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Reduced Host Mortality Indicative of Viral In-Fighting?

Johnathan A. Kaiser, Tanya A. Miura, Holly A. Wichman, Christine E. Parent


University of Idaho, Dept. of Biological Sciences & Center for Modeling Complex Interactions

Introduction

Methods
Oral inoculation:
3 day old flies starved for 4 hours and then fed on a viral medium
Viral stocks mixed with a 25% sucrose solution, S2 cell media and red food dye for indication of
viral ingestion:

This study explores the effects of viral single


and co-infection on host mortality by Drosophila C
virus (DCV) and Drosophila X virus (DXV) through
oral inoculation. These are natural viral pathogens
of arthropod species transmitted by feeding and
shown to elicit differing immune responses in host
Drosophila melanogaster.

Fig. 1: D. melanogaster that has ingested viral


medium. Red coloration from food dye is clearly
visible in midgut.

Objective
Determine the effects of single and coinfection (COI) on host mortality by DCV and DXV
through oral inoculation over an extended time
course.

10 replicate vials of 10 flies per treatment (DCV, DXV, COI) and in Mock
Mortality was scored daily as a count of newly deceased flies over the course of several weeks
Flies were transferred to new vials every 7 days.

Results
Fig. 2: Comparison of Total Cumulative Mortality vs Days Post Infection at DPI 20 & 27
A

Days Post Infection (DPI)

Days Post Infection (DPI)

A: Treatments DCV and DXV mortality are significantly greater than Mock (P 0.05)
at DPI 20. Treatment COI (P 0.05). Sharp spike in DXV mortality at DPI 18-20.

B: All treatment (DCV, DXV , COI) mortality counts are significantly greater than
Mock (P 0.05) at DPI 27. Spike in COI mortality at DPI 26-27.

Fig. 3: Comparison of Probability of Death vs Days Post Infection at DPI 20 & DPI 27
B

A: Treatments DCV and DXV have much higher probability of death per
individual than do COI and Mock treatments using counts through DPI 20.

B: Treatments DCV, DXV and COI have higher probability of death per individual than
the Mock treatment. COI probability of death still remains below that of other single
infection treatments using data through DPI 27.

Fig. 4: Statistical Summaries of Mortality in Viral Treatments vs Mock at DPI 20 & 27


DPI 20

DPI 27
Estimate

Std. Error

Z value

Pr ( > |Z| )

COI

0.11912

0.35494

0.993

0.32081

0.35239

0.33224

2.549

0.84693

0.32160

Sex (Male)

-0.26372

0.21514

Estimate

Std. Error

Z value

Pr ( > |Z| )

COI

0.67315

0.25778

2.611

0.009019 **

0.01080 *

0.82505

0.25989

3.175

0.001500 **

2.976

0.00292 **

0.96352

0.25693

3.750

0.000177 ***

-1.226

0.22027

Sex (Male)

0.18661

0.16457

1.134

0.256815

Signifigance codes: 0 *** 0.001 ** 0.01 * 0.05


Treatments DCV and DXV show significantly higher mortality at both DPI 20 and DPI 27. COI does not show significant difference at DPI 20 but does show significant increase at
DPI 27. Sex does not show any significant effect on mortality at either time point.

Discussion

Future Directions

References & Acknowledgments

Levels of mortality at DPI 20 show an antagonistic


effect of DCV and DXV in co-infection
Sharp spike in COI mortality at days 26-27 highly
resemble spike in DXV mortality at days 19-20
DXV appears to require longer time span to induce fatal
infection in host flies than DCV; in Co-infection this lag
appears further extended before inducing host death
Effects may be explained by viral competition for
resources within the host

Confirm rates of infection relative to mortality time


course of viral infections through assays for viral
presence
Differential gene expression studies across treatments
implementing RNA-seq techniques on host and
pathogen transcriptomes to uncover mechanisms of
viral co-infection and innate immune response
Determine rates of transmission between adult flies by
live and deceased infected donors

- Xu J, Cherry S (2014) Viruses and antiviral immunity in


Drosophila. Dev Comp Immunol.
- DaPalma T et al. (2010) A systematic approach to virus
virus interactions. Virus Res.
- Gomariz-Zilber E, Poras M (1995) Drosophila C virus:
experimental study of infectious yields and underlying
pathology in Drosophila melanogaster laboratory populations.
J Inv Pathol.
Thanks to: Ashley DeAguero, Andrea Gonzlez, Luann Scott
& the rest of the Parent Lab

Contact: johnathan.a.kaiser@gmail.com
Project made possible by the CMCI at U Idaho and NIHfunded COBRE grant number: P20GM104420