Influence of condensed tannins from Brazilian semi-arid legumes on ruminal degradability,microbial colonization and ruminal enzymatic activity in Saanen goats

The present study aimed at determining the influence of condensed tannins present in the Brazilian legume species Mimosa hostilis, Mimosa caesalpinifolia and Bauhinia cheilantha on ruminal degradability, microbial colonization and enzymatic activity. Polyethylene glycol (PEG) was used to reduce the astringency and concentration of soluble condensed tannins. Four ruminally-cannulated Saanen goats (60 ± 8 kg BW) were fed, in two experimental periods, with a hay diet based on the studied legumes treated or non-treated with PEG. Voluntary intake, microbial colonization, DM, CP, NDF, and ruminal degradability of PEG treated and non-treated forage leaves, as well as pH, ammonia and 1,4 β-endoglucanase activity of the rumen content were evaluated. Astringency and soluble tannin concentration of the studied legumes were reduced by approximately 70% and 50%, respectively, with PEG treatment. Average DM intake was higher for the treated diet (16.76 g DM/kg BW/day against 13.06 g DM/kg BW/day). Percentile values for degradation parameters and for potential and effective degradabilities of DM, CP and NDF were also affected by the tannins, but at different intensities. Electron microscopic observations of ruminally-incubated legume leaves showed a more effective microbial colonization of PEG-treated leaves for all legume species. A decrease in pH and an increase in ammonia concentration and in endoglucanase activity in the ruminal content was also observed for PEG-treated diets at all sampling periods. Condensed tannins of the studied legume species have influenced the adhesion conditions, colonization and enzymatic activity of the microbial ecosystem, and consequently the ruminal degradation of the different dietary fractions. For this reason, the reduction in condensed tannin would be of great importance to improve the nutrition of ruminant feeding of these species.     

Chemical characterization,in vitro dry matter and ruminal crude protein degradability and microbial protein synthesis of some cowpea (Vigna unguiculata L. Walp) haulm varieties

A study was carried out to evaluate the chemical composition, in vitro apparently and truly degraded dry matter (DM), utilizable crude protein at the duodenum (uCP) (total CP at the duodenum minus endogenous CP), methane production, and short chain fatty acid production of haulms of six cowpea varieties. The study was arranged in a 2 × 2 × 2 factorial design, with three replicates. Three improved (ITA2, ITA6 and ITA8) and three commercial (Oloyin, Peu and Sokoto) cowpea varieties harvested during wet and dry seasons were used for the study. After an initial gas test to evaluate 96 h gas production profiles of haulms with and without polyethylene glycol (PEG), the time to half maximal gas production was calculated and a second incubation conducted with fermentation stopped at substrate specific half time (t_1/2) and 24 h for each substrate. True DM degradability was measured from incubated residues and combined with gas volume to estimate the partitioning factor. Crude protein flow to the duodenum was estimated by combining gas volume with the measured ammonia nitrogen in the incubated fluid. Addition of PEG did not have any effect (P>0.05) on all the variables determined. Interaction between group (improved vs. commercial) and season was observed for CP (P=0.002), lignin (P=0.003) and hemicellulose (P=0.030) contents of the haulms. A group × season interaction was also observed for some of the variables at both substrate specific t_1/2 and 24 h. Commercial cowpea haulms had greater (P=0.002) microbial mass and produced less (P<0.05) methane than the improved cowpea haulms. The improved cowpea haulms were less (P<0.001) degraded in the rumen and as a result ensured greater (P<0.001) amount of uCP. The results validated that cowpea haulm is an important agro-based by-product that is adequate in protein and energy to sustain ruminant animal production in Nigeria and other Sub-Saharan African countries during the extended dry season.     

Effects of plant diversity on microbial biomass and community metabolic profiles in a full-scale constructed wetland

There has been less understanding of relations of microbial community patterns with plant diversity in constructed wetlands. We conducted a single full-scale subsurface vertical flow constructed wetland (SVFCW, 1000 m²) study focusing on domestic wastewater processing. This study measured the size and structure of microbial community using fumigation extraction and BIOLOG Ecoplate? techniques, to examine the effects of macrophyte diversity on microbial communities that are critical in treatment efficiency of constructed wetlands. We also determined the relationship of plant diversity (species richness) with its biomass production under disturbance of the same wastewater supply. Linear regression analysis showed that plant biomass production strongly correlated with plant species richness (R = 0.407, P < 0.001). Increase in plant species richness increased microbial biomass carbon and nitrogen (R = 0.494, P < 0.001; R = 0.465, P < 0.001) and utilization of amino acids on Ecoplates (R = 0.235, P = 0.03), but limited the utilization of amine/amides (R = ?0.338, P = 0.013). Principal components analysis (PCA) showed that the diversity and community-level physiological profiles (CLPP) of microbial community at 168 h of incubation strongly depended on the presence or absence of plant species in the SVFCW system, but not on the species richness. This is the first step toward understanding relations of plant diversity with soil microbial community patterns in constructed wetlands, but the effect of species diversity on microbial community should be further studied.     

Studies on untreated and urea-treated rice straw from three cultivation seasons: 2. Evaluation of straw quality through in vitro gas production and in sacco degradation measurements

Untreated and urea-treated straw and straw fractions of seven rice varieties from three cultivation seasons have been evaluated on their DM, OM loss and degradation characteristics from in sacco disappearance and in vitro gas production measurements. Drying temperatures from 45°C to 100°C did not seem to influence the degradability of urea-treated rice straw, whereas urea-treated straw dried at freezing temperatures (−35°C) gave slightly higher degradability than higher temperatures. Untreated early season rice straw showed higher degradability than straw of middle and later season rice. There was a significant increase in the degradation of straw after urea treatment, and greatest for late and middle season rice straw. On average, urea-treatment of rice straw increased the DM and OM in sacco losses after 48 h of incubation (48 h) by 24.0% and 30.7%, respectively. In order to study the kinetics of the degradation of fibre fractions, the disappearance in sacco was also estimated for the loss of hemicellulose, cellulose and extractable biogenic silica (EBSi). There was a great variation in the content of silica between varieties. Rice straw degradation seemed to be related to the biogenic silica content (acid detergent insoluble silica (ADISi)). Urea treatment increased the extraction of biogenic silica and hence increased the degradation of hemicellulose and cellulose. The improvement in sacco disappearance of cellulose due to urea treatment was 36.8%, 19.5% and 5.3% for late, middle, and early rice straw, respectively. The degradability was higher for the stem than for the leaf blades and leaf sheaths. The response to urea treatment, however, was higher for leaf sheaths and leaf blades than for the stems, evening out differences in degradability. Urea treatment tended to increase the production of acetic acid whereas there was no effect on propionic and butyric acid production.     

Evaluation of a two-stage in vitro technique for estimating digestibility of equine feeds using horse faeces as the source of microbial inoculum

To develop a two-stage in vitro technique that simulates both pre-caecal and hind gut digestion processes, four enzymatic pre-digestion treatments by pepsin and α-amylase (ET0 = control, ET1 = 2 h pepsin + 2 h amylase, ET2 = 2 h pepsin + 4 h amylase, ET3 = 8 h pepsin + 16 h amylase) were tested on oat hay (OH), barley grain (BG) and soybean meal (SBM). Investigated parameters were enzymatic organic matter digestibility (OMDe), and gas production (G48h, G72h) and OM digestibility (OMD) using horse faeces as a source of microbial inoculum.Enzymatic pre-digestion treatments affected (P<0.05) investigated parameters and their ranking differed among feeds. Only OMD of BG and SBM were higher after the pre-digestion treatment. OMD prior to (ET0) and after ET3 application were, successively, 0.357 versus 0.351 (OH), 0.71 versus 0.79 (BG) and 0.70 versus 0.78 (SBM). Net gas production overestimated fermentation potential of non-pre-digested feeds. G72h (ml/g DM) prior to (ET0) and after ET3 application were, successively, 80.3 versus 58.0 (OH), 151.7 versus 30.4 (BG) and 110.6 versus 37.7 (SBM).It was concluded that the enzymatic pre-digestion treatments effects varied among tested feeds, and that the suggested procedure be extended and validated with a large array of feeds of known digestibility values.     

Utility of buckwheat (Fagopyrum esculentum Moench) as feed: Effects of forage and grain on in vitro ruminal fermentation and performance of dairy cows

Buckwheat is of high value in crop rotations and overall agricultural ecology because of strong rooting and intensive flowering properties, but it is rarely cultivated and information on its nutritional value to ruminants is scarce. The contents of net energy for lactation (NEL), as estimated with the Hohenheim Gas Test (n = 3), were 4.3, 4.9 and 7.5 MJ NEL/kg dry matter (DM) for fresh and ensiled whole buckwheat plants and buckwheat grain, respectively. In two experiments with the Rumen Simulation Technique (Rusitec), ruminal fermentation characteristics of buckwheat forages and buckwheat grain (n = 4/diet) were evaluated. In the first experiment, 0, 300 or 600 g/kg of a pure hay diet were replaced by either fresh or ensiled buckwheat to create five diets. Neither form of buckwheat forages had effects on in vitro ruminal degradability and short chain fatty acid (SCFA) concentrations and composition. The use of fresh buckwheat reduced ruminal ammonia concentrations and enhanced estimated microbial N growth efficiency. These differences did not occur with silage, indicating a change in nutritional value by ensiling. Fresh buckwheat reduced the number of bacteria in the incubated fluid, while ensiled buckwheat reduced that of holotrich protozoa. Methane formation was not influenced by the buckwheat forages. In the second experiment, wheat meal (400 g/kg dietary DM), was replaced stepwise (0.5 and 1.0) by buckwheat grain meal. This did not cause differences in parameters of nutrient degradability, relative N efficiency and total amount and composition of SCFA. Holotrich protozoa counts increased, but total gas formation decreased with increasing dietary level of buckwheat grain. In a final experiment, cows yielding about 40 kg milk/day were fed mixed silage-concentrate diets (n = 4). A control diet contained no buckwheat. In a second diet, maize silage was partly substituted by buckwheat silage (98 g/kg dietary DM). In a third group, part of the energy concentrate was substituted by buckwheat grain meal (94 g/kg). There were no effects on feed intake, milk yield and milk composition. Buckwheat proved to be a plant that offers different feeds of a quality sufficient to be considered suitable in ruminant nutrition.     

In vitro evaluation of the fermentation characteristics of the carbohydrate fractions of hulless barley and other cereals in the gastrointestinal tract of pigs

An in vitro model was used to study the fermentation characteristics of carbohydrate fractions of hulless barley (hB), in comparison to hulled barley (HB), hulled oat and oat groats (OG) in the pig intestine. For this purpose, 6 hulless barley cultivars (hB), varying in β-glucan content (36–99 g/kg DM), were compared to 3 HB cultivars, 2 oat groat samples (OG), 3 oat varieties and a reference sample of wheat. The residue of a pepsin–pancreatin hydrolysis was incubated in a buffered mineral solution inoculated with pig faeces. Gas production, proportional to the amount of fermented carbohydrates, was measured for 48 h and kinetics modelled. The fermented solution was subsequently analyzed for microbial production of short-chain fatty acids (SCFA) and ammonia. In vitro dry matter degradability varied according to ingredient (P<0.001). Higher values were observed for OG, ranging from 0.88 to 0.99 as compared to oat, hB and HB, for which degradability ranged from 0.63 to 0.73, 0.68 to 0.80 and 0.69 to 0.71, respectively. A “cereal type” effect (P<0.05) was observed on fermentation kinetics parameters. Total gas production was higher (P<0.05) with hB (224 ml/g DM incubated) than with HB and oat (188 and 55 ml/g DM incubated, respectively). No difference was observed between hB cultivars (P>0.05) for total gas production but differences (P<0.001) were found for lag time and the fractional rate of degradation. Hulless barley cultivar CDC Fibar (waxy starch) and CDC McGwire (normal starch) started to ferment sooner (lag time of 0.7 and 0.9 h, respectively) than SH99250 (high amylose starch; 1.7 h). The fractional rate of degradation was similar in both hB and OG (0.15/h on average), which was higher than that of HB (0.12/h). The production of SCFA was also higher (P<0.05) with hB (6.1 mmol/g DM incubated, on average) than with HB and oat (4.9 and 2.9 mmol/g DM incubated, respectively). Similar trends were found for SCFA production expressed per g fermented carbohydrates, with higher butyrate and lower acetate ratio. In contrast, oat fermentation generated higher (P<0.05) ammonia concentration (1.4 mmol/g DM incubated, on average) than hB (1.0 mmol/g DM incubated). In summary, hulless barleys, irrespective of cultivar type had higher in vitro fermentability and produced more SCFA and less ammonia than hulled barley and oat. Thus, hulless barleys have a better potential to be used in pig nutrition to manipulate the fermentation activity in the intestine of pigs.     

Effects of preservation procedures of rumen inoculum on in vitro microbial diversity and fermentation

Sheep rumen contents were used as inoculum for an in vitro semi-continuous incubation system to study whether preservation method affects microbial fermentation pattern. Rumen fluid was filtered and either used immediately as inoculum (CTL) or dispensed into 110 mm × 16 mm tubes, that were stored refrigerated at 6 °C for 4 h (REF) or frozen at ?20 °C (FRZ), frozen in liquid N (FLN) or added with 0.04 glycerol and frozen in liquid N (FGL) for 48 h. Frozen inocula were thawed at 39 °C for 2 min before use (16 ml per bottle). Two 24 h incubations with four bottles per treatment were completed. The microbial utilisation of added glycerol after thawing in FGL increased total gas production (P<0.05) and 24 h volatile fatty acid (VFA) production (P<0.05), and also increased propionate and butyrate proportions at the expense of acetate. The other freezing inocula (i.e., FLN and FRZ) reduced the rate of gas production (as ml/g dry matter per hour), compared with CTL in the first 2 and 4 h of incubation (P<0.05), but this was compensated by increased fermentation at 8 and 12 h, respectively. Differences in gas production did not manifest a different VFA pattern at either 6 or 24 h incubation. Bacterial diversity was slightly affected by the preservation process, and the similarity index between untreated inocula and the 24 h incubated CTL samples was 0.690–0.724. Similarity between bacterial communities in FRZ and FLN with that in CTL after incubation was 0.678. The freezing preservation method of rumen inocula for subsequent in vitro gas production studies does not affect microbial fermentation pattern or bacterial biodiversity, provided that processing is rapid enough by using a high surface to volume ratio. Freezing in liquid N is more appropriate than at ?20 °C.     

Variability of soil carbon sequestration capability and microbial activity of different types of salt marsh soils at Chongming Dongtan

Variations in the soil carbon sequestration capability of different types of salt marsh soils at Chongming Dongtan and its influencing factors were studied by analyzing the soil organic carbon (SOC) content, organic matter input and microbial activities. The results indicated that the total SOC content at Area A (southeast of Dongtan, sandy soil with Phragmites communis) was only 46.11% of that of Area B (northeast of Dongtan, clay soil with mixed P. communis and Spartina alterniflora) (P = 0.000 < 0.05), but their organic matter input per year was almost identical. These findings implied that Area B had a lower output of SOC. The microbial biomass at Area A was 3.83 times greater than that at Area B (P = 0.049 < 0.05); the soil catalase and invertase activities at Area A, which were related to carbon metabolism, were 60.31% (P = 0.006 < 0.05) and 34.33% (P = 0.021 < 0.05) higher than at Area B, respectively; and the soil respiration at Area A was also higher than at Area B. These findings implied that the microbial activities at Area A were greater than those at Area B, and therefore the carbon metabolism was rapid, resulting in increased SOC output at Area A. Increased water content and salinity in the clay soil at Area B may inhibit the microbial activities, thereby reducing the decomposition of the organic matter and enhancing carbon sequestration. In addition, some artificial measures for controlling spread of S. alterniflora at Area B (mowing/digging and tillage (M + D); mowing/digging and tillage/waterlogging (M + D + W)) were found to generally improve the microbial activity of soil, thereby increasing SOC output. However, when the two different physical controlling modes were compared, the SOC and microbial activities of the soil subjected to the M + D + W treatment were relatively high and low, respectively, due to waterlogging restraining the microbial metabolism. These findings indicated that the difference in microbial activities was the important factor leading to variability in the SOC sequestration capability between Areas A and B. Additionally, with the exception of soil texture and vegetation types, environmental conditions and artificial turbulence also influenced microbial activities of soil, and hence SOC output and organic carbon sequestration capability.     

Effects of untreated and heat-treated canola presscake on milk yield and composition of dairy cows

Five primiparous and five multiparous Holstein cows were used in two Latin square design experiments to determine effects of feeding unheated and heated canola presscake on milk yield and composition, and milk fatty acid concentrations of lactating dairy cows. Five diets that differed in level and source of dietary fat were formulated: a low fat control diet with 30 g kg⁻¹ fat from tallow, an unheated canola presscake supplemented diet (50 g kg⁻¹ fat), a heated canola presscake supplemented diet (50 g kg⁻¹ fat), a high tallow plus unheated canola meal supplemented diet (50 g kg⁻¹ fat), and a high tallow plus heated canola meal supplemented diet (50 g kg⁻¹ fat). In sacco ruminal degradability of heated and unheated canola presscake was compared with that of heated and unheated canola meal in a randomized complete block design using two ruminally fistulated cows. Heat treatment reduced ruminal DM and CP degradability of canola presscake. Multiparous cows fed diets supplemented with heated or unheated canola presscake produced more milk than those fed diets containing similar levels of fat from tallow with heated or unheated canola meal, respectively. High levels of fat from any diet reduced milk fat percentage for cows of either parity. Feeding heated canola products increased milk and milk protein yields in primiparous cows only, but cows of both parities fed diets containing canola presscake produced milk with lower concentrations of C12:0, C14:0, and C16:0 fatty acids than cows fed the canola meal and tallow diets, although concentrations of C18:1 n-9 were unaffected by fat source or level. Feeding canola products to dairy cows can alter milk fatty acid profile, but only primiparous cows have increased productivity as a result of feeding heated, versus unheated, canola presscake.     

Medium optimization for the high yield production of single (+)-terrein by Aspergillus terreus strain PF26 derived from marine sponge Phakellia fusca

Terrein has potential application in the fields of medicine, cosmetology and agriculture, however, the chemical synthesis of terrein with single configuration is a difficult task, and the biosynthesis of terrein always results in low production (ca. 0.33–400 mg/L). In this study, we reported an Aspergillus terreus strain PF26 which could produce (+)-terrein on a high level. After the selection of a suitable basic medium, the component concentrations were optimized using Plackett–Burman design and response surface methodology. Consequently, an optimal medium containing 28.41 g glucose, 23.18 g maltose, 20.00 g mannitol, 8.52 g malt extract, 10.00 g monosodium glutamate 10.00 g NH₄Cl in 1 L ASW was obtained, and a high (+)-terrein production of 3.71 g/L fermentation broth was achieved, which represents the highest fermentation production of (+)-terrein to date. The result highlighted the industry's potential of A. terreus strain PF26 in the production of bioactive (+)-terrein on a large-scale.     

Antagonist mechanical contribution to resultant maximal torque at the ankle joint in young and older men

A recorded muscular torque at one joint is a resultant torque corresponding to the participation of both agonist and antagonist muscles. This study aimed to examine the effect of aging on the mechanical contributions of both plantar- and dorsi-flexors to the resultant maximal voluntary contraction (MVC) torques exerted at the ankle joint, in dorsi-flexion (DF) and plantar-flexion (PF). The estimation of isometric agonist and antagonist torques by means of an EMG biofeedback technique was made with nine young (mean age 24 years) and nine older (mean age 80 years) men. While there was a non-significant age-related decline in the measured resultant DF MVC torque (?15%; p = 0.06), there was a clear decrease in the estimated agonist MVC torque exerted by the dorsi-flexors (?39%; p = 0.001). The DF-to-PF resultant MVC torque ratio was significantly lower in young than in older men (0.25 vs. 0.31; p = 0.006), whereas the DF-to-PF agonist MVC torque ratio was no longer different between the two populations (0.38 vs. 0.35; p > 0.05). Thus, agonist MVC torques in PF and DF would be similarly affected by aging, which could not be deduced when only resultant torques were examined.     

The degradation of coniferyl alcohol and the complementary production of chlorogenic acids in the growth culture of Streptomyces albogriseolus KF977548 isolated from decaying wood residues

Coniferyl alcohol is one of the major precursors of lignin; the most abundant aromatic compound and a natural resource currently receiving attention because of the value-added metabolites resulting from its degradation. Growth study of Streptomyces albogriseolus KF977548 (strain AOB) isolated from decaying wood residues in a tropical estuarine ecosystem was carried out using coniferyl alcohol as a sole carbon source. Cell growth and metabolite production were monitored at 24 h interval by dry weight measurements and HPLC, LC–MS-DAD analyses. Biochemical and PCR assays were carried out to detect the major catabolic enzymes of interest. Strain AOB utilized coniferyl alcohol completely within 72 h (μ = 0.204 h⁻¹, T_d = 3.4 h). Laccase and peroxidase were released into the growth medium up to 0.099 and 98 μmol/mL respectively. Protocatechuate 3, 4-dioxygenase and demethylase were detected in the genome whilst ortho-adipate pathway was clearly indicated. Growth on coniferyl alcohol or caffeic acid as mono substrates resulted in the production of secondary metabolites identified by HPLC–MS as 1-caffeoylquinic and 3,4,5-tricaffeoylquinic acids, known as chlorogenic acids, in the culture medium. The microbial production of chlorogenic acids from a lignin-related substrate base by strain AOB could arouse a plausible biotechnological process.     

Effects of zinc deficiency and zinc supplementation on homocysteine levels and related enzyme expression in rats

Methionine synthase (MS) and betaine-homocysteine methyltransferase (BHMT) are both zinc (Zn)-dependent methyltransferases and involved in the methylation of homocysteine. The objective of this study was to investigate the effects of dietary Zn supply on homocysteine levels and expression of the two enzymes in growing rats. Male weanling Sprague-Dawley rats were assigned randomly to four dietary groups (n = 8/group) for 3 weeks: Zn deficient (ZD; <1 mg Zn/kg); Zn control (ZC; 30 mg Zn/kg); Zn supplemented (ZS; 300 mg Zn/kg); pair fed (PF; 30 mg Zn/kg) to the ZD group. Serum and femur Zn concentrations were 83% and 58% lower in ZD, and 49% and 62% higher in ZS compared to ZC (P < 0.001), respectively. The ZD rats had lower feed intake (37%), body weight gains (45%), liver (43%) and kidney (31%) weights than those of ZC (P < 0.001), but these parameters in ZD were not significantly different from the PF controls. Serum homocysteine concentrations were 65% higher in ZD compared to PF (P < 0.05), and there was no significant difference in serum folate levels between ZD and PF groups. The mRNA expression of liver and kidney MS was 57% and 38% lower in ZD than PF (P < 0.001), respectively. Hepatic and renal BHMT mRNA levels were not altered in ZD compared to controls. The aforementioned measurements were not significantly different between ZS and ZC groups, except Zn levels. These results demonstrated that homocysteine homeostasis appeared to be disturbed by Zn deficiency but not Zn supplementation, and elevated serum homocysteine might be due to reduced expression of MS during Zn deficiency.     

Kinetic modeling and implementation of superior process strategies for β-galactosidase production during submerged fermentation in a stirred tank bioreactor

This paper reports development and implementation of superior fermentation strategies for β-galactosidase production by Lactobacillus acidophilus in a stirred-tank bioreactor. Process parameters (aeration and agitation) were optimized for the process by application of Central Composite Design. Aeration rate of 0.5 vvm and agitation speed of 250 rpm were most suitable for β-galactosidase production (2001.2 U/L). Further improvement of the operation in pH controlled environment resulted in 2135 U/L of β-galactosidase with productivity of 142.39 U/L h. Kinetic modeling for biomass and enzyme production and substrate utilization were carried out at the aforementioned pH controlled conditions. The logistic regression model (X₀ = 0.01 g/L; X_max = 2.948 g/L; μ_max = 0.59/h; R² = 0.97) was used for mathematical interpretation of biomass production. Mercier's model proved to be better than Luedeking–Piret model in describing β-galactosidase production (P₀ = 0.7942 U/L; P_max = 2169.3 U/L; P_r = 0.696/h; R² = 0.99) whereas the latter was more efficient in mathematical illustration of lactose utilization (m = 0.187 g/g h; Y_x/s = 0.301 g/L; R² = 0.98) among the two used in this study. Strategies like fed-batch fermentation (3694.6 U/L) and semi-continuous fermentation (5551.9 U/L) further enhanced β-galactosidase production by 1.8 and 2.8 fold respectively.     

Lactobacillus plantarum effects on silage fermentation and in vitro microbial yield

In four parallel experiments, herbage [three harvests of alfalfa (308 to 379 g dry matter (DM)/kg), one of whole-plant corn (331 g DM/kg)] was ensiled with three different treatments: no inoculant (control), Lactobacillus plantarum (LP) or formic acid (FA), in 1-L mini-silos and fermented for 60 d at room temperature (22 °C). Mini-silos were opened and analyzed for fermentation characteristics and soluble N fractions. A subsample of wet silage from each mini-silo was ground to 4 mm and stored at ?20 °C. Silages were thawed and subjected to 9 h ruminal in vitro incubations to measure gas production and volatile fatty acid (VFA) production as well as microbial biomass yield (MBY) and microbial non-ammonia N (MNAN) formation using ¹⁵N as a marker. In all four experiments, silage fermentation products and pH indicated good preservation across all treatments. Analysis of data showed that FA- and LP-treated silages had lower concentrations of ammonia-N and free amino acids N than control. The FA treatment was lower in soluble N, but higher in peptide-N, than control. Silage pH was lowest in FA (4.25), followed by LP (4.28), and control (4.38). Ruminal in vitro gas production and VFA concentrations were not different among treatments (P>0.05). Compared to control, FA- and LP-treated silage yielded greater MNAN and MBY. These findings suggested that L. plantarum preserved more true protein during silage fermentation than control, which in turn increased in vitro ruminal microbial growth.     

Effects of plant diversity on biomass production and substrate nitrogen in a subsurface vertical flow constructed wetland

Most biodiversity experiments have been conducted in grassland ecosystems with nitrogen limitation, while little research has been conducted on relationships between plant biomass production, substrate nitrogen retention and plant diversity in wetlands with continuous nitrogen supply. We conducted a plant diversity experiment in a subsurface vertical flow constructed wetland for treating domestic wastewater in southeastern China. Plant aboveground biomass production ranged from 20 to 3121 g m^?2 yr^?1 across all plant communities. In general, plant biomass production was positively correlated with species richness (P = 0.001) and functional group richness (P = 0.001). Substrate nitrate concentration increased significantly with increasing plant species richness (P = 0.046), but not with functional group richness (P = 0.550). Furthermore, legumes did not affect biomass production (P = 0.255), retention of substrate nitrate (P = 0.280) and ammonium (P = 0.269). Compared to the most productive of the corresponding monocultures, transgressive overyielding of mixed plant communities did not occur in most polycultures. Because greater diversity of plant community led to higher biomass production and substrate nitrogen retention, thus we recommend that plant biodiversity should be incorporated in constructed wetlands to improve wastewater treatment efficiency.     

Dependence of forage quality and methanogenic potential of tropical plants on their phenolic fractions as determined by principal component analysis

This study was conducted to elucidate relationships among various phenolic fractions in, and methane (CH₄) emissions from, tropical plants when incubated in ruminal fluid in vitro. As a second objective, principal component analysis (PCA) was tested for its utility in screening plants for their ability to reduce CH₄ formation at simultaneously acceptable nutritional quality. Leaves from 27 tropical plants were analyzed for their nutritional composition and various phenolic fractions. They were incubated in vitro using the Hohenheim gas test method. Variables measured after 24 h of incubation were total gas and CH₄ production, and pH, ammonia, bacterial and protozoal counts, as well as short-chain fatty acids in the incubation fluid. In vitro organic matter (OM) digestibility was computed by a standard equation. The data obtained was subjected to analysis of variance, correlation, regression and PCA. Among phenolic fractions, total phenols had the closest relationship with CH₄/digestible OM (r = ?0.84, P<0.001). The total tannin fraction contributed strongly to this effect (r = ?0.74, P<0.001) whereas the non-tannin phenol fraction was less important (r = ?0.45, P<0.05). Methane reduction by the influence of non-tannin phenols was not associated with a negative effect on protein degradation, while this was the case with tannins. Condensed (r = ?0.60, P<0.01) and hydrolysable tannins (r = ?0.60, P<0.01) contributed to the decrease in CH₄/digestible OM. The loading plot of PCA showed that dietary crude protein (CP) content and incubation fluid ammonia, total short-chain fatty acids, propionate, valerate, iso-butyrate, iso-valerate as well as in vitro OM digestibility were clustered. They had inverse directions to contents of fiber fractions and incubation fluid acetate proportion and acetate-to-propionate ratio. The methane-to-total-gas ratio had the opposite effect of the contents of any phenolic fraction. Plants possessing a favorable forage quality, based on the corresponding PCA score plot, were Carica papaya, Manihot esculenta, Morinda citrifolia, Sesbania grandiflora and Melia azedarach, whereas CH₄ mitigating plants included Swietenia mahagoni, Acacia villosa, Eugenia aquea, Myristica fragrans and Clidemia hirta. All phenolic fractions studied reduced CH₄ emissions from in vitro incubations with ruminal fluid and PCA seems useful to screen plants for high nutritional quality and low ruminal CH₄ formation. However, high forage quality seemed to be partially associated with high CH₄ emission. The search for plants rich in non-tannin phenols might be promising as these compounds appear to decrease CH₄ while they obviously have less negative effect on protein degradation as compared to the tannin fractions.     

Feeding soybean meal increases the blood level of isoflavones and reduces the steroidogenic capacity in bovine corpora lutea,without affecting peripheral progesterone concentrations

Thirty-three Holstein-Friesian cows were followed from 14 days pre partum until the fourth ovulation post partum. Housing conditions and basic ration were identical for all animals. Concentrates were individually supplemented according to the daily milk production level, using two different types of protein rich concentrates: soybean meal and rapeseed meal. Soybean and rapeseed meal are known to be respectively high and low in isoflavones. Cows were randomly divided into three groups and blocked for parity. Group I (n = 11) was supplemented with soybean meal and acted as control group. Groups II (n = 11) and III (n = 11) were respectively supplemented with soybean and rapeseed meal and were subjected to a biopsy sampling of the corpus luteum at day 9 of the first three postpartal estrous cycles.Soybean meal supplementation to lactating dairy cows (1.72 kg on average) induced an increase in the blood concentration of equol, dihydrodaidzein, o-desmethylangolensin in both soy groups and resulted in a reduced area occupied by steroidogenic (P = 0.012) and endothelial cells (P = 0.0007) in the luteal biopsies. Blood concentrations of equol and glycitein were negatively correlated with the areas occupied by steroidogenic (r = −0.410 with P = 0.0002, respectively r = −0.351 with P = 0.008) and endothelial cells (r = −0.337 with P = 0.01, respectively r = −0.233 with P = 0.085) in the 3 first estrous cycles. The latter however did not affect the diestrous peripheral blood progesterone concentration.     

Spatial and temporal changes in the microbial community in an anaerobic swine waste treatment lagoon

Microorganisms are central to both the beneficial (organic degradation, nutrient removal, biogas production) and detrimental (odor production, pathogen contamination) effects of swine waste storage systems. In this study, both quantitative (real-time polymerase chain reaction) and qualitative (denaturing gradient gel electrophoresis, cloning, sequence analysis) molecular analyses were used to track spatial and temporal changes in the microbial community of swine slurry from a 0.4 ha anaerobic lagoon. The lagoon, located in a region of western Kentucky which has a humid, subtropical environment, was sampled on a monthly basis (n = 10) over a period of one year at four different depths (top, 51 cm from the top, 152 cm from the top, and bottom >198 cm). The concentration and diversity of Bacteroides sp. was seasonal (up to 90% decrease between March and June). Hespellia sp. and other clostridial species, on the other hand, were endemic in the slurry (concentrations up to 1.0 × 10⁷ cells mL^?1 slurry) regardless of time of the year or lagoon depth. Results suggest that there were seasonal effects on the microbial community in the swine lagoon, while the effect of depth was not as pronounced. Seasonal changes in the microbial community in stored wastes may be (directly or indirectly) correlated with changes in malodor emissions from lagoons.