Fleas and flea-borne diseases of North Africa

Fleas and lea-bo ne diseases o No h A ica. 1 Basma El Hamzaoui1,2; An onio Zu i a3*; C is ina Cu illas3, Philippe Pa ola1,2 2 1Aix Ma seille Uni ; IRD; AP-HM; SSA; VITROME; 3 2IHU Médi e anée In ec ion; Ma seille; F ance 4 3Depa men o Mic obiology and Pa asi ology. Facul y o Pha macy. Uni e si y o Se ille. 5 P o eso Ga cía González 2, 41012 Se ille, Spain. 6 Emails: BE: [email p o ec ed]; AZ: [email p o ec ed]; CC: cu ill[email p o ec ed]; PP : 7 [email p o ec ed] 8 *Co esponding au ho : D . An onio Zu i a. 3Depa men o Mic obiology and Pa asi ology. 9 Facul y o Pha macy. Uni e si y o Se ille. P o eso Ga cía González 2, 41012 Se ille, Spain. 10 Email add ess: [email p o ec ed] 11 12 13 14 15 16 17 18 19 20 Con en 21 Abs ac 22 In oduc ion 23 Fleas o medical- e e ina y impo ance in No h A ica 24  Pulex i i ans 25  Echidnophaga gallinacea 26  C enocephalides elis 27  C enocephalides canis 28  Spilopsyllus cuniculi 29  Xenopsylla cheopis 30  A chaeopsylla e inacei 31 Flea-bo ne diseases o No h A ica 32  Plague 33  Flea-bo ne icke sial diseases 34  Mu ine yphus 35  Ricke sia elis in ec ion and lea-bo ne spo ed e e 36  Ba onelloses 37  Ca -sc a ch disease 38  T ench e e 39  Pa asi ic diseases 40  Dipylidium caninum in ec ion o Dipylidiasis 41  Flea-bo ne i al diseases 42 Means o lea con ol and iden i ica ion 43 44 Abs ac 45 No h A ica has an in e es ing and ich wildli e including hema ophagous a h opods, and 46 speci ically leas, which cons i u e a la ge pa o he No h A ican auna, and a e ecognised 47 ec o s o se e al zoono ic bac e ia. Flea-bo ne o ganisms a e widely dis ibu ed h oughou 48 he wo ld in endemic disease oci, whe e componen s o he enzoo ic cycle a e p esen . 49 Fu he mo e, lea-bo ne diseases could e-eme ge in epidemic o m because o changes in he 50 ec o –hos ecology due o en i onmen al and human beha iou modi ica ions. We need o 51 know he eal incidences o lea-bo ne diseases in he wo ld due o his incidence could be much 52 g ea e han a e gene ally ecognized by physicians and heal h au ho i ies. As a esul , diagnosis 53 and ea men a e o en delayed by heal h ca e p o essionals who a e unawa e o he p esence 54 o hese in ec ions and hus do no ake hem in o conside a ion when a emp ing o de e mine 55 he cause o a pa ien ’s illness. In his con ex , his bibliog aphic e iew aims o summa ise he 56 main species o leas p esen in No h A ica, hei geog aphical dis ibu ion, lea-bo ne 57 diseases, and hei possible e-eme gence. 58 59 Keywo ds: Fleas, No h A ica, lea-bo ne diseases, ec o s 60 61 Lis o abb e ia ions: 62 MALDI–TOF MS: Ma ix Assis ed Lase Deso p ion Ionisa ion – Time O Fligh Mass 63 Spec ome y 64 In oduc ion 65 Fleas (Insec a, Siphonap e a) a e obliga e hema ophagous ec opa asi es. They a e wingless 66 insec s which a e small (2-10 mm) and gene ally p esen a la e ally la ened body. Fleas ha e 67 h ee ho acic segmen s, each wi h a pai o well-de eloped legs (Beaucou nu and Launay, 68 1990; Bi am e al., 2010) enabling adul s o jump long dis ances (Bi am e al., 2010). They a e 69 ec opa asi es ha usually in es mammals and oden s bu a ely bi ds. Flea species dis ibu ion 70 ex ends o he se en con inen s, including An a c ica (Whi ing e al., 2008). Fu he mo e, leas 71 a e able o inhabi a e y wide ange o habi a s and hos s (Whi ing, 2002). The g ea es 72 di e si y o species can be obse ed in he empe a e egions o he globe (Lewis, 1993). 73 Fleas a e holome abolous insec s ha comple e hei cycle om egg o adul ia h ee la al 74 s ages and a pupal s age (Zakson-Aiken e al., 1996). The comple ion o he en i e li e cycle 75 a ies among species bu akes on a e age h ee o i e weeks depending on he empe a u e 76 and humidi y condi ions. Flea la ae a e e mi o m and legless, wi h chewing mou hpa s 77 (D yden and Rus , 1994). 78 Flea species adap o hei hos s bu do no do so exclusi ely. This ac could explain he ole 79 o he so-called a lea Xenopsylla cheopis (Ro hschild, 1903) in he epidemiology o human 80 plague, o he exis ence o highly p omiscuous leas species, such as he so-called human lea, 81 Pulex i i ans (Linnaeus, 1758) o he ca lea C enocephalides elis (Bouché, 1835) which 82 occu on a wide a ie y o ca ni o ous animal species (G a z, 1999). 83 Fleas’ beha iou owa ds hei hos s makes i possible o classi y hem in h ee ca ego ies, 84 including i) leas ha li e pe manen ly on hei hos , such as X. cheopis, P. i i ans, 85 C enocephalides canis and C. elis, ii) leas ha pe manen ly li e in hei hos s’ nes o bu ows, 86 pa asi izing hem only du ing blood meals, such as Ce a ophyllus gallinae; iii) and he so-called 87 pene a ing leas, such as emales o he gene a Tunga and Neo unga, which a e able o bu ow 88 in o hei hos s’ de mal issue, whe e hey inc ease d ama ically in size (up o l000- old) 89 accompanied by an ex ensi e mo phological degene a ion (Du den and T aub, 2002). Females 90 o Echidnophaga gallinacea can also ix hemsel es a ound he eyes o poul y a e 91 e iliza ion (F anc, 1994a). 92 Wi hin Siphonap e a, he Pulicidae amily exhibi s an in e es ing di e si y o hos speci ici y 93 pa e ns and ecological habi s (Beu el e al., 2008). Wi hin his amily, P. i i ans, C. elis and 94 e en X. cheopis ha e been he mos s udied species due o hei cosmopoli an dis ibu ion 95 oge he wi h he ac ha hese species a e closely ela ed o humans (Du den and T aub, 2002). 96 The impo ance o leas in human public heal h is mos ly ela ed o hei abili y o ansmi 97 in ec ious disease agen s du ing he blood meal. Some leas a e indeed ec o s o human 98 in ec ious diseases, such as bubonic plague, caused by Ye sinia pes is (Zeppelini e al., 2016), 99 mu ine yphus, caused by Ricke sia yphi (Peniche La a e al., 2012) and lea-bo ne spo ed 100 e e caused by Ricke sia elis (Angelakis e al., 2016) . They a e also p obably in ol ed in 101 he ansmission o Ba onella henselae he agen o ca -sc a ch disease (Bi am e al., 2010; 102 Chomel and Kas en, 2010). 103 “No h A ica” is a collec i e e m including Medi e anean coun ies and e i o ies si ua ed 104 in he no he n-mos egion o he A ican con inen , including Mo occo, Alge ia, Tunisia, 105 Libya and Egyp . I is a o al a ea o a ound i e million kilome es, mo e han 90% o which 106 is dese (Peel e al., 2007). No h A ica is ulne able o a ious clima es, wi h he coas being 107 cha ac e ised by a Medi e anean clima e o we win e s and d y summe s, while non-coas al 108 a eas a e cha ac e ised by an a id dese clima e wi h gene ally ho summe s, cold win e s and 109 li le ain all (Radhouane, 2013). This clima ic di e si ica ion con ibu es o he ichness o he 110 No h A ican auna, including leas. 111 In his e iew, we ocus on leas o e e ina y and clinical impo ance om No h A ica, 112 pa icula ly hose species belonging o he Pulicidae amily. We also discuss inno a i e 113 me hods o he iden i ica ion o leas and hei associa ed pa hogens. 114 Fleas o medical- e e ina y impo ance in No h A ica 115 The Pulicidae amily consis s o ou ibes, 21 gene a and 167 species (Beu el e al., 2008). 116 Some au ho s (Lewis, 1998) conside ed Pulicidae as including Tungidae. Howe e , Whi ing e 117 al. (2008) placed his amily as a monophyle ic and phylogene ically dis an g oup om 118 Tungidae (Beu el e al., 2008). Mos leas o e e ina y impo ance a e g ouped in his amily 119 since hey may ac as ec o s o some in ec ious diseases, may play a ole as in e media e hos s 120 o se e al pa asi es, and many cause alle gic eac ions in animals and humans, associa ed wi h 121 hei bloodsucking habi s (Doble and P e e , 2011). 122  Pulex i i ans (Figu e 1.1): 123 This lea has a cosmopoli an dis ibu ion and is o en e e ed as he “human lea”. Howe e , 124 his species o en pa asi izes a wide a ie y o hos s, including la ge wild mammals and oden s, 125 al hough a ely ound on a s (G a z, 1999). Unde na u al condi ions, his lea is ac i e 126 h oughou he yea , wi h a peak in summe (Beaucou nu and Launay, 1990). I s legs a e 127 su icien ly de eloped o jump up o 50 cm and i has he pa icula i y o being able o su i e 128 o a yea wi hou eeding, since his lea is capable o abso bing quan i ies o blood equi alen 129 o 20 imes i s weigh (Bel ho e al., 2015). 130 The p esence o his lea in No h A ica has been epo ed se e al imes. P. i i ans was 131 collec ed on dogs om Tunisia (Tunis, Mak a , Siliana). P.i i ans was also ound in no h- 132 wes e n Libya (T ipoli) on eigh a m dogs (Kaal e al., 2006), and in Mo occo (Agadi , 133 Casablanca,Tizni ) (Boudebouch e al., 2011). G. Blanc and M. Bal aza d (1945) long suspec ed 134 he ole o P. i i ans in he ansmission o he plague in Mo occo (Audouin-Rouzeau, 2003). 135 They conduc ed nine expe imen s wi h na u ally in ec ed P. i i ans on plague ic ims. In he 136 i s eigh expe imen s, all guinea pigs we e no in ec ed, e en when 240 leas we e used. Fo 137 hei nin h expe imen , G. Blanc and M. Bal aza d (1945) collec ed 720 leas om six plague 138 ic ims, and one guinea pig bi en by a lea was in ec ed. This expe imen enabled he au ho s 139 o s a e ha in ec ed human leas could ansmi diseases (Audouin-Rouzeau, 2003). 140  Echidnophaga gallinacea (Figu e 1.2): 141 This species is commonly known as he “S ick igh lea” and is abou 2 mm long. Females o 142 his species a e able o emain a ached o up o six weeks o a single hos si e, causing 143 ulce a ion a he a achmen si e. The eggs a e hen deposi ed in he ulce s ha a e o med on 144 he skin o he hos . La e , he la ae all o he g ound and eed on any o ganic deb is ound 145 (Bough on e al., 2006). In many cases, a la ge numbe o leas can cong ega e a ound he eyes 146 and on he ba e skin o poul y, which makes i di icul o emo e hem (Els on, 2001). 147 E. gallinacea is mo e ac i e in summe , causing se ious ouble wi h li es ock, especially in 148 u al a eas. In No h A ica, E. gallinacea has been epo ed in Mo occo (Raba , Agadi , Salé, 149 Sa i, Essaoui a, Casablanca), Alge ia (Saïda), in Tunisian islands (Dje ba and Zemb a) 150 (Beaucou nu and Launay, 1990) and has also been isola ed om dogs in Libya (Kaal e al., 151 2006). 152 Lo is e al. (2006) conduc ed a su eillance s udy o leas on mammals and associa ed 153 pa hogens in Egyp , wi h E. gallinacea being he main species collec ed. In addi ion, hese 154 au ho s we e able o de ec spo ed e e by Ricke sia sp, simila o RF2125, in all collec ed 155 specimens (Lo is e al., 2006). 156  C enocephalides elis (Figu e 1.3): 157 C. elis is commonly known as he “ca lea”. C. elis o igina es om A ica and now pa asi es 158 pe s and li es ock e y easily and egula ly in wa m and ho clima es (Beaucou nu and Ménie , 159 1998). I is conside ed o be he main lea species in ec ing domes ic ca ni o es in many 160 coun ies a ound he wo ld (Rus , 2016). 161 This species is cosmopoli an and seden a y on i s hos and is mainly ac i e in summe . All 162 mammals li ing in he same bio ope a e suscep ible o be in es ed by C. elis (Beaucou nu and 163 Launay, 1990). I s hos speci ici y is low, hus, i can be ound on o he animals, including small 164 uminan s, ca le, p ima es, oden s, poul y and e en opossums. The di ec ansmission 165 be ween indi iduals is equen , al hough i appea s o be a e in dogs han in ca s (Doble and 166 P e e , 2011). 167 C. elis has been ound in Tunisia, and has been collec ed om ca s, dogs, sheep and goa s. 168 Se e al mic oo ganisms ha e been de ec ed in C. elis in Tunisia, including Ba onella spp., R. 169 elis (Zoua i e al., 2017). I has also been collec ed om hedgehogs in Alge ia. These au ho s 170 epo ed he p esence o R. elis DNA in all C. elis collec ed. The esul s o his s udy can 171 he e o e help human and e e ina y clinicians o ocus on a b oade spec um o pa hogens and 172 ake hem in o conside a ion du ing diagnosis (Leulmi e al., 2016). 173 The p esence o his lea has also been epo ed in Mo occo (Casablanca, Tizni ) on domes ic 174 animals, and molecula biology demons a ed he p esence o Ba onella cla idgeiae, B. 175 henselae and R. elis in hese leas (Boudebouch e al., 2011). I has also been collec ed om 176 ca s and dogs om a a m in Libya (Kaal e al., 2006). A his poin , i should be highligh ed 177 he ausence o Ricke sia asembonensis in C. elis collec ed om No h A ica. This pa hogen 178 has been widely de ec ed in C. elis and C. canis collec ed om se e al egions o sub‐Saha an 179 A ica such as Rwanda, Zambia o Kenya (Nziza e al., 2018; Moonga e al., 2019), howe e ; 180 cu en ly, he e is no s udies which con i m he p esence o his bac e iim in he ca lea om 181 No h A ica. 182 This lea can quickly change hos s, which can play a ole in he ansmission o pa hogens. The 183 sali a o C. elis has i i a ing p ope ies, which gene ally leads o de ma osis due o he bi es. 184 I can also cause anemia du ing a massi e in es a ion (Gague e and P elaud, 2006). 185  C enocephalides canis (Figu e 1.4): 186 C. canis is also known as he “dog lea”. Despi e i s name, i has been demons a ed ha he 187 p e alence o C. elis in dogs is highe han ha o C. canis (Lina di and San os, 2012). C. canis 188 has a e y simila mo phology o C. elis subspecies. Thus, we can easily disc imina e be ween 189 C. canis and C. elis elis bu i used o be di icul o di e en ia e be ween C. canis and C. elis 190 s ongylus o C. elis o ien is. These di e ences a e mainly based on he shape o he ons o 191 he cephalic capsule, he p esence and shape o he do sal inc assa ion and he numbe o se ae 192 on he occipu (Lina di and San os, 2012). They also di e en ia e in he a e age jump heigh , 193 which is 15.5 cm in C. canis and 13.2 cm in C. elis (Beaucou nu and Launay, 1990). Al hough 194 bo h species a e ecognised ec o s o R. elis and se e al Ba onella spp. pa hogens, i is 195 impo an o disc imina e be ween C. elis and C. canis, since many au ho s ha e epo ed a 196 much lowe p e alence o hese bac e ia in C. canis han in C. elis (Kumsa e al., 2014; 197 Law ence e al., 2015). 198 This is a species ha is seden a y on i s hos and in es s mainly domes ic and wild canids. The 199 ed ox is i s p ima y hos , bu epidemiological s udies ha e shown ha C. canis can also be 200 ound on ca s, albei wi h a lowe p e alence han C. elis (Beaucou nu and Ménie , 1998; 201 Lina di and San os, 2012; Ma ugal e al., 2013). 202 This species o lea is equen in No h A ica, and has been collec ed om dogs om Tunisia, 203 whe e molecula analysis showed he p esence o Ba onella spp. in 23.5% o collec ed C. canis 204 (Zoua i e al., 2017). A p elimina y s udy conduc ed in Egyp on domes ic oden s e ealed he 205 p esence o C. canis in he Dakahlia go e no a e (Soliman and Mikhail, 2011). In no he n 206 amily Ricke siaceae (Maina e al., 2016). R. elis, R. asembonensis, and “Candida us 349 Ricke sia senegalensis” belong o he spo ed e e g oup icke siae (SFGR) ha gene ically 350 clus e s wi hin he ansi ional g oup o icke siae (Gillespie e al., 2007). We ha e jus 351 men ioned ha R. elis is associa ed wi h lea-bo ne spo ed e e , howe e , he pa hogenici y 352 o R. asembonensis and “Ca. R. senegalensis” is cu en ly unknown. These h ee agen s ha e a 353 wo ldwide dis ibu ion and hey ha e been de ec ed in humans and non-human p ima es (Tay 354 e al., 2015; Kho e al., 2016). R. asembonensis DNA has been de ec ed in leas om h ee 355 amilies o leas (Pulicidae, Ce a ophyllidae and Cop opsyllidae) wi h highes p e alence a es 356 epo ed in C. elis and C.canis (Rouche e al., 2012; Jiang e al., 2013). In spi e o ha , in 357 No h A ica his bac e ium only has been de ec ed in E. gallinacea collec ed om Egyp 358 (Lo is e al., 2006). 359 Ba onelloses 360 Ba onelloses a e zoonosis caused by G am-nega i e ae obic bac e ia o he genus Ba onella 361 (Iannino e al., 2018). Many species ha e been desc ibed. They include ecognized pa hogens 362 as well as bac e ia o unknown pa hogenici y. (S uckey e al., 2017). 363  Ca -sc a ch disease 364 This disease was i s desc ibed in 1889 and i is caused by B. henselae (Wong e al., 1995). 365 The domes ic ca is he main ese oi o his bac e ium (Chomel, 1996). 366 T ansmission o humans akes place h ough a ca sc a ch o bi e, bu con amina ion by C. elis 367 lea aeces and ansmission by egu gi a ion ha e also been demons a ed (Bouhsi a e al., 368 2013b). In ec ion wi h B. henselae can cause e e , hepa i is, endoca di is, bacilla y 369 angioma osis and bacilla y peliosis (Du den and T aub, 2002). 370 B. henselae has been de ec ed by qPCR in dog blood om Algie s, Alge ia (Azzag e al., 2012; 371 Bessas e al., 2016). B. henselae has also been de ec ed in C. canis and C. elis, collec ed om 372 domes ic animals in S ax, Jendouba and Manouba in Tunisia, (Belkhi ia e al., 2017; Zoua i e 373 al., 2017). This bac e ia was de ec ed using qPCR in C. elis collec ed om sheep, ca s and dogs 374 in Casablanca, Mo occo (Boudebouch e al., 2011). 375  T ench e e : 376 T ench e e is a zoonosis caused by in ec ion by he bac e ium B. quin ana, a g am-nega i e 377 bac e ium which is conside ed as a e-eme ging human pa hogen (Ande son and Neuman, 378 1997; Faccini-Ma ínez e al., 2017). This disease was desc ibed du ing he Second Wo ld Wa 379 ollowing se e al cases o soldie s who su e ed om he disease (Kos zewski, 1949). B. 380 quin ana may also be esponsible o bacilla y angioma osis (Relman e al., 1990), endoca di is 381 (D ancou e al., 1995) and ch onic lymphadenopa hy (Raoul e al., 1994). 382 I is ansmi ed by body lice (P. h. humanus) (Coulaud e al., 2014), bu some au ho s ha e 383 ecen ly shown he abili y o C. elis o ansmi B. quin ana, which was ound in lea aeces 11 384 days a e an in ec ious meal (Bouhsi a e al., 2013a; Ke ni e al., 2014). This bac e ium has 385 also been de ec ed in P. i i ans in Gabon (Rolain e al., 2005). 386 The p esence o his bac e ium has o en been de ec ed by molecula biology in body lice 387 collec ed om homeless people in no he n Alge ia (Louni e al., 2018) and also in pa ien s 388 du ing an endoca di is s udy conduc ed in Casablanca and Ma akech (Mo occo) (Boudebouch 389 e al., 2017), and, inally, in 12 endoca di is pa ien s in S ax (Tunisia) (Znazen e al., 2005). 390 Pa asi ic diseases 391  Dipylidium caninum in ec ion o Dipylidiasis 392 This is a medium-sized apewo m which equen ly pa asi izes he small in es ine o dogs and 393 ca s. Ne e heless, hey can occasionally pa asi ize humans (Mosk ina and E molenko, 2016), 394 in which case i causes Dipylidiasis. I s in e media e hos s a e C. elis and C. canis and i s inal 395 hos s a e dogs and ca s. These pa asi es deple e he hos s’ nu ien s du ing hei p esence in he 396 diges i e ac and he spolia ion o all he nu ien s explains he clinical signs (Nei a O e al., 397 2008). The pa asi e is ansmi ed o he hos by inges ion o con amina ed leas o aeces. 398 Fu he mo e, lice can, excep ionally, ansmi he wo m (F anc, 1994b). 399 This pa asi e has been epo ed in h ee coun ies o no he n A ica. I I was men ioned in a 400 s udy ha a ge ed wild canids in Tunisia, when he pa asi e was de ec ed in 55% o oxes and 401 jackals (Lahma e al., 2014). I was also de ec ed in Egyp , in a s udy o in es inal pa asi es 402 om 113 samples o s ay ca aeces aken no h o he Nile del a (Khala alla, 2011). E en in 403 Mo occo, D. caninum has been ound in s ay dogs in u ban and u al a eas o Raba wi h a 404 pe cen age o 40.4% o all samples es ed (57 dogs) (Pandey e al., 1987). 405 Flea-bo ne i al diseases 406 The biological ansmission o i uses by leas has no been widely s udied, he only known 407 i us ansmi ed by leas is Myxoma i us (myxoma osis i us) (Sobey e al., 1977)(Ke e al., 408 2015). This i us is ansmi ed by he abbi lea, S. cuniculi. This in ol es mechanical 409 con amina ion by mou h pa s and he i us is eleased when a bi e occu s (Shephe d and 410 Edmonds, 1980). Clinical signs a e usually se e e and dea h occu s wi hin 10 o 12 days. 411 Howe e , abbi s wi h milde signs, including hose su e ing om he amyxoma ous o m o 412 hose ha ha e been p e iously accina ed, may su i e wi h nu sing ca e (Me edi h, 413 2013).The epidemiology o his i us in No h A ica has ne e been s udied, bu he p esence 414 o he lea ec o has been epo ed in se e al Mo occan ci ies (Beaucou nu and Launay, 1990). 415 Al hough se e al human i al pa hogens ha e been isola ed o de ec ed in leas, he ole o leas 416 in hei ansmission is ei he unknown o conside ed o be inciden al. These i uses include 417 hose causing lymphocy ic cho iomeningi is, ick-bo ne encephali is and Russian sp ing- 418 summe encephali is (Du den and T aub, 2002). 419 The eline leukaemia i us (FeLV) is a equen i us in domes ic ca s and i was s ill suspec ed 420 o be ansmi ed by an a h opod ec o . Some au ho s (Vobis e al. 2003) de eloped an 421 expe imen al model o s udy he ec o capaci y o C. elis o ansmi he FeLV i us (Vobis 422 e al., 2003). The leas we e ed o 24 hou s wi h blood om a FeLV-in ec ed ca , and FeLV 423 was inally de ec ed in leas and hei aeces. The leas could e en ansmi he FeLV i us om 424 one blood sample o ano he . The esul s indica e ha ca leas a e po en ial ec o s o FeLV 425 RNA in i o and p obably also in i o (Vobis e al., 2003) 426 Iden i ica ion and labo a o y ea ing 427 The co ec iden i ica ion o lea species is essen ial in any esea ch o con ol p ojec . Cu en 428 iden i ica ion me hods a e mainly based on mo phological iden i ica ion. Howe e , o pe o m 429 de ailed mo phological iden i ica ion, i is necessa y o clea lea samples wi h 10% KOH o 430 NaOH (Lewis, 1993), examine unde a s e eomic oscope, and hen moun and pho og aph hem. 431 Molecula biology has been used o e he las 20 yea s o lea iden i ica ion and de ec ion o 432 hei associa ed pa hogens (Zu i a e al., 2015). These app oaches a e limi ed by he leng h o 433 ime, he a ailabili y o e e ence sequences in he GenBank da abase, he cos associa ed wi h 434 molecula biology app oaches, as well as he small numbe o en omologis s specialised in lea 435 axonomy (Yssou e al., 2016). Recen ly, mass spec ome y has eme ged as an inno a i e 436 iden i ica ion ool o a h opods, especially leas (Yssou e al., 2014). The use o MALDI TOF 437 MS equi es he de elopmen o an adequa e p o ocol in o de o s anda dise sample p epa a ion 438 me hods and o allow o subsequen exchanges using he da abase be ween se e al esea ch 439 labo a o ies (Nebbak e al., 2017). In addi ion, MALDI TOF MS has p o ed i s e ec i eness 440 in iden i ying leas which may o may no be in ec ed by a pa hogen and e en in dis inguishing 441 be ween leas in ec ed wi h wo pa hogens o he same amily (El Hamzaoui e al., 2018). 442 Ne e heless, since MALDI-TOF MS echniques ha e demons a ed some di e ences in he 443 MS spec a o specimens p ese ed in di e en s o age condi ions (Nebbak e al., 2017; Zu i a 444 e al., 2018), i is s ill necessa y o combine mo phological, molecula and p o eomics me hods 445 in o de o ca y ou an e icien speci ic iden i ica ion wi hin he O de Siphonap e a. 446 Flea ea ing is also a key s ep in he s udy o he biology o Siphonap e a, hei mo phology and 447 hei ec o ial capaci y. The ca lea, C. elis, is ound wo ldwide and has been epo ed o 448 pa asi ize many species o wild and domes ic animals (Rus and D yden, 1997). In addi ion, C. 449 elis has been desc ibed as ha ing low hos speci ici y. Indeed, i is a lea ha eeds on a a ie y 450 o animals and oden s, and is he e o e he igh choice o de eloping a labo a o y b eeding 451 p og amme (D yden and Rus , 1994). The a i icial ea ing sys em desc ibed by Wade and 452 Geo gi (1988) includes a hea ed Plexiglas box om which lea chambe s a e suspended and in 453 which a sou ce o human blood is hea ed (p e iously s o ed a 4°C) (Wade and Geo gi, 1988). 454 To main ain he equi ed empe a u e di e ence o 10°C, he en i e sys em is housed in a 455 empe a u e-con olled chambe . The empe a u e should be main ained be ween 25°C and 456 35°C, wi h a ela i e humidi y o 75 o 80% using a ay illed wi h wa e . Fleas a e aised in 457 he da k 24 hou s a day (Ke ni e al., 2015). 458 Rea ing leas makes i possible o s udy hei ec o ial capaci y. An expe imen al model o 459 a i icial in ec ion o leas wi h a s ain o B. quin ana has shown he abili y o C. elis o acqui e 460 he bac e ium and ansmi i , ali e, in aeces. Se e al lea g oups we e ed wi h blood mixed 461 wi h he bac e ial inoculum a di e en concen a ions. qPCR showed he p esence o B. 462 quin ana in aeces and immunohis ochemis y localised he bac e ium in he diges i e ac o 463 C. elis (Ke ni e al., 2014). 464 Con ol 465 Insec icides a e he mos widely-used means o lea con ol, including powde sp ays in nes s, 466 bu ows, o house walls in in es ed a eas (Rus , 2016). Mol inhibi o s a e also used o con ol 467 leas in hei la al s ages. I is s ongly ecommended o con ol hei hos in combina ion wi h 468 lea con ol using insec icides, in o de o a oid wha happened in he case o plague (F anc, 469 1994a). 470 Conclusion: 471 Fo se e al decades, we ha e wi nessed he e-eme gence o se e al ec o -bo ne zoono ic 472 pa hologies. Me agenomics and molecula biology ha e e olu ionized he epidemiology o 473 hese diseases; howe e , some a eas emain poo ly explo ed as No h A ica. 474 Fleas a e hema ophagous, wingless insec s ha ha e he abili y o jump. Thei abili y o ansmi 475 pa hogens explains hei impo ance in human and animal heal h. They a e sp ead all o e he 476 wo ld and some species do no equi e he p esence o a speci ic hos . This e iew summa izes 477 he la es da a on lea ec o s and lea-bo ne diseases in No h A ica (Mo occo, Alge ia, 478 Tunisia, Egyp and Libya). 479 Fo his eason, we ha e selec ed disease’s ec o species desc ibed in No h A ica in ecen 480 yea s, we ha e also epo ed he cases o ec o -bo ne diseases by leas diagnosed o upda e he 481 epidemiological si ua ion in his egion. 482 483 Decla a ions: 484 All au ho s consen o he publica ion 485 E hics app o al and consen o pa icipa e: no applicable 486 No compe ing in e es s exis 487 Funding: no applicable 488 A ailabili y o da a and ma e ials: no applicable 489 Au ho Con ibu ions: 490 W o e he pape : BE AZ PP CC 491 Acknowledgemen s: Ou hanks o Jean-Michel Bé enge (IHU Médi e anée In ec io) and D . 492 Phillip Kau man (Uni e si y o Flo ida) o acili a ing access o some pic u es o leas. 493 494 495 496 497 498 Figu es 499 Figu e 1: 1. Pulex i i ans emale, 2. Echidnophaga gallinacea emale (Koehle e al., 1991), 500 3. C enocephalides elis elis emale, 4. C enocephalides canis emale, 5. Xenopsylla cheopis 501 male, 6. A chaeopsylla e inacei emale. Pho og aphs o species (excep ing E. gallinacea 502 emale) we e aken using Nikon mic oscope equipped wi h a came a lucid sys em and a 503 pho omic oscope a he Uni e si y o Se ille, Se ille, Spain 504 505 Re e ences: 506 Ande son, B.E., Neuman, M.A., 1997. Ba onella spp. as eme ging human pa hogens. Clin. Mic obiol. 507 Re . 10, 203–219. 508 Angelakis, E., Medianniko , O., Pa ola, P., Raoul , D., 2016. Ricke sia elis: The Complex Jou ney o 509 an Eme gen Human Pa hogen. T ends Pa asi ol. 32, 554–564. 510 h ps://doi.o g/10.1016/j.p .2016.04.009 511 An onelli, P.L., Seymou , R.M., 1988. A model o myxoma osis based on ho monal con ol o abbi - 512 lea ep oduc ion. IMA J. Ma h. Appl. Med. Biol. 5, 65–80. 513 Audouin-Rouzeau, F., 2003. les chemins de la pes e. Le a , la puce e l’homme. P esses uni e si ai es 514 de Rennes. 515 Azzag, N., Haddad, N., Du and, B., Pe i , E., Ammouche, A., Chomel, B., Boulouis, H.-J., 2012. 516 Popula ion s uc u e o Ba onella henselae in Alge ian u ban s ay ca s. PloS One 7, e43621. 517 h ps://doi.o g/10.1371/jou nal.pone.0043621 518 Bahga , I.M., 2013. Mon hly abundance o oden and hei ec opa asi es in newly se led a eas, eas 519 o lakes, Ismailia Go e no a e, Egyp . J. Egyp . Soc. Pa asi ol. 43, 387–398. 520 Beaucou nu, J.-C., Launay, H., 1990. Les puces (Siphonap e a) de F ance e du bassin médi e anéen 521 occiden al, Faune de F ance. Féde a ion ancaise des socié és de sciences na u elles, 57, ue 522 Cu ie , 75231 Pa is Cedex 05. 523 Beaucou nu, J.C., Ménie , K., 1998. [The genus C enocephalides S iles and Collins, 1930 524 (Siphonap e a, Pulicidae)]. Pa asi e Pa is F . 5, 3–16. 525 h ps://doi.o g/10.1051/pa asi e/1998051003 526 Belkhi ia, J., Chomel, B.B., Ben Hamida, T., Kas en, R.W., S uckey, M.J., Fleischman, D.A., Ch is ophe , 527 M.M., Boulouis, H.-J., Fa e , T.B., 2017. P e alence and Po en ial Risk Fac o s o Ba onella 528 In ec ion in Tunisian S ay Dogs. Vec o Bo ne Zoono ic Dis. La chm . N 17, 388–397. 529 h ps://doi.o g/10.1089/ bz.2016.2039 530 Bel ho , J.R., Be nha d , S.A., Ball, C.L., G egg, M., Johnson, D.H., Ke e ling, R., P ice, E., Tinke , J.K., 531 2015. Bu owing Owls, Pulex i i ans, and Plague. Vec o Bo ne Zoono ic Dis. La chm . N 15, 532 556–564. h ps://doi.o g/10.1089/ bz.2015.1772 533 Be he a , E., Bekhoucha, S., Choug ani, S., Razik, F., Duchemin, J.B., Hou i, L., Deha ib, L., Fayolle, C., 534 Mak e oug ass, B., Dali-Yahia, R., Bellal, R., Belhab i, L., Chaieb, A., Tikhomi o , E., Ca niel, E., 535 2007. Plague eappea ance in Alge ia a e 50 yea s, 2003. Eme g. In ec . Dis. 13, 1459–1462. 536 h ps://doi.o g/10.3201/eid1310.070284 537 Bessas, A., Leulmi, H., Bi am, I., Zaidi, S., Ai -Oudhia, K., Raoul , D., Pa ola, P., 2016. Molecula 538 e idence o ec o -bo ne pa hogens in dogs and ca s and hei ec opa asi es in Algie s, 539 Alge ia. Comp. Immunol. Mic obiol. In ec . Dis. 45, 23–28. 540 h ps://doi.o g/10.1016/j.cimid.2016.01.002 541 Beu el, R.G., F ied ich, F., Whi ing, M.F., 2008. Head mo phology o Cau inus (Bo eidae, Mecop e a) 542 and i s phylogene ic implica ions. A h opod S uc . De . 37, 418–433. 543 h ps://doi.o g/10.1016/j.asd.2008.02.002 544 Bi am, I., Baziz, B., Rolain, J.-M., Belkaid, M., Raoul , D., 2006a. Zoono ic ocus o plague, Alge ia. 545 Eme g. In ec . Dis. 12, 1975–1977. h ps://doi.o g/10.3201/eid1212.060522 546 Bi am, I., Di ma , K., Pa ola, P., Whi ing, M.F., Raoul , D., 2010. Fleas and lea-bo ne diseases. In . J. 547 In ec . Dis. 14, e667–e676. h ps://doi.o g/10.1016/j.ijid.2009.11.011 548 Bi am, I., Pa ola, P., De La C uz, K.D., Ma sumo o, K., Baziz, B., Rolain, J.-M., Belkaid, M., Raoul , D., 549 2006b. Fi s molecula de ec ion o Ricke sia elis in leas om Alge ia. Am. J. T op. Med. 550 Hyg. 74, 532–535. 551 Boudebouch, N., Sa ih, M., Beaucou nu, J.-C., Ama ouch, H., Hassa , M., Raoul , D., Pa ola, P., 2011. 552 Ba onella cla idgeiae, B. henselae and Ricke sia elis in leas om Mo occo. Ann. T op. 553 Med. Pa asi ol. 105, 493–498. h ps://doi.o g/10.1179/1364859411Y.0000000038 554 Boudebouch, N., Sa ih, M., Chakib, A., Fadili, S., Boumzeb a, D., Zouiz a, Z., Mahadji, B.A., Ama ouch, 555 H., Raoul , D., Fou nie , P.-E., 2017. Blood Cul u e-Nega i e Endoca di is, Mo occo. Eme g. 556 In ec . Dis. 23, 1908–1909. h ps://doi.o g/10.3201/eid2311.161066 557 Bough on, R.K., A well, J.W., Schoech, S.J., 2006. An in oduced gene alis pa asi e, he s ick igh lea 558 (echidnophaga gallinacea), and i s pa hology in he h ea ened lo ida sc ub-jay (aphelocoma 559 coe ulescens). J. Pa asi ol. 92, 941–948. h ps://doi.o g/10.1645/GE-769R.1 560 Bouhsi a, E., Fe andez, Y., Liu, M., F anc, M., Boulouis, H.-J., Bi ille, F., 2013a. C enocephalides elis 561 an in i o po en ial ec o o i e Ba onella species. Comp. Immunol. Mic obiol. In ec . Dis. 562 36, 105–111. h ps://doi.o g/10.1016/j.cimid.2012.10.004 563 Bouhsi a, E., F anc, M., Boulouis, H.-J., Jacquie , P., Raymond-Le on, I., Liéna d, E., 2013b. 564 Assessmen o pe sis ence o Ba onella henselae in C enocephalides elis. Appl. En i on. 565 Mic obiol. 79, 7439–7444. h ps://doi.o g/10.1128/AEM.02598-13 566 B ossolle , J., 1990. [The disco e y o Ye sinia pes is]. Re . P a . 40, 1034–1036. 567 B own, L.D., Macaluso, K.R., 2016. Ricke sia elis, an Eme ging Flea-Bo ne Ricke siosis. Cu . T op. 568 Med. Rep. 3, 27–39. h ps://doi.o g/10.1007/s40475-016-0070-6 569 Bu le , T., 2014. Plague his o y: Ye sin’s disco e y o he causa i e bac e ium in 1894 enabled, in he 570 subsequen cen u y, scien i ic p og ess in unde s anding he disease and he de elopmen o 571 ea men s and accines. Clin. Mic obiol. In ec . 20, 202–209. h ps://doi.o g/10.1111/1469- 572 0691.12540 573 Cabanel, N., Lecle cq, A., Chenal-F ancisque, V., Annaja , B., Raje ison, M., Bekkhoucha, S., Be he a , 574 E., Ca niel, E., 2013. Plague ou b eak in Libya, 2009, un ela ed o plague in Alge ia. Eme g. 575 In ec . Dis. 19, 230–236. h ps://doi.o g/10.3201/eid1902.121031 576 Chomel, B.B., 1996. Ca -sc a ch disease and bacilla y angioma osis. Re . Sci. Tech. In . O . Epizoo . 577 15, 1061–1073. 578 Chomel, B.B., Kas en, R.W., 2010. Ba onelloses, an inc easingly ecognized zoonosis. J. Appl. 579 Mic obiol. 109, 743–750. h ps://doi.o g/10.1111/j.1365-2672.2010.04679.x 580 Coulaud, P.-J., Lepola d, C., Bechah, Y., Be enge , J.-M., Raoul , D., Ghigo, E., 2014. Hemocy es om 581 Pediculus humanus humanus a e hos s o human bac e ial pa hogens. F on . Cell. In ec . 582 Mic obiol. 4, 183. h ps://doi.o g/10.3389/ cimb.2014.00183 583 D'O enzio, E., Lemaî e, N., B oua , C., e al. 2018. Plague: B idging gaps owa ds be e disease 584 con ol. Med. Mal. In ec . 48, 307-317. doi:10.1016/j.medmal.2018.04.393 585 Dahesh, S.M.A., Mikhail, M.W., 2016. SURVEILLANCE OF TRYPANOSOMA SPP OF RODENTS AND 586 STUDIES IN THEIR TRANSMISSION PROBABILITY BY FLEAS IN SOME RURAL EGYPTIAN AREAS. 587 J. Egyp . Soc. Pa asi ol. 46, 157–166. 588 Da is, D.H., 1953. Plague in A ica om 1935 o 1949; a su ey o wild oden s in A ican e i o ies. 589 Bull. Wo ld Heal h O gan. 9, 665–700. 590 Dieme, C., Bechah, Y., Socolo schi, C., Audoly, G., Be enge , J.-M., Faye, O., Raoul , D., Pa ola, P., 591 2015. T ansmission po en ial o Ricke sia elis in ec ion by Anopheles gambiae mosqui oes. 592 P oc. Na l. Acad. Sci. U. S. A. 112, 8088–8093. h ps://doi.o g/10.1073/pnas.1413835112 593 Doble , G., P e e , M., 2011. Fleas as pa asi es o he amily Canidae. Pa asi . Vec o s 4, 139. 594 h ps://doi.o g/10.1186/1756-3305-4-139 595 D ancou , M., Maina di, J.L., B ouqui, P., Vandenesch, F., Ca a, A., Lehne , F., E ienne, J., Golds ein, 596 F., Aca , J., Raoul , D., 1995. Ba onella (Rochalimaea) quin ana endoca di is in h ee 597 homeless men. N. Engl. J. Med. 332, 419–423. 598 h ps://doi.o g/10.1056/NEJM199502163320702 599 D ancou , M., Raoul , D., 2018. Value o Ma hema ical Models o Epidemics: he Plague Pa adigm. 600 Clin. Mic obiol. In ec . O . Publ. Eu . Soc. Clin. Mic obiol. In ec . Dis. 601 h ps://doi.o g/10.1016/j.cmi.2018.08.014 602 D ancou , M., Raoul , D., 2016. Molecula his o y o plague. Clin. Mic obiol. In ec . O . Publ. Eu . 603 Soc. Clin. Mic obiol. In ec . Dis. 22, 911–915. h ps://doi.o g/10.1016/j.cmi.2016.08.031 604 D yden, M.W., Rus , M.K., 1994. The ca lea: biology, ecology and con ol. Ve . Pa asi ol. 52, 1–19. 605 Du den, L.A., T aub, R., 2002. Medical and Ve e ina y En omology. GARY MULLEN, LANCE DURDEN. 606 El Hamzaoui, B., La oche, M., Alme as, L., Bé enge , J.-M., Raoul , D., Pa ola, P., 2018. De ec ion o 607 Ba onella spp. in leas by MALDI-TOF MS. PLoS Negl. T op. Dis. 12, e0006189. 608 h ps://doi.o g/10.1371/jou nal.pn d.0006189 609 Els on, D.M., 2001. Wha ’s ea ing you? Echidnophaga gallinacea ( he s ick igh lea). Cu is 68, 250. 610 Faccini-Ma ínez, Á.A., Má quez, A.C., B a o-Es upiñan, D.M., Calix o, O.-J., López-Cas illo, C.A., 611 Bo e o-Ga cía, C.A., Hidalgo, M., Cue o, C., 2017. Ba onella quin ana and Typhus G oup 612 Ricke siae Exposu e among Homeless Pe sons, Bogo á, Colombia. Eme g. In ec . Dis. 23, 613 1876–1879. h ps://doi.o g/10.3201/eid2311.170341 614 F anc, M., 1994a. [Fleas and me hods o con ol]. Re . Sci. Tech. In . O . Epizoo . 13, 1019–1037. 615 F anc, M., 1994b. [Lice and me hods o con ol]. Re . Sci. Tech. In . O . Epizoo . 13, 1039–1051. 616 Gague e, E., P elaud, P., 2006. Guide p a ique de de ma ologie canine, Kalianxis. ed. 617 Gas ellie , L., Lan e nie , F., Ren oisé, A., Ri iè e, S., Raoul , D., Lo hola y, O., Lecui , M., 2015. 618 None up i e e e e ealing mu ine yphus in a a ele e u ning om Tunisia. J. T a el 619 Med. 22, 67–69. h ps://doi.o g/10.1111/j m.12154 620 Gillespie, J.J, Beie , M.S., Rahman, M.S., Amme man, N.C., Shallom, J.M., Pu kayas ha. A-, e al. 2007. 621 Plasmids and icke sial e olu ion: insigh om Ricke sia elis. PLoS ONE. 2:e266. doi: 622 10.1371/jou nal.pone.0000266 623 G ácio, A.J.D.S., G ácio, M.A.A. 2017. Plague: A Millena y In ec ious Disease Reeme ging in he XXI 624 Cen u y. Biomed. Res. In . 2017:5696542. doi:10.1155/2017/5696542 625 G a z, D.N., 1999. RODENT RESERVOIRS & FLEA VECTORS OF NATURAL FOCI OF PLAGUE 34. 626 Guine , F., Ca niel, E., 2008. [Should we s ill ea he plague oday?]. Med. Sci. MS 24, 865–868. 627 h ps://doi.o g/10.1051/medsci/20082410865 628 Ha ey, R.G., 1990. De ma i is in a ca associa ed wi h Spilopsyllus cuniculi. Ve . Rec. 126, 89–90. 629 Hinnebusch, B.J., Bland, D.M., Bosio, C.F., Ja e , C.O., 2017. Compa a i e Abili y o O opsylla 630 mon ana and Xenopsylla cheopis Fleas o T ansmi Ye sinia pes is by Two Di e en 631 Mechanisms. PLoS Negl. T op. Dis. 11, e0005276. 632 h ps://doi.o g/10.1371/jou nal.pn d.0005276 633 Ho nok, S., Föld á i, G., Rigó, K., Meli, M.L., Tó h, M., Molná , V., Gönczi, E., Fa kas, R., Ho mann- 634 Lehmann, R., 2014. Vec o -bo ne agen s de ec ed in leas o he no he n whi e-b eas ed 635 hedgehog. Vec o Bo ne Zoono ic Dis. La chm . N 14, 74–76. 636 h ps://doi.o g/10.1089/ bz.2013.1387 637 Houhamdi, L., Pa ola, P., Raoul , D., 2005. [Lice and lice-bo ne diseases in humans]. Med. T op. Re . 638 Co ps San e Colon. 65, 13–23. 639 Iannino, F., Salucci, S., Di P o ido, A., Paolini, A., Ruggie i, E., 2018. Ba onella in ec ions in humans 640 dogs and ca s. Ve . I al. 54, 63–72. h ps://doi.o g/10.12834/Ve I .398.1883.2 641 Jiang, J., Maina, A.N., Knobel, D.L., Clea eland, S., Laudisoi , A., Wambu u, K., e al. 2013. Molecula 642 de ec ion o Ricke sia elis and Candida us Ricke sia asemboensis in leas om human 643 habi a s, Asembo, Kenya. Vec o Bo ne Zoono ic Dis. 13, 550–558. doi: 644 10.1089/ bz.2012.1123 645 Kaal, J.F., Bake , K., To ge son, P.R., 2006. Epidemiology o lea in es a ion o uminan s in Libya. Ve . 646 Pa asi ol. 141, 313–318. h ps://doi.o g/10.1016/j. e pa .2006.05.034 647 Ka imi, Y., Fa hang-Azad, A., 1974. [Pulex i i ans, a human lea in he plaque in ec ion ocus a 648 Gene al Mobu u Lakd egion ( o me ly Lake Albe ): epidemiologic signi icance]. Bull. Wo ld 649 Heal h O gan. 50, 564–565. 650 Ke ni , T., Leulmi, H., Socolo schi, C., Be enge , J.-M., Lepidi, H., Bi am, I., Rolain, J.-M., Raoul , D., 651 Pa ola, P., 2014. Acquisi ion and exc e ion o Ba onella quin ana by he ca lea, 652 C enocephalides elis elis. Mol. Ecol. 23, 1204–1212. h ps://doi.o g/10.1111/mec.12663 653 Ke ni , T., S a o d, K., Coles, G.C., Bi am, I., Papa, K., Chia oni, J., Raoul , D., Pa ola, P., 2015. 654 Responses o a i icially ea ed ca leas C enocephalides elis elis (Bouché, 1835) o 655 di e en mammalian bloods. Med. Ve . En omol. 29, 171–177. 656 h ps://doi.o g/10.1111/m e.12100 657