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Research ArticleSynergistic Effects of Salinity and Temperature onthe Survival of Two Nonnative Bivalve Molluscs Perna viridis(Linnaeus 1758) and Mytella charruana (drsquoOrbigny 1846)

Wei S Yuan Linda J Walters Sasha A BrodskyKimberly R Schneider and Eric A Hoffman

Department of Biology University of Central Florida 4000 University Boulevard Orlando FL 32816-2368 USA

Correspondence should be addressed to Linda J Walters lindawaltersucfedu

Received 29 April 2016 Accepted 26 June 2016

Academic Editor Baruch Rinkevich

Copyright copy 2016 Wei S Yuan et al This is an open access article distributed under the Creative Commons Attribution Licensewhich permits unrestricted use distribution and reproduction in any medium provided the original work is properly cited

This study examined the combined salinity and temperature tolerance of two marine bivalve molluscs Perna viridis and Mytellacharruana which have recently invaded the southeastern United States It is essential to understand the role that these abioticvariables play in invasions and establishment of nonnative species We simultaneously explored survival at three salinity ranges (5ndash9 20ndash225 and 35ndash40 ppt) in both cold and warm water for juveniles and adults of both species We determined that Perna viridiscan survive at a wide range of temperatures (9ndash35∘C) when the salinity is 35ndash37 ppt however as salinity decreased the thermalsurvival range for P viridis became narrower WithM charruana our data suggest that juvenile and adult individuals can surviveat a wide range of salinities (5ndash40 ppt) at 20∘C but the salinity tolerance range narrowed as the temperature decreased or increasedAdditionally we observed that temperature rapidly impacted survival of P viridis andM charruana (within hours) while salinityimpacts were more gradual (days to weeks) These data can be used to help predict successful introductions and future expansionsof P viridis andM charruana in introduced habitats

1 Introduction

Global transportation has provided a means for marineinvertebrate species to be transferred to new locations [1ndash3] Species can be transported as larvae in ballast waterand as adults attached to ship hulls through the aquariumtrade recreational boating and aquaculture (eg [1 4ndash8])Globally molluscs are infamous invaders in both fresh andsaltwater as they are easy to transport difficult to identifyand difficult to eradicate once established [1 9ndash12] As a resultthe number of invasive molluscs continues to increase [13]For example more than 88 species of molluscs have becomeestablished in theUnited States via ballast water and hull foul-ing [13] Although modes of global dispersal are becomingbetter known formolluscs less remains understood about thefactors that affect establishment of these organisms in newlocations

Perna viridis and Mytella charruana are two speciesof nonnative marine molluscs that have recently invaded

Florida (USA) and are potential causes of concern along thesoutheastern United States Both P viridis andM charruanahave been documented on intertidal oyster reefs in Florida[14 15] and have been found to negatively impact theeastern oyster Crassostrea virginica [12] We are interested inunderstanding how the abiotic conditions of temperature andsalinity independently and in synergy influence the survival(and subsequent establishment) of these two nonnative mol-luscs and speculate how these factors may further restrictdistributions in estuaries along the eastern Atlantic Oceanfrom Florida to Nova Scotia (Canada) where the easternoyster is located [16]

There is a wealth of information on the synergistic effectsof temperature and salinity on native molluscs [17ndash19] whileother researchers have examined the combined effects ofthese abiotic factors to better understand invasion potentialofmolluscs around the globe (eg [20 21]) For example Saraet al [21] investigated the invasive Brachidontes pharaonisnow found in the Mediterranean Sea by simultaneously

Hindawi Publishing CorporationJournal of Marine BiologyVolume 2016 Article ID 9261309 14 pageshttpdxdoiorg10115520169261309

2 Journal of Marine Biology

testing a range of salinities and temperatures to determine itsphysiological tolerance for feeding These researchers deter-mined that the clearance rate of B pharaonis declined underconditions of low salinity and low temperature Likewise Hiset al [20] examined the combined effects of temperature andsalinity on the invasive Japanese oyster C gigas in France anddetermined that the oyster exhibited the best growth at highertemperatures

Previous studies have independently investigated the roleof salinity and temperature in survival of P viridis and therole of salinity in survival in M charruana [22ndash24] Pernaviridis has been documented in salinities ranging from 19 to44 ppt in its native habitat [25] Segnini de Bravo et al [22]determined that P viridis could tolerate an even wider rangeof salinities under laboratory conditions (25∘C) and the lowerand upper lethal salinities were 0 and 64 ppt respectivelyPerna viridis survived at temperatures ranging from 10 to42∘C in the laboratory [25ndash29]This broad temperature rangeshould enable P viridis to survive in both tropical andtemperate zones

Mytella charruana survived in salinities ranging from2 to 40 ppt at 20∘C [24] Not much is known about thethermal tolerance for M charruana so the first goal of ourstudy was to address how temperature influenced survivalin M charruana independent of salinity By addressing thisknowledge gap we were then able to determine appropriatetemperature ranges for our salinity-temperature interactionexperiments

The overall objective of this study was to investigate thesimultaneous effects of salinity and temperature on nonnativePerna viridis and Mytella charruana to better understandtheir respective invasion potentials Specifically we addressedthe following questions (1)What is the temperature tolerancerange for survival of Mytella charruana (2) How do salinityand temperature interact to influence the survival of adultand juvenile Perna viridis (3) How does the interactionof salinity and temperature influence the survival of adultand juvenile Mytella charruana These results will aid inpredicting suitable environments where adult and juvenileindividuals of both molluscs may become established andpredict future range expansions

2 Materials and Methods

21 Study Species Perna viridis the Asian green mussel isnative to the Indo-Pacific region along the Southeast Asiancoastline from the Arabian Gulf to southern China andsouthern Japan [30] and was first documented in the UnitedStates in 1999 in the cooling system of an electrical powerplant in Tampa Florida [31] Currently P viridis is wellestablished in the United States and extends from the TenThousand Islands to the northern Gulf of Mexico on the westcoast of Florida and along the eastern United States coastlinefrom Stuart FL (27∘ 098311015840N 80∘ 116041015840W) to northernGeorgia (32∘ 012061015840N 80∘ 595691015840W) [15 32 33] In TampaBay P viridis had the highest density of all observed locationswithin the United States with 2505 individuals mminus2 [34]

Mytella charruana commonly known as the charru mus-sel or sururu is native to the Galapagos Islands the Pacific

coast from Mexico to Ecuador [35ndash38] and the Atlanticcoast from Argentina to Venezuela [39] Like P viridis thediscovery ofM charruana in the United States occurred at apower plantMytella charruana was reported in JacksonvilleFL in 1987 it was considered to be locally extirpated by spring1988 [39] In 2004 however M charruana was found inMosquito Lagoon 212 km south of Jacksonville [14] Mytellacharruana has now been recorded along the southeasternUnited States coast from Titusville FL (28∘ 372211015840N 80∘483711015840W) to Charleston SC (32∘ 464521015840N 79∘ 575391015840W)[15] Densities of this nonnative species in the United Stateshave been measured up to sim12 individuals mminus2 [15] Thisdensity is much lower than that in their native habitats whereM charruana densities as high as 11036 mussels mminus2 havebeen reported [40]

22 Thermal Tolerance of Mytella charruana Mytella char-ruana (20ndash54mm) were collected in cold and warm monthsbetweenDecember 2007 andOctober 2009 from two floatingdock sites (Arlington Lions Club and Lonnie Wurn) inJacksonville FL (Table 1(a) Figure 1)The location of samplescollected for our experiments varied because we could notcollect the 300ndash400 mussels needed per experiment from asingle site Stenyakina et al [41] found thatM charruanaweresexually mature at 125mm For thermal tolerance trials weused adult mussels that were ge20mm

Individuals were acclimated to the laboratory environ-ment at 20∘C for 13 days in a recirculating aquaria (151 L)with water from the collection site This temperature waschosen as an intermediate for acclimation of all individualsas we previously determined that this species placed inwater at this temperature always survived in high numbersregardless of whether it represented a temperature increase ordecrease from field conditions In Florida between 1981 and2010 the temperature ranged from 3 to 26∘C during January(coldest month) and from 19 to 35∘C in July (warmest month)[42] Thirteen days was used as the acclimation period asit represented sufficient time for any mortality associatedwith collection to occur Prior to running these trials wedocumented that individuals that survived at least 13 dayswould survive for months in 20∘C waters (Yuan pers obs)Additional details of each trial are listed in Table 1(a)

After acclimation groups of 25 adult individuals rangingin length from 20 to 54mm were placed in separate aeratedaquaria (21 L) that contained 14 L of water in a ratio of 25collection site water 75 artificial seawater (Instant Oceansalts and deionized water) that maintained the salinity fromthe collection site It was not practical to collect sufficientnatural seawater for all trials Survival in preliminary trialswas 100 with this ratio of seawater to natural seawater(Yuan pers obs) We divided the range of experimentaltemperatures into two seasons cold for winter and warm forsummer conditions To avoid shocking the M charruanacold and warm experiments were correlated with field col-lection temperatures

Cold experiment A was conducted in a walk-in freezer at6∘C at the University of Central Florida at 6 9 11 and 13∘CWarm experiment B examined survival at 20 31 and 36∘Cwhile experiment C investigated survival over a narrower

Journal of Marine Biology 3

Table1(a)S

ummaryo

fMytellacharruanathermalexperim

ents

(b)S

ummaryo

fPerna

virid

isand(c)M

ytellacharruanatemperature-salinity

interactionexperim

ents

Acclimationperio

dwasthen

umbero

fdaysinho

ldingtanks

priortos

tartofexperim

ents

Adjustm

entperiodwasthen

umbero

fdaystotransition

fromho

ldingtanktem

perature(20∘C)

totre

atmenttem

peratures

Experim

entp

eriodwas

then

umbero

fdaystria

lran

inclu

ding

adjustmentp

eriodCa

pitalletterscorrespo

ndto

thee

xperim

entaltreatmentspresentedin

Figures2

3and

5

(a)Mytellacharruana

Experim

ents

Collectionsites

Latitud

eamplong

itude

Collection

temperatures

(∘ C)

Mussel

size(mm)

Collection

salin

ity(ppt)

Acclimation

perio

d(days)

Adjustm

ent

perio

d(days)

Experim

ent

perio

d(days)

Musselsper

tank

replicate

tank

s

Temperatures

teste

d(∘ C

)

Salin

ities

teste

d(ppt)

Adult

Arling

tonLion

sClub

3037

82721015840

N13

20ndash4

610

1315

33254

6911and

13NA

Acold

JacksonvilleFL

816208281015840W

Adult

Lonn

ieWurn

3037

57981015840

N28

22ndash54

813

1533

253

2031and36

NA

Bwarm

JacksonvilleFL

815851651015840W

Adult

Lonn

ieWurn

3037

57981015840

N27

25ndash4

610

1315

33254

202328and

31NA

Cwarm

JacksonvilleFL

8152

51651015840

W

(b)Pernavirid

is

Experim

ents

Collectionsites

Latitud

eamplong

itude

Collection

temperatures

(∘ C)

Mussel

size(mm)

Collection

salin

ity(ppt)

Acclimation

perio

d(days)

Adjustm

ent

perio

d(days)

Experim

ent

perio

d(days)

Musselsper

tank

replicate

tank

s

Temperatures

teste

d(∘ C

)

Salin

ities

teste

d(ppt)

Juvenile

Hugueno

tPark

304052371015840N

1012ndash38

365

1528

103

69and20

52036

Acold

JacksonvilleFL

81403211015840

WJuvenile

Hugueno

tPark

304052371015840N

1012ndash38

365

1528

103

2031and35

52036

Bwarm

JacksonvilleFL

81403211015840

WAd

ult

Mun

icipalMarina

298919731015840N

1858ndash114

355

1528

103

69and20

52035

Ccold

StA

ugustin

eFL

8131

00511015840

WAd

ult

Mun

icipalMarina

298919731015840N

2857ndash113

375

1528

103

2031and35

52037

Dw

arm

StA

ugustin

eFL

8131

00511015840

W

(c)Mytellacharruana

Experim

ents

Collectionsites

Latitud

eamplong

itude

Collection

temperatures

(∘ C)

Mussel

size(mm)

Collection

salin

ity(ppt)

Acclimation

perio

d(days)

Adjustm

ent

perio

d(days)

Experim

ent

perio

d(days)

Musselsper

tank

replicate

tank

s

Temperatures

teste

d(∘ C

)

Salin

ities

teste

d(ppt)

Juvenile

Fire

Station38

3039

00771015840

N17

5ndash12

57

1528

103

69and20

522540

Ecold

JacksonvilleFL

816415761015840W

Juvenile

Fire

Station38

3039

00771015840

N29

4ndash12

97

1528

103

2031and33

522540

Fwarm

JacksonvilleFL

816415761015840W

Adult

Fire

Station38

3039

00771015840

N16

20ndash38

57

1528

103

69and20

522540

Gcold

JacksonvilleFL

816415761015840W

Adult

Arling

tonLion

sClub

3037

82721015840

N28

20ndash31

57

1528

103

2031and33

522540

Hw

arm

JacksonvilleFL

816208281015840W


0 45 9 18

(km)

N

W E

S

0 105 210 420

(km)

Current distribution range

(b)

(a)

Perna viridisMytella charruana

Figure 1 (a) Map of current distribution of Perna viridis andMytella charruana and (b) collection sites for both species along Atlantic coastof Florida 998771 indicates collection sites for P viridis at Huguenot Park (HP) and Municipal Marina (MM) e indicates collection sites for Mcharruana at Fire Station 38 (FS38) Arlington Lions Club (ALC) and Lonnie Wurn (LW)

range of temperatures (20 23 28 and 31∘C) Experimentaltemperatures were controlled by aquarium heaters (Hydor75W Top Fin 25W or Finnex 50W heaters) All aquariamedia started at their collection site salinity (see Table 1)and 20∘C (control) Temperature adjustments occurred every5 days for 15 days in equal increments depending on thedifference between 20∘C and the preplanned experimentaltemperatures (Table 1(a)) Once the treatment temperatureswere reached on day 15 trials continued for 18 addi-tional days Salinities and temperatures were monitored andadjusted as needed twice each day for the duration of eachtrial

There were four replicate aquaria for each temperaturetreatment for experiments A and C (119899 = 4) and threereplicates for experiments B (119899 = 3) based on availability(Table 1(a)) Individuals were fed daily with a 10 algalsolution made from an algal paste containing Chaetoceros-B Phaeodactylum tricornutum and Nannochloropsis oculatafrom Innovation Aquaculture (SPAT formula brand) Eachtank of 25 individuals was fed 25mL (01mL per mussel) thevolume of food per tank was reduced with mussel mortalityMortality was checked daily during each trial Individualswere considered dead if they remained gaping and did notrespond to physical stimulus Dead mussels were removedfrom tanks

23 Perna viridis Interaction of Salinity and Tempera-ture Perna viridis were collected in winter and summermonths between February 2009 and September 2010 alongthe northeast coast of Florida from floating docks in StAugustine FL and a jetty in Jacksonville FL (Table 1(b)Figure 1) These locations were separated by 64 km Pernaviridis were placed in recirculating 210 L tanks in laboratoryfor acclimation using methods described above Shell lengthfor reproductive maturity in this species was 40mm [43] wetherefore categorized P viridis less than 40mm as juvenilesand greater than 40mm as adults Although adult P viridishave been reported to reach over 220mm in length alongthe southeastern Atlantic coastline no mussels greater than120mm were used in our trials Sizes for each trial areprovided in Table 1(b) All were acclimated to aquariumconditions for 5 days prior to the start of each experiment

After acclimation the procedures were similar to theM charruana thermal tolerance trials We used 20∘C as thestarting temperature in both the cold and warm experimentsto facilitate comparisons The starting salinities for theexperiments with P viridis ranged from 35 to 37 ppt depend-ing on collection site salinity (Table 1(b)) Each trial hadnine salinity-temperature treatments which combined threesalinity levels (5ndash9 20 and 35ndash37 ppt) and three temperaturelevels for the cold (6 9 and 20∘C) andwarm (20 31 and 35∘C)


0102030405060708090

100Su

rviv

al o

ver t

ime (

)

(Days)0 3 6 9 12 15 18 21 24 27 30 33

CB

BA

6∘C

9∘C

11∘C

13∘C

(a)

Surv

ival

ove

r tim

e (

)

0102030405060708090

100

(Days)0 3 6 9 12 15 18 21 24 27 30 33

AB

C20∘C

31∘C

36∘C

(b)

Surv

ival

ove

r tim

e (

)

(Days)0 3 6 9 12 15 18 21 24 27 30 33

0102030405060708090

100

AA

BC

20∘C

23∘C

28∘C

31∘C

(c)

Figure 2 Survivorship curves ofMytella charruana for temperature experiments Shell lengths 20ndash54mm Capital letters correspond to theexperimental treatments presented in Table 1(a) (a) Cold experiment (temperatures 6 9 11 and 13∘C) (b) warm experiment (temperatures20 31 and 36∘C) and (c) warm experiment (temperatures 20 23 28 and 31∘C) Upper case letters indicate significant differences insurvivorship (119901 le 005) among temperature treatments Dotted lines show the end of the 15-day temperature adjustment periods

trials Each treatment had three replicate aquaria (119899 = 3) andeach aquarium held 10 individuals Adjustments in salinityand temperature occurred every 5 days during the adjustmentperiod anddepended on the difference between the collectionsite and the preplanned experimental treatments Individualsremained in their experimental tanks throughout each trialThe final salinity-temperature treatments were reached atthe same time for all combinations at 15 days Then eachexperiment ran for an additional 13 days Additional detailsare provided in Table 1(b) Throughout the experiments Pviridis were fed with an algal solution as described above(04mL per adult 02mL per juvenile mussel)

24 Mytella charruana Interaction of Salinity and Tem-perature Mytella charruana were collected in warm andcold months between February 2009 and September 2010on floating docks in Jacksonville (Table 1(c) Figure 1)

Individuals were placed in recirculating aquaria (76ndash151 L)with water from collection sites All samples were held for7 days before experiments began Mytella charruana was fedwith an algal solution of 01mL per mussel throughout theexperiment

The experimental procedures were similar to the P viridisinteraction trials For M charruana the starting salinitiesin the estuaries at the time of collection ranged from 5to 9 (Table 1(c)) Each trial had nine salinity-temperaturetreatments of three salinity levels (5ndash9 225 and 40 ppt) andthree temperature levels for the cold (6 9 and 20∘C) andwarm (20 31 and 33∘C) trials The final salinity-temperaturetreatmentswere reached at the same time for all combinationsat 15 days and then the experiments were run for 13 additionaldays Each aquarium contained 10 individuals and therewere three replicate aquaria for each treatment combinationAdditional details are provided in Table 1(c)


Juve

nile

sur

viva

l ove

r tim

e (

)

0102030405060708090

100

(Days)0 4 8 12 16 20 24 28

ColdPerna viridis

DDCDCD

ABDABA

5 6∘C20 6∘C36 6∘C5 9∘C20 9∘C

36 9∘C5 20∘C20 20∘C36 20∘C

(a)Ju

veni

le s

urvi

val o

ver t

ime (

)

0102030405060708090

100

(Days)0 4 8 12 16 20 24 28

WarmPerna viridis

DACABEFBC

CDFCDEC

5 20∘C20 20∘C36 20∘C5 31∘C20 31∘C

36 31∘C5 35∘C20 35∘C36 35∘C

(b)

(Days)0 4 8 12 16 20 24 28

Adul

t su

rviv

al o

ver t

ime (

)

0102030405060708090

100Cold

Perna viridis

C

D

CD CDCD

CD A

BAB

5 6∘C20 6∘C35 6∘C5 9∘C20 9∘C

35 9∘C5 20∘C20 20∘C35 20∘C

(c)

Adul

t su

rviv

al o

ver t

ime (

)

0102030405060708090

100

(Days)0 4 8 12 16 20 24 28

WarmPerna viridis

CDBC

AA

B

DE

EF

C

5 20∘C20 20∘C37 20∘C5 31∘C20 31∘C

37 31∘C5 35∘C20 35∘C37 35∘C

(d)

Figure 3 Survivorship curves over 28 days for Perna viridis in the temperature-salinity interaction experiments (a) Juvenile cold experiment(temperatures 6 9 and 20∘C salinities 5 20 and 36 ppt) (b) juvenile warm experiment (temperatures 20 31 and 35∘C salinities 520 and 36 ppt) (c) adult cold experiment (temperatures 6 9 and 20∘C salinities 5 20 and 35 ppt) and (d) adult warm experiment(temperatures 20 31 and 35∘C salinities 5 20 and 37 ppt) Upper case letters indicate significant differences of 119901 le 005 among treatmentsusing survivorship analysis Dotted lines show the end of the 15-day temperature adjustment periods

25 Data Analysis Kaplan-Meier survival analysis [44] wasused to compare survivorship curves over the entire lengthof each experiment using pairwise comparisons to determineif significant differences were present among treatments [45]Additionally survival at the end of each trial forM charruana

in the thermal tolerance experiments was analyzed with one-way ANOVA with tank as the experimental unit Similarlythe interaction of salinity and temperature at the end ofeach trial for P viridis and M charruana was analyzed withtwo-way ANOVA Both tests were run using JMP v10 [44]


Table 2 One-way ANOVA on the final survival for Mytellacharruana thermal tolerance experiments

Experiments Source DF MS 119865 119901

Cold A 6 9 11 amp 13∘C 3 6156 398 00352lowastError 12 1548

Warm B 20 31 amp 36∘C 2 48909 25153 lt00001lowastError 9 194

Warm C 20 23 28 amp 31∘C 3 37156 3518 lt00001lowastError 12 1056

lowastSignificantly different at 119901 lt 005 level

Homogeneity of variances was checked using Levenersquos testand normality was checked using Shapiro-Wilk test trans-formations of data were performed as required Tukeyrsquos aposteriori tests were used to determine significant differencesamong treatments

3 Results

31 Mytella charruana Temperature Tolerance ExperimentsCold experiment A examined survival at 6 9 11 and13∘C treatments were significantly different based uponsurvivorship analysis (119901 lt 0001 Figure 2(a)) The 13∘Ctreatment had a steady and gradual decline in survival butstill maintained the highest survival throughout this exper-iment (Figure 2(a)) The 6∘C treatment had high mortality5 days after reaching 6∘C and 100 mortality on day 30(Figure 2(a)) End-point survival also revealed significantdifferences among temperature treatments (F

312= 398 119901 =

0035 Table 2) Overall M charruana had low survival incold temperatures with 38 as the highestmean final survivalat 13∘C

Warm experiments B and C investigated survival overtime at 20ndash36∘C and documented significant differencesamong treatments in both experiments (119901 lt 0001 Fig-ure 2(b) 119901 lt 00001 Figure 2(c)) The 20∘C (and 23∘C)treatment maintained the highest survival over 33 daysfollowed by the warmer temperature treatments (Figures 2(b)and 2(c)) Treatments at 28 31 and 36∘C experienced dra-matic mortality after reaching their respective experimentaltemperatures (Figures 2(b) and 2(c)) End-point survival inwarm experiments B and C showed that all treatments weresignificantly different from each other when compared withTukeyrsquos a posteriori tests (Exp B F

212= 25153 119901 lt 00001

Exp C F39

= 3518119901 lt 00001 Table 2)The 20∘C (and 23∘C)treatment had the highest final survivals at 83ndash88 followedby the higher temperatures ranging from 0 (at 36∘C) to 24(at 28∘C) (Figures 2(b) and 2(c))

32 Juvenile Perna viridis Salinity-Temperature InteractionExperiments Survivorship over time in the cold experimentwas significantly different overall for the tested salinity-temperature combinations (119901 lt 0001 Figure 3(a)) Thesurvivorship patterns showed low mortality during theadjustment period (15 days) with 77 or greater survival forall treatment combinations (Figure 3(a)) Two days after the

adjustment period ended all juveniles at 6∘C experienceda dramatic decline in survival (Figure 3(a)) At 9∘C noindividuals kept at salinities of 5 and 20 ppt survived past day28 but individuals held at a salinity of 20 ppt showed a moregradual decline At 20∘C survival was highest at salinities of36 and 20 ppt respectively and at salinity of 5 ppt mortalityincreased greatly from day 26

In the warm experiment with juvenile P viridis survivalanalysis showed a significant difference among the salinity-temperature treatments (119901 lt 0001 Figure 3(b)) Thesurvivorship patterns showed a gradual decline during the15-day adjustment period for all treatments except for asalinity of 20 ppt at 35∘C where survival declined morerapidly with only 50 alive on day 15 (Figure 3(b)) Slopessteadily declined to 100 mortality for low salinity and hightemperature combinations (Figure 3(b)) From both cold andwarm experiments for juvenile P viridis the survival patternsindicated that this species responded more quickly to theeffect of temperature than to salinity

When considering end-point survival for juvenile Pviridis there was no interaction between salinity and coldtemperature (F

418= 2132 119901 = 0119 Table 3) However

the data in the cold experiment showed a strong temperatureeffect (F

218= 7755 119901 = 0004) and an even stronger

salinity effect (F218

= 9940 119901 lt 0001) Individuals inthe cold experiment had the highest survival at the uppersalinity and temperature combinations and 100mortality inthe lower salinity and temperature combinations (Figure 4)Opposite to the cold experiment a significant interaction(F418

= 3366 119901 = 0032 Table 3) between salinity and warmtemperature was observed for final survival with juvenile Pviridis (Figure 4)

33 Adult Perna viridis Salinity-Temperature InteractionExperiments We found significant differences among treat-ments in the survivorship of adult P viridis in the coldexperiment (119901 lt 0001 Figure 3(c)) The combinationsthat included 6 and 9∘C media gradually declined until day15 followed by a more rapid decline to 6 survival or lessby day 28 (Figure 3(c)) The survivorship pattern for 20∘Ctreatments showed that low salinity had a negative effect onadult P viridis and that the difference in survivorship becamemore apparent during the last 8 days of the cold experiment(Figure 3(c))

For the warm experiment there was also a significantdifference among treatments in the survivorship of adult Pviridis (119901 lt 0001 Figure 3(d)) The warm temperaturesurvivorship pattern showed minimal decline during theadjustment period except at a salinity of 20 ppt at 31∘C(Figure 3(d)) Once the adjustment period ended at day 15more rapid declines in survival were observed at all testedtreatments when salinity was 5 ppt (Figure 3(d)) Adult Pviridis in both warm and cold experiments showed thattemperature affected survival more rapidly than changes insalinity

The final survival of adult P viridis in the cold experimentshowed that there was a significant interaction (F

418= 3480

119901 = 0028) between salinity and cold temperature (Table 3)The highest mortality was observed at the combination


Table 3 Two-way ANOVA tables interaction between salinity and temperature on nonnative mussels

Experiment Source Perna viridis Mytella charruanaDF MS 119865 119901 Significance DF MS 119865 119901 Significance

Juvenile Cold

Salinity 2 43370 9940 lt0001 lowast 2 1009 7977 0003 lowast

Temperature 2 55593 7755 0004 lowast 2 0127 63372 lt0001 lowast

Salinity times temperature 4 11926 2132 0119 ns 4 0041 2605 0071 nsError 18 5593 18 0016

Juvenile Warm

Salinity 2 47815 8208 0003 lowast 2 18 2978 0076 nsTemperature 2 30704 12782 lt0001 lowast 2 0103 52269 lt0001 lowast

Salinity times temperature 4 12593 3366 0032 lowast 4 0031 0914 0477 nsError 18 3741 18 0034

Adult Cold



Adult Warm

Salinity 2 0218 56194 lt0001 lowast 2 0471 10987 lt0001 lowast


Salinity times temperature 4 0088 3839 0020 lowast 4 0215 25217 lt0001 lowast

Error 18 0023 18 0009lowastSignificantly different at 119901 lt 005 level

of lowest salinities (5 20 ppt) and lowest temperatures (69∘C) (Figure 4) As for the warm experiment a significantinteraction (F

418= 3839 119901 = 0020) was also observed

in the final survival between salinity and warm temperature(Table 3) At 20∘C final survival was highest for all testedsalinities Unlike the survival of juvenile P viridis adults hadmoderate to high survival at warm temperatures and highsalinities (Figure 4)

34 Juvenile Mytella charruana Salinity-Temperature Interac-tion Experiments Survival analysis showed significant differ-ences among treatments in cold temperature and salinity onthe survivorship for juveniles (119901 lt 0001 Figure 5(a)) Inthis experiment the survivorship pattern showed a gradualdecline in survival for all treatment combinations through tothe end of the adjustment period (15 days) with 67or highersurvival (Figure 5(a)) All 6∘C treatments experienced rapiddeclines to 23 survival or lower during the postadjustmentperiod (Figure 5(a)) An intermediate gradual decline wasobserved at 9∘C with the highest survival in the lowestsalinity treatment (Figure 5(a)) At 20∘C the survival patternremained greater than 70 for all tested salinities (Fig-ure 5(a))

In the warm experiment survival analysis showed sig-nificant differences among treatments on survivorship forjuvenileM charruana (119901 lt 0001 Figure 5(b)) Similar grad-ual survivorship patterns were observed for all treatmentsbefore the adjustment period ended (Figure 5(b)) Treatmentsat 31 and 33∘C experienced a rapid decline in survival afteradjustment (Figure 5(b)) At 20∘C survival was 90or higheron day 28 The mortality of juvenile M charruana increasedas the temperature increased with the highest mortality inthe uppermost tested salinity

With final survival in the cold experiment there were nosignificant interactive effects between salinity and tempera-ture on juvenile M charruana (F

418= 2605 119901 = 0071

Table 3)There were however a significantmain effect of coldtemperature and a significant salinity effect on juveniles (F

218

= 63372 119901 lt 0001 F218

= 7977 119901 = 0003 resp Table 3)In the warm experiment there were also no significantinteractive effects between salinity and temperature on thefinal survival for juvenile M charruana (F

418= 0914 119901 =

0477) but a significant main effect of temperature (F218

=52269119901 lt 0001)Thefinal survival of juvenileM charruanashowed that they can endure temperatures that ranged from6 to 33∘C at a salinity of 5 ppt (Figure 4) The highest survivalwas at 20∘C and salinity of 225 ppt but mortality becamegreater as temperature and salinity decreased or increased(Figure 4) The highest mortality for juvenile M charruanawas observed at the combination of the highest salinity withlowest and highest tested temperatures

35 Adult Mytella charruana Salinity-Temperature Interac-tion Experiments Survival analysis showed significant differ-ences within treatments of cold temperature and salinity (119901 lt0001 Figure 5(c)) on the survivorship of adultM charruanaBefore the adjustment period ended mortality was onlyobserved at 6∘C at salinities of 5 and 40 ppt After adjustment6∘C treatments all showed declines to 7 survival or less byday 28 with survival at the highest salinity declining mostrapidly (Figure 5(c)) High survival (gt93) was observedat the end of the experiment at 20∘C at all tested salinities(Figure 5(c))

In the warm experiment with adult M charruana sur-vival analysis showed significant differences within treat-ments on survivorship (119901 lt 00001 Figure 5(d)) There


0

20

40

60

80

100

69203135

5

20

36

Mus

sel s

urvi

ved

()

Juvenile

Temperature (∘C)

Salin

ities

0

20

40

60

80

100

69203135

5

20

36

Mus

sels

surv

ived

()

AdultP viridis P viridis

Temperature (∘C)

Salin

ities

0

20

40

60

80

100

69203133

5

225

40

Mus

sels

surv

ived

()

Adult

Temperature (∘C)

Salin

ities

Salin

ities

0

20

40

60

80

100

69203133

5

225

40

Mus

sels

surv

ived

()

M charruana M charruana Juvenile

Temperature (∘C)

Figure 4 Final survival of Perna viridis in the salinity-temperature interaction experiments for juveniles and adults Final survival ofMytellacharruana in the salinity-temperature interaction experiments for juveniles and adults

was no mortality before the end of the adjustment period(Figure 5(d)) Individuals kept in warmer temperatures andsalinity of 40 showed rapid declines in survival beginning onday 17 Survival was 97 or higher in all treatments at 20∘C(Figure 5(d))

For the final cold survival experiment there was nosignificant interaction (F

418= 1653 119901 = 0205) between

salinity and temperature for adult M charruana though asignificant temperature effect (F

218= 148361 119901 lt 0001) was

observed (Table 3) In the warm temperature experiment asignificant interaction effect between salinity and tempera-ture (F

418= 25217119901 lt 0001) on the final survival of adultM

charruanawas observed (Table 3) Survival approached 100at 20∘C survival decreased both with reduced temperature

and with increased salinity (Figure 4) The highest mortalitywas observed at combinations of high salinity and hightemperature and of low salinity and low temperature

4 Discussion

Environmental factors influence every species differently andtheir impact can change throughout the life history of anorganism depending on body size and life stage [46 47]and is particularly relevant for nonnative species followingintroduction (eg [48ndash50]) It is important to evaluate thesynergistic effects of abiotic factors of all life stages to enableresearchers to predict range expansions and survival ofinvasive species [51ndash53] and to determine whether species


Juve

nile

sur

viva

l ove

r tim

e (

)

0102030405060708090

100Cold

Mytella charruana

(Days)0 4 8 12 16 20 24 28

BC

F

CD DEE

AB AB

ABA

5 6∘C225 6∘C40 6∘C5 9∘C225 9∘C

40 9∘C5 20∘C225 20∘C40 20∘C

(a)Ju

veni

le s

urvi

val o

ver t

ime (

)

0102030405060708090

100Warm

Mytella charruana

(Days)0 4 8 12 16 20 24 28

B

A

A DA

B C

BCD

9 20∘C225 20∘C40 20∘C9 31∘C225 31∘C

40 31∘C9 33∘C225 33∘C40 33∘C

(b)

ColdMytella charruana

(Days)0 4 8 12 16 20 24 28

Adul

t su

rviv

al o

ver t

ime (

)

0102030405060708090

100

B

D

CD CCD

C AB

AA

5 6∘C225 6∘C40 6∘C5 9∘C225 9∘C

40 9∘C5 20∘C225 20∘C40 20∘C

(c)

WarmMytella charruana

Adul

t su

rviv

al o

ver t

ime (

)

0102030405060708090

100

(Days)0 4 8 12 16 20 24 28

A

A

A BA

A B

AA

5 20∘C225 20∘C40 20∘C5 31∘C225 31∘C

40 31∘C5 33∘C225 33∘C40 33∘C

(d)

Figure 5 Survivorship curves over 28 days for Mytella charruana in the salinity-temperature interaction experiments (a) Juvenile coldexperiment (temperatures 6 9 and 20∘C salinities 5 225 and 40 ppt) (b) juvenile warm experiment (temperatures 20 31 and 33∘Csalinities 9 225 and 40 ppt) (c) adult cold experiment (temperatures 6 9 and 20∘C salinities 5 225 and 40 ppt) and (d) adult warmexperiment (temperatures 20 31 and 33∘C salinities 5 225 and 40 ppt) Upper case letters indicate significant differences of119901 le 005 amongtreatments using survival analysis Dotted lines show the end of the 15-day temperature adjustment periods

are evolving in their invaded range based on altered abioticconditions (eg [54]) In light of the recently characterizednegative impacts of Perna viridis and Mytella charruana onthe ecologically and economically important eastern oysterCrassostrea virginica [12] we compare our results to otherstudies and the implications of these results to the long-termpersistence and spread of these invasive species

Studies that considered body size of the invasive musselPerna viridis found that larger individuals could withstandabiotic stressors better than small-sized mussels [27 48]This pattern has also been shown for Perna viridis whenexposed to cold air temperatures that coincided with lowtides higher survival was documented for larger individuals[29] In contrast Yuan et al [24] found that small M


charruana survived a broader range of salinities than largerindividuals further indicating the importance of consideringbody size in understanding environmental gradients Ourcurrent study found that adult P viridis survived differentlyacross salinities and temperatures than juveniles (Figure 4)The most dramatic differences were that adult P viridis didnot survive or barely survived at the colder temperaturesregardless of salinity whereas juveniles were able to survive atcolder temperatures in the high salinity trials Furthermoresurvival was lowest for juvenile P viridis at the highest testedtemperature and lowest tested salinity (Figure 4) This is theopposite of Urian et al [29] who found large-sized P viridishad a greater survival than small-sized individuals whenexposed to cold air temperaturesThe difference between ourstudy and Urian et al [29] could reflect submersion (ourstudy) versus aerial exposure

Survival across salinities and temperatures appears to bespecies-specific for marine molluscs (eg [16 20 21]) Kinne[47] found several sessile and low-mobility invertebrates hadgreater survival success at low salinity and low temperaturesthan all other relative temperature and salinity combinationsLikewise Andrews et al [55] considered the oyster Cras-sostrea virginica to survive for long time periods in a stateof ldquonarcosisrdquo in low salinity and low temperature conditionsOther studies have shown the reverse where survival washigher at low salinityhigh temperature and high salinitylowtemperature combinations (eg [56 57]) For example astudy on the nonnative mollusc Mytilopsis leucophaeata inEurope found that embryonic mortality was high with thecombination of high salinity and low temperature as wellas with low salinity and high temperature [57] Salinityand temperature impacted the survival of P viridis and Mcharruana in different ways Perna viridis was influenced byboth salinity and temperature Perna viridis survived at awide range of temperatures from when the salinity was highyet as salinity decreased the range of temperatures at whichthe species survived became narrower In contrast we foundthat the survival of M charruana was primarily driven bytemperature Interestingly both species had some survivalat all tested salinities (P viridis 5ndash36 ppt M charruana 5ndash40 ppt) and at all tested temperatures (6ndash3335∘C Figure 4)Little is known about the salinity and temperature ranges ofMytella charruana in their native habitats but P viridis hadbeen described in waters where salinities ranged from 27 to33 ppt and where temperatures ranged from 26 to 32∘C in thePhilippines [25]

Temperature changes killedmore rapidly than salinity forboth species By analyzing survivorship curves rather thanjust final survival for a suite of temperatures and salinitiesfound within the southeastern United States we determinedthat temperature affected survival more quickly than salinityfor both species investigated hereThe temperature effect wasexpressed as a sharp drop in a survivorship curve shortly aftera salinity-temperature adjustment whereas salinity effectstended to result in slower declines in survivorship Manypublished studies that examine the importance of abioticvariables on marine invertebrates have focused on the finalresponse by the organism without considering the rateof mortality For example survival physiological response

reproduction and growth are often recorded but these resultsdo not incorporate the timing of the response to each abioticeffect [19 21 57ndash60] Schneider [61] however investigated theeffects of temperature and exposure to air on the survivorshipcurves for two intertidal species (Mytilus galloprovincialisMtrossulus) and found species-specific differences with rapidsurvival declines evident at the highest tested temperatures

Many nonnative species are more tolerant of environ-mental stressors than their native taxonomically relatedequivalents The success of nonnative species was demon-strated in Lenz et al [62] where the authors compared fivenative and nonnative marine invertebrate pairs on a globalscale covering five biogeographic regions and found that thenonnatives (bivalves P viridis Isognomon bicolor and Cras-sostrea gigas ascidian Didemnum vexillum and crustaceanGammarus tigrinus) had higher survival and wider toleranceranges than the paired native species (bivalves Brachidontesexustus Perna and Saccostrea glomerata ascidianDiplosomalisterianum and crustacean Gammarus tigrinus) Similarpatterns have been found in other taxa (eg mammals andbirds [63] plants [64]) Perna viridis andM charruana werefound along the southeastern USA Atlantic coast in shallowsubtidal regions on docks alongside the native bivalve mol-luscs Brachidontes exustus and Geukensia demissa as well aswith the native oysterCrassostrea virginicaOur study showedthat P viridis and M charruana did not have wider survivaltolerance ranges for salinity and temperature than those ofthese native species [65] while Brodsky et al [66] showedthat native G demissa continued to produce byssal threadsat lower temperatures than M charruana These resultssuggest that P viridis and M charruana are not adapted tolocal environmental extremes In the invaded region a fieldstudy on P viridis and M charruana reported that no liveindividuals were found during the winter of 20092010 atlocations where both were previously established [15] Thatwinter was one of the coldest on record and had an unusuallylong duration of freezing temperatures in the southeasternUnited States [67] Both species were subsequently reportedin the southeastern United States after this cold event [15]but whether these species reestablished via newly introducedpropagules or survived the cold weather event is unknown

In summary our study provides a greater understandingof how abiotic factors interactively affect the survival ofnonnative P viridis and M charruana Both species weretolerant of many salinity-temperature combinations foundin the southeastern United States and thus should be ableto both persist in their present invaded range and expandinto new estuaries and bays along the Atlantic coastline ofFlorida andwest within the Gulf ofMexico Freezes howevercould thwart expansions into colder waters Given that bothspecies negatively impact the eastern oyster C virginica [12]and possibly many other native species it is important tocontinue to study their physiological ecology and any changesin distributions over time

Competing Interests

The authors declare there is no conflict of interests regardingthe publication of this paper


Acknowledgments

Funding was provided by USDA NIFA Award no 2008-32320-04574 the Indian River Lagoon National EstuaryProgram The Nature Conservancy and the University ofCentral Florida Many thanks to M Achenie S BolivarJ Bridges M Donnelly S Garvis K Grablow V Lee JLedgard J Leissing J Manis E Nash R Odom E OlsonD Pasiechnik B Rodenbeck P Sacks L Sinnett S Smith JSolomon A Stenyakina L Tormoen M Tye AWay andWWinterhalter for assisting with collecting laboratory workand reading paper drafts

References

[1] G M Ruiz J T Carlton E D Grosholz and A H HinesldquoGlobal invasions of marine and estuarine habitats by non-indigenous species mechanisms extent and consequencesrdquoAmerican Zoologist vol 37 no 6 pp 621ndash632 1997

[2] M Enserink ldquoPredicting invasions biological invaders sweeprdquoScience vol 285 pp 1834ndash1836 1999

[3] G M Ruiz P W Fofonoff J T Carlton M J Wonhamand A H Hines ldquoInvasion of coastal marine communitiesin North America apparent patterns processes and biasesrdquoAnnual Review of Ecology and Systematics vol 31 pp 481ndash5312000

[4] I C Davidson C W Brown M D Sytsma and G M RuizldquoThe role of containerships as transfer mechanisms of marinebiofouling speciesrdquo Biofouling vol 25 no 7 pp 645ndash655 2009

[5] D K Padilla and S L Williams ldquoBeyond ballast water aquar-ium and ornamental trades as sources of invasive species inaquatic ecosystemsrdquo Frontiers in Ecology and the Environmentvol 2 no 3 pp 131ndash138 2004

[6] L J Walters K R Brown W T Stam and J L Olsen ldquoE-commerce and Caulerpa unregulated dispersal of invasivespeciesrdquo Frontiers in Ecology and the Environment vol 4 no2 pp 75ndash79 2006

[7] C C Murray E A Pakhomov and T W Therriault ldquoRecre-ational boating a large unregulated vector transporting marineinvasive speciesrdquo Diversity and Distributions vol 17 no 6 pp1161ndash1172 2011

[8] V Crego-Prieto A Ardura F Juanes A Roca J S Taylor andE Garcia-Vazquez ldquoAquaculture and the spread of introducedmussel genes in British Columbiardquo Biological Invasions vol 17no 7 pp 2011ndash2026 2015

[9] A RicCIardi ldquoGlobal range expansion of the Asian musselLimnoperna fortunei (Mytilidae) another fouling threat tofreshwater systemsrdquo Biofouling vol 13 no 2 pp 97ndash106 1998

[10] N Bax AWilliamsonM Aguero E Gonzalez andW GeevesldquoMarine invasive alien species a threat to global biodiversityrdquoMarine Policy vol 27 no 4 pp 313ndash323 2003

[11] G M Branch and C N Steffani ldquoCan we predict the effectsof alien species A case-history of the invasion of South AfricabyMytilus galloprovincialis (Lamarck)rdquo Journal of ExperimentalMarine Biology and Ecology vol 300 no 1-2 pp 189ndash215 2004

[12] W S Yuan E A Hoffman and L J Walters ldquoEffects ofnonnative invertebrates on two life stages of the native easternoyster Crassostrea virginicardquo Biological Invasions vol 18 no 3pp 689ndash701 2016

[13] D Pimentel R Zuniga and D Morrison ldquoUpdate on the envi-ronmental and economic costs associated with alien-invasive

species in the United Statesrdquo Ecological Economics vol 52 no3 pp 273ndash288 2005

[14] M L Boudreaux and L JWalters ldquoMytella charruana (BivalviaMytilidae) a new invasive bivalve in Mosquito LagoonFloridardquo Nautilus vol 120 no 1 pp 34ndash36 2006

[15] S S Spinuzzi K R Schneider L J Walters W S Yuanand E A Hoffman ldquoTracking the distribution of non-nativemarine invertebrates (Mytella charruana Perna viridis andMegabalanus coccopoma) along the south-easternUSArdquoMarineBiodiversity Records vol 6 article e55 13 pages 2013

[16] S E Shumway ldquoNatural environmental factorsrdquo inThe EasternOyster Crassostrea virginica V S Kennedy R I E Newelland A F Eble Eds pp 467ndash514 Maryland Sea Grant CollegeCollege Park Md USA 1996

[17] S T Tettelbach and E W Rhodes ldquoCombined effects oftemperature and salinity on embryos and larvae of the northernbay scallop Argopecten irradiansrdquoMarine Biology vol 63 no 3pp 249ndash256 1981

[18] G S Rupp andG J Parsons ldquoEffects of salinity and temperatureon the survival and byssal attachment of the lionrsquos paw scallopNodipecten nodosus at its southern distribution limitrdquo Journalof Experimental Marine Biology and Ecology vol 309 no 2 pp173ndash198 2004

[19] R Przeslawski A R Davis and K Benkendorff ldquoSynergisticeffects associated with climate change and the development ofrocky shore molluscsrdquo Global Change Biology vol 11 no 3 pp515ndash522 2005

[20] E His R Robert and A Dinet ldquoCombined effects of tempera-ture and salinity on fed and starved larvae of the mediterraneanmussel Mytilus galloprovincialis and the Japanese oyster Cras-sostrea gigasrdquoMarine Biology vol 100 no 4 pp 455ndash463 1989

[21] G Sara C Romano JWiddows and F J Staff ldquoEffect of salinityand temperature on feeding physiology and scope for growthof an invasive species (Brachidontes pharaonismdashMOLLUSCABIVALVIA) within the Mediterranean seardquo Journal of Experi-mentalMarine Biology andEcology vol 363 no 1-2 pp 130ndash1362008

[22] M I Segnini de Bravo K S Chung and J E Perez ldquoSalinityand temperature tolerances of the green and brown musselsPerna viridis and Perna Perna (Bivalvia Mytilidae)rdquo Revista deBiologia Tropical vol 46 supplement 5 pp 121ndash125 1998

[23] R Manoj Nair and K K Appukuttan ldquoEffect of temperatureon the development growth survival and settlement of greenmussel Perna viridis (Linnaeus 1758)rdquo Aquaculture Researchvol 34 no 12 pp 1037ndash1045 2003

[24] W Yuan L J Walters K R Schneider and E A HoffmanldquoExploring the survival threshold a study of salinity toleranceof the nonnative musselMytella charruanardquo Journal of ShellfishResearch vol 29 no 2 pp 415ndash422 2010

[25] J M Vakily The Biology and Culture of Mussels of the GenusPerna International Center for Living Aquatic Resources Man-agement Manila Philippines 1989

[26] P M Sivalingam ldquoAquaculture of the green mussel Mytilusviridis Linnaeus in Malaysiardquo Aquaculture vol 11 no 4 pp297ndash312 1977

[27] K S Chung and A Acuna ldquoUpper temperature tolerance limitof mussel Pernardquo Bulletin of the Japanese Society of ScientificFisheries vol 47 no 3 p 441 1981

[28] K V K Nair and P Murugan ldquoBiofouling by Perna viridis ina deep submarine tunnel system at Lalpakkamrdquo Journal of theMarine Biological Association of India vol 33 pp 366ndash377 1991


[29] AGUrian J DHatle andM RGilg ldquoThermal constraints forrange expansion of the invasive green mussel Perna viridis inthe southeasternUnited Statesrdquo Journal of Experimental ZoologyPart A Ecological Genetics and Physiology vol 315 no 1 pp 12ndash21 2011

[30] S E Siddall ldquoA classification of the genus Pernardquo Bulletin ofMarine Science vol 30 no 4 pp 858ndash870 1980

[31] D A Ingrao P M Mikkelsen and DW Hicks ldquoAnother intro-duced marine mollusk in the Gulf of Mexico the Indo-Pacificgreen mussel Perna viridis in Tampa bay Floridardquo Journal ofShellfish Research vol 20 no 1 pp 13ndash19 2001

[32] P Baker J S Fajans W S Arnold D A Ingrao D C Marelliand S M Baker ldquoRange and dispersal of a tropical marineinvader the Asian green mussel Perna viridis in subtropicalwaters of the southeastern United Statesrdquo Journal of ShellfishResearch vol 26 no 2 pp 345ndash355 2007

[33] IFAS Green Mussels 2007 httpsfrcufledugreenmussel[34] P Baker J S Fajans and S M Baker ldquoHabitat dominance

of a nonindigenous tropical bivalve Perna viridis (Linnaeus1758) in a subtropical estuary in the Gulf of Mexicordquo Journalof Molluscan Studies vol 78 no 1 pp 28ndash33 2012

[35] A M Keen Sea Shells of Tropical West America MarineMollusks from Baja California to Peru Stanford UniversityPress Stanford Calif USA 1971

[36] J T Carlton ldquoIntroduced marine and estuarine mollusksof North America an end-of-the-20th- century perspectiverdquoJournal of Shellfish Research vol 11 pp 489ndash505 1992

[37] P Szefer J Gełdon A A Ali F Paez Osuna A C Ruiz-Fernandes and S R G Guerrero Galvan ldquoDistribution andassociation of trace metals in soft tissue and byssus of Mytellastrigata and other benthal organisms from Mazatlan HarbourMangrove Lagoon of the northwest coast of Mexicordquo Environ-ment International vol 24 no 3 pp 359ndash374 1998

[38] G Boehs T Absher and A da Cruz-Kaled ldquoComposition anddistribution of benthic mollusks on intertidal flats of ParanaguaBay (Parana Brazil)rdquo Scientifica Marina vol 68 pp 537ndash5432004

[39] H G Lee Immigrant Mussel Settles in Northside Generator TheShell-O-Gram vol 28 Jacksonville Shell Club Jacksonville FlaUSA 1987

[40] O M Pereira R C Hilberath P R A C Ansarah and MS N Galvao ldquoProduction estimate of Mytella falcata and Mguyanensis in natural beds of Ilha Comprida Estuary (Sao PauloBrazil)rdquo Boletin Do Instituto De Pesca vol 29 pp 139ndash149 2003

[41] A Stenyakina L J Walters E A Hoffman and C CalestanildquoFood availability and sex reversal in Mytella charruana anintroduced bivalve in the southeasternUnited StatesrdquoMolecularReproduction and Development vol 77 no 3 pp 222ndash230 2010

[42] NOAA National Centers for environmental information 2016httpwwwncdcnoaagov

[43] A J Power R L Walker K Payne and D Hurley ldquoFirstoccurrence of the nonindigenous green mussel Perna viridis(Linnaeus 1758) in coastal Georgia United Statesrdquo Journal ofShellfish Research vol 23 no 3 pp 741ndash744 2004

[44] JMP Version 10 SAS Institute Cary NC USA 1989ndash2012[45] E L Kaplan and P Meier ldquoNonparametric estimation from

incomplete observationsrdquo Journal of the American StatisticalAssociation vol 53 no 282 pp 457ndash481 1958

[46] L W Hutchins ldquoThe bases for temperature zonation in geo-graphical distributionrdquo Ecological Monographs vol 17 no 3 pp325ndash335 1947

[47] O Kinne ldquoTemperature-invertebratesrdquo in Marine Ecology IPart 1 O Kinne Ed pp 405ndash514 Wiley-Interscience LondonUK 1970

[48] D W Hicks and R F McMahon ldquoRespiratory responses totemperature and hypoxia in the nonindigenous brown musselPerna perna (Bivalvia Mytilidae) from the Gulf of MexicordquoJournal of Experimental Marine Biology and Ecology vol 277no 1 pp 61ndash78 2002

[49] A Nasrolahi C Pansch M Lenz and M Wahl ldquoBeing youngin a changing world how temperature and salinity changesinteractively modify the performance of larval stages of thebarnacle Amphibalanus improvisusrdquo Marine Biology vol 159no 2 pp 331ndash340 2012

[50] M C Pineda C D McQuaid X Turon S Lopez-Legentil VOrdonez and M Rius ldquoTough adults frail babies an analysisof stress sensitivity across early life-history stages of widelyintroduced marine invertebratesrdquo PLoS ONE vol 7 no 10Article ID e46672 2012

[51] C E Braby and G N Somero ldquoFollowing the heart temper-ature and salinity effects on heart rate in native and invasivespecies of bluemussels (genusMytilus)rdquo Journal of ExperimentalBiology vol 209 no 13 pp 2554ndash2566 2006

[52] E J Hawes andD L Parrish ldquoUsing abiotic and biotic factors topredict the range expansion of white perch in Lake ChamplainrdquoJournal of Great Lakes Research vol 29 no 2 pp 268ndash279 2003

[53] S B Menke and D A Holway ldquoAbiotic factors control invasionby Argentine ants at the community scalerdquo Journal of AnimalEcology vol 75 no 2 pp 368ndash376 2006

[54] K A Medley ldquoNiche shifts during the global invasion ofthe Asian tiger mosquito Aedes albopictus Skuse (Culicidae)revealed by reciprocal distribution modelsrdquo Global Ecology andBiogeography vol 19 no 1 pp 122ndash133 2010

[55] J Andrews D Haven and D Quayle ldquoFreshwater kill ofoysters (Crassostrea virginica) in James River Virginia 1958rdquoProceedings of the National Shellfish Association vol 49 pp 29ndash49 1959

[56] M H Brenko and A Calabrese ldquoThe combined effects ofsalinity and temperature on larvae of the musselMytilus edulisrdquoMarine Biology vol 4 no 3 pp 224ndash226 1969

[57] A Verween M Vincx and S Degraer ldquoThe effect of tempera-ture and salinity on the survival ofMytilopsis leucophaeata lar-vae (Mollusca Bivalvia) the search for environmental limitsrdquoJournal of Experimental Marine Biology and Ecology vol 348no 1-2 pp 111ndash120 2007

[58] Y J Xu and J Lin ldquoEffect of temperature salinity and lightintensity on the growth of the green macroalga Chaetomorphalinumrdquo Journal of the World Aquaculture Society vol 39 no 6pp 847ndash851 2008

[59] A Epelbaum L M Herborg T W Therriault and C MPearce ldquoTemperature and salinity effects on growth survivalreproduction and potential distribution of two non-indigenousbotryllid ascidians in British Columbiardquo Journal of Experimen-tal Marine Biology and Ecology vol 369 no 1 pp 43ndash52 2009

[60] B L Lockwood and G N Somero ldquoInvasive and native bluemussels (genus Mytilus) on the California coast the role ofphysiology in a biological invasionrdquo Journal of ExperimentalMarine Biology and Ecology vol 400 no 1-2 pp 167ndash174 2011

[61] K R Schneider ldquoHeat stress in the intertidal comparingsurvival and growth of an invasive and native mussel under avariety of thermal conditionsrdquo Biological Bulletin vol 215 no3 pp 253ndash264 2008


[62] M Lenz B A P da Gama N V Gerner et al ldquoNon-nativemarine invertebrates are more tolerant towards environmentalstress than taxonomically related native species results from aglobally replicated studyrdquo Environmental Research vol 111 no7 pp 943ndash952 2011

[63] J M Jeschke and D L Strayer ldquoDeterminants of vertebrateinvasion success in Europe and North AmericardquoGlobal ChangeBiology vol 12 no 9 pp 1608ndash1619 2006

[64] M van Kleunen E Weber and M Fischer ldquoA meta-analysisof trait differences between invasive and non-invasive plantspeciesrdquo Ecology Letters vol 13 no 2 pp 235ndash245 2010

[65] Smithsonian Marine Station at Fort Pierce Indian RiverLagoon Species Inventory 2011 httpwwwsmssieduirlspecindexhtm

[66] S Brodsky L Walters and E Hoffman ldquoCold temperatureeffects on byssal thread production by the native musselGeukensia demissa versus the non-native mussel Mytella char-ruanardquo University of Central Florida Undergraduate ResearchJournal vol 5 no 1 pp 1ndash10 2011

[67] NOAA National Weather Service Weather Forecast OfficeJacksonville FL 2010 A year of historical climate extremesin northeast Florida and southeast Georgia httpwwwsrhnoaagovjaxn=2010 summary

Submit your manuscripts athttpwwwhindawicom

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

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International Journal of

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Zoology


Molecular Biology International

GenomicsInternational Journal of


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BioinformaticsAdvances in

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Evolutionary BiologyInternational Journal of



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ArchaeaHindawi Publishing Corporationhttpwwwhindawicom Volume 2014


Genetics Research International


Advances in

Virolog y

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Nucleic AcidsJournal of

Volume 2014

Stem CellsInternational



Enzyme Research



Microbiology


testing a range of salinities and temperatures to determine itsphysiological tolerance for feeding These researchers deter-mined that the clearance rate of B pharaonis declined underconditions of low salinity and low temperature Likewise Hiset al [20] examined the combined effects of temperature andsalinity on the invasive Japanese oyster C gigas in France anddetermined that the oyster exhibited the best growth at highertemperatures

Previous studies have independently investigated the roleof salinity and temperature in survival of P viridis and therole of salinity in survival in M charruana [22ndash24] Pernaviridis has been documented in salinities ranging from 19 to44 ppt in its native habitat [25] Segnini de Bravo et al [22]determined that P viridis could tolerate an even wider rangeof salinities under laboratory conditions (25∘C) and the lowerand upper lethal salinities were 0 and 64 ppt respectivelyPerna viridis survived at temperatures ranging from 10 to42∘C in the laboratory [25ndash29]This broad temperature rangeshould enable P viridis to survive in both tropical andtemperate zones

Mytella charruana survived in salinities ranging from2 to 40 ppt at 20∘C [24] Not much is known about thethermal tolerance for M charruana so the first goal of ourstudy was to address how temperature influenced survivalin M charruana independent of salinity By addressing thisknowledge gap we were then able to determine appropriatetemperature ranges for our salinity-temperature interactionexperiments

The overall objective of this study was to investigate thesimultaneous effects of salinity and temperature on nonnativePerna viridis and Mytella charruana to better understandtheir respective invasion potentials Specifically we addressedthe following questions (1)What is the temperature tolerancerange for survival of Mytella charruana (2) How do salinityand temperature interact to influence the survival of adultand juvenile Perna viridis (3) How does the interactionof salinity and temperature influence the survival of adultand juvenile Mytella charruana These results will aid inpredicting suitable environments where adult and juvenileindividuals of both molluscs may become established andpredict future range expansions

2 Materials and Methods

21 Study Species Perna viridis the Asian green mussel isnative to the Indo-Pacific region along the Southeast Asiancoastline from the Arabian Gulf to southern China andsouthern Japan [30] and was first documented in the UnitedStates in 1999 in the cooling system of an electrical powerplant in Tampa Florida [31] Currently P viridis is wellestablished in the United States and extends from the TenThousand Islands to the northern Gulf of Mexico on the westcoast of Florida and along the eastern United States coastlinefrom Stuart FL (27∘ 098311015840N 80∘ 116041015840W) to northernGeorgia (32∘ 012061015840N 80∘ 595691015840W) [15 32 33] In TampaBay P viridis had the highest density of all observed locationswithin the United States with 2505 individuals mminus2 [34]

Mytella charruana commonly known as the charru mus-sel or sururu is native to the Galapagos Islands the Pacific

coast from Mexico to Ecuador [35ndash38] and the Atlanticcoast from Argentina to Venezuela [39] Like P viridis thediscovery ofM charruana in the United States occurred at apower plantMytella charruana was reported in JacksonvilleFL in 1987 it was considered to be locally extirpated by spring1988 [39] In 2004 however M charruana was found inMosquito Lagoon 212 km south of Jacksonville [14] Mytellacharruana has now been recorded along the southeasternUnited States coast from Titusville FL (28∘ 372211015840N 80∘483711015840W) to Charleston SC (32∘ 464521015840N 79∘ 575391015840W)[15] Densities of this nonnative species in the United Stateshave been measured up to sim12 individuals mminus2 [15] Thisdensity is much lower than that in their native habitats whereM charruana densities as high as 11036 mussels mminus2 havebeen reported [40]

22 Thermal Tolerance of Mytella charruana Mytella char-ruana (20ndash54mm) were collected in cold and warm monthsbetweenDecember 2007 andOctober 2009 from two floatingdock sites (Arlington Lions Club and Lonnie Wurn) inJacksonville FL (Table 1(a) Figure 1)The location of samplescollected for our experiments varied because we could notcollect the 300ndash400 mussels needed per experiment from asingle site Stenyakina et al [41] found thatM charruanaweresexually mature at 125mm For thermal tolerance trials weused adult mussels that were ge20mm

Individuals were acclimated to the laboratory environ-ment at 20∘C for 13 days in a recirculating aquaria (151 L)with water from the collection site This temperature waschosen as an intermediate for acclimation of all individualsas we previously determined that this species placed inwater at this temperature always survived in high numbersregardless of whether it represented a temperature increase ordecrease from field conditions In Florida between 1981 and2010 the temperature ranged from 3 to 26∘C during January(coldest month) and from 19 to 35∘C in July (warmest month)[42] Thirteen days was used as the acclimation period asit represented sufficient time for any mortality associatedwith collection to occur Prior to running these trials wedocumented that individuals that survived at least 13 dayswould survive for months in 20∘C waters (Yuan pers obs)Additional details of each trial are listed in Table 1(a)

After acclimation groups of 25 adult individuals rangingin length from 20 to 54mm were placed in separate aeratedaquaria (21 L) that contained 14 L of water in a ratio of 25collection site water 75 artificial seawater (Instant Oceansalts and deionized water) that maintained the salinity fromthe collection site It was not practical to collect sufficientnatural seawater for all trials Survival in preliminary trialswas 100 with this ratio of seawater to natural seawater(Yuan pers obs) We divided the range of experimentaltemperatures into two seasons cold for winter and warm forsummer conditions To avoid shocking the M charruanacold and warm experiments were correlated with field col-lection temperatures

Cold experiment A was conducted in a walk-in freezer at6∘C at the University of Central Florida at 6 9 11 and 13∘CWarm experiment B examined survival at 20 31 and 36∘Cwhile experiment C investigated survival over a narrower


Table1(a)S

ummaryo


ents

(b)S

ummaryo

fPerna

virid

isand(c)M


interactionexperim

ents

Acclimationperio

dwasthen

umbero

fdaysinho

ldingtanks

priortos

tartofexperim

ents

Adjustm

entperiodwasthen

umbero

fdaystotransition

fromho

ldingtanktem

perature(20∘C)

totre

atmenttem

peratures

Experim

entp

eriodwas

then

umbero

fdaystria

lran

inclu

ding

adjustmentp

eriodCa


ndto

thee

xperim


Figures2

3and

5

(a)Mytellacharruana

Experim

ents

Collectionsites

Latitud

eamplong

itude

Collection

temperatures

(∘ C)

Mussel

size(mm)

Collection

salin

ity(ppt)

Acclimation

perio

d(days)

Adjustm

ent

perio

d(days)

Experim

ent

perio

d(days)

Musselsper

tank

replicate

tank

s

Temperatures

teste

d(∘ C

)

Salin

ities

teste

d(ppt)

Adult

Arling

tonLion

sClub

3037

82721015840

N13

20ndash4

610

1315

33254

6911and

13NA

Acold

JacksonvilleFL

816208281015840W

Adult

Lonn

ieWurn

3037

57981015840

N28

22ndash54

813

1533

253

2031and36

NA

Bwarm

JacksonvilleFL

815851651015840W

Adult

Lonn

ieWurn

3037

57981015840

N27

25ndash4

610

1315

33254

202328and

31NA

Cwarm

JacksonvilleFL

8152

51651015840

W

(b)Pernavirid

is

Experim

ents

Collectionsites

Latitud

eamplong

itude

Collection

temperatures

(∘ C)

Mussel

size(mm)

Collection

salin

ity(ppt)

Acclimation

perio

d(days)

Adjustm

ent

perio

d(days)

Experim

ent

perio

d(days)

Musselsper

tank

replicate

tank

s

Temperatures

teste

d(∘ C

)

Salin

ities

teste

d(ppt)

Juvenile

Hugueno

tPark

304052371015840N

1012ndash38

365

1528

103

69and20

52036

Acold

JacksonvilleFL

81403211015840

WJuvenile

Hugueno

tPark

304052371015840N

1012ndash38

365

1528

103

2031and35

52036

Bwarm

JacksonvilleFL

81403211015840

WAd

ult

Mun

icipalMarina

298919731015840N

1858ndash114

355

1528

103

69and20

52035

Ccold

StA

ugustin

eFL

8131

00511015840

WAd

ult

Mun

icipalMarina

298919731015840N

2857ndash113

375

1528

103

2031and35

52037

Dw

arm

StA

ugustin

eFL

8131

00511015840

W

(c)Mytellacharruana

Experim

ents

Collectionsites

Latitud

eamplong

itude

Collection

temperatures

(∘ C)

Mussel

size(mm)

Collection

salin

ity(ppt)

Acclimation

perio

d(days)

Adjustm

ent

perio

d(days)

Experim

ent

perio

d(days)

Musselsper

tank

replicate

tank

s

Temperatures

teste

d(∘ C

)

Salin

ities

teste

d(ppt)

Juvenile

Fire

Station38

3039

00771015840

N17

5ndash12

57

1528

103

69and20

522540

Ecold

JacksonvilleFL

816415761015840W

Juvenile

Fire

Station38

3039

00771015840

N29

4ndash12

97

1528

103

2031and33

522540

Fwarm

JacksonvilleFL

816415761015840W

Adult

Fire

Station38

3039

00771015840

N16

20ndash38

57

1528

103

69and20

522540

Gcold

JacksonvilleFL

816415761015840W

Adult

Arling

tonLion

sClub

3037

82721015840

N28

20ndash31

57

1528

103

2031and33

522540

Hw

arm

JacksonvilleFL

816208281015840W


0 45 9 18

(km)

N

W E

S

0 105 210 420

(km)


(b)

(a)








0102030405060708090

100Su

rviv

al o

ver t

ime (

)

(Days)0 3 6 9 12 15 18 21 24 27 30 33

CB

BA

6∘C

9∘C

11∘C

13∘C

(a)

Surv

ival

ove

r tim

e (

)

0102030405060708090

100

(Days)0 3 6 9 12 15 18 21 24 27 30 33

AB

C20∘C

31∘C

36∘C

(b)

Surv

ival

ove

r tim

e (

)

(Days)0 3 6 9 12 15 18 21 24 27 30 33

0102030405060708090

100

AA

BC

20∘C

23∘C

28∘C

31∘C

(c)







Juve

nile

sur

viva

l ove

r tim

e (

)

0102030405060708090

100

(Days)0 4 8 12 16 20 24 28

ColdPerna viridis

DDCDCD

ABDABA

5 6∘C20 6∘C36 6∘C5 9∘C20 9∘C

36 9∘C5 20∘C20 20∘C36 20∘C

(a)Ju

veni

le s

urvi

val o

ver t

ime (

)

0102030405060708090

100

(Days)0 4 8 12 16 20 24 28

WarmPerna viridis

DACABEFBC

CDFCDEC

5 20∘C20 20∘C36 20∘C5 31∘C20 31∘C

36 31∘C5 35∘C20 35∘C36 35∘C

(b)

(Days)0 4 8 12 16 20 24 28

Adul

t su

rviv

al o

ver t

ime (

)

0102030405060708090

100Cold

Perna viridis

C

D

CD CDCD

CD A

BAB

5 6∘C20 6∘C35 6∘C5 9∘C20 9∘C

35 9∘C5 20∘C20 20∘C35 20∘C

(c)

Adul

t su

rviv

al o

ver t

ime (

)

0102030405060708090

100

(Days)0 4 8 12 16 20 24 28

WarmPerna viridis

CDBC

AA

B

DE

EF

C

5 20∘C20 20∘C37 20∘C5 31∘C20 31∘C

37 31∘C5 35∘C20 35∘C37 35∘C

(d)












3 Results


312= 398 119901 =



212= 25153 119901 lt 00001

Exp C F39






418= 2132 119901 = 0119 Table 3) However









418= 3480





Juvenile Cold




Juvenile Warm



Adult Cold



Adult Warm











418= 2605 119901 = 0071


218

= 63372 119901 lt 0001 F218


418= 0914 119901 =






0

20

40

60

80

100

69203135

5

20

36

Mus

sel s

urvi

ved

()

Juvenile

Temperature (∘C)

Salin

ities

0

20

40

60

80

100

69203135

5

20

36

Mus

sels

surv

ived

()


Temperature (∘C)

Salin

ities

0

20

40

60

80

100

69203133

5

225

40

Mus

sels

surv

ived

()

Adult

Temperature (∘C)

Salin

ities

Salin

ities

0

20

40

60

80

100

69203133

5

225

40

Mus

sels

surv

ived

()


Temperature (∘C)




418= 1653 119901 = 0205) between


218= 148361 119901 lt 0001) was





4 Discussion



Juve

nile

sur

viva

l ove

r tim

e (

)

0102030405060708090

100Cold

Mytella charruana

(Days)0 4 8 12 16 20 24 28

BC

F

CD DEE

AB AB

ABA

5 6∘C225 6∘C40 6∘C5 9∘C225 9∘C

40 9∘C5 20∘C225 20∘C40 20∘C

(a)Ju

veni

le s

urvi

val o

ver t

ime (

)

0102030405060708090

100Warm

Mytella charruana

(Days)0 4 8 12 16 20 24 28

B

A

A DA

B C

BCD

9 20∘C225 20∘C40 20∘C9 31∘C225 31∘C

40 31∘C9 33∘C225 33∘C40 33∘C

(b)


(Days)0 4 8 12 16 20 24 28

Adul

t su

rviv

al o

ver t

ime (

)

0102030405060708090

100

B

D

CD CCD

C AB

AA

5 6∘C225 6∘C40 6∘C5 9∘C225 9∘C

40 9∘C5 20∘C225 20∘C40 20∘C

(c)


Adul

t su

rviv

al o

ver t

ime (

)

0102030405060708090

100

(Days)0 4 8 12 16 20 24 28

A

A

A BA

A B

AA

5 20∘C225 20∘C40 20∘C5 31∘C225 31∘C

40 31∘C5 33∘C225 33∘C40 33∘C

(d)











Competing Interests



Acknowledgments


References














































































Volume 2014

Zoology






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology


Table1(a)S

ummaryo


ents

(b)S

ummaryo

fPerna

virid

isand(c)M


interactionexperim

ents

Acclimationperio

dwasthen

umbero

fdaysinho

ldingtanks

priortos

tartofexperim

ents

Adjustm

entperiodwasthen

umbero

fdaystotransition

fromho

ldingtanktem

perature(20∘C)

totre

atmenttem

peratures

Experim

entp

eriodwas

then

umbero

fdaystria

lran

inclu

ding

adjustmentp

eriodCa


ndto

thee

xperim


Figures2

3and

5

(a)Mytellacharruana

Experim

ents

Collectionsites

Latitud

eamplong

itude

Collection

temperatures

(∘ C)

Mussel

size(mm)

Collection

salin

ity(ppt)

Acclimation

perio

d(days)

Adjustm

ent

perio

d(days)

Experim

ent

perio

d(days)

Musselsper

tank

replicate

tank

s

Temperatures

teste

d(∘ C

)

Salin

ities

teste

d(ppt)

Adult

Arling

tonLion

sClub

3037

82721015840

N13

20ndash4

610

1315

33254

6911and

13NA

Acold

JacksonvilleFL

816208281015840W

Adult

Lonn

ieWurn

3037

57981015840

N28

22ndash54

813

1533

253

2031and36

NA

Bwarm

JacksonvilleFL

815851651015840W

Adult

Lonn

ieWurn

3037

57981015840

N27

25ndash4

610

1315

33254

202328and

31NA

Cwarm

JacksonvilleFL

8152

51651015840

W

(b)Pernavirid

is

Experim

ents

Collectionsites

Latitud

eamplong

itude

Collection

temperatures

(∘ C)

Mussel

size(mm)

Collection

salin

ity(ppt)

Acclimation

perio

d(days)

Adjustm

ent

perio

d(days)

Experim

ent

perio

d(days)

Musselsper

tank

replicate

tank

s

Temperatures

teste

d(∘ C

)

Salin

ities

teste

d(ppt)

Juvenile

Hugueno

tPark

304052371015840N

1012ndash38

365

1528

103

69and20

52036

Acold

JacksonvilleFL

81403211015840

WJuvenile

Hugueno

tPark

304052371015840N

1012ndash38

365

1528

103

2031and35

52036

Bwarm

JacksonvilleFL

81403211015840

WAd

ult

Mun

icipalMarina

298919731015840N

1858ndash114

355

1528

103

69and20

52035

Ccold

StA

ugustin

eFL

8131

00511015840

WAd

ult

Mun

icipalMarina

298919731015840N

2857ndash113

375

1528

103

2031and35

52037

Dw

arm

StA

ugustin

eFL

8131

00511015840

W

(c)Mytellacharruana

Experim

ents

Collectionsites

Latitud

eamplong

itude

Collection

temperatures

(∘ C)

Mussel

size(mm)

Collection

salin

ity(ppt)

Acclimation

perio

d(days)

Adjustm

ent

perio

d(days)

Experim

ent

perio

d(days)

Musselsper

tank

replicate

tank

s

Temperatures

teste

d(∘ C

)

Salin

ities

teste

d(ppt)

Juvenile

Fire

Station38

3039

00771015840

N17

5ndash12

57

1528

103

69and20

522540

Ecold

JacksonvilleFL

816415761015840W

Juvenile

Fire

Station38

3039

00771015840

N29

4ndash12

97

1528

103

2031and33

522540

Fwarm

JacksonvilleFL

816415761015840W

Adult

Fire

Station38

3039

00771015840

N16

20ndash38

57

1528

103

69and20

522540

Gcold

JacksonvilleFL

816415761015840W

Adult

Arling

tonLion

sClub

3037

82721015840

N28

20ndash31

57

1528

103

2031and33

522540

Hw

arm

JacksonvilleFL

816208281015840W


0 45 9 18

(km)

N

W E

S

0 105 210 420

(km)


(b)

(a)








0102030405060708090

100Su

rviv

al o

ver t

ime (

)

(Days)0 3 6 9 12 15 18 21 24 27 30 33

CB

BA

6∘C

9∘C

11∘C

13∘C

(a)

Surv

ival

ove

r tim

e (

)

0102030405060708090

100

(Days)0 3 6 9 12 15 18 21 24 27 30 33

AB

C20∘C

31∘C

36∘C

(b)

Surv

ival

ove

r tim

e (

)

(Days)0 3 6 9 12 15 18 21 24 27 30 33

0102030405060708090

100

AA

BC

20∘C

23∘C

28∘C

31∘C

(c)







Juve

nile

sur

viva

l ove

r tim

e (

)

0102030405060708090

100

(Days)0 4 8 12 16 20 24 28

ColdPerna viridis

DDCDCD

ABDABA

5 6∘C20 6∘C36 6∘C5 9∘C20 9∘C

36 9∘C5 20∘C20 20∘C36 20∘C

(a)Ju

veni

le s

urvi

val o

ver t

ime (

)

0102030405060708090

100

(Days)0 4 8 12 16 20 24 28

WarmPerna viridis

DACABEFBC

CDFCDEC

5 20∘C20 20∘C36 20∘C5 31∘C20 31∘C

36 31∘C5 35∘C20 35∘C36 35∘C

(b)

(Days)0 4 8 12 16 20 24 28

Adul

t su

rviv

al o

ver t

ime (

)

0102030405060708090

100Cold

Perna viridis

C

D

CD CDCD

CD A

BAB

5 6∘C20 6∘C35 6∘C5 9∘C20 9∘C

35 9∘C5 20∘C20 20∘C35 20∘C

(c)

Adul

t su

rviv

al o

ver t

ime (

)

0102030405060708090

100

(Days)0 4 8 12 16 20 24 28

WarmPerna viridis

CDBC

AA

B

DE

EF

C

5 20∘C20 20∘C37 20∘C5 31∘C20 31∘C

37 31∘C5 35∘C20 35∘C37 35∘C

(d)












3 Results


312= 398 119901 =



212= 25153 119901 lt 00001

Exp C F39






418= 2132 119901 = 0119 Table 3) However









418= 3480





Juvenile Cold




Juvenile Warm



Adult Cold



Adult Warm











418= 2605 119901 = 0071


218

= 63372 119901 lt 0001 F218


418= 0914 119901 =






0

20

40

60

80

100

69203135

5

20

36

Mus

sel s

urvi

ved

()

Juvenile

Temperature (∘C)

Salin

ities

0

20

40

60

80

100

69203135

5

20

36

Mus

sels

surv

ived

()


Temperature (∘C)

Salin

ities

0

20

40

60

80

100

69203133

5

225

40

Mus

sels

surv

ived

()

Adult

Temperature (∘C)

Salin

ities

Salin

ities

0

20

40

60

80

100

69203133

5

225

40

Mus

sels

surv

ived

()


Temperature (∘C)




418= 1653 119901 = 0205) between


218= 148361 119901 lt 0001) was





4 Discussion



Juve

nile

sur

viva

l ove

r tim

e (

)

0102030405060708090

100Cold

Mytella charruana

(Days)0 4 8 12 16 20 24 28

BC

F

CD DEE

AB AB

ABA

5 6∘C225 6∘C40 6∘C5 9∘C225 9∘C

40 9∘C5 20∘C225 20∘C40 20∘C

(a)Ju

veni

le s

urvi

val o

ver t

ime (

)

0102030405060708090

100Warm

Mytella charruana

(Days)0 4 8 12 16 20 24 28

B

A

A DA

B C

BCD

9 20∘C225 20∘C40 20∘C9 31∘C225 31∘C

40 31∘C9 33∘C225 33∘C40 33∘C

(b)


(Days)0 4 8 12 16 20 24 28

Adul

t su

rviv

al o

ver t

ime (

)

0102030405060708090

100

B

D

CD CCD

C AB

AA

5 6∘C225 6∘C40 6∘C5 9∘C225 9∘C

40 9∘C5 20∘C225 20∘C40 20∘C

(c)


Adul

t su

rviv

al o

ver t

ime (

)

0102030405060708090

100

(Days)0 4 8 12 16 20 24 28

A

A

A BA

A B

AA

5 20∘C225 20∘C40 20∘C5 31∘C225 31∘C

40 31∘C5 33∘C225 33∘C40 33∘C

(d)











Competing Interests



Acknowledgments


References














































































Volume 2014

Zoology






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology


0 45 9 18

(km)

N

W E

S

0 105 210 420

(km)


(b)

(a)








0102030405060708090

100Su

rviv

al o

ver t

ime (

)

(Days)0 3 6 9 12 15 18 21 24 27 30 33

CB

BA

6∘C

9∘C

11∘C

13∘C

(a)

Surv

ival

ove

r tim

e (

)

0102030405060708090

100

(Days)0 3 6 9 12 15 18 21 24 27 30 33

AB

C20∘C

31∘C

36∘C

(b)

Surv

ival

ove

r tim

e (

)

(Days)0 3 6 9 12 15 18 21 24 27 30 33

0102030405060708090

100

AA

BC

20∘C

23∘C

28∘C

31∘C

(c)







Juve

nile

sur

viva

l ove

r tim

e (

)

0102030405060708090

100

(Days)0 4 8 12 16 20 24 28

ColdPerna viridis

DDCDCD

ABDABA

5 6∘C20 6∘C36 6∘C5 9∘C20 9∘C

36 9∘C5 20∘C20 20∘C36 20∘C

(a)Ju

veni

le s

urvi

val o

ver t

ime (

)

0102030405060708090

100

(Days)0 4 8 12 16 20 24 28

WarmPerna viridis

DACABEFBC

CDFCDEC

5 20∘C20 20∘C36 20∘C5 31∘C20 31∘C

36 31∘C5 35∘C20 35∘C36 35∘C

(b)

(Days)0 4 8 12 16 20 24 28

Adul

t su

rviv

al o

ver t

ime (

)

0102030405060708090

100Cold

Perna viridis

C

D

CD CDCD

CD A

BAB

5 6∘C20 6∘C35 6∘C5 9∘C20 9∘C

35 9∘C5 20∘C20 20∘C35 20∘C

(c)

Adul

t su

rviv

al o

ver t

ime (

)

0102030405060708090

100

(Days)0 4 8 12 16 20 24 28

WarmPerna viridis

CDBC

AA

B

DE

EF

C

5 20∘C20 20∘C37 20∘C5 31∘C20 31∘C

37 31∘C5 35∘C20 35∘C37 35∘C

(d)












3 Results


312= 398 119901 =



212= 25153 119901 lt 00001

Exp C F39






418= 2132 119901 = 0119 Table 3) However









418= 3480





Juvenile Cold




Juvenile Warm



Adult Cold



Adult Warm











418= 2605 119901 = 0071


218

= 63372 119901 lt 0001 F218


418= 0914 119901 =






0

20

40

60

80

100

69203135

5

20

36

Mus

sel s

urvi

ved

()

Juvenile

Temperature (∘C)

Salin

ities

0

20

40

60

80

100

69203135

5

20

36

Mus

sels

surv

ived

()


Temperature (∘C)

Salin

ities

0

20

40

60

80

100

69203133

5

225

40

Mus

sels

surv

ived

()

Adult

Temperature (∘C)

Salin

ities

Salin

ities

0

20

40

60

80

100

69203133

5

225

40

Mus

sels

surv

ived

()


Temperature (∘C)




418= 1653 119901 = 0205) between


218= 148361 119901 lt 0001) was





4 Discussion



Juve

nile

sur

viva

l ove

r tim

e (

)

0102030405060708090

100Cold

Mytella charruana

(Days)0 4 8 12 16 20 24 28

BC

F

CD DEE

AB AB

ABA

5 6∘C225 6∘C40 6∘C5 9∘C225 9∘C

40 9∘C5 20∘C225 20∘C40 20∘C

(a)Ju

veni

le s

urvi

val o

ver t

ime (

)

0102030405060708090

100Warm

Mytella charruana

(Days)0 4 8 12 16 20 24 28

B

A

A DA

B C

BCD

9 20∘C225 20∘C40 20∘C9 31∘C225 31∘C

40 31∘C9 33∘C225 33∘C40 33∘C

(b)


(Days)0 4 8 12 16 20 24 28

Adul

t su

rviv

al o

ver t

ime (

)

0102030405060708090

100

B

D

CD CCD

C AB

AA

5 6∘C225 6∘C40 6∘C5 9∘C225 9∘C

40 9∘C5 20∘C225 20∘C40 20∘C

(c)


Adul

t su

rviv

al o

ver t

ime (

)

0102030405060708090

100

(Days)0 4 8 12 16 20 24 28

A

A

A BA

A B

AA

5 20∘C225 20∘C40 20∘C5 31∘C225 31∘C

40 31∘C5 33∘C225 33∘C40 33∘C

(d)











Competing Interests



Acknowledgments


References














































































Volume 2014

Zoology






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology


0102030405060708090

100Su

rviv

al o

ver t

ime (

)

(Days)0 3 6 9 12 15 18 21 24 27 30 33

CB

BA

6∘C

9∘C

11∘C

13∘C

(a)

Surv

ival

ove

r tim

e (

)

0102030405060708090

100

(Days)0 3 6 9 12 15 18 21 24 27 30 33

AB

C20∘C

31∘C

36∘C

(b)

Surv

ival

ove

r tim

e (

)

(Days)0 3 6 9 12 15 18 21 24 27 30 33

0102030405060708090

100

AA

BC

20∘C

23∘C

28∘C

31∘C

(c)







Juve

nile

sur

viva

l ove

r tim

e (

)

0102030405060708090

100

(Days)0 4 8 12 16 20 24 28

ColdPerna viridis

DDCDCD

ABDABA

5 6∘C20 6∘C36 6∘C5 9∘C20 9∘C

36 9∘C5 20∘C20 20∘C36 20∘C

(a)Ju

veni

le s

urvi

val o

ver t

ime (

)

0102030405060708090

100

(Days)0 4 8 12 16 20 24 28

WarmPerna viridis

DACABEFBC

CDFCDEC

5 20∘C20 20∘C36 20∘C5 31∘C20 31∘C

36 31∘C5 35∘C20 35∘C36 35∘C

(b)

(Days)0 4 8 12 16 20 24 28

Adul

t su

rviv

al o

ver t

ime (

)

0102030405060708090

100Cold

Perna viridis

C

D

CD CDCD

CD A

BAB

5 6∘C20 6∘C35 6∘C5 9∘C20 9∘C

35 9∘C5 20∘C20 20∘C35 20∘C

(c)

Adul

t su

rviv

al o

ver t

ime (

)

0102030405060708090

100

(Days)0 4 8 12 16 20 24 28

WarmPerna viridis

CDBC

AA

B

DE

EF

C

5 20∘C20 20∘C37 20∘C5 31∘C20 31∘C

37 31∘C5 35∘C20 35∘C37 35∘C

(d)












3 Results


312= 398 119901 =



212= 25153 119901 lt 00001

Exp C F39






418= 2132 119901 = 0119 Table 3) However









418= 3480





Juvenile Cold




Juvenile Warm



Adult Cold



Adult Warm











418= 2605 119901 = 0071


218

= 63372 119901 lt 0001 F218


418= 0914 119901 =






0

20

40

60

80

100

69203135

5

20

36

Mus

sel s

urvi

ved

()

Juvenile

Temperature (∘C)

Salin

ities

0

20

40

60

80

100

69203135

5

20

36

Mus

sels

surv

ived

()


Temperature (∘C)

Salin

ities

0

20

40

60

80

100

69203133

5

225

40

Mus

sels

surv

ived

()

Adult

Temperature (∘C)

Salin

ities

Salin

ities

0

20

40

60

80

100

69203133

5

225

40

Mus

sels

surv

ived

()


Temperature (∘C)




418= 1653 119901 = 0205) between


218= 148361 119901 lt 0001) was





4 Discussion



Juve

nile

sur

viva

l ove

r tim

e (

)

0102030405060708090

100Cold

Mytella charruana

(Days)0 4 8 12 16 20 24 28

BC

F

CD DEE

AB AB

ABA

5 6∘C225 6∘C40 6∘C5 9∘C225 9∘C

40 9∘C5 20∘C225 20∘C40 20∘C

(a)Ju

veni

le s

urvi

val o

ver t

ime (

)

0102030405060708090

100Warm

Mytella charruana

(Days)0 4 8 12 16 20 24 28

B

A

A DA

B C

BCD

9 20∘C225 20∘C40 20∘C9 31∘C225 31∘C

40 31∘C9 33∘C225 33∘C40 33∘C

(b)


(Days)0 4 8 12 16 20 24 28

Adul

t su

rviv

al o

ver t

ime (

)

0102030405060708090

100

B

D

CD CCD

C AB

AA

5 6∘C225 6∘C40 6∘C5 9∘C225 9∘C

40 9∘C5 20∘C225 20∘C40 20∘C

(c)


Adul

t su

rviv

al o

ver t

ime (

)

0102030405060708090

100

(Days)0 4 8 12 16 20 24 28

A

A

A BA

A B

AA

5 20∘C225 20∘C40 20∘C5 31∘C225 31∘C

40 31∘C5 33∘C225 33∘C40 33∘C

(d)











Competing Interests



Acknowledgments


References














































































Volume 2014

Zoology






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology


Juve

nile

sur

viva

l ove

r tim

e (

)

0102030405060708090

100

(Days)0 4 8 12 16 20 24 28

ColdPerna viridis

DDCDCD

ABDABA

5 6∘C20 6∘C36 6∘C5 9∘C20 9∘C

36 9∘C5 20∘C20 20∘C36 20∘C

(a)Ju

veni

le s

urvi

val o

ver t

ime (

)

0102030405060708090

100

(Days)0 4 8 12 16 20 24 28

WarmPerna viridis

DACABEFBC

CDFCDEC

5 20∘C20 20∘C36 20∘C5 31∘C20 31∘C

36 31∘C5 35∘C20 35∘C36 35∘C

(b)

(Days)0 4 8 12 16 20 24 28

Adul

t su

rviv

al o

ver t

ime (

)

0102030405060708090

100Cold

Perna viridis

C

D

CD CDCD

CD A

BAB

5 6∘C20 6∘C35 6∘C5 9∘C20 9∘C

35 9∘C5 20∘C20 20∘C35 20∘C

(c)

Adul

t su

rviv

al o

ver t

ime (

)

0102030405060708090

100

(Days)0 4 8 12 16 20 24 28

WarmPerna viridis

CDBC

AA

B

DE

EF

C

5 20∘C20 20∘C37 20∘C5 31∘C20 31∘C

37 31∘C5 35∘C20 35∘C37 35∘C

(d)












3 Results


312= 398 119901 =



212= 25153 119901 lt 00001

Exp C F39






418= 2132 119901 = 0119 Table 3) However









418= 3480





Juvenile Cold




Juvenile Warm



Adult Cold



Adult Warm











418= 2605 119901 = 0071


218

= 63372 119901 lt 0001 F218


418= 0914 119901 =






0

20

40

60

80

100

69203135

5

20

36

Mus

sel s

urvi

ved

()

Juvenile

Temperature (∘C)

Salin

ities

0

20

40

60

80

100

69203135

5

20

36

Mus

sels

surv

ived

()


Temperature (∘C)

Salin

ities

0

20

40

60

80

100

69203133

5

225

40

Mus

sels

surv

ived

()

Adult

Temperature (∘C)

Salin

ities

Salin

ities

0

20

40

60

80

100

69203133

5

225

40

Mus

sels

surv

ived

()


Temperature (∘C)




418= 1653 119901 = 0205) between


218= 148361 119901 lt 0001) was





4 Discussion



Juve

nile

sur

viva

l ove

r tim

e (

)

0102030405060708090

100Cold

Mytella charruana

(Days)0 4 8 12 16 20 24 28

BC

F

CD DEE

AB AB

ABA

5 6∘C225 6∘C40 6∘C5 9∘C225 9∘C

40 9∘C5 20∘C225 20∘C40 20∘C

(a)Ju

veni

le s

urvi

val o

ver t

ime (

)

0102030405060708090

100Warm

Mytella charruana

(Days)0 4 8 12 16 20 24 28

B

A

A DA

B C

BCD

9 20∘C225 20∘C40 20∘C9 31∘C225 31∘C

40 31∘C9 33∘C225 33∘C40 33∘C

(b)


(Days)0 4 8 12 16 20 24 28

Adul

t su

rviv

al o

ver t

ime (

)

0102030405060708090

100

B

D

CD CCD

C AB

AA

5 6∘C225 6∘C40 6∘C5 9∘C225 9∘C

40 9∘C5 20∘C225 20∘C40 20∘C

(c)


Adul

t su

rviv

al o

ver t

ime (

)

0102030405060708090

100

(Days)0 4 8 12 16 20 24 28

A

A

A BA

A B

AA

5 20∘C225 20∘C40 20∘C5 31∘C225 31∘C

40 31∘C5 33∘C225 33∘C40 33∘C

(d)











Competing Interests



Acknowledgments


References














































































Volume 2014

Zoology






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology









3 Results


312= 398 119901 =



212= 25153 119901 lt 00001

Exp C F39






418= 2132 119901 = 0119 Table 3) However









418= 3480





Juvenile Cold




Juvenile Warm



Adult Cold



Adult Warm











418= 2605 119901 = 0071


218

= 63372 119901 lt 0001 F218


418= 0914 119901 =






0

20

40

60

80

100

69203135

5

20

36

Mus

sel s

urvi

ved

()

Juvenile

Temperature (∘C)

Salin

ities

0

20

40

60

80

100

69203135

5

20

36

Mus

sels

surv

ived

()


Temperature (∘C)

Salin

ities

0

20

40

60

80

100

69203133

5

225

40

Mus

sels

surv

ived

()

Adult

Temperature (∘C)

Salin

ities

Salin

ities

0

20

40

60

80

100

69203133

5

225

40

Mus

sels

surv

ived

()


Temperature (∘C)




418= 1653 119901 = 0205) between


218= 148361 119901 lt 0001) was





4 Discussion



Juve

nile

sur

viva

l ove

r tim

e (

)

0102030405060708090

100Cold

Mytella charruana

(Days)0 4 8 12 16 20 24 28

BC

F

CD DEE

AB AB

ABA

5 6∘C225 6∘C40 6∘C5 9∘C225 9∘C

40 9∘C5 20∘C225 20∘C40 20∘C

(a)Ju

veni

le s

urvi

val o

ver t

ime (

)

0102030405060708090

100Warm

Mytella charruana

(Days)0 4 8 12 16 20 24 28

B

A

A DA

B C

BCD

9 20∘C225 20∘C40 20∘C9 31∘C225 31∘C

40 31∘C9 33∘C225 33∘C40 33∘C

(b)


(Days)0 4 8 12 16 20 24 28

Adul

t su

rviv

al o

ver t

ime (

)

0102030405060708090

100

B

D

CD CCD

C AB

AA

5 6∘C225 6∘C40 6∘C5 9∘C225 9∘C

40 9∘C5 20∘C225 20∘C40 20∘C

(c)


Adul

t su

rviv

al o

ver t

ime (

)

0102030405060708090

100

(Days)0 4 8 12 16 20 24 28

A

A

A BA

A B

AA

5 20∘C225 20∘C40 20∘C5 31∘C225 31∘C

40 31∘C5 33∘C225 33∘C40 33∘C

(d)











Competing Interests



Acknowledgments


References














































































Volume 2014

Zoology






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology




Juvenile Cold




Juvenile Warm



Adult Cold



Adult Warm











418= 2605 119901 = 0071


218

= 63372 119901 lt 0001 F218


418= 0914 119901 =






0

20

40

60

80

100

69203135

5

20

36

Mus

sel s

urvi

ved

()

Juvenile

Temperature (∘C)

Salin

ities

0

20

40

60

80

100

69203135

5

20

36

Mus

sels

surv

ived

()


Temperature (∘C)

Salin

ities

0

20

40

60

80

100

69203133

5

225

40

Mus

sels

surv

ived

()

Adult

Temperature (∘C)

Salin

ities

Salin

ities

0

20

40

60

80

100

69203133

5

225

40

Mus

sels

surv

ived

()


Temperature (∘C)




418= 1653 119901 = 0205) between


218= 148361 119901 lt 0001) was





4 Discussion



Juve

nile

sur

viva

l ove

r tim

e (

)

0102030405060708090

100Cold

Mytella charruana

(Days)0 4 8 12 16 20 24 28

BC

F

CD DEE

AB AB

ABA

5 6∘C225 6∘C40 6∘C5 9∘C225 9∘C

40 9∘C5 20∘C225 20∘C40 20∘C

(a)Ju

veni

le s

urvi

val o

ver t

ime (

)

0102030405060708090

100Warm

Mytella charruana

(Days)0 4 8 12 16 20 24 28

B

A

A DA

B C

BCD

9 20∘C225 20∘C40 20∘C9 31∘C225 31∘C

40 31∘C9 33∘C225 33∘C40 33∘C

(b)


(Days)0 4 8 12 16 20 24 28

Adul

t su

rviv

al o

ver t

ime (

)

0102030405060708090

100

B

D

CD CCD

C AB

AA

5 6∘C225 6∘C40 6∘C5 9∘C225 9∘C

40 9∘C5 20∘C225 20∘C40 20∘C

(c)


Adul

t su

rviv

al o

ver t

ime (

)

0102030405060708090

100

(Days)0 4 8 12 16 20 24 28

A

A

A BA

A B

AA

5 20∘C225 20∘C40 20∘C5 31∘C225 31∘C

40 31∘C5 33∘C225 33∘C40 33∘C

(d)











Competing Interests



Acknowledgments


References














































































Volume 2014

Zoology






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology


0

20

40

60

80

100

69203135

5

20

36

Mus

sel s

urvi

ved

()

Juvenile

Temperature (∘C)

Salin

ities

0

20

40

60

80

100

69203135

5

20

36

Mus

sels

surv

ived

()


Temperature (∘C)

Salin

ities

0

20

40

60

80

100

69203133

5

225

40

Mus

sels

surv

ived

()

Adult

Temperature (∘C)

Salin

ities

Salin

ities

0

20

40

60

80

100

69203133

5

225

40

Mus

sels

surv

ived

()


Temperature (∘C)




418= 1653 119901 = 0205) between


218= 148361 119901 lt 0001) was





4 Discussion



Juve

nile

sur

viva

l ove

r tim

e (

)

0102030405060708090

100Cold

Mytella charruana

(Days)0 4 8 12 16 20 24 28

BC

F

CD DEE

AB AB

ABA

5 6∘C225 6∘C40 6∘C5 9∘C225 9∘C

40 9∘C5 20∘C225 20∘C40 20∘C

(a)Ju

veni

le s

urvi

val o

ver t

ime (

)

0102030405060708090

100Warm

Mytella charruana

(Days)0 4 8 12 16 20 24 28

B

A

A DA

B C

BCD

9 20∘C225 20∘C40 20∘C9 31∘C225 31∘C

40 31∘C9 33∘C225 33∘C40 33∘C

(b)


(Days)0 4 8 12 16 20 24 28

Adul

t su

rviv

al o

ver t

ime (

)

0102030405060708090

100

B

D

CD CCD

C AB

AA

5 6∘C225 6∘C40 6∘C5 9∘C225 9∘C

40 9∘C5 20∘C225 20∘C40 20∘C

(c)


Adul

t su

rviv

al o

ver t

ime (

)

0102030405060708090

100

(Days)0 4 8 12 16 20 24 28

A

A

A BA

A B

AA

5 20∘C225 20∘C40 20∘C5 31∘C225 31∘C

40 31∘C5 33∘C225 33∘C40 33∘C

(d)











Competing Interests



Acknowledgments


References














































































Volume 2014

Zoology






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology


Juve

nile

sur

viva

l ove

r tim

e (

)

0102030405060708090

100Cold

Mytella charruana

(Days)0 4 8 12 16 20 24 28

BC

F

CD DEE

AB AB

ABA

5 6∘C225 6∘C40 6∘C5 9∘C225 9∘C

40 9∘C5 20∘C225 20∘C40 20∘C

(a)Ju

veni

le s

urvi

val o

ver t

ime (

)

0102030405060708090

100Warm

Mytella charruana

(Days)0 4 8 12 16 20 24 28

B

A

A DA

B C

BCD

9 20∘C225 20∘C40 20∘C9 31∘C225 31∘C

40 31∘C9 33∘C225 33∘C40 33∘C

(b)


(Days)0 4 8 12 16 20 24 28

Adul

t su

rviv

al o

ver t

ime (

)

0102030405060708090

100

B

D

CD CCD

C AB

AA

5 6∘C225 6∘C40 6∘C5 9∘C225 9∘C

40 9∘C5 20∘C225 20∘C40 20∘C

(c)


Adul

t su

rviv

al o

ver t

ime (

)

0102030405060708090

100

(Days)0 4 8 12 16 20 24 28

A

A

A BA

A B

AA

5 20∘C225 20∘C40 20∘C5 31∘C225 31∘C

40 31∘C5 33∘C225 33∘C40 33∘C

(d)











Competing Interests



Acknowledgments


References














































































Volume 2014

Zoology






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology








Competing Interests



Acknowledgments


References














































































Volume 2014

Zoology






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology


Acknowledgments


References














































































Volume 2014

Zoology






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology

















































Volume 2014

Zoology






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology

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