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Batrachochytrium dendrobatidis

Batrachochytrium dendrobatidis

Bd

Janthinobacterium lividum

Pseudomonas fluorescens,

Bd in vitro. Bd

Lithobates sphenocephalus

J. lividum P.fluorescens.

, Bd-

J. lividum P. fluorescens. Bd

J. lividum

P. fluorescens Bd- . sphenocephalus

Bd

L. sphenocephalus BdBd

Bd

BdBd

Bd- . sphenocephalusBd

BdBd

Bd

L. sphenocephalus L. sphenocephalusJanthinobacterium lividum Pseudomonas fluorescens

L. sphenocephalus L. sphenocephalus

L. sphenocephalus BdBd

L. sphenocephalus Bd BdJ. lividum P. fluorescens L. sphenocephalus

L. sphenocephalus Bd L. sphenocephalusJ. lividum P. fluorescens Bd

Chapter 1: Exposure of Lithobates sphenocephalus to probiotic

bacteria for colonization

Introduction

Conceptual Framework

Batrachochytrium

Bd

,

Bd

Bd

Bd

et al.

Bd

et al.

Bd Bd,

Bd- Bd

Bd

Bd

Bd

Plethodon

cinereus, Bd

Janthinobacterium lividum Pseudomonas fluorescens,

Bd .

Bd Bd

Bd

J. lividum P. fluorescens

et al. et al. ,

Bd

Bd

Lithobates

sphenocephalus Bd

Bd

Bd

Bd

Literature Review

Bd

Bd-

Bd

J. lividum P. fluorescens L. sphenocephalus

Bd,

. J. lividum

P. cinereus Bd

Bd

Bd

Bd

Bd Bd

P. fluorescens J. lividum, Bd

Rana muscosa Rana muscosa

in vivo,

Bd J.

lividum P. fluorescen

Bd .

L. sphenocephalus

J. lividum P. fluorescens

. .

Bd

P. fluorescens J. lividum Bd

P. fluorescens

Rhizoctonia solani

Bacillus harveyi

Vibrio

L.

sphenocephalus, J. lividum P. fluorescens

.

in vivo

Bd

L. sphenocephalus J. lividum P. fluorescens

J. lividum Rana

muscosa Bd

Bd.

bd

L. sphenocephalus Bd

L. sphenocephalus

Bd L.

sphenocephalus

J. lividum P. fluorescens L.

sphenocephalus

J. lividum P. fluorescens Bd

L. sphenocephalus J. lividum P.

fluorescens

L. sphenocephalus

Hypotheses

J. lividum P. fluorescens L. sphenocephalus

J. lividum P. fluorescens L. sphenocephalus L.

sphenocephalus

L. sphenocephalus J.

lividum P. fluorescens

Methodology

Animal Husbandry

L. sphenocephalus

Bd

L. sphenocephalus

Probiotic Cultures

J. lividum P. fluorescens

, ,

.

J. lividum P. fluorescens

J. lividum P.

fluorescens L. sphenocephalus

J. lividum

P. fluorescens

L. sphenocephalus

AMP Collection, Purification, and Efficacy

L. sphenocephalus

L.

sphenocephalus

L. sphenocephalus

J. lividum

P. fluorescens

.

Bd

L. sphenocephalus J. lividum P. fluorescens

J. lividum P.

fluorescens, .

J. lividum P.

fluorescens L. sphenocephalus.

L. sphenocephalus

Probiotic Exposure and Quantification

,

1) J. lividum+/P. fluorescens+

2) J. lividum+/ P. fluorescens; 3) J.

lividum/P. fluorescens+ 4) J. lividum P. fluorescens

.

Statistical Analyses

N =

Results

L. sphenocephalus J. lividum

P. fluorescens

Comparisons of Bacterial Cell Concentrations over Time

J. lividum F = p

J. lividum

L. sphenocephalus

F p

Figure 1 L. sphenocephalus

L. sphenocephalus

J. lividum P. fluorescens

p p p

L. sphenocephalus

J. lividum P. fluorescens.

J. lividum

P. fluorescens F . p P.

fluorescens L. sphenocephalus J.

lividum

AMP production in response to bioaugmentation

F p

J. lividum P. fluorescens

Figure 2 L. sphenocephalus F

P

L. sphenocephalus

Bd

F p

L. sphenocephalus

p

p

p

in

vitro

Discussion

Findings and Conclusions

L. sphenocephalus

J. lividum P. fluorescen

Bd.

L. sphenocephalus

L. sphenocephalus

J. lividum P. fluorescens,

, ,

J. lividum P. fluorescens L. sphenocephalus

J. lividum Bd

P. fluorescens

Bd

L. sphenocephalus

L. sphenocephalus

L. sphenocephalus

J. lividum P. fluorescens

P. fluorescens L. sphenocephalus,

J. lividum. P. fluorescens Bd

J. lividum,

P. fluorescens

L. sphenocephalus

Bd

P.

fluorescens Bd

P. fluorescens J. lividum,

Bd J. lividum

Bd,

Recommendations for Future Research

J. lividum P. fluorescens

L. sphenocephalus,

Bd.

Bd

Bd

Bd-

in vivo

Bd

Bd

Chapter 2: Susceptibility to Bd after Successful Colonization of

L. sphenocephalus with J. lividum and

P. fluorescens

Introduction

Conceptual Framework

Batrachochytrium dendrobatidis Bd

Bd

Bd

Bd

Bd

Bd

Bd

Xenopus laevis) Bd

Bd .

Bd

Literature Review

Bd

Bd

.

Janthinobacterium lividum, Bd

J. lividum

Bd Bd

J. lividum

Bd, .

Pseudomonas fluorescens Bd in vitro

P.

fluorescens

J. lividum P. fluorescens

Bd

.

Bd

Bd,

Bd

. Bd

Bd

Bd

Bd.

.

Bd

Bd

.

Bd

Bd

. Bd ,

Bd

et al.

Lithobates sphenocephalus;

.

Bd-

Plethodon cinereus, J. lividum

Bd

J. lividum

P.fluorescens, Bd

Lithobates sphenocephalus J. lividum P. fluorescens

,

L. sphenocephalus

J.lividum P.fluorescens.

Bd L.

sphenocephalus

Bd-

J. lividum P. fluorescens Bd ,

L. sphenocephalus Bd.

Hypotheses

J. lividum P. fluorescens L. sphenocephalus

Bd

Bd

Bd

in vitro et al. .

L. sphenocephalus Bd

J. lividum P. fluorescens

L. sphenocephalus

J. lividum P. fluorescens Bd

Methodology

Exposure of L. sphenocephalus to Bd

L. sphenocephalus

Bd

L. sphenocephalus.

Bd L.

sphenocephalus Bd

L.

sphenocephalus.

Bd

Bd

Bd

Bd

Bd Bd

L.

sphenocephalus Bd

Bd

J. lividum P. fluorescens J. lividum P. fluorescens +

J. lividum, P. fluorescens, Bd

Bd

Bd

L. sphenocephalus

Bd

Probiotic and Bd quantification

L. sphenocephalus Bd

J. lividum

P.fluorescens

J. lividum P.

fluorescens

Bd

Bd

L. sphenocephalus Bd-

Bd

AMP Collection, Purification, and Quantification

Bd

L. sphenocephalus

.

L. sphenocephalus

Quantitative Polymerase Chain reaction (qPCR) Analysis

Bd

Bd

Bd

Bd

Bd

Bd

Statistical Analyses

L. sphenocephalus

N

N

Bd

Bd Bd

Bd

Results

Bd-induced Changes of AMP Production

L. sphenocephalus F

= p F = p

Bd F p

.

Bd P. fluorescens/Bd F = p

J.lividum/Bd F p

Bd

L. sphenocephalus J.lividum P. fluorescens .

Bd L.

sphenocephalus

F4,36 p F8, 36 = p

Bacterial Concentrations Fluctuates Post-Bd Infection

F p

L. sphenocephalus

F p

Figure 3

Figure 4. L. sphenocephalus

P

Bd

F p J. lividum F1,7 p P. fluorescens F

p

Probiotic Inhibition of Bd zoospores

Bd

F1,4 p

Bd L. sphenocephalus

J. lividum

Bd

F p

J. lividum P.

fluorescens p L.

sphenocephalus J. lividum Bd

P. fluorescens p Bd

J.lividum p

Bd

Figure 5 L. sphenocephalus

Discussion

Discussion of Findings and Conclusions

Bd

J. lividum P. fluorescens Bd

L. sphenocephalus

L. sphenocephalus

J.lividum P. fluorescens

J. lividum P.

fluorescens

Bd . J. lividum

P.fluorescens

L. sphenocephalus

.

L. sphenocephalus

J.lividum P. fluorescens

L. sphenocephalus

Bd-

J. lividum P. fluorescens,

L. sphenocephalus

Bd

L. sphenocephalus Bd

J. lividum P. fluorescens

L. sphenocephalus J. lividum P.

fluorescens, Bd

L.

sphenocephalus

L. sphenocephalus

.

J. lividum P. fluorescens

Bd- L.

sphenocephalus J. lividum P. fluorescens

K

L.

sphenocephalus

et al. .

J. lividum P. fluorescens

L.

sphenocephalus Bd

Bd, L. sphenocephalus

Bd-

Bd

Bd L. sphenocephalus

Bd

J. lividum P.

fluorescens

J. lividum P. fluorescens

Flavobacterium johnsoniae and P. fluorescens Bd- Alytes

obstetricans

Recommendations for future research

Bd

.

.

Bd

J. lividum P. fluorescens Bd L.

sphenocephalus

. in vivo

L. sphenocephalus,

Bd

Bd

Bd

Bd

.

Bd

Bd

L. sphenocephalus J. lividum P. fluorescens

L. sphenocephalus

J. lividum P. fluorescens.

Bd

Daphnia magna Bd

L. sphenocephalus

D. magna L. sphenocephalus

Bd.

Bd.

J. lividum P. fluorescens

Bd

L. sphenocephalus with J. lividum P. fluorescens

Bd .

Chapter 3: Addition of J. lividum and P. fluorescens to

BD-Infected L. sphenocephalus

Introduction

Conceptual Framework

.

.

Batrachochytrium

dendrobatidis Bd Bd

Bd Bd

Bd

Bd

Bd-

Bd

Bd-

Bd

.

Bd .

. .

Bd

Janthinobacterium lividum

Bd .

Hyla spp.

et al.

Literature review

Bd J. lividum Pseudomonas fluorescens,

in vitro

Bd Bd

J. lividum Bd

P. fluorescens,

Bd

J. lividum

P. fluorescens Bd

Bd

Bd

Bd-

J. lividum P.

fluorescens,

J. lividum P.

fluorescens Bd

Lithobates sphenocephalus in

vitro. L. sphenocephalus Bd

J. lividum P. fluorescens

Bd .

Bd , L. sphenocephalus J. lividum P. fluorescens

Bd

J. lividum P. fluorescens Bd

Bd

Bd J. lividum P. fluorescens Bd

L. sphenocephalus Bd

Hypotheses

J. lividum P. fluorescens L. sphenocephalus

Bd Bd L. sphenocephalus

J. lividum P. fluorescens. Bd

L. sphenocephalus

.

Bd

L. sphenocephalus

Methodology

Bacterial Exposure of Bd-Infected L. sphenocephalus

Acheta domestica

Bd

L. sphenocephalus.

Bd

Bd

Bd

Bd

L.

sphenocephalus Bd

Bd

J. lividum

P. fluorescens J. lividum+/P. fluorescens+ J. lividum+/P. fluorescens-

J. lividum-/P. fluorescens +. Bd

Bd J. lividum P.

fluorescens.

Bd

Bd and probiotic quantification

L. sphenocephalus

Bd

Bd

Bd

J lividum P. fluorescens

J. lividum P.

fluorescens

J.

lividum P. fluorescens

AMP collection, purification and quantification

Bd

L. sphenocephalus

.

Statistical Analyses

L. sphenocephalus

N

N

Bd

Results

Bd infection effects on bacterial colonization

J. lividum P. fluorescens L. sphenocephalus

Bd

L.

sphenocephalus J.lividum P. fluorescens.

Bd .

J. lividum P. fluorescens

J. lividum P. fluorescens N

F

p

AMP release during Bd infection and subsequent bacterial exposure

Bd F3,22 p

F3,22 p

F p

F3,22 p Bd

L. sphenocephalus

F p

Bd zoospore loads in response to probiotic intervention

Bd- Bd

L. sphenocephalus

J. lividum P. fluorescens Bd

F p

Bd

Bd J. lividum P. fluorescens

Bd-

J. lividum P. fluorescens L. sphenocephalus

Bd

F p

F p Bd

Bd

Discussion

Discussion of findings, conclusions

Bd- L.

sphenocephalus, J. lividum P. fluorescens

. L. sphenocephalus

J.lividum P.

fluorescens Bd-

.

L. sphenocephalus

L.

sphenocephalus

Bd

Bd

J. lividum P. fluorescens

L. sphenocephalus Bd

Bd L. sphenocephalus

J. lividum P.

fluorescens

.

L. sphenocephalus

J. lividum P. fluorescens.

. Bd

Bd L. sphenocephalus

Bd

Bd

L.

sphenocephalus Bd

J.lividum

P.fluorescens

Bd, Bd

Bd

L. sphenocephalus Bd

et al. et al . Lithobates clamitans

Bd ,

L. sphenocephalus

J. lividum P.

fluorescens L. sphenocephalus Bd

Bd

L. sphenocephalus Bd

Bd

J. lividum P. fluorescens Bd

Bd

Bd,

Bd

Bd,

Bd Bd

Recommendations for future research

Bd Bd

.

Bd Bd

in vitro

.

Bd

Bd

Bd

Bd

bd

Bd-

L. sphenocephalus Bd .

J. lividum P. fluorescens

.

Bd

Bd

Bd

J. lividum P.fluorescens

Staphylococcus spp.,

Bd-

L. sphenocephalus

J. lividum

P.fluorescens

J. lividum P. fluorescens Bd

J. lividum P. fluorescens

Bd L. sphenocephalus

L. sphenocephalus Bd

Bd

Bd

L. sphenocephalus Bd

.

Bd-

Bd-

Pseudomonas fluorescens Journal of Bacteriology

Batrachochytriumdendrobatidis Molecular Ecology 22

Applied andEnvironmental Microbiology, 75(21), 6635-6638.

PLoS One 5

Ecohealth 8

Frontiers in Ecology and the Environment 5.10,

The role of cutaneous bacteria in resistance of Australian tropicalrainforest frogs to the amphibian chytrid fungus Batrachochytriumdendrobatidis

Gesunde Pflanzen 59

Diseases of AquaticOrganisms 68

Diseases of Aquatic Organisms 68

Ecology Letters.

Hyla cinereaBatrachochytrium dendrobatidis PloS One 7

Batrachochytrium dendrobatidisPloS one 8

Ecology, 86,

Plethodoncinereus. Journal of Chemical Ecology

Janthinobacterium lividumPlethodon cinereus Journal of Chemical Ecology 34

Scientificreports

Batrachochytrium dendrobatidisFungal Genetics and

Biology, 49,

Disease ecology: community structure andpathogen dynamics

General andComparative Endocrinology 176

Proceedings of the National Academy of Sciences 107

Herpetologica, 63,

Environmental Science & Technology 41

Molecular Biology and Evolution 19

Biotechnology andApplied Biochemistry 48

Rana muscosa Copeia 2001

Journal of Zoo andWildlife Medicine 39

Conservation Biology 26

Diseases of Aquatic Organisms 72

Batrachochytrium dendrobatidisOikos 118

Pseudomonas fluorescens

Saccharomyces cerevisiae Antonie van Leeuwenhoek 97

Batrachochytrium dendrobatidis Xenopus laevis Ecotoxicologyand Environmental Safety 71

Nature Reviews Microbiology 10

Freshwater Biology 57

Diseases of Aquatic Organisms 83

The ISME Journal 3

Food Control, 32

Archives of Environmental Health: AnInternational Journal 46

Proceedings of the National Academy of Sciences

Batrachochytrium dendrobatidisEmerging Infectious

Diseases 9

Functional Ecology 22

Pseudomonas fluorescens Applied and EnvironmentalMicrobiology 76

Batrachochytrium dendrobatidisDiseases of Aquatic Organisms 77

Lactobacillusrhamnosus Lactobacillus reuteri

Candida albicans. Infectious Diseases in Obstetrics andGynecology 2012

PloS one 9

y.Journal of Archaeological Method and Theory 17 4

Batrachochytrium dendrobatidisRana muscosa Biological

Conservation, 143

Batrachochytriumdendrobatidis PloS one 8

Copeia

Microbial Ecology 2

Lithobatescatesbeianus Mycopathologia

Microbiology: Laboratory theory andapplication

PLoS biology 6

Frontiers inMicrobiology 5

The ISME journal 8

Gastroenterology, 121(3),

Applied and Environmental Microbiology 70

The ISME Journal 6

Journal of Hygiene, 38

Annual Reviews in Microbiology,3,

Aquaculture 164

Batrachochytrium dendrobatidis BiologicalConservation 152

Batrachochytrium dendrobatidisRana muscosa Journal of

Chemical Ecology 38

Nature Protocols 1

Niche construction: theneglected process in evolution

Janthinobacterium lividum Journal of Applied Microbiology 102

Ecology 85

Rana pipiensGlobal Change Biology 18

The Journal ofExperimental Biology 216

Batrachochytrium dendrobatidis

Journal of Wildlife Diseases 43

Batrachochytrium dendrobatidisDiseases of Aquatic Organisms 75

Bioscience 59

Microbiology andMolecular Biology Reviews 74

PLoS genetics 9

Immunological reviews 166

Batrachochytrium dendrobatidisDevelopmental & Comparative

Immunology 26

Developmental & Comparative Immunology 29

Rana muscosa Developmental &Comparative Immunology 30

Biochimica et Biophysica Acta (BBA)-Biomembranes 1788

Proceedings of the National Academy ofSciences 105

Batrachochytrium dendrobatidis Diseases ofAquatic Organisms 82

Journal ofHerpetology

Proceedings of the National Academy of Sciences,108(40), .

Aquaculture 256

Ambystoma tigrinumJournal of Wildlife Diseases 44

Fusarium oxysporum Aspergillus Pseudomonas fluorescensJournal of Pharmacy and

Biological Science 1

The ecology of the nitrogen cycle

Rana pipiensDevelopmental & Comparative

Immunology 33

Environmental Toxicology and Chemistry, 16(2),

Oecologia 93

Journal of Parasitology 96

Diseases of AquaticOrganisms 77

Batrachochytrium dendrobatidisMicrobes and Infection 13

Batrachochytrium dendrobatidisFungal Ecology 4

Host-pathogen dynamics of amphibian chytridiomycosis: the role of the skin microbiome in health anddisease.

Biotropica 43

The ISME journal

Batrachochytrium dendrobatidis Diseases of AquaticOrganisms, 77

Rana

Diseases of aquatic organisms 98

PloS one 9