Effects of Confluent, Roscovitine Treatment and Serum Starvation on the Cell-cycle Synchronization of Bovine Foetal Fibroblasts

The present study was designed to examine the effects of cell-cycle synchronization protocols, such as confluent, roscovitine treatment and serum starvation, in bovine foetal fibroblasts on synchronization accuracy at G0/G1, viability, apoptosis, necrosis and ploidy for use as a nuclei donor. The cells in 5-10 passages were randomly allocated into three treated groups. Cells were cultured either in Dulbecco's modified Eagle's medium (DMEM) + 10% foetal bovine serum (FBS) until 90% confluent (group 1, confluent), in DMEM + 10% FBS + 30 microM roscovitine for 12 h (group 2, roscovitine), or in DMEM + 0.5% FBS for 5 days (group 3, serum starvation). Most of the cells (>80%) in all groups were arrested at the G0/G1 stage. Although the rates did not differ, cells in group 1 showed an increased cell population arrested at the G0/G1 phase. Significantly (p < 0.05) higher rates of apoptosis occurred in group 3 than in group 1 and 2 (10% vs 6% and 6%, respectively). No differences in chromosomal abnormality were observed among groups. However, by increasing the number of cell culture passages up to 15, significantly (p < 0.05) higher chromosomal abnormality was observed than in 5 and 10 passages (39% vs 28% and 23%, respectively) in group 1. The results clearly indicated that bovine foetal fibroblasts could be effectively synchronized at G0/G1 stages by all the three different treatments, confluent, roscovitine and serum starvation. However, cells in confluent showed reduced apoptosis and necrosis when they underwent 5-10 passages, exhibiting increased percentage of cells with stable chromosome diversity. Hence, cells in confluent merit further studies before they could be used as nuclear donors.     

APEX model simulation of runoff and sediment losses for grazed pasture watersheds with agroforestry buffers

Buffers have been found to reduce non-point source pollution (NPSP) from watersheds. Hydrologic simulation models assist in predicting the effects of buffers on runoff and sediment losses from small watersheds. The objective of this study was to calibrate, validate and simulate runoff and sediment losses and compare buffer effects on NPSP losses relative to control watersheds (no buffer) for seven years. The experimental design consists of four watersheds under pasture management which were monitored from 2002 through 2008; two with agroforestry buffers (AgB 100 and AgB 300) and two control watersheds (CW 400 and CW 600). Pasture areas included red clover (Trifolium pretense L.) and lespedeza (Kummerowia stipulacea Maxim.) planted into fescue (Festuca arundinacea Schreb.) while the agroforestry buffer area included Eastern cottonwood trees (Populus deltoids Bortr. ex Marsh.) planted into fescue. The APEX model was calibrated from 2002 to 2005 and was validated from 2006 to 2008. The r ² and NSE values for the calibration and validation periods of the runoff varied from 0.52 to 0.78 and 0.50 to 0.74, respectively. The model did not predict sediment loss very well probably due to insufficient number of measured events and low measured sediment loss. The measured runoff was 57% higher for CW watersheds compared to AgB watersheds. The measured sediment loss was 95% higher for CW watersheds compared to AgB watersheds. After calibrating and validating the model, it was run for long-term scenario analyses for 10 years from 1999 to 2008. Simulated buffer width had a significant influence on runoff. Simulated runoff decreased by 24% when the buffer width was doubled compared to losses associated with the measured buffer width. Simulated runoff from the CW watersheds was 11% higher with double stocking density (relative to measured density) compared to AgB watersheds with double stocking density. With half stocking density (relative to measured density), the AgB watershed had 18% lower runoff compared to CW. Results from this study imply that establishment of agroforestry buffers on grazed pasture watersheds reduce runoff and sediment losses compared to control watersheds without buffers.     

Molecular diversity of barley yellow dwarf virus-PAV from China and the Czech Republic

Wheat yellow dwarf disease (BYD), caused by different species of barley/cereal yellow dwarf viruses (B/CYDVs), is one of the most serious cereal diseases in China and the Czech Republic. Because genetic diversity of the virus directly influences disease epidemiology, the molecular diversity and population structure of 24 Chinese isolates and 16 the Czech Republic isolates of BYDV-PAV from different regions in two countries were analyzed by sequencing their coat protein (CP) and readthrough protein (RTP) domain (RTD) genes and comparing the sequences with six CP and 16 RTP sequences of BYDV-PAV isolates from the NCBI database based on nucleotide identity position, phylogenetic analysis and nucleotide diversity. Nucleotide identities between the Chinese and the Czech Republic isolates for the CP were 76.6–99.4%, 73.9–89.1% for RTD (ORF5), respectively. The Chinese and the other country isolates showed 74.7–99.2% nucleotide identity for RTP (ORF3+ORF5). Phylogenetic analysis of CP sequences showed that 20 Chinese isolates clustered in the same clade, but the other four Chinese isolates clustered in another clade with the isolates from the Czech Republic and other counties. The population of BYDV-PAV in China had greater nucleotide variability and was more divergent than that in the Czech Republic. Geographical and ecological factors but not hosts might contribute to the population differences in the two countries.     

Thermochemistry and aqueous solubilities of hydrotalcite-like solids

Hydrotalcites are used in technology as catalysts and anion exchangers and are important sinks for environmental contaminants. Their compositional variability makes it important, but difficult, to estimate their aqueous solubility. We report calorimetric measurements of the heats of formation of cobalt-aluminum hydrotalcite phases. The heat and free energy of formation from the elements are equal to those of mechanical mixtures of binary compounds, namely hydroxides and carbonates. The interlayer anion is much more important than the cation in determining the solubility of the hydrotalcite phase and its ability to contain or release heavy metals to the environment. Because hydrotalcites do not have an unreactive polymer as a structural core, their aqueous stability will change dramatically with composition, particularly with anion content. This simple mechanical mixture model allows prediction of aqueous solubilities and trace metal retention in a variety of geochemical settings.     

Effect of substituting live feed with formulated feed on the reproductive performance and fry survival of Siamese fighting fish, Betta splendens (Regan, 1910)

This study aimed to elucidate the effect of partial or complete replacement of live feed (LF) (Tubifex) with formulated feed (FF) on the reproductive performance of Betta splendens. Three hundred B. splendens fry (average weight 0.19 ± 0.01 g) were equally distributed into five different groups, each with three replicates. They were fed for 105 days with following different diets: control (C)—100% LF; T1—75% LF, 25% FF; T2—50% LF, 50% FF; T3—25% LF, 75% FF, and T4—100% FF. Results showed that the average number of hatched larvae (654 ± 101) and fry survival after 2 weeks of rearing (428 ± 70), after completion of three spawning, were recorded highest in the control group, which was, however, not significantly different from the T1, T2, and T3 groups. At the end of the feeding trial, the highest hatching percentage (90.3%) was registered in the T2 group, which was not significantly different from the control and T1 groups. The T2 group also recorded highest fry survival (65.54%) after completion of three spawning, which was not significantly (P < 0.05) different from the T1 and T3 groups. Control diet contained higher saturated fatty acid (63.23%) than formulated diet (29.80%). In the whole-body tissue, highest level of EPA (0.42%) and DHA (3.13%) were found in the T4 group followed by T3 group. The DHA/EPA ration was recorded highest in the T2 group (10.96%), which did not differ significantly from the T1 and T3 groups. Significant positive correlation was observed between saturated fatty acid levels in fish whole-body tissue and number of hatched larvae (Y = 30.81 × −825.3, R ² = 0.968) and fry survival after 2 weeks of rearing (Y = 21.38 × −580.9, R ² = 0.967). Considering all these factors, it can be concluded that the live feed Tubifex can be replaced up to 50% without any adverse effect on the reproductive performance and fry survival of B. splendens.     

Replacement of live feed by formulated feed: effect on the growth and spawning performance of Siamese fighting fish (Betta splendens,Regan, 1910)

Partial replacement of live feed (LF) with formulated feed (FF) was performed in Betta splendens to evaluate their growth, survival and reproductive performance. Three hundred B. splendens fry of uniform size (mean weight 0.19 g±0.01) were equally distributed in five treatment groups with each of three replicates in glass aquaria of 35 L capacity. Fish were offered diets of different ratio of LF and FF like T1 (control) – 100% LF; T2 – 75% LF, 25% FF; T3 – 50% LF, 50% FF; T4 – 25% LF, 75% FF and T5 – 100% FF. Highest (P<0.05) body weight gain (% BWG) (67.1±1.03) and specific growth rate (2.34±0.02) were observed in the T2 group. Best feed conversion ratio was also found in T2 (2.40±0.11), which was similar to T1 and T3. The highest survival rate (%) was recorded in T1, T2 and T3 groups (97±1.7), which was similar to T4 and the lowest in T5 (49±2.0). The highest gonadal weight (0.12 g±0.01) and gonadosomatic index (15.17±0.50) was observed in the T3 group during the first sampling after 63 days. No significant changes in spawning performance were observed in the second sampling after 21 days after first sampling. However, T4 group registered the highest fecundity and fertilized eggs at the end of 105 days of experiment. From the present study, it concludes that LF can be successfully replaced to the extent of 25% by FF without any detrimental effect on the growth, survival and spawning performance of B. splendens.     

用地形特征预测喜马拉雅某集水区径流

长期以来 ,科学家和工程师们都试图计算出不规则集水区的径流量。用于降雨—径流模型的最常用方法是通过线性理论来确定某盆地的单位瞬时水位曲线图 (ＩＵＨ)。最近 ,地形学技术已发展到可将径流与集水区地形特征联系起来。这些地形特征从集水区已有的拓扑关系图、空间图片等资     

Effect of dietary supplementation of periphyton on growth,immune response and metabolic enzyme activities in Penaeus monodon

A 60‐day indoor trial was conducted to study the effect of periphyton supplementation on metabolic and immune responses in tiger shrimp, Penaeus monodon. Periphyton developed over bamboo substrate in outdoor tanks (15 m²) was used as dietary supplement for P. monodon (2.02 ± 0.04 g) reared in 1000 L FRP tanks. Graded levels of periphyton were included in shrimp basal diets: 0% (P0), 3% (P3), 6% (P6), 9% (P9) and P0 diet with natural periphyton (NP) over bamboo substrate. At the end of the trial, P6 and NP showed significantly higher (P < 0.01) growth rate, 23.9% and 20%, respectively, compared with control, P0. Comparatively, lower level of metabolic enzymes, such as lactate dehydrogenase, malate dehydrogenase, aspartate aminotransferase and alanine aminotransferase, was recorded in treatments P3, P6 and NP compared with control, P0. The periphyton‐supplemented group, P3 had significantly higher (P < 0.05) superoxide dismutase (15.83 ± 0.96) and catalase activity (15.73 ± 0.69) compared to 6.88 ± 2.84 and 9.15 ± 0.67 unit mg^?1 protein min^?1_, respectively, in P0. Similarly, higher total haemocyte counts, 32.58 ± 1.30, 28.51 ± 3.12 and 27.26 ± 4.43 × 10⁶ cells mL^?1_, were recorded in P6, NP and P3, respectively, compared to P0, 23.57 ± 1.80 × 10⁶ cells mL^?1. After challenge with Vibrio harveyi, P3 recorded the highest relative percentage survival 67% followed by NP (58%) and P6 (42%) compared with control. However, treatment with highest periphyton inclusion (P9) did not differ significantly with P0 on growth and immunological parameters. This study indicates that periphyton supplementation at 3–6% level improves growth, immune response and metabolic activities in P. monodon.     

Metabolic and digestive enzyme activity of Pangasianodon hypophthalmus (Sauvage, 1878) fingerlings in response to alternate feeding of different protein levels in the diet

A feeding trial was conducted for 90 days to assess the metabolic and digestive enzyme activity in response to alternate or mixed feeding of different protein levels in the diet of Pangasianodon hypophthalmus fingerlings. One hundred and forty four fingerlings with an average weight of 5.0–5.5 g were distributed into four experimental groups each with triplicate. Four iso‐caloric diets with varying level of crude protein viz., 35%, 30%, 25% and 20% were made and designated as diet 35P, 30P, 25P and 20P respectively. Four treatments were maintained based on four different feeding schedules such as continuous feeding of diet 35P (35P/35P), alternate feeding of 1 day diet 35P next day diet 30P (35P/30P), alternate feeding of 1 day diet 35P next day diet 25P (35P/25P) and alternate feeding of 1 day diet 35P next day diet 20P (35P/20P). As a result, specific growth rate (SGR) and weight gain percentage were relatively higher in treatment (35P/25P) followed by (35P/35P) and (35P/30P). The metabolic enzymes such as lactate dehydrogenase (LDH), malate dehydrogenase (MDH), glucose‐6‐phosphate dehydrogenase (G6PDH), Glucose‐6‐phosphatase (G6Pase), alkaline phosphatase (ALP), acid phosphatase (ACP), alanine aminotransferase (ALT), aspartate aminotransferase (AST) were expressed better in (35P/35P) group followed by (35P/25P) and (35P/30P) groups. The digestive enzymes, protease and lipase activities were higher in (35P/35P) and (35P/25P) groups. The present study revealed that the better nutrient utilization and growth of 35P/25P group is because of their balanced digestive and metabolic enzyme activities through alternate level of higher and lower protein diet.