Quantification of toxic Microcystis spp. during the 2003 and 2004 blooms in western Lake Erie using quantitative real-time PCR.

In August of 2003 and August of 2004, blooms of potentially toxic cyanobacteria Microcystis spp. persisted in western Lake Erie. Samples collected from the bloom were analyzed for the cyanobacterial toxin microcystin and the presence of Microcystis spp. cells. Estimates of microcystin toxicity exceeding 1 microg L(-1) (microcystin-LR activity equivalents), the safety limit set by the World Health Organization, were found from the samples in both 2003 and 2004. The presence of Microcystis spp. in water samples was confirmed through standard polymerase chain reaction (PCR) using a combination of four primer sets. Quantification of Microcystis was accomplished by a real-time PCR assay utilizing specific primer-Taq-man probe sets targeted on a conserved, Microcystis-specific 16S rDNA fragment and a microcystin toxin synthetase gene mcyD. This approach allowed us to specifically study the distribution and abundance of toxic Microcystis in the lake in contrast to previous studies that have assessed Microcystis populations with less refined methods. On the basis of quantification by quantitative real-time PCR analysis, the total abundance of Microcystis cells in the bloom area varied from 4 x 10(8) to 2 x 10(3) cells L(-1). The results of this study provide novel insight regarding the distribution and abundance of Microcystis spp. in the western basin of Lake Erie, a region plagued in recent years by large-scale (>20 km2) blooms. Our results suggest that the Maumee River and Bay may serve as a source for Microcystis to western and central Lake Erie.

A dilution technique for the direct measurement of viral production: a comparison in stratified and tidally mixed coastal waters.

The abundance of heterotrophic bacteria and viruses, as well as rates of viral production and virus-mediated mortality, were measured in Discovery Passage and the Strait of Georgia (British Columbia, Canada) along a gradient of tidal mixing ranging from well mixed to stratified. The abundances of bacteria and viruses were approximately 10(6) and 10(7) mL(-1), respectively, independent of mixing regime. Viral production estimates, monitored by a dilution technique, demonstrated that new viruses were produced at rates of 10(6) and 10(7) mL(-1)h(-1) across the different mixing regimes. Using an estimated burst size of 50 viruses per lytic event, ca. 19 to 27% of the standing stock of bacteria at the stratified stations and 46 to 137% at the deep-mixed stations were removed by viruses. The results suggest that mixing of stratified waters during tidal exchange enhances virus-mediated bacterial lysis. Consequently, viral lysis recycled a greater proportion of the organic carbon required for bacterial growth under non-steady-state compared to steady-state conditions.

Photoreactivation compensates for UV damage and restores infectivity to natural marine virus communities.

We investigated the potential for photoreactivation to restore infectivity to sunlight-damaged natural viral communities in offshore (chlorophyll a, < 0.1 microgram liter-1), coastal (chlorophyll a, ca. 0.2 microgram liter-1), and estuarine (chlorophyll a, ca. 1 to 5 micrograms liter-1) waters of the Gulf of Mexico. In 67% of samples, the light-dependent repair mechanisms of the bacterium Vibrio natriegens restored infectivity to natural viral communities which could not be repaired by light-independent mechanisms. Similarly, exposure of sunlight-damaged natural viral communities to > 312-nm-wavelength sunlight in the presence of the natural bacterial communities restored infectivity to 21 to 26% of sunlight-damaged viruses in oceanic waters and 41 to 52% of the damaged viruses in coastal and estuarine waters. Wavelengths between 370 and 550 nm were responsible for restoring infectivity to the damaged viruses. These results indicate that light-dependent repair, probably photoreactivation, compensated for a large fraction of sunlight-induced DNA damage in natural viral communities and is potentially essential for the maintenance of high concentrations of viruses in surface waters.

Alterations in cell pigmentation, protein expression, and photosynthetic capacity of the cyanobacterium Oscillatoria tenuis grown under low iron conditions.

To better describe the iron-limited nutrient status of aquatic photosynthetic microorganisms, we examined the effects of iron limitation on pigment content, maximum rates of photosynthetic oxygen evolution, and respiratory oxygen consumption in the filamentous cyanobacterium Oscillatoria tenuis Ag. Within the range of iron (4.2 x 10(-5)-5.1 x 10(-9) M FeCl3), growth rates were not limited by photosynthetic capacity but rather by another, as of yet undetermined, iron-requiring cellular function. We have also investigated membrane proteins that are induced when the cells are grown in low iron medium. Using membrane fractionation techniques we were able to recognize specific proteins localized in the outer membrane and periplasmic space of O. tenuis. The recovery of growth rates at low iron levels occurred in parallel with the induction of these proteins and the production of extracellular siderophores. The additional iron acquired by this high affinity transport system did not reestablish photosynthesis in O. tenuis to the iron-satiated level but did reestablish growth to iron-replete levels. Oscillatoria tenuis appears to invoke an alternate physiology to compensate for iron deficiency.