Culture Expansion

[Eatfrenchfries]

Life just happens. Recently finished my contractor position in Portland so I have a lot more free time on my hands. If anyone is hiring let me know haha.

Decided to dedicate a bit more time to get culture production up because meeting demand + working full time + school is just a nightmare. 

Realized that I was tapping out some of my own cultures to keep up with local demand after my own obligations so I made a dedicated system for zooplankton to get around that.

Current production is about 65 gallons of zooplankton + 45 gallons of phyto split across three systems. 

Master Collection - dedicated glass flasks + air filters + air supply + controllable LED diodes + handmade F/2 for the continuous masters. Then there's a 500mL vial with filters for each cultures I swish daily to keep alive in case of scorched earth contingency. Running 3.5 years (1.5 with no critical incidents). Harvest every 5-7 days. Room temp is maintained. Minimal TLC

P. Feed Supply (45g)- phytoplankton + zooplankton for feeding. Also used to grow things on a whim as needed since everything here is interchangeable: culture vessels + T5 + air filters + dedicated air pumps + 1/2 batch 1/2 continuous. This is the system I kept tapping which took a while for some of the less prolific pods to get going again. Harvest every 7-10 days.  Temperature runs amok but nonissue because of diverse feeds. Running 7 months. I keep duplicates of all supplies/vessels to be able to do it on the fly and clean later. 

Surplus Supply-  Last but not least is the newest addition which I will be sharing part of. One of my mentors in the area graciously thought of me when they had a spare 30g RO storage. Thank you very much my good sir. With this I'm basically using a portion of my Primary to seed this vessel and keep one dominant species from taking hold each week. The original plan was to have the vessel producing within a month but after successfully harvesting my pod cysts for the first time I've been able to cut that time in half. This will be the only vessel heated in my whole production line. The reason why is because pods will reproduce faster, but if they're reproducing faster than I need to keep up the feed or they will crash. Will not be changing with fresh saltwater often because I am collecting the detritus to feed my purple non sulphuric bacteria for snow. Hence why this system has 16 gallons of phyto dedicated to keeping #s but not enough to bloom. Leftover phyto from all my other systems will be pooled to do greenwater changes. 

Essentially just a 30gallon container I added a low air supply but still circulates though sometimes theres *no air at all hehe*, plenty of phyto + pods + heater. 

Ambient light comes from the phyto system which are 1/2 gallon glass milk jugs with two snug air holes for rigid tubing(air pushes in so anything going in is pushed out). LED grow light in the day spectrum. Harvest every 5-7 days for phyto and still haven't harvested the zooplankton yet but soon.......

Would love to post videos showing the zoo cultures but sadly unable to. Pictures are a bit more difficult to capture due to their size. 

Just wanted to show that big or small it's all the same. Just the amount of sterilization involved changes. Obviously I go the extra mile to keep things pure since some strains to replace cost upwards of a $100 not to mention its more than a splash to start.

Phyto shown in the pictures are tetra, iso, and dunaliella. Pods in system are apocyclops, tigriopus, tisbe, parvocalanus (just for show, they will either dominate if I maintain feedings of Thal/Iso or they'll die out from not enough). 

The other picture with the coke bottles was mixing enough F/2 to last me a couple months. I usually just make bulk without silicates and then I add in water glass mixed with RODI to the culture water. 

Also just something I thought was cool that a few people asked about when they came by were these reef "mugs" my girlfriend wanted for her nano tank. I designed a reef viewer with a hook handle that can hook on the side of a tank. Made a few that sold quick so if there's a demand I'll make some more. Currently in 3inch and 6inch sizes. 

 

 

 

Edited March 9, 2023 by Eatfrenchfries

[obrien.david.j]

I've been thinking about your home made F/2 fert.   Have you ever watched Cody's Labs.   He made F/2, twice, and an outdoor phyto grow system.   Fun videos

 

 

 

 

[Eatfrenchfries]

I am a little familiar with Cody's labs. Running the same premise of algae + pod cultures outside but since I can't track outside contaminants I mainly use them for personal use. Over winter they'll freeze over but come spring the production speeds up on its own. Sunlight only + culture + food source (dead bugs, banana peel, greenwater) Mainly deal with infusoria, synechococcus, nanno, tig pods for outside cultures due to them being able to withstand fluctuations and temperature extremes. Plus they'll usually remain dominant. Tigs will go dormant until temperatures resume. 

(First time running this in Oregon and it's worked when months are warm. Current pods are hibernating. Used to keep this in California year round, just needed to shade it partially)

 

Mainly I make my own F/2 media to have absolute production control. Not something for the faint if heart because one misstep could ruin future batches. Most prefer to buy it dry in bulk and mix it themselves.

Hand making F/2 should only be done if you have an actual need to alter the formula. Doesn't add up to do it yourself when someone else can do it for cheaper and guaranteed. There's too much sterilization involved to keep it pure. Had a buddy of mine start mixing his own and that batch was contaminated during mixing so it crashed all his cultures in short order with some sort of cyano. 

One batch of my F/2 is mixed stronger than most for most phyto application.  Second batch is made with room for silicates. Third batch contains heavy metals + ammonia. Fourth batch contains extra carbon sourcing. Everything is based on the standard F formula. I play with F/2 and F/3 for culturing purposes. 

[John Vinson]

Do you sell any of your F/2 mix as well? 

[Eatfrenchfries]

@John Vinson I would consider in the future but as of right now I am still developing the refinement process not to mention the formula. 

I could possibly give you a sample to try out for a few batches. 

*I do have an autoclave but there's a lot of technical effort involved in this process. I trust myself but even I'm not perfect. 

[Brittany]

Great talking to you today.   Thanks for the information.

[Eatfrenchfries]

I figured I had the microscope dialed in but getting a digital camera just means more tinkering to be done. Promise I'll get better. I used to use a camera mount for my phone but must say a digital camera does help a lot. 

Just some pictures of the pods I'm working with.

I can get the microscope to focus but the distance of the camera to the lens distorts the image. (Gotta love YouTube videos for troubleshooting)

Parvocalanus crassirostris 

Tigriopus californicus 

Tisbe biminiensis 

Apocyclops panamensis 

*want to point out in the image with the parvocalanus copepod that the colored dots stuck to it is isochrysis / Thalassiosira (phyto). Overfeeding phyto too fast can ruin pod molts since it's too dense for the culture water. 

Parvocalanus has been my favorite since it behaves differently than the other pods as a visible adult. They float in the water column and use their antenna to detect motile phytoplankton. Using their short legs they quickly pounce (teleport) on to its prey. 

 

 

[obrien.david.j]

I'll try a few pictures next time I harvest mine.   So far, I'm happy enough with the cell phone mount I have to take pictures.  But we'll see.

[Eatfrenchfries]

I've had a few questions on why my blend varies in color and isn't always your typical rich green blends but closer to brown most of the time. I culture X amount for contracts and Y for myself that sometimes there isn't always an even ratio in the blend. 

Why does it vary in color? I culture different strains of phytoplankton so,  red + green + brown = brown 

Microalgae Strains: 

Isochrysis - a golden brown 3-8 micron size motile microalgae that is high in fatty acids making it an excellent feed source for filter feeders and gut loading zooplankton. 

Tetraselmis - bright green that get up to 10 -14 microns motile microalgae that is rich in amino acids perfect for feeding. 

Thalassiosira - brown beige ranging in 4 - 32 microns. This is a diatom that requires the addition of silicates to thrive. Beneficial in cycling tanks faster and highly variable as feed for marine organisms due to its large size. 

Nannochloropsis - emerald green 2 - 5 microns nonmotile microalgae. Nutritious feed but has a tough membrane for pods. Excellent source of pigments. 

Porphyridium - ruby red 6 - 10 microns nonmotile microalgae. Highly nutritious (growth) and contains phycoerythrin pigment used to make red filter feeders color pop.

Rhodomonas - strawberry lemonade 9 - 36 microns nonmotile algae. Highly nutritious (growth) feed. Good for photosynthetic organisms and gut loading. 

Symbiodinium (zooxanthellae)- brownish dinoflagellates 6 - 13 microns harbored by photosynthetic organisms like corals, clams, and anemones. Potential to restore coral bleaching. Used as a technical feed for cryptic organisms that require photosynthetic exudates. Currently working with two lab grade and two I harvested myself. 

Synechococcus - forest green cyanobacteria 0.1 - 1.5 microns. Bulk abundant feed source that is fast producing. Used to cover initial larval stages across the board and a staple feed. Somewhat motile, it is believed to possibly compete with red cyanobnacteria found in aquaria. 

 

Diversity in cell size helps cover varying stages of zooplankton life while nutritional profile maximizes yields. I'll say covering cell size is more important than providing optimum nutrition when going for reef bugs. They're pretty hardy when it comes to food if the size is right. Nutrition helps when gut loading for feeding and keeping densities up without competition. 

(Pictures included are 7 of the microalgae strains listed above and one of the bulk pod culture after being freshly fed. )

Bulk Pod Grow after being fed

Symbiodinium #2

Thalassiosira

Nannochloropsis

T

Tetraselmis ( I know the post its say nanno but they've been repurposed, different green)

Porphyridium

Isochrysis

Synechococcus (grown partially outdoor with manual stirring)

Edited April 1, 2023 by Eatfrenchfries

[obrien.david.j]

My Nanno and Tetra continue strong growth.  I hit the 1yr mark of growing Phyto in March.

[Eatfrenchfries]

Having all these live cultures that need daily - weekly input makes traveling anywhere for an extended period of time worrisome to say the least. Usually the zooplankton is fed 2-3x daily with live phyto blend and I need to harvest as well to account for the additional culture volume. Not wanting my dog sitter to do a million and one things I streamlined the process by making algae paste. 

Anyone who brews phyto knows at one point you'll have more volume than you know what to do with. Phyto if maintained properly can last for months. There are a a few methods of concentrating phytoplankton.

Method 1: allow the phyto to sit in the fridge for 2 days and allow it to separate. Skim the water out and keep the phyto on the bottom. Simple. Takes longer for larger cultures to settle. 

Method 2: use a centrifuge and spin the phyto between 3500 - 5000rpm. (4200). Small amount only. Dremelfuge is fun but dangerous. 

Method 3: use 1 micron mesh to filter phyto. Requires vacuum or constant manual stirring to keep from clogging. Most labor intensive and waste material cost.

Regardless of which method you'll end up with phytoplankton concentrate almost like a nice creamy spread. If you use multiple phytoplankton strains it'll come out nice and grainy with odd iridescent hues. 

To get the "paste" you'd use agar or egg whites and it'll thicken up and solidify enough to be manipulated. From there the "paste" is put on wax paper and placed in the freezer to draw out any excess water. 

Microalgae paste can be quite strong so there is a bit of caution to be had. I weigh out one gram pieces to make feeding cultures easy. 

Paste is not live since it is stored frozen and thawed per each use. Removing as much water prior to freezing keeps the cells viable for feeding. Benefits of having a long shelf life and being able to concentrate gallons of phyto into milliliters.

Live phyto is always preferable for feeding so instead of making the paste one could just stop at concentrating the phyto and using that solution. Downsides are the phyto is so dense it need to be properly maintained or it'll be less "live" over time. 

 

 

[obrien.david.j]

15 hours ago, Eatfrenchfries said:

Having all these live cultures that need daily - weekly input makes traveling anywhere for an extended period of time worrisome to say the least. Usually the zooplankton is fed 2-3x daily with live phyto blend and I need to harvest as well to account for the additional culture volume. Not wanting my dog sitter to do a million and one things I streamlined the process by making algae paste. 

...

Method 1: allow the phyto to sit in the fridge for 2 days and allow it to separate. Skim the water out and keep the phyto on the bottom. Simple. Takes longer for larger cultures to settle. 

Method 2: use a centrifuge and spin the phyto between 3500 - 5000rpm. (4200). Small amount only. Dremelfuge is fun but dangerous. 

Method 3: use 1 micron mesh to filter phyto. Requires vacuum or constant manual stirring to keep from clogging. Most labor intensive and waste material cost.

Regardless of which method you'll end up with phytoplankton concentrate almost like a nice creamy spread. If you use multiple phytoplankton strains it'll come out nice and grainy with odd iridescent hues. 

...

So obviously you tried all three methods.  But which one would be your future go to method?

[Eatfrenchfries]

@obrien.david.j

1. (Scalpel) Current and most favorable method for concentrating phytoplankton is through a centrifuge. Least amount of die off and loss of optimal cell nutrition. Downside is having a centrifuge to spin large volumes of phytoplankton is $$$. Most I can spin at a time is between 100 - 200mL depending how much I value my safety. Usually condense 1 - 1.5 gallons down to 18oz for bivalves. Problem with storing any live culture in high density will starve if not cared for properly (use sooner the better). Recently started playing with the notion of creating phytoplankton concentrate starters in test tubes to inoculate larger cultures in case of crashes. 

So far I've condensed all of my phytoplankton strains into 50mL test tubes I keep in the fridge long term in case of a crash and I need a healthy start. I've been able to  use these starters and turn them into 20 liters (about 5 gallons) within 15 - 20 days depending on strain. 

2. (Hammer) I enjoy using a  5um sieve and then a 1um sieve to harvest phytoplankton for making paste. Paste isn't live and loses a bit of nutrition. It is insanely dense and is great for feeding critters in bulk but will foul the water if nothing consumes it. Recommended for heavy filter feeders, zooplankton(pods, brine, rots), less mechanical filtration and more natural filtration the better. 

PSA: don't use dense phyto and oyster larvae at the same time 

Edited April 21, 2023 by Eatfrenchfries

[Eatfrenchfries]

Current strategy revolves around a consistent feeding regimen + a well rounded diet. Got the idea from Algagen to use blood bags fixed with a drip to consistently dose phytoplankton when I am home but easy to store in the fridge when I'm away so it does not spoil. Each pod culture runs on average 50 - 500mL of phyto daily depending what the pod density is running at. The bulk pod container takes 1.5-4L a day. 

Used 5 x 1mL samples of each culture taken before harvest over the course of the month to average out density counts

Tigriopus : 3.8 per mL 

Tisbe: 21 per mL

Apocyclops: 7 per mL

Parvocalanus: 25+ per mL

Each 16oz bottle of pod blend has 4oz (~118mL) from each culture. Going by my worst pod density days and the best I average around 5000 - 7000 pods per container. Ideally the adult to naupli ratio in each master culture should seem like a pyrrhic victory to keep the numbers high or you'll hit a birth/death plateau. 

*Edit (4/28)*- despite the large quantity of pods being displayed on paper the actual viable number is lower. Imagine shoving thousands of critters into a confined space. Can't feed them because they'll waste themselves away. Bigger critters eat smaller critters. Refrigeration is a big no no for copepods. Only Tigriopus can handle the temperature shift making them the most likely to take out all the other pods slowing down. Pods are best kept at room temperature and used immediately. Always check the date. Even if pods are kept species only they can still prey on each other. 

*Realized people like eye candy so no longer using 8oz bottles for pods but going back to 16oz containers. Originally I would sieve 4oz from each culture into the 8oz bottle. Now I no longer need to condense water volume saving me an extra step in the process. 

 

Microalgae

(4 Blend ) ----------

 

Nannochloropsis 

One of the smallest microalgae strains available making it easy to digest for all. It is so prolific that it can even uptake nutrients in the tank making it great for dealing with heavy bioloads. 

• 2-5 microns

• Nonmotile

• High in photosynthetic pigments, fatty acids, and protein

 

Tetraselmis 

The high lipid content is important for fueling growth in marine organisms. A staple feed used in zooplankton cultivation and shellfish (clams, scallops, oysters). Capable of enhancing its nutritional profile in nutrient limited aquaria. 

• 10 - 14 microns 

• Motile

• High in lipids, amino acids, vitamins

• Antimicrobial properties suitable for reef infections

 

Isochrysis 

The gold standard in live food cultivation. Its nutritional profile is essential for enriching zooplankton and rearing most larval invertebrates. The compounds it contains greatly enhance photosynthetic reactions.  It can remain in the water column on its own as long as there are nutrients to consume.  

• 3 - 8 microns

• Motile

• High in lipids, acids, proteins

• Rich in pigments including fucoxanthin (essential feed in photosynthetic coloration)

 

Thalassiosira 

A beneficial diatom that can take up trace metals, excess silicates, and ammonia spikes to speed up the cycling process. Having such a large range in size makes it a great food source for filter feeders and copepods. 

• 4 - 32 microns

• Nonmotile

• Variable cell size allows feed to be utilized in multiple stages of larval development

• Consumes silicates in aquaria which can accelerate the cycling process 

(7 Blend) —-------

 

Porphyridium

One of the few red marine microalgaes making it an exclusive source for red pigments used by filter feeders. The fatty nature of this algae can help boost immune response in marine organisms.

•  6 micron

• Nonmotile

• High in fatty acids, lipids, red pigment (phycoerythrin)

 

Rhodomonas 

Very high in fatty acids and protein that when deprived of nitrogen become even more nutritious making it an excellent feed for cultivating copepods, brine shrimp, and rotifers.

• 4 - 7 microns

• Motile

• High in fatty acids, lipids, red pigment (phycoerythrin)

• Thrives in intense blue light which makes it ideal for reef aquaria

 

Symbiodinium (Zooxanthellae) 

The symbiotic cells that feed a coral via photosynthesis in exchange for a safe place to stay. Commonly used to enhance coloration in clams, restore bleaching in corals and anemones.

• 2 - 4 microns

• Symbiotic dinoflagellates that exist in the photosynthetic tissues of clams and cnidarians (corals, anemones,  etc)

• Coral bleaching is when the zooxanthellae separates from the coral, dosing zooxanthellae can help restore lost pigments and reverse this process 

 

Zooplankton

(Copepods)

*Need to nip this before it gets out of hand but pods do not eat detritus. They contribute to it. 

Tigriopus californicus 

Adults are large and visible to the naked eye making it perfect for picky fish. The bright colors make it easy for fish like mandarins and pipefish to spot. Can reproduce in reef aquaria but difficult to maintain

• Benthic 

• 250 - 1500 microns

• Highly resilient to temperature and salinity

• Rich in fatty acids and pigments

 

Tisbe biminiensis 

A fast breeding grazer of film algae. Even though it is so tiny it produces in such numbers that foraging fish such as wrasse and gobies will be well fed at all times. Perfect for eating leftover foods in the tiniest of crevices. Can reproduce in reef aquaria

• Benthic

• 15 - 200 microns

• Highly reproductive

• Can provides staple nutrition regardless of gut loading 

 

Apocyclops panamensis 

This pod is capable of adapting food sources, environments, and even reproduction rates to survive but will not outcompete other pods. Perfect for rounding out diversity and feeding all fish, coral, inverts. Can reproduce in reef aquaria

• Benthic 

• 70 - 700 microns

• High in acids, pigment, and protein

 

Parvocalanus crassirostris

  The small size makes it the ideal feed for larval fish. Reproduction rates on par with the densest of rotifer cultures but much more nutritious without the need for gut loading.  The cultivation of this copepod paved the way for captive bred mandarins, yellow tangs, orchid dottybacks, melanurus wrasse, butterflyfish, chromis, damsels, etc.

• Pelagic

• 50 - 200 microns

• Highly dense feed akin to rotifers but much higher gut loading potential 

 

—-------

 

Amphipods (Grammarus)

Bigger than copepods but smaller than your clean up crew. These tiny scavengers can  bulldoze all sorts of algae, detritus, and even copepods if there is nothing to keep the population in check. A staple diet for sea horses and a favorite snack among fish. (Larger specimens may prove too large for mandarins)

• 6 - 12mm

• Excellent foragers 

• Feed for larger fish

 

—-------

L - Rotifers 

Tiny food fit for larval fish, SPS, and all sorts of filter feeders because of its gut loading potential. Most commonly cultured for being able to double in population within 24 hours but is nutritionally based upon its phytoplankton diet. 

• 50 - 235 microns 

• High reproduction -> high density feed

• Commonly used in fish fry rearing 

 

Edited April 28, 2023 by Eatfrenchfries

[islandVib3s]

Man, I need to get some pods from you. Which pods don't require phytoplankton and can populate the most in a reef tank? I'm thinking copes?

[Eatfrenchfries]

Not all copepods require phytoplankton as they will eat whatever they can in your tank. If you're looking to increase your numbers and establish a population quicker then the addition of live phytoplankton will help. 

Any of the benthic copepod species will be suitable for populating reef aquaria provided your tank is capable of supporting microfauna. (Tigriopus / Tisbe / Apocyclops)

 

 

Edited April 28, 2023 by Eatfrenchfries

[Eatfrenchfries]

Curiosity killed the cat...….but satisfaction brought it back. 

 

Effective Microorganisms 

 

Nitrifying bacteria:

Biological filtration in reef aquaria. Breaking down organic wastes to be further processed by your system. Without them ammonia levels would rise. We attribute them to be helpful in cycling a tank but in a developed system can be dialed down.

Attempts at culturing this have yielded less than appealing results. Isolating pure strains from reef aquaria involve a culture medium tuned to that specific organism not to mention they are slow growers. 

Inoculum instead is based on whatever was able to reproduce bacteria available in nitrifying reef products. 

Ammonia - Water glass - Molasses - trace elements 

….current cultures are a work in progress. Current methods yield enough to dose back to zooplankton cultures maintaining no water changes for a couple months now. Top off with phyto and RODI plus harvesting for maintenance has been a breeze. No more cleaning.

This leads into my next bit about harvesting phyto + zoo detritus to feed bacteria. 

 

 

Purple Non Sulphur Bacteria

Effective in consuming organic detritus making it now available to the rest of the food chain

Much much easier to get my hands on a few different strains of bacteria not to mention cultivate them. The only thing missing was a growth media which was easy enough to make. Just requires proper sterilization procedure. 

"Why don't you just buy it?" 

Bulk buy would cost me roughly $140 - $160 for 2.5 gallons worth. Not to mention I'd still have to culture it. 

Devising a method to culture my own costs me $5 for the 64oz of rubbing alcohol from Costco, $2 bleach, $5 miscellaneous and leftover nutrient dense matter from my other projects. Managing sterilization and pasteurization to keep it pure. Hardest part about playing with simple organisms is how much even the slightest contamination ruins everything. Utilizing a Still Air Box formerly used in spore prints came in very handy. 

Current culture methods yield  4 x .65g each batch. Upping production is possible for the base bacteria cultivation but nutritional feeding requires mass amount of organic matter rich in nutrients tailored to your aquaria. Attempts in replicating marine snow underway. 

This stuff is just awesome. I highly recommend a read online.

Been giving out samples for people to try for right now. Devising my own culture methods is very taxing so don't expect any quantity for retail yet.

If there is good interest I am considering selling very limited PNSB supply of 8oz bottles for $10 at the Farmers Market to help fund these projects.

 

DM if interested in helping me test out my nitrifying bacteria. 

Edited May 12, 2023 by Eatfrenchfries

[Eatfrenchfries]

Bad news first. I'll be ramping down copepod production as I am redoing my entire zooplankton room. No longer doing 4blend and 7blend phytoplankton. Good news is to streamline the process I just made it 7blend phyto and lowered the price. After doing bulk sales I realized I already do a bunch of culture work weekly so why not make it easier on me and better for you.  Also expanding pod production to include three new species to better support our local breeders. The more diversity I try to maintain forces me to increase biosafety measures. I can't risk any cross contamination since procuring certain single cultures is no longer possible. 

Not exactly sure when I will start producing 7 species pod blend for sale. With the way things are looking it'll be a month before I can start sampling it out. 

 

Euterpina Acutifrons - a pelagic species used for its small naupli size to feed a wide range of fish. Particularly fond of suspension feeding close to surfaces enabling it to jump between water column and sand bed.

 

Acartia Tonsa - a pod that enjoys cooler waters but is quite prolific. Found worldwide and in open waters making it a large part of the food chain

Pseudodiaptomus Pelagicus -  covering all the bases in pod size with these orange giants. Nearly double the size of Tigriopus at 2000-2500 mircrons 

 

 

 

 

A few people have been keen on how I produce copepods. Nothing special to it. 

Make sure ambient temperature is suitable for pods. Salinity varies between 1.018 - 1.026. One or two bubbles per second.

Add pods + appropriate phyto diet (meaty food for big pods)

Feeding frequency is more important than quantity. Depending on pod type will influence how they consume food. I feed at least three times a day if the water clears between feeding.

Some pods will happily eat settled phyto. Some will need live, fresh, nonrefrigerated phyto daily to maintain themselves. (Example: Running 7 x 16oz Iso cultures to cover feeding Parvocalanus daily)

*Know how you pod consumes phyto. Does it hunt, ambush, graze, or need to be coddled?* 

Maintenance involves weekly harvest or water change depending on culture density and water quality. Every so often I will reset the culture by harvesting it and sieving all the detritus out. Hate doing this as the detritus has a large number of pods + cysts so I just throw them into any tank that needs feeding. Replace the water with pure phyto if the pod can handle it. 

 

Don't take it with a grain of salt but do your research please. 

The hardest part about zooplankton is having the appropriate live food in abundance because those little buggers are hungry and messy. 

 

 

P.S. Please do not ask for individual cultures of everything. I offer basic green phytoplankton (2) and zooplankton  (4) species in single culture regularly. That should more than cover the average reefer's needs. 

[obrien.david.j]

39 minutes ago, Eatfrenchfries said:

The more diversity I try to maintain forces me to increase biosafety measures. I can't risk any cross contamination since procuring certain single cultures is no longer possible. 

 

A few people have been keen on how I produce copepods. Nothing special to it. 

Make sure ambient temperature is suitable for pods. Salinity varies between 1.018 - 1.026. One or two bubbles per second.

Congrats on the Zoo-Room-Redo.    I've been wondering/worried about cross contamination between my two pod cultures.  I'm currently using one sieve series (250um->120um->53um stack).   Harvest species 1.   Use them, restart, etc.   Then try to clean the sieve stack with as hot of water as I have.  Plus lots of spraying.   Is it Enough?   Or does your biosafety measure call for dedicated equipment, per species.

On your pod culture technique, how important is light?    I've been playing with the idea of moving some under a stand in my fish room - but it means Super Subtle lighting only.   Some, but not much.    thoughts?

 

[Eatfrenchfries]

So I always work phytoplankton first. Change clothes. Zooplankton 2nd. I use dedicated equipment where possible but rely heavily on rubbing alcohol, autoclave, and bleach to clean between species. Disposable materials help cut down on sterilizing. 

Reef Nutrition opened my eyes to intense safety measures. It really sucks to lose a culture and have to replace it somehow. Also the expansion into gram negative bacteria was reason enough to have stricter methods. 

To me it is all about knowing your variables to be able to say why a culture crashed and possibly learn to correct it. Even then it's hard to give a straight answer. 

Crashes and contamination happen despite all of the precautions. We just do our best to prolong the inevitable and make sure to have backups in place. 

 

 

 

Ambient lighting that has a rough day/night cycle is enough for pods. 

Whenever I work hands on with the cultures I do it in the dark and use light sparingly so as to manipulate them better. They shy away from bright light but will choose points that recieve subdued lighting that barely allows some photosynthesis to occur. (Red light makes their eye pop. I think it messes with their orientation if too intense)

Pods aren't generally photophobic if they feed primarily on phytoplankton. In the ocean there is an abundance of algae cells that end up in the aphotic zone due to water cooling. Pods will migrate between the zones depending on where the food is + age. 

 

The only zoo cultures I have that purposely receive Direct light are adult brine, amphipods, and backup Tigriopus. They are under T5s that heat the water enough to maintain greenwater/macroalgae without a bubbler. (Note: brine and Tig thrive in constant greenwater where others do not. Amphipods are photophobic but love macroalgae)

Adult brine under T5s. Gets turned on at night. 

Edited June 22, 2023 by Eatfrenchfries

[obrien.david.j]

13 hours ago, Eatfrenchfries said:

Whenever I work hands on with the cultures I do it in the dark and use light sparingly so as to manipulate them better. They shy away from bright light but will choose points that recieve subdued lighting that barely allows some photosynthesis to occur. (Red light makes their eye pop. I think it messes with their orientation if too intense)

The only zoo cultures I have that purposely receive Direct light are adult brine, amphipods, and backup Tigriopus. They are under T5s that heat the water enough to maintain greenwater/macroalgae without a bubbler. (Note: brine and Tig thrive in constant greenwater where others do not. Amphipods are photophobic but love macroalgae)

Tigriopus are exactly what I was thinking of expanding.  Big, Meaty, Fun.  Sounds like I'll want to augment the light.    

Thanks!

[Eatfrenchfries]

Managed to isolate some copepods from seawater harvested after the June meeting. For zooplankton I chose an area with low to middle tide zones. The rest of the seawater was collected when the tide began to rise. 

Pods are isolated so they can lay eggs in peace. Those that didn't have eggs were kept together. 

 

caught my eye because it was green and very visible to the naked eye. Very fast. Also it is visibly laden with eggs. Multiple individuals obtained

Different body shape. Much larger. Visible egg sacs. 

very similiar to the first but it seemed to be photophobic whereas the others were drawn to it. Possibly because it is further along developing its egg sacs. 

this picture is from a couple days ago. This pod was separated because it didn't have eggs and now it does. 

I do have moving white flecks on the culture walls so I'm hopeful in raising them. 

-------

Experimenting with new live foods for enticing finicky fish. I must say the yield is much higher than copepods. Main issue is having to gutload them enough to make them worth feeding to saltwater. 

-already have a few cultures available

-feeding portions available soon

 

Daphnia sp. (Water Fleas)

- a freshwater live feed raised between 1.015 - 1.020 salinity. Will survive for hours in reef aquaria but not reproduce.

-gutloaded with Nannochloropsis, Tetraselmis, Thalassiosira, PNSB

- 200 to 3000 micron size. Slow moving with jerking motions. Easy prey. 

Moina macrocopa 

-another freshwater live feed raised closer to 1.018 salinity. Will survive long enough to be destroyed by fish. 

-fed Nannochloropsis, Tetraselmis, PNSB.

-350 to 1000micron. 

 

 

 

[Eatfrenchfries]

Getting ready to start a new batch of PNSB this weekend. If you let me know before Sunday I can make larger quantities. 

Rhodospirillum rubrum becomes colorless in dark + aerobic conditions. I like to store my masters in a cooler with liquid culture medium. Seems to work better that way.  

The culture still has a pink hue from aggregate. Originally I kept everything the way it arrived in agar plates but those proved to be a hassle. Liquid media has been easiest to keep it running. 

Slowly getting better at developing darker pigments in the final blended product. 

Someone told me sucrose wasn't utilized by PNSB but I think the molasses are doing a pretty good job of enriching each batch with B12. Plus I think it's more probable that other bacteria aren't as good at processing sucrose as non sulphur. PSB crushes it everytime when I use treated molasses. Untreated seems to be best for non sulhpur.

Started culturing EM (efficient microbes) again to compost my macroalgae.  Mainly lactic acid bacteria but there's some other stuff as well that came with the original batch (cultivator). Suspect some PNSB and PSB made its way in. 

 

*probably going to make a poll later for what culture techniques people are interested in seeing* 

AMA

[Eatfrenchfries]

A friend of mine came to visit recently and decided to share his 'pretty in pink' phytoplankton with me. Glad to say I got it dialed in. Planning to up production and add it to the blend. See what happens.

dunaliella salina is a extremophile. It lives in salt pools where the salinity can be 4x that of seawater. It does not have much of a cell wall to help maintain osmotic pressure in changing saline environments. It produces an excess of beta carotene (orange pigment) to make up the difference and shield the cell from changing light levels. It is believed that the pink hue comes from bacteria/archaea associated with dunaliella. 

Grown in a tumble has shown weak pink coloration. Stagnant water with heat from a light source has proved to be a much better option. Most likely the biofilm that normally develops in stagnant water is being chewed up by bubbles. 

- 5 to 20 micron

- high in beta-carotene

- more salt equals more pink 

- high nutrient uptake gives potential for a high yield feed. 

- used in brine shrimp cultivation

 

[obrien.david.j]

Love your posts.  We're pretty lucky to have someone with your wealth of knowledge locally!   Keep sharing.

So you don't tumble.  But the picture looks like a 2 liter soda bottle.  How's this work?