Inter-laboratory validation of the serial dilution culture—most probable number method for enumerating viable phytoplankton

MacIntyre, Hugh L.; Cullen, John J.; Rastin, Shannah; Waclawik, Magdalena; Franklin, Kimberly J.; Poulton, Nicole; Lubelczyk, Laura C.; McPhee, Kate; Richardson, Tammi L.; Meerssche, Elise Van; Petri, Brian

doi:10.1007/s10811-018-1541-z

Cited by 6 publications

(3 citation statements)

References 29 publications

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“…Each of the samples was dissolved in distilled water and added to a peptone solution. A serial dilution method as described earlier (19) was applied at 10 folds. The various concentrations were plated onto Potato Dextrose Agar (PDA) media 15 mL and incubated at room temperature (28°C) and observed every 24 hrs.…”

Section: Isolation Of Fungal Organismsmentioning

confidence: 99%

Antifungal Potential of biobased oils from Citrus sinensis Peels and Eucalyptus globulus leaves in vitro against fungal isolates

Abdul-Rahaman

Irtwange²,

Kortse³

et al. 2023

Plant Sci. Today

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Biobased oils found in the leaves of many plants used as coating materials for the preservation of fruits influence the handling of citrus fruits. The effectiveness of these oils on the target organisms is associated with the ability to develop resistant strains. In the present study the antifungal activity of the biobased oils obtained from orange peel and eucalyptus leaves against Aspergillus niger, Aspergillus flavus, Rhizomucor pusillus and Penicillium citrinium was tested at various concentrations in vitro. Sample fruits showing signs of decay during postharvest storage were selected for isolation and identification of the fungi. A serial dilution method was applied at 10 fold for fungi isolates and concentrations were plated onto Potato Dextrose Agar (PDA) media 15 mL. The plates were incubated at room temperature (28 °C) and observed every 24 hrs for possible microbial growth. The pour plate method was used to investigate the antifungal activity of the oils on the test fungi in vitro. The results indicate a continuous decline in the inhibition of the biobased oil treatment during the period of inoculation. The interaction effect of the different biobased oil, their concentrations and the different isolates was significant (P ? 0.05) from day 1 to day 7 during the inoculation. The combined orange peels and eucalyptus leaves at 100% concentration recorded (69.35%) as the highest percent of inhibition which was significantly higher than all other interactions. The least percent inhibition was found with orange peels biobased oil at 1:1 v/v concentration against Aspergillus flavus (24.23 %). Biobased oils at full strength from eucalyptus combined with oil from orange peels demonstrated significant potential against Penicillium citrinium. This research revealed the potential antifungal properties in vitro of the biobased oils against pathogenic fungi. These findings provide the basis for the application of combined biobased oil as an effective antifungal remedy against pathogenic fungi.

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Section: Isolation Of Fungal Organismsmentioning

confidence: 99%

Antifungal Potential of biobased oils from Citrus sinensis Peels and Eucalyptus globulus leaves in vitro against fungal isolates

Abdul-Rahaman

Irtwange²,

Kortse³

et al. 2023

Plant Sci. Today

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“…The concentration of viable cells following alkalization was estimated with the Serial Dilution Culture -Most Probable Number (SDC -MPN) assay (McCrady, 1915;Throndsen, 1978), using the methodology developed and validated by MacIntyre et al (2018MacIntyre et al ( , 2019. The method was modified to use sterile 24-well polystyrene cell culture plates (Sigma Aldrich), based on the approach described by Bernd and Cook (2002), rather than individual tubes.…”

Section: Modified Serial Dilution Culture -Most Probable Number Assaymentioning

confidence: 99%

Assessing the impacts of simulated Ocean Alkalinity Enhancement on viability and growth of near-shore species of phytoplankton

Oberlander,

Burke,

London

et al. 2024

Preprint

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Abstract. Over the past 250 years, atmospheric CO2 concentrations have risen steadily from 277 ppm to 405 ppm, driving global climate change. In response, new technologies are being developed to reduce emissions and to remove carbon from the atmosphere using negative emission technologies (NETs). One proposed NET is Ocean Alkalinity Enhancement (OAE), which would mimic the ocean’s natural weathering processes, raising alkalinity and pH and sequestering carbon dioxide from the atmosphere. The potential impacts of OAE were assessed through an analysis of prior studies investigating the effects of elevated pH on phytoplankton growth rates and by experimental assessment of the pH-dependence of viability and growth rates in two near-shore isolates of phytoplankton. Viability was assessed with a modified Serial Dilution Culture – Most Probable Number assay. Chlorophyll a fluorescence was used to test for changes in photosynthetic competence and apparent growth rates. There were no significant impacts on the viability or growth rates of the diatom Thalassiosira pseudonana and the prymnesiophyte Diacronema lutheri (formerly Pavlova lutheri) with short-term (10-minute) exposure to elevated pH. However, there was a significant decrease in growth rates with long-term (days) exposure to elevated pH. Short-term exposure is anticipated to more closely mirror the natural systems in which OAE will be implemented because of system flushing and dilution. These preliminary findings suggest that there will be little to no impact on a variety of taxonomic groups of phytoplankton when OAE occurs in naturally flushed systems.

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“…The stain cannot distinguish between viable and non-viable cells which still possess enzymatic activity and thus non-viable cells will activate the stain and will be quantified as living, although dying and posing no invasion risk [14]. For evaluation of treatment technologies based on UV irradiation, the Most Probable Number (MPN) method [15] is considered to be a robust viability assessment method [16][17][18][19][20][21]. The MPN method is based on the regrowth potential of organisms in the treated ballast water which is evaluated after an incubation period of two weeks [13].…”

Section: Introductionmentioning

confidence: 99%

Raman spectroscopy as a tool for viability assessment of planktonic organisms in UV treated ballast water

Andreasen

Lundgreen

Holbech

et al. 2020

Vibrational Spectroscopy

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Inter-laboratory validation of the serial dilution culture—most probable number method for enumerating viable phytoplankton

Cited by 6 publications

References 29 publications

Antifungal Potential of biobased oils from Citrus sinensis Peels and Eucalyptus globulus leaves in vitro against fungal isolates

Antifungal Potential of biobased oils from Citrus sinensis Peels and Eucalyptus globulus leaves in vitro against fungal isolates

Assessing the impacts of simulated Ocean Alkalinity Enhancement on viability and growth of near-shore species of phytoplankton

Raman spectroscopy as a tool for viability assessment of planktonic organisms in UV treated ballast water

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