Millipedes as Food for Humans: Their Nutritional and Possible Antimalarial Value—A First Report

Hindawi Publishing CorporationEvidence-Based Complementary and Alternative MedicineVolume 2014 Article ID 651768 9 pageshttpdxdoiorg1011552014651768

Research ArticleMillipedes as Food for Humans Their Nutritional andPossible Antimalarial ValuemdashA First Report

Henrik Enghoff1 Nicola Manno23 Seacuteveacuterin Tchibozo4

Manuela List5 Bettina Schwarzinger5 Wolfgang Schoefberger6

Clemens Schwarzinger5 and Maurizio G Paoletti2

1 Natural History Museum of Denmark University of Copenhagen Universitetsparken 152100 Copenhagen Oslash Denmark

2Dipartimento di Biologia Universita di Padova lab Agroecology and EthnobiologyVia U Bassi 58b 35121 Padova Italy

3 Escuela de Postgrado en Ciencias Biologicas Universidad Nacional de Trujillo Peru4Centre de Recherche pour la Gestion de la Biodiversite 04 BP 0385 Cotonou Benin5 Institute for Chemical Technology of Organic Materials Johannes Kepler University LinzAltenberger Strasse 69 4040 Linz Austria

6 Institute for Inorganic Chemistry Johannes Kepler University Linz Altenberger Strasse 69 4040 Linz Austria

Correspondence should be addressed to Maurizio G Paoletti paoletticivbiounipdit

Received 15 May 2013 Revised 22 October 2013 Accepted 23 October 2013 Published 29 January 2014

Academic Editor Tung-Sheng Chen

Copyright copy 2014 Henrik Enghoff et alThis is an open access article distributed under the Creative Commons Attribution Licensewhich permits unrestricted use distribution and reproduction in any medium provided the original work is properly cited

The first record of millipedes (Diplopoda) being regularly used for food by humans (the Bobo people of Burkina Faso) is givenincluding information on how the millipedes are prepared The species in question are Tymbodesmus falcatus (Karsch 1881) andSphenodesmus sheribongensis (Schioslashtz 1966) (Gomphodesmidae) and an unidentified species of Spirostreptidae New informationon the nutritional value of millipedes is provided unsaturated fatty acids calcium and iron contents are particularly high Themillipedesrsquo defensive secretions hydrogen cyanide and benzoquinones present a severe challenge for the spread of millipedes as aneveryday food source On the other hand the possibility that benzoquinones may act as insect-repellents as known from studieson nonhuman primates and that sublethal cyanide ingestion may enhance human innate resistance to malaria suggests promisingethnomedical perspectives to our findings

1 Introduction

Small vertebrates and invertebrates especially insects theso called minilivestock have been considered a promisingresource for Earthrsquos human population that will reach 9billion humans in 2050 [1ndash3] and are potential candidatesfor reducing the higher and increasing impact on resourcesrepresented by larger livestock and inland fish production [4]Information on traditional local use of these small animalsis an important starting point for studying minilivestockas potential food resources for humans [5ndash8] In additionlocal use of invertebrates may have unexpected ethnomedicalimplications

Millipedes (Diplopoda) have so far not been in focusas minilivestock Indeed most orders of millipedes (Glom-erida Polyzoniida Siphonocryptida Platydesmida Siphono-phorida Callipodida Julida Spirobolida Spirostreptida andPolydesmida) are known for their chemical defenses andunlike their relatives the centipedes (Chilopoda) which inseveral cultures (China Alto Orinoco in Venezuela andKorea) have been used as medical remedies andor fooditems [9 10] no information on millipedes as human foodhas been available until now A wide spectrum of chemicalshas been identified from millipede defensive secretions [11]the most widespread ones being benzoquinones (in mostcylindrical millipedes superorder Juliformia) and hydrogen

2 Evidence-Based Complementary and Alternative Medicine

cyanide derived frommandelonitrile and related compounds(inmost flatbackedmillipedes superorderMerocheta)Thesetoxic smelly chemicals make millipedes unattractive formost predators although there are some animals vertebratesas well as invertebrates which eat millipedes [12 13] andsome are even specialized on a millipede diet for exampleassassin bugs (family Reduviidae) of the subfamily Ectri-chodiinae [14] and beetle larvae of the family Phengodidae[15] A few vertebrates are reported to eat millipedes asfor instance banded mongoose (Mungos mungo) [16] Somebirds and nonhominid primates use toxic millipedes forldquoself-anointmentrdquo presumably exploiting an insect-repellenteffect of the millipedesrsquo defensive chemicals especially ben-zoquinones [17ndash20]We have however not been able to traceany record of millipedes being used as food in any humansociety but nowwe can report the consumption ofmillipedesby the Bobo population of Burkina Faso a region whereentomophagy has been extensively described [21]

Themillipedeswhich are used as human food by the Bobobelong to two families Gomphodesmidae and Spirostrepti-dae

Gomphodesmidae (flatbacked millipedes of the orderPolydesmida) (Figures 1 and 2) belong to the ldquocyanogenicrdquomillipedes the family is endemic to the African continentsouth of the Sahara includes 146 named species [22] andwas monographed by Hoffman (2005) [23] The only gom-phodesmid species previously recorded from Burkina Fasois Tymbodesmus falcatus (Karsch 1881) collected in Oua-gadougou [23] One of the gomphodesmid species used forfood at Kou village in Burkina Faso is indeed T falcatus (DVandenspiegel det) the other is Sphenodesmus sheribongensis(Schioslashtz 1966) (HE det) T falcatus is known from MaliBurkina Faso Nigeria Sudan and Central African RepublicS sheribongensis was previously known from Ghana IvoryCoast andNigeria [23] Lewis [24] studied the life history andecology of both species in Zaria NorthernNigeria Both havea two-year life cycle Juvenile stadia of S sheribongensis liveentirely in the soil whereas adults can be extremely abundanton the soil surface during the first part of the rainy season(from May to July) T falcatus is similar except that the lastjuvenile (subadult) stadium is seasonally surface active likethe adults albeit in smaller numbers

Spirostreptidae (cylindrical millipedes of the order Spiro-streptida) (Figure 3) belong to the ldquoquinonerdquo millipedes thefamily is near-endemic to the Afrotropical and Neotropicalregions includes 275 named species [22] and was mono-graphed by Krabbe [25] No species of Spirostreptidae havebeen recorded from Burkina Faso and the one occurring atKou has not yet been identified

We do not have any strong evidence that T falcatus andS sheribongensis actually produce hydrogen cyanide nor thatthe spirostreptid in question produces quinones The generaloccurrence of these substances in the respective higher taxato which the species belong however is strong circumstantialevidence that this is actually so although recent studieshave demonstrated a larger diversity in millipede defensivechemicals than previously assumed [26ndash28]

Until this study the nutritional value of millipedes hasnever been assessed Considering that the muscle volume

Figure 1 Live gomphodesmid millipede (Tymbodesmus falcatus)from Kou S Tchibozo phot 2011

Figure 2 gomphodesmid millipede from Kou after boiling S Tchi-bozo phot 2012

Figure 3 Spirostreptidmillipede fromKou S Tchibozo phot 2012

of millipede is small they will constitute a poor source ofproteinTheir gutsmainly contain soil and litter remains [24]but their calcified exoskeletonmight constitute a considerablesource of calcium which may constitute 13ndash17 of the dryweight [29ndash31] or 9 of the fresh weight [32] In factmillipedes are considered an essential source of calcium foregg production in certain birds [30 33 34] In this workwe present data based on Tymbodesmus falcatus one of thespecies eaten by the Bobo people

2 Material and Methods

21 Sampling and Ethnobiological Data Collection Obser-vations and interviews with the Bobo people were made

Evidence-Based Complementary and Alternative Medicine 3

by ST in 2011 and 2012 in Burkina Faso (Kou 11∘10881015840N004∘26621015840W altitude 351m near Bobo-Dioulasso) Exem-plars of the edible millipedes were collected and are nowpreserved in Musee Royale de lrsquoAfrique Centrale (TervurenBelgium) and the Natural History Museum of Denmark(Copenhagen) Five Bobo people especially women wereinterviewed for information on collection and preparation ofthe edible millipedes

22 Nutritional Analysis In order to assess the nutritionalvalue of this unconventional food we determined variousnutritional parameters of a whole Tymbodesmus falcatusmale specimen The raw specimen was preserved in 70ethanol subsequently dried and homogenized by cryomilling(SPEX FreezerMill 6770) The resulting powder was usedfor analysis of chitin fatty acids amino acids and metalcontent No appropriately preserved specimens of the otherspecies in question were available Dry weight of millipedeswas measured on oven-dried specimens and kept in alcoholfor 45 years (collected in Nigeria by JGE Lewis)

221 Pyrolysis-GCMS for Fatty Acid Analysis 100 120583g of thesample was placed in a quartz tube and 45 120583L of a dilutedaqueous solution of tetramethylammonium hydroxide wasadded The samples were subsequently pyrolyzed at 450∘Cfor 10 s with a CDS 5250 pyrolysis autosampler attached to aThermo Trace GC UltraMD 800 gas chromatographymassspectrometry system Volatile products were separated ona Supelco SP 2330 column (30m ID 032mm 02120583m filmthickness) with helium 46 as carrier gas (2mLsdotminminus1) andidentified by comparison to reference compounds as well asinterpretation of their EI mass spectra and comparison toNIST 2002Wiley and NBS electronic librariesThe pyrolysisinterfacewas kept at 300∘Cand theGCMS interface at 280∘CtheGCwas programmed from 100∘C (1min) to 230∘C (5min)at a rate of 10∘C minminus1 The mass spectrometer was operatedin EI mode (70 eV) at a source temperature of 200∘C [35]

222 Solid State NMR for Chitin Chitin was determined bySolid State NMR all spectra were recorded on a narrow-bore117 T instrument (500MHz 1H Larmor frequency) at magicangle spinning rates of 100 kHz at 300K 13C chemical shiftsare given in reference to tetramethylsilane TMS using thesharp resonance of TMS as external calibration A basic crosspolarization experiment with total suppression of sidebandswith a cross-polarization contact time of 2ms was employedwith an effective acquisition time of 279ms and a recyclingdelay of 5 s The magic angle was adjusted using the 79Brresonance of KBr and the actual sample temperature wasdetermined using the 207Pb resonance of Pb(NO

3)2for calib-

ration [36]

223 Amino Acid Analysis The powder obtained after cry-omilling was hydrolysed by refluxing with hydrochloric acidcontaining 5 phenol for 24 hours under exclusion ofoxygenThe complete samplewas then evaporated to drynessredissolved in water and analyzed with HPLCMS using05mLsdotminminus1 of a water acetonitrile gradient (100 water

Table 1 Tymbodesmus falcatus amino acid analysis (based on drymatter)

Amino acid mgmgAlanin 00676Asparagin 00137Asparaginic acid 00164Glutamin + lysin 00217Glutamic acid 00000Glycin 00000Isoleucin + leucin 00283Methionin 00056Phenylalanin 00132Prolin 00062Serin 00142Threonin 00251Tryptophan 00053Tyrosin 00151Valin 00193Total 02518

for 2 minutes and in 17 minutes to 30 acetonitrile which isheld for a further 3 minutes) on a Waters AccQ Tag column(39 times 150mm) Quantification was done using extracted ionchromatograms With this procedure arginine cysteine andhistidine could not be analyzed and leucineisoleucine as wellas glutaminelysine could not be separated therefore theconcentration of those amino acids is given as a sum

224 Metal Analysis Metal content was analysed withinductively coupled plasma optical emission spectrometry(ICP-OES) according to EN ISO 11885 from a commerciallaboratory

3 Results

31 Collection and Preparation of Millipedes at Kou VillageAccording to the interviewed villagers in Kou millipedesare collected under bricks around houses made of straw andunder decomposing wood Once collected the millipedes areplaced in a pot with water filtered through firewood ashesfor 3ndash5 minutes until boiling Then they are removed andleft to dry on a roof for 3 days Such preparation is specificfor millipedes and different from those described for otherarthropods in West Africa especially for insect maggots andweevils which are mainly roasted and fried [21] The driedmillipedes are placed in a tomato sauce to which is added thetraditional African mustard known as soumbala (fermentedseeds of the nere tree Parkia biglobosa very widely consumedin Burkina Faso and West Africa in general) shea butter oiland to (a pastemade frommaize or sorghum flour) For somemeals the millipedes replace meat

32 Nutritional Values of Tymbodesmus falcatus Proteinsrepresent 25 of total dry weight (calculated as the sum ofamino acids) the amino acid profile (Table 1) is similar to that


Table 2 Fatty acid distribution as determined by pyrolysis-GCMSanalysis Unsaturated fatty acids constitute 40 of total fatty acids ofTymbodesmus falcatus

No ID Name 119905119877

Percent1 100 Caprylic acid 3 032 120 Lauric acid 433 053 140 Myristic acid 582 314 150 Pentadecanoic acid 656 105 160 Palmitic acid 737 4316 161 n9 Sapienic acid 761 027 161 n7 Palmitoleic acid 767 158 170 Margaric acid 801 089 180 Stearic acid 873 7810 181 n9 Oleic acid 906 36811 181 n7 Vaccenic acid 0012 182 n6 Linoleic acid 95 1613 183 n6 Gamma linolenic acid 0014 200 Arachidic acid 10 1115 183 n3 Alpha linolenic acid 1008 0116 182 x Octadecenoic acid 1024 0517 201 n9 Gadoleic acid 0018 182 x Octadecenoic acid 1034 0519 182 x Octadecenoic acid 1055 0320 202 Eicosadienoic acid 0021 203 Eicosatrienoic acid 0022 220 Behenic acid 112 0123 204 n6 Arachidonic acid 0024 210 Heneicosylic acid 1177 0125 205 n3 Timnodonic acid 0026 240 Lignoceric acid 1233 0427 225 n6 Docosapentaenoic acid 0028 226 n3 Docosahexaenoic acid 00Total 1000

of insects and crustaceans for example crickets and shrimps[37] Unsaturated fatty acids constitute a relevant fraction(40) of total fatty acids (Table 2 Figure 4) however lowerthan those described for widely consumed and appreciatededible insects [38] Calcium levels (Table 3) are very high(174 of dry weight) which is higher than previously pub-lished values [29ndash32]

The dry weight of individual Tymbodesmus falcatus was042ndash054 g (mean 046 119899 = 3) and of the smaller Sphen-odesmus sheribongensis 008ndash011 g (mean 009 119899 = 4)Spirostreptidae vary very much in size live weights of upto 80 g have been measured (HE unpublished) Consideringthat the Ca content of a single Tymbosdesmus f is about80ndash90mg (174mgg times 046 g) consumption of around 12-13 gomphodesmid individuals will provide 1000mgday (theDietary Reference Intake (DRI) by IOM 2004 [39]) Alsoiron content (100600mgkg) is important considering thatonly 6 individuals provide the adequate amount for a womenduring pregnancy (DRI 27mgday) and only 2-3 for men(DRI 8mgday) [39]

0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18Time (min)

0102030405060708090

100 737906

873

582 17153

4 7

10

9

5

1214

NN

Rela

tive a

bund

ance

Figure 4 THM-GCMS profile of Tymbodesmus falcatus retentiontimes (119905

119877

) refer to compounds listed in Table 2 palmitic acid 18 1n9 (119905119877

737) (5) and oleic acid 16 0 (119905119877

906) (10) constitute 80 oftotal fatty acids

Table 3 Tymbodesmus falcatus metal contents (based on drymatter) and DRIs for a pregnant women of 19ndash30 yrs by IOM 2004[39]

Metal mgkg DRI (mgday)Pb 62 mdashCd lt05 mdashCa 174000 1000Fe 10600 27K 2610 4700Cu 789 1Mg 4990 350Na 1630 1500Zn 160 11

Chitin constitutes around 5 of the total dry weight alow amount compared to Ca but it is only located in theexoskeleton (incl legs)

A trace level of dimethylcyanamide revealed in the GCMS (see Figure 4) is the only direct evidence of cyanogeniccompounds in our sample

4 Discussion

The gomphodesmid millipedes utilized by the Bobo peopleare quite small (max 5 cm long) but gomphodesmids mayoccur in very large numbers at certain times of the yearThus Lewis [24] stated that 50 specimens of S sheribongensiscould be collected in 10ndash15 minutes at the beginning of therainy season in Zaria Nigeria and even (pers comm) thatit was sometimes impossible to take a step without crushingseveral individuals Barbetta et al [40] used 1200 specimensof Haplogomphodesmus pavani (Demange 1965) for theirbiochemical study an indication that also this species can bequite abundant Spirostreptids may occasionally form hugeswarms for example in Ghana and Zimbabwe [41] They are


Table 4 Summary of the current knowledge about utilization of millipedes by mammals as anointing andor food item

Mammals Millipedes Chemicals Use ReferenceMarsupialia

Opossum (Didelphis albiventris)

Leptodesmus dentellus(Chelodesmidae)

Gymnostreptus olivaceus(Spirostreptidae)

Benzoquinonesand Cyanogenics

Consumption andsniffing [47]

Carnivora

White-nosed coatis (Nasua narica) Orthoporus sp(Spirostreptidae) Benzoquinones

Consumption afterprey-rollingtreatment

[13]

Meerkat-mangoose (Suricata suricatta) Consumption aftertreatment [49 51]

Striped skunk (Mephitis mephitis) Consumption [52]Primates

Capuchin monkeys (Cebus sp) Orthoporus dorsovittatus(Spirostreptidae) Benzoquinones Self-anointing [19]

Capuchin monkeys (C olivaceus) Orthoporus dorsovittatus(Spirostreptidae) Benzoquinones Self-anointing [18]

Owl monkeys (Aotus sp) Anadenobolus monilicornis(Rhinocricidae) Benzoquinones Self-anointing [55]

Black lemurs (Elemur macaco) Charactopygus sp(Spirostreptidae) Benzoquinones Self-anointing [17]

Lemurs (Varecia rubra and Eulemur fulvus albifrons)

Self-anointing andconsumption after

handling andsalivating

[50 5372]

Humans Bobo population

Tymbodesmus falcatus andSphenodesmus sheribongensis

(Gomphodesmidae)unknown

Spirostreptidae


Consumption afterboiling and drying

Thisstudy

sometimes of considerable size up to 30 cm long Consid-ering the regular and high levels of some essential nutri-ents (Ca Fe PUFA) and seasonal abundance of millipedesthey likely represent a significant type of minilivestock forthe Bobo peopleThe ethnobiologicalethnopharmacologicaluses ofmany unconventional species that is earthworms andinsects are documented in several cultures being part ofa complex system of specific traditional knowledge adaptedto local resource availability and to very variable hygienicstandards [42 43] thatmay not fitwith the food-qualityfood-safety standards imposed in industrialized countries [44]

As mentioned above gomphodesmids belong to theldquocyanogenicrdquo millipedes [11] Cyanogenesis in millipedes andother arthropods has not been studied with state-of-artmethodology [41] but the presence of hydrogene cyanide andits precursors has been demonstrated in many species of theorder Polydesmida [11 26 27 45 46] including two species ofGomphodesmidae HCN has been identified in the secretionofAstrodesmus laxus (Gerstacker 1873) from East Africa [47]and its precursor mandelonitrile in Haplogomphodesmuspavani [40] Although there is no direct information onthe secretions of T falcatus and S sheribongensis there isno reason to believe that they have lost the cyanogenicfunction They have the same complement of defense glands

as the majority of gomphodesmids includingA laxus andHpavanii that is 11 pairs of glands opening along the sides ofthe body [23]

Benzoquinones have been detected in several spirostrep-tid species [11 28 48] In Figure 3 the defensive glands canbe seen as a series of darker spots one on each diplosegmentexcept the very first and last ones

Some birds and mammals (see Table 4) have beendescribed to use ldquoquinonerdquo millipedes for self-anointing inorder to control ectoparasites and mosquito biting rate andsome mammals like opossum coatis skunk mongoose andlemurs are known to consume toxic millipedesmdashmainlySpirostreptidaemdashafter different treatments such as prey-rolling handling and salivating [13 49ndash52] The complexbehaviours that precede ingestion require a considerableinvestment of time and energy and are evidently necessarymaybe in order to reduce toxicity of the millipedes to beeaten [53 54] The use of benzoquinonigenic plants for self-anointing in orangutans [54] and owl monkeys [55] supportsthe hypothesis that benzoquinones are used by primates fortheir specific biochemical defensive properties Contrarilyuse and consumption of gomphodesmid cyanogenic milli-pedes seems to be rare even in most omnivorous mammalsexcept opossum [56]


41 Cyanide in Traditional Foods The use of millipedes ashuman food is absolutely exceptional but the use of fooditems containing significant levels of cyanide is widespread[26] Over 2000 plant species contain cyanide as a defenseagainst insects and other herbivores [26] and the mostimportant cyanogenic crop is cassava (Manihot esculentaCrantz) a staple food of hundreds of millions of humans inthe tropics [57 58]

In the Amazonsmdashthe centre of domestication of cassa-vamdashthe more toxic bitter varieties named yuca amarga arethe most intensively cultivated because of their resistanceto pest insects and rodents However cassava domesticationlargely preceded malaria ingression in the New World [59]and the indigenous preparation is aimed at avoiding thecyanogenic compounds consisting of specific postharvestoperations grinding squeezing toasting and fermentation[58 60] Boiling alone is not enough to avoid the toxicityof yuca amarga [58] and only the sweet varieties of yucadulce which are low in glycosides can be consumed safely insoups or fried (MGP personal observation in Alto OrinocoVenezuela)

The Bobo people subboil the millipedes as part ofthe preparation for meals This treatment may degrade thecyanogenic compounds by releasing the hydrogen cyanidegas thereby detoxifying the gomphodesmids

Nonetheless the short (3ndash5 minutes) subboiling and nat-ural drying appear to be a specific treatment for millipedesmaybe aimed at preserving part of their chemicals Moreoverbenzoquinones are not readily soluble in water and themost characteristic juliform benzoquinone toluquinone isinsoluble in water [61] thus substantial amounts of themmayremain in the millipede bodies even after cooking

42 Biocultural Perspective African populations consumeraw and subboiled bitter cassava [58 60 62 63] as well asmany other bitter food items that would not be toleratedby other populations It has been proposed that the reducedsensitivity to bitterness is an ancient adaptation (70 kYBP)[64] of the human bitter-taste specific receptor to bitterantimalarial compounds (eg flavonoids) which are stillabundant in the West African diet [62 65] Cassava containsthe cyanogenic glucoside compounds linamarin and lotaus-tralin that once ingested are metabolized to thiocyanateand cyanate Notably thesemetabolites are biologically activealthough less toxic than cyanide and at levels of expecteddietary intake cyanide-related compounds (eg cyanate) areable to modify essential proteins of Plasmodium falciparumand inhibit parasite survival [66]

Considering that malaria represents the main cause ofmortality in the adult sub-Saharan population and that Burk-ina Faso is endemic for several Plasmodium species [67 68]biocultural adaptations aimed at controlling this pathogen arestrongly expected Natural benzoquinones are known to exertan antiplasmodic activity in vitro [69] and their metabolitesmight act as systemic repellents againstmosquitoes or as anti-malarial prophylaxis in the Bobos Moreover studies onWestAfrican populations have demonstrated strong links betweenmalaria sub-lethal cyanide intake from bitter cassava and

Figure 5 Scheme of the exceptional use of toxic millipedes asdemonstrated for capuchin monkeys that during the rainy seasonuse millipedesrsquo secretions against mosquitoes Bobo people inBurkina Faso may consume millipedes for their antiparasite effect

sickle-cell anemia [60 70] a genetic pathology affecting ery-throcytes that confers protection against Plasmodium Thusafter demonstrating that cyanide interacts with hemoglobinspartially compensating sickle-cell dysfunctionality [60 71]some authors proposed that the abundant consumption ofbitter foods could enhance biological fitness in West Africanpopulations exposed to malaria [62 70] Therefore bothethological and bioanthropological evidences suggest thatldquotoxicrdquo millipedes consumed by the Bobo people take part ina complex biocultural mechanism for malaria control

5 Concluding Remarks

Key topics of this paper are summarized in Figure 5Whethermillipedes will ever become a major actor in minilivestockhusbandry may be dubious but the Bobo people have shownthat they constitute a helpful food source for an ever-growinghuman population especially in rural Africa In addition thepotential ofmillipede chemicals for deterringmosquitoes andfor influencing Plasmodium and other parasites constitutes apromising field of research

Conflict of Interests

The authors declare that there is no conflict of interestsregarding the publication of this paper

Acknowledgments

The authors are grateful to the Bobo people at Kou forsharing their knowledge with them to The Fonds Franco-phone des Inforoutes (FFI) and the Cooperation Belge auDeveloppement for funding the 2011 expedition in Burk-ina Faso to Bakary Sanou for help with field work toDidier VandenSpiegel for identification of T falcatus andto Mika Zagrobelny for her attemptmdashagainst all oddsmdashto isolate cyanogenic compounds from alcohol-preservedmillipede specimensThe NMR spectrometers were acquiredin collaboration with the University of South Bohemia (CZ)with financial support from the European Union through


the EFRE INTERREG IV ETC-AT-CZ programme (ProjectM00146 ldquoRERI-uasbrdquo) Several people helped with discus-sion and suggestions in particular Natalie Vasey Jean ZidaRobert O Malley William C McGrew Linda C JacksonMarco Pombi Salvatore Musumeci and Paul Weldon

References

[1] FAO ldquoForest insects as food humans bite backrdquo in Proceedingsof a Workshop on Asia-Pacific Resources and their Potentialfor Development P B Durst D V Johnson R N Leslieand K Shono Eds Thailand Chiang Mai February 2008httpwwwfaoorgdocrep012i1380ei1380e00pdf

[2] N J Turner L J Luczaj P Migliorini et al ldquoEdible and tendedwild plants traditional ecological knowledge and agroecologyrdquoCritical Reviews in Plant Sciences vol 30 no 1-2 pp 198ndash2252011

[3] B A Rumpold and O K Schluter ldquoPotential and challenges ofinsects as an innovative source for food and feed productionrdquoInnovative Food Science and Emerging Technolologies vol 17 pp1ndash11 2013

[4] M G Paoletti T Gomiero and D Pimentel ldquoIntroductionto the special issue towards a more sustainable agriculturerdquoCritical Reviews in Plant Sciences vol 30 no 1-2 pp 2ndash5 2011

[5] M G Paoletti and S Bukkens ldquoMinilivestock sustainable useof biodiversity for human foodrdquo Ecology of Food and Nutritionvol 36 no 2ndash4 pp 95ndash346 1997

[6] M G Paoletti Ed Ecological Implications of MinilivestockPotential of Insects Rodents Frogs and Sails Science EnfieldNH USA 2005

[7] A van Huis ldquoInsects eaten in Africa (Coleoptera Hymeno-ptera Diptera Heteroptera Homoptera)rdquo in Ecological Implica-tions of Minilivestock M G Paoletti Ed pp 231ndash244 ScienceEnfield NH USA 2005

[8] DGA BOonincx J van ItterbeeckM JWHeetkampH vanden Brand J J A van Loon and A van Huis ldquoAn explorationon greenhouse gas and ammonia production by insect speciessuitable for animal or human consumptionrdquo PLoS ONE vol 5no 12 Article ID e14445 2010

[9] R W Pemberton ldquoContemporary use of insects and otherarthropods in traditional Korean medicine (Hanbang) in SouthKorea and elsewhererdquo inEcological Implications ofMinilivestockM G Paoletti Ed pp 459ndash474 Science Enfield NH USA2005

[10] M G Paoletti and D L Dufour ldquoEdible invertebrates amongAmazonian Indians a critical review of disappearing knowl-edgerdquo in Ecological Implications of Minilivestock M G PaolettiEd pp 293ndash342 Science Enfield NH USA 2005

[11] T Eisner D Alsop K Hicks and J Meinwald ldquoDefensive secre-tions of millipedesrdquo in Arthropod Venoms S Bettini Ed vol48 of Handbuch der Experimentellen Pharmakologie pp 41ndash72Springer Berlin Germany 1978

[12] S P Hopkin and H J Read The Biology of Millipedes OxfordScience 1992

[13] P J Weldon C F Cranmore and J A Chatfield ldquoPrey-rollingbehavior of coatis (Nasua spp) is elicited by benzoquinonesfrom millipedesrdquo Naturwissenschaften vol 93 no 1 pp 14ndash162006

[14] M Forthman and C Weirauch ldquoToxic associations a review ofthe predatory behaviors of millipede assassin bugs (Hemiptera

Reduviidae Ectrichodiinae)rdquo European Journal of Entomologyvol 109 pp 147ndash153 2012

[15] T Eisner M Eisner A B Attygalle M Deyrup and JMeinwald ldquoRendering the inedible edible circumvention ofa millipedersquos chemical defense by a predaceous beetle larva(Phengodidae)rdquo Proceedings of the National Academy of Sciencesof the United States of America vol 95 no 3 pp 1108ndash1113 1998

[16] J P Rood ldquoPopulation dynamics and food habits of the bandedmongooserdquo East AfricaWildlife Journal vol 13 no 2 pp 89ndash1111975

[17] C R Birkinshaw ldquoUse of millipedes by black lemurs to anointtheir bodiesrdquo Folia Primatologica vol 70 no 3 pp 170ndash1711999

[18] X Valderrama J G Robinson A B Attygalle and T EisnerldquoSeasonal anointment with millipedes in a wild primate achemical defense against insectsrdquo Journal of Chemical Ecologyvol 26 no 12 pp 2781ndash2790 2000

[19] P J Weldon J R Aldrich J A Klun J E Oliver and MDebboun ldquoBenzoquinones from millipedes deter mosquitoesand elicit self-anointing in capuchin monkeys (Cebus spp)rdquoNaturwissenschaften vol 90 no 7 pp 301ndash304 2003

[20] I Sazima ldquoAnting behaviour with millipedes by the dendroco-laptid birdXiphocolaptes albicollis in Southeastern Brazilrdquo BiotaNeotropica vol 9 no 1 pp 249ndash252 2009

[21] F Malaisse ldquoHuman consumption of Lepidoptera termitesorthoptera and ants in Africardquo in Ecological Implications ofMinilivestock M G Paoletti Ed pp 175ndash230 Science EnfieldNH USA 2005

[22] W A Shear ldquoClass diplopoda de blainville in gervais 1844rdquo inAnimal Biodiversity An Outline of Higher-Level Classificationand Survey of Taxonomic Richness Z Zhang -Q Ed vol 3148of Zootaxa pp 159ndash164 2011

[23] R L Hoffman Monograph of the Gomphodesmidae A familyof African PolydesmoidMillipedes Verlag des NaturhistorischenMuseums Wien Vienna Austria 2005

[24] J G E Lewis ldquoThe life history and ecology of the milli-pede Tymbodesmus falcatus (Polydesmida Gomphodesmida)in Northern Nigeria with notes on Sphenodesmus sheribongen-sisrdquo Journal of Zoology vol 164 pp 551ndash563 1971

[25] E Krabbe Systematik der Spirostreptidae vol 24 of Abhandlun-gen des Naturwissenschaftlichen Vereins in Hamburg (NF) 1982

[26] M Zagrobelny S Bak and B LMoslashller ldquoCyanogenesis in plantsand arthropodsrdquo Phytochemistry vol 69 no 7 pp 1457ndash14682008

[27] W A Shear T H Jones and H M Miras ldquoA possible phylo-genetic signal in milliped chemical defenses the polydesmidanmilliped Leonardesmus injucundus Shelley amp Shear secretesp-cresol and lacks a cyanogenic defense (Diplopoda Poly-desmida Nearctodesmidae)rdquo Biochemical Systematics and Ecol-ogy vol 35 no 12 pp 838ndash842 2007

[28] J Smolanoff J M Demange J Meinwald and T Eisner ldquo14-Benzoquinones in African millipedsrdquo Psyche vol 82 no 1 pp78ndash80 1975

[29] D E Reichle M H Shaks and D A Crossley ldquoCalciumpotassium and sodium content of forest floor arthropodsrdquoAnnals of the Entomological Society of America vol 62 no 1pp 57ndash62 1969

[30] C S Gist and D A Crossley Jr ldquoThe litter arthropod com-munity in a Southern Appalachian Hardwood Forest numbersbiomass and mineral element contentrdquo The American MidlandNaturalist Journal vol 93 no 1 pp 107ndash122 1975


[31] J Graveland and T van Gijzen ldquoArthropods and seeds are notsufficient as calcium sources for shell formation and skeletalgrowth in passerinesrdquo Ardea vol 82 no 2 pp 299ndash314 1994

[32] KNakamura and J Taira ldquoDistribution of elements in themilli-pede Oxidus gracilis C L Koch (Polydesmida Paradoxoso-matidae) and the relation to environmental habitatsrdquo Biometalsvol 18 no 6 pp 651ndash658 2005

[33] S Bures and K Weidinger ldquoSources and timing of calciumintake during reproduction in flycatchersrdquo Oecologia vol 137no 4 pp 634ndash641 2003

[34] R S Hames J D Lowe S B Swarthout and K V RosenbergldquoUnderstanding the risk to neotropical migrant bird species ofmultiple human-caused stressors elucidating processes behindthe patternsrdquo Ecology and Society vol 11 no 1 article 24 2006

[35] R Hood-Nowotny B Schwarzinger C Schwarzinger et alldquoAn analysis of diet quality how it controls fatty acid profilesisotope signatures and stoichiometry in the malaria mosquitoAnopheles arabiensisrdquo PLoS ONE vol 7 no 10 Article IDe45222 2012

[36] K J Kramer T L Hopkins and J Schaefer ldquoApplications ofsolids NMR to the analysis of insect sclerotized structuresrdquoInsect Biochemistry and Molecular Biology vol 25 no 10 pp1067ndash1080 1995

[37] A Collavo R H Glew H Yunk-Sheng et al ldquoHouse cricketsmall scale farmingrdquo in Ecological Implications of MinilivestockM G Paoletti Ed pp 519ndash544 Science Enfield 2005

[38] D Fontaneto M Tommaseo-Ponzetta C Galli P Rise R HGlew andM G Paoletti ldquoDifferences in fatty acid compositionbetween aquatic and terrestrial insects used as food in humannutritionrdquo Ecology of Food and Nutrition vol 50 no 4 pp 351ndash367 2011

[39] IOM Recommended Intakes for Individuals Elements Food andNutrition Board Institute of Medicine National AcademiesUSA 2004 httpwwwiomeduGlobalNews20Announce-mentssimmediaFilesActivity20FilesNutritionDRIsDRISummary Listingpdf

[40] M Barbetta G Casnati andM Pavan ldquoSulla presenza di D-(+)mandelonitrile nella secrezione difensiva del miriapode Gom-phodesmus pavani Demrdquo Memorie Della Societa EntomologicaItaliana vol 45 pp 169ndash176 1966

[41] J M Dangerfield and S R Telford ldquoAggregation in the tropicalmillipede Alloporus uncinatus (Diplopoda Spirostreptidae)rdquoJournal of Zoology vol 230 no 3 pp 503ndash511 1993

[42] G R DeFoliart F V Dunkel and D Gracer ldquoThe food insectsnewsletter chronicle of changing culturerdquo Aardvark Global2009

[43] E L CooperM Balamurugan Chih-YangHuang et al ldquoEarth-worms dilong ancient inexpensive noncontroversial modelsmay help clarify approaches to integrated medicine empha-sizing neuroimmune systemsrdquo Evidence-Based Complementaryand Alternative Medicine vol 2012 Article ID 164152 11 pages2012

[44] S Belluco C Losasso M Maggioletti C C Alonzi and M GPaoletti A Ricci ldquoEdible insects in a food safety and nutritionalperspective a critical reviewrdquo Comprehensive Reviews in FoodScience and Food Safety vol 12 no 3 pp 296ndash313 2013

[45] S E Makarov B P M Curcic V V Tesevic et al ldquoDefensivesecretions in three species of polydesmids (Diplopoda Poly-desmida Polydesmidae)rdquo Journal of Chemical Ecology vol 36no 9 pp 978ndash982 2010

[46] Y Kuwahara N Shimizu and T Tanabe ldquoRelease of hydrogencyanide via a post-secretion Schotten-Baumann reaction in

defensive fluids of polydesmoidmillipedesrdquo Journal of ChemicalEcology vol 37 no 3 pp 232ndash238 2011

[47] H E Eisner W F Wood and T Eisner ldquoHydrogen cyanideproduction in North American and African polydesmoid mil-lipedesrdquo Psyche vol 82 no 1 pp 20ndash23 1975

[48] R Deml and A Huth ldquoBenzoquinones and hydroquinones indefensive secretions of tropical millipedesrdquo Naturwissenscha-ften vol 87 no 2 pp 80ndash82 2000

[49] S P Doolan and D W Macdonald ldquoDiet and foraging behavi-our of group-livingmeerkats Suricata suricatta in the SouthernKalaharirdquo Journal of Zoology vol 239 no 4 pp 697ndash716 1996

[50] N Vasey ldquoCircadian rhythms in diet and habitat use in redruffed lemurs (Varecia rubra) and white-fronted brown lem-urs (Eulemur fulvus albifrons)rdquo American Journal of PhysicalAnthropology vol 124 no 4 pp 353ndash363 2004

[51] T Eisner and J A Davis ldquoMongoose throwing and smashingmillipedesrdquo Science vol 155 no 3762 pp 577ndash579 1967

[52] C N Slobodchikoff ldquoExperimental studies of tenebrionid bee-tle predation by skunksrdquo Behaviour vol 66 pp 313ndash322 1978

[53] N Vasey ldquoNiche separation inVarecia variegata rubra and Eule-mur fulvus albifrons II Intraspecific patternsrdquo American Jour-nal of Physical Anthropology vol 118 no 2 pp 169ndash183 2002

[54] H C Morrogh-Bernard ldquoFur-rubbing as a form of self-medi-cation inPongo pygmaeusrdquo International Journal of Primatologyvol 29 no 4 pp 1059ndash1064 2008

[55] M Zito S Evans and P J Weldon ldquoOwl monkeys (Aotus spp)self-anoint with plants andmillipedesrdquo Folia Primatologica vol74 no 3 pp 159ndash161 2003

[56] R T Santori ldquoDiscrimination of millipedes by the opossumDidelphis albiventris (Marsupiala Didelphidae)rdquo Journal of Ad-vanced Zoology vol 19 no 2 pp 118ndash119 1998

[57] FAORoots Tubers Plantains and Bananas inHumanNutritionFAO Rome Italy 1990 httpwwwfaoorgdocrept0207eT0-207E00htmContents

[58] W M Wilson and D L Dufour ldquoWhy ldquobitterrdquo cassava Pro-ductivity of ldquobitterrdquo and ldquosweetrdquo cassava in a Tukanoan Indiansettlement in theNorthwestAmazonrdquoEconomic Botany vol 56no 1 pp 49ndash57 2002

[59] R Carter and K N Mendis ldquoEvolutionary and historicalaspects of the burden ofmalariardquoClinicalMicrobiology Reviewsvol 15 no 4 pp 564ndash594 2002

[60] L C Jackson M Oseguera S Medrano and Y L Kim ldquoCar-bamylation of hemoglobin in vivo with chronic sublethal diet-ary cyanide implications for hemoglobin Srdquo Biochemical Medi-cine and Metabolic Biology vol 39 no 1 pp 64ndash68 1988

[61] V S Nithianandam K Kaleem F Chertok and S Erhan ldquoQui-none-amine polymers V Syntheses and solubilities of severaldiamine-p-benzoquinone oligomers (PAQ)rdquo Journal of AppliedPolymer Science vol 42 no 11 pp 2893ndash2897 1991

[62] L C Jackson ldquoEcological modelling of human-plant-parasitecoevolutionary triads heoretical perspectives on the interre-lationships of human HbPS GGPD Manihot esculenta Viciafaba andPlasmodium fakiparumrdquo inAdaptation toMalariaTheInteraction of Biology and Culture L Greene and M DanubioEds Academic New York NY USA 1996

[63] L C Jackson ldquoThe coevolutionary relationship of humans anddomesticated plantsrdquo American Journal of Physical Anthropol-ogy vol 101 no 23 pp 161ndash176 1996

[64] N Soranzo B Bufe P C Sabeti et al ldquoPositive selection on ahigh-sensitivity allele of the human bitter-taste receptor TAS2-R16 by the N172 allele may have driven the signal 8 of selection


at an early stage of human evolutionrdquo Current Biology vol 15no 14 pp 1257ndash1265 2005

[65] S S Maranz ldquoAn alternative paradigm for the role of anti-malarial plants in AfricardquoTheScientificWorld Journal vol 2012Article ID 978913 9 pages 2012

[66] R L Nagel C Raventos H B Tanowitz andMWittner ldquoEffectof sodium cynanate onPlasmodium falciparum in vitrordquo Journalof Parasitology vol 66 no 3 pp 483ndash487 1980

[67] P Rihet L Abel Y Traore T Traore-Leroux C Aucan and FFumoux ldquoHuman malaria segregation analysis of blood infe-ction levels in a suburban area and a rural area in Burkina FasordquoGenetic Epidemiology vol 15 no 5 pp 435ndash450 1998

[68] S-J Wang C Lengeler T A Smith et al ldquoRapid urban mala-ria appraisal (RUMA) I epidemiology of urban malaria inOuagadougourdquoMalaria Journal vol 4 article 43 2005

[69] P Grellier AMaroziene H Nivinskas J Sarlauskas A Alivertiand N Cenas ldquoAntiplasmodial activity of quinones roles ofaziridinyl substituents and the inhibition of Plasmodium falci-parum glutathione reductaserdquo Archives of Biochemistry andBiophysics vol 494 no 1 pp 32ndash39 2010

[70] L C Jackson ldquoTwo evolutionary models for the interactionsof dietary organic cyanogens hemoglobins and falciparummalariardquo American Journal of Human Biology vol 2 no 5 pp521ndash532 1990

[71] PNGillette JMManning andA Cerami ldquoIncreased survivalof sickle-cell erythrocytes after treatment in vitro with sodiumcyanaterdquo Proceedings of the National Academy of Sciences of theUnited States of America vol 68 no 11 pp 2791ndash2793 1971

[72] N Vasey ldquoNiche separation in Varecia variegata rubra andEulemur fulvus albifrons I Interspecific patternsrdquo AmericanJournal of Physical Anthropology vol 112 pp 411ndash431 2000


cyanide derived frommandelonitrile and related compounds(inmost flatbackedmillipedes superorderMerocheta)Thesetoxic smelly chemicals make millipedes unattractive formost predators although there are some animals vertebratesas well as invertebrates which eat millipedes [12 13] andsome are even specialized on a millipede diet for exampleassassin bugs (family Reduviidae) of the subfamily Ectri-chodiinae [14] and beetle larvae of the family Phengodidae[15] A few vertebrates are reported to eat millipedes asfor instance banded mongoose (Mungos mungo) [16] Somebirds and nonhominid primates use toxic millipedes forldquoself-anointmentrdquo presumably exploiting an insect-repellenteffect of the millipedesrsquo defensive chemicals especially ben-zoquinones [17ndash20]We have however not been able to traceany record of millipedes being used as food in any humansociety but nowwe can report the consumption ofmillipedesby the Bobo population of Burkina Faso a region whereentomophagy has been extensively described [21]

Themillipedeswhich are used as human food by the Bobobelong to two families Gomphodesmidae and Spirostrepti-dae

Gomphodesmidae (flatbacked millipedes of the orderPolydesmida) (Figures 1 and 2) belong to the ldquocyanogenicrdquomillipedes the family is endemic to the African continentsouth of the Sahara includes 146 named species [22] andwas monographed by Hoffman (2005) [23] The only gom-phodesmid species previously recorded from Burkina Fasois Tymbodesmus falcatus (Karsch 1881) collected in Oua-gadougou [23] One of the gomphodesmid species used forfood at Kou village in Burkina Faso is indeed T falcatus (DVandenspiegel det) the other is Sphenodesmus sheribongensis(Schioslashtz 1966) (HE det) T falcatus is known from MaliBurkina Faso Nigeria Sudan and Central African RepublicS sheribongensis was previously known from Ghana IvoryCoast andNigeria [23] Lewis [24] studied the life history andecology of both species in Zaria NorthernNigeria Both havea two-year life cycle Juvenile stadia of S sheribongensis liveentirely in the soil whereas adults can be extremely abundanton the soil surface during the first part of the rainy season(from May to July) T falcatus is similar except that the lastjuvenile (subadult) stadium is seasonally surface active likethe adults albeit in smaller numbers

Spirostreptidae (cylindrical millipedes of the order Spiro-streptida) (Figure 3) belong to the ldquoquinonerdquo millipedes thefamily is near-endemic to the Afrotropical and Neotropicalregions includes 275 named species [22] and was mono-graphed by Krabbe [25] No species of Spirostreptidae havebeen recorded from Burkina Faso and the one occurring atKou has not yet been identified

We do not have any strong evidence that T falcatus andS sheribongensis actually produce hydrogen cyanide nor thatthe spirostreptid in question produces quinones The generaloccurrence of these substances in the respective higher taxato which the species belong however is strong circumstantialevidence that this is actually so although recent studieshave demonstrated a larger diversity in millipede defensivechemicals than previously assumed [26ndash28]

Until this study the nutritional value of millipedes hasnever been assessed Considering that the muscle volume

Figure 1 Live gomphodesmid millipede (Tymbodesmus falcatus)from Kou S Tchibozo phot 2011

Figure 2 gomphodesmid millipede from Kou after boiling S Tchi-bozo phot 2012

Figure 3 Spirostreptidmillipede fromKou S Tchibozo phot 2012

of millipede is small they will constitute a poor source ofproteinTheir gutsmainly contain soil and litter remains [24]but their calcified exoskeletonmight constitute a considerablesource of calcium which may constitute 13ndash17 of the dryweight [29ndash31] or 9 of the fresh weight [32] In factmillipedes are considered an essential source of calcium foregg production in certain birds [30 33 34] In this workwe present data based on Tymbodesmus falcatus one of thespecies eaten by the Bobo people

2 Material and Methods

21 Sampling and Ethnobiological Data Collection Obser-vations and interviews with the Bobo people were made






3)2for calib-

ration [36]






3 Results





No ID Name 119905119877




0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18Time (min)

0102030405060708090

100 737906

873

582 17153

4 7

10

9

5

1214

NN

Rela

tive a

bund

ance


119877


737) (5) and oleic acid 16 0 (119905119877






4 Discussion










Carnivora



[13]










[50 5372]




Spirostreptidae



Thisstudy



















Acknowledgments




References



















































































3)2for calib-

ration [36]






3 Results





No ID Name 119905119877




0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18Time (min)

0102030405060708090

100 737906

873

582 17153

4 7

10

9

5

1214

NN

Rela

tive a

bund

ance


119877


737) (5) and oleic acid 16 0 (119905119877






4 Discussion










Carnivora



[13]










[50 5372]




Spirostreptidae



Thisstudy



















Acknowledgments




References
















































































No ID Name 119905119877




0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18Time (min)

0102030405060708090

100 737906

873

582 17153

4 7

10

9

5

1214

NN

Rela

tive a

bund

ance


119877


737) (5) and oleic acid 16 0 (119905119877






4 Discussion










Carnivora



[13]










[50 5372]




Spirostreptidae



Thisstudy



















Acknowledgments




References






















































































Carnivora



[13]










[50 5372]




Spirostreptidae



Thisstudy



















Acknowledgments




References



























































































Acknowledgments




References
















































































References














































































Millipedes as Food for Humans: Their Nutritional and Possible Antimalarial Value—A First Report

Documents

Transcript of Millipedes as Food for Humans: Their Nutritional and Possible Antimalarial Value—A First Report