The bacterioneuston is defined as the community of bacteria present within the neuston or sea surface microlayer. Bacteria within this layer were sampled using a membrane filter technique and bacterial diversity was compared with that in the underlying pelagic coastal seawater using molecular ecological techniques. 16S rRNA gene libraries of approximately 500 clones were constructed from both bacterioneuston and the pelagic water samples and representative clones from each library were sequenced for comparison of bacterial diversity. The bacterioneuston was found to have a significantly lower bacterial diversity than the pelagic seawater, with only nine clone types (ecotaxa) as opposed to 46 ecotaxa in the pelagic seawater library. Surprisingly, the bacterioneuston clone library was dominated by 16S rRNA gene sequences affiliated to two groups of organisms, Vibrio spp. which accounted for over 68% of clones and Pseudoalteromonas spp. accounting for 21% of the library. The dominance of these two 16S rRNA gene sequence types within the bacterioneuston clone library was confirmed in a subsequent gene probing experiment. 16S rRNA gene probes specific for these groups of bacteria were designed and used to probe new libraries of 1000 clones from both the bacterioneuston and pelagic seawater DNA samples. This revealed that 57% of clones from the bacterioneuston library hybridized to a Vibrio sp.-specific 16S rRNA gene probe and 32% hybridized to a Pseudoalteromonas sp.-specific 16S rRNA gene probe. In contrast, the pelagic seawater library resulted in only 13% and 8% of 16S rRNA gene clones hybridizing to the Vibrio sp. and Pseudoalteromonas sp. probes respectively. Results from this study suggest that the bacterioneuston contains a distinct population of bacteria and warrants further detailed study at the molecular level.
The apparent transfer velocities (kw) of CH4, N2O, and SF6 were determined for gas invasion and evasion in a closed laboratory exchange tank. Tank water (pure Milli‐RO® water or artificial seawater prepared in Milli‐RO®) and/or tank air gas compositions were adjusted, with monitoring of subsequent gas transfer by gas chromatography. Derived kw was converted to “apparent k600,” the value for CO2 in freshwater at 20°C. For CH4, analytical constraints precluded estimating apparent k600 based on tank air measurements. In some experiments we added strains of live methanotrophs. In others we added chemically deactivated methanotrophs, non‐CH4 oxidizers (Vibrio), or bacterially associated surfactants, as controls. For all individual controls, apparent k600 estimated from CH4, N2O, or SF6 was indistinguishable. However, invasive estimates always exceeded evasive estimates, implying some control of gas invasion by bubbles. Estimates of apparent k600 differed significantly between methanotroph strains, possibly reflecting species‐specific surfactant release. For individual strains during gas invasion, apparent k600 estimated from CH4, N2O, or SF6 was indistinguishable, whereas during gas evasion, k600‐CH4 was significantly higher than either k600‐N2O or k600‐SF6, which were identical. Hence evasive k600‐CH4/k600‐SF6 was always significantly above unity, whereas invasive k600‐CH4/k600‐SF6 was not significantly different from unity. Similarly, k600‐CH4/k600‐SF6 for the controls and k600‐N2O/k600‐SF6 for all experiments did not differ significantly from unity. Our results are consistent with active metabolic control of CH4 exchange by added methanotrophs in the tank microlayer, giving enhancements of ∼12 ± 10% for k600‐CH4. Hence reactive trace gas fluxes determined by conventional tracer methods at sea may be in error, prompting a need for detailed study of the role of the sea surface microlayer in gas exchange.
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