Comparison of the Pour, Spread, and Drop Plate Methods for Enumeration of
            <i>Rhizobium</i>
            spp. in Inoculants Made from Presterilized Peat

Hoben, Heinz J.; Somasegaran, Padma

doi:10.1128/aem.44.5.1246-1247.1982

Cited by 318 publications

(148 citation statements)

References 2 publications

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“…Strains were stored with TSB-1 plus dimethylsulfoxide 10% at -80°C. All bacterial counts were made on TSA-1 or MSWYE-1 agar by the drop plate methodology of Hoben and Somasegaran (1982).…”

Section: Plasmids Bacterial Strains and Growth Conditionmentioning

confidence: 99%

Role of the metalloprotease Vvp and the virulence plasmid pR99 of Vibrio vulnificus serovar E in surface colonization and fish virulence

Valiente¹,

Lee²,

Hor³

et al. 2007

Environmental Microbiology

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The virulence for eels of Vibrio vulnificus biotype 2 serovar E (VSE) is conferred by a plasmid that codifies ability to survive in eel serum and cause septicaemia. To find out whether the plasmid and the selected chromosomal gene vvp plays a role in the initial steps of infection, the VSE strain CECT4999, the cured strain CT218 and the Vvp-deficient mutant CT201 (obtained in this work by allelic exchange) were used in colonization and virulence experiments. The eel avirulent biotype 1 (BT1) strain YJ016, whose genome has been sequenced, was used for comparative purposes. The global results demonstrate that the plasmid does not play a significant role in surface colonization because (i) CECT4999 and CT218 were equally chemoattracted towards and adherent to eel mucus and gills, and (ii) CT218 persisted in gills from bath-infected eels 2 weeks post infection. In contrast, mutation in vvp gene reduced significantly chemoattraction and attachment to eel mucus and gills, as well as virulence degree by immersion challenge. Co-infection experiments by bath with CECT4999 and CT201 confirmed that Vvp was involved in eel colonization and persistence in gills, because CECT4999 was recovered at higher numbers compared with CT201 from both internal organs of moribund fish (ratio 4:1) and gills from survivors (ratio 50:1). Interestingly, YJ016 also showed chemoattraction and attachment to mucus, and complementation of CT201 with BT1-vvp gene restored both activities together with virulence degree by immersion challenge. Additional experiments with algae mucus and purified mucin gave similar results. In conclusion, the protease Vvp of V. vulnificus seems to play an essential role in colonization of mucosal surfaces present in aquatic environments. Among the V. vulnificus strains colonizing fish mucus, only those harbouring the plasmid could survive in blood and cause septicaemia.

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“…Strains were stored with TSB-1 plus dimethylsulfoxide 10% at -80°C. All bacterial counts were made on TSA-1 or MSWYE-1 agar by the drop plate methodology of Hoben and Somasegaran (1982).…”

Section: Plasmids Bacterial Strains and Growth Conditionmentioning

confidence: 99%

Role of the metalloprotease Vvp and the virulence plasmid pR99 of Vibrio vulnificus serovar E in surface colonization and fish virulence

Valiente¹,

Lee²,

Hor³

et al. 2007

Environmental Microbiology

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show abstract

“…Viable counts were measured by colony forming units (CFU) on VNSS agar [16] using the drop plate method [17]. The optical density (0D610) was monitored, using a spectrophotometer (Novaspec, LKB) at each sampling time during 120 h of starvation.…”

Section: Vtability Measurementsmentioning

confidence: 99%

Changes in viability, respiratory activity and morphology of the marine Vibrio sp. strain S14 during starvation of individual nutrients and subsequent recovery

Holmquist

Kjelleberg

1993

FEMS Microbiology Ecology

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Starvation for different individual nutrients revealed various morphotypes of Vibrio sp. strain S14. Carbon or multiple‐nutrient starved cells formed ultramicrocells with low respiratory activity and high culturability. In contrast, cells starved for nitrogen or phosphorus formed filaments or swollen rods with large inclusion bodies of PHB. These cells exhibited a 3–4 log decrease in culturability, nevertheless many were actively respiring and direct viable counts equalled at least 80% of the original number of cells. After 2–3 days of prolonged incubation, microcolonies appeared at approximately the same number of cells as at the onset of starvation. A nutrient‐induced increase in respiratory activity, after 120 h of starvation, was instantaneous for cells starved for carbon or multiple‐nutrients, but cells depleted of nitrogen or phosphorus exhibited a lag period of at least 3 h.

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“…Three manual plating methods (Table 6) were evaluated for the enumeration of Rhizobium spp. in peat inoculants [57]. Spread-plate, pour-plate and drop-plate are the more commonly used methods for plating a recovery solution.…”

Section: Isolation Methodsmentioning

confidence: 99%

A review of issues related to the quantification of bacteria from the phyllosphere

Jacques¹,

Morris²

2006

FEMS Microbiology Ecology

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Quantification of the size of epiphytic bacterial populations and characterization of their composition involves definition of a sampling strategy in time and space, the choice of methods for liberating the bacteria from the leaf surface and for recovering them for subsequent determination of the number of viable or culturable cells. This literature review focuses on some of the issues related to these choices. After briefly reviewing the different types of epiphytic colonizers we consider the biological, methodological, and statistical consequences of the choice of the sampling unit and of the spatial and temporal variability of population size and composition for epiphytic bacteria. The different methods available for the detection and enumeration of naturally occurring microorganisms in the phyllosphere are discussed. Advantages and drawbacks of each are described in this review designed as a ‘hands‐on’ guide.

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Comparison of the Pour, Spread, and Drop Plate Methods for Enumeration of Rhizobium spp. in Inoculants Made from Presterilized Peat

Abstract: Inoculants prepared with presterilized peat were enumerated by the pour, spread, and drop plate techniques. Results indicated that the three plating methods were interchangeable. The drop plate method was preferred because of its economy in materials and labor.

Cited by 318 publications

References 2 publications

Role of the metalloprotease Vvp and the virulence plasmid pR99 of Vibrio vulnificus serovar E in surface colonization and fish virulence

Role of the metalloprotease Vvp and the virulence plasmid pR99 of Vibrio vulnificus serovar E in surface colonization and fish virulence

Changes in viability, respiratory activity and morphology of the marine Vibrio sp. strain S14 during starvation of individual nutrients and subsequent recovery

A review of issues related to the quantification of bacteria from the phyllosphere

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