U.S. patent application number 13/373562 was filed with the patent office on 2012-06-14 for food composition for hemophagous insects.
This patent application is currently assigned to TOKITAE LLC, a limited liability company of the State of Delaware. Invention is credited to E. Barcin Acar, Geoffrey F. Deane, 3ric Johanson, Emma Rae Mullen, Nathan P. Myhrvold, Nels R. Peterson, Clarence T. Tegreene, Charles Whitmer, Lowell L. Wood, JR..
Application Number | 20120148705 13/373562 |
Document ID | / |
Family ID | 45787879 |
Filed Date | 2012-06-14 |
United States Patent
Application |
20120148705 |
Kind Code |
A1 |
Acar; E. Barcin ; et
al. |
June 14, 2012 |
Food composition for hemophagous insects
Abstract
A composition suitable for feeding hemophagous insects includes
peptides, salt, and a CO.sub.2 generator.
Inventors: |
Acar; E. Barcin; (Sammamish,
WA) ; Deane; Geoffrey F.; (Bellevue, WA) ;
Johanson; 3ric; (Seattle, WA) ; Mullen; Emma Rae;
(Seattle, WA) ; Myhrvold; Nathan P.; (Bellevue,
WA) ; Peterson; Nels R.; (Seattle, WA) ;
Tegreene; Clarence T.; (Bellevue, WA) ; Whitmer;
Charles; (North Bend, WA) ; Wood, JR.; Lowell L.;
(Bellevue, WA) |
Assignee: |
TOKITAE LLC, a limited liability
company of the State of Delaware
Bellevue
WA
|
Family ID: |
45787879 |
Appl. No.: |
13/373562 |
Filed: |
November 17, 2011 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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12928460 |
Dec 10, 2010 |
8133524 |
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13373562 |
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Current U.S.
Class: |
426/1 ; 426/561;
426/562 |
Current CPC
Class: |
A23K 10/24 20160501;
A23K 20/22 20160501; A23K 50/90 20160501; A23K 20/147 20160501 |
Class at
Publication: |
426/1 ; 426/561;
426/562 |
International
Class: |
A23K 1/04 20060101
A23K001/04; A23K 1/16 20060101 A23K001/16; A23K 1/18 20060101
A23K001/18 |
Claims
1.-26. (canceled)
27. A food composition, comprising: a peptide component including
amino acids in the following dry-weight proportions: about
6.4%-about 6. 5% isoleucine; about 8.6%-about 9.0% leucine; about
6.2%-about 6.8% lysine; about 5.8%-about 6.4% phenylalanine; about
3.9%-about 4.5% threonine; about 1.1%-about 1.3% tryptophan; and
about 6.9%-about 7.2% valine; sodium chloride; and a CO.sub.2
generator, wherein the food composition is characterized in that
upon dilution to an isotonic solution, the peptide component is
present in a concentration of about 40 g/L to about 100 g/L.
28. The food composition of claim 27, wherein the peptide component
further comprises: about 4.5%-about 5.0% arginine.
29. The food composition of claim 27, wherein the peptide component
further comprises: about 1.5%-about 1.7% cysteine.
30. The food composition of claim 27, wherein the peptide component
further comprises: about 2.2% histidine.
31. The food composition of claim 27, wherein the peptide component
further comprises: about 4.3%-about 4.8% methionine.
32. The food composition of claim 27, wherein the peptide component
consists essentially of individual amino acids.
33. The food composition of claim 27, wherein the peptide component
includes oligopeptides.
34. The food composition of claim 27, wherein the peptide component
includes polypeptides.
35. (canceled)
36. (canceled)
37. The food composition of claim 27, wherein the CO.sub.2
generator includes an edible acid.
38. (canceled)
39. The food composition of claim 27, wherein the CO.sub.2
generator includes sodium bicarbonate.
40. The food composition of claim 27, further comprising a
sugar.
41. (canceled)
42. The food composition of claim 27, further comprising a blood
component.
43. (canceled)
44. The food composition of claim 42, wherein the blood component
is present at a concentration approximately equal to a normal
physiological level for human blood.
45. The food composition of claim 27, further comprising a
preservative.
46. The food composition of claim 27, further comprising a
stabilizer.
47. The food composition of claim 27, further comprising an insect
attractant, an insect repellent, a pheromone, a kairomone, an
allomone, or an insect phagostimulant.
48. The food composition of claim 27, further comprising a
colorant.
49. (canceled)
50. A food composition, comprising the food composition of claim 27
dissolved in a liquid.
51.-56. (canceled)
57. The food composition of claim 50, wherein the concentration of
the protein mixture in the liquid is from about 40 g/L to about 100
g/L.
58. The food composition of claim 50, wherein the liquid is
equivalently isotonic.
59. The food composition of claim 50, wherein the liquid is
substantially isotonic.
60. The food composition of claim 50, wherein the liquid is
isotonic.
61. (canceled)
62. A food composition, comprising: a peptide component including
amino acids isoleucine, leucine, lysine, phenylalanine, threonine,
tryptophan, and valine, these amino acids having relative
proportions substantially equal to the proportions that the same
amino acids would have in a mixture of albumin and whey protein at
a ratio between 3:1 and 12:1 dry weight; sodium chloride; and a
CO.sub.2 generator, the food composition characterized in that,
upon mixing with water to form an isotonic solution, the resulting
solution includes about 40 to about 100 grams of peptide component
per ml.
63. The food composition of claim 62, wherein the peptide component
is composed principally of amino acids.
64. The food composition of claim 62, wherein the peptide component
includes oligopeptides.
65. The food composition of claim 62, wherein the peptide component
includes polypeptides.
66.-67. (canceled)
68. The food composition of claim 62, wherein the CO.sub.2
generator includes an edible acid.
69. (canceled)
70. The food composition of claim 62, wherein the CO.sub.2
generator includes sodium bicarbonate.
71. The food composition of claim 62, further comprising a
sugar.
72. (canceled)
73. The food composition of claim 62, further comprising a blood
component.
74. (canceled)
75. The food composition of claim 73, wherein the blood component
is present at a concentration approximately equal to a normal
physiological level for human blood.
76. The food composition of claim 62, further comprising a
preservative.
77. The food composition of claim 62, further comprising a
stabilizer.
78. The food composition of claim 62, further comprising an insect
attractant, an insect repellent, a pheromone, a kairomone, an
allomone, or an insect phagostimulant.
79. The food composition of claim 62, further comprising a
colorant.
80. The food composition of claim 62, wherein the food composition
is water-soluble.
81. A food composition, comprising the food composition of claim 62
dissolved in a liquid.
82.-87. (canceled)
88. The food composition of claim 81, wherein the concentration of
the protein mixture in the liquid is from about 40 g/L to about 100
g/L.
89.-92. (canceled)
93. A food composition, comprising: a first portion including an
edible acid; and a second portion including sodium bicarbonate, the
first portion being separate from the second portion, wherein at
least one of the first portion and the second portion includes a
peptide component including amino acids isoleucine, leucine,
lysine, phenylalanine, threonine, tryptophan, and valine, these
amino acids having relative proportions substantially equal to the
proportions that the same amino acids would have in a mixture of
albumin and whey protein at a ratio between about 3:1 and about
12:1 dry weight, the food composition characterized in that, upon
mixing of the first portion, second portion, and optionally water
to form a mixture equivalently isotonic with blood, the edible acid
and the sodium bicarbonate combine to produce CO.sub.2.
94. The food composition of claim 93, where the peptide component
is present in an amount sufficient that upon mixing the first
portion, the second portion, and optionally water to form a mixture
equivalently isotonic with blood, the peptide component is present
in a concentration of about 40-100 g/L.
95. The food composition of claim 93, where the peptide component
is present in an amount sufficient that upon mixing the first
portion and the second portion of the food composition with an
amount of water sufficient to render the mixture substantially
isotonic with blood, the peptide component is present in a
concentration of about 40-100 g/L.
96. The food composition of claim 93, where the peptide component
is present in an amount sufficient that upon mixing the first
portion and the second portion of the food composition with an
amount of water sufficient to render the mixture isotonic with
blood, the peptide component is present in a concentration of about
40-100 g/L.
97. (canceled)
98. The food composition of claim 93, further comprising a
sugar.
99. (canceled)
100. The food composition of claim 93, further comprising a blood
component.
101. (canceled)
102. The food composition of claim 100, wherein the blood component
is present at a concentration approximately equal to a normal
physiological level for human blood.
103.-165. (canceled)
Description
SUMMARY
[0001] In one aspect, a food composition includes a protein mixture
including albumin and whey protein in a ratio between about 3:1 and
about 12:1 dry weight, respectively, sodium chloride, and a
CO.sub.2 generator. The CO.sub.2 generator may include sodium
bicarbonate or an edible acid (e.g., citric acid, acetic acid,
ascorbic acid, malic acid, lactic acid, tartaric acid, phosphoric
acid, oxalic acid, benzoic acid, or butyric acid). The food
composition may further include a sugar (e.g., glucose), a blood
component (e.g., plasma, hemoglobin, gamma globulin, red blood
cells, adenosine triphosphate, glucose, or cholesterol), which may
be at a concentration approximately equal to a physiological level
for human blood, a preservative, a stabilizer, an insect
attractant, an insect repellent, a pheromone, a kairomone, an
allomone, an insect phagostimulant, or a colorant. The food
composition may be water-soluble, and may be dissolved in a liquid
(e.g., water or blood plasma) or a gel, which may include a
preservative, a stabilizer, an insect attractant, an insect
repellent, a pheromone, a kairomone, an allomone, an insect
phagostimulant. Protein may be present in the liquid in a
concentration between about 40 g/L and 100 g/L, and the liquid may
be equivalently isotonic, substantially isotonic, or isotonic.
[0002] In another aspect, a food composition includes a peptide
component, sodium chloride, and a CO.sub.2 generator. The peptide
component includes amino acids in dry-weight proportions of about
6.4%-about 6. 5% isoleucine, about 8.6%-about 9.0% leucine, about
6.2%-about 6.8% lysine, about 5.8%-about 6.4% phenylalanine, about
3.9%-about 4.5% threonine, about 1.1%-about 1.3% tryptophan, and
about 6.9%-about 7.2% valine. The food composition is characterized
in that upon dilution to an isotonic solution, the peptide
component is present in a concentration of about 40 g/L to about
100 g/L. The peptide component may further include about 4.5%-about
5.0% arginine, about 1.5%-about 1.7% cysteine, about 2.2%
histidine, or about 4.3%-about 4.8% methionine. The peptime
component may consist essentially of individual amino acids, or may
include oligopeptides or polypeptides. The peptide component may
include albumin or whey protein. The CO.sub.2 generator may include
sodium bicarbonate or an edible acid (e.g., citric acid, acetic
acid, ascorbic acid, malic acid, lactic acid, tartaric acid,
phosphoric acid, oxalic acid, benzoic acid, or butyric acid). The
food composition may further include a sugar (e.g., glucose), a
blood component (e.g., plasma, hemoglobin, gamma globulin, red
blood cells, adenosine triphosphate, glucose, or cholesterol),
which may be at a concentration approximately equal to a
physiological level for human blood, a preservative, a stabilizer,
an insect attractant, an insect repellent, a pheromone, a
kairomone, an allomone, an insect phagostimulant, or a colorant.
The food composition may be water-soluble, and may be dissolved in
a liquid (e.g., water or blood plasma) or a gel, which may include
a preservative, a stabilizer, an insect attractant, an insect
repellent, a pheromone, a kairomone, an allomone, an insect
phagostimulant. Protein may be present in the liquid in a
concentration between about 40 g/L and 100 g/L, and the liquid may
be equivalently isotonic, substantially isotonic, or isotonic.
[0003] In another aspect, a food composition includes a peptide
component, sodium chloride, and a CO.sub.2 generator. The peptide
component includes amino acids isoleucine, leucine, lysine,
phenylalanine, threonine, tryptophan, and valine in a relative
proportion substantially equal to the proportions the same amino
acids would have in a mixture of albumin and whey protein at a
ratio between about 3:1 and about 12:1 dry weight. The food
composition is characterized in that, upon mixing with water to
form an isotonic solution, the resulting solution includes about 40
to about 100 grams of peptide component per ml. The peptide
component may be composed principally of amino acids, or may
include oligopeptides or polypeptides. The peptide component may
include albumin or whey protein. The CO.sub.2 generator may include
sodium bicarbonate or an edible acid (e.g., citric acid, acetic
acid, ascorbic acid, malic acid, lactic acid, tartaric acid,
phosphoric acid, oxalic acid, benzoic acid, or butyric acid). The
food composition may further include a sugar (e.g., glucose), a
blood component (e.g., plasma, hemoglobin, gamma globulin, red
blood cells, adenosine triphosphate, glucose, or cholesterol),
which may be at a concentration approximately equal to a
physiological level for human blood, a preservative, a stabilizer,
an insect attractant, an insect repellent, a pheromone, a
kairomone, an allomone, an insect phagostimulant, or a colorant.
The food composition may be water-soluble, and may be dissolved in
a liquid (e.g., water or blood plasma) or a gel, which may include
a preservative, a stabilizer, an insect attractant, an insect
repellent, a pheromone, a kairomone, an allomone, an insect
phagostimulant. Protein may be present in the liquid in a
concentration between about 40 g/L and 100 g/L, and the liquid may
be equivalently isotonic, substantially isotonic, or isotonic.
[0004] In another aspect, a food composition includes a first
portion including an edible acid (e.g., citric acid, acetic acid,
ascorbic acid, malic acid, lactic acid, tartaric acid, phosphoric
acid, oxalic acid, benzoic acid, or butyric acid), a second portion
separate from the first portion containing sodium bicarbonate. At
least one of the first and second portions includes a peptide
component including amino acids isoleucine, leucine, lysine,
phenylalanine, threonine, tryptophan, and valine, these amino acids
having relative proportions substantially equal to the proportions
of a mixture of albumin and whey protein at a ratio between about
3:1 and about 12:1 dry weight. The peptide component may be present
in an amount sufficient that upon mixing the first portion, the
second portion, and optionally water to form a mixture equivalently
isotonic with blood (or substantially isotonic with blood, or
isotonic with blood), the peptide component is present in a
concentration of about 40-100 g/L. The food composition may further
include a sugar (e.g., glucose), a blood component (e.g., plasma,
hemoglobin, gamma globulin, red blood cells, adenosine
triphosphate, glucose, or cholesterol), which may be at a
concentration approximately equal to a physiological level for
human blood, a preservative, a stabilizer, an insect attractant, an
insect repellent, a pheromone, a kairomone, an allomone, an insect
phagostimulant, or a colorant.
[0005] In another aspect, a method of feeding an insect includes
providing a solution including a mixture of albumin and whey
protein in a ratio between about 3:1 and about 12:1, respectively,
sodium chloride, water, and a source of carbon dioxide, and placing
the solution in a container including a barrier surface, wherein
the barrier surface is configured to be penetrated by the insect to
feed upon the solution. The method may further include
substantially maintaining the solution at a selected temperature,
or inducing convection (e.g., free or forced convection) in the
solution; The method may further include exposing the solution to
an insect such as a mosquito (e.g., of genus Anopheles, Aedes, or
Culex). At least one of the barrier surface, the aqueous solution,
and the container may include an insect attractant, an insect
repellent, a pheromone, a kairomone, an allomone, or an insect
phagostimulant. The source of carbon dioxide may be a chemical
reaction (e.g., a reaction between sodium bicarbonate and an edible
acid such as citric acid, acetic acid, ascorbic acid, malic acid,
lactic acid, tartaric acid, phosphoric acid, oxalic acid, benzoic
acid, or butyric acid). The solution may further include a sugar
(e.g., glucose), a blood component (e.g., plasma, hemoglobin, gamma
globulin, red blood cells, adenosine triphosphate, glucose, or
cholesterol), which may be at a concentration approximately equal
to a physiological level for human blood, a preservative, a
stabilizer, or a colorant. The concentration of protein in the
solution may be from about 40 g/L to about 100 g/L. The solution
may be equivalently isotonic, substantially isotonic, or
isotonic.
[0006] In another aspect, a method of feeding an insect includes
providing a solution including a peptide component, sodium
chloride, water, and a source of carbon dioxide, and placing the
solution in a container including a barrier surface, wherein the
barrier surface is configured to be penetrated by the insect to
feed upon the solution. The peptide component includes amino acids
isoleucine, leucine, lysine, phenylalanine, threonine, tryptophan,
and valine, these amino acids having relative proportions
substantially equal to the proportions the same amino acids would
have in a mixture of albumin and whey protein at a ratio between
about 3:1 and about 12:1 dry weight. The peptide component may be
composed principally of amino acids, or may include oligopeptides
or polypeptides. The peptide component may include albumin or whey
protein. The method may further include substantially maintaining
the solution at a selected temperature, or inducing convection
(e.g., free or forced convection) in the solution. The method may
further include exposing the solution to an insect such as a
mosquito (e.g., of genus Anopheles, Aedes, or Culex). At least one
of the barrier surface, the aqueous solution, and the container may
include an insect attractant, an insect repellent, a pheromone, a
kairomone, an allomone, or an insect phagostimulant. The source of
carbon dioxide may be a chemical reaction (e.g., a reaction between
sodium bicarbonate and an edible acid such as citric acid, acetic
acid, ascorbic acid, malic acid, lactic acid, tartaric acid,
phosphoric acid, oxalic acid, benzoic acid, or butyric acid). The
solution may further include a sugar (e.g., glucose), a blood
component (e.g., plasma, hemoglobin, gamma globulin, red blood
cells, adenosine triphosphate, glucose, or cholesterol), which may
be at a concentration approximately equal to a physiological level
for human blood, a preservative, a stabilizer, or a colorant. The
concentration of the peptide component in the solution may be from
about 40 g/L to about 100 g/L. The solution may be equivalently
isotonic, substantially isotonic, or isotonic.
[0007] The foregoing summary is illustrative only and is not
intended to be in any way limiting. In addition to the illustrative
aspects, embodiments, and features described above, further
aspects, embodiments, and features will become apparent by
reference to the drawings and the following detailed
description.
DETAILED DESCRIPTION
[0008] As used herein unless context dictates otherwise, the terms
"feeding medium" and "food composition" include compositions
suitable for feeding hemophagous insects. As used herein unless
context dictates otherwise, the term "artificial diet" includes
food compositions other than whole blood of a person or animal.
Artificial diets may include blood components, such as but without
limitation plasma, hemoglobin, or red blood cells. Food
compositions, feeding media, or artificial diets may, but need not,
support egg-laying in hemophagous insects.
[0009] When raising hemophagous insects (e.g., for research
purposes), it may be inconvenient or expensive to provide blood
meals. In some cases, insects may be kept on artificial diets which
may be more readily stored and handled in a laboratory. In some
cases, an artificial diet (e.g., glucose water or water-soaked
raisins) has been found to be adequate for maintaining health of
hemophagous insects (e.g., mosquitoes), but it has not been
adequate in order to stimulate egg-laying.
[0010] While the following discussion is in the context of feeding
of mosquitoes, the formulations described herein, as well as the
methods of determining appropriate formulations, are expected to be
applicable to other hemophagous insects as well (e.g., tsetse
flies, lice, bed bugs, no-see-urns, fleas, sand flies, midges,
snipe flies, horse flies, stableflies, or sheep flies).
[0011] Uchida describes a set of seven "absolutely essential" amino
acids for stimulating oogenesis in Culex pipiens pallens mosquitoes
(isoleucine, leucine, lysine, phenylalanine, threonine, tryptophan,
and valine), and identifies four more that "may be essential" or
"are required for a full level of initiation and promotion of egg
development" (arginine, cysteine, histidine, and methionine). See,
Uchida, "Balanced. Amino Acid Composition Essential for
Infusion-induced Egg Development in the Mosquito (Culex pipiens
pallens)," J. Insect Physiol., 39(7):615-621 (1993). See also,
Uchida, et al., "Ovarian development induced in decapitated female
Culex pipiens pallens mosquitoes by infusion of physiological
quantities of 20-hydroxyecdysone together with amino acids," J.
Insect Physiol., 44:525-528 (1998) (stimulating oogenesis in
decapitated females with a combination of 20-hydroxyecdysone
hormone and a mixture of 17 amino acids). Chang, et al. found that
supplementation with additional isoleucine enhanced egg production
rate in Aedes aegypti mosquitoes fed with human red blood cells.
See, Chang et al., "Amino Acid Composition of Human and Guinea Pig
Blood Proteins, and Ovarian Proteins of the Yellow Fever Mosquito
Aedes aegypti; and Their Effects on the Mosquito Egg Production,"
Comp. Biochem. Physiol., 62A:753-755 (1979). See also, Greenberg,
"Some Nutritional Requirements of Adult Mosquitoes (Aedes aegypti)
for Oviposition," J. Nutr. 43(1):27-35 (1951) (isoleucine
supplementation enhanced egg-laying after blood feeding). Arsic, et
al., found that albumin-containing substances would support
follicular development in Anopheles gambiae mosquitoes, but did not
support complete egg development without addition of red blood
cells in this species. Griffith, et at, were able to support a
population of Culex quinquefasciatus mosquitoes for more than fifty
generations on a blood substitute artificial diet including
ovalbumin, soya infant formula, globulins, and adenosine
triphosphate. See, "Culturing Culex quinquefasciatus mosquitoes
with a blood substitute diet for the females," Med. and Vet.
Entomology 10:265-268 (1996). Each of the above-mentioned
publications is incorporated by reference herein to the extent not
inconsistent herewith.
[0012] A population of Anopheles mosquitoes may be maintained
through an indefinite number of generations on our artificial
feeding medium without any blood feeds. Each of the experiments
described below used a combination of ovalbumin (Sigma, Grade II,
A5253, CAS 9006-59-1, containing 75%.+-.7% protein plus
carbohydrate and phosphate portions) and whey protein
(SIMPLESSE.TM. from CP Kelco, a microparticulated whey protein
concentrate containing 53% bovine whey protein plus food-grade
xantham gum). In some embodiments, the dry weight ratios of albumin
to whey protein may be about 1:5, about 1:4, about 1:3, about 1:2,
about 1:1, about 2:1, about 3:1, about 4:1, about 5:1, about 6:1,
about 7:1, about 8:1, about 9:1, about 10:1, about 11:1 or about
12:1, respectively. In addition, dry weight ratios in various
ranges between any of these values is contemplated. For example,
and without limitation, ratios of about 2.25:1, about 3.25:1, about
4.25:1, about 5.5:1, about 6.5:1, about 7.5:1, about 8.5:1, about
9:5:1, about 10.5:1, and about 11.5:1. For example, and without
limitation, dry weight ratios of albumin to whey protein may be
about 3:1 to about 6:1, respectively; or about 3:1 to about 12:1,
respectively. In addition to protein, in some embodiments a food
composition may contain other macronutrients such as carbohydrates
or lipids, or micronutrients such as vitamins or minerals.
[0013] Food compositions that are substantially isotonic with blood
(reference osmolality of 275-300 mOsm/kg) can help avoid insect
death by osmotic imbalance after feeding. Isotonicity can be
achieved, for example, by either adding Ringer's salts or the
formula of Griffith, et al. (cited above). As used herein, an
"isotonic" solution is one having a tonicity within .+-.5% of the
tonicity of blood (e.g., 5% less than bottom of reference range, 4%
less, 3% less, 2% less, 1% less, within reference range, 1% more
than top of reference range, 2% more, 3% more, 4% more, or 5%
more), while a "substantially isotonic" solution is one having an
tonicity within .+-.10% of the tonicity of blood (e.g., 10% less
than bottom of reference range, 9% less, 8% less, 7% less, 6% less,
6% more than top of reference range, 7% more, 8% more, 9% more, or
10% more). An "equivalently isotonic" solution has an tonicity
within .+-.20% of the tonicity of blood (e.g., 20% less than bottom
of reference range, 19% less, 18% less, 17% less, 16% less, 15%
less, 14% less, 13% less, 12% less, 11% less, 11% more than top of
reference range, 12% more, 13% more, 14% more, 15% more, 16% more,
17% more, 18% more, 19% more, or 20% more).
[0014] In addition to the protein mixtures discussed above,
attractants can be added to the food composition to increase
mosquito feeding rates. In some embodiments, an attractant is
CO.sub.2. CO.sub.2 can be produced within the food composition
either by mixing proteins and salts with carbonated "sparkling"
water, or by using a chemical CO.sub.2 generator, for example,
citric acid (or another acid edible by hemophagous insects, such as
ascorbic acid, malic acid, lactic acid, tartaric acid, phosphoric
acid, oxalic acid, benzoic acid, or butyric acid) mixed with sodium
bicarbonate. Some embodiments may use other CO.sub.2 generators;
for example, other chemical reactions that produce CO.sub.2,
bubbling CO.sub.2 through or blowing it onto a feeding medium, or
biological CO.sub.2 generators (e.g., yeast). In some embodiments,
the feeding medium may be in two separate parts, which can be mixed
together (e.g., at feeding time) to begin the CO.sub.2-generating
reaction (e.g., a reaction between an edible acid and sodium
bicarbonate). In some embodiments, other attractants may be
included (e.g., lactic acid, pentyl vinyl carbinol, or isolaveric
acid), or other insect-behavior-modifying ingredients may be used
(e.g., repellents that discourage undesired insects, pheromones,
allomones, kairomones, or phagostimulants).
Insect-behavior-modifying ingredients may, for example, encourage
or discourage feeding of a particular species, or of insects with
certain conditions (e.g., gravid females), or might encourage or
discourage crop feeding or midgut engorgement.
[0015] In addition to, or instead of, attractants and other
insect-behavior-modifying components, a food composition may
include other active ingredients that affect mosquito metabolism or
physiology. In one embodiment, a food composition may be used to
control mosquito population by including a toxin, parasite,
microorganism, or biocontrol agent (e.g., Ascogregarine
trophozoites, Bacillus thuringiensis strains, Bacillus sphaericus,
Wolbachia, Microsporidia such as Nosema sp., Coelomomyces sp.
fungi, Lagenidium, Saccharaopolyspora spinosa, Metarrhizium
anisopliae, Edhazardia aedes, Beauveria bassiana spores, and
mermithid nematodes). See, e.g., Reyes-Villanueva, et al.,
"Susceptibility of Aedes aegypti and Aedes albopictus larvae to
Ascogregarina culicis and Ascogregarina taiwanensis" J. Inverteb.
Path. 84:47-53 (2003); Saridaki, et al., "Wolbachia: more than just
a bug in insects genitals," Curr. Opinion in Microbiol. 13:67-72
(2010); Axtell, et al., "Encapsulation of the Mosquito Fungal
Pathogen Lagendium giganteum (Oomyetes:Lagenidiales) in Calcium
Alginate," J. Am. Mosq. Control Assoc. 3(3):450-459 (1978);
Lichtwardt, et al., "A new Coelomomyces pathogenic to mosquitoes in
Costa Rica," Rev. Biol. Trop. 41(3):407-410 (1993); Schenker, et
al., "The effects of Nosema algerae on the development of
Plasmodium yoelii nigeriensis in Anopheles stephensi," Parasitol.
Res. 78:56-59 (1992); Bukhari, et al., "Factors affecting
fungus-induced larval mortality in Anopheles gambiae and Anopheles
stephensi," Malaria J. 9(22):1-15 (2010); Perez, et al., "Spinosad,
a Naturally Derived Insecticide, for Control of Aedes aegypti
(Diptera: Culicidae): Efficacy, Persistence, and Elicited
Oviposition Response," J. Med. Entomol. 44(4):631-638 (2007);
Cetin, et al., "Evaluation of the naturally-derived insecticide
spinosad against Culex pipiens L. (Diptera: Culicidae) larvae in
septic tank water in Antalya, Turkey," J. Vect. Ecol. 30(1):151-154
(2005); Becnel, et al., "Influence of temperature on developmental
parameters of the parasite/host system Edhazardia aedis
(Microsporida: Amblyosporidae) and Aedes aegypti (Diptera:
Culicidae)," J. Invert. Pathol. 60(3):299-303 (1992); Petersen,
"Role of mermithid nematodes in biological control of mosquitoes,"
Exp. Parasitol. 33(2):239-247 (1973), each of which is incorporated
by reference herein to the extent not inconsistent herewith.
[0016] In some embodiments, it may also be useful to deliver an
agent to the insect. For example, for the study of malaria,
Plasmodium parasites (e.g., P. falciparum gametocytes) can be added
to the food composition. In some embodiments, the agent added can
be a toxin, parasite, a beneficial agent, etc. In some embodiments,
if toxins are added, the toxins may be sequestered (e.g., separated
from the food composition or from immediate contact with the
insect). In some such embodiments, these sequestered toxins may
become unsequestered over a period of time, for example upon return
to a breeding site, potentially killing immature insects (for
example, to control a population). For example, toxins (or other
agents) may include a dissolvable coating, or a cleavable blocking
agent, so that they can be ingested and then excreted in a location
where the coating will dissolve and release the toxin or other
agent. In some embodiments, the medium may include enzymes,
antibiotics, or antiseptics.
[0017] The feeding medium may also contain "inactive" ingredients
that do not act directly upon the mosquito metabolism, (e.g., by
providing nourishment or affecting a biological function of the
mosquito). For example, the formulations described below made with
SIMPLESSE.TM. include xantham gum. In some embodiments, this
ingredient may modify the texture or rheological properties of the
feeding medium. In some embodiments, other rheological agents may
be added or substituted, while some embodiments may not include
rheological agents. In some embodiments, the medium may include
stabilizers, preservatives, colorants, pH modifiers, or other
components that do not act directly upon the mosquito metabolism.
In some embodiments, the medium may include blood components, such
as plasma, hemoglobin, gamma globulin, red blood cells, adenosine
triphosphate (ATP), glucose, or cholesterol. These ingredients may
be added for their physical effects on the medium, or for their
influence on insect behavior.
[0018] In some embodiments the feeding medium is a liquid having
active or inactive ingredients dissolved therein. In some
embodiments, one or more of the ingredients may be in suspension.
In some embodiments, the feeding medium may be a gel, or may
include a mixture of phases (e.g., a liquid-soaked sponge).
[0019] In some embodiments, rather than using whole albumin or whey
proteins, these proteins may be fully or partially hydrolyzed. In
some embodiments, peptide mixtures may further include proportions
of any of the four additional amino acids that Uchida suggests "may
be essential" or "are required for a full level of initiation and
promotion of egg development." These proportions have been
calculated using the protein sequence of albumin (see, Nisbet, et
al., "The Complete Amino-Acid Sequence of Hen Ovalbumin," Eur. J.
Biochem., 115:335-345 (1981), which is incorporated by reference
herein) and an amino acid breakdown for whey protein isolate
published by Davisco Foods (see, "Whey Protein Concentrate 80%,"
Version 07F-0604, included herewith and incorporated by reference
herein). (The amino acid breakdown of SIMPLESSE.TM. was not readily
available, but is expected to be substantially similar to the
Davisco product). Table 1 shows a breakdown by weight percent of
amino acids of each of these proteins, as well as a calculated
value for ratios of 4.25:1 and 8.5:1 of albumin to whey
protein.
TABLE-US-00001 TABLE 1 Whey Albumin protein 3:1 4.25:1 8:5:1 12:1
wt % wt % albumin:whey albumin:whey albumin:whey albumin:whey
Leucine 8.43% 10.8% 9.0% 8.9% 8.7% 8.6% Valine 7.29% 5.8% 6.9% 7.0%
7.1% 7.2% Isoleucine 6.59% 5.8% 6.4% 6.4% 6.5% 6.5% Phenylalanine
6.64% 3.3% 5.8% 6.0% 6.3% 6.4% Lysine 5.87% 9.6% 6.8% 6.6% 6.3%
6.2% Threonine 3.59% 7.2% 4.5% 4.3% 4.0% 3.9% Tryptophan 1.09% 1.8%
1.3% 1.2% 1.2% 1.1% Methionine 5.09% 1.9% 4.3% 4.5% 4.8% 4.8%
Arginine 5.25% 2.1% 4.5% 4.6% 4.9% 5.0% Histidine 2.18% 2.2% 2.2%
2.2% 2.2% 2.2% Cysteine 1.46% 2.3% 1.7% 1.6% 1.5% 1.5% Serine 8.02%
4.7% 7.2% 7.4% 7.7% 7.8% Alanine 6.26% 4.9% 5.9% 6.0% 6.1% 6.2%
Glutamic acid 9.76% 16.7% 11.5% 11.1% 10.5% 10.3% Glycine 2.87%
1.8% 2.6% 2.7% 2.8% 2.8% Asparagine 4.55% unk unk unk unk unk
Glutamine 4.43% unk unk unk unk unk Aspartic acid 3.74% 10.8% 5.5%
5.1% 4.5% 4.3% Proline 3.24% 5.8% 3.9% 3.7% 3.5% 3.4% Tyrosine
3.64% unk unk unk unk unk
EXAMPLES
[0020] Each of the experiments described below used a combination
of ovalbumin (Sigma, Grade II, A5253, CAS 9006-59-1, containing
75%.+-.7% protein plus carbohydrate and phosphate portions) and
whey protein (SIMPLESSE.TM. from CP Kelco, a microparticulated whey
protein concentrate containing 53% bovine whey protein plus
food-grade xantham gum). Product information sheets for these two
products ("Albumin from chicken egg white," Product Number A 5253,
Sigma Aldrich Corporation, (2002); "CP Kelco Microparticulated Whey
Protein Concentrate," CP Kelco (2010), available at
www.cpkelco.com/products-mwpc.html) are incorporated herein by
reference.
Example #1
[0021] 3 grams of ovalbumin powder and 1 g of microparticulated
whey protein concentrate were dissolved into 50 ml of carbonated
Ringer's solution (made by dissolving 0.9 g NaCl, 0.042 g KCl, and
0.025 g CaCl.sub.2 into 100 ml of carbonated H.sub.2O). Carbonated
H.sub.2O was unflavored TALKING RAIN.TM. canned soda. The resulting
solution had a calculated ratio of ovalbumin to whey protein of
4.25, and a calculated protein concentration of 55.6 g/L (using the
protein fractions for Sigma-Aldrich ovalbumin powder and
SIMPLESSE.TM. microparticulated whey protein cited above). The
solution was offered to 34 mosquitoes in a membrane feeder heated
by water circulation, at an initial temperature of 38.degree. C. We
observed one of the mosquitoes (3%) to feed (become engorged) upon
the solution, but no eggs were detected.
Example #2
[0022] Example 2 was carried out at 38.degree. C..+-.0.5.degree. C.
(unless otherwise noted), using the membrane feeder described in
copending and commonly owned U.S. Application No. To be Assigned
(filed on even date herewith entitled INSECT FEEDER, naming E.
Barcin Acar, David R. Burton, Ted B. Ellis, Emma Rae Mullen, David
R. Nash, and Michael Vinton as inventors, which is incorporated by
reference herein). We dissolved 3.6 g of ovalbumin powder and 0.6 g
of microparticulated whey protein concentrate into 50 ml of
carbonated Ringer's solution. The resulting solution had a
calculated ratio of ovalbumin to whey protein of 8.49, and a
calculated protein concentration of 60.4 g/L. On two separate
occasions, this diet was offered to 41 mosquitoes and to 25
mosquitoes, and 6 fed on the first occasion (15%) and 5 fed on the
second occasion (20%). These mosquitoes produced eggs.
[0023] 0.03 g of ATP (the amount used by Griffith, et al., cited
above) were added to the above diet, and the resulting solution
offered to 40 mosquitoes in a membrane feeder heated by water
circulation, at an initial temperature of 38.degree. C. Two
mosquitoes (5%) were observed to feed, and no eggs were
detected.
[0024] 0.045 g of glucose were added to the above diet (without
ATP), corresponding to a normal human blood sugar of 90 mg/dL, and
the resulting solution offered to 36 mosquitoes in a membrane
feeder heated by water circulation, at an initial temperature of
38.degree. C. Three mosquitoes (8%) were observed to feed. These
mosquitoes produced eggs.
Example #3
[0025] We dissolved 8 g of ovalbumin powder, 0.6 g of globulin
powder, and 0.32 g of hemoglobin into 40 ml of carbonated Ringer's
solution. This solution was offered to 38 mosquitoes in a membrane
feeder heated by water circulation, at an initial temperature of
38.degree. C. Three mosquitoes (8%) were observed to feed, but no
eggs were detected.
Example #4
[0026] We dissolved 3 g of ovalbumin powder and 1 g of
microparticulated whey protein concentrate into 50 ml of carbonated
Griffith's solution (made by dissolving 0.6 g NaCl and 0.06 g of
NaHCO.sub.3 into 100 ml of carbonated H.sub.2O). The resulting
solution had a calculated ratio of ovalbumin to whey protein of
4.24, and a calculated protein concentration of 55.6 g/L. When
offered to 31 mosquitoes in a membrane feeder heated by water
circulation, at an initial temperature of 38.degree. C., we
observed one of the mosquitoes (3%) to feed, and this mosquito did
lay eggs.
[0027] 0.045 g of glucose were added to the above diet, and the
resulting solution offered to 41 mosquitoes. Six mosquitoes (15%)
were observed to feed. These mosquitoes produced eggs.
[0028] 0.045 g of glucose and 0.01 g of powdered CENTRUM.TM.
multivitamin were added to the above diet, and the resulting
solution offered to 30 mosquitoes. Four mosquitoes (13%) were
observed to feed, and these mosquitoes produced eggs. The exact
amount of multivitamin that was dissolved in the solution is
uncertain; vitamin particles were observed to settle at the bottom
of the feeder. The addition of vitamins was observed to increase
feeding rates in at least some cases, however. Suitable vitamin
levels for individual vitamins can be determined by routine
additional experimentation according to the techniques described
herein. Liquid vitamin formulations may be used to improve
solubility, although solid vitamins may also be used if adequately
dissolved or suspended.
Example #5
[0029] We dissolved 3.6 g of ovalbumin powder, 0.6 g of
microparticulated whey protein concentrate, and 0.045 g of glucose
into 50 ml of carbonated Griffith's solution. The resulting
solution had a calculated ratio of ovalbumin to whey protein of
8.49, and a calculated protein concentration of 60.4 g/L. When
offered to 37 mosquitoes in a membrane feeder heated by water
circulation, at an initial temperature of 38.degree. C., we
observed 8 of the mosquitoes (22%) to feed. These mosquitoes
produced eggs.
Example #6
[0030] For this and Example 7, we used 1.8 g of ovalbumin powder,
0.3 g of microparticulated whey protein concentrate, 0.1 g of NaCl,
0.185 g of sodium bicarbonate (NaHCO.sub.3), and 0.065 g of citric
acid (C.sub.6H.sub.8O.sub.7) dissolved in 20 ml of distilled water.
The resulting solution had a calculated ratio of ovalbumin to whey
protein of 8.49, and a calculated protein concentration of 75.5
g/L. The sodium bicarbonate and the citric acid react to form
sodium citrate and carbon dioxide, which is evolved from the
solution to serve as a mosquito attractant. On two separate
occasions, this diet was offered to 29 mosquitoes and to 44
mosquitoes using the membrane feeder described in Example #2. 17
fed on the first occasion (59%) and 26 fed on the second occasion
(59%). These mosquitoes produced eggs.
[0031] When 0.18 g of L-cysteine was added to the above solution
(giving a calculated protein concentration of 84.5 g/L) on three
occasions feeding rates were reduced: 17 of 39 mosquitoes (44%) fed
the first time, 4 of 32 mosquitoes (13%) fed the second time, and
15 of 29 mosquitoes (52%) fed the third time. The first group of
mosquitoes was not observed to produce eggs, but the second and
third groups did.
[0032] When 0.0225 g of glucose (corresponding to a normal blood
sugar of 112.5 mg/dL) were added to the above solution (without
L-cysteine), feeding rates were also reduced in five repetitions:
11 of 37 mosquitoes (30%), 14 of 32 mosquitoes (44%), 10 of 28
mosquitoes (36%), 20 of 37 mosquitoes (54%), and 18 of 36
mosquitoes (50%). All groups of mosquitoes produced eggs.
[0033] When 3.02 mg of lactic acid were added to the above solution
(without L-cysteine or glucose), 9 out of 20 mosquitoes (45%) were
observed to feed. These mosquitoes produced eggs.
[0034] When 1 ml of deionized water in the above solution (without
L-cysteine, glucose, or lactic acid) was replaced with 1 ml of
acetic acid, the feeding rate dropped to 6 out of 23 mosquitoes
(26%), but the mosquitoes still produced eggs. When 1 ml of
isovaleric acid was substituted instead of the acetic acid, 19 of
34 mosquitoes (56%) fed on a first occasion, and 9 of 35 mosquitoes
(26%) fed on a second occasion. These mosquitoes produced eggs.
Example #7
[0035] Groups of 200 mosquito pupae were randomly collected from
larval trays and placed in polycarbonate and polyethylene cages,
with access to water and raisins as a sugar source. Seven days
later, these mosquitoes were fed a solution consisting of 1.8 g of
ovalbumin powder, 0.3 g of microparticulated whey protein
concentrate, 0.1 g NaCl, 0.0225 g glucose, 0.065 g citric acid, and
0.185 g sodium bicarbonate in 20 ml of deionized water. These
mosquitoes laid eggs, which were hatched, and the larvae reared
according to the same regimen. These mosquitoes have been bred for
fifteen generations according to this protocol. Feeding rates are
about 52% for each generation, and the females produced about
31.5.+-.5.1 eggs/female over the course of the generations, with
the number increasing slightly in successive generations. While
these feeding and fecundity rates are lower than those for
blood-fed mosquitoes (64% and 58.+-.4.4 eggs/female in our
insectary), they are still adequate to maintain a continuing
mosquito population. Offspring of artificial medium-fed mosquitoes
were observed to be similar to those of blood-fed mosquitoes in
size, pupal weight, longevity, and sex ratio.
[0036] Various embodiments of insect feeding media and methods have
been described herein. In general, features that have been
described in connection with one particular embodiment may be used
in other embodiments, unless context dictates otherwise. For
example, the multivitamin found in Example #4 may optionally be
used in any other formulation. For the sake of brevity,
descriptions of such features have not been repeated, but will be
understood to be included in the different aspects and embodiments
described herein.
[0037] It will be understood that, in general, terms used herein,
and especially in the appended claims, are generally intended as
"open" terms (e.g., the term "including" should be interpreted as
"including but not limited to," the term "having" should be
interpreted as "having at least," the term "includes" should be
interpreted as "includes but is not limited to," etc.). It will be
further understood that if a specific number of an introduced claim
recitation is intended, such an intent will be explicitly recited
in the claim, and in the absence of such recitation no such intent
is present. For example, as an aid to understanding, the following
appended claims may contain usage of introductory phrases such as
"at least one" or "one or more" to introduce claim recitations.
However, the use of such phrases should not be construed to imply
that the introduction of a claim recitation by the indefinite
articles "a" or "an" limits any particular claim containing such
introduced claim recitation to inventions containing only one such
recitation, even when the same claim includes the introductory
phrases "one or more" or "at least one" and indefinite articles
such as "a" or "an" (e.g., "an insect" should typically be
interpreted to mean "at least one insect"); the same holds true for
the use of definite articles used to introduce claim recitations.
In addition, even if a specific number of an introduced claim
recitation is explicitly recited, it will be recognized that such
recitation should typically be interpreted to mean at least the
recited number (e.g., the bare recitation of "two insects," or "a
plurality of insects," without other modifiers, typically means at
least two insects). Furthermore, in those instances where a phrase
such as "at least one of A, B, and C," "at least one of A, B, or
C," or "an [item] selected from the group consisting of A, B, and
C," is used, in general such a construction is intended to be
disjunctive (e.g., any of these phrases would include but not be
limited to systems that have A alone, B alone, C alone, A and B
together, A and C together, B and C together, or A, B, and C
together, and may further include more than one of A, B, or C, such
as A.sub.1, A.sub.2, and C together, A, B.sub.1, B.sub.2, C.sub.1,
and C.sub.2 together, or B.sub.1 and B.sub.2 together). It will be
further understood that virtually any disjunctive word or phrase
presenting two or more alternative terms, whether in the
description, claims, or drawings, should be understood to
contemplate the possibilities of including one of the terms, either
of the terms, or both terms. For example, the phrase "A or B" will
be understood to include the possibilities of "A" or "B" or "A and
B." Moreover, "may," "can," "optionally," and other permissive
terms are used herein for describing optional features of various
embodiments. These terms likewise describe selectable or
configurable features generally, unless the context dictates
otherwise.
[0038] While various aspects and embodiments have been disclosed
herein, other aspects and embodiments will be apparent to those
skilled in the art. The various aspects and embodiments disclosed
herein are for purposes of illustration and are not intended to be
limiting, with the true scope and spirit being indicated by the
following claims.
* * * * *
References