Bacterial D-alanine concentrations as a marker of bacterial nitrogen in the gastrointestinal tract of pigs and cows

Schoenhusen, U.; Voigt, J.; Hennig, U.; Kuhla, S.; Žitňan, R.; Souffrant, W. B.

doi:10.17221/1922-vetmed

Cited by 8 publications

(8 citation statements)

References 34 publications

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“…It is a well known peptidoglycan amino acid bound to bacterial cell walls of gram-negative bacteria (Ling 1990). Gram-positive bacteria contain approximately 30-60% peptidoglycan, whereas gram-negative bacteria have only 10% peptidoglycan (Schoenhusen et al 2008). Amino acid composition and structure of peptidoglycans in gram-positive bacteria varies greatly, whereas that of gram-negative bacteria is more consistent (Miner-Williams et al 2013).…”

mentioning

confidence: 99%

2,6-Diaminopimelic acid as a biological marker of rumen synthesis and fermentation capacities in the transition period and early lactation of dairy cows

Maskaľová¹,

Vajda²,

Bujňák³

2014

Acta Vet. Brno

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The experiment was conducted to evaluate the effect of different types of available nutrients of total mix rations on fermentation and synthesis capacities of the rumen in Holstein dairy cows divided into groups in the transition period (± 3 weeks after parturition) and early lactation (60-80 days in milk). The diets were fed as a total mixed ration. Results obtained with feeding Diet C (high concentration of available nutrients) to dairy cows confirmed a significant increase in 2,6-diaminopimelic acid from the initial mean 2.2 μg/ml at 21 days before calving to 6.2 μg/ml (P < 0.01) at 21 days after calving or 7.6 μg/ml (P < 0.01) at early lactation, and a parallel significant increase in the concentration of volatile fatty acids (P < 0.01). The concentration of propionic acid increased from the initial mean 25.3 to 39.3 mmol/l (P < 0.01) in the transition period, and to 43.1 mmol/l at early lactation (P < 0.01). All indicators of rumen fermentation and synthesis determined in dairy cows with Diet B (low concentration of available nutrients) of the total mix ration according to productive phases were not significant. High regression coefficients of examined indicators of 2,6-diaminopimelic acid to volatile fatty acid (r = 0.813), propionic acid (r = 0.780) and acetic acid (r = 0.635) indicate direct correlation between fermentation and synthesis activities of the rumen. Our results show that 2,6-diaminopimelic acid appears as a perspective marker for the monitoring and evaluation of the synthesis function of the rumen and production of bacterial biomass under breeding conditions. VFA, DAPA, feeding and nutrition, microflora, fermentationRumen fermentation of the feed's organic matter and microbial biomass synthesis are reported to have the potential to meet 70-85% of the energy requirements and 70-100% of the protein needs of ruminants, even at higher levels of production. Microbial proteins synthesized in the rumen cover 80% of daily amino acid needs and constitute 50-80% of total absorbable amino acids (Bach et al. 2005). Synthesis of microbial proteins is affected by usability of fermentable carbohydrates (Ahvenjarvi et al. 2002), nitrogen (Verbic 2002) and minerals (Broudiscou et al. 1999). Optimum production of rumen microflora and synthesis of microbial proteins at the level of 12.6 g/100 g fermentable organic matter is ensured when the concentration of NH 3 in the ruminal fluid varies in the range 8.8-14.7 mmol/l which is an accepted reference value for assessment of the level of protein saturation in ruminant rations (Vajda et al. 1994; NRC 2001). Microbial efficiency expressed as g of microbial nitrogen per kg of dry matter, organic matter, total carbohydrate, and 100 g N digested, ranged from 20.8 to 26.6; 34.7 to 44.3; 35.1 to 50.3 and 89 to 95 g, respectively (Sniffen et al. 2006). Multiplication and growth of rumen microflora is affected by nutritional (quantity and ratio of carbohydrates and proteins) and dietetic factors (structure of total mix ration, dry matter intake, production of sal...

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confidence: 99%

2,6-Diaminopimelic acid as a biological marker of rumen synthesis and fermentation capacities in the transition period and early lactation of dairy cows

Maskaľová¹,

Vajda²,

Bujňák³

2014

Acta Vet. Brno

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“…D-alanine, serving as a marker for microbial nitrogen, was determined in ileal digesta according to the method of Schoenhusen et al (2008).…”

Section: Methodsmentioning

confidence: 99%

“…At present, the use of D-alanine as a bacterial marker is prevalent in the ruminant studies in which it is regarded as superior to DAPA (Garrett et al, 1987;Schoenhusen et al, 2008). There is only one short communication that refers to the use of D-alanine in pigs (Hennig et al, 1999).…”

Section: Days Post Dietary Change Total Ileal N Excretion (G/d)mentioning

confidence: 99%

Impact of diet composition on ileal digestibility and small intestinal morphology in early-weaned pigs fitted with a T-cannula

Kluess

Schoenhusen

Souffrant

et al. 2010

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Piglets, separated from their dam at 12 days of age and fed a milk substitute hourly, were used as a model for suckling. Animals were fitted with a terminal ileal T-cannula and a jugular vein catheter. At 28 days of age, half of the pigs had a dietary change to a cereal-based weaner diet fed as slurry, and the others remained on milk substitute. Animals were labelled by oral administration of 15 N-labelled yeast for 10 days (days 15 to 25). Blood samples were taken twice a day to monitor 15 N enrichment of the blood plasma. Diets included polyethylenglycol (PEG 4000) to allow calculation of apparent ileal digestibility of nitrogen and individual amino acids. Ileal bacterial nitrogen was calculated from D-alanine content of the digesta. Furthermore, small intestinal (SI) villus height and crypt depth were measured. Feed intake was increased by the dietary change. The total nitrogen flow was 3.2 6 0.4 g/day and 5.9 6 0.4 for the milk and weaner diet, respectively. Endogenous nitrogen flow at the terminal ileum was similar for both groups (milk diet 2.4 6 0.4 v. weaner diet 2.2 6 0.3 g/day), whereas the bacterial nitrogen content (0.08 6 0.01 g/day milk diet v. 0.15 6 0.01 g/day weaner diet, P , 0.01) and exogenous nitrogen flow (0.94 6 0.16 g/day milk diet v. 3.29 6 0.12 g/day weaner diet, P , 0.001) increased significantly in the weaner-diet group. The ileal apparent digestibility coefficient of protein was 0.81 6 0.06 and 0.68 6 0.01 for the milk replacer and the weaner diet, respectively. Morphology measurements made along the SI at 25%, 50% and 75% were similar between piglets fed milk replacer and those fed a cereal-based weaner diet. The only statistical effect ( P , 0.01) of dietary change was an increase in crypt depth in the weaner-diet group. In conclusion, pigs, following a dietary change analogous to weaning, lack the capacity to fully digest a standard weaner diet. This may result in an increased nutrient content entering the large intestine and an altered microbiota. In the absence of a period of anorexia, often associated with traditional weaning, we saw no evidence of villous atrophy, but report here a significant crypt hyperplasia, especially at the 75% level, as a result of dietary change.

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“…However, the peptidoglycan content of bacterial cell walls varies according to the cell type. Gram‐positive bacteria contain approximately 30–60% peptidoglycan, whereas gram‐negative bacteria have only 10% peptidoglycan (Schoenhusen et al., 2008). The amino acid composition and structure of peptidoglycans in gram‐positive bacteria varies greatly, whereas that of gram‐negative bacteria is more consistent.…”

Section: Introductionmentioning

confidence: 99%

“…However, the suitability of these markers is subject to debate. While some researchers show that d ‐Ala is an effective bacterial marker, (Garrett et al., 1987) others claim the reverse (Schoenhusen et al., 2008).…”

Section: Introductionmentioning

confidence: 99%

Comparison of three markers for the determination of bacterial protein in terminal ileal digesta in the growing pig

Miner-Williams

Moughan

Fuller

2012

Animal Physiology Nutrition

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The aim of the study was to compare three methods commonly used to determine the concentrations of bacterial protein in digesta collected from the terminal ileum of growing pigs that had been fed a casein-based diet. The amounts of bacterial protein in terminal ileal digesta were determined using three different markers: 2.6-diaminopimelic acid (DAPA) and the d-amino acids, d-aspartic acid (d-Asp) and d-glutamic acid (d-Glu). The effectiveness of each marker was compared against a control based on physical fractionation by centrifugation. The total bacterial protein concentrations derived from the markers d-Asp and d-Glu were significantly different (p = 0.05) to those calculated from DAPA and the control, but there was no difference between DAPA and the control. The percentage of bacterial nitrogen ranged from 40% to 52% dependent on the marker used. Bacterial protein expressed as a percentage of the total protein, ranged from 48% to 62%, a substantial proportion of which (12-28%) was derived from lysed bacterial cells. Statistical correlations between the estimation methods were low. Such poor correlation between the markers may be the result of random errors such as variance in the epimerization of the two d-amino acids during protein hydrolysis. DAPA was accepted as a reliable marker for determining microbial protein in ileal digesta.

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Bacterial D-alanine concentrations as a marker of bacterial nitrogen in the gastrointestinal tract of pigs and cows

Cited by 8 publications

References 34 publications

2,6-Diaminopimelic acid as a biological marker of rumen synthesis and fermentation capacities in the transition period and early lactation of dairy cows

2,6-Diaminopimelic acid as a biological marker of rumen synthesis and fermentation capacities in the transition period and early lactation of dairy cows

Impact of diet composition on ileal digestibility and small intestinal morphology in early-weaned pigs fitted with a T-cannula

Comparison of three markers for the determination of bacterial protein in terminal ileal digesta in the growing pig

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