An improved method for a rapid determination of phytase activity in animal feed

The current direct colorimetric assay for phytase activity in feeds has interference from high P background and other factors. Our objective was to develop a rapid and reliable spin column method to accurately determine phytase activity in feed ingredients or complete diets. After the feed sample was extracted by stirring in 0.2 M citrate buffer, pH 5.5, for 30 min at room temperature, the oily layer of the supernatant fraction was removed by passing through an acrodisc syringe filter (0.45-microm HT Tuffryn membrane, Gelman Laboratory, Ann Arbor, MI). The filtrate was then loaded onto a spin column (MW cutoff 30,000, Millipore, Bedford, MA) to remove free phosphate before the phytase activity assay. Compared with the direct assay, this new procedure improved both accuracy and reproducibility. When diets contained phytase at 0 to 1,500 U/kg (as fed), the CV for multiple assays of the same samples (n = 6) by the new method ranged from 1 to 6% compared with 28 to 39% by the direct method. A linear relationship was found between the added phytase activity in practical diets and the analyzed activity by the new method (r2 = 0.99; P < 0.01). In conclusion, the spin column method is an improved assay for phytase activity in animal feed, and may be used for quality control of phytase supplementation.     

Identification of isolate‐specific resistance QTLs to phytophthora root rot using an intraspecific recombinant inbred line population of pepper (Capsicum annuum)

Quantitative trait loci (QTL) for resistance to phytophthora root rot caused by Phytophthora capsici were investigated using two Korean P. capsici isolates and 126 F₈ recombinant inbred lines derived from a cross of Capsicum annuum line YCM334 (resistant parent) and local cv. Tean (susceptible parent). The experimental design was a split plot with two replications. Highly significant effects of pathogen isolate, plant genotype, and genotype × isolate were detected. QTL mapping was performed using a genetic linkage map covering 1486·6 cM of the pepper genome, and consisted of 249 markers including 136 AFLPs (Amplified Fragment Length Polymorphisms), 112 SSRs (Simple Sequence Repeats) and one CAPS (Cleaved Amplified Polymorphic Sequence). Fifteen QTLs were detected on chromosomes 5 (P5), 10 (P10), 11 (P11), Pb and Pc using two data processing methods: percentage of wilted plants (PWP) and relative area under the disease progress curves (RAUDPC). The phenotypic variation explained by each QTL (R²) ranged from 6·0% to 48·2%. Seven QTLs were common to resistance for the two isolates on chromosome 5 (P5); six were isolate‐specific for isolate 09‐051 on chromosomes 10 (P10) and Pc, and two for isolate 07‐127 on chromosomes 11 (P11) and Pb. The QTLs in common with the major effect on the resistance for two isolates explained 20·0–48·2% of phenotypic variation. The isolate‐specific QTLs explained 6·0–17·4% of phenotypic variation. The result confirms a gene‐for‐gene relationship between C. annuum and P. capsici for root rot resistance.     

Plant invasion and management in turf‐dominated waste landfills in South Korea

A floristic survey was conducted in turf‐dominated fields on some waste landfills in South Korea, to quantify the degree of plant invasion in those degraded lands. The dominant species, except turf (Zoysia japonica), were Conyza canadensis, Setaria viridis, Digitaria sanguinalis and Aster tripolium. Mean species number declined linearly with distance from the edge of the turf stands. Biomass of each species ranged between 0.1 g and 122.6 g, decreasing linearly with distance from the edge of the stand and declining exponentially with turf coverage. Mean soil moisture storage and organic matter content were 12.59 per cent and 5.81 per cent, respectively. Sum of relative cover, frequency and biomass of all species with environmental variables (distance from the stand edge, turf cover, soil moisture storage, soil organic matter content and the ratio of species number of exotic plants to that of native plants) were ordinated by DCA (detrended correspondence analysis). The first axis of the DCA was positively correlated with distance from the stand edge and turf cover and the second axis of the DCA was negatively correlated with soil organic matter content. Highly and lowly invasive species were indicated from DCA results. From analysis of dispersal type, barochore and anemochore were the most widespread of other dispersal types but it was suggested that anthropochore played an important role in plant invasion in these degraded wastelands. Establishment of an edge transition zone, enhancement of turf cover and reduction in traffic is recommended as management options for turf fields. Copyright © 2002 John Wiley & Sons, Ltd.     

Development of a polymerase chain reaction (PCR) procedure for the detection of baculovirus associated with white spot syndrome (WSBV) in penaeid shrimp

The causative viral agent was purified from diseased shrimp Penaeus japonicus with white spot syndrome (WSBV). Several hundred clones were obtained from libraries of the purified viral genomic DNA. According to the results of nucleotide sequence analysis, none of the WSBV clones showed considerable sequence homology with those of other known viruses, indicating that WSBV is a new virus causing a serious disease in shrimp. Based on the sequence data of WSBV genomic DNA, a pair of polymerase chain reaction (PCR) primers was designed. After 30 cycles of PCR amplification of viral genomic DNA extracted from WSBV, a single product of the expected size was detected. Southern blot hybridization confirmed that the amplified product was specific to the DNA of WSBV. The PCR system was able to detect 1 pg of WSBV DNA after 30 cycles, and efficiently amplify the target region of WSBV gene in the total nucleic acids extracted either from the diseased shrimp or hatchery shrimp with no signs of viral infection.     

Successful treatment of gastrocnemius muscle rupture in a long-tailed goral rescued from a live trap: a case report

We describe a case of gastrocnemius muscle rupture in a goral that got caught in a live trap set up to capture wild boars. We established a diagnostic method based on clinical signs and imaging examination findings, including magnetic resonance imaging for gastrocnemius muscle rupture with symptoms such as hindlimb lameness and plantigrade posture of unknown cause in wild animals. Although this condition is rare, this report will help veterinarians diagnose several diseases of the musculoskeletal and nervous systems in wild animals. This is the first report of successful diagnosis and treatment of gastrocnemius muscle rupture in gorals in Korea.     

群体饲喂条件下猪个体采食量的评估模型

在大多数生长试验中,试验动物往往成群地饲养在同一圈中。在试验过程中,有时有个别动物在生长速度上会表现出与其同伴较大的差异,或者死亡。出现此种情况时,如果不能为统计日平均采食量(average daily feed intake,ADFI)和饲料效率而对试验动物的圈采食量(penfeedintake,PFI)进行重新评估,那么试验数据集将丢弃该圈的观察值。因此,我们开发了一个模型,用来评估群体饲喂条件下猪的个体采食量(Individual feed intake,IFI),并通过群体饲喂模拟试验对该评估模型进行验证。在推荐模型中,将每个受影响圈的采食量(FeedIntake,FI)分为用于维持生存需要的采食量(FIm)和用于生长需要的采食量(FIg)两个部分。首先,试验期间个体猪的FIm按NRC(1998)所列出的维持生存所需代谢能的评估方法进行测算,然后将同圈内所有猪的FIm相加,即得到圈FIm。圈采食量(PFI)与圈FIm之差就是该圈动物用于生长所需的那部分采食量。接下来,把该差额部分的采食量平均分给圈内动物每单位增重上,即为FIg。最后,猪圈中被淘汰猪的IFI估测值即为所淘汰猪的FIm与FIg之和,原来的圈采食量(PFI)减去该FI估测值后即得剩余猪的新PFI。个体采食量评估的准确性取决于维持生存代谢能预测方程和饲料原料能量分析值的准确性。在模拟试验中,我们将推荐模型的精确度与2个其它模型的精确度作了比较。模拟试验1显示,在所有测试阶段中推荐模型的精确度至少高于其它2个模型中的任一种方法(P<0.001);模拟试验2也证明,推荐模型的精确度高于其他2种方法。由于IFI的人工计算相对麻烦,为此我们开发了饲料采食量校正电子数据表(feed intake correction spreadsheet,FICS),即一个利用宏指令可对FI进行校正的Excel电子数据表。FISC包括了推荐模型中的所有测算程序,用户可免费从指定的网页上下载Excel文件和使用说明书。     

Implication of Echinochrome A in the Plasticity and Damage of Intestinal Epithelium

The diverse therapeutic feasibility of the sea urchin-derived naphthoquinone pigment, Echinochrome A (Ech A), has been studied. Simple and noninvasive administration routes should be explored, to obtain the feasibility. Although the therapeutic potential has been proven through several preclinical studies, the biosafety of orally administered Ech A and its direct influence on intestinal cells have not been evaluated. To estimate the bioavailability of Ech A as an oral administration drug, small intestinal and colonic epithelial organoids were developed from mice and humans. The morphology and cellular composition of intestinal organoids were evaluated after Ech A treatment. Ech A treatment significantly increased the expression of LGR5 (~2.38-fold change, p = 0.009) and MUC2 (~1.85-fold change, p = 0.08). Notably, in the presence of oxidative stress, Ech A attenuated oxidative stress up to 1.8-fold (p = 0.04), with a restored gene expression of LGR5 (~4.11-fold change, p = 0.0004), as well as an increased expression of Ly6a (~3.51-fold change, p = 0.005) and CLU (~2.5-fold change, p = 0.01), markers of revival stem cells. In conclusion, Ech A is harmless to intestinal tissues; rather, it promotes the maintenance and regeneration of the intestinal epithelium, suggesting possible beneficial effects on the intestine when used as an oral medication.     

Serum concentrations of leptin and adiponectin in dogs with chronic kidney disease

BackgroundAn imbalance in adipokines is associated with the progression of chronic kidney disease (CKD) in humans. However, alterations in adipokines in dogs with CKD remain unclear.ObjectivesTo examine whether adipokine concentrations in serum differ between healthy dogs and dogs with CKD and to determine the correlation between serum adipokine concentrations and CKD severity in dogs.AnimalsTwenty dogs with CKD and 10 healthy dogs.MethodsIn this cross‐sectional study, serum concentrations of leptin, adiponectin, interleukin (IL)‐6, IL‐10, IL‐18, and tumor necrosis factor (TNF)‐α were measured in healthy dogs and dogs with CKD, which were classified according to the International Renal Interest Society guidelines.ResultsSerum leptin concentrations were positively correlated with systolic arterial blood pressure (r = .41), creatinine concentrations (r = .39), and symmetric dimethylarginine concentrations (r = .73). Serum adiponectin concentrations (median [range]) in CKD dogs with borderline or non‐proteinuric (20.25 [14.9‐45.8] ng/mL) were significantly higher than those in proteinuric CKD dogs (13.95 [6.4‐22.1] ng/mL; P = .01). Serum IL‐6 (median [range]; 43.27 [24.30‐537.30] vs 25.63 [6.83‐61.03] pg/mL; P = .02), IL‐18 (median [range]; 25.98 [11.52‐280.55] vs 10.77 [3.53‐38.45] pg/mL; P = .01), and TNF‐α (median [range]) concentrations (11.44 [8.54‐38.45] vs 6.105 [3.97‐30.68] pg/mL; P = .02) were significantly different between proteinuric and borderline or non‐proteinuric CKD dogs.Conclusions and Clinical Importanceleptin and adiponectin concentrations in serum might be associated with severity of CKD and proteinuria in dogs with CKD, respectively.