Biomass and carbon storage of Gracilariopsis lemaneiformis （Rhodophyta） in Zhanshan Bay,Qingdao, China

Marine macroalgae can absorb carbon and play an important role in carbon sequestration. As an important economic macroalga, Gracilariopsis lemaneiformis has the potential to significantly affect carbon absorption and storage in wave-sheltered intertidal reef systems. However, detailed knowledge on seasonal biomass changes and carbon storage of G. lemaneiformis is lacking, especially in many small and scattered ecosystems. Considering the influence of human activities on wild distribution of G. lemaneiformis, the understanding of seasonal dynamics of an economically important species in nature is necessary. In this study, we first investigated seasonal variations in biomass, coverage area, and carbon storage during low tide from August 2011 to July 2012 in Zhanshan Bay, Qingdao, China. Furthermore, we estimated the carbon storage potential of wild G. lemaneiformis using light use efficiency(LUE). The results show that the standing biomass and coverage area changed significantly with season. However, seasonal variations in carbon content and water content were not obvious, with an average content of 35.1% and 83.64%, respectively. Moreover, carbon storage in individual months varied between 0.67 and 47.03 g C/m 2, and the value of carbon storage was the highest in August and June and the lowest in February. In Zhanshan Bay, LUE of G. lemaneiformis was only 0.23%. If it is increased to the theoretical maximum(5%–6%), the carbon storage will have an increase of at least 21 times compared with the current, which suggested that carbon storage of wild G. lemaneiformis had a high enhancement potential. The study will help to assess a potential role of G. lemaneiformis in reducing atmospheric CO2.     

Analysis of DNA Methylation of <Emphasis Type="Italic">Gracilariopsis lemaneiformis</Emphasis> Under Temperature Stress Using the Methylation Sensitive Amplification Polymorphism (MSAP) Technique

Gracilariopsis lemaneiformis is an economically important agarophyte, which contains high quality gel and shows a high growth rate. Wild population of G. lemaneiformis displayed resident divergence, though with a low genetic diversity as was revealed by amplified fragment length polymorphism (AFLP) and simple sequence repeat (SSR) analyses. In addition, different strains of G. lemaneiformis are diverse in morphology. The highly inconsistence between genetic background and physiological characteristics recommends strongly to the regulation at epigenetic level. In this study, the DNA methylation change in G. lemaneiformis among different generation branches and under different temperature stresses was assessed using methylation sensitive amplified polymorphism (MSAP) technique. It was shown that DNA methylation level among different generation branches was diverse. The full and total methylated DNA level was the lowest in the second generation branch and the highest in the third generation. The total methylation level was 61.11%, 60.88% and 64.12% at 15°C, 22°C and 26°C, respectively. Compared with the control group (22°C), the fully methylated and totally methylated ratios were increased in both experiment groups (15°C and 26°C). All of the cytosine methylation/demethylation transform (CMDT) was further analyzed. High temperature treatment could induce more CMDT than low temperature treatment did.     

Comparison in copper accumulation and physiological responses of Gracilaria lemaneiformis and G. lichenoides (Rhodophyceae)

Heavy metal pollution has become a worldwide problem in aquaculture. We studied copper (Cu2+ ) accumulation and physiological responses of two red algae Gracilaria lemaneiformis and Gracilaria lichenoides from China under Cu2+ exposure of 0-500 μg/L in concentration. Compared with G. lemaneiformis, G. lichenoides was more capable in accumulating Cu2+ , specifically, more Cu2+ on extracellular side (cell wall) than on intracellular side (cytoplasm) and in cell organelles (especially chloroplast, cell nucleus, mitochondria, and ribosome). In addition, G. lichenoides contained more insoluble polysaccharide in cell wall, which might promote the extracellular Cu2+ -binding as an efficient barrier against metal toxicity. Conversely, G. lemaneiformis was more vulnerable than G. lichenoides to Cu 2+ toxin for decreases in growth, pigment (chlorophyll a, chlorophyll b, phycobiliprotein, and β-carotene) content, and photosynthetic activity. Moreover, more serious oxidative damages in G. lemaneiformis than in G. lichenoides, in accumulation of reactive oxidative species and malondialdehyde, and in electrolyte leakage, because of lower antioxidant enzyme (superoxide dismutase and glutathione reductase) activities. Therefore, G. lichenoides was less susceptible to Cu2+ stress than G. lemaneiformis.     

Optimization of conditions for tetraspore release and assessment of photosynthetic activities for different generation branches of Gracilaria lemaneiformis Bory

Gracilaria lemaneiformis Bory is an economically important alga that is primarily used for agar production. Although tetraspores are ideal seeds for the cultivation of G. lemaneiformis, the most popular culture method is currently based on vegetative fragments, which is labor-intensive and time-consuming. In this study, we optimized the conditions for tetraspore release and evaluated the photosynthetic activities of different colonies formed from the branches of G. lemaneiformis using a PAM (pulse-amplitude-modulated) measuring system. The results showed that variations in temperature and salinityhad significant effects on tetraspore yield. However, variations in the photon flux density (from 15 μmol m⁻² s⁻¹ to 480 μmol m⁻² s⁻¹) had no apparent effect on tetraspore yield. Moreover, the PAM-parameters Y(I), Y(II), ETR(I), ETR(II) and F _v/F _m of colonies formed from different branches showed the same trend: parameter values of first generation branches>second generation branches>third generation branches. These results suggest that the photosynthetic activities of different colonies of branches changed with the same trend. Furthermore, photosynthesis in G. lemaneiformis was found to be involved in vegetative reproduction and tetraspore formation. Finally, the first generation branches grew slowly, but accumulated organic compounds to form large numbers of tetraspores. Taken together, these results showed that the first generation branches are ideal materials for the release of tetraspores.     

Inhibition of photosynthesis in the microalga Chaetoceros curvisetus （Bacillariophyta） by macroalga Gracilaria lemaneiformis （Rhodophyta）

We investigated the effects of dried macroalga Gracilaria lemaneiform （Rhodophyta） on photosynthesis of the bloom-forming microalga Chaetoceros curvisetus. C. curvisetus was cultured with different amounts of dried G. lemaneiformis under controlled laboratory conditions. We measured the photosynthetic oxygen evolution rate and established the chlorophyll a fluorescence transient （OJIP） curve coupled with its specific parameters. We observed concentration-dependent and time-dependent relationships between dried G. lemaneiformis and inhibition of photosynthesis in C. curvisetus. Co-culture with dried G. lemaneiformis also resulted in a decrease in the light-saturated maximum photosynthetic oxygen evolution rate （P~ax） in C. curvisetus, and a decrease in the OJIP curve along with its specific parameters; the maximum photochemical efficiency of PSII （FJFm）, the amount of active PSII reaction centers per excited cross section at t=0 and t=--tFM （RC/CS0 and RC/CSm, respectively）, the absorption flux per excited cross section at t=0 （ABS/ CS0）, and the efficiency with which a trapped exciton moves an electron into the electron transport chain （~u0）. The dark respiration rate （Rd） increased in C. curvisetus co-cultured with dried G. lemaneiformis. The JIP-test and the oxygen evolution results indicated that dried G. lemaneiJbrmis decreased the number of active reaction centers, blocked the electron transport chain, and damaged the oxygen-evolving complex of C. curvisetus. This result indicated that dried fragments of G. lemaneiformis could effectively inhibit photosynthesis of C. curvisetus, and thus, could serve as a functional product to control and mitigate C. curvisetus blooms.     

Morphological study of the genus <Emphasis Type="Italic">Herposiphonia</Emphasis> (Rhodophyta,Rhodomelaceae) on the coast of eastern Guangdong,China, with a description of <Emphasis Type="Italic">H. pinnata</Emphasis> sp. nov.

We present a taxonomic study of taxa of the red algae genus Herposiphonia (Rhodophyta, Rhodomelaceae), collected from the coast of eastern Guangdong, China. We made detailed morphological studies and considered recent taxonomic criteria for species delimitation, and are making the first report of five different species on the coast of Guangdong, including a new species. The species identified were H. caespitosa Tseng, H. hollenbergii Dawson, H. pecten - veneris (Harvey) Falkenberg, H. subdisticha Okamura and H. pinnata Ding and Tan sp. nov. H. pinnata sp. nov. is characterized by bright green thalli; most parts of the feathery thalli are free of the substratum; determinate branches and indeterminate branches are arranged in a chaotic sequence; the primary axis has bare segments; the determinate branch has 9–11 periaxial cells per segment; vegetative trichoblasts are abundant; and tetrasporangia are formed on the middle of the determinate branch with 1–8 successive segments in a single rectilinear series. This paper is also the first record of sporophyte plants of H. pecten-veneris.     

Development of a SCAR marker for male gametophyte of Gracilariopsis lemaneiformis based on AFLP technique

The red alga Gracilariopsis lemaneiformis （Bory） is an economically valuable macroalgae. As a means to identify the sex of immature Gracilariopsis lemaneiformis, the amplified fragment length polymorphism （AFLP） technique was used to search for possible sex- or phase-related markers in male gametophytes, female gametophytes, and tetrasporophytes, respectively. Seven AFLP selective amplification primers were used in this study. The primer combination E-TG/M-CCA detected a specific band linked to male gametophytes. The DNA fragment was recovered and a 402-bp fragment was sequenced. However, no DNA sequence match was found in public databases. Sequence characterized amplified region （SCAR） primers were designed from the sequence to test the repeatability of the relationship to the sex, using 69 male gametophytes, 139 female gametophytes, and 47 tetrasporophytes. The test results demonstrate a good linkage and repeatability of the SCAR marker to sex. The SCAR primers developed in this study could reduce the time required for sex identification of Gracilariopsis lemaneiformis by four to six months. This can reduce both the time investment and number of specimens required in breeding experiments.     

Ecological studies ofGracilaria asiatica andGracilaria lemaneiformis in Zhanshan Bay,Qingdao

Natural populations ofG. asiatica Zhang & Xia andG. lemaneiformis (Bory) Weber van Bosse were studied during 1984 and 1986 in Zhanshan Bay, Qingdao (36°4'N, 120° 21'E). Rapid growth (length, weight) of these plants occurred between mid-May and late June (water temperatures, 15–20°C). The major epiphyte ofG. asiatica wasEnteromorpha linza, whilePunctaria latifolia was the major epiphyte ofG. lemaneiformis. Epiphytism declined throughout early summer, and epiphytes were rare after mid-July (1984); they did not reappear in late summer, although macrophyte growth declined abruptly after early July. Populations ofG. asiatica varied during late spring-early summer between adjacent sandy and rocky portions of the intertidal zone; plants at the sandy site were larger and epiphyte-free. Amphipod densities were low on both species ofGracilaria, but the most abundant species wereAmpithoe lacertosa, Caprella equilibra, C. krøyeri, C. scauraandPontogeneia rostrata. Additional information on general community structure is provided for theG. asiatica zone.     

An N-shape thermal front in the western South Yellow Sea in winter

An N-shape thermal front in the western South Yellow Sea (YS) in winter was detected using Advanced Very High Resolution Radiation (AVHRR) Sea Surface Temperature data and in-situ observations with a merged front-detecting method. The front, which exists from late October through early March, consists of western and eastern wings extending roughly along the northeast-southwest isobaths with a southeastward middle segment across the 20–50 m isobaths. There are north and south inflexions connecting the middle segment with the western and eastern wings, respectively. The middle segment gradually moves southwestward from November through February with its length increasing from 62 km to 107 km and the southern inflexion moving from 36.2°N to 35.3°N. A cold tongue is found to coexist with the N-shape front, and is carried by the coastal jet penetrating southward from the tip of the Shandong Peninsula into the western South YS as revealed by a numerical simulation. After departing from the coast, the jet flows as an anti-cyclonic recirculation below 10 m depth, trapping warmer water originally carried by the compensating Yellow Sea Warm Current (YSWC). A northwestward flowing branch of the YSWC is also found on the lowest level south of the front. The N-shape front initially forms between the cold tongue and warm water involved in the subsurface anti-cyclonical recirculation and extends upwards to the surface through vertical advection and mixing. Correlation analyses reveal that northerly and easterly winds tend to be favorable to the formation and extension of the N-shape front probably through strengthening of the coastal jet and shifting the YSWC pathway eastward, respectively.     

New record of <Emphasis Type="Italic">Lobophora rosacea</Emphasis> (Dictyotales; Phaeophyceae) from the South China Sea

Lobophora rosacea C.W. Vieira, Payri et De Clerck is reported from the South China Sea for the first time. Our specimens are very similar to L. rosacea recently described from New Caledonia, not only in morphology but also in rbcL and cox3 gene sequences. The fan-shaped thallus grows erectly, attaching to the substrate by a basal holdfast. The thallus is composed of a single layer of large medullary cells and three to four layers of cortical cells on both sides of the medulla. Mature sporophytes are detected, with sporangium sori scattered on both surfaces of the thallus.     

Genome size of <Emphasis Type="Italic">Alexandrium catenella</Emphasis> and <Emphasis Type="Italic">Gracilariopsis lemaneiformis</Emphasis> estimated by flow cytometry

Flow cytometry(FCM) technique has been widely applied to estimating the genome size of various higher plants. However, there is few report about its application in algae. In this study, an optimized procedure of FCM was exploited to estimate the genome size of two eukaryotic algae. For analyzing Alexandrium catenella, an important red tide species, the whole cell instead of isolated nucleus was studied, and chicken erythrocytes were used as an internal reference. The genome size of A. catenella was estimated to be 56.48 ± 4.14 Gb(1C), approximately nineteen times larger than that of human genome. For analyzing Gracilariopsis lemaneiformis, an important economical red alga, the purified nucleus was employed, and Arabidopsis thaliana and Chondrus crispus were used as internal references, respectively. The genome size of Gp. lemaneiformis was 97.35 ± 2.58 Mb(1C) and 112.73 ± 14.00 Mb(1C), respectively, depending on the different internal references. The results of this research will promote the related studies on the genomics and evolution of these two species.     

Preference of the herbivorous marine teleost Siganus canaliculatus for different macroalgae

The decomposition of a large amount of unexploited macroalgal resource along the coast of China often results in heavy environmental pollution. In order to pave a way of using macroalgae as the dietary ingredient of rabbitfish Siganus canaliculatus, one of a few farmed herbivorous marine teleosts in China, its preference （feeding selectivity） for different macroalgae was determined in this study. Seven seaweed species abundantly inhabiting the coast of east Guangdong Province were exposed simultaneously to rab-bitfish juveniles in laboratory （multiple-choice feeding） with their content and absolute intake assayed. It was found that the most preferred algae were Ulva prolifera, Gracilaria lemaneiformis and Chaetomorpha linum, less preferred algae were U. pertusa and Porphyra haitanensis, and least preferred ones were Sargassum fusiforme and Corallina sessilis. Such an order did not change when one to four relatively preferred seaweeds were removed. The preferred seaweeds were richer in protein and soluble sugar thus higher in energy than the least preferred. In addition, this fish was found to favor filamentous and flat algae rather than calcified ones. Ac-cordingly, the richness of nutrients and morphological characteristics determined the preference of S. canaliculatus for tested macro-algae.     

Genetic transformation of <Emphasis Type="Italic">Nannochloropsis oculata</Emphasis> with a bacterial phleomycin resistance gene as dominant selective marker

The gene ble from Streptoalloteichus hindustanus is widely used as a selective antibiotic marker. It can control the phleomycin resistance, and significantly increase the tolerance of hosts to zeocin. The unicellular marine microalga Nannochloropsis oculata is extremely sensitive to zeocin. We selected ble as the selective marker for the genetic transformation of N. oculata. After the algal cells at a density of 2×10⁷ cells mL^?1 was digested with 4% hemicellulase and 2% driselase for 1 h, the protoplasts accounted for 90% of the total. The ble was placed at the downstream of promoter HSP70A-RUBS2 isolated from Chlamydomonas reinhardtii, yielding a recombinant expression construct pMS188. The construct was transferred into the protoplasts through electroporation (1 kV, 15 μS). The transformed protoplasts were cultured in fresh f/2 liquid medium, and selected on solid f/2 medium supplemented with 500 ng mL^?1 zeocin. The PCR result proved that ble existed in the transformants. Three transformants had been cultured for at least 5 generations without losing ble. Southern blotting analysis showed that the ble has been integrated into the genome of N. oculata. The ble will serve as a new dominant selective marker in genetic engineering N. oculata.     

<Emphasis Type="Italic">Pseudolella major</Emphasis> sp. nov. (Axonolaimidae,Nematoda) from the intertidal zone of the East China Sea

A new species of free-living nematode, Pseudolella major sp. nov. is described from the intertidal mudflats of Ximen Island in the East China Sea. Pseudolella major sp. nov. is characterized as follows: body length ~1 300 μm; short cephalic setae, 2–3 μm long; amphidial fovea loop-shaped with elongated ventral branch; ventral branch 50–60 μm long and extending past the base of buccal cavity; buccal cavity cylindrical with arching or spherical base; three heavy outwardly-curving odontia with blunt tips located at anterior of buccal cavity; pharynx short with posterior bulb; ventral gland cell elongate oval-shaped, located posterior to pharyngo-intestinal junction; excretory pore situated at the level of the middle of buccal cavity; tail conical to cylindrical; spicules arched, with double-cephalate proximal end; gubernaculum with a strong dorsal caudal apophysis.     

Magnitude and forming factors of mass elevation effect on Qinghai-Tibet Plateau

Mass elevation effect(MEE) refers to the thermal effect of huge mountains or plateaus, which causes the tendency for temperature-related montane landscape limits to occur at higher elevations in the inner massifs than on their outer margins. MEE has been widely identified in all large mountains, but how it could be measured and what its main forming-factors are still remain open. This paper, supposing that the local mountain base elevation(MBE) is the main factor of MEE, takes the Qinghai-Tibet Plateau(QTP) as the study area, defines MEE as the temperature difference(ΔT) between the inner and outer parts of mountain massifs, identifies the main forming factors, and analyzes their contributions to MEE. A total of 73 mountain bases were identified, ranging from 708 m to 5081 m and increasing from the edges to the central parts of the plateau. Climate data(1981–2010) from 134 meteorological stations were used to acquire ΔT by comparing near-surface air temperature on the main plateau with the free-air temperature at the same altitude and similar latitude outside of the plateau. The ΔT for the warmest month is averagely 6.15℃, over 12℃ at Lhatse and Baxoi. A multivariate linear regression model was developed to simulate MEE based on three variables(latitude, annual mean precipitation and MBE), which are all significantly correlated to ΔT. The model could explain 67.3% of MEE variation, and the contribution rates of three independent variables to MEE are 35.29%, 22.69% and 42.02%, respectively. This confirms that MBE is the main factor of MEE. The intensive MEE of the QTP pushes the 10℃ isotherm of the warmest month mean temperature 1300–2000 m higher in the main plateau than in the outer regions, leading the occurrence of the highest timberline(4900 m) and the highest snowline(6200 m) of the Northern Hemisphere in the southeast and southwest of the plateau, respectively.     

Three tropical seagrasses as potential bio-indicators to trace metals in Xincun Bay, Hainan Island, South China

Concentrations of the trace metals Cu,Cd,Pb,and Zn were measured in seawater,rhizosphere sediments,interstitial water,and the tissues of three tropical species of seagrasses (Thalassia hemprichii,Enhalus acoroides and Cymodocea rotundata) from Xincun Bay of Hainan Island,South China.We analyzed different environmental compartments and the highest concentrations of Pb and Zn were found in the interstitial and seawater.The concentrations of Cd and Zn were significantly higher in blades compared with roots or rhizomes in T.hemprichii and E.acoroides,respectively.A metal pollution index (MPI) demonstrated that sediment,interstitial water,and seagrasses in the sites located nearest anthropogenic sources of pollution had the most abundant metal concentrations.There was obvious seasonal variation of these metals in the three seagrasses with higher concentrations of Cu,Pb and Zn in January and Cd in July.Furthermore,the relationships between metal concentrations in seagrasses and environmental compartments were positively correlated significantly.The bioconcentration factors (BCF) demonstrated that Cd from the tissues of the three seagrasses might be absorbed from the sediment by the roots.However,for C.rotundata,Zn is likely to be derived from the seawater through its blades.Therefore,the blades of T.hemprichii,E.acoroides and C.rotundata are potential bio-indicators to Cd content in sediment,and additionally Zn content (C.rotundata only) in seawater.     

Production of dimethylsulfide and acrylic acid from dimethylsulfoniopropionate during growth of three marine microalgae

We measured the concentrations of dimethylsulfide （DMS）, acrylic acid （AA）, and dimethylsulfoniopropionate （DMSP） during growth of three microalgae： Prorocentrum micans, Gephyrocapsa oceanica, and Platymonas subcordiformis. The DMSP, AA, and DMS concentrations in culture media varied significantly among algal growth stages, with the highest concentrations in the late stationary growth stage or the senescent stage. In the stationary growth stage, the average DMSP concentration per cell in P. micans （0.066 5 pmol/cell） was 1.3 times that in G. oceanica （0.049 5 pmol/ cell） and 20.2 times that in P. subcordiformis （0.003 29 pmol/cell）. The average concentrations of AA were 0.044 6, 0.026 9, and 0.003 05 pmol/cell in P. micans, G. oceanica, and P. subcordiformis, respectively, higher than the concentrations of DMS （0.272, 0.497, and 0.086 2 fmol/cell, respectively）. There were significant positive correlations between cell density and AA, DMSP, and DMS concentrations. The ratios of DMS/AA and AA/（DMSP＋AA） in the three algae differed significantly over the growth cycle. In all three microalgae, the DMS/AA ratios were less than 25% during the growth period, suggesting that the enzymatic cleavage pathway, which generates DMS, was not the main DMSP degradation pathway. The changes in the DMS/AA ratio indicated that there was a higher rate of enzymatic breakdown of DMSP in the early growth period and a lower rate during senescence. In all three microalgae, the AA/（DMSP＋AA） ratio （degradation ratio of DMSP） decreased during the exponential growth phase, and then increased. The variations in these ratios can approximately indicate the cleavage mechanism of DMSP at different stages of algal growth.     

Phylogenetic analysis of epibacterial communities on the surfaces of four red macroalgae

Macroalgal surfaces are prone to being attached by bacteria. Epibacterial community structures on marine macroalgae are host-specific but temporally and spatially variable. In this study, we investigated the structure of epibacterial communities on the surfaces of four red macroalgae, Gracilaria lemaneiformis, Gloiopeltis furcata, Mazzaella sp. and Porphyra yezoensis, by analyzing the sequences of 16 S r RNA gene libraries. Healthy individuals of all macroalgae species were collected in winter from a farm at Dalian, China. The results showed that the epibacterial communities were mainly dominated by α-Proteobacteria, γ-Proteobacteria and Bacteroidetes. Deinococcus-Thermus, Spirochaetes and ε-Proteobacteria were also found. The majority of cloned sequences shared the greatest similarity to those of culturable organisms. A large portion of sequences from the α-Proteobacteria homed in Roseobacter clade, i.e., genera Ahrensia, Roseovarius, Litoreibacter, Octadecabacter, Thaiassobacter and Sulfitobacter, while members of Bacteroidetes mainly belonged to family Flavobacteriaceae. The cloned sequences could be separated into 66 OTUs at 0.01 distance value, and rare common OTUs were found among libraries. At genus level, Pseudoalteromonas dominated Gr. lemaneiformis and Gl. furcata libraries, accounting for 72.2% and 47.3%, respectively. Sulfitobacter dominated P. yezoensis library, accounting for 35.4%. A previously undefined cluster within Deinococcus-Thermus dominated Mazzaella sp. library, accounting for 24.6% of the all. These results indicated that a broad range of bacteria inhabited the surfaces of these macroalgae.     

Floc meal as potential substitute for soybean meal in tilapia diets under biofloc system conditions

This study was conducted to evaluate the effect of replacing soybean meal (SBM) by low protein floc meal (LPFM; 24% CP) in tilapia diets on growth performance, feed utilization and fish chemical composition. Three isonitrogenous and isocaloric diets were formulated; control diet (C; without LPFM), FM ₂₅ (25% of SBM protein was substituted by LPFM) and FM ₅₀ (50% of SBM protein was substituted by LPFM). Nine 55l circular plastic tanks were stocked by 12 fish to form three experimental groups. No differences in tilapia performance were observed between the control and the FM ₂₅ diet. Chemical composition of fish did not differ significantly among treatments except for protein and selenium contents where the highest values were recorded in the control treatment. The highest mineral content was recorded in floc collected from the control tanks, while calcium content showed its highest value in floc collected from FM ₅₀ effluent. These data indicate that replacing soybean meal with LPFM up to 25%, had no negative effect on growth performance and potentially may improve the system sustainability. Meanwhile, the adverse effect of more inclusion of LPFM in tilapia diet may be attributed to the higher content of ash. In other word, minerals seem to cause more energy utilization in fish fed floc meal since it is needed to maintain osmotic homeostasis.

     

Effect of dietary olaquindox on the growth of large yellow croaker (Pseudosciaena crocea R.) and the distribution of its residues in fish tissues

Olaquindox(OLA), one of quinoxaline-N, N-dioxides, has been put under ban. However it was used as a medicinal feed additive early; it promotes the growth of livestock and prevents them from dysentery and bacterial enteritis. In this study, we evaluated the effect of dietary OLA on the growth of large yellow croaker(Pseudosciaena crocea R.) and the histological distribution of OLA and its metabolite 3-methyl-quinoxaline-2-carboxylic acid(MQCA) in fish tissues. Four diets containing 0(control), 42.5, 89.5 and 277.2 mg kg-1 OLA, respectively, were formulated and tested, 3 cages(1.0 m × 1.0 m × 1.5 m) each diet and 100 juveniles(9.75 ± 0.35 g) each cage. The fish were fed to satiation twice a day at 05:00 am and 17:00 pm for 8 weeks. The survival rate of fish fed the diet containing 42.5 and 89.5 mg kg-1 OLA was significantly higher than that of fish fed the diet containing 0 and 277.2 mg kg-1 OLA(P 0.05), while the weight gain rate of fish fed the diet containing 42.5 and 89.5 mg kg-1 OLA was significantly higher than that of fish fed the diet without OLA(control)(P0.05), but similar to that of fish fed the diet with 277.2 mg kg-1 OLA. Fish fed the diet with 277.2 mg kg-1 OLA had the highest content of OLA and MQCA in liver(3.44 and 0.39 mg kg-1, respectively), skin(0.46 and 0.09 mg kg-1, respectively) and muscle(0.24 and 0.06 mg kg-1, respectively). In average, fish fed the diet containing OLA had the highest content of OLA and MQCA in liver which was followed by skin and muscle(P 0.05), whereas OLA and MQCA were not detectable in control. Our findings demonstrated that OLA and MQCA accumulated in large yellow croaker when it was fed with the diet containing OLA, thus imposing a potential safety risk to human health.