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Algae Culture - Science topic

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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?
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If the microalgae's color changes from green to white or pale, it means the microalgae are dying or have died. You can confirm this by conducting a chlorophyll test under UV light to verify the growth and health of the microalgae.
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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?
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This seems a common issue. Also, in the paper you mention (Kearney et al., 2022) it was stated that cyanobacteria need helper bacteria so bacterial growth on this medium is somewhat desirable. As for the organic carbon content - natural seawater does contain some dissolved organic carbon, especially if it was procured from a coastal region. This can be reduced by "aging" the water (https://www.researchgate.net/post/Why-must-seawater-be-aged-for-use-in-media-prep). Using a high purity agar (Noble agar, electrophoresis agar) can also help minimize the carbon content.
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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.
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@all The inability of Thalassiosira weissflogii microalgae to grow on agar plates with F2+Si medium, despite successful growth in a liquid starter culture indoors, could be due to several factors. Let's explore some possible reasons and potential solutions:
  1. Agar Quality: The quality of agar used in agar plates is crucial. Ensure that you are using high-quality agar and that it's properly prepared. Agar that is not properly dissolved or sterilized can inhibit microalgae growth. Try using commercially available agar specifically designed for microbiological purposes.
  2. Medium Composition: Double-check the composition of your F2+Si medium. Any errors in preparing the medium can hinder growth. Make sure you're following the correct recipe, including the appropriate concentrations of nutrients and trace elements.
  3. Sterilization: Ensure proper sterilization of the agar plates and medium. Autoclave the medium and agar plates at the correct temperature and duration to eliminate any potential contaminants.
  4. Temperature and Lighting: Microalgae are sensitive to temperature and lighting conditions. Confirm that the temperature and lighting in your lab are consistent with the conditions in which the microalgae were successfully grown indoors.
  5. Nutrient Availability: Check if the nutrients in the F2+Si medium are accessible to the microalgae on the agar plates. Agar can sometimes form a barrier that prevents microalgae from accessing nutrients. Consider pouring a thinner layer of agar in the plates to ensure better nutrient diffusion.
  6. pH: Microalgae can be sensitive to pH levels. Ensure that the pH of the agar medium is within the suitable range for Thalassiosira weissflogii. Adjust the pH if necessary.
  7. Inoculation Density: The initial inoculation density can affect growth. Ensure you are transferring an adequate number of microalgae cells from the starter culture to the agar plates. Too few cells may not be sufficient to establish growth.
  8. Contamination: Verify that your agar plates are not contaminated with unwanted microorganisms that may be outcompeting the microalgae. Sterilize all equipment and work in a clean environment.
  9. Adaptation: Sometimes, microorganisms require adaptation to a new growth substrate or environment. Try streaking the microalgae on fresh agar plates periodically to see if they eventually adapt and grow.
  10. Subculture: If the microalgae on the agar plates are not growing, consider subculturing them into fresh liquid medium. Once they are actively growing in liquid culture, you can attempt to transfer them back to agar plates.
  11. Consultation: If the issue persists, consider consulting with colleagues or experts in algal cultivation or microbiology. They may provide specific insights into growing Thalassiosira weissflogii on agar plates.
Remember that microalgae cultivation can be sensitive, and troubleshooting may involve several iterations to pinpoint the exact issue. Careful attention to the factors mentioned above and patience in experimenting with different conditions should help you successfully culture Thalassiosira weissflogii on agar plates with F2+Si medium.
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I would like to know your suggestions of a suitable protocol to prepare micro algae samples (small size species) for SEM.
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How to prepare macroalgae (mainly seaweed) for SEM?
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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.
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Hi, I have some experience about microalgae culture, If you use BG11 or BBM medium in both cases probably you obtain a good culture, however I suggest you that use another economic alternatives , best regards from Colombia
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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.
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Hi Tomas, OK thanks. All the best, Jules
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growing of scenedesmus sp. green algae
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Thank you Prof Hanelt Dieter
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I am currently working on cultivation of Chlorella vulgaris. I am just curious to know what happens. Thank you in advance.
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There will be pressure development inside the reactor; eventually, algae may die, sparging would be stopped, or the container would explode due to high pressure in the container.
Thanks
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An alternative livelihood can be generated from marine algae.
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Sargassum is a type of seaweed or brown algae (macro-algae) which generally inhabit in shallow water and coral reefs.
  • This algae can be used to make paper, tissue paper or paper bags, as it usually consists of cellulose and hemi-cellulose which are raw materials of paper (preventing cutting of trees to make paper or prevent use of plastic bags).
  • Can use dry algae biomass to burn as fuel, replace coal with dry algae as it will release less carbon dioxide.
  • Can be used to make cosmetics, makeup, pharmaceutical products, sunscreen, anti-aging cream and for hair strengthening treatments. Sargassum is rich in iodine, bromine, mineral salts and vitamins and also have the ability to absorb fats.
  • Seaweed can be used to treat joint pains and skin diseases (burns), as they have high antimicrobial, antioxidant and anti-fungal properties.
  • Seaweed is also used in food and beverage industry, normally used in cocktail drinks.
  • Farmers can use seaweed as fertilisers. they should collect the algae from the coastal zone which will indeed benefit the ocean as it allows a better survival rate of marine organisms. The farmers should let it dry in the sun for two to three days, wash and store for later use.
  • Sargassum seaweed is a nutritious food rich in carotenoids, cellulose, protein, and aspartic and glutamic acids. Sargassum seaweed contains polysaccharies, which support healthy bloody pressure and blood sugar.
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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!
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You may add some growth inducers to your culture media.
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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
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Better to estimate the biomass production of algae per unit time and per unit area, the calculate the carbon content of the biomass. But still, estimation of the amount of carbon that could be decomposed and released into the atmosphere must be calculated. In short, the humification and decomposition coefficients of algae must be considered. The plant that fixes quickly and released carbon by simple decomposition could not be considered a potential carbon sequester.
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Hi Everyone
Could you please name journals which publish review article related to Algae Biotechnology for free within less duration.
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You can try Trends in Biotechnology (IF: 14.343)
Thanks
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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.
  • What could be the cause/causes of these changes? Is it a fungal contamination or a sign that this algae reacted negatively to the light intensities chosen?
  • What would be the best approach to investigate the cause and to determine it?
Any suggestion would be highly appreciated.
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Decoloration est dur aux rzyons solaire ou n'importe quels paramètres qui peuvent dégrader les couleurs
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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.
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chicken manure
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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!
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What I did in past was, centrifuged my culture in BBM and resuspended the pellet. Then I streak plated the sample and performed selective isolation. I was able to drastically decrease the microbial load using this technique.
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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?
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PBR, I solved problem with ulgtrasounds treatmente. It breakes cilliata bot not the D salina cells
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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.
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PS- please excuse the typing and formatting errors in my replies, I am currently typing from a cell phone and cannot see or edit the area in which I am typing the reply.
Also, since it is difficult to add photo files to the “reply” section, I am including some links to videos we have taken of the microscopic appearance/behavior of the cells/spores retrieved from cultured growth in blocks of agar such as this, as well as what has been observed directly in tissue/fluid samples from some of the affected individuals.
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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.
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If you have access to seawater, I would suggest sterilizing by micro filtration (0,22 microns) and then add the well known F/2 medium. It is a complete nutrient medium used at a dose of 1 ml/l and it works fine with Macroalgae.
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I research fresh water algae (chlorella vulgareis).
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My Chlorella vulgaris culture crashed after it was invaded by protists which I couldn't identify. I think they were there in the starter culture. They exhibited amoeboid movement, as shown here under the microscope.
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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?
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Dear Pirooz,
Actually,
Algae and microalgae are excellent aquatic food sources. They also serve as natural filters of nitrogenous wastes. The two types of algae are microalgae and macroalgae. Microalgae are unicellular organisms where as macroalgae are multicellular organisms with simply differentiated body parts. Thus, the main difference between algae and microalgae is the level of classification.
Classification of algae is shown in file here.
But in question you specifying asked about Iran, for this hope this link will help you.
Thanks hope you will satisfy with answer.
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Fatty acids composition of microalgae differed from species to species.
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Please go through the following PDF attachment.
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How to convert Optical density(absorbance) of Algae into cells per ml?
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The first, you should generate a standard curve (optical density vs cells/ml). Then you can be using the standard curve equation to calculate the cell count based on your OD
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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?
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I have been using 1% of DMSO and 5 minutes of incubation for the Chlamydomonas strain cc503.
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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
Phone: +91 8437525941
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Why you are not apply at abroad ?......
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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?
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In press
📷
CALCULATION OF THE SPECIFIC GROWTH RATE OF MICROALGAE
R.P. Trenkenshu
Abstract. The work focuses on techniques of quantifying the specific growth rate of microalgae in both batch and continuous culture. It is shown, that to prove that the specific growth rate is a constant value, both the ratio of two chemical biomass characteristics and dimensional structure of cell population must be constant. Critical analysis of the correctness of using of a logarithmic formula for estimating the specific growth rate (m) of microalgae in the exponential phase of growth of batch culture: m = (lnB2 – lnB1) / (t2 t1). Where B1 and B2 - density (concentration) of culture at a moment of time t1 and t2, respectively. This formula is widely used by most microalgae researchers without proving exponential growth character. Availability of such proofs makes the applying of the logarithmic formula meaningless. Examples of quantitative description of the experimental data obtained for two types of marine microalgae in the exponential and linear phases of culture growth are given.
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Hi
I'm student aquaculture in MSC And I'm going dissertation topic work on the algae.please guide me
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I don't do the algae research. May be later. Thanks you.
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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?
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I recommend remove all suspended solids from your carbon sources (centrifugation, filtration, .....) before cultivation and then use conventional gravimetric determination of biomass dry matter (centrifugation, washing).
Best regards
Vit
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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?
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Also you can do molecular analyses after the staining and storage
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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.
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Hi,
It's depend on the nature of the sample (If was fixed or not)
I recommend the use of hoechst 33342 or Sybr GreenI.
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Among Osmium tetroxide and glutaraldehyde, which one is more suitable for fixing of unicellular algae for SEM. Please provide me protocol if anyone having?
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This is article about cell preparation for SEM
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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?
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Thank you Willy.
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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?
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Dear Mr. Saini,
For Trentepohlia, you can use Bold's Basal Media, Enriched Seawater Media or Waris media combined with Erdscreiber Media or Soil extract with reasonable or blooming success.
For Printzina, use can use the Woods Hole MBL media or WHM media or Optical Haematococcus Media whether as isolates or with its mycobiont Graphis sp. I do not have a working knowledge on the culture of Printzina hence shall not be able to predict the behavioural peculiarities of the species but with Trentepohlia you can expect the best possible results using any of the specific media.
I hope this helps you,
Regards
Dr. Abhishek Mukherjee
P.S. Some links to help you
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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?
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Hi Adam
You could use the cold water from your household water supply. Just add a T to the cold water pluming, make a spiral to place in the aquarium, connect the spiral to the T with a solenoid valve and a thermostat. This should give you sufficient thermal control for this temp range. Just make sure that there is enough circulation around the spiral.
Good Luck
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Which is this aquatic plant species is
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It seems to be a leaf of Ceratophyllum (possibly C. demersum).
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May I have published experimental data and results for the removal of lignin from seaweed (macroalgae) ?
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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?
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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.
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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?
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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. 
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Phenometrics photobioreactors are widely used for laboratorial experiments under controlled condition:
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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?
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Dear all, thanks for the suggestions and sorry for the late answer. I finally did not succeed to grow Nannochloropsis with glucose.
I think the inhibition might be to the difficulty in transporting glucose inside the cell.
Sergio
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Try using nets with thallus, later, try monoline rope
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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?
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A part of the nitrogen is removed by insufflation, so the crops start a nitrogen starvation phase before than occurs with nitrate. You should check the ammonia concentration in the medium.
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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
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Thank you very much!
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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?
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Thank you for your time and guidance, it is very useful paper. 
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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
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Hi, have you any idea about protocol for doing mass spec on microalgal suspensions, to analyse solutes in my media - basically I want to prep my sample straight after harvest, so it can go onto the column without any issues. And then, I want to compare peaks before and after filtering the microalgae feed and compare the peaks that go down in intensity or disappear, and identify those. What are your thoughts on this?
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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! 
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Hi again Thomas
By way of introduction, I studied the ecology and taxonomy of Hydra many years ago. Among other things,  I was fascinated by the ravaging of hydras by Hydramoeba hydroxena, a giant amoeba; and "revenge" by the cladoceran Anchistropus emarginatus, an agressive obligate parasite on Hydra.
I seem to have found a comprehensive answer to your question:
Please check this article and download the free PDF:
Isolation and Cultivation of Endosymbiotic Algae from Green Hydra and Phylogenetic Analysis of 18S rDNA Sequences
Authors: Kovačević, Goran; Franjević, Damjan; Jelenčić, Biserka; Kalafatić, Mirjana
Source: Folia Biologica, Volume 58, Numbers 1-2, December 2009, pp. 135-143(9)
Publisher: Institute of Systematics and Evolution of Animals, Polish Academy of Sciences
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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?
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I endorse what has been suggested. For more specific information, the ciess or genus is required
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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
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extraction fatty acids from algae using frezer drieid samples I am thinking to use fresh samples instead. 
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We used freeze-drying for work we did years ago (see attachments).  Somewhere someone had worked out the process.
Also, I suspect that a NaCl rinsing step before freeze-drying mitigated compounds that would have interfered with the analyses.
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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?
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thank you all, very much.
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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 
3. Chroodactylon
4. Kyliniella
5. Bangia
6. Balbiniana [(How this epiphyte look like/ how to differentiate it from Audoinella) even though i observed many Batrachospermaceae members i never found this.]
7. Rhododraparnaldia 
8. Ballia
9. Bostrychia
10. Nemalionopsis
11. Thorea
12. Psilosiphon
13. Polysiphonia
kindly give me some insight on these genus so that it will help me during my following field visits.
thanks in advance.
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Dear Dr. Elaya,
I got three of them Batrachospermum, Sirodotia, Audoinella.
I think  the sixth genus  should be  Balbiania instead of Balbiniana.
Balbiania shows  cylindrical cells 3-5 µm and 30-90 µm long with many  chloroplasts per cell (Sheath and Sherwood 2011).
Audoinella  typicallyconsists of small, uniseriate branched filaments that contain many plastids/cell. Apparently can modify pigments sufficiently to appear either red or green.Thallus may grow  up to 15 mm high, and composed of more than  50 cells, reddish; basal part consisting of an irregular prostrate system of densely aggregated filaments.
Best regards
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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.   
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To my opinion RAS is the best system for your aims. You can look for RAS and equipment for invertabrates cultivation by following specialized companies (we had very good experience with them when we needed analogical equipment and consultation):
Akuamaks - Fisheries, Aquaculture
address: Rabat Sokak 22/6 G.O.P. - 06700 ANKARA - TURKEY
phone: +90 312 448 09 71 - 448 10 41 - fax: +90 312 448 07 81
 Erwin Sander Elektroapparatebau GmbH
Am Osterberg 22
31311 Uetze-Eltze
Germany
Telephone: +49 (5173) 971-0
Telefax: + 49 (5173) 971-197; www.neomar.de
Good luck! Bakhtiyor
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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 ? 
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Hi Dr. Natarajan Mohan,
If I ve understood well, you don't want to use seawater in the medium for Nannochloropsis..is it correct? In that case, is it necessary to add sources of N and P, as well as Mg, Fe, Ca and other micronutriens like Co, Cu, Mo, etc. You can check the recipies for the ASP medium (Artificial Seawater Medium). Also it is important to include vitamins like biotin or cyanocobalamin (vitamin B12). For more information about media for marine microalgae you can check the book "Algal Culturing Techniques" which is free on line.
Best regards!
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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
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All these look Cyanophycean forms but picture quality is not really up to the mark for any accurate identifications!!!
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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
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Dear Lorenza Ferro
Pls. find the attached files, Help you to prevent contamination
Robert A. Andersen, 2005. Algal Culturing Techniques
Sergey Fedoroff, Arleen Richardson. 2013. Protocols for Neural Cell Culture Springer Science & Business Media,Pp 278. ISBN1475725868, 9781475725865
Teixeira da Silva JA, Cardoso JC, Dobránszki J, Zeng S (2015a) Dendrobium micropropagation: a review. Plant Cell Rep 34:671–704. doi: 10.1007/s00299-015-1754-4
Niedz RP, Bausher MG (2002) Control of In vitro contamination of explants from greenhouse- and field-grown trees. In Vitro Cell Dev Biol Plant 38:468–471. doi: 10.1079/IVP2002316
Regards
Prof. Houda Kawas
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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.
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Hi :D!
Is it the first time you de-freeze Chlorella preserved at -70°C? In my experience, I had preserved Chlorella and Chlamydomonas strains in liquid nitrogen and I didn't have problems. I used DMSO as cryoprotectant. 
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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
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Dear Maria!
I think they are both Dunaliella species. It is possible that the same species (small cells are young  individuals (zoospores) of bigger cells) or 2 different species, it is difficult question.
Best wishes
Tatiana
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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..
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Dear colleague!
You have both genera in your sample. i.e. Eudorina spp. and Pandorina spp. 
Eudorina spp. - colonies always motile, spherical or slightly elongate, of 16-32-64 cells lying some distance from one another and arranged to form a hollow sphere near the periphery of the homogeneous, hyaline, gelatinous envelope. Cells spherical, with or without a beak at the point of origin of the two cilia.
Pandorina spp. - colonies composed of 8, 16, or sometimes 32 cells, held together at their bases to form a sack globular colony surrounded by mucilage. The cells are ovoid or slightly narrowed at one end to appear keystone- or pear-shaped. Each cell has two flagella with two contractile vacuoles at their base, an eyespot, and a large cup-shaped chloroplast with at least one pyrenoid.
Best wishes
Bohuslav
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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.
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That's good! I used Chlorella and Chlamydomonas inmobilized in alginate discs for my undergraduated thesis. The percentage of alginate and calcium chloride was 4% and 2%, respectively. 
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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?
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Wow that's impressive Baik, thanks for the input. 
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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
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Thinking more about your problem, if you are putting in a lot of glucose and not aerating aggressively the pH might drop due to weak buffering of TAP minimal in the presence of rapid respiration of the glucose. This would generate CO2 that converts to carbonic acid. You might want to try stronger aeration/rapid mixing, and/or lower amounts of glucose. TAP doesn't have a lot of buffer capacity. Fine for Chlamydomonas on acetate where respiration isn't that high, but Chlorella on glucose might be pushing it too far. We published a paper in Plant Physiology way back showing that Chlamy on acetate in the light was primarily utilizing the acetate carbon rather than fixing carbon via CO2, even though photosynthesis rates (by O2 evolution) were high. Chlorella could be doing something similar, so a stronger buffer might also be warranted.
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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.
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Make sure you have a pure culture.  The unicellular could be anything.
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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
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One of my recent Honours students also experienced some bizzarre OD (660-670nm) readings behaviour with a TECAN plate reader that measures in a very simialr way to the one you describe.  We were growing filamentous species that adhered to the bottom of wells and it seems the actual read area in the well is very small, so at low cell density the reading area would read areas where there were no filaments and areas where where there was a filament in its gridded read pattern.
Increasing the number of read points and size of read points helped considerably- as did taking multiple replicate readings (3-5x) from each well to establish the level of read variability from the same well. If your cells dont ahere to the bottom- maybe programming some shaking before reading may help randomise cell distribution and even things out a bit.
Additonally- we also found that the cell concentration/OD curves for each species saturated at relatively low OD (roughly 0.5-0.6), but it varied with each species.  So be aware that your capacity to detect increasing cell concentration using OD will have an upper limit, beyond which you cannot detect change.
Condenstation on inside of the well lid also had a significant and unpredictable effect on the readings- maybe because water absorbs increasingly strongly in the near IR range, but wouldnt think it overly dramatic through a short light path (approx 1cm) in plate well.
Hope this helps.
cheers,  Chris
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How can we trigger process of apoptosis in micro/macroalgae?
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It really depends what you specifically want to do and which species you want to kill. But osmotic stress can be quite handy, using distilled water for marine species, and on the same time using specific chemicals (germanium oxyde for fresh water and marine diatoms). On the other hand some fresh water green algae tend to be quite resistant of some chemical treatments (chlorine) as for e.g.: Chlorella sp.
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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...
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lots of good recipes available--try algal culturing techniques by andersen
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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.
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If it´s not trouble with contaminants, certainly the culture conditions are the responsable to the yellow color of the cultures. The BG-11 medium is specific for cyanobacteria. As you are working with coccoid green algae, tray to chance the medium to WC medium (Guillard & Lorenzen 1972). Good loock
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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!
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May be you have to optimize the time so that the cells don't get disrupt.
Better to start from 15 sec then increase with 15 more sec till 5 min for 2 ml sample 
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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?
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I will try centrifuge at lower speed (around 800 g or less for 2 min). When the cells are full of lipid bodies, they are more fragile.
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Identification please!
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Dear Daniel, this is Spirogyra sp. only, but, you have to provide much more clear image. The key to identify this genus is the Unicellular filament, long, chloroplast exist in spiral shape along with many pyrenoids arranged regular intervals. All the best. You check this websites: www.desmids.nl and www.protist.hosei for more images and links related to Desmids. 
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identification
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