Evaluation of ingredient combinations from differing origins (fishmeal,terrestrial animal and plants) and two different formulated nutrient targets on rainbow trout growth and production efficiency

A 12‐week trial was performed with rainbow trout (Oncorhynchus mykiss) utilizing combinations of ingredients at two nutrient targets. Ingredient combinations were fishmeal‐based diet (FMD), animal product‐based diet (APD), plant product‐based diet (PPD), novel plant‐based diet (NPD) and potential future plant‐based diet (FPD). Two nutrient concentrations were targeted: 1) to meet published amino acid targets for rainbow trout utilizing approximately 450 g/kg crude protein (400–420 g/kg digestible protein) and 2) to meet the amino acid targets based on ideal amino acid balance of trout muscle for Lys, Met and Thr utilizing approximately 400 g/kg crude protein (370–380 g/kg digestible protein). Interactive effects between ingredient combination and nutrient concentration occurred across all response variables. When diets were formulated to Target 1, fish consuming FMD and APD displayed better weight gain and feed conversion ratio (FCR) than plant‐based diet, while graded effects were found within the plant‐based treatments. When differing ingredient combinations were utilized and formulated to Target 2, fish grew equally well except for the NPD treatment which supported lower growth and higher FCR. Based on the data from the current experiment, one can completely remove fishmeal from trout feeds and reduce protein levels when dietary digestible amino acids are balanced.     

Calibration and validation of optical chlorophyll‐measuring devices for use in predicting crude protein concentration in tropical grass herbage

The objective of this experiment was to evaluate the Fieldscout CM 1000 NDVI and Yara N‐Tester as easy‐to‐use and cost‐effective tools for predicting foliar chlorophylls (a, b and total) and crude protein (CP) concentrations in herbage from three tropical grass species. Optical chlorophyll measurements were taken at three stages (4, 8 and 12 weeks) of regrowth maturity in Guinea grass (Panicum maximum) and Mulato II (Brachiaria hybrid) and at 6 and 12 weeks maturity in Paspalum spp (Paspalum atratum). Grass samples were harvested subsequent to optical measurements for laboratory analysis to determine CP and solvent‐extractable chlorophylls (a, b and total) concentrations. Optical chlorophyll measurements and CP concentrations were highly correlated (Yara N‐Tester: r² = 0·77–0·89; Fieldscout CM 1000 NDVI: r² = 0·52–0·84). Crude protein prediction models from the Yara N‐Tester and Fieldscout CM 1000 NDVI accounted for 70–89% and 44–73% CP variability, respectively, in Mulato II and Guinea grass. The Yara N‐tester produced more accurate and reliable CP estimates based on very high concordance correlation coefficient [CCC (0·73–0·91)] and low rMSPE, mean and regression bias. It is concluded that the Yara N‐Tester produces more accurate and reliable CP estimates of tropical pastures.     

Using near‐infrared reflectance spectroscopy to predict the digestible protein and digestible energy values of diets when fed to barramundi,Lates calcarifer

This study examined the potential of using near‐infrared spectroscopy (NIRS) to predict nutrient digestibility parameters (digestible protein and digestible energy) of compound diets when fed to barramundi. A series of 60 diets were assessed for their protein and energy digestibilities in a series of five experiments over a 5‐year period from 2009 to 2014. Considerable variance was observed in the digestibility parameters of diets across the experiments, providing a suitable range in diet digestible protein and digestible energy values from which to develop a NIRS calibration. Samples of the same diets were also scanned using a diode array near‐infrared spectrophotometer (DA‐NIRS). The spectra were obtained by the DA‐NIRS and were chemometrically calibrated against the digestible value data using multivariate analysis software. The results in terms of standard error of cross‐validation (SECV), residual prediction deviation (RPD) and correlation coefficient (R²) show good relationships (R² > 0.8) between the predicted and observed parameters for both the digestible protein and digestible energy parameters assessed. This study therefore demonstrates that it is possible to use NIRS technology to provide rapid estimates of the digestible protein and digestible energy values of compound diets for barramundi in near real time.     

草鱼鳃组织蛋白双向电泳技术的建立及优化

为建立并优化草鱼鳃蛋白质组双向电泳的技术体系,实验将草鱼鳃经裂解处理后,通过固相p H梯度胶条进行一向等电聚焦和SDS聚丙烯酰胺凝胶垂直电泳进行蛋白分离,对不同的裂解液配方、蛋白纯化方法、IPG胶条的分离范围、上样量和染色方法等进行了探索和优化,并应用lmage Master 2D Platinum 7.0软件对电泳图谱进行了分析。结果表明:采用裂解液中添加DTT来取代Tris和TBP、选择分离范围为pH4-7的IPG胶条和150μg的主动水化上样,电泳结束后对凝胶进行硝酸银染色,获得了覆盖范围广、低背景、高分辨率、适合差异蛋白分析的双向电泳参考图谱。     

Responses of glutamine synthetase activity and gene expression to nitrogen levels in winter wheat cultivars with different grain protein content

Glutamine synthetase (GS) plays a central role in plant nitrogen (N) metabolism, which improves crops grain protein content. A pot experiment in field condition was carried out to evaluate GS expression and activity, and grain protein content in high (Wanmai16) and low grain protein (Loumai24) wheat cultivars under two N levels (0.05 and 0.15 g N kg⁻¹ soil). High nitrogen (HN) resulted in significant increases in GS1 and GS2 expression at 10 days after anthesis (DAA), and higher GS activity during the entire grain filling stage. HN also significantly increased yield, grain protein content and protein fraction (except for glutenin of Luomai24) in two wheat cultivars, which indicated that it increased grain yield and protein content by improving nitrogen metabolism. Wanmai16 showed higher grain protein content, gliadin and glutenin content, and had higher expression level of GS2 both in flag leaves and grains at early grain filling stage. However, Luomai24 had greater yield and higher expression level of GS1. The difference expression of GS2 and GS1 genes indicates they had various contributions to the accumulation of protein and starch in wheat grains, respectively. The results suggest that GS2 would be serving as a potential breeding target for improving wheat quality.     

Evaluation of dietary inclusion of housefly maggot (Musca domestica) meal on growth,fillet composition and physiological responses for barramundi,Lates calcarifer

The study was to evaluate the effects of dietary fish meal (FM) partially replaced by housefly maggot meal (HMM) on growth, fillet composition and physiological responses of juvenile barramundi, Lates calcarifera. HMM at 100, 150, 200 and 300 g kg^?1 was supplemented in the basal diet to replace dietary FM protein. Basal diet without HMM supplementation was used as control. Total of five experimental diets were fed to triplicate groups of juvenile barramundi (initial weight: 9.66 ± 0.22 g) in a flow‐through rearing system for 8 weeks. Fish fed all experimental diets showed no effects (P > 0.05) on weight gain and whole body protein, lipid and moisture content. Fish fed control diet and 100 g kg^?1 HMM diet had the highest (P < 0.05) hepatic superoxide dismutase (SOD) activity, followed by 150 g kg^?1 HMM group, the lowest in 200 and 200 g kg^?1 HMM groups. Hepatic thiobarbituric acid reactive substance (TBARS) value was the highest in fish fed 150–300 g kg^?1 HMM diets, followed by 100 g kg^?1 HMM group and the lowest in fish fed the control diet. Fish fed the 300 g kg^?1 HMM diet had lower plasma lysozyme activity than fish fed other diets. The results indicated that up to 300 g kg^?1 HMM can be used to substitute dietary FM protein without negative effect on growth. Although physiological responses were also considered, up to 100 g kg^?1 HMM in barramundi diet was recommended.     

Seasonal variations of immune parameters in the coelomic fluid of sea cucumber Apostichopus japonicus cultured in pond

In order to understand the effects of seasonal change on the immunity of sea cucumber Apostichopus japonicus cultured in pond, A. japonicus with body weight of 12.2 ± 4.5 g (sample A) and 32.6 ± 7.1 g (sample B), respectively, were collected monthly and randomly from a typical pond during a year cycle and employed for the evaluation of immunocompetence. Simultaneously, the environmental factors in the pond including water temperature, pH, salinity and dissolved oxygen (DO) were measured using a handheld multiparameter meter. The activities of acid phosphatase (ACP), alkaline phosphatase (ALP), lysozyme (LYZ), phenoloxidase (PO), superoxide dismutase (SOD), catalase (CAT) and myeloperoxidase (MPO) in the coelomic fluid were selected for the evaluation of A. japonicus immunocompetence and determined using biochemical methods. The results showed that in both samples, the activities of all determined enzymes had small values in winter and early spring, and LYZ, CAT and MPO activities also presented small values in summer, suggesting that pond‐cultured A. japonicus underwent immunosuppression twice during a year cycle, and the immunosuppression occurred in winter and early spring was more severe than that occurred in summer. In addition, most of the determined enzymes showed different variations between the two samples, and had significantly negative correlation with protein concentration, which was significantly and negatively correlated with water temperature, revealing that body weight and water temperature might have crucial effects on the immunity of A. japonicus cultured in pond.     

Effects of dietary protein and lipid levels with different protein‐to‐energy ratios on growth performance,feed utilization and body composition of juvenile red‐spotted grouper,Epinephelus akaara

An 8‐week feeding trial was conducted to assess dietary protein and lipid levels on growth performance, feed utilization and body composition of juvenile red‐spotted grouper (7.85 ± 0.03 g fish^?1). Nine semi‐purified diets were formulated containing varying protein levels (440–520 g kg^?1, dry matter) and lipid levels (60–120 g kg^?1, dry matter). The weight gain of juvenile Epinephelus akaara was affected by dietary protein (p = .005) and its interaction with dietary lipid (p = .020). Viscerosomatic index, intraperitoneal fat ratio and whole‐body lipid level increased with increasing dietary lipid level (p < .001). Nitrogen retention was not affected by dietary protein and lipid, while lipid retention decreased with increasing dietary lipid level (p < .001). The plasma blood urea nitrogen increased with increasing dietary protein level (p = .003). This study showed that diet with 520 g kg^?1 protein and 60 g kg^?1 lipid with 30.58 mg kJ^?1 P:E provided a maximal growth for this species. Moreover, an increase in dietary lipid levels (from 60 to 90 g kg^?1) could reduce the protein requirement (from 520 to 480 g kg^?1) without affecting the growth performance, while higher fat deposition was observed in fish fed high‐lipid diets.     

一个水稻小热休克蛋白的异源表达及寡聚特性分析

【目的】在前期研究中,本实验室已克隆了一个水稻小热休克蛋白基因(OsSHSP17.6),并发现该基因的表达明显受到热激和病毒侵染调控,表明该蛋白可能在逆境胁迫过程中起重要作用。本研究的目的在于进一步明确OsSHSP17.6的特性。【方法】在本研究中,进一步将该基因亚克隆至原核表达载体p ET-32a并导入大肠杆菌E.coli BL21(DE3)pLysS诱导表达,通过亲和层析的方法纯化了该重组蛋白,进一步用于非变性聚丙烯酰胺凝胶电泳和Western blotting分析。【结果】异源表达的重组OsSHSP17.6能减轻IPTG对宿主菌的毒害。非变性聚丙烯酰胺凝胶电泳和Western blotting分析显示纯化的重组OsSHSP17.6蛋白在体外能形成同源二聚体和寡聚体。【结论】这些结果支持OsSHSP17.6是一个有功能的分子伴侣蛋白并表明该蛋白可能通过形成同源寡聚体的方式参与逆境胁迫反应,这为进一步明确OsSHSP17.6的功能机制奠定基础。     

Outbreak of anaplasmosis associated with novel genetic variants of Anaplasma marginale in a dairy cattle

During early lactation, dairy cows may present a transient immunosuppressive state and develop anaplasmosis caused by Anaplasma marginale. In this study, clinical anaplasmosis in dairy cattle in the Thrace region of Turkey was investigated with respect to within-herd prevalence, vertical transmission, and genetic diversity. In March and September 2015, thirty lactating cows showed primary clinical signs of anaplasmosis, including fever, anaemia, decreased milk yield, anorexia, and laboured breathing. Symptoms disappeared in most cows after administration of long-acting oxytetracycline, but nine of them (30%) died. Following diagnosis based on clinical signs, microscopy and molecular findings, blood samples were collected from apparently healthy lactating cows (n = 184), pregnant heifers (n = 39) and newborn calves (n = 24). DNA was extracted from each sample and analyzed for the presence of major surface proteins (MSPs) of A. marginale, followed by sequencing to assess diversity of isolates. Microscopic examination of erythrocytes revealed A. marginale inclusion bodies in symptomatic cows. Examination of thin blood smears showed 3.8% of the lactating, clinically asymptomatic, cows to be infected with A. marginale, while nPCR detected 31.0% positive. A. marginale infection was not detected in pregnant heifers by either method. Congenital infection was found in one calf by nPCR. This is the first report of transplacental transmission of A. marginale in Turkey. The MSP4 sequence analyses showed high genetic diversity among the isolates, presenting 97.6-99.6% homology at the amino acid level. The sequences of MSP1a amplicons revealed genetic diversity providing three new tandem repeats.