Flow cytometric determination of bacterial abundance in lake plankton with the green nucleic acid stain SYTO 13

Giorgio, Paul A. del; Bird, David F.; Prairie, Yves T.; Planas, Dolors

doi:10.4319/lo.1996.41.4.0783

Cited by 309 publications

(255 citation statements)

References 6 publications

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“…Samples for bacterial enumeration were preserved in glutaraldehyde and subsequently analyzed using a FACS caliber Becton Dickson flow cytometer according to the method of del Giorgio et al (1996). A 100 ml sample aliquot was also preserved using Lugol's preservative for phytoplankton community composition and enumeration.…”

Section: Gulf Of Mexicomentioning

confidence: 99%

“…Both sets of samples were frozen over desiccant and analyzed within 1 wk by the fluorometric method of Holm-Hansen et al (1965). An additional GF/F filter of the unfractionated particulates was stored on liquid N 2 for subsequent analysis of phytoplankton pigments using a Hewlett Packard high-performance liquid chromatograph (Model 1100) system, according to the method of Van Heukelem et al (1994) and Van Heukelem & Thomas (2001).Samples for bacterial enumeration were preserved in glutaraldehyde and subsequently analyzed using a FACS caliber Becton Dickson flow cytometer according to the method of del Giorgio et al (1996). A 100 ml sample aliquot was also preserved using Lugol's preservative for phytoplankton community composition and enumeration.…”

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confidence: 99%

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Evidence for dissolved organic nitrogen and phosphorus uptake during a cyanobacterial bloom in Florida Bay

Glibert¹,

Heil²,

Hollander³

et al. 2004

Mar. Ecol. Prog. Ser.

169

121

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Florida Bay, a shallow, seagrass-dominated bay in southern Florida, USA, receives significant nutrient inputs and has experienced seagrass losses and microalgal blooms within the last several decades. Inorganic nutrient inputs have been well characterized, but the role of organic nutrients, specifically of dissolved organic nitrogen (DON) and organic phosphorus (DOP), in supporting microbial processes in the bay is unknown. In this study various techniques were used to assess the importance of these nutrients along a transect in Florida Bay when a cyanobacterial bloom occurred in the central region in November 2002. These techniques included measurements of ambient particulate and dissolved nutrients, enzyme (urease and alkaline phosphatase) activities, and experiments to determine rates of 15 N uptake (nitrate, ammonium, urea, and amino acids over a period of 0.5 h) and long-term (48 h) changes in microbial biomass and 15 N natural abundance in enrichment bioassays. The cyanobacterial bloom in central Florida Bay was associated with the highest concentrations of DON and DOP, whereas the microflagellate-and diatom-dominated eastern bay region was associated with the highest concentrations of inorganic nutrients. The zeaxanthin:chlorophyll a ratio (an indicator of the relative contribution of cyanobacteria to phytoplankton biomass) was positively correlated with the rate of uptake of urea, and negatively correlated with the rate of uptake of inorganic nitrogen. The opposite pattern was observed for the fucoxanthin:chlorophyll a ratio (indicative of relative diatom biomass) and the peridinin:chlorophyll a ratio (indicative of relative photosynthetic dinoflagellate biomass), suggesting that different algal groups were using different N substrates. Biomass responses in the bioassay experiments showed that phytoplankton (as chlorophyll a) responded to DON additions in the western region and to DOP additions in the eastern region, but heterotrophic bacteria, in contrast, responded to DOP additions in the west and DON additions in the east. These findings thus demonstrate the potential for different sources of N, including DON, to stimulate different components of the algal community, and for the phytoplankton and bacteria to respond differently to N and P.KEY WORDS: Florida Bay · Cyanobacteria · Organic N · Organic P · Nutrient limitation · Urease · Alkaline phosphatase · Bioassay · Nutrient uptake rates Resale or republication not permitted without written consent of the publisherMar Ecol Prog Ser 280: [73][74][75][76][77][78][79][80][81][82][83] 2004 changes. Since the onset of industrialization in the 1880s, the health of the Florida Bay ecosystem has been negatively impacted on both decadal (e.g. increasing eutrophication) and centurial (e.g. changes in land use and water management practices within southern Florida) time-scales (Fourqurean & Robblee 1999). These anthropogenic changes have led to a significant alteration in freshwater flow patterns within the Everglades, causing declines in seagrass d...

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Section: Gulf Of Mexicomentioning

confidence: 99%

mentioning

confidence: 99%

Evidence for dissolved organic nitrogen and phosphorus uptake during a cyanobacterial bloom in Florida Bay

Glibert¹,

Heil²,

Hollander³

et al. 2004

Mar. Ecol. Prog. Ser.

169

121

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“…Prokaryotic and viral abundance were determined by flow cytometry (Del Giorgio et al, 1996;Brussaard, 2004). For enumerating prokaryotes, 2 ml samples were fixed with glutaraldehyde (0.5% final concentration), shock-frozen in liquid N 2 and kept at À80 1C until analysis.…”

Section: Prokaryotic and Viral Abundancementioning

confidence: 99%

Links between viral and prokaryotic communities throughout the water column in the (sub)tropical Atlantic Ocean

et al. 2010

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Viral and prokaryotic abundance, production and diversity were determined throughout the water column of the subtropical Atlantic Ocean to assess potential variations in the relation between viruses and prokaryotes. Prokaryotic abundance and heterotrophic activity decreased by one and three orders of magnitude, respectively, from the epi-to the abyssopelagic layer. Although the lytic viral production (VP) decreased with depth, lysogenic VP was variable throughout the water column and did not show any trend with depth. The bacterial, archaeal and viral community composition were depth-stratified as determined by the automated ribosomal intergenic spacer analysis, terminalrestriction fragment length polymorphism and randomly amplified polymorphic DNA-PCR, respectively. Generally, the number of operational taxonomic units (OTUs) did not reveal consistent trends throughout the water column. Viral and prokaryotic abundance were strongly related to heterotrophic prokaryotic production, suggesting similar linkage strength between the viral and prokaryotic communities from the lower epi-to the abyssopelagic layer in the Atlantic Ocean. Strikingly, the prokaryotic and viral parameters exhibited a similar variability throughout the water column down to the abyssopelagic layers, suggesting that the dark ocean is as dynamic a system as is the lower epipelagic layer. It also indicates that viruses are apparently having a similar role for prokaryotic mortality in the dark oceanic realm as in surface waters. The more than twofold increase in bacterial OTUs from 2750 m depth to 45000 m depth and the concurrent decrease in viral OTUs, however, suggests that viruses might exhibit a wider host range in deep waters than in surface waters.

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“…Unpigmented cells can be stained with SYTO-13 (Invitrogen, Carlsbad, CA) (Del Giorgio, et al, 1996;Stepanauskas & Sieracki, 2007) and 4',6-diamidino-2-phenylindole (DAPI) and detected 4 using the 488 nm or UV laser (355 nm) (JDS Uniphase Xcyte Laser), respectively. The multiple lasers of the Influx™ are especially useful when cells are hybridized to synthetic or antibodybased probes.…”

Section: Flow Cytometer Featuresmentioning

confidence: 99%

Coupling FACS and Genomic Methods for the Characterization of Uncultivated Symbionts

Thompson

Bench

Carter

et al. 2013

Methods in Enzymology

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Symbioses between microbes are likely widespread and functionally relevant in diverse biological systems however they are difficult to discover. Most microbes remain uncultivated, symbioses can be relatively rare or dynamic, and intercellular connections can be delicate. Thus traditional methods such as microscopy are inadequate for efficient discovery and precise characterization of novel interactions, their metabolic basis, and the species involved. Highthroughput metagenomic sequencing of entire microbial communities has revolutionized the field of microbial ecology, however genomic signals from symbionts can get buried in sequences from abundant organisms and evidence for direct links between microbial species cannot be gained from bulk samples. Thus, a specialized approach to the characterization of symbioses between naturally-occurring microbes is required. This chapter presents methods for combining fluorescence-activated cell sorting (FACS) to isolate and separate uncultivated symbionts with molecular biology techniques for DNA amplification in order to characterize uncultivated symbionts through genomic and metagenomic techniques.

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Flow cytometric determination of bacterial abundance in lake plankton with the green nucleic acid stain SYTO 13

Cited by 309 publications

References 6 publications

Evidence for dissolved organic nitrogen and phosphorus uptake during a cyanobacterial bloom in Florida Bay

Evidence for dissolved organic nitrogen and phosphorus uptake during a cyanobacterial bloom in Florida Bay

Links between viral and prokaryotic communities throughout the water column in the (sub)tropical Atlantic Ocean

Coupling FACS and Genomic Methods for the Characterization of Uncultivated Symbionts

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