The insecticidal action of sulphur
by G D Green
A THESIS Submitted to the Graduate Committee in partial fulfillment of the requirements for the
Degree of Master of Science in Entomology
Montana State University
© Copyright by G D Green (1932)
Abstract:
no abstract found in this volume TK2 IlISSC TI Cl DAL ACTIOH OF SULPHUS
by
0 . D. GEBasr
A THSSIS
S u b m itted to th e G rad u ate Committee i n p a r t i a l f u l f i l l m e n t
o f th e re q u ire m e n ts f o r th e D egree o f
M a s te r o f S c ie n ce i n Entomology
a t M ontana S t a t e C o lle g e .
Approved:
I n C harge o f M ajor Work
c,
S f c + +++-L.
Chairm an Exam ining Committee
Chairman G raduate Committee
Bozeman, M ontana
J u n e , 1932.
SWTf
1,
M37f
TABLE OF COHEETS
Page
INTBOIXJCTI ON..........................................................................................
I
ACKNOWLEDtMJOT.......................................................................................
I
EEVIEW OF LITEEATUEE............................................................................
I
EXPBBIMEOTAL METHODS AND EESULTS ......................
15
The Production o f Hydrogen Sulphide by
Sulphur Treated I n s e c ts ......................................
15
The E ffect of Tenperature on the B eactlo n ...............
19
The E ffect of Hydrogen ion Concentration
on the B e ac tlo n .....................................................
Hydrogen Sulphide Production by Lime-Sulphur . . . .
20
The Toxicity of Sulphur to some S p e c ie s...................
20
The Belationship of the Glutathione Content
of a Species to the A bility o f that
Species to Produce Hydrogen Sulphide
from S u lp h u r.........................................................
21
The Effect of Sulphur on the Catalase A ctivity
of Melanoulus b lv itta tu s . ...............................
'23
The Toxicity of Hydrogen Sulphide to Insects . . . . .
. 24
DISCUSSION AND CONCLUSIONS................................................................
2S
SUMMAEI .................................
30
LITERATURE CITED ............................................................ . . . . . .
31
43085
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THE IHSEOTICinAL ACTION OE SULPHUR
INTBODUGTIOH
Sulphur alone or In combination with other substances was
among the f i r s t m aterials recommended fo r use in the control of certain
types o f Insects and p lan t diseases.
These m aterials are today among
the most important insecticides and fungicides.
In sp ite of th is wide
and long continued use, and the great amount of woik which has been
directed along th is lin e , no d efin ite conclusion can be drawn as to the
toxic property o r how the toxic effect i s produced.
r'
The w riter has made an e ffo rt to determine the so le of hydrogen
sulphide in the action o f sulphur when applied to In sects.
Besults of
th is work are presented in th is paper, together with an attempt to
review some of the view points expressed by other workers as to the action
of sulphur and lime-sulphur solution as in secticid es or fungicides.
ACKNOWLEDGMENT
The w riter g rate fu lly acknowledges h is indebtedness to Doctor
JL L. Strand fo r proposing th is investigation and fo r h is in te re s t,
encouragement and many helpful suggestions during i t s progress.
BBVIEW 0? LITEBATUEE
The use of sulphur in cleansing wool and clothes of moths was
practiced by the ancients, being mentioned in the Old Testament. Erasmus
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3-
Darwin (1800) re la te s tba. discussion on the use o f a mixture of lime
and sulphur in the control of aphids, m ites, and scales.
Ilftnental
sulphur, usually in mixtures with in e rt substances as d ilu en ts, has been
in continued use since the early times fo r the control of various insects
and fungi.
At the present time i t i s used as a specific remedy fo r
Infestations of cotton flea-hoppers, many species o f m ites, etc.
The use
of lime-sulphur and compounds of other substances and sulphur does not
seem to have been so consistent.
Barium-sulphur is used to some extent,
but i t s use is not common. A combination of potassium and sulphur, known
as liv e r of sulphur, has been used in England quite extensively against
certain fungus diseases.
By fa r the most extensively used sulphur-containing m aterial is
lime-sulphur.
I ts p rinciple use before 1889 was in stock dips, but
beginning with th at year i t has been used as a spray fo r the control o f
scale in se c ts.
As the best known remedy fo r San Jose scale, i t s use
accompanied the rapid spread of th is insect over the United S tates.
Tor
scale in se c ts, alone, i t has been larg ely superseded, during the la s t
few years, by o il sprays of various kinds.
However, where there are also
fungus diseases to be combatted, lime-sulphur i s generally used.
Hundreds of experiments have been performed in the e ffo rt to
determine the value of th is m aterial in controlling in se cts.
Most of them
have been directed toward finding the most effectiv e proportion of lime
and sulphur to use, the proper length of time to b o ll, the best time to
apply, and the correct d ilu tio n to use.
The conclusions were drawn from
re su lts of p ra c tic a l fie ld experiments.
Comparatively l i t t l e study had
been made from a chemical o r physiological standpoint u n til about 1905.
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InveetlgationB on the chemical composition, together with
te s te as to the effective component have v e rifie d the methods and
foxmulae recommended fo r the preparation o f lim e-sulphur.
Standard
directions, fo r the home preparation of the concentrate, advise to ilin g
one hour and use of the Ingredients in atout the ra tio o f one pound stone
lime, two pounds flowers o f sulphur and enough water to make the finished
product one gallon.
Trom a chemical standpoint, th is gives the g reatest
content of the components which have proven most valuable.
A considerable amount of woik has been done in try in g to dis­
cover the toxic property o f sulphur and o f lime-sulphur sprays. Many
confusing conclusions have been drawn.
The cause of the g reater p art of
th is confusion i s lik e ly experimenting under d ifferen t conditions, mis­
in te rp retatio n of re su lts, and basing conclusions on In su fficien t data.
Most to x ic ity te s ts of the components shown to be associated with sulphur
o r lime-sulphur have been applied to fungi.
As there is probably no
essential difference in the toxic action o f these m aterials on fungi and
Insects, the lite r a tu r e on both are included.
The f i r s t comprehensive study o f the chemical composition of
lime-sulphur solution seems to have been made by Hayrood (1905).
He
+.Virmght tfgBk the primary reaction between lime and sulphur in aqueous
solution to be as follows:
3 Ca (OH)2 *-12 S-= Ca S2O3 + 2 Ca S5 -f 3 H^>.
As th is leads to lees thiosulphate and more pentasulphlde than analyses
show to be present, he said th a t probably one o r both of the following
secondary reactions occur:
3 Ca (OH)2^ g S = Ca S2O3 + 2 Ca S3 + 3 H2O
Ca S5 + 3 0 = Ca S2O3 + 3 S.
and
I f to llin g continues, more thiosulphate I s formed a t the expense of
pentasulphide.
In solution, thiosulphate changes to sulphites and these
in turn to sulphates as follows:
Ca S2P3 * Ca S 03't'S
Ca S O3-K)= Ca S O4 •
These facts led Haywood to the conclusion that probably the lime-sulphur
sa lt wash i s formed as follows:
3 Ca (OE)^lZS = Ca S2O5 V-S Ca S5 4-3 H2O,
Ca 83+3 0 m Ca S2O5^ 3 8,
and th is sulphur V- Ca (OH)2 = Ca S2O5^ 2 Ca S3 4-3 HgO,
to a slig h t extent Ca S5O3
= Ca S O34-S
Ca S O34O z Ca S O4.
The presence of s a lt does not affect these reactions.
This author thinks th a t, in dry clim ates, when scales are
treated with th is wash, the excess lime loosens the scales and exposes
the insect beneath and th a t the calcium pentasulphide is decomposed thrombi
the various intermediate steps into free sulphur and calcium sulphite,
which are the effective agents.
In wet clim ates, rains would wash o ff
most of the thiosulphate and the in se cticid a l value would be diminished
in proportion to the amount washed o ff.
Haywood (1905), incidentally, re la te s a frien d , Mr. F. H. Pough,
as expressing the opinion th at the action of lime-sulphur was due to the
gradual oxidation of sulphur into sulphur dioxide and sulphurous acid,
which are the valuable constituents.
Haywood doubts the oxidation being
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rapld enough to be of value, and c re d its sublimed sulphur fo r the supposed
odor of sulphur dioxide on hot days.
T arter and Bradley (1910) believed that the polysulphides in the
solution were probably a mixture of the te tr a and pentasulphlde of calcium,
and th e ir analyses showed evidence o f a more stable poly sulphide, which
was thought to be, probably, the disulphide of calcium.
Biey decided th at
a considerable amount of the sulphur in the solution Ie very feebly
combined and fo r p ra c tic a l purposes might be considered as sulphur In
physical solution, and th at i t Is not necessary fo r the spray-to oxidize
in order fo r free sulphur to be deposited from I t .
They found carbon
dioxide to have a small but evident influence in the decomposition of
the polysulphides, probably as follows:
Ca S^+HgC O3 = Eg. 8 f C a C
.
These authors said th at the presence of hydrogen sulphide should
not be overlooked shen considering the immediate action of lime-sulphur.
Their conclusion is th a t lime-sulphur solution i s a complex system
subject to the influence o f many varying conditions.
In 1910, Ibrenan published an account o f work with liv e r of
sulphur.
As used a t th at time, liv e r of sulphur was not a standard
product, but consisted o f compounds of sodium as well as potassium.
He
found, in addition to sulph-hydrate, sulphides and polysulphides o f e ith e r
potassium o r sodium, in most cases free sulphur.
The oxidation products
showed l i t t l e o r no fungicidal properties, when trie d separately, in weak
0
solution, upon the spores of B atrv tis clnerea. Saturated solutions of
hydrogen sulphide o r free sulphur had absolutely no adverse effect upon
the germination of spores of th is fungus. He concluded from the re su lts
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of M b investigations that the most potent fungicidal agent in liv e r
of sulphur i s free sodium hydroxide and th a t potassium hydroxide was some­
what le s s valuable. He a ttrib u te s no more effect to sulphur as a dust
than to ordinary dust.
Results of a series of carefu lly executed experiments by Shafer
(1911) indicated that the -value of lime-sulphur as a ecalecide lie s in
i t s strong, p ersisten t reducing power, and i t s a b ility to soften the wax
about the margin of scale in sects.
Anld (1912) removed considerable hydrogen sulphide by passing
p u rified nitrogen o r oxygen throu^x lime-sulphur solution, the evaluation
increasing with rise in temperature. He thought the concentrated,
commercial product to contain, generally, the polysulphides, thiosulphate,
sulphite and sulphate of calcium, the poly sulphides and thiosulphate in
preponderance. He had no doubt th at most of the sulphur in the solution
i s in a very loosely combined condition.
T arter (191%), a f te r working on th is question fo r several years
and reviewing the conclusions of Shafer (1911) end others, enumerated
the following properties as the ones to which, in general, the
in se cticid a l action of lim e-sulphur seemed to be due.
( l) I t s power to
take up large amounts of oxygen; ( 2) i t s a b ility to soften the newly
secreted wax a t the margin of scale in sects, and (3) the amount of free
sulphur deposited in i t s decomposition.
According to Chapin (1916), ignoring unessential and
hypothetical intermediate compounds, the following reactions apparently
occur when lime and sulphur are bailed with water;
(1) 3 Ca (OH)2 -HES * 2 Ca S5+-Ca 8 ^ + 3 HgO,
(2) I 0 Ca S5+-3 Ca (OH)2 * 12 Ca S^-4-Ca
and (3)
Ca S4+S
«
Ca
3 H2Of
,
Beactlon 3 holds with excess sulphur and only in absence o f such excess
does equation 2 hold.
He thought a l l I Ime-sulphur solutions subject to
hydrolytic decomposition according to the equation:
Ca Sx-J-E H2O - Ca (0H)2+H2S+(X -l) Sf
the sh iftin g of equilibrium from l e f t to r l ^ i t increasing with ris e of
temperature.
In case of Ca Sg in the open a ir , where hydrogen sulphide
can escape the fin a l product i s as follows:
Ca Sg+-3 H2O ■ Ca S2Og+-3 H2 S.
Doran (1922) found th a t commercial lime-sulphur in dilutions
of one to ten did not prevent the germination of the conidia o f Tenturla
3
Inequalla in the absence of sulphides.
In a single experiment he obtained
absolute control of th is fungus when dusted with sulphur and exposed to
a ir , and none in the absence of oxygen.
A ris e in temperature or a longer
time of exposure Increased the to x icity o f sulphur in h is experiments.
Thatcher and S tre ete r (1925) suggest th a t elemental sulphur,
as deposited in the decomposition o f lime-sulphur o r applied in duets, is
a solid, no% toxic m aterial, i t s toxic effect being produced only a f te r
i t has been changed into some gaseous fo ra.
Conclusions of other workers,
mentioned, a ttrib u te d the fungicidal effect to sulphur dioxide produced
by oxidation of sulphur in moist a ir .
I t is reported th at other workers,
by use of pure sulphur dioxide, have fa ile d to verify these conclusions.
Two possible and differen t fungicidal e ffe c ts of sulphur c o n ta i n i n g
m aterials seemed obvious to these authors.
One i s the specific toxic
effect of calcium poly sulphide or sim ilar compounds which can exert th e ir
effect only so long as these m aterials remain as such on the tree and
against only the disease with which they come In contact.
The second
effect is th at due to the gaseous substances produced by the oxidation of
free sulphur which continues as long as there i s enough sulphur remaining
on the foliage to produce & su fficien t amount of the toxic gas to k i l l She
p a rtic u la r pest in question.
In an ordinary meeting of the Association of Economic B iologists,
October 20, 1925, a long discussion on the fungicidal action of sulphur was
carried on.
The following points are recorded from th a t discussion,
Anon (1926);
Professor B.T.P.Barker, in discussing the work a t Long Ashton,
said: a t the outset i t was recognized th a t atten tio n could not be confined
to the action of uncomblned sulphur.
Hence, the action of spray flu id s
of the poly sulphide type were examined and the conclusion arrived at th at
th e ir protective fungicidal action must be a ttrib u te d to the sulphur
which i s rapidly produced by the decomposition of the polysulphides a f te r
the application of those sprays to the p la n t.
Varied experiments
demonstrated th a t the toxic effect could be exerted across a space.
When a ir , in a greenhouse treated with a coating of sulphur on the hot
water pipes, was f ilte re d through cellulose pads o r passed through bent
glass tubes, i t was found to be non-toxic and p a rtic u la te sulphur was
deposited on the pads and in the tubes.
so trea ted was toxic.
Air, from the same source, not
The resu lts were Interpreted to mean th at sulphur
trav els through the a i r In p artic u late and not in gaseous form, and th at
i t Is In the form of elemental sulphur and not a compound.
Three altern ativ e
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views were deduced ae to the action of th is p artic u late fora.
(I) The
p o ssib ility that p artic u late sulphur is dissolved in the film of moisture
surrounding the fungus.
p artic u late sulphur.
This view was dispersed hy the in so lu b ility of
(2) Sulphur d ire c tly toxic in p a rtic u la te sta te , a
view unsupported by p o sitiv e evidence.
(3) The th ird a ltern ativ e action
presented was th at of sulphur compounds formed from p a rtic u la te m aterial.
The substances given consideration under the th ird viewpoint
were sulphur dioxide, sulphur tr io rid e, sulphuric acid, pentathlonlc acid,
hydrogen sulphide, and other sulphides.
No evidence of the formation,
in adequate strength, of sulphur dioxide, sulphur trio rid e , or sulphuric
E xperim ent S ta tio n
acid eradicated these substances.
Long Ashton^work did not confirm the
pentathlonlc acid theory of Young, and the fact th a t Young found sulphur
toxic outside the lim its of hydrogen ion concentration stated as necessary
S. a I J
fo r the s ta b ility of pentathlonic^seemed enou^i to put i t out of account.
The production of hydrogen sulphide when green leaves are treated with
sulphur was mentioned.
Mr. T attersfield of Bothamsted considered th a t with respect to
the fungicidal action of the polysulphides, i t would d e fin ite ly appear as
i f the d irect action were due to a lk a lin ity and the action of the
polusulphides as such, but th at the prolonged effect and the action a t
a distance were in some way bound up with the sulphur p a rtic le s deposited
from the solution.
Goldsworthy (1928) believed the fungicidal properties of
polysulphide solutions dependent, a t le a s t in p a rt, upon the soluble
sulphide content.
He found sulphur to be deposited in germ tubes, and
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Bpores to be arrested In th e ir development, and concluded the le th a l
factor to he the oxidation of sulphide to sulphur.
This was thought to
destroy the poising m aterial of the oxidation-reduction system to a point
beyond which recovery i s impossible.
Goodwin and Martin (1928) found that by heating glass wool
f i l t e r s o r glass tubes and drawing a ir , from sulphur trea ted greenhouses,
through them that no p a rtic u la te sulphur was deposited and th at the a i r
did not lose i t s toxic property.
This proved th at the sulphur was gaseous.
The removal of the v o la tile agent by passage throu^i a cooled f i l t e r was
taken as proof that i t was n eith er sulphur dioxide o r hydrogen sulphide,
but was elem ental' sulphur.
Condensation of v o la tiliz e d sulphur on cooled
surfaces was given as the reason fo r the deposition of p a rtic u la te sulphur
mentioned by B aiter.
Boach and Glynne (1923) concluded, te n tativ ely , th at th io sulphuric
acid was the principal toxic substance associated with sulphur.
Other
acids normally associated with sulphur were found to have about equal
to x lc ltle s and th is was a ttrib u te d to the hydrogen ion concentration.
Williams and Young (1929) found ordinary ground r o ll and flowers
of sulphur to yield varying amounts of the toxic facto r, depending upon
the reaction, oxidation, temperature, and moisture.
oxidizing agents enhanced the to x ic ity g reatly .
The addition of
From te s ts with a l l the
acids known to be associated with sulphur, they report only the polythionic,
( te tr a and penta), acids to be toxic.
acids were non-toxic.
Samples of sulphur freed from these
The toxic facto r was said to be destroyed by strong
acids and a lk a lie s.
The main criticism s directed against the conclusions of
Williams and Young seem to be the following: ( l) Their fa ilu re to explain
-li­
the I r method of Iso latio n or preparation o f the pentathlonic acid used.
This, with the known d iffic u lty of preparing th is acid free from
im parities, and i t s variable s ta b ility under d ifferen t conditions, seem
to be taken as evidence th a t they could not be d e fin ite ly sure of the
compound with which they were working.
(2) Their statement that
pentathlonic acid was stable only within the pH lim its of U.2 - 5.4 and
then giving resu lts of fungicidal a c tiv ity of sulphur below a pH of 3.2
and 2.4.
According to DeOng and Huntoon (1929), the insecticidal action
of lime-sulphur solution i s due, f ir s t , to the reducing or oxygen absorbing
power of the sulphide, and secondly, to the action o f free sulphur.
Tucker (1929), by a series of experiments, found sulphur to be
deposited on the sides of containing vessels a t ordinary temperatures.
At. the temperature of b o ilin g water, only traces of hydrogen sulphide o r
the products of oxidation o f sulphur could be detected, leading to the
conclusion th at th is deposited sulphur was the re su lt o f condensation o f
sublimed sulphur.
The fa c t th at the increased ra te o f action with succeed­
ing rise s in temperature of 10%. corresponded with the ra te of physical
rather than chemical action strengthened th is conclusion*
In the control
of ce rtain plant diseases, the effectiveness of sulphur increases a t the
same ra te , with rise s of temperature, as the ra te of sublimation.
The
value of sulphur as a fungicide. Tucker thought, depended prim arily upon
the sublimation of sulphur as such.
The resu lts of various workers who have found sulphur to act
across a space confixm th is conclusion.
Further proof of the action of sulphur through a distance is
-1 2 -
d f
! rqt. H I
tbey\re s-olte o f McOregor (1927) with the tarnished bog, Lygos e ll sub.
These Insects, confined In screen cages 12 to 24 Inches above the gro-und,
over sulphur dost lib e ra lly applied to the surface of the s o il, at
temperatures of from 102??. to 116*?., died within 30 minutes.
The controls
sim ilarly treated , except the sulphur, suffered no m ortality.
O’Kane and Conklin (1930), from re su lts of laboratory te s ts ,
concluded that hydrogen sulphide Ie not given o ff from lime-sulphur solution
In su fficie n t quantities to account fo r the to x ic ity of th at m aterial.
They found no trace of sulphur dioxide being evolved.
Ho method of analysis, yet suggested, w ill show the concentration
of hydrogen sulphide In the Immediate v ic in ity of an Insect In contact
with sulphur or I Ime-sulphur solution.
The chances are th a t I t Is many
times g reater than analyses show, and I t cannot be lower.
Wllcoxon and McCallan (193°) found the to x ic itie s of sulphuric
and pentathlonlc acids to be equal, within experimental erro r, and hydrogen
sulphide to be 6 to 200 times as toxic as these acids under varying
conditions.
Water ex tracts o f sulphur gave no control.
The to x ic ity of
Consnerclal dusting sulphur, a f te r the removal of acids associated with i t ,
was fu lly as, high as the same m aterial without such treatm ent.
I t was
concluded th at pentathlonlc acid is not a facto r of Importance In the
fungicidal action of sulphur.
McCallan and Wllcoxon (1931) present a large amount of evidence
that liv in g tissu es in contact with sulphur are able to bring about i t s
reduction to hydrogen sulphide.
In th e ir te ste on 26 species of plants
of 16 fam ilies, hydrogen sulphide was produced In every case, there being,
however, marked v ariatio n in the ra te of production by d ifferen t species.
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Spore a from some 27 species of fungi produced hydrogen sulphide when
trea ted with sulphur, the r&te In th is case, also, varying with d ifferen t
species.
nature.
The effect o f_temperature indicates a reaction enzymatic in
They found th at, within the pH range of U.O to 8.0, the rate of
evolution-to he Independent o f hydrogen ion concentration.
Jl
concentration
of 0.2 mg. p er l i t e r of hydrogen sulphide was completely toxic to Venturia
Inaecroalis nwhere with B otrytls spp. of the clnera type, there was 3.8 percent
germination a t a concentration of 20 mg. per l i t e r .
The order of
se n sitiv ity of 8 species of fungi to hydrogen sulphide and to sulphur
was exactly the same.
There was a p o sitive co rrelatio n between the
sensitiveness of a species to hydrogen sulphide and i t s a b ility to produce
that gas when treated with sulphur.
To give a c le a re r presentation of the point of view o f McOallan
and Wllcoxon (1931) their"Discussion" sh a ll be quoted.
"In presenting
any theory to explain the fungicidal action of sulphur cognizance must
be taken of the following experimentally determined fa c ts.
Leaves and
spores of a l l species of p lan ts tested produce hydrogen sulphide when in
contact with sulphur.
The production of hydrogen sulphide by spores varies
according to the species and i s d irec tly proportional to the number of
spores.
The optimum temperature i s 359C* and in h ib itio n takes place a t
55eU. but the production occurs over a re la tiv e ly wide pH range.
The
tno-Hnmm ra te of production a t 30%. ie reached in about 3 hours.
Actual
contact between spores and sulphur i s not necessary as action w ill take
place through a collodion membrane, the hydrogen sulphide being produced
on the spore side of the membrane and not on the sulphur side.
also takes place across an a i r space.
The action
"The action of spores on the sulphur i s prim arily a reducing one.
Hydrogen sulphide i s extremely toxic to fungous spores, much more so than
any other sulphur acid.
The order of s e n sitiv ity o f the spores of
d ifferent species to sulphur and to hydrogen sulphide is id en tical and
the sensitive spores produce morfe than one toxic u n it o f hydrogen sulphide,
while the re sis ta n t ones produce considerably le ss than one toxic u n it.
F inally, compounds such as glutathione, which contain the -S H group,
readily react with sulphur producing hydrogen sulphide a t ordinary
temperatures, and such compounds have been shown to e x ist in fungous spores.
"The following in terp retatio n of these facts as rela ted to the
fungicidal action of sulphur i s offered.
Sulphur in the v ic in ity of fungous
spores, by reason of i t s vapor pressure, gives o ff sulphur vapor which
diffuses into the spores.
Here reduction takes place within the spores
with hydrogen sulphide as a fin a l product.
Die reaction i s enzymatic in
nature and is probably concerned with —S H compounds.
The toxic product,
hydrogen sulphide, being produced in intim ate contact with the liv in g c e ll
is able to exert i t s maximum e ffe c t.
I t is not believed th at the hydrogen
sulphide produced from the leaves in the open effects the spores, nor th at
the hydrogen sulphide produced by one spore has much e ffe c t on another
spore a t a distance.
Each individual spore, therefore, by reason o f i t s
a b ility to reduce sulphur to hydrogen sulphide, i s thus instrumental in
bringing about i t s own death."
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BXPEBIliIENTAL METHODS AND EESULTS
The Production of Hydrogen SuliaMde ty SnlBhur Treated Insects
In order to determine the role of hydrogen sulphide in the to x ic ity
of sulphur to in sects, the f i r s t step necessary was to te s t th e ir a b ility
to produce hydrogen sulphide when treated with sulphur.
wsus as follows:
The method used
The samples of insects o r m aterial to be tested were
placed in sim ilar v ia ls, a thorou^x dusting of flowers of sulphur applied,
and the v ia ls tl^ a tly stoppered with corks from which s trip s of f i l t e r
paper moistened with saturated lead acetate solution were suspended as
Indicators.
The v ia ls were then placed under the desired conditions of
temperature fo r the duration of the experiment, observations being made
at frequent in te rv a ls.
The production of hydrogen sulphide was shown by
a black p re c ip ita te of lead sulphide on the f i l t e r paper.
These te s ts
were run with liv e in sects, garden slugs, fie ld spiders, clover m ites,
in sects with in te stin a l tr a c t removed, the in te s tin a l tra c t i t s e l f , ground
up Insect tissu e , etc.
The re su lts were p o sitiv e in every case, and
varied with d ifferen t species.
Controls with these same m aterials, under
the same conditions except sulphur, also gave positive re su lts except with
most of the In te stin a l tr a c ts and ten t c a te rp illa r larvae.
In case of
the controls, as may be seen in Table I , the production was much slower
than with treated m aterial.
No hydrogen sulphide was produced in controls
with sulphur alone or as a paste.
Most of th e species used are not ^
affected by sulphur and Colorado potato beetles seemed to 'farw -b etter in J
sulphur than in the untreated checks, yet they produced hydrogen sulphide
re a d ily ..
A possible explanation of th is fact Is the adverse action
o f the hydrogen sulphide produced upon the putrefying b ac te ria associated
with the in sect.
The production o f hydrogen sulphide by untreated m aterial
resu lts from the decomposition of proteins by putrefying b acteria.
Almy (1925) sta te s that hydrogen sulphide is one of the end products
resulting from the action of many v a rie tie s o f b acteria on organic m aterial
containing p ro tein .
This author cred its Bettger1 as sta tin g that hydrogen
sulphide i s , no doubt, one of the f i r s t substances which i s s p lit o ff from t
the protein molecule during putrefaction.
A quantitative determination of the amount of hydrogen sulphide
produced by insects trea ted w ith sulphur i s , by any method suggested
heretofore, impossible.
The amount produced i s small, and as i t I s
produced in the tissu e where i t is combined with end absorbed by various
substances, on the cu ticu la, and walls of the container, any quantitative
estimation would be u n reliab le.
A method decided upon which would seem to give a somewhat
comparative quantitative estimation i s given below.
The blackening of.
the f i l t e r paper in d icato r, as hydrogen sulphide was produced, always began
a t the bottom and proceeded gradually upward, so approximate standards
were made and applied to the amount of the paper blackened a f te r a ce rtain
time.
From the application of these standards, the re s u lts shown in
Table I were obtained.
5 o r G te s ts .
The re su lts in Table I are averages o f from I to
I f no blackening occurred, 0 o r no te s t was recorded; from
I - Bettger. Jour. Biol. Chem. 2:71, 1906- 7.
the slig h te st darkening a t the tip to 1/3 black was recorded as X or
l i g i t te s t; 1/3 to 2/3 black as medi-um te s t or XX; 2/3 to fu lly covered
as XXX or heavy te s t, and very black as XXXX o r very heavy te s t.
I t was thougit th a t by in jectin g lead acetate solution into the
body tissu e of c le a r skinned species, and tre a tin g with sulphur, th at
possibly the p recip itatio n of black lead sulphide would show the progress
of hydrogen sulphide production in the body.
Large larvae of white grubs and round-headed wood borers were
selected fo r the te s t.
The lead acetate solution, however, k ille d the
larvae soon a fte r in jectio n and the experiment was not successful*
Some
of the larvae were placed in v ia ls, treated with flowers of sulphur, and
kept overnigit.
Upon in jectio n the following morning, some p re c ip ita te
was shown near the point of in jectio n .
This p re c ip ita te was not very
noticeable, and was not shown over a l l p a rts of the body.
These larvae do
not contain much firm tissu e , and most of the solution escaped througi
the small hole made by the hypodermic needle as soon as i t was removed.
This probably, in p a rt, accounts fo r fa ilu re o f other p a rts of the body
to show a p rec ip ita te .
A white grub larv ae, injected with lead acetate and placed in
a concentration of hydrogen sulphide of 5 milligrams p er l i t e r , immediately
turned black a l l over.
These species are not very much affected by sulphur, and do not
show the production o f hydrogen sulphide very readily.
The hydrogen
sulphide is produced in the tissu e and i t s production i s very slow,
la y
combination with and adsorption b% substances in the tissu e takes place
as rapidly as i t is produced u n til such combining and adsorbing capacities
TAMM I .
SEOtrai TM PfflSJOSIOF Ol1 EYDSOGBE S U IfE ia IT SUEEBUB TSSiOD IESSOTS
Material
I h r.
Colorado potato "beetle, Leotlnotarsa decemllnea.taf Adults 0*
e
N
R
"(le ss i n t . tra c t) O
w
M
H
"(D itto ground up) O
N
H
R
" ( in t. tra c t)
O
N
R
R
" lanrae O
s
H
R
" (le s s i n t . tra c t
ground up)
O
" (in te s tin a l tr a c t)0
Garden slugs, Agrlollmax agrestes
O
R
H
R
H
(ground up)
O
Grasshoppers, Melanonlus M r ltta tu s
O
"
"
(in te s tin a l tra c t)
O
M
(le ss in t. tra c t)
O
SI
( " "
",ground up O
Imported cabbage worn, P le ris ranae. larvae
O
"
*
"
" (less in t, tra c t)
O
"
"
"
"
" ( in t. tra c t)
"
"
"
"
" (excreta)
O
Mites, clover, Schlsotetraarchua nratensis
O
Sugar beet webworm, Loxostege a t l c t l c a l l s .. larvae
x
Tabanid larvae
O
Tent c a te rp illa r, Malacosoma spp., larvae
O
"
".
"
" (ground up)
O
Trlbolium confuslua. adults
O
Sulphur paste, d ifferen t d ilu tio n s as check
O
Treatment
_ Sulphur
Ho Snlphnr
4 hre 2Mhrs IZhxn. I h r. 4 h r a. 24 hra. 72 h r*
xx
XXXX
O
O
x
XXX
x
XXX
XXXX
O
O
X
XXX
•x
XX
XXXX
O
O
X
XXX
X
XX
XXX
O
O
O
O
X
XX
XXX
O
O
X
XXX
X
X
X
X
O
X
X
X
O
O
X
X
X
X
O
O
O
O
XX
XXX
XX
XXX
X
XXX
XXX
XXX
XXX
XX
XXX
XXX
XXX
XX
X
X
X
XXXX
XXX
XXX
XXXX
XX
XXX
XXX
XXX
XXX
XXX
XXX
XXX
XXX
XXX
XX
XX
O
*- O no te s t, x lig h t te s t, xx medium te s t, xxx heavy te s t, xxxx very heavy te s t.
O
O
O
O
O
O
e
O
O
O
O
O
O
O
O
O
O
O
O
O
O
O
O
O
O
O
O
O
O
O
O
O
O
O
X
XX
O
O
O
X
O
O
x
X
X
X
XXX
XXX
X
O
XX
XX
XX
X
XX
XX
X
XX
XX
X
XX
O
O
O
O
O
O
r
-1 9 -
are exhausted.
T htll there is hydrogen sulphide present in excess of
th at combined or adsorbed, no lead sulphide w ill be p recip itated upon
injection with lead acetate.
In view o f these
facts i t i s not surprising
th at no great amount o f blackening was shown, in case of sulphured larvae.
The S ffect o f Temperature on the Reaction
The ra te of hydrogen sulphide production gradually Increased
from room temperature, about 22% up to 37eC*» which was the highest
temperature used.
The increase was not as great as expected, which can be
explained by the reasons given for making quantitative determination
impossible.
The E ffect o f Hydrogen Ion Concentration on the Beactlon
Tests were run on yeast in solutions of pH value from I to 12.
t h fit
The re su lts were so nearly the SamexTzatil i t seems that the hydrogen ion
concentration does not e ffe c t the reaction within these lim its.
These
re su lts are in agreement with those of McCallan and Wllcoxon (1931) on
th is point, interpreted by than as fu rth er evidence in favor of the
conclusion th at the production of hydrogen sulphide goes on inside the
organism.
Ho sulphur sen sitiv e species were available in su fficien t
quantities to be used fo r these te s ts .
to run one tes^bn a small scale.
Only enough mites were secured
Species Which are controlled by sulphur
might produce hydrogen sulphide much more read ily than most of the species
used.
-2 0 -
HyArogen Sulphide Prodactlon 'by Lime-Stdphur
In a series of te s ts , under the conditions outlined fo r the
experiments with sulphur, hydrogen sulphide was evolved readily from
dry lime-sulphur, alone, as removed from commercial containers.
Solutions
of th is m aterial, made ty adding 2 teaspoonsfu l to a p in t of water,
produced hydrogen sulphide in appreciable q u an tities, when applied to
insects o r alone.
The residue from th is solution, a f te r evaporation to
dryness in the open a i r , also , produced th is gas alone o r when added to
insects.
The controls, with the m aterial alone, produced hydrogen
sulphide in each case hut not as rapidly or in as great qu an tities as
when applied to in sects.
Colorado potato beetles were used in these te s ts .
The Toxicity o f Sulphur to Some Species
Realizing th at the species available fo r experiment were not
capable of being controlled by sulphur, i t was desirable to learn whether
they were a t a l l affected by th is substance.
The methods used and re su lts
obtained follow.
Four male and U female grasshoppers, (Kelanoulus b lv itta tu s) were
placed in a large v ia l, thorou^aly dusted with flowers o f iulphur, and
tig h tly stoppered.
An equal number of as near the same size and a c tiv ity
as possible, were placed under exactly the same conditions, except sulphur
■tZ*. *-t
Aas checks. A fter 10 hours a l l specimens, both treated and untreated, had
ceased a l l a c tiv ity and were apparently dead.
They were then taken out
of the v ia ls and placed separately in screen topped containers fo r
observation.
Vlthin 5 minutes two of each had recovered su ffic ie n tly to
-2 1 -
Junrp against the tops of the containers.
At the end of 12 hours, a l l
of the untreated Individuals had apparently recovered completely.
At
th is time 2 of the treated females were apparently dead, and 2 had only
recovered enou^i to hold on to the finger when picked up.
females never recovered.
The weakened
Under the conditions of the experiment, sulphur
Is apparently somewhat toxic to th is species.
Colorado potato beetles given the same treatment seemed to be
slig h tly weakened, but a f te r 4 hours In the open a i r recovery was complete.
Subjected to the same conditions for 24 hours, Trlbollum confusum
showed no i l l e ffe c ts.
The same was true fo r Sltouhllus oryzae.
The Relationship of the Glutathione Content of a Suedes
to the A b ility o f that Suedes to Produce
Hydrogen Suluhlde from Suluhur.
The a b ility of glutathione, a trlp ep tld e, to reduce sulphur
to hydrogen sulphide i s well established.
This trlp ep tld e, of glycine,
glutamic acid and cysteine, has been studied quite exhaustively by
Hopkins, Tunnlcliffe (1925), Mason (1931), Meldrum and Dixon (1930),
and others.
This substance is widely d istrib u ted In liv in g organisms.
To determine the correlation between the glutathione content of various
species and th e ir a b ility to produce hydrogen sulphide when treated with
sulphur, might .throw some lig h t on the nature of the reaction.
Some e ffo rt
was directed along th is lin e . . The method of Fink ( 1927) with minor
modifications was used in the determinations.
The procedure is as follows;
Approximately 10 grams of Insects or m aterial used was weighed
to four decimal places, cut up, placed In a glass mortar with a l i t t l e
sand and a few cc. of a 10 percent trich lo ra cetic acid solution, and
-2 2 -
gro-and np thoron^ily,
This mixture was then placed In a fla sk ,Aenough
additional acid solution added to cover the m aterial.
then tig h tly stoppered and l e f t fo r about 10 hours.
The flask was
The m aterial was
filte re d , the residue placed back In the flask , go re. acid added, and
l e f t fo r 5 hours when I t was f ilte re d again.
and the following determination run.
The f iltr a t e s were combined
Five cc. of the f i l t r a t e was drawn
out, placed in a beaker and titr a te d with approximately 0.0 I U Iodine
solution, using starch solution fo r an Indicator.
The stock Iodine
solution was standardized before each titr a tio n by titr a tin g with
standard sodium thiosulphate solution.
The c h itln content was estimated
by removing the tissu e and other foreign m aterial from weighted samples
with 10 percent sodium hydroxide solution.
The insects were cut open,
placed in the solution, l e f t overnight, removed,washed thorou^ily, dried
In a sulphuric acid desiccator and the remaining m aterial calculated as
the c h itln content.
The percentage of water was determined by
dehydration throu^i a se rie s of from 6 0 to 9 0 percent alcohol and a
sulphuric acid desiccator.
The amount of glutathione contained in milligrams per gram of
tissu e was calculated on the b asis, as used by Fink ( 1927), that I cc. of
0.0 I B iodine is equivalent to 2.5 milligrams of glutathione.
re su lts are shown in Table I I .
of from I to U te s ts .
The
Each re su lt in th is table Is the average
-2 3 -
TABLB I I .
GLTJTATHIOIIB COHTBHT OS* SOME IHSBCTS, SLUGS, ETC.
Material Tested
Average, mg. per gm.
Leutlnotarsa decemllneata, adults
*
*
larvae
Melanoulus b lv ltta tu s
Slugs. Agriollmax agrestes
Flelschmans yeast
Corn, Sea mays
Lettucer Lactuca satlva
Potatoes, Sglemzn tuberosum, tubers
L ilac. Syringa vulgaris
2.U2
1.90
1.10
1.46
6. S3
1.13
1.76
0.47
0.82
I t Is realized th at these re su lts are not altogether accurate
fo r several reasons, some o f shich follow.
F irs t, the experimental
procedure was not such as to produce accurate re su lts.
Second, the
d iffic u lty In determining exactly the end point in the titr a tio n makes the
re su lts questionable.
Third, the lik e ly presence \ o f -S H groups besides
those in uhdlssociated glutathione would make the method unreliable.
However, as sim ilar procedure was followed in a l l the determinations, i t
is thou^rt that the re su lts might answer fo r comparative purposes.
From the re su lts obtained, i t appears th at there might be some
p o sitiv e correlatio n between the glutathione content of a species and i t s
a b ility to produce hydrogen sulphide from sulphur.
However, the lim ited
number o f determinations and the method used do not permit d efin ite
conclusions.
The Effec t of Sulrhur on the Catalase A ctivity of
Melanoulus b lv ltta tu s .
Shafer (1915) found th a t insecticides affect to a g reater o r
Ieeaer extent the enzymatic balance between the oridaeee, reductasef
and catalaaeS
As equipment was available with which the effect on
catalase could be measured, the following te s ts were made.
Four male
M. b lv itta tu s were placed in v ia ls and thoroughly dusted with flowers of
sulphur.
Four males of the same species, as near the same size as the
treated ones as possible, were placed in sim ilar v ia ls under the same
conditions except dusting.
All v ia ls were tig h tly stoppered for about
10 hours, when the stoppers were slig h tly loosened to prevent suffocation.
A fter 24 hours, each grasshopper was weighed, ground fo r 5 minutes in a
mortar, and trea ted with excess hydrogen peroxide in a gas measuring device.
The basis of comparison was the average amount of free oxygen lib e ra ted
p er second from I gram of pulp from tre a te d and untreated individuals.
The evolved gas was measured in a gas b u rette over water a t 20*0.
The
average amount of oxygen lib e ra te d per second from I gram of pulp of
untreated
b lv itta tu s was 2.7 cc. compared with 2.31 cc. per second
per gram fo r sulphur trea ted individuals.
These re su lts Indicate that the a c tiv ity of the catalase in the
insect#s body may be somewhat inhibited by treatment with sulphur.
However,
th is Inhibition i s probably not enough to upset the enzymatic balance to
any Important extent.
The Toxicity o f Hydrogen Suluhlde to Insects
The toxic component o r property of sulphur must be associated
with th a t element under conditions as found when i t is applied in the fie ld .
The component or property to which the g reatest value i s a ttrib u te d , other
-2 5 -
thlngs being equal, must prove the most toxic vhen te sted under controlled
conditions.
To determine the statu s of hydrogen sulphide as a toxic agent
In the action of sulphur, to x ic ity te s ts were ran with th at gas and the
"median le th a l concentration* calculated.
Strand (1930) showed that as a basis fo r comparison, the
"median le th a l concentration o r dosage" "Was by fa r the best method of
expressing toxic concentrations. He also showed th at i f the concentration
Is varied, time and temperature should be held constant.
Tlve hours was
selected as the most convenient time of exposure. By "median le th a l
concentration" Is meant the concentration of the substance required to
k i l l 50 percent o f the organisms in consideration under the conditions of
the experiment.
In th is case, i t means the concentration in milligrams
p er l i t e r necessary to M i l 50 percent of Trlbollum cpnfusum a t 250C. -under
5 hours exposure.
As past h isto ry very lik e ly affects the physiology o f
insects with respect to adsorption and diffusion of gases, and other
factors rela ted to to x ic ity , i t is necessary that the specimens used in
comparative te s ts be reared under sim ilar conditions.
The insects used
in th is case were taken from a culture of confused flo u r beetles which
were kept a t approximately 2$%.
The experimental procedure was as follows.
Bolting cloth
cages containing about 30 T. confusum were suspended about 2 or 3 inches
from the bottom of. 6.U l i t e r Brlenmeyer flask s, which were then stoppered
with glass stoppers. Bach of these stoppers was: constructed with two
glass tubes throu^i i t , which were provided with stopcocks.
A p a rtia l
vacuum was created in each flask by drawing out a ll the a i r one could
by mouth.
One of the stopcocks was attached to a gas b u rette.
The
-2 6 -
d e s ir e d c o n c e n tr a tio n o f hy d ro g en e u lp h id e was drawn I n to th e f l a s k , th e n
th e o t h e r s to p c o c k opened w hich a llo w e d a i r to ru s h I n u n t i l th e p r e s s u r e
in s id e e q u a lle d t h a t o f th e stmos p h e re .
a la r g e te m p e ra tu re c a b in e t a t 2 $ % .
The f l a s k s were th e n p la c e d i n
At th e end o f 5 h o u rs th e c ag e s
w ere removed, an d th e number o f dead and l i v e i n s e c t s re c o rd e d .
c o u n t was made J 2 h o u rs a f t e r rem oval.
A fin a l
The c o n c e n tr a tio n i n m illig ra m s
p e r l i t e r an d th e p e r c e n t m o r t a l i t y was c a l c u l a t e d f o r each t e s t .
two t e s t s w ere ru n w ith 2 c a g e s o r 60 i n s e c t s p e r t e s t .
F o r ty -
The p e rc e n ta g e
m o r t a l i t y i n th e checks was so sm all t h a t i t was n e g le c te d .
From th e
r e s u l t s o b ta in e d , th e t o x i c i t y c u rv e , shown i n F ig u re I , was made by
p l o t t i n g c o n c e n tr a tio n i n m illig ra m s p e r l i t e r a g a in s t p e rc e n ta g e m o r t a l i t y
and draw ing th e b e s t c u rv e th ro u g h th e p o i n t s .
The "m edian l e t h a l
c o n c e n tr a tio n " o f hydrogen s u lp h id e f o r T. confustra a s c a l c u l a t e d from
t h i s c u rv e was v e ry n e a r to 3.&> mg. p e r l i t e r .
The minimum l e t h a l
c o n c e n tr a tio n was ab o u t 6.8 mg. p e r l i t e r o r 0 . 5 p e rc e n t b y volume.
T h is i s 1.1 mg. p e r l i t e r lo w e r th a n S tr a n d (1 9 3 0 ) o b ta in e d
f o r c h l o r o p i c r i n and 4 .3 3 mS- Per l i t e r lo w e r th a n h e o b ta in e d f o r ammonia.
I t i s a l i t t l e o v e r 9 tim e s a s to x ic a s e th y le n e d ic h lo r id e , 16.9 tim e s a s
to x ic a s carb o n d is u lp h id e and l / 6 a s to x ic a s h y d ro c y a n ic a c id g a s,
when com pared w ith th e r e s u l t s o f t h a t same a u th o r.
A few t e s t s w ith th e r i c e w e e v il, S ito u h llu s o rv z a e . show th e
m edian l e t h a l c o n c e n tr a tio n o f hydrogen s u lp h id e f o r t h i s s p e c ie s to be
n e a r 6.8 mg. p e r l i t e r .
The d i f f e r e n t s p e c ie s , th e r e f o r e , v a ry in t h e i r
r e s i s t a n c e to t h i s g a s .
Ho c o n s id e r a tio n was made f o r a d s o r p tio n o r d i f f u s i o n .
u s e d was a com m ercial p ro d u c t and c o n s id e re d a s 100 p e r c e n t p u r e .'
The g a s
The
-27-
c o n c e n tr a tio n was, t h e r e f o r e , p ro b a b ly , low er th an t h a t c a lc u la te d .
T e s ts show b o th T. c o n fu gu-n and S. o ry zae to be v e ry s l i g h t l y
i f a t a l l a f f e c t e d by s u lp h u r. I t i s v e ry l i k e l y , t h e r e f o r e , t h a t t o x i c i t y
t e s t s w ith hydrogen su lp h id e on e x tre m ely s u lp h u r s e n s i t i v e s p e c ie s , as
r e d s p id e r s o r c o tto n fle a h o p p e r s , would show t o t a l m o r t a lit y a t much low er
c o n c e n tr a tio n s th a n n e c e s s a ry f o r th e s p e c ie s t e s t e d . I t i s n o t a t a l l
d i f f i c u l t to im agine such a c o n c e n tr a tio n in o r r i g h t around an i n s e c t 's
body when i n c o n ta c t w ith s u lp h u r, even in th e open a i r .
U n fo rtu n a te ly no su lp h u r s e n s i t i v e s p e c ie s were a v a il a b le i n
s u f f i c i e n t q u a n t i t i e s f o r t o x i c i t y t e s t s . Such b e in g th e c a s e , i t was
im p o s sib le to c o r r e l a t e , any f u r t h e r , s u lp h u r s e n s i t i v i t y w ith th e t o x i c i t y
o f hydrogen s u lp h id e .
■
too
o o
3
80
0Z f
3
O
r
cf
5
>o
I o
o
X
k
*
O
O
.
:
I
9
<
= O
'
90
8
'
/ 7 . L i Z.
3 . 6 Mgt 'L.
I
0S
■
'
C O N C N ., M g l L I T E J S
F ig u re I . T o x ic ity Curve f o r Hydrogen S u lp h id e From Which th e Median
L e th a l C o n c e n tra tio n f o r th e Confused F lo u r B e e tle Was
C a lc u la te d .
-2 8 -
t
DISCUSSION AND CONCLUSIONS
Some facts to Ue considered in drawing a conclusion as to the
toxic action of sulphur to insects are as follows.
Some detrimental effect of lime-sulphur solution i s possibly
due to the imprisonment of the young under the wax covering of the mother
scale.
Shafer (1911) showed the freshly secreted margins o f these scales
to he softened, in some cases, causing them to adhere to the baric.
Cooley (1910), however, shows the real effect of lime-sulphur
to Ue exhibited on the young scale in sects, immediately a f te r hatching,
often a month o r six weeks a f te r application of the spray.
Studies on
the chemical composition o f lime-sulphur show the tendency fo r free sulphur
to be deposited as the fin a l product.
By the time the young scales hatch,
p ra c tic a lly a l l the sulphur in the dormant o r delayed dormant spray has
been deposited as the free element.
The consensus of opinion as to the action of I Ime-sulphur
points to some property o f the element i t s e l f .
McGregor ( I 927), working with in se cts, and McCallan and Wilcoxon
(1931)»
and others, working with fungi, have shown th at the action can
take place across a space.
The fact that the toxic action can be exerted across a space
leads to the conclusion th at sulphur enters the body of the insect as
gaseous o r sublimed sulphur.
McCallan and Wilcoxon (1931) showed sulphur se n sitiv ity of eight
species of fungi to correspond p erfectly with the to x ic ity of hydrogen
-2 9 -
sulphide to the same species.
They concluded from th e ir re su lts that
each fungus spore hrings about i t s own death when trea ted with sulphur
by reducing the element to the extremely toxic hydrogen sulphide.
P articu late sulphur i s usually thought of as a rath er in e rt
substance, but the vapor has greatly Increased surface and other physical
differences making the p o ssib ility of a much more active m aterial.
As to the toxic action of sulphur, there is no doubt that death
in most cases is brought about by the portion that enters the in s e c t's
body in the gaseous sta te , and evidence favors the conclusion that the
fa ta l re su lt i s caused by the hydrogen sulphide i&lch i s produced from
i t s reduction inside the body.
-3 0 -
m m m
A somewhat extensive review of viewpoints dealing with the toxic
action of sulphur and lime-sulphur solution, presented in the readily
available lite r a tu r e , is Included.
The leading reports on chemical Investigations of lime-sulphur
solution are reviewed.
The readiness with which sulphur, i s reduced to hydrogen sulphide
by liv in g insects is shown.
Experiments are described which show th is gas to be highly toxic
to sulphur re sista n t species.
The inference Ie made th a t i t would lik e ly
prove much more toxic to sulphur sensitive species.
Results of other workers show th at the toxic action can be
exerted across a space.
I t is concluded that the portion of the sulphur which does
most of the damage to in sects enters the body as gaseous sulphur.
Whether i t produces i t s toxic action upon insects in th is form or is
reduced to hydrogen sulphide which brings death, can only be inferred
from in d irec t evidence.
-3 1 -
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IlISWi