Relationship between bacteria and phytoplankton during the giant jellyfish Nemopilema nomurai bloom in an oligotrophic temperate marine ecosystem

Bacterial abundance, phytoplankton community structure and environmental parameters were investigated to study the relationships between bacteria and phytoplankton during giant jellyfish Nemopilema nomurai blooms in the central Yellow Sea during 2013. N. nomurai appeared in June, increased in August, reached a peak and began to degrade in September 2013. Results showed that phosphate was possible a key nutrient for both phytoplankton and bacteria in June, but it changed to nitrate in August and September. Phytoplankton composition significantly changed that pico-phytoplankton relative biomass significantly increased, whereas other size phytoplankton significantly decreased during jellyfish bloom. In June, a significantly positive correlation was observed between chlorophyll a concentration and bacterial abundance（r=0.67, P0.001, n=34）.During jellyfish outbreak in August, there was no significant correlation between phytoplankton and bacteria（r=0.11, P0.05, n=25）, but the relationship（r=0.71, P0.001, n=31） was rebuilt with jellyfish degradation in September. In August, small size phytoplankton occupied the mixed layer in offshore stations, while bacteria almost distributed evenly in vertical. Chlorophyll a concentration significantly increased from（0.42±0.056） μg/L in June to（0.74±0.174） μg/L in August, while bacterial abundance just slightly increased. Additionally, the negative net community production indicated that community respiration was not entirely determined by the local primary productivity in August. These results indicated that jellyfish blooms potentially affect coupling of phytoplankton and bacteria in marine ecosystems.

Intraguild predation by polyps of three scyphozoan jellyfish:Nemopilema nomurai,Aurelia coerulea,and Rhopilema esculentum

Jellyfish blooms occur worldwide and have resulted in serious problems in tourism,fisheries,coastal industries,and the marine ecosystem.The life cycle of scyphozoan jellyfish consists of a pelagic medusa stage and a benthic polyp stage.Success of asexual reproduction of the polyps determines directly the number of medusae;thus,the polyp stage is the key to understanding the population dynamics of medusae.Nemopilema nomurai,Aurelia coerulea,and Rhopilema esculentum are three scyphozoan jellyfish commonly inhabit in Chinese coastal waters.Polyps of A.coerulea are easily visible,while those of N.nomurai and R.esculentum remain yet to be found in the wild.However,distribution of the medusa indicates that the polyps of all three species may occur together.To evaluate the distribution pattern of polyps of the three species and explore intraguild predation by the polyps,we conducted a laboratory experiment that considered the attachment sequence and size relationship of calyx diameter of the polyps.We found that the polyps of A.coerulea preyed on polyps of the other two species in all treatments,except when polyps of R.esculentum were bigger than those of A.coerulea.The polyps of R.esculentum preyed on the polyps of N.nomurai only when polyps of R.esculentum attached first and were bigger than those of N.nomurai.Colonies of N.nomurai polyps were rarely found in the places inhabited by A.coerulea polyps.In addition,A.coerulea polyps are known to inhabit at depths of less than 20 m in coastal sea,thus,we speculate that N.nomurai polyp colonies might occur at depths of more than 20 m.Therefore,our finding that polyps of A.coerulea aggressively preyed on polyps of other species may help understand other such systems of jellyfish bloom in the world.

Development of a real-time PCR assay(SYBR Green I) for rapid identification and quantification of scyphomedusae Aurelia sp.1 planulae

The complicated life cycle ofAurelia spp., comprising benthic asexually-reproducing polyps and sexually-reproducing medusae, makes it hard for researchers to identify and track them, especially for early stage individuals, such as planulae. To solve this problem, we developed a real-time PCR assay （SYBR Green I） to identify planulae in both cultured and natural seawater samples. Species-specific primers targeting Aurelia sp.1 mitochondrial 16S rDNA （mr 16S rDNA） regions were designed. Using a calibration curve constructed with plasmids containing the Aurelia sp. 1 mt 16S rDNA fragment and a standard curve for planulae, the absolute number of mt 16S rDNA copies per planula was determined and from that the total number ofplanulae per sample was estimated. For the field samples, a 100-fold dilution of the sample DNA combined with a final concentration of 0.2 μg/μL BSA in the PCR reaction mixture was used to remove real- time PCR inhibitors. Samples collected in Jiaozhou Bay from July to September 2012 were subsequently analyzed using this assay. Peak Aurelia sp.1 planula abundance occurred in July 2012 at stations near Hongdao Island and Qingdao offshore; abundances were very low in August and September. The real-time PCR assay （SYBR Green I） developed here negates the need for traditional microscopic identification, which is laborious and time-consuming, and can detect and quantify jellyfish planulae in field plankton samples rapidly and specifically.

Species-specific detection and quantification of scyphomedusae in Jiaozhou Bay,China,using a quantitative real-time PCR assay

Over the past decades,jellyfish occurred increasingly and abundantly in coastal areas worldwide.Usually,biomass of jellyfish,especially when they bloom,can be determined by visual counting.However,tiny individuals of jellyfish(e.g.,planulae,polyps,and ephyrae)are difficult to detect in the field.In this study,species-specific quantitative real-time PCR assays(qPCR)(SYBR Green I)targeting the mitochondrial 16S rDNA(mt-16S rDNA)of jellyfish were developed and were used to estimate the distribution and seasonal fluctuations of four jellyfish species(Nemopilema nomurai,Cyanea nozakii,Rhopilema esculentum,and Aurelia coerulea)in Jiaozhou Bay(JZB),China in 2013.The mt-16S rDNA of A.coerulea and N.nomurai was detected in most of the survey months and it peaked in July(1.03×10^(4) copies/L)and September(1.08×10^(6) copies/L),respectively.The mt-16S rDNA of C.nozakii occurred from August to October only with monthly mean values of 7.18-46.17 copies/L and was mainly located from the middle part to the outer part of the bay.The mt-16S rDNA of R.esculentum was the least abundant among the four species and was detected in only one sample(B2 station in March),with a value of 88.49 copies/L.The Spearman correlation test revealed that phytoplankton biomass was significantly and positively correlated with the mt-16S rDNA abundance of A.coerulea(R=0.37,P<0.01)and negatively with the mt-16S rDNA of N.nomurai(R=-0.36,P<0.01).The qPCR assay will enable the identification and quantification of jellyfish species in their whole life history and can be used as an approach in combination of the traditional jellyfish survey.

Experimental clearance rates of Aurelia coerulea ephyrae and medusae,and the predation impact on zooplankton in Jiaozhou Bay

The population explosion of jellyfish Aurelia coerulea occurred in Jiaozhou Bay,China in 2009.The potential predation impact of A.coerulea on zooplankton was investigated.Population clearance potential and residence time(t 1/2)for copepods were calculated from laboratory clearance rates and measurements of jellyfish size and abundance from May to August 2009 in Jiaozhou Bay.Clearance rates varied widely with prey organisms,but they were not significantly different among various prey concentrations.Medusae captured rotifers,fish larvae and hydromedusae more efficiently than fish eggs,copepods and chaetognaths.Ephyrae captured rotifers and hydromedusae more efficiently than fish larvae and copepods.Clearance rate linearly increased with the cross sectional area of A.coerulea(size from 0.3 to 7.1 cm).Water temperature also had a marked effect on clearance rate and this was related to the increased beat frequency as water temperature increased.In early May 2009,A.coerulea potentially cleared the volume of water in the Bay less than 0.001 times a day,but this value was estimated to be more than 0.3 times a day in July.The t 1/2 for copepods was less than 6 d in June and July.Abundances of copepods,hydromedusae and chaetognaths were extremely low in 2009 compared to 2008 and 2010(jellyfish non-bloom years).Large predation pressure by the A.coerulea population occurred to control zooplankton communities in Jiaozhou Bay.A.coerulea,when present at a high population level,can be a keystone species in Jiaozhou Bay and control the trophic structure here.

Natural predators of polyps of three scyphozoans:Nemopilema nomurai,Aurelia coerulea,and Rhopilema esculentum

Jellyfish blooms have become a hot research topic in recent decades because they pose a serious threat to fisheries,coastal industries,tourism,and the marine ecosystem.The life cycle of scyphozoan jellyfish consists of a pelagic medusa stage and a benthic polyp stage,where asexual reproduction and strobilation of the polyps directly affect the abundance of ephyra and subsequently medusa abundance.The dynamics of polyps are affected by both environmental and biological factors,and predation by natural predators is one of the most important biological factors.Nemopilema nomurai,Aurelia coerulea,and Rhopilema esculentum are three scyphozoan species that are commonly found in Chinese coastal waters,and previous studies reported that the survivorship of polyps differs among the three species when they are exposed to the same benthic community.To identify potential natural predators of polyps of these three species in Chinese coastal waters and to determine whether the predation rates on polyps of the three species differ,we collected 39 species of macrozoobenthos from the Bohai Sea,Yellow Sea,and East China Sea from May 2014 to June 2016 and conducted predation tests and predation rate measurements.We found that the nudibranchs Pleurobranchaea novaezealandiae,Okenia plana,and Chromodoris tinctoria and the sea anemones Paracalliactis sinica,Calliactis japonica,Anthopleura incerta,and Anthopleura midori could prey on the polyps of all three scyphozoan species.The predation rates increased with the body length of the predators.The predations rates were also related to the polyp species,although the different predators showed no consistent preference for a particular species of polyp.Our results indicate that introducing predators to locations inhabited by polyps might be a way to control the benthic polyp populations and prevent subsequent jellyfish blooms.