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The aim of this study was to check the accuracy of laboratory methods to predict the apparent protein digestibility (CPd, %) and digestible protein content (DP, g/kgDM) of dog foods, avoiding the use of experimental animals in digestion trials. Twenty-eight commercial dry extruded dog foods were tested by three different methodologies: an adaptation of the in vitro incubation method described by Hervera et al. (J Anim Physiol Anim Nutr 2007, 91: 205) for estimation of digestible energy of commercial dog foods, a modification of the pH drop methodology proposed by Hsu et al. (J Food Sci 1977, 42: 1269) for protein evaluation of human foods and the Near Infra Red Spectroscopy (NIRS) technology. All the methods assessed revealed very good, similar accuracy in the prediction of DP content either using the in vitro method (r = 0.99; RSD = 6.76; CV = 2.31%), the pH drop method (r = 0.99; RSD = 6.94; CV = 3.02%) or the NIRS (R2 = 0.96; SECV = 10.50) method, although the in vitro digestion method showed the highest accuracy approach of in vivo crude protein apparent digestibility: CPd in vitro (r = 0.81; RSD = 2.01; CV = 2.41%); CPd pH-drop (r = 0.78; RSD = 2.48; CV = 2.98%) and NIRS (r(2)cv = 0.53; SECV = 2.37).

The proposal of National Research Council (NRC), based on the use of modified Atwater factors, is nowadays the widely used method to estimate digestible energy (DE) content of pet foods. Recently, alternative methods have been suggested for predicting energy content of commercial canine dry food. Factorial equations including food fibre content as estimator, in vitro digestions methods or near-infrared spectroscopy (NIRS) techniques have been considered as good approaches to predict the energy content of dog foods. The aim of this study was to compare the accuracy of some of those estimation methods. Seventeen samples of commercial extruded dog food were used to validate and compare some estimation methods of energy digestibility (Ed, %) and DE value [MJ/kg dry matter (DM)]. The apparent Ed and DE of each food were previously determined by in vivo trials. In vivo Ed and DE of foods ranged from 79.30% to 91.05% and from 16.25 to 21.82 MJ/kg DM, respectively, and their crude fibre (CF) content ranged from 0.72% to 3.28% (in DM base). The % Ed of each sample was estimated by the factorial equation (% Ed = 91.2 - 1.43 x CF %) and by the in vitro digestion method [% Ed(in vitro) = -2.45 + 0.98 organic matter (OM) disappearance(in vitro)%]. The set of samples also was analysed by NIRS, using a calibration equation developed from a set of 69 samples of commercial extruded dog food (0.76 and 0.89 cross-validation r(2) and 2.33 and 0.61 cross-validation SE for Ed and DE respectively). The in vitro method gave better estimations of Ed in vivo than NIRS and factorial methods, although all the methods assessed showed a very good and similar accuracy in the prediction of DE value. These three methods showed a slight better accuracy than that previously proposed by the NRC. To consider constant digestibility values of nutrient content of food can result in bias and error in the estimated energy values. The alternative prediction methods used in this study take into account differences of ingredient composition and availability of nutrients of different extruded dog foods thus could be better systems of valuating energy content in a wider range of different kind of foods than in use method.

The aim of this study was to develop a simple and reproducible in vitro method for predicting the apparent energy digestibility of dry extruded dog foods. The proposed method is based on the two-step multienzymatic incubation assay described by [Boisen, S., 1991: In Vitro Digestion for Pigs and Poultry, M. F. Fuller (ed.). CAB International, Wallingford, 135-146], with some modifications adapted to dogs' digestion characteristics. The method consisted in two consecutives incubations, first one during 2 h with pepsin (10 mg/g of food sample) in acid pH and second one during 4 h with pancreatin (100 mg/g of food sample). The undigested residue obtained was collected in a filtration unit and then dried and ashed. The in vitro percentage of organic matter disappearance (in vitro dOM) of 54 dry extruded commercial dog foods was determined and used as predictor of the in vivo apparent organic matter (in vivo dOM) and energy digestibility (in vivo dE) and digestible energy (DE) content. There was a close linear relationship between the in vivo dOM and dE [r(2) = 0.95, residual standard deviation (RSD) = 1.05 and coefficient of variation (CV) = 1.2%] and also between the in vitro and in vivo dOM (r(2) = 0.92, RSD = 1.38 and CV = 1.6%), even if the in vitro dOM overestimated on average by 4% the in vivo dOM. When the in vitro dOM was used to predict the in vivo dE, the relationship between both variables was defined by the equation: in vivo dE (%) = -2.45 + 0.98 +/- 0.04x in vitro dOM (%), (r(2) = 0.92, RSD = 1.25 and CV = 1.5%). In addition, a close relationship between the in vivo and predicted DE (estimated dE x gross energy determined calorimetrically) was found (r(2) = 0.97, RSD = 0.26 and CV = 1.4%). The accuracy of DE content prediction using the proposed in vitro method was higher than that obtained when the DE content of the same set of samples was predicted by the equation proposed by the NRC (1985) (r(2) = 0.76, CV = 4.5%) and also slightly higher than that obtained when using the equation proposed by NRC (2006) (r(2) = 0.95, CV = 2.1%).

An experiment was performed to evaluate the effect of extrusion and carbohydrate addition on rumen degradation and intestinal digestion of raw legume seeds and solvent extracted soya bean meal (SBM) protein. Whole soya beans (WSB) without or with maize added (75:25) (WSB-M), peas, lupins and SBM were extruded at 140 degrees C. Protein rumen degradation and intestinal digestibility of unprocessed and extruded protein sources were measured by in sacco and mobile bag procedures, respectively, in two dairy cows cannulated in rumen and duodenum. Between 12 and 15 polyester bags with 4 g of each protein source were incubated in rumen for 12 h and the residues, pooled by feed, were introduced into the duodenum in small nylon bags after pre-incubation in a pepsin solution, and recovered from faeces the day after. Extrusion significantly (p < 0.001) reduced N degradation of all protein sources, from 98.1%, 91.6%, 90.5% and 64.8% to 53.1%, 73.8%, 70.3% and 44.2% for peas, lupins, WSB and SBM respectively. The addition of maize to WSB strengthened the effect of extrusion on rumen N degradation, from 88.2% to 52.6%. Residues from rumen incubation of extruded feeds showed a higher (p < 0.001) intestinal N digestibility except for SBM (87.0%, 82.9%, 66.3%, 85.0% and 97.2%, and 99.1%, 95.8%, 96.8%, 97.8% and 98.7%, respectively, for non-extruded and extruded, peas, lupins, WSB, WSB-M and SBM). In conclusion, the extrusion of studied legume seeds and SBM promotes a clear and significant increase of their metabolizable protein value, particularly in peas, and the inclusion of a source of carbohydrates before extrusion increase this response.

The effect of combinations of organic acids on digestive microbial population and total tract digestibility of piglets was studied. Thirty 19-21 days weaned pigs (5.4 +/- 0.23 kg, six pigs per treatment) were given a mixed feed with 10 ml water/kg (CTL) or 200 mEq/kg of formic acid (FOR), 1:1 formic:fumaric (FOFU), 1:1 formic:lactic (FOLA) or 2:1 formic:lactic (2FOLA). After 6-8 days, animals were slaughtered. In gastric contents, pH was higher (p = 0.01) with FOFU and lactobacilli tended (p = 0.08) to be lower with FOFU than CTL and FOLA, but coliform counts did not differ (p = 0.14). Acetate proportion was lower (p = 0.03), and propionate (p = 0.05) and butyrate (p = 0.01) higher, with FOFU than CTL, FOLA and 2FOLA. Intestinal coliform counts were higher (p = 0.03) with CTL than FOLA and 2FOLA, but there were no differences on lactobacilli. In intestinal contents, acetate tended (p = 0.06) to be lower with FOR than FOLA and 2FOLA, and butyrate was higher (p = 0.001) with FOR. Although not significantly, dry matter digestibility was 0.03-0.05 lower with CTL than with the other treatments. Combinations of organic acids in piglet diets modify gastric and intestinal microflora, the mixtures of formic:lactic appearing as the most interesting.

Near-infrared reflectance spectroscopy (NIRS) was used to predict the chemical composition, apparent digestibility and digestible nutrients and energy content of commercial extruded compound foods for dogs. Fifty-six foods of known chemical composition and in vivo apparent digestibility were analysed overall and 51 foods were used to predict gross energy digestibility and digestible energy content. Modified partial least square calibration models were developed for organic matter (OM), crude protein (CP), ether extract (EE), crude fibre (CF), nitrogen free extracts (NFE) and gross energy (GE) content, the apparent digestibility (OMD, CPD, EED, NFED and GED) and the digestible nutrient and energy content (DOM, DCP, DEE, DNFE and DE) of foods. The calibration equations obtained were evaluated by the standard error and the determination coefficient of cross-validation. The cross-validation coefficients of determination (R) were 0.61, 0.99, 0.91, 0.96, 0.94 and 0.92 for OM, CP, EE, CF, NFE and GE, the corresponding standard error of cross-validation (SECV) being 5.80, 3.51, 13.35, 3.64 and 16.95 g/kg dry matter (DM) and 0.29 MJ/kg DM respectively. The prediction of apparent digestibility was slightly less accurate, but NIRS prediction of digestible nutrient (g/kg DM) and DE (MJ/kg DM) gave satisfactory results, with high R (0.93, 0.97, 0.93, 0.83 and 0.93 for DOM, DCP, DEE, DNFE and DE respectively) and relatively low SECV (11.55, 6.85, 12.14 and 22.98 g/kg DM and 0.47 MJ/kg DM). It is concluded that the precision of NIRS in predicting the energy value of compound extruded foods for dogs is similar or better than by proximate analysis, as well as being faster and more accurate.

The apparent digestibility of nutrients and energy of 38 commercial dry extruded dog foods was measured using six adult (2 to 3 year-old) female Beagles. Diets contained [in g/kg dry matter (DM)]: 164-360 crude protein (CP); 79-261 ether extracts (EE); 8-33 crude fibre (CF) and 318-585 nitrogen free extracts (NFE). Apparent energy digestibility ranged from 77.3 to 91.6%, and was closely related to CF content (r=-0.85), yielding the resultant equation: GED (%)=94.00 - 4.04 x CF (% DM). The estimation of digestible energy content of foods from digestibility coefficients predicted from the above equation and gross energy measured or estimated from the Weende fractions, provides a more accurate prediction of experimental values than the Atwater approach followed by the National Research Council and the Association of American Feed Control Officials.

The present study examined the effect of urea-N supplementation of a N-deficient diet on digestion and metabolism in the rumen. Five Rasa Aragonesa ewes, each fitted with a rumen cannula, were offered alkali-treated barley straw ad lib. alone or supplemented continuously via the cannula with four levels of urea-N (3, 6, 9 and 12 g/d). Rumen NH3 concentrations increased in response to urea infusion (6-128 mg/l; P < 0.001). At the highest level of rumen NH3 concentration there was a significant increase, compared with the unsupplemented treatment, in dry matter (DM) intake (846-1206 g/d; P < 0.001) and apparent digestibility of DM (0.38-0.43), organic matter (0.38-0.45) and neutral-detergent fibre (0.41-0.49; P < 0.01). Rumen outflow rates of particulate matter and potential DM disappearances, assessed using nylon bags, were not affected by the experimental treatments, although fractional rate of DM disappearance increased significantly with increasing levels of urea infusion (2.4-4.6 per h). Urinary excretion of total purine derivatives increased with N supplementation, although the response was exclusively due to an increase in allantoin excretion (26.9-66.4 mg/kg live weight (W)0.75 per d; P < 0.001). Xanthine, hypoxanthine and uric acid excretion rates were constant, averaging 1.8 (SE 0.17); 5.4 (SE 0.21) and 7.2 (SE 0.36) mg/kg W0.75 per d respectively. The maintenance of a minimum rumen NH3 concentration (approximately 50 mg/l) was necessary to avoid significant reductions in DM intake and fermentation rate. Higher levels, however, may further increase microbial N flow at the duodenum, as suggested by the response in urinary allantoin excretion over the range of rumen NH3 concentrations.

Near-infrared spectroscopy (NIRS) is used in the pet food industry for rapid assessment of several macronutrients and GE content, but there is little published data on its usefulness for evaluating GE and GE digestibility (GED) of commercial pet food. Using NIRS spectra of 71 commercial extruded dog foods and reference values determined with calorimetry and in vivo feeding trials, chemometric models were developed for GE, GED, and DE prediction. The SE and R2 of cross-validation were 0.30 MJ/kg DM and 0.93 for GE, 2.10% and 0.82 for GED, and 0.53 MJ/kg DM and 0.92 for DE. The results indicated that NIRS provides GE, GED, and DE estimation values for dog food with an accuracy similar to that of the 2006 NRC proposed equations for use in pet food. Near-infrared spectroscopy is a fast and accurate method for predicting energy content in commercial extruded dog food, and is a useful and reliable tool to be used by the pet food industry when a wide enough calibration set is available. [ABSTRACT FROM AUTHOR]

An experiment was performed to evaluate the effect of extrusion and carbohydrate addition on rumen degradation and intestinal digestion of raw legume seeds and solvent extracted soya bean meal (SBM) protein. Whole soya beans (WSB) without or with maize added (75:25) (WSB-M), peas, lupins and SBM were extruded at 140 °C. Protein rumen degradation and intestinal digestibility of unprocessed and extruded protein sources were measured byin saccoand mobile bag procedures, respectively, in two dairy cows cannulated in rumen and duodenum. Between 12 and 15 polyester bags with 4 g of each protein source were incubated in rumen for 12 h and the residues, pooled by feed, were introduced into the duodenum in small nylon bags after pre-incubation in a pepsin solution, and recovered from faeces the day after. Extrusion significantly (p < 0.001) reduced N degradation of all protein sources, from 98.1%, 91.6%, 90.5% and 64.8% to 53.1%, 73.8%, 70.3% and 44.2% for peas, lupins, WSB and SBM respectively. The addition of maize to WSB strengthened the effect of extrusion on rumen N degradation, from 88.2% to 52.6%. Residues from rumen incubation of extruded feeds showed a higher (p < 0.001) intestinal N digestibility except for SBM (87.0%, 82.9%, 66.3%, 85.0% and 97.2%, and 99.1%, 95.8%, 96.8%, 97.8% and 98.7%, respectively, for non-extruded and extruded, peas, lupins, WSB, WSB-M and SBM). In conclusion, the extrusion of studied legume seeds and SBM promotes a clear and significant increase of their metabolizable protein value, particularly in peas, and the inclusion of a source of carbohydrates before extrusion increase this response. [ABSTRACT FROM AUTHOR]

Abstract: A feeding and digestion study was completed to determine whether sodium bicarbonate supplementation modifies diet selection of whole barley grain and a protein supplement by lambs. The underlying hypothesis was that aversion to barley intake to avoid acidosis might explain overconsumption of protein. Sixty weaned lambs (15.1±0.33kg live weight (LW)) were allocated to one of three dietary treatments in 2 pens of 5 lambs by sex, according to a 3 treatments×2 sex factorial design. Treatments were whole barley grain and a pelleted protein supplement fed at choice either unsupplemented (U) or supplemented with sodium bicarbonate fed by free access on the side (BF) or incorporated into the protein supplement at 15g/kg dry matter (BI). At the end of the growing period, 7 male lambs from each treatment (24.2±0.13kgLW) were moved to individual pens and slaughtered after 5 days of individual intake control to characterise rumen fermentation and determine digesta flow and microbial crude protein (CP) contribution to the abomasum, using C31 alkane and purine bases as flow and microbial markers, respectively. Intake of sodium bicarbonate was higher (P<0.005) when provided free access than when incorporated into the protein pellet (13.9g/d vs. 5.0g/d for BF and BI) but it did not affect dry matter intake (797g/d) or average daily gain (328g/d vs. 289g/d for male and female lambs), although barley grain intake tended (P=0.052) to decrease in response to buffer addition. A lower (P<0.05) proportion of organic matter truly digested in the rumen (OMTDR; 0.53 vs. 0.61) and a trend (P=0.098) to increased efficiency of microbial CP synthesis (26.7gN/kg vs. 21.6gN/kg OMTDR) ocurred when the buffer was offered free access compared to incorporation into the protein supplement, respectively. Neither rumen fluid pH nor ammonia concentration differed among treatments, but the total concentration of volatile fatty acids and molar proportion of acetate tended (P<0.1) to increase with buffer supplementation (174mM vs. 200mM and 46.8mol/100mol vs. 52.7mol/100mol). Sodium bicarbonate can modify rumen digestion, but failed to buffer rumen fluid pH and reverse the aversion to barley grain. [Copyright &y& Elsevier]