温度与光照强度对浒苔孢子/配子放散和萌发的影响

通过比较不同温度和光照强度对浒苔(Ulva prolifera)孢子/配子放散和萌发的影响,来探究适合浒苔孢子/配子放散和萌发的温度和光照强度。实验结果表明,适宜浒苔孢子/配子放散的温度和光照强度范围分别为20~30℃和180~300μmol·m~(-2)·s~(-1)。在温度25℃、光照强度240μmol·m~(-2)·s~(-1)时,浒苔孢子/配子的放散量最大。浒苔孢子/配子萌发的适宜温度和光照强度分别为15~25℃和40~160μmol·m~(-2)·s~(-1),最适温度和光照强度分别为20℃和120μmol·m~(-2)·s~(-1)。本实验揭示了黄海绿潮优势种浒苔繁殖与生长的环境响应过程,为进一步探索黄海绿潮暴发机制提供理论依据。     

低温、低光照强度对扁浒苔生长的影响

为研究低温、低光照强度对浒苔生长和光合作用参数的影响,设置不同的光照强度和温度培养条件,对设定条件下中国漂浮浒苔种类扁浒苔(Ulvacompessa)的特定生长率进行测定,并利用Phyto-PAM荧光仪测定了低温和低光照强度条件下浒苔最大光量子产量Fv/Fm。结果表明,低温和低光照强度对扁浒苔生长的影响极显著(P0.01),且两者存在显著的交互作用(P0.05)。方差分析表明,扁浒苔在7.5℃、10℃和12.5℃条件下的特定生长率差异不显著(P0.05),但显著高于5℃的特定生长率,显著低于15℃的特定生长率(P0.05);当光照强度为10μmol(/m2·s)、20μmol(/m2·s)和30μmol(/m2·s)时,扁浒苔生长率显著高于在4μmol(/m2·s)的生长率,显著低于在40μmol(/m2·s)的生长率(P0.05)。在实验范围内随温度和光强的升高,Fv/Fm有上升趋势,但显著低于对照水平(P0.05)。综合以上结果可知,扁浒苔在温度为5℃,光照强度为4μmol(/m2·s)条件下,仍然可保持4.32%/d的特定生长率,表明扁浒苔对低温和弱光具有一定耐受性,且温度和光照共同对扁浒苔的生长产生影响,在实验范围内温度和光照强度越高,扁浒苔生长越快,最大光量子产量Fv/Fm也随之提高。     

温度对浒苔生长及不同氮源吸收特性的影响

为了评估浒苔(Ulva prolifera)对养殖废水的净化效果,本文研究了4个温度水平(22.5℃、25.5℃、28.5℃和31.5℃)和3种氮源(NH4Cl、NaNO2和NaNO3)下,浒苔对工厂化养殖水环境的适应能力及其净化效果。结果显示,96 h内4个温度处理组浒苔对总氨氮(TAN,包括NH4+-N和NH3)的平均吸收速率分别为14.65、14.88、14.48和13.53 μmol/(g?h),144 h内温度各处理组对亚硝态氮(NO2–-N)的平均吸收速率分别为11.28、10.48、9.11和8.38 μmol/(g?h),144 h内各温度处理组对硝态氮(NO3–-N)的平均吸收速率分别为9.41、8.62、8.80和7.35 μmol/(g?h);温度对浒苔的生长速率有极显著的影响(P<0.01),而氮源对浒苔的生长速率有显著影响(P<0.05);在相同氮源条件下,浒苔的生长速率随着温度的升高而逐渐降低;在相同温度条件下,氮源为氨氮(NH4+-N)时,浒苔的生长速率大于氮源为NO2–-N和NO3–-N的生长速率;温度和氮源对浒苔叶绿素a的含量影响不显著(P>0.05),氮源为NH4+-N和NO2–-N时,随着温度的升高,浒苔中叶绿素a的含量均有升高的趋势,而氮源为NO3–-N时,浒苔中叶绿素a的含量呈先降低再升高的趋势;温度和氮源对浒苔中类胡萝卜素的含量均有极显著影响(P<0.01)。随着温度的升高,各处理组浒苔中类胡萝卜素的含量均呈升高的趋势,其中,在28.5℃和31.5℃条件下,NO3–-N处理组浒苔类胡萝卜素的含量明显高于其他各处理组(P<0.05)。研究表明,温度在22.5℃~31.5℃范围内,浒苔可以有效吸收TAN、NO2–-N和NO3–-N等对虾工厂化养殖废水中的营养盐,浒苔对NH4+-N的吸收速率最大,但随着水温的升高,浒苔对NH4+-N、NO2–-N和NO3–-N的吸收速率均呈降低的趋势。     

不同氮素形式的C/N对生物絮团氨氮转化效率及营养组分的影响

碳氮比(C/N)调控是生物絮团养殖的核心技术特征,相关研究和实践中C/N中的碳和氮有不同的表征形式,本研究用溶解有机碳(dissolved organic carbon, DOC)表征碳,分别用总氮(total nitrogen, TN)、溶解无机氮(dissolved inorganic nitrogen, DIN)、总氨氮(total ammonia nitrogen, TAN)表征氮,比较了相同C/N、不同氮素形式条件下生物絮团的氨氮去除能力、基本营养组分、氮代谢相关功能基因及总异养菌数量。实验设置A组DOC/TN为20, B组DOC/DIN为20, C组DOC/TAN为20。各实验组8 h内可将10 mg/L氨氮降低到1 mg/L以下, TAN去除速率分别为(2.11±0.05)mgTAN/gTSS·h、(2.00±0.02)mgTAN/gTSS·h和(2.09±0.02)mgTAN/gTSS·h,A组显著高于B组(P<0.05),C组与A、B组无显著差异。各组絮团粗蛋白含量无显著差异,C组絮团粗脂肪含量显著高于B组和A组(P<0.05),主要氨基酸和脂肪酸组分含量...     

条浒苔海区试栽培及外界因子对藻体生长的影响

以2004年1月采自江苏如东海区紫菜栽培架上的条浒苔(Enteromorpha clathrata)为研究材料,经分离纯化培养,用条浒苔藻体放散的孢子进行采苗及其海区栽培。结果表明,条浒苔在江苏和浙江海区均能够栽培,且藻体生长速度很快。在实验室条件下,将条浒苔藻体置于不同温度组(5℃、10℃、15℃、20℃、25℃、30℃)、盐度组(0、4、8、12、16)和光照强度组(15μmol.m-2.s-1、30μmol.m-2.s-1、45μmol.m-2.s-1、60μmol.m-2.s-1),共120个温度-盐度-光强组合中培养,以研究这3个生态因子对条浒苔生长的影响。结果发现,温度、盐度和光强对条浒苔的生长影响显著(P<0.05)。条浒苔生长适宜温度为15~25℃;适宜盐度为8~12;适宜光照强度在60μmol.m-2.s-1左右。温度、盐度和光照强度三者对条浒苔生长存在显著的交互效应(P<0.05),这三者的最佳组合为温度25℃、盐度12和光强60μmol.m-2.s-1     

不同增殖方式来源的绿潮藻浒苔藻体生长及光合生理特性差异

为了探讨原位状态下不同增殖方式得到的绿潮藻浒苔生物体的差异,通过原位围隔实验,分别获取漂浮浒苔释放的孢子萌发形成的藻体(ST)和浒苔自身营养增殖得到的藻体(VT),通过比较这两种浒苔的生长、光合作用和荧光参数的不同,来估测其光合生理特性的差异。结果表明,ST的生长速率比VT显著高出61.27%,并且ST的最大光合作用速率(Pmax)、光合效率(α)和光合活性(P/R)比VT分别显著高出25.33%、14.93%和134.69%,而呼吸作用速率和光补偿点比VT显著低了45.7%和52.2%,这表明ST与VT相比具有更大的生长优势。另外,VT的相对电子传递速率(rETR)显著高于ST,在高光下尤为显著,并且VT在高光下具有更高的非光化学猝灭(NPQ)能力,这说明浒苔通过自身营养增殖产生的藻体对高光的适应能力比其由孢子萌发形成的藻体更强。     

富营养化与生长密度对绿潮藻浒苔暴发性生长机制的影响

近海海域富营养化导致的氮(N)和磷(P)增加,是绿潮藻浒苔快速增长的物质基础,而浒苔在海洋中的生长密度也会对浒苔的快速生长产生影响。通过设置不同的起始生物量浓度,研究不同生长密度与N、P对浒苔生长特性的影响,同时研究其光合生理特性的响应,以探讨浒苔暴发性增长的可能机制。结果显示,起始生物量越大,浒苔的相对生长率就越低,而N、P对浒苔的生长速率有明显的促进效应,并且N、P之间有显著的交互作用。在低密度时,+N+P处理的浒苔,最大生长速率可以达到0.75,其加倍时间仅为0.92 d,这个生长速率足可以维持浒苔在早期的暴发性增长。+N或+P都显著提高浒苔光合色素水平,这也导致+N+P组的最大光合作用速率(Pmax)比+N组、+P组和对照组分别高17%、45%和57%。富营养化和低生物密度是维持浒苔高速增长的重要条件,这也是绿潮早期暴发性增长的基础。     

浒苔中MnSOD和CAT基因克隆和表达分析

实验以大型绿藻浒苔为材料,克隆了其抗氧化系统关键酶锰超氧化物歧化酶(manganese superoxide dismutase,Mn SOD)和过氧化氢酶(catalase,CAT)基因的部分片段,利用荧光定量PCR技术研究了Mn SOD和CAT对温度和水杨酸作用的响应规律。结果获得浒苔318 bp的Mn SOD基因,该Mn SOD推测的氨基酸序列与裂片石莼Mn SOD相似性最高为89%;获得了229 bp的CAT基因,该浒苔CAT编码氨基酸序列与裂片石莼和雨生红球藻CAT序列相似性分别为99%和93%。在利用M n SOD和CAT氨基酸序列构建的系统树中,浒苔均先与裂片石莼聚类,再与其他绿藻聚在一起。不同温度对浒苔Mn SOD和CAT基因表达影响表明,35℃高温培养对浒苔Mn SOD和CAT基因表达影响最显著,其表达量分别为25℃培养的2.18倍和2.05倍;5℃低温培养次之;而15℃与25℃培养对M n SOD和CAT基因表达影响差异不显著(P0.05)。在高温35℃下,添加0.1 mmol/L水杨酸后Mn SOD和CAT基因表达量分别为各自对照的2.08倍和5.30倍,而0.01 mmol/L水杨酸添加后基因表达与对照差异不显著(P0.05)。研究表明,Mn SOD和CAT基因表达的增强不仅是浒苔对高温胁迫的响应,而且也是水杨酸缓解高温对浒苔不利影响的方式之一。     

浒苔碳汇功能评估及其扩增途径

浒苔(Ulva prolifera)不仅是绿潮暴发的主要生物种类,也是海洋中重要的碳汇生物。浒苔生长速度快、繁殖方式多样、抗逆能力强,能够在短时间内形成大规模生物量,本文在此基础上评述了中国近海浒苔生物在生长和漂移过程中的固碳特点：浒苔具有独特的高pH诱导HCO3–利用机制,可提高HCO3–的吸收效率,并促进漂浮浒苔对空气中CO2的吸收,同时,C4固碳途径增强了漂浮浒苔在高光辐射时的碳固定效率,这种多样化的碳吸收和碳固定模式,使漂浮浒苔光合固碳能力增强,漂浮状态下可以快速积累生物量。以上特点使得浒苔的固碳效率显著高于主要养殖藻类,如海带、裙带菜和紫菜等。2007年以来,黄海海域连续15年暴发世界上最大规模浒苔绿潮,最大分布面积年均在3万km2以上,最大覆盖面积年均超过500 km2,浒苔生物量年均在150万t以上,据此估算,2008―2020年净碳汇量为2.5~27.5万t,年均超过7.8万t,高于主要养殖藻类,如江蓠、紫菜和裙带菜的年均固碳量,仅次于海带的年均固碳量。浒苔巨大的生物量和强大的碳固定能力使其成为一个潜在的、不可忽视的海洋碳汇和碳储途径,建议浒苔绿潮暴发时,进一步加强浒苔打捞力度和资源化利用程度,实现碳利用和去富营养化的双赢,推动浒苔碳汇产业早日加入碳市场,使其成为一个新的低碳经济增长点。     

江苏滨海县海域漂浮浒苔光合生理特性及其固碳潜力评估

测定了江苏滨海县海域漂浮浒苔(Ulva prolifera)的光合生理特性(叶绿素荧光参数、叶绿素含量、Rubisco活性、光合速率)和组织含碳量,并分析了浒苔暴发海区水体溶解性无机碳(dissolved inorganic carbon,DIC)浓度及pH值变化特征。结果表明:1)浒苔漂到滨海海域时藻体状态良好,藻体Fv/Fm达到0.67,Yield为0.57,总光合速率为66.99μmol(C)·g-1(FW)·h-1,叶绿素a和b的含量分别为0.91 mg·g-1和0.65mg·g-1,Rubisco活性达到11.39μmol(CO2)·g-1(FW)·min-1;2)覆盖区域不同位点海水无机碳体系各参数均出现不同程度的变化,从浒苔覆盖区边缘到中心,海水中DIC和HCO3-浓度逐步降低(P0.05),而海水pH值却逐渐升高(P0.05)。3)2008~2013连续6年暴发的绿潮藻生物量累计约达9 832.8×104t湿重,可固定CO2量达1 843.67×104t,说明浒苔对DIC的吸收能力较强,能促进海水无机碳体系循环,提高海水pH值,能一定程度上增加海洋碳汇强度。     

Rare serotype occurrence and PFGE genotypic diversity of Streptococcus agalactiae isolated from tilapia in China

Previously, we reported 10 PEGE types of 85 tilapia Streptococcus agalactiae(GBS), which shifted from Streptococcus iniae in China, by using PEGE method. Presently, larger and more representative tilapia GBS were isolated, for the ?rst time in China, to characterize their serotypes and genetic diversities more precisely than had done before. 168 GBS strains were distributed in ?ve provinces of China, in which Guangdong, Guangxi and Hainan were the major ones, holding36.9%(62/168), 37.5%(63/168) and 19.6%(33/168), respectively. Serotypes, Ia, Ib and III, were observed in these strains and the most predominant one was Ia(95.2%), which mainly distributed in Guangdong, Guangxi and Hainan. Ia initially occurred in 2009, it shoot up to 32.1% in 2010,but decreased to 16.1% in 2011 before went up to 45.2% in 2012. Ib sporadically occurred during2007–2011, III onlyoccurred in 2012. 14 different PFGE types, including 4 new types(N, O,P and Q), were observed, in which B, D, F and G were the predominant types, holding 83.9%(141/168) of the total GBS strains. Ia corresponded to 11 PFGE types(A–H, N–P), in which type D predominated(51%). Ib represented 3 genotypes(I, J and Q) and III harbored only 2genotypes(N and F). Type N and Fsynchronously presented in Ia and III. In summary, the genetic diversity of tilapia GBS varied by serotypes and changed with geographical locations and years.Although Iastillpredominated, new rareserotypeIII alreadyoccurred in China.     

Growth hormone (GH) and reproduction: a review

Interaction between growth and reproduction occurs in many vertebrates and is particularly obvious at certain stages of the life cycle in fish. Endocrine interactions between the gonadotropic axis and the somatotropic axis are described, the potential role of GH being emphasised. A comparative analysis of these phenomena in mammals, amphibians and fish, suggests a specific role of GH in the physiology of puberty, gametogenesis and fertility. It also shows the original contribution made by studies on the fish model in this field of investigations.     

Purification and partial characterization of GtHs (cfLH and cfFSH) from Indian walking catfish (Clarias batrachus) (L.) and development of a homologous ELISA for cfLH

Two gonadotropins (GtH; Qa and Qb) were purified by gel filtration and ion exchange chromatography from the pituitaries of Indian walking catfish (Clarias batrachus). The presence of GtH during purification was assessed by in vitro oocyte maturation and in vivo steroidogenic activity, and their identities were determined by elution profiles, molecular weight, biological activities and yield. The molecular weights of Qa and Qb were 37 and 42 kDa, respectively, and composed of distinct subunits (Qa: 20 and 14 kDa and Qb: 26 and 18 kDa). Polyclonal antibodies raised against Qa immunostained Qa, Qb and pituitary GtH cells. A competitive Qa‐ELISA was developed whose sensitivity was 6.25 ng mL^?1 (1.25 ng well^?1) with intra‐ (3.5%) and inter‐ (12.4%) assay coefficients of variation. Displacement curves parallel to the standard were obtained with plasma and pituitary extracts of catfish, Qb and carp GtHII. The assay was validated by measuring the plasma Qa levels after LHRH treatment and in relation to ovarian growth in the female catfish during different reproductive phases. Based on the results, Qa and Qb corresponded to fish LH and FSH respectively. The findings will increase the knowledge of the mechanisms controlling fish reproduction and identification of sensitive phases in fish in captivity for hormonal manipulation.     

Controlled infection of Poecilia reticulata Peters (guppy) with Tetrahymena by immersion and intraperitoneal injection

Tetrahymena is a protozoan parasite, which infects guppy, Poecilia reticulata Peters, and causes substantial economical losses in commercial farms worldwide. Studies of guppy infected by Tetrahymena require standardized infection protocols. The LD50 for Tetrahymena infection of guppies by intraperitoneal (IP) injection was calibrated, and the level obtained was 946 parasites per fish. Guppy infection with Tetrahymena by immersion, imitating the natural route of infection via the integument, was studied under normal or stress conditions. Exposure to cold and netting (CNI) and to cold only (CI) followed by immersion exposure to 10 000 Tetrahymena per mL resulted in 22.5% and 19.2% mortality, respectively, as compared to 14.2% and 10% in groups that were netted only (NI) or non‐stressed (I). Histopathology revealed that immersion infection resulted in a systemic infection. Lysozyme levels, measured 3 weeks after infection, were significantly higher in the CNI group (288 μg per mg protein) compared with CI‐, NI‐ and I‐treated groups (94.5, 64 and 62.3 μg mg^?1, respectively). There was no evident parasite immobilization activity in body homogenates, suggesting no development of acquired immunity. Re‐infection by IP injection revealed no increase in protection in any of the treatment groups, mortality range of 56.3–75%, higher than in the non‐exposed control (40.6% mortality).     

Evaluation of a Fast Method Based on the Presence of Two Restriction Sites in the Mitochondrial ND5 (mt ND5) Gene for the Identification of Scomber Species

The purpose of this work was to evaluate the suitability of a method based on the presence of two restriction sites (for Hae III and Hindf I) in the mitochondrial NADH dehydrogenase subunit 5 (mt ND5) gene to identify Scomber species. The evaluation was performed on 144 reference and market samples by sequencing of the entire 505-bp fragment of the mt ND5 gene and of a 464-bp fragment of the Kocher fragment of the cytochrome b gene (mt Cytb). Sequence analysis of any of the two fragments allows the identification of each of the four Scomber species, but S. japonicus and S. colias had the same restriction sites at the ND5 amplicon and would not have been differentiated by this analysis. Similarly, loss of the Hae III site in some S. scombrus individuals would have misidentified them as not being Scomber. All the market products were correctly labeled except one acquired in Spain labeled as originating in the Atlantic and containing S. japonicus.     

Nitrogenous waste excretion and accumulation of urea and ammonia inChalcalburnus tarichi (Cyprinidae), endemic to the extremely alkaline Lake Van (Eastern Turkey)

The endemic, anadromous cyprinidChalcalburnus tarichi is the only fish species known to occur in alkaline Lake Van (Eastern Anatolia, Turkey). EightC. tarichi were maintained individually in Lake Van water (17 – 19°C; pH 9.8; 153 mEq·I^–1 total alkalinity; 22 total salinity) and tank water samples analyzed for 24 h in 2 to 4 h intervals. At zero time, < 1µM ammonia was present and urea was undetectable in the tank water; at 24 h, total ammonia and urea made up 114±32 and 35±25µM, respectively. Over the experimental period, ammonia-N and urea-N excretion averaged 1041±494 and 607±169moles·kg^–1 fish·h^–1, respectively. The extent of urea excretion was highly variable between specimens. Uric acid excretion was not detectable.Urea was present at high concentrations in all tissues and plasma (25 – 35moles·g^–1·ml^–1) of freshly caughtC. tarichi; total ammonia content of the tissues was by a factor of 1.9 (liver) to 3.0 (brain) lower. High arginase activity (2.4±0.2 U·min^–1·g^–1) was detected in the liver ofC. tarichi but ornithine carbamoylphosphate transferase, a key enzyme of the ornithine-urea-cycle, was absent. Ureagenesis is likely through degradation of arginine and/or uricolysis. High glutamine synthetase activity (11±0.6 U·min^–1·g^–1) and low ammonia content in brain suggest that, like other teleosts,C. tarichi has an efficient ammonia detoxification in the brain, but in no other tissue.Nitrogenous waste excretion at alkaline pH is discussed. The ability ofC. tarichi to excrete high levels of ammonia at extremely alkaline pH is unique among teleosts studied so far. The mechanism of ammonia excretion under Lake Van conditions remains to be elucidated.     

Imported ornamental fish are colonized with antibiotic‐resistant bacteria

There has been growing concern about the overuse of antibiotics in the ornamental fish industry and its possible effect on the increasing drug resistance in both commensal and pathogenic organisms in these fish. The aim of this study was to carry out an assessment of the diversity of bacteria, including pathogens, in ornamental fish species imported into North America and to assess their antibiotic resistance. Kidney samples were collected from 32 freshwater ornamental fish of various species, which arrived to an importing facility in Portland, Oregon from Colombia, Singapore and Florida. Sixty‐four unique bacterial colonies were isolated and identified by PCR using bacterial 16S primers and DNA sequencing. Multiple isolates were identified as bacteria with potential to cause disease in both fish and humans. The antibiotic resistance profile of each isolate was performed for nine different antibiotics. Among them, cefotaxime (16% resistance among isolates) was the antibiotic associated with more activity, while the least active was tetracycline (77% resistant). Knowing information about the diversity of bacteria in imported ornamental fish, as well as the resistance profiles for the bacteria will be useful in more effectively treating clinical infected fish, and also potential zoonoses in the future.     

Effect of iodophor disinfection of non‐hardened Salmo trutta eggs on their bacterial and fungus load

This study investigated the efficiency of iodophor disinfection (135 ppm active iodine for 15–30 min) of non‐hardened Salmo trutta eggs against different groups of bacteria and against fungus. Egg samples were taken from non‐disinfected and from disinfected eggs, microorganisms were cultured on specific nutrient media and their mass was measured by turbidimetric methods. Bacteria and fungus mass of non‐hardened eggs could be reduced but not eliminated by iodophor disinfection with 135 ppm active iodine for 15 min. The extent of reduction was 47–65% (Experiment 1). The efficiency of disinfection increased with disinfection time as the reduction in bacteria and fungus mass was 40–55% after 15 min and 58–74% after 30 min (Experiment 2). Disinfection efficiency of iodophor solution diluted in water (reduction 49–57%) and of iodophor solution diluted in sodium chloride solution iso‐osmolar to the oocytes (reduction 52–61%) was similar (Experiment 3). The reduction in bacteria and fungus mass was persistent as it was 39–72% lower in embryos deriving from disinfected eggs than in embryos deriving from non‐disinfected ones (Experiment 4). In conclusion, the tested disinfection method is inadequate to eliminate pathogens completely but it could positively influence immune defence of eggs and embryos.