Houseflies speaking for the conservation of natural areas: a broad sampling of Muscidae (Diptera) on coastal plains of the Pampa biome, Southern Brazil

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REVISTA

BRASILEIRA

DE

Entomologia

AJournalonInsectDiversityandEvolution

w w w . r b e n t o m o l o g i a . c o m

Biology,

Ecology

and

Diversity

Houseﬂies

speaking

for

the

conservation

of

natural

areas:

a

broad

sampling

of

Muscidae

(Diptera)

on

coastal

plains

of

the

Pampa

biome,

Southern

Brazil

Ândrio

Zafalon-Silva

a,∗

_,

_Frederico

_Dutra

_Kirst

b

_,

_Rodrigo

_Ferreira

_Krüger

c

a_Universidade_Federal_do_Paraná,_Departamento_de_Zoologia,_Curitiba,_PR,_Brazil

b_Universidade_Federal_de_Minas_Gerais,_Departamento_de_Zoologia,_Minas_Gerais,_MG,_Brazil c_Universidade_Federal_de_Pelotas,_Departamento_de_{Microbiologia}_e_{Parasitologia,}_Pelotas,_RS,_Brazil

a

r

t

i

c

l

e

i

n

f

o

Articlehistory:

Received9February2018 Accepted10September2018 Availableonline5October2018 AssociateEditor:GustavoGraciolli Keywords: Atlanticforest Diptera Ecology Guilds Grasslands Muscidae

a

b

s

t

r

a

c

t

TheBrazilianCoastalPlainofthePampaBiome(CPPB),hassufferedfragmentationcausedbyresource extractionandcattleraising.Inturn,conservationproposalsareneededtopreventtheanthropisationof Pampanaturalareas.Thefirststeptowardsconservationproposalsbyusinginsectsisfaunainventories, providingdatasupportforlegislators.Thus,weundertookaregionalandbroad-scalesamplingsurvey toinvestigatethediversityofMuscidaefliesinprotectedandnon-protectedareasofCPPB.Inaddition, wecarriedoutanecologicalguilddiversityanalysisasametricapproachofbioindication.TheMuscidae samplingresultedin6314specimens,98speciestaxain31genera.Basedondiversityestimators,our samplingrepresents70–86%ofallmuscidsofCPPB.ThehighestdiversityoccursinPelotasstreams (non-protected)andTaimEcologicalStation(ahugeprotectedarea).Despitethefacttheseareasare morediversifiedandpresentmorepredatorymuscidspeciesthanothers,invasivespeciesassociated withlivestockwereobservedatahigherlevel,providingevidenceoftheimpactoflivestockproximity toprotectedareas.BasedonbiologicalcharactersofMuscidaespeciesandecologicalguildanalysis, wewereabletoidentify:(i)highdiversityofcarnivorousspeciesassociatedwithforestedandmore preservedareasand(ii)ahighlevelofafewsaprophagousspeciesasindicatorofanthropisationprocess. Ingeneral,ourresultsrepresentasignificantsteptowardsunderstandingMuscidaeinSouthernBrazil, andwedemonstratehowthepopulationecologyofmuscidfliessupportsdatatoconservationproposals. ©2018SociedadeBrasileiradeEntomologia.PublishedbyElsevierEditoraLtda.Thisisanopen accessarticleundertheCCBY-NC-NDlicense(http://creativecommons.org/licenses/by-nc-nd/4.0/).

Introduction

Theconversionoftropicalforests,naturalﬁelds,andgrasslands intoagriculturalandurbanareasistheprimarydriverofthe biodi-versitycrisiscurrentlyobserved(Foleyetal.,2005).Approximately 43%oftheterrestrialworldsurfacehasbeendisturbed,andthe originalvegetationhasbeenconvertedinto anthropogenicnew habitats(Barnoskyetal.,2012).Tropicalcountries,suchasBrazil, followthesamepatternshowinganincreasingtrendasagricultural frontiersarestillexpanding(Ferreiraetal.,2012).Extinctionrates inthiscenturyareestimatedtobemorethandoubleincomparison topreviousones(Pereiraetal.,2010).

InthestateofRioGrandedoSul(RS),thePampabiomeis sub-jectedtohighanthropicpressure(Pillaretal.,2009),andknowledge onbiodiversity isparticularlyneglected(Overbecket al.,2007).

∗ Correspondingauthor.

E-mail:andrio.zafalon@gmail.com(Â.Zafalon-Silva).

Amongareasofbiologicalimportance,wecanhighlighttheCoastal PlainofthePampaBiome(CPPB)whichhasagreatdiversityof habi-tats(wetlands,ﬂoodplains,riparianforests,andsands)(Overbeck etal.,2007).

Fragmentation process in the CPPB mainly occurs via the replacementofnativegrasslandswithagricultureandexotic pas-tures and the destruction of wetlands by drainage or barring. Additionally,frequentfires,mischaracterisationofhabitatby live-stockovergrazingandtrampling,hunting,andbiologicalinvasions arealsosignificant(Bencke,2009;Fontanaetal.,2003;Machado et al.,2008; Mikich and Bérnils,2004).In recent years, habitat losshasintensified(Bencke,2009;Crawshawetal.,2007), influ-encingsomegroupsofbirdsandmammals(Bilencaetal.,2012; FilloyandBellocq,2006;GonzalezandMerino,2008;Krapovickas and DiGiacomo, 1998).However,knowledge abouttheimpact oninsectcommunitiesinPampabiomeisincipient,despitethe effortstosurvey thediversity ofLepidoptera(Pazet al.,2013), Coleoptera (Da Silva et al., 2013), Formicidae (Hymenoptera) (Rosado et al., 2012, 2013), and Diptera. Among studied flies

https://doi.org/10.1016/j.rbe.2018.09.002

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diversity we can list the following families: Drosophilidae (Hochmülleretal.,2010;Poppeetal.,2012,2013)Muscidae(Krüger etal.,2010),Sciomyzidae(Kirstetal.,2015),Syrphidae(Kirstetal., 2017)andTabanidae(KrügerandKrolow,2015).

The Pampa biome is a highly significant area for biodiver-sityconservation(MMA,2004), butamongtheregionsofCPPB, dataoninsectbiodiversityarescarce, withtheexception ofthe municipalityofPelotasandCapãodoLeão,forMuscidae, Calliphori-dae,andTabanidaeflies(AzevedoandKrüger,2013;Krügerand Krolow,2015;Krügeretal.,2010).Studiesonthespeciesrichness ofSciomyzidae(Kirstetal.,2015)andthediversityofSyrphidae (Kirstetal.,2017)werethefirsttoreportdataoninsectfaunain mostprotectedareasintheCPPB.

The ﬁrst step towards insect conservation proposals is to increaseknowledgeonfauna,mainlybyspeciesinventories.Thus, thisstudyinvolvedaregionalandbroadsamplingsurveyto deter-minethediversityofMuscidaeonCPPB(includingthetransition zonetotheAtlanticforestinthenorthofRS).Furthermore,this studyaimedtoestablishmeasuresofdiversity(Magurran,2004)to evaluatethesuccessofsamplinginprotectedandnon-protected areas.Alimitationoftheseestimatesisthehighdependenceon samplingeffort,mainlyinthemeasuresusingabundanceand rich-ness (Bungeand Fitzpatrick, 1993). Therefore, for conservation proposals,determinationofspeciesrichnessisasessentialasthe numberofspeciestobediscovered(Santoset al.,2010).In this regard,nonparametricestimatorsofdiversitywereusedto extrap-olatespeciesrichnessinthestudiedhabitats(GotelliandColwell, 2001;Willieetal.,2012).

TheimportanceofMuscidaetotheCPPPhabitatsisthatthis familypresentsanevolutionaryhistoryinthePampabiome,its Southernforestswereanareaofhighdiversiﬁcationofthis fam-ily(Löwenberg-NetoandCarvalho,2009), andtodayalong with theAtlanticforest,ithasmanyareasofendemism(Carvalhoetal., 2003;NiheiandCarvalho,2005).Furthermore,theMuscidaeshow excellenttaxaproperties,whichmakethemsuitablefordiversity studiesinthisbiome.Theypresentbroadoccupancy,highspecies richness (Carvalhoet al., 2005)and wide habitatrange (Ferrar, 1987;Skidmore,1985).Duetothese reasons,onecanconsider muscidsﬂiesassemblagescomprisingmanyspecieswith funda-mentalsofbioindicatortaxa. Theexpression‘bioindicator’ isan aggregatetermreferringtoallsourcesofbioticandabiotic reac-tionstoecologicalchanges.Insteadofmerelyworkingasgaugesof naturalchange,taxaareutilisedtoshowtheimpactsofsurrounding naturalchanges,orenvironmentalchanges(Parmaretal.,2016).In thissense,thefollowingmaybeexpectedfromMuscidae:the pres-enceofsynanthropicspeciesasindicatorofdisturbanceinnative forestedareas(e.g.MuscadomesticaLinnaeus,Hydrotaea Robineau-Desvoidyspecies,Stomoxys calcitrans(Linnaeus), amongothers) (Greenberg,1971;Krügeretal.,2010;Skidmore,1985; Zafalon-Silvaetal.,2014).Ontheotherhand,thepresenceofspeciesthatdo nottoleratehabitatanthropisationcouldbeanindicatorofforested “healthy”environments(e.g.CyrtoneurinaGiglio-Tos,Limnophora Robineau-Desvoidy,NeomuscinaTownsend,NeodexiopsisMalloch, amongothers)(Krügeretal.,2010;Skidmore,1985;Wernerand Pont,2006).

We can argue towardsthe ecological guilds as a bioindica-torapproachinMuscidaeastheefﬁcientlydiscretedelimitation oftrophiclevelsaremainlybasedondataoflarvalmorphology (Skidmore,1985),and bythediscussiongatheredin Rodríguez-Fernándezetal.(2006)andKrügeretal.(2010).Thissurveyaims atcomplementingtheguilddescriptionsbyanalyzingthenumber ofspeciesperguildineachregion.

Italsoaimsatansweringthefollowingquestions:(i)whichisthe diversityofMuscidaeassemblagesinprotectedandnon-protected areasofCPPB?(ii)IsitpossibletouseecologicalguildsofMuscidae asatooltocheckenvironmentalaspects?

Materialandmethods

Studysites

The forested areas surveyed includedthe Grasslands of the Pampabiome,andforestsinthenortheasternCoastalPlainofthe PampaBiome(CPPB)inthetransitionzonetotheAtlanticforestof theRioGrandedoSulstate(RS),Brazil.Thesamplingareasinthe CPPB(Table1andFig.1)werechosenaccordingtothehighpriority indicatedfortheconservationofinvertebrates(MMA,2000).The MinistryofEnvironment(MMA)indicatesareasaroundthecoastal lagoonsofthePatos,Mirim–Mangueiracomplex,andthelagoons ofNorthCoastofRS(MMA,2000).

Fig.1andTable1showthestudysites.Region1ismadeupof areasaroundthePelotasstream(points1–3),Corrientesstream (points4and 5),and Turuc¸ustream(points6and 7).Region2 contains theLamiBiologicalReserve(points8and 9),Camaquã river(point10),andtheprivatenaturalheritagereserve(RPPN) BarbaNegra(points11–14).Region3containstheTaim Ecologi-calStation(points15–21),whichisasigniﬁcantareaofnational protection.TheItapuãStatePark(points22–24),TupancyNatural MunicipalityPark(point25),JoséLutzenbergerStatePark(known asGuardhousePark,point26),andItapevaStatePark(points27 and28)makeupRegion4onthenortherncoastofCPPB.Region5 istheLagoadoPeixeNationalPark(points29–35).

Thecharacterizationsofthesampledareasandtheirprotection statusbylawaresummarisedinTable1.Foracomplete charac-terisationofallareas,includingconservationunities,thereaderis referredtoWaechter(1985,2002),IBGE(1986,2004),Mello-Filho etal.(1992),SEMA(1996,2013),MeiraandPorto(1998),ICMBIO (1999),Burger(2000),Leite(2002),CeluloseRiograndense(2012),

Venzke(2012),andWitt(2013).

Samplingdesign

TheMuscidaeflieswerecollectedinfiveregionsoftheCPPB using140Malaisetraps,modelTownes(1972)withamodified col-lectioncontainer(Duarteetal.,2010).Sevenareasweresampledin eachregion(Table1andFig.1)andfourtrapswereinstalledineach area.Thetrapsremainedinthefieldforeightdays.Ontheeighth day,collectorcontainerscarryingmaterialpreservedin70% alco-holweresenttotriageandpinninginthelaboratory.Theperiods ofcollectioncampaignswerethefollowing:(i)region1between OctoberandNovemberof2011;(ii)region2inNovemberof2011; (iii)region3inDecemberof2011;(iv)region4inJanuaryof2012; (v)region5inFebruaryof2012.

The flies were identified with specific keys of Neotropical Muscidae (Carvalhoand Couri, 2002; Costacurta and Carvalho, 2005;Costacurtaetal.,2005;CouriandCarvalho,2002;Marques andCouri,2007;Nihei,2005;NiheiandCarvalho,2007; Pereira-Colavite andCarvalho, 2012;Schüehliand Carvalho, 2005), and fromtheoriginal descriptionsof somespecies.Identificationof manyspecimenswasconfirmedbycomparingwithmaterialfrom theColeçãodeEntomologiaPadreJesusSantiagoMoure,Universidade FederaldoParaná(DZUP).

ThevouchersaredepositedatDZUPandColec¸ãodeDipterado LaboratóriodeEcologiadeParasitoseVetores,UniversidadeFederal dePelotas(LEPAV).

Dataanalysis

All analyses were performed using the statistical program R (R Development Core Team, 2015) and with the statistical packagesveganandiNEXT(Hsiehetal.,2016;Oksanenetal.,2015). Thespecieswithspecimenswhichweremuchdamagedtoidentify

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Table1

Descriptionofthe35pointssampledinﬁveregionsoftheCoastalPlainonPampaBiome(CPPB).Locality:sampledareas(fourMalaisetrapperarea)insidetheregions;LP: law-protectedarea;coordinates:centralcoordinateofthesampledarea;sitedescription:ageneraldescriptionofthelocality,includingitphytophysiognomyandgeology; climate:averagetemperatureandrelativehumidityduringthesamplingperiod.APL,Pelotasstream;ACS,Corrientesstream;ATU,Turuc¸ustream;LAMI,LamiBiological Reserve;RCMQ,Camaquãriver;RPPN,privatenaturalheritagereserveBarbaNegrafarm;TAIM,TaimEcologicalStation;ITPA,ItapuãStatePark;PMTY,TupancyNatural MunicipalityPark;PEJL,JoséLutzenbergerStatePark;PEVA,ItapevaStatePark;LPXE,LagoadoPeixeNationalPark;points1–7,region1(Oct–Nov,2011);points8–14,region 2(Nov,2011);points15–21:region3(Dec,2011);points22–28,region4(Jan,2012);points29–35:region5(Feb,2012).

Point Locality LP Latitudeandlongitude Regionaldescription

1 APL No −31.72175 −52.25422

Atlanticforestremainswithpioneervegetationandfluvialinfluence.Thegeneral classificationisasemideciduousseasonalforest.Sandbank,riparian,andgalleryforests predominate.AbundantmarshesandriverstreamsfromPatosLagoonsystem.

2 APL No −31.67118 −52.21764 3 APL No −31.62781 −52.35285 4 ACS No −31.55675 −52.14568 5 ACS No −31.56435 −52.13951 6 ATU No −31.43295 −52.12173 7 ATU No −31.49678 −52.00834 8 LAMI Yes −30.23630 −51.09601

Riparianforestswithshrubbymarshes,herbaceousmarshes,sandbanks,andwetﬁelds.

9 LAMI Yes −30.25632 −51.10126

10 RCMQ No −31.12159 −51.79237 TransitionborderbetweentheAtlanticforestandPampabiomes.Abundantriparian forests.Thegeneralclassiﬁcationisasemideciduousseasonalforest.

11 RPPN Yes −30.43135 −51.23704

Farmingforestscoveredbyabundantnon-indigenousEucalyptusspecies.Heavygrovesof nativespeciescoveringdunes,internalbeaches,andallbordersofRPPN.

12 RPPN Yes −30.41323 −51.21834

13 RPPN Yes −30.42993 −51.14240

14 RPPN Yes −30.39651 −51.12725

15 TAIM Yes −32.55568 −52.50060

Severalinternallagoonsandforestedmarshes.Thepredominantphytophysiognomyisa coastalﬁeldwithsavanna-likegrasses.Thelandscapeiscomposedofamosaicoflagoons, marshes,sandbankshrubberies,dunes,andriparianforests.Farmingandcattleraisingin thesurroundingparkarea.

16 TAIM Yes −32.53865 −52.53732 17 TAIM Yes −32.56059 −52.50975 18 TAIM Yes −32.53347 −52.52538 19 TAIM Yes −32.53845 −52.53713 20 TAIM Yes −32.59684 −52.56837 21 TAIM Yes −32.63511 −52.48655

22 ITPA Yes −30.38372 −51.02095 Severalwoodedfoothillswithherbaceousvegetationandshrubbyﬁelds.The phytophysiognomyvariesfromshrubby/woodedﬁeldstodenseforests.Themosaic landscapeiscomposedofinternallagoonbeaches,riparianforests,andmobiledunes.

23 ITPA Yes −30.38291 −51.03008

24 ITPA Yes −30.36819 −51.02620

25 PMTY Yes −29.48945 −49.84442 Psammophileforestedarea,low-heightwoodproﬁle.Highesttreesareabout10-mtall. Generalphysiognomyischaracterisedbyasmallriparianforest,threeinternallagoons, oneduneof9mheight,andurbanareassurroundingthepark.

26 PEJL Yes −29.35508 −49.73529 Denseombrophilousforestinﬂuencedbyseaproximity.Paleodunesﬁxedbydenseforest cover.Mobiledunesarecoveredonlybyherbaceousandshrubvegetation.Several marsheswithswamp-likevegetation.

27 PEVA Yes −29.37978 −49.76215

28 PEVA Yes −29.37819 −49.75909

29 LPXE Yes −31.05366 −50.80873

Highseasonalvariationofsalinityinsoil,causingseasonalpatternsinphytophysiognomy. Lagooncomplexofdevelopedriparianforests,savanna-likeﬁelds,andxerophytic shrubberiesonﬁxedpaleodunes.Farmingandcattleraisinginthesurroundingparkarea.

30 LPXE Yes −31.05613 −50.81078 31 LPXE Yes −31.06581 −50.81776 32 LPXE ? −31.21367 −50.96057 33 LPXE ? −31.21715 −50.96621 34 LPXE ? −31.21911 −50.97516 35 LPXE ? −31.43412 −51.17502

ortorecogniseasmorphospecies(e.g.Coenosiasp.,Polietinasp., Pseudoptilolepissp.)wereremovedfromdataanalyses.

Dissimilarityinthesouthernassemblagesofﬂies

Thedissimilarityofﬂycommunitiesbetweenregions(1–5)and betweenSouthernBrazilmuscidassemblages(Costacurtaetal., 2003;Krügeretal.,2010;Rodríguez-Fernándezetal.,2006;and thepresent study)weresubjectedtomultivariateanalysiswith agglomerativehierarchicalclassiﬁcationbyunweightedpair-group methodusingarithmeticaverages(UPGMA),withcompletelinkage (Borcardetal.,2011;SneathandSokal,1973).

TheUPGMAclusteringmethodwasselectedbythebest Pear-son’scorrelationsindex(Table2)forveriﬁcationbetweenregions (1–5).Theindexwasobtainedcomparingthecophenetic matri-ces of various hierarchical clustering methods. The abundance wasstandardisedbyHellinger’stransformation,anddissimilarities betweenpairsofsampleswerecalculatedusingtheBray–Curtis dissimilaritycoefﬁcient(Borcardetal.,2011).UPGMAclustering ofMuscidaeassemblagesfromstudiesinSouthernBrazilwas per-formedbyJaccardindexofdissimilarityusingthebinarymatrices,

Table2

Resultsofthecopheneticmatricestest.Thechoiceofclustermethodwasbased onthehighestscore inthePearson’scorrelationsindex.UPGMA,unweighted pair-groupmethodusingarithmeticaverages;UPGMC,unweightedpair-group methodusingcentroids;WARD,Ward’sminimumvarianceclustering;WPGMA, weightedpair-groupmethodusingarithmeticaverages;WPGMC,weighted pair-groupmethodusingcentroids;PCI,Pearson’scorrelationindex.

Clustermethod PCI

UPGMA 0.8846567

WPGMA 0.8842323

WARD 0.8827867

WPGMC 0.7437148

UPGMC 0.6925564

asnotatalltheanalysed studiescontaineddataonabundance. Thisclusterwasgeneratedbycomparingonlyspecieswithspeciﬁc nameswithmorphospecieswhicharenotconsideredinallstudies. Samplingeffort

Samplingsufﬁciency wasveriﬁed usinga species accumula-tioncurve(SAC),whichwasbuiltusingtheveganpackage,with

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10 11-14 8-9 22-24 29-35 1-7 25 26-28 15-21

Fig.1. PartialmapofSouthAmerica,withdetailsoftheCoastalPlainofPampaBiome,RioGrandedoSul(Brazil).Themarkedpointsrefertoall35collectionareas(groups offourtraps).DescriptionsofeachareaareprovidedinTable1.Stars:region1;squares:region2;pentagons:region3;circles:region4;triangle:region5.

thespecaccum function. This function produces theSAC or the numberofspeciesatagivennumberofsampledpoints(Malaise trap).Withinthespecaccumfunction,the“exact”methodwasused, whichobtainedtheexpected(mean)speciesrichness.The“exact” methodﬁndstheexpectedSACusingthemethodindependently developedbyUglandetal.(2003),Colwelletal.(2004),andKindt etal.(2006).

Toestimatespeciesrichness(S),whichisspeciﬁcforeach treat-ment,Rarefaction,andChao2wereused(Chao,1987;Chaoetal., 2009;Chiuetal.,2014;ColwellandCoddington,1994;Gotelliand Colwell,2001).Thesearenon-parametricmethods,usedto esti-mateSbasedontheincidenceofspecies(presence/absence).The individual-based rarefactioncomputes the expected number of speciesinasubsampleof“n”individualsdrawnatrandomfrom asinglerepresentativesamplefromanassemblage.Thevaluesof RarefactionsandChao2wereobtainedusingthefunction “Poolac-cum,”which estimates theextrapolatedrichness index, as well asthenumberofspeciesfortherandomordinationofsampling units.

EcologicalguildsinMuscidae

Beforedefiningtheecological guilds,it isessential to deter-minehowmuscidfliesexploreenvironmentalresources.Themost notablestudyonMuscidaebiologywasSkidmore(1985),inwhich theauthorindicatedlarvalhabitmainlybythemorphologyofthe cephalo-pharyngealskeletonandtheanalspiracles.Basedonthe anatomicalfeaturesoflarvae,wecansuggestthreedefinitionsfor thebiology ofMuscidae.The firstone isthe saprophagous lar-vae,characterisedbywell-developedsuctionmechanisms,lacking oralaccessorysclerites,andlargeanalspiracles.Thesecondone isthesaprophagouslarvaewithpredatoryfacultativebehaviour in the third instar or parasitic behaviour, as in some genera

(e.g.CharadrellaWulp,PhilornisMeinert),characterisedby well-developedoralhooks,thepresenceoforalaccessorysclerites,the presenceofasuctionmechanism,andlargeanalspiracles.Thethird istheobligatecarnivorouslarvae,characterisedbywell-developed andwell-sclerotisedoralhooksandaccessorysclerites,theabsence of suctionmechanisms,and smallanal spiracles.For additional detailsofthespecies,morphology,andsubstratesusedby imma-turesseeSkidmore(1985)andFerrar(1987).

Thehabitsofadultscouldbeidentiﬁedbythehematophagous withlabiummodiﬁedforpenetrationandsuction.Thesecondtype islickersinsaprophagousspecies,withlabellathatarenotreduced andwithamoreconsiderablelateralexpansionmembranous.The lastoneisthepredators,withreducedlabella,developedprestomal teeth,andsclerotisedprementumtopenetratetheexoskeletonof theirprey(Coelho,1997;McAlpine,1981).

Therefore,wedeﬁnedanecologicalguildasagroupofspecies which exploit the sameclass of resources in a similar manner (Root,1967).InMuscidae,werecognisefourecologicalguildsor functionalgroups (sensuBlondel,2003)combiningtrophicroles of larvae and adult stages (Krüger et al., 2010).The species of CPPBareclassiﬁedaccordingtothefollowing:saprophagouslarvae and saprophagic/hematophagousadultsspecies (SS);facultative predators/parasiticlarvaeandsaprophagousadultsspecies(SPS); predatorylarvaeandsaprophagousadultsspecies(OS):predatory larvaeandadultsspecies(PP).

Region-guilddiversityproportionality

WetestedtheinﬂuenceofregionandguildofMuscidaeonthe proportionalrichnessandproportionalabundanceusingANOVA andFtest,consideringP<0.05assigniﬁcant.

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Table3

ListofMuscidae(Insecta,Diptera)speciescollectedinﬁveregionsoftheCoastalPlainofPampaBiome(CPPB),RioGrandedoSul,Brazil.SampledCPPBareasincludeR1: region1,where:APL:Pelotasstream;ACS:Corrientesstream;ATU:Turuc¸ustream.R2:region2,where:LAMI:LamiBiologicalReserve;RCMQ:CamaquãRiver;RPPN:private naturalheritagereserveBarbaNegrafarm.R3:region3,where:TAIM:EcologicalStationofTaim.R4:region4,where:ITPA:ItapuãStatePark;PMTY:TupancyNatural MunicipalityPark;PEJL:JoséLutzenbergerStatePark;PEVA:ItapevaStatePark.R5:region5,whereLPXE:LagoadoPeixeNationalPark.

Taxa R1 R2 R3 R4 R5 (

)

APL ACS ATU RPPN RCMQ LAMI TAIM ITPA PEVA PMTY PEJL LPXE Biopyrelliabipuncta(Wiedemann,1830) 0 0 0 0 4 0 13 8 0 0 0 3 28 BithoracochaetaannulataStein,1911 0 0 0 0 0 1 0 0 0 0 0 33 34 BithoracochaetacalopusBigot,1885 120 44 98 7 43 6 160 0 1 0 0 26 505 Bithoracochaetaﬂavicoxaa_Malloch,₁₉₃₄ ₁ ₁₂ ₂ ₀ ₁₀ ₁ ₄ ₁ ₀ ₃ ₀ ₀ ₃₄

BithoracochaetaplumataAlbuquerque,1955 0 0 0 0 0 0 0 1 0 0 0 0 1

Bithoracochaetasp.2 0 0 0 0 0 0 0 1 0 0 0 6 7

Coenosiasp. 1 0 0 0 0 0 0 1 0 0 0 0 2

Coenosiasp.1 1 0 0 0 0 0 1 0 0 0 0 0 2

CoenosiiniGenusA 3 0 0 0 0 0 2 0 0 0 0 0 5

CoenosiiniGenusB 0 0 0 0 0 0 1 0 0 0 0 0 1

Cyrtoneurinacostalis(Walker,1853) 5 3 0 0 10 0 2 2 0 0 0 1 23

Cyrtoneuropsisbrunnea(Hough,1900) 1 0 0 0 2 1 1 0 0 0 0 1 6

Cyrtoneuropsismaculipennisa_(Macquart,₁₈₄₃₎ ₁₀ ₄ ₄ ₁₁ ₀ ₁₀ ₀ ₀ ₀ ₁ ₁₁ ₀ ₅₁

Cyrtoneuropsispararescita(Couri,1995) 0 0 0 0 0 0 1 0 0 0 0 9 10

Cyrtoneuropsisincognitaa_(Snyder,₁₉₅₄₎ ₁ ₀ ₀ ₂ ₀ ₁₇ ₀ ₁₈ ₄ ₀ ₀ ₀ ₄₂

Cyrtoneuropsissp.2 0 0 0 2 0 6 0 0 0 0 0 0 8

Dolichophaoniacacheutaa_(Snyder,₁₉₅₇₎ ₀ ₀ ₀ ₀ ₀ ₁ ₀ ₀ ₀ ₀ ₀ ₀ ₁

Dolichophaoniaplaumanni(Carvalho,1983) 0 0 0 0 0 0 0 0 0 0 0 1 1

Dolichophaoniatrigona(ShannonandDelPonte,1926) 0 5 1 0 0 0 1 1 0 0 0 0 8

Dolichophaoniaelongataa_{(Albuquerque,}₁₉₅₈₎ ₀ ₀ ₀ ₀ ₀ ₀ ₁ ₀ ₀ ₀ ₀ ₁ ₂

GraphomyaauricepsMalloch,1934 2 0 1 0 0 0 21 0 0 0 0 0 24

Gymnodiadelecta(Wulp,1896) 0 0 0 0 0 0 15 0 0 0 0 0 15

Gymnodiaquadristigma(Thomson,1869) 0 0 0 0 0 0 9 1 0 0 0 0 10

Gymnodiasp. 0 0 0 0 0 0 1 0 0 0 0 0 1

Haematobiairritans(Linnaeus,1758) 0 0 0 0 0 0 20 0 0 0 0 11 31

Helinasp.1 1 5 2 0 0 0 2 0 0 0 0 0 10

Helinasp.2 26 4 3 0 2 0 6 0 0 0 0 0 41

Helinasp.3 8 0 1 0 0 1 4 1 0 1 0 0 16

Helinasp.4 0 6 1 0 0 0 4 0 0 0 0 0 11

Helinasp.6 2 5 1 5 0 4 1 5 0 0 0 24 47

LimnophoraaurifaciesStein,1911 14 7 7 3 5 15 19 0 0 0 0 0 70

Limnophorasp.1 4 0 0 0 1 1 12 0 0 0 0 1 19 Limnophorasp.2 13 2 2 0 1 0 50 1 0 0 0 0 69 Limnophorasp.3 3 0 0 0 0 0 7 0 0 0 0 0 10 Limnophorasp.4 99 11 8 6 5 5 155 2 0 1 4 2 298 Limnophorasp.5 59 36 22 8 9 3 216 1 1 0 1 0 356 Limnophorasp.6 0 1 0 0 0 0 2 0 0 0 0 0 3

LispeserotinaWulp,1896 1 0 0 0 0 2 0 0 0 0 0 0 3

Micropotamiaminuscula(Albuquerque,1955) 7 8 0 2 0 0 16 0 0 0 1 0 34

Morelliahumeralis(Stein,1918) 0 0 0 0 0 0 2 17 0 0 0 0 19

MorelliapaulistensisPamplonaandMendes,1995 0 0 0 8 58 5 40 6 1 0 0 1 119

Morelliasp.1 0 2 1 0 8 0 3 0 0 0 0 0 14

Morelliasp.2 1 1 0 1 2 0 1 0 0 0 0 0 6

MuscadomesticaLinnaeus,1758 0 0 0 0 0 0 0 0 0 1 0 0 1

MydaeaplaumanniSnyder,1941 29 20 5 2 3 0 1 2 0 0 0 0 62

Myospilaobscura(ShannonandDelPonte,1926) 1 7 0 1 6 0 13 5 1 0 2 0 33

Myospilasp.1 4 1 0 0 0 0 4 0 0 1 1 1 12

NeodexiopsiserectaCostacurta,CouriandCarvalho,2005 0 0 0 0 0 0 0 0 0 1 0 0 1

NeodexiopsisrusticaAlbuquerque,1956 2 20 0 1 0 0 1 0 0 0 0 1 25

Neodexiopsissp. 0 0 0 0 0 0 0 1 0 0 0 0 1 Neodexiopsissp.1 3 2 0 0 0 0 75 0 0 0 0 0 80 Neodexiopsissp.2 0 1 0 0 2 0 2 0 0 0 0 0 5 Neodexiopsissp.3 1 0 4 0 0 1 108 0 0 0 1 0 115 Neodexiopsissp.4 98 29 79 0 1 1 101 0 0 0 0 0 309 Neodexiopsissp.5 2 13 3 5 0 0 18 0 1 0 0 0 42 Neodexiopsissp.6 0 0 0 0 0 0 1 0 0 0 0 0 1 Neodexiopsissp.7 28 14 5 6 9 3 7 5 0 0 1 0 78 Neodexiopsissp.8 0 0 0 0 0 0 2 1 0 0 0 0 3 Neodexiopsissp.9 154 88 86 4 31 8 259 3 5 0 2 4 644 Neodexiopsissp.10 0 0 0 0 0 3 1 0 0 0 0 0 4 Neodexiopsissp.11 12 5 1 0 3 2 0 0 0 0 0 4 27

Neodexiopsissp.12affvulgarisa_Couri_and_Albuquerque,₁₉₇₉ ₀ ₀ ₀ ₁ ₀ ₀ ₀ ₀ ₀ ₀ ₀ ₀ ₁

Neodexiopsissp.13 0 0 1 0 0 0 0 0 0 0 0 0 1

Neomuscinaaffdouradensisa_Lopes_and_Khouri,₁₉₉₆ ₁ ₀ ₀ ₀ ₀ ₀ ₀ ₀ ₀ ₀ ₀ ₀ ₁ Neomuscinaaff.Goianensisa_Lopes_and_Khouri,₁₉₉₅ ₀ ₀ ₀ ₀ ₁ ₀ ₀ ₀ ₀ ₀ ₀ ₀ ₁

Neomuscinainﬂexa(Stein,1918) 0 0 0 0 0 0 0 0 0 0 0 1 1

NeomuscinaneosimilisSnyder,1949 0 0 0 0 0 0 0 1 0 0 3 0 4

Neomuscinapictipennispictipennis(Bigot,1878) 1 0 0 0 2 0 0 2 1 0 4 0 10

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Table3(Continued)

Taxa R1 R2 R3 R4 R5 (

)

APL ACS ATU RPPN RCMQ LAMI TAIM ITPA PEVA PMTY PEJL LPXE Neomuscinazosteris(ShannonandDelPonte,1926) 33 89 4 0 4 0 9 1 0 0 0 4 144

Neomuscinasp.1 0 0 0 1 0 0 0 0 0 0 0 0 1

Neomuscinasp.2 2 1 0 1 0 0 0 0 0 0 0 0 4

Neomuscinasp.3 1 0 0 1 1 0 0 0 0 0 0 0 3

Neomuscinasp.4 1 1 0 0 0 0 0 0 0 0 0 0 2

Neomuscinasp.5 0 0 0 0 0 0 0 1 1 0 1 0 3

Neurotrixafelsina(Walker,1849) 3 0 0 0 1 0 21 0 0 0 0 0 25

Neurotrixamarinoniia_Costacurta_and_Carvalho,₂₀₀₅ ₁ ₉ ₂ ₀ ₀ ₀ ₅ ₂ ₀ ₀ ₀ ₄ ₂₃

NeurotrixasulinaCostacurtaandCarvalho,2005 1 3 0 1 0 0 8 1 0 0 0 0 14

PhaoniaadvenaSnyder,1957 0 0 0 4 0 0 0 3 2 1 0 0 10

Phaoniaannulata(Albuquerque,1957) 9 21 0 0 0 0 0 4 0 0 1 0 35

Phaoniabigotia_{(Albuquerque,}₁₉₅₇₎ ₀ ₀ ₀ ₀ ₀ ₀ ₀ ₁ ₀ ₀ ₀ ₀ ₁

Phaoniagrajauensis(Albuquerque,1957) 6 13 1 10 2 7 1 10 2 0 5 0 57

Phaonianigriventris(Albuquerque,1954) 18 18 0 8 3 1 160 20 1 0 2 69 300

Phaoniapraesuturalis(Stein,1904) 0 0 0 0 2 0 0 9 0 0 0 0 11

Phaoniasimilata(Albuquerque,1957) 1 0 0 8 4 3 1 3 2 0 3 1 26

Phaoniatrispila(Bigot,1885) 7 20 18 2 50 0 64 0 0 0 0 7 168

Phaoniasp.1 8 10 1 2 5 5 24 5 11 2 10 9 92

Phaoniasp.2 0 4 1 1 0 3 316 0 0 0 0 13 338

Phaoniasp.3 22 0 6 1 9 2 0 0 0 0 0 0 40

Phaoniasp.4 0 0 0 0 0 1 0 0 0 0 0 0 1

Philornisseguyia_Garcia,₁₉₅₂ ₁ ₀ ₀ ₀ ₀ ₀ ₀ ₀ ₀ ₀ ₀ ₀ ₁

Philornissp.2 0 0 0 0 1 0 0 0 0 0 0 0 1

Plumispinasp.1 0 0 0 0 0 0 1 0 0 0 0 0 1

Polietinaorbitalis(Stein,1904) 21 16 5 3 9 0 16 4 2 0 3 17 96

Pseudoptilolepisnudapleuraa_Snyder,₁₉₄₉ ₀ ₀ ₀ ₅ ₀ ₀ ₀ ₀ ₀ ₀ ₀ ₀ ₅

Pseudoptilolepissp. 2 0 1 0 0 0 0 0 0 0 0 0 3

Sarcopromuscapruna(ShannonandDelPonte,1926) 5 2 3 0 43 1 903 0 0 0 0 18 975

Stomopogonargentina(Snyder,1957) 10 30 3 0 1 1 398 0 0 0 0 0 443

Stomoxyscalcitrans(Linnaeus,1758) 0 0 0 0 0 0 2 0 0 0 0 3 5

Trichomorelliaseguyia_(Pamplona,₁₉₈₃₎ ₀ ₀ ₀ ₀ ₁ ₀ ₂₂ ₀ ₀ ₀ ₀ ₀ ₂₃

(

) 871 593 383 123 355 121 3337 151 36 12 56 276 6314

a_New_record_for_RS_state.

Results

MuscidaeoftheCoastalPlainofPampaBiome

We collected 6314 specimens, 98 species in 31 genera of Muscidae on the CPPB (Table 3). The most abundant species were Sarcopromusca pruna (Shannon and Del Ponte) (n=934), Neodexiopsis sp. 9 (n=641), Bithoracochaeta calopus (Bigot) (n=503), Stomopogon argentina (Snyder) (n=388), Limnophora sp.5 (356),Phaonia sp. 2(n=338), Neodexiopsissp. 4 (n=309), Phaonia nigriventris (Albuquerque) (n=300), Limnophora sp. 4 (n=298).

Amongthe 98 identifiedspecies of Muscidae,41 didnot fit descriptionsofvalidspecies,indicatingthattheremaybemany newspeciesinthestudiedregions.Otherdatashowthat34taxa wereuniquetoatleastonelocation(Table3).Thegeographical distributionof13 species ofMuscidaewasexpanded, and they wereregisteredforthefirsttimeinthestateofRioGrandedoSul, basedonthedistributionalcatalogueofSouthAmericanmuscids (Löwenberg-NetoandCarvalho,2013).

Regardingthenumberofspeciesasafunctionofthesamples (areas)inCPPB,theSACexhibitedanupwardtrend(Fig.2),although theChao2diversityestimatorreportwascollectedbetween70% and86%MuscidaeofCPPB(Table4).

ThediversityofmuscidﬂiesthroughCPPBregionsand dissimilaritywithSouthernBrazilfauna

Thespeciesrichnesswasthehighestinregions1and3,where therewasagreaterabundanceofﬂies.Whenrarefactionbyregion was applied, computing the expected number of species in a

subsample ofnindividuals(n=255,correspondingtoregion4), regions2and4hadthehighestestimatedrichness(Table4). Rar-efactionbyareawascomputedtoasubsampleofn=8individuals, andthehighestrareﬁedrichnesswasobservedinareasofregions 2,4,and1(Table4).

Themostabundantspecieswerefoundtovaryineachregion. Inregion1,themostabundantspecies wereNeodexiopsissp.9, Neodexiopsissp.4,andB.calopus,whicharesmallpredatorsin lar-vaeandadultstages,andtheircombinedabundancerepresented 38%ofthecollectedspecimensfromthisregion.Singletonsand doubletonsaccountedfor 31.9%ofthetotal speciesgatheredin thisregion.Inregion2,themostabundantspecies(28.9%)were MorelliapaulistensisPamplonaandMendes,Phaoniatrispila(Bigot), and B.calopus. Inthis region,44.4%of thespecieswere single-tonsand doubletons. More than52% of thetotal abundanceof speciesintheCPPBwasinregion3(TaimEcologicalStation).In this region,46.8% of thespecimens wereS. pruna,S. argentina, andPhaoniasp.2.Wecollected26species(36.62%)ofsingleton anddoubletonsinthisregion.Inregion4,Phaoniasp.1,Phaonia nigriventris (Albuquerque),and Cyrtoneuropsis incognitaMalloch werethemostabundantspecies(28.6%).Region4contained48.9% ofsingletonand doubletonspecies.Inregion5 (Lagoado Peixe NationalPark),themostabundantspecies,correspondingto55.5%, wereP.nigriventris,B.calopus,B.annulataSteinandHelinasp.6. Singletonanddoubletonspeciesaccountedfor40%ofthespecies richness(Table3).

Differencesinspeciescompositionbetweenregionsareshown by the cluster analysis(Fig.3).The ﬁrst cluster group is com-posedbyregions1,2,and3.Theassemblagedissimilaritybetween groups from regions 1 and 2 was more than 40%. The differ-encebetweentheclusterformedbyregions1and2andformed

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60 50 40 30 20 10 0 60 50 40 40 30 30 20 20 10 10 0 0 30 100 80 60 40 20 0 25 20 15 10 5 0 60 50 40 30 20 10 0 1 2 3 4 5 6 7 1 2 3 4 5 6 7 1 2 3 4 5 6 7 1 2 3 4 5 6 7 0 5 10 15 20 25 30 35 1 2 3 4 5 6 7 Number of samples Number of samples

Number of samples Number of samples

Number of species

Number of species Number of species

Number of species

a

_b

c

d

e

f

Fig.2.SamplecoveragecurvesbasedonthenumberofindividualsofMuscidaeperregionintheCoastalPlain(CPPB)ofRioGrandedoSul,Brazil.Thegreyshadedarea representstheconﬁdenceintervals.(a)region1;(b)region2;(c)region3;(d)region4;(e)region5;(f)CPPBtotal.

Cluster dendrogram Region 5 Region 4 Region 3 Region 1 Region 2 Dissimilar ity 0.65 0.65 0.55 0.50 0.45 0.40

Fig.3.CompositionaldifferencesbetweenMuscidae(Insecta,Diptera)assemblagesofallﬁveregionsoftheCoastalPlainofPampaBiome(RioGrandedoSul,Brazil)shown bytheunweightedpair-groupmethodusingarithmeticaverages(UPGMA)clusteranalysis.

byregion3wasalsolow,atabout45%.Moresigniﬁcant differ-encesbetweenassemblageclusterswereobservedbetweengroup 1(regions1–3)andregions4and5,withmorethan60% dissimi-larity.AmongthespeciesofmuscidsinSouthernBrazil,including thoseinthisstudy,therewashighdissimilaritybetweenclusters witha trendtoincrease/decreasebasedongeographicdistance (Fig.4).

DiversitypatternbytrophicguildsofMuscidae

Theproportionofspeciesandabundancebyguildwasnot con-sistent throughout theCPPB; this difference wasinﬂuenced by collectionregion(Tables5and6).Intheﬁrstthree regions,the highestspeciesrichnesswasfoundinaguildofPPsettingsfollowed byOSsettings;intheformer,bothlarvaeandadultsarepredators,

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Table4

Richnessestimatorsbylocationandregionofcollection.Nonidentiﬁedspecimenswereprunedfromdatamatricesbeforeanalysis.R1:region1,where:APL:Pelotasstream; ACS:Corrientesstream;ATU:Turuc¸ustream.R2:region2,where:LAMI:LamiBiologicalReserve;RCMQ:Camaquãriver;RPPN:privatenaturalheritagereserveBarbaNegra farm.R3:region3,where:TAIM:TaimEcologicalStation.R4:region4,where:ITPA:ItapuãStatePark;PMTY:TupancyMunicipalityNaturalPark;PEJL;JoséLutzenberger StatePark;PEVA:ItapevaStatePark.R5:region5,whereLPXE:LagoadoPeixeNationalPark.CPPB,CoastalPlainsonPampaBiome;Traps,numberoftrapsperlocation;S, speciesrichness;Abund,abundance;Chao2,Chao2non-parametricdiversityestimator;Srar,rarefaction;SE,standarderror.Intherarefactiondatabylocation,thestandard numberofindividualswas12(PMTY),andbyregionthestandardnumberofindividualswas254(R4).

Traps S Abund Chao2±SE Srar±SE

R1 APL 12 55 870 81.96±17.6 8.08±1.36 ACS 8 42 592 46.49±4.79 8.80±1.34 ATU 8 34 383 51.95±14.35 6.22±1.35 TotalR1 28 62 1845 104.23±38.5 38.40±2.65 R2 LAMI 8 31 121 54.8±20.03 9.07±1.27 RCMQ 4 39 355 46.12±5.88 8.04±1.36 RPPN 16 32 123 40.26±6.85 9.62±1.17 TotalR2 28 60 599 72.78±8.36 46.15±2.60 R3 TAIM 28 62 3333 74.5±8.46 30.11±2.59 R4 ITPA 12 37 150 72.88±25.7 9.01±1.30 PMTY 4 9 12 31.45±28.54 9.00 PEVA 8 15 36 22.77±7.29 7.62±1.26 PEJL 4 18 56 26±8.13 8.08±1.26 TotalR4 28 46 254 86.34±28.44 46 R5 LPXE 28 29 276 78.82±59.37 28.19±0.84 CPPBtotal 140 98 6307 181.65±58.58 – 50 55 60 65 70 75 Dissimilarity %

Fig.4.TheUPGMAclusterbetweenMuscidae(Insecta,Diptera)assemblagesofSouthernBrazil.ThedissimilarityanalysiswasperformedusingtheJaccardindexwithbinary matrices.InParanástate:greendiamonds,Rodríguez-Fernándezetal.(2006);pinkcircles,Costacurtaetal.(2003).IntheRioGrandedoSulstate:redstars,currentstudy; bluesquares,Krügeretal.(2010).

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25 400 300 200 100 0 20 15 10 5 0 1 2 3 4 5 1 2 3 4 5 Richness Ab undance OS PP SPS SS

a

b

Fig.5.Graphicalrepresentationofproportionalrichness(a)andabundance(b)bytheguildintheﬁveregionsofCoastalPlainofPampaBiome(RioGrandedoSul,Brazil).SS, saprophagouslarvaeandsaprophagic/hematophagousadults;SPS,facultativepredators/parasiticlarvaeandsaprophagousadults;OS,predatorylarvaeandsaprophagous adults;PP,predatorylarvaeandadults.

Table5

Analysisofvariance(ANOVA)andFtest,testingtheinﬂuenceofregionandecological guildsofMuscidaeontheproportionalrichness.Df,degreesoffreedom;SumSq, sumofsquares;MeanSq,meanofsquares.

Df SumSq MeanSq Fvalue P Guild 3 1.51893 0.50631 61.9787 <0.001 Region 4 0.00010 0.00002 0.0030 1.000 Guild:region 12 0.51359 0.04280 5.2392 <0.001 Residuals 120 0.98029 0.00817 – –

Table6

Analysisofvariance(ANOVA)andFtest,testingtheinﬂuenceofregionandecological guildsofMuscidaeontheproportionalabundance.Df,degreesoffreedom;SumSq: sumofsquares;MeanSq:meanofsquares.

Df SumSq MeanSq Fvalue Pr(>F) Guild 3 1.8376 0.61254 21.9057 <0.001 Region 4 0.0048 0.00119 0.0426 0.997 Guild:region 12 2.0160 0.16800 6.0079 <0.001 Residuals 120 3.3555 0.02796 – –

andinthelatter,larvaearepredatorsandadultsaresaprophagous (Fig.5A).Inregions4and5,thisrelationshipwasreversed,witha predominanceofOSsettingsspeciesfollowedbyPPguild(Fig.5A). Inallregions,guildspecieswithsaprophagoushabitsinthelarval andadultstagesispresentinthelowestproportionofthetotal fauna,exceptinregion3(Taim),whichexhibitedincreased rich-nesscomparedwiththeSPSguild(Fig.5A).Thesamepatternwas observedfortherelativeabundancewithineachguildbyregion. Contrarytotherelationshipbetweentheproportionofspeciesper guildandregion,theabundanceoftheSSguildgreatlycontributes tothefaunainregions3and5(Fig.5B).

Discussion

TheMuscidaeinSouthernBrazil,faunasimilarity,andpossible areasofendemism

InSouthernBrazil,majortaxonomicsurveysofMuscidaewere carriedout,andatrendofgreaterorlessersimilaritybetweenthe areasofcollectionswasobservedwithincreasingordecreasing dis-tance.Therewasgreatersimilaritybetweenthedatacollectedin CPPBandthoseinventoriedbyKrügeretal.(2010)intheRioGrande doSulovera1-yearcollectionperiod.Weshare36specieswithout

themorphospecies,andtherewasatendencyofsimilarityamong themostabundantspeciesinthetwostudies.Surveysconducted intheParanástate(Costacurtaetal.,2003;Rodríguez-Fernández etal.,2006),whichisabout1000kmfromthestudysiteinthe CPPB,foundthattheproportionofspecieswaslower,with17and 23species,andthereisahighdissimilaritywithCPPB(Fig.4).Some studies(Löwenberg-NetoandCarvalho,2009;NiheiandCarvalho, 2005)have indicated areasofendemism and diversiﬁcationfor MuscidaeintheParanaenseForest(Paranástate)andthePampa. Therefore,ourdataindicatethattheCPPBregionsmayrepresent newareasofendemism,sincethereis:(i)signiﬁcantMuscidae rich-ness;(ii)newrecordsofoccurrence;(iii)highratesofparticular speciesperregion;(iv)highdissimilaritieswiththeParanáForest communities.

SamplingeffortintheareasandconservationunitsintheCPPB DifferencesbetweenthesamplingsbyCostacurtaetal.(2003),

Rodríguez-Fernándezetal.(2006),andKrügeret al.(2010)and thoseinthepresentstudyincludetheexposuretimeofMalaise trapsversus thenumber oftraps installed byarea. Thestudies byCostacurtaetal.(2003),Rodríguez-Fernándezetal.(2006),and

Krügeretal.(2010)addressedthesamplingofMuscidaeassemblies onatemporalscaleattheexpenseofaspatialscale.Incontrast,we soughttodirectlysampleonaspatialscaleovertime,withmany replicates(traps)andashortperiodofexposure.

Whilerepresentingaconsiderablesamplingeffort(140Malaise trapsin35areas),ourspeciesaccumulationcurvepossesses no asymptote.We can argue that (i) therestricted useof Malaise trapsisnotproblematicforthesamplingofMuscidaediversity,as oncecomparedwithtrapscontainingdecomposingbait(Carvalho etal.,1984),theMalaisetraphashigherefﬁciencyinthecapture ofDipterarichness(Brown,2005;Costacurtaetal.,2003;Duarte etal.,2010;Krügeretal.,2010;Rodríguez-Fernándezetal.,2006). Speciescollectedwithbaitsareoften,thoughnotalways,gathered inMalaisetraps;(ii)theascendingpatternoftheSACshowsthat morefocusshouldbeplacedontrapsinthelargefragmentsthanin thesmallfragments.Itispossiblethattherichnesshasbeen under-estimatedinlargerareasasthecollectioneffortwasthesamein largeandsmallareas.Therefore,smallareasshowedmorerichness intheCPPB(e.g.,PelotasandCorrientesstreams,Camaquãriver, TaimecologicalStation,Itapuãstatepark).(iii)Inregions4and5, anincreaseinthenumberoftrapspersecannotreversetheupward trendoftheSAC.Insuchareas,itisnecessarytomovethecollection

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periodbeforeJanuary(thebeginningofsummerintheSouthern Hemisphere)duetoclimaticconditionsofhightemperaturesand lowrelativehumidity.Theseconditionshaveprobablyprevented themovementofadultspecimens,orelsecausedtheirdeath, pre-ventingthemfrombeingcollectedbyMalaisetraps.Despitethis fact,analysiswithChao2revealedthat75%ofMuscidaespecies werecollectedinsomeregions(Table4),suchastheareasaround thePelotas,Corrientes,andTuruc¸ustreams.Therewasmore rich-nessinregion1thanattheTaimEcologicalStation(Tables3and4). Theareasinregion1arenotprotectedandsufferhighexploitation duetoextractivepracticeandagriculturalproductionpracticesin theirsurroundings.

Ingeneral,singletonsanddoubletonsspeciesrepresent29.7% oftherichness,andtheirproportionrangesfrom31.9to48.9%of thespeciescollectedindifferentregions,whichisatypicalpattern forinsectcommunitiesintropicalforests(Erwin,1998;Bassetand Kitching,1991;Novotn ´y,1993).

Muscidaeecologicalguildsasindicatorsofpriorityareasfor conservation

Ecological/trophic guilds of Muscidae can beused to assess environmental health as theyrespond to specific features. For example,oneofthefactorscloselylinkedtotheconservationof natural areasis theabundanceof predatoryspecies of Neodex-iopsisandLimnophora.Thespeciesofthesegeneraoccurinareas wherethereishighavailabilityoffreshwatersourcesforthe devel-opmentof larvae,and a highabundanceof preyfor thelarvae andadultsof thesespecies (WernerandPont, 2006).Thepreys recordedintheliteratureincludechironomids,simulids,andother smallfliesthatdonottoleratedisturbedareas(WernerandPont, 2006).ThePPguild,whichincludesthepredators,wasdominant inmostareas,especiallyinregions1–3(Fig.4).Intheseregions, therearemanymarshes,smallrivers,streams,andotherflooded areas,whicharethesurroundingphysiognomyofthePatoslagoon andMirim–Mangueiralagoon,andpriorityareasforconservation accordingtoMMA(2000).Therefore,thehighrichnessand abun-dance proportional to thePP guild indicate potential areas for permanentprotection,suchasthePelotasstreams(Pelotas, Cor-rientes,Turuçu)andtheCamaquãriver.

Otherguildsmaybeindicationsofadisturbanceinnaturalareas, specificallytheSSguild,containinglarvaeandsaprophagousadults. ThespeciesintheSSguilddevelopedpreferentiallyinwildand domesticanimalfaeces;therefore,thiswouldbeexpecteda pri-oriforhighdiversity andabundancein TaimEcological Station, whichisarefugeforawidevarietyofbirds,reptiles,and mam-mals.ProportionalityanalysisshowedthattheSSguildcontainsa higherabundanceinregion3.Thisresultisnotinconsistentwith expectations,asthedominantspeciesinthisguildisSarcopromusca pruna.Thelarvaeofthisspeciesareassociatedwithanimalfaeces (Pedroso-De-Paiva,1996),whileadultsfeedonwoundsinduced bybloodsuckingspeciessuchashorseflies,hornflies (Haemato-biairritans),and stableflies(Stomoxyscalcitrans). Thesespecies arecommonlyassociatedwithandrecognisedasvectorsofmany pathogensandeggsofparasites(Azevedoetal.,2007).The biol-ogyofthehornflyshouldbenoted,wherebytheadultconstantly remainsonthehosttofeed,andfemalesmoveawayfromthehost onlyforovipositiononnewlydepositedfaeces,astheyare depen-dentonhighhumidityindungpats(McLintockandDepner,1954). ThispeculiarbehaviourshouldlimittheircaptureinMalaisetraps sinceH.irritansdo notoftenfly, confirmingthetrafficofcattle withinfragmentsofTaim.ThepresenceofspeciesoftheSSguild, S.pruna,H.irritans,and S.calcitrans,alongwiththeoccurrence ofGymnodiaspecies(OSguild),whichalsooccursinthestoolsof productionungulates,areanindicationofdisturbance.

The OS guild containingpredatory larvaeand saprophagous adultsgenerallycontainsgeneralistpredatorylarvaeintheir micro-habitats (Phaonia, Helina, Graphomya), and often the faeces of herbivorousvertebrates(Gymnodia)(Skidmore,1985).The pres-enceofspeciesofthisgenusindicatesadisturbanceinthenatural environmentduetotheassociationwithcattledung,unlike indige-nousspeciesofPhaonia,Helina,andGraphomya,wherethelarvae arepredatorsinhumusandmosses.Thehighproportionofthe OSguildinLagoadoPeixe(region5),withouttheoccurrenceof Gymnodiaspecies,isindicativeofareasofdiverseresources, con-sistentlowanthropisedarea.

Closingremarks

OurresultsshowingeneralthatsomehabitatsofCPPBare suf-feringfrominvasionsbynon-indigenousspeciescloselyassociated with livestock. The invasions present a danger to the indige-nous species which occupy the same space as these invaders, suchasherbivorousfaeces.Thistopicraisesdiscussionaboutthe implementation of buffer zones and monitoring the anthropic activitiessurroundingprotectedareas(Allanetal.,2017;Weisse andNaughton-Treves,2016).

Conversely, the non-protected areas in Pelotas streams and CamaquãRiverneedmorelegalattentionbylegislatorstobecome protected areas due to the high diversity of ﬂies and high proportionalityofguildsofpredators(indicatorsforhealthy envi-ronments).DespiteoursamplingeffortsinCPPB,theresultsreveal theneedforadditionalsurveysin theseregions,asmanymore speciesremaintobediscovered.TheCPPBmaybeconsidereda hotspot for theconservationof Muscidae species,in turn, new biogeographicalapproaches areneededtoidentifynewareasof endemisminthePampabiome.

Conﬂictsofinterest

Theauthorsdeclarenoconﬂictsofinterest.

Acknowledgements

Allauthorshavecontributedsubstantially totheconception, design,analysis,and/orinterpretationofthedatainthismanuscript andwilltakepublicresponsibilityforthecontent.AZSwas sup-portedbyCoordenaçãodeAperfeiçoamentodePessoaldeNível Superior(CAPES)andbytheNationalCouncilofTechnologicaland ScientificDevelopment(CNPq)(processno.161907/2015-6).We alsothankCNPqforfinancingtheproject“Efeitodareduçãodaárea nadiversidadedeMuscidae(Insecta:Diptera)daplaníciecosteira do Rio Grande do Sul” (processno.473949/2010-5). We thank JulianoLessaPintoDuarte,PhD,forassistanceinthegenus sort-ingofmaterial;ClaudioJoséBarrosdeCarvalho,PhD(Universidade FederaldoParaná)forprovidinghelpwithidentification.Wealso thanktheadministratorsofprotectedareaswherecollectingwas conductedandto“SistemadeAutorizaçãoeInformaçãoem Biodi-versidade”(SISBIO)forprovidingthecollectionlicense(processno. 29229).RFKissupportedbyCNPq(processno.308908/2016-3).

References

Allan,J.R.,Venter,O.,Maxwell,S.,Bertzky,B.,Jones,K.,Shi,Y.,Watson,J.E.M.,2017. RecentincreasesinhumanpressureandforestlossthreatenmanyNatural WorldHeritageSites.Biol.Conserv.206,47–55.

Azevedo,R.R.,Krüger,R.F.,2013.Theinﬂuenceoftemperatureandhumidityon abundanceandrichnessofCalliphoridae(Diptera).Iheringia.Ser.Zool.103, 145–152.

Azevedo,R.R.,Duarte,J.L.P.,Ribeiro,P.B.,Krüger,R.F.,2007.Occurrenceof Sarco-promuscapruna(Diptera)inSouthernBrazilasavectorofDermatobiahominis (Diptera)eggs.Arq.Bras.Med.Vet.Zootec.59,1348–1350.

(11)

Barnosky,A.D.,Hadly,E.A.,Bascompte,J.,Berlow,E.L.,Brown,J.H.,Fortelius,M.,Getz, W.M.,Harte,J.,Hastings,A.,Marquet,P.A.,Martinez,N.D.,Mooers,A., Roopnar-ine,P.,Vermeij,G.,Williams,J.W.,Gillepsie,R.,Kitzes,J.,Marshall,C.,Matzke,N., Mindell,D.P.,Revilla,E.,Smith,A.B.,2012.ApproachingastateshiftinEarth’s biosphere.Nature486,52–58.

Basset,Y.,Kitching,R.L.,1991.Speciesnumber,speciesabundanceandbodylengthof arborealarthropodsassociatedwithanAustralianrainforesttree.Ecol.Entomol. 16,391–402.

Bencke,G.A.,2009.Diversidadeeconservac¸ãodafaunadosCamposdoSuldoBrasil. In:Pillar,V.P.P.,Müller,S.C.,Castilhos,Z.M.S.,Jacques,A.V.A.(Eds.),Campos Sulinos:Conservac¸ãoeUsoSustentáveldaBiodiversidade.MinistériodoMeio Ambiente,Brasília,pp.101–121.

Bilenca,D.,Codesido,M.,González-Fischer,C.,Carusi,L.P.,Zuﬁaurre,E.,Abba,A., 2012.Impactosdelatransformaciónagropecuariasobrelabiodiversidadenla provinciadeBuenosAires.Rev.Mus.Argent.Cienc.Nat.14,189–198. Blondel,J.,2003.Guildorfunctionalgroups:doesitmatter?Oikos100,223–231. Borcard,D.,Gillet,F.,Legendre,P.,2011.NumericalEcologywithR.NewYork,

Springer-Verlag,http://dx.doi.org/10.1007/978-1-4419-7976-6.

Brown,B.V.,2005.Malaisetrapcatchesandthecrisisinneotropicaldipterology.Am. Entomol.513,180–183.

Bunge,J.,Fitzpatrick,M.,1993.Estimatingthenumberofspecies:areview.J.Am. Stat.Assoc.88,364–373.

Burger, M.I., 2000. Situação e ações prioritárias para a conservação de ban-hados e áreas úmidas da zona costeira. ANP, Brasília, Available at: https://www.brasil-rounds.gov.br/round7/arquivosr7/PERFURACAOR7/ refere/Banhados.pdf(accessed14.02.17).

Carvalho,C.J.B.,Couri,M.S.,2002.PartI.Basalgroups.In:Carvalho,C.J.B.(Ed.), Mus-cidae(Diptera)ofNeotropicalRegion:Taxonomy.EditoraUniversidadeFederal doParaná,Curitiba,pp.17–132.

Carvalho,C.J.B.,Almeida,J.R.,Jesus,C.B.,1984.DípterossinantrópicosdeCuritibae arredores(Paraná,Brasil).I.Muscidae.Rev.Bras.Entomol.28,551–560. Carvalho,C.J.B.,Bortolanza,M.,Silva,M.C.C.,Soares,E.D.G.,2003.Distributional

pat-ternsoftheNeotropicalMuscidae(Diptera).In:Morrone,J.J.,Bousquets,J.L. (Eds.),UnaperspectivaLatinoamericanadelaBiogeografía.,1ed.LasPrensas deCiencia,FacultaddeCienciasUniversidadAutónomadeMéxico,México,pp. 263–274.

Carvalho,C.J.B.,Couri,M.S.,Pont,A.C.,Pamplona,D.,Lopes,S.M.,2005.Acatalogue oftheMuscidae(Diptera)oftheNeotropicalRegion.Zootaxa860,1–282. Celulose Riograndense, 2012. Plano de Manejo da RPPNBarba Negra,

Avail-ableat:http://www.celuloseriograndense.com.br/responsabilidade/(accessed 26.06.15).

Chao,A.,1987.Estimatingthepopulationsizeforcapture–recapturedatawith unequalcatchability.Biometrics43,783–791.

Chao,A.,Colwell,R.K.,Lin,C.W.,Gotelli,N.J.,2009.Sufﬁcientsamplingforasymptotic minimumspeciesrichnessestimators.Ecology90,1125–1133.

Chiu,C.H.,Wang,Y.T.,Walther,B.A.,Chao,A.,2014.Animprovednonparametric lowerboundofspeciesrichnessviaamodiﬁedgood-turingfrequencyformula. Biometrics70,671–682.

Coelho,S.M.P.,1997.AspectosmorfológicosdaProbóscidedeMuscoidea(Diptera, Muscomorpha).Rev.Bras.Zool.14,245–253.

Colwell,R.K.,Coddington,J.A.,1994.Estimatingterrestrialbiodiversitythrough extrapolation.Philos.Trans.R.Soc.LondonSer.B345,101–118.

Colwell,R.K.,Mao,C.X.,Chang,J.,2004.Interpolating,extrapolating,andcomparing incidence-basedspeciesaccumulationcurves.Ecology85,2717–2727. Costacurta,N.C., Carvalho,C.J.B.,2005. TaxonomyofNeurotrixa ShannoneDel

Ponte(Diptera:Muscidae)withdescriptionofnewspeciesfromsouthernBrazil. Neotrop.Entomol.34,927–932.

Costacurta,N.C.,Couri,M.S.,Carvalho,C.J.B.,2005.Descriptionsofnewspecieswith akeytoidentiﬁcationofthegenusNeodexiopsisMalloch(Diptera,Muscidae)in Brazil.Rev.Bras.Entomol.49,353–366.

Costacurta,N.C.,Marinoni,R.C.,Carvalho,C.J.B.,2003.FaunadeMuscidae(Diptera) emtrês localidades doEstado doParaná,Brasil,capturadaporarmadilha Malaise.Rev.Bras.Entomol.47,389–397.

Couri,M.S.,Carvalho,C.J.B.,2002.PartII.Apicalgroups.In:Carvalho,C.J.B.(Ed.), Mus-cidae(Diptera)ofNeotropicalRegion:Taxonomy.EditoraUniversidadeFederal doParaná,Curitiba,pp.133–286.

Crawshaw,D.,Dall’agnol,M.,Cordeiro,J.L.P.,Hasenack,A.,2007.Caracterizacão doscampossul-rio-grandenses:umaperspectivadaecologiadapaisagem.Bol. GaúchoGeogr.33,233–252.

DaSilva,P.G.,Vaz-De-Mello,F.Z.,DiMare,R.A.,2013.Diversityandseasonalityof Scarabaeinae(Coleoptera:Scarabaeidae)inforestfragmentsinSantaMaria,Rio GrandedoSul,Brazil.An.Acad.Bras.Cienc.85,679–697.

Duarte,J.L.P.,Krüger,R.F.,Carvalho,C.J.B.,Ribeiro,P.B.,2010.Evidenceofthe inﬂu-enceofMalaisetrapageonitsefﬁciencyinthecollectionofMuscidae(Insecta, Diptera).Int.J.Top.Insect.Sci.30,115–118.

Erwin,T.L.,1998.Thetropicalforestcanopy.Theheartofbioticdiversity.In:Wilson, E.O.(Ed.),Biodiversity.NationalAcademyPress,Washington,pp.109–127. Ferrar,P.,1987.AGuidetotheBreedingHabitsandImmatureStagesofDiptera

Cyclorrhapha.ScandinavianSciencePress,Copenhagen.

Ferreira,J.,Pardini,R.,Metzger,J.P.,Fonseca,C.R.,Pompeu,O.S., Sparovek,G., Louzada,J.,2012.TowardsenvironmentallysustainableagricultureinBrazil: challengesandopportunitiesforappliedecologicalresearch.J.Appl.Ecol.49, 535–541.

Filloy,J.,Bellocq,M.I.,2006.SpatialvariationsintheabundanceofSporophiia seedeatersinthesouthernNeotropics:contrastingtheeffectsofagricultural developmentandgeographicalposition.Biodivers.Conserv.15,3329–3340.

Foley,J.A.,DeFries,R.,Asner,G.P.,Barford,C.,Bonan,G.,Carpenter,S.R.,Chapin,F.S., Coe,M.T.,Daily,G.C.,Gibbs,H.K.,Helkowski,J.H.,Holloway,T.,Howard,E.A., Kucharik,C.J.,Monfreda,C.,Patz,J.A.,Prentice,I.C.,Ramankutty,N.,Snyder,P.K., 2005.Globalconsequencesoflanduse.Science309,570–574.

Fontana,C.S.,Bencke,G.A.,Reis,R.E.,2003.Livrovermelhodafaunaameac¸adade extinc¸ãonoRioGrandedoSul.EDIPUCRS,PortoAlegre.

Gonzalez, S., Merino, M.L., 2008. Ozotoceros bezoarticus. In: IUCN 2013. IUCN Red List of Threatened Species. Version 2013.2, Available at: http://www.iucnredlist.org(accessed18.03.14).

Gotelli,N.J.,Colwell,R.K.,2001.Quantifyingbiodiversity:proceduresandpitfallsin themeasurementandcomparisonofspeciesrichness.Ecol.Lett.4,379–391. Greenberg,B.,1971.FliesandDisease.Ecology,ClassiﬁcationandBioticAssociations.

PrincetonUniversityPress,NewJersey.

Hochmüller, C.J., Lopes-Da-Sila, M., Valente, V.L.S., Schmitz, H.J., 2010. The drosophilidfauna(Diptera,Drosophilidae)ofthetransitionbetweenthePampa andAtlanticForestBiomesinthestateofRioGrandedoSul,southernBrazil: ﬁrstrecords.Pap.AvulsosZool.50,285–295.

Hsieh,T.C.,Ma,K.H.,Chao,A., 2016.iNEXT:anRpackageforrarefactionand extrapolation of species diversity (Hill numbers). Methods Ecol. Evol. 7, 1451–1456.

IBGE–InstitutoBrasileirodeGeografiaeEstatística,1986.Vegetação,UsoPotencial daTerra.In:IBGE,LevantamentodeRecursosNaturais,RiodeJaneiro,pp.1–796. IBGE – Instituto Brasileiro de Geografia e Estatística, 2004. Mapa de vegetação do Brasil, Brasília, Available at: https://ww2.ibge.gov.br/ home/presidencia/noticias/21052004biomas.shtm(accessed30.01.18). ICMBIO – Instituto Chico Mendes de Conservação da Biodiversidade, 1999.

Plano de Manejo do Parque Nacional da Lagoa do Peixe, Available at: http://www.icmbio.gov.br/portal/images/stories/imgs-unidades-coservacao/ parnalagoa-do-peixe.pdf(accessed02.08.16).

Kindt,R.,Van-Damme,P.,Simons,A.J.,2006.TreediversityinwesternKenya: usingproﬁlestocharacterizerichnessandevenness.Biodivers.Conserv.15, 1253–1270.

Kirst,F.D.,Marinoni,L.,Krüger,R.F.,2015.Newdistributionrecordsfor Sciomyzi-daespecies(Insecta,Diptera)fromRioGrandedoSul,Brazil.CheckList11, 1–5.

Kirst,F.D.,Marinoni,L.,Krüger,R.F.,2017.WhatdoestheSouthernBrazilianCoastal Plaintellaboutitsdiversity?Syrphidae(Diptera)asamodel.Bull.Entomol.Res. 107,645–657.

Krapovickas, S., Di Giacomo, A.S., 1998. Conservation of pampas and campos grasslands in Argentina. Parks 8, 47–53, Available at: https://www.iucn.org/sites/dev/ﬁles/import/downloads/parksoct981.pdf (accessed30.01.18).

Krüger,R.F.,Krolow,T.K.,2015.Seasonalpatternsofhorseﬂyrichnessand abun-danceinthePampabiomeofsouthernBrazil.J.VectorEcol.40,364–372. Krüger,R.F.,Carvalho,C.J.B., Ribeiro, P.B.,2010.Assembly rulesinmuscid ﬂy

assemblagesinthegrasslandsbiomeofSouthernBrazil.Neotrop.Entomol.39, 345–353.

Leite,P.F.,2002.Contribuic¸ãoaoconhecimentoﬁtoecológicodosuldoBrasil.Ciência andAmbiente24,52–73.

Löwenberg-Neto,P.,Carvalho,C.J.B.,2009.Areasofendemismandspatial diversiﬁ-cationoftheMuscidae(Insecta:Diptera)intheAndeanandNeotropicalregions. J.Biogeogr.36,1750–1759.

Löwenberg-Neto,P.,Carvalho,C.J.B.,2013.Muscidae(Insecta:Diptera)ofLatin AmericaandtheCaribbean:geographicdistributionandcheck-listbycountry. Zootaxa3650,1–147.

Machado,A.B.M.,Drummond,G.M.,Paglia,A.P.,2008.LivroVermelhodaFauna BrasileiraAmeac¸adadeExtinc¸ão.Biodiversidade19.

Magurran,A.E.,2004.MeasuringBiologicalDiversity.OxfordBlackwellScience, Oxford.

Marques,B.,Couri,M.S.,2007.Taxonomiaemorfologiadeespéciesneotropicais deGraphomyaRobineau-Desvoidy(Diptera,Muscidae).Rev.Bras.Entomol.51, 436–444.

McAlpine,J.F.,1981.Morphologyandterminology:adults.In:McAlpine,J.F., Peter-son,B.V.,Shewell,G.E.,Teskey,H.J.,Vockeroth,J.R.,Wood,D.M.(Eds.),Manual ofNearticDiptera.Ottawa,pp.9–63.

McLintock,J.,Depner,K.R.,1954.Areviewofthelife-historyandhabitsofthehorn ﬂy,Siphonairritans(L.)(Diptera:Muscidae).Can.Entomol.86,20–33. Meira,J.R.,Porto,M.L.,1998.ReservaBiológicadoLami:avidanabeiradolago.In:

Menegat,R.(Ed.),AtlasAmbientaldePortoAlegre.EditoraUFRGS,PortoAlegre, pp.89–92.

Mello-Filho,L.E.,Somner,G.V.,Peixoto,A.L.,1992.CenturiaPlantarumBrasiliensium ExstintionisMinitata.SociedadeBotânicadoBrasil/Ibama,Brasília.

Mikich,S.B.,Bérnils,R.S.,2004.LivroVermelhodaFaunaAmeac¸adanoEstadodo Paraná,Availableat:http://www.meioambiente.pr.gov.br/arquivos/File/cobf/ livrofaunaextincao.pdf(accessed14.02.17).

MMA–MinistériodoMeioAmbiente,2000.Avaliaçãoeaçõesprioritáriaspara aconservaçãodabiodiversidadedaMataAtlânticaeCamposSulinos, Avail-ableat:http://www.mma.gov.br/estruturas/sbfchmrbbio/arquivos/Sumario %20Mata%20Atlantica.pdf(accessed17.02.17).

MMA – Ministério do Meio Ambiente, 2004. Federal Decree n◦ _5.092, _May

21, 2004 , Available at: http://www2.camara.leg.br/legin/fed/decret/2004/ decreto-5092-21-maio-2004-532394-publicacaooriginal-14437-pe.html (accessed14.02.17).

Nihei,S.S.,2005.AreviewoftheNeotropicalgenusSarcopromuscaTownsend (Diptera:Muscidae)withakeytospeciesandaredescriptionofS.sarcophagina (Wulp).Zootaxa1004,51–64.

(12)

Nihei,S.S.,Carvalho,C.J.B.,2005.DistributionalpatternsoftheNeotropicalﬂygenus PolietinaSchnablandDziedzicki(Diptera,Muscidae):aphylogeny-supported analysisusingpanbiogeographictools.Pap.AvulsosZool.45,313–326. Nihei,S.S.,Carvalho,C.J.B.,2007.SystematicsandbiogeographyofPolietinaSchnabl

andDziedzicki(Diptera,Muscidae):Neotropicalarearelationshipsand Amazo-niaasacompositearea.Syst.Entomol.32,477–501.

Novotn ´y,V.,1993.Spatialandtemporalcomponentsofspeciesdiversityin Auchen-orrhyncha(Insecta:Hemiptera)communitiesofIndochinese montanerain forest.J.Trop.Ecol.9,93–100.

Oksanen,J., Blanchet,F.G., Kindt, R., Legendre, P.,Minchin, P.R., O’Hara,R.B., Simpson, G.L., Solymos, P., Henry, M., Stevens, H., Wagner, H., 2015. vegan:CommunityEcologyPackage.RPackageVersion2.3-0,Availableat: https://github.com/vegandevs/vegan(accessed07.08.18).

Overbeck,G.E.,Muller,S.C.,Fidelis,A.,Pfadenhauer,J.,Pillar,V.D.,Blanco,C.C., Boldrini,I.I.,Both,R.,Forneck,E.D.,2007.Brazil’sneglectedbiome:theSouth BrazilianCampos.PlantEcol.Evol.Syst.9,101–116.

Parmar,T.K.,Rawtani,D.,Agrawal,Y.K.,2016.Bioindicators:thenaturalindicatorof environmentalpollution.FrontLifeSci.9,110–118.

Paz,A.L.G.,Romanowski,H.P.,deMorais,A.B.B.,2013.DistributionofSatyrini (Lepi-doptera,Nymphalidae)inRioGrandedoSulState,southernBrazil.Ecol.Res.28, 417–426.

Pedroso-De-Paiva,D.,1996.AspectosdabiologiadeadultosdeSarcopromuscapruna (ShannonandDelPonte,1926)(Diptera:Muscidae).Rev.Bras.Biol.56,183–190. Pereira, H.M., Leadley, P.W., Proenc¸a, V., Alkemade, R., Scharlemann, J.P.W., Fernandez-Manjarrés,J.F.,Araújo,M.B.,Balvanera,P.,Biggs,R.,Cheung,W.W.L., Cini,L.,Cooper,H.D.,Gilman,E.L.,Guénette,S.,Hurtt,G.C.,Huntington,H.P., Mace,G.M.,Oberdorff,T.,Revenga,C.,Rodrigues,P.,Scholes,R.J.,Sumaila,U.R., Walpole,M.,2010.ScenariosforGlobalBiodiversityinthe21stCenturyScience. Science330,1496–1501.

Pereira-Colavite,A.,Carvalho,C.J.B.,2012.TaxonomyofNeomuscinaTownsend (Diptera,Muscidae)fromBrazil.Zootaxa3504,1–55.

Pillar,V.P.P.,Müller,S.C.,Castilhos,Z.M.S.,Jacques,A.V.A.,2009.CamposSulinos: Conservac¸ãoeUsoSustentáveldaBiodiversidade.MinistériodoMeioAmbiente, Brasília.

Poppe,J.L.,Valente,V.L.S.,Schmitz,H.J.,2012.StructureofDrosophilidaeassemblage (Insecta,Diptera)inPampaBiome(SãoLuizGonzaga,RS).Pap.AvulsosZool.52, 185–195.

Poppe,J.L.,Valente,V.L.S.,Schmitz,H.J.,2013.Populationdynamicsofdrosophilids inthepampabiomeinresponsetotemperature.Neotrop.Entomol.42,269–277. RDevelopmentCoreTeam,2015.R:ALanguageandEnvironmentforStatistical

Computing,Avaiableat:http://www.R-project.org(accessed26.11.13). Rodríguez-Fernández,J.I.,Carvalho,C.J.B.,Moura,M.O.,2006.Estruturade

assem-bléiasdeMuscidae(Diptera)noParaná:umaanálisepormodelosnulos.Rev. Bras.Entomol.50,93–100.

Root,R.,1967.Thenicheexplorationpatternoftheblue-graygnatcatcher.Ecol. Monogr.37,317–350.

Rosado,J.L.O.,Gonc¸alves,M.G.,Dröse,W.,Silva,E.J.E.,Krüger,R.F.,Feitosa,R.M., Loeck,A.E.,2012.Epigeicants(Hymenoptera:Formicidae)invineyardsand grasslandareasintheCampanharegion,stateofRioGrandedoSul,Brazil.Check List8,1184–1189.

Rosado,J.L.O.,Gonc¸alves,M.G.,Dröse,W.,Silva,E.J.E.,Krüger,R.F.,Feitosa,R.M., Loeck,A.E.,2013.Effectofclimaticvariablesandvinecropsontheepigeicant fauna(Hymenoptera:Formicidae)intheCampanharegion,stateofRioGrande doSul,Brazil.J.InsectConserv.17,1113–1123.

Santos,A.M.C., Whittaker,R.J.,Triantis,K.A.,Jones,O.R.,Borges,P.A.V.,Quicke, D.L.J.,Hortal,J.,2010.Arespecies–arearelationshipsfromentirearchipelagos congruentwiththoseoftheirconstituentislands?GlobalEcol.Biogeogr.19, 527–540.

Schüehli,G.S.,Carvalho,C.J.B., 2005.RevisionandcladisticsoftheNeotropical genus PseudoptilolepisSnyder(Diptera, Muscidae).Rev.Bras. Entomol. 22, 23–34.

SEMA–SecretariaEstadualdoMeioAmbiente,1996.PlanodeManejodo Par-queEstadualItapuã,Availableat:http://www.sema.rs.gov.br/upload/arquivos/ 201610/15171153-plano-manejo-peitapua.pdf(accessed14.02.17).

SEMA–SecretariaEstadualdoMeioAmbiente,2013.ParquesEstaduais,Availableat: http://www.sema.rs.gov.br/conteudo.asp?codmenu=174(accessed02.08.15). Skidmore,P.,1985.TheBiologyofMuscidaeoftheWorld.JunkPublishers,Dordrecht. Sneath,P.H.,Sokal,R.R.,1973.NumericalTaxonomy:ThePrinciplesandPracticeof

NumericalClassiﬁcation.W.H.FreemanandCompany,SanFrancisco. Townes,H.A.,1972.Alight-weightMalaisetrap.Entomol.News83,239–247. Ugland,K.I.,Gray,J.S.,Ellingsen,K.E.,2003.Thespeciesaccumulationcurveand

estimationofspeciesrichness.J.Anim.Ecol.72,888–897.

Venzke,T.S.,2012.FlorísticadecomunidadesarbóreasnoMunicípiodePelotas,Rio GrandedoSul.Rodriguésia63,571–578.

Waechter,J.L.,1985.Aspectosecológicosdavegetac¸ãodeRestinganoRioGrande doSul,Brasil.Comun.Mus.Cienc.Tecnol.Pucrs.Ser.Bot.33,49–68.

Waechter,J.L.,2002.PadrõesgeográﬁcosnaﬂoraatualdoRioGrandedoSul.Ciência andAmbiente24,93–108.

Weisse,M.J.,Naughton-Treves,L.C.,2016.Conservationbeyondparkboundaries:the impactofbufferzonesondeforestationandminingconcessionsinthePeruvian Amazon.Environ.Manage.58,297–311.

Werner, D., Pont, A.C.,2006. Thefeeding and reproductive behaviour ofthe Limnophorinii(Diptera:Muscidae).In:ProceedingsoftheInternational Sym-posiumonSimuliidae.Supplement.Stud.Dipterol.14,pp.79–114.

Willie,J.,Petre,C.A.,Tagg,N.,Lens,L.,2012.Evaluationofspeciesrichnessestimators basedonquantitativeperformancemeasuresandsensitivitytopatchinessand samplegrainsize.ActaOecol.45,31–41.

Witt,P.B.R.,2013.FaunaeFloradaReservaBiológicaLamiJoséLützenberger. Sec-retariaMunicipaldoMeioAmbiente,PortoAlegre.

Zafalon-Silva,Â.,Hoffmeister,C.H.,Anjos,V.A.,Ribeiro,P.B.,Krüger,R.F.,2014. NecrophagousDipteraassociatedwithwildanimalcarcassesinsouthernBrazil. Rev.Bras.Entomol.58,337–342.