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Indigestible components, including indigestible dry matter (iDM) and indigestible neutral detergent fiber (iNDF), play an integral role as internal markers for determining ruminal kinetics and digestibility estimations. However, the accuracy of internal markers is dependent upon the incubation technique utilized as bag type (BT) and incubation length (IL) can be significant sources of error. Previous studies have primarily focused on iDM and iNDF as digestibility markers, but few studies have compared digestibility estimates to those of acid detergent insoluble ash (ADIA). Therefore, our objective was to investigate the effect of BT (F57, F58, and Dacron) and IL (288 and 576 h) on iDM and iNDF residues, DM and NDF digestibilities, and fecal recoveries when using in situ incubations. Additionally, we evaluated the accuracy of digestibility estimates when using iDM, iNDF, and ADIA. For iDM and iNDF, feed residues demonstrated a BT × IL interaction (P < 0.01). However, fecal residues were only influenced by the main effects of BT and IL (P < 0.01), with the F58 BT and 288-h IL having the greatest residues for both iDM and iNDF. The variation in residues was greatly reduced when using iNDF compared with iDM. Fecal recovery estimates most closely approximated 100% recovery when utilizing ADIA and iDM using the F57 × 576 h incubation method (P < 0.01), although recovery was overestimated for all incubation combinations. Fecal NDF recovery estimates better represented the excretion profiles when the F57 × 576 h combination was used with iDM as the internal marker (P < 0.01). Estimates of DM and NDF digestibility were the most accurate when utilizing ADIA (P < 0.01) relative to all other treatments. Our results indicate that the proper methodological application is specific to the purpose of the inferences. When evaluating fecal recoveries and digestibility, ADIA or iDM with F57 at 576-h in situ incubation provides the greatest accuracy.

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RESUMEN

Reliable assessments of indigestible dietary components are required when using internal markers to estimate diet digestibility and determine the potentially digestible portion of the fiber. The lack of a standardized methodology and understanding of how antinutritional factors influence indigestible residues can result in erroneous estimates with inconsistent variation across trials and among studies. Previous studies have detailed suitable bag porosity and sample size (SS) with incubation length (IL) varying from 96 to 504 h, with many assuming that 288-h IL yields truly indigestible components. Recent studies have primarily investigated the variation that exists among feedstuffs, but most have failed to account for possible effects of secondary compounds. Using 2 similar concentrate diets, one of which contained supplemental condensed tannins (CT), we investigated the effect of bag type (BT; 10 and 25 µm), SS (20 and 40 mg/cm2), and IL (288 and 576 h) on in situ indigestible DM (iDM) and neutral detergent fiber (iNDF) residues of feed and feces, and resultant DM and NDF digestibilities. There were no 3-way interactions (P > 0.05), but 2-way interactions were present for iDM and iNDF residues with BT × SS influencing the control (no CT) ration (P < 0.01), SS × IL impacting feed containing CT (P < 0.01), and BT × IL affecting both feedstuffs (P ≤ 0.01). For the control diet, only BT × SS affected DM and NDF digestibilities. Whereas the CT diet did not demonstrate any significant interactions for digestibilities. Values of iDM were largely influenced by contamination that varied greatly based on intrinsic factors associated with the bag and incubation duration. The presence of CT influenced iDM and iNDF to varying degrees due to possible trapping of CT-substrate complexes. For the control diet, the use of 25-µm bags resulted in lower fecal recoveries relative to the 10 µm (P < 0.01). However, there appears to be a dynamic relationship among BT, SS, and IL within respective diets and sample types that can affect indigestible components and resultant digestibility estimates. Based on simulations from these data, the sample size required to attain 90% power when utilizing 2 incubation animals exceeds the triplicate and quadruplicate replications commonly utilized. Further emphasizing the necessity for a more complete understanding of incubation dynamics to design biologically and statistically valid investigations.

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Two experiments were conducted to evaluate the number of days required for total fecal collection and the viability of using the indigestible dry matter (iDM), indigestible neutral detergent fiber (iNDF), and indigestible acid detergent fiber (iADF) internal markers to determine the fecal excretion of dry matter (FEDM) and digestibility in nutritional trials with small ruminants. Eight sheep in the first experiment and eight goats in the second experiment were distributed into two 4 × 4 Latin square designs. There were no significant differences between days of total fecal collection for FEDM; digestibility of dry matter (DM), organic matter (OM), crude protein (CP), ether extract (EE), neutral detergent fiber corrected for ash and protein (NDFap), and non-fibrous carbohydrates corrected for ash and protein (NFCap); and total digestible nutrients (TDN) in both species. The results suggest that only 1 day of total collection is sufficient to obtain the FEDM and the digestibility of the nutritional components in sheep and goats. The markers are efficient in determining fecal production and digestibility in these animal species.

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BACKGROUND: The effect of time of fecal sampling on the accuracy of acid-detergent insoluble ash (ADIA) and alkaline-peroxide lignin (APL) for the prediction of fecal output (FO) in cattle was evaluated. Eight ruminally cannulated cows (594 ± 35.5 kg) were allocated randomly to 4 bermudagrass [Cynodon dactylon (L.) Pers.] hay diets markedly different in crude protein concentration (79-164 g/kg) with 2 replicates per diet for 3 periods. Cows were offered hay individually at 20 g DM/kg of body weight daily in equal feedings at 08:00 and 16:00 h for a 10-d adaptation period followed by 5-d of total fecal collection. Fecal grab samples also were taken each day during the fecal collection period at 06:00, 12:00, 18:00, and 24:00 h either directly from the rectum or from freshly voided feces. Samples were composited within cow and time across the 5 d total fecal collection period. Additionally, forage, ort, and fecal samples were analyzed for concentrations of APL and ADIA. RESULTS: Fecal concentrations of ADIA and APL were not affected by sampling time (P ≥ 0.22), even though diet affected (P < 0.01) fecal ADIA and APL concentrations. There were no diet × sampling time interactions (P ≥ 0.60). Estimates of FO and dry matter digestibility (DMD) from ADIA and APL were not affected (P ≥ 0.16) by sampling time or the diet × sampling time interaction (P ≥ 0.74). Estimates of FO and DMD from markers from different sampling times or all different combinations of sampling time were not different (P ≥ 0.72) from those of total collection among internal markers. CONCLUSION: Little variation in concentrations of ADIA and APL in daily fecal excretion across time increases flexibility in fecal grab sampling schedules for predicting FO and DMD.

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Quantificar a síntese de proteína microbiana em animais ruminantes é de grande importância para avaliar a qualidade nutritiva das dietas. A determinação dessa síntese se obtém por meio da utilização de marcadores microbianos, osquais são classificados em internos (subprodutos dos microrganismos ruminais) e externos (produtos administrados aos animais que são incorporados pelos microrganismos) e técnicas indiretas. Embora tenham ocorrido avanços ainda não se tem o marcador ideal. O objetivo desta revisão é detalhar a cada uma das vantagens e desvantagens das técnicas para estimar a síntese de proteína microbiana no rúmen e discutir os resultados obtidos utilizando cadatécnica.

The quantification of the microbial protein synthesis in the ruminants is of great importance to assess the nutritional quality of the diets. The determination of microbial synthesis is achieved through the use of microbial markers, which are classified: internal (by-products of rumen microorganisms) and external (substances administeredto animals that are incorporated by rumen microorganisms). Although some advances has occurred lately anideal marker still need to be developed. So the objective of this review is to detail each of the techniques forestimating microbial protein synthesis in the rumen, and discuss the results obtained using each technique.

La cuantificación de la síntesis de proteína microbiana en los rumiantes es de gran importancia para evaluar lacalidad nutricional de las dietas. La determinación de la síntesis microbiana se logra a través del uso de marcadoresmicrobianos que se clasifican como: Internos (subproductos de los organismos de la panza) y externos (sustancias administradas a los animales que son incorporadas por los organismos de la panza). Aunque últimamente ha habido algunos avances, aún es necesario desarrollar un marcador ideal. Así que el objetivo de esta revisión es detallarcada una de las técnicas usadas para estimar la síntesis de proteína microbiana en la panza y discutir los resultados obtenidos al usar cada técnica.

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