Effects of agroindustrial by-product supplementation on dairy goat milk characteristics, nutrient utilization, ruminal fermentation, and methane production.

The use of agroindustrial by-products, such as dried distillers grains with solubles (DDGS) and dried citrus pulp (DCP), has been widely investigated in dairy cows, but information on their effects in dairy goats is limited. The influence of feeding olive cake (a by-product of olive oil production) to dairy goats has been assessed in some studies, but exhausted olive cake (EOC) has been much less investigated. Twelve Murciano-Granadina goats were used in a crossover design trial with 2 periods to assess the effects of including agroindustrial by-products on nutrient digestibility, ruminal fermentation, methane production, urinary excretion of purine derivatives, and milk yield and composition. In each period, 6 goats received daily a control diet comprising 1 kg of alfalfa hay and 1 kg of high-cereal concentrate, and another 6 goats received a diet (BYP) comprising 1 kg of alfalfa hay and 1 kg of a concentrate including corn DDGS, DCP, and EOC in proportions of 180, 180, and 80 g/kg of concentrate (as-fed basis), respectively. Diet had no effect on total dry matter intake, but intake of alfalfa hay, CP, and fat was greater for the BYP group than for the control group. There were no differences between diets in nutrient apparent digestibility, with the exception of fat, which was greater for the BYP diet compared with the control diet. Although fecal N tended to be greater for the BYP diet, there were no differences in N utilization. Compared with the control diet, milk yield tended to be greater and daily production of milk CP, fat, whey protein, and TS as well as milk gross energy were greater for the BYP diet. The concentration of C12:0, C14:0, and C16:0 fatty acids (FA) was or tended to be lower and the concentration of polyunsaturated FA was greater in the milk of BYP-fed goats compared with goats fed the control diet. Diet had no effect on ruminal parameters (pH, volatile FA, and NH3-N concentrations) and methane emissions, but urinary excretion of total purine derivatives tended to be lower in BYP-fed goats than in those fed the control diet. A mixture of corn DDGS (180 g), DCP (180 g), and EOC (80 g) could replace 44% of cereal grains and protein feeds in the concentrate for dairy goats without compromising nutrient utilization, ruminal fermentation, or milk yield and led to a more unsaturated FA profile in milk.

Can by-products replace conventional ingredients in concentrate of dairy goat diet?

Intensive dairy goat production in the Mediterranean basin is based on imported conventional ingredients to be included in concentrates. Fourteen Murciano-Granadina goats in the middle of the third lactation were allocated into 2 groups of 7 animals each fed, respectively, a control diet based on alfalfa hay and concentrate in a 40:60 ratio, and a diet in which the concentrate included tomato fruits, citrus pulp, brewer's grain and brewer's yeast (T100CBY) to study the effect of diet on nutrient utilization, ruminal fermentation, purine derivatives excretion in urine, milk yield and composition, and methane emissions. No effect of the diet on total dry matter intake was observed. Digestibility of neutral detergent fiber and acid detergent fiber were higher for T100CBY compared with the control diet. The N in feces and urine was lower and balance and retained N were higher in animals fed T100CBY than the control diet. Milk protein N and energy were similar for both diets. Metabolizable energy per energy intake and metabolizable energy per digestible energy were higher and energy in methane was lower with diet T100CBY than with the control. Milk yield and composition were not affected by diet, with the exception of protein, casein, and total solids, which were higher for diet T100CBY than the control. Diet T100CBY promoted less saturated fatty acids and higher mono- and polyunsaturated fatty acids in milk than the control diet. Diet T100CBY produced less methane and NH3 concentration in the rumen, higher propionate, and a lower acetate-to-propionate ratio without an effect on the volatile fatty acid concentration. The concentrate with by-products did not affect urinary excretion of total purine derivatives, reduced feeding costs, and increased profit margin by 14 and 16% compared with the control. The mixture of tomato fruits, citrus pulp, brewer's grain, and brewer's yeast could replace 47% of conventional ingredients (corn, wheat bran, sunflower meal, and soy flour) in the concentrate of the dairy goat diet, reducing feeding cost and methane production, leading to a healthier fatty acids profile in milk without compromising nutrient utilization or milk yield.

Effects of ethyl-3-nitrooxy propionate and 3-nitrooxypropanol on ruminal fermentation, microbial abundance, and methane emissions in sheep.

The aim of this work was to investigate the effect of feeding ethyl-3-nitrooxy propionate (E3NP) and 3-nitrooxypropanol (3 NP), 2 recently developed compounds with potential antimethanogenic activity, in vitro and in vivo in nonlactating sheep on ruminal methane production, fermentation pattern, the abundance of major microbial groups, and feed degradability. Three experiments were conducted, 1 in vitro and 2 in vivo. The in vitro batch culture trial (experiment 1) tested 2 doses of E3NP and 3 NP (40 and 80 µL/L), which showed a substantial reduction of methane production (up to 95%) without affecting concentration of volatile fatty acids (VFA). The 2 in vivo trials were conducted over 16 d (experiment 2) and 30 d (experiment 3) to study their effects in sheep. In experiment 2, 6 adult nonpregnant sheep, with permanent rumen cannula and fed alfalfa hay and oats (60:40), were treated with E3NP at 2 doses (50 and 500 mg/animal per day). After 7, 14, and 15 d of treatment, methane emissions were recorded in respiration chambers and rumen fluid samples were collected for VFA analysis and quantification of bacterial, protozoal, and archaeal numbers by real-time PCR. Methane production decreased by 29% compared with the control with the higher dose of E3NP on d 14 to 15. A decrease in the acetate:propionate ratio was observed without detrimental effects on dry matter intake. In experiment 3, 9 adult nonpregnant sheep, with permanent rumen cannula and fed with alfalfa hay and oats (60:40), were treated with E3NP or 3 NP at one dose (100mg/animal per day) over 30 d. On d 14 and d 29 to 30, methane emissions were recorded in respiration chambers. Rumen fluid samples were collected on d 29 and 30 for VFA analysis and quantification of bacterial, protozoal, and archaeal numbers by real-time PCR. In addition, on d 22 and 23, samples of oats and alfalfa hay were incubated in the rumen of sheep to determine dry matter ruminal degradation over 24 and 48 h, respectively; no effect was observed (78.6, 78.3, and 78.8% of alfalfa and 74.2, 74.0, and 70.6% of oats in control, E3NP, and 3 NP groups, respectively). A reduction in methane production was observed for both additives at d 14 and d 29 to 30. In both treatments, the acetate:propionate ratio was significantly decreased. Likewise, total concentrations of the analyzed microbial groups in the rumen showed no difference among treatments and doses for both experiments. Both tested compounds showed promise as methane inhibitors in the rumen, with no detrimental effects on fermentation or intake, which would need to be confirmed in lactating animals.

Microbial and chemical composition of liquid-associated bacteria in goats' rumen and fermenters.

This study was undertaken to investigate the relationship between chemical composition and microbial profile of rumen liquid-associated bacteria (LAB) in vivo (Murciano-Granadina goats) and in a rumen simulation system (single-flow continuous-culture fermenters). To achieve this aim, analyses of purine bases along with some molecular techniques (quantitative PCR to assess abundance and DGGE to identify biodiversity and bacterial profile) were carried out. A control diet (AHC) based on alfalfa hay (AH) and concentrate (C) in a 1:1 ratio and two experimental diets (AHCBI and AHCBII), in which concentrate was partially replaced with multinutrient blocks, were used. Diets AHCBI and AHCBII included multinutrient blocks differing in the relative amount of two-stage olive cake and the source of protein (sunflower meal vs. fava beans). We aimed to investigate the effect of these blocks on rumen microbiota to evaluate their potential as safe substitutes of cereal-based concentrates. Similar patterns of response to diet were found for chemical composition, microbial abundances and diversity in LAB isolated from goat's rumen and fermenters. Whereas bacterial density (log10 gene copies/g FM: 11.6 and 9.4 for bacteria and methanogens, respectively, in rumen) and diversity indexes (Shannon index: 3.6) were not affected by diet, DGGE analyses showed that bacterial community profile was affected. The cluster analysis suggested differences in bacterial profile between LAB pellets isolated from the rumen of goat and fermenters. A relationship between chemical composition and bacterial community composition in LAB pellets seems to exist. Changes in the former were reflected in the bacterial community profile. Further research is needed to clarify the relationship between chemical and microbial composition of ruminal bacterial pellets with diets of different quality.

Dietary carbohydrate composition modifies the milk N efficiency in late lactation cows fed low crude protein diets.

Nitrogen emissions from dairy cows can be readily decreased by lowering the dietary CP concentration. The main objective of this work was to test whether the milk protein yield reduction associated with low N intakes could be partially compensated for by modifying the dietary carbohydrate composition (CHO). The effects of CHO on digestion, milk N efficiency (milk N/N intake; MNE) and animal performance were studied in four Jersey cows fed 100% or 80% of the recommended protein requirements using a 4×4 Latin square design. Four iso-energetic diets were formulated to two different CHO sources (starch diets with starch content of 34.3% and NDF at 32.5%, and fiber diets with starch content of 5.5% and NDF at 49.1%) and two CP levels (Low=12.0% and Normal=16.5%). The apparent digestible organic matter intake (DOMI) and the protein supply (protein digestible in the small intestine; PDIE) were similar between starch and fiber diets. As planned, microbial N flow (MNF) to the duodenum, estimated from the urinary purine derivatives (PD) excretion, was similar between Low and Normal CP diets. However, the MNF and the efficiency of microbial synthesis (g of microbial N/kg apparently DOMI) were higher for starch v. fiber diets. Milk and milk N fractions (CP, true protein, non-protein N (NPN)) yield were higher for starch compared with fiber diets and for Normal v. Low CP diets. Fecal N excretion was similar across dietary treatments. Despite a higher milk N ouput with starch v. fiber diets, the CHO modified neither the urinary N excretion nor the milk urea-N (MUN) concentration. The milk protein yield relative to both N and PDIE intakes was improved with starch compared with fiber diets. Concentrations of ß-hydroxybutyrate, urea and Glu increased and those of glucose and Ala decreased in plasma of cows fed starch v. fiber diets. On the other hand, plasma concentration of albumin, urea, insulin and His increased in cows fed Normal compared with Low CP diets. This study showed that decreasing the dietary CP proportion from 16.5% to 12.0% increases and decreases considerably the MNE and the urinary N excretion, respectively. Moreover, present results show that at similar digestible OM and PDIE intakes, diets rich in starch improves the MNE and could partially compensate for the negative effects of Low CP diets on milk protein yield.

In vitro-in vivo study on the effects of plant compounds on rumen fermentation, microbial abundances and methane emissions in goats.

Two in vitro and one in vivo experiments were conducted to investigate the effects of a selection of plant compounds on rumen fermentation, microbial concentration and methane emissions in goats. Treatments were: control (no additive), carvacrol (CAR), cinnamaldehyde (CIN), eugenol (EUG), propyl propane thiosulfinate (PTS), propyl propane thiosulfonate (PTSO), diallyl disulfide (DDS), a mixture (40 : 60) of PTS and PTSO (PTS+PTSO), and bromochloromethane (BCM) as positive control with proven antimethanogenic effectiveness. Four doses (40, 80, 160 and 320 µl/l) of the different compounds were incubated in vitro for 24 h in diluted rumen fluid from goats using two diets differing in starch and protein source within the concentrate (Experiment 1).The total gas production was linearly decreased (P<0.012) by all compounds, with the exception of EUG and PTS+PTSO (P≥ 0.366). Total volatile fatty-acid (VFA) concentration decreased (P≤ 0.018) only with PTS, PTSO and CAR, whereas the acetate:propionate ratio decreased (P≤ 0.002) with PTS, PTSO and BCM, and a tendency (P=0.064) was observed for DDS. On the basis of results from Experiment 1, two doses of PTS, CAR, CIN, BCM (160 and 320 µl/l), PTSO (40 and 160 µl/l) and DDS (80 and 320 µl/l) were further tested in vitro for 72 h (Experiment 2). The gas production kinetics were affected (P≤ 0.045) by all compounds, and digested NDF (DNDF) after 72 h of incubation was only linearly decreased (P≤ 0.004) by CAR and PTS. The addition of all compounds linearly decreased (P≤ 0.009) methane production, although the greatest reductions were observed for PTS (up to 96%), DDS (62%) and BCM (95%). No diet-dose interaction was observed. To further test the results obtained in vitro, two groups of 16 adult non-pregnant goats were used to study in vivo the effect of adding PTS (50, 100 and 200 mg/l rumen content per day) and BCM (50, 100 and 160 mg/l rumen content per day) during the 9 days on methane emissions (Experiment 3). The addition of PTS and BCM resulted in linear reductions (33% and 64%, respectively, P≤ 0.002) of methane production per unit of dry matter intake, which were lower than the maximum inhibition observed in vitro (87% and 96%, respectively). We conclude that applying the same doses in vivo as in vitro resulted in a proportional lower extent of methane decrease, and that PTS at 200 mg/l rumen content per day has the potential to reduce methane emissions in goats. Whether the reduction in methane emission observed in vivo persists over longer periods of treatments and improves feed conversion efficiency requires further research.

Nutrient utilization, ruminal fermentation, microbial nitrogen flow, microbial abundances, and methane emissions in goats fed diets including tomato and cucumber waste fruits.

The effects of replacing 50% of cereals-based concentrate with feed blocks (FB) including wastes of tomato fruits (diet ACT), wastes of cucumber fruits (diet ACC), or barley grain (diet ACB) on nutrient use, ruminal fermentation, microbial N flow to the duodenum, methane emissions, and abundances of total bacteria and methanogen were studied in goats. Four adult, dry, nonpregnant, rumen-fistulated Granadina goats (32.1 ± 5.52 kg BW) were used and 4 diets were studied in 4 trials by using a 4 × 4 Latin square experimental design. Diets consisted of alfalfa hay plus concentrate in a 1:1 ratio (AC) or 1:0.5 plus FB including wastes of ACT, ACC, or ACB. In each trial, goats were randomly assigned to 1 of the diets. Intake of each FB including ACT, ACC, and ACB were 203 ± 73, 179 ± 39, and 144 ± 68 g·animal(-1)·d(-1), respectively. The ACT and ACC diets did not (P > 0.05) compromise digestible and metabolizable energy, but reduced N retention by up to 29% (P = 0.03). Cucumber-based FB decreased the purine bases (PB):N ratio (P = 0.02) in total bacterial pellets isolated from the rumen content. Tomato-based FB decreased purine derivatives urinary excretion (P = 0.04) and microbial N flow (P = 0.03) without affecting the efficiency of synthesis but decreased methane emission by 28% (P < 0.001) compared with the other diets. Tomato and cucumber-based FB resulted in greater (P < 0.001) rumen VFA concentration and molar proportions of propionate and butyrate (P ≤ 0.02) than AC and ACB diets. No effect (P = 0.07) of diet on total bacteria abundance was observed whereas the abundance of methanogens increased (P = 0.01) with wastes-based FB. Our study suggests that ACC-based FB could replace one-half of the amount of concentrate in goat diet without compromising rumen fermentation and nutrient and energy use and without increasing methane emissions. Blocks including ACT showed an antimethanogenic effect but reduced microbial N flow to the duodenum. Further research is needed to improve both N and energy use of diets including ACT- and ACC-based FB.

Nutrient utilization, ruminal fermentation, microbial abundances, and milk yield and composition in dairy goats fed diets including tomato and cucumber waste fruits.

The effects of replacing 35% of cereals-based concentrate with feed blocks (FB) containing waste fruits of tomato, cucumber, or barley grain in diets for lactating goats on nutrient utilization, ruminal fermentation, microbial N flow to the duodenum, milk yield and quality, methane emissions, and abundances of total bacteria and methanogens were studied. Eight Murciano-Granadina goats (39.4 ± 5.39 kg of body weight, mean ± SD) in the middle of the third lactation were used and 4 diets were studied in a replicated 4×4 Latin square experimental design. Diets consisted of alfalfa hay (A) plus concentrate (C) in a 1:1 ratio (diet AC) or diets in which 35% of the concentrate was replaced with FB including wastes of tomato fruit, cucumber, or barley. In each period, 2 goats were randomly assigned to 1 of the dietary treatments. Intakes of FB including tomato, cucumber, and barley were 208 ± 65, 222 ± 52, and 209 ± 83 g of dry matter per animal and day, respectively. The replacement of 35% of concentrate with FB did not compromise nutrient apparent digestibility, total purine derivative urinary excretion, milk yield and composition, and total bacteria and methanogen abundances. Digestible energy and that in methane and urine were higher for AC than for FB-containing diets, whereas the metabolizable energy value was not affected by diet. The inclusion of tomato and cucumber fruits in FB decreased N in urine and CH(4) emissions compared with AC, which is environmentally relevant. However, tomato-based FB decreased microbial N flow in the rumen, whereas goats fed cucumber-based FB had the highest values for this measurement. Moreover, FB containing barley or tomato and cucumber led to lower rumen volatile fatty acid and NH(3)-N concentrations, respectively. Milk from goats fed diets including tomato and cucumber-based FB had higher linoleic, linolenic, and total polyunsaturated fatty acid concentrations than that from goats fed AC. Overall, our study suggests that tomato and cucumber FB could replace 35% of the concentrate in the dairy goat diet, reducing animal feeding cost and methane production, leading to higher polyunsaturated fatty acid proportions in milk, and without compromising nutrient utilization or milk yield.

Urinary excretion of purine derivatives, microbial protein synthesis, nitrogen use, and ruminal fermentation in sheep and goats fed diets of different quality.

The objective of this study was to compare N balance, microbial N flow (MNF) estimated from purine derivatives (PD) urinary excretion, and its variation when estimated using purine bases:N ratios in liquid associated bacteria (LAB) from models reported in the literature (MNF - response models) or measured ratios in liquid and solid-associated bacterial (SAB) pellets (MNF-LAB+SAB), diet digestibility, and rumen fermentation variables in sheep and goats fed 3 different practical, quality diets to study interspecies differences concerning N use as accurately as possible. Four mature female Merino sheep and 4 mature female Granadina goats, each fitted with a ruminal cannula, were used in 3 × 3 Latin square design with an extra animal. Two experimental diets had a forage-to-concentrate ratio of 70:30 (DM basis) with alfalfa hay (ALC) or grass hay (GRC) as forage, and the third diet contained 70% concentrate and 30% alfalfa hay (CAL). All animals were fed the diets at a daily rate of 56 g/kg BW(0.75) to minimize feed selection. Digestibility of nutrients was similar (P = 0.16 to 0.88) in the 2 species, but some animal species × diet interactions (P = 0.01 to 0.04) were detected. There were small differences between the fermentation patterns of both animal species. Goats showed decreased VFA concentrations (P = 0.005) and butyrate proportions (P = 0.04), and greater acetate proportions (P = 0.02) compared with sheep, whereas N intake and percentage of N intake excreted in feces were similar in both species (P = 0.58 and 0.15, respectively), the percentage excreted via the urine was greater in goats compared with sheep (P < 0.001). As a consequence, sheep had greater (P < 0.001) N retention than goats (averaged across diets, 32.6% and 16.1% of N intake, respectively). There were no differences (P = 0.95) between animal species in total PD excretion, but goats showed a greater excretion of allantoin (P = 0.01) and decreased excretion of xanthine (P = 0.008) and hypoxanthine (P = 0.007) compared with sheep. In general, differences between sheep and goats were more pronounced for the medium-quality diet (GRC) compared with those of high-quality diet (ALC and CAL). The greater urinary losses in goats would indicate a greater contribution of goats to N environmental contamination compared with sheep.

Effect of bromochloromethane on methane emission, rumen fermentation pattern, milk yield, and fatty acid profile in lactating dairy goats.

Several technologies have been tested to reduce enteric methanogenesis, but very few have been successfully used in practical conditions for livestock. Furthermore, the consequences of reduced rumen methane production on animal performance and milk quality are poorly understood. The aim of this work was to investigate the effect of feeding bromochloromethane (BCM), a halogenated aliphatic hydrocarbon with potential antimethanogenic activity, to dairy goats on rumen methane production, fermentation pattern, the abundance of major microbial groups, and on animal performance and milk composition. Eighteen goats were allocated to 2 experimental groups of 9 animals each: treated (BCM+) or not (BCM-) with 0.30 g of BCM/100 kg of body weight per day. The BCM was administered per os in 2 equal doses per day from parturition to 2 wk postweaning (10 wk). After weaning, methane emissions were recorded over 2 consecutive days (d 57 and 58 on treatment) in polycarbonate chambers. On d 59, individual rumen fluid samples were collected for volatile fatty acid (VFA) analysis and quantification of bacterial, protozoal, and archaeal numbers by real-time PCR. On d 69 and 70, daily milk production was recorded and samples were collected for determination of fat, protein, lactose, casein, and total solids concentration by infrared spectrophotometry, and fatty acid composition by gas chromatography. Treatment with BCM reduced methane production by 33% (21.6 vs. 14.4 L/kg of DMI) compared with nontreated animals, although it did not affect the abundance of rumen bacteria, protozoa, and total methanogenic archaea. The observed improvement in the efficiency of digestive processes was accompanied by a 36% increase in milk yield, probably due to the more propionic type of rumen fermentation and an increase in VFA production. The increase in milk yield was not accompanied by any changes in the concentrations or yields of fat, protein, or lactose. Despite the substantial decrease in methane production, only minor changes in milk fatty acid profile were observed, suggesting that ruminal biohydrogenation pathways were not affected. Compounds that influence terminal biochemical pathways for methane production deserve further development for future application in the dairy goat sector.

The effect of the feed-to-buffer ratio on bacterial diversity and ruminal fermentation in single-flow continuous-culture fermenters.

Eight single-flow continuous-culture fermenters were used in a completely randomized block design with a 2 × 4 factorial arrangement of treatments to investigate the effects of the feed-to-buffer ratio (F/B) on ruminal fermentation, the diversity and community structure of bacteria, nutrient digestibility, and N metabolism. Four diets with forage-to-concentrate ratios of 70:30 or 30:70 with alfalfa or grass hay as forage were supplied to fermenters twice per day at 2 different F/B (23.5 and 35 g of DM/L). The dilution rate was kept constant (5.3%) among all fermenters by infusing the same volume of buffer. An increase in the total volatile fatty acid (VFA) concentration and a decrease in the average pH were observed with an increased F/B. In addition, the molar proportions of all individual VFA found in fermenters differed, depending on the F/B. A terminal restriction fragment length polymorphism analysis showed that the community structure and diversity of bacteria were highly influenced by the F/B. Both diversity and the number of peaks in the electropherograms were lower in most fermenters receiving diets at a high F/B, whereas the similarity percentage of the bacterial communities across diets was higher as the F/B increased. Moreover, the high reduction of neutral detergent fiber digestibility (15.3% ± 3.65) in fermenters with high F/B suggested a pH-related decrease in the cellulolytic bacterial community as the F/B increased. The crude protein degradation found in fermenters receiving diets with a high F/B was lower compared with that from fermenters with a low F/B. The VFA concentration and purine bases flow response patterns to diets were similar to in vivo conditions only in the case of fermenters with a low F/B. The results suggested that the community structure and diversity of bacteria, as well as the in vitro fermentation parameters, may be affected by the F/B that is used, most likely through a pH effect. In addition, several fermentation parameters showed different response patterns to diets according to the F/B used. Therefore, the amount of feed supplied to single-flow continuous-culture fermenters in which pH is not under control should be carefully chosen according to the volume of buffer infused for the purpose of simulating ruminal fermentation.

Effects of partial replacement of concentrate with feed blocks on nutrient utilization, microbial N flow, and milk yield and composition in goats.

Two experiments were carried out to evaluate the effect of partial replacement of concentrate with 2 types of feed blocks (FB) on rumen protozoa numbers, nutrient utilization, microbial N flow to the duodenum, and milk yield and composition in goats. The concentrate included oat, corn grain, barley, soybean meal, salt, and vitamin-mineral mixture. The FB (types I and II) were composed of crude 2-stage olive cake (120 or 100g/kg), fava beans (0 or 400g/kg), barley (320 or 200g/kg), beet molasses (220 or 100g/kg), sunflower meal (180 or 0g/kg), quicklime (70 or 90g/kg), salt (60g/kg), urea (0 or 20g/kg), and vitamin-mineral mixture (30g/kg). In experiment 1, 6 adult, dry, nonpregnant, rumen-fistulated Granadina goats (46.9+/-2.15kg of BW) were used and 3 trials were carried out. In each trial, 2 goats were randomly assigned to receive 600g of alfalfa hay and 400g of concentrate (diet AC), 600g of alfalfa hay, 200g of concentrate, and FB I (diet ACBI), or 600g of alfalfa hay, 200g of concentrate, and FB II (diet ACBII) with 6 replications per diet. The FB were supplied ad libitum. The ratio of purine bases to N was higher in solid- and liquid-associated bacteria for FB goats than for AC goats. In experiment 2, 18 Granadina goats (39.6+/-1.89kg of BW) in the middle of the third lactation were used, and 3 trials were carried out by following a 3 x 3 Latin square experimental design. In every trial, 6 animals randomly received 1.0kg of alfalfa hay supplemented with 1.0kg of concentrate (diet AC) or 0.5kg of concentrate and FB I and II (diets ACBI and ACBII) with 18 replications per diet. The FB were supplied ad libitum. The intakes of organic matter and fat were higher with the AC diet than with the FB diets. The intake of acid detergent fiber was higher for FB-containing diets than for the AC diet. The neutral detergent fiber digestibility of FB diets was higher than that of the AC diet. Energy intake was higher for diets AC and ACBII than for ACBI. Microbial N flow was affected by diet. Milk yield was higher in goats fed the AC diet than in those receiving the FB diets. Conjugated linoleic acid content was higher in milk from FB than in milk from AC goats. Our study suggests that FB type II based on local ingredients could be used advantageously to reduce half of the amount of concentrate without detrimental effects on nutrient utilization, N value of the diet, and milk composition. The decrease of milk yield with ACBII compared with that obtained with the AC diet could be compensated by better quality of milk, decreased cost of feeding, and environmental advantage derived of including by-products in FB.

Nitrogen and energy utilization in lactating dairy goats fed diets with different legume seeds.

The N and energy utilization of diets with different legume seeds was analysed in lactating goats. Four isonitrogenous and isoenergetic diets were used and in each case 30% of the protein was supplied by one of four different legume seeds: lupins (L), faba beans (FB), bitter vetch (BV) and vetch (V). A group of eight Granadina goats, allocated to a replicated 4 × 4 Latin Square, were used. The N and energy utilization of the different diets may be considered adequate; milk N/intake N was approximately 0.20, milk N/N balance was approximately 0.50 and milk energy/metabolizable energy was approximately 0.30. Under similar N and energy intakes, faecal and urinary excretion of N and the quantities in milk did not vary significantly. However, the partition of the N balance between milk production and deposit within the body varied depending on the diet. Although this N available to the metabolism was the highest for the FB diet, the quantity of this directed to milk was the lowest. Finally, the faecal and urinary excretion with respect to the quantity directed to milk production did not vary significantly among the different diets.

Nutritive evaluation of legume seeds for ruminant feeding.

Chemical composition, rumen degradability and the effect of particle losses, and intestinal digestibility of protein by using in situ-in vitro and in vitro techniques were stated for beans (Vicia faba), lupin (Lupinus albus), vetch (Vicia sativa) and bitter vetch (Vicia ervilia) and four diets including those legume seeds. In addition, the apparent digestibility of experimental diets was determined in goats. The legume seeds showed high protein content (206-319 g/kg dry matter). Effective degradability of protein for legumes and diets varied from 0.80 to 0.87 and 0.76 to 0.82, respectively, decreasing to 0.53-0.76 and 0.61-0.67, respectively, when particle loss was taken into account. Different intestinal digestibility values were obtained with both methodologies without significant relationship between them (y = 1.058-0.463x; R(2)=0.068; RSD = 0.140; p = 0.53). There were no differences in the apparent nutrients and energy digestibility among diets (p > 0.05). These legumes can supply rapidly degradable protein for microbial protein synthesis and contribute to the pool of amino acids available for the synthesis of milk protein and for retention in the body.

Ruminal fermentation, microbial growth and amino acid flow in single-flow continuous culture fermenters fed a diet containing olive leaves.

Six single-flow continuous culture fermenters were used to determine fermentation profile, microbial growth and amino acid (AA) flow promoted by olive leaves supplemented with barley grains and faba beans (OLSUP), and alfalfa hay (AH). Two incubation runs were carried out with three fermenters inoculated with ruminal fluid from wethers and three from goats. The inoculum source did not affect (p = 0.059 to 0.980) any of the parameters. Daily volatile fatty acid (VFA) production and carbohydrate digestibility were greater (p = 0.009 and 0.024, respectively) for AH, therefore the pH values were lower (p = 0.015) than for OLSUP. Acetate was greater (p < 0.001) and isobutyrate, isovalerate and caproate lower (p < 0.001 to 0.006) for AH with greater acetate/propionate (p = 0.014) and 'VFA/digested carbohydrate' (p = 0.026) ratios. Daily microbial N flow and efficiency were greater (p = 0.016 and p = 0.041) for diet AH. Individual AA flows were greater (p < 0.001 to 0.016) for AH, but microbial essential AA proportion was greater for OLSUP (p = 0.015). The results indicate that OLSUP promoted lower bacterial growth and AA flow than AH, which could have been partially due to a limitation of N availability to ruminal microbes.

Effects of forage:concentrate ratio and forage type on apparent digestibility, ruminal fermentation, and microbial growth in goats.

The effects of forage type and forage:concentrate ratio (F:C) on apparent nutrient digestibility, ruminal fermentation, and microbial growth were investigated in goats. A comparison between liquid (LAB) and solid (SAB)-associated bacteria to estimate microbial N flow (MNF) from urinary purine derivative excretion was also examined. Treatments were a 2 x 2 factorial arrangement of forage type (grass hay vs. alfalfa hay) and high vs. low F:C (70:30 and 30:70, respectively). Four ruminally cannulated goats were fed, at maintenance intake, 4 experimental diets according to a 4 x 4 Latin square design. High-concentrate diets resulted in greater (P < 0.001) nutrient digestibility except for ADF. However, CP digestibility increased (P < 0.001) only for the high-concentrate diets including grass hay. Likewise, N retention, ruminal NH(3)-N concentration, and urinary excretion of purine derivatives increased (P < 0.05) with increasing concentrate in animals fed diets based on grass hay (0.23 vs. 0.13 g of retained N/g of digested N, 30.1 vs. 12.9 mg of NH(3)-N/100 mL, and 11.5 vs. 8.40 mmol/d, respectively), but not (P > 0.05) when diets included alfalfa hay. Total protozoa numbers and holotricha proportion were greater and less (P < 0.001), respectively, in high- than in low-concentrate diets. The F:C affected (P < 0.001) ruminal pH but not total VFA concentration (P = 0.12). Ammonia-N concentration was similar (P = 0.13) over time, whereas pH, VFA concentration, and protozoa numbers differed (P < 0.001) among diets. Estimated MNF was strongly influenced by using either the purine bases:N ratio obtained in our experimental conditions or values reported in the literature for small ruminants. There was a F:C effect (P = 0.006) on MNF estimated from LAB but not from SAB. The effect of F:C shifting from 70:30 to 30:70 in goat diets depends on the type of forage used. The MNF measured in goats fed different diets was influenced by the bacterial pellet (LAB or SAB). In addition, the purine bases:N ratio values found were different from those reported in the literature, which underlines the need for these variables to be analyzed directly in pellets isolated from specific animals and experimental conditions.

Comparison of bacterial pellets and microbial markers for the estimation of the microbial nitrogen and amino acids flows from single flow continuous culture fermenters fed diets containing two-stage olive cake.

The effects of using effluent bacteria (EB) and solid- (SAB) and liquid- (LAB) associated bacteria and diaminopimelic acid (DAPA) or purine bases (PB) and partially substituting alfalfa hay (AH) by a concentrate including olive cake on the microbial N flow (MNF) and amino acids (AA) flow were investigated with continuous culture fermenters fed AH and a mixture of AH and a concentrate containing barley grains and two-stage olive cake (2:1 ratio) without (AHCO) or with polyethylene glycol (PEG) (AHCOP). The MNF was not different among diets with SAB or LAB (p = 0.302 and 0.203, respectively) and DAPA, but differed with PB (p = 0.021 and 0.014, respectively). With EB both markers detected similar differences, AHCOP showing a higher value (p < 0.05) than AH and AHCO. The MNF was higher (p < 0.001) with PB than DAPA. Daily flow of non-essential AA was not different (p = 0.356) among diets but essential AA flow was higher (p < 0.05) for AH and AHCOP than for AHCO. The SAB presented lower (p < 0.05) total AA than LAB and higher total AA (p < 0.05) for diet AH than AHCO. The AA profile of EB was similar to that of LAB, but alanine and leucine were higher (p < 0.05) in EB than in LAB. Microbial contribution to AA flow was 45.4%, 55.6% and 58.1% for diets AH, AHCO and AHCOP respectively. With both markers, microbial AA flow was higher (p < 0.05) for diet AHCOP compared with AH (451 and 355 mg/day, respectively), but not different (p > 0.05) for AHCOP and AHCO (389 mg/day). The results would indicate that olive cake could be used in the practical feeding of small ruminants without negatively affecting microbial AA N supply.

Effects of concentrate replacement by feed blocks on ruminal fermentation and microbial growth in goats and single-flow continuous-culture fermenters.

The effect of replacing concentrate with 2 different feed blocks (FB) on ruminal fermentation and microbial growth was evaluated in goats and in single-flow continuous-culture fermenters. Diets consisted of alfalfa hay plus concentrate and alfalfa hay plus concentrate with 1 of the 2 studied FB. Three trials were carried out with 6 rumen-fistulated Granadina goats and 3 incubation runs in 6 single-flow continuous-culture fermenters. Experimental treatments were assigned randomly within each run, with 2 repetitions for each diet. At the end of each in vivo trial, the rumen contents were obtained for inoculating the fermenters. For each incubation run, the fermenters were inoculated with ruminal fluid from goats fed the same diet supplied to the corresponding fermenter flask. The average pH values, total and individual VFA, and NH(3)-N concentrations, and acetate:propionate ratios in the rumen of goats were not affected (P >or= 0.10) by diet, whereas the microbial N flow (MNF) and efficiency were affected (P or= 0.05), and total (P=0.02), NH(3) (P=0.005), and non-NH(3) (P=0.02) N flows, whereas the efficiency of VFA production was not affected (P=0.75). The effect of diet on MNF and efficiency depended on the bacterial pellet used as a reference. An effect (P<0.05) of diet on the composition of solid- and liquid-associated bacteria was observed. The compositions of liquid-associated bacteria in the fermenter contents and effluent were similar (P=0.05). Differences (P<0.001) between in vivo and in vitro values for most fermentation variables and bacterial pellet compositions were found. Partial replacement of the concentrate with FB did not greatly compromise carbohydrate fermentation in unproductive goats. However, this was not the case for MNF and efficiency. Differences between the results obtained in vivo and in vitro indicate a need to identify conditions in fermenters that allow better simulation of fermentation, microbial growth, and bacterial pellet composition in vivo. Reduced feeding cost could be achieved with the inclusion of FB in the diets of unproductive goats without altering rumen fermentation.

Comparison of microbial fermentation of high- and low-forage diets in Rusitec, single-flow continuous-culture fermenters and sheep rumen.

Eight Rusitec and eight single-flow continuous-culture fermenters (SFCCF) were used to compare the ruminal fermentation of two diets composed of alfalfa hay and concentrate in proportions of 80 : 20 (F80) and 20 : 80 (F20). Results were validated with those obtained previously in sheep fed the same diets. Rusitec fermenters were fed once daily and SFCCF twice, but liquid dilution rates were similar in both types of fermenters. Mean values of pH over the 12 h postfeeding were higher (P < 0.001) in Rusitec than in SFCCF, with diet F80 showing higher values (P < 0.001) in both types of fermenters. Concentrations of total volatile fatty acids (VFA) were higher (P < 0.001) in SFCCF than in Rusitec, and in both systems were higher (P = 0.002) for diet F20 than for diet F80. There were significant differences between systems in the proportions of the main VFA, and a fermentation system × diet interaction (P < 0.001) was detected for all VFA with the exception of valerate. No differences (P = 0.145) between the two types of fermenters were detected in dry matter (DM) digestibility, but NDF, microbial N flow and its efficiency were higher (P = 0.001) in SFCCF compared to Rusitec. Whereas pH values and VFA concentrations remained fairly stable through the day in both in vitro systems, pH dropped and VFA increased shortly after feeding in sheep rumen reaching the minimum and maximal values, respectively, about 4 h after feeding. Both in vitro systems detected differences between diets similar to those found in sheep for liquid dilution rate, pH values, DM digestibility, microbial N flow and growth efficiency. In contrast, acetate/propionate ratios were lower for diet F20 than for F80 in sheep rumen (2.73 and 3.97) and SFCCF (3.07 and 4.80), but were higher for diet F20 compared to F80 (4.29 and 3.40) in Rusitec, with values considered to be unphysiological for high-concentrate diets. In vivo NDF digestibility was affected (P = 0.017) by diet, but no differences between diets (P > 0.05) were found in any in vitro system. A more precise control of pH in both types of fermenters and a reduction of concentrate retention time in Rusitec could probably improve the simulation of in vivo fermentation.

A comparative study of nutrients utilization, alkaline phosphatase activity and creatinine concentration in the serum of sheep and goats fed diets based on olive leaves.

The aim of this study was to compare, in goats and wethers, the nutritive utilization of diets including olive leaves (OL) and the possible detrimental effect of that by-product. Three different diets were studied: OL, OL plus polyethylene glycol (PEG) (OLP) and OL supplemented with barley [164 g/kg dry matter (DM)] and faba beans (59 g/kg DM) (OLSUP). Apparent digestibility of nutrients and energy and nitrogen balances were determined along with creatinine concentrations and alkaline phosphatase (ALP) activity in the serum. The apparent digestibility of OL was low and similar in goats and wethers (54.4% and 53.5%, 22.2% and 21.6% and, 47.7% and 46.6% for DM, crude protein and neutral detergent fibre in goats and wethers, respectively). The addition of PEG did not improve (p > 0.05) digestibility of OL, although a slightly beneficial effect on the digestion of structural carbohydrates was observed (up to 8.4% and 7.10% in goats and wethers, respectively). The supplementation of OL with barley and faba beans increased (p < 0.001) the apparent digestibility of nutrients and the energy value. The consumption of the OL led to high ALP activity in the serum. Despite the higher sensitivity of wethers to Cu levels, our results show a similar digestive use of OL by wethers and goats and suggest the need of further comparative investigations focusing on the effect of the presence of high Cu levels in the by-product on the animal health.