Algae Culture - Science topic

I am currently working on culturing Chlorella vulgaris. If it was a contamination, the stock algae should have also been contaminated. But it wasn't, instead it has shown significant growth. I prepared BBM for Standard operating procedure. What would be the reasons?

I am attempting to isolate picocyanobacteria from seawater (pre-filtered with a GF/D membrane) using the pour plate technique and filter plating method (Kearney et al., 2022) on Pro99 and SNAX agar media (0.3% agar) supplemented with cycloheximide to suppress the growth of eukaryotic plankton. After a few days of incubation under a 12/12 h light-dark cycle, I saw many bacterial colonies, which were likely to be heterotrophic bacteria (gram-negative, rod-shaped). For the filter plating method, small mucoid colonies were seen on the filter paper. For the pour plate technique, white turbid colonies were found throughout the agar. Despite extending the incubation period to a month, only a few cyanobacteria colonies were obtained on some agar plates.

I also used Pro99 and SNAX broth in hopes of enriching cyanobacteria before isolation, but the media turned white and turbid instead of green or any other color typically associated with cyanobacteria.

Is it common for heterotrophic bacteria to grow on these media?

According to the recipe to make these media, how is this possible since no organic carbon source is included?

I got a liquid starter culture of Thalassiosira Weisflogii microalgae. While the starter culture grows indoors. But it does not grow on agar plates using the F2+si medium. I am seeking reasons for its inability to grow on the petri dishes and solutions to this issue.

I would like to know your suggestions of a suitable protocol to prepare micro algae samples (small size species) for SEM.

I have been engineering E.coli for few years, but this is my first time transforming microalgae.

I am trying to transform C.vulgaris, using agrobacteirum-mediated trasnformation method. I found a research that used Freshwater Culture Medium (FCM) containing Bold’s Basal Medium (BBM) major salts and supplemented with F medium-trace metals and vitamins for agrobacterium-mediated transfromation of C. vulgaris (https://doi.org/10.1007/s11274-011-0991-0), but I only have BG11 broth in my lab.

Would using BG11 broth significantly affect the transformation efficiency or would it have nothing to do with culture medium?

Thank you.

Spirulina is an exciting agri-food technology, but one concern is that some communities may face obstacles to algae cultivation. Raceway ponds may not be viable in areas facing water scarcity, while enclosed production tanks can be expensive, thus limiting accessibility by poor communities.

growing of scenedesmus sp. green algae

I am currently working on cultivation of Chlorella vulgaris. I am just curious to know what happens. Thank you in advance.

An alternative livelihood can be generated from marine algae.

I have been trying to start some cultures of thermophilic cyanobacteria (in the temperature range of 60-68°C), from environmental samples and from stock cultures, both of which have been stored in a walk-in fridge for 3-4 years. The samples and stock cultures are green in the fridge, but when I try to set them up in the incubator (in 75mL of BG11 medium in flasks, at source pH, under continuous illumination), they photobleach within 3 days. At first, I just set the inoculated flasks into the incubator and raised the temperature with the flasks in it. Then, I've tried several methods to keep the transition more gradual: (1) keeping the inoculated flasks in room temperature in the dark for 1-2 hours prior to moving them to the illuminated incubator and raising the temperature of the incubator while they're in it, (2) covering the flasks, putting them in the incubator, and raising the temperature 10 degrees every hour until they're at the goal temperature, (3) keeping the inoculated flasks in room temperature in the dark for 1 hour, then moving them to the incubator while covered so that they remain in the dark, then raising the temperature 10 degrees every hour, and uncovering them when the temperature reaches 50 degrees.

Could anyone offer me any advice on how to successfully get these cultures growing? Thanks in advance!

To know the carbon fixation rate of a microalgae culture, several bibliographic sources use formulas that consider the biomass change between two times and, knowing the carbon fraction of the biomass and with the molar mass of CO2 and C, we can obtain the carbon biofixation rate of a culture.

However, this raises a question. The variation of the algae culture occurs in the growth phase. But if I have an algae culture in a constant maximun biomass for a year (per example), the carbon fixation rate still aplies over that year?

If that volume of culture fixes X gC02 per day, could I estimate with this formula the annual fixation of C02? Or there is another way to stimate the carbon fixation per year of an algal culture of constant biomass

Thanks in advance for your time

Hi Everyone

Could you please name journals which publish review article related to Algae Biotechnology for free within less duration.

In order to study the optimization of moroccan algae biomass, we're monotoring the growth of 4 different green algae in a laboratory setting in liquid medium under different LED light intensities.

As the culture progressed, we noticed severe discoloration and loss of integrity of one of the macroalgae cultures. However, the rest of the algae species were not affected and remained green.

Any suggestion would be highly appreciated.

I am using TAP and BG11 and comparing the two. In BG11 algae are growing slowly, and in TAP medium they are growing fast but dying soon.

Hello,

I was wondering if there were any methods commonly used to reduce bacterial load in an algal culture. These aren't axenic cultures but the microbial load is a little high, since we'd like to start a microplate experiment soon.

I was thinking a gentle centrifugation to pellet the algae, then to remove the supernatant and resuspend in fresh media. Or to use a cell strainer that allows bacteria but not algae to pass (we have a variety of sizes of algae, though, from chlamy down to blue greens), then keep what is on the strainer.

Or is it better just to start with fresh cultures from our strain bank?

Thanks for any help you can give and take care!

I have contamination from Ciliates in my algae culture (Chlorella vulgaris). I was wondering if lowering the pH would have an effect, say below 4?

Hello,

Our project description page details the unusual origins of our research project/group- looking for the identity of a (likely novel) organism that may be causing a form of chronic illness.

We have successfully cultured an organism from blood donated by affected individuals, and failed to culture it from healthy control blood samples. The organism appears to grow preferentially in the deep matrix of the agar, and only the sporulation stage is visible as surface colonies.

Microscopically, the organism forms structures resembling pseudohyphae, fruiting bodies, and birefringent walled cysts. We hypothesize that it may be related to myxomycetes, oomycetes, apicomplexia and/or other Stramenopile organisms. This is based on morphology and also early metagenomics studies (done on tissue samples rather than the cultured colonies). We are pursuing additional sequencing studies as well asTEM, but have run into complications with DNA extraction (low yields) presumably due to the tendency for the spore nuclei to be encasked in a crystal-like capsule within the agar matrix, as well as the tiny size of the spores/zoospores (1-3 microns)

A recurring finding is that once the organism has infiltrated the agar matrix in the Petri dish, it begins to form spheres and pseudohyphae-like structures WITHIN the deep layers of the agar matrix. Grossly, the agar plate will look like there is no growth for three to six weeks (though samples of agar removed by sterile loop show infiltration of thousands of nuclei-which can be induced to form motile zoospores within 20 min of re-hydration) during this time. Contaminant growth is very rare during this time window. Then, as moisture is removed from the agar, sphere-like objects with a fibrous outer ring, and then fern-like patterns begin to become visible in the dessicated agar. This happens only in the samples cultured from affected individuals, and not in the sham or negative control samples.

I am wondering if anyone else has seen these patterns before in the deep layers of agar (and ideally confirmed presence of an organism by removing/staining blocks of the affected agar). If so, what organism was growing in the agar and, did you find any way of extracting good yields of DNA from the dried/crystallized agar?

PS- photos below are of surface of agar plate/Petri dish as viewed 100x-250x through dissecting microscope. Cultured tissues represented include blood, subcutaneous adipose collected by needle biopsy, and water from a hot tub that might have been a source of infection.

Does anyone have a simple and inexpensive protocol for supplementing seawater for seaweed in vitro culture? I am looking to replace the Provasoli medium with something simpler and cheaper (I have access to sea water). Is just for small/medium scale culture. Thank you very much.

I research fresh water algae (chlorella vulgareis).

Due to the lack of broad algae market in Iran and this image's information, do you think the activities in which the field (macroalgae or microalgae) is more necessary?

Fatty acids composition of microalgae differed from species to species.

How to convert Optical density(absorbance) of Algae into cells per ml?

I want to stain nucleus of green algae by DAPI or Hoechst 33342 by using confocal microsopy. if anyone have appropriate protocol please send to me?

I, Dr. Rachan Karmakar, have done my Ph.D. in 2018 in the field of energy and environment. My areas of specialization are as follows-

1. Biofuel production and characterization

2. Analysis of emission from engines

3. Waste water treatment

4. Algae culture and algae-based technologies

I did my masters in Environmental Science. I have 12 original and published papers including three conference papers (more to come out soon), three years of paid eaching experience and six years of research experience.

I am looking for a post-doctorate position in USA and other countries of America, India, UK, Japan, South Korea and other countries with good research facilities.

Please find my CV attached.

If you are interested to supervise me kindly get back to me or give your email id. I shall contact you through email then. My phone number and email id are given below and in the first page of my CV.

I request every one to help me by sharing and recommending this question, if possible, so that this message can reach maximum number of people and an expert, who can offer me a postdoc, receive the same.

Thanking you,

Sincerely,

Dr. Rachan Karmakar

Email: rachan.in.air@gmail.com

Phone: +91 8437525941

Specific growth rates for cyanobacteria were calculated for exponential phase using selected equation. In the literature, most often appears the equation (and its combinations):

GR = ln(Nt-N₀)/∆t,

where:

N₀ and Nt it is chl-a or optical density values at the beginning and the end of the exponential growth phase

∆t - is the period of the exponential phase expressed in days

any else equations?

Hi

I'm student aquaculture in MSC And I'm going dissertation topic work on the algae.please guide me

I am struggling to find a method to assess the suitability of various carbon sources due to the fact that the carbon sources I am dealing with are quite thick and not clear. I have thought of using dyes (e.g. tetrasodium chloride) but it is possible that the colour of the carbon source will affect the colouration of the culture. I have tried using a spectrophotometer but every carbon source that I test has different thickness and colour and makes the OD measurements not comparable. I have tried cell and colony counts but the results arent very accurate..

Any suggestions?

I am isolates Photobionts from lichens and cultured them. I am getting sufficient biomass on agar plate containing 3N BBM media and broth as well. How can i preserve this biomass for future studies?

I want to microscopic studies of cultured unicellular algal cells. I want to know which dye is most widely used for nucleus staining for algae.

Among Osmium tetroxide and glutaraldehyde, which one is more suitable for fixing of unicellular algae for SEM. Please provide me protocol if anyone having?

I am using 3N BBM broth media in a 250 ml flask for unicellular green algae cultivation. Which condition is very good for faster algal growth? How much days will take for good growth?

I want to culture Trentepholia and Printzina. Which media is best for faster growth? How long time will take for getting good amount of biomass?

Any suggestions on making an aquarium chiller or are there any cheap and reliable ones out there that I could buy?

Has anyone made their own for culturing experiments?

Which is this aquatic plant species is

May I have published experimental data and results for the removal of lignin from seaweed (macroalgae) ?

For example, it is reported that Dunaliella tertiolecta has 36–42% of lipid production in dry weight (dw), while strains like Nannochloropsis can be 46(31–68)%dw, Schiochytrium can be up to 50–77%dw?

Total cellular ATP or NADPH measurement techniques are available but is it possible to measure ATP/NADPH specific to chloroplasts without isolating them? Any noninvasive technique will be very useful.

I am searching through literature to find out the concentration or amount of endogenous copper of Chlamydomonas reinhardtii. However, I cannot find any number. Would someone help me on this please?

We are debating between a growth chamber (adding a shaker) or a regular shaker incubator with lights added on. Any thoughts?

If anyone knows of an incubator specific for algae growth with CO2, temperature and light control, please advise. 

Hi everybody

I'm trying to culture Nannochloropsis oceanica CCMP1779 (axenic) under heterotrophic conditions. For the methodology I'm following previous studies where they cultured other Nannochloropsis spp. and trying to do it under the same conditions. Briefly I inoculated some phototrophically (f/2) growing culture into a new medium made of f/2 and 2g/L glucose and incubated in the dark. However nothing has been growing for 2-3 weeks, while the dark/light control (in the same medium) grows very fast. Therefore Nannochloropsis is clearly not growing heterotrophically and the problem does not come from the medium.

Does anyone have any suggestion on how to grow Nannochloropsis heterotrophically? are there substrates more suitable than glucose for heterotrophic cultivation? Is it possible that other Nannochloropsis spp. can grow heterotrophically while N. oceanica can't?

I need to cultivate Chlamydomonas reinhardtii in an ammonium media with Air + CO2 3% bubbling. After growing very well, my cultures suddenly bleach and die. I do not have this problem with nitrate media.

Do you have an explanation for that?

The media i tried are Bold (with NH4) and TMP (with NH4) with Tris buffer (in order to keep a good pH with CO2 injection).

It is not a problem of N-deprivation because i added nitrate and more ammonium and the cultures also died at the same optical density. Moreover, microalgae shouldn't die following a N-deprivation (they bleach and stay alive for days).

It is not a problem of pH because just before the algae death, it is around 6,5.

It seems to be due to a toxicity but which one? Why don't I have this problem if I bubble only air ?

I really need to cultivate my algae on ammonium media because most of Chlamydomonas mutants are Nit- and cannot assimilate nitrate (i think that we shouldn't use nit- strains anymore because in nature, N is available mainly in the form of nitrate so we should grow microalgae on nitrate media if we want to cultivate them under similar conditions as in nature. But it is another matter of debate :-) ).

Any ideas?

I would like to ask if anyone knows a specific instrument for automatic cell count of an alga culture (P. subcapitata), which also gives information about cell size, morphology and viability. I´ve tried the TC20 from Bio-Rad, but the viability count doesn´t seems reliable. Thank you

Understanding algae responses to stress can help to improve our knowledge about algae adaptation strategies which can be used to engineer tolerant strains. Also, it can be used to manipulate culturing condition to yield the higher amount of secondary metabolites like carotenoid. 

In most of the articles that I am reading it is not mentioned why is important understanding adaptation mechanisms. Are there any other applications other than what I stated?

Basically looking to use mass spectrometry to qualitatively identify the major components of my cell culture, and their relative quantities. But apparently it is a bit tricky with algal cell cultures (cells would be spun down), but the presence of media salts, and potential precipitation of dissolved salts might be an issue. 

Also, I am unsure how to identify the components from the peaks, or if there is a software or database that can automatically integrate the peaks and the m/z fragmented peaks to identify the components?

My media used would either be TAP, BBM-3N-V+ or EG:JM.

Many thanks.

Shawkat

I am trying to isolate endosymbiotic algae from green hydra and culture them. Literatures in the past that claims to succeed in doing so cannot ruling out possibility of being contaminated.

Some basic routine is to wash the hydra and centrifuge it, however, there could still be algae attached to the sticky basal.

Would it be a good idea to chop the basal off and rinse the left part? Any techniques I can kill algae outside while not harming endosymbiotic ones?

Thanks! 

i have used the combo medium to make my culture alive but after 2 or 3 days i found contaimination in it?can any body help me to solve this problem?

I am growing Chlamydomonas (wild type) on 24 well plates and wanting to test its growth rate using chlorophyll fluorescence. The literature says testing every 30 mins for 80 hours will provide a high throughput growth curve. I am looking for any other suggestions on a decent time frame

extraction fatty acids from algae using frezer drieid samples I am thinking to use fresh samples instead. 

Do member of the group arose independently from eubacteria?

i want grow spirulina and chlorella in scale of more than 20 m³ for mass production. but I dont can use medium like F/2 or zarrok and ets beacuse they are expensive and also not very good. do you know what medium is used by companies?

Dear researchers,

 I am doing my research on freshwater red algae, and i have collected many strains from India using techniques learned from Prof. B. B. Chaugule and Dr. Uma rani, but still i could find the representative of following Genus 

1. Cyanidium (will it only occure in Hot water spring)

2. Galderia 

kindly give me some insight on these genus so that it will help me during my following field visits.

thanks in advance.

We would like to culture two species of copepods - Acartia tonsa and Labidocera aestiva. Currently, we have no vendor for aquaculture supplies and are open for recommendation. This will also include culture flasks and shelving for algae culture for food source. Any advice will be helpful as we are looking into feasibility and sustainability of this en-devour as well.   

naturally seawater contains lot of nutrients, however I want to formulate my own commercial medium for my own marine microalgae isolate to grow in open pond/photobioreactors (Nannochloropsis sp), so what are the sole chemical sources are required ? such as Nitrogen, Phosphate etc. Is it necessary to add micronutrients in the medium composition? such as Fe, Zn, Cu, Co, Mo and Mn ? 

It's very small micro-organism I was tried to catch photo and determine it

bright under microscope , it has the same size , the size doesn't increase in google I found photo similar to my organism which called Prochlorococcus 

thank you

Hello,

can someone suggest me which antibiotics are best to get rid of Rhizobium bacteria? And also at which concentration? I would like to add the antibiotics on agar plates and remove bacteria from an algal culture.

Thanks in advance.

Lorenza

Best regards,

We preserved Chlorella vulgaris cultures at -70ºC in glycerol a few months ago. Now, the cultures prepared from recuperated cells don´t growth properly; cultures growth is very slow and has a yellowed color.

Any explanation for this situation?

Thanks in advance.

G.G.

Hi people!

The sampleI (FIXED WITH LUGOL) is from  a temporary lagoon (only 9 cm depht) with high salinity

Conductivity: 172022 μS/cm

pH 6,86

dissov oxig: 2,49mg/l

I am asking about the Volvocales species, there are one bigger and brown colour, and a small green one.

Are they Chlamydomonas? Dunaliella salina???   What do you think??

Thanks in advance

Dear Researchers

 I found this organisms on Roadside stagnant waters. which was forming a floating bloom. they are motile, colonial green algae with eyespot. there i could find two sized colonies weather both sized colonies are same genus or different.or the stage of life cycle. i also want to know about their reproduction method (gametes). 

How to differentiate Pandorina and Eudorina?

In each genus what are key characters are there?

does these organisms are used in pollution indicator index?

i guess these are indicator of organic pollution, is it right?

please help me in identifying the species of these..

we are using calcium alginate immobilized microalgae for various purpose. due to the multiplication of cells inside the beads,  the microalgal cells leaking out from the beads, this  leakage of cells from the beads is the serious issue for us, so if anybody has better idea to prevent the leakage of cells from the immobilized beads. please share with me.

I'm growing Scenedesmus in a small reactor (about 3 litres). After 5 days of cultivation, the alga starts to stick on the walls of the reactor and precipitate out to form a bunch of precipitates. How can I avoid this precipitation?

Hi, I am currently growing Chlorella sorokiniana cells and have had a few problems when trying to grow my cells in TAP (no acetate, but using glucose instead). The cells die after day 2, and the pH drops form 7 to about 3ish. I also wonder why the pH increases from 7 to 8.1 in standard TAP acetate medium for chlorella sorokiniana, but it drops from 7 in TAP minimal media (TAP with no acetate, and pH adjusted to 7 using HCl)? What makes the pH increase in the presence of acetate, but as soon as acetate is taken out (or replaced with glucose), the pH drops! The pH for TAP acetate grown cells increase, whether grown mixotrophically or heterotrophically. Best regards, Shawkat Hussain

I have isolated Botryococcus in a river, when I analysed it through LM it has colonies and mucilage threads connecting. But when I tried culturing it became unicellular but there are some visible threads, but it has a thick cup shape mucilage on the outer cell and takes 14 days to culture while optimising its culture conditions. But later on ill do some ecological studies and phylogenetic studies on that river. But my theory is that the botryococus I have isolated is not in river itself  but the source of it at top or upstream. I hope you can help me.

I have recently started culturing different phytoplankton species in 24-well microplates and taking absorbance measurements using the microplate reader FLUOstar OMEGA. Surprisingly, OD data for all the species have been decreasing over the week, even though it was noticeable that the wells were getting greener and greener, which does not make sense. OD data seem fine when I use the 96-well microplate, so I supposed it had something to do with the 24-well new protocol (maybe the scanning feature, which was the only setting that was changed in the protocol). However, I tested an empty microplate under this new protocol, colouring in black some of the wells, and the results made sense, as OD data were higher in those that were coloured. I don’t know what can be causing this incongruity, can anyone come up with a possible reason??

Thank you very much,

Macarena Blanco

How can we trigger process of apoptosis in micro/macroalgae?

Dear researchers,

I would like to culture Arthrospira and Microcystis which i have collected from local stagnant water. I was just like that collected it so i didnt care about the water pH. But now i wish to culture it, so kindly suggest me good medium composition as well what will be the perfect pH for the both..

Thanks in advance..

Good luck for your research...

Hi all, 

I am having trouble with the growth of my c.vulgaris culture. I am growing them in BG11 medium, the same medium my predecessors were using. I have tried to maintain all the steps and parameters that my predecessors have used, however, after sub-culturing, my algae is turning yellow (Stock culture is green, but after day 2 of introducing in new media, it turned yellow).

There are still growth within the culture as the biomass increases. May I know what is causing this?

Is it my stock culture that is having the problem, or other parameter that i should be concerned about.

Please, can you suggest me a method to separate colonies of 2 or 4 cells into single cells? I am referring particularly to Scenedesmus species. I want to have single cells in my sample to be able to estimate the number of cells through a Cell Coulter counter.

I know heating treatment, chemical or physical treatments can be efficient, but I cannot find any protocol specific for algae.

Thanks!

I am trying to replace the TAP medium with N-free TAP medium after the CW15 has been cultured in TAP medium for 3 days with centrifuge at 1500 g for 5 min followed by pipetting sever time and resuspended into N-free TAP medium. However, the cell been crushed seriously after the replacement operation finished (more than half? I haven’s calculate the crush ratio.). I am confused what's happened with it for the previous experiment went well. Can anyone give me some advice?

Identification please!

identification