Live vaccine trials against nocardiosis in yellowtail Seriola quinqueradiata

In an attempt to develop a vaccine against Nocardia seriolae, related species of live bacteria N. soli, N. fluminea, and N. uniformis were injected into yellowtail Seriola quinqueradiata. In addition, fish were challenged with a low virulence strain of N. seriolae to model the concept of use of a live vaccine. The fish injected with live N. soli and N. fluminea cells showed slight resistance against an artificial challenge with N. seriolae. On the other hand, the fish that survived the N. seriolae infection showed complete resistance to the N. seriolae challenge. These results suggest that protective immune responses against N. seriolae are induced in yellowtails.     

Recruitment and effects of Discocotyle sagittata (Monogenea) infection on farmed trout

Farmed rainbow trout Oncorhynchus mykiss and brown trout Salmo trutta were monitored over 3 years for infection with the blood-feeding gill fluke Discocotyle sagittata. Parasite transmission is seasonal: new infections take place during summer/autumn, and transmission is generally negligible during winter/spring. There are 2 sources of infection for naïve fish-of-the-year: limited invasion when fish are in the raceways by riverborne larvae originating external to the farm; and internally, within the farm, when 0+ fish are transferred to ponds previously occupied by older cohorts of infected fish. Thereafter, infection levels continue to increase in rainbow trout primarily through transmission within the farm. Prevalence rose to 100% in 1+ fish by the end of their second summer. In O. mykiss, mean abundance reached 194 worms/host for 1+ fish (up to 489 worms/host) and 160 worms/host for 2+ fish. By contrast, in S. trutta, parasite prevalence never exceeded 85% and, after the first year's invasions, infection levels decreased over time: in 1+ and 2+ brown trout, parasite mean abundance was < 4 (maximum 15) worms/host. We present evidence of the detrimental effects of D. sagittata on the host: high burdens are associated with pale gills, decreased body condition and host mortality. Parasite burdens become overdispersed during the warmer part of the year, as prevalence and mean abundance increase. However, the degree of parasite overdispersion decreases over winter; we cannot distinguish whether decreased aggregation is due to parasite losses from infected fish (including immune-mediated parasite mortality) or parasite-induced host mortality.     

三种鰤属鱼类肌肉质构特性及营养成分比较分析

为分析我国养殖黄条鰤(Seriola lalandi)、高体鰤(S. dumerili)、五条鰤(S. quinqueradiata)肌肉的质构特性、基本营养组成和食用价值, 采用质构分析法(TPA)和常规生化方法检测了 3 种鰤属鱼类肌肉的质构特性、粗蛋白、脂肪酸和氨基酸等成分, 并评价了营养价值。结果显示: 同等养殖条件下, 黄条鰤肌肉的硬度、胶着度、咀嚼度和回复力均显著高于高体鰤和五条鰤。黄条鰤肌肉蛋白含量(24.3%)最高, 高体鰤水分含量(70.6%)最高, 五条鰤脂肪含量(7.2%)最高, 3 种鰤属鱼类肌肉的必需氨基酸含量均优于 FAO/WHO 标准。根据 AAS 和 CS 分值, 3 种鰤属鱼类肌肉的第一限制氨基酸皆为蛋氨酸, 第二限制氨基酸皆为缬氨酸, 且肌肉鲜味氨基酸含量及其在总氨基酸中的占比均较高, 这与其味道鲜美密切相关。3 种鰤属鱼类肌肉中可检测到 20 种脂肪酸, 其中亚麻酸甲酯(C18 3n6) ∶ 只在五条鰤中检测出, 二十二碳二烯酸甲酯(C22 2) ∶ 只在高体鰤中检测出。肌肉的不饱和脂肪酸含量(65.44%~67.51%)均相对较高, 其中不饱和脂肪酸 EPA+DHA 的含量(32.50%~35.79%)优势明显。另外, 高体鰤中的常量元素含量 (5.27×10³ mg/kg)最高, 五条鰤中的微量元素含量(15.931 mg/kg)最高。本研究表明, 3 种鰤属鱼类肌肉中含有丰富的氨基酸和脂肪酸等营养物质, 是极具市场开发潜力的大洋性养殖经济鱼种, 研究结果将为我国鰤属鱼类养殖潜力评价和专用高效配合饲料的研制提供参考依据。     

Effect of temperature on growth performance of greater amberjack (SERIOLA DUMERILI Risso 1810) Juveniles

In order to successfully diversify Mediterranean aquaculture, it is necessary to determine optimum culture conditions of potential candidate species such as greater amberjack (Seriola dumerili). Among culture conditions, rearing temperature is a key factor for achieving optimum growth and maintaining fish welfare. However, little is known about the optimum culture conditions of greater amberjack (Seriola dumerili). Thus, the aim of this study was to determine the effect of three different rearing temperatures (17, 22 and 26°C) during 120 days on growth performance, body morphometry, biochemical composition, gut transit and liver morphology of greater amberjack (Seriola dumerili) juveniles. After 120 days of rearing, fish raised at 26°C showed higher (p < .05) body weight and specific growth rate than fish held at lower temperatures, as well as improved feed utilization, protein efficiency and nutrient retention percentages. Fish stomach emptying was faster (p < .05) in fish raised at 26°C than in fish held at 22°C and 17°C. Similar results were obtained for gut transit time, being gut emptying faster (p < .05) in fish reared at 26°C than in fish cultured at lower temperatures. Rearing temperature also induced changes in fish morphology which resulted in a higher (p < .05) caudal propulsion efficiency index for fish reared at 26°C. Based on these results, we conclude that greater amberjack fingerlings perform better at 26°C than at 22°C or 17°C.     

牙鲆胰岛素样生长因子结合蛋白IGFBP-1 cDNA全长的克隆及表达分析

利用RACE-PCR技术,从牙鲆(Paralichthys olivaceus)的肝脏组织总RNA中克隆得到胰岛素样生长因子结合蛋白-1(insulin-like growth factor binding protein 1, IGFBP-1)基因的全长cDNA序列,该cDNA全长为1070 bp,开放阅读框为729 bp,编码242个氨基酸。通过系统进化树分析,牙鲆IGFBP-1与鱼类IGFBP-1基因聚为一支;通过同源性比对,牙鲆IGFBP-1基因的核苷酸序列与大菱鲆同源性最高,为95%,而其推导的氨基酸序列与其它鱼类如大菱鲆、五条鰤、黄金鲈、红点鲑、鲤鱼和斑马鱼的同源性分别为89%、89%、84%、79%、67%和67%。半定量RT-PCR分析表明,牙鲆IGFBP-1基因存在母源转录本,合子基因在孵化前的胚胎阶段及早期仔鱼中仅有较低水平的表达,在后期仔鱼中表达逐渐增高;牙鲆IGFBP-1基因在肝脏中表达量最高,在胃、脾、肠、性腺、肾、鳃、脑、心脏和肌肉中也有不同程度的表达。     

Effect of different rearing conditions on body lipid composition of greater amberjack broodstock (Seriola dumerili)

The aim of this study was to assess the effect of two rearing conditions: outdoor environment with great volume tanks (500 m³) and low stocking density (~0.4 kg m^?3); and indoor environment with smaller volume tanks (10 m³) and higher stocking density (~5 kg m^?3), on muscle, liver and ovary lipid composition of Seriola dumerili broodstock born in captivity. The rearing conditions tested seem to affect the pattern of lipid body deposition in broodstock fish of S. dumerili, increasing the muscle and liver triacylglycerides (TG) accumulation of fish kept under indoor conditions, probably due to reduced energy expenditure in swimming, with some variations in the fatty acid profile that may respond to the differences in the culture conditions tested. No significant differences were found for Gonadosomatic Index (GSI) or ovary lipid deposition between groups in this study, which may suggest that the conditions tested do not have a major effect on ovary development. However, one season later the females kept under outdoor conditions released eggs spontaneously, whereas those kept under indoor conditions did not spawn, suggesting that the conditions tested actually have an effect on the broodstock's reproductive fitness. More studies are needed in order to evaluate whether the conditions tested have or not any influence on ovary development.     

Disease of rainbow trout caused by Pseudomonas luteola

Bacteria isolated from rainbow trout, Oncorhynchus mykiss, kept in a farm, in Turkey. During the outbreak, 40% of the rainbow trout (10-40 g) died. Typical clinical signs were exophthalmia, dark pigmentation, hemorrhage at the base of the pectoral, pelvic, anal fins and around the vent. Internal signs were enlarged spleen, pale liver and intestine filled with yellowish fluid. Liver, kidney and spleen of diseased fish, were aseptically streaked on Tryptic Soy Agar. After incubation, pure cultured colonies were observed and biochemically characterized with API 20 NE and other biochemical tests. Cultured bacterial 16 S rDNA gene was sequenced. Based on biochemical characteristics and sequence of 16 S rRNA, the causative bacteria were identified as Pseudomonas luteola. This study reports the first P. luteola infection in fish.     

肖四海湖五种渔具的鳜渔获结构特征及其对鳜资源的影响

为评估不同渔具对鳜资源的影响,于2007年5月和12月对长江中游浅水湖泊肖四海湖刺网、延绳钓、网簖、电拖网和电捕仪5种渔具捕获的鳜渔获物结构特征进行了调查分析.结果发现,共采集鳜样本359尾,全长分布范围为92 ～600 mm,优势全长集中于251～350 mm;体质量分布范围为10～3 380 g,优势体质量集中于300 ～500 g.种群由5个年龄组构成,2～3龄为优势龄组,占总数的74.6％.刺网、延绳钓和网簖对鳜有较强的捕捞选择性,网目大小为80 mm和100 mm刺网的鳜渔获物中2龄及以上成熟个体占总数的93.3％,个体平均体质量466 g,“标鳜”(0.4 ～0.75 kg)个体占总数量的57.0％;延绳钓捕获的鳜渔获物中2龄及以上成熟个体占总数的86.9％,“标鳜”个体占总数量的43.5％;网簖捕获的鳜渔获物中90％以上为1龄的未成熟个体.电拖网和电捕仪捕获的鳜全长范围明显较大,其渔获物以1龄和2龄个体为主.综合分析表明,刺网适于作为鳜捕捞的主要渔具,延绳钓可以作为一种鳜捕捞的辅助渔具.网簖对鳜补充群体有较大危害,不适于作为鳜的捕捞网具.电拖网和电捕仪均属于违法渔具,对鳜资源危害巨大,应该加大监管力度,严禁使用.     

黄颡鱼IgM基因的个体发生和抗体的代间传递

为了解黄颡鱼IgM基因表达的个体发生和IgM抗体代间传递机制,实验利用Real-time PCR和ELISA等技术研究了黄颡鱼IgM重链基因在卵巢、胚胎和仔鱼的表达变化,以及黄颡鱼IgM抗体在卵巢、胚胎和仔鱼中的含量变化。结果显示,黄颡鱼3~7 d仔鱼中,没有检测到IgM mRNA;14 d仔鱼IgM基因开始表达。用细菌免疫亲鱼后,对仔鱼IgM mRNA表达没有影响。黄颡鱼IgM抗体在卵巢、胚胎和仔鱼中都有分布,且呈现下降趋势,至9 d仔鱼中抗体水平最低(是卵抗体含量的0.31倍)。14 d仔鱼中IgM抗体水平上升(是9 d仔鱼抗体含量的1.6倍)。用细菌免疫亲鱼后,能显著提高胚胎、仔鱼中的IgM抗体水平,在卵中抗体含量提高了2.3倍,9 d仔鱼中提高了1.8倍。研究表明,黄颡鱼IgM抗体可以在母本和后代之间传递,早期仔鱼IgM抗体主要来自于亲本;因此,免疫亲鱼能显著增加子代的抗体水平。     

Evaluation of three seaweeds Gracilaria bursa-pastoris, Ulva rigida and Gracilaria cornea as dietary ingredients in European sea bass (Dicentrarchus labrax) juveniles

The aim of this study was to evaluate the inclusion of three seaweeds Gracilaria bursa-pastoris (GP), Ulva rigida (UR) and Gracilaria cornea (GC) as dietary ingredients on the performance, nutrient utilisation and body composition of European sea bass juveniles. Six experimental diets were formulated to replace 5% (GP-5, UR-5, and GC-5 Diets) and 10% (GP-10, UR-10 and GC-10 Diets) fish protein hydrolysate (CPSP) by each of the three seaweeds. A control diet was used, without inclusion of any seaweed. Diets were fed to duplicate groups of 25 European sea bass (Dicentrarchus labrax) juveniles (IBW = 4.7 g) for 10 weeks. Growth performance was only significantly reduced (P < 0.05) in fish fed the GC-10 diet, whereas the feed conversion ratio increased significantly in those fish. The apparent digestibility coefficients of dry matter and lipid were significantly lower in fish fed diet GC-10 relative to those fed the control diet. Carcass composition was similar among treatments, although fish fed GC-10 exhibited significantly higher ash content.The results obtained in this study suggest that the inclusion of G. bursa-pastoris (GP) and U. rigida (UR), up to 10%, can be considered as very interesting ingredients in diets for sea bass juveniles, as no negative consequences on growth performance, nutrient utilization or body composition were observed. On the other hand, the inclusion of G. cornea (GC) should be limited to 5% of the diet.     

Reproductive behaviour and early development in yellowtail kingfish (Seriola lalandi Valenciennes 1833)

The spawning behaviour of wild caught brood stock as well as early egg and larval development were studied in yellowtail kingfish (Seriola lalandi). Spawning occurred naturally in the austral spring/summer (November-February) when the seawater temperature was above 17 °C. Courtship behaviour involved one male and female, and consisted of a high-speed pursuit punctuated by stalling, nipping and touching. This lasted for approximately 0.5-1.5 h until, immediately prior to spawning, the male would nip at the female gonoduct, presumably to induce spawning. At this stage, in 50% of spawns, a second male would become involved. The release of gametes involved frenzied circling behaviour near the bottom of the tank and lasted approximately 22 s. Spawning occurred in the early daylight hours at the start of the spawning season, but shifted to around dusk in the latter part. Spawned eggs were positively buoyant, had a high fertilisation rate (> 99%), ranged 1.33-1.50 mm in diameter with a single oil droplet 0.30-0.33 mm diameter, and developed in a similar manner to that described in congenerics. Egg viability within the floating fraction was visually determined to be 74% ± 17% over the entire reproductive season. Indistinct cell margins and asymmetrical cleavage were the most common blastomere deformities observed. Egg and oil droplet volume were found to decrease by 15-20% over the spawning season, though no relationship was found between visually assessed egg viability and date. Egg incubation trials between 16 and 24 °C indicated that temperature accelerated the time to hatch by a Q₁₀ of 5.0. While larvae were found to hatch at a smaller length with a larger yolk sac and oil droplet at warmer incubation temperatures, there was little difference in the maximum larval length reached at the onset of first feeding among the rearing temperatures used. It is proposed that the reason for this was that higher incubation temperatures accelerated the hatching process faster than the rate of tissue deposition. The findings from this study are discussed in terms of the biological significance and implications for the larviculture of this species.     

Genetic structure and genetic relatedness of a hatchery stock of Senegal sole (Solea senegalensis) inferred by microsatellites

Solea senegalensis is a fish species in an early stage of domestication. A commercial hatchery in southern Spain experienced a dramatic failure in spawning and embryo survival, after incorporation into the broodstock of fish reared in their hatchery. To assess the impact of management on the stock quality, a population genetics study was made of adult broodstock present at this hatchery in 2000. The broodstock was composed of a group of fish of wild origin and two more groups from F₁ progeny. A set of eight microsatellite loci was used to compare genetic variability among the three groups and to establish the relationships between pairs of individuals within each group.Individuals from F₁ origin showed a substantial reduction in genetic variability when compared to those from the wild. There was a greater than 50% decrease in the number of alleles per locus corrected by sample size and 16% and 26% reductions in H_e values in the two F₁ groups, respectively. The degree of genetic relationship between individuals, evaluated by the relatedness estimator, rxy, and by graphical approaches, revealed a high proportion, almost 75%, of full-sibs and half-sibs in both F₁ groups, which might account for the reduction in overall variability. The high proportion of siblings also suggests a reproductive pattern in this stock in which few individuals breed successfully. This mating behaviour, along with factors inherent to selection and management systems, might be responsible for the negative effects on the reproductive capacity of the stock.     

Shading of net cage is an effective control measure against skin fluke Neobenedenia girellae infection in chub mackerel Scomber japonicus

The skin fluke Neobenedenia girellae has become a serious problem in Japan since the 1990s. Present control methods focus on the removal of the attached parasite and these post-infection treatments are often labor intensive, time consuming, and/or stressful to fish. Chub mackerel Scomber japonicus are highly susceptible to N. girellae. However, because of their sensitive nature, bath treatments may cause mortality. In this study, we investigated the efficacy of cage shading to reduce skin fluke infection and the frequency of conventional post-infection treatments. Juvenile mackerel were reared in cages with or without shade for 3 months and their skin fluke infections were monitored. We performed either freshwater baths or oral administration of praziquantel if fluke intensity exceeded the given criteria. In unshaded cages, 3 total bath treatments or 6 total drug treatments were conducted. In contrast, no treatment was required for the shaded cage. The overall fluke intensity in the shaded cage was less than half that of the unshaded cages, despite the lack of treatments. This study demonstrated for the first time the practical use of shading in fish farms to reduce skin fluke infection.     

黄条鰤线粒体全基因组测序及结构特征分析

通过二代测序、软件拼接获得中国黄海海域黄条鰤(Seriola aureovittata)线粒体基因组全序列。其序列全长为16609 bp,碱基组成分别为A(26.68%)、G(17.84%)、C(30.12%)和T(25.36%);共有13条蛋白编码基因, 22个tRNA基因, 2个rRNA基因,除NAD6、trnQ、trnA、trnN、trnC、trnY、trnS、trnE、trnP外,其余基因均在H链上编码。黄条鰤线粒体基因组全序列与蛋白编码基因的A+T含量分别为52.05%和51.085%,具有明显的AT偏好性。线粒体基因中存在2个散在重复序列,分别位于NAD1基因序列正义链的中游和COX2基因序列反义链的上游。在其22个tRNA基因中,除了tRNAGly外,均具有典型的三叶草二级结构。黄条鰤线粒体基因组的蛋白编码基因起止位点与密码子除COX1、NAD5外,均与日本海域出产的黄条鰤(Seriola lalandi)完全吻合,且COX1、NAD5基因皆短于日本黄条鰤;两者间存在一定的遗传差异。基于18种隶属于13属的鲹科鱼类线粒体基因组全序列构建系统发生树,可知小甘鲹(Seriolina nigrofasciata)、黄条鰤、五条鰤(Seriola quinqueradiata)、高体鰤(Seriola dumerili)、长鳍鰤(Seriola rivoliana)同属一近支,且黄条鰤与五条鰤亲缘关系最近,与小甘鲹进化距离较远。     

Effect of Continuous Food Supply on the Oxygen Consumption of Young Mediterranean Yellowtail (Seriola dumerili Risso, 1810)

The aim of this work was to investigate the effect of a continuous food supply on the oxygen consumption of young Mediterranean yellowtails (Seriola dumerili Risso, 1810) during the photophase period (14LL:10DD). Four groups with six fish each, weighing individually approx. 600 g, were distributed into open 2 m³ quadrangular fibreglass tanks, and fed with commercial pellets for 60 days. Two of the tanks received a continuous feed supply by means of automatic band feeders (ABF), whilst fish in the other two tanks were fed by hand, twice a day. To improve accuracy in calculating oxygen consumption, the effect of oxygen diffusion between the air and the water was determined by the use of a control tank of similar characteristics, containing no fish. Oxygen consumption instantaneous rate was calculated by the difference between dissolved oxygen concentrations in tanks with and without fish, multiplied by the water flow and divided by the estimated total fish biomass in each tank. In both feeding regimes, an increase in the oxygen consumption levels after feeding commencement was observed. The amplitude, however, was lower and the duration of feeding effect was higher in the group fed by ABF.     

Pseudomonas putida infection in rainbow trout

A disease outbreak occurred during the Spring of 2006 among rainbow trout, Oncorhynchus mykiss, kept in growout tanks. During the outbreak 35% of the rainbow trout died. Typical clinical signs were ulceration on the dorsal surface of the fish. Based on biochemical characteristics and sequence of 16S rRNA, the causative bacteria isolated from diseased fish were identified as Pseudomonas putida. Histologically, skin ulcer initially was characterized by sloughed off epithelia and epithelial necrosis. In the advanced form of the disease, almost all layers of the skin down to the epidermis were lost. Outside of Japan, this study reports the first P. putida infection in fish.     

Mugil curema in Argentinean waters: Combined morphological and molecular approach

A previously unidentified mullet species (Mugil sp.) from Mar Chiquita lagoon, Argentina, was compared with two candidate species Mugil curema and M. cephalus using 12 truss landmarks and three mitochondrial DNA genes (12S rRNA, cytochrome b and COI). Both procedures confirmed that the Argentinean fish were white mullet M. curema. Morphological comparisons clearly segregated M. curema from M. cephalus samples in PCA plot. Similarly, Argentinean haplotypes at each gene overlapped with M. curema reference samples while M. cephalus haplotypes remained distinct. These combined data provide a valuable baseline for further investigations on the geographic distribution of this commercially important species group.     

Identification and phylogenetic analysis of Vibrio vulnificus isolated from diseased Trachinotus ovatus in cage mariculture

In October 2005, an infectious disease occurred as outbreaks of high mortality within one week, which was responsible for important economic losses in intensive culture of Trachinotus ovatus around the Gulf Coast of Yangjiang city (Guangdong Province, China). The present study documented the causative agent of the diseased fish suffering from external haemorrhages and ulcers, haemorrhagic gills, livers and intestine. The strain was isolated from the diseased fish by ZoBell 2216 E agar and confirmed the pathogenicity by challenge experiments. Biochemically, the strain showed properties and biochemical characteristics similar to Vibrio vulnificus, with the exception of several atypical biochemical characteristics, especially the sucrose-positive. Meanwhile, sequencing of the 16S rRNA gene and the hemolysin gene revealed that the isolated strain was highly homogeneous with V. vulnificus. To sum up, the isolated strain was confirmed as V. vulnificus on the basis of phenotypic characteristics, phylogenetic analysis of the 16S rRNA sequences and sequence analysis of the hemolysin gene. To our knowledge, this is the first report on the V. vulnificus infection in T. ovatus.     

拟态弧菌的绿色荧光蛋白标记及其在感染草鱼体内的动态分布

为了示踪研究拟态弧菌感染草鱼的动态过程,将增强型绿色荧光蛋白编码基因EGFP克隆至质粒pBAD24,并转化到拟态弧菌04-14菌株构建荧光标记重组菌.重组菌经阿拉伯糖诱导后,能高效表达EGFP蛋白;荧光显微镜观察和流式细胞仪检测均发现重组菌能够发出明显的绿色荧光信号,且传至30代后质粒稳定率仍为100％;生物学特性检测结果显示,与野生株相比,重组菌的形态、生长特性和细胞黏附性均未发生明显改变.用标记重组菌浸泡感染草鱼,定点采集鳃、肠道、肌肉、头肾、脾脏和肝脏,借助荧光信号检测4d内细菌在不同组织脏器中的动态分布.结果发现感染4h后即可在肠道和鳃中检测到绿色荧光信号,标记菌检出量分别为3.60×108和2.36×106 CFU/g,直至10 h,其含量无明显变化,12 h后含菌量逐渐下降,但持续存在直至鱼死亡.标记菌在肌肉、头肾、脾脏和肝脏中呈现相似的动力学,感染24 h后才检测到荧光信号,24～ 85 h时间段含菌量呈现先增加后下降的变化,48 h达到峰值,检出量分别为9.58×104(肌肉)、8.75×104(头肾)、1.50×104(脾脏)和4.50×104 CFU/g(肝脏),但均低于肠道中的检出量,结果表明肠道是拟态弧菌黏附定植与繁殖的主要靶器官.     

水族箱气单胞菌的鉴定及致病特性分析

为了确定南京市某渔场发病鱼感染的病原,本研究采集患鱼脏器,采用平板培养、生化实验和特异性gyrB基因扩增测序等方法进行细菌的分离鉴定;利用PCR技术检测分离株毒力基因的分布,分析其生物学特性,并进一步通过动物实验确定菌株致病力.结果分离到1株水族箱气单胞菌,命名为LK-25.该菌株携带5种主要毒力基因:气溶素(aer)、细胞毒性肠毒素(act)、细胞兴奋性肠毒素(alt)、温敏胞外蛋白酶(epr)和丝氨酸蛋白酶(ahp),其溶血性和溶蛋白能力较强,对斑马鱼的半数致死量为1.02×103 CFU/尾,确定为强毒株.进化树分析表明,水族箱气单胞菌与嗜水气单胞菌达卡亚种亲缘关系较近.本研究在国内首次报道发现水族箱气单胞菌,为进一步预防该菌所引起的相关疾病的发生和传播提供理论基础.