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EFFECTS OF THE HYDROGEN ION CONCENTRATION AND OXYGEN

CONTENT OF WATER UPON REGENERATION AND

METABOLISM IN TADPOLES

BY

MINNA ERNESTINE JEWELL
M. A. University of Illinois, 1915.

THESIS

Submitted in Partial Fulfillment of the Requirements for the

Degree of
DOCTOR OF PHILOSOPHY
IN ZOOLOGY

IN

THE GRADUATE SCHOOL
OF THE

UNIVERSITY OF ILLINOIS
1918

TABLE OP COv

P--3
I Intro iuotion •••••••• .---.----.1

II App'tratuu an! V-ithoio ------------- 6

III PreeenlHtion if Data -------------- \q

Regeneration in Baaee ------------- \q

Carbon Dioxida Production and Ravens rat ion in Ba»©« - - - ±\
Regeneration in Aciie - ----------- 17

Carbon Dioxide Production §mi Regeneration in Acid* - - - 21
Halation of Si** to Carbon Dioxide Production a;

Regeneration ----- ---33

Regeneration ia Low Oxygen - ----- ---33

IV Sumwry -------- ----- ---33

V Litenstura Cited ----- ----- - - - 35

VI Tablee - - - --37

VII Figuree ------- ----- ---74

VIII Yite - --114

smcr» or the hydrogen io?; cokcehtratioe aei? oxtoe*

COKTEITT OF f ATER UPOK RE0ISERATI01I
AWD METABOLISM IK TADPOLES

IRTRODUCTIOW

The direct influenoo of toapsraturn upon the rate of development
has Ions 8*6n recognised end hoe recent iy receive J considerable sttsr.tion,
espeeiil'y from the economic entonologlats who are working upon the relation of
wenthor conditions to the titan of emergence of insect pest*. Thit other

environmental factors way eiwllnrly sifMi development la elso recognised
but ae yat the liter ture is ate age r.

Probably no single environmental factor is of greater importance
to aquatic mVrniff than the chemical reaction (hydrogen ion concentrHio
of the water. Shelf ord and Powers (1915) have shown that marine fiehee
are extremaly eeneitlve to slight variations in the hydrogen ion concentration
of the water, onj Well* (1915) h*» shewn the sane to be true of freeh water

fishes.

The first study of tne effect of hydrogen ion concentration

upon the rate of developoent is that of Loeb (1898). He compared the

develcpsmnt of eggs of the sea urchin in normal sea water with those in
ssa eater to which 2 c.c. /lO WaOH per 100 c.c. had been added. At first
no difference was noted but after the thirty-t*o to sixty -four cell stages,
it wes evident the eggs in the alkaline solution hni developed no re rapidly.
The embryos in normal aater were still blestulao when those In the alkaline
water sere plutei, and st the ond of forty-eight hours, though all «ere
plutei, the ones in alkaline *»ter were lerger and farther developed. One, two

2.

or three o.o. n/lC HC1 p«r 100 c.c. was found to retard davelopeont.

Working upon th» eggs of fundulus in fresh water, Loeb found
that 4 o.o. n/l0 IsOH per 100 o.c. caused them to hatch factor. Mora th«n 6 c*:.
retarded development. 1C c.c. allowed only a few of the egge to hatch, while
15 c.o. resulted in failure of any to hatch. 2 o.c. of n/l0 HC1 killed all of
the eggs, while 1 o.c. killed most of them. Losb Attributed these results to an
increase of oxidation by bases and a decrease by noids.

Later Loeb (1914) deolded that the acceleration of development of
the sea urchin eggs in water to which HaOH had been added did not appear until
the thirty-two to sixty-four cell state bsaause at first the concentration was
too high, but after 3 few hours the beneficial effects appeared as tha results
of neutralising the adds produced by tne embryos during development. Theos
conclusions were based upon a comparison of the rate of regeneration and growth
of tubularians in a solution of HsCl, KCl, CaCl2 w»d MgC^'2 in trta proportions
in whioh they occur in sea w:iter, and in the saaas solution to which SaOH, SaHCC3
or H&2 HPO4 lad been added. Althou growth was invariably better when a

base had been added, as the effect) of the weaker bases wars better than those
of the HaOH, Loeb concluded that the results were not duo to the tras OH ions
but to tie ability of tha base to neutralise the acids produced by the tubularian.
Loeb and Waetney (1913 a and b) have shown that bases increase the rate of
oxygen consumption in both the unfertilised and fertilised egge of strongylocon-
trotus purpuratus but that in fertilised eggs this increase in oxygen consumption
is accomplished only by concentrations whicn cause a supprssiion of the phenomena
of development.

floors, Roaf and Whitley (1905) worked upon the effeots of alkalies
and acids and of alkaline and acid salts upon growth and cell division in the
fertilised egge of Echinus eeoulentus. The bases used were TIaOH, KOH, Ca(0H)2

3.

and NH4OH. The acids wore HC1, CK3COOH and C02 and the salts Na2C03, NaHCOg,
Ka2HP04 and NaH2P04. Those workers found th^t bases, exoept NH4OH, give
acceleration of development in low concentrations, while higher concentrations
cheek development and finally kill. Acids all inhibit. The primary factors
affeoting the rate of growth appear to be the variations in concentration of
hydrogen and hydroxyl ions. Thus all of the caustic alkalies are of approx-
imately equal power and there is little or no action of the cations. But in
the case of the phosphates, where the hydrogen-hydroxyl ion concentrations are
comparatively low, there seems in addition to be a specific factor. The
extreme limits of variation of hydrogen and hydroxyl ion concentrations within
which growth is possible are shown to be very narrow, .0015 M. caustic alkali
or .001 M acid practically stopping all development. Whitley (1905) showed
the same relationship between hydrogen-hydroxyl ion concentration and development
to hold for the eggs of Pleuronctes platsssa. He gives three reasons why
alkali is less harmful than acid:

1* Alkali added to sea water is immediately thrown out as insoluble
hydrates or oarbonates.

2. Alkali is constantly used up to neutralize CO2 produced by the

animal.

3. Loeb has shown a low quantity of alkali increases the permea-
bility to 03. This may increase the resisterce of the egg.

Finally Grace Medes (1918) has shown that changes in the composition of the sea

water by concentration, dilution or addition of acids, bases or salts in non

concentrations
lethal cause a retardation of development of the eggs of arbacia punctuUta.

Excepting for the experiments of Loeb in 1898 on Fundulus eggs,

nearly all of the work so far done on the effects of bases and acids on

development has been done upon marine animals. Many of the most serious

4.

Iffic i'.iea in such invaetigationa arias aa the result of tmc ohemioel composition

of aea water and may be largely oli»in*»ted by tha ue« of fraah »*t*r form*.
Thua Haas (1916) hsa eho*n that tha addition of hyirotlaa to *** water reaulte
in a proportion te increase in OH ion ouncantr»tion only aftor all of tha Ca
and Kg have bean precipitated out iu th* form of their basic carbonates.

This diaoovary is of twofold Importance for it asmna in the flrat
place that awch of tMl base added to the eea water la ioRediately thrown
out, ar. 1 in tha seoond pl*co, Ihe disturbance of t^e balance between the salts
of la, K, Ca and Mg, which Loeb (1903) ha* ah own la exceedingly hnrrcful to
marina an ism 1*.

In working with acids too, the use of ae« water presents certain
difficulties, for any strong acid added to aea water will imaediritoly form ealta
with tha liberation of COg. Hence no comparison of the effects of th<> various
mineral acid* in ?oaaibls,a8 the acidity esisiting in the eea water la due to
H2CO3. For theme reason*, end alao to aee whether fresh crater animals are
adjusted to a d iff a rant H ion optimum than marine organisms, the author at flrat
wlahed to undertake a atudy of the effects of bases end acids upon development
and growth of some fresh wster fore. ^inoo, however, there were no eggs or
young larvae available at the time of year »h9n this investigation must ba done,
it w»e decided to uao regeneration which in many way* resembles original growth.
This aeamed further deaimble beesrioe M such studies of regeneration h^ve ae
yet been m=»de Mai it is probable thst comparative studies of regeneration and
growth under varyinr environmental conditions wight throw much light upon the
extent of Mm similarity or dissimilarity of the two proceseee. fhtt la true
of tha effects of hydrogen ion concentration is also true of tat affect of oxygen
i» regeneration. As yet no experimental studies are kmmtn to tnva been awde
on regeneration in varying concent rations of oxygen. As rapid mstaboliam
aal rmmil growth re uaually associated, tha meVibollam experimenta were under-
taken to aea whether any correlation coal! ba traced between the r*te of

s.

regeneration of a port and tue metabolism of the animal ae a whole and whether
eubetances stimulating to oxygen metabolism are, ae Ihitley eu --osed, beneficial
to ths organism. In thle work the author hae been concerned only with the
extrinsic factors of regeneration. The intrinsic factors have bison considered
only ae they arose »s necessary corollaries to the work on environmental effects.

Materials

The tadpoles studied were larvae of Rana alimnta (Daw) collected
from a spring-fed marsh near !luncls, Illinois. Thres collecting trips ware made,
dated October 9, 1917, March 5th and 24th, 1918. This supply was supplemented
during t»e middle of the winter by a few tadpoles purchased fro* a local dealer.
All tadpoles were Identified by means of key to larval amphibia, fright (1914).
V' *. stock of tadpoles was kept in the greenhouse in an albarins tank supplied -rith
running wster to a depth of about 6 incft*s. Tha fosi supplied was f llano . "it ous
green alga*. All tadpoles were kept fasting luring experiments.

Measurements of regeneration were Mfcdt by sain* of ■ caliper and
metal C.Q.S. ruler, obtained from Spencer Lense Company. The tail was the only
organ experimented upon. In making avers&ee there w*ss no elimination of
iniividuala, excepting in one or two instances, and tfeprc not? is made of the
fact tat the aberrant individual included in tne table.

APPARATUS A*D MffTTODS

The oxygen free *atar *«• obtained fro* the water boiling Apparatus

in the vivarium at the University of Illinois which has besn described by

Shelford (193 This apparatus consists primarily of a boiler ir. v L*k the

vater is boiled by <*e*ns of bigfe presaura ateam un.il practically all of the

gasss are driven out and from ahioh it passes into a tank covered by a hood

whs re boiling is continued until the water is freed of gases. The flow cf

water into th? boiler is regulated by a float cork. The oxygen free vater is

withdrawn from the hooded tank, passes through cooling pipae in the ruo» below

where it is used. As It cuasea from the pipes it contains no oxygen (Winkler

method) Ml ie strongly alkaline, due to the C0~ h-ving been expelled by long

boiling and fetish of the bicarbonate having been converted into carbonates. A

series of different oonoentr^tiono cf oxygen was obtained l^j syphoning the

water through a series of half -gallon mason jars (fig.l) through alternate ones

cf which air *as bubbled. Ths water from the last of these was siphoned into

a fivs-inch beaker so that the level of water ir ths Jars co ild not fall below

five inches. It was found by titration that the Oi concentration varied greatly

in the different p».rts of a Jar through which s. stream of air wae passing, but

was relatively constant except at the surface in a Jar through which the water

was be in;: siphoned fc».t no air was passing; consequently the jars containing

the exparlsiental anlmale were alternated with those which were being aerated,

with the result that such oxperi •»■.■•• nt j.ar contained a little ora O3 than the

one preceding.

The flos of air through the water was regulated by passing it

larvfegjl capillary tubes drawn to a fins point and the air pressure was kept

relatively constant by means of a mercury manometer made by bending a D at one

7.

and of & 5 ft. glass tube snd attaching It at the and of the air Una. Mercury
ass than poursd in tube until a prsuaire *%e reach ji »t which »ir would

babble through the Jars at the desired rata.

Dally Ojj deteralnotlonb were aade on the water from g »eh jar
by the Pinkie r eat hod. Seaples f '*e determinations were collected by

introducing a Powera*oa.*plln;7; bottle (Power* 1918) between two jars and allowing
the aster to siphon through it fifteen minutes.

Aa hae been aantiojend above, the oxygen-fro<* water from the boiler
ie strongly alkaline. Intra neutral water watt desired, thle was reraadiad by
aesns of the acidulator (upper left hand of fig. l) a-.ich cone lets of a twelve -
liter aepirated bottle pl*oed in a large earthenware sdlk pen, with the u?per
opening corked securely. T'te acid In ia* pit ., HI timot at %ht loval of the
top of t'.e lower opening in IIm bottle, thua giving a constant pressure in the
siphon which is drawn to a- capillary tube aft the diets! end. 1 floir of

the acii is rag 1a 3i*e si tat capillary tube. ?ha acid drops froa

the oiphon i M separwtory funnel, rough anion it p-iseea to tha bottaa

of the ten-liter aixic£ bottle, vhare it la thoroughly mixed with the water
before enter 'riaental Jara; 1 ,'l '* H3SO4 was uae-i. Vat aoiJity of

*ater could bo ro;: by 00 • the atMkbtf of drop* of i»ci* per ainute

•sn-i ragalatiag tin flow of water throngs taa aisiag tottlt aaaoTdlagly*

In some preliain-iry aaaerlaaata, ordinarily distills! water
obtain* 1 froa the Daaartaaat of Chemistry ffas used. This water ie condensed in
a copper oondenser and paaaes through blocked in tin pipes. It was found not
only to be very toxic to tadpoles but to vary In Its effect froa day to lay so
that tadpoles eiight live sever-*! days in water taken froua one bottle and die
in a few hours in water tnkan froa another. fho saae water redictillsd in glass

8.

supported the Ufa of tadpolsa of ftufo lantlyiflosua. Le Conte through metamor-
phosis. These results are in accord with tits finding of Bullot (1914) an*
Powers (1919 b) who attributes the toxic effect of ordinary distilled tator
to tr-ieea of colloidal coppar taken ftp from the condenser. Powers had shewn
that a goldfish nay live fifty-one 1*ys in '*iter redistilled in glass byt
succumbs quickly if ths minutest trace* of colloidal copper be aided to tha natar.

The water for all of the exporia&ente was onionaed in a six-foot
Jana glass eondanaar attache! to tha second tank of the watar-b oiling apparatus
•stationed above. It was than aerated for 24 hours by a atrosa of air pre*

viously paaaad through sulphuric acid and calcium hydroxid rash bottles. This
dlatllled water »** neutral to Rosalie acid and neutral rai. An enalysia

of the water kJr.J.ly made by tnm water analyst of the State ffater Survey was
as follows:

Physical examination

Turbidity 0

Color 5

Odor V

Residue on evaporation

Total eolide 33 to 42 part* p&r million

Alkalinity as calcium carbomte

Methyl orange 0 to 0 parts per million

Chlorides

At sodium chloride 0 to 0 parts per million

Asmonia nitrogen 4 to 7 parts ■pnr million

Albuninoid ammonia 0*060 to 0.102 parta per million

A oonwon shiner (Notropus blennius) lived in t-tia water (changed weekly) from

Octobar IS, 1917 to Pebniary 17, 1918, or a total of one hundred twenty-two

iaya. As, luring this tim;*, the fish deoreaaal notably in ai»t, ita death

9.
might be attributed to starvation rather than to the toxioitr of the water.
Powers (unpublished) has ohown th.it two goldfishes lived In thie water ninety-five
and ninety-nine days, whsreas in the chemistry distilled w*ter, the length of
life was tiros hundred fifty-two and five hundred ninety-seven eiinutee.

All chemicals ueed were either Marks or Baker's analysed oharaicals.
The acids and basee were standardised at a/lOO, except H3PO4, Na2C03, HaHC03,
which were ^100 tnol.

For the determi:./tion of CO2 production,! pt clamp-top fruit Jars
were used. The capacity of one of thesa J are is about 500 c.c. 400 c.e. of
water or the solution under investigation was put in a jar, then a test tube
containing 10 c.c. n/50 Ba (OH) 2 was inserted so thct the top of the test tube
etood above the level of the vrater, ths rubber ring was adjusted, tho tadpole
put in, and the lid olaapsd down. A stronger solution of Ba(0H)2 *■** used

where acid had been added to tne water or nhnre more CO2 was expected. To
correct for the CO2 in the *ater and in the air above, a control Jsr was prepared
at the same tins and in precisely the same manner, sxoept th"t no tadpole was
put in it, and placed beside the experiment. At intervals, usually 24 hours,

the Ba(0H)2 was nixed aith the ^ster by inverting and rotating the Jar. The
tadpole was quickly reaioved, and the excess Ba(0H)2 titrated with n/l00 HgSOb
using Phenolpthaleir as indicator. The difference li tM amount of acid

required to neutralise the experiment and the control was recorded as the amount
of CO2 (ob a/l00 H2CO3) . produced by the tadpole during the givan tins*

The hydrogen ion concentration was determined by the method of
MeClendon (1916). The solution to be tested w^e placed in a test tube 1 cm.
bore and l/lO o.c. of the indicator solution added. It was then compared with
the color chart by looking down into the tube against a white background. The
indicators used were Butter yellow (Dlmatfcyl amido azobenzene)methyl orange,

10.
4 Br-ehenol - I - pthalsln (Bron Phenol blue) Sietiiyl red, Pcranitro phoncl,
Veutmi red, ?. Rr-thyaol • • « r -thalein (Bron tnyrcol blue) Thymol - 3 - pthniaia
. «ol blue), Phenol pth*la In an-1 Thy»olpthalolr..

.-it lone of Methyl Viol irlla 0 0 (Orange IV) *ere also

prepr.rei but none of the eclat i re so acid ea to eoaa iu their rngo,

Or the oth*r hand, IHi baaic •«!«%] tot ee/or.i %ta range of thle color

"t so th* hydrogen ier. HHHtfaltW of •nlj' ttM aKire dilute solutions ooul J
•a iet*raia».J.

PRFSSmTXOH OF DATA
Before iMflM&nf t&t stp-jrlawate oa regen:*r »t.l >n, a fa* taata were

made to determine arjiroxiw.tsly Wi J to uaa. The flret, a

tadpole woe put into I liter of n/l00 solutions (except H3PO4 0.01 aol)0j asen
of the bases riJa to bo taata I l«ngth of life noted so folloxa •

Base

m

HaOH

B*(GH)2

*ti(0Fi)3

KH40H

Length of life of
tadpole la minutes

50

m

W

H

35

II i I

mo9

m

H2SO4

H3PO4

CH3COOH

Length of life ia
minutes

TO

120

135

135

1M

Tea control* were run. One, itetilled enter f row the metal at 111;
3 waiter *«« uaol In making up the solution*, an 4 I3M Hmnf to e.lbainite the
poaelbUlty of oestotie pressure having produced the effect, a solution ccn-
•Uiilli oi II parte 0.01 «ol. KH^PO^ nn 13 pcfrta 0.01 sol. Ha2 HPO4 which ie
the • tact nautralltT buffer solution of Levy, Rowotrae and M-svrlott (1915)
diluted to 0.01 *ol. Ia the forcer the taiealf lived fir.* days; ia the latter
rt freight da 7a.

The second aeriee wea in .001 normal solutions (except R3PO4 whieh
vae .001 moi.) and gate tha following reaulta:

11.

Bass

:<oh

»aOH

CaCOH).,

s*

Ba(0H)2

VH4OH

Length of
life In days

81

5

30

li

1/6

Aeld

H»03

HC1

H2S04

H3PO4

CH3COOH

Length of
lifo in days

I*

2-

5

8

T

As the table* show, the toxicity of SH^OH is altogether out of proportion to
lta etrength as a base. For this reason, is twe specific toxicities of the
various iona was not within the scop* of the investigation, it was decided to
use only the ssetallic bases and mineral adds.

Tables 1 and 2, experiment I, and 3 and 4, experiment II, give
the regeneration of individual tadpoleo la KOH and Ca(OH)3 reepeotively.
The curves of growth are shorn in figures 2, 3, and 4* These experiments were
oarrled out in finger bowls, eaon containing two tadpoles (one of each sise)
in the amount of base indicated in the table Bads up to 200 e.c. with distilled
water. Tho solution* *ere changed dully throughout the experiment. In plott:
the curves, tho per cent regenerated (taxing the acount removed as 100J&) warn
ussd instead of the actual length rsganerated, because as Zeleny (191?) has
shown, •within wide limits tne length regenerated in the tail of an amphibian
larva is directly proportional to the length removed."

The large range of lew ooncer.tr it iona was used to see if there
might be any acceleration to regeneration by low concentrators, sucm as was
found for development of sea urchin eggs by Loeb (1398) and Moore, Roaf, and
thitley (1905). Aa t«»e growth curves show, no suoh acceleration appeared. In
fact, the Edition of bases produces no appreciable effect until 15 o.c. have
been added, giving a •0075 normal base in which concentration a marked retarda-
tion of rago jn appears in both the KOH and Ca(0H)2«

12.

A comparleon of tablss 1 and 2, *>n •! 3 and 4, brings out anothsr

int»r*stln£ point: That the lRrgor tadpoles (tables 1 and 3) ahow a marked

retardation of regeneration In 15 and 20 o.o. of the baeea, while the smaller

tadpoles (tablaa 2 and 4) ore killed by concent rations which produce reach lata

retardation of regeneration. Th*t while the norwnl regeneration of the

smaller tadpolee 1b much faster than thst of the larger ones, the effect of <»ny

srivsn concentration of baae on regeneration le about the same for both slsss,

being, if anything, more marked la the larger individual while the effect upon

the animal ae a whols, aa shown by the length of life, is greater in case of

the smallsr individual. Frorc this we say believe thnt while regeneration is

affsoted by tne ea«e agents as general metabolism, the two vary independently

and that the thrasholl of toxicity (Powers 1918 b) of a substance to the

regenerating tissues may be above or b^iio* th* threshold of toxicity to the

organism as a whols. This subject of the effect of else upon Regeneration

and Metabolism will be more fully discussed l»ter.

Tho next series of oxrjsriaents (Tables 5,5a and 6. Figures 5, f,

7 and 8) was run in order to determine whether the effect of bau»as upon regener-
ation was due to the action of the hydroxyl lone or whether the ketions or
osmotic prsesuro played an important part. 0. 1 raol. solutions of NagCOa

and KaHCOg were used. The nuaber of o.c. of tho solution used is given in
the first colusm. Thin was amis up to two hundred c.c. with distilled water.
The experiments were carried on In finger bowls, each containing four tadpoles,
and the water was changed alternate days*

In order to assure a uniformity of slae of tadpoles in the various
solutions, the eighty tadpolas used were first divided into four lotu, such
lot containing tadpolos of a uniform slae. One taipola from auch of the four
lots was then put into eaoh of the solutions. Thla method for obtaining
uniformity of six* was ueod in all suooeeding experimante where a large number

13.
of animals was involved. The first, seventh, ani fifteenth dishes were dlatillad
water octrois. In figures 6, 7 and 3 tin 0 line, or our** of growth of the
controls «ae plotted froa the average a of these three.

If iirotio preasurs is the important factor in the concentrations
uael, retardation of regeneration should be the sans, or very nearly the same,
in equi-soleoular solutions of Sa^COa and NaHCOa. If the Va-ion Is the

laportsnt factor, thirty, sirtv, ninety and one hundred twenty c.c. of XaHCX>3
should h-.vo epproxlaately tha tacee effects as fifteen, thirty, fourty-five and
sixty c.c. of Va2C03 respectively, while thirty c.c. each of tfa^COs and 83HCO3
should have the enas effect as forty -five o.c. MajjCOa*

Tables 5,5a an>* 6 show that this Is not the case. Table 4 and
Pig. 5 eho* a progressive and rap 13 deere^ea in regeneration in increasing
concentrations of VagCX^. The tadpoles in 60 an J 75 c.c. died ia four and
three days respectively without h*.vin: .niorgona any regeneration. Table 5 show*
no anrkad decrease in regeneration in increasing concentrations of 9aHC03 until
a .006 aol. solution (120 c.c. in 200) Is reached, although the control gives
the best regeneration. The low psresntage of regeneration of the tadpoles in
30 c.c. oust be attributed to chance as thoao in both the higher and lower
concentrations showed better regeneration.

The curves of regeneration shown in figursa 6, 7 and 3 bring out
•ore clearly the difference in effect upon regeneration of fagCOs and WSHO3.
Thus while 15 c.c. Sa^COs ani 45 c.c. HaHCOs (Fig. 5) depress regeneration but
little more than either of the salts alone, 45 c.c. KA2OO3 and 15 c.c* IfaHX^
(Tig, 3) give practically the easo depression as 45 c.c. fagCOa alone, which Is
inooaparably greater than the depression caused by 45 c.c. RoHCOa alone.

Since there Is this great difference in depreesion of regeneration
between equl -molecular eolutlone of th*» carbonate and bicarbonate, and alnce

14.

■or»owr the bsolsity of the bicirbcntt* (p.H. glwan in last two columns tnbls 5a)
ia sufficient to account for the slight iepression in regens rotten caused by thsa
«t compared to the control, it »»y r.nttslj be concluded th«t the ieprsssion in
regeneration caused by basis is das to t'ne hydroxy! ions. fhla conclusion
ssess ths xaom Justifiable in visw of ths fnot that the concentration.* of salts
used ia ths laltl experiments are three to five times ns great as the concen-
tration* of ths hylroxlds used.

Carb on Bloxlls Pro! notion en J gogene ration in Brnoee

As basic me! la .are generally kno»n to increase oxygen aetsbolism,
it was thought desirable to try a few experiments to see whethar the decrease
in regeneration eoul.t in Any way be correlated *t incre-ia-" in taffbon dioxide

production. 7hs methods employed have already been described.

Table 7, experiment 4, gives the CO^ production in KOH of six
sets of four tadpoles eaoh. Table 0 givso the regeneration of ths esse tad-
poleo. Those i~i tie higher concentrations (30, 35 aad 40 c.c. KOH per 400 e.c.)
died without regeneration. Tig. 9 (drawn froa table 7) repreeent* graphically
the Increase in C03 production with increase! concentration of base and gives
a curwe almost identical with the ourvs shown in Fig. 10, which was drawn froa
the data of Loeb and ITastnay (1913 b) for ths 1 nereis* of oxygen consumption in
increasing concentrations of bases.

Fig. 11 gift's the ourwee of growth for the three sets of tadpolee
which survived. It shoro clearly the decrease both in the mte of regeneration
And the total amount regenerated in increasing concentrations.

Fig. 12 (drnwn froa th-* data of tables 7 and 8) compare* the
effect of the varying concentrations of K3QRI upon the rate of regeneration taken
ss time necessary to regenerate 20, 30 tad 35$ of ths »'rount removed. The
dotted line represents the CO^ per gram per day produced by the s-iae tadpoles.

15.

It will bo noted thnt the rata of ro.jene ration curves and the

carbon Jiorlde curve sre not parallel. Thia, however, would not be expected

em the normal carbon dioxide production is a straight line anl the nortavl rate

of regeneration ia a curvs, progressing more siO#ly as regeneration neare

completion.

It le of interact to note that the curve of regeneration for the

earlieet etage shorn (ZOf> of the amount removed) correspond! more nearly to the
curve of carbon dioxida production Vwn ioea the curve for t»v* second a tags
(30jf of the amount removed) and thle in turn sho*e more similarity than the
curve for the third state (33# of the amount removed). This would suggest
that at the vary beginning of tie process of regeneration the retardation of
regeneration correaponde With the acceleration of oxygen matabolism but that
later the two prooeeeea diverge progreesively, due to the additional inhibition
to regeneration afforded by the presence of the regenerated tiasue.

Tablea 9 and 10, experiment five, give the carbon dioxide pro-
duction and regeneration of sr. other aerlea of tadpoles in KOH In this ease
the normal COg production of each set of tadpolea *ae first determine! an* used
as a basis for comparison of the C02 production under experimental con lit ions*
In graphing the CO^ production (Fig. 14) no. 6 was omitted as these tadpoles
had been injurod by accidental mixing of the Ba(OH)a used to absorb the COg
from the air above the water (Description of apparatus) with the water in which
the tadpolea were.

Tablea 11 an! 1° give the results of a similar series of experiments
using Ca(0H)2 a* the base, and here again one set (So. 4) was spoilt* .1 by
accidental contamination of the solution with Ea(OH)£. Figures 13 and IS are
the ourvea of regeneration in KOH and CaOH raspactlvely; figures 14 and 16
shoa the rate of regeneration so time necessary t-> regenerate a given per cent
of the awount reeoved, and the C0„ production. While these curves are not

16.

comparable, tha regeneration curves being plotted ??ith ordinate as concent mtlon
and asscissa as time, an i tne COg cunreo being plotted with orlin its ae con-
centration *m ■' *o«ciBea as cc. COo as 0*01 WBSCO3, still they do serve to shoe
that as CO2 production is increased, regeneration is decreased progressively.

Child (1911) has shown th-*t for pl^.naria the rate of regeneration
le proportional to tlM nta of met-sbolisw: and thit consequently younger or
entailer worms regenerate store rapiily than the larger on^s. In tadpoles, as
has already been pointed out, the rate of regeneration is greater in the soallsr
indiviiuals. The fact, then, that a decrease in regeneration is correlated
with a rles in carbon dioxide production in basic media must be regarded as
shoving tnat the increased oxygen aetabelism due to bases is a destructive
metabolism and not indicative Of any stimulation of tha normal functions of
the organ ism.

It will be notad that in both of the above experiments (Fig. 13
and 15) the regeneration of tha tadpoles in watar containing ten cubic centimeters
of the base nvade up to 400 was better than that of the controls. It will also
bs seen by examination of the data (tables 9 and 10) that while the initial
hydrogen ion concentration of these solutions is decidedly on the basic side
of neutrality (P.H. aoout 9), that by the time the solution was changed, it was
slightly acid (P.H. about 6.3). For this reason another series of experiments
was planned, using- trv» saaa concentrations, but a larger volume in proportion
to the size of the tadpolas and changing the solution oftener. two parallel
eeries were run, each containing five small tadpolae in one liter of distilled
water, ,90025 H. EaOH ani .0005 B. HaOH(25 0.0. and 50 o.c. 0.01 ». par liter).
The experiments were curried out in two-quart mason jars kept tightly sealed
ani containing test tubes of Ba(0H)2 solution to absorb the CO^ from the air
above the water. The results of this experiment are given in t-ible 13. Due

17.
to the CO, produced by the tadpoles the distilled water became acid and the
NaOH solution less basic. However, it was no longer neutralized. The retarda-
tion to regeneration (Fig. 17) is very marked in 25 o.c. NaOH per liter while
50 o.c. per liter proved fatal in two d.^ye.

These results are in accord with the suggestion of Loeb (1904)
that the beneficial effect of the addition of a base is due, not to the presence
of hydrozyl ions, but to the neutralization of acids produced by the animal, and
show that a P H between 3 and 9 is markedly detrimental.

Regeneration Ir Acids

As the previous experiments show, neutrality is more favorable
for regeneration in talpoles than bacisity. Is the same thing true of acidity?
A complete series from acids, almost certain to H«rw detrimental effects to bases

•n to be harmful, was formed by using 25, 20, 15, 10 and 5 c.c. of 0.01 mol.
H3P04 and 5, 10, 12.5, 15, 17.5 and 20 c.C. of 0.01 H NaOH made up to 200 c.e.
with 5 o.c. 0.01 mol. neutral Na2HPO| ^ KH2P04 mixture as a neutral control as
well as the usual distilled water controls. The solutions were changed daily.
It was hoped in this ray to locate approximately the optimum hydrogen ion con-
centration for regeneration.

The results are given in tables 14 and 14a (Exp. 8). The results
for H«P0^ (table 14) are not very conclusive, as the rate of regeneration
(Fig. 18) is about the same for all concentrations which are not fatal. The

total amount regenerated is, however, greater in the three controls. The
neutral phosphate control lies midway between the two distilled water oontrols,

shoring that the presence of Na, K and P04 ions in the small amount employed is
neither beneficial nor detrimental.

The results of the NaOH series (table 14a) are definite, showing
a progressive decrease in both the rate of regeneration and the amount regener-
ated, as the concentration of base is increased. While the curve of growth

IB.
of tadpoles In 5 c.c. lies above the controls, the hydrogen ion determinations
•ho* that this eolution is not only neutralised but becomes 9lightly acid within
twenty-four hour*, §mi it h is already bean shown (Exporlurant 7) that this sane
coneentr-.tio ; of base is distinctly harmful if tha voluwa is sufficient to
pravent its bein; noutralizsi. Fig. 20 gives the time necessary to regenerate
25, 3" and 35$ of the amount removed.

The next series of experiments (9, 10, 11 ami 12) involving
124 tadpoles was made in order to compare the effects upon regeneration of
the various adds. Tha experiments were carried on in finger bowls containing

200 c.c. of the solution. Four tadpoles were placed ia each dish and the
solutions were ohangad rilteraate days. The acids used were 0.01 !f HNO3,
HBr and H2S04 end 0.01 raol. H3K>4# Tables 15, 16, 17 and 18 respectively

give the results of the experiments. The tadpoles in tUo HKO3 series were
•lightly •waller than those in the H2504 and HgP04 series, those in HBr sere
somewhat larger*

A comparison of the tables or figures (21, 22, 23 and 24) show
that In HKO3, H^r an! H2SO4 tie retardation of regeneration is gradual and
comparatively slight in concentrations up to 12.5 c.c. (.000625 H) . As the
concentrations rise above this, tha retardation to metabolism increases very
rapidly, the order of toxicity of the acids being HNO3, HBr, H3?04. This
might be explained on the ground that in the lower concentrations the acids
are so nearly completely ionised as to have about the smite effects, but as the
concentration is increased the concentration of hydrogen ions, and consequently
the toxicity increases mors rapidly in HNO than in HBr and in HBr than in
HgSOj. The data is, however, insufficient to «? arrant any conclusions. In

H3P04 (Fig. 24) there was no such marked retardation of regeneration in the
concentrations used although the decrease in the total amount regenerated wss
squally pronounced. : rate of regensration proceeded at very nearly the

19.
normal rate up to a certain point and w*s than discontinued abruptly. The
HgPC^ aerie* of experiment 8 (tsble 13, fig. 18) gave »imil%r characteristic
curves, but why the form of the HgP04 curve should differ from th*t of the other
eeide is not explained. Twelve an! one-half e.o. O.Ol Iff HSr or H2SO4 gives

about the ease initial hydrogen Ion concentration as twenty-five o.c. 0.01 mol.
H3PO4 beeauae of it a greater ionization, but by the end of twenty-four hount
the hydrogen ion concentration la -rue'; lower. This la beeauae the total
number of hylrogen ions whioh must be replaced before the solution will be
neutral is greater in the H3PO4 , consequently it is neutralized more slowly.
Thus the tadpoles |sj the stronger acids are in a medium of higher hydrogen ion
concentration each time the solution is changed but of lower hydrogen ion
oonoentration before it io changed again than those in tfta H3PO4, that lm, they
live under a more variable enviroutaorit. Further experiments till be needed
fcfj sho« whethsr this is the oauae of the difference in ths regeneration curves
or whethsr it is caused by some specific action of the HJPO^.

The tadpoles marked (T) (Tables 15, 16 and 1?) *ere transferred
to neutral distilled water to see whether normal regeneration could take plc-.ce
after the* initial stunting vith acids. Their subsequent regeneration is,
of course, not added in with the vtlMr tadpoles of th* group in determining
the p*r cent of regeneration. Tadpoles tranaferred luring the first days of
the experiment underwent more regeneration thar; those i«»ft in the solutions but
their regeneration was far below normal. At the close of the experiment all
survlvoro were transferred to distilled water but none of them underwent further
regeneration, showing that the effect of adds is not merely a retardation but
a permanent inhibition to regeneration.

The ourvee in figure 25 represent the time required to regenerate
?2.5£ of the amount retrove.1 in HHO3 and 25# of the amount removed in HBr,

20.
H2?-°4 snJ H3PO4. 7b8 sotted lines represent the aetual exp*rim*ntai data, tho

•olid lin^a the theoretlcnl curve. Excepting in the cnce of H3PO4, these linee
eoineile vary closely.

A comparison of the curves lor HHO3, H6r and H3S04 with the curves
for cirbon dio«ide production in bases (Fig. 9) for oxygen consumption In
bMM (Flft. 10) an! for regeneration in bases (Fig. 7) shows that they *re all
si'ailtr. Thess curves are also similar to ths metabolism cunrss drawn by
Powere (1919 b) from the data of Loeb and Kiss Hymen for oxygon consumption
in increasing amounts of anaesthetics. On the other hand, th-sy are ths

opposite of ths cunrss of Kro&h (1914 a and b) for Increase in ths rate of
dsvslopment at increased temperatures, ths toxicity curves of Powers (1918 b)
for ths decrease In length of life with increase in concentration of toxlo
substances and of the curve tir torn increase in regeneration in increasing amounts
of oxygon (F1&. 38) to be discuss at later. This would suggest th«t whether

criterion ohoaan be tine required to complete a given stage in development
(Krogh 1914 a and b), time required to ra^nerate a given p«r cent of the
amount removed, length of life (Powers 1913 a), or rata of oxygen metabolism, the
sffsot of temperature (Krogh 1914 a and b) hydrogen ion concent rati on,
insufficient oxygen >ni toxic substances (Powers 1918 b) follows ths same general
laws. That th**e various functions, - development, oxygen metabolism, regener-
ation, ems length of life - vary independently and bear no cattail relations
toward ona another is shorn by ths facts that an animal may be killed before it
has begun to regenerate in a ctadiut in which another may survive and undergo
conaiierable regeneration, while a marked change of 003 production may be pro-
duced by 1 medium which has apparently no effect upon regeneration. This
would indlests tamt tne threshold of toxicity of any given environmental agent,
as well as the velocity at which it booomej toxic#variea for the different
functions of the organism. However, that any environmental factor which

21.
affect* on* of tfceae funotiona will. If continued for •> sufficient time nnd
intensity affect ?*11 of thi function* eiitil^rly and in the euae direction la
eugge*tsd by the similarity of the curves. The only apparsnt exception Is the
increase in C02 production, due to conoontr.it ions of bases which o *u»oJ retarda-
tion of regeneration, an) issth. Mo«evor, an haa *e*n pointed out, thia

inore ao in oxygon astabolisft a<iy b« regarded as a destructive metabolism and
coap*r*blo to death rather than to growth or devolopswnt.

Figures 26, 27 and 38 are curves giving the tine neeasuir? to
complete the regeneration of various p^roentagaa of the amounts removed in
KKOg, HBr and H^BO^ respect ivsly. A comparison of these curves with the curvsa

for regeneration in bases of vari>.a« percentages of the amounts removed (Figures
12, 14, 16 ind 20) shows that while in sons cases (Figures 14 and 20) the regen-
eration of tadpolea In the higher concentrations of base* scorns to follow no
definite plan, that in gsnornl the curves are similar. In any given series
all of the curves of regeneration - which represent different stages in regener-
ation of the sans animals * are eonfconl. This shows thst while the rite of
regeneration at different stages la different, the p*r cent of retardation as
oomparad to th<» normal, due to tha acid or base,, la the asms for all stages, or
that for each concentration tha rei.tive velocities for the different stages
are prastically the seas. This is similar to the findings of Krogh (1914a)
in his work on the development of frogs' eggs at various tempo rat urea.

Ctrbon Dioxide Production and Regeneration in Acids.

The next eerlea (Fxrsriments 13 and 14) was undertaken to see
whether any correlation could be found between the regeneration and oarbon
dioxide production in acids. Ten sets of four tadpoles each of as nearly the
satis also as possible were use). The normal serbon dioxide production in

distillsd vatsr for n period of throe days was determined after a two-day fast

22.
to allo» tho aliaentary tract to become empty.

The first and tenth seta vers then kept in diatilied water as
control* and tvvo other sight put In 10, 20# 25 and 30 c.c. of HC1 -mi HBr K*de
up to 400 CO. with distilled water. Tho cnrbon lioxUe production, whether

expreeeed «• gra«s par dor or »» psr cont of norte%l, decreased progressively
with tho lncroaee in concentration of aoli, except Ir. case of the tadpolon in 30
o.e. of HC1 (T*!ble 19, Fig. 39) which showed a decided rlaa. The survivors

wore allowed to reganer-ita in the aatae concentr \tiona of <*olaa. Table 20 gives
the raaulta. Tha HC1 series ahow a progreseive ieoreaae in regeneration
(Fig. 30). Xu tha HBr sariaa tha tadpoles in 20tc.c. regenerated .-»ora rapidly
than thoaa in 10 c.o, after the seventh day. This ia probably attributable
to ehanee aa the tadpoles in 25 c.c. showed greatly -dec roaaad regeneration
and survived only eight lays. Figure 31 $lvoe tha rate of regeneration of the
HC1 serlee m time neeeeeary to regenerate 25, 30 ani 32.5^ of the amounts re-
moved, and the CO^ production of the aaeie tnllviduals ~.s c.c* H^COg par gran
of tadpole par d*y. It is provable th«*t for cent of aoretal COg production
(Fig. 29) ia tha worn logical tray of representing the effect of w environmental
factor upon netabolleie. Hare again (Fig. 31) the curves for various f9r cents

of regeneration are confooel, showing that far each concentration the relative
velocities of the d liferent stages are the sane*

Tables 21 and 22 give the results of an experiment In which CO^
detern. r*s were «ade Jaily throughout the period of regeneration. Four

rather larjre tadpoles i»ere used, care being taken to be** theti as nearly alike
as possible. The experiitent was begun at an alsoat constant teieperituro of
thirteen degrees centigrade, but later was placed at roow taatperature in order
to hasten regeneration. The effect of tno aoid in decreasing both the COj
production and the regeneration (T*ble 23, Fig. 33) ia pronounced.

33.
Figure 33 shows graphically tie average dnily C02 production, the
par cant of nonr.al CO,, production, the tot'il amount regenerated as per cant of
tho amount removed, and the tiaw required to regenerate 7.5jt of the amount re-
moved, while theee cxrvea aro in no wise cot3p'.«rable, they asnre to fcrin? out
more clearly tha progressive reduction in rate an! amount of raganeratlon and in
carbon dioxide pro 1 -action dua to increasing acidity of tha medium.

Halation of Site to Carbon Dioxide Pr-.-i ■action an.) Regeneration.

It haa already been mentions! in connection with experiments one
and two tnmt thara la a dlffaranca between large nnll tadpoles in tha

relation of retardation of regeneration »r»d length of life in various eonoentra-
tiona of baaea. Drayar and Walker (1914) show that in warm-blooded animals of the
aaite species, but cf dlffarent weights, dosage of drugs oust be Calculated in
relation to body surface. they explain this on the ground th<*t *the concentra-
tion in Imt plasma of any given substance administered la dependent on the
volume of tha circulating blood, which ia Itself proportion-vl to the body surface
in any given species of animal." Tha •basal boat production- of a wara-blooded
animal, which may be defined a« the heat proiuoel by the animal under consider-
ation when kept at tha same temperature aa ito normal body temperature at rest
and starving, ia alao regarded as proportional to body surface for any given

a pec lea.

A faw axporimente were undertaken to see whether any relationship

could be traced between the body are* or bodr weight of tadpoles ar.d the effects
of acids and bases upon regeneration pjai carbon dioxide production, which may
ba regarded '«s moat nearly representing the basal metabolism of tfef la.

Tor the firat experiment two tadpoles were taken which had been
oolleot»« at the eane tl«*,and iif fared enly in weight, git at care being taken
that they should be similar in ah^pa, relative langth of tail, sit. After
one week of starvation in distill*! trrtar/they were welshed aed the CO-

24.

produotion of each ^aa determined for three aucceaalve daya (Table 19) while
tha CO* production p*>r gram: .reight was not the aarae for the two epealnene on any

two iaye, the avomgee of tha thr*? d^ya 3.97 Mai 3.99 are prsaticslly identical.

It waa now hoped to determine th* ere* a of the t*ipol*e a* anaea-
thetiiing and making plaeter caeta of thaw, which eould later be narked off Into
a re f. a -it. J meaeurod »iih dividere. i» wetboi, however, proved unttatisf aetory

and waa abandoned.

For the next oxp«*rl*$nt (Id) a ne-w methoJ of calculating the area
waa deviaal. The average of tha length, width &ad depth of the hoJy wan
found and *lth I • the diameter tha area of tne body eae calculated aa the

area of ■ aphsre. Tho ares of the toll aaa computed by regarding it "a a
roo tangle fro* the baae to the level at which it begin* to taper rapidly to a
point, and aa a triangle frow thia level to the tip. Th«*ea MM* were doubled
to give the two alias of the tail, m -sres of the iv>dy, already found,

added to give tha total are** of I ' sole, whlc.i is axprceecd in atjuare

Millimeter*. Tha ore** c arvuteJ by thia awthod. i& probably considerably ieee
than the aotua.1 area of the iadpole, but it vn» hoped that by selecting eniaolc
eiailnr in ahape, Mm ratioa of computed area to actual area would be about the

Three seta of three tadpolen each were selected, &eaaured, weighed

"»nd Dm COo production deterwlnsd for six successive daya (Tnble 24). At the

end of thia time tha C0r fir l«» per gr»« of tadpole waa 4.65 c.c for the

2

larger else, 4.66 c.o. for tin aecond siae Med 5.0 o.c. for tha ewaller also,

showing a alight lnerets* in 00„ production par unit of weight, with B decrease

a

in else. In order to corapare the relative area, weight and CJ0£ production of
the three taJpolaa, all were expressed in terms of p>r cent of the larger also
taken ae 100 (Table 24). Tha weight and CO,, production vary b7 differences

of 4*1.13 % and +2.Q3 % in the aecond! and third alsea respectively, while the

25.
area and CO- production vary by differences of -8.8$ and -12.6$, suggesting that
the normal COo production of tadpoles in distilled water ia much more nearly
correlated with body weight than with area, as ie the caaa for <*arm-blooded
vertebrates.

The next series (Experiment 17) was carried out in the same
manner. Carbon iioxiia proiaction *as determined over a period of eight days.
The large tadpoles usad in thia axpsriment were much larger than those used
in the preceding, and it was noticed that the/ frequently exhausted the supply
of oxygen in lbs air in the experimental Jars. Thia was evident because when
the jars were opened, although the temperature had remained constant, the lids
came off with a "pop", showing that a partial vacuum had been formed. The
tadpoles also had a tendency to float at the surface, as they had been observed
to do in low oxygen water. For thi1? reason it was rather to be expected that
the C0« production of the largar si*e would be proportionately low, and this
ie found to be the case (Table 25). If now the large tadpoles be taken as
standard or 100$ in comparing the area, weight and C0« production of the three
■ets, the C03 production of the second and third sizes will appear abnormally
hirh. Thus the difference between weight and C0« production are *6.8$ and
♦4.2$ for the second and third sizes respectively, while the differences in
area ani CO^ production are -7.1 and -8.5. Despite the fast that the large
tadpoles in which COo production was abnormally low sere taken as standard,
the CO, production still corresponds more nearly to the weight than to the
volume. If, on the other hand, th? second size of tadpoles be taken as 100$,
the relative difference between weight and CO- production of tha third size is on-
ly -M. 6$, while the difference between araa and CO production is -10.7$, showing
again a very close correlation betveen weight and CO* production.

At th« close of the eight days in diatillsd *ater tue CO- production

m

26.

of the eame taipoloa woe Jit naift-ai for a p rlor. of six lays in 20 c.c. 0*01 S
HC1 made up to 400 o.o* - the same volume aa had been used in tw previous
sxperlaants. The C02 proiuotior; decreased in all three aata. Although tha
absolute deoreae* eae greater in th© l«rg>»r tadpol^e, the relative decrease was
greater in the smaller on?a, tha par cant dscreaeo Doing 33J&, 48$ and 5p£ in
tha largo, moiiuoi .tnvi small also* raapaotl vtly.

If tha decrease in COj production ha compared to weight and area,
uaing tno large else aa IV$> (Tabla 35), tha daoraaaa in COj production ia e#im
to be wore neerlj correlated with area than with volume. This experiment, while
suggostivo, la not conclusive, however, aa there were no distilled water oontrola
and aa the C03 production in distilled water showed a continuous decrease even
before the acli x?.a aided*

In the next eeriea (Experiment 18) the error due to the lirger
tadpoles exhauatlng the air in thnir Jar was avoided by putting the eane weight
rather than th« same number of tadpoles in eneh Jar. Tha Jere »ere divided
into four seta, each contelnin? three sixes of tadpoles numbered A, B, and C
(Table 26). The large tadpoles (A) were hardly comparable with the other
two slses, aa all had hind lege in which the sec on 4 Joint had already developed*
low. IA IBi 3A especially, were different, the forcer because it had a very
large compact body and short, stubby tail, the latter because the body *a*
rather long sr.1 fie l*gs were such farther developed than in any of the othyrs,
Tha individual tadpoles were weighed an:! tee as u red and the areas oomputsd as in
the above experiments. However, only the summaries are given hers (Tible ?S).
After the 00, production had been determined for six successive days in distilled
water, sat 1 was kept as a control, while eetj 2, 3 and 4, were put in 20 c.c.
of KOH, Ca(0H)2 an.l H3904 respectively, mode up to 400 c.c. Tha C03 pro-
duction in these solutions win* dstsrmin^d for four days. An examination of
the summary (Table 26) shows le the CO- pfSsfettiOB per gram varies from

37.
5.65 to 6.45 o.o. p*r Hy, even if ii exoluls the t*o inllviiuals (1A ant 3A)
slro*dy aentiosad as aberrant, taet this variation baera no apparent relation to
aise and thit the average COn proiuctljn la about the saws for all also*, thus
for also 8 it is 6.04 ani for alto C 5.96.

Duo to tna early death of soma of the tadpolea and abnora^l be-
havior of others during ths experiment, no attempt is stada to correlate the
inoraaari COg in baaaa or tha decreaeed C03 in H3SO4 'i** either weight or area.
Tha experlaunt doee she?, ho. ever, 1 14) par oent of increase of C02 in baaaa

and tha par cant of decroaeo of C02 In acids both increase aa tha *i»e of tha
tadpolas decraasa »n1, as tha relative are* of the tadpoles lnersaees ae tha
•isa deore»e:a, tfeia f«ot nay be regarded aa somewhat suggestive.

The surviving tadpoles of tha abova experlwsint *ere operated upon ani
allowed to un-argo regeneration in the earns concentrations aa tha C02 deterraina-
tlona h«J boon mede for. Tha results (Tabla 27) show that tha concentrations
uaod had no effect upon regeneration in eiae A. That elae B was affeotsd only
in tha ease of HUSO., in which regeneration waa noticeably retarded, •fell* In
thoea of size C which survived to regenerate, tlM regeneration *as aarkadly

depressed.

?or tha final aorlas on the affect of else on regeneration

srlaent 1?) 0.01 g HC1 waa uaad In concentrations of 30, 40, 50 and 60 o.o.
per liter. Tha experiments were carried out In 10-lnoh crystallizing dishes,
each initially oontoinln? tan tadpoles of turee different olzss. Two tadpoles
in tho first control and one in 40 c«c. XI Jutzped out during tha first few
days. All in 60 o.o. died without regeneration within ton days. An examina-
tion of the data (Table 28) shows thet under nor™ I conditions the average
per cent of regeneration Increaeas as the else of the tadpoles decreases. The
per cent of regeneration as compared to the controls in various concentration**
of BC1 ia practically tna sane for all. (The high per oent of regeneration,
as cuapared to the oontrola shown by the tadpoles of teodlun also, is lue to the

38.
abnormally low regeneration of the tadpoles of that also la the first oontrolV
A comparison of this experiment with Fx?*riment 18 (Table 27) sesms to suggest
that the depression of regenerat ion duo to acids la practic-lly the same for
sll alias of tadpoine until a concentration is reached which is soon fatal to
ths smaller onas. Tn such a concentration, those email ones which survive
to re<renar:»te, shoa • strong iepression of regeneration (Table 27). This would
indicate that the threshold of toxioity of ths lower animals is lower in relation
to the threshold of depression of metabolism than in the larger animals or, in
other *ords, that the effaet of the environment upon the regenerating tissue is
eomp-ratively independent ot its effect upon the tadpole as a whole. While
the data hers presented are both too meager and too inconclusive to warrant
an7 conclusions, these experiments and observations upon animals of various
sixes in ths previous experiments hr?ve led the author to regard this aa a
promising method o! a'-taok for future investigations upon the relation between
aew tissue and the organise as a whole.

Rsgeneration i& Low Oxygen.

The met nod of obtaining a gradient of oxygen has already been
described. For the first S3ries of experiments three tadpoles of different
sisee were placed in each of the seven experimental jars. Durinr this experimsnt
the water was strongly basic to Phonolpth-*lain though acid to Thytaolpthalein as
no attempt was made to acidulate the water. The data for the larger tadpoles,
70 to 77 ram. , are given in Table £9. The tadpole in the first Jar (oxygen free
uater) died without regeneration. Tha curve of growth (Fig. 34) of No. 3-in
1.6 o.o. O3 per liter - lies above those of 5 and 6 - in 3.03 and 3.8 •••« 02
per liter respectively. This is doubtless accounted for by the faot th«t
•I oa. 2 and 3 remain*! most of the time at the surface of the water, while
Nos. 4, 5, 6 and 7 regained at the bottom, earning to the top only occasionally.

Whan the faOt th*t the water in *hich Woe. 2 and 3 actually remained contained
cuneiderably wore oxygen than was shown by the titrations ia taken into consider-
ation, thaaa eunraa (Fig. 34) show clearly a relation between regeneration ani
the oxygen content of the r iter.

The aaaie thing is true of the medlua-aiaed tadpolea (TaMe 30,
Fig. 35). Of these Woe. 1 and 3 mala*! at the top and underwent regeneration.
Wo. 2, which did not show this rssponae, died without regeneration* Hoe. 4, 6
and 7, which remained at tho bottoa, undergo progra63ively better regane ration.
The dat* for the a:*all-ai*ed tadpoiea, 34 to 37 asm., are given .*t the bottom of
Table 30. Theee results are quite trpiosl of what occurred each tias an
attempt wee made to use 3**11 tadpoles in oxygen experiments. Woe. 1, 2 and

3 regained at the top of the *ater. Woa. 2 and 3 lived twelve drays and underwent
alight regeneration. 3os. 4, 5, 6 and 7 stayed at the bottom, and of these
only Wo. 7 survived to undergo any regeneration, but its regeneration wee
apparently normal.

Here again, as was thova for acids and bases, we find the suscepti-
bility of the tadpole as a whole as compared to the susceptibility of the
regenerating part ni$her in small 3 r tfeaa in larger tadpoles*

The next series of axperiraanta waa csrried on in water kept basic

to Rosalie acid and frooi acid to fcintly baaic to Phenolpthalsin. The teapers-

turs waa alao oorrectsd Ly the introduction of the air cell which »*armed the

water to air tempo ratura before it entered the e* nt J^ra. All of the

tadpole* choeen were large an J showed rudintent-'.ry hind legs, as it was found
that tadpoles at thtt stage ror-sal' I the bottom of the Jar* better than

thoae in earlier stages. The results (Table 31, Fig. 36) show an increase in
regeneration with an increase in Og content of the water with abrupt rises
bstweer. 0 an! 0.7 c.c. per liter, and again between 3.68 and 4.1 c.c. par liter.
the following experiment still oldsr tadpoles were ueed.

30.
The*? h~l h tot-1 length of 75 to SS go. and the hinl legs wore two-jointoi and
Ugited, and stood out (rorr. the body 00 *e to be vieible from the doreal aspect.
The raeults (Table 33, Figure 37) again oho* a oloee correaponlonoe u»t**en
the amount of oxygen and the rate of regenentic. Figure 38 le a curvw repre-

senting the length of time necessary to regenerate 20$ of the amount raaovad
In the increasing concentrations of oxygen. As has been mentioned before, this

-ve le similar to the curvee of Krogh (1914a) for Ihi iovelopiaent of frog
eggs at incr*-iain»; temperatures.

The final exporiaent of the oxygen eerlee owes Its v»lu* to an
aeeident. The tadpolsa of this eerlee were 71 to 76 mm. long *nd h/j rudimen-
tary hin lege. Bo. 3 died without regeneration. Of the other five, Woe. 1
and 2, In 0.2 end !•! 0.0. O2 per liter respectively showed marked retardation
of regeneration when neaeured on tho twenty-first day (Table 33, Fig. 39),
During the twenty-ascend alght, the floe of water through the jars stopped, due
to |l«ggiBg of t -ing apparatus by sediment from the eater, an*! before this

could be remedied the voter in the jars h&d beeosa w>sll asr&ted. During the
following night the cater again ceased to flow through the jure and again the
water in which tht* tmdyolma vera bce&sra aerate i. Since the tadpoles h»J been

two days in aerated *3tar, ih* original purpose of tho experiment h-»J to
be abanlonai. An aerating jar with a rapid stream of air passing through it
wax placed Just after ixing bottle eo thst trie water entered tho first

Jar aemtai. Measurements made on the twenty-sixth day of the experiment
ehoved that the tadpole in the first jar (0.2 c.c 0^ psr liter), which had
regenerated only 2.8j£ during the twenty-two days preceding the aeration of the
water, had regenerated 19.0£ in the four days following the change. The
tadpolsa already in hi ;her amounts of 02, of course, showed less difference.

I experiment is valuable as demonotr tin? th-st the decrease in regeneration

31.

in tha lower amounts of oxygen la due to the lack of oxygen and not to any Blight

rite in tamp^rature of the eater in the successive jars, or to a decrease in

alkalinity of the water as it took up carbon dioxide produoed by the successive

tadpoles.

It hae long been known that low temperatures, if not so low ?s to

produce detrimental effects, may retard or completely check the development of
certain ?ni*ale for a conslierable period of time, the animal still retaining
the power to complete normal development. The following experiment was planned
to test whether thia might be true for regeneration. Three seta of four tad-
poles each were seleoted. The first set was kept at room temperature - 19 to
21 degrees centigrade - the seoond was kept in a water bath at 14 decrees, and
the third was kept in a ref rigeretion tank at 0 to 4 degrees. The first set
underwent regeneration rapidly, the rate of the second set was considerably
retarded, the third set underwent no regeneration whatever (Table 34 and Fig. 40).

When the firet set had completed regeneration, two fror. the third set *»ere

were
trancferred to room temperature. The day in which they ^transfer red and aeveral

succeeding days were cool, so th9 temperature of the water stood at 18 to 19
degrees. For thia reason the regeneration of these tadpoles was slightly
slower than that of the controls, although the growth curves are very similar.
The two tadpoles left at the lower temperature died without regeneration at the
end of forty-seven days.

In comparing the 9ffocte upon metabolism of hydrogen ion concen-
tration, lo* oxygen and low temperature, it is of interest to note th t alike
they cause decrease and both the rate of regeneration and the total amount
regenerated. Were the effect of these agents due wholly to their action in
retarding the metabolism and consequent division and growth of the oells of the
regenerating parts, the result would not be a decrease in the ultimate amount
regenerated, but rather an increase in the length of time necessary to complete

33.
rogena ration. This, hi* ova r, is not the ease lor in acids and basee the tad-
pole* In the higher and moro detrimental concent rations oaaao regenerating at
a time as early or earlier than the controls. In low oxygen regeneration

ceases at practically tha aana tin? in all concentration*, while in low teaqpora-
tures, although tne period of growth la longer at tha lower tonne r-<-turas tha*
at tha hi^hor tomparss.turoe, the difference is not fUtfiftiftttt ti eotspenante the
difference la rate of regeneration, ao Ihs x&tlmate amount rogeneratei ie still
less at the lower than at the higher teiapmr^lur^s.

The only tonsbla explanation »ee?r»e to be Uut the phenow?r.a of
growth and of differentiation in tha regenerating pirta nr& differently af footed
by t'.e environmental agents studied, thr t ie, tfcftt tat optima for these
phenomena are different «o that a hydrogen ion concentration, an oxygen content
or « temperature of tha surrounding medium which retards ono process way hate
no effect wpon the other. Thus ft hydrogen ion concentration or deficiency
in oxygen which causae a narked retardation to regeneration ssay h^sve no effect
upon differentiation, while a temy * r it -*ra vmich retards regeneration &rsrttly
nay have only a slightly retarding effect upon differentiation.

That the three environmental factors etui lei may liffer this *ay
in regard to their relative effects upon regeneration ft«d differentiation seems
the mora probable since experiments show th«.t they do differ in their relative
effeots upon regeneration and survival or length of life, Thas a. concentration
of acids or bases or a deficiency of oxygen sufficient to completely chaoi

regeneration is rapidly fatal, whila a tadpole may live forty-eight lays at a
temperature whioh completely prevents regeneration.

33.
SUMMARY

A study haa bean made of the effects of various conditions of
hydrogen ion concentration, deficiency of oxygen and low temperature upon regen-
eration and metabolism in tadpoles of Rana olimata.

These studies show that the optimum hydrogen ion concentration
for regeneration is neutrality or very near neutrality. As the hydrogen ion
concentration varies from neutrality in the direotlon of either aoidity or
bacisity, both the rate of regeneration and the total amount regenerated decrease,
at first gradually and then very rapidly. These effects are due to the hydrogen
and hydroxyl ions.

The relative effect upon regeneration of any given hydrogen ion
concentration is practically the same for all stages of regeneration.

In water of low oxygen content, both the rate of regeneration and

the total amount regenerated are proportional to the oxygen present. Both the

rate of regeneration and the amount regenerated decrease with a decrease of

temperature.

Tadpoles, whose regeneration has been completely checked by a

decrease in temperature, retain the capacity to undergo a large degree of
regeneration to or beyond a psrlod at which control tadpoles have completed regen-
eration. Tadpoles, whose regeneration has been checked by insufficient oxygen,
•how this capacity to a lees degre3, while those whose regeneration has been
checked by acids or bases show it scarcely at all. It is suggested that this
may be due to a difference in the relative effects of the environmental factors
studied upon the phenomena of regeneration and differentiation.

Carbon dioxide production of tadpoles in neutral water is propor-
tional to their weight.

Carbon dioxide production is decreased by acids and increased
by bases. A high rato of carbon dioxide production, when produoed by a basic

34.

•edittK, 1* not correlated with a high rate of regenentlon. Both the relative

decrease of carbon dioxide production In acid* aui Ibi relative increase of
osrbon dioxide production in bases increase as the size of the tadpoles decrease.
This 1* »ugs»otiv* of ■ correlation bot»een the of foot of acids and bases and
the area of tM tiinole. The relative deereate in regeneration of tadpoles
dus ie aoida and basse appeare to be independent of eiae, escactin$ in concen-
trations which are soon fatal to the entailer ones.

The probability is suggested that hydrogen ion concentration,
insufficient oxy^et-, low temperatures and toxic substances affect devolopewnt,
regeneration, oxygen metabolism, and duration of life, according to the earns
laws, although the relative effeote of any given environmental factor upon the
variom functions of the organism Jiffsr, ami although the relative effects
of the same environmental factor nay differ in animals of different else*

This work was done In the Zoological Laboratory of the University
of Illinois, unler the 'ireotion of Dr. V. I. Shelf ord, to whoa, the writer ie
Indebted for n<%aj courtesies a:;.' valuable suggestions. The writer ie also
Indebted to Dr. Charles Zeleny for suggestions concerning the methods used in
the work on regeneration, end to Mr. F. D. Powers for many ueeful suggestions
concerning the details of the work and the interpretation of dot*.

35,

LITERATURE CITED

Bullot, o.

1904. On the Toxicity of Distilled Water (or Fresh Water Qammsruo.
Suppression of this ?o*i<5ity by the AMltion of S«all Quantities of
Sodium ohlorido. 9. of 0*1. Pub. Fnyelology, 1:139-141.

Chili, C. X.

1913. dies on ths Dynamics of Korphogeneeis ani Inheritance in
Experimental Reproduction. V. ?ho Relation Between Resistence to
Depressing Agsnta and Pate of Jtetatolieas in Planarla dortoettphalg, and
its Value as a Method of Invsstlgntlyn. Arch. Entw. Mech. Or%.t
14:153-?06.

Dreyer, George V. snd fa.lker, E. I.

1914. The Determination of the Minimal Lethal Dose of various Toxic
Substances and its Relationship to the Body Selght its Pant Blocdei
Aninsels, togsthar *lth Considerations bearing on tt;s Dosage of Drugs.

* Proc. Roy. ?oc. London, 87 B: 319-330.

Ha-rs, A. R.

1916. The Effect of the Addition of Alkali to Sea later upon the
Hydrogen Ion Concent ration. Jour. Biol. Chcp., 36:515-517.

Krogh, August.

1914a. On the Influence of Temps rH\rxr>i on the Rat* of Embryonic Development.
Zeit. All. Physiol., 16:163-167.

1914b. Ob U-s Rate of Development ami Carbon dioxide Production sf Chrysa-
lides of Tenstrio molitor at different Temperature*. Zeit. Allge«
Physiol., 16:173-190.

Levy, R. L., Reuntrss, L. 3. sjul Harriott, I. MoKim.

1915. A Slnpis Method for Determining Variations in the Hydrogen Ion
oentration of the Blood. Arch. Int. Vsd., 16:399-405.

Losb, J.

1898. fiber den Einfluss von Aikalien und Sftursn auf die Embryonals
Entwiekelung und das Tmehstum. Arch, Enter. Meoh. Org. 7:631-641.

1903. On ths Relative Toxicity of DlstiiUd K'ater, Sugar Solutions and
Solutions of -she Various Constituents of Res later for Marine Aniaals.
U. of 8*1. Pub., Physiology, 1:55-69.

1904. On the Influence of the Reaction of ths Ser- fmter on the Regeneration
and Growth of Tubulsrians. U. of Oml. Pub., Physiology, 1:139-141.

Loeb, J. end Yestneys, H.

1913a. Th'i Relative Ifluonoe of leak an! strong Bases upon ths R*t* of
Oxidation in the Unfertilised E£g* of ths Sea Urchin. Jour. Biol.
Chera. , 14:355-351.

36.

1913b. Tho Influence of Baeea upon tho Rate of Oxidation In FertilUed
Egge. Jour. Biol. Che»., 14x439*464.

UcCler.don, J. F.

1916. Hydrogen and Hydroxyl Ion Concent r-tt ion in Phyelolo^y and Medicine.
Medical Review of Reviews, 22:333-365.

Medea, Grace.

1919. A Study of the C^uaes and Extent of Variation In the Larvae of
Arbacln punotulst-. Jnl. Morphology, 30:317-432.

Moore, Bonjnmin, Fo*f, H. F., an 1 Whitley, K»

1905. On tho Effeota of Alkallee &tA tftldi - of Alkaline arJ Acid
lalta upon Browth Mil Cell Divielon in the Fertllieed Eg$e of Echinus
Proc. Hr*. ?oc. Load**, 776:102-136.

Pot* re, E. B.

1913a. A Collecting Bottle eepoctally adapted for the Cuantit&tiva and
Qualitative Deterainetion of Dieaolved Qaeaes, particularly vs.- 11
quantities of Oxygen. Bull. 111. State Lab. Vet. Hiet., 11:577-578.
1916b. The Goldfiah ( Bare » sine c;ira»aiue L. ) M a Teat Animal Ir. the
\y of Toxicity. 111. Biol. Moo., vol. 4, no. 2.

Shelford, 7. F.

1918. ffcptipment for maintaining a Flour of Oxygen free ^ater aaa* for
controlling Go* Content. Bull. 111. State Lab. Kat. Hiet., 11:573-572.

Shelford, V. . I Powers, E. E.

1915. An Fxporiwontil Stud - of ike Movent Nta of Herring Mil Other
Merino Fiehee. Biol. Bull* ,28:315-334.

fells, M. M.

Heaotio.is and Reeletonce of Fiehee i Lf Satur^l B&wircaatat to

tidily, Alkalinity nr.d Reutmlity. Biol. Bull*,29:231-S57.

Whitley, E,

1905. A Bote on the Effect cf Acid, Alkali an'. Certain Indicator* in
Arro3ting or oth«?«plee XfiflUMMll Dovelopwent of the £gga of

Plaur'jneot?3 ~l??.teeaa an* frchinue aaoulentua. Proc. Roy. Soc.

m, 1 n :1?7-149.

Fright, A. H.

1914. Worth Aaertcan *aur«, Wa4hington.

Zaleny, Charles.

1917. The Effect of Degree of Injury, Level of Cut and Time within
the Regenerative Cycle upon the Rate of Regeneration. Proc. Nat.
Acad, lei., 3:311-217.

3?

1 EXPERIMENT 1

Regeneration in Bases - KOIi

cc.KOU Length Length Length Regenerated in mn.
in 200 in mm. removed
cc.IIpO in mm.

date - be.«run 10/25/17-10/30 ll/l ll/? ll/'J ll/S 1l/"J
0 . . 1.2 2.1 2.2 2,8 3.4' 4.

^eegen. 8.4 14.7 15.4 10. G 2'£, 8 20.4

1/4 53 12.0 1.3 2.0 2.5 2.0 3.1 3.7
''~egen. 10.8 16.8 20.8 24. 20. 31.

1/° 54 32.5 1.1 2.1 2.5 2.0 3.3 4.0

^rep;en. 8.8 10.8 20. 23. 20.4 32.

2' 55.7 12,9 1.1 2.1 2.2 2.5 3.', 4.2

^regenerated 8.5 16.3 17. 10.4 2r>. 32;5

5 57 13.3 1.0 2.1 2.6 3.0 3.7 4,4

^regen. 7.5 15.8 19.5 22.0 2« . 33.

10 5C 12.0 0.8 1.0 2,0 2*5 3,1 4.0

^rcgen. 6.6 15.8 16.8 iU.4 21.3 33.3

15 59 13.8 0.8 1.2 1.3 2.0 2.8 3.3

Jfregen. 5.8 8.7 0,4 14. > ':>. 24.

20 5(2 12.9 0.2 0,8 1.1 1.7 2.0 2.6

^regen. 1.5 6.2 8.5 13.2 tfcVS 20.

25 55.5 12,1 died without regeneration

Date TT/9 TT/12 li/l4 fl/lE 71/20 il/22 IT/24

0 1 4.6 5.1

Veg. 32. 36.

1/4 4.7 5.0

'reg. 30. 41.

1/2 4.3 4.8

#reg. 34.4 38.

2 433 5.0

#reg. 33.3 39.

5 4.6 4.6

#reg. 34.0 34.0 34.6

10 4,4 4.-0

#reg. 36.6 39.

15 3.5 3.6

^reg. 25. 26.

20 3,1 3.3

#reg. 24. 25.0 25.6

A.

5.6

5.8

6.4

6.2

6.°

»-^r» .

40 .5

45.

43.5

43.:.

5.1

5.4

. 5.8

5.8

0

•

'42.5

T>.

4T„

47.

t?«

4.8

5.0

5.5

5.5

38.

40.

44.

44.

42.

5.1

5.3

5.3

5.4

5.3

29.5

41.

41.

41.

41.

4.6

4.7

4.8

5.1

5.1

34.6

Or O

;;6.

38. •

^^ . 1

4..0

4.0

4.0

4.6

4.0

39 .

,

3.6

3.8

dead

26.

27.5

3.3

dead

TM3LF 3 EXP^RIIFHT 1

Regeneration in Ba9as KOH

cc XOH Length Length Length Regenerated in mm.
in 200 in mm» removed
oc.H20 in mm,

dafe - begun 10/25/17 - 10/30 11/1 11/2 11/3 11/5 11/7 11/9

0 ^ 23.7

5.3

1.3

2.

3.3

4.6

2.9

3.0

3.2

/regenerated

23.

40.

44.

*$..

ser-
vo .

56.5

60.

* s £6-

6.1

1.3

1.9

2,0

1,2

2.6

3.6

2.9

^regenerated

31.

31.

33.

34.

42. 6

43.6

48.

1 f 87

6.1

1,4

a .o

.-

3.7

3.0

3.1

3.1

generated

23.

33.

39.

44.

49.

51.

51.

2 ^ 26

5.

1.1

2.

2.3

£.6

3.6

3.7

^regenerated

20.4

37.

37,

37.

44.

44.

46.

5 28.5

5.

i.3

2,0

2.0

3.3

3.5

2.7

2.7

v venerated

Si

38. S

38.5

43.

43.

53.

53.

10 t m6.5

5.5

1.0

1.5

1.5

3.0

2.0

2.1

3.3

^regenerated

^13.2

27. 3

37.3

36.4

36.4

38.

40.

15 26.

4.8

0.6

0.9

0.9

1.1

1.3

1.6

1.8

1 generated

13.5

18.7

18.7

23.

37.

33.3

37.5

20 28.

6.0

dieu without regeneration

•

continued from above

K0H date 11 /l

8 31/14 11/1

7 ii/;

20 11/23

0 3.2

3.3

3.3

3.3

3.

2

£regen . 60 .

62.

62.

90.

60.

i 2.9

2.9

3.0

3.0

9.

0

^regen. 43.

46.

49.

49.

49.

1 3.1

3.1

3.2

3.3

3.

3

^regen. r31 .

52.

53,

53.

53.

2 2.7

2,7

3.0

3.0

ds

tad

^regen. 46.

46.

51.

51.

5 2.7

3.7

3.7

dead

#regen. 52.

52.,,

10 8*2

dead

40.

15 1.0

lead

$regen. 30. 8

TABLF 3 FXPFRIMFHT 2
Regeneration in Baaes - Ca(0H)2

No, Ca(0H)o.t*ngtb btaftli Length P.. rate* in mm,

0~in mm. removed

.

.

itf

-te

- "be run

10/ 30/1 "3

' 11/4 11/9

11/1

18 11/14 11/1

7

1

0

11,1

u

'.

3,

•0

4.7

5,0

<

-.

v.

34.

41.

43.

V

0

45

11, •

1.0

3.3

3.4

4.5

4.

4.

%

.

38.

31,

41.

.

1

1

47

.0

,

3.0

3.

4.3

4.5

4.7

9.0

.

31.

37.2

37.

.1

3

i

40

13 a

A. . JL

4.1

o.3

5.3

6.1

0

8.4

.

30.3

41,

•k.

46,5

0

a

4

1

45

ii.i

0.?

.

3.

4.3

■ i

v *

k.3

r*

i

.

.

.

.

■

a

5

3

4S

12. i

.

.

♦ .1

,

5.1

3. 1

r*

$

8,3

<» 0 ,

*.

41. G

.

i (V . *2

6

5

<*■*

11,3

0 . 0

3.0

3.

4.8

4. a

4.8

%

,

3w>,

40.7

.3

43.5

7

7.5

49

11*0

*,<s

.

9.

4, '

4.0

4»C

r

$

6,8

20.

SM^t

3«,

■

I,

8

10

45

IS.

.

.

4.8

4.4

4.

1

i

,

so. a

8*,8

.37,4

,

-

1

14

47.

U.O

.

.0

3.1

3.1

3.3

;•

1.8

18.4

>4 •

*

.

36 .

1C

I7. 1

43

d

11,

• —

18.

.4
31 , *

I.I
3&.

.a

do!^i

11

£0

48

10.

'

! without re-^nu ration.

So.

Ca(0H)3

date i:

L/M

11/

a/27

11/

1

0

5,

.

.

1

5.5

5.3

*

46,

>

.

51.

31.

I»

0

4

► 7

5.0

5.0

•

45.

4a.

t

5,

► 0

5.

,

1.0

;

41,

.

41.

0

U.G

41.

3

«

6,

,5

8.

5

ft. .

8.8

;'

49:

i

4*'.

V

1.9.

fcf<

4

1

4,

>4

4.

8'

»

.

39

.

43.

45. 45.

5

2

.

.3

.

7

5.7

5.7

H,

>

47.

47,5 47.0

6

5

f

5

44,

»

44.

0

ad

7

7.

4,

4.

'3

.

.

*

4'5,

>

4:5.

44. a *

.

8

10

4,

► 3

4.

4

.

.

:

3?,

i

37.

41.

U.

9

>5«

,1

3.

1

".

.

*

•

ra.

.

51.6

10

40

TABLF 4 FX^RrrFNT 3

Regeneration in Banes - Ca(0H)3.

Mo. Ci(0H)2 Length Lsn-rth Length Reranerntei in wai.

in .£00 in arc. rsariove-l
co.H^O in ma.

date - oe/un 10/30/1? 11/4 11/7 ll/9 11/1^ 11/14 11/17

1

0

31

7.0

1.1

'J. 3

.

,«

-j , 8

,

,■

16.

13.

40.

40.

43.

i"

0

31

6.9

1.

..

1.6

1,

.

o

;-

13.

33.

37.

17.

40.

3

3

J

1

7.0

1.

.

1.7

.0

1.3

.

CH

t

17.

91 rt

,4

41.

47.

47.

a

3

i

..

2,

,

.

1.0

3,0

.

► L

t

16.

.1

40.

47.

47.6

47.

4

1

SI

6.6

1.0

.

■

i

1.5

.

vr

t

15.

.3

.4

,

11.4

o

5

31

7.1

X.

.1

8*6

3.0

1.

3.1

0

i

%

17.

,C>

36.6

**•

46.5

,

^

6

5

30.

7,0

1.1

'•3.4

3.0

' .

-.

1.5

,4

^.3

43.

46.

16.

.

7

7. 9

6.

■ •

.

9 •

.

3. 1

2..-; dead

i

.7

.4

H*

35.

44.

41.2

B

1C

. .

1.0

1.9

51

•

37 .

'

9

15

6.1

.

,9

8,

..

a
•

t

14. |

30.

32.

,

10

17.5

U

i-

7.1

0.7
10.

1,1

ll.7

1.6

33 » 0

je&d

1

34

6. 5

0,

<4

4#6

Ho. Ca(0H)3 AftW 11/19 11/22 11/3?

1

0

*

.

1.0

-4

41.

.

.

V

0

.9

2.

.

*

J.

U.

42.

2

i

3.1

.1

$

47.

-.

47.

3

I

3.0

1.0

.

t

47. G

47.

47.

4

1

.7

.7

di

t

41.

41.

3

2.

3.1

i.

+

.

.

.

6

6

.5

.

.

i

50.

5',) .

50.

41

TABLE 5 SXPSRIM5OT 3

Regeneration in Base* - Na^Ctt^

.01 isol,

1*2 CO3 Tad. Length Length Length Regenerated in naa.

in 300 no. in mia. removed

OC.H^O

Be.-un 3/27/18
0 1

in ma.
11.8

4/3

. &

4/4
3.4

4/6
5.

4/8
6.0

4/10

7.0

4/14
7.5

4/18

5.3

4/33

5.9

4/34

4/38
5.9

s

50

13.3

.5

4.0

3.8

4.4

4.9

5.0

8.0

8.4

8.5

8.5

3

49

11.

3.3

3,1

M

4.7

.0

5.6

6.0

6.0

6.0

6.0

4

total

44

13. (

§.0

>•

lTTT

4.2
1^77

B.C

affTT

5.S

3*1

3TT

5.6

6.0

TJS. 3

6.0

387*

6.0

.4

i

18.

38.

34.

41.

45.

48.5

51.

54.

54.

54.

15

1

47

13.1

1.6

3.3

3.8

5,0

4.9

5.1

5.5

5.7

5.7

5.7

a

49

.0

1.5

3. £j

3.5

4.4

5.0

5.5

5.7

6.0

6.1

6.1

3

45

11.8

.0

3.1

4.0

4.5

5.0

5.6

5.5

5.0

5.0

5.0

4

Total

44

TOTS"

77T

3. j
1 f>. 3

3,5

iTTS"

lCT

4.7

i~7£

5.4
8l7f

5.6
8 «v . 3

6.C

3377

6.0
. 0

5.6

3 3, '4

i

14.9

35.3

30. I

37.

40,

44.

46.

46.5

46.7

46.

3C

1

43

13.5

1.3

1.7

2,

. 3

4j

3.9

3.3

4.0

4.0

4.0

a

50

12.5

1.7

3.1

3.0

3.7

3.3

8*3

4,3

5.0

5.1

5.1

3

45

11.3

1.7

3.7

3.4

3,1

3.1

5.0

3. 1

3. w

3.9

4.0

4

43

U.I

1.3

3.S

3.3

3.7

a. a

4.7

5.0

5.0

£>.

3.8

total

tsttt

672T

S3

lTTS"

ITPZ

1375"

iunr

lTTC

1773*

187ST

1ST*

I

13.4

19.3

34.4

36.4

«*9.

35.

35.

$7.

5 ' , 0

34.

45

1

5*

U.l

1.1

1.1

1.7

3.0

3.3

3.3

3.1

dead

2

48

13.0

1.0

X , A

1.8

1.6

1.6

1.8

1.8

dead

3

47

11.9

0.6

1.8

3,4

dead

?•¥

4
total

40

LLC
Z57~

l.C

377

1.3
»77

1.6

777

lead
373*

T7T

TTT

15.5

<*
1

7.7

13.6

16.

15.1

16.3

16.3

60

died without regeneration

•

1

75

5.9

6.0

6.1

6.6

0

1

5"

11.5

3.6

3.1

3.5

4.3

4.3

5.3

5.4

6.5

S.5

6,0

48

12 . 9

3.3

3.3

% . ^

4.0

5.1

4.3

6.1

Ll

6.0

8.4

3

43

11.6

, •

3,3

3.'

Tt » 4.

4.8

5.1

6.1

« ••

6,7

;.3

£.t

1

44

13.2

hi

37T

>»a

4.C

4.5

s.a

5.7

wS79

aTJTT

3*73"

frot ■..:

TECS'

x -.0

l^T

lSTST

1377

aSTT

47.5

51.5

53.5

50.

?

19.5

36.6

3:.

35 »

40.3

43.

Approximate PH
initial after 43 hra.
7 6.8

lOf

7.4

7.8

6.8

/

42

TABLF 5 a EXPFRIM^NT 3
Regeneration in Baeee MaflCOj

•01 sol,
NaHCO,

in aoo

oo.H20

0
15

Length Regenerated in mm.

H

H

45

75

90

120

Tad. Length Length
no. In bub, removed

Begun 3/37/18 4/3 4/4 4/6

controls the eame ae tot NasCO^
1 48 12.6 3.0 3.3 4.

2

4
total

1

a

3

4
total

1
3
3
4

total
I

1
3
3

1

3
3

4
totfbl

r

i

2
3
4

total
i
1
2
3
4
total

49
48
49

48
52
45
45

53
47

43
51

Si
52
46
41

55
50
45
43

54
49
47
44

51
51

ii.8
45

11.8

TO"

13.5

13.0

13.0

1.6

T57T
12.6

ti.a

11.4

...

4"0*

13.6

I .
IX. 5
16.5

SETS'

13,1

13.0
11.3

11.8

T377

12.7
12.3

13.0
11.3

TT"

12.0
11.9
11.8

1 ..

-77~

.3 4.0
1.8

1.7

77T" lTHT

16.1 26.5

3.0
2.1
3.1

1.3
^7T
16.5
3.0
2.1
1.9
\.

777

16.4

3.0
3.1
3.1
1.6

16.

1.3
1.8
3.1

...
77T
16.
3.1
2.0
1.8
1.7

77*r

15.4
1.9

3.0
3.0
3.0

3.0
3.1
3.1

3.6

iTTo**

35.
3.3
3.7

2.8
3.6

lTTT
23.6

3.4
3.0
3.5

lTTST

24.3

3.3
3.1
3.~

1T7**
23.6
3.3
3.8
3.1
3.3

lO*

34.
3.5
3.0
3.6
3.3

.6.5 IB ■

ie:

4.1
3.4
3.3

llSTC

31.

3.7

I . I ■•.
3.6

3.3

lTTT

39.
3.6
3.8
3.4
3.4

lT*T

29.4

4.1
3.9
3.7

ia

1TTS"

30.4

3.3

3,3
3.5

3.5

13TS

37.6

3.3

4.1

3.3

3.1

lSTS*

38.

3.0

3.1

3.0

8.7

4/8 4/10 4/14 4/ia 4/33 4/24 4/38

4.3
5.7
4.4
3.0

3375*

98. 3

4.1
4.1

3.9

4.0
lTTTT
33.4

4.1

4.3

4.0

3.8

i"CT

33.6

4.1
4.8

4.3

4.0

lTTT

35.

3.3

4.0
4.1
4.1

1BT5*

31.5

4.0b

4.7

3.9

3.8

iSTT

33.

3.1
3.6
3.1
2.3

4.9
6.4
5.0

tint

39.5

4.3
4.5

4.0

4.2

lCT*

35.
4.6

5.0

4.2

lTX
37.

4.3
5.3
4.6

■:.{.
1S77
38.

3.9
4.6
4.6
4.1

lTTT

35.
5.0
5.0
4.5

.:»..

18T3"

37.
3.3
3.6

3.2
3.0

5.3

0.9

.
3.2

■TTT

43.5

4.9

4.8
4.5

*7T"

38.
5.3
4.6
4.8

6.0 6,0

8.0 8.0
5.7 6.0

mlealng

1377 2oT~
51. 54.5

4.9
4.3
4.5

i.g

iTTTT

39,
5.6
5.0

4.9
1F?T

leeing44.
1477

49,

5.8
8.0
4.8

5.0

44.

4.2
5.2

4.9
5.1

157T
N . 5

5.6
5.7
4.8
4.3

257?

41.
3.3
4.0
3.5

3.C

:-~7

5.7

6. m
5.2

33TT
45,

6.0
4,9
5.1
mi* a
17575*

43.

5.8

6.1
5,5

4.9

sSTT

45.
4.1
3.0
3.3
3.3
1378"
38.5

5.0
5.1
5.1

4 . L>

1*377

40,7
5.5

5.0

5.0

1*575*

44.

6.2
5.7

5.^

5,0

45.

8,0
5.0
5,1

43.
5.6
6,1
5.5

4.9

?JZ7T

45.
3.3
4.1
3.5
3.1

14*7B*

wis .

6.0
8.0
6.1

soTT

55.

5.0
5.1
5,1
4.5

1S77

40.7
5.5
5.3
5.0

1373"

45.

6.3
5,7

5,3

5,0

.TT
45,5

6,0
5.3
5.1

lO*

43.5
5.8
6.1
5.3

2T7T

45.
3.3
4.3
3.5
3.3
TT

39.7

6.0
3.0
6.1

3o7T
55.

5.0
5.1

5.0
4.5

157?

40.6
5.5
5.3
5.0

1*573"

45.

5.7
6#3

5.3
^6,0
32 , ii
45.5

8.0
5,3
5.1

1*37?

43.5
5.8
6.0
5.3

*.■?

.^7~

45.
3.3
4.3
3.5

.

29.7

Approximate PH
Initial after 48 hre.
7.7 7.3

7.9

7.5

8.3

7.8

8.1

7.7

1.8

7.9

8.6

8.7

8.1

TABLE 6 EX^FRIIFNT 3

4

Regeneration in Bases - N&2CO3 + N&HCO3

in 200 in 200
CO.H3O 0C.H3O

15

45

Tad* Length Length

no* in mm* removed

in sue*

begun 3/27/18

1 50

Length Regenerated in bob.

2 54

3 43

4 47
total

t

1 49

a 47

3 44

4 45

total

12.5
12.5
11. 6
12.1

*S77

11.8
12.0
11.6

1.'.

4TT7T

4/3

2.0
2.1

2.1

'.- . "
18.

1.8
2.0
2.1

2.0

773*

4/4

3.1
3.0
3.8
3.8

137T
28.

2.5
2.9
2.8

V.

77"*

4/6
3.6

3.8
4.2

4.

io77
33.

3.0
3.3

3.1

3.3

iJCT

4/8
4.1

dead

6.0

5.0

i*CT

38.

3.6
3.9

3.6

3,7

l^Tcf

4/10
4.6

5.3

5.1

15*7o~

40.5

3.9

4.0
3.8

4.3

i*CT

4/14
5,3

■ «

17. 04.2 37.5 32.4 34.3

3.9
5.0
4.5
4.0

37,5

4/18
5.8

4/32
6.0

4/24
6.1

4,1

5.5

V>

...

1377

41.

4.1
4.9
4.4

.....

41,5

4/38
6.1

6,6 6.1 6.1

5.9 -. 6,0

~ iTTe isTb" 18T iK?

44. 48. 50. 50.

4.1 4.1

4.3 4.9

4.4 4.4

.i~ ifxr

41.3 41.5

Approximate PH
initial after 48 hiv>.
7 6.8

.'•'

M

30

1 til

2 53

3 44

4 50
total

13,0
13.5
11.4
11.8

1.4
1.1
1.0
1.6
57T

2.0

2.3

..:
wTTT

2.7
2.7
2.8

2 . 3

lCTT

3.0

3,0
£.3

ir.r

3.1

2.8
3.0

2.5

lTTT

10.3 17,5 31.

32,5 -3.

3.5
3.1

.;~7T

24.6

3.6
3.2
3.5
3.1

13TT

37.

3,6

3.0

:.:,',,

3.6

3,7

4.0

iTTT

3,6
3.7

4,0

-

30.

10«

7..

45

15

50

25

16

BO

1 47

2 53

3 43

4 45
total

I

1 5.

a 49

3 46

4 46

total

%

1 53

a 51
3 44

4 46

total

11,8
11.6
11.5

■ .
47.7

12.8
11. 5

13.0
11.8

i.t.O

11.6

12.8
11.7

77

3.1
1.0

...
TTS*

Q.t

173
5.3

1.7

1.6
1.5
1.3

o

1.1

1.3

1.

2.C

5.5

1.6 1.4

1.7 3.1
1.3 1.8

7*77 775"

8.2 11.5 14. 15.7 15.7

<;ead
lead
'<ead

0.9
0,9
7.6

3,1
3.5

1.1 :,*
TJL 173
9.34 11.0

2.4

.. i
3.1
3.4

l""7T

3.7
3.0

1.7 dead
dead

2.0 dead
dead
377

dead

2.9
3.7
3.0

3,1

3.7

3,7

3.7

3.7

3.7

3.3

3.4

3.3

|M ■

3,2

3.8

3,7

.i.8

3,8

*7c

lsfr

3.4
14TT

lT^T

int?r

13.1 19. M 26. 36.2

29.3 39.5 39.5 30.

I '

TABLF 7 EXPFRI1TOIT
Carbon iioxl.i Proluotion In Bnaea - rOH.

Ko,

, ooKOH
in 400
-•.KtjO

CO- ■** 3C. 0.0 IN Ho

S03

data - •

- 1/17 1/1G 1/20 l/;il 1/38 1/33 1/34 1/25

1/26

1

0

3G. .7.7 44.6 41. C .3 .9 36.3 98. 1

44.5

15

37.5 41.3

1.7 44.7 44.9 43.0

43.0

3

25

44.0 47.5 *5.5 49, . 60.6 , 52.0

61.0

4

30

49.7 55.

5

35

48.6 ltad

6

40

51.

1/31 3/2

Ho.

KOH dat

1/30 3/1 ,3 a/4 i/i

I 8/6

1

0

tt.3 48. a 37,0

33.0 43*6 33.5 37,6 ,7

,4 37.0

31.7 51.3 43,0

37,5 53. 5 11.3 33.7

. 1 30 .

3

43.4 70.0 48.

48.7 63. S 87.0 41,0 41,
wt.in total

,6 43.3
CO3 per

No.

KOH d*t<

,'U gmB» days CO3

day

1

0

3,6 27.;- 39,6

20.8 6. ^4 799.5

33.3

15

•'.1 39 • . .

19 10 6,9 34 863.9

36.0

3

37.0 3S,0 39.7

37,5 7. 34 1038.4

43.

No,

KOH

CO2 per day ^r*«.

1

0

4.03

a

1

5.

3

. .1

16

Tmr 8 EXPFwrr

Be generation In &%*«• - KOH.

Ho. . JH Tv.i. Lon

In 400 no. in aft.

oo.H30

Length Length Helens r^toi in mm.
in taw.

X'J m

1/17/18

1/

3/

3/7

.'13

'17

8/38

0

1

71

u

4.

,7

.?

r,

7.

7.4

2

3 .

'.

•

i.a

.8

3

N

11.1

:i

* 4

J2l

1.3

■ •

3.1

>■' •

» -

3.8

Tot>*l"

■

T~7T

iTT"

1 *• ■—^™"fc

i37""~

._

I

.

47.6

51. Q

54.4

• 9

1 .9

16

1

13

Lftl

3

.

.

1

.a

i.a

■

t.

3

11.

,

•

7.

7.

-

7.0

4

LO

■

'TT

.-.

177T

■■..-.

Tot, a

3TT4

6?t

1773*

177"

177T

£

r»j

41.

60.

so.

^•S

1

74

i *

M

A . '.;

'l •

3.

3.

1

••

-.1

.3

4.1

3.

ie*A

3

in.

.

9.

3.0

3.

It

4

'

10

8,7

J-7

..

TotaJI

Tb

77T

;"T£

1^7?

T7"

377

i

16,

.

39.

.7

30.

30.

• T>ii« taipol*? #aa abnoiv >th in btfn&vler M I ir. fora of

the \ trt so li not indu-lcV in the total.

16
TABL* 9 FXPFPIIFNT 5

Carbon dloxid production in Base© - KOH

1 3 3 4 & 6 7

Wt.of 5

ttvipolee -4.7 5.1 5.0 5*0 4.9 4,6 4,6

Normal CO-. Production in Distilled Water

date

3/9/18 29,4 34.7 V3.9 31.9 34.4 30. 34.5

10 7.1 33.7 30.3 ,5 31. 23.1 SB.

3/11 . . >.B 30.7 .5 •14.15 26.

3/1 .. 34.1 . . 34. , 34.1

3/13 . .4 .7 33.3 30.3 14. ,4

3/14 a > -v.) •■■ . .1 . v

Total CO 163.4 177.3 175,1 .9 175.7 11). 1 164.4

A*. Daily 37.08 .65 .18 30. ,3* 18. 37.4

CO.

COj per

g*.day 5.76 .31 .83 6.09 5. 4.0 5.

CO^ Production in KOH

oc.KOH in

400 00 iro 10 15 20 «5 30 M 0

3/16 ;.l 38.4 30.4 33. 38.9 44.0 19.8

3/16 31.8 . 4.4 . 31.0 33.9 14.7

3/17 26.1 . 34.1 3C. 30,8 .iea.1 ..0

3/18 . ,8 31. 33.3 dead 18.

3/19 :.. ■,.;> -.7 31.4 ___ __ 17.4

Total C03 i.3 .2 .6 i.l .7 77.9 ,D

CO,, per day ^4.84 .34 . , 38.95 18.3

C02 par

(■•day . 5,46 8. 6. 6.85 8.46 4,0

i» of Normal

1. 93,9 10'-. . 114.7 . 66.8

• The Ba(OR).; u*el to abaorb the CO^ became mixed with the water
When fount the ta'polea were almoet deal ani frojt that time
they *ere abnormal.

oo» KOI

In 30C
oo.H^C

15

■0

88

TABLr 10 EXPTSIMTfflf 5
Regeneration In Ba»ea - KOH

47

00 i

KOH Tft .

Lenrth

Length

Lanrrth Re;!;

ane rated

in

300 no.

in m«

removed

00i

Hw0

in raja.

be^un 3/19/18

?/

3,

3/30

4/3

4/5 4/9

4/13

4/17

4/.31

1-

1

43

11,2

1.1

. -

3.5

5.0

5,0

8.0 7,0

7,0

7,0

a

45

9.8

1.6

.0

.9

3.

4.0

4.3 4,7

5.3

5,1

.,.

3

44

10.

1.

1.8

2.6

3.7

4.5

4,5 5,0

4,9

4,9

4.8

4

totn

45

xtfr

3.Q

~T"

lOT

.ittit

lffTTT T77

4,f

5.0

I

10.4

19.

29.0

38.

48,

46,

54,

• 5

55.7

l

51

9.8

1.0

1.4

2.1

»J ,9

3.3

3,5 5,0

5.0

5.0

5.0

2

48

11.1

0.9

1.6

.1

3.8

3.7

5,1 3,5

3,5

3.1

3,1

50

.J

1,1

»0

,«

%*

4,n

4,t> 5.1

5,1

5,0

5.0

4

46

11.1

0.8

1,8

3.4

3.9

4.1

4,2 •

3,3

.

3.3

5

total

44

...

TH?

£.0
TfTBT

i

irhr

iTTT"

lTTTT

4*5

scTT

4.7

T7~

4.5

~7"f

1 •

?

9.4

16.8

23.

33,

36,

39, 41,

41.

40,

40.

30

1

48

10.1

0.

1.6

1*0

3,6

3.6

5,0 ->.

4,9

5,1

.

49

11.0

0.7

1.3

3.3

3.5

3,8

4,3 3,0

5.

3.

4,0

3

48

11.

1.0

1.8

3.8

3,0

4,6 4.1

3.9

4.

4.

4

10.

.

1.3

2,3

3,1

3.3

3.6 -■■,

4.3

dead

5
total

4C

KT

1.7
777

iTnr

iFTT

l77T

4.3 4.8

'■'•1, 7 •

4.8

lT7~

*

S.5

14.4

23,1

26.

■'' .

40,

43,

40.8

40,8

25

1

46

10.0

0.5

1.7

.8

3.

3.3

3.6 3,7

3,7

dead

3

48

11.

X.

1.5

.1

2,

3,3

3,8

3,3

dead

3

46

10.

1.0

1.3

1.9

.5

3.3

3.S 4,4

4,5

4.

Lead

4

total

vrrr

0,

?7T

1.6

2,0

lTTTT

i"v7"f

'••• •' 3.3
l"OT l^TS

3,4

lTTT

V| .

"T"

*

7.4

14.

•1,

.

31.

34,

38,

37lii

.

0

1

49

10.3

1.0

.

.7

.4,0

-4.6

.5,4 4,9

5,9

5.5

5,6

a

47

11.0

1.1

2*0

2,8

3,7

4.4

4.8 5,3

5,3

5,7

5,5

3

44

10.1

1.1

1.8

.7

3.6

4,0

5,0 5.0

5,0

5,0

4,6

4

49

10. 8

1.0

1.6

2 •

3.9

4.

4,6 4,4

4,8

4,8

4.1

5
tot=*l

i

47

7*7T

•

9.9

—

18.

35,

itW

36.

41.5

4,3 ...
33.9 24.8

45. 4?;

II:8

4,0

35,0
47.

...
47.

Approximate PH
Initial .after 48 hrs.
9 .7

3.3

'..

1C

.-

6.7

48

TABLr 11 ^TPJMTHT 6

Carbon dioxid Production In Baaee - Ca(OK)£

Ko. 12 3 4 5 6 7

Wt.of

Tadpoles 4 4,1 4 4. 4 3.8 3,7

Normal CO., Production In Distilled Vater
vte

29.8 .7 .4 •17, .3 38.7 28. S

3/1 . 33.0 36.8 .5 .7 >.8 .4

3/11 33.7 33. 9 <;2.7 17.1 . .4 22.1

3/. 21.3 23.3 31.4 16,1 19, . 18*4

3/13 , .2 , 1,0 34. 35.0 17. b

3/14 . 21.7 ^4.7 19j |3

Total C 1 3.1 142.8 146.6 110.8 131.7 141.8 131.6

A*. CQz

per tef 26.01 23.8 24.93 18,47 a. 95 : , 21.93

CO3 per

gat. day 6.5 .3 6.23 4.3 5.5 6.3 5.95

CO2 production in Ca(0r

Oa(0B)2 In

400 co. HO 10 15 20 18 30 35 0

3/15 25.3 .1 *?. 30. 33.3 36. 19.7

3/16 I.. 6 30.8 .4 , 19,1 30*0 17.6

3/17 33. b .1 .7 . lead 14.1

3/18 . .0 . .1

3/19 x . , ;. . „. . ,

Total C03 111.1 115.2 127.4 148.7 53.0 6b. 74.6
Ay.COj

per day 23. 33.04 . .74 26.5 . 14.92
COj per

graa day 5.55 5.62 6.37 6.91 6.73 7,13 3,34

i> of Hornal

CC 85.4 96.8 .3 161. 130.7 134. 68.

• later oont^&in it^i by Ba(OH) .

Ca(QH)3 Tad. Lenrth Length
in 400 no. in an. ro&ovtd
oo.Hw0 In no.

begun 3/19/18

TABL? 13 KXPFRIMTHT 6
Regeneration in Base* ■> Ca(0H)3

Length Regenerated in

3/23 3/35 3/37 3/30 4/3 4/5

4/9

4/13

4/17

4/30

10

1 K

10.5

1.

1.6

.

3,3

3.8

4.1 4,1

4.1

4,1

4.1

3 45

9.9

0.9

3.0

3,8

4,0

4,5

5.3 4.4

5.5

5.6

3 43

11.0

1.3

.

3.0

3,3

4,8

4,5 5.0

5.0

5.0

5,0

4 43

ICC

0.8

3,0

.

3,7

4,3

4.4 5.3

5,1

5.3

5.0

5 41
total

1.1

1.8
9TTT

lSfET

VK7

4,H

3*fir

S.Q

3TTT

10.

18.3

26.

*J4 ,t>

43.

45. 46,

4a,

48,

48.

15

1 43

10.5

i.i

1.9

.

3.6

1*4

4,8 4.9

4.9

4,8

5.0

3 43

11.0

l.i

3,0

3.0

4,4

5.0

5,0

.

4.7

3 4?

11.0

0.8

»C

3.0

3.8

4.4

4,8 t>» 5

4.7

.-

4,5

4 46

10.0

1.0

3.1

3.1

3.9

3.9

4,1 5,0

.

.

.

5 43

total

10.1

*

sSr

1.9

•

14Y»

lStS"

3T7T

4.5 3^8

37rjr3Trtr

4.<3

33TT

327"

t

9.5

18.9

38.

35.

41,5

44. 45.

46.

44,4

43.

30

1 48

10.0

0.7

3,0

, ?

3.3

4,0

4.7 <

4.5

ieai

3 41

1C.6

1.0

1.7

3,7

3.0

4.0

4,1 6,

.

5.0

3 44

10.0

1.0

1.8

3,7

3.6

4,4

3,8 4.1

4.3

4,3

.

4 41

.6

1.0

l.S

.

3.1

4,0

4.1 4.1

4.1

4,0

4.0

5 41
total

k-

1.8

3, 1

3,3

lTTS"

if!7"

3,7

~7T

4.3 .

..-It sTTo

3.9

•

ITT"

i37T

.

17,7

34,5

31.5

.

40,5 41,

- .

41,

41.

35

1 45

10.4

1.7

..

3.0

4.4

4.5 3.7

3,3

.

4,0

a 44

10.1

1.0

1.5

3,.:

3,3

3.0

4,1 4.1

4,1

3 43

10.0

L.

1,8

3.3

3.1

3,7

4,5 4.5

4.5

.

4,5

4 45

10,0

0.5

1.7

.

3.3

3,8

3,7 4.1

4*8

lea.i

5 46
total

XT"

1 . B

37?

-TT*

ITT

187T

■• ilk

"T~

lTT*

.

14.?:

.5

39,

36.

41. 41.

43.

4*.

43,

30

1 43

10.

1.3

3,0

3,0

.

3,7 3*8

.--

Aead

*

7,3

11.

18,3

15.

. ■.

. 36,

36.

0

1 45

10.0

1.0

1.7

3.7

3,8

4.1

4.3 4,4

5.0

.

.

3 49

11.1

,.

3,0

3,7

.

4,5

4,5 5.0

5,0

5.0

.

3 43

10.

.

' .

.

■...•

4,5 4.0

.

4,3

4.7

4 43
total

&T

1.9

lTT^

lflT"

•

4.4

"7T .

r.

4.0

4.4

4.4

10. s

18.

35.4

33,3

40,

41, b SO

33.6

33.4

31.8

44.5

44.

43.

fl.5

Approximate PH
initivl after 48 hrs.
9

10

.

7,

•

6.7

tablf 13 ^xp-himtht i

50

Refine ration
NaOH Length Len;;th Len
per in rns* removed
liter in mu

begun 4/16/18

in B'199* - NaOH

th in

H;-:en-r ftt#4

M

BG

18

bo

IT

3P
3?
37
35

37

•3.
38
36
30
tctal

33
40
35
41
If

38
M

35

total

38
39
38
3B
37

11.0

•>.
10. 3
10.3

4/ 0

1.0
1.0
0.8
1.0

1.0

10.1
10.4
10.4
11.0

0.3
0.7

.<

4/J3 4/33
1.6 3.7

Ti.O 3.0

3.0

2.0 3.0

1X75
37.7

.0

3.1

1.6
1,5

i.a

1.8

2.1

isfy

10.9

4/26
3.1

.
3.7
3.

'■' »

lETST
32.

Si

2.7

3.8

,.

i^TT

.34.

4.1
3.8

4,0

4/30
3.7
-.

4.4
3.9

4.0 4,1

36,
3.0

3.6
3.0

1 . •

33.
3.0
3.7
3.1

3.0

lSTF

30.

11.0 dead 4/19
10.0 dead 4/30
10.0 • •
10. S • ■
9,3 dead 4/19

no regeneration

18

11*0

40

10,

.

37

10.

.3

35

10 1

► ©

56

9,

Li.

tot

t

lb

11.

10.3
10.8

io. e

10 •

ll.o
10,1

0
35
50

1.0

.
1.0

1.0

*

i.<

..

0.7
0.7

•• .

3.1
3.0

1.8

xgtff

19.3

1.6
1.1
1.0
1,1
1.3

rrr

3.0

3.0

3,0
3,

.

3.4

3.3

lisir

1.8 1.3

1,8

1.9

|0

3.0
3.5
4,0
3.9
3,6

36.5

3,8
des4
.

.6
,6

4.0
4.0
3.9

!

deed 4/19

dead 4/30

• •

dead 4/19

* *

■</■■

4.1

4.8

4.

4.

~7~
43.

3.0
3.4
4,1
3.1

177"

33.6

4,8

4.3
4.4
4.2

'■< 1.5

3.0 3.0
3.6 3.0
3,1

~T 157T lOT iltT iTTT

5.64 11.9 17,3 19,6 26. 37, 39.3

no regeneration

Approximate PH of solutions
initial after 34 hours
7 6.5 to 6.6
8.9 to 9 8 to 8.3
10 or more 9 to 9.6(4/30)

tabis 14 Txvrmmm 8

Re^tneration in Aol.t - H3PO4

51

H3PO4

in 300

CO.H2O
35

lc

Length Length

in mb» removed

in mm.

50 13.1
53

4fl 13,5

.otal ^uiT

J

46 13
K 13 .9
39 13

47 3,1

>otal ~~T~

$

M 13

43

39 13,3

43 13

'OTAL 5STT

51 13

43 13.3
43

43

otal 1ZT7?

51 13.1

50 13.9

44 14

33 U

►otal 5X7?

53 13

ai «•*

45 1J.1
39

totU fStT

I«CHP04

13.8
1 .
13.4
1

srnr

Length Regenerated In
Begun 4/4/18
4/10 4/14 4/16

1.4

1.0
1.8

3.8
3.5

1575*

T7T

1.8

1.7

l.ci
-_

1.0

1.9

i.e

..

1 ..:■
~T7r

""XT
3.1
3.3

3.7

4.0
4.1

3.9

-TTT
1.9

.

l.a
O"

—7-
1,9

1.9

r • «
14.3

3.6

137F

3.3

3*1

i<-j#

;.

" H|' 111 "

3.0
■•* . ■

3.5

'■i 4 .
*TT

3.1
3,0

4.0
lTTT

..

3.6
3.4

.
XX

4.0

4.5

IS.

4.1

■ .'

31. S

TT

4.1
4,5

.

'■ -

«

3.5

•.

lTOT

33.

4/18
4.5

■ •
4.3

loTT

.6

4/31

4/34 4/30 4/30 5/4

-*77

4.0

•.

4.3

•

4.D

4.0
4.8

36.

—r~

4.3
4.5

4.8

:"T
35.4

T*"

177T

4.3
5.1
Ski

dead
t
«

47T"

TT" 4.0 4.0

4,0

.

...
iTTT

aJTT
4.9
4.7
5.3

HOT

tJt-

i.
..

1577

&

jl

4.n
4.9

...

_.

40.

S7CT
4,9

.

~T"

I .

"TTT

1*1

37.

™r ~

4.7

4.9

...

._

5.0
5.0

.

41.

~!*7r
■'■>. ■
5.8

COT

39.

— r~ -

4.9
5.0
6.1

43,

4TTT
.flfiL

...

♦7TT

3

I ,

■ ...

40.
5.0

5.0

5.0
6,0

- imi»

1,4 3,7 4,7

1,9 3.1 4,0 4.3
1.3 3.1 4.4

j.,0. 3.5 4.1 4.9

TTT 13TT 11CT lSTT

13.8 36. 30.5 35.

6.3
4.9
4.7

~7T

5.0

4.7

a. 7

«■* 1 . . 1

41.5

6.1
5.0
4.9

.
43.

5.1

3^.
45.

0.0
5.0

5.0

■■■.-

alTf

4a.

• .
5.1

If*"
45.

Approximate PH of Solutions
initial after 84 houra.
3.4 6.7

4.0

.

•:.■>

5.7

;.

6.3

.■

-.

•.

TABL5 14a EXPTOIMOT 8
Regeneration In B&aee - SaOH

52

KaOH Length

Length

Length Regenerated In w».

in *OC in mm.

remodel

Be^un 4/4/18

co.fUO

In hub*

4/10

4/14

4/16

4/18

4/31

4/34

4/36

4/30

5/4

0 51

13,1

1*3

3.1

3.9

4,9

'•

5.8

6.3 5.5

6,5

44

13

1.3

3.1

4.3

4.9

5.

5.6

6.0 8.1

8,4

49

15

1.9

3,3

4.3

5.0

5.0

5.

8.8 5.3

' » 'J

39
total

11.3

If

77T

3.4

4.0

4.5

~77

-.

S«3 5,-j

IS. 9

13.9

,.4

.e

' .

4 ••♦

47. 47.

47.

6 60

v$w

4.7

• ♦

«>.

1 B.B1

1 . ' ' .

.

48

13.4

1.3

3.1

•

5,0

k.

5,8

5.9 ■:. ,

5.

48

12

1.9

3.5

4.1

4,3

5.0

5,8

5.6

5.8

44

{J

73*

3.3

iI7»

4.0

iTTT

4,6

*~°

33,8 ; .

Ail

13.1

14,7

•♦,

l*i

38,6

.6

44.

■• »

10 50

"XT"

3,fi

3.8

t?i

■"XT"

b.I

!>, ' ... 1

CI

45

1,-3.9

1.3

3.4

3,6

4,0

4.0

4,5

5.0 5.0

5.0

45

IS.

1.3

«.d

4,0

3,9

4.1

4,5

4.5 4.9

4.9

3d
total

11

41?,

1.8

W7T

3.0

3.3
lT^T

4.3

lSfS*

4.6
IT??

5.0

i#7T

5.0 5.3 ^j^

.'■-.'■

13

.>.

30.

.

39.

41.5 4*.b

■..'../

.5 54

T."

.7

i3.«

3.1

"TITOh

47

13.4

1.8

3.9

3,6

4,0

4 .

4,3

4.8 b.e

5.0

43

.5

1.1

3.4

4,0

4,0

4.5

4,9

5.0 5,

I,

33
t*tal

13

oTTT

§ft

lO?

3.7

iSTT

ill lai.

■• « •

143

J

13,6

siii

;• •

.4

».

-.

. .

i 3.8*

15 M

™UT"

•.o

3.7

3.B"

.

Wp9

•

43

IS.

1.4

3.1

3.

3,9

4,3

4.5 4.0

4.0

44

11

1.1

.

3,1

3,5

I4

3, a

3.7 4.1

4.1

43
TOTAL

17.5 51

_—

1.7

ST?T

■ . •
iTTT

ifiT

3,9

iTTT

4.

1ST"

it£ 4.s
i~ iTTS*

4..<u

iTT

13,7

11.4

-. -

.4

,4

.4

.7

33. 35.

35,

■Jgf "

.1

i;4

:.

¥.o"

III

i.'r-Tnr

45

.5

1.8

.1

3.4

3,3

3.4

3,3

4.3 4.b

4,&

40

1 1

1.5

•

3,1

3,6

4.1

4.7

4.7 4,9

5.0

40
total

1 "% Sk

1.8

ST?

iTTT

3.7

itnr

4.0
1~

Aead
T7

13TT iTT"

14T"

13.5

.3

.• .

■ i.D

.4

30.4

• .

36,

f;.

SO 50

"■." '

•

w , 'V

1.4

■■s, s

3.7

I«0

4?

14

1.1

3.0

.

3.1

3,3

4.0

4,0 4,1

4.1

40

1 .

1,3

■v . 7

3.5

3.5

3.6

3.7 3.7

3.7

41
tot

■

US

iTTT

•

3.7

iTTT

lTTST

1ST*

4.3 4.5

lCT ITTT

4.0

iTTT

i

10.

•

**4,

30,4

•

30,

30, 3 .

31,

Approximate PH of Solution
initial after 34 houra,
7 G,7

7.1

fi

P. 4

1*8

7.5

5"

TABLE 15
Regeneration in Acid - HNO3

KX, NT 9

M»BKO~ Tad. Length length Length Regenerated In tm»

in 200' no. in mo. removed

cc.iuo in ma.

Date - egim 3/ 0/18 3/13 8/1* /::. 3/29 3/31 4/2

(>

Total

t

otal
7.5

Total

10

Total

12.5

Total
t

IB

"otal

1
2
3

4

1
2
3

4

1

2
3
4

1
2
3
I

1
2
3
4

1
2
3

#

40
30
44

14

43

43

38
30

34

30
41
45
40

43
39
36
38

47
40
38
37

10.4

II. 0

10.7

10.0

T7. 1

10.0

~4Tnr

10 .0

10.0

7.0.2
11.0

10.4
10.8
11.0

371 *5

41.9

10.0
10.0
10.2
10.8

41.0

1.0
1.0

1.1
t.o

1*0
1.1

t.o

1.1
t.

0.7

V

1.0

0.0

a. 8
l.i

l.t
1.0
0.7

0.8

3.0

f).*i
0.5
0.0
0.7

6.0

2.7 3.6 4.4 •!.

3,3 3.0 8,8 1.0 4.5
.9 killed bv aeold ■ nt.

3,1 4jl_ 4^ 4-fl 4^

12.0 15.3 ^J.l 107f 1'.

28.0 35.0 40,

3.4 4.1

8.9 3.8

'..:» 3.5

0.8

0.1 27.5

3,0

iTTf

2.9

2.1

iTT^

•1.

3.5

' i . 8

ioif

4.6

4.0

4. "J

itIi

1 1, 1

4.3

4

9.1 20.8 32.0 39.0

3.0 3.0

2. a

2 4

:.' ti*«

2.8

3.1

2,4

2.4.

10,7

25,5

2.0

1.4

1.9

2.1

T7T

' ,1

»> a

«J>. «-

0.0
3.0

lint

3.

3.0

3,0

2.9
13.0
81,
.3

? 1

dead

2.3(1

67T

4,1

9,1
4.1

1™
37. v

3.4
3.7

4 . l

3.4

1 CH

05.

• ' -

2.1

S.u

4,5 4,5

-1,8
4."

1.''-

lO
•a

f.
fc.i

4fl

I - ..-
40.

•1,5

1.1

4.1

4,

, 3
. I

a.s

3.8
.0

3, 8

37.
.3

dead

.'

4, 3

I . ■■

1.1
4.1
1.5
4.1

10. 8
40.0

1 . j

4.0
8,9
3.8

1 u . T
9M • I

4,0

3,5

urn '■■■

16il

dead

2.q)

5.0
5.0

n.o

4*8

4.8

iSTo"

46

40.5

4«8

4.5

^•0

5.0

4.8

4,9

^r

i^to*

44.

44.

4.1

4,3

4.4

4.7

4.5

4 .:■
1770"

iftf

42,

42.

4,4

4.6

4,3

4.3

4,5

4,5

4,0

4,0
1774*

40,

40.

4.0

3.9

3.5

3.6

5,0

3,7

11 .2

. B

- .r

5.1

5.0

a.o

I -.

3,6
ti

5.0

O.C

iO

4/4
6,1

5.0

5.0
15VT

4.1 4,1

4.4 4.2

4.7 4.3

4.5 4.7
\Wy 17TS

42. 42.

4,7 4.6

4.1 4.1
* .5 1*8

|.0 *s&

40. 40.

Approximate • of Solution

initial after 24 hro. after 88 hr«,
7 6.8

40.5 46.6 5,4

4.5 4.5

5.0 5.0

5.3 5.2

4.jp

iSto* lCST

45. -*5.

0.6

dead

dead

3.6

7.2
34.

4,5

3.8

3,4

3,2

8.<

.

9

.

5.4

6.1

4*9

4.4

..

80.0 -v.O

A*«d

17.5

Total

i

20

1

3

*

41

40
40
41

41

9.0
10.2
10.0
12.0

Irs

10.8

0.7
0.7
0.5
0.4

5.1

dead

1.9

1.2 dead

l.QCDl.o.

4TT

12.7

dead

8m4

1.1 dead (others died with-
out regeneration)

3U1

3.

4.1

0.4

9,0

5.0

v

TABLE 16

exp :t 10

negenoration in Acid - lMr.

cc. ad, Length Length
in 200 no. in hi* removed

Length Regenerated in

CO.IKgfl

in mim

Approximate

Rogun

3/12/18

•• — • ** •*

-.3/10

3/10

8/80

3<

3/31 4/8

4/0

of Solution

0

1

50

12.0

1.1

3.2

4.4

4.8

5.2

5,4 ' .a

B.3

2

47

12.5

in

-.7

3.8

},.

'.0

5,0

4.8

7

3

'•1

1.0

2.3

3.3

iOr

4.0

4-0 iifl

4.1

*otal

372

B 0m

11.5

lTTT

?TTT 1475"

tTTT

1

60

1 1.0

9,3

■ >

33,

37.

it,

42, 42

48.

9.9

0

l.fi

1 To

3.0

' 4.1

5.

~7T M

2

48

10.5

1.1

2.

3.8

'.'

'.

|08 4.2

4.2

5.5

Total

10,
."8

1.1

3 , &

2.5

.11.3

inTT

A. 9

,4

4,3 4.3

1,^,7

1

51

■ ;.0

10, <<

iZ_

32.4

37,

38.1!

.3 30.3

30,

7.5

1.1

:}.(>

1.1

~ rr«

5.1

5.1 |J

8.1

2

48

11,1

.1.0

2.2

3.5

1*0

4.1

1.2 4.2

4.2

4.0

Total

3

11.3

Sfs*

7?t7

itTS

iTTo

4,(5 .

ITT"

4.7 4. ft

I47?r lTTC

4.8

lTTf

1

84

13.0

0.3

23.

«j « . » > ;

30.

10.

41. 41.

41.

10

— ._

" Ko

ri.4

Kt '

4.6

4.6 •"T,?i

2

4G

U.6

1.0

2.1

3,5

4.4

tt

4.0 4.8

4.8

;:.0

Total

3

48

33777

178"

rrnr

3

— Ji .■«■

10.1

■ ■:.7'

ii7?r

'3.7
71 1

lift 4*£

3y7
77

I

12. 8

8*1

is. 7

30 .

30.4

1 ; 1 I . * .•

• .

3*;,

.

8.1

... ^. T ..

— •

2.0'

■ '-,i,jr' »

.

3.5 3.0

V

2

JO

11.3

,0.9

4»a

4.2 1.2

4.2

3.4

Total

3

44

11,0
35TT

H' ii"
...

CT

*» ,8

iT??r

lTT?

i17T iTTS

2*&

ii77f

1

11.0 "

7.1

18.2

it.4

■ »2 , "

- ^3.

33.

1

0.0

2 . J

" ^r"l9"■,""

4lT

'"3.0 :

dead

1

48

is. 3

0.7

2.?

3.Q

3.0

3.0

dead

.3

Total

a

1 1

H3r

....

7\S

TTTT

3.1

r».'

dead __.

— Co

91

, 7,«1

1Q.0

22.0

,

M58.

•i..

i?.r>

i

47

18*0

1.6(7)

nau

2

4i

11.8

dead

3,2

■

45

10 .1

w

Total

J^JP

<

.3

20 All died without rogoneration.

3.1

nepene

06.Ho ,i
in 200
OO.HgO

:>ate -
0

ui
5

Total
t

r.r»

Tota:

i

10

"ota!
12. B

Tota
18

Tota:
17. fi

Total
20

otal
(T) il

55

rABLK 17 NnUIOMf 11

Regeneration in Acids - H2S04

oo. Ho ,04 Tad •Length Length
in 200 no. in erurerooved
oe.H^1' in m.

Hate - ne«un 3/0/18 — - -

! cnjrth Regenerated in vm.

tal
S

Total

f

7.5

Total

"otal
12.5

Total
10

Total
17.5

Total
20

1
2
3

1

2

a

1

1
2
3
4

1
2
3
4

1
2
3
4

1

2
3

4

1
2
3

4

1
e
a
•1

41
43
40
SO

10

43

40

36
20

30
0S8

43
30
37
38

4S

43
37
36

40
41

•11

80

30
43

56
3e>

40
08
36
35

11.0
10.7

I- .1

10.0

10.1

lt.0

11.0

TZTf

10.0

11.0

0.1

10.1

10.1

10.0
10.2
11.0

VC7T

10.5"

10,0

10.5

10, .3

10.0

10.0

11.0

10. a

4THT

11.0
10.4
10.6
10.1

otal
(T) Indicates

10.4
11. 0
10. 0
10.1
TT3

- 3/13

l.o
t.O
1.0

TH7

0. 1

— Y' >■ -

.5/17

3.0

3.5

3.4
3.3

i?Or

31.0

3.6

0.8 3.0

1.0 2.0

3,0

1TF7FT

T..i :8.(,

-rti — TX

l.l
1.0

•*0

v"

2.0
2.1
0.1 2,2

?~T Tot?

0.0 25,f3

ole 3.6"
0.8 2.5
0.0 2.3

37T To7,i

8.2 24,2

*<~7o* 27T"

1.2 2.6
1.0 2.8

3/20

4.0 5,0

4.3

4.0 4.7

''"IT 1.' "

! ■

4.5

.13.

.11

4 .b
4.1

o.r>

■47. *

3.8

-.

3/,>r»
5.0
1.1

'.

ltCT

15,0

4.5

3/27
#,8
5.1
5.0

a

3.8
3,0

ITPT isTTT
34.0 37.

~XZ — ttt

-.'■■
. **
2

03.0

0.8

—

2.0
Co"

23.5 .

170 57FT

0.8 1.7
0.8 2.3

3.0
3.4

itS

20,0

t.i

3,3

iSfo"

99

1 ...—-■. .. ...

4.0
4.8

4,0

1673

37,7

1.3

.
3.1

570

4.7

3.5
3.0

ia77

40.

~17r

:.

tut?
30, >•

45. ,

4.4

4.8

44.

■-"sir
4.3
3.7
3 . *

ioT?
*2j&

4.F
5.3
5.3
4,8

27Ot

48.

3/31

4.8

5.0

4.8

!$7?"

47.

4/2
5.0

5.0

5.2

25T2"
48.

478
4.0

a,g

10^7

4(3.

0,0

TFnr
8i3

T7T
0,5
1,0

ii . <•

0.3
0.5

gjtf

6.0

0

21^0

1.0
1.2
1.7

.13.3
dead.'
1.3(1

1.5

dead
27ff

13.0

3.0 .7

3.0 O.

2.2 2. a

iTT3* lirnt

27,0 32.3
dead

1.3 1.3
1.7(7)2.

1 .1 dead

i.o

4,0

13,6

Si .

3.8

lTTT

4.3

3.8

4,0

3.5

uo

iSTS

38.

3.5
3.6

%o
33.

?cr
4.0
4.0
3,6

ifTfi"
43,6

4.3

4.1

177T

.4 2..

4.1

3,7
IJ

1674"
40,

3,6

3.6
UP

1377*
33.

4,8
5,0

48,

4,0

4.0

3.0

lTTo"

43.0

4.4
4.1

111
If^f

42.
*^75"
4,2
1. I

tffS

.40,5

^_

3.4
3.6

33.

^r.7

5.0
5.2
5.3

2(177
4' "

4.*

4.0

3.(3

1775"

.0

S.fl

4.4
4.1
4 . 4

17VT

4.0
4.0
US

itTo"

4.0.5

—>r?r

dead
3.8

3.0

iottf

Approximate P3 of solution,
initial after artcr
24 hrs. Jirs.
7 6.8 C.8

5,5 6,4 6.5

4.5 6.0 6.3

4/6
4.0
5.0

o.3
4.0

7?

5TT

5.1
4.7

5,3

2TCT
48..

3.0
4.0
3.5

177TT

43.0

•nrar

4.4

4.1

4.1

177?

"^tr s.4 4.0
4.3

4,0

4,5

.IT
40.5
•U'^d) no furtlier growt

3.3 4.
dead 9 3
dead
377*

3.0 5.5

' .

■ ■ — »'lW ■»!

1 , ;i dead

2.) 3.0 3.1 dead

173*

3,2 not determined

)1.3

1.7 2.0

1.8 2.1 2.1

Q '1

- • ■ •

3.1

3.

tadpole was transferred to neutral water

ec.l

cc.I

Date

0

Total
%

Tota]

f.fl

Total

10

Total

i

12.5

Total

15

Total

TABU 18 13

56

Regeneration In Acids - 1-3^(14

00.1131*04 Tad. Length Length Length Regenerated in vn.
jn 200 no. In nsn.retnoTed
cc.!I2<> *n tm»

- ttogun 3/0/18 43/^ 3/17 3/20 3/23 3/ ^ 3/31 1//Z

4/0

Approximate Pli of :>olwt
Initial after after

24 hrs, hv .

0

1

10.0

1.0

3.3

3.0

.

4.

4.6

4.S
6.5

4.

2

40

10.

1.0

3.4

.

5. 1

1 .

n

43

10.7

1.0

3.5

4.7

4.

5.|

1 t

207?

48.

10"j7?

5.0

20.
48.

0.
49.

Total

41

11.0
1 .6

r
0.

31.

4.0

15.~
37.

r

i' .3

1 .

5

1

•14

11.0

1.0

3.0

3,0

3.8

.0

4.7

.

M

<.•<

4.8

Total

•

3
4

32
30
40'

10.0
10.6
10,

12.5

1.0

i.i

0.0

3.0
28.2

4.

15.3

>.

1.1

17.
42.

1.3
4.7

4.7

4.8

5.7

lTTT?

47.

5.0
47.

1.0

27777

17.

5.0
4.8

..'
7"
4h.

0.0
4.*>

i.o

7T

7.0
Total

1
2

3

4

44

30

10.

10.6

'>.0
1 .7

4THY

0.0
t.o

.
1.1

r
1 ». •

.3

.8

.(>

3.0

10.0

27.0

1T.3
35.3

4.

1.1
4.

1.

43.

4.2
4.5

4.3

44.5

5.0
1^72?

.•
5.2

47.

3.0

4.

0.0
0.

47.

10

1
2
3

45
38

.0
10.0

11.

0.8
1.1
1.0

1.0

.0

3.5

4.7
3.

•1.

5.0
L.I

40.

0.6
3.7

1.

i|

' .•'
41.6

6.5

4.

41 .

6.0
4.0
5.0

'.'

io7T

44.

o.s

4.0
0.

44.

Total
*

4

37

1.0

3.'

8.0

2.7

lTTfiT

.5

jO

12.5
Total

1
2
3

4

40
38

10.5

1 .
10.4

11. >

1.1
l.o
0.
1.0

0

.0
. 1

.7

.

1 1.
34.

4.

3S.5

1.

4.6

4.

1.1
17.
41.5

«.«
■'.<
1.4

1777*

41.5
5.0

5.1

4.8
4.4

lTf7
U.5

4.H

8.1
• .8
4.4

3.4
1777"
41.0

4.

6.1
4.8
4.

lTTT

41.0
4.7

4.8
4.4

41.0
4.6

15

1

41

10.0

0.9

.

4.4

5.6

5.5

0.4

5.4

1

11.

.

-.7

.

.

.

4.6

4.

4.9

0.0

3

4

ID.
11.0

1.1

1. 1

4.1

1 .

r . >

lfltff

.

0.0

t.o

•joTT

Total

nnrr

~

lTTa

15.

.

.

44.9

47.

17.

47.

47.

9.5

.

' .6

.7

I .:

.

6.3

.

. ♦

,

J

0.1

.

4.7

>.'

.

Regen

CC.li-i

In «0i
co.wa

D ate

17,5

Total

20

Total

22.5

Total
t

10

Total
t

57

TABLE 18

EXPERIMENT 13 Continued.

Regeneration in Acids - H3PO4

cc.il <jP04 Tad. Length Length Length Ttegenerated in rsm»
in 100 no. in rw. removed
ce. water* in rom.

D ate - Hcfiun 8/9/18 3/13 3/17 '\/~) 3/37 3/20 3/31 4/2 4/6

*7.5

rotal

20

Total

1

2

a

4

2
3

4

30
35

42

13

37
30

10.0
10.2
10.0
10.2

10*5

10.0

10.0

10.0

1.0

V

3,8

0.5

2.

3.3

1.0

2.0

3. 1

1.0

3T5

lis

lTT?

4.0

iTHT

8.0

- / . M

35.

0.7

*• • ■

*«*

0,6

0.0

1.0
8.5

2.8
2.8
3,<)

3.3

2.7

JO

•1 , 0

I,

5.0

10,3

4. J

o7

3.7
4.4
4.5

4,8

17

4.0
4.4
4.5

3.5

3.5

* •
3.S

3,5
3.2
4.1

'"J.n 4.0 4.3 4 ,0
33.4 37. .7 3*»

3.8
3.8
4,2
4.0

15.*,
38.

•1.0
1.1
4.2
4.4
1ft. 7

I .f, 41.0 42. 43. 11,

3.8

4.0
4.2
4.0

ic7o

38.5

4*0
4.1
4*0

43.

4.0
4.1

4 .3
10. b
41.

3.8 3.8
4.0 4.0

4.2 4.2

4.

4.0

1M -

3tJ.fl 3b. 5

Approxinate I Ml or solution
initial after after
hrs. hr«.

.2 "i. !

5.5

3.0 4.8

.

22.5

Total

Total

1
2
3
4

1
2
3
4

47
41
40
40

40
43

41

38

10.5

l.o

2.9

4.0

Arad

10.4

0.0

b«o;

3.0

.7

4.0 4.0

11.2

0.0

3*. 8

3,0

3,3 3.2

10. 0

TTTT

1.0

2.4

iirnr

1~?7T

dead

~7r

rrrr 772"

o.

25,

33.

.

i, 33.

11.0

0.6

2,3

0 -.

3.4 3.2

1 ;.0

1.0

2.0

<•*

if .A

dead

10.0

0.0

2.4

2.8

.id

10.0

TT.IT

1.0

3.0

iTCT

13V?

Id 2t&

8.5

25.

. .

32. 32.

3.S

14*

3t5
(Of
32.

4.0
3.3

34.

3.0

30 1

4.0
3.3

773
34.

» -i . B 3 . !

8*0

30.

4.0
3.3

773*
34.

3.2

3.0

G.O

SO.II

3.6 1.7

3.4 4.4

4.0

tahij: 19

IT 13

A. Normal C02 production

gttS.

No. Wt.of u<io produced

Tads,
date -3/2C 3/30 3/31

Total G02 ]>«**
OOs day

1

2.'

m.

ii. i

10.8

an.!

in. on

•-•

0.0

17.0

16,

17..1

15. fe

n

.0

•6

.

1 .8

1 7 . 0

15.8

1

2.'

15.7

14.0

Ki.l

45.6

15.26

a

J. 6

ia.7

11.

14, 8

40.4

13,

ri

J. 7

16 .a

13, >

ia.a

42,0

1 4.0

7

o.o

14,

11.

io,n

40.1

10.4

6

o.o

i a .

IS. 8

IS. 2

46 mf

is. ao

0

*» .o

12, .i

10.

la

. 1

13*18

0

,6

m.t

,0

in. a

37.

12.43

• ay

fc.45

i -

B. COo production of the sa^e tadpoles In Acids - no] ad tlftr

i. cc.Hwl in
40© cc.ttoO

A*

date

06*

4/1

pr <.'duoc
4/2

>d Total

vsr

dav

sr

of inonnal

1

0

U.2

8.7

20.9

10.

3,6

.

2

10

o .a

7.7

17.0

• -

.

,8

0

20

0.1

7.

16,3

H.ia

■ .

.*

i

25

,0

6,1

14.0

7.0

,

.

5

00

cc.Mfcr In
400 cc.

0.0

1.3

14.3

7.15

8

10

6.4

10.0

.

*

.

7

20

7. <

3,6

11.0

.

1,1

8

25

7.1

a.^

10.2

.1

1.7

.4

o

00

6.1

.7

.8

1.4

1.

oc.

10

0

11.

0.

.7

10.05

4.14

.

Riren as 0.01 N U2CO3

30

13

-,.

^; oration in Acids - liCl and R1 r.

Vo. oc,li«Jl T.enjrth Length Tent!i regenerate;! in »n,
in 400 la r.r.ror.oved
cc.i'oii In fete.

date - Bestm 3/31/18—4/8 4> 4/0 4/13 1/17 4 #21 4/25

1

•

0

18

.4

1.

.2

3.0

.0

4.7

5.0

►0

11.0

1.0

.

2.8

3,4

.

4.'?

4.

42

11.8

l.a

,0

3.0

' * . &•

4.2

«.

Total

40

1 ,0

4T75"

. o

<

oTiT

3,1.

3,0

4.

1 7 . 1

4.*J

1 - . ">

4.4

17,5

*

11.3

If) R

2C .8

• -.•■'•

.

17.

.

2

12.1

8.0

3.1

»»•<•■:

44

12.0

0.

1.

3.1

4.

■

.

4.5

44

12,

1.1

•a

3.0

3.7

4.

4.0

Total

41

11.
4t7T

1,0

3.8

2.1

7*nr

2,8

3.3

iTTt

-

4tS

77

177?r

t

20

40

fcl ,.

16,3

iJi>. 5

31.0

38,

n

ii.o

1.0

3,0

4.0

4.8

■. .

5.0

12.1

0.0

1.4

2.1

' • • •

3,1

3.0

10.0

0.5

1,8

8*9

2.

3.

3.0

Total

11,7

27?

2.8

ioTF

I?T

3.0

'.1

1 B , g

3,0

' .ft

88

died

■ML

14,7

23

88,

33.8

4

without

regeneration

ft

30

•

i

«

fiiir.

10

Total
7 20

8

Total

Total

0 30

10 0

Total

50
47
1$
39

52
48

42
12

48

10

died

8ft

41

42

12.5 1.0

82.3 1.4

11.4 1.4

10.2 U

TE7T T?F

10,8 37.7

i.a

2.0

|

5 1

■ - . -.

8.2

12.5
12.0
11.2
11.8

'""7"

i.i

■
*• . ■ *

3.0

2.0

iO

23.3

O.G

.
2,1
1.5
2^4
8.6"
10.!;

3.0
2.fc
3.1

127F

...

2 died without regenerati
.3 0.3 0.5 dead

12.1 0.0

mrr itr

3.1
3.«

3.1

;:.n

20.

11.

i.

3.6
4.0
3.0

141F8

31.5

t.o

34.

4.0

•1 a

a

3.5

i17??

31,0

(.;

4.0
4.0

',1

on

1,5

0.8

dead

0.5
4.0

jjtii :*a". .

ltliout regeneration

12.5
12.4

fen>
7f

1.3
1.3
1.8

11.1

18.1

3.0

3.0

lTTF

3.8

i
4.1

4'J i.V

5.0

4.7 4,7

"'77

r.3 37.4

itT^

60

T 31 * 14

\. Xorral Carbon dtoxid product i on in distill' ter.

Date Deo. 17. Dec.l . DM,10, DSO.81. :jcc,21. Total

Tadpole OOfl produced as 0.01N iij>i>o

I, 0.4 7.4 7. O.fl * ' .

5.5 .1 . 5.5

8. 6.1 7.3 ($.85 0*4

.7 .7 7. .0 .5

B.Carbon dioxid production in aciu - DC1.

Tadpole 1
n/i 1

in 200 c<
of water
Pfl 7

10

4
tr> Tadpole 1

3

3.8 3.2

ate COo as 0.01N KgOOg.

V » ■

oeo.22 5.*>
,;

7.t
25 & 2611.
4.
?!«

7.0

4.55
7.4

s.o

5.3
5,3

.
5.8
6.1
6.0
6.0
6.5

,

6.1

<>.7

.
7.5

2*>

80

31

Jan. I

3

4

7

•10

11
I

1",
1 I

1G

17

10.6

V>.8

20

,7.5

22&2$

;

24

25

,

27

.

28

0.0

t.

4.3

10.1
4.8

7.0
G.3
4.4

.

. I
4.7
3.6

5.0

•

5.2

• > . <-

.
5.0

1.0
5.1

n.<*

..

M

.7

5.8

•s>.4

.

3.8

5.0

3.7

B . 1 0
3.7
3.05
1.7
2.5
I. I

2.0
1.4

9 '1

• «

1.7

'.

1.7
4.0
i~.l
,0
4.8
' .7

7,0

4.0

8:8

0.0
3.0
5.6
3.7
3.8

b >J an •
8,0 30^01 12.5 . .7

1.3 Feb.
1*8

■ • -

MM

8

7

10.0
11.0
6.3

9
10
11
12
13
14

1,4

6."

8.1

6.3
6.6

8.3

1.6

•.

3.0

1.7

1.8

1.3

0.0

I

1%0 SuMMary

3.3 Tir^
J *7^n days

0.0 'otal
1.8 M2
3.1

. r*c02

>er day C.3

3.5 C02 91'3

3.0

1.5
1.5

8.2

8.0
4.8

Par . 4J

3.9

.

6.2

1.6

2.5

ft. 6
dead

'.-

u

'.■•■
dead

1 . 3
teftd

333. 0 264.1
4.

* Experiment raised from 13 tb 20 degrees C.

G1

taw: ; 23

14

Regeneration in Acid - IICl lV#>> ''

Tadpole 1

T enjrth. Bny„ 70

2
71

3

7 ."•

Length
or tail

40

47

48

Lenfctl*
removed

13.

12.

.1

M/100 UC1
in 200 cc
water

0.

0

10

t?

Date

Amount regenerated

Jan.fi

lonpth
in nan.
1.0

0*

ler

In raw.

.7

length
in
.5

1 B

let:

1 n

^

13

1.7

12.

1.

10.4

.8

,.

.

.

20

£.♦

If). 7

1.0

15.

1.8

15.

0.

1.1

25

30

3.0

3.5

83,

.3

2.0

3.

23
3.6

10.

21.8

'.}.

0.6

Feb. 3

4.0

;kj.

:*..2

.3

2.0

21.

^.

i.3

7

.5

1,

■ » •

.0

,;.o

dead

12

11

4.5
.5

1.

34.

Head

.5

3«

dead

1

/>?

62

TABL^ 23 FXPFPI^TJT 15
ttel'ition of 11*4 ;i,n ! Gs-rVm Uttl I ProJu.ni .n

T&ipol*

i

a

It.ijaa.

1.6

0.86

D*t«

C0a

CO3 per

gVM

co^

CO3 per

9/25/17

3.

1.47

1.05

1.33

9/ae/i?

1.7

1.06

1.

1.23

9/37/1?

!■ -1 - II

1.44

1.3

1.55

TOt<|l

•#aa

3.97

3.4

.39

G3

TABL* 34 FXP*-RIMFNT 16
Relation of Sine and Carbon dloxid Production.

Ho. Tad.

Lta

;th

in am.

Width

Width Depth Area

Weigl

no.

total

of 1

-nil

of body body

tail computed

1 1

7.

43

15

11 11.5 1?"6

3.7

3

7i

50

)

15

11 11. 70

3.6

3

77

;>

16,

12 11 1318

1.7

3 1

H

39

33

TOTAL
13.3

9 1C. 77

a

a

33

13

10 10.3 1339.7

.

3

66

43

13.3

10.1 1434.5

win.

3.0

TOTAL

rrr

3 1

56

37

19

10

7 7.7 883.3

1.2

3

53

33

11.6

8 7.7 874

1.3

3

41

31

16

V.J

8.. 7.0 7-*4,7

J'l."

l.C

«T>rvr*T

*T~*

Carbon

Hoxid Production in Distilled later.*

lo.

1

3

3

Data

1/32/18

54,

1

4C.1

19.8

1/

46/3

3C

13.9

1/

pQ •

8

36.7

16.5

1/.

71.

45.1

16.

54

30.5

i/at

7.

31.1

.

Tot*l

^

lETTTr

CO3 par

lH.y

e>^ .

73

.

17.51

par day

g*aa

4.

4.66

5.0

Relation of Araa.Waifjht and CO^ production.
Ho.l tAken a© 100.
Araa 100 74,1 44.6

Hi :ht 100 64.17 .17

COi produced 100 . 32.

CO^ given aa 00. 0.0 IK H3C03

64

TABLF 35 FXPSRI M^KT 17
Relation of Siae and Carbon iioxil Production.

No. Tad,

» Length in

; :,. .

Width

Width

Depth Area**-"1 3 i ->>t

no,

, total

of tail

of boly body

of t aj

LI computed ,gas.

1 1

73

44

33

14.

13.5

lv

4.1

a

71

44

37

lo.

U.

13 1566

3.5

3

70

43

37

TOTAl

14

1

i

11 17

3.6

lTTT*

1

3d

38

.1

8

B 1034

.0

I

54

1

13

.

9 1041

1.7

3

•

4

11

6

7 984

S7T

TOTAL

^

3 1

49

31

18

9

7.5

6,5 768

1.3

3

34

TOTAL

9

8

8.5 83)

1.0

Carbon iioxil Production

in Oi^tillei Walter

Ho.

1

3

3

Data

8/4/X3

43.1

36.3

13.7

.

38.5

.

3/8

.6

.

9.3

2/7

55.

29.0

13.1

/•

38.5

15.3

5.5

3/9

31.7

19.0

8.7

• 0

31.9

15.5

5.4

3/11
Total

CO,

0.4"

323T"

16.7

7.4

CO3 par day

40«

21.

9.

CO2 par

graa day

3. CI

4.16

4.5

20 oe.HCl

per 400oo. Wa.ter or 0.005K jFfCl

•/]

.

.0

3.7

13

27.0

1C.

6.0

14

.

7.5

3.

15

32.0

13.3

2/l

.6

3/17

gjSjU dead)

T Total

C0.„

l^tTT

1 ', . -

1375

CO^ pe-

:y

1,8

11.

£'8

C03 pe

S».day

•38

3.16

Deo re** e from

normal COj, per

da. 15. 3

.3

4.5

Ratio of ft. Area aril Deore&ae in

C9^ takin f.l aa

100

;ht

100

45.5

17.86

Area

100

53.

30.5

CO^ Jeoreaee

L *

100

66.6

30.

65

'm-m

TABL^ 36 inrpFRIMTHT 18

Effect of Sl*e on CO-, production anl Regeneration
9fl. mm

Ko. Ko.of Av. Total Total

COn Produced

Tadet

length w% •

ur« t

3/9 3/10 1/11 3/13 '13

3/14

1A

1

79 3.6

181

81

84,

,3 19*8 lfl.4 81.

-

IB

8

64 3.4

1001

34

.9 33,

,6 16,3 16.3 lii.9

13.4

1C

6

44 3.4

3944

M

.7 33,

,1 17.1 17.

19.1

M

1

3.

3104

,3 30.3 30.5 18,7 17.0

18 -

-B

3

36 .

,7 30.'

7 18.5 17.0 16.3

1

20

7

40 .4

4164

,6

»•

J 19.3 13.7 19.6

18. 0

3A

1

75 3.7

iaa

8$,

.3 35,

,3 19.0 19.9 30.3

14.5

3B

3

n .7

3369

18

.3 19,

,1 16. C 15.9 15.7

18.3

3C

6

89

3547

16

.1

,0 15.4 13,5 14.7

14.7

ii

1

76 .7

1800

13,

. - 17,

»5 16,1 14.5 14.

11,

«i

a

.

1313

17

,6 14,

,7 12.1 11.3 1*.0

.7

4C

6

39 3.7

^i *i -"i> •.

.

14. 9 14.0 14.7

• 5

The aftite

tadpole* with 300,

,o. 0. .1 B KQH, O*(0R)a n,i Hr<S04

made up tp 400 oo .

summary

3/15

3/16 3/17

3/ J.

003 per

CO3 per day % of normal

Control

„y

normal

In expt. 00

1A

,

10.5 13.6

,

."57

58,4

IB

IS,

.7 14,

.7

.4

14,3 ft«

1C

1 .

.3 14.3

.3

30.

14.65 7 .

■l

M

23,5

19.0 19.0

30.8

17.37

15. 88,

*>B

24,9

23.3 SI. 4

31.6

18.8

3... 107.

*b.

dea*(5)

31.06

35.6 1,

0*445) <*

U

il.C

19.0 16.1

13.

8 .93

17.55

3D

36.8

31.6 17.5

15.

17.4

30.4 117,

50

24.3

3 dead

IS. 23

34.8

%M<

4*

1C.

7.4 ia.8

5.0

15.5

8.67 56.5

4B

1 . .

9.1

-

13.

10.3 78.

4C

11.

5.6

9.7

16.

8. 56.

8UMMARY Continual

Bo.

C0a ]

mi lay per 1

jfUft

Bo, CO a per grais per lay

normal In expt.

normal In expt.

u

»S8 3.56

31

7.7

xL

.7

4.

n

6,45

10

6.0

4.3

30

5.65

84

4.

4A

5.65 3.38

n

5.

7.

4B

6.0 4.7

BO

6.3

7.

4C

5.9 3.

06

TABLE 37 TXPfHIiPTrT'Oontinua 1

Effect of Si so upon COo Production *o& Regeneration

The Surririn ^oiee of the previous experlnent were operated
on<i aioj*o i to unler.TO regeneration in the eoae solution* ae
thoae in *hich the CO3 production had been determined*

Ko. So .of Length Length Regenerated in mi*

T ■....-. ■ {

Confrole
11 1

i

3/18/18
16

3/33

1.1

.9

3/.-1B

.1

13.1

3/30

4.4

-

4/3

4.5

4/9

.0
37.

4/13

6.
41.

4/18

6.
41.

18

a

-.

4.9

30.

7.4

30.

8.4

8.7

9.7
40.

9.7

40.

10

6
i

- . •

6.6
11.

14.7
23.5

.1

0

26,0
41,

39.6

47,

.

.7

48,

1 98
21

lt

.1

1.0
6.

2.0
.4

34.

6.a

6.9

7.0
43.

n

%

23.9

1.5

6.3

16.3

7.0

6.1

34,

10. 0

.1

.1
42,

4
P

■ •

1.9

3.7

6.6
Itt

39,6

6,4

6.5
31,6

Ca(0H)u
31 1

16.

1.1
,8

3.0

5.0
30.

6.1
36.8

7.0

4::,

7.0

7.

43.

3B

8

24.0

let

6.9

.5

.8

33,

10 «

43,

10. .

T'.8

3C

4
*

40.1

11.

.0

35.

14.4

15,1

,1

1

16,1

16.2

1.0
6.

.7

i.T

5,0
31.

43.

7,0
43.

7.0

7.
43.

4B

I
*

.

2.
9.2

4.3

Id.

6.3

8,0
33,4

7,8

*

7,8

.5

4C

6

59.7

11.4

.4

1,
32.

,4

33.

•
.4

deart

CT
TABLF 38 FXP^ItfTO 19

Sfftot of 31 se on Regeneration In Aoide - HC1

OO.HCl

".. .

of L*n"

th Lenrth Len

gth Regenerated in

•

p*t 1.

T-% l»

rOAOTOd

4/7/16

4/15

4/17

4/19

4/33

4/34

4/37

0

3

.

9.

10.7

14,0

15.

16.

18.0

$

13,

39.

.

33.

• 5

.

43.

3

43

36.4

5.9

7.9

.

1^.3

18,

14.4

15.0

%

16.4

21.7

33.

,

.

41.

3

38

16.4

.

4.8

6.1

6.7

7.5

8.1

8.. 1

i

33.

.

41.

• ,

49.4

49,4

0% .

■

51

43.

8,

9.3

ia.

14,1

15.

1

.■

t

16.4

23.5

. 7

>-.

36,

.

• 6

3

41

H.l

6.

.».'..

11.7

13.8

14,8

.

16.0

18.

.

34.

37.

43,5

.

*7.

4

40.5

9.1

,

13,8

16.0

.

17.9

17.8

$

- '. «

SO,

.

39.

41.

.

44.

Average

•

£ of Regeneration of the

two above Control*

6

15.1

„

•

,

.

.

40.3

6

43

17,

34,05

9

.55 39,75

,

44.15

6

30

33.7

.7

35.5

40,35

43,5

46.7

46,7

30

3

55

45.

5.9

9.0

.3

13,4

1:5,4

14.5

15.3

i

13.

19.

. V

.

. .'.

33.6

■a

41

33.4

5.8

9.

10.5

11.8

13,0

14.2

14.3

I

17.3

.4

31,4

3l>.

.

• 4

• 5

4

'53

,

.

10. 1

13.. 8

14,1

14.8

,

16,

r*

19- !T

a i» <*-•

»o. t»

•3 <* ■ to

37. *-

H).*

H-a-a

40

3

1,

4.0

• 3

7.6

.

9.6

•

1 /*

i

1.';.

ia.

.

3.; .

33.

.

,

3

43

3* .

6.4

9.0

10,

.4

13,

,

14.1

*

17.

I,

.4

34,

»■'•

•

•

4

33

37.7

7. <

11.6

1 ,7

13.

15.0

15.1

15.3

*

19.4

31.

.

.

40.

40.4

N

3

•6

44 .3

.

10.3

11.9

.7

.7

13.8

i

1.

19.

•0

37.0

39,0

31,0

31.3

§

41

'■■ «

...

7.

.

10.

11.8

11.7 /dead

7*

4

3*

s*o

17.3

,.:,

13. H

-

-

S3,

^

X.7

■3 3- *~

3*

5fir. 1

5 9

% of Regenervti
Control

on Conpejri?*
» • 100*

to the

Go^trole

30

81

,

e.

83,

.

.

a;i.

41

IOC

118.

102.

.

98.

99.6 98.5

M

86*

69.

• 4

90.

.

.

.5

40

51

91.5

.

101.

93.

.

88.

87.

48

t.

104,

96.

' .

91.

89.

88.5

33

.

104.

,

87.

93,

.

88.5

m

88

91.

84.

.

.

.

,

77.5

41

.

.

a:.

.

.

,

.

32

.

91.

...

.

.

e:>.

68

TABLF 39 TX^^RIW^KT 30
Regeneration in low (h.y

Jar 1 Z

co, ft)

per j.Uer

•tufcimuB 0 1.04

■lnUua 0 C.l

Average 0 0.54

Tadpole

length 77 73

removed 11. .7

l«£*!ier%ted

a oegu* 13/18/17

U/tt deed 0.6

1/6/18

1/1C

1/lft

1/30

1/
l/£3

i

:

i.e>
11,

•o

14.6

.1

15.

18,

-

£.5
1.0
1.6

73

1.0
B.4

••1

17.5

e '

ia.o

3. 5

31 ,

94.

35.

-.

§•0

£.48

77

1.0
ft.

1.4

11.8

16.C
17,8

■
9 . ;>

» -

ft. 04

7C

?.«

17.0
3,4

3.8
- •' «

3.4

3,0

ft

4.35

»

.0
16,4

.4

. -i.

16.

81,

.
4.1

4.7

70

1.3
10.0

I .

3.5

3.0

£8.

3.3
16,

3.
3.

■

CO

TABL* 30 FX^^riyFNT 30
Re gen* ration in Low Oxygen

Jar

co,0

per liter

6

wuil*u*

0

1.04

.5

.

3.

4.35

5.33

Klftlaijcp

0

D,U3

l«

3.0

2,54

.

4.1

f.verig«

0

0.54

1.

.

3.03

3.

4.7

Tadpole

) en^th

M

63

53

.38

59

18

50

removed

• 1

10.

10

10

.

Re. »nexuted

fcen*n

13/18/17

0.5

dead

1.4

c.

0.4

0.9

1.6

i

5.

19.

5.

4.

9.

.3

1/6

1.

3.8

1.0

.1

1,3

.7

$

1&»

2G.

16,

ii«

13.7

38,

1/iO

3,&

3.1

1,

•*r^

1.4

3.0

i

.10.

7

ifc.

14.

30.

i/ia

.1

3.S

1.9

1.

.1

i

u,

I . . 6

19 .

lb-

M,

i/ao

»5

3. 3

3.1

.€>

.1

i

25.

39.?

9*«

U,

.

3

.

.5

dead

.

3,

i

■v5.

33.

83.

33.

1/ »*8

2.5

3. 5 (deal)

3.3

9i

i

18,

33.

33.

Taipol

..

length

37

35

34

36

>**

36

34

removed

8

7.5

7

7.

7.5

8

8.1

Regenerated

i

deal

C.S
6.6

G.8
14.

dead

dead

(&Mfc 1

.0

1/6

i

%%%i

de

3.1

38.

1/10

l

3.3
41.

1/15

^

3.6
44.

l/~>0

i

.8
47.

1/33

i

3.8
47.

70

TABL? 31 fXPSallim 31
Rtgtneration in Low Oxygen Water.

Jar no.
CO.O3 p«r

liter

, 1
r.ini-u;:

averse

1

0
0*

Length

NMff 5.

in an.

17

17.4

3

>e 1.14
0.37

0.7

3 4

♦77 1.1
1.9

1

3.6
3.4
.17

Hit $
. 30.

7

4.68 5.0

3.3

3,68 4.1

Jar. Len

in an,

1

67

Length He

:.v'r

1.6 •.

.BTt

generated in
/a/17

2
am.

• 8 16
3.0 17.

k/3

• •

• -.■■..

3 71
65
A*.%

.4

17

2.0 ii.B
I87T

• 3'3 „

4, ->3.5

@. 4 3w.

6. 3 ■.-.<•
6.1 ;

3 73
74
AT *

17.7
17

. t : .

. . ■

4. .4

4,1 3,4.

7.0 39.

St

7.5

• — -

4

Ay.*

18
17.

3.1 17.
VfTb

5.0 1 ■•.
4.

1

7.v

6.'

. Ml,

6.

V?

5

7

Av.£

17.9
17

WIT

4,8 2a,

7.1 K.6

?#S 41.

6.5 38.
39.6

8 70

13
16

.4 19

3.0 19

9.1 38.

6,6 16.6

f • \ J »

1 33.4

7 71
64

17.4

13.1

4.

4. 1 ■ it.

. SO.

7,3 4 .

7.0 42.

T7.

7, v 43.

7.5 ja,

Hoa.l *nl 2 transferred to aerate l water at the oloj« of
the experiment mnlcrwcnt bo further re^enerati ,

71

TABL* 33 FXP^iyp^T 33
Regeneration in Lo* Oxygen

Jir

1

3

3

4

o

6

7

oo, 0C

par lit

er

■frXloUB

trace

1.0

1,9

.3

3.6

4.68

5.7

fcinimua

0

0*3

0.

1.0

1.8

.9

3.1

0*

0.

1,17

1.

.7

3.

4.1

Tilpolea

length

B3

7;;

82

85

80

83

8*

remor

i

.

22.

,

.

.7

33.0

Regenerate i

Date * be

•jun 3/^3/13

4/3

alight but not

measurable.

4/K)

4, ;

.

4,3

5.

.

3.8

.

$

17.

13,

MM

,7

6.

r.

.

4/15^

4.

4,8

5.5

.

.1

S.l

7.6

f

30,

34,

.

35.6

1.

39.

34.5

)

4,3

5.5

6.7

6.9

6.

7,6

8.5

*

*1.3

34,8

30.

1 .

i «

.7

• .5

3

5,3

5,5

6.9

7. 1

.3

7.6

3.4

*

33.

27.5

11.

31,

.

36.7

33.

72

$3 KXPSROTf si
tion In Low 0xy*en

Jar 13 3 4^6

oo .0<-
p*x litar

m»*l«iutt .33 8,76 4.6 8.3 1 .

a.lnlKum .13 0.5 1.9 .3 5.0 7*1

**«r. . 1.5 9.0 4, 43 7.1 8.38

TvinolM

l*n»-*h 71 73 78 73

1?.7 17. 13.3 18.. 19.

Re -jen« rated
Dato - bcT'iri 10/33/17

10/ - - 0,3

11/1

* 5.

11, - • 1.4

J 9.1

11/7

£ 1.7 14.

11/10 i5

£ .6 16.

11/13 r.s 1,

' .8 10,6

11/14 Boll«r toiled. ■»*

4.0
32. G

i

.0 8.8

• 16.

12/13 dS&J

0.8

0.
3.

b.O

m «•

1,1

b.o

0.
ft.

1,3
7.

4.6

dead

1.1

1.5

1.0

6.0

3.

19,

3.7
30.

3*0

4.

^3.9

33.

4.

34,

4.2

i •

1 . 1

re4

to boeofta

••Aerated.

5.

5.7
.8

5.0
1,7

8.

8.0
44.

43.

«•

54.

3.0
44.

iti

7^

TABIT 34

» FTPFPllfFKT 34

Effect i

of Temperature on Regeneration

t.C,

,Len

Length

Length Regenerated in wm.

in on.

remove i

BefTUn 3/6/18

In a»»

3/1C

B/1J

1 3/17

1 3/31

3/;6

3/30

4/6

II

n

X . ■•■

3.0

-.

8,0

8.0

8.0

to

63

16

1.1

3.8

5. .

7.0

7.3

7.

7.5

1

14

l.l

.

5.0

7.0

7.0

7.

7.4

10

tOtTil

7"7"

77

irKr

9 .. •

77"

5.6

.777

77

■777

t

7,3

19.

19$

•

45.9

43.

50.

14

61

15

i.6

1.1

3.4

3.

4.4

54

16

0*0

1.0

♦

3.7

4.

5.0

ea

14

0. 1

1.6

,

,fc>

1 *

M

totU

15
OCT

77

777

'ictrr

1TTT

177"

hi

■
• -

i

.

7*1

if

.9

.

.

0

16

(T)

to

55

16

Ho Re

-.it ion

4

6,
60

2

14.5
15

4/16

(T)

teti

4/30

4/

5/4

If

6.0

3.0

.0

8.0

8.

to

7.5

7.8

7.6

7.8

7.8

ai

7.6

7.6

7.7

7.7

7.7

•

-i*.

7~

6.0

4c..

49.

.

49.

.

14

6.4

6*5

C.5

6.5

S. 5

5.5

5,5

.5

5.7

5.8

6.0

6,0

6.

6.

6,0

8.7
• 77"

6.6

,77"

.777

~7"

41.

41.

41,

4J3.3

42.

0

3.

4.8

5.0

6.0

6.2

to

no regeneration

dead

4

4.1

5.3

.

6.3

6.5

no re-ener^t:

77*"

irrr*

iCTT

i~77

7T

34.6

,

34.4

40.

43.

(T) ■ Transferred to temperature 19 to 31 de^reee.

6
I)
to
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o
o

CO
C

o

•H

cd

■P

C
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c
o
o

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CO

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f-t

M

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03

M

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a

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P.

bD
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7S

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a
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a5

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c
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o

bO ©

C S

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>» 4*

M

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Graph showing $ regenerated, daily C$3,
time to regenerate 7.5^ and f> of normal
CO3 produced in increasing concentrations
of HC1.

Ordinate = cc.O.OlN HC1 in 300 cc,
Abscissa for $ regen. = f> removed,
for CO3 daily * cc.O.OlN H3CO3.
for 7,.5# regen. = time in days.
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of tables 31 and 23.

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Graph ahowin- the tirc© require! to regenerate

30> of the a&ount remoroi In 4«oreaelh& aonoen*

t rat tone of oxygen,

Orilnate • oo» oxygen x>«r liter*

Abeoieea ■ tine in lays.

From data of Table 33,

112

rtg« .

Graph •aftviag ra^anarvttlon in decrt&ainc o on cant rations

of fter the *at ar ha4 baen aerate .

Heavy perpendicular lino oarka time of entrance of

oxygen.

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1 liter of the water.

Frow lata of Tab la 33»

113

1 I

VITA

Klnna Xrrnetina Jewell •*?• born no-»r Inring, Kansas, February 9,
1892. She reoelved her secondary education In the public high so^ool of
Irving, JCanaas. In September 1910, aha enteral Colorado College, from which
institution aha received tna legree A.B. in 1914. Her major nae Biology.
In the fail of 1914, she enteral the Graduate Sohool of the Cnivereity of
Illinois as a Scholar in Zoology, and tho apring of 1915 aho received tha
degree M. A. froa tha sane institution. Tha ysar* 1915-1916 and 1917-1918 *ere
apant as n Gro^unta Fello* in tho Daparts»r.t of Zoology of the University of
IlllriOia, sni tha year 1916-1917 <*a a 3radu.-*ta Aosiat-uit. The samara r of
1916 woe apont at tha Pugent 8o«M Biological itfttltft, Sllea Jawall la a
•saber of Signa II an J Iota Sigma Pi.

Pu'ollcstione

1916. Cylindroteenia American? Hov. Spac. Froia

tha Crioket Frog. J Our.. Paraaltui..,

It 141-199.

1918. The Occurrence of Sichanaee in tho Digest lvs

Tract of In verteb rates. Jour .Biol.
Cheta., 33; 161-167. By Minn* K. Jawall
an J BafNUPd B. Lawia.

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