Use of Metabolic Inhibitors to Characterize Ecological Interactions in an Estuarine Microbial Food Web

DeLorenzo, Marie E.; Lewitus, Alan J.; Scott, Geoffrey I.; Ross, Philippe

doi:10.1007/s00248-001-0004-1

Cited by 20 publications

(12 citation statements)

References 43 publications

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“…Vibriosis and photobacteriose are the two most common types of bacterial diseases that can afflict marine, estuarine and sometimes freshwater farming systems and are principally responsible for the greatest impacts already reported in aquaculture (Defoirdt et al, 2007). In addition to the damage posed by the presence of bacterioplankton and mycoplankton in cultures, these microorganisms also interfere in aquatic scientific experiments, generating methodological problems that will be reflected on results (Spencer, 1952;Yetka & Wiebe, 1974), or making it impossible to test hypothesis about the role of bacteria and fungi in an ecosystem or community (DeLorenzo et al, 2001;Trottet et al, 2011).…”

Section: Effects Of Bacteria and Fungi In Culturesmentioning

confidence: 99%

A review on the effects of antimicrobials use in cultures of planktonic organisms: a rocedure for ecological experiments

Agostini

Lopes²,

Macêdo³

et al. 2019

LAJAR

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Many questions about the role of planktonic organisms remain unanswered because of the difficulty in obtaining a medium where bacteria and fungi are not present. Moreover, an excess load of these microorganisms in phytoplankton cultures and zooplankton organisms may cause nutrient competition or diseases and consequently the death of the organisms of interest. For this reason, we reviewed several methods that have been used to obtain axenic planktonic cultures through specific metabolic inhibitors, such as antibiotics and antifungal agents. From 1940 to 2016, most research was related to microalgae and crustacean's cultures, with the antimicrobials: penicillin, streptomycin, chloramphenicol, oxytetracycline, neomycin and nystatin being the most frequently used. The studies that applied antimicrobials agents to planktonic cultures were mainly focused on being able to culture them and to answer questions about the role of bacteria in aquatic communities without previous testing their effectiveness or their effects on non-target organisms. Therefore, this review sought to determine the correct use of antimicrobials in cultures of planktonic organisms to prevent bacterial and fungal growth, without causing damage to non-target organisms and may assist in the implementation of ecologically-oriented scientific experiments where bacterial and/or fungal inhibition is necessary.

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Section: Effects Of Bacteria and Fungi In Culturesmentioning

confidence: 99%

A review on the effects of antimicrobials use in cultures of planktonic organisms: a rocedure for ecological experiments

Agostini

Lopes²,

Macêdo³

et al. 2019

LAJAR

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“…Some authors who found assumptions to be violated nonetheless concluded that eukaryotic inhibitors are useful for revealing qualitative information about microbial food-web dynamics in natural communities (e.g. Sherr et al 1986;DeLorenzo et al 2001). We disagree with this conclusion.…”

Section: Comments and Recommendationsmentioning

confidence: 99%

Testing assumptions of the eukaryotic inhibitor method for investigating interactions between aquatic protozoa and bacteria, applied to marine sediment

Shimeta

Cook

2011

Limnology & Ocean Methods

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Eukaryote-metabolic inhibitors have been applied in published incubation experiments to inhibit protozoans and infer their impacts on bacteria. This method has been used with water-column, sedimentary, biofilm, biofilter, soil, groundwater, wastewater, and sludge samples. The approach rests on assumptions that rarely have been tested: 1) the inhibitor is completely effective against all protozoans; 2) the inhibitor has no effects on other eukaryotes in the system (e.g., microalgae, metazoans) that could influence bacterial activity; and 3) the inhibitor has no effect on bacterial activity. We tested these assumptions for 10 inhibitors used in published studies (amphotericin B, anisomycin, colchicine, cycloheximide, cytochalasin B, fumagillin, griseofulvin, neutral red, nystatin, and thiram) and one combination of inhibitors (colchicine + cycloheximide) using marine sedimentary organisms. Only thiram was completely effective against the natural assemblages of ciliates and flagellates. With some inhibitors, partial inhibition of ciliates was followed by regrowth within 6-24 h or a bloom of a resistant species. Most inhibitors also killed infaunal invertebrates. Five inhibitors were further tested for direct effects on bacteria at concentrations that inhibited ciliates. Each inhibitor altered either esterase activity, denitrification, or both. All inhibitors thus violated one or more assumptions of the method. Conclusions in published work that did not demonstrate the validity of all assumptions underlying this method should not be accepted. Any future attempt to use a eukaryotic inhibitor against protozoans must confirm all assumptions of this method for the particular habitat, species, and processes to which the inhibitor will be applied.

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“…3c). Antibiotics such as penicillin, streptomycin and neomycin have been applied in cultures to inhibit bacterial contamination without negative effects in microalgae (Molina-Cárdenas et al 2016), protozoans (Divan and Schnoes 1982), fish (Forberg et al 2011), mollusks (Howes et al 2014), crustaceans (Agostini et al 2016) and ecological assays with the aquatic community (Middelburg and Nieuwenhuize 2000a, b;DeLorenzo et al 2001;Veuger et al 2004;Cozzi and Cantoni 2006;Trottet et al 2011). The concentration of these antibiotics, however, needs to be evaluated before application in cultures.…”

Section: Resultsmentioning

confidence: 99%

“…This study consisted of experiments to evaluate the effects of different antibiotics (penicillin, streptomycin and neomycin) at different concentrations (Agostini et al 2016;DeLorenzo et al 2001) administered in combination with nystatin (Finley 2012) (Table 1) to plankton cultures. The experimental organisms evaluated were Acartia tonsa Dana 1849 (Copepoda: Calanoida), Amphibalanus improvisus (Darwin 1854) (Crustacea: Cirripedia), and Conticribra weissflogii (Grunow) Stachura-Suchoples & Williams (Ochrophyta: Bacillariophyceae).…”

Section: Methodsmentioning

confidence: 99%

Inhibition of biofilm bacteria and adherent fungi from marine plankton cultures using an antimicrobial combination

2018

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The presence of organic matter in plankton cultures will lead to 10-to 1000-fold increases in bacterial density in less than 24 h. The presence of bacteria and fungi can damage cultivated phytoplankton and zooplankton. These microorganisms can also inhibit experiments investigating the role of these microorganisms in the community and in biological and ecological laboratory studies. The aim of this study was thus to evaluate the effect of penicillin ? streptomycin ? neomycin (antibiotics) in combination with nystatin (antifungal) to select an antimicrobial combination for the inhibition of biofilm bacteria and adherent fungi that is effective and also non-toxic to marine phytoplankton and zooplankton. Acartia tonsa was exposed to different antimicrobial treatments and application routes (culture medium, culture food, both) to evaluate the survival and egg and fecal pellet production endpoints. The same treatments were also applied to measure Amphibalanus improvisus survival and settlement and Conticribra weissflogii growth endpoints. We selected the most sensitive experimental organism and exposed it to some novel antimicrobial combinations. To evaluate the inhibition potential, biofilm bacteria and adherent fungi were exposed to the treatments that were safe for the bioindicator species. A tonsa was considered the most sensitive of all tested organisms. The treatment composed of 0.025 g L -1 penicillin G potassium ? 0.08 g L -1 streptomycin sulfate ? 0.04 g L -1neomycin sulfate showed the best results for A. tonsa and C. weissflogii cultures. No differences were observed for A. improvisus between the treatments. A. tonsa survival rates showed no differences from the control at nystatin concentrations up to 0.005 g L -1 in combination with the antibiotics. The biofilm bacterial density decreased up to 94% under this treatment, and fungal growth was prevented. Species of other planktonic groups should also be tested to improve our understanding of the effectiveness of the treatments proposed.

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Use of Metabolic Inhibitors to Characterize Ecological Interactions in an Estuarine Microbial Food Web

Cited by 20 publications

References 43 publications

A review on the effects of antimicrobials use in cultures of planktonic organisms: a rocedure for ecological experiments

A review on the effects of antimicrobials use in cultures of planktonic organisms: a rocedure for ecological experiments

Testing assumptions of the eukaryotic inhibitor method for investigating interactions between aquatic protozoa and bacteria, applied to marine sediment

Inhibition of biofilm bacteria and adherent fungi from marine plankton cultures using an antimicrobial combination

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