spring'2020 bear hunt with questions'.

[charlie g]

Hi all, we had a surprise snowfall first week of march/20, here in finger-lakes,NY/US. I collected a sprig of over-wintered wild peppermint from my home artificial pond/ stream mesocosm. Water bears plodded on every sample I took for observation from this plant sprigs root substrate!

Ahh but never before did I observe fresh-water tardigrades ( water bears) with jiggling , that is non-fixed globbular inclusions , 'stuffed' into all spaces of the host water bears bodies. These globbular inclusions jiggled with each trudging step the wee bears plodded along with..these globular inclusions even freely jiggled about the water bears head area...this suggests to me these globules are not eggs carried by the water bears. The observed water bears did not manifest any ill-effects behaviorally, from being 'stuffed' with these globular inclusions.

Online 'google access' of the 'microbial and symbiosis studies of Tardigrades (water bears) give rich treatment (including images) of protozoan Pyxidium tardigradum epibiont peritrich ciliates discovered in 1964 by Vander Land...epibiont/ external surface organisms attached to water bears. Fungal agents literally prey on their hapless host water bears in short time frames from initial infection/invasion of the host water bear.

There is a fantastic recent and ongoing research on the bacterial tribes which,unique to each species of water bear, comprise a stable and species-specific microbiome community within each water bear species...just as we 'mega fauna' all carry unique tribes of microbes..our now respected and still poorly understood: 'human microbiomes'.

So please all enjoy my bench encounters with water bears hosting non-fixed inclusions..in all available body spaces...including the bears head regions.

[charlie g]

Target organism collection ( the actual hunt!) protocol: have an initial glass slide with a long narrow swath of water sample placed on it. So ample zones which can be slurped up if a target organism ( in this instance water bear) is observed with scan objective: 4X scan objective in this instance.

Once verified that my water bear was indeed collected/transferred to the slide used for keen observations ( a successful bear hunt!)...I ellected to add methylcellulose ( a vicosity tool useful for meiofauna and protists), I added methycellulose (50/50 with sample fluid volume...and dropped a coverslip on my water bear wetmount slide.

[Chris Dee]

Edit: removed, responded before post completed.

[charlie g]

With Plan apo 10X, and Plan apo 60X, these non-fixed globules ( these inclusions all loosely moved as their host water bear plodded about in ursine fashion.

[charlie g]

These globular inclusions in numerouse water bears observed,,,do not seem like bacteria, or fungal endosymbionts/ infective fungia...are these protozoans?

[daruosha]

Charlie g, I cannot answer your last question, but have to say I enjoyed reading your bear hunting story. Thumbs up.

[charlie g]

Thanks for the kind words, daruosha! I hope to see some of your shared forum microscopy when you have the inclination. Actually I have more images of the free within the body volume...water bear inclusions.

I know it is all psychological...but for myself..literally I can not/ will not harm the water bears...to get isolated inclusions. After observation sessions...these bears are returned to their peppermint root habitat by simple water flushes.

More snow here in finger lakes/NY/US this April'16th. charlie guevara

[charlie g]

To be followed..please consider water-bear microscopy...happy spring'20

[BramHuntingNematodes]

I'm sorry Charlie, but you will never catch a bear like that. Those are traps for a mouse!

[KD Arvidsson]

Hi Charlie! I dont understand all of your story but i am working with it But what i can understand it is very interesting so i " I,ll come back " //KD

[rmb]

Dear KD
Those jiggling things are not parasites or symbiotes. Those are the tardigrades' cells. They float about freely in its insides

[KD Arvidsson]

Thank you rmb! Aha! Very advanced! Not for a beginner. I read your comments and learn in background //KD

[charlie g]

I'm sorry Charlie, but you will never catch a bear like that. Those are traps for a mouse!

Hi Bram ( Georgia/US...how is your spring'20 going, Bram?)...ahh those mouse-traps with fluffy window gaskets to attenuate their 'bite'..are for a pair of feral kittens we took indoors at my dear spouses wishes. They climb and destroyed plants ( plants reside by home windows...kittens climb to look out of said windows..kittens topple cherished plants,,,ah my ginger plant which I started from a produce store ginger-root...now long gone...sigh)...only one of these urchin-kittens has toppled water samples in my microscope bench area ( same scenario..my standing cultures by windows..one errant kitten wants to look out that window...muddy waters on my bench...I curse 'kats'...but the mouse-traps...as 'land mines'..have spared my bench water cultures. I now place such 'attenuated mouse traps in cherished large plants about the the home. The destruction feral cats do outdoors ( I am a bird lover, bird feeder...feral cats totally drove away a nocturnal colony of Eastern Flying Squirrels we enjoyed feeding at our home feeders for years...sigh.)...the destruction feral cats do to home environs outdoor populations of birds I and others enjoy...is horrific...but out door placed mouse-traps would hurt the birds.

Thanks, Bram-hunter...I enjoyed your post...I enjoy the implication that you 'hunt nematodes'! charlie guevara, finger lakes/US

[charlie g]

Dear KD
Those jiggling things are not parasites or symbiotes. Those are the tardigrades' cells. They float about freely in its insides

Hello, rmb, thanks for looking at my water bear thread. IMHO...you have no basis for saying: 'free cells jiggle loosely inside of water bears. I suggest you read for perhaps 20-30 minutes of online content on waters bears...they do not have large 'free cells' loose in their interiors. Happy spring'20 rmb. charlie guevara

[charlie g]

Water bears do not have relativly large loose cells..freely jiggling within their bodiy cavities...especially within their hear region.

[daruosha]

Water bears do not have relativly large loose cells..freely jiggling within their bodiy cavities...especially within their hear region.

The best explanation : ) Especially the first picture. LOL.

[rmb]

I think you will find the body cavity cells of euardigrades (the unarmoured variety that you have pictures of) move passively in the body fluids as the animal moves about. They are metabolically very active. There may be about 200 in each animal. They are also known as coelomocytes and are used to store food reserves. See Ian Kinchins book "The biology of Tardigrades"

[charlie g]

Thank you, thank you..."rmb" for on 4/19/20...posting and kindly correcting me to the reality that all the dense loose inclussions within this group of water bears I collected from an early spring'20 sprig of outdoor wild-pepermint plant in an artificial stream mesocosm...well these 'densely packed water bears' are manifesting their own: coelomocytes, or hemocytes...within these bears large internal body cavities...yes, yes, "rmb"...thanks for your patient correction of my exuberant error to think cellular endosymbionts were spending winter within water bears. Thank you, "rmb" for the literature sources which would aide my understanding of what I have looked at...'looking vrs seeing'...a wonderful dance we all can enjoy! When I hear the oncoming thunder of galloping hoves...I should first think: 'horses'...and not:'zebras'.
Not that it matters...I was simply wrong...what now fascinates me..is why over the years...in 'naked water bears'/ eutardigrades...why is it that I shared so many delightful and long observation sessions with these water bears..all not manifesting these dense populations of coelomocytes/ hemocytes?!! Perhaps if I look back on my water bear observations...none were dead of winter, or early spring samples before ice-melt? Again, thanks for your corrections, 'rmb'.

My 2004, 7th ed "Invertebrate Zoology", Ruppert, Fox, (late) Barnes...pg 512, and my 2001,2nd ed,"Ecological and Classification of North American Freshwater Invertebrates", Thorp, Covich...pg 530 illustration ( figure# 4 depicts: "body cavity cells), pg 535 descriptions of water bears having colorless blood within their body cavity...and circulation of the contained: hemocytes/ coelomocytes/ body cavity cells..by fluid churning as these water bear trudge along.

So are my late spring, summer and fall water bears not manifesting these relatively huge internal 'body cavity cells/ coelomocytes/ hemocytes' cell...or do these 'phagocytic and nutrient storage cells shrink in size and become attached at areas of the water bear to be less noticable?

Bravo, "rmb" for bringing me back from speculations not valid...looking vrs seeing, yes, yes. charlie guevara finger lakes/ Us

[charlie g]

Looking vrs seeing, after I benefitted from "rmb" posts...after I ( duhh) went to my 2004 and 2001 texts...I again sampled the peppermint plant sprigs root-system for a bear hunt ( now spring'20/ Mothers Day all!).

At my bench...note the peppermint culture...note a 'long narrow sample' placed on a slide...note once 4X scan objective locates the target specimen on this elongated droplet of sample...with pipet you slurp up that area of specimen waters, transfer the 'slurp' to slide you will view target organism on. Methycellulose added to slide to which a coverslip will be placed on.

Then it is all about : working distance of your 60X objective...and your 100X dry objective...to observe your hunted/capture/ not injured water bear> charlie guevara

[charlie g]

At scan 4X objective of elongated sample droplet...the target specimen ( in this case a water bear)...the target meiofauna specimen shows up...here I use DF at 4X for water bears.

A slurp and transfer to slide utilized for observations of target...your 4X objective will verify a successful 'hunt'/ transfer. Please note in the higher magnification image: the forward three sets of lobopod/ limbs permit truding traction ( those claws really offer traction) in foward direction...ahhh note the last posterior pair of lobopod limbs point 'backwards/ posterior'...the last pair of water bear lobopods are effective in reverse/ backward movement of a water bear.

Oddly it reminds me of our NYC pet Beagle...at huge Manhattan :'Macy's Thanks Giving Day Parade'..our doggie Bell would only charge at huge parade horses with his front limbs...Bell dragged his back limbs to insure the charge to a huge horse was 'half hearted'..Bell was never intending to catch up to a huge horse which could stomp on him.

Before I lapse into microscopy world views.. my intention with this post is to encourage a predictable...and not difficult protocol for gathering target organisms to to a slide you will observe the neighbor creatures with. charlie guevara

[c-krebs]

Charlie,

I have seen a fair number of tardigrades with the inclusions you have shown us. The most amazing example was about the time I started using a microscope (as an adult) about 16 years ago.
Have a look at this one. It really looked like a plastic bag filled with ice cubes. It moved about normally.

http://www.photomacrography1.net/forum/ ... .php?t=774

[RobBerdan]

Hi - I have only found a few water bears so far this year collected from lichen growing on a mountain ash tree in my yard. I have found two species living there and also found the epizoic peritrich ciliates growing on some and show pictures of Pyxidium tardigradum growing on Ramazzottius oberhaeuseri. Also show some pictures and a movie of Milenisium tardagradum (https://www.canadiannaturephotographer. ... verse.html) - a predatory Tardigrade - I added Jaws music to the clip. I have found three different species of Tardigrade in my back yard. They are fascinating to watch under the microscope.

Iposted an article that might be of interest to anyone wanting to learn more about Tardigrades and how to collect them with lots of links to research articles, pictures and some movies: https://www.canadiannaturephotographer. ... bears.html

I have been looking for Tardigrades the past 4 weeks to try some new photomicrography techniques and so far have only found 5 of them s, and finally one egg.
As it gets warmer I am sure they were will be more. Ice melted off the ponds just 2 weeks ago in the Calgary, AB region.

From your pictures I could not make out the globular inclusions clearly - maybe some arrows would be helpful - they could be cells. At high magnification in the head are single cells that are often visible, also you might try viewing the Tardigrades in polarized light to see if these globular inclusions are birefingent. I have done this and most of the birefringent granules I see are found in the gut.

Thanks for sharing your pictures and experience
RB

[charlie g]

Charlie,

I have seen a fair number of tardigrades with the inclusions you have shown us. The most amazing example was about the time I started using a microscope (as an adult) about 16 years ago.
Have a look at this one. It really looked like a plastic bag filled with ice cubes. It moved about normally.

http://www.photomacrography1.net/forum/ ... .php?t=774

Charles Krebs...I have learned so much, I have enjoyed so much of your kindly shared microscopy over more than a decade...thanks for this kind post you now share...oddly...with your crisp image captures of your waterbear....we all can relish a crisp image of fungal parasitation of a hapless water bear!

The fungal tribe: Entomorporales ( name of this group literally means 'insect destroyer')...this group of fungal parasites are ( for now) mostly pathogens of insects...if you garden, from time to time during growing season you may notice a dead fly, bee, aphid, etc....glued to a berry bush leaf...coated with a fungus...that's the cycle of Entomorphorales pathogens. Nematodes , mites, and water bears are also parasitized by this group of fungi...the more nematodes and water bears from terrestrial mosses are washed out of mosses...the more we may tally fungal parasites of these meiofauna.

Right down to the oil storage droplets...your image suggests fungal attack of your water bear. Perhaps fungi: Ballocephala pedicellate ? thanks for your intrest in water bears, and thanks Charles for the kind post you offer. BTW...if you have other images in the session you observed that hapless water bear...please consider the morphology of your ' square shouldered ice cubes'...vrs phagocytic/ oblate roundish tardigrade ...coelomocytes/ body cavity cells/ hemocytes.

Water bears are thought to be mostly: 'eutellic'...set number of cells once the embryo matures/ once they hatch from egg... their increase in size with several moults is said due to increase in cell size...not increase in cell numbers....so why over years of enjoyment of water bears... why did I never see a water bear 'stuffed with loose cells'..until this late winter/ spring'20 group of water bears from roots of wild peppermint?!! It briefly snowed this 5/12/20 evening...thanks for all your seeing...and looking group! charlie guevara, finger lakes/ US

[charlie g]

Hi - I have only found a few water bears so far this year collected from lichen growing on a mountain ash tree in my yard. I have found two species living there and also found the epizoic peritrich ciliates growing on some and show pictures of Pyxidium tardigradum growing on Ramazzottius oberhaeuseri. Also show some pictures and a movie of Milenisium tardagradum (https://www.canadiannaturephotographer. ... verse.html) - a predatory Tardigrade - I added Jaws music to the clip. I have found three different species of Tardigrade in my back yard. They are fascinating to watch under the microscope.

Iposted an article that might be of interest to anyone wanting to learn more about Tardigrades and how to collect them with lots of links to research articles, pictures and some movies: https://www.canadiannaturephotographer. ... bears.html

I have been looking for Tardigrades the past 4 weeks to try some new photomicrography techniques and so far have only found 5 of them s, and finally one egg.
As it gets warmer I am sure they were will be more. Ice melted off the ponds just 2 weeks ago in the Calgary, AB region.

From your pictures I could not make out the globular inclusions clearly - maybe some arrows would be helpful - they could be cells. At high magnification in the head are single cells that are often visible, also you might try viewing the Tardigrades in polarized light to see if these globular inclusions are birefingent. I have done this and most of the birefringent granules I see are found in the gut.

Thanks for sharing your pictures and experience
RB

Thank you rb/ robert for you kind sharing the link to your wonderful water bear observations, literature notes, and water bear image captures. Ahhh, I can see more than just look now regarding our globally ubiquitous water bears...thanks all for posting! So I wonder...with set number of cells ( mostly water bears are understood as being: eutellic)...why is it that for years of delightful water bear encounters ( some days you eat the bear, some days the bear eats you...I have been told by a bartender years ago)...why do I have observations of water bears with nearly clear body cavities...where did the coelomocytes/ body cavity cells/ hemocytes shrink and attach to in the bears..so the seemed lacking a dense 'stuffing of inclusions'?

I simply have no desire to discet/ squash open a water bear...even after 100X dry-objective observations...I flush the wetmount slide back to the wild peppermint culture jar.

[charlie g]

It is so interesting to me how densely 'stuffed' these waterbears are with body cavity cells/coelomocytes/ hemocytes...and where are these cells when I observed the rather transparent and clear of such inclusions water bears.

I hope we all have a good spring'20, thanks for looking at these freshwater 'bears'. charlie guevara

[rmb]

Charlie
Here is a picture from a slide i made of a tardigrade. You can see the apparatus it uses to feed and just about make out the leg muscles

Ross

[charlie g]

Beatiful microtechnique, Ross, thanks for sharing your work. Indeed I see the leg mucle straps, and claw glands in some of the lobopods in your preperation. I have read that each time a waterbear sheds it's cuticle to increase size of it's nearly set number of adult cells, at each moult, that entire feeding apparatus your slide illustrates well is regugetated/ ejected out of the bear, the class are shed, the terminal gut and anus is shed, the the body cuticle is shed lastly...the process to new claws secreted by each lobopods claw gland, to new feeding apparatus and mouth, new hindgut and body cuticle...well it can take 2 to 5 days. If nutrient and environmental conditions are adverse for a moulting water bear...the next bear after the molt is completed may actually be smaller in size than the pre-molt bear. Thanks for your water bear studies, Ross. charlie guevara, fingerlakes/US

[shawn perez]

Not sure which part of the water bear are you mentioning :/