The future of genetic engineering to provide essential dietary nutrients and improve growth performance in aquaculture: Advantages and challenges

Advancements in gene technology in recent years have been driving the aquaculture industry forward. Improvements in growth performance, feed efficiency, and omega‐3 content are goals of the industry that could capitalize on applications of genetic engineering. One of the major challenges in the industry is to reduce the use of fish meal and oil, to improve the environmental and economic sustainability of aquaculture. The recent development of genetically engineered feed ingredients is one potential solution to the looming problem of fish meal and oil dependency. Furthermore, the development of transgenic fish has potential to improve production efficiency and other future desirable characteristics that relate to feed utilization and product quality. New gene technologies are beginning to revolutionize how we produce our food, and in aquaculture, will ultimately reduce pressure on wild fish stocks, help to preserve natural aquatic ecosystems, and improve nutritional profiles of farmed fish for human consumption. The purpose of this review is to provide an update on the current applications of genetic engineering technology to improve aquaculture through nutrition, including the development and use of transgenic feed ingredients, transgenic fish, and ultimately their impacts on nutrition, product quality, and consumers.     

Secondary Stress Responses in Juvenile Brazilian Flounder,Paralichthys orbignyanus,throughout and after Exposure to Sublethal Levels of Ammonia and Nitrite

This study investigated the secondary stress responses of Paralichthys orbignyanus exposed to ammonia and nitrite and after recovery. Fish were exposed to 0.12, 0.28, and 0.57 mg NH₃‐N/L, or 5.72, 10.43, and 15.27 mg NO₂‐N/L for 10 d followed by the same time length for recovery. Ammonia‐ and nitrite‐free water was used as a control treatment. Blood samples were collected after 1, 5, and 10 d of exposure and after recovery. Fish exposed to ammonia presented lower and higher glucose levels after 10 d of exposure and recovery, respectively. Ammonia induced initial and transient ionic disturbances and metabolic alkalosis. Nitrite exposure caused hyperglycemia, increased plasma K⁺ levels, and respiratory alkalosis, whereas metabolic acidosis was observed after recovery. Increased proportion of monocytes and/or granulocytes and reduced number of lymphocytes were demonstrated in fish exposed to 0.28 mg NH₃‐N/L (Day 1) and 10.43 mg NO₂‐N/L (Day 5) and after recovery in the 0.28 and 0.57 mg NH₃‐N/L treatments. Exposure to ammonia decreased the proportion of granulocytes on Day 5. In conclusion, exposure to concentrations at 0.12 mg NH₃‐N/L and 5.72 mg NO₂‐N/L provoked physiological disorders in Brazilian flounder. Nonetheless, fish exposed to 5.72 mg NO₂‐N/L following a 10‐d recovery period showed complete resumption of homeostasis.     

The microbial diversity and structure in peatland forest in Indonesia

The microbial community structure and function under forest in tropical peatlands are poorly understood. In this study, we investigated the microbial community structure and diversity in natural peat swamp forest soil, disturbed peat soil and mineral soil in Central Kalimantan, Indonesia, using 454 pyrosequencing. The results showed that the natural peat soil had the greatest fungal species richness (Chao1), which was significantly (p < .05) larger than that in the other two soils. Community structure of both fungi and bacteria in natural peat soil differed significantly from that in the disturbed peat soil (p = .039 and p = .045, respectively). Ascomycota (40.5%) was the most abundant phylum across the three soils followed by Basidiomycota (18.8%), Zygomycota (<0.1%) and Glomeromycota (<0.1%). The linear discriminant analysis with effect size (LEfSe) showed that Ascomycota (p < .05) and genus Gliocephalotrichum (p < .05) dominated in natural peat soil. Functionally, pathotrophs were more abundant in disturbed peat soil (p < .05). Proteobacteria (43.8%) were the most abundant phylum followed by Acidobacteria (32.6%), Actinobacteria (9.8%), Planctomycetes (1.7%). Methylocystis, Telmatospirillum, Syntrophobacter, Sorangium and Opitutus were the more abundant genera in disturbed peat soil, whereas Nevskia and Schlesneria were more abundant in mineral soil and natural peat soil, respectively. The natural peat forest soil supported a more diverse microbiology; however, the land use of such a soil can change its microbial community structure. The results provide evidence that the disturbance of tropical peat land could lead to the introduction and spread of a large number of fungal diseases     

Development of a core set of KASP markers for assaying genetic diversity in Brassica rapa subsp. chinensis Makino

Single‐nucleotide polymorphisms (SNPs) are rapid, economical and reliable genotyping tools. Non‐heading Chinese cabbage (Brassica rapa L. subsp. chinensis Makino) is now an economically important vegetable crop worldwide. In this study, 1,167 SNPs were evaluated for 7polymorphism among 70 representative non‐heading Chinese cabbage inbred lines using a Kompetitive Allele Specific PCR (KASP) genotyping assay. On the basis of identified polymorphisms and the results of a principal component analysis, we selected 50 core SNPs that were balanced sufficiently to provide adequate information for genetic identification. The core SNPs were used for construction of a neighbour‐joining dendrogram that separated the 70 inbred lines into four main groups and several subgroups corresponding to Caixin, Heiyebaicai, Huangxinwu, Naibaicai, Taitsai, Pak‐choi, and Wutatsai. This categorization was superior to that achieved using a dataset of 479 polymorphic SNPs. To confirm the utility of the core SNP markers in genetic identification, we tested their stability and resolution using 162 commercial hybrid cultivars. The SNPs, which represent a cost‐effective, accurate marker set for germplasm analysis and cultivar identification, are suitable for molecular marker‐assisted breeding in non‐heading Chinese cabbage.     

东北地区水稻纹枯病菌致病性及遗传多样性分析

为明确东北地区水稻纹枯病菌致病性及其群体遗传多样性,将2018年采自东北三省13个水稻主产区的病样分离纯化,共获得176个立枯丝核菌菌株。采用离体叶片接种法测定菌株致病力,利用SRAP分子标记技术分析其遗传多样性。致病力测定结果表明,东北三省各地区菌株致病力存在分化现象,供试的176个菌株中,强致病力菌株占全部菌株23.3%;中等致病力菌株占61.9%;弱致病力菌株占14.8%。地理来源与致病力无明显相关性。根据UPGMA聚类分析,当相似系数为0.63时,176个菌株被划分为6个类群。聚类结果与地理来源具有相关性,但与致病力无明显相关性。利用16对SRAP引物分析176个立枯丝核菌菌株遗传多样性,共得到2 237个扩增条带,多态性频率为82.16%;基因多样度(h)、Shannon信息指数(I)分别为0.2827、0.4150。东北各地区立枯丝核菌群体间遗传距离与地理距离呈一定正相关。群体遗传分化系数Gst=0.3319,基因流Nm=1.0063,说明东北地区立枯丝核菌群体遗传分化度较高且群体间存在基因交流。AMOVA分析结果表明,群体内遗传分化为67.84%,遗传变异主要发生在群体内部。     

Replacement of fish oil in plant‐based diets for Pacific white shrimp,Litopenaeus vannamei,by stearine fish oil and palm oil

Stearine fish oil (SFO) and palm oil (PO) have emerged as promising alternatives for the replacement of fish oil (FO) in aquafeeds. This study evaluated the replacement of FO with alternative oils in practical diets for Litopenaeus vannamei. In a clear brackish water study (14.1 g/L) utilizing shrimp (0.29 ± 0.02 g, initial weight), FO was replaced by SFO at inclusion ratios of 100:0, 75:25, 50:50, 25:75, and 0:100 (FO:SFO) and PO as 90% of FO. After 55 days, no significant differences (p < 0.05) in final weight, growth, or survival of shrimp were observed. A second trial (8 weeks) in low‐salinity water (2.1 g/L) with shrimp (0.92 ± 0.02 g, initial weight) evaluated diets with 100% FO, 100% SFO, 90% PO, 90% soybean oil (SO), or 90% flaxseed oil (FXO) as a replacement for FO and four commercially produced diets with 2% of FO, SO, PO, or FXO. One treatment received half rations of the commercial FO diet, and one treatment was based entirely on natural productivity. Results show that the fatty acid profiles of the tail muscle conformed to the lipids of the feed, and highly unsaturated fatty acids (HUFAs) were preserved. Following 8 weeks of culture, there were no significant differences in production performance.     

动态高压微射流协同谷氨酰胺转氨酶复合修饰对鱼明胶凝胶特性和结构的影响

为了提高鱼明胶的凝胶特性,本试验采用动态高压微射流(DHPM)协同谷氨酰胺转氨酶(TG)交联(DHPM-TG)技术对鱼明胶进行复合修饰。结果表明,鱼明胶的亮度、特性粘度和胶融温度均随着TG含量的增加而增大,凝胶强度和硬度先增大后减小。与TG修饰技术相比,DHPM-TG复合修饰技术可显著提高鱼明胶的特性粘度、胶融温度、凝胶强度以及胶体的硬度和咀嚼性。此外,环境扫描电镜结果表明,DHPM-TG制备的鱼明胶胶体结构更加致密。综上,DHPM-TG复合修饰显著改善了鱼明胶的凝胶特性,为生产可替代哺乳动物明胶的高品质鱼明胶提供了新的理论依据。     

培肥措施对复垦土壤微生物碳氮代谢功能多样性的影响

土壤复垦是矿区生态环境恢复和耕地总量平衡及质量提升的根本要求。本研究依托山西襄垣采煤塌陷区复垦定位试验基地，采用Biolog-ECO方法和荧光定量PCR技术，研究了不施肥（CK）、单施化肥（CF）、单施有机肥（M）和有机无机培肥（MCF）4种培肥措施下复垦4年和8年土壤微生物碳代谢功能多样性及氮代谢功能基因丰度的变化特征。结果表明，随复垦年限增加，单施有机肥较其他处理可显著提高复垦土壤微生物的总碳源利用能力；不同处理复垦土壤微生物碳源相对利用率总体表现为氨基酸类>糖类>聚合物类>羧酸类>双亲化合物类>胺类，其中单施有机肥更大程度上提高了羧酸类、氨基酸类和胺类碳源的利用率；复垦年限和培肥措施没有改变复垦土壤微生物优势度指数，但有机无机配施较其他处理可显著提高香浓指数（H′）和Pielou均匀度指数；不同处理复垦土壤氮转化功能基因丰度总体表现为amoA（AOA）> amoA（AOB）>nisS、nirK> nifH，5种功能基因丰度均为以有机无机培肥处理最高，且随复垦时间增加而增加；复垦土壤有机质含量与nirS、nirK、nifH基因丰度以及AWCD值存在显著相关性，相关系数在0.707～0.807，同时5种氮转化功能基因丰度均与玉米产量存在显著或极显著的相关性，相关系数在0.824～0.949。综上所述，单施有机肥可提高土壤有机质含量，进而增强了复垦土壤碳代谢强度，有机无机培肥则更有利于复垦土壤碳氮代谢功能多样性的提升，并促进作物产量形成。     

广东梅州柑橘黄龙病和病毒病的发生调查及其黄龙病菌原噬菌体多样性

【目的】调查柑橘黄龙病（Huanglongbing,HLB）和病毒病在广东省梅州市柑橘主产区的危害情况,分析该地区柑橘黄龙病菌（CLas）的原噬菌体多样性。【方法】以16S rDNA为模板设计引物,利用实时荧光定量PCR（qPCR)检测梅州市不同抽检地在2017—2019年3年间柑橘黄龙病的发病情况;同时分别采用已报道的柑橘衰退病毒（Citrus tristeza virus,CTV）、柑橘裂皮病毒（Citrus exocortis viroid,CEVd）、柑橘褪绿矮缩病毒（Citrus chlorotic dwarf-associated virus,CCDaV）、柑橘碎叶病毒（Citrus tatter leaf virus,CTLV）和柑橘黄脉病毒（Citrus yellow vein clearing virus,CYVCV）的特异性检测引物,提取样品的RNA,反转后以样品的cDNA为模板,通过普通PCR的方法分析梅州市抽检地区柑橘病毒病的发生情况。此外,以基于2种原噬菌体类型（SC1和SC2）对应的超变异基因区域设计的2对引物（Lap-TJ-F/Lap-TJ-R1和Lap-TJ-F/Lap-TJ-R2）,运用普通PCR方法分析该地区CLas菌株原噬菌体的遗传多样性。【结果】除2018年9月梅州大浦顺兴公司蜜柚基地的抽检样品外,梅州市其他被检地区均发现CLas。总体来说,梅州市的脐橙CLas检出率为55.3%,蜜柚为61.5%,沙田柚为61.7%。其中2017年采集自平原县大拓镇的蜜柚和沙田柚样品CLas检出率为100%,兴宁市白马镇的蜜柚为100%,沙田柚为80%;其他抽检地区的样品CLas检出率在16.7%—83.3%。虽然梅州市部分地区的检出率较高,但其病原菌的含量并不高,大部分处于柑橘黄龙病发病初、中期,属可防可控阶段;此外,梅州市柑橘病毒的总检出率不高,CTV、CTLV和CYVCV为梅州的主要病毒,CEVd和CCDaV没有检测到。CTV、CTLV和CYVCV在脐橙上的检出率依次为78.9%、7.9%和21.1%,蜜柚为15.4%、25.0%和9.6%,沙田柚为6.4%、2.1%和4.3%。基于Lap-TJ-F/Lap-TJ-R1和Lap-TJ-F/Lap-TJ-R2来研究CLas原噬菌体多样性的PCR扩增条带共有4种类型（SC1-1、SC1-2、SC2-1和SC2-2）,经过PCR电泳和测序得出,来自梅州市脐橙和沙田柚的CLas菌株以SC2-1型为主,蜜柚上以SC1-1型为主。【结论】梅州柑橘产区的柑橘黄龙病目前属于可防可控阶段,其病原菌菌株的原噬菌体在不同品种上具有其独特性;由于在抽检时发现有柑橘病毒病的存在,因此在种植过程中需加强种苗监管,同时应采取有效措施对柑橘黄龙病和柑橘木虱（Diaphorina citri）进行绿色防控。