research article broad protein spectrum in stored pollen...

Research ArticleBroad Protein Spectrum in Stored Pollen ofThree Stingless Bees from the Chaco Dry Forest inSouth America (Hymenoptera Apidae Meliponini) andIts Ecological Implications

Favio Gerardo Vossler

Laboratorio de Actuopalinologıa CICyTTP-CONICETFCyT-UADER Dr Materi y Espana DiamanteE3105BWA Entre Rıos Argentina

Correspondence should be addressed to Favio Gerardo Vossler favossleryahoocomar

Received 26 September 2015 Accepted 2 November 2015

Academic Editor David Roubik

Copyright copy 2015 Favio Gerardo Vossler This is an open access article distributed under the Creative Commons AttributionLicense which permits unrestricted use distribution and reproduction in any medium provided the original work is properlycited

Protein content of pollen stored by three meliponine species was variable from 978 (less than half the value considered as optimalto brood development in Apis mellifera) in type Capparis tweediana-C speciosa to more than 26 in typeMaytenus vitis-idaea andsome Prosopis samplesThis pollen of low protein value was occasionally foraged (only six out of 75masses analyzed ofG argentinabut none in 86 masses of T fiebrigi or in ten ofM orbignyi) However it is likely that amino acid deficiencies of certain pollens arecompensated by randomly foraging on a broad spectrum of pollen plants The large amounts of pollen stored in their nests mightalso be important in compensating these deficiencies The only sample studied forM orbignyi showed a protein value greater thanthe one required for A mellifera and was dominated by types Acacia praecox and Prosopis As this species also prefers Solanumand other protein-rich pollen more samples would need to be analyzed to establish whether protein requirements are high for thisMelipona species Pollen showing the highest protein content (gt26) belonged to highly nectariferous plants well represented inmeliponine and Apis honey such as ProsopisMaytenus and Ziziphus

1 Introduction

The stingless bees (Apidae Meliponini) are eusocial andthey build perennial nests with hundreds to thousands ofindividuals and high reproductive rate [1 2] To main-tain their colonies high amounts of pollen and nectar areforaged from flowers the former stored as pollen massesand the latter as honey in pots made of cerumen Pollenpots are located mainly surrounding the brood area whereyoung individuals are growing [3] Pollen stored in nestsis chemically and biochemically different from fresh pollenfrom anthers or pollen loads from the same plant speciesdue to regurgitated liquids incorporated during packing [4ndash7] Mutualistic bacteria were found in stored pollen andhoney of nine Apis species Melipona fasciata panamicaCockerell Melipona beecheii Bennett Meliponula bocandei(Spinola) and Trigona producing enzymes that may facilitate

storage andor digestion by bees and secrete antibiotics andfatty acids to inhibit microorganisms that cause spoilage ofstored food [8 9] During periods of general food shortageMelipona species show conservative trends eating younglarvae and provisions and drastically decreasing brood pro-duction [10] Although exceptional protein resources suchas soybean bran in Geotrigona mombuca (Smith) (sub Ginusitata in [11]) and flesh of dead animals in many TrigonaPartamona Oxytrigona Cephalotrigona and others [12] canbe facultatively gathered pollen is the main protein sourcein bees Obligate necrophagy (dead animal flesh is the onlyprotein source) occurs only in Trigona crassipes (Fabricius)T hypogea Silvestri and T necrophaga Camargo amp Roubik[11 13ndash15] For Apis mellifera the higher the crude proteinpercentage the lesser amount of pollen required to sustainproduction [16] Ample protein content promotes a high birthrate and long-lived bees 20 being the minimum protein

Hindawi Publishing CorporationPsycheVolume 2015 Article ID 659538 7 pageshttpdxdoiorg1011552015659538

2 Psyche

amount required by colonies for optimum production [17]A well-nourished bee colony is a guarantee for good level ofhoney flow and breeding [16]

It is important to know pollen protein intake of melipo-nine bees for appropriate colony rearing (meliponiculture)as it would be useful to determine supplemental feedingneeded in periods of flowering shortage Stingless bees areimportant for pollination of wild and crop plants [18] and forbee-products production (honey pollen resin and cerumen)[19] The aims of the present study were to determine proteincontent of pollen stored in nests of three meliponine beespecies (Tetragonisca fiebrigi (Schwarz) Melipona orbignyi(Guerin) andGeotrigona argentinaCamargo ampMoure) fromthe South American Chaco dry forest These are three outof the seven species recorded for this area [20 21] beingimportant mainly for rural population for their honey pollenmasses larvae and cerumen [22]

2 Materials and Methods

21 Sampling and Study Area Pollen mass from ceru-men pots was obtained from nests of Tetragonisca fieb-rigi (Schwarz) Melipona orbignyi (Guerin) and Geotrigonaargentina Camargo amp Moure from xeric forests in the Chacoregion of Northern Argentina (Figure 1) Samples for proteinanalysis were taken from nest 7 of T fiebrigi nest 11 ofM orbignyi and nests 2 and 4 of G argentina from ElSauzalito (24∘241015840S 61∘401015840W) from nest 1 of G argentinafrom El Espinillo (25∘ 241015840S 60∘ 27W) from nest 5 ofT fiebrigi from Miraflores (25∘291015840S 61∘011015840W) and fromnest 12 of T fiebrigi from Villa Rıo Bermejito (25∘371015840S60∘151015840W) This Chaco dry forest is characterized by thedominance of ldquopalo santordquo (Bulnesia sarmientoi Lorentz exGriseb Zygophyllaceae) and ldquoquebracho blancordquo trees (Aspi-dosperma quebracho-blanco Schltdl Apocynaceae) or byldquoquebracho colorado santiaguenordquo (Schinopsis lorentzii EnglAnacardiaceae) ldquoquebracho colorado chaquenordquo (Schinopsisbalansae Engl) and ldquoquebracho blancordquo [23 24] Otherwoody elements well represented in this xerophilous flora areldquomistolrdquo (Ziziphus mistol Griseb Rhamnaceae) ldquomollerdquo orldquoguaraninardquo (Sideroxylon obtusifolium (Roem amp Schult) TD Penn Sapotaceae) several species of Prosopis (ldquoalgarroboblancordquo ldquoalgarrobo negrordquo ldquovinalrdquo ldquovinalillordquo ldquocarandardquoand ldquoguachınrdquo) (Fabaceae Mimosoideae) ldquoguayacanrdquo (Cae-salpinia paraguariensis (D Parodi) Burkart Fabaceae Cae-salpinioideae) ldquotipa coloradardquo or ldquopalo cocardquo (Pterog-yne nitens Tul Fabaceae Caesalpinioideae) ldquopalo cruzrdquo(Tabebuia nodosa (Griseb) Griseb Bignoniaceae) ldquoque-brachillordquo (Aspidosperma triternatum Rojas Acosta Apocy-naceae) ldquopalo borrachordquo or ldquoyuchanrdquo (Ceiba chodatii (Hassl)Ravenna Bombacaceae) ldquopalma de monterdquo (Trithrinaxschizophylla Drude Arecaceae) ldquomeloncillordquo (Castela coc-cinea Griseb Simaroubaceae) ldquopalo tintardquo (Achatocarpuspraecox Griseb Achatocarpaceae) ldquotalardquo (Celtis spp Celti-daceae) ldquoduraznillordquo (Salta triflora (Griseb) Adr SanchezPolygonaceae) ldquopatardquo (Ximenia americana L Olacaceae)ldquomollerdquo (Schinus fasciculatus (Griseb) I M Johnst vararenicola (Hauman) F A Barkley Anacardiaceae) ldquochanarrdquo

(Geoffroea decorticans (Gillies ex Hook amp Arn) BurkartFabaceae Papilionoideae) ldquosal de indiordquo or ldquosal de monterdquo(Maytenus vitis-idaea Griseb Celastraceae) ldquopaloma yuyordquo(Moya spinosa Griseb Celastraceae) ldquoteatınrdquo (Mimosa deti-nens Benth Fabaceae Mimosoideae) ldquocardonrdquo (Stetsoniacoryne (Salm-Dyck) Britton amp Rose Cactaceae) ldquouclerdquo(Cereus forbesii Otto ex C F Forst Cactaceae) twospecies of Bougainvillaea (ldquorama overardquo) (Nyctaginaceae)several species of Acacia (ldquotuscardquo ldquogarabatordquo and ldquochurquirdquo)(Fabaceae Mimosoideae) Capparis (ldquoatamisquirdquo ldquosachamembrillordquo ldquosacha sandiardquo ldquosacha porotordquo and ldquobolaverderdquo) and ldquocardordquo or ldquochaguarrdquo (several Bromeliaceaegenera) among others

22 Pollen Analysis of Pollen Masses and Plant and BeeReferences Pollen masses were dissolved in distilled water at80ndash90∘C and stirred with a magnetic stirrer for 10ndash15minA representative mixture of 5ndash10mL was obtained and cen-trifuged at 472timesg for 5min Processing included acetolysis[25] Under a Nikon Eclipse E200 light microscope a total of300ndash500 pollen grains per slide were counted Pollen grainidentification was carried out comparing nest pollen slideswith those present in the reference pollen collection Thereference slides consist of a total of 190 plant species and aredeposited in PAL-CICYTTP pollen collection of DiamanteEntre Rıos Argentina It was made from flower buds of plantspecies collected in various localities from Chaco Province ofArgentina These plant specimens were pressed dried andidentified by the author and deposited in the Herbariumof the Museo de La Plata (LP) the Herbarium of MuseoArgentino de Ciencias Naturales ldquoBernardino Rivadaviardquo(BA) Buenos Aires and the Herbarium Lorentz (DTE) ofDiamante Entre Rıos Argentina Plant nomenclature follows[26] Bee specimens were collected from nests identifiedby Arturo Roig-Alsina and deposited in the EntomologyCollection of the Museo Argentino de Ciencias NaturalesldquoBernardino Rivadaviardquo Buenos Aires Argentina

23 Determination of Protein Content To quantify proteinof pollen resources foraged by bees certain pollen masseswere selected according to their pollen type compositionSix pollen mass samples were taken from nests 5 7 and12 of Tetragonisca fiebrigi one from nest 11 of Meliponaorbignyi and eight from nests 1 2 and 4 of Geotrigonaargentina (Table 1) For nitrogen content determination50mg of pollen [27] was analyzed by the micro-Kjeldahlmethod [28] and crude protein was estimated using thefactor 625 [29] Pollen nitrogen content was analyzed inLANAIS N15 (National Laboratory of Research and ServicesUNS-CONICET) Departamento de Agronomıa Universi-dad Nacional del Sur Bahıa Blanca Argentina

3 Results

31 Protein Content of Pollen Stored in Meliponine NestsNitrogen values from pollen grains stored in pollen potsof the three meliponine species studied varied from 156to 486 which is equivalent to 978 to 3041 of crude

Psyche 3

Table1Mainpo

llentypes(gt10representatio

n)presentinpo

llenmasssam

ples

ofTetra

goniscafiebrigi(Tf

)Melipona

orbignyi(M

o)and

Geotrigona

argentina(G

a)andtheirn

itrogen

and

proteinpercentages

Bees

pecies

Sample

Nestand

pollenmass

Mainpo

llentypes

Nitrogen

()

Crud

eprotein

()

Tf

15B

Type

Schinopsis(59

)+Trith

rinax

schizophylla(405

)3055

1909

25C

Trith

rinax

schizophylla(41

)+Pa

rthenium

hyste

rophorus

(30

)+type

Schinopsis(28

)2854

1784

35E

Trith

rinax

schizophylla(98

)2898

1811

47A

Sideroxylonobtusifolium

(24

)+Prosopis(18

)+type

Maytenu

svitis-idaea

(16

)+Tabebuia(13

)3126

1954

57F

Capparisspeciosa

(42

)+Mascagniabrevifolia

(34

)+Heliantheae

(10

)364

82280

612

CZizip

husm

istol(60

)+Prosopis(30

)3542

2214

Mo

711

Type

Acaciapraecox(53

)+Prosopis(38

)3881

2426

Ga

8458

Type

Capparistweedian

a-C

speciosa

(100)

156

978

912

4Ca

stelacoccinea

(100)

326

2036

10231

Prosopis(100)

334

2087

11111

Prosopis(91

)486

3041

12236

Prosopis(895

)

432

2703

13233

Type

Maytenu

svitis-idaea

(100)

423

2687

14242

Type

Maytenu

svitis-idaea

(96

)424

2651

15116

Type

Croton

(30

)+type

Sagitta

ria(18

)+Eleocharis(12

)+Ca

stelacoccinea

(11

)265

1659

4 Psyche

(a) (b) (c)

(d) (e) (f)

Figure 1 General aspect of pollen masses of Tetragonisca fiebrigi (a b) Melipona orbignyi (c) and Geotrigona argentina (d e) Individualloads from pollen masses can be seen in scanning microscope (f) Bars (a) 10mm (b) 5mm (c) 20mm (d) 10mm (e) 20mm (f) 1mm

protein (Table 1) Higher protein values (gt20)were found insamples having the following composition Prosopis (samples10 11 and 12 of G argentina) type Maytenus vitis-idaea(samples 13 and 14 of G argentina) type Acacia praecox+ Prosopis (sample 7 of M orbignyi) Capparis speciosa +Mascagnia brevifolia + Heliantheae (sample 5 of T fiebrigi)Ziziphus mistol + Prosopis (sample 6 of T fiebrigi) andCastela coccinea (sample 9 of G argentina) (Table 1) Theremaining samples showed less than 20of crude protein thelowest value (lt10) found was for type Capparis tweediana-C speciosa (sample 8 of G argentina)

Protein values found in samples composed mainly ofherbs climbers or shrubs differed greatly among them (978ndash228) and were slightly lower than those dominated by trees(1784ndash3041) A broader range of protein values was foundfor G argentina (978ndash3041) than for T fiebrigi (1784ndash2280)

4 Discussion

If protein content of pollen grainswas constant for all individ-uals of each plant species samples composed only or mostlyof one pollen type (96ndash100) (6 samples in the present study)would indicate their real protein value and it would be usefulfor comparing samples composed of many pollen types

Nutritive value of pollen grains is affected by air temperaturesoil moisture pH and soil fertility among other factors [30]but a range of values is expected for a particular species underthe same conditions [16] The two samples dominated bytype Maytenus vitis-idaea had similarly high protein values(2651 and 2687) followed by Castela coccinea that rankednear 20 the minimum limit value considered as optimalto brood development in Apis mellifera L [17] and the palmtree Trithrinax schizophylla with slightly less than 20 Thelowest protein value was from type Capparis tweediana-Cspeciosa with less than half the optimal value Inconsistencyamong pollen type composition and protein value was foundfor the three samples dominated by Prosopis as pure Prosopispollen showed lower protein values than those composedof 91 and 895 Prosopis and approximately 10 of othertypes (92 Ziziphus mistol in sample 11 and 95 Capparissalicifolia+ 1Ruprechtia triflora in sample 12) It appears thatthese accompanying pollen typeswere providers of importantamounts of proteins Nevertheless the differences of proteincontent detected in these three samples could be due tothe fact that different species of Prosopis (similar at lightmicroscope) were present in their composition as manyProsopis species are highly abundant in arboreal and shrubbystrata of the Chaco forest andmeliponine bees forage on all ofthem On the other hand soil fertility can influence nitrogen

Psyche 5

and consequent protein composition of plant individuals[30]The dry Chaco is a xerophytic forest that alternates withwater bodies such as ancient rivers and their related riparianvegetation [31] Nests ofmeliponine species here studied weresampled from these environments and bees could forageon flowers from both dry forest patches and water bodiescomposed of plants growing under different nitrogen levelMoreover livestock grazing in the forest is a very commonpractice in the area studied and it is responsible for increasingsoil nitrogen levels through excreta Local differences ofnitrogen levels in soil might be the cause for differences inprotein amounts found in pollenmasses dominated by a samepollen type (the case of Prosopis) Fertilizer incidence wasdiscarded as agricultural crops were absent in the sampledarea

Stored pollen in Apis mellifera colonies (pollen bread)is probably fermented by lactic acid bacteria of the generaLactobacillus and Bifidobacterium from the honey stomachadded to the pollen via regurgitated nectar [6 7] Overand above their significance in pollen bread productionand storage these lactic acid bacteria are important againstpathogens and production and storage of honey [6 9] Thesemicrobial symbionts are also present in its ecologically similarand closely related group the stingless bees (Meliponini) [932] Proteins mainly enzymes are secreted by this beneficialmicrobiota during stress [33] which could increase the pro-tein content of stored pollen Versatile digestive physiologiescharacterize broad polylectic bees [34] as pollen of diverseprotein spectrum seems to be similarly foraged by thesehighly social species Similar to other studies on bee foragedpollen [35 36] protein content of pollen types was variableIt is likely that amino acids deficiencies of certain pollen arecompensated by randomly foraging on a broad spectrum ofpollen plants This is in agreement with the argument statedby [37] that nutritive value of pollen for bees is not directlycorrelated with protein quantity since a qualitative factor isof greater importance For instance [38] detected low proteinlevels in hand-collected pollen from desert plants (from 7 to156) but they highlighted that their amino acid patternswere in agreement with the requirements for honey beesAn interesting topic to be tested by experimental studies ison the capability of meliponine brood to develop by eatingpollen having the half of the optimal protein level for Apismellifera However palynological surveys on meliponinebees from the Chaco region showed that scarce number ofpollen masses are composed of pure or nearly pure pollenfrom Capparis tweediana or C speciosa having only 978of protein (only six out of 75 masses analyzed ofG argentinabut none in 86 masses of T fiebrigi or in ten of M orbignyi)[39 FGV unpublished data] Pollen with low protein levelswould expose bees to more severe amino acids deficienciesHowever bees can be well developed when feeding on highamounts of these pollens but a colony would be threatenedwhen there is a low amount of pollen stores or shortageof flowerings [16] Meliponine bees store great amounts ofpollen (353 g in 20 pollen pots of diverse filling in one nestof G argentina) [39] (Figure 1)

An attempt was made to determine the differences inprotein content due to life-form of plants foraged and season

when sampling was carried out and among bee species butfurther studies are necessary However most protein-richpollen species were woody in accordance with findings forother semiarid areas of Argentina [35] but findings by [36]did not show this pattern It is widely known that Solanumspecies are protein-rich pollen resources ranging from 341for S tabanoense Correll to 549 for S lycopersicum L[40] Solanumwas highly foraged byMelipona orbignyi [FGVunpublished data] The only sample studied for M orbignyishowed a protein value greater than the one required for Amellifera and was dominated by types Acacia praecox andProsopis followed by four types of less than 4 representationAs this species also prefers Solanum and other protein-rich pollen from poricidal anthers whose grains are easilygathered by vibratile buzzing more samples would need tobe analyzed to establish whether protein requirements arehigh for this Melipona species The genus Solanum is wellrepresented in the dry Chaco (more than 10 species) mainlyin open areas of nitrogen-rich soils and water bodies

It is surprising that pollen showing the highest proteincontent (gt26) belonged to highly nectariferous plants wellrepresented inmeliponine andApis honey in this region suchas Prosopis Maytenus and Ziziphus [39 41ndash45] Studies onprotein content of pollen loads and pollen analysis of honeycarried out in other semiarid areas also show this tendencyas seen in Table III of [35] and Table II of [46] for CondaliaBrassicaceae and Prosopis in Table III of [36] and in Table IIof [47] for Adesmia Rosaceae Trifolium Melilotus Schinusand Brassicaceae Furthermore pollen from the nectariferousLarrea and Prosopis showed the richest protein value in astudy carried out in a North American desert [38] Thispattern of high protein value in pollen of highly nectariferousplants foraged by honey-producing bees is here hypothesizedand should be further studied

Conflict of Interests

The author declares that there is no conflict of interestsregarding the publication of this paper

Acknowledgments

The author thanks Cesar Albornoz InocencioMedina IsabelBrait Rogelio Burgardt Mercedes Koler Ricardo ldquoNenerdquoVossler and Juan Hiperdinger for their warm hospitality andhelp during the field studies in El Sauzalito J J Castelli ElEspinillo and Villa Rıo Bermejito and Nora Brea for pro-viding suggestions and comments on the paper The authoris especially grateful to Arturo Roig-Alsina for identifyingthe bees This study was supported by CONICET (ConsejoNacional de Investigaciones Cientıficas y Tecnicas)

References

[1] D W Roubik Ecology and Natural History of Tropical BeesCambridge University Press New York NY USA 1989

[2] M Ramalho M D Silva and C A L Carvalho ldquoDinamicade uso de fontes de polen por Melipona scutellaris Latreille(Hymenoptera Apidae) uma analise comparativa com Apis

6 Psyche

mellifera L (Hymenoptera Apidae) no Domınio TropicalAtlanticordquo Neotropical Entomology vol 36 no 1 pp 38ndash452007

[3] DW Roubik ldquoStingless bee nesting biologyrdquoApidologie vol 37no 2 pp 124ndash143 2006

[4] D B Casteel ldquoThe behaviour of the honeybee in pollen col-lectingrdquo Bulletin of the United States Department of AgricultureBureau of Entomology vol 121 pp 1ndash36 1912

[5] M H Haydak ldquoPollen substitutesrdquo in Proceedings of the 10thInternational Congress of Entomology vol 4 pp 1053ndash1055Montreal Canada August 1958

[6] T C Olofsson and A Vasquez ldquoDetection and identification ofa novel lactic acid bacterial flora within the honey stomach ofthe honeybee Apis melliferardquo Current Microbiology vol 57 no4 pp 356ndash363 2008

[7] A Vasquez and T C Olofsson ldquoThe lactic acid bacteriainvolved in the production of bee pollen and bee breadrdquo Journalof Apicultural Research vol 48 no 3 pp 189ndash195 2009

[8] M Gilliam D W Roubik and B J Lorenz ldquoMicroorganismsassociated with pollen honey and brood provisions in the nestof a stingless bee Melipona fasciatardquo Apidologie vol 21 no 2pp 89ndash97 1990

[9] A Vasquez E Forsgren I Fries et al ldquoSymbionts as majormodulators of insect health lactic acid bacteria and honeybeesrdquoPLoS ONE vol 7 no 3 Article ID e33188 2012

[10] D W Roubik ldquoSeasonality in colony food storage broodproduction and adult survivorship studies of Melipona intropical forest (Hymenoptera Apidae)rdquo Journal of the KansasEntomological Society vol 55 pp 789ndash800 1982

[11] F B Noll ldquoForaging behavior on carcasses in the necrophagicbee Trigona hypogea (Hymenoptera Apidae)rdquo Journal of InsectBehavior vol 10 no 3 pp 463ndash467 1997

[12] H F Schwarz ldquoStingless bees (Meliponidae) of the WesternHemisphererdquo Bulletin of the American Museum of NaturalHistory vol 90 pp 1ndash546 1948

[13] DWRoubik ldquoObligate necrophagy in a social beerdquo Science vol217 no 4564 pp 1059ndash1060 1982

[14] JM F Camargo andDW Roubik ldquoSystematics and bionomicsof the apoid obligate necrophages the Trigona hypogea group(Hymenoptera Apidae Meliponinae)rdquo Biological Journal of theLinnean Society vol 44 no 1 pp 13ndash39 1991

[15] C Rasmussen and J M F Camargo ldquoA molecular phylogenyand the evolution of nest architecture and behavior in Trigonass (Hymenoptera ApidaeMeliponini)rdquoApidologie vol 39 no1 pp 102ndash118 2008

[16] D C Somerville ldquoNutritional value of bee collected pollenrdquoReport for the Rural Industries Research and DevelopmentCorporation Rural Industries Research and Development Cor-poration Barton Australia 2001

[17] G J Kleinschmidt and A C Kondos ldquoInfluence of crude pro-tein levels on colony productionrdquo The Australasian Beekeepervol 78 pp 36ndash39 1976

[18] E J Slaa L A S Chaves K S Malagodi-Braga and F EHofstede ldquoStingless bees in applied pollination practice andperspectivesrdquo Apidologie vol 37 no 2 pp 293ndash315 2006

[19] M Cortopassi-Laurino V L Imperatriz-Fonseca DW Roubiket al ldquoGlobal meliponiculture challenges and opportunitiesrdquoApidologie vol 37 no 2 pp 275ndash292 2006

[20] F G Vossler ldquoFlower visits nesting and nest defence behaviourof stingless bees (Apidae Meliponini) suitability of the beespecies for meliponiculture in the Argentinean Chaco regionrdquoApidologie vol 43 no 2 pp 139ndash161 2012

[21] A Roig-Alsina F G Vossler and G P Gennari ldquoStingless beesin Argentinardquo in Pot Honey A Legacy of Stingless Bees P Vit SRM Pedro andDW Roubik Eds pp 125ndash134 Springer NewYork NY USA 2013

[22] P Arenas Etnografıa y Alimentacion Entre los Toba-Nachilamoleek yWichı Lhukursquotas del Chaco Central (Argentina)Pastor Arenas Publisher Buenos Aires Argentina 2003

[23] A L Cabrera ldquoFitogeografıa de la RepublicaArgentinardquoBoletınde la Sociedad Argentina de Botanica vol 14 pp 1ndash42 1971

[24] D E Prado ldquoWhat is the Gran Chaco vegetation in SouthAmerica I A review Contribution to the study of flora andvegetation of the Chacordquo Candollea vol 48 pp 145ndash172 1993

[25] G Erdtman ldquoThe acetolysis method a revised descriptionrdquoSvensk Botanisk Tidskrift vol 54 pp 561ndash564 1960

[26] F O Zuloaga O Morrone and M J Belgrano Catalogo delas Plantas Vasculares del Cono Sur (Argentina Sur de BrasilChile Paraguay y Uruguay) vol 107 Monographs in SystematicBotany from the Missouri Botanical Garden 2008

[27] AOAC (Association of Official Analytical Chemists) OfficialMethods of Analysis AOAC Washington DC USA 1980

[28] J M Bremner and C S Mulvaney ldquoNitrogen-totalrdquo inMethodsof Soil Analysis Part 2 Chemical andMicrobiological PropertiesA L Page R H Miller and D R Keeney Eds pp 595ndash624American Society of Agronomy Madison Wis USA 1982

[29] T H Roulston and J H Cane ldquoPollen nutritional content anddigestibility for animalsrdquo Plant Systematics and Evolution vol222 no 1ndash4 pp 187ndash209 2000

[30] EWHerbert ldquoHoney bee nutritionrdquo inTheHive and the HoneyBee J E Graham Ed pp 197ndash233 Dadant amp Sons HamiltonIll USA 1992

[31] J Adamoli E Sennhauser J M Acero and A Rescia ldquoStressand disturbance vegetation dynamics in the dry Chaco regionof Argentinardquo Journal of Biogeography vol 17 no 4-5 pp 491ndash500 1990

[32] S D Leonhardt and M Kaltenpoth ldquoMicrobial communitiesof three sympatric Australian stingless bee speciesrdquo PLoS ONEvol 9 no 8 Article ID e105718 2014

[33] E Butler M Alsterfjord T C Olofsson C Karlsson J Malm-strom and A Vasquez ldquoProteins of novel lactic acid bacteriafrom Apis mellifera mellifera an insight into the production ofknown extra-cellular proteins during microbial stressrdquo BMCMicrobiology vol 13 article 235 2013

[34] J H Cane and S Sipes ldquoCharacterizing floral specialization bybees analytical methods and a revised lexicon for oligolectyrdquo inPlant-Pollinator Interactions from Specialization to Generaliza-tion N M Waser and J Ollerton Eds pp 99ndash122 Universityof Chicago Press Chicago Ill USA 2006

[35] A C Andrada and M C Tellerıa ldquoPollen collected byhoney bees (Apis mellifera L) from south of Calden district(Argentina) botanical origin and protein contentrdquo Grana vol44 no 2 pp 115ndash122 2005

[36] A Forcone P V Aloisi S Ruppel and M Munoz ldquoBotanicalcomposition and protein content of pollen collected by Apismellifera L in the north-west of Santa Cruz (ArgentineanPatagonia)rdquo Grana vol 50 no 1 pp 30ndash39 2011

[37] F E Todd and O Bretherick ldquoThe composition of pollensrdquoJournal of Economic Entomology vol 35 no 3 pp 312ndash317 1942

[38] W FMcCaughey M Gilliam and L N Standifer ldquoAmino acidsand protein adequacy for honey bees of pollens from desertplants and other floral sourcesrdquoApidologie vol 11 no 1 pp 75ndash86 1980

Psyche 7

[39] F G Vossler M C Tellerıa and M Cunningham ldquoFloralresources foraged byGeotrigona argentina (ApidaeMeliponini)in the Argentine Dry Chaco forestrdquo Grana vol 49 no 2 pp142ndash153 2010

[40] T H Roulston J H Cane and S L Buchmann ldquoWhat governsprotein content of pollen pollinator preferences pollen-pistilinteractions or phylogenyrdquo Ecological Monographs vol 70 no4 pp 617ndash643 2000

[41] A Basilio and M Noetinger ldquoAnalisis polınico de mieles de laRegion Chaquena comparacion del origen floral entre las zonasDomoCentral y Esteros Canadas y Selvas de RiverardquoRevista deInvestigaciones Agropecuarias vol 31 no 2 pp 127ndash134 2000

[42] M Cabrera ldquoCaracterizacion polınica de las mieles de laprovincia de Formosa Argentinardquo Revista del Museo Argentinode Ciencias Naurales Bernardino Rivadavia NS vol 8 pp 135ndash142 2006

[43] C R Salgado Floramelıfera en la provincia del Chaco Edited byPROSAP Impreso por Ministerio de la Produccion del Chaco2006

[44] A M Basilio C Spagarino L Landi and B Achaval ldquoMielde Scaptotrigona jujuyensis en dos localidades de FormosaArgentinardquo in Stingless Bees Process Honey and Pollen inCerumen Pots P Vit and D W Roubik Eds pp 1ndash8 Facultadde Farmacia y Bioanalisis Universidad de Los Andes MeridaVenezuela 2013 httpwwwsaberulavehandle1234567893529

[45] F G Vossler G A Fagundez and D C Blettler ldquoVariabilityof food stores of Tetragonisca fiebrigi (Schwarz) (HymenopteraApidae Meliponini) from the Argentine Chaco based on pollenanalysisrdquo Sociobiology vol 61 no 4 pp 449ndash460 2014

[46] A C Andrada and M C Tellerıa ldquoBotanical origin of honeyfrom south of Calden district (Argentina)rdquoGrana vol 41 no 1pp 58ndash62 2002

[47] A Forcone G Ayestaran A Kutschker and J Garcıa ldquoPalyno-logical characterization of honeys from the Andean Patagonia(Chubut Argentina)rdquo Grana vol 44 no 3 pp 202ndash208 2005

Submit your manuscripts athttpwwwhindawicom

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Anatomy Research International

PeptidesInternational Journal of


Hindawi Publishing Corporation httpwwwhindawicom

International Journal of

Volume 2014

Zoology


Molecular Biology International

GenomicsInternational Journal of


The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014


BioinformaticsAdvances in

Marine BiologyJournal of



Signal TransductionJournal of


BioMed Research International

Evolutionary BiologyInternational Journal of



Biochemistry Research International

ArchaeaHindawi Publishing Corporationhttpwwwhindawicom Volume 2014


Genetics Research International


Advances in

Virolog y

Hindawi Publishing Corporationhttpwwwhindawicom

Nucleic AcidsJournal of

Volume 2014

Stem CellsInternational



Enzyme Research



Microbiology

2 Psyche

amount required by colonies for optimum production [17]A well-nourished bee colony is a guarantee for good level ofhoney flow and breeding [16]

It is important to know pollen protein intake of melipo-nine bees for appropriate colony rearing (meliponiculture)as it would be useful to determine supplemental feedingneeded in periods of flowering shortage Stingless bees areimportant for pollination of wild and crop plants [18] and forbee-products production (honey pollen resin and cerumen)[19] The aims of the present study were to determine proteincontent of pollen stored in nests of three meliponine beespecies (Tetragonisca fiebrigi (Schwarz) Melipona orbignyi(Guerin) andGeotrigona argentinaCamargo ampMoure) fromthe South American Chaco dry forest These are three outof the seven species recorded for this area [20 21] beingimportant mainly for rural population for their honey pollenmasses larvae and cerumen [22]

2 Materials and Methods

21 Sampling and Study Area Pollen mass from ceru-men pots was obtained from nests of Tetragonisca fieb-rigi (Schwarz) Melipona orbignyi (Guerin) and Geotrigonaargentina Camargo amp Moure from xeric forests in the Chacoregion of Northern Argentina (Figure 1) Samples for proteinanalysis were taken from nest 7 of T fiebrigi nest 11 ofM orbignyi and nests 2 and 4 of G argentina from ElSauzalito (24∘241015840S 61∘401015840W) from nest 1 of G argentinafrom El Espinillo (25∘ 241015840S 60∘ 27W) from nest 5 ofT fiebrigi from Miraflores (25∘291015840S 61∘011015840W) and fromnest 12 of T fiebrigi from Villa Rıo Bermejito (25∘371015840S60∘151015840W) This Chaco dry forest is characterized by thedominance of ldquopalo santordquo (Bulnesia sarmientoi Lorentz exGriseb Zygophyllaceae) and ldquoquebracho blancordquo trees (Aspi-dosperma quebracho-blanco Schltdl Apocynaceae) or byldquoquebracho colorado santiaguenordquo (Schinopsis lorentzii EnglAnacardiaceae) ldquoquebracho colorado chaquenordquo (Schinopsisbalansae Engl) and ldquoquebracho blancordquo [23 24] Otherwoody elements well represented in this xerophilous flora areldquomistolrdquo (Ziziphus mistol Griseb Rhamnaceae) ldquomollerdquo orldquoguaraninardquo (Sideroxylon obtusifolium (Roem amp Schult) TD Penn Sapotaceae) several species of Prosopis (ldquoalgarroboblancordquo ldquoalgarrobo negrordquo ldquovinalrdquo ldquovinalillordquo ldquocarandardquoand ldquoguachınrdquo) (Fabaceae Mimosoideae) ldquoguayacanrdquo (Cae-salpinia paraguariensis (D Parodi) Burkart Fabaceae Cae-salpinioideae) ldquotipa coloradardquo or ldquopalo cocardquo (Pterog-yne nitens Tul Fabaceae Caesalpinioideae) ldquopalo cruzrdquo(Tabebuia nodosa (Griseb) Griseb Bignoniaceae) ldquoque-brachillordquo (Aspidosperma triternatum Rojas Acosta Apocy-naceae) ldquopalo borrachordquo or ldquoyuchanrdquo (Ceiba chodatii (Hassl)Ravenna Bombacaceae) ldquopalma de monterdquo (Trithrinaxschizophylla Drude Arecaceae) ldquomeloncillordquo (Castela coc-cinea Griseb Simaroubaceae) ldquopalo tintardquo (Achatocarpuspraecox Griseb Achatocarpaceae) ldquotalardquo (Celtis spp Celti-daceae) ldquoduraznillordquo (Salta triflora (Griseb) Adr SanchezPolygonaceae) ldquopatardquo (Ximenia americana L Olacaceae)ldquomollerdquo (Schinus fasciculatus (Griseb) I M Johnst vararenicola (Hauman) F A Barkley Anacardiaceae) ldquochanarrdquo

(Geoffroea decorticans (Gillies ex Hook amp Arn) BurkartFabaceae Papilionoideae) ldquosal de indiordquo or ldquosal de monterdquo(Maytenus vitis-idaea Griseb Celastraceae) ldquopaloma yuyordquo(Moya spinosa Griseb Celastraceae) ldquoteatınrdquo (Mimosa deti-nens Benth Fabaceae Mimosoideae) ldquocardonrdquo (Stetsoniacoryne (Salm-Dyck) Britton amp Rose Cactaceae) ldquouclerdquo(Cereus forbesii Otto ex C F Forst Cactaceae) twospecies of Bougainvillaea (ldquorama overardquo) (Nyctaginaceae)several species of Acacia (ldquotuscardquo ldquogarabatordquo and ldquochurquirdquo)(Fabaceae Mimosoideae) Capparis (ldquoatamisquirdquo ldquosachamembrillordquo ldquosacha sandiardquo ldquosacha porotordquo and ldquobolaverderdquo) and ldquocardordquo or ldquochaguarrdquo (several Bromeliaceaegenera) among others

22 Pollen Analysis of Pollen Masses and Plant and BeeReferences Pollen masses were dissolved in distilled water at80ndash90∘C and stirred with a magnetic stirrer for 10ndash15minA representative mixture of 5ndash10mL was obtained and cen-trifuged at 472timesg for 5min Processing included acetolysis[25] Under a Nikon Eclipse E200 light microscope a total of300ndash500 pollen grains per slide were counted Pollen grainidentification was carried out comparing nest pollen slideswith those present in the reference pollen collection Thereference slides consist of a total of 190 plant species and aredeposited in PAL-CICYTTP pollen collection of DiamanteEntre Rıos Argentina It was made from flower buds of plantspecies collected in various localities from Chaco Province ofArgentina These plant specimens were pressed dried andidentified by the author and deposited in the Herbariumof the Museo de La Plata (LP) the Herbarium of MuseoArgentino de Ciencias Naturales ldquoBernardino Rivadaviardquo(BA) Buenos Aires and the Herbarium Lorentz (DTE) ofDiamante Entre Rıos Argentina Plant nomenclature follows[26] Bee specimens were collected from nests identifiedby Arturo Roig-Alsina and deposited in the EntomologyCollection of the Museo Argentino de Ciencias NaturalesldquoBernardino Rivadaviardquo Buenos Aires Argentina

23 Determination of Protein Content To quantify proteinof pollen resources foraged by bees certain pollen masseswere selected according to their pollen type compositionSix pollen mass samples were taken from nests 5 7 and12 of Tetragonisca fiebrigi one from nest 11 of Meliponaorbignyi and eight from nests 1 2 and 4 of Geotrigonaargentina (Table 1) For nitrogen content determination50mg of pollen [27] was analyzed by the micro-Kjeldahlmethod [28] and crude protein was estimated using thefactor 625 [29] Pollen nitrogen content was analyzed inLANAIS N15 (National Laboratory of Research and ServicesUNS-CONICET) Departamento de Agronomıa Universi-dad Nacional del Sur Bahıa Blanca Argentina

3 Results

31 Protein Content of Pollen Stored in Meliponine NestsNitrogen values from pollen grains stored in pollen potsof the three meliponine species studied varied from 156to 486 which is equivalent to 978 to 3041 of crude

Psyche 3

Table1Mainpo


n)presentinpo

llenmasssam

ples

ofTetra

goniscafiebrigi(Tf

)Melipona

orbignyi(M

o)and

Geotrigona

argentina(G

a)andtheirn

itrogen

and

proteinpercentages

Bees

pecies

Sample

Nestand

pollenmass

Mainpo

llentypes

Nitrogen

()

Crud

eprotein

()

Tf

15B

Type

Schinopsis(59

)+Trith

rinax

schizophylla(405

)3055

1909

25C

Trith

rinax

schizophylla(41

)+Pa

rthenium

hyste

rophorus

(30

)+type

Schinopsis(28

)2854

1784

35E

Trith

rinax

schizophylla(98

)2898

1811

47A


(24

)+Prosopis(18

)+type

Maytenu

svitis-idaea

(16

)+Tabebuia(13

)3126

1954

57F

Capparisspeciosa

(42


(34

)+Heliantheae

(10

)364

82280

612

CZizip

husm

istol(60

)+Prosopis(30

)3542

2214

Mo

711

Type

Acaciapraecox(53

)+Prosopis(38

)3881

2426

Ga

8458

Type

Capparistweedian

a-C

speciosa

(100)

156

978

912

4Ca

stelacoccinea

(100)

326

2036

10231

Prosopis(100)

334

2087

11111

Prosopis(91

)486

3041

12236

Prosopis(895

)

432

2703

13233

Type

Maytenu

svitis-idaea

(100)

423

2687

14242

Type

Maytenu

svitis-idaea

(96

)424

2651

15116

Type

Croton

(30

)+type

Sagitta

ria(18

)+Eleocharis(12

)+Ca

stelacoccinea

(11

)265

1659

4 Psyche

(a) (b) (c)

(d) (e) (f)




4 Discussion



Psyche 5








Acknowledgments


References



6 Psyche






































Psyche 7

















Volume 2014

Zoology






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology

Psyche 3

Table1Mainpo


n)presentinpo

llenmasssam

ples

ofTetra

goniscafiebrigi(Tf

)Melipona

orbignyi(M

o)and

Geotrigona

argentina(G

a)andtheirn

itrogen

and

proteinpercentages

Bees

pecies

Sample

Nestand

pollenmass

Mainpo

llentypes

Nitrogen

()

Crud

eprotein

()

Tf

15B

Type

Schinopsis(59

)+Trith

rinax

schizophylla(405

)3055

1909

25C

Trith

rinax

schizophylla(41

)+Pa

rthenium

hyste

rophorus

(30

)+type

Schinopsis(28

)2854

1784

35E

Trith

rinax

schizophylla(98

)2898

1811

47A


(24

)+Prosopis(18

)+type

Maytenu

svitis-idaea

(16

)+Tabebuia(13

)3126

1954

57F

Capparisspeciosa

(42


(34

)+Heliantheae

(10

)364

82280

612

CZizip

husm

istol(60

)+Prosopis(30

)3542

2214

Mo

711

Type

Acaciapraecox(53

)+Prosopis(38

)3881

2426

Ga

8458

Type

Capparistweedian

a-C

speciosa

(100)

156

978

912

4Ca

stelacoccinea

(100)

326

2036

10231

Prosopis(100)

334

2087

11111

Prosopis(91

)486

3041

12236

Prosopis(895

)

432

2703

13233

Type

Maytenu

svitis-idaea

(100)

423

2687

14242

Type

Maytenu

svitis-idaea

(96

)424

2651

15116

Type

Croton

(30

)+type

Sagitta

ria(18

)+Eleocharis(12

)+Ca

stelacoccinea

(11

)265

1659

4 Psyche

(a) (b) (c)

(d) (e) (f)




4 Discussion



Psyche 5








Acknowledgments


References



6 Psyche






































Psyche 7

















Volume 2014

Zoology






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology

4 Psyche

(a) (b) (c)

(d) (e) (f)




4 Discussion



Psyche 5








Acknowledgments


References



6 Psyche






































Psyche 7

















Volume 2014

Zoology






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology

Psyche 5








Acknowledgments


References



6 Psyche






































Psyche 7

















Volume 2014

Zoology






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology

6 Psyche






































Psyche 7

















Volume 2014

Zoology






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology

Psyche 7

















Volume 2014

Zoology






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology








Volume 2014

Zoology






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology

research article broad protein spectrum in stored pollen...

Documents