A Lymphoid Organ Specific Anti-Lipopolysaccharide Factor from Litopenaeus vannamei Exhibits Strong Antimicrobial Activities

Different shrimp species are known to possess apparent distinct resistance to different pathogens in aquaculture. However, the molecular mechanism underlying this finding still remains unknown. One kind of important antimicrobial peptides, anti-lipopolysaccharide factors (ALF), exhibit broad-spectrum antimicrobial activities. Here, we reported a newly identified ALF from the shrimp Litopenaeus vannamei and compared the immune function with its counterpart in the shrimp Fenneropenaeus chinensis. The ALF, designated as LvALF8, was specifically expressed in the lymphoid organ of L. vannamei. The expression level of LvALF8 was apparently changed after white spot syndrome virus (WSSV) or Vibrio parahaemolyticus challenges. The synthetic LBD peptide of LvALF8 (LvALF8-LBD) showed strong antibacterial activities against most tested Gram-negative and Gram-positive bacteria. LvALF8-LBD could also inhibit the in vivo propagation of WSSV similar as FcALF8-LBD, the LBD of LvALF8 counterpart in F. chinensis. However, LvALF8-LBD and FcALF8-LBD exhibited apparently different antibacterial activity against V. parahaemolyticus, the main pathogen causing acute hepatopancreatic necrosis disease (AHPND) of affected shrimp. A structural analysis showed that the positive net charge and amphipathicity characteristics of LvALF8-LBD peptide were speculated as two important components for its enhanced antimicrobial activity compared to those of FcALF8-LBD. These new findings may not only provide some evidence to explain the distinct disease resistance among different shrimp species, but also lay out new research ground for the testing and development of LBD-originated antimicrobial peptides to control of shrimp diseases.     

基于注意力机制及多尺度特征融合的番茄叶片缺素图像分类方法

针对番茄早期缺素性状不明显及各生长期特征差异较大所导致的特征区域尺寸不一致、难提取、难辩别等问题，提出了一种基于注意力机制及多尺度特征融合卷积神经网络的番茄叶片缺素图像分类方法（Multi-Scale Feature Fusion Convolutional Neural Networks Based On Atte ntion Mechanism，MSFF-AM-CNNs）。首先根据番茄叶片缺素特点提出了多尺度特征融合结构（Multi-Scale Feature Fusion Module，MSFF Module）；其次在DenseNet基础上，结合浅层网络主要提取纹理、细节特征，深层网络主要提取轮廓、形状特征的特点分别提出具有针对性的特征提取方法，通过不同形式引入注意力机制及多尺度特征融合结构，使全局多尺度信息融合多个特征通道、选择性地强调信息特征并达到对特征精准定位的功能；同时引入Focal Loss函数以减少易分类样本的权重。试验结果表明，MSFF-AM-CNNs的平均召回率、平均F1得分、平均准确率较原模型DenseNet-121均大幅提升，其中缺氮和缺钾叶片的准确率分别提高了8.06和6.14个百分点，召回率分别提高了6.31和5.00个百分点，F1得分分别提高了7.25和5.55个百分点，平均识别准确率可达95.92%，具有较高的识别准确率及广泛的适用性，能够满足番茄叶片缺素图像的高精度分类需求，可为植物叶片缺素识别提供参考。     

蛋鸡脂肪肝综合征模型的建立

试验旨在建立蛋鸡脂肪肝综合征病理模型。选取5日龄的海兰褐蛋鸡公雏280只，随机分为对照组（基础日粮组）和3个模型组（A、B、C组），试验第1~10天给模型组饲喂不同配比的高脂饲料，第11~20天饲喂常规基础日粮，每天观察并记录试验鸡精神状态、外观体征、饮水量和食欲情况，试验第0、10、20天从各组随机抽取15只鸡进行翅静脉采血并剖取肝脏和腹脂，测定肝脏系数、肝脂率、腹脂率，血清总胆固醇（TC）、甘油三酯（TG）、谷丙转氨酶（ALT）和谷草转氨酶（AST）水平。结果显示，试验第10天，3个模型组剖检时可见腹腔和肠系膜有大量的脂肪沉积，肝脏系数、肝脂率和腹脂率，以及血清中总胆固醇、甘油三酯、谷丙转氨酶和谷草转氨酶指标均符合鸡脂肪肝综合征模型的诊断标准；试验第20天，模型A、B组谷丙转氨酶和谷草转氨酶水平有所恢复，其他指标仍高于对照组，而模型C组的临床症状和各项检测指标仍符合鸡脂肪肝综合征模型的诊断标准。因此采用连续饲喂高脂饲料C（74.5%基础日粮、6%胆固醇、14%猪油、5%蔗糖、0.5%丙基硫氧嘧啶）可成功建立蛋鸡脂肪肝综合征模型。     

WSSV envelope protein VP51B links structural protein complexes and may mediate virus infection

White spot syndrome virus (WSSV), an enveloped double‐stranded DNA virus, is the causative agent of a disease that has led to severe mortalities of cultured shrimps in Taiwan and many other countries. In the previous study, Penaeus monodon chitin‐binding protein (CBP) and glucose transporter 1 (Glut1), two cell membrane proteins, were found to at least interact with other 10 WSSV envelope proteins including VP51B. These envelope proteins might form a protein complex. According to the known information, VP51B was used to identify its role in the protein complex. Western blotting of the intact viral particles and fractionation of the viral components confirmed that VP51B is one of WSSV envelope proteins. In this study, the protein–protein interaction between VP51B and other WSSV envelope proteins was identified by far‐western blot experiment and VP51B was found to interact with VP24, VP31, VP32, VP39B and VP41A. Furthermore, the in vivo neutralization experiment using recombinant VP51B plus with VP39B showed the best inhibition. These data indicate that VP51B participates in the WSSV protein complex and plays an important role in WSSV infection.     

花前氮亏缺对水稻叶片氮代谢酶活性的影响

以籼型杂交稻中浙优1号和籼粳型杂交稻甬优12为材料,试验分析了花前不同时期氮亏缺处理对水稻叶片氮代谢酶活性的影响。结果表明,花前氮亏缺导致植株上3叶氮浓度、NR、GS、GOGAT、GOT和GPT酶活性大幅下降,GDH活性显著增长,且各叶位对花前氮亏缺敏感度总体上表现为剑叶倒2叶倒3叶;破口期亏氮对籼型杂交稻中浙优1号上3叶氮同化酶活性的影响远小于减数分裂期处理,与之相比,籼粳杂交稻甬优12对破口期亏氮胁迫仍较敏感,表明中浙优1号植株氮代谢酶的亏氮敏感性由减数分裂期至破口期逐步下降,对土壤速效氮的需求同步降低,甬优12则对土壤供氮存在更高需求。     

In vitro and in vivo selection of probiotic purple nonsulphur bacteria with an ability to inhibit shrimp pathogens: acute hepatopancreatic necrosis disease‐causing Vibrio parahaemolyticus and other vibrios

Shrimp cultivation has been faced with huge losses in productivity caused by infectious shrimp pathogenic vibrios, especially Vibrio parahaemolyticus that causes acute hepatopancreatic necrosis disease (AHPND). Hence, purple nonsulphur bacteria (PNSB) were isolated from shrimp ponds for investigating their abilities to control shrimp pathogenic Vibrio spp. and their use as probiotics for sustainable shrimp cultivation. Based on their probiotic properties, strains S3W10 and SS15 were selected because of their strong abilities to produce amylase, gelatinase and vitamin B12. However, only three PNSB strains (SS15, TKW17 and STW181) strongly inhibited V. harveyi_KSAAHRC and V. vulnificus_KSAAHRC including V. parahaemolyticusAHPND strains by secreting antivibrio compounds. Four selected PNSB also grew in the presence of pancreatic enzymes, and they were identified as Rhodobacter sphaeroides for strains S3W10, SS15 and TKW17 and Afifella marina for strain STW181. The effects of a mixed culture were also investigated as follows: T1 (S3W10 + SS15), T2 (S3W10 + TKW17) and T3 (S3W10 + STW181) on postlarval white shrimp (Litopenaeus vannamei) for 60 days by comparison with a control. All three probiotic PNSB sets significantly improved the digestive enzyme activities and shrimp growth with their proliferation in shrimp gastrointestinal tract although the shrimp survival was not significantly different. They also significantly reduced the cumulative mortality of shrimp exposed to a virulent AHPND strain (V. parahaemolyticusSR2). This is the first to conclude that selected probiotic PNSB strains have great potential to be used for shrimp cultivation to control vibrios including AHPND strains.     

Agronomic traits at the seedling stage,yield, and fiber quality in two cotton (Gossypium hirsutum L.) cultivars in response to phosphorus deficiency

ABSTRACT

Cotton is critical for phosphorus demands and very sensitive for its deficiency. However, identifying the effect of low-phosphorus tolerance on cotton growth, yield, and fiber quality by reducing phosphorus consumption. This may help to develop phosphorus-tolerant high-yielding cotton cultivars. In a two-year repeated (2015 and 2016) hydroponic experiment (using 0.01 and 1 mM KH₂PO₄), two cotton cultivars with phosphorus sensitivity (Lu 54; a low-phosphorus sensitive and Yuzaomian 9110; a low-phosphorus tolerant) were screened on the base of agronomic traits and physiological indices through correlation analysis, cluster analysis and principal component analysis from 16 cotton cultivars. Low phosphorus nutrition reduced the plant height, leaf number, leaf area, phosphorus accumulation and biomass in various organs of seedlings. The deficiency negatively affected the morphogenesis of seedlings, as well as yield and fiber quality. Further, these screened cultivars were tested in a pot experiment with 0, 50, 100, 150, 200 kg P₂O₅ ha^?1 during 2016 and 2017. It was found to have a significant (P< 0.05) difference in boll number, lint yield, fiber strength, and micronaire at the harvest. Furthermore, after collectively analyzed the characteristics of Lu 54 and Yuzaomian 9110, there were six key indices that could improve the low phosphorus tolerance of cotton cultivars. These were root phosphorus accumulation, stem phosphorus accumulation percentage, leaf and total biomass of seedlings, seed cotton weight per boll and fiber length.     

Effect of dietary calcium or phosphorus deficiency on bone development and related calcium or phosphorus metabolic utilization parameters of broilers from 22 to 42 days of age

This experiment was conducted to investigate the effect of dietary calcium (Ca) or phosphorus (P) deficiency on bone development and related Ca or P metabolic utilization parameters of broilers from 22 to 42 days of age based on our previous study, which indicated that dietary Ca or P deficiency impaired the bone development by regulating related Ca or P metabolic utilization parameters of broilers from 1 to 21 days of age. A total of 504 one-day-old Arbor Acres male broilers were randomly assigned to 1 of 4 treatments with 7 replicates in a completely randomized design, and fed the normal control and Ca- or P-deficient diets from 1 to 21 days of age. At 22 days of age, the broilers were further fed the normal control diet (0.90% Ca+0.35% non-phytate P (NPP)), the P-deficient diet (0.90% Ca+0.18% NPP), the Ca-deficient diet (0.30% Ca+0.35% NPP) or the Ca and P-deficient diet (0.30% Ca+0.18% NPP), respectively. The results showed that dietary Ca or P deficiency decreased (P<0.05) tibia bone mineral density (BMD), bone breaking strength (BBS), ash content, tibia ash Ca content and serum P content on days 28 and 42, but increased (P<0.05) tibia alkaline phosphatase (ALP) activity of broilers on day 42 compared with the control group. Furthermore, the broilers fed the P-deficient diet had the lowest (P<0.05) tibia BMD, BBS, ash content, serum P content and the highest (P<0.05) serum Ca content on day 28 compared with those fed the Ca-deficient or Ca and P-deficient diets. The results from the present study indicated that the bone development and related Ca or P metabolic utilization parameters of broilers were the most sensitive to dietary P deficiency, followed by dietary Ca deficiency or Ca and P-deficiency; dietary Ca or P deficiency impaired the bone development possibly by regulating serum Ca and P contents as well as tibia Ca content and ALP activity of broilers from 22 to 42 days of age.     

Role of cytokeratin 8 on changes of intestinal epithelial permeability induced by corticotropin-releasing factor

AIM: To investigate the role of cytokeratin 8 (CK8) on the change of intercellular permeability of intestinal epithelial cells induced by corticotropin-releasing factor (CRF). METHODS: The expression levels of CRF receptor 1 (CRFR1) and CRFR2 on human colon adenocarcinoma HT29 cell surface were determined by immunofluorescence staining. After treatment with 100 nmol/L CRF for 72 h, the translocation of FITC-labelled dextran was measured in a Transwell chamber. The structural changes of tight junctions were observed under transmission electron microscope. The expression levels of CK8, and tight junction proteins ZO-1 and occludin were determined by Western blot. The activity of protein kinase C (PKC) was detected by ELISA. Furthermore, the effects of CRF on intestinal epithelial permeability were examined in CK8-silencing HT29 cells, which were constructed by infection with sh-CK8 lentivirus. RESULTS: CRF treatment increased the permeability of FITC-labelled dextran (P<0.05), caused the opening of tight junctions, and induced increased fluorescence intensity of CK8. The expression levels of occludin and ZO-1 were down-regulated (P<0.05). PKC activity was decreased at 1 h after CRF treatment (P<0.05). CRF-induced increase in the permeability and down-regulation of occludin were not blocked by CK8 silencing. Nevertheless,CK8 silencing blocked the effects of CRF regarding the decrease in the expression levels of ZO-1 and the increase in PKC activity (P<0.05). CONCLUSION: CK8 may be involved in CRF-induced increase in intestinal epithelial permeability by inhibiting the activity of PKC, and there may be other signaling pathways involved.