Rapid Method for the Determination of Diaminopimelic Acid Using Ion Exchange Column Chromatography

Csapó, J.; Csapó-Kiss, Z.; Csordás, E.; Martín, T. G.; Folestad, Staffan; Tivesten, Anna; Nemethy, Sandor

doi:10.1080/00032719508000024

Cited by 6 publications

(6 citation statements)

References 8 publications

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“…This method is based on the estimation of the ratio diaminopimelic acid/nitrogen in ruminal bacteria and the amount of the microbial marker in digesta (5) . From these values the amount of bacterial nitrogen in intestinal digesta is calculated (68) . Briefly, lyophilized samples are hydrolyzed with methasulfonic acid then centrifuged.…”

Section: The Use Of Daminopimelic Acid As a Microbial Markermentioning

confidence: 99%

“…Diaminopimelic acid is found in the cell membrane of ruminal bacteria and it is absent in feedstuffs commonly fed to ruminants (68) . The accuracy of the technique depends on a constant diaminopimelic acid/nitrogen ratio among various microbial species, or the maintenance of a constant ratio of microbial species in the rumen.…”

Section: The Use Of Daminopimelic Acid As a Microbial Markermentioning

confidence: 99%

“…In addition, peptidoglycan concentration varies among bacteria, thus they have different diaminopimelic acid concentrations. Gram-positive bacteria contain more peptidoglycan in the cell wall than gram-negative bacteria (68) . Given the greater proportion of Firmicutes (phylum), Clostridiales and Coriobacteriales (orders) and Lachnospiraceae (family), largely represented by gram-positive bacteria, in the reference pellets isolated from ruminal fluid compared to bacteria from whole intestinal digesta, when diaminopimelic acid is used as microbial marker, researchers should be aware of potential underestimation of intestinal microbial protein flow.…”

Section: Implications Of Taxonomic Differences Between Bacteria From mentioning

confidence: 99%

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Factors affecting the ruminal microbial composition and methods to determine microbial protein yield. Review

Castillo‐Lopez¹,

Dominguez²

2019

Rev. Mex. Cienc. Pecu.

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Microbial protein synthesized in the rumen is a major contributor of metabolizable protein. Thus, accurate estimation of microbial protein is essential in ruminant nutrition. The objective of this review is to describe the microbial composition, major factors affecting its yield and methods to estimate microbial protein flow to the intestine. The use of novel molecular techniques to elucidate the ruminal microbiome and improve methods for estimating microbial protein are discussed. Bacteria, protozoa, fungi and archaea compose the ruminal microbiome. Main factors affecting microbial protein synthesis are availability of carbohydrates, ruminally degradable protein, dietary fat, and ruminal pH. Major microbial markers used to estimate microbial protein synthesis are total purines, diaminopimelic acid and labeled nitrogen; in addition, DNA through real-time PCR is being tested for the estimation of bacterial, protozoal and yeast protein separately. The main difficulty in the estimation of microbial protein flow is obtaining representative microbial pellets from the rumen, which are used as reference to establish the ratio of marker/nitrogen. Detailed phylogenetic analysis using High-throughput DNA sequencing has recently revealed drastic Rev Mex Cienc Pecu 2019;10(1):120-148 121 taxonomic differences between fluid-associated bacteria and bacteria from whole intestinal digesta contents. For example, ruminal fluid contains less Fibrobacteres and Proteobacteria, but more Firmicutes compared to whole intestinal digesta. This demonstrates the need of developing effective bacterial collection procedures for obtaining representative ruminal microbial reference pellets to prevent bias on the estimation of the contribution of microbial protein to the intestinal supply of metabolizable protein.

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Section: The Use Of Daminopimelic Acid As a Microbial Markermentioning

confidence: 99%

Section: The Use Of Daminopimelic Acid As a Microbial Markermentioning

confidence: 99%

Section: Implications Of Taxonomic Differences Between Bacteria From mentioning

confidence: 99%

See 1 more Smart Citation

Factors affecting the ruminal microbial composition and methods to determine microbial protein yield. Review

Castillo‐Lopez¹,

Dominguez²

2019

Rev. Mex. Cienc. Pecu.

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“…Estimation of Bacterial CP using Diaminopumelic Acid as a Bacterial Marker. The use of DAPA as a bacterial marker has been widely reported (Dufva et al, 1982;Webster et al, 1990;Csapo et al, 2008). In the present study, isolated ruminal bacterial pellets were collected and duodenal digesta samples were analyzed for DAPA using a 717 HPLC system (Waters Corporation Inc., Milford, MA) according to the procedure described by Webster et al (1990).…”

Section: Analysis Of Feed Duodenal Digesta and Bacterialmentioning

confidence: 99%

In vivo determination of rumen undegradable protein of dried distillers grains with solubles and evaluation of duodenal microbial crude protein flow

Castillo‐Lopez

Klopfenstein

Fernando

et al. 2013

Journal of Animal Science

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The objectives of this trial were to determine the rumen undegradable protein (RUP) of dried distillers grains with solubles (DDGS), to compare the estimates of duodenal bacterial CP (BCP) flow using diaminopumelic acid (DAPA) or DNA as bacterial markers, and to estimate duodenal protozoal CP (PCP) and yeast CP (YCP) flow when DDGS are fed. Three crossbred steers fitted with ruminal and double L-shaped duodenal cannulae (average BW 780 ± 137 kg) were used in a 3 treatment, 6 period crossover design. Animals were housed in individual free stalls and fed twice daily at 0700 and 1900 h. Diets (DM basis) were 1) CONTROL, which is 0% DDGS but with 19.5% corn bran, 20% sorghum silage, 60% brome hay, 0.5% trace minerals, and 0.25% urea, 2) LOW DDGS, which is inclusion of 9.75% DDGS replacing equal percentage of corn bran, and 3) HIGH DDGS, which is inclusion of 19.5% DDGS completely replacing corn bran. Duodenal BCP flow was estimated using DAPA and DNA as bacterial markers. In addition, duodenal PCP and YCP flow were estimated using DNA markers. The value of DDGS RUP as a percent of CP was determined to be 63.0 ± 0.64%. Estimates of duodenal BCP flow using DAPA were 473, 393, and 357 ± 78 g/d (P = 0.09) for CONTROL, LOW DDGS, and HIGH DDGS, respectively. Estimates of duodenal BCP flow using DNA were 479, 397, and 368 ± 74 g/d (P = 0.14), respectively. Average BCP flow across treatments was unaffected (P = 0.71) by marker type and were 404 and 417 ± 83 g/d for DAPA and DNA markers, respectively. Estimates of duodenal PCP flow were 82, 80, and 78 ± 12 g/d (P = 0.64) for CONTROL, LOW DDGS, and HIGH DDGS, respectively. Estimates of duodenal YCP flow were 0.15, 1.94, and 4.80 ± 0.66 g/d (P < 0.01) for CONTROL, LOW DDGS, and HIGH DDGS, respectively. Duodenal BCP flow tended to decrease with DDGS inclusion, but estimates were not affected by marker type. In addition, DDGS did not affect duodenal PCP supply and provided small amounts of duodenal YCP. Overall, the value of DDGS RUP determined in this study will contribute to a better understanding of the effect of this coproduct in ruminant nutrition.

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“…We have used performic acid oxidation preceding hydrolysis of proteins for AA analysis. Although oxidative hydrolysis was proved to not interfere in the determination of DAPA (Csapó et al, 1995), we observed that the appearance of DAPA peaks on the chromatogram was not sharp and well defined. We hypothesized that during oxidative hydrolysis, new substances originated that coeluted with DAPA, giving no reliable measurements, and this may be a particular problem for this group of feedstuff.…”

Section: Discussionmentioning

confidence: 85%

In situ ruminal degradation of amino acids and in vitro protein digestibility of undegraded CP of dried distillers’ grains with solubles from European ethanol plants

Westreicher-Kristen

Steingaß

Rodehutscord

2013

Animal

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The objectives of this study were to compare the in situ ruminal degradation of CP and amino acids (AAs) of dried distillers' grains with solubles (DDGS), and to estimate intestinal digestibility (ID) of undegradable crude protein (UDP) with the in vitro pepsinpancreatin solubility of CP (PPS), using either DDGS samples (DDGS-s) or DDGS residues (DDGS-r) obtained after 16 h ruminal incubation. Thirteen samples originating from wheat, corn, barley and blends were studied. Lysine and methionine content of DDGS-s varied from 1.4 to 4.0 and 1.3 to 2.0 g/16 g N, respectively. The milk protein score (MPS) of DDGS-s was low and ranged from 0.36 to 0.51, and lysine and isoleucine were estimated to be the most limiting AAs in DDGS-s and DDGS-r. DDGS-r contained slightly more essential AAs (EAAs) than did the DDGS-s. Rumen degradation after 16 h varied from 44% to 94% for CP, from 39% to 90% for lysine and from 35% to 92% for methionine. Linear regressions showed that the ruminal degradation of individual AAs can be predicted from CP degradation. The PPS of DDGS-s was higher than that of DDGS-r and it varied from 70% to 89% and from 47% to 81%, respectively. There was no significant correlation between the PPS of DDGS-s and PPS of DDGS-r ( R 2 = 0.31). The estimated intestinally absorbable dietary protein (IADP) averaged 21%. Moderate correlation was found between the crude fibre (CF) content and PPS of DDGS-r ( R 2 = 0.43). This study suggests an overestimation of the contribution of UDP of DDGS to digestible protein supply in the duodenum in some currently used protein evaluation systems. More research is required and recommended to assess the intestinal digestibility of AAs from DDGS.

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Rapid Method for the Determination of Diaminopimelic Acid Using Ion Exchange Column Chromatography

Cited by 6 publications

References 8 publications

Factors affecting the ruminal microbial composition and methods to determine microbial protein yield. Review

Factors affecting the ruminal microbial composition and methods to determine microbial protein yield. Review

In vivo determination of rumen undegradable protein of dried distillers grains with solubles and evaluation of duodenal microbial crude protein flow

In situ ruminal degradation of amino acids and in vitro protein digestibility of undegraded CP of dried distillers’ grains with solubles from European ethanol plants

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