Radiation and gravitational effect on wasps and shrimp - animal stud

Grosch, D. S.

English

NASA Technical Reports Server

b 
STATUS REPORT on NsG-678 
UTILIZATION OF HABROBRACON AND ARTEMIA AS EXPERIMENTAL MATERIALS I N  
- 
BIOASTRONAUTIC STUDIES 
Prepared by D. S. Grosch 
Genetics Department, N. C.  S t a t e  Universi ty  a t  Raleigh 
Period Covered: January through June 1965 b -  
In t roduct ion  
During t h e  second s i x  months of this p r o j e c t ,  t he  combined e f f e c t s  of 
r a d i a t i o n  and fo rces  higher  than one g r a v i t y  have been inves t iga ted  on t h i s  
campus using t h e  wasp Habrobracon. 
and 19th  simulated space f l i g h t  tests a t  Moffet F i e l d ,  Cal i forn ia .  
Also w e  have pa r t i c ipa t ed  i n  the  Apr i l  13 th  
Our e f f o r t s  wi th  t h e  b r i n e  shrimp A r t e m i a  have been concentrated on 
s tandard iz ing  r ea r ing  i n  a chemically def ined environment. 
Habrobracon Experiments 
The Combined Ef fec t s  of Radiat ion and Cent r i fuga t ion  
I n  these  inves t iga t ions  w e  use a motor designed f o r  long continuous 
opera t ion  f i t t e d  with a head machined t o  hold capsules  of wasps. 
f o r  a f e w  minutes a t  a speed providing a fo rce  of 1,OOOg w a s  a treatment too 
Cent r i fuga t ion  
severe f o r  a l l  s t ages  o f - t h e  b racon id - l i f e  cycle .  
of a d u l t s  surv ive  500g f o r  as long as 24 hours. However, few eggs w e r e  l a i d  by 
females subjected t o  the  experience and h a t c h a b i l i t y  w a s  poor. Upon d i s sec t ion ,  
d i s turbances  of abdominal contents  were evident .  These included pos t e r io r  l o c a l i -  
za t ions  of u r a t e  cells ,  distended hindguts,  and enlarged egg sacs f i l l e d  with 
amorphous material with t h e  yellow color of degenerated eggs. 
O n  t h e  o ther  hand, t he  majori ty  
Af t e r  the  lower dose of 250g for 24 hours,  most of t h e  females l a i d  eggs 
and t h e  majori ty  of t he  eggs hatched. 
scale #mbination experiment a t  t h e  rate of revolu t ion  providing us with 250g. 
Our Co 
hour exposure. 
Therefore,  w e  decided t o  run the  f i r s t  f u l l  
gamma source de l ivered  30r p e r  hour t o  provide a t o t a l  of 720r f o r  a 24- 
The experiment w a s  repeated three times. During each r e p l i c a t i o n  samples of 
8 females w e r e  used f o r  each of t he  four ca tegor ies :  (1) cent r i fuged ,  (2) i r r a d i a t e d ,  
(3) cent r i fuged  and i r r a d i a t e d ,  (4) con t ro l .  S i g n i f i c a n t  evidence of po ten t ia ted  
damage from the  combination of treatments w a s  obtained i n  only one of t h e  th ree  sets 
of experiments. 
records f o r  e i t h e r  r a d i a t i o n  o r  cent r i fuga t ion .  
t he  lik of the  female is impressive. 
e f f e c t s  were most obvious only on t h e  days when r a d i a t i o n  induced low egg 
depos i t .  
a t i o n .  D i f f e ren t i a t ed  oocytes and undi f fe ren t ia ted  oogonia, r e spec t ive ly  a f t e r  and 
befpre  t h e  sensit ive period were l e s s  s e n s i t i v e .  
Both egg production and h a t c h a b i l i t y  w e r e  decreased below t h e  
The cumulative d a i l y  evidence fo r  
I n  the  o ther  two experiments t he  combined 
This  s e n s i t i v e  period corresponds t o  m i t o t i c a l l y  a c t i v e  c e l l  d i f f e r e n t i -  
Values fo r  egg depos i t  and 
https://ntrs.nasa.gov/search.jsp?R=19650018937 2020-03-17T01:29:33+00:00Z
2 
h a t c h a b i l i t y  a f t e r  a combination of treatments were genera l ly  intermediate  t o  
r e s u l t s  with e i t h e r  cen t r i fuga t ion  o r  i r r a d i a t i o n  alone. An apparent supe r io r i ty  
i n  performance during the  l as t  f i v e  days of l i f e  f o r  t he  females i n  one experi-  
ment can be viewed with skept ic ism i n  the  l i g h t  of t h e i r  being the  last  few 
females a l i v e  a f t e r  t he  majority of t h e i r  sisters making up the  t r ea t ed  sample 
had died.  I n  general  w e  f e e l  t h a t  death came r e l a t i v e l y  e a r l y  a f t e r  cen t r i -  
fuga t ion  and the  experiments need t o  be performed a t  s t i l l  lower rates of revolu- 
t i o n  and lower forces .  A tendency of a d u l t  females t o  o r i e n t  d i r e c t l y  opposi te  
t o  g rav i ty  (or  t o  appl ied force  i n  excess of grav i ty)  renders  t h e  abdomen and 
the  organism vulnerable .  ' I n  addi t ion t o  constant  speed of r o t a t i o n  these types 
of experiments include acce lera t ion  t o  and dece lera t ion  from t h a t  level. 
Simulated F l i g h t  P r o f i l e  and Radiation 
The Habrobracon experiments i n  simulated space- f l igh t  w e r e  designed t o  test 
t h e  r a d i a t i o n  subassembly and prototype packages under four separa te  condi t ions.  
These condi t ions were (1) the  simulated f l i g h t  p r o f i l e ,  (2) the  i r r a d i a t i o n ,  
(3) t h e  combined e f f e c t s  of t he  p r o f i l e  and i r r a d i a t i o n ,  and (4) the  non-profile- 
non i r r ad ia t ion  cont ro l .  
D r .  R. C .  von Bors te l  of the  Oak Ridge National Laboratory leads  a group 
pr imar i ly  concerned with packaging and assessment of gene t ic  damage t o  metaphase 
and prophase oocytes and spermatozoa f o r  t he  f i r s t  t h ree  days a f t e r  treatment 
(o r  recovery from a space satell i te) .  
f o r  t h e  remainder of t h e i r  l i v e s ,  about 20 days. 
fecundi ty  and f e r t i l i t y  and record l i f e  span. 
Subsequently,we follow the  same animals 
W e  study the  pa t t e rns  of 
Four pos i t i ons  w e r e  a v a i l a b l e  i n  each module. Females were placed i n  two 
pos i t i ons ,  males i n  t h e  t h i r d  and Toshiba g l a s s  rods i n  the  fourth.  
time f o r  t h e  Apr i l  13 test w a s  66 hrs.  20 min., and f o r  t he  Apr i l  19 test  66 hrs .  
40 min., with the  expectat ion of del iver ing doses i n  the neighborhood of 4000, 
2000, 1000 and 500r t o  t h e  modules i n  t h e i r  var ious  pos i t ions .  
measured dose f e l l  sho r t  of t he  nominal dose and the  r a d i a t i o n  i n t e n s i t y  f o r  the  
p r o f i l e  experiment proved t o  d i f f e r  from t h a t  of t he  non-profi le  experiment. A 
r e p o r t  dated 15 May 1965 from D r .  von Bors te l  t o  D r .  G. Dale Smith, Experiments 
Mgr ., B i o s a t e l l i t e  P ro jec t ,  NASA Ames Research Center, gives a d e t a i l e d  d iscuss ion  
of t he  dosimetric d a t a ,  along with conclusions and recommendations. 
I r r a d i a t i o n  
However, t h e  
Wasp su rv iva l  w a s  good except i n  the  nominal l O O O r  p o s i t i o n  where wasps 
w e r e  pulver ized during t h e  simulated re -en t ry  v i b r a t i o n  t e s t i n g .  
t h e  holding bracket  is  expected t o  minimize whipping of t he  package. 
Strengthening 
I n  both the  Apr i l  13 th  and 19th experiments a typ ica l  r a d i a t i o n  induced 
v a l l e y  w a s  obtained f o r  fecundi ty  a t  nominal doses above 500r. This v a l l e y  
occurr ing a t  t h e  end of t h e  f i r s t  week r e f l e c t s  t h e  r a d i o s e n s i t i v i t y  of d i f -  
f e r e n t i a t i n g  oocyte-nurse-cell  nes t s  and ind ica t e s  t h a t  t he  doses de l ivered  were 
adequate f o r  studying modifications i n  the  p a t t e r n  of egg production. 
obtained by p l o t t i n g  egg depos i t  aga ins t  days w i l l  no t  be presented here.  
show no unusual pa t t e rns .  
The curves 
They 
3 
The cumulative e f f e c t s  on the s e n s i t i v i t y  of developing eggs can be 
summarized as the  average t o t a l  eggs produced per female. These sums are shown 
i n  Table 1 f o r  t he  most s i g n i f i c a n t  period following the  Apr i l  13th experiment. 
The per iod chosen avoids (a) eggs deposited t h e  f i r s t  t h ree  days which w e r e  
d i f f e r e n t i a t e d  oocytes a t  t he  t i m e  of treatment and (b) eggs deposi ted la te  
enough t o  be derived from und i f f e ren t i a t ed  oogonia. 
The t o t a l s  r e in fo rce  t h e  impression that v i b r a t i o n  alone has l i t t l e  e f f e c t  
upon ov ipos i t i on  records.  
w a s  produced i n  combination with nominal r a d i a t i o n  dose 4000r. 
dosimeter 2.30 K r  measured f o r  the combination treatment i s  less than t h e  2.59 K r  
found f o r  t h e  r a d i a t i o n  alone. However, t hese  d a t a  are the  only ind ica t ions  of 
synergism obtained. A l l  o the r  egg production summations appear t o  r e f l e c t  the 
6% average lower r a d i a t i o n  dose i n  the combined r a d i a t i o n  and v i b r a t i o n  p a r t  of 
t h e  experiment. Table 2 gives  egg production summaries f o r  the  r a d i o s e n s i t i v e  
per iod and the  subsequent period ( u n t i l  s e n i l i t y )  f o r  t he  A p r i l  19th experiment. 
On the  other hand, a s t r i k i n g l y  low t o t a l  of eggs 
Actual ly  by 
Hatchabi l i ty  records appear t o  r e f l e c t  merely t h e  r a d i a t i o n  dose. The 
V a r i a b i l i t y  between 
combined treatment i n  which 6% less r a d i a t i o n  w a s  de l ive red  c o n s i s t e n t l y  shows 
higher h a t c h a b i l i t y  than the  r a d i a t i o n  exposure alone. 
p o s i t i o n s  is small and f o r  s impl i c i ty  of summarization r e s u l t s  f o r  t he  two female 
p o s i t i o n s  are pooled. On t h i s  bas i s  the  nominal 4000r dose f o r  t he  f i rs t  t h r e e  
days o f  each experiment shows the  following h a t c h a b i l i t y  r e s u l t s  i n  per  cent :  
Radiation Radiation & Vibra t ion  Vibra t ion  
Expt. 1, Apr i l  13-16 64.50 72.26 98.30 
Expt. 2 ,  Apr i l  19-22 65.39 69.14 97.30 
Control  h a t c h a b i l i t y  w a s  94.28 
For t h e  eggs of Table 1, the  highest  nominal dose 4000r c h a r a c t e r i z e s  t h e  
p a t  tern: 
Radiation Radiat ion & Vibra t ion  Vibra t ion  
52.27 55 .OO 42.72 
These r e l a t i v e l y  low va lues  m u s t  be viewed i n  t h e  l i g h t  of c o n t r o l s  which had 
decreased t o  57.97% hatched during t h i s  period. 
treatment r e s u l t e d  i n  a higher value than r a d i a t i o n  alone. 
Nevertheless,again t h e  combined 
Hatchabi l i ty  f o r  t h e  eggs o f  Table 2 are shown i n  Table 3 f o r  t h e  h ighes t  
nominal dose (4000r). With standard e r r o r s  of t he  order  of 9% d i f f e r e n c e s  between 
t reatments  are n o t  s i g n i f i c a n t .  Although a v a i l a b l e ,  h a t c h a b i l i t y  d a t a  f o r  the 
lower r a d i a t i o n  doses w i l l  not be presented here.  Differences from c o n t r o l  va lues  
are even smaller f o r  t h e  lower doses. 
4 
Discussion of the Current Emphas is  in Habrobracon Experiments 
I n  space- f l igh t  biology problems of i n t e r p r e t a t i o n  arise because l i v i n g  
material is  subjected t o  a number of f ac to r s  whose a c t i o n  on he red i t a ry  
s t r u c t u r e s  is  not y e t  adequately understood i n  i s o l a t i o n ,  l e t  alone i n  combination 
with chronic  r a d i a t i o n  exposure. 
accomplished i n  the  laboratory but the f a c t o r s  of v i b r a t i o n  and of acce le ra t ion  
t o  fo rces  i n  excess of g rav i ty  can be inves t iga ted  i n  cont ro l led  experimental 
fashion. Cytological  inves t iga t ions  of t he  e f f e c t s  of cen t r i fuga t ion  have 
t r a d i t i o n a l l y  employed germinating seeds (Kostoff,  1935; Cytologia 8:420; Sax, 
1943; P.N.A.S. 29:18; Wolff and v .  Borstel ,  1954. P.N.A.S. 40:1138.) Attempts 
t o  produce gene t i ca l  changes i n  insec ts  have not  been reported u n t i l  r ecen t ly  
(Reddi, 1963. Nature 198:316), possibly because r e s u l t s  have e i t h e r  been in- 
cons i s t en t  or  negative.  
Extended per iods of weightlessness cannot be 
Reports from the  U . S . S . R .  have been incons is ten t .  I n  crossing-over inves t i -  
ga t ions  with Drosophila an e f f e c t  was observed i n  one f l i g h t  bu t  not i n  two o thers .  
By turn ing  t o  simulated f l i g h t  experiments it w a s  poss ib le  t o  show t h a t  a c c e l e r a t i o n  
t o  4000g had l i t t l e  e f f e c t ,  whereas v i b r a t i o n  had d e f i n i t e  consequences. 
c a t i o n s  a rose  when combination experiments were set up using gamma rays.  
doses of l O O O r  and 2000r v i b r a t i o n  apparently had opposi te  e f f e c t s  from c e n t r i -  
fugat ion.  (Parfenov, 1964. Moscow Akademiya Nauk, v o l .  2).  
Compli- 
With 
I n  t h e  Vostok satell i tes '  Tradescantia experiments, chromosomal abe r ra t ions  
were t raced  t o  dynamic f a c t o r s  accompanying launch and recovery of t he  space 
c r a f t  but  not  t o  the  state of weightlessness.  The smallest y i e ld  w a s  obtained 
i n  material wi th  the  longest  experience of weightlessness;  t h a t  i s  i n  material 
f ixed  p r i o r  t o  landing. (Delone e t  a l .  1964, NASA TT F-8896.) 
S imi l a r ly ,  the  i n v i a b i l i t y  of Drosophila eggs from a d u l t s  c a r r i e d  by the  
Vostok 3 and 4 spacecraf t  is not  considered t o  be due t o  weight lessness .  Nor w a s  
ion iz ing  r a d i a t i o n  alone a s ign i f i can t  f ac to r .  
seems t o  support  t he  idea of e f f e c t s  due t o  v i b r a t i o n  but  a l s o  concern is  expressed 
about l a x  temperature con t ro l  during the  recovery period. (Parfenov, 1964. NASA 
TT F-8898.) 
An increase  i n  dominant l e t h a l s  
Fortunately Habrobracon a r e  not as temperature s e n s i t i v e  as Drosophila. 
Nevertheless, t h e  absentmindedness of an  Oak Ridge ind iv idua l ,  not of D r .  von 
B o r s t e l ' s  g roupdelayed  our procurement of t he  wasps from the  second experiment 
and could have contr ibuted t o  the  r e l a t i v e l y  low con t ro l  va lues  of Table 3 .  
W e  f e e l  it des i r ab le  t o  g ive  some a t t e n t i o n  t o  v i b r a t i o n  experiments i n  
add i t ion  t o  cen t r i fuga t ion  experiments. 
A s  a l ready  s t a t e d  changes a r e  planned i n  the  simulated space f l i g h t  f i t t i n g s .  
add i t ion  the  modules i n  each package can be a l igned  i n  a row p a r a l l e l  t o  t he  
backsca t te r  sh i e ld  r a t h e r  than i n  a rectangular  pa t t e rn .  
equa l i ty  of r a d i a t i o n  d i s t r i b u t i o n  but a l s o  d i s t r i b u t i o n  of v i b r a t i o n a l  e f f e c t s .  
D r .  von Bors t e l ' s  r e p o r t  goes i n t o  more d e t a i l  on package modif icat ions.  
Much of t h i s  can be done on our campus. 
I n  
This may not  only improve  
5 
A r t e m i a  Experiments 
The formulation of a superior  synthe t ic  sea water has made poss ib le  the  
r e a r i n g  of A r t e m i a  i n  a chemically defined environment anywhere i n  t h e  world. 
W e  have demonstrated t h i s  using the  commercial product, " Ins tan t  Ocean", 
successfu l ly  employed by inland aquaria f o r  the  maintenance of l a rge r  marine 
organisms. However, we f ind  t h a t  increasing the  s a l i n i t y  by adding 50g of N a C l  
per  liter is important here  as w e l l  as when using n a t u r a l  sea w a t e r .  During 
t h e  Spring Semester two sets of 15 p a i r  matings were followed i n  I n s t a n t  Ocean 
so lu t ions .  Measured i n  days the average l i f e  span without s a l t  w a s  20.47 f o r  
males and 29.86 f o r  females; with s a l t  73.67 f o r  males and 62.13 fo r  females. 
Furthermore the  addi t ion  of N a C l  increased the  average number of broods p e r  
female from 3.73 t o  10.13 and t h e  average number of of fspr ing  p e r  p a i r  from 
169.6 t o  1423 .O. 
W e  look forward t o  using A r t e m i a  as w e l l  as Habrobracon fo r  studying the  
I n  a l l  p robab i l i t y  A r t e m i a  holds promise b io log ica l  e f f e c t s  of space f l i g h t .  
as t h e  most v e r s a t i l e  and usefu l  b io logica l  dosimeters.  
Personnel of P r o j e c t  
Leader: D. S. Grosch, Professor  of Genetics 
Research Ass i s t an t s  (Half time during academic year) :  
James C-H Lin,  Graduate Student 
Roger H. Smith, Graduate Student 
‘II . .- 6 
Table 1 
The per female average braconid egg production from the  t h i r d  t o  the  t e n t h  day 
following simulated space f l i g h t  (April 1 3 ,  1965) .  
Nominal Pos i t i on  Radiation Radiation & Vibra t ion  Controls  
i n  r 
Dose Vibrat ion 
4000 B 45.6 29.0 88 .O 67.6 
C 50.9 31.2 90 .o 107.5 
2000 B 43.6 64.7 74.3 105.4 
C 51.6 66.8 85.2 89 .O 
1000 B 71.4 73.2 
C 46.5* 53.1 
83.3 
93.0 
500 B 7 1  .O 64.6 82.6 71 .O* 
C 77.3 75.3 92 -4 97.2* 
* 
Five days only. 
Table 2 
The per  female average braconid egg production f o r  the  per iods shown following 
simulated space f l i g h t  (Apri l  1 9 ,  1965).  
Days 4 through 9 
Nominal Pos i t i on  Radiation Radiat ion & Vibrat ion Controls  
i n  r 
Dose Vibrat ion 
4000 
2000 
1000 
5 00 
Days 10 through 17 
4000 
2000 
500 
B 34.30 61.97 94.22 134.60 
c 35.80 59.97 97.86 106.90 
B 53.41 79.25 99 .oo 93.67 
C 68.20 71.87 82.80 98.45 
B 69.20 
c 97 .oo 
B 78 .OO 89.45 88 27 104.33 
c 77.14 98.50 98.67 80.78 
B 42.75 108.83 97 -83 147.42 
C 104.75 119.67 86.62 65.83 
B 83.25 93 -50 147.42 
C 104 55 93.17 103.09 139.09 
B 81.50 127.03 141.04 
C 108.04 117.33 109.80 91.93 
. t  
“r .*L.. 
c 
7 
Table 3 
Hatchabi l i ty  of the  eggs of Table 2 f o r  t he  nominal dose of 4000r. 
pos i t i on  d a t a  w e r e  pooled f o r  summary. A l s o  th ree  r a t h e r  than two per iods 
a r e  shown. 
B and C 
Days Radiation Radiation & Vibra t ion  Control 
Vibration 
4- 7 36.91 42.39 35.69 52.37 
8-11 29.92 27.39 37.65 47.58 
12-18 26.15 29.25 38.65 37.33 


Utilization of habrobracon and artemia as experimental materials in bioastronautic studies status report, jan. - jun. 1965

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