formulations (LBF), but in vitro-in vivo correlations are in some cases unsuccessful. Here we enhance the scope of the lipid digestion test by incorporating an absorption 'sink' into the experimental model. An in vitro model of lipid digestion was coupled directly to a single pass in situ intestinal perfusion experiment in an anaesthetised rat. The model allowed simultaneous real-time analysis of the digestion and absorption of LBFs of fenofibrate and was employed to evaluate the influence of formulation digestion, supersaturation and precipitation on drug absorption. Formulations containing higher quantities of co-solvent and surfactant resulted in higher supersaturation and more rapid drug precipitation in vitro when compared to those containing higher quantities of lipid. In contrast, when the same formulations were examined using the coupled in vitro lipid digestion - in vivo absorption model, drug flux into the mesenteric vein was similar regardless of in vitro formulation performance. For some drugs, simple in vitro lipid digestion models may underestimate the potential for absorption from LBFs. Consistent with recent in vivo studies, drug absorption for rapidly absorbed drugs such as fenofibrate may occur even when drug precipitation is apparent during in vitro digestion.

Effect of endogenous proteins and lipids on starch digestibility in rice flour.

PubMed

Ye, Jiangping; Hu, Xiuting; Luo, Shunjing; McClements, David Julian; Liang, Lu; Liu, Chengmei

2018-04-01

The composition and structure of the food matrix can have a major impact on the digestion. The aim of this work was to investigate the effects of endogenous proteins and lipids on starch digestibility in rice flour, with an emphasis on establishing the underlying physicochemical mechanisms involved. Native long-grain indica rice flour and rice flour with the lipids and/or proteins removed were subjected to a simulated digestion in vitro. A significant increase in starch digestibility was observed after removal of proteins, lipids, or both. The starch digestibility of the rice flour without lipids was slightly lower than that without proteins, even though the proteins content was about 10-fold higher than the lipids content. Microstructural analysis suggested that the proteins and lipids were normally attached to the surfaces of the starch granules in the native rice flour, thus inhibiting their contact with digestive enzymes. Moreover, the proteins and lipids restricted the swelling of the starch granules, which may have decreased their digestion by reducing their surface areas. In addition, amylose-lipid complex was detected in the rice flour, which is also known to slow down starch digestion. These results have important implications for the design of foods with improved nutritional profiles. Copyright © 2018 Elsevier Ltd. All rights reserved.

Effects of trypsinization and of a combined trypsin, collagenase, and DNase digestion on liberation and in vitro function of satellite cells isolated from juvenile porcine muscles.

PubMed

Miersch, Claudia; Stange, Katja; Röntgen, Monika

2018-06-01

Muscle stem cells, termed satellite cells (SC), and SC-derived myogenic progenitor cells (MPC) are involved in postnatal muscle growth, regeneration, and muscle adaptability. They can be released from their natural environment by mechanical disruption and tissue digestion. The literature contains several isolation protocols for porcine SC/MPC including various digestion procedures, but comparative studies are missing. In this report, classic trypsinization and a more complex trypsin, collagenase, and DNase (TCD) digestion were performed with skeletal muscle tissue from 4- to 5-d-old piglets. The two digestion procedures were compared regarding cell yield, viability, myogenic purity, and in vitro cell function. The TCD digestion tended to result in higher cell yields than digestion with solely trypsin (statistical trend p = 0.096), whereas cell size and viability did not differ. Isolated myogenic cells from both digestion procedures showed comparable proliferation rates, expressed the myogenic marker Desmin, and initiated myogenic differentiation in vitro at similar levels. Thus, TCD digestion tended to liberate slightly more cells without changes in the tested in vitro properties of the isolated cells. Both procedures are adequate for the isolation of SC/MPC from juvenile porcine muscles but the developmental state of the animal should always be considered.

The fate and effect of monensin during anaerobic digestion of dairy manure under mesophilic conditions

PubMed Central

Mulbry, Walter; Rice, Clifford; Lansing, Stephanie

2018-01-01

There is growing concern about residual antibiotics and feed additives in the manure of treated animals because of the effects of these residues in the environment. Monensin is the most widely used ionophore coccidiostat in the U.S. The objective of this study was to determine the fate and effect of monensin during the anaerobic digestion of dairy manure. Duplicate plug flow field-scale digesters were operated using non-amended dairy manure and dairy manure amended with monensin to 1 and 10 mg/L for 56 days at 30°C at an organic loading rate of 1.4 kg VS/m3-d and 17-day hydraulic retention time. Results showed that monensin was reduced approximately 70% during anaerobic digestion. Methane production from digesters using manure amended with 1 mg/L monensin was comparable to that from digesters operated without added monensin. However, digesters using manure amended with 10 mg/L monensin yielded 75% less methane than digesters using manure without added monensin. These results suggest that anaerobic digestion is an effective treatment for reducing, but not eliminating, monensin in dairy manure. Monensin did not reduce methane production at concentrations expected in dairy manure at recommended dosage rates. PMID:29420605

Nitrous oxide and methane emissions during storage of dewatered digested sewage sludge.

PubMed

Willén, Agnes; Rodhe, Lena; Pell, Mikael; Jönsson, Håkan

2016-12-15

This study investigated the effect on greenhouse gas emissions during storage of digested sewage sludge by using a cover during storage or applying sanitisation measures such as thermophilic digestion or ammonia addition. In a pilot-scale storage facility, nitrous oxide and methane emissions were measured on average twice monthly for a year, using a closed chamber technique. The thermophilically digested sewage sludge (TC) had the highest cumulative emissions of nitrous oxide (1.30% of initial total N) followed by mesophilically digested sewage sludge stored without a cover (M) (0.34%) and mesophilically digested sewage sludge stored with a cover (MC) (0.19%). The mesophilically digested sewage sludge sanitised with ammonia and stored with a cover (MAC) showed negligible cumulative emissions of nitrous oxide. Emissions of methane were much lower from TC and MAC than from M and MC. These results indicate that sanitisation by ammonia treatment eliminates the production of nitrous oxide and reduces methane emissions from stored sewage sludge, and that thermophilic digestion has the potential to reduce the production of methane during storage compared with mesophilic digestion. The results also indicate a tendency for lower emissions of nitrous oxide and higher emissions of methane from covered sewage sludge compared with non-covered. Copyright © 2016 Elsevier Ltd. All rights reserved.