To optimize spat stocking rates for Pacific calico scallop (PCS, Argopecten ventricosus) during nursery culture, two consecutive 8-week trials were conducted to evaluate growth in land-based downweller and upweller systems. Four stocking rates (60%, 70%, 80%, and 90% of silo bottom coverage) of spat at 3.83 ± 0.10 mm in shell height were tested in the downweller system, whereas three stocking rates (50%, 70%, and 90% of silo bottom coverage) of spat at 0.56 ± 0.029 mm in shell height were evaluated in the upweller system. In the downweller system, spat stocked at 60% generally exhibited the best growth performance, with significantly higher relative growth rate (RGR, mm/day) in shell height compared with higher stocking rates. In the upweller system, spat stocked at 50% and 70% showed similar growth performance and both outperformed the 90% stocking rate treatment in shell height, dry weight, RGR, specific growth rate (SGR, %/day), and condition index (CI). These results suggest that moderate stocking rates, approximately 60% in downweller systems and 50–70% in upweller systems, provide favorable conditions for PCS spat growth while maintaining practical space-use efficiency under the present culture conditions. The present study demonstrates the feasibility of PCS spat nursery culture in land-based systems and provides practical information for hatchery production and nursery management of this emerging aquaculture species.

A tiered flow-through cascade integrated multi-trophic aquaculture (IMTA) system was developed for white seabass (Atractoscion nobilis, WSB), warty sea cucumber (Apostichopus parvimensis, WSC), and seaweeds (Ulva australis and Devaleraea mollis) to enhance resource utilization and reduce nutrient discharge. The system consisted of four tiers of 700 L tanks in each tier. WSB were cultured in first-tier tanks (30 kg/m3), WSC in secondtier tanks (16 ind./tank), and seaweeds in third- and fourth-tier tanks (1 kg/m2 for U. australis, 4 kg/m2 for D. mollis). The seaweed culturing order was switched after 35 days between Phase 1 (P1) and Phase 2 (P2) for a total culture duration of 82 days to compare the performance between different culturing orders. WSB grew at 0.63 f 0.06 and 0.73 f 0.08%/d with a feed conversion ratio of 1.55 f 0.24 and 1.41 f 0.15 in P1 and P2, respectively. WSC removed measurable solid wastes but showed limited growth. Ulva australis grew at 20.22 f 1.71 and 25.90 f 6.72 g DW/m2d with nitrogen uptake rates of 0.92 f 0.08 and 0.90 f 0.12 g/m2d for P1 and P2, respectively, while D. mollis showed lower productivity of 14.66 f 4.54 and 4.41 f 11.57 g DW/m2d resulting in nitrogen uptake rates of 0.58 f 0.10 and 0.23 f 0.59 g/m2d. The nitrogen balance indicated that WSB production accounted for 34.94% of nitrogen output, the "black box" (including N2 production, plumbing retention, incidental autotroph assimilation, and effluent losses) for 30.84%, and seaweeds and WSC harvest for 9.97% and 0.23%, respectively. Leftover nitrogen excreted by animals contributed 10.62%, and measurable leftover WSB wastes contributed 9.74%. Modeling suggested that four additional tanks for each seaweed species and 140 more WSC would be required to fully utilize measurable residual nutrients. Further optimization through improved plumbing design, higher WSC densities, and refined seaweed culture conditions could enhance nutrient utilization efficiency.

Harvesting sea urchins from barrens and enhancing their gonads through aquaculture offers a promising way to convert low-value individuals into high-quality seafood. This study evaluated whether red sea urchins (Mesocentrotus franciscanus, RSUs) fed nutrient-enriched seaweeds produced in a land-based integrated multi-trophic aquaculture (IMTA) system could significantly improve gonad size and quality. Two seaweed species, Ulva australis and Devaleraea mollis, were grown in effluent from white seabass (Atractoscion nobilis) tanks and used to feed RSUs over an 8-week period. RSUs readily consumed both seaweeds, with measurable ingestion, fecal output, and absorption efficiency. We hypothesized that RSUs fed IMTA seaweed would exhibit increased gonadosomatic index and improved gonad quality. GSI significantly increased in both groups, from an initial 3.00 ± 0.50% (9.02 ± 1.80 g) to 4.64 ± 0.66% (23.04 ± 10.20 g) in the U. australis group and to 6.35 ± 1.30% (31.20 ± 7.20 g) in the D. mollis group. Gonad quality improved from “D” (unmarketable) to average “B” (high-quality) grade, based on color, firmness, and texture. These results demonstrate that RSUs collected from barrens can be enhanced into premium seafood using nutrient-enriched seaweeds. Integrating RSUs into land-based IMTA systems may increase aquaculture efficiency, reduce waste, and diversify seafood production in a sustainable and economically viable way.

The warty sea cucumber (Apostichopus parvimensis, WSC) was evaluated for its growth and nutrient assimilation capabilities when fed wastes from white seabass (Atractoscion nobilis, WSB) to assess its potential application in integrated multi-trophic aquaculture (IMTA) systems. Three consecutive trials were conducted: (1) assessment of WSB waste nutritional quality over five days; (2) evaluation of feeding rations at 1 %, 2 %, 3 %, 4 %, and 5 % body wet weight (WW) on 100-150 g wet weight WSCs; and (3) comparison of growth and nutrient assimilation in three size groups of WSCs (small: <100 g, medium: 100-150 g, and large: 151-200 g) fed with 4 % body WW of WSB wastes. All trials were conducted in 175 L tanks with a surface area of 0.89 m(2), run in triplicate, with periodic measurements of leftover WSB wastes, WSC feces, and growth. Results indicated that WSCs effectively assimilated nutrients from WSB wastes, which retained its nutritional value over five days. The 4 % feeding ration was identified as optimal for WSC growth and nutrient assimilation. Additionally, smaller WSCs fed at the 4 % ration exhibited the highest growth rate and nutrient assimilation efficiency. This study demonstrates that WSCs can efficiently utilize WSB wastes as a feed source, achieving relatively high growth rates and nutrient assimilation efficiencies, and highlights their potential in nutrient recycling within WSB IMTA systems.

Warty sea cucumber (Apostichopus parvimensis) is one of two commercial sea cucumber species along the Pacific coast of the U.S., and since 2011, wild populations of A. parvimensis have reached levels of concern due to overfishing. Thus, there is an urgent need to develop A. parvimensis aquaculture to take pressure off wild resources and offer the capability of restoring the depleted stocks through out-planting of cultured animals, while simultaneously supplying data to help improve fishery management. In this study, we present the results on artificial breeding of A. parvimensis, describing the spawning induction, larval development, and the effects of microalgal diets and temperatures on larval performance, aimed at developing a rearing protocol for this species. Wild collected broodstock were induced to spawn by testing four different methods, and peptide (NGLWYamide) injection was proved an effective method for obtaining healthy gametes from this species. Larval development in A. parvimensis progressed through five stages, reaching the pentactula stage in 17 days with a survival rate of 26.67 +/- 2.89 % at 20 degrees C. Five different microalgal combinations were tested as larval feeds at 20 degrees C, and the combination of Rhodomonas sp. and Tisochrysis lutea was found to be the best microalgal diet mix for larval development. In addition to 20 degrees C, larvae survival and development were measured under four other temperature treatments of 22, 24, 26 and 28 degrees C until larvae reached the pentactula stage. We found that 24 degrees C was the upper limit for A. parvimensis larvae to maintain normal development and it also yielded good survival (25.00 +/- 2.50 %). When the temperature reached 26 degrees C and beyond, the larvae stopped developing and shrank in size. This indicated that the temperature for A. parvimensis larvae rearing on farms should be lower than 24 degrees C. Our results indicate that A. parvimensis can be spawned and the larval stages can do well in culture. This will help establish hatchery production protocols and aid in the further development of the aquaculture of this species.

To advance environmentally friendly technologies in the aquaculture of Atractoscion nobilis, and simultaneously to diversify seafood production, a 79-day trial was conducted to assess the performance of Ulva lactuca and Devaleraea mollis cultured in the effluent from A. nobilis in a land-based integrated multi-trophic aquaculture (IMTA) system in southern California, USA. Water quality and performance of macroalgae were measured weekly. The impacted factors on the growth of macroalgae and nutrient uptake rate of macroalgae were assessed. The specific growth rate of juvenile A. nobilis was 0.47–0.52%/d. Total ammonia nitrogen in effluents of A. nobilis tanks ranged from 0.03 to 0.19 mg/L. Ulva lactuca and D. mollis achieved an average productivity of 24.53 and 14.40 g dry weight (DW)/m2/d. The average nitrogen content was 3.48 and 4.89% DW, and accordingly, the average nitrogen uptake rate was 0.88 and 0.71 g/m2/d, respectively. Temperature and nutrient concentration were key factors impacting macroalgae growth, and light intensity also impacted the growth of D. mollis. The high protein content of U. lactuca and D. mollis would make them good for use as human or animal food, or for use in other industries. Research on the interaction effects between seawater exchange rates and aeration rates on the performance and nutrient uptake rates of macroalgae will be conducted in future studies.

A modular land-based integrated multi-tropic recirculating aquaculture system (IMTRAS) for fish, sea cucumbers, mussels and macroalgae was established to optimize resource utilization, while reducing water requirements and nutrient discharge levels. The first module included California yellowtail (Seriola dorsalis) as the primary "fed" species and Ulva lactuca functioned as biofilters. Effluent from the fish tank was passed through a drum filter for solids removal, and then the filtered seawater entered a sump. Seawater was pumped from the sump into U. lactuca tumble culture tanks and then back to the fish tank. The second module housed Mediterranean mussels (Mytilus galloprovincialis) and warty sea cucumbers (Apostichopus parvimensis) in separate serial units. Backwashes of the drum filter occurred automatically and effluent flowed by gravity to M. galloprovincialis culture tanks, and then to A. parvimensis culture troughs. Two trials of two weeks each were conducted - one at an initial fish stocking density of 5.2 kg m(-3) (Trial 1), and another at 20.9 kg m(-3) (Trial 2). The seawater flow rates and aeration levels were determined based on earlier pilot studies. Yellowtail grew on average 0.36 and 0.64% d(-1) with a food conversion ratio of 1.73 and 1.47, in Trial 1 and Trial 2, respectively. Ulva lactuca showed productivity of 24.39 +/- 2.00 and 15.57 +/- 2.47 g DW m(-2) d(-1) with a protein content of 15.13 +/- 3.11 and 30.36 +/- 1.41% DW in each trial, respectively, which resulted in nitrogen uptake rates of 0.58 +/- 0.08 and 0.75 +/- 0.12 g m(-2) d(-1). Mussels and sea cucumbers removed 16.1 to 18.9% and 16.4 to 20.4% of the total solid wastes produced daily, respectively. The condition index of mussels and sea cucumbers showed no difference between the beginning and end of trial. After calculating the nitrogen balance in this IMTRAS, the largest proportion of nitrogen was in the form of dissolved inorganic nitrogen that accumulated inside the system (31.46%), followed by the yield of S. dorsalis (24.64%) and then the harvest production of U. lactuca (14.91%). All other measured nitrogen components were <7 % combined. Nitrogen that was not directly measured (i.e. "black box" nitrogen) including nitrogen accumulated inside plumbing components and lost with the effluent discharge totaled 22.48%. Based on these results, the proper ratio of co-cultured species in this system was estimated as 20.9 kg m(-3) of S. dorsalis, six tanks of U. lactuca, three tanks of M. galloprovincialis and six troughs of A. parvimensis with the same volumes and stocking densities reported in this study. Future long-term trials need to be conducted to refine the operation of this IMTRAS.

Three consecutive 3-week trials were conducted to assess the growth, productivity, and nutrient removal rates of sea lettuce (Ulva lactuca) cultured in effluents from white seabass (Atractoscion nobilis) raceways. All U. lactuca cultivation trials were conducted in 175 L tanks with a surface area of 0.89 m(2) run in triplicate, with water quality and growth measurements taken weekly. For the shading trial, U. lactuca was stocked at 0.56 kg/m(2) with either 0%, 30%, or 60% shade and seawater supplied at 63 tank volumes per day (vol./day) water exchange (7.6 L/min) for each tank. The removal rate of total ammonia nitrogen (TAN) decreased with increasing shade levels, and the peak exceeded 80% removal at 0% shading. Growth rate and productivity of U. lactuca under 0% shading was 12.09 +/- 3.13%/d and 13.28 +/- 4.66 g DW/m(2)/d, which were both significantly higher than other shading levels. For the stocking density and water exchange rate trial, the stocking density of U. lactuca was 0.56 or 1.12 kg/m(2), and seawater exchange rates were 4, 12, or 63 vol./day in a 2-factor design. In this trial, TAN removal rates were 100% across all treatment combinations. Growth rate of U. lactuca was highest (20.36 +/- 2.36%/d) at 0.56 kg/m(2) under the exchange rate of 63 vol./day, and the productivity reached up to 30.89 +/- 6.53 g DW/m(2)/d, which was not different than that of U. lactuca stocked with 1.12 kg/m(2) at the same exchange rate (p = 0.47). In a trial comparing sand-filtered seawater (RAW seawater) with seawater effluent from a raceway containing A. nobilis (FISH seawater), TAN removal rate by U. lactuca was consistently nearly 100% when supplied with FISH seawater with a TAN concentration ranging from 0.11 to 0.18 mg/L. Growth rate and productivity of U. lactuca supplied with FISH seawater were 21.36 +/- 2.25%/d and 33.83 +/- 7.29 g DW/m(2)/d, respectively, which was not different than that for U. lactuca supplied with RAW seawater. However, the protein content of U. lactuca cultured with FISH seawater was significantly higher and C/N ratio significantly lower than those cultured with RAW seawater (p < 0.01), which indicated that U. lactuca was in a nitrogen-limited situation when cultured with RAW seawater. In light of the better understanding of U. lactuca performance achieved in our study, the optimum U. lactuca and A. nobilis density combinations will be confirmed in future studies based on nitrogen balance to maximize the nutrient removal rates and diversity of seafood production.

The goal was to develop a cost-effective and portable freezing system for germplasm cryopreservation of fish and shellfish. The objectives were to: (1) identify components for construction of a portable freezing system by evaluating the affecting factors of cooling rates; (2) generate a range of cooling rates through adjusting different variables; (3) compare cooling profiles generated by the aeration freezing system and a commercial programmable freezer, and (4) verify the effectiveness of the aeration freezing system by cryopreserving sperm of eastern oysters Crassostrea virginica. An aeration freezing system was constructed using a Styrofoam box as cooling chamber and an innovative aeration unit to create nitrogen vapour. A wide range of cooling rates (1.5-32.1 degrees C/min) were achieved by combinations of sample position at 2, 5, or 10 cm above liquid nitrogen surface, the temperature of nitrogen vapour (-150 or -180 degrees C), and aeration time of liquid nitrogen (0, 1, 2 min, or continuity). Compared to a programmable freezer, the aeration freezing system generated cooling profiles with significantly higher temperature for ice nucleation initiation. No difference in post-thaw sperm motility of eastern oysters were found between samples cooling in the aeration freezing system or a programmable freezer. Cooling rates of 10 or 15 degrees C/min from 4 degrees C to -80 degrees C yielded the highest post-thaw sperm motility regardless of the cooling devices. The aeration freezing system developed in this study has the potential for high-throughput sample processing, and the design allows easy construction by multiple users.

The effect of salinity on the survival, growth, development and neonate production of the cladoceran Scapholeberis mucronata (O. F. Müller) was studied by using Chlorella pyrenoidosa as feed at 1, 2, 3, 4 salinity gradients and control group according to the pre-experiment and median lethal concentration experiment. The acute effect of salinity on this species indicated that 4 and 4.5 was its limit superior of reproduction and survival. The survival rate and growth rate of individuals reared at 1 and 2 salinity gradients were higher than that of those reared at the other gradients. The mean size of the female adult decreased from 820 to 743 μm when the salinity increased from 1 to 4. Among individuals reared at 1 and 2, the intrinsic rate of natural increase (rm) of population was 1.021 and 0.903, respectively; the rate of egg production was 1.281 and 1.390, respectively; the cumulative egg production was 83.2 and 106.0 and the mean life span was 16.05 and 17.30, respectively. These values of life history parameters were higher than those of individuals reared at 3. No eggs were produced by females reared at 4 during the whole experiment. Furthermore, individuals reared at 1 and 2 had faster embryonic development. The above results implied that S. mucronata is relatively well adapted to low-salinity conditions (1–2). Resting egg formation and sexual reproduction did not occur at all the tested salinity gradients.

The expansion of the global macroalgal aquaculture and climate change creates the need for germplasm preservation of valuable aquaculture strains and maintenance of natural biodiversity. Compared to the large number of studies in fish and shellfish species, relative few studies have been conducted on the macroalgal germplasm cryopreservation. The first cryopreservation of macroalgae to 75 degrees C was reported on Neopyropia tenera (formerly called Porphyra tenera) in 1964. To date, a total of 34 studies reported germplasm cryopreservation in 33 species, including Chlorophyta (7 species), Ochrophyta (14 species), and Rhodophyta (12 species). The goal of this review was to summarize the published studies on macroalgal germplasm cryopreservation, compare the reported protocols for the cryopreservation process, and identify the factors affecting post-thaw viability. Overall, macroalgal germplasm cryopreservation included haploid or diploid thalli, spores, and gametes. Cryotubes (1.5-ml or 2-ml) have been widely used to package germplasm samples for cooling and storage in most studies, and the 0.5-ml straws and 5-ml cryotubes have been used in several studies. Two approaches (programmable controlled cooling and vitrification) were employed for macroalgal germplasm cryopreservation. A two-step programmable controlled cooling (e.g., from initial culture temperature to a frozen temperature, such as 40 degrees C, and then directly plunging into liquid nitrogen at 196 degrees C) was determined to be an effective cooling strategy. Vitrification, a super rapid cooling for a sample to form non-crystalline amorphous solid, was applied on macroalgal germplasm cryopreservation with sample encapsulation and dehydration. Survival of post-thaw samples varied significantly in different studies. Based on research updates, recommendations are made for future research. It is expected that this review can serve as a foundation for future germplasm banking of macroalgae for aquaculture and biodiversity preservation.

Scyphozoan jellyfish are conspicuous components of marine ecosystems, which during a bloom, can impact food web structure and economically important fisheries. Jellyfish biomass in the southeastern Bering Sea (SEBS) is primarily composed of Chrysaora melanaster and has varied widely over the past four decades, yet the underlying causes of these biomass fluctuations remain unclear. The present study investigated the spatial and seasonal dynamics of C. melanaster along the Alaska Peninsula in the SEBS using an adaptive resolution imaging sonar system and nets in June-July and September 2018. The abundance of C. melanaster was high in coastal waters near the Alaska Peninsula, with peak densities occurring east of Unimak Island in both sampling periods. The current pattern revealed by an acoustic Doppler current profiler showed that cold, deep water from Bering Canyon flowed onto the shelf east of Unimak Pass. Differences in the strength of this cross-isobath, deep-water intrusion were observed during the two sampling periods. This flow may have influenced the distribution of C. melanaster by constraining the mixed coastal water near of Unimak Island in June-July and by transporting medusae westward, into deeper water in September, after the deep-water intrusion weakened. The present study provides a new mechanistic understanding of how regional-scale ocean variability in the SEBS affects C. melanaster distribution.

The Eastern oyster Crassostrea virginica (Family Ostreidae) is one of the most important fishery and aquaculture species in the U.S. and is a keystone species for coastal reefs. A breeding program was initiated in 2019 to support the fast-growing aquaculture industry culturing this species in the Gulf of Mexico. Oysters from 17 wild populations in embayment along the U.S. Gulf of Mexico coast from southwest Florida to the Matagorda Bay, Texas were used as broodstock for the program to maximize genetic diversity in the base population. A sperm repository of the broodstock was established to support the breeding project. The goal of this study was to demonstrate the sperm sample collection, processing, cryopreservation, and the data management plan involved in the establishment of a sperm germplasm repository of base populations. The supporting objectives were to: (1) develop a data management plan for the sperm repository; (2) streamline the procedure for sample collection, processing, and cryopreservation; (3) incorporate sperm quality analysis into the procedure, and (4) archive the cryopreserved samples as a repository for future use in the breeding program. This sperm repository included a total of 102 male oysters from the 17 collection sites (six oysters per site). A data management plan was developed with six categories, including sample collection, phenotype, fresh sperm, genotype, cryopreservation, and post-thaw sperm, as guide for data collection. Sperm collection was accomplished by strip spawn, and fresh sperm production, motility, and fertility were recorded for quality analysis. Cryopreserved sperm samples were sorted, labelled, archived, and stored in liquid nitrogen for future use. Post-thaw motility (1–30%) and plasm membrane integrity (15.34–70.36%) were recorded as post-thaw quality parameters. Overall, this study demonstrated a streamlined procedure of oyster sperm collection, processing, and cryopreservation for establishing a sperm repository that can serve as a template for construction of oyster germplasm repositories for breeding programs.

The northern quahog Mercenaria mercenaria (commonly named hard clam) is an important aquaculture and fishery species along the Atlantic west coast. Environmental stresses, such as heat shock, fluctuating salinity, and harmful algal blooms are major challenges for clam aquaculture. In response to environmental stresses, hemocytes would change dynamically for defense and immunity. The goal of this study was to characterize basic immunological assays of hemocytes in the northern quahog by use of flow cytometry. The objectives were to: 1) develop a non-lethal method for hemolymph collection and dilution; 2) verify the capability of flow cytometry for hemocyte count and type identification through comparison with microscopic observation; 3) validate hemocyte viability assay based on plasma membrane integrity, and 4) develop hemocyte phagocytosis assay by use of fluorescein labeled microbeads. A non-lethal hemocyte collection method was developed using needle insertion through the ligament. Osmolality measurement of serum was the same as that of culture seawater. The pH measurement of serum (7.2) was significantly different from that of culture seawater (8.4). By microscopic observation, three types of hemocytes were identified with granulocytes, the dominant cell type (70 ± 16%), agranulocyte (14 ± 4%), and blast-like cell (16 ± 4%), and no differences were found from the measurements by flow cytometer on FSC/SSC plot (cell size/granularity). The viability of hemocytes based on plasma membrane integrity was 88 ± 6% ranging from 70 to 97% (n = 60, three populations), and viability protocol was further validated with the pre-set expected viability (p ≥ 0.424). Phagocytosis assay of hemocytes with fluorescence beads showed a mean capacity of 10 ± 5% (n = 60, three populations). Incubation time (up to 6 h) or bead concentrations (2:1 or 5:1 to hemocytes) did not affect the phagocytosis measurement. Overall, this study reported the basic characteristics of hemolymph (serum and hemocytes) of northern quahogs. It is expected that the assay methodologies will be applied to evaluation of hemocyte responses to environmental stresses for clam aquaculture.

This review is focused on the status and summary of larval cryopreservation in aquaculture mollusks. A total of 26 publications were identified addressing 14 molluscan bivalve species. Most studies were conducted on trochophores and D-larvae, a few studies were on umbo larvae, and no study was found on pediveligers. Based on the post-thaw viability, there is no general conclusion about the best larval stage for cryopreservation. The research topics of most publications focused on developing or improving cryopreservation protocols through exploring one or more vital factors of the cryopreservation procedure. The most dominant cryoprotectant agents (CPAs) used for molluscan larval cryopreservation were dimethyl sulfoxide (DMSO), propylene glycol (PG), and ethylene glycol (EG) at final concentrations of 5-15%. Non-permeable CPAs, such as glucose, fructose, sucrose, trehalose, and polyvinylpyrrolidone (PVP), were dominantly used as additives. Most cooling processes started from 0 degrees C, 4 degrees C, or larval culture temperature to -10 degrees C or - 12 degrees C at -1 degrees C/ min and holding at this temperature for 5-15 min with or without "seeding", and then continued to cool to around -35 degrees C at -0.3 to -2.5 degrees C/ min (mostly at -0.5 and - 1 degrees C/ min) followed by a 5-10 min holding period. All frozen samples were then transferred into liquid nitrogen (-196 degrees C) for long-term storage. Evaluation of post-thaw viability was reported over a wide range of parameters with no standard criteria, for example, motile post-thaw larvae were considered alive but no definition of swimming velocity. A sharp decrease of survival was often reported after a period of culture although immediate post-thaw survivals were reported high in several studies. Overall, no general optimal protocol could be concluded. Post-thaw larvae were reported to survive beyond metamorphosis in Pacific oysters and even to the adult stage and reproduced successfully after 3 years of culture, indicating the promising future of this technology. Till now, no study has been reported on germplasm cryobanking by use of molluscan larval cryopreservation. Pathways and aspects for promoting molluscan larval cryopreservation technology for germplasm repository establishment and commercial seed production were discussed.

目的:制备三氧化二砷纳米粒并进行细胞学实验,分析体外药效及其对耐药细胞的逆转倍数.方法:运用CCK-8法考察纳米粒的细胞毒性以及纳米粒对乳腺癌细胞MCF-7/ADR多药耐药的逆转;采用Annexin V-FITC/PI双染法,用流式细胞仪检测三氧化二砷纳米粒诱导细胞凋亡的作用;采用PI单染流式细胞术检测As2 O3-NPs对MCF-7/ADR周期的阻滞.结果:细胞学实验结果表明,各组药物均能够抑制耐药细胞的正常生长,凋亡率呈浓度依赖性,纳米粒组的凋亡率高于游离药物组;游离药物组和纳米粒组均阻滞细胞生长于G2/M期,As2 O3-mPEG-PLA-NPs阻滞效果最好.As2 O3、As2 O3-PLA-NPs、As2 O3-mPEG-PLA-NPs作为逆转剂时,ADM对MCF-7/ADR的逆转倍数分别为1.32、1.8和2.11.结论:As2 O3-PLA-NPs、As2 O3-mPEG-PLA-NPs均对乳腺癌耐药细胞MCF-7/ADR有促凋亡作用,对G2/M期细胞生长阻滞明显,对MCF-7/ADR的逆转作用强于As2 O3.

目的:制备As2O3纳米粒(As2O3-PLA-NPs、As2O3-mPEG-PLA-NPs),并考察其在大鼠体内吸收生物利用度.方法:采用复乳溶剂挥发法制备三氧化二砷纳米粒(As2 O3-PLA-NPs、As2 O3-mPEG-PLA-NPs),表征其外观形态、粒径、Zeta电位,并检测包封率、载药量及体外释放度.SD大鼠分别尾静脉注射As2 O3生理盐水溶液和As2 O3-PLA-NPs、AS2 O3-mPEG-PLA-NPs注射液,比较药动学行为及生物利用度.结果:As2 O3-PLA-NPs和As2 O3-mPEG-PLA-NPs外观均呈球形,大小相似,分散性良好;平均粒径分别为(133.20±5)nm、(123.40±6)nm,Zeta电位分别为-6.7 mV、-7.11 mV,包封率分别为(86.32±0.54)％、(91.75±0.38)％;两种纳米粒前2 h无突释,均具有一定缓释作用,在72 h累积释放百分率分别为(64.84±1.18)％和(69.15±1.43)％;As2 O3-PLA-NPs组T1/2为As2 O3生理盐水组的1.52倍,AUC为1.42倍,As2 O3-mPEG-PLA-NPs体内长循环效果更显著,T1/2为As2 O3生理盐水组的2.53倍,AUC为2.20倍;mPEG修饰纳米粒子组的T1/2相比于PLA纳米粒组,有显著提高(P<0.05).结论:应用复乳溶剂挥发法成功制备As2 O3-PLA-NPs、As2 O3-mPEG-PLA-NPs,方法简单可行;mPEG修饰的纳米粒子在体外释放,体内药动学方面表现出更好的缓释效果.

目的 探讨柿叶黄酮纳米脂质体的制备工艺,对其进行包衣并表征.方法 采用薄膜分散法-孵育法制备卡波姆包衣的柿叶黄酮纳米脂质体,在单因素考察的基础上,以包封率为评价指标,通过正交设计优化得到最佳制备工艺,检测Zeta电位和粒径;透射电镜下观察形态.结果 主药用量为10mg,药脂比1:20,卵磷脂:胆固醇(PC:Ch)=10:1,磷脂浓度(CPC)为15mg·mL-1,包衣材料浓度5g·L-1,透射电镜下脂质体呈现椭球形囊泡结构,外观均匀圆整.平均Zeta电位为(-47.1±2.2)mV,平均粒径为(257.2±9.3)nm.结论 该处方工艺适用于口服包衣柿叶黄酮纳米脂质体的制备.

The effects of various concentrations of four types of plant growth regulator (PGR) on the physiological performance and microbial populations of three calcifying Halimeda species were evaluated in a 42-day aquaria experiment. The results show that PGRs enhanced the growth and calcification rates at relatively low concentrations, while inhibiting the physiological activities of Halimeda at high concentrations. Net oxygen production rate and pigment concentrations were differed depending on the concentrations and types of PGRs. Carbonic anhydrase activity, total tissue carbon and nitrogen contents varied among different PGRs concentrations, which suggested that these physiological performances were enhanced under appropriate concentration availability of PGRs. Based on 16S rRNA high-throughput sequencing analyses, Alphaproteobacteria, Gammaproteobacteria, and Deltaproteobacteria, belonging to the Proteobacteria phylum, were significantly higher in the PGRs treatment than those in control group. The microbial communities could be majorly divided into three groups corresponding to the Halimeda species, suggesting that physiological differences between Halimeda species may lead to secretion of specific types of biochemical compounds, thus selecting different microbial species at both the taxonomic level and functional level, which may contribute to carbon and nitrogen fixation. The results of this study indicated that it is both practical and efficient to amplify calcifying Halimeda biomass through the addition of PGRs, which could provide a theoretical basis for use of this technique in restoring and maintaining the ecological diversity and sustainable development of coral reef ecosystems.