Effect of enzyme extracts isolated from white-rot fungi on chemical composition and in vitro digestibility of wheat straw

A series of in vitro experiments were completed to evaluate the potential of enzyme extracts, obtained from the white-rot fungi Trametes versicolor (TV1, TV2), Bjerkandera adusta (BA) and Fomes fomentarius (FF), to increase degradation of cell wall components of wheat straw. The studies were conducted as a completely randomized design and analysed using one-way ANOVA. Enzyme activities of the extracts, previously obtained from a liquid culture medium, were characterized in terms of laccase and peroxidase for ligninolytic activity. Carboxymethyl cellulase (CMCase) and avicell digesting cellulase (Avicelase) were used for cellulolytic enzyme assays. Wheat straw samples were incubated with enzyme extracts in a citrate buffer (pH 5.0) in a forced air oven at 25 °C for 6 days. In vitro NDF digestibility (IVNDFD), and the rate and extent of NDF fermentation, without and after incubation with the white-rot enzyme extracts, were determined using a gravimetric microbiological method and a gas production technique, respectively. Results from cell wall chemical composition showed that TV2 and BA enzyme extracts decreased NDF concentration (P<0.05) and that TV1 had higher activity (P<0.05) towards cellulose. There was an increase in IVNDFD (P<0.05), resulting from treatment of wheat straw with enzyme extracts from BA, TV1 and TV2, reaching a difference of 13% for TV2 (P<0.05), versus the non-treated straw control. Treatment with enzyme extract from TV2 caused increased gas production (P<0.05) after the first 20 h of incubation, and also increased the maximum rate of gas production, thus enhancing fermentation kinetics. This study indicates that enzyme extracts from white-rot fungi can be used to develop new approaches to overcome low digestibility of some plant cell walls. Utilization of different substrates to produce enzyme extracts can lead to production of viable ligninolytic complexes which could improve the nutritive value of fibrous feeds.     

The effect of latitudinal gradient on the species diversity of Chinese litter-dwelling thrips

To understand the global distribution patterns of litter-dwelling thrips, a total 150 leaf litter samples were collected from 6 natural reserves located in three climatic regions, temperate, subtropical and tropical. The results showed the relative abundance of Thysanoptera was over 3.0% in 4 natural reserves from subtropical and tropical zone, and reached 5.9% in one tropical reserve, only less than Acarina and Collembola. In contrast it was only 0.3% in the warm temperate natural reserves, and no thrips were collected in a mid temperate reserve. The order on the average species numbers per plot of litter thrips was tropic > subtropics > temperate (n=25, p<0.05). Mean density of litter thrips per plots in the tropics and subtropics was significantly higher than that in the temperate region (n=25, p<0.05), but the average density was not significantly different between tropical and subtropical zones (n=25, p>0.05). The diversity of litter thrips in the tropics and subtropics was much higher than that in the temperate area based on comparsions of Shannon-Wiener diversity index (H’), Pielou eveness index (J), and Simpson dominance index (D). All of these results indicated that litter-dwelling thrips lived mainly in tropical and subtropical regions; meanwhile, species number and relative abundance increased with decreasing latitude.     

The potential of Commelina benghalensis as a forage for ruminants

To evaluate the potential of Commelina benghalensis as a forage for ruminants, effects of plant maturity on chemical composition, rumen degradability as well as its increased dietary inclusion level on intake, digestibility and N balance in sheep fed Sorghum almum were investigated with forage obtained from the wild, re-established and harvested at 6, 10 and 14 weeks of growth. Composite herbage samples were analyzed for dry matter (DM), chemical components, total extractable phenolics (TEP) and amino acid content. In sacco rumen degradability measurements used six individually confined wethers (8 ± 0.5 months of age; 21 ± 2.6 kg live-weight (LW)) fitted with rumen cannulae and fed a ration of 3:1 fresh S. almum and Medicago sativa hay (about 1:2 on a DM basis). In sacco bags containing 5 g each of dry herbage were inserted into the rumen and withdrawn sequentially after 0, 12, 24, 36 and 48 h. Metabolizable energy (ME) was estimated from 24 h in vitro gas production. In the digestibility study, 12 wethers fitted with rumen canula were housed in metabolic crates and allotted to four treatment diets (i. e., D0, D10, D20 and D30) constituted from fresh S. almum and pre-wilted C. benghalensis in a randomized complete block design. The control diet (D0) was 3 kg fresh S. almum (≈535.5 g DM/wether/d about 30 g/kg LW), whereas D10, D20 and D30 were D0 +300, 600 or 900 g of wilted C. benghalensis (≈34, 68 or about 102 g DM/wether/d), respectively. The study lasted for 21 d. Dry matter, fibre and TEP content increased (P<0.001) with maturity of the forage, whereas those of CP and EE decreased (P<0.0001) over the same period. Amino acids (AA) also declined with maturity (P<0.05). Rumen degradability of DM and OM were unaffected, but DM intake increased linearly (P<0.0001) at a decreasing rate (Q: P<0.05) and DM digestibility (DMD) and N intake increased linearly (P<0.01 and P<0.0001, respectively) as level of C. benghalensis in the diet increased. Results indicate that advancing maturity affected chemical composition, but not rumen degradability, of C. benghalensis and also demonstrated that inclusion of C. benghalensis in S. almum diet improved intake, digestibility and N intake, suggesting its potential use as a feed supplement.     

Influence of the maize silage to grass silage ratio and feed particle size of rations for ruminants on the community structure of ruminal Firmicutes in vitro

Aims: To investigate the effect of the forage source and feed particle size (FPS) in ruminant rations on the composition of the ruminal Firmicutes community in vitro. Methods and Results: Three diets, varying in maize silage to grass silage ratio and FPS, were incubated in a rumen simulation system. Microbial samples were taken from the liquid fermenter effluents. Microbial community analysis was performed by 16S rRNA‐based techniques. Clostridia‐specific single‐strand conformation polymorphism profiles revealed changes of the community structure in dependence on both factors tested. The coarse grass silage–containing diets seemed to enhance the occurrence of different Roseburia species. As detected by real‐time quantitative PCR, Ruminococcus albus showed a higher abundance with decreasing FPS. A slightly lower proportion of Bacilli was found with increasing grass silage to maize silage ratio by fluorescence in situ hybridization (FISH). In contrast, a slightly higher proportion of bacterial species belonging to the Clostridium‐clusters XIV a and b was detected by FISH with increasing grass silage contents in the diet. Conclusions: The ruminal Firmicutes community is affected by the choice of the forage source and FPS. Significance and Impact of the Study: This study supplies fundamental knowledge about the response of ruminal microbial communities to changing diets. Moreover, the data suggest a standardization of grinding of feeds for in vitro studies to facilitate the comparison of results of different laboratories.     

Spatial and Temporal Biogeography of Soil Microbial Communities in Arid and Semiarid Regions

Microbial communities in soils may change in accordance with distance, season, climate, soil texture and other environmental parameters. Microbial diversity patterns have been extensively surveyed in temperate regions, but few such studies attempted to address them with respect to spatial and temporal scales and their correlations to environmental factors, especially in arid ecosystems. In order to fill this gap on a regional scale, the molecular fingerprints and abundance of three taxonomic groups – Bacteria, α-Proteobacteria and Actinobacteria – were sampled from soils 0.5–100 km apart in arid, semi-arid, dry Mediterranean and shoreline Mediterranean regions in Israel. Additionally, on a local scale, the molecular fingerprints of three taxonomic groups – Bacteria, Archaea and Fungi – were sampled from soils 1 cm–500 m apart in the semi-arid region, in both summer and winter. Fingerprints of the Bacteria differentiated between all regions (P<0.02), while those of the α-Proteobacteria differentiated between some of the regions (0.01<P<0.09), and actinobacterial fingerprints were similar among all regions (P>0.05). Locally, fingerprints of archaea and fungi did not display distance-decay relationships (P>0.13), that is, the dissimilarity between communities did not increase with geographic distance. Neither was this phenomenon evident in bacterial samples in summer (P>0.24); in winter, however, differences between bacterial communities significantly increased as the geographic distances between them grew (P<0.01). Microbial community structures, as well as microbial abundance, were both significantly correlated to precipitation and soil characteristics: texture, organic matter and water content (R²>0.60, P<0.01). We conclude that on the whole, microbial biogeography in arid and semi-arid soils in Israel is determined more by specific environmental factors than geographic distances and spatial distribution patterns.     

Nanomechanics of the Endothelial Glycocalyx in Experimental Sepsis

The endothelial glycocalyx (eGC), a carbohydrate-rich layer lining the luminal side of the endothelium, regulates vascular adhesiveness and permeability. Although central to the pathophysiology of vascular barrier dysfunction in sepsis, glycocalyx damage has been generally understudied, in part because of the aberrancy of in vitro preparations and its degradation during tissue handling. The aim of this study was to analyze inflammation-induced damage of the eGC on living endothelial cells by atomic-force microscopy (AFM) nanoindentation technique. AFM revealed the existence of a mature eGC on the luminal endothelial surface of freshly isolated rodent aorta preparations ex vivo, as well as on cultured human pulmonary microvascular endothelial cells (HPMEC) in vitro. AFM detected a marked reduction in glycocalyx thickness (266 ± 12 vs. 137 ± 17 nm, P<0.0001) and stiffness (0.34 ± 0.03 vs. 0.21 ± 0.01 pN/mn, P<0.0001) in septic mice (1 mg E. coli lipopolysaccharides (LPS)/kg BW i.p.) compared to controls. Corresponding in vitro experiments revealed that sepsis-associated mediators, such as thrombin, LPS or Tumor Necrosis Factor-α alone were sufficient to rapidly decrease eGC thickness (-50%, all P<0.0001) and stiffness (-20% P<0.0001) on HPMEC. In summary, AFM nanoindentation is a promising novel approach to uncover mechanisms involved in deterioration and refurbishment of the eGC in sepsis.     

An evolutionary-ecological model of the evolution of migratory behavior in the Monarch Butterfly,and its absence in the Queen Butterfly

This paper presents a model that generates testable hypotheses concerning the evolution of long-range migratory behavior in the Monarch Butterfly,Danaus plexippus and the general absence of such behavior in a related form, the Queen,D. gilippus (Lepidoptera: Nymphalidae: Danainae). An attempt is made to reconstruct a probable transition within the Neotropical forest-dwelling danaines associated with woody Asclepiadaceae (and to a lesser extent, perhaps some Apocyanaceae) as larval food plants to a progenitor stock of the Monarch and Queen in more seasonal tropical regions, and eventually temperate regions, and associated with herbaceous asclepiads such asAsclepias. A basic premise of the proposed hypothetical model is that the colonization of secondary habitats in seasonal tropical regions of Central America preadapted both forms to an eventual colonization of the sub-temperate and temperate zone of North America. The assumed evolutionary diversification of the herbaceousAsclepias species in North America provided an evolutionary stepping stone for the expansion of these danaines into this region. Owing to strong selection arising from the co-association of the Monarch and Queen with the same species ofAsclepias in the subtropical region of North America, eventually there was selection for the colonization of the higher latitudes by the Monarch, whereAsclepias thrived. Being essentially a tropical insect, the Monarch evolved an obligatory long-range migratory behavior to allow colonization of the temperate zone annually, and necessitating the use of overwintering sites in Mexico and other places.     

Implementation and Monitoring of Soil Bioengineering Measures at a Landslide in the Middle Mountains of Nepal

Legume tissue quality is a key factor for enhancement of feed resources and contribution to soil fertility in mixed crop-livestock production systems. To compare methods used by soil scientists and animal-nutritionists to assess quality of plant materials, three woody tropical legumes with contrasting qualities were used: Indigofera zollingeriana Miq. (Indigofera), Cratylia argentea Benth. (Cratylia) and Calliandra houstoniana (Mill.) Stan. var. calothyrsus (Meiss.) Barn. CIAT 20400 (Calliandra). Plant material of each legume was used either fresh, freeze-dried, frozen, oven-dried (60 °C) or air-dried in order to estimate extents and rates of aerobic degradation in litterbags on the soil during 140 days and anaerobic degradation in an in-vitro gas production experiment during 144 h. Results showed, that aerobic decomposition rates of leaf tissues were highest for Indigofera (k = 0.013 day⁻¹), followed by Cratylia (k = 0.004 day⁻¹) and Calliandra (k = 0.002 day⁻¹). Gas production rates evaluated under anaerobic conditions, were highest for Indigofera (k = 0.086 h⁻¹), intermediate for Cratylia (k = 0.062 h⁻¹) and lowest for Calliandra (k = 0.025 h⁻¹). Decomposition and gas production rates differed (P < 0.001) among species. Differences between post harvest treatments were not statistically significant (P > 0.05). The extent of decomposition was highest for Indigofera (82.5%, w/w), followed by Cratylia (44.6%) and Calliandra (26.4%). The extent of gas production was highest for Indigofera (218.8 ml), followed by Cratylia (170.1ml) and Calliandra (80.1 ml). Extent of decomposition and extent of gas production were significantly different (P < 0.001) among species. In contrast to the extent of decomposition, the extent of gas production was affected (P < 0.001) by sample post harvest treatments. Highest gas production was observed for the fresh and frozen treatments. The forage quality parameters that best correlated with aerobic and anaerobic degradation were lignin+bound condensed tannins, lignin+total condensed tannins/N, indigestible acid detergent fibre (IADF) and in-vitro dry matter digestibility (IVDMD). Results showed that differences in decomposition and digestibility were more related to intrinsic plant quality parameters than to changes in tissue quality induced by post harvest treatments. In addition, we found that rate of aerobic degradation of legume leaves on the soil was highly correlated (r > 0.80, P < 0.001) to IVDMD and gas production (r = 0.53, P < 0.001). These results indicate that plant measurements (IADF, IVDMD and gas production) used to assess forage quality in animal nutrition studies are more rapid and resource saving predictors for aerobic decomposition of tropical legumes than initial plant quality ratios (lignin+polyphenols/N and lignin+total condensed tannins/N) commonly used by many researchers. Furthermore, this study confirms the potential usefulness of IVDMD for screening tropical legumes for soil fertility management.     

Effects of Fermenten supplementation to beef cattle

Two experiments were conducted to evaluate a commercially available supplemental N source for beef cattle (Fermenten^®; Church & Dwight Co., Inc., Princeton, NJ, USA). The first experiment evaluated kinetics of in vitro NH₃-N release using batch cultures of rumen fluid incubated with: control (no N added), soybean meal, urea, and Fermenten^®. Ammonia-N was measured at 0, 0.5, 2, 4, 6, 8, 12 and 24 h after incubation began. A treatment by time interaction (P<0.01) occurred in which, during the initial 2 h, Fermenten^® cultures had the highest (P<0.01) NH₃-N but, from 4 to 24 h, the highest (P<0.01) NH₃-N concentrations were with urea-incubated cultures. The total increase in NH₃-N concentrations from 0 to 24 h of incubation was less for Fermenten^® (P<0.01) than for the soybean meal and urea. The second experiment assessed effects of Fermenten^® supplementation on growth, blood parameters, voluntary forage intake and reproductive performance of beef heifers. Sixty heifers, stratified by initial body weight (BW), were randomly allocated to one of two treatments that consisted of iso-nitrogenous grain-based supplements containing either Fermenten^® (72 g/kg, as-fed) or urea (9.7 g/kg, as-fed). Supplements were offered three times weekly at a rate of 2.4 kg of dry matter per heifer daily. Shrunk BW was measured on days 0 and 112 for calculation of daily body weight gain. Body volume measurements were completed on days 0, 28, 56, 84 and 112, whereas pelvic area was assessed on days 0, 56 and 112. Blood samples were collected on days 28, 56, 84 and 112 for analysis of metabolites and hormones. On day 56, 2 heifers, which were randomly selected from each pasture, were placed in individual feeding stations for 26 days to determine treatment effects on voluntary forage intake. On day 112, all heifers were grouped by treatment and exposed to bulls for 60 days. Fewer heifers offered the Fermenten^® supplement attained puberty (P<0.05) and became pregnant during the study compared to heifers fed urea (0.60 and 0.93, respectively; P<0.01). Addition of Fermenten^® to batch cultures of rumen fluid rapidly increased NH₃-N concentrations, whereas further increases occurred in a slower and steady rate. Beef heifers fed a supplement containing Fermenten^® had similar growth and development, but inferior reproductive performance, than heifers fed a supplement containing urea.     

Early-Life Gut Bacteria Associate with IL-4?, IL-10? and IFN-γ Production at Two Years of Age

Microbial exposure early in life influences immune maturation and potentially also the development of immune-mediated disease. Here we studied early-life gut colonization in relation to cytokine responses at two years of age. Fecal samples were collected from infants during the first two months of life. DNA was extracted from the fecal samples and Bifidobacterium (B.) adolescentis, B. breve, B. bifidum, a group of lactobacilli (L. casei, L. paracasei and L. rhamnosus) as well as Staphylococcus (S.) aureus were detected with real time PCR. Peripheral mononuclear cells were stimulated with phytohaemagglutinin (PHA) and numbers of IL-4−, IL-10− and IFN-γ secreting cells were evaluated using ELISpot. We further stimulated peripheral blood mononuclear cells with bacterial supernatants in vitro and assessed the IL-4−, IL-10− and IFN-γ inducing capacity by flow cytometry and ELISA. Early S. aureus colonization associated with higher numbers of IL-4− (p = 0.022) and IL-10 (p = 0.016) producing cells at two years of age. In contrast to colonization with S. aureus alone, co-colonization with lactobacilli associated with suppression of IL-4− (p = 0.004), IL-10− (p = 0.004) and IFN-γ (p = 0.034) secreting cells. In vitro stimulations of mononuclear cells with bacterial supernatants supported a suppressive role of L. rhamnosus GG on S. aureus-induced cytokine responses. We demonstrate that the early gut colonization pattern associates with the PHA-induced cytokine profile at two years of age and our in vitro findings support that specific bacterial species influence the T helper cell subsets. This suggests that dysbiosis in the early microbiota may modulate the risk of developing inflammatory conditions like allergy.     

A new in vitro system to study the effect of liquid phase turnover and pH on microbial fermentation of concentrate diets for ruminants

A simple and inexpensive in vitro feed evaluation system which estimates microbial fermentation by measuring gas production and allows for adjustment of liquid dilution rate has been design. Working conditions, in terms of substrate and inoculum proportions (experiment 1, EXP 1) and the liquid replacement schedule (experiment 2, EXP 2) were initially set. In EXP 1, three substrate concentrations (10, 20 and 30 mg/ml: S1, S2 and S3) and three inoculum proportions (0.10, 0.20 and 0.30; I10, I20 and I30) were assayed in a 3 × 3 factorial design, in three incubations of 24 h. Gas production (ml/g organic matter) with S1 was the highest, avoiding to be in excess for microbial colonisation and activity and inoculum I20 limited inoculum contribution by self-fermentation; they were hence considered for further trials. In EXP 2, two liquid replacement schedules (every hour from 0 to 12 h and every 3 h from 12 to 24 h, HF, and every 2 h from 0 to 12 h and every 4 h from 12 to 24 h, LF) were compared with a non-replacement (NR) treatment, using three bottles per treatment. No differences among treatments were detected, and thus the LF schedule was chosen because it was easier to carry out. Once these working conditions were adjusted in EXP 1 and 2, the effect of fluid turnover and incubation pH on the extent of microbial fermentation in a mixed diet (experiment 3, EXP 3) was studied in two incubations. Two liquid turnover rates (0.06 and 0.10 h⁻¹) were compared with a non-replacement treatment at two different pH (6.8 and 6.1) in a 3 × 2 factorial design. Liquid outflow increased gas production, dry matter disappearance (DMd) and total volatile fatty acid (VFA) concentration at both pH (P>0.01), but protozoal numbers were unaffected. Liquid replacement at pH 6.8 did not affect the VFA profile, but the acetate proportion diminished (P=0.04) and the propionate increased (P<0.001), with replacement at pH 6.1. Increasing the fluid turnover rate from 0.06 to 0.10 h⁻¹ reduced gas production from 8 to 20 h of incubation (P<0.05) but it did not alter the VFA profile. The volume of gas produced decreased (P<0.05) with incubation pH, as did DMd (P=0.04) and protozoal (P=0.03) and total VFA (P=0.002) concentration. The acetate proportion was lower (P<0.001) and that of propionate and butyrate was higher (P<0.001) at pH 6.1 than at pH 6.8. Both liquid turnover and pH must be taken into consideration for in vitro evaluation of mixed and concentrate diets for ruminants and this system provides a simple and inexpensive method for doing this.     

Neogene planktonic foraminiferal biostratigraphy and evolution: Equatorial to subantarctic, South Pacific

Detailed planktonic foraminiferal zonations have been established for the Neogene (Latest Oligocene through present) in six DSDP sites in the South Pacific ranging from equatorial to subantarctic latitudes (48°S). Two basic zonal schemes are readily recognized: tropical and temperate. The tropical zonation is best developed in DSDP Site 289 and the temperate zonation in Sites 206, 207A and 284. Tropical and temperate zonations can be linked by a warm subtropical scheme in Site 208, because this sequence includes a mixture of tropical and temperate elements. A site located close to the Subantarctic Convergence (Site 281) contains a zonation largely of temperate character, but the present of cooler elements and some differences in biostratigraphic ranges have required a slightly different biostratigraphic scheme.Although two broad schemes are recognized, none of the biostratigraphic sequences are identical between any of the sites. This reflects differences in biogeography, evolution and diachronous extinction at various latitudes during the entire Neogene. Diachronism in biostratigraphic ranges continue to create difficulties in correlation across such wide latitudes.Our detailed work has required the establishment of new biostratigraphic zonations in certain parts of the Neogene sequence and modifications in some other parts. Otherwise, previously established schemes are followed as closely as possible. In the temperate region, a new zonation has been established for the Early Miocene to early Middle Miocene. For the remainder of the Neogene the zonation of Kennett (1973) has been largely used. The tropical zonation of Blow (1969) is employed in the equatorial Site 289, but with further subdivisions for Zones N4 and N17. For areas intermediate between tropical and temperate latitudes (Site 208), a modified Early Miocene zonation is established based on changes in tropical and temperate elements.The zonal schemes are established on taxa that exhibit both diachronous and isochronous ranges across the latitudes. Zones that are at least partly diachronous include the Globigerinoides trilobus and Globorotalia miozea Zones of Early Miocene age; perhaps the Globorotalia mayeri Zone (its base) of the Middle Miocene; the Globorotalia conomiozea Zone of the Late Miocene; and the Globorotalia crassaformis Zone of the Early Pliocene.A large number of datum levels are recognized based on first evolutionary appearances or extinctions. The most widely applicable datums are as follows: latest Oligocene — Globigerinoides F.A.; Early Miocene — Globoquadrina dehiscens, F.A., Globorotalia kugleri L.A., Catapsydrax dissimilis L.A. and Praeorbulina glomerosa F.A.; Middle Miocene — Orbulina suturalis F.A., Globorotalia peripheroacuta F.A., Fohsella lineage L.A., Globorotalia mayeri L.A.; Late Miocene — “Neogloboquadrina” continuosa L.A., Globoquadrina dehiscens L.A., Globorotalia cibaoensis F.A.; Early Pliocene — Globorotalia puncticulata F.A., Globorotalia margaritae F.A.; Early Pleistocene — Globorotalia truncatulinoides F.A. A number of other datums are identified which assist with correlation over more restricted latitudinal ranges.The evolution of most Neogene planktonic foraminifera is now well established for a wide range of water masses. Evolutionary lineages are primarily centered in the temperate and tropical regions. Tropical lineages have recently been reviewed by Srinivasan and Kennett (1981) and are not discussed in detail here. However, Sphaeroidinellopsis seminulina is now considered to have evolved directly into S. paenedehiscens during the Late Miocene and S. subdehiscens Blow is considered to be junior synonym of S. seminulina.A new evolutionary lineage is recognized in the warm subtropics (Site 208) whereby Globigerina woodi woodi gave rise to Globigerinoides subquadratus via Globigerina brazieri. The discovery of this lineage clearly demonstrates that Globigerinoides is a polyphyletic “genus”. Another major phylogenetic lineage is recognized within the temperate globorotaliids of Early Miocene age as follows: “N.” continuosa → Globorotalia zealandica incognita → G. zealandica zelandica → G. praescitula → G. miozea. Although parts of this lineage have been recognized earlier, the entire phylogeny has previously been underscribed.A new Early to Middle Miocene lineage is recognized in the subantarctic to temperate areas which involve a transition from Globorotalia praescitula to G. challengeri n. sp. via intermediate forms.Two major Neogene globorotaliid lineages — the Menardella of the tropics and Middle Miocene to Recent forms of Globoconella of the temperate areas — are both considered to have evolved from Globorotalia praescitula beginning in the Early Miocene. This evolution initially was restricted to temperate areas but has since separated into distinctly tropical and temperate phylogenetic elements.     

Ensiling and subsequent ruminal degradation characteristics of barley tea grounds treated with contrasting additives

The objective of this experiment was to determine the ensiling characteristics and in situ degradation of barley tea grounds (BTG) treated with contrasting additives. The BTG was ensiled without additive (control), or with mixtures of lactic acid bacteria and Acremonium cellulase (LAB + AUS), formic acid (FA), and sodium hydroxide (NaOH) for a period of 60 days. Three ruminal-fistulated steers were used to determine in situ degradabilities of dry matter (DM), crude protein (CP) in the BTG and its silages. In the LAB + AUS treated silage, the pH value (P<0.001) and ammonia-N content (P=0.007) were lower and the lactic acid content (P<0.001) was higher than the control. The FA and NaOH treated silages were well preserved as indicated by low ammonia-N content (P=0.007), no propionic acid and butyric acid. The LAB + AUS treated silage had higher rapidly degradable fraction of DM and CP than the control and BTG (P<0.001). The NaOH treated silage had higher contents of rapidly degradable fraction, slowly degradable fraction and effective degradability of DM and CP than control and BTG (P<0.01).     

Effects of Brosimum alicastrum and Lysiloma latisiliquum mixtures on voluntary intake, nutrient digestibility and nitrogen balance in sheep fed tropical pastures

Effects of supplementing tree foliage mixtures on voluntary intake, apparent digestibility and N balance was evaluated using Pelibuey sheep fed low quality diets. Five treatments were examined in a 5 × 5 Latin square design, which consisted of a basal diet of grass (Sorghum halepense) hay supplemented with Brosimum alicastrum (B) and Lysiloma latisiliquum (L) at the following rates (g DM/kg diet): B264, L0; B198, L66; B132, L132; B66, L198 and B0, L264. Additionally, an in situ degradability evaluation was completed with two ruminally cannulated cows. Neutral detergent fibre (NDF), acid detergent insoluble N (ADIN), lignin(sa) and total phenols (TP) were higher (P<0.01) in L. latisiliquum versus B. alicastrum. Daily intake (g/kg LW^0.75/day) of DM (from 98 to 73) and OM (from 88 to 66) decreased quadratically (P<0.01), whereas CP (from 8.0 to 5.6) and ME (from 7.7 to 5.2, MJ/sheep/day) reduced linearly (P<0.01), as L. latisiliquum increased in the diet. Apparent digestibility of DM (from 0.486 to 0.445), OM (from 0.511 to 0.458) and CP (from 0.417 to 0.198) decreased linearly (P<0.01) and was associated with a low ruminal in situ CP degradability of L. latisiliquum. The decrease in N intake and digestibility induced lower (P<0.01) N retention (from 2.7 to 0.1 g/sheep/day). Although the incremental substitution of B. alicastrum with L. latisiliquum negatively affected intake, rumen degradation, digestibility and N balance, results indicate that this foliage mixture, but with no more than 132 g DM/kg diet of L. latisiliquum, could be used as a supplementation strategy to sheep fed low quality forage without negative effects on voluntary intake.     

Characterization of protein fractions and amino acids in ensiled alfalfa treated with different chemical additives

Formic acid, formaldehyde, tannic acid or mixtures of two were studied on their effects on ensiled alfalfa (Medicago sativa L.) amino acids and N fractions by the Cornell Net Carbohydrate and Protein System (CNCPS). The alfalfa forage was a second cut and was wilted to a mean over-dry dry matter (DM) content of 330 g/kg. All silages were prepared as mini-silos using 100 ml polypropylene centrifuge tubes (50 g) on a small laboratory-scale, with the additives added in 20 ml aliquots/kg herbage fresh weight (FW). After 35 d of ensiling, most of forage true protein was converted to fraction A and all of the added additives reduced fraction A content in the ensiled forages (P<0.05). The content of fraction B₁ in all of the additive-treated silages was higher (P<0.05) than that in control silage. Large proportions of true protein in the tannic acid/formaldehyde- and formic acid/formaldehyde-treated silages were fractions B₂ and B₃, respectively. No difference was observed on fraction C content between the control silage and silages treated with additives except for the formaldehyde or tannic acid-treated silages. Amino acids were well preserved in additive-treated silages compared with the control silage. Concentration of total amino acid was higher in formic acid-treated silages than that in the control and the other additive-treated silages (P<0.05). The pattern of changes in individual amino acid in all of the silages indicated that branched chain amino acids and methionine were relatively well preserved during fermentation but the basic and acidic amino acids were not.     

Temperature adaptation and the contractile properties of live muscle fibres from teleost fish

Summary The contractile properties of swimming muscles have been investigated in marine teleosts from Antarctic (Trematomus lepidorhinus, Pseudochaenichthys georgianus), temperate (Pollachius virens, Limanda limanda, Agonis cataphractus, Callionymus lyra), and tropical (Abudefduf abdominalis, Thalassoma duperreyi) latitudes. Small bundles of fast twitch fibres were isolated from anterior myotomes and/or the pectoral fin adductor profundis muscle (m. add. p). Live fibre preparations were viable for several days at in vivo temperatures, but became progressively inexcitable at higher or lower temperatures. The stimulation frequency required to produce fused isometric tetani increased from 50 Hz in Antarctic species at 0°C to around 400 Hz in tropical species at 25°C. Maximum isometric tension (P_o) was produced at the normal body temperature (NBT) of each species (Antarctic, 0–2°C; North Sea and Atlantic, 8–10°C; Indo-West Pacific, 23–25°C). P₀ values at physiological temperatures (200–300 kN·m^–2) were similar for Antarctic, temperate, and tropical species. A temperature induced tension hysteresis was observed in muscle fibres from some species. Exposure to <0°C in Antarctic and <2°C in temperate fish resulted in the temporary depression of tension over the whole experimental range, an effect reversed by incubation at higher temperatures. At normal body temperatures the half-times for activation and relaxation of twitch and tetanic tension increased in the order Antarctic>temperate>tropical species. Relaxation was generally much slower at temperatures <10°C in fibres from tropical than temperate fish. Q₁₀ values for these parameters at NBTs were 1.3 2.1 for tropical species, 1.7–2.6 for temperate species, and 1.6–3.5 for Antarctic species. The forcevelocity (P-V) relationship was studied in selected species using iso-velocity releases and the data below 0.8 P₀ iteratively fitted to Hill's equation. The P-V relation at NBT was found to be significantly less curved in Antarctic than temperate species. The unloaded contraction velocity (V_max) of fibres was positively correlated with NBT increasing from about 1 muscle fibre length·s^–;1 in an Antarctic fish (Trematomus lepidorhinus) at 1°C to around 16 muscle fibre lengths·s^–1 in a tropical species (Thalassoma duperreyi) at 24°C. It is concluded that although muscle contraction in Antarctic fish shows adaptations for low temperature function, the degree of compensation achieved in shortening speed and twitch kinetics is relatively modest.Abbreviations ET environmental temperature - m. add. p major adductor profundis - m. add. s. major adductor superficialis - NBT normal body temperature - P ₀ maximum isometric tension - P-V force velocity - SR sarcoplasmic reticulum - T 1/2 a half activation time - T 1/2 r half relaxation time - V _max unloaded contraction     

Methane emission by goats consuming diets with different levels of condensed tannins from lespedeza

Twenty-four yearling Boer × Spanish wethers (7/8 Boer; initial body weight (BW) of 34.1 ± 1.02 kg) were used to determine effects on methane (CH₄) emission of dietary levels of a condensed tannin (CT)-containing forage, Kobe lespedeza (Lespedeza striata; K), and a forage very low in CT, sorghum-sudangrass (Sorghum bicolor; G). Treatments were dietary K levels (dry matter (DM) basis) of 1.00, 0.67, 0.33, and 0 (100, 67, 33, and 0 K, respectively). Forages were harvested daily and fed at approximately 1.3 times maintenance metabolizable energy requirement. The experiment lasted 21 days, with most measures on the last 8 days. The CT concentration was 0.3 and 151 g/kg DM in G and K, respectively. DM intake was similar among treatments (i.e., 682, 675, 654, and 648 g/day; S.E. = 30.0) and gross energy (GE) digestibility increased linearly (P<0.05) with decreasing K (0.472, 0.522, 0.606, and 0.666 for 100, 67, 33, and 0 K, respectively). CH₄ emission changed quadratically (P<0.05) with decreasing K (10.9, 13.8, 17.6, and 26.2 l/day; 32, 42, 57, and 88 kJ/MJ GE; 69, 81, 94, and 133 kJ/MJ digestible energy for 100, 67, 33, and 0 K, respectively). In vitro CH₄ emission by incubation of ruminal fluid for 3 weeks with a medium for methanogenic bacteria and other conditions promoting activity by methanogens also was affected quadratically (P<0.05) by K level (7.0, 8.1, 9.2, and 16.1 ml for 100, 67, 33, and 0 K, respectively). The total bacterial count of ruminal samples was similar among K levels, but the number of total protozoa increased linearly (P<0.05) as K declined (8.3, 11.8, 15.6, and 27.1 × 10⁵ ml⁻¹ for 100, 67, 33, and 0 K, respectively). The CT-containing forage K decreased CH₄ emission by goats regardless of its feeding level, although the effect per unit of K increased with decreasing K. Forage type (i.e., legume versus grass) may have contributed to the effect of K on CH₄ emission, but most of the change appeared attributable to CT, which appeared to directly impact activity of methanogenic bacteria, although alterations of protozoal activity could have been involved. These findings suggest that relatively low dietary levels of CT could be employed to lessen CH₄ emission without a marked detrimental effect on other conditions such as total tract protein digestion.     

CYLINDROSPERMOPSIS RACIBORSKII (CYANOBACTERIA) INVASION AT MID‐LATITUDES: SELECTION,WIDE PHYSIOLOGICAL TOLERANCE,ORGLOBALWARMING?1

The tropical bloom‐forming cyanobacterium Cylindrospermopsis raciborskii (Woloszynska) Seenaya et Subba Raju is causing increasing concern because of its potential toxicity and invasive behavior at mid‐latitudes. This species has recently been identified in several temperate areas and was first reported in France in 1994, but the mechanisms underlying this acclimation process remain to be elucidated. We performed a range of laboratory experiments in an attempt to identify the physiological characteristics that could account for this behavior. We investigated the three following hypotheses: 1) clones of C. raciborskii adapted to temperate climate have been selected as it advanced north, 2) C. raciborskii has high physiological tolerance that allows it to prosper in a wide range of conditions, and 3) changes inclimate (global warming) have favored the development of C. raciborskii in mid‐latitudes. Ten strains of C. raciborskii from Australia n=1), Brazil (2), France (2), Germany (1), Hungary (1), Mexico (1), and Senegal (2) were cultured at different temperatures and light intensities. The in vitro growth parameters (μ and T_opt) were the same for the tropical and temperate strains. All 10 strains displayed positive net growth in a wide range of temperatures (from 20 to 35°C) and light intensities (from 30 to 400 μmol photons·m^?2·s^?1), with maximum growth rates at around 30°C and 80 μmol photons·m^?2·s^?1. This suggests that the colonization of mid‐latitudes by C. raciborskii may result from a combination of its ability to tolerate a rather wide range of climatic conditions and the global warming phenomenon, which provides this species with better environmental conditions for its growth.     

<Emphasis Type="Italic">In vitro</Emphasis> collecting (IVC). I. The effect of collecting method and antimicrobial agents on contamination in temperate and tropical collections

Summary In vitro collecting is the process of initiating tissue cultures in the field. In order for in vitro collecting to be broadly available as a technique for collecting plant germplasm, the levels of contamination in such cultures must be controlled. Two techniques for in vitro collecting were compared: leaf punch and needle collecting. The effectiveness of these methods for collecting leaf and stem tissues from plants at tropical and temperature sites was compared. Stem tissue collected by the needle collecting method gave cultures with an average contamination percentage of 31% and 16%, from the tropical and temperate sites, respectively, while with the leaf punch method, average contamination percentages were 90% and 69%. The effectiveness of antimicrobial agents in reducing contamination in leaf punch cultures was evaluated. Addition of the fungicide benlate and the antibiotics cefotaxime and vancomycin, to the leaf punch collections reduced contamination to an average of 30% in the tropical collections and 35% in the temperate collections. Over 90% of both tropical and temperature species collected in multiple samples of 10 or more had at least one clean sample using this medium. The use of either the leaf punch method in combination with a fungicide and antibiotics or the needle collecting technique yielded a high percentage of clean tissues for study and growth.     

A comparison of the responses of tropical and temperate flies (Diptera: Sarcophagidae) to cold and heat stress

Summary Two flesh fly species from the tropical lowlands (Peckia abnormis and Sarcodexia sternodontis) were more susceptible to both cold-shock and heatshock injury than temperate flies (Sarcophaga crassipalpis and S. bullata) and a fly from a tropical high altitude (Blaesoxipha plinthopyga). A brief (2-h) exposure to 0°C elicits a protective response against subsequent cold injury at–10°C in the temperate flies and in B. plinthopyga but no such response was found in the flies from the tropical lowlands. However, both tropical and temperate flies could be protected against heat injury (45°C) by first exposing them to a mild heat shock (2 h at 40°C). The supercooling point is not a good indicator of cold tolerance: supercooling points of pupae were similar in all species, ranging from–18.9 to–23.0°C, and no differences were found between the tropical and temperate species. Among the temperate species, glycerol, the major cryoprotectant, can be elevated by short-term exposure to 0°C, but glycerol could not be detected in the tropical flies. Low-temperature (0°C) exposure also increased hemolymph osmolality of the temperate species, but no such increase was observed in the tropical lowland species. Adaptations to temperature stress thus differ in tropical and temperate flesh flies: while flies from both geographic areas share a mechanism for rapidly increasing heat tolerance, only the temperate flies appear capable of responding rapidly to cold stress. The presence of a heat shock response in species that lack the ability to rapidly respond to cold stress indicates that the biochemical and physiological bases for these two responses are likely to differ.