Nitrogen fractionation in faeces: status quo and potential
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1 Nitrogen fractionation in faeces: status quo and potential Marcus Clauss & Jürgen Hummel Clinic for Zoo Animals, Exotic Pets and Wildlife, Vetsuisse Faculty, University of Zurich, Switzerland Department of Animal Sciences, Ruminant Nutrition, Georg-August-University of Göttingen, Germany 75. Geburtstag Prof. Dr. Dr. h.c. Ernst Pfeffer, Bonn 2014
2 Apparent digestibility
3 Digestibility (Intake Excretion) / Intake
4 True digestibility (Intake Excretion) / Intake undigested diet remains
5 Apparent digestibility (Intake Excretion) / Intake undigested diet remains already metabolized ( metabolic ) substance
6 Apparent digestibility (Intake Excretion) / Intake undigested diet remains already metabolized ( metabolic ) substance endogenous substance
7 Apparent digestibility (Intake Excretion) / Intake undigested diet remains already metabolized ( metabolic ) substance endogenous substance microbial substance
8 Apparent digestibility (Intake Excretion) / Intake undigested diet remains already metabolized ( metabolic ) substance endogenous substance microbial substance If there is endogenous/ metabolic faecal excretion for a nutrient, its apparent digestibility increases with dietary concentration.
9 Apparent digestibility If there is endogenous/ metabolic faecal excretion for a nutrient, its apparent digestibility increases with dietary concentration.
10 Apparent digestibility ad Crude protein (%) Crude protein (% DM) If there is endogenous/ metabolic faecal excretion for a nutrient, its apparent digestibility increases with dietary concentration.
11 Apparent digestibility ad Crude protein (%) Crude protein (% DM) dig. Crude protein (% DM) Crude protein (% DM)
12 Estimating true digestibility and endogenous losses by regression analysis
13 Estimating true digestibility and endogenous losses by regression analysis
14 Estimating true digestibility and endogenous losses by regression analysis endogenous losses per 100 g DMI
15 Estimating true digestibility and endogenous losses by regression analysis true digestibility endogenous losses per 100 g DMI
16 Estimating true digestibility and endogenous losses by regression analysis
17 Estimating true digestibility and endogenous losses by regression analysis true digestibility endogenous losses per unit (metabolic) body mass for a specific diet
18 Estimating true digestibility and endogenous losses by regression analysis true digestibility for a specific diet endogenous losses per unit (metabolic) body mass for a specific diet
19 Estimating true digestibility and endogenous losses by regression analysis
20 Estimating true digestibility and endogenous losses by regression analysis Is there a conceptual consensus how regression analysis is used to estimate td and endogenous losses or is its use mainly determined by data availability?
21 Faecal nitrogen I - simple ecological applications (Total faecal nitrogen = TFN)
22 Faecal nitrogen Diet N Total faecal N undigested dietary N endogenous N MFN microbial N
23 TFN as a diet quality! indicator Diet N Total faecal N undigested dietary N endogenous N MFN microbial N
24 TFN as a diet quality! indicator Diet N Total faecal N!! undigested dietary N endogenous N MFN microbial N
25 TFN as a diet quality " indicator Diet N Total faecal N " " undigested dietary N endogenous N MFN microbial N
26 TFN as a diet quality " indicator Diet N Total faecal N " " undigested dietary N endogenous N MFN microbial N Because of a statistical correlation, TFN is most often interpreted as an indicator of diet N.
27 TFN as a diet quality! indicator
28 TFN in free-ranging populations
29 TFN in seasonal datasets
30 TFN in free-ranging ungulates from Steuer et al. (revision submitted)
31 TFN in zoo animals
32 Faecal nitrogen II - more elaborate ecological applications (Total faecal nitrogen = TFN)
33 TFN as a diet quality! indicator Diet N Total faecal N undigested dietary N endogenous N MFN microbial N
34 TFN is a concentration! Diet N Total faecal N undigested dietary N endogenous N MFN microbial N
35 TFN is a concentration! Diet N Total faecal N undigested dietary N endogenous N MFN microbial N other indigestible material
36 TFN as a diet quality! indicator Diet N Total faecal N undigested dietary N endogenous N MFN microbial N other indigestible material
37 TFN as a diet quality! indicator Diet N Easily fermentable carbohydrates Slowly fermenting fibre Indigestible fibre Total faecal N undigested dietary N endogenous N MFN microbial N other indigestible material
38 TFN as a diet quality! indicator Diet N Easily fermentable carbohydrates Slowly fermenting fibre Indigestible fibre Total faecal N undigested dietary N endogenous N MFN microbial N other indigestible material
39 TFN as a diet quality! indicator Diet N! Easily fermentable carbohydrates Slowly fermenting fibre Indigestible fibre Total faecal N undigested dietary N endogenous N MFN microbial N other indigestible material
40 TFN as a diet quality! indicator Diet N! Easily fermentable carbohydrates! Slowly fermenting fibre Indigestible fibre Total faecal N undigested dietary N endogenous N MFN microbial N other indigestible material
41 TFN as a diet quality! indicator Diet N! Easily fermentable carbohydrates! Slowly fermenting fibre Indigestible fibre " " Total faecal N undigested dietary N endogenous N MFN microbial N other indigestible material
42 TFN as a diet quality! indicator Diet N! Easily fermentable carbohydrates! Slowly fermenting fibre Indigestible fibre " " Total faecal N undigested dietary N endogenous N MFN microbial N " other indigestible material
43 TFN as a diet quality! indicator Diet N! Easily fermentable carbohydrates! Slowly fermenting fibre Indigestible fibre " " " Total faecal N undigested dietary N endogenous N MFN microbial N " other indigestible material
44 TFN as a diet quality! indicator Diet N! Easily fermentable carbohydrates! Slowly fermenting fibre Indigestible fibre " " " Total faecal N undigested dietary N endogenous N MFN microbial N " other indigestible material
45 TFN as a diet quality! indicator Diet N! Easily fermentable carbohydrates! Slowly fermenting fibre Indigestible fibre " " " Total faecal N undigested dietary N endogenous N MFN microbial N! " other indigestible material
46 TFN as a diet quality! indicator Diet N! Easily fermentable carbohydrates! Slowly fermenting fibre Indigestible fibre " " " Total faecal N undigested dietary N endogenous N MFN! microbial N! " other indigestible material
47 TFN as a diet quality! indicator Diet N! Easily fermentable carbohydrates! Slowly fermenting fibre Indigestible fibre " " " Total faecal N undigested dietary N endogenous N MFN!! microbial N! " other indigestible material
48 TFN as a diet quality! indicator Diet N! Easily fermentable carbohydrates! Slowly fermenting fibre Indigestible fibre " " " Total faecal N undigested dietary N endogenous N MFN!! microbial N! " other indigestible material TFN summarizes processes that are related to the overall digestion of the diet.
49 TFN as indicator of digestibility
50 TFN as indicator of digestibility
51 TFN as indicator of digestibility
52 TFN as indicator of digestibility
53 TFN as indicator of digestibility
54 Faecal nitrogen III - a fundamental constraint (Total faecal nitrogen = TFN)
55 TFN as indicator of digestibility
56 TFN and secondary plant compounds
57 TFN in comparative datasets
58 TFN in comparative datasets
59 TFN and secondary plant compounds
60 TFN and secondary plant compounds
61 Faecal nitrogen IV - fractionation (Total faecal nitrogen = TFN) (Metabolic faecal nitrogen = MFN)
62 Analytical approaches Total faecal N
63 Analytical approaches Total faecal N undigested dietary N MFN
64 Analytical approaches Total faecal N undigested dietary N MFN
65 Analytical approaches Total faecal N undigested dietary N MFN regression analysis
66 Analytical approaches Total faecal N undigested dietary N MFN regression analysis fibre-bound N
67 Analytical approaches Total faecal N undigested dietary N MFN regression analysis fibre-bound N NDF-N ADF-N Ultrasonic probe (ind.) Lysozyme-Trypsin extract (ind.)
68 Analytical approaches Total faecal N undigested dietary N MFN regression analysis fibre-bound N NDF-N ( undigested ) ADF-N ( indigestible ) Ultrasonic probe (ind.) Lysozyme-Trypsin extract (ind.)
69 Analytical approaches Total faecal N undigested dietary N MFN regression analysis endogenous N microbial N fibre-bound N NDF-N ADF-N Ultrasonic probe (ind.) Lysozyme-Trypsin extract (ind.)
70 Analytical approaches Total faecal N undigested dietary N MFN regression analysis endogenous N microbial N fibre-bound N NDF-N ADF-N Ultrasonic probe (ind.) Lysozyme-Trypsin extract (ind.) microbial N marker
71 Analytical approaches Total faecal N undigested dietary N MFN regression analysis endogenous N microbial N fibre-bound N NDF-N ADF-N Ultrasonic probe (ind.) Lysozyme-Trypsin extract (ind.) microbial N marker Diaminopimelic acid (DAPA) RNA extraction Purines Centrifugation Microscopy
72 MFN method Mason
73 MFN method Mason
74 MFN method Mason There is little evidence to link MFN to dietary consituents in larger-scale datasets (because of lack of data, not because of absence of link), although the logic appears sound.
75 Does MFN work and does it provide more information than TFN? (Total faecal nitrogen = TFN) (Metabolic faecal nitrogen = MFN)
76 In situ in the rumen: good results - Purine
77 In situ in the rumen: good results
78 TFN - DAPA
79 TFN/DAPA energy digestibility
80 TFN, MFN and other analytes in reaction to starch infusion
81 TFN and MFN at different diet quality and intake levels
82 MFN and faecal ADL
83 MFN in relation to faecal VFA
84 MFN in relation to dietary N
85 TFN/MFN fibre digestibility Comparing FN and the in vitro gas production of faecal fibre (inverse of digestibility achieved by the animal). from Steuer et al. (in prep.)
86 TFN/MFN fibre digestibility Comparing FN and the in vitro gas production of faecal fibre (inverse of digestibility achieved by the animal). from Steuer et al. (in prep.)
87 Foregut vs. Hindgut Fermentation from Stevens & Hume (1995)
88 Foregut vs. Hindgut Fermentation Lower bacterial nitrogen losses in the faeces? from Stevens & Hume (1995)
89 Foregut vs. Hindgut Fermentation Lower bacterial nitrogen losses in the faeces? Higher bacterial nitrogen losses in the faeces? from Stevens & Hume (1995)
90 Foregut vs. Hindgut Fermentation Lower bacterial nitrogen losses in the faeces? Lower bacterial nitrogen losses in hard faeces in coprophagic hindgut fermenters due to bacterial accumulation in caecotrophs? from Stevens & Hume (1995)
91 Foregut vs. Hindgut Fermentation Lower bacterial nitrogen losses in the faeces? Higher bacterial nitrogen losses in the faeces? Concentration in faeces vs. total excretion from Stevens & Hume (1995)
92 TFN in zoo animals
93 TFN in zoo animals
94 TFN in zoo animals from Hesta et al. (2003)
95 MFN in zoo animals
96 MFN in zoo animals
97 MFN in zoo animals from Hesta et al. (2003)
98 TFN MFN relationship: no influence of digestion type... except the most basic
99 TFN MFN relationship: no influence of digestion type... except the most basic
100 TFN MFN relationship: no influence of digestion type... except the most basic from Hesta et al. (2003)
101 TFN MFN relationship: no influence of digestion type from Steuer et al. (revision submitted)
102 TFN MFN relationship: no influence of digestion type from Steuer et al. (revision submitted)
103 from Steuer et al. (revision submitted) TFN/MFN on grass hay
104 from Steuer et al. (revision submitted) TFN/MFN on grass hay
105 MFN not better as indicator of digestibility
106 MFN not better as indicator of digestibility MFN
107 Does MFN work and does it provide more information than TFN in the presence of secondary compound? (Total faecal nitrogen = TFN) (Metabolic faecal nitrogen = MFN)
108 Faecal N and secondary compounds
109 TFN is influenced by diet GLM F p Body mass Digestion type (Rum vs. HF) from Steuer et al. (revision submitted)
110 TFN is influenced by diet GLM F p Body mass Digestion type (Rum vs. HF) %grass from Steuer et al. (revision submitted)
111 MFN is less influenced by diet GLM F p Body mass Digestion type (Rum vs. HF) %grass from Steuer et al. (revision submitted)
112 A failed attempt to quantify endogenous faecal N in herbivores
113 Analytical approaches Total faecal N undigested dietary N MFN regression analysis endogenous N microbial N fibre-bound N NDF-N ADF-N Ultrasonic probe (ind.) Lysozyme-Trypsin extract (ind.) microbial N marker Diaminopimelic acid (DAPA) RNA extraction Purines Centrifugation Microscopy
114 Analytical approaches Total faecal N undigested dietary N MFN regression analysis endogenous N microbial N fibre-bound N NDF-N ADF-N Ultrasonic probe (ind.) Lysozyme-Trypsin extract (ind.) vertebrate N marker microbial N marker Diaminopimelic acid (DAPA) RNA extraction Purines Centrifugation Microscopy
115 Analytical approaches Total faecal N undigested dietary N MFN regression analysis endogenous N microbial N fibre-bound N vertebrate N marker NDF-N (SDS difference?) ADF-N Ultrasonic probe (ind.) Lysozyme-Trypsin extract (ind.) microbial N marker Diaminopimelic acid (DAPA) RNA extraction Purines Centrifugation Microscopy
116 Analytical approaches Total faecal N undigested dietary N MFN regression analysis endogenous N microbial N fibre-bound N vertebrate N marker microbial N marker NDF (SDS) -N NDF (no SDS) -N
117 Used in a dog study
118 Pilot result...
119 ... but generally unplausible results
120 Applying MFN in an ecological context (Metabolic faecal nitrogen = MFN)
121 An old question: Do larger herbivores ingest lower-quality diets, and are they physiologically equipped for a better digestion of such diets?
122 Hypothesis building from Steuer et al. (revision submitted)
123 Larger size endows higher digestive efficiency... from Steuer et al. (revision submitted)
124 ... or body size has no effect on digestibility from Steuer et al. (revision submitted)
125 Body size does not affect diet selection... from Steuer et al. (revision submitted)
126 ... or larger animals eat lower quality diets from Steuer et al. (revision submitted)
127 ... or larger animals eat lower quality diets MFN (%OM) MFN (%OM) MFN (%OM) MFN (%OM) from Steuer et al. (revision submitted)
128 ... or larger animals eat lower quality diets MFN (%OM) MFN (%OM) MFN (%OM) MFN (%OM) from Steuer et al. (revision submitted)
129 ... or larger animals eat lower quality diets MFN (%OM) MFN (%OM) MFN (%OM) MFN (%OM) from Steuer et al. (revision submitted)
130 Conclusions
131 Conclusions!Total faecal nitrogen (TFN) is often used in ecology, but sometimes not with a functional understanding.
132 Conclusions!Total faecal nitrogen (TFN) is often used in ecology, but sometimes not with a functional understanding.!tfn reflects diet digestibility, but this is affected by plant secondary compounds (PSC).
133 Conclusions!Total faecal nitrogen (TFN) is often used in ecology, but sometimes not with a functional understanding.!tfn reflects diet digestibility, but this is affected by plant secondary compounds (PSC).!Quantifying metabolic faecal nitrogen (MFN) is possible using various approaches; NDF- or ADFsoluble N may be the most convenient.
134 Conclusions!Total faecal nitrogen (TFN) is often used in ecology, but sometimes not with a functional understanding.!tfn reflects diet digestibility, but this is affected by plant secondary compounds (PSC).!Quantifying metabolic faecal nitrogen (MFN) is possible using various approaches; NDF- or ADFsoluble N may be the most convenient.!simple assays to separate microbial and endogenous N have failed so far.
135 Conclusions!Total faecal nitrogen (TFN) is often used in ecology, but sometimes not with a functional understanding.!tfn reflects diet digestibility, but this is affected by plant secondary compounds (PSC).!Quantifying metabolic faecal nitrogen (MFN) is possible using various approaches; NDF- or ADFsoluble N may be the most convenient.!simple assays to separate microbial and endogenous N have failed so far.!herbivore digestive strategies are linked to digestibility and hence TFN or MFN, but not to differences in the MFN-TFN relationship.
136 Conclusions!Total faecal nitrogen (TFN) is often used in ecology, but sometimes not with a functional understanding.!tfn reflects diet digestibility, but this is affected by plant secondary compounds (PSC).!Quantifying metabolic faecal nitrogen (MFN) is possible using various approaches; NDF- or ADFsoluble N may be the most convenient.!simple assays to separate microbial and endogenous N have failed so far.!herbivore digestive strategies are linked to digestibility and hence TFN or MFN, but not to differences in the MFN-TFN relationship.!mfn appears superior to TFN in situations of heterogenous diets (incl. PSC).
137 thank you for your attention
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