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Luca Valzania, Vincent G. Martinson, Ruby E. Harrison, Bret M. Boyd, Kerri L. Coon, Mark R. Brown, Michael R. Strand
Both living bacteria and eukaryotes in the mosquito gut promote growth of larvae
www.plosntds.org
www.plosntds.org
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[()30()]TJ
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[()30()6()11()10(\r)6()15()4()8()]TJ
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[()2()16()17()11(\n)4()6()]TJ
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0.83 0.64 0.02 0 k
(30)Tj
0 g
(].)Tj
2.0806 0 Td
(Growth)Tj
3.3221 0 Td
(of)Tj
/F9 1 Tf
1.0148 0 Td
[()]TJ
/F8 1 Tf
(.)Tj
/F9 1 Tf
0.9921 0 Td
[()7()10()12(\n)]TJ
/F8 1 Tf
1.5647 0 Td
(or)Tj
/F9 1 Tf
1.0942 0 Td
[()]TJ
/F8 1 Tf
(.)Tj
/F9 1 Tf
0.9014 0 Td
[()7()2()20()-5()-13(\n)-4()10(\n)4()6()]TJ
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3.9118 0 Td
(was)Tj
1.7008 0 Td
(monitored)Tj
4.49 0 Td
(by)Tj
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(measuring)Tj
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(optical)Tj
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(density)Tj
-34.1118 -1.2983 Td
(\(OD\))Tj
2.3811 0 Td
(at)Tj
0.9525 0 Td
(600)Tj
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(nM)Tj
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(followed)Tj
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(by)Tj
1.1849 0 Td
(determination)Tj
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(of)Tj
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(cell)Tj
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(density)Tj
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(using)Tj
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(a)Tj
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(Neubauer)Tj
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(hemocytometer)Tj
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1.0262 0 Td
(generate)Tj
3.6056 0 Td
(growth)Tj
3.0727 0 Td
(curves)Tj
2.7836 0 Td
(for)Tj
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(each.)Tj
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(An)Tj
1.4514 0 Td
(OD)Tj
1.7064 0 Td
(of)Tj
1.0148 0 Td
(1.0)Tj
1.3946 0 Td
(equaled)Tj
3.3109 0 Td
(~1)Tj
1.2756 0 Td
(x)Tj
0.6859 0 Td
(10)Tj
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q
1 j
1 J
0 w
7.4999 0 0 6.6659 459.1559 484.1574 cm
BT
/F8 1 Tf
1 TL
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(9)Tj
ET
Q
q
1 j
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0.9978 0 Td
[()7()10()12(\n)]TJ
/F8 1 Tf
1.5591 0 Td
(and)Tj
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(~6)Tj
1.2756 0 Td
(x)Tj
0.686 0 Td
(10)Tj
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Q
q
1 j
1 J
0 w
7.4999 0 0 6.6659 229.096 471.1181 cm
BT
/F8 1 Tf
1 TL
-0.0053 Tc
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(7)Tj
ET
Q
q
1 j
1 J
0 w
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BT
/F8 1 Tf
1 TL
-0.004 Tc
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(cells)Tj
1.9218 0 Td
(per)Tj
1.5307 0 Td
(ml)Tj
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(for)Tj
/F9 1 Tf
1.3833 0 Td
[()]TJ
/F8 1 Tf
(.)Tj
/F9 1 Tf
0.9015 0 Td
[()7()2()20()-5()-13(\n)-4()10(\n)4()6()]TJ
/F8 1 Tf
(.)Tj
/F9 1 Tf
4.1385 0 Td
[()]TJ
/F8 1 Tf
(.)Tj
/F9 1 Tf
1.0658 0 Td
[()20()2(\n)12()10(\r)6()16()20()14( )7(\n)12(\n)]TJ
/F8 1 Tf
4.4504 0 Td
(and)Tj
1.7291 0 Td
(S2)Tj
1.1736 0 Td
(cell)Tj
1.559 0 Td
(density)Tj
3.1011 0 Td
(in)Tj
1.0375 0 Td
(a)Tj
0.6519 0 Td
(given)Tj
2.3811 0 Td
(culture)Tj
3.0387 0 Td
(were)Tj
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4.8471 0 Td
(directly)Tj
3.2371 0 Td
(using)Tj
2.3868 0 Td
(a)Tj
0.6519 0 Td
(hemocytometer.)Tj
6.7465 0 Td
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1.3946 0 Td
(living)Tj
2.4718 0 Td
(cell)Tj
1.559 0 Td
(types)Tj
2.2734 0 Td
(were)Tj
2.1203 0 Td
(in)Tj
1.0375 0 Td
(the)Tj
1.4683 0 Td
(log)Tj
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(phase)Tj
2.4831 0 Td
(of)Tj
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(growth)Tj
3.0727 0 Td
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(in)Tj
1.0375 0 Td
(bioassays.)Tj
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(bacteria)Tj
3.3789 0 Td
(and)Tj
1.7291 0 Td
(yeast)Tj
2.1883 0 Td
(were)Tj
2.126 0 Td
(pelleted)Tj
3.3278 0 Td
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(2000)Tj
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(x)Tj
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0.686 0 Td
[()]TJ
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0.6349 0 Td
(and)Tj
1.7291 0 Td
(resus-)Tj
-34.3385 -1.2983 Td
(pended)Tj
3.1861 0 Td
(in)Tj
1.0374 0 Td
(sterile)Tj
2.6078 0 Td
(water)Tj
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(to)Tj
1.0262 0 Td
(a)Tj
0.6519 0 Td
(specific)Tj
3.1918 0 Td
(density)Tj
3.1011 0 Td
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3.9061 0 Td
(\(see)Tj
1.7631 0 Td
(below\).)Tj
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2.8857 0 Td
[()]TJ
/F8 1 Tf
(.)Tj
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0.9978 0 Td
[()7()10()12(\n)]TJ
/F8 1 Tf
-35.257 -1.2983 Td
(or)Tj
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1.0941 0 Td
[()]TJ
/F8 1 Tf
(.)Tj
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0.9014 0 Td
[()7()2()20()-5()-13(\n)-4()10(\n)4()6()]TJ
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3.9117 0 Td
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1.9219 0 Td
(resuspended)Tj
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(either)Tj
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(heat)Tj
1.8992 0 Td
(killed)Tj
2.4321 0 Td
(directly)Tj
3.2315 0 Td
(by)Tj
1.1792 0 Td
(autoclaving)Tj
4.8245 0 Td
(or)Tj
-34.7524 -1.2982 Td
(lysed)Tj
2.2336 0 Td
(on)Tj
1.2755 0 Td
(ice)Tj
1.3323 0 Td
(by)Tj
1.1792 0 Td
(sonication)Tj
4.3823 0 Td
(using)Tj
2.3925 0 Td
(a)Tj
0.6519 0 Td
(Fisher)Tj
2.6986 0 Td
(model)Tj
2.7439 0 Td
(100)Tj
1.6498 0 Td
(sonicator)Tj
3.9344 0 Td
(\(40%)Tj
2.2677 0 Td
(amplitude)Tj
4.286 0 Td
(using)Tj
2.3868 0 Td
(four)Tj
1.9162 0 Td
(5)Tj
0.6916 0 Td
(sec)Tj
-36.0223 -1.3039 Td
(pulses\))Tj
2.9877 0 Td
(before)Tj
2.7268 0 Td
(autoclaving.)Tj
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4.7395 0 Td
(cells)Tj
1.9106 0 Td
(and)Tj
1.7178 0 Td
(lysates)Tj
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(were)Tj
2.1147 0 Td
(then)Tj
2.0069 0 Td
(either)Tj
2.5228 0 Td
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2.0466 0 Td
(directly)Tj
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(or)Tj
1.0885 0 Td
(imme-)Tj
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(diately)Tj
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(aliquoted)Tj
3.9627 0 Td
(and)Tj
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(stored)Tj
2.7099 0 Td
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(-80)Tj
1.9105 0 Td
(C)Tj
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2.3925 0 Td
[()]TJ
/F8 1 Tf
(.)Tj
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0.9921 0 Td
[()7()10()12(\n)]TJ
/F8 1 Tf
1.559 0 Td
(or)Tj
/F9 1 Tf
1.0999 0 Td
[()]TJ
/F8 1 Tf
(.)Tj
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0.9014 0 Td
[()7()2()20()-5()-13(\n)-4()10(\n)4()6()]TJ
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3.9118 0 Td
(were)Tj
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(con-)Tj
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(ducted)Tj
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(in)Tj
1.0373 0 Td
(water.)Tj
2.6533 0 Td
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2.3924 0 Td
[()]TJ
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(.)Tj
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1.0658 0 Td
[()20()2(\n)12()10(\r)6()16()20()14( )7(\n)12(\n)]TJ
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4.4504 0 Td
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2.126 0 Td
(conducted)Tj
4.4276 0 Td
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1.0318 0 Td
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(assays)Tj
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(cells)Tj
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(conducted)Tj
4.4277 0 Td
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1.0375 0 Td
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(medium.)Tj
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3.8324 0 Td
[()]TJ
/F8 1 Tf
(.)Tj
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(cells)Tj
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(medium)Tj
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1.1848 0 Td
(incubation)Tj
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(1)Tj
0.6917 0 Td
(h)Tj
0.7483 0 Td
(at)Tj
0.9525 0 Td
(70)Tj
1.5647 0 Td
(C.)Tj
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Q
q
1 j
1 J
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11.9999 0 0 11.9999 200.0125 308.0125 cm
BT
/F14 1 Tf
1 TL
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4.2189 0 Td
(diets)Tj
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Q
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1 j
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(1:1:1)Tj
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-33.3749 -1.2982 Td
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3.7643 0 Td
(5001\),)Tj
2.6873 0 Td
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(\(Fisher\))Tj
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2.6022 0 Td
(yeast)Tj
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(\(US)Tj
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(tropi-)Tj
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(\(Tetra\),)Tj
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12.444 0 Td
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13.9293 0 Td
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12.0982 0 Td
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11.5199 0 Td
(cCold_xviews-_-prodRecCold1)Tj
12.529 0 Td
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2.0863 0 Td
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1.3266 0 Td
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4.5127 0 Td
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(previously)Tj
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(5)Tj
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(liver)Tj
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(8)Tj
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(x)Tj
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(10)Tj
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Q
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1 j
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7.4999 0 0 6.6659 534.7842 139.4645 cm
BT
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2.982 0 Td
(a)Tj
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(drop)Tj
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(of)Tj
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2.4264 0 Td
(from)Tj
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(instar\))Tj
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(under)Tj
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(non-sterile)Tj
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(larvae.)Tj
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(sterile)Tj
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(and)Tj
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(and)Tj
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(days)Tj
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(4)Tj
0.6973 0 Td
(and)Tj
1.7291 0 Td
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(.)Tj
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[()30()]TJ
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(.)Tj
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1.4854 0 Td
[()6()9()-21()-13()12( )7(\n)]TJ
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(monitored)Tj
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0.9411 0 Td
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(32)Tj
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[()]TJ
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(.)Tj
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0.9921 0 Td
[()7()10()12(\n)]TJ
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1.559 0 Td
(in)Tj
1.0375 0 Td
(the)Tj
1.4683 0 Td
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[()6()9()-21()-13()12( )7(\n)]TJ
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1 TL
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q
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1.0261 0 Td
(1)Tj
0.6973 0 Td
(x)Tj
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1 TL
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(.)Tj
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1.5307 0 Td
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(molting,)Tj
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BT
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1 TL
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(assess)Tj
2.5228 0 Td
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[()6()9()-21()-13()12( )7(\n)]TJ
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[()7()10()12(\n)]TJ
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(1)Tj
0.6917 0 Td
(x)Tj
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(10)Tj
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q
1 j
1 J
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7.4999 0 0 6.6659 374.9669 256.422 cm
BT
/F8 1 Tf
1 TL
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ET
Q
q
1 j
1 J
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BT
/F8 1 Tf
1 TL
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(1)Tj
1.2132 0 Td
(x)Tj
0.686 0 Td
(10)Tj
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q
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1 J
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BT
/F8 1 Tf
1 TL
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(8)Tj
ET
Q
q
1 j
1 J
0 w
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BT
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2.4661 0 Td
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(chow)Tj
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(in)Tj
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(each)Tj
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(determine)Tj
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(uals)Tj
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(that)Tj
1.7802 0 Td
(molted)Tj
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1.0261 0 Td
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(second)Tj
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(instar.)Tj
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(12)Tj
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(h)Tj
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(post-inocula-)Tj
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(in)Tj
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(inhibitor)Tj
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(cocktail)Tj
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(previously)Tj
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(described)Tj
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(25)Tj
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1.0148 0 Td
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(\(ThermoFisher\).)Tj
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(Axenic)Tj
3.0274 0 Td
(larvae)Tj
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(processed)Tj
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(0)Tj
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3.7701 0 Td
(buffer)Tj
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7.0015 0 Td
(\(10)Tj
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1.5137 0 Td
[()]TJ
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(M\))Tj
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(fol-)Tj
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(electrophoreses)Tj
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(\(30)Tj
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1.5136 0 Td
[()]TJ
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5.261 0 Td
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1.7121 0 Td
(\(Biorad\))Tj
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(and)Tj
1.7292 0 Td
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3.9175 0 Td
(difluoride)Tj
4.1726 0 Td
(\(PVDF\))Tj
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(\(ThermoFisher\).)Tj
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2.2904 0 Td
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(5%)Tj
1.457 0 Td
(non-fat)Tj
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(milk)Tj
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(PBS)Tj
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(20)Tj
1.1679 0 Td
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(1)Tj
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(h,)Tj
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()Tj
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3.7644 0 Td
(\(1:5000\))Tj
3.5036 0 Td
([)Tj
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(25)Tj
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(or)Tj
-36.0337 -1.2982 Td
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4.0876 0 Td
(\(1:1000,)Tj
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(A2103)Tj
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7.3814 0 Td
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1.8991 0 Td
(goat)Tj
1.9276 0 Td
(anti-rabbit)Tj
4.5014 0 Td
(secondary)Tj
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3.8835 0 Td
(1:5000\))Tj
3.1634 0 Td
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1.1792 0 Td
(visualization)Tj
5.2498 0 Td
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(a)Tj
0.652 0 Td
(chemiluminescent)Tj
7.574 0 Td
(substrate)Tj
3.7984 0 Td
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Q
q
1 j
1 J
0 w
576 737.1 m
36 737.1 l
36 737.6 l
576 737.6 l
f*
36 741.2598 183.2882 23.6976 re
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183.2315 0 0 23.6409 36 741.3165 cm
q
/I0 Do
Q
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Q
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1 J
0 w
7.9999 0 0 7.9999 470.1543 745.7952 cm
BT
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4.2094 0 Td
(and)Tj
1.8567 0 Td
(mosquito)Tj
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576 48.0002 l
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317.7071 34.9228 l
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7.9999 0 0 7.9999 36 36 cm
BT
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1 TL
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(PLOS)Tj
2.8559 0 Td
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4.6417 0 Td
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3.7135 0 Td
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3.5433 0 Td
(s)Tj
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(|)Tj
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(https://doi.or)Tj
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(g/10.137)Tj
3.8197 0 Td
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3.8197 0 Td
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