Antibacterial Mechanism of Chitosan–Gentamicin and Its Effect on the Intestinal Flora of Litopenaeus vannamei Infected with Vibrio parahaemolyticus

To explore the application of chitosan–gentamicin conjugate (CS-GT) in inhibiting Vibrio parahaemolyticus (V. parahaemolyticus), which is an important pathogen in aquatic animals worldwide, the antimicrobial activity of CS-GT and the effects of a CS-GT dose on the intestine histopathology and intestinal flora of V. parahaemolyticus-infected shrimps were explored. The results showed that CS-GT possessed broad-spectrum antibacterial activity, with minimum inhibitory concentration (MIC), minimum bactericidal concentration (MBC) and half inhibitory concentration (IC₅₀) of 20.00 ± 0.01, 75.00 ± 0.02 and 18.72 ± 3.17 μg/mL for V. parahaemolyticus, respectively. Further scanning electron microscope and cell membrane damage analyses displayed that the electrostatic interaction of CS-GT with cell membrane strengthened after CS grafted GT, resulting in leakage of nucleic acid and electrolytes of V. parahaemolyticus. On the other hand, histopathology investigation indicated that high (100 mg/kg) and medium (50 mg/kg) doses of CS-GT could alleviate the injury of a shrimp’s intestine caused by V. parahaemolyticus. Further 16S rRNA gene sequencing analysis found high and medium dose of CS-GT could effectively inhabit V. parahaemolyticus invasion and reduce intestinal dysfunction. In conclusion, CS-GT possesses good antibacterial activity and could protect shrimps from pathogenic bacteria infection.     

诱导剂对番木瓜环斑病毒病的防治效果

为了探讨水杨酸、茉莉酸甲酯和壳聚糖对番木瓜环斑病毒病的病情指数和防治效果的影响,在番木瓜上喷施不同浓度的诱导剂,然后在叶腋处接种番木瓜环斑病毒,观察和记录番木瓜的病情指数,计算其防治效果.结果表明,诱导剂提高了番木瓜对环斑病毒病的抗性,其中水杨酸的效果优于茉莉酸甲酯和壳聚糖.水杨酸、茉莉酸甲酯和壳聚糖的最佳施用浓度分别为50 mg/L、0.05 mmol/L和1％,最佳防治效果则分别为77.19％、53.11％和22.95％.水杨酸、茉莉酸甲酯、壳聚糖与对照之间的防治效果差异极显著,3种诱导剂在最佳浓度下防治效果差异极显著.50 mg/L水杨酸对番木瓜环斑病毒病防治效果最佳,在生产上施用具有一定的经济效益.     

Therapeutic Intervention with Dietary Chitosan Nanoparticles Alleviates Fish Pathological and Molecular Systemic Inflammatory Responses against Infections

Marine bio-sourced chitosan nanoparticles (CSNP) are antimicrobial and immunomodulatory agents beneficial for fish medicine. Herein, dietary CSNP was investigated for the amelioration of the systemic inflammatory responses of an induced fish model. One hundred and forty-four rainbow trout were assigned to one pathogen-free and non-supplemented group (negative control), and three challenged groups: non-supplemented (positive control), CSNP-preventive, and CSNP-therapeutic. After a feeding experiment extended for 21 days, the organosomatic indices (OSI) and molecular aspects were assessed. After a challenge experiment extended for further 28 days, CSNP-therapeutic intervention was assessed on fish survival and systemic inflammatory responses on pathology, histo-morphology, and molecular aspects. With CSNP administration, OSI nonsignificantly decreased and the relative expression of targeted inflammatory-mediator genes was significantly increased. The CSNP-therapeutic fish showed an RPS of 80% as compared to the positive control group, and CSNP-therapeutic administration retained the highest gene expression augmentation up to 28 days after the challenge. Notably, the splenic reticulin fibers framework of the CSNP-therapeutic group retained the highest integrity among the groups during the infection. After recovery, reticulin fibers density in the CSNP-therapeutic samples was significantly higher than in the negative control group, which indicates high innate immunity. Thus, CSNP showed promising biotherapeutic features enhancing fish resistance against infections.     

不同分子量壳聚糖对白鲢鱼丸货架期品质的影响

将壳聚糖应用于白鲢鱼糜制品的保鲜,通过时白鲢鱼丸货架期的pH值、挥发性盐基总氮、TBA值、过氧化值以及菌落总数的测定,研究了壳聚糖对鱼丸货架期品质的影响.鱼丸中添加1%的壳聚糖成分即可有效抑制其脂质氧化与微生物的生长.壳聚糖对鱼丸的保鲜功能取决于其分子量大小,与高分子量的壳聚糖相比,低分子量的壳聚糖具有更强的抗氧化能力,且30万分子量与1万分子量壳聚糖的混合物具有更强的抗菌能力.     

干旱胁迫下壳寡糖对玉米叶片叶绿素含量、含水量及保护酶活性的影响

以玉米为研究对象,设置壳寡糖和清水（对照）2个处理,研究了在干旱胁迫下外施壳寡糖对玉米叶片叶绿素含量、含水量及叶片保护酶（SOD、POD、CAT）活性的影响。结果表明,在干旱胁迫下,壳寡糖处理和对照均使玉米叶片中叶绿素含量表现为先上升后下降的趋势,但壳寡糖处理叶绿素含量由断水后第7天的30.7 mg/g 提高到断水后第15天的32.4 mg/g ,而对照则由断水后第7天的27.4 mg/g下降至21.4 mg/g;壳寡糖处理和对照的叶片相对含水量在断水后均呈下降趋势,但壳寡糖处理在断水第15天较对照增加16.2％,此时对照表现为萎焉（叶片含水量52.4％）,壳寡糖处理则生长正常,外施壳寡糖具有减缓 SOD、POD、CAT活性下降的作用。综合分析认为,外施壳聚糖可提高玉米对干旱的抵抗能力。     

采前壳聚糖对厚皮甜瓜果实潜伏侵染及其采后主要病害的控制

采用1 g/L的壳聚糖分别在"银帝"甜瓜开花前1周、幼果期、果实膨大期、网纹形成期对植株进行1、2、3、4次喷洒.结果表明:采前壳聚糖处理可有效降低甜瓜果实采后病害的真菌潜伏侵染率、Alternaria alternata和Fusariumspp.的潜伏侵染率.随着处理次数的增加,潜伏侵染率依次降低,其中以4次处理效果最好.此外,采前壳聚糖3次和4次处理还可有效抑制采后果实损伤接种A.alternata和F.semitectum病斑面积的扩大.     

鱼肉酶解物及壳聚糖对鲤鱼涂膜保鲜效果的研究

以感官评分、挥发性盐基氮、菌落总数、K值为指标,研究比较了鱼肉酶解物和壳聚糖为原料的涂膜液对鲤(Cyprinus carpio)(4℃)冷藏过程中品质变化的影响。结果显示:贮藏期间,对照组的感官评分显著低于各涂膜组(P<0.05)。贮藏前8 d,涂膜组能够显著抑制K值的升高(P<0.05)。鱼肉酶解物和壳聚糖涂膜组均能够显著抑制细菌的生长,而贮藏2～6 d内鱼肉酶解物涂膜鲤鱼的菌落总数显著低于壳聚糖涂膜组(P<0.05)。贮藏后期,鱼肉酶解物组能延缓TVB-N值的升高,壳聚糖涂膜组能够显著抑制TVB-N值的升高(P<0.05)。鱼肉酶解物可作为一种新的可食性涂膜材料,用于延长鲤鱼的贮藏期。     

壳聚糖对Cd2+和Pb2+的吸附作用

为开发新型吸附剂,降低重金属对水体的污染,研究了壳聚糖对Cd²⁺和Pb²⁺的吸附条件,探讨了pH、温度、反应时间、壳聚糖添加量和金属离子初始浓度等因素对壳聚糖吸附性能的影响。结果表明,pH 7~8和pH 5~6条件下壳聚糖对Cd²⁺和Pb²⁺的吸附能力最强;低温有利于壳聚糖的吸附;在8 h时壳聚糖对Cd²⁺的吸附容量达到最大,而对Pb²⁺的吸附在实验时间内是随着吸附时间的延长而增大;随着壳聚糖添加量的增加,其对Cd²⁺和Pb²⁺的吸附能力也增强;初始金属离子浓度的变化对Cd²⁺的影响不大,而在高的金属浓度下对Pb²⁺的吸附率显著降低。壳聚糖对Cd²⁺、Pb²⁺的吸附动力学和热力学分别符合Lagergren方程二级吸附模型和Langmuir吸附方程。研究表明,壳聚糖对不同金属离子的吸附能力不同,在单一金属溶液中,壳聚糖对Cd²⁺的吸附能力要强于对Pb²⁺的吸附能力。系统地研究了壳聚糖对Cd²⁺和Pb²⁺的吸附条件及性能,为壳聚糖作为重金属吸附剂的应用提供理论依据。     

胰蛋白酶对壳聚糖的降解研究

用粘度法和吸光度法，研究胰蛋白酶非专一性降解壳聚糖过程中温度、PH值、反应时间、酶浓度、底物浓度。金属离子对胰蛋白酶降解壳聚糖反应的影响，确定了以壳聚糖为底物的胰蛋白酶的一些催化特性：最适温度为30℃，最适PH值为5.0,10-180min内酶反应速度恒定，酶浓度在0.1-0.5g/L的范围内，酶反应速度与酶浓度呈线性关系，米氏常数Km为9.54g/L。