Escape protein supplementation of growing steers grazing stargrass

A grazing trial utilizing 35 individually supplemented growing steers (211±42 kg initial body weight (BW)) was conducted to study the effect of supplemental escape protein on the performance of steers grazing on stargrass (Cynodon plectostachyus) during the dry season. N in supplements was 100%, 50%, or 0% natural protein (bloodmeal, coconut meal, and soybean meal), and 0%, 50% or 100% urea. All steers received 2 kg of supplement dry matter (DM) (2.2% N) daily during the 90 days of the experiment. Steers fed the urea supplement had the lowest ADG (0.97 kg day⁻¹). There was a linear (P<0.05) response in ADG to the natural protein level (50 and 100%) in supplements containing bloodmeal (1.11 and 1.21 kg day⁻¹) and coconut meal (1.05 and 1.21 kg day⁻¹), but no response was observed with soybean meal (1.01 and 1.0 kg day⁻¹). Forage intake was not affected by supplementation. As a result of the growth response observed for supplements containing bloodmeal and coconut meal above the urea-based and soybean meal supplements, it was concluded that growing ruminants grazing stargrass in the dry season were deficient in escape protein. ©1997 Elsevier Science B.V.     

The influence of dry period feeding on liver fat and postpartum performance of dairy cows

Animal performances were monitored in 30 Friesean dry cows (18 multiparous, MP-cows and 12 primiparous, PP-cows) starting six weeks before calving to eight weeks after calving. The cows were kept indoors and fed individually with a prepartum diet containing either low, moderate or high energy (0.75, 1.00 or 1.25 of the calculated ME requirement) and supplemented with low (0.3 kg day⁻¹) or high (1.5 kg day⁻¹) rapeseed meal (RSM). The diets were consisted of 1.5 kg hay, 20–25 kg wilted grass silage and grain (barley and oat, 1 : 1 DM basis) with RSM. The average ME intake during six weeks of prepartum was 75, 97 and 123 MJ day⁻¹ on three different energy levels. After calving the cows were fed grass silage ad lib, 6.5 kg (primiparous) or 8.5 kg (multiparous) grain and 1.5 kg RSM. Liver biopsy and blood samples were taken to determine differences in metabolites due to diet and parity. Reduction of prepartum energy allowance to 0.75 of the moderate level did not affect the calf's birth weight and colostrum composition. Cows fed prepartum low energy–high rapeseed meal diet (LEHR) showed a faster increase in feed intake and milk production after parturition. Continuous low feed intake and milk production were observed in cows fed a prepartum high energy–low rapeseed meal diet (HELR). Milk yield was constantly lower for cows fed prepartum high energy diet, the difference being significant (p<0.01 and p<0.05) at six and seven weeks of lactation. Overall milk yield also showed a linear decrease with energy feeding level and an increase (p<0.10) with RSM feeding levels. Prepartum high energy feeding increased (p<0.01) milk protein content. Neither energy nor RSM feeding level affected the liver fat infiltration of the experimental cows. Cow parity however showed a significant influence on liver fat content, β-hydroxybutyrate (BHBA) and plasma glucose. A week before calving, the plasma concentration of essential amino acids (EAA) was lower for cows fed prepartum low energy diet. During the first four weeks of lactation, the concentrations of plasma non-esterified fatty acids (NEFA) and insulin were similar for all the treatment group.     

Growth and loss of distal tissue in blades of Lessonia nigrescens and Lessonia trabeculata (Laminariales)

Meristematic growth and loss of distal tissue from blades of two ecologically important species in the south-east Pacific, Lessonia nigrescens and Lessonia trabeculata, was evaluated during 1 year. Comparative growth was determined by a hole-punch method, loss of distal tissue from the blades was determined by subtracting final blade length (with loss) from expected blade lengths (without loss); growth and tissue loss were transformed to fresh biomass units for calculation of inter-algae differences. The results showed that blade elongation rate increased at the beginning of spring, and declined towards the end of summer, with mean values between 0.40 and 0.08 cm day⁻¹ for L. nigrescens, and 0.65–0.17 cm day⁻¹ for L. trabeculata. Loss of distal tissue varied seasonally when examined as length units for both species; with mean values between 0.24 and 0.10 cm day⁻¹ for L. nigrescens, and 0.51–0.25 cm day⁻¹ for L. trabeculata. Variations in fresh biomass units were only observed in Lessonia trabeculata, increasing in spring, with mean values to 0.13 g (fresh weight) day⁻¹. Annual growth and loss of distal tissue were higher in L. trabeculata (0.41 and 0.39 cm day⁻¹, respectively) than in L nigrescens (0.19 and 0.15 cm day⁻¹). When growth and tissue loss were considered as fresh biomass, monthly gains significantly outweighed loss of distal tissue in both species, but parallel results based on length data followed a different trend. L. trabeculata released about 50% of its growth biomass as particulate organic matter, while the comparative value for L. nigrescens was about 20%.     

Growth performances and carcass traits of Ovin Martinik lambs fed various ratios of tropical forage to concentrate under intensive conditions

Studies were conducted to determine the effects of supplementation upon intake, growth and carcass traits of Ovin Martinik hair sheep. Forty lambs weighing 20 ± 3.7 kg live weight (LW) were reared after weaning in individual pens during a 4-month experimental study. Four supplement levels were compared (10 lambs per treatment): the L0 group received the basal diet (tropical forage 0.75 UFL and 73 PDIN g kg⁻¹ DM, INRA system) without concentrate, the L150, L300 and L600 groups were offered in addition 150, 300 and 600 g day⁻¹ of concentrate (1.15 UFL and 151 PDIN g kg⁻¹ DM), respectively. Intake (5 days a week) and growth patterns (every fortnight) were measured on the 40 lambs, while digestibility measurements (five times throughout the experimental period) were determined on half of the animals fitted with faecal bags and which were slaughtered according to the standard procedure at 33–35 kg live weight for assessment of carcass traits and meat quality.The total dry matter intake (DMI) and digestibility increased (P < 0.01) with the inclusion ratio of the concentrate in the diet: 82 g DM kg W^−0.75 vs. 97 g DM kg W^−0.75 and 65% vs. 73% between the two extreme groups, whereas forage DMI decreased and the forage substitution ratio was calculated to be 0.65. Growth and feed/gain ratios significantly (P < 0.01) improved among the groups L0, L150 and L300, from an average of 134–188 g day⁻¹ and 7.0–5.7, respectively. The differences observed between L300 and L600 were not significant. Carcass weight and dressing out percentages significantly (P < 0.01) increased with the addition of concentrate to the ration: from 13.1 to 15.6 kg and from 39 to 46%, from L0 to L600, respectively. Consequently, the weights of the different cuts of the carcass varied. There was a significant effect of the supplementation level on the accumulation of internal fat tissues: the kidney fat weight increased from 107 to 237 g from the L0 to the L600 groups. No significant effect was observed on the carcass quality scores, the ultimate pH and the colour parameters of the meat.By increasing the nutritional density of the diet, it was possible to obtain well-conformed and heavier carcasses, with no apparent detrimental effect on the quality. The optimal supplement supply with good quality grass would be 300 g day⁻¹ in our conditions.     

Effect of nutrient pulses on photosynthesis of Chaetomorpha linum from a shallow Mediterranean coastal lagoon

The influence of nitrogen and phosphorus pulses on Chaetomorpha linum (Muller) Kutzing growth and photosynthesis was studied in laboratory experiments. Photosynthesis and growth of C. linum from Tancada lagoon seems limited by both nitrogen and phosphorus, as indicated by the high rate (4.7–11.6 mg O₂ g⁻¹ dry weight h⁻¹) of light-saturated photosynthesis (P_m) and growth rates observed under nitrogen plus phosphorus enrichment in relation to enrichment by nitrogen alone (2.9–7.6 mg O₂ g⁻¹ dry weight h⁻¹). Significant increase in nitrogen and phosphorus content as percentage of dry weight was observed in C. linum fertilized with a single nutrient or with nitrogen plus phosphorus. In Tancada lagoon, when availability of nitrogen to primary producers is by pulses, an increase of nitrate concentration in the water column (from 6 to 100 μM) has a greater effect on growth of C. linum (growth rate: 0.13 day⁻¹) than an increase in ammonium concentration (from 20 to 100 μM and growth rate: 0.11 day⁻¹). For a given thallus nitrogen content (0.6–1.4% N), both P_m and the photosynthetic efficiency (α) normalized to dry weight were correlated (r² = 0.73, p < 0.005) indicating that variations in electron transport were coupled to variations in C-fixation capacity. Optimizing both α and P_m may be a general characteristic of thin-structured opportunistic algae in more variable estuarine environments.     

Growth of three submerged plants below different densities of Nymphoides peltata (S. G. Gmel.) Kuntze

We evaluated one-sided competition from the floating-leaved plant Nymphoides peltata (non-indigenous in Sweden) on three submerged plant species, Ceratophyllum demersum, Elodea canadensis and Ranunculus circinatus, in a controlled experiment. The three submerged species were allowed to grow for 21 days in the absence of N. peltata and with the species present at densities of approximately 33, 66 and 100% cover. All species retained a positive relative growth rate (RGR) based on length at all N. peltata densities, but responded with negative growth based on weight for several treatments. C. demersum achieved RGR of 0.03 day⁻¹ in the absence of N. peltata, RGR of 0.02 day⁻¹ in the lowest N. peltata density but negative RGR in the two denser treatments. E. canadensis responded similarly with RGR of 0.04 day⁻¹ in the absence of N. peltata, RGR of 0.01 day⁻¹ in the lowest N. peltata density and negative RGR in the two denser treatments. R. circinatus, on the other hand, never achieved positive RGR based on weight. These results suggest that one-sided competition from floating-leaved plants has a profound effect on the submerged plant community.     

Interaction of dietary zinc and growth implants on weight gain,carcass traits and zinc in tissues of growing beef steers and heifers

In two separate studies, 60 beef heifers (379 kg BW) and 60 beef steers (348 kg BW) were randomly assigned to six treatments in 2×3 factorial arrangements. The treatments were with or without Synovex^® implants combined with either a control diet or diets supplemented with 200 ppm Zn from ZnSO₄ or zinc methionine (Zn-Met). Near the mid-point of the feeding periods, cattle were vaccinated with a modified live virus and subsequent titers and concentrations of immunoglobulin G (IgG) were measured. Liver and blood samples were obtained 1 week prior to the start of the experiments and at intervals during the experiments. In experiment 1, average daily gains of beef heifers were (P<0.05) affected by the interaction of implant and source of dietary Zn. Compared to control and ZnSO₄ treatments, supplementation with Zn-Met increased (P<0.05) the concentration of Zn in serum. Antibody titers and concentrations of IgG in serum were highest (P<0.05) in heifers fed ZnSO₄ compared to heifers fed the control or Zn-Met supplemented diets. The Synovex-H^® implant reduced the concentrations of Zn and Cu in liver. In experiment 2, Synovex-S^® implants improved (P<0.05) weight gains of steers supplemented with 200 ppm dietary Zn from ZnSO₄ compared to non-implanted steers. However, the implant had no effect when Zn-Met was the dietary Zn source. The implant increased (P<0.05) concentrations of Zn in liver of steers supplemented with 200 ppm dietary Zn and reduced Zn in liver of steers fed the control diet. Implanted steers had higher (P<0.05) Cu status and IgG concentrations in serum than non-implanted steers. Steers supplemented with either ZnSO₄ or Zn-Met had greater (P<0.05) concentrations of Zn in liver and plasma than steers fed the control diet. These results indicate both the level and source of Zn supplementation in diets of feedlot cattle affect their response to growth implants.     

Chara hispida beds as a sink of nitrogen: Evidence from growth,nitrogen uptake and decomposition

Chara hispida forms dense beds (0.78–0.95 kg DW m⁻²) in Colgada Lake. The ability of Chara meadows to act as a nitrogen source or sink was evaluated by the following methods: (1) investigating Chara growth, (2) nitrogen incorporation and decomposition laboratory experiments and (3) relating experimental results to field conditions. Sediment oospores were germinated in large aquaria and observed growth rates were 0.001 m day⁻¹ (shoot length) and 0.0002 g day⁻¹ (dry weight). Nitrogen uptake rates were determined by addition of K¹⁵NO₃ during two different periods of Chara growth and the rates were 1.21 and 3.86 μM g DW⁻¹ h⁻¹ when charophytes were 166 days old (not sexually mature) and 323 days old (sexually mature), respectively. After the uptake experiments, the same charophytes were allowed to decompose within two types of litter bags (3 mm-pore litter bags and entire, non-porous plastic litter bags). Decomposition rates of Ch. hispida were 0.016 and 0.009 day⁻¹ in perforated and non-perforated bags, respectively, and fit a negative exponential model. The nitrogen release rate, calculated as the disappearance of N content from Chara tissues, was 0.012 day⁻¹ and there were no statistically significant differences between the values from the two different bag types. The dissolved organic nitrogen concentrations in aquarium and non-perforated litter bags waters increased linearly with time due to the leaching of soluble compounds from Chara. The rate of N loss from Chara tissues, total nitrogen and dissolved organic nitrogen release rates and the decrease in initial dry weight rate were all lower than the daily rate of Chara N uptake. By extrapolating laboratory data to field situations, we determined that approximately 38% of the N taken up by charophytes in Colgada Lake during the growth period is retained. Given the high charophyte biomass in the lake, its ability to incorporate nitrogen, its low decomposition rate and its ability to over-winter, we conclude that Chara beds could be acting as nitrogen sinks in this ecosystem.     

Growth of the fungus Paecilomyces lilacinus with n-hexadecane in submerged and solid-state cultures and recovery of hydrophobin proteins

The filamentous fungus Paecilomyces lilacinus was grown on n-hexadecane in submerged (SmC) and solid-state (SSC) cultures. The maximum CO₂ production rate in SmC (V_max = 11.7 mg CO₂ L_g⁻¹ day⁻¹) was three times lower than in SSC (V_max = 40.4 mg CO₂ L_g⁻¹ day⁻¹). The P. lilacinus hydrophobin (PLHYD) yield from the SSC was 1.3 mg PLHYD g protein⁻¹, but in SmC, this protein was not detected. The PLHYD showed a critical micelle concentration of 0.45 mg mL⁻¹. In addition, the PLHYD modified the hydrophobicity of Teflon from 130.1 ± 2° to 47 ± 2°, forming porous structures with some filaments <1 μm and globular aggregates <0.25 μm diameter. The interfacial studies of this PLHYD could be the basis for the use of the protein to modify surfaces and to stabilize compounds in emulsions.     

Microbial protein synthesis in growing-finishing bulls estimated from the urinary excretion of purine derivatives

In the frame of a feeding experiment with three periods, balance trials were carried out with 18 double-muscled Belgian White–blue bulls, allocated to one of four feeding regimes. The ration consisted of maize silage and concentrate in the ratio of 35:65 on DM basis. Six concentrates were formulated to supply three levels of protein and energy. The three periods corresponded to distinct live weight intervals of 360–460, 460–570 and 570–680 kg. In the first feeding regime, low protein and intermediate energy level were given during the whole trial; in the second regime, protein level decreased at a constant intermediate energy level; in the third regime, the energy level increased at a constant high protein level; in the fourth regime, protein level decreased simultaneously with an increase in the energy level. Total daily intake varied from 6.2 to 9.7 kg for dry matter (DM), from 65 to 106 MJ for metabolisable energy (ME) and from 719 to 1326 g for crude protein (CP) (50 animal observations). At the end of each period, the excretion of the purine derivatives (PD), allantoin and uric acid, was measured after total urine collection during 4 days to estimate microbial nitrogen supply to the duodenum (MN_PD). The effect of the intake of DM, organic matter (OM), digestible OM (DOM), digestible carbohydrates (DCHO), total digestible nutrients (TDN), rumen fermentable OM (FOM), metabolisable energy (ME), fermentable ME (FME), CP, digestible CP (DCP) and rumen degradable protein (RDP) on PD and MN_PD was examined. Further, the relationship of MN_PD to MN, calculated according to different systems, was examined. The amount (mean±SD) of allantoin excreted in urine was 147±23 mmol day⁻¹ and of uric acid 11±3 mmol day⁻¹. The MN_PD amounted to 97±21 g day⁻¹, varying from 57 to 154 g day⁻¹, and significantly increased with all measures of nutrient intake. The correlation coefficients, ranging from 0.45 for DCP to 0.57 for DOM and FME, were, however, not significantly different. MN_PD showed a larger variation and was on average lower than the potential MN values calculated from the intake of FOM (108±13 g day⁻¹), DCHO (141±16 g day⁻¹) and FME (109±13 g day⁻¹), similar to that calculated from the intake of RDP (99±14 g day⁻¹) and higher than the MN value calculated from the intake of TDN (84±11 g day⁻¹). The correlations of MN_PD to the calculated MN values ranged from 0.47 for MN_FOM to 0.59 for MN_FME, but were not significantly different.     

Germination fluctuation of toxic Alexandrium fundyense and A. pacificum cysts and the relationship with bloom occurrences in Kesennuma Bay,Japan

While cyst germination may be an important factor for the initiation of harmful/toxic blooms, assessments of the fluctuation in phytoplankton cyst germination, from bottom sediments to water columns, are rare in situ due to lack of technology that can detect germinated cells in natural bottom sediments. This study introduces a simple mesocosm method, modeled after previous in situ methods, to measure the germination of plankton resting stage cells. Using this method, seasonal changes in germination fluxes of toxic dinoflagellates resting cysts, specifically Alexandrium fundyense (A. tamarense species complex Group I) and A. pacificum (A. tamarense species complex Group IV), were investigated at a fixed station in Kesennuma Bay, northeast Japan, from April 2014 to April 2015. This investigation was conducted in addition to the typical samplings of seawater and bottom sediments to detect the dinoflagellates vegetative cells and resting cysts. Bloom occurrences of A. fundyense were observed June 2014 and February 2015 with maximum cell densities reaching 3.6 × 10⁶ cells m⁻² and 1.4 × 10⁷ cells m⁻², respectively. The maximum germination fluxes of A. fundyense cysts occurred in April 2014 and December 2014 and were 9.3 × 10³ cells m⁻² day⁻¹ and 1.4 × 10⁴ cells m⁻² day⁻¹, respectively. For A. pacificum, the highest cell density was 7.3 × 10⁷ cells m⁻² during the month of August, and the maximum germination fluxes occurred in July and August, reaching 5.8 × 10² cells m⁻² day⁻¹. Thus, this study revealed the seasonal dynamics of A. fundyense and A. pacificum cyst germination and their bloom occurrences in the water column. Blooms occurred one to two months after peak germination, which strongly suggests that both the formation of the initial population by cyst germination and its continuous growth in the water column most likely contributed to toxic bloom occurrences of A. fundyense and A. pacificum in the bay.     

Characteristics of simultaneous organic and nutrient removal in a pilot-scale vertical submerged membrane bioreactor (VSMBR) treating municipal wastewater at various temperatures

A pilot-scale vertical submerged membrane bioreactor (VSMBR) with anoxic and oxic zones in one reactor was operated in an attempt to reduce the problems concerning effective removal of organic matter and nutrients from municipal wastewater. Source water with total chemical oxygen demand (TCOD)/total nitrogen (TN) ratio of 5.5 was treated at various temperatures (13–25 °C) over an interval of about 1 year. As a result, total suspended solid (TSS) and TCOD were removed by 100% and higher than 98%, respectively. Moreover, the average removal efficiencies of TN and total phosphorus (TP) were found to be 74% and 78% at 8 h-hydraulic retention time (HRT) and 60-days sludge retention time (SRT). Under these conditions, the specific removal rates (SRR) of TN and TP were found to be 0.093 kg N m⁻³ day⁻¹ and 0.008 kg P m⁻³ day⁻¹, and the daily production of excess sludge (DPES), 0.058 kg TSS day⁻¹.     

Salt effects on β-glucosidase: pH-profile narrowing

Salts inhibit the activity of sweet almond β-glucosidase. For cations (Cl⁻ salts) the effectiveness follows the series: Cu⁺², Fe⁺² > Zn⁺² > Li⁺ > Ca⁺² > Mg⁺² > Cs⁺ > NH₄⁺ > Rb⁺ > K⁺ > Na⁺ and for anions (Na⁺ salts) the series is: I⁻ > ClO₄⁻ > ⁻SCN > Br⁻ ≈ NO₃⁻ > Cl⁻ ≈ ⁻OAc > F⁻ ≈ SO₄^− 2. The activity of the enzyme, like that of most glycohydrolases, depends on a deprotonated carboxylate (nucleophile) and a protonated carboxylic acid for optimal activity. The resulting pH-profile of k_cat/K_m for the β-glucosidase-catalyzed hydrolysis of p-nitrophenyl glucoside is characterized by a width at half height that is strongly sensitive to the nature and concentration of the salt. Most of the inhibition is due to a shift in the enzymic pK_as and not to an effect on the pH-independent second-order rate constant, (k_cat/K_m)^lim. For example, as the NaCl concentration is increased from 0.01 M to 1.0 M the apparent pK_a1 increases (from 3.7 to 4.9) and the apparent pK_a2 decreases (from 7.2 to 5.9). With p-nitrophenyl glucoside, the value of the pH-independent (k_cat/K_m)^lim (= 9 × 10⁴ M^− 1 s^− 1) is reduced by less than 4% as the NaCl concentration is increased. There is a similar shift in the pK_as when the LiCl concentration is increased to 1.0 M. The results of these salt-induced pK_a shifts rule out a significant contribution of reverse protonation to the catalytic efficiency of the enzyme. At low salt concentration, the fraction of the catalytically active monoprotonated enzyme in the reverse protonated form (i.e., proton on the group with a pK_a of 3.7 and dissociated from the group with a pK_a of 7.2) is very small (≈ 0.03%). At higher salt concentrations, where the two pK_as become closer, the fraction of the monoprotonated enzyme in the reverse protonated form increases over 300-fold. However, there is no increase in the intrinsic reactivity, (k_cat/K_m)^lim, of the monoprotonated species. For other enzymes which may show such salt-induced pK_a shifts, this provides a convenient test for the role of reverse protonation.     

Leaf production and shoot dynamics of Thalassia testudinum by a direct census method

The annual leaf growth and shoot dynamics of Thalassia testudinum were examined in a meadow located near Havana City, Cuba, using direct censuses between January 1995 and January 1996. The net rate of shoot population change, specific shoot recruitment and mortality rates were calculated as the difference between the densities of shoots (tagged or untagged) in consecutive sampling events. The leaf biomass, the daily production, the turnover rate and the rate of leaf biomass loss were also estimated. The estimated mean dry leaf biomass (124.9 ± 9.5 g m⁻²), daily dry leaf production (3.3 ± 0.2 g m⁻² day⁻¹) and turnover rate (2.7 ± 0.1% day⁻¹) were comparable to values previously reported for this species in Cuba and elsewhere. The production of leaves and shoots were higher in spring, declined towards mid summer, and showed the minimum values in January. Shoot recruitment prevailed over shoot mortality from January to March and from July to August, whereas most of the annual shoot mortality occurred between May and July and between August and October. The meadow examined was in close demographic balance along the study period. The results demonstrate that direct census provides reliable estimates of rapid shoot dynamics in T. testudinum.     

Influence of level of zilpaterol chlorhydrate supplementation on growth performance and carcass characteristics of feedlot lambs

Forty-eight Pelibuey × Katahdin (38.8 ± 0.67 kg) crossbred male lambs were used in a 32-day feeding trial (four pens per treatment in a randomized complete block design), to evaluate the influence of zilpaterol (β₂-agonist) supplementation level on growth performance and carcass characteristics. Lambs were fed a dry-rolled corn-based finishing diet (3.04 Mcal/kg of ME) supplemented with 0, 0.15, 0.20, or 0.25 mg/kg of live weight d⁻¹ zilpaterol (as zilpaterol chlorhydrate, Zilmax^®, Intervet México, México City). DM intake averaged 1.099 ± 0.042 kg/d and was not affected (P = 0.40) by treatments. Compared with control lambs, zilpaterol supplementation increased gain efficiency (15.8%, P < 0.03), apparent energy retention per unit DMI (10.9%, P = 0.03), and tended to increased daily gain (16%, P < 0.07) and total gain (17.7%, P < 0.08). Zilpaterol supplementation did not affect (P = 0.20) carcass weight, longissimus muscle area (LM), or fat thickness, but increased (2.3%, P = 0.04) carcass dressing percentage and reduced (36%, P < 0.01) kidney-pelvic fat. Increasing level of zilpaterol supplementation increased total weight gain (linear component, P < 0.05), gain:feed (linear component, P < 0.01), and dressing percentage (linear component, P < 0.02), and decreased (linear component, P < 0.01) kidney-pelvic fat. We conclude that zilpaterol supplementation enhances growth performance and dressing percentage in lambs in a manner comparable to that of cattle (greater muscle accretion, reduced body fat). Responses to zilpaterol was optimal when supplemented at 0.20 mg of zilpaterol/kg of live weight d⁻¹.     

The effect of nitrogen containing bisphosphonates,zoledronate and alendronate,on the production of pro-angiogenic factors by osteoblastic cells

Bisphosphonates (BPs) have been shown to influence angiogenesis. This may contribute to BP-associated side-effects such as osteonecrosis of the jaw (ONJ) or atypical femoral fractures (AFF). The effect of BPs on the production of angiogenic factors by osteoblasts is unclear. The aims were to investigate the effect of (1) alendronate on circulating angiogenic factors; vascular endothelial growth factor (VEGF) and angiopoietin-1 (ANG-1) in vivo and (2) zoledronate and alendronate on the production of VEGF and ANG-1 by osteoblasts in vitro. We studied 18 post-menopausal women with T score ⩽ −2 randomized to calcium/vitamin D only (control arm, n = 8) or calcium/vitamin D and alendronate 70 mg weekly (treatment arm, n = 10). Circulating concentrations of VEGF and ANG-1 were measured at baseline, 3, 6 and 12 months. Two human osteoblastic cell lines (MG-63 and HCC1) and a murine osteocytic cell line (MLO-Y4) were treated with zoledronate or alendronate at concentrations of 10⁻¹²–10⁻⁶ M. VEGF and ANG-1 were measured in the cell culture supernatant. We observed a trend towards a decline in VEGF and ANG-1 at 6 and 12 months following treatment with alendronate (p = 0.08). Production of VEGF and ANG-1 by the MG-63 and HCC1 cells decreased significantly by 34–39% (p < 0.01) following treatment with zoledronate (10⁻⁹–10⁻⁶ M). Treatment of the MG-63 cells with alendronate (10⁻⁷ and 10⁻⁶) led to a smaller decrease (25–28%) in VEGF (p < 0.05). Zoledronate (10⁻¹⁰–10⁻⁶ M) suppressed the production of ANG-1 by MG-63 cells with a decrease of 43–49% (p < 0.01). Co-treatment with calcitriol (10⁻⁸ M) partially reversed this zoledronate-induced inhibition. BPs suppress osteoblastic production of angiogenic factors. This may explain, in part, the pathogenesis of the BP-associated side-effects.     

Macrophyte decomposition in a surface-flow ammonia-dominated constructed wetland: Rates associated with environmental and biotic variables

Decomposition of senesced culm material of two bulrush species was studied in a surface-flow ammonia-dominated treatment wetland in southern California. Decomposition of the submerged culm material during summer months was relatively rapid (k = 0.037 day⁻¹), but slowed under extended submergence (up to 245 days) and during fall and spring sampling periods (k = 0.009–0.014 day⁻¹). Stepwise regression of seasonal data indicated that final water temperature and abundance of the culm-mining midge, Glyptotendipes, were significantly associated with culm decomposition. Glyptotendipes abundance, in turn, was correlated with water quality parameters such as conductivity and dissolved oxygen and ammonia concentrations. No differences were detected in decomposition rates between the bulrush species, Schoenoplectus californicus and Schoenoplectus acutus.     

Improved PCR performance and fidelity of double mutant Neq A523R/N540R DNA polymerase

We previously reported that Neq A523R DNA polymerase is more efficient in PCR than wild-type Neq DNA polymerase, and amplifies products more rapidly. Neq A523R DNA polymerase also amplifies templates more rapidly than Pfu DNA polymerase, but has a lower fidelity than Pfu DNA polymerase. To improve product yield and the fidelity of amplification simultaneously, we constructed and characterized the double mutant Neq A523R/N540R. The yield of PCR products was greater for Neq A523R/N540R DNA polymerase than wild-type and other mutant DNA polymerases, and the Neq double mutant catalyzed amplification of a 12-kb PCR product from a lambda template with an extension time of 3 min. The PCR error rate of Neq A523R/N540R DNA polymerase (6.3 × 10⁻⁵) was roughly similar to that of Pfu DNA polymerase (4.8 × 10⁻⁵), but much lower than those of wild-type Neq DNA polymerase (57.2 × 10⁻⁵), Neq A523R DNA polymerase (13.1 × 10⁻⁵), and Neq N540R DNA polymerase (37.7 × 10⁻⁵). These results indicated that A523R and N540R mutations of Neq DNA polymerase had synergistic effects on its fidelity.     

First evidence of cell deformation occurrence during a Dinophysis bloom along the shores of the Gulf of Tunis (SW Mediterranean Sea)

Never before observed or cited in Dinophysis studies, deformations in Dinophysis acuminata and Dinophysis sacculus are reported throughout their cellular division phases (cytokinesis, and sulcal list regeneration) in 5 in situ cell cycle studies in the Punic harbors of Carthage (northern Tunisia). Two types of deformation were observed: invaginations in the ventral and dorsal margin and protuberances at the base of the left sulcal list. No virus or bacteria were detected with Syber green stain. In situ division rates (μ) varied among seasons and stations for the same species. D. acuminata exhibited moderate (0.22 day⁻¹) to high (0.68 day⁻¹) μ rates which were however very low (0.02–0.17 day⁻¹) for D. sacculus in autumn and moderate (0.21–0.35 day⁻¹) in late spring. In 2009 the seasonal distribution of Dinophysis indicates maximum Dinophysis cf. ovum abundance in March and a high number of D. acuminata in early June, while in 2010 maximum abundance of the same species was found in mid-June.Molecular and genetic studies and staining with specific fluorescent strains should be addressed to hopefully explain these Dinophysis cell deformations during their in situ division.     

Effect of different levels of concentrate allowances on rumen fluid pH,nutrient digestion,nitrogen retention and growth performance of weaner lambs

This experiment was conducted for 90 d to assess the effect of feeding graded levels of concentrate allowance on rumen fermentation characteristics, performance and nutrient utilisation of weaner lambs on restricted or high concentrate allowance using 60 weaner lambs of initial average live weight of 13.90 kg BW in a randomized design. The experimental treatments were 15 or 25 g kg⁻¹ BW or ad libitum concentrate allowance. Roughage source which contained Khejri (Prosopis cineratia) and Siris (Albizia lebback) leaves in 50:50 ratio was offered ad libitum to all the animals. Lambs supplemented with 15 g or ad libitum concentrate had similar dry matter intake (4.2 kg/100 kg BW) but significantly (p < 0.01) lower than 25 g concentrate supplemented group (4.9 kg/100 kg BW). Organic matter and CP intakes increased with increasing concentrate supplementation. Apparent digestibilities of dry matter, organic matter, CP, NDF, ADF and cellulose were significantly (p < 0.01) higher in ad libitum concentrate supplemented than 15 and 25 g concentrate supplemented lambs. Daily ME intake was significantly (p < 0.05) higher in 25 g and ad libitum concentrate supplemented lambs while ME intake kg⁻¹ gain was lower in ad libitum concentrate supplemented lambs (57 MJ kg⁻¹ gain) than those supplemented with 15 or 25 g concentrate (91 MJ kg⁻¹ gain). Generally, average daily gain increased with increasing levels of concentrate supplementation. Ad libitum concentrate supplemented lambs had significantly (p < 0.01) higher daily gains (151 g) than 15 and 25 g concentrate supplemented lambs (77 and 98 g, respectively). Feed efficiency was similar for 15 and 25 g concentrate supplemented lambs but significantly (p < 0.01) lower than the ad libitum concentrate supplemented lambs. All animals were in positive N-balance and the N-balance increased with increasing concentrate supplementation. Mean rumen fluid pH was significantly (6.6, p < 0.01) lower in ad libitum concentrate supplemented lambs compared to 15 or 25 g concentrate fed lambs (6.9). Rumen NH₃-N and total-N-concentrations peaked at 3 h post-feeding. Optimum rumen fluid pH, better nutrient digestibilities, higher N-retention improved growth by 49% of ad libitum concentrate fed lambs.