Differential levels of antioxidants in paraquat-resistant and -susceptible Erigeron canadensis biotypes in Korea

To characterize the biochemical differences in paraquat-resistant and -susceptible biotypes of Erigeron canadensis L. collected from Korea, we investigated the constitutive levels of various antioxidants such as antioxidant enzymes and low molecular weight antioxidants in leaves, as well as after paraquat treatment. The activities of superoxide dismutase, peroxidase, ascorbate peroxidase, and catalase were higher in the paraquat-resistant biotype than in the paraquat-susceptible biotype. Reduced ascorbic acid content was higher in the resistant biotype, but the content of reduced glutathione was higher in the susceptible biotype. These results indicate that one of the paraquat-resistant mechanisms in E. canadensis in the present study might be related to protecting the activities of antioxidant enzymes, such as superoxide dismutase, peroxidase ascorbate peroxidase, and catalase, as well as the contents of low molecular weight antioxidants such as ascorbate and glutathione.     

NO介导的Ca2+信号对渗透胁迫下紫花苜蓿幼苗光合特征及抗性的影响

以紫花苜蓿幼苗为材料,用聚乙二醇(PEG-6000)作为渗透介质人工模拟干旱条件,外源喷施NO供体硝普钠(SNP)、钙信号试剂CaCl₂、NO抑制剂亚甲基蓝(MB)和Ca²⁺通道阻断剂LaCl₃,对紫花苜蓿幼苗光合特征、抗氧化酶活性及过氧化物酶(POD)同工酶图谱进行研究,探讨了渗透胁迫下NO介导的Ca²⁺信号对紫花苜蓿幼苗光合作用及抗氧化能力的影响。结果表明:在渗透胁迫条件下,施加SNP、CaCl₂均能够有效缓解叶片叶绿素a、类胡萝卜素及总叶绿素含量降低,提高叶片净光合速率(P_n)、气孔导度(G_s)及气孔限制值(L_s),而对胞间CO₂浓度(C_i)没有缓解作用。SNP、CaCl₂及SNP+CaCl₂处理提高了幼苗叶片中抗氧化酶活性和脯氨酸含量,降低了丙二醛(MDA)含量。其中共处理时效果最为显著,第4天 SOD、POD、CAT活性较PEG处理升高了39.29%、30.41%和56.24%,脯氨酸含量增加了45.59%,MDA含量降低了45.59%。POD同工酶图谱在第4天时酶谱带数最多,POD活性最强,且SNP+CaCl₂共处理下出现新酶带。而添加外源NO的同时添加Ca²⁺通道阻断剂LaCl₃,紫花苜蓿幼苗光合速率、抗氧化酶活性及脯氨酸含量均降低,丙二醛含量增加,添加Ca²⁺信号的同时施加NO抑制剂MB也具有相同的作用,说明Ca²⁺信号参与NO信号转导过程并相互作用共同调节渗透胁迫下紫花苜蓿幼苗的生理应答响应。     

The evolution and application of enzymes in the animal feed industry: the role of data interpretation

ABSTRACT

1. Enzymes have been used commercially for nearly 40 years and save significant costs through sparing of expensive nutrients but the mechanism by which this is achieved is still debated.

2. The research focused on non-starch polysaccharidase (NSPase) enzymes is used as an example of where greater progress could have been made if the details of the work had been described more fully and the analysis of the data generated had been broader in scope and more critical.

3. Lack of standardisation of the details presented in the materials and methods has been identified as a significant barrier to meaningful retrospective analysis and thus limits advances in the understanding of the mode of action of these enzymes.

4. The identity of the enzyme employed and its activity is often lacking, and more importantly the purity is rarely disclosed. Contaminant activities which are neither listed nor assayed could play a significant role in the responses observed.

5. The dose optimum of most enzymes is often considerably higher than that employed in most studies. Thus studies claiming synergy between two ‘activities’ should ensure that the response is not related to each enzyme simply augmenting the dose of just one activity in the finished feed. This is a common problem, and coupled with the lack of factorial experiments to justify the presence of each enzyme in a multi-enzyme product, it is not surprising that there is still debate as to whether single or multi-enzymes are best suited poultry rations.

6. The three proposed mechanisms for NSPases (viscosity, cell wall and prebiotic) are discussed, and along with their strengths and weaknesses it is suggested that a re-evaluation of each is needed. Viscosity may have to be re-evaluated as being a function not only of the cereal being fed, but of the age of the animal as well. The cell wall theory as described is poorly modelled in vitro and hence the validity of these data is questioned. The prebiotic theory may need significant modification as it appears that the quantities of oligomers produced are insufficient to generate the additional volatile fatty acids (VFA)’s reported. It is likely that all three mechanisms play a role in the responses observed, but the prebiotic mechanism probably plays by far the most important part in low viscosity diets.

7. Future research would be improved if it considered all potential mechanisms when designing a trial. Significant failings are apparent as a result of adherence to tenets in explanation of the results. Most importantly, it should be emphasised that a hypothesis is there to be tested, not defended.     

Noninferiority trial investigating the efficacy of a nonantibiotic intramammary therapy in the treatment of mild‐to‐moderate clinical mastitis in dairy cows with longer lasting udder diseases

A nonblinded, positively controlled, noninferiority trial was conducted to evaluate the efficacy of an alternative, nonantibiotic therapy with Masti Veyxym^® to reduce ineffective antibiotic usage in the treatment of nonsevere clinical mastitis (CM) in cows with longer lasting udder diseases. The solely intramammary treatment with Masti Veyxym^® (three applications, 12 hr apart) and the combined treatment with Masti Veyxym^® and antibiotics as usual on the farm according to label of the respective product were compared with the reference treatment of solely antibiotic therapy. The matched field study was conducted on eight free‐stall dairy farms located in Eastern Germany. Cases of mild‐to‐moderate CM in cows with longer lasting high somatic cell counts in preceding dairy herd improvement test days and with previous CM cases in current lactation were randomly allocated to one of the three treatment groups. A foremilk sample of the affected quarter was taken before treatment and again approximately 14 days and 21 days after the end of therapy for cyto‐bacteriological examination. Primary outcomes were clinical cure (CC) and no CM recurrence within 60 days after the end of treatment (no R60). Bacteriological cure (BC) and quarter somatic cell count (QSCC) cure were chosen as secondary outcomes although low probabilities of BC and QSCC cure for selected cows were expected. The study resulted in the following findings: the pathogens mostly cultured from pretreatment samples were Streptococcus uberis, followed by Staphylococcus aureus and coagulase‐negative staphylococci. There were no significant differences between the two test treatments in comparison with the reference treatment regarding all outcome variables. The sole therapy with Masti Veyxym^® resulted in a numerically lower likelihood of BC without significant differences to the reference treatment. The combined therapy group showed a numerically higher nonrecurrence rate than the two other treatment groups and noninferiority compared to the reference treatment was proven. Having regard to the selection criteria of cows in this study, the findings indicated that sole treatment with Masti Veyxym^® in nonsevere CM cases may constitute an alternative therapy to reduce antibiotics. However, noninferiority evaluations were mostly inconclusive. Further investigations with a larger sample size are required to confirm the results and to make a clear statement on noninferiority.     

Organic zinc and copper supplementation on antioxidant protective mechanism and their correlation with sperm functional characteristics in goats

Trace minerals feeding had significant effects on sperm production and fertility with better absorption and proper utilization within the body for optimum reproductive function. Several studies have shown that more influenced trace elements in the diets of animals are copper (Cu) and zinc (Zn). Bucks showing deficiency of this mineral might affect the quality of semen production which in turn would affect the fertility. This experiment was thus designed to test the effects of organic Cu and Zn supplementation on antioxidants enzyme activities and sperm functional attributes in fresh semen of bucks. Forty bucks (n = 40, Aged 5 months) were assigned to ten groups of four animals in each group, supplemented (for a period of 8 months) with different levels of organic Zn: 20 mg (T2), 40 mg (T3) and 60 mg (T4), organic Cu: 12.5 mg (T5), 25 mg (T6), 37.5 mg (T7) and combined organic Zn and Cu: 20 + 12.5 mg (T8), 40 + 25 mg (T9), 60 + 37.5 mg (T10), respectively, per kg dry matter and no additional mineral diet (control; T1). One hundred and sixty semen samples were collected through electro‐ejaculator and analysed for sperm quantity, quality, acrosome intactness and plasma membrane integrity and correlated with the catalase, superoxide dismutase, glutathione peroxidase and glutathione reductase enzyme activities in seminal plasma. The results indicated organic Cu and zinc supplemented bucks produced more sperm cells, had higher sperm concentrations, maintained higher (p < .01) sperm livability, plasma membrane and acrosome integrities, more motility and velocity. The increased antioxidant enzyme activities, reduced oxidative stress and lowered lipid peroxidation were positively correlated (p < .05) with the sperm functional attributes. In conclusion, organic Cu and Zn supplement to male goats showed protective roles against oxidative damage and maintained better fresh semen characteristics.     

Copper‐Induced Oxidative Stress and Responses of the Antioxidant System in Roots of Medicago sativa

The effects of various copper (Cu) concentrations on the antioxidative system in the roots of Medicago sativa were explored. The results indicated that the Cu content of the roots reached a value of 854 μg g^?1 DW at 10 μm Cu and a value of 4415 μg g^?1 DW at 100 μm Cu, suggesting that M. sativa has better ability to tolerate and accumulate Cu than other Cu‐bioaccumulators, and is a potential plant for phytoremediation. Treatment with Cu resulted in a significant increment in the levels of H₂O₂, O₂˙^? and OH˙. The reduced form of ascorbate and glutathione reached a peak at 30 μm Cu, and was followed by a sharp depletion to a lower level than that of the control. In contrast, the levels of the oxidised forms of ascorbate and glutathione showed a progressive increment with increasing Cu concentrations, suggesting that the antioxidant system was unable to cope with Cu stress at higher Cu levels. Under the Cu concentrations tested, the activity of catalase (CAT, EC 1.11.1.6), ascorbate peroxidase (APX, EC 1.11.1.11) and glutathione reductase (GR, EC 1.6.4.2) increased at lower Cu concentrations, and then decreased, reaching a maximum at 30 μm of Cu for APX and GR, at 10 μm for CAT, whereas the activities of guaiacol peroxidase (POD, EC 1.11.1.7) were gradually increased with increasing Cu concentrations. PAGE analysis of superoxide dismutase (SOD, EC 1.1.5.1.1) revealed that one band is a Mn‐SOD and five bands are identified as Cu, Zn‐SOD, whereas Fe‐SOD isoforms were not found in the roots of alfalfa. Cu at 10–100 μm increased the intensity of constitutive isozymes of CAT, APX and POD, whereas it decreased the intensity of isozymes of glucose‐6‐phosphate dehydrogenase (G6PDH, EC 1.1.1.49) significantly. The activities of lipoxygenases (LOX, EC 1.13.11.12) were gradually augmented with increasing Cu concentrations, demonstrating that LOXs are probably involved in production of lipid hydroperoxides and superoxide anion. There was a continuous and pronounced enhancement in the activity of esterase (EST, EC 3.1.1.1) in roots treated with 10–30 Cu μm , whereas EST activity in roots exposed to above 30 μm Cu declined, suggesting that EST plays a protective role under lower Cu concentrations stress.     

紫斑牡丹种子浸提液对植物种子萌发和幼苗生长的影响

以紫斑牡丹种子为研究对象,研究不同质量浓度(0、0.025、0.050、0.075、0.100 g·mL-1)的种皮和胚乳水浸提液对白菜、小麦和绿豆种子萌发、幼苗生长和幼苗的抗氧化酶活性的影响,探究紫斑牡丹种子内源抑制物质的活性,为进一步研究紫斑牡丹种子休眠机理提供理论依据。结果表明:紫斑牡丹种皮和胚乳中均含有抑制受体植物种子萌发和幼苗生长的物质,且同一质量浓度下,胚乳浸提液的抑制作用强于种皮浸提液;种皮和胚乳浸提液对白菜幼苗生长的抑制作用高于小麦和绿豆;种皮和胚乳浸提液对3种受体植物幼苗根长的抑制作用大于苗高和鲜质量;种皮和胚乳浸提液对3种受体植物幼苗的SOD、CAT和POD活性均产生了一定的影响。其中,随着种皮浸提液质量浓度的提高,受体植物幼苗SOD、POD活性先上升后下降,而CAT活性持续下降;加入胚乳浸提液后,受体植物幼苗的CAT、POD活性与对照相比显著下降,SOD活性则依然随着胚乳浸提液质量浓度的升高而表现出先上升后下降的趋势。由结果可知,紫斑牡丹种皮和胚乳浸提液中均含有能抑制受体植物种子萌发和幼苗生长的物质,并且能够影响受体幼苗的抗氧化酶活性,但是紫斑牡丹种皮和胚乳内源抑制物质的活性存在一定的差异。     

氟节胺抑制枇杷嫁接成苗砧木萌蘖相关机理初探

为寻找更长效、无害的化学控蘖剂以减轻田间管理劳动强度,以枇杷(Eriobotrya japonica)嫁接成苗为材料,喷施有效成分浓度为0.050%,0.071%和0.125%的氟节胺,于不同时期取样观察控蘖效果,并尝试探究相关机理。结果表明,氟节胺减少了叶片单位面积光能的吸收量、抑制了光化学转换,但叶片的热耗散量并无明显变化。进一步测定植株氧化还原状态发现,氟节胺能够快速促进细胞内超氧阴离子(O₂^.-)积累并延缓或抑制H₂O₂的转化,从而诱发氧化胁迫。由此可见,氟节胺可能通过抑制叶片光化学转换和热耗散来破坏新生萌蘖的光合性能,进而抑制其萌发生长,而成熟叶片则通过提升自身抗氧化系统活性耐受氟节胺造成的轻度胁迫。在本试验中,各浓度的氟节胺对控制砧木萌蘖有不同程度的效果,其中浓度为0.125%的氟节胺控蘖效果相对最佳,且基本不会对砧木和嫁接成苗造成持续伤害,其作为化学控蘖剂有一定的应用潜力。     

根腐病菌侵染对黄芪苯丙烷途径关键酶活性的影响

以蒙古黄芪-根腐病菌（Fusarium solani和F. acuminatum）为互作体系,测定苯丙烷途径关键酶苯丙氨酸解氨酶(PAL)、肉桂酸-4-羟化酶(C4H)及4-香豆酸-辅酶A连接酶(4CL)活性的变化,以期获得黄芪抗根腐病菌侵染机制的信息,为抗病育种工作提供参考。试验采用幼苗浸根法接种病菌,分光光度法测定3种酶的动态变化。结果表明,F. solani侵染黄芪后,PAL和C4H活性与对照相比明显升高,并在7、21天出现2个酶活高峰;4CL活性则随着时间的延长持续上升,7、21天时也显著高于对照。F. acuminatum侵染黄芪后,PAL、C4H和4CL的活性均在21天显著高于对照。可见2种病原菌在侵染黄芪过程中,苯丙烷途径的关键酶被不同程度地激活,但2种病菌激活各酶的时间和规律有所差异,表明黄芪对这2种病菌的抗性机制明显不同。     

Soil enzyme activities during the 2011 Texas record drought/heat wave and implications to biogeochemical cycling and organic matter dynamics

Extreme droughts and heat waves due to climate change may have permanent consequences on soil quality and functioning in agroecosystems. During November 2010 to August 2011, the Southern High Plains (SHP) region of Texas, U.S., a large cotton producing area, received only 39.6 mm of precipitation (vs. the historical avg. of 373 mm) and experienced the hottest summer since record keeping began in 1911. Several enzyme activities (EAs) important in biogeochemical cycling were evaluated in two soils (a loam and a sandy loam at 0–10 cm) with a management history of monoculture (continuous cotton) or rotation (cotton and sorghum or millet). Samplings occurred under the most extreme drought and heat conditions (July 2011), after precipitation resulted in a reduction in a drought severity index (March 2012), and 12 months after the initial sampling (July 2012; loam only). Eight out of ten EAs, were significantly higher in July 2011 compared to March 2012 for some combinations of soil type and management history. Among these eight EAs, enzymes key to C (β-glucosidase, β-glucosaminidase) and P cycling (phosphodiesterase, acid and alkaline phosphatases) were significantly higher (19–79%) in July 2011 than in March 2012 for both management histories regardless of the soil type (P > 0.05). When comparing all sampling times, the activities of alkaline phosphatase, aspartase and urease (rotation only) showed this trend: July 2011 > March 2012 > July 2012. Activities of phosphodiesterase, acid phosphatase, α-galactosidase, β-glucosidase and β-glucosaminidase were higher in July 2011 than July 2012 in at least one of the two management histories. Total C was reduced significantly from July 2011 to March 2012 in the rotation for both soils. Only the activities of arylsulfatase (avg. 36%) and asparaginase showed an increase from July 2011 to March 2012 for both soil types, which may indicate they have a different origin/location than the other enzymes. EAs continued to be a fingerprint of the soil management history (i.e., higher EAs in the rotation than in monoculture) during the drought/heat wave. This study provided some of the first evidence of the adverse effects of a natural, extreme drought and heat wave on soil quality in agroecosystems as indicated by EAs involved in biogeochemical cycling.