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Lysine requirements of larvae of Oryzaephilus surinamensis (L.) (Coleoptera, Silvanidae)
J. Ins. Physiol., 1961, Vol. 6, pp, 122
to 125. Pergamon Press Ltd., London. Printed in Great Britain
LYSINE REQUIREMENTS OF LARVAE OF ORYZAE~H~L~~ SURINAMENSIS (L.) (COLEOPTERA, SILVANIDAE)* G. R. F. DAVIS Entomoiogy
Section, Research Station, Research Branch, Canada Agriculture, Saskatoon, Saskatchewan, Canada (Received 15 November
1960)
Abstract-Larvae of the saw-toothed grain beetle, Oryxaephilus surinamtxsis (L.), survived in greater numbers and developed quicker as the concentration of L-lysine in the diet increased from 0 to I.37 per cent. At higher concentrations the number surviving and the rate of growth decreased. Of related compounds tested, pipecolic acid, cr-aminoadipic acid, or a mixture of or-ketoglutaric acid and ammonium acetate partiaily supported growth; glutaric acid, ar-ketoglutaric acid, ammonium acetate, dipheny~hydantoin, or hydantoin did not.
INTRODUCTION LYSINE
is considered an essential amino acid for most organisms. However, it can be formed from aspartic acid by Esche-richia cdi (BOLTON et aZ., 1953) ; is essential for Streptococcus faecalis (6057) only under certain conditions (MCCLURE et al., 1954); and is synthesized by ~~tte~~a g~rn~~i~~ L., but not rapidly enough to satisfy the requirements of the organism (GORDON, 1959). Extensive reviews on lysine requirements and metabolism have been published by WORK (19%) and MEISTER (1957). DAVIS (1956) indicated that Oryzaephilus surinamensis (L.) could possibly dispense with lysine in the diet, At the time, because of the nature of the diet, absolute requirements of this organism for amino acids could not be determined. A chemically defined diet in which lysine can be a limiting amino acid has since been formulated. This is a report on the quantitative dietary requirements of larvae of 0. surinam~~~ for lysine. The effects of related compounds as replacements for lysine were also studied. MATERIALS
AND METHODS
The
components of the chemically defined diet were the same as those used previously (DAVIS, 1959) except that L-alanine and L-proline were included at the O-07 per cent level. VVith the same method as in earlier work (DAVIS, 1959), the effects of L-lysine were tested at twelve concentrations from 0 to 2.39 per cent. All constituents of these diets were obtained from Nutritional Biochemicals Corporation, Cleveland, Ohio. * Contribution
No. 65, Canada Department
of Agriculture
122
Research Station, Saskatoon, Sask.
LYSINE REQUIREMENTS OF LAEVAEOF ORYZAEPHILUS
SURINAMENSL!3
(L.)
123
The following related compounds were substituted for lysine in the diet: Dr..-pipecolic acid hydrochloride* at two concentrations; DL-~-a~noadipic acid* at four; and glutaric acid,$ a-ketoglutaric acidx (alone and with ammonium acetates), ammonium acetate, diphenylhydantoin,” and hydantoinf at one concentration each. RESULTS
AND DISCUSSION
Table 1 shows that as the percentage of lysine in the diet of the saw-toothed grain beetle increases, both survival and rate of development increase until a m~mum is reached at a.concentration of 1.37 per cent. As the concentra~on of iysine is further increased, survival and rate of development decrease rapidly. TABLE ~-THE NUMBER OF INDIVIDUALSPUPATING AND EMERGING AS ADULTS, AND THE AVERAGETIMESTo PUPATIONAND EMERGENCE WHEN 100 LARVAEOF Oryzaephilus sutinamensis WERE REAREDINDIVIDUALLYON RIETS CONTAININGVARIOUSQUANTITIESOF L-LYSINE Percentage L-lysine in diet ~-
Number pupating
::: 0.02 0.04 0.08 0.17 0.34 068 1.37 1‘71 2*05 2.39 LSD. at 5% level
; It 27 :: 70 74 27 :: 29
Average time (hr)
1764 1393 * 1353k 1227f 13.59 + 943 * 822 + 743+ 707* 1048+ 1002+ 1291 + -
144* 97 77 100 33 29 17 Ifs? 21 48 76
Number emerging ii ii: 20 :: t: 19 f 22
Average time (hr) 1884 1532 + 141* 1382+115 1336+ 84 1501+112 1066+ 38 968k 33 8632 19 788+ 1st 1169f 43 1122rt 54 1.5112 97 -
* Standard error. f Significantly less than other values at the 5 per cent level of confidence.
gos~~~ilu (Saund.) (V~DE~ANT, 1958); Apis rne~~~c~ L. (DE 1953) ; Aedes ffeg~p~~(L.) (GOLBERG and DE MEILLON, 1948 ; SINGH and BROWN, 1957) ; Calliphoru erythrocephalu (Meig.) (&DEE, 1954) ; Phormiu regina (Meig.) (MCGINNIS et al., 1956; CHELDELIN and NEWBURGH, 1959); Pseudosarcophaga a$%& (Fall.) (HOUSE, 1954); ‘Attugenus sp.’ (MOORE, 1946); Tribolium confusum DIN. (LEMONDE and BERNARD, 1951; FRAENKEL and PRINTY, 1954) ; and Trogoderma gvanarium Everts (PANT et al., 1958) require lysine for growth and development. The present work shows a definite requirement for lysine in the diet of the saw-toothed grain beetle. The results of earlier work (DAVIS, 1956) which indicated that 0. ~7~na~~ could dispense with lysine in the diet may Pec~~n~~~u
GROOT,
* Mann Research Laboratories, New York, N.Y. t Eastman Organic Chemicals, Rochester, N.Y. $ Nutritional Biochemicals Corporation, Cleveland, Ohio. 5 Fisher Scientific Company, Fairlawn, N. J.
G. R. F. DAVIS
124
have occurred because of impurities in the amino acid constituents (GORDON, 1959) or because of the presence of micro-organisms in the diet capable of converting aspartic acid to lysine (BOLTON et al., 1953). Table 2 indicates that larvae of 0. surinamer~+ on a lysine deficient diet may be able to utilize pipecolic acid, or-aminoadipic acid, or a mixture of or-ketoglutaric acid and ammonium acetate. These compounds either increased the rate of development or the survival of larvae reared without lysine. Glutaric acid, ol-ketoglutaric acid, ammonium acetate, diphenylhydantoin, or hydantoin did not. TABLE ~-THE NUMBEROF INDIVIDUALS PUPATINGAND EMERGINGAS ADULTS,AND THE AVERAGE TIMESTO PUPATIONANDEMERGENCE WHEN100 LARVAE OF Oryzaephilus surinamensis WRRE
REARED
INDIVIDUALLY
ON
DIRTS
COMPOUNDS
Lysine replaced by (%)
.
Number pupating
IN
-
_-
WHICH
VARIOUS
REPLACED
L-LYSINE
CONCENTRATIONS
OF
RELATED
-
Average time (hr)
Number
--
,emerging
Average time (hr)
DL-Pipecolic acid HCI
I.24 2.48
nr_-ar-Aminoadipic acid 1.21 2.42 3.63 4.84 Glutaric acid
l-37 c+Ketoglutaric acid 1.37 cY-Ketoglutaric acid + ammonium acetate 0.46 + 0.92 Ammonium acetate 1.37 Diphenylhydantoin 2.06 Hydantoin 0.94
2 0
972 + 34* 1156 &46 1150+53 -
1; :
2
1128 f26* -
7 10 -
1353 +45 1272 k47 -
-
0
-
0
-
18
11
1583 +93
1
1332
0
-
1740f125
1
0
-
1428
-
-
-
-
-
* Standard error.
The proposed pathway of lysine metabolism in the rat is as follows: lysine -+cu-aminoadipate+glutarate+ c+ketoglutarate and acetate (ROTHSTEIN and between lysine and or-aminoMILLER, 1954). MEISTER (1957) inserts pipecolate adipate. The results of the foregoing experiments suggest that this pathway is only partly operative in larvae of 0. Suriname&s, because the effective compounds substituted for lysine did not permit either as rapid development or as great survival as lysine itself. For the same reasons, the results indicate that these compounds, in the concentrations used, could not substitute entirely for lysine in the diet of 0. surinamensis. Acknowledgement-The acknowledged.
technical
assistance
of
Mr.
F.
T.
EVES is
gratefully
LYSINEREQUIRsMENTs OF LAnVAEOF ORYZAEPHILUS SLJRIiVAMENS1.S (L.)
12.5
REFE~NC~S BOLTONE. T., BRITTENR., and COWIE D. B. (1953) Synthesis of the aspartic and glutamic families of ammo acids by Escherichia coli. Science 17, 465. CHBLDELINV. H. and NEWBURGH R. W. (1959) Nutritional studies on the blowfly. In: Axenic culture of intervertebrate metazoa: a goal. Ann. N. Y. Acad. Sei. 77, 373-383. DAVIS G. R. F. (1956) Amino acid requirements of Ory,zuephiZus surirramensis (L.) (Coleoptera : Silvanidae) for pupation. Canad. J. 2001. 34, 82-85. DAVISG. R. F. (1959) Alanine and proline in the diet of larvae of Oryzaephilus su~~rn~~ (L.) (Coleoptera : Silvanidae). Arm. cnt. Sot. Ames. 52, 164-167. DE GROOTA. P. (1953) Protein and amino acid requirements of the honeybee (A&s mellijka L.). Physiol. camp. 3, l-89. FRAENKELG. and PRINTY G. E. (1954) The amino acid requirements of the confused flour beetle, Tribolium confusum Duval. B&Z. Bull., Woo& Hole 106, 149-157, GOLBERG L. and DE MEILLON B. (1948) The nutrition of the larvae of Aedes aegypti Linnaeus-IV. Protein and amino acid requirements. Bio&em. J. 43, 379-387. GORDONH. T. (1959) Minimal nutritional requirements of the German roach, Blattella germanica L. In: Axenic culture of invertebrate metazoa: a goal. Ann. N. Y. Acad. Sci. 77, 290-351. HOUSE H. L. (1954) Nutritional studies with Pseudasurcophaga a#+& (Fall.), a dipterous parasite of the spruce budworm, Choristonewu fzm$erana (Clem.)-HI. Effects of nineteen amino acids on growth. Canad, J. Zool. 32, 351-357. LEMQNDEA. and BERN- R. (1951) Nutrition des larves de Tribolium confusum DuvalII. Importance des acides amines. Car&-J. 2001. 29,80-83. MCCLURE L. E., NEUMANR. E., and MCCOY T. A. (1954) Amino acid metabolic studiesVI. Aspartic acid-lysine interrelations in Streptococcus faecalis (6057). Arch. Biochem. Bio@hysr. 53, 50-55. MCGINNIS A. J., NEW~URGHR. W., and CHELDELINV. H. (1956) Nutritional studies on the blowfly, F%orm& reg& (Meig.). g. N&r. S8, 309-324. MKIS-IERA. (1957) Bhhhtry of the Amino Acids. Academic Press, New York. MOOREW. (1946) Nutrition of Attagenus (I) sp. Ann. ent. Sot. Amer. 39, 513-521. PANT N. C., NAYARJ. K., and GUPTA P. (1958) On the significance of amino acids in the larval development of Khapra-beetle, Trogoderma granarium Everts. (Coleoptera : Dermestidae). E~&etttia 14, 176-178. ROTHSTEINM. and MILLER L. L. (1954) The metabolism of r&sine-6-C14. J. biol. Cha. 206, 243-253. SEDEE J. W. (1954) Qualitative amino acid requirements of larvae of Calliphora erythro.. cephala (Meigen). Acta n&l. phawn. physibl. 3, 262-269. SINGH K. R. P. and BROU~NW. A. (1957) Nutritional requirements of Aedes aegypti L. J. Ins. Physiol. 1, 199-220. VANDEIUANT E. A. (1958) The amino acid requirements of the pink bollworm. g. econ. Ent. 51, 309-311. WORK E. (1955) Some comparative aspects of lysine metabolism. A Sympoti~m on Amino Acid Met&o&m (Ed. by W. E. MCELROY and B. GLASS), pp. 462492. The Johns Hopkins Press, Baltimore.
to 125. Pergamon Press Ltd., London. Printed in Great Britain
LYSINE REQUIREMENTS OF LARVAE OF ORYZAE~H~L~~ SURINAMENSIS (L.) (COLEOPTERA, SILVANIDAE)* G. R. F. DAVIS Entomoiogy
Section, Research Station, Research Branch, Canada Agriculture, Saskatoon, Saskatchewan, Canada (Received 15 November
1960)
Abstract-Larvae of the saw-toothed grain beetle, Oryxaephilus surinamtxsis (L.), survived in greater numbers and developed quicker as the concentration of L-lysine in the diet increased from 0 to I.37 per cent. At higher concentrations the number surviving and the rate of growth decreased. Of related compounds tested, pipecolic acid, cr-aminoadipic acid, or a mixture of or-ketoglutaric acid and ammonium acetate partiaily supported growth; glutaric acid, ar-ketoglutaric acid, ammonium acetate, dipheny~hydantoin, or hydantoin did not.
INTRODUCTION LYSINE
is considered an essential amino acid for most organisms. However, it can be formed from aspartic acid by Esche-richia cdi (BOLTON et aZ., 1953) ; is essential for Streptococcus faecalis (6057) only under certain conditions (MCCLURE et al., 1954); and is synthesized by ~~tte~~a g~rn~~i~~ L., but not rapidly enough to satisfy the requirements of the organism (GORDON, 1959). Extensive reviews on lysine requirements and metabolism have been published by WORK (19%) and MEISTER (1957). DAVIS (1956) indicated that Oryzaephilus surinamensis (L.) could possibly dispense with lysine in the diet, At the time, because of the nature of the diet, absolute requirements of this organism for amino acids could not be determined. A chemically defined diet in which lysine can be a limiting amino acid has since been formulated. This is a report on the quantitative dietary requirements of larvae of 0. surinam~~~ for lysine. The effects of related compounds as replacements for lysine were also studied. MATERIALS
AND METHODS
The
components of the chemically defined diet were the same as those used previously (DAVIS, 1959) except that L-alanine and L-proline were included at the O-07 per cent level. VVith the same method as in earlier work (DAVIS, 1959), the effects of L-lysine were tested at twelve concentrations from 0 to 2.39 per cent. All constituents of these diets were obtained from Nutritional Biochemicals Corporation, Cleveland, Ohio. * Contribution
No. 65, Canada Department
of Agriculture
122
Research Station, Saskatoon, Sask.
LYSINE REQUIREMENTS OF LAEVAEOF ORYZAEPHILUS
SURINAMENSL!3
(L.)
123
The following related compounds were substituted for lysine in the diet: Dr..-pipecolic acid hydrochloride* at two concentrations; DL-~-a~noadipic acid* at four; and glutaric acid,$ a-ketoglutaric acidx (alone and with ammonium acetates), ammonium acetate, diphenylhydantoin,” and hydantoinf at one concentration each. RESULTS
AND DISCUSSION
Table 1 shows that as the percentage of lysine in the diet of the saw-toothed grain beetle increases, both survival and rate of development increase until a m~mum is reached at a.concentration of 1.37 per cent. As the concentra~on of iysine is further increased, survival and rate of development decrease rapidly. TABLE ~-THE NUMBER OF INDIVIDUALSPUPATING AND EMERGING AS ADULTS, AND THE AVERAGETIMESTo PUPATIONAND EMERGENCE WHEN 100 LARVAEOF Oryzaephilus sutinamensis WERE REAREDINDIVIDUALLYON RIETS CONTAININGVARIOUSQUANTITIESOF L-LYSINE Percentage L-lysine in diet ~-
Number pupating
::: 0.02 0.04 0.08 0.17 0.34 068 1.37 1‘71 2*05 2.39 LSD. at 5% level
; It 27 :: 70 74 27 :: 29
Average time (hr)
1764 1393 * 1353k 1227f 13.59 + 943 * 822 + 743+ 707* 1048+ 1002+ 1291 + -
144* 97 77 100 33 29 17 Ifs? 21 48 76
Number emerging ii ii: 20 :: t: 19 f 22
Average time (hr) 1884 1532 + 141* 1382+115 1336+ 84 1501+112 1066+ 38 968k 33 8632 19 788+ 1st 1169f 43 1122rt 54 1.5112 97 -
* Standard error. f Significantly less than other values at the 5 per cent level of confidence.
gos~~~ilu (Saund.) (V~DE~ANT, 1958); Apis rne~~~c~ L. (DE 1953) ; Aedes ffeg~p~~(L.) (GOLBERG and DE MEILLON, 1948 ; SINGH and BROWN, 1957) ; Calliphoru erythrocephalu (Meig.) (&DEE, 1954) ; Phormiu regina (Meig.) (MCGINNIS et al., 1956; CHELDELIN and NEWBURGH, 1959); Pseudosarcophaga a$%& (Fall.) (HOUSE, 1954); ‘Attugenus sp.’ (MOORE, 1946); Tribolium confusum DIN. (LEMONDE and BERNARD, 1951; FRAENKEL and PRINTY, 1954) ; and Trogoderma gvanarium Everts (PANT et al., 1958) require lysine for growth and development. The present work shows a definite requirement for lysine in the diet of the saw-toothed grain beetle. The results of earlier work (DAVIS, 1956) which indicated that 0. ~7~na~~ could dispense with lysine in the diet may Pec~~n~~~u
GROOT,
* Mann Research Laboratories, New York, N.Y. t Eastman Organic Chemicals, Rochester, N.Y. $ Nutritional Biochemicals Corporation, Cleveland, Ohio. 5 Fisher Scientific Company, Fairlawn, N. J.
G. R. F. DAVIS
124
have occurred because of impurities in the amino acid constituents (GORDON, 1959) or because of the presence of micro-organisms in the diet capable of converting aspartic acid to lysine (BOLTON et al., 1953). Table 2 indicates that larvae of 0. surinamer~+ on a lysine deficient diet may be able to utilize pipecolic acid, or-aminoadipic acid, or a mixture of or-ketoglutaric acid and ammonium acetate. These compounds either increased the rate of development or the survival of larvae reared without lysine. Glutaric acid, ol-ketoglutaric acid, ammonium acetate, diphenylhydantoin, or hydantoin did not. TABLE ~-THE NUMBEROF INDIVIDUALS PUPATINGAND EMERGINGAS ADULTS,AND THE AVERAGE TIMESTO PUPATIONANDEMERGENCE WHEN100 LARVAE OF Oryzaephilus surinamensis WRRE
REARED
INDIVIDUALLY
ON
DIRTS
COMPOUNDS
Lysine replaced by (%)
.
Number pupating
IN
-
_-
WHICH
VARIOUS
REPLACED
L-LYSINE
CONCENTRATIONS
OF
RELATED
-
Average time (hr)
Number
--
,emerging
Average time (hr)
DL-Pipecolic acid HCI
I.24 2.48
nr_-ar-Aminoadipic acid 1.21 2.42 3.63 4.84 Glutaric acid
l-37 c+Ketoglutaric acid 1.37 cY-Ketoglutaric acid + ammonium acetate 0.46 + 0.92 Ammonium acetate 1.37 Diphenylhydantoin 2.06 Hydantoin 0.94
2 0
972 + 34* 1156 &46 1150+53 -
1; :
2
1128 f26* -
7 10 -
1353 +45 1272 k47 -
-
0
-
0
-
18
11
1583 +93
1
1332
0
-
1740f125
1
0
-
1428
-
-
-
-
-
* Standard error.
The proposed pathway of lysine metabolism in the rat is as follows: lysine -+cu-aminoadipate+glutarate+ c+ketoglutarate and acetate (ROTHSTEIN and between lysine and or-aminoMILLER, 1954). MEISTER (1957) inserts pipecolate adipate. The results of the foregoing experiments suggest that this pathway is only partly operative in larvae of 0. Suriname&s, because the effective compounds substituted for lysine did not permit either as rapid development or as great survival as lysine itself. For the same reasons, the results indicate that these compounds, in the concentrations used, could not substitute entirely for lysine in the diet of 0. surinamensis. Acknowledgement-The acknowledged.
technical
assistance
of
Mr.
F.
T.
EVES is
gratefully
LYSINEREQUIRsMENTs OF LAnVAEOF ORYZAEPHILUS SLJRIiVAMENS1.S (L.)
12.5
REFE~NC~S BOLTONE. T., BRITTENR., and COWIE D. B. (1953) Synthesis of the aspartic and glutamic families of ammo acids by Escherichia coli. Science 17, 465. CHBLDELINV. H. and NEWBURGH R. W. (1959) Nutritional studies on the blowfly. In: Axenic culture of intervertebrate metazoa: a goal. Ann. N. Y. Acad. Sei. 77, 373-383. DAVIS G. R. F. (1956) Amino acid requirements of Ory,zuephiZus surirramensis (L.) (Coleoptera : Silvanidae) for pupation. Canad. J. 2001. 34, 82-85. DAVISG. R. F. (1959) Alanine and proline in the diet of larvae of Oryzaephilus su~~rn~~ (L.) (Coleoptera : Silvanidae). Arm. cnt. Sot. Ames. 52, 164-167. DE GROOTA. P. (1953) Protein and amino acid requirements of the honeybee (A&s mellijka L.). Physiol. camp. 3, l-89. FRAENKELG. and PRINTY G. E. (1954) The amino acid requirements of the confused flour beetle, Tribolium confusum Duval. B&Z. Bull., Woo& Hole 106, 149-157, GOLBERG L. and DE MEILLON B. (1948) The nutrition of the larvae of Aedes aegypti Linnaeus-IV. Protein and amino acid requirements. Bio&em. J. 43, 379-387. GORDONH. T. (1959) Minimal nutritional requirements of the German roach, Blattella germanica L. In: Axenic culture of invertebrate metazoa: a goal. Ann. N. Y. Acad. Sci. 77, 290-351. HOUSE H. L. (1954) Nutritional studies with Pseudasurcophaga a#+& (Fall.), a dipterous parasite of the spruce budworm, Choristonewu fzm$erana (Clem.)-HI. Effects of nineteen amino acids on growth. Canad, J. Zool. 32, 351-357. LEMQNDEA. and BERN- R. (1951) Nutrition des larves de Tribolium confusum DuvalII. Importance des acides amines. Car&-J. 2001. 29,80-83. MCCLURE L. E., NEUMANR. E., and MCCOY T. A. (1954) Amino acid metabolic studiesVI. Aspartic acid-lysine interrelations in Streptococcus faecalis (6057). Arch. Biochem. Bio@hysr. 53, 50-55. MCGINNIS A. J., NEW~URGHR. W., and CHELDELINV. H. (1956) Nutritional studies on the blowfly, F%orm& reg& (Meig.). g. N&r. S8, 309-324. MKIS-IERA. (1957) Bhhhtry of the Amino Acids. Academic Press, New York. MOOREW. (1946) Nutrition of Attagenus (I) sp. Ann. ent. Sot. Amer. 39, 513-521. PANT N. C., NAYARJ. K., and GUPTA P. (1958) On the significance of amino acids in the larval development of Khapra-beetle, Trogoderma granarium Everts. (Coleoptera : Dermestidae). E~&etttia 14, 176-178. ROTHSTEINM. and MILLER L. L. (1954) The metabolism of r&sine-6-C14. J. biol. Cha. 206, 243-253. SEDEE J. W. (1954) Qualitative amino acid requirements of larvae of Calliphora erythro.. cephala (Meigen). Acta n&l. phawn. physibl. 3, 262-269. SINGH K. R. P. and BROU~NW. A. (1957) Nutritional requirements of Aedes aegypti L. J. Ins. Physiol. 1, 199-220. VANDEIUANT E. A. (1958) The amino acid requirements of the pink bollworm. g. econ. Ent. 51, 309-311. WORK E. (1955) Some comparative aspects of lysine metabolism. A Sympoti~m on Amino Acid Met&o&m (Ed. by W. E. MCELROY and B. GLASS), pp. 462492. The Johns Hopkins Press, Baltimore.