Culture Journal, Species: Parvocalanus crassirostris

Re:Culture Journal, Species: Parvocalanus crassirostris (jazzybio13)

Sun, 11 Aug 2013 15:46:24 GMT

mmm. ok! well, I'm going to start up my Iso soon (maybe this week, if I can get an order to arrive!) and see how well I can keep that going. Then get an order of the parvo in and try my hand at it! ... It's threads like this that are well descriptive that encourage me to try my hand at it! lol... Thanks Jim!

Re:Culture Journal, Species: Parvocalanus crassirostris (JimWelsh)

Sun, 11 Aug 2013 12:29:48 GMT

No co-cultures of anything with my Parvocalanus yet.

Re:Culture Journal, Species: Parvocalanus crassirostris (KathyL)

Sun, 11 Aug 2013 09:18:04 GMT

There are some in my ciliate bucket. I'm trying to remove the rotifers from my primary culture. Haven't checked this week.

Re:Culture Journal, Species: Parvocalanus crassirostris (jazzybio13)

Sun, 11 Aug 2013 01:15:20 GMT

updates? have you (or anyone one for that matter) co cultured this with anything? (intentional or not?)...

Re:Culture Journal, Species: Parvocalanus crassirostris (KathyL)

Wed, 03 Jul 2013 00:54:46 GMT

Of course, it does.

Re:Culture Journal, Species: Parvocalanus crassirostris (JimWelsh)

Wed, 03 Jul 2013 00:50:39 GMT

I most certainly do feed my Parvocalanus (and, FWIW, my Acartia) more frequently, but also more lightly, than my Apocyclops cultures.

Quite frankly, I find Apocyclops almost impossible to kill, short of abject warfare. Parvocalanus, on the other hand, are much more temperamental. On the whole, I have found that Parvocalanus need (1) Very light feeding, (2) Very frequently. I feed my various Parvocalanus cultures, in general, at least once daily, and often twice daily. As I have repeatedly said, I tend to err, if at all, on the side of underfeeding. I have found that very light feedings, once or perhaps twice daily, is what does best. Assuming that there is a decent population of copepods in your culture, a feed rate that would amount to about 75,000 cells of Isochrysis per mL per day is about right. Less food, down to even as low a rate as perhaps 10,000 to 20,000 cells of Iso per mL per day, may suffice. Greater feed rates exceeding perhaps 200,000 or 300,000 cells of Iso per mL per day will cause problems.

I have found it important to learn to visually guage the amount of tinting in my cultures, and if I can detect any discernable tinting remaining in my Parvocalanus cultures when it is feeding time, then I skip this feeding, and wait. I have also found that the cultures where I observe the excess tinting have some other issue going on, and they rarely recover by themselves. Often, they need a large (read: 100%) water change (sieve through a 27 micron sieve, and backwash into freshly made, unused, aged saltwater, all the while avoiding any unnecessary exposure to air), and then very light feeding afterwards.

I hope this helps.

Re:Culture Journal, Species: Parvocalanus crassirostris (KathyL)

Wed, 03 Jul 2013 00:38:43 GMT

Jim, do you find that you need to feed the Parvocalanus more and more frequently than the Apocyclops? My Apo cultures thrive on very light, once a week feeding, and my parvo cultures seem to need more...

Re:Culture Journal, Species: Parvocalanus crassirostris (KathyL)

Tue, 02 Jul 2013 22:43:31 GMT

And i either have a lot of dead ones, or lots of molts….Perhaps I am not feeding enough….

Re:Culture Journal, Species: Parvocalanus crassirostris (GreshamH)

Tue, 02 Jul 2013 19:23:47 GMT

the copepedites and ciliates are the same size, hows a mesh standpipe going top help?

Re:Culture Journal, Species: Parvocalanus crassirostris (KathyL)

Sun, 30 Jun 2013 20:36:22 GMT

That's a good question, because I haven't seen a picture of it yet, but its a standpipe with many holes throughout its length, covered with 30 micron mesh. It is attached to a bulkhead that enters through the bottom of the container and angles out with a valve . Obviously this is supported on a stand of some type. The idea is that one can open the valve to drain the container taking old water and ciliates out, leaving copepods in. Then close the valve and refill with pasteurized saltwater.

Problem I have with the design is that I catch ciliates in 30 micron mesh all the time.

Re:Culture Journal, Species: Parvocalanus crassirostris (JimWelsh)

Fri, 28 Jun 2013 23:44:52 GMT

Originally Posted by KathyL

Have you incorporated Andy's standpipe design?

Please refresh my memory about what you are referring to.

Re:Culture Journal, Species: Parvocalanus crassirostris (KathyL)

Fri, 28 Jun 2013 23:26:56 GMT

Have you incorporated Andy's standpipe design?

Re:Culture Journal, Species: Parvocalanus crassirostris (JimWelsh)

Mon, 10 Jun 2013 00:06:47 GMT

It has been exactly a month since my last update of this thread.

My various Parvocalanus cultures continue to do well. I did have a crash of the one 2-gallon fishbowl culture, and I also have had a gross, white bacterial bloom happen in the one 2 liter soda bottle culture, although the copepods don't seem to mind very much. I continue to feed them very lightly, at this point only live Isochrysis, since I've lost both my Pavlova (Monochrysis) and also my Tetraselmis cultures.

The two, three-gallon carboy cultures have both been going for a very long time with very few changes -- not even water changes. I did harvest about 1/2 of the copepods from one of those carboys a few days ago to feed a batch of pipefish fry, but I also just returned the culture water sieved through a 27 micron sieve back into to the carboy.

I've now had these copepods in culture for 5 months. The key seems to be feeding a very, very light level of live Isochrysis, plus perhaps Pavlova and/or Tetraselmis if they are available, once or twice daily. Overfeeding is to be strongly discouraged. If you must err, err on the side of underfeeding. Cultures will do very well if split when population density gets too high, but they will also do well enough if just left alone, and if you do not split.

Re:Culture Journal, Species: Parvocalanus crassirostris (JimWelsh)

Thu, 09 May 2013 12:53:42 GMT

No, actually, I've given up on quantifying my algal densities, after a couple of weeks of watching them with a hemocytometer. I have now learned how to "eyeball" it, and my various cultures are doing quite nicely. Again, as I wrote above, it almost seems as though the less I feed them (within reason), the better they do. I feed very lightly twice a day. Sometimes I forget a feeding, and am always worried that I might be starving them, but then, I seem to get a bloom immediately afterwards!

I've now had Parvocalanus in culture for over 4 months, and have 7 different containers, each with a healthy, dense culture of various life stages. I was unable (or unwilling) to sustain the Pavlova (Monochrysis) and Tetraselmis cultures, and so, I've been feeding a monodiet of Isochrysis for at least the last month. My A. tonsa are also doing similarly well, being treated the exact same way.

Re:Culture Journal, Species: Parvocalanus crassirostris (Little ocean)

Thu, 09 May 2013 06:25:35 GMT

Excellent read and some fantastic pics Jim Welsh,

I was interested in your attempts to measure colour against a chart for algae density, Have you considered the Hanna pocket colour of water checker? I haven't cracked mine open yet but would be very interested to see if this would do the job??

Re:Culture Journal, Species: Parvocalanus crassirostris (clayton447)

Wed, 24 Apr 2013 17:00:00 GMT

13 liters in each tank. 26C for temperature. I used Instant Ocean mixed in deionized water and ran a salinity of 32ppt.

Re:Culture Journal, Species: Parvocalanus crassirostris (JimWelsh)

Wed, 24 Apr 2013 16:58:21 GMT

Interesting. What is the volume of water in the tanks?

Re:Culture Journal, Species: Parvocalanus crassirostris (clayton447)

Wed, 24 Apr 2013 16:56:03 GMT

Well, I have some results. Instead of Phyto-Feast Live, I went ahead and fed the Parvos RGcomplete. I chose RGcomplete because of the ammonia removal component and buffering capability.

So, after 7 days on RGcomplete, they are still alive and I found eggs and nauplii, which means that they are definitely eating the algae and getting enough nutrition to support reproduction. I am also adding heterotrophic bacteria to the system. I will be more specific about the bacteria after I try a few different brands.

As expected, the control tanks outperformed the RGcomplete fed tanks. I started all tanks with 2,000 adults.

The 2 control tanks had approximately 50,000 individuals each after 7 days and were fed live Isochrysis and live Tetraselmis.

The 2 RGcomplete tanks had only 5,000 individuals each after 7 days. These tanks were fed 10 drops of RGcomplete per day. On day 7, I did a 100% water change and will see if the next generation will perform better (creating a strain?). I am going to continue the trials with RGcomplete and try to tweak things as the cultures progress.

I will try to supply updates weekly.

Re:Culture Journal, Species: Parvocalanus crassirostris (JimWelsh)

Wed, 24 Apr 2013 13:31:25 GMT

Originally Posted by clayton447

Hey Jim, great work so far and your attention to detail is staggering. Where do you find the time??? BTW, I am gearing up to test the efficacy of Phyto-Feast Live and RGcomplete on the PC pods. I will be running these two groups along side my control group that is being fed live Isochrysis and Tetraselmis. The trial will be run for a week, starting today, and I will let you all know how it goes. As you probably agree, we need for people to be able to culture this animal without live algae so that breeding of difficult species can be more easily achieved! On a side note, I am also working on a simple methodology for culturing Tigriopus californicus and I will be presenting my results at MBI.

Any updates, Chad?

Re:Culture Journal, Species: Parvocalanus crassirostris (Bongo Shrimp)

Wed, 10 Apr 2013 16:51:42 GMT

Originally Posted by JimWelsh

That is a very good question, Heather. I simply don't know the answer. Maybe Team RMI knows.

I do know that live Isochrysis, Pavlova, and Tetraselmis are all readily eaten by Parvocalanus. I'm not sure about Thalassiosira. Regarding Nannochloropsis, there is this whole issue about its tough cell wall being indigestible by many organisms. I don't know for sure, but I suspect that Nannochloropsis may be indigestible by Parvocalanus.

Whether the condition of the "live" cells in the product is sufficient to be an effective feed for Parvocalanus (e.g., the motile cells are no longer motile, having lost their flagella, as stated here: http://reefnutrition.com/phyto_feast_live.php ) I really don't know.

The Phyto Feast Live page on the reefnutrition.com website (go to http://reefnutrition.com/phyto_feast_live.php and click on the "Phytofeast FAQ" tab) links to the "viability report from the CCMP" document ( http://reefnutrition.com/pdf/CCMP.pdf ) which states that, "...Phyto-Feast Live contains live algal cells that grow and reproduce when placed in an algal culture medium." However, the document doesn't state anywhere in it which specific species of algae grew from the sample provided, so we don't know from this document whether it was, for example, only Nannochloropsis, or if it was multiple species of algae, or all of them, that grew. If, for example, only Nannochloropsis grew, then that is not evidence that all of the various species of algae of interest for culturing Parvocalanus are still "alive" and viable. Also, I'm not sure what the significance is, if any, of the comment in the lower-left corner of each page that says, "Confidential Pursuant to Protective Order - Attorneys Eyes Only - Settlement" (and quite frankly, I feel my eyes have been somehow soiled by seeing this, considering how I feel about attorneys). There are other issues with the CCMP document, such as a typographical error in the expression of the light intensity ("µmol M₂^-1 sec^-2" instead of "µmol M^-2 sec^-1"). As stated, the document indicates that the amount of light provided was increasing exponentially by the second, which is unlikely (to say the least). The reefnutrition.com page also cites this CCMP paper as "more details" to support the statement that Phyto Feast will "...stay alive for several months", but the CCMP paper only documents that the cells were still viable "after 12 days in a refrigerator."

Again, perhaps Team RMI can help clarify some of these questions. I'll see if I can get their attention to this thread. It would be a very pleasant surprise indeed if an off-the shelf product that can be stored in the refrigerator turned out to be a feed that can be used to successfully culture Parvocalanus. I really do hope that Phyto Feast Live is able to accomplish this. That would be GREAT NEWS!

I'm glad someone is going to try Phyto Feast as a food source but my concern, going along with what you said Jim and aside from the fact that the species viable for Parvo would need to be those growing, is that if one isn't growing, won't it will cause problems with the Parvo culture like excess build up of nutrients since you are just adding more biological matter that isn't being used up? That and I also wonder how alive the algae in the Phyto Feast actually is by the time it gets to the end consumer. I mean, "alive" can be used in a very relative way. So how much of the algae is actually alive? And I mean percent of the total biomass alive in the bottle, not which species. If too much is dead, won't you just be adding waste to the culture? And yes of course there will always be some waste but I'm wondering how efficient it will be and if this will cause problems over time. I only point this out because there's no way you are adding the same amount of waste when feeding from the bottle as compared to the fresh culture (correct me if I'm wrong, that's just my logic). I hope I don't sound nit picky but I feel like these could be important factors in the potential success of Phyto Feast as a viable food source for Parvo.