The use of antibiotics to obtain axenic cultures of algae

Jones, Alvin K.; Rhodes, Muriel E.; Evans, SE

doi:10.1080/00071617300650211

Cited by 33 publications

(19 citation statements)

References 11 publications

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“…Therefore, for specific projects requiring axenic cultures, the culture collection service may engage in cleaning small groups of strains, always keeping the original non-axenic strain as the collection isolate. Special care must be taken with mucilaginous algae, because partners are attached to the sugar coat, usually in a symbiotic fashion [18]. The standardization of quality methods that include the information management of the axenicity status of cultures, are being developed to enable interoperability between different culture collections and laboratories using cultured microalgae [6].…”

Section: Discussionmentioning

confidence: 99%

“…Attempts to remove the co-isolates were developed by mechanical methods and also by the use of antibiotics, but the latter was criticized. The reasons were the difficulty of maintaining viable cultures cleaned by antibiotics [11] and the cell damage inflicted by the antibiotic action, together with the observation that sometimes axenicity was not fully achieved [18]. Important progress in understanding the dynamics of nutrient uptake by the microalgae growing in batch culture and chemostat was achieved by M.R.…”

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confidence: 99%

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Is axenicity crucial to cryopreserve microalgae?

Amaral¹,

Pereira²,

Pais³

et al. 2013

Cryobiology

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Large culture collections of microalgae and cyanobacteria such as the Coimbra Collection of Algae (ACOI) hold unialgal cultures consisting of a population of cells/colonies of a certain species. These cultures are usually non-axenic, as other organisms such as bacteria and microfungi are also present in culture due to co-isolation. Attention has been recently given to partner organisms since studies indicate that some bacteria are important for nutrient uptake of the algal cells, acting as simbionts. Despite this benign effect in the actively growing cultures, when cryopreservation is applied for inactive-stage storage, these organisms may recover faster than the algae, thus affecting their recovery and the viability assessments. In this study, a set of mucilaginous ACOI microalgae were selected, cell features known for their relevance in cryopreservation success were recorded and simple two-step cryopreservation tests were applied. Thawed samples were transferred to fresh culture medium for recovery. Viability was assessed and partner organism proliferation (pop) was recorded. Results were analyzed by t-tests. Statistical models allowed us to support the known tendency for small, unicellular algae with no outer structures to be successfully cryopreserved and the negative effect of vacuoles in the cell prior to cryopreservation. On average cryopreservation with MeOH or Me2SO led to the recovery of nearly half the cells. It was found that the cryoprotection step with MeOH is when pop is triggered and that the use of Me2SO can prevent this effect. Progress on understanding the cultured consortia will assist the improvement of cryopreservation and research using microalgal cultures.

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Section: Discussionmentioning

confidence: 99%

mentioning

confidence: 99%

Is axenicity crucial to cryopreserve microalgae?

Amaral¹,

Pereira²,

Pais³

et al. 2013

Cryobiology

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“…Droop (1967), for example, used 0.04 g L -1 neomycin in combination with other antibiotics in diatom cultures. Jones et al (1973) applied only 0.0000001 to 0.000016 g mL -1 mixed with other prokaryotic inhibitors in cyanobacteria cultures, while Green et al (1967) purified seaweed cultures with 0.0002 g L -1 neomycin mixed with other antibiotics. In addition to the lower resistance to T D compared to T A , T A?N and T D?N had lower survival of non-target organisms, probably due to the nystatin concentration used (0.05 g L -1 ).…”

Section: Resultsmentioning

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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“…Antibiotics are the main tool for producing axenic cultures of marine and freshwater diatoms (Jones et al 1973). Initially, we used them according to published protocols (Kobayashi et al 2003;Bruckner and Kroth 2009).…”

Section: Discussionmentioning

confidence: 99%

“…In particular, they include a-, b-, and g-proteobacteria, bacteroidetes, and actinobacteria (Brachvogel et al 2001;Knoll et al 2001;Schafer et al 2002;Makk et al 2003;Grossart et al 2005;Bruckner et al 2008;Zakharova et al 2010). Conventional approaches for obtaining of axenic diatom cultures employ physical and chemical methods for separating bacteria from diatoms, treatment with antibiotics, and isolation of individual diatom cells for monoclonal culturing (Spencer 1952;Brown and Bischoff 1962;Jones et al 1973;Daste et al 1983;Kobayashi et al 2003;Bruckner and Kroth 2009). Difficulties arising in the course of this work are due mainly to the following factors: secretion of mucus by diatoms, which provides a good medium for bacterial growth; low sensitivity to antibiotics in bacteria contaminating the culture; and poor viability of diatoms after bacterial decontamination.…”

mentioning

confidence: 99%

A procedure for establishing an axenic culture of the diatom Synedra acus subsp. radians (Kütz.) Skabibitsch. from Lake Baikal

Shishlyannikov

Zakharova

Volokitina

et al. 2011

Limnology & Ocean Methods

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Production of axenic cultures of diatoms is a highly relevant task, as these widespread organisms are objects of research in different fields of biology, from ecology to whole-genome studies, and promising as producers of various organic molecules, including biologically active substances. However, culturing of diatoms isolated from natural communities is accompanied by the development of associated microorganisms. We propose a procedure for establishing an axenic culture of the freshwater diatom Synedra asus subsp. radians (Kütz.) Skabitsch. from Lake Baikal. Such a culture has been produced and maintained by subculturing in our laboratory. The procedure involves filtration (to remove free bacteria), detergent treatment (to remove microorganisms associated with diatom frustules), selection of effective antibiotics and their concentrations nontoxic for diatoms (by the disk diffusion method), cell treatment with the selected antibiotic (ciprofloxacin), repeated filtration, and monoclonal culturing of diatom cells. Axenity of the culture was verified microscopically (after DAPI staining) and by molecular biological methods.

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The use of antibiotics to obtain axenic cultures of algae

Cited by 33 publications

References 11 publications

Is axenicity crucial to cryopreserve microalgae?

Is axenicity crucial to cryopreserve microalgae?

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

A procedure for establishing an axenic culture of the diatom Synedra acus subsp. radians (Kütz.) Skabibitsch. from Lake Baikal

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