UNCORRECTED PROOFZ 389 445 Continued

View metadata, citation and similar papers at core.ac.uk brought to you by CORE DTD 5 ARTICLE IN PRESS provided by Repositorio Institucional de la Universidad de Alicante

1 57 2 58 3 59 Micron xx (xxxx) 1–12 4 60 www.elsevier.com/locate/micron 5 61 6 62 7 63 8 Comparative morphology of early stages of two Mediterranean 64 9 65 10 Sarcophaga Meigen, 1826 (Diptera; Sarcophagidae) and 66 11 67 12 a review of the feeding habits of Palaearctic species 68 13 69 14 Salima Pe´rez-Moreno, M. Angeles Marcos-Garcıá*, Santos Rojo 70 15 71 16 Instituto de la Biodiversidad (CIBIO), Universidad de Alicante, E-03080 Alicante, Spain 72 17 Received 8 May 2005; revised 29 July 2005; accepted 30 July 2005 73 18 74 19 75 20 Abstract 76 21 77 22 The third instar larva of Sarcophaga hirticrus Pandelle´, 1896 and Sarcophaga javita (Peris, Gonza´lez-Mora and Mingo, 1998) are 78 23 described and figured for the first. The use of scanning electron microscopy (SEM) has been demonstrated as an effective tool for determining 79 differences at the specific level, and is here applied. The two species are distinguished from other Sarcophaga spp. and the principal 24 80 diagnostic character states are illustrated and discussed. Comparative information on immature stages morphology of the described 25 81 Palaearctic Sarcophaga species and its feeding habits are compiled and provided in a tabulated form. 26 82 q 2005 Elsevier Ltd. All rights reserved. 27 83 28 Keywords: Sarcophagidae; Immature stages; Larval morphology; Ultrastructure; Breeding habits 84 29 85 30 86 31 1. Introduction as being parasitic in vertebrates (mammals, turtles) 87 32 (Dahlem, 1991; Valle de Sales et al., 2004). 88 33 The immature stages of the majority of dipterous families In some cases, detailed descriptions of larvae implicated 89 34 remain poorly known (Hennig, 1968). In the case of the in traumatic human myiasis (Colwell and O’Connor, 2000; 90 35 Sarcophagidae relatively few papers have appeared dedi- Zumpt, 1965) or found infesting mummified human remains 91 36 cated to the larval morphology of the family (Aspoas, 1991; have been published (Sukontason et al., 2003), but in these 92 37 Ebejer, 2000; Kirk-Spriggs, 1999, 2000, 2003; Meńdez and cases the descriptions have not been attributed to any named 93 38 Pape, 2002; Zumpt, 1965). The larval stages of many species, as adults were not obtained. Therefore, the breeding 94 39 species of Sarcophagidae are necrophagous and for this habits should be related to morphological and taxonomic 95 40 reason those species termed ‘flesh-flies’ are significant in knowledge in order to improve biological and practical 96 41 forensic entomology, being second only to the Calliphoridae considerations. 97 42 (Diptera) in terms of their usefulness. Precise knowledge Sarcophagids larvae are easily recognised at family and 98 43 and precise diagnoses of their immature stages therefore generic level, but are morphologically remarkably similar 99 44 have a very practical application in estimating the post- subgenerically and inter-specifically (Aspoas, 1991). The 100 45 mortem interval (Sukontason et al., 2003; Wells et al., use of scanning electron microscopy (SEM) to observe and 101 46 2001). Other species of Sarcophaginae and Miltogramminae photograph the morphology of dipterous larvae has enabled 102 47 103 are predatory on invertebrates (e.g. other insects, snails, observation of a suite of intra-specific differences (Colwell, 48 104 earthworms, scorpions, crabs, etc.) (Meńdez and Pape, 1989), and to characterise some aspects (pseudocephalon, 49 105 2002; Pape, 1987), and several species have been reported spinules, anterior and posterior spiracles, spiracular setae, 50 106 rim of spiracular atrium tubercles and sensilia) not easily 51 107 resolved with light microscopy (Aspoas, 1991). For this 52 108 * CorrespondingUNCORRECTED authorTel.: C34 965 903400; fax: C34 965 903815. reason Cantrell PROOF (1981), indicates that it is important to 53 109 ´ 54 E-mail address: [email protected] (M.A. Marcos-Garcıa). describe the number of papillae forming the anterior 110 spiracles, the shape of the peritreme of the posterior 55 0968-4328/$ - see front matter q 2005 Elsevier Ltd. All rights reserved. 111 56 doi:10.1016/j.micron.2005.07.013 spiracles and the cephalopharingeal skeleton. 112

JMIC 869—6/9/2005—02:48—-[-no entity-]-—163450—XML MODEL 5 – pp. 1–12 DTD 5 ARTICLE IN PRESS

2 S. Pe´rez-Moreno et al. / Micron xx (xxxx) 1–12

113 Pape et al. (2002) cite 73 species of the genus approximately 24 h and were preserved in glycerine until 169 114 Sarcophaga as occurring in the Iberian Peninsula, but use. Measurements were made using a graticule mounted in 170 115 detailed morphological descriptions of larvae based on SEM an eye piece of a binocular microscope. 171 116 have been published for only five of these, namely: Stereoscan micrographs were taken with SEM HITACHI 172 117 Sarcophaga africa (Wiedemann, 1824) (as S. cruentata S3000N operated at 20 kV. Specimens of immature stages 173 118 Meigen, 1826), Sarcophaga crassipalpis Macquart, 1838, of S. hirticrus and S. javita are deposited in the 174 119 Sarcophaga dux Thomson, 1869, Sarcophaga exuberans Entomological Collection of the University of Alicante 175 120 Pandelle´, 1896 and Sarcophaga tibialis Macquart, 1851. (CIBIO). 176 121 Sarcophaga hirticrus Pandelle´, 1896 and Sarcophaga The character states examined are as follows: the 177 122 javita (Peris, Gonza´lez-Mora and Mingo, 1998) occur pseudocephalon, the cephalopharyngeal skeleton, the 178 123 sympatrically in southern Spain, where they are relatively spinules, the anterior spiracles, the spiracular atrium and 179 124 abundant. Both species have been occasionally collected the posterior spiracles. 180 125 together from the terrestrial snail Otala punctata (Mu¨ller, Taxonomic nomenclature in this paper follows Pape 181 126 1777) (Pe´rez-Moreno, 2004). (1996). Larval terminology used in this paper follows 182 127 In this study, the 3rd instar larvae of S. hirticrus and S. Teskey (1981) and Courtney et al. (2000) for ventral organ. 183 128 javita are described for the first time. Numerous features of 184 129 the 3rd instar larvae of the two species are examined by use 185 130 of SEM, in order to determine the presence of morphologi- 3. Results 186 131 cal character states of potential taxonomic value. 187 132 A table is provided which compiles published infor- 3.1. Sarcophaga hirticrus Pandelle´, 1896 188 133 mation about described larvae of Sarcophaga species and 189 134 feeding habits (Table 1). 3.1.1. Overall appearance (Fig. 1(A)) 190 135 Newly moulted larvae are creamy white in colour. 191 136 Length 9.85G0.86 mm, maximum width 3.63G0.31 mm 192 137 2. Materials and methods (nZ10). 193 138 Larvae elongated, sub-cylindrical in cross-section with a 194 139 All larval stages of S. hirticrus and S. javita were flattened ventral surface, truncated posteriorly and tapering 195 140 obtained from a laboratory colony maintained over several toward the anterior extreme. 196 141 generations under constant conditions of 25 8C temperature Posterior surface of anal segment with a distinct cavity, 197 142 and 70–85% relative humidity, and a photoperiod of 15:9 h which contains the posterior spiracles. The surface of 198 143 L:D. The colony was originated from field-collected thoracic and abdominal segments with bands of spinules of 199 144 material obtained in Mutxamel 30SYH203506 (Alicante, subtriangular form. The inter-band areas are devoid of 200 145 SE Spain) (Pe´rez-Moreno, 2004), during studies of the life spinules (Fig. 1(A)). 201 146 cycle of these species. Adult flies of S. hirticrus were placed Anal segment with two postanal tubercles with apical 202 147 into rearing cages and provided with a diet of sugar, water sensillae. Surface of the postanal tubercles covered with 203 148 and pig’s liver. The liver provided the protein meal for adult spinules in the half basal part. 204 149 female as well as a medium for larviposition. Larvae of S. 205 150 hirticrus were placed on a bed of a fine sand to facilitate the 3.1.2. Pseudocephalon (Fig. 1(B) and (C)) 206 151 pupation. Some of these puparia were left for 24/48 h in The pseudocephalon has a pair of small antennae (a) and 207 152 order to allow their sclerotisation before being killed by two palps (p) (Fig. 1(C)). Antennae appearing with two 208 153 freezing. In the case of S. javita, alive snails were provided segments, the apical (dome) in conical shape. The palps are 209 154 to the females in order to stimulate the larviposition. The mammeliform, with concentric ribbons at the apical 210 155 larvae and pupae were extracted with the help of a hand extreme where are placed five sesilla, the three posterior 211 156 needle of the snail’s shell. ones sited in the same protuberance and nominated 212 157 Third instar larvae were selected for preservation. These maxillary palpus sensilla (mxpp). 213 158 are distinguishable from larvae of other calyptrate families At both sides of the mouth, between the inferior part of 214 159 in that the majority of species possess three nearly vertical, the palps and the maxilla, appear extensive bands of 215 160 parallel, posterior spiracular openings, usually not orien- elongated, slightly corrugated overlapping oral ridges (or). 216 161 tated toward the opening in the peritreme. For permanent Ventral organs (vo) present. 217 162 preservation, larvae were killed by immersion in cold water The strong maxillae (m) are smooth, widely curved and 218 163 and slowly boiled for approximately four minutes to distend retractable (Fig. 1B). Buccal cavity displays a pair of 219 164 them. FollowingUNCORRECTED this they were preserved in 70% alcohol. sensilium (Fig. PROOF 1(C)). 220 165 Descriptions are based on preserved larvae and/or 221 166 puparia. Cephalonpharyngeal skeletons attached by a 3.1.3. Thorax (Fig. 1(D)) 222 167 membrane to the leading ventral edge of the interior of The prothorax is rounded by spinules one or two pointed 223 168 the puparia, were removed by immersion in water for and grouped in a variable number. Anterior spiracles (as) 224

JMIC 869—6/9/2005—02:48—-[-no entity-]-—163450—XML MODEL 5 – pp. 1–12 DTD 5 ARTICLE IN PRESS

S. Pe´rez-Moreno et al. / Micron xx (xxxx) 1–12 3

225 ) 281 226 282 227 283 228 284 229 285 230 286 Carrion, snail and insect Dead millipedes ter, internal parasite Myiasic,carrion Necrophagous Myiasic, necrophagous, parasitoids Dead animals and garbage casses Myiasic, carrion Myiasic, decomposing matter, feces, dead snails garbage parasite of moth 231 Necrophagous, carcasses, predators of lepidopterous pupae 287 232 288 continued on next page 233 ( 289 (F) Cas- 234 Pape, 290 (M&F) Horseﬂesh in laboratory (M&F) Decaying animal mat- (M&F) (M); (M&F) Chicken in laboratory (M&F) Flesh in laboratory, (M&F) Dead ﬁsh and mammals (M); (M&F) (M) (M); (M); (F) (F) 235 (F); 291 (F) (F); Blackith 236 Berner, 1973 292 Povolny and (F); a Bordas and

237 Zumpt, 1965 293 ˜ 238 294 tillo, 2001 et al., 1997 (F) Ishijima, 1967 Kirk-Spriggs, 1999 (M&F) Ishijima, 1967 Aspoas, 1991 Zumpt, 1965 Gonzalez-Mora, 2005 (M&F) Aspoas, 1991 Zumpt, 1965 1987 Ishijima, 1967 Cantrell, 1981 Sukontason et al., 2003 (M); Ishijima, 1967 Tumrasvin and Cano, 1979 (F) Ishijima, 1967 (F) Ishijima, 1967 Ishijima, 1967 Kano et al., 1951 (M) Dead animals, feces, Awad et al., 2003 (M&F); Verves, 1997 Ishijima, 1967 Pape, 1987 239 Ishijima, 1967 295 240 296 241 297 242 298 243 299 244 300 245 301 246 302 Material from References Host/feedind habits Kano et al., 1951 247 303 248 304 249 305 250 306 251 307 252 308 253 309 skeleton description (instar) 254 3rd instar 310 255 311

256 species 312 257 313 258 314

259 Sarcophaga 315 260 316 261 317 262 Ecdysial scar Cephalopharingeal 318 263 319 264 320 265 321 266 322 267 323 268 324 269 325 spiracles (3rd instar) 9–14/single row11–12 No Yes 3rd instar All instars Japan Australia 270 326 271 327 272 328 SEM 7–10/single row Indistinct 3rd instar Spain New data (M&F); LMLM 34–37 10/single row – No 3rd instar 3rd instar Japan Kalahari LM 40–44/irregular row No 3rd instar Japan SEM 8–10/single row – – South Africa SEM 11–13/single row – – South Africa LM 14–15/single row – 2nd and 3rd instar Spain Salon SEM 14–17/single row – All instars Thailand LM 9–14/single row – All instars Japan Kano et al., 1951 (M) Privies and animal car- LM 12–15/single row No 3rd instar Japan LM 32–36 No 3rd instar Japan LM 46–52/irregular rows No 3rd instar Japan LMLM 9–12/single row 32–38/irregular rows –SEM No 10–11/single row Yes All 3rd instars instar – Japan Egypt LM 48–54/irregular rows No 3rd instar Japan 273 329 274 330 Z

275 Kano, 331 Thom- Curran, 276 332 UNCORRECTEDMeigen, PROOF 277 333 , 1896 , 1896 ,1896 ´ ´ 278 ´ 334 (Pandelle,1896)) Robineau-Des- Zetterstedt, 1838 Macquart, 1838

279 Rovineauella sco- 335 ttcher,1912 tther, 1913 ¨ ¨ Z Sarcophaga horii 1953 Sarcophaga inzi 1934 Sarcophaga hirticrus Pandelle Table 1 Review of morphology of immature stages and feedingSpecies habits of Palaearctic T No. of lobes anterior Sarcophaga harpax Pandelle 1826) Sarcophaga aegyptica Salem, 1935 Sarcophaga africa (Wiedemann, 1824) ( S. cruentata son, 1869 Sarcophaga dux Sarcophaga brevicornis Ho, 1934 Sarcophaga caerules- cens ( Sarcophaga crassipalpis Sarcophaga exuberans Pandelle Sarcophaga alba (Schiner, 1868) Sarcophaga albiceps Meigen, 1826 Sarcophaga argyrostoma voidy, 1830 paria Sarcophaga caudagalli Bo 280 Sarcophaga antilope Bo 336

JMIC 869—6/9/2005—02:48—-[-no entity-]-—163450—XML MODEL 5 – pp. 1–12 DTD 5 ARTICLE IN PRESS

4 S. Pe´rez-Moreno et al. / Micron xx (xxxx) 1–12

337 393 338 394 339 395 340 396 341 397 342 398 ter, carcasses, garbage of lepidopterous oratory feaces, intestinal myiasis mals garbage, dead snails horseflesh in laboratory Human feces, carcasses, dead fish, snails predator oratory lished) in laboratory 343 mammals. Snails. Feaces. 399 344 400 345 401 346 402 (M&F) Dead animals, parasites (M&F) Animal matter in lab- (M&F) Human feces, dead ani- (M&F) (M&F) Dead animals (M&F) Fish, chicken and (M&F) Dead animals (M&F) (M&F) Animal matter in lab- (M&F) Horseflesh and chicken (M&F) Dung, carcasses (M&F); (M); (M&F) Decaying animal mat- (M&F); (F) 347 (F); 403 348 404 349 405 350 406 Ishijima, 1967 Ishijima, 1967 Ishijima, 1967 Ishijima, 1967 Cantrell, 1981 Seguy, 1941 Ishijima, 1967 Ishijima, 1967 Ishijima, 1967 Ishijima, 1967 Ishijima, 1967 Ishijima, 1967 Parashar and Rao, 1988 (F); Ishijima, 1967 Ishijima, 1967 351 Ishijima, 1967 Ishijima, 1967 Seguy, 1941 Blackith et al, 1997 (F) 407 352 408 353 409 354 410 355 411 356 412 357 413 358 414 Material from References Host/feedind habits 359 415 360 416 361 417 362 418 363 419 364 420 365 421 3rd instar 366 skeleton description (instar) 422 367 423 368 424 369 425 370 426 371 427 372 428 373 429 374 Ecdysial scar Cephalopharingeal 430 375 431 376 432 377 433 378 434 379 435 380 436 381 437 24–26/two rows28 No Yes 3rd instar All instars Japan Australia spiracles (3rd instar) 16–22/single row 3rd instar 382 438 383 439 384 440 LM 32–36 No 3rd instar Japan LM 46–49/irregular rows No 3rd instar Japan LM 24–30/two rows No All instars Japan Kano et al., 1951 (M) Carcasses, garbage, LM 28–34/two rows No 3rd instar Japan LM 24–28/two rows No All instars Japan Kano et al., 1951 (M) Myiasic, necrophagous, LMLM 14–16/single row 24–28 – No 3rd instar 3rd instar Japan Japan LM 42–46/irregular rows No 3rd instar Japan LM 28–30/two rows No 3rd instar Japan LM 16–17/single row No 3rd instar Japan LM 38–43/irregular rows No 3rd instar Japan SEM 12–14/single rowLM 34–38/two rows Indistinct No 3rd instar 3rd instar Spain Japan New data (M) Snail predator (unpub- LM 32–36/irregular rows No 3rd instar Japan LM 12–15/single row No All instars Japan Kano et al., 1951 (M) Myiasic; birds and 385 441 386 442 a ) 387 a 443

388 Tricho- 444 UNCORRECTED PROOFZ 389 445 continued

390 Hori, 1954 446 (Rohdendorf, Kano, 1950 391 Ho,1938 ( 447 tther, 1913 ¨ Sarcophaga schuetzei (Kramer,1909) Sarcophaga seniorwhi- tei Sarcophaga similis Meade, 1876 Sarcophaga peregrina Robineau-Desvoidy, 1830 Sarcophaga polystylata Ho, 1934 Sarcophaga pterygota Thomas, 1949 lioproctia ﬂavinervis (Senior-White,1924)) Sarcophaga septentrio- nalis Sarcophaga shirita- kaensis 1937) Table 1 ( Species T No. of lobes anterior Walker,1849 Bo Sarcophaga okazakii Kano, 1953 Sarcophaga orchidea Sarcophaga javita (Peris, Gonzalez-Mora & Mingo, 1998) Sarcophaga kagaensis Hori, 1954 392 Sarcophaga kawayuen- sis Sarcophaga melanura Meigen, 1826 Sarcophaga misera 448

JMIC 869—6/9/2005—02:48—-[-no entity-]-—163450—XML MODEL 5 – pp. 1–12 DTD 5 ARTICLE IN PRESS

S. Pe´rez-Moreno et al. / Micron xx (xxxx) 1–12 5

449 arising posterolaterally on the prothoracic segment and fan- 505 450 shaped (Fig. 1(D) pupa). Each anterior spiracle contains a 506 451 single row of papillae that number from 7 to 10. 507 452 The rest of the thoracic segments with an anterior band of 508 453 spinules encircling each segment. These bands are 509 described two taxa 454 constituted by seven crows of spinules without bifurcation. 510 Carrion 455 Myiasic, necrophagous, dead snails 511 . Column ‘T’, refers to the 456 3.1.4. Abdomen (Fig.1(A), (E)–(G)) 512 457 Abdominal segments with anterior spinule bands. These 513 Ishijima (1967)

458 Zumpt, 514 References

(M&F) (M&F) Chicken in laboratory bands encircle the segments. The inter-band areas are (M); 459 (F); devoid of spinules. 515 460 The distribution of the abdominal spinule bands is 516 s morphology and feeding habits are indicated 461 (F) different in dorsal and ventral position. In dorsal view the 517 462 518 Cantrell, 1981 Aspoas, 1991 Abasa, 1972 1965 Ishijima, 1967 Ishijima, 1967 spinules are posteriorly projected and form one simple band, 463 lacking spinules in the dorso-central area. These bands are 519 464 wider towards the last body segment. 520 465 In ventral view, each band is constituted by three 521 466 subbands of 3, 4–5, 3–4 rows of spinules alternately 522 467 disposed. The spinules of the ﬁrst subband are anteriorly 523 468 projected and the spinules of the second and third subbands 524 469 posteriorly. These spinules are one pointed (Fig. 1(E)). 525 470 In addition, there are rounded lateral areas in the spinules 526 471 of the sides of the segments. 527 472 Abdominal segments display small ﬂeshy protuberances 528 473 laterally (Fig. 1(A)). 529 474 Posterior spiracles on the last eleventh body segment are 530 475 set within a deep spiracular atrium (Fig. 1(G)). There are 12 531 476 532 Walker, 1849, we maintain this reference because it is probable that tubercles (t) located on the rim of the spiracular atrium 477 (Fig. 1(F)). Spinules distributed around the rim and 533 478 3rd instar 534 misera

. extending as far as the outer circumference of the ring 479 S formed by the tubercles. 535 480 Spiracles consisting of three elongate slits (s) oriented 536 481 vertically, with the openings disposed radially, each 537 482 surrounded by an incomplete sclerotized peritreme (p). 538 483 Ecdysial scar indistinct. A branch of spiracular hairs (sh), 539 484 540

and, when needed, between brackets, the name of the species used by the author cited in the column constituted by a single trunk with 2–3 branches, at both 485 as junior synonym of 541

– – South Africa sides of the slits. 486 The anal segment displays two postannal tubercles 542 487 whose surface is covered by spinules until the half. Each 543 488 Pape (1996) 544 Pape (1996) postanal tubercle has a sensilla in the apex. 489 545 490 546 491 3.1.5. Cephalopharyngeal skeleton 547 492 Mouthhook well developed (m); deeply pigmented, 548 493 sickle-shaped, pointed in the apex and basally square. 549 row 494 Dental sclerites (ds) present. Subhypostomal sclerite (ss) 550 495 middle moon-shaped from above. Intermediate sclerite 551 496 (hypopharyngeal sclerite, hs) short, wide with rounded 552 ttcher, 1913, is considered by LM 28 No All instars Australia SEM 15–21/irregular double LM 33–36/irregular rows No All instars Japan Kano et al., 1951 (M) Dead animals LM 14–17 No 3rd instar Japan ¨

497 Bo anterior and posterior projections, H-shaped from above and 553 498 not fused to the basal sclerite. Pharyngeal sclerite (ps) 554 . Z

499 it is indicated the valid name according to relatively bulky and heavily pigmented at middle, with a 555 orchidea .

500 misera parastomal bar (pb), slightly incurvated apically upwards. 556 S . S

UNCORRECTEDused for the description of the larvae; SEM (scanning electron microscopy) or LM (light microscope). In column ‘References’ the author that describe PROOF

501 Species Dorsal cornu (dc) wider and longer than ventral cornu. 557 Johnston and 502 Dorsal cornu with a narrow elongated window apically. 558

503 Although Ventral cornu (vc) approximately twice longer than wide, 559 a different to Sarcophaga taenionota (Wiedemann,1819) ( S. omega Tiegs, 1921) Sarcophaga tibialis Macquart, 1851 Sarcophaga tsushimae Senior-white, 1924 (‘M’ to morphology and ‘F’ to feeding habits). Sarcophaga ugamskii (Rohdendorf, 1937) In column 504 Technique apically subtruncated and with a small and dorsoapical oval 560

JMIC 869—6/9/2005—02:48—-[-no entity-]-—163450—XML MODEL 5 – pp. 1–12 DTD 5 ARTICLE IN PRESS

6 S. Pe´rez-Moreno et al. / Micron xx (xxxx) 1–12

561 617 562 618 563 619 564 620 565 621 566 622 567 623 568 624 569 625 570 626 571 627 572 628 573 629 574 630 575 631 576 632 577 633 578 634 579 635 580 636 581 637 582 638 583 639 584 640 585 641 586 642 587 643 588 644 589 645 590 646 591 647 592 648 593 649 594 650 595 651 596 652 597 Fig. 1. Sarcophaga hirticrus. (A) general ventral view, (B) mouthhook, (C) pseudocephalon (a; antenna, or; oral ridges, p; palp, vo; ventral organ), (D) anterior 653 spiracles (as, p; papillae) (puparium), (E) spinules, (F) spiracular atrium (puparium), (G) posterior spiracles (p; peritreme, s; slits, sh; spiracular hairs). 598 654 599 655 600 656 window. Dorsobasal lobe (dl) slightly marked near the apex 3.2.2. Pseudocephalon (Fig. 3(A), (B) and (D)) 601 657 The pseudocephalon has a pair of small antennae (a) in 602 of ventral cornu (Fig. 2). 658 603 dorsal position and two palps (p) in inferior position 659 (Fig. 3(A)). 604 3.2. Sarcophaga javita (Peris, Gonza´lez-Mora and 660 The antennae are located in a depression and are 605 Mingo, 1998) 661 606 formed by two segments, the apical (dome) sharpened at 662 607 its end. 663 608 3.2.1. Overall appearance The palps are mammeliform, with concentric ribbons at 664 G G 609 Length 11.46 1.73 mm, maximum width 4.6 the apical extreme. In the apical part there are three 665 Z 610 0.42 mm (n 10). Larvae are creamy white in colour, protuberance nominated maxillary palpus sensilla (mxpp) 666 611 elongated, sub-cylindrical in cross-section with a ﬂattened and exist two more situated at different levels (Fig. 3(D)). 667 612 ventral surface, truncated posteriorly and tapering toward Ventral organs (vo) present. 668 the anteriorUNCORRECTED extreme. At both sides PROOF of the mouth appear bands of little deep 613 669 614 The surface of thoracic and abdominal segments displays subparallel oral ridges (or). The base of the buccal cavity 670 615 bands of spinules rounded in form. The inter-band areas are displays a pair of sensilium each of them with one setae and 671 616 devoid of spinules. one hole (Fig. 3D). 672

JMIC 869—6/9/2005—02:48—-[-no entity-]-—163450—XML MODEL 5 – pp. 1–12 DTD 5 ARTICLE IN PRESS

S. Pe´rez-Moreno et al. / Micron xx (xxxx) 1–12 7

673 of these fleshy projections shows a sensilia. A group of 729 674 spinules appears at both sides of the insertion zone of the 730 675 fleshy projections. 731 676 732 677 3.2.5. Cephalopharyngeal skeleton 733 678 Cephalopharyngeal skeleton deeply pigmented. Mouth- 734 679 hook (m) sickle-shaped, pointed apically. Dental sclerite 735 680 (ds) present. Subhypostomal sclerite (ss) middle moon- 736 681 shaped from above. Intermediate sclerite (hypopharyngeal 737 682 sclerite, hs) short, longer than wide, H-shaped from above 738 683 and not fused to the basal sclerite, with anterior arms 739 684 tapering apically. Pharyngeal sclerite (ps) heavily pigmen- 740 685 ted with a pointed parastomal bar (pb). Extreme of 741 686 pharyngeal sclerite incurved downwards. Dorsal cornu 742 687 (dc) wider and longer than ventral cornu. Dorsal cornu 743 688 with a narrow elongated apical window and opened in the 744 689 apical extreme. Ventral cornu (vc) approximately twice 745 690 longer than wide and apically truncated. Window reduced to 746 691 a small split dorso-apically. Dorsobasal lobe (dl) slightly 747 692 Fig. 2. Cephalopharyngeal skeleton of S. hirticrus ((A) lateral view, (B) marked near the apex of ventral cornu (Fig. 5). 748 693 dorsal view); pb: parastomal bar, dc: dorsal cornu, vc: ventral cornu, ds: 749 694 dental sclerite, ps: pharyngeal sclerite, hs: hypopharyngeal sclerite, ss: 750 695 subhypostomal sclerite, dl: dorsobasal lobe, m: mouthhook. 4. Discussion 751 696 752 697 Within the Sarcophagidae, larvae of some species of the 753 698 3.2.3. Thorax (Fig. 3(C), (E)–(G)) genera Oxysarcodexia and Ravinia (Lopes and Leite, 1987), 754 699 The prothorax is anteriorly surrounded by a band of acute Sarcophaga (Aspoas, 1991; Kirk-Spriggs, 1999, 2000, 755 700 spinules (Fig. 3(C)), grouped in a variable number and some 2003; Sukontason et al., 2003) and Wohlfahrtia (Ruiz-Mar- 756 701 of them with two apical points. Anterior spiracles fan- tıńez et al., 1989), have been described using of SEM. The 757 702 shaped. Each anterior spiracle contains a single row of increase of this kind of study is a consequence that the 758 703 papillae that number from 12 to 14 (Fig. 3(G)). The rest of knowledge of the morphology of the immature stages 759 704 the thoracic segments, with an anterior band of spinules receiving more attention due to their use in medico-forensic 760 705 encircling each segment. These spinules have a single end entomology. 761 706 (Fig. 3(E) and (F)). The larvae of the two species described here, possess the 762 707 morphological character states defined by Dahlem (1991) 763 708 3.2.4. Abdomen (Figs. 3(H), 4(A)–(D)) for the Sarcophagidae: ‘deeply recessed posterior spiracles, 764 709 Abdominal segments with anterior spinule bands one mature larvae having three nearly vertical, parallel posterior 765 710 pointed and anteriorly projected. One of these bands spiracular slits arising from a ventral ecdysial scar (which is 766 711 narrower than the others and with the spinules posteriorly frequently indistinct or absent) and by the spiracular slits 767 712 projected. Inter-band areas are devoid of spinules. usually not pointing toward the opening in the peritreme’. 768 713 The posterior spiracles on the last eleventh body segment The cephalopharingeal skeleton is very similar in the two 769 714 are set within a deep spiracular atrium. There are 12 species studied as it is common in other saprophagous 770 715 tubercles (t) located on the rim of the atrium (Fig. 4(A)) diptera (Rotheray and Gilbert, 1999), but there are some 771 716 each one with an apical setae (Fig. 4(D)). Spiracular atrium differences. The mouthhooks of S. hirticrus are proportion- 772 717 tapestried internally with spinules finished in a hook, ally larger and more curved than those in S. javita. Other 773 718 sometimes grouped and none of them bifid (Fig. 4(C)). authors have noted a limit in the mouthhooks retraction into 774 719 These spinules extended only as far as the inner the cephalic segment in other species such as Wohlfahrtia 775 720 circumference of the ring of tubercles. Posterior spiracles magnifica (Ruiz-Martıńez et al., 1990). This character state 776 721 posterodorsal in the atrium, on a spiracular plate. Spiracles should be analysed in other Sarcophaga species. 777 722 consisting of three elongate slits (s), oriented vertically, with In S. hirticrus the ventral cornu of the basal sclerite has a 778 723 the openings disposed radially, each with an incomplete small window, which is reduced to a small split in S. javita, 779 724 sclerotized peritremeUNCORRECTED (p) (Fig. 3(H)). Ecdysial scar whereas the parastomal PROOF bar is straighter in S. javita. 780 725 indistinct. The presence of a central hook, as in W. magnifica,is 781 726 The anal segment (Fig. 4(B)) displays two postannals possibly consistent with their parasitic nature (Ruiz-Mar- 782 727 tubercles. The internal margin of these fleshy projections is tıńez, 1990). It should also be interesting to describe the 783 728 covered with little sharp-pointed spinules. The apical stream surface, ornamentation, shape and size of the mouthhooks in 784

JMIC 869—6/9/2005—02:48—-[-no entity-]-—163450—XML MODEL 5 – pp. 1–12 DTD 5 ARTICLE IN PRESS

8 S. Pe´rez-Moreno et al. / Micron xx (xxxx) 1–12

785 841 786 842 787 843 788 844 789 845 790 846 791 847 792 848 793 849 794 850 795 851 796 852 797 853 798 854 799 855 800 856 801 857 802 858 803 859 804 860 805 861 806 862 807 863 808 864 809 865 810 866 811 867 812 868 813 869 814 870 815 871 816 872 817 873 818 874 819 875 820 876 821 877 822 878 823 879 824 880 825 881 826 882 827 883 828 884 829 885 830 886 831 887 832 888 833 889 834 890 835 891 836 UNCORRECTED PROOF 892 837 893 838 894 Fig. 3. S. javita. Pseudocephalon: (A) general view (a; antenna, or; oral ridges, p; palp, vo; ventral organ), (B) oral cavity ampliﬁed, (C) prothoracic spinules, 839 895 (D) palp (mxpp; maxillary palpus sensilla). Thorax: (E) dorsal spinules, (F) spinules ampliﬁed, (G) anterior spiracle, (H) posterior spiracle (p; peritreme). 840 896

JMIC 869—6/9/2005—02:48—-[-no entity-]-—163450—XML MODEL 5 – pp. 1–12 DTD 5 ARTICLE IN PRESS

S. Pe´rez-Moreno et al. / Micron xx (xxxx) 1–12 9

897 953 898 954 899 955 900 956 901 957 902 958 903 959 904 960 905 961 906 962 907 963 908 964 909 965 910 966 911 967 912 968 913 969 914 970 915 971 916 972 917 973 918 974 919 975 920 976 921 977 Fig. 4. S. javita. (A) tubercles of external border of the spiracular atrium, (B) postanal tubercles (C) internal atrium spinules, (D) tubercle amplified showing a 922 978 setae. 923 979 924 detail in order to establish their possible relationship with and Tiegs, 1921), S. dux, Sarcophaga nodosa Engel, 1925, 980 925 feeding habits. In this sense Wohlfahrtia magnifica (Schiner, Sarcophaga inzi Curran, 1934, and in species of other 981 926 1862), S. dux, Sarcophaga argyrostoma Robineau-Des- genera such as Wohlfahrtia virgil (Walker, 1849) and 982 927 voidy, 1830 and S. hirticrus have an inferior cutting margin Oxysarcodexia confusa Lopes, 1946 (Leite and Lopes, 983 928 of the mouthhook (Ruiz-Martıńez, 1990; Sukontason et al., 1987; Walker, 1920). This indicates that the exclusive use of 984 929 2003; Awad et al., 2003) in order to assist its penetration this criterion is insufficient for a correct identification of the 985 930 into carrion or wounds of animals, whereas Ravinia belforti larvae, even at generic level. Kano and Sato (1951), indicate 986 931 show a mouthhook with delicate pits and ridges (Leite and in Sarcophaga a slight variability in the number of papillae 987 932 988 Souza, 1987). between individuals or even on each side of the same pupae. 933 989 In the pseudocephalon of S. hirticrus, the palps and the This variability also occurs in other genera such as 934 990 antennae, are situated on a conical prominence. This 935 991 morphology is also appreciated in other species such as 936 992 Sarcophaga forceps Blackith and Blackith, 1988 (Kirk-- 937 993 Spriggs, 2000), however, this prominence is less pro- 938 994 nounced in S. javita. Bearing in mind the differences found 939 995 940 in the pseudocefalon of the species of Sarcophagidae, it 996 941 would be interesting to illustrate this segment from the same 997 942 angle always in order to facilitate comparison between 998 943 species. 999 944 Although the morphology of the pseudocephalon has not 1000 945 usually been considered, according to Kirk-Spriggs (2003), 1001 946 the disposition and morphology of the oral ridges of the two 1002 947 species here described seem to be useful inter-specific 1003 948 character states. 1004 The numberUNCORRECTED of papillae in the anterior spiracles of these PROOF 949 1005 two species at L3 instar (7–14) is similar to that of other Fig. 5. S. javita cephalopharyngeal skeleton ((A) lateral view, (B) dorsal 950 view); pb: parastomal bar, dc: dorsal cornu, vc: ventral cornu, ds: dental 1006 951 species of the same genera such as Sarcophaga melanura, S. sclerite, ps: pharyngeal sclerite, hs: hypopharyngeal sclerite, ss: sub- 1007 952 crassipalpis, Sarcophaga africa, Sarcophaga iota (Johnston hypostomal sclerite, m: mouthhook. 1008

JMIC 869—6/9/2005—02:48—-[-no entity-]-—163450—XML MODEL 5 – pp. 1–12 DTD 5 ARTICLE IN PRESS

10 S. Pe´rez-Moreno et al. / Micron xx (xxxx) 1–12

1009 Wohlfahrtia, Goniophyto and Ravinia varying from 1 to 8 The immature stages of other species of Sarcopaga such 1065 1010 (Walker, 1937; Maurice et al., 1948; Kano and Sato, 1951; as Sarcophaga peregrina, S. melanura, S. africa, Sar- 1066 1011 Ishijima, 1967; Ruiz-Martıńez et al., 1990). However, the cophaga misera and S. tibialis have been described and 1067 1012 arrangement of the papillae in regular or irregular rows is related to snails and other feeding sources (Seguy, 1941; 1068 1013 constant at species level (Table 1) being numerous and Ishijima, 1967; Berner, 1973; Parashar and Rao, 1988; 1069 1014 disposed in several files in the Japanese species (Kano and Pe´rez-Moreno, 2004). Apparently, none of the character 1070 1015 Sato, 1951). states of Sarcophaga larvae and puparia seem to be 1071 1016 It is not possible to establish a clear relationship between correlated with particular features of the breeding habits. 1072 1017 the number and distribution of the papillae in the anterior In order to find a relationship between the feeding site and 1073 1018 spiracles and the feeding habits (Table 1). the larval morphology, similarities between those species 1074 1019 The arrangement of the spinules in the spiracular atrium should be sought in species with a specific feeding habit. 1075 1020 of S. hirticrus (as far as the outer circumference of the ring Unfortunately, feeding habits of the genus Sarcophaga are 1076 1021 of tubercles) is similar to that found in S. exuberans very varied and there are not enough detailed descriptions 1077 1022 (Aspoas, 1991), whereas in S. javita, S. tibialis and S. dux realised by SEM to reach a conclusion (Table 1). 1078 1023 (Aspoas, 1991; Sukontason et al., 2003) these spinules are Kirk-Spriggs (2003) considers that in the case of the 1079 1024 extended only as far as the inner circumference of the ring. necrophagous Sarcophaga namibia Reed, 1974 the exten- 1080 1025 The disposition and shape of the slits on the posterior sive overlapping oral ridges of the facial mask are consistent 1081 1026 spiracles as well as the presence of an ecdysial scar are a with saprophagy. The same character state occurs in S. 1082 1027 useful tool in the identification of the Sarcophaginae species hirticrus, also a necrophagous species. In this sense, it 1083 1028 (Cantrell, 1981), with these characters being repeatedly would be interesting to include the description of the oral 1084 1029 described. ridges in future descriptions. 1085 1030 Due to the uniformity of morphological structures, many The early stage characters in other Diptera Cyclorrhapha 1086 1031 more species would need to be examined before any generic as Syrphidae, have been informative phylogenetically 1087 because their larvae are interpreted to be conservative in 1032 separation could be undertaken on the basis of larval 1088 evolution (Rotheray and Gilbert, 1999; Pe´rez-Banõń et al., 1033 morphology (Cantrell, 1981). However, differences in some 1089 2003). The detailed description of early stage morphology 1034 characters of 3rd instar larva have been useful in the 1090 in Sarcophaga species using SEM, could also be used in the 1035 elaboration of keys at specific levels. The main characters 1091 future together with other character sets such as DNA 1036 used in the existing keys are: number of papillae in the 1092 1037 sequences to estimate Sarcophaga phylogeny. 1093 anterior spiracles; pigmentation, arrangement and hardness 1038 1094 of the spinules on the body segments and cephalopharyngeal 1039 1095 skeleton (James and Gassner, 1947). Kano and Sato (1951) 1040 1096 add the following characters: arrangement of papillae in the 5. Uncited references 1041 1097 anterior spiracles, tubercles on upper border of anal 1042 1098 segment, inner projections of peritreme and posterior Ali-khan, 1974. Barfoot, 1969. Beaver, 1972. Blackith et 1043 al., 1928. Coupland and Baker, 1994. Salonã Bordas and 1099 spiracles (width and height, scar). Ishijima (1967), 1044 Gońzalez-Mora, 2005. Tumrasvin and Kano, 1979. 1100 completes the keys adding: distance between posterior 1045 1101 spiracles and morphology of slits. After using SEM, new 1046 1102 characters have been incorporated into the species diag- 1047 Acknowledgements 1103 1048 nosis: morphology of tubercles on the atrium rim and size of 1104 the fleshy projections on the anal segment (Lopes and Leite, 1049 The authors are deeply grateful to T. Pape for the 1105 1050 1987); number of rows, orientation and density of spinules 1106 in the body segments and spiracular hairs in posterior information provided and his very useful comments. Thanks 1051 also to the anonymous referees for their useful suggestions. 1107 1052 spiracles (Aspoas, 1991). Awad et al. (2003) take into 1108 account new ultrastructure characters: sensillar numbers and We thank C. Pe´rez-Banõń for her help with electron 1053 microscopy techniques and A. Martıńez-Sańchez for her 1109 1054 types, sizes and locations of the antennal-maxillary sensory 1110 complex. advices about maintenance of colonies in the laboratory. 1055 Financial support was partially provided by the Spanish 1111 As a general reflection from the preceding compilation 1056 Ministerio de Medio Ambiente (No. 040/2002). 1112 1057 and the new data provided in our descriptions, it can be 1113 1058 concluded that the most useful morphological characters for 1114 1059 diagnosis at specific level are: the structures of both, anterior 1115 References 1060 and posterior spiracles, the morphology of the pseudoce- 1116 phalon (includingUNCORRECTED oral ridges, antenna, maxillary palp, PROOF 1061 1117 sensilla and ventral organ) as proposed by Kirk-Spriggs Abasa, R.O., 1972. Reproductive biology of Sarcophaga tibialis (Diptera: 1062 Sarcophagidae). II. Morphology of external and internal reproductive 1118 1063 (2003) and the morphology and distribution of the spinules organs, ovary growth, and oogenesis. Annals of the Entomological 1119 1064 and sesilla in the body segments. Society of America 65 (2), 400–405. 1120

JMIC 869—6/9/2005—02:48—-[-no entity-]-—163450—XML MODEL 5 – pp. 1–12 DTD 5 ARTICLE IN PRESS

S. Pe´rez-Moreno et al. / Micron xx (xxxx) 1–12 11

1121 Ali-khan, F.E.A., 1974. Two cases of human Sarcophaga (Diptera: flesh of a decomposing cowrie shell in Sulawesi, Indonesia. Studia 1177 1122 Sarcophagidae) myiasis in Quebec, with descriptions of the larvae. Dipterologica 7 (1), 125–131. 1178 Canadian Journal of Zoology 52, 643–647. Kirk-Spriggs, A.H., 2003. The immatures stages of Sarcophaga(Liosarco- 1123 1179 Aspoas, B.R., 1991. Comparative micromorphology of third instar larvae phaga) namibia Reed (Diptera: Sarcophagidae) from the southwestern 1124 and the breeding biology of some Afrotropical Sarcophaga (Diptera: seaboard of Africa. Cimbebasia 18, 39–47. 1180 1125 Sarcophagidae). Medical and Veterinary Entomology 5, 437–445. Leite, A.C.R., Lopes, H.S., 1987. Second contribution to the knowledge of 1181 1126 Awad, A., Abdel-Salam, S., El-Ela, R., Abdel-Aal, A., Mohamed, D., 2003. the larvae of the Raviniini (Diptera, Sarcophagidae) based on 1182 1127 Ultrastructure comparison of the sensory morphology of the first- and observations using scanning electron microscope. Memorias Do 1183 third-instar larvae of Parasarcophaga argyrostoma (Robineau-Des- Instituto Oswaldo Cruz 82, 219–226. 1128 1184 voidy) (Diptera: Sarcophagidae). Egyptian Journal of Biology [Online] Lopes, H.S., Leite, A.C.R., 1987. Third contribution to the knowledge of 1129 5, 148–154. the Raviniini (Diptera, Sarcophagidae), based on observations of the 1185 1130 Barfoot, S.D., 1969. Sarcophaga nigriventris and S. hirticrus (Diptera: larvae, using scanning electron microscope. Memorias Do Instituto 1186 1131 Calliphoridae) both bred from Helix aspersa Mu¨ller (Mollusca Oswaldo Cruz 82, 407–413. 1187 1132 Helicidae). Entomologist’s Monthly Megazine 105, 49. Maurice, T.J., Gassner, F.X., 1948. The immature stages of the fox 1188 1133 Beaver, R.A., 1972. Ecological studies on Diptera breeding indead snails. 1. maggot Wohlfahrtia opaca (Cog.). The Journal of Parasitology 33, 1189 Biology of the species found in Cepaea nemoralis (L.). The 241–244. 1134 1190 Entomologist 105, 41–52. Mendez, J., Pape, T., 2002. Biology and immature stages of Peckia gulo 1135 Berner, L., 1973. Sur le parasitisme des he´licide´s par les mouches du genre (Fabricius, 1805) (Diptera: Sarcophagidae). Studia Dipterologica 9, 1191 1136 Sarcophaga. Bulletin du Museúm d’histoire Naturelle de Marseille 33, 371–374. 1192 1137 87–94. Pape, T., 1987. The Sarcophagidae (Diptera) of Fennoscandia and 1193 1138 Blackith, R.M., Blackith, R.E., Pape, T., 1928. Taxonomy and Denmark. Fauna Entomologica Scandinavica 19, 1–203 (2 pl. Leiden 1194 systematics of the taxon Helicophagella Enderlein, 1928 (Diptera: and Copenhagen). 1139 1195 Sarcophagidae), with the description of a new species. Studia Pape, T., 1996. Catalogue of the Sarcophagidae of the world (Insecta: 1140 1196 Dipterologica 4, 383–434. Diptera). Memoirs of Entomology International 8, 1–558. 1141 Cantrell, B.K., 1981. The immature stages of some Australian Sarcopha- Pape, T., Gońzalez-Mora, D., Peris, S., 2002. Sarcophagidae. pp. 218–221. 1197 1142 ginae (Diptera: Sarcophagidae). Journal of the Australian Entomologi- In: Cata´logo de los Diptera de Espanã, Portugal y Andorra (Insecta). 1198 1143 cal Society 20, 237–248. Monografıás S.E.A. Miguel Carles-Tolra´ Hjorth-Andersen (Coord.), 1199 1144 Castillo, M., 2001. Artro´podos presentes en carronã de cerdos en la 323 pp. 1200 comarca de la Litera (Huesca). Boletin de la Sociedad Estomatologica Parashar, B.D., Rao, K.M., 1988. Biological studies of the flesh fly 1145 1201 Argentina 28, 133–140. Sarcophaga (Parasarcophaga) misera and its effects as a predator of the 1146 Colwell, D.D., 1989. Scanning electron microscopy of the posterior snail Indoplanorbis exustus. Entomophaga 33, 431–434. 1202 1147 spiracles of cattle grubs Hypoderma bovis and Hypoderma lineatum. Pe´rez-Banõń, C., Rojo, S., Stahls, G., Marcos-Garcıá, M.A., 2003. 1203 1148 Medical and Veterinary Entomology 3 (4), 391–398. Taxonomy of European Eristalinus (Diptera: Syrphidae) based on 1204 1149 Colwell, D.D., O’Connor, M., 2000. Scanning electron microscopy of larval morphology and molecular data. European Journal of Entomol- 1205 1150 sarcophagid (Diptera) larvae recovered from a case of human cutaneous ogy 100, 417–428. 1206 myiasis. Journal of Medical Entomology 37, 854–859. Pe´rez-Moreno, S., 2004. Dı´pteros sarcofa´gidos (Diptera, Sarcophagidae) 1151 1207 Coupland, J.B., Baker, G., 1994. Host distribution, larviposition behaviour relacionados con gastero´podos terrestres (Mollusca, Gastropoda) en 1152 and generation time of Sarcophaga penicillata (Diptera: Sarcophagi- ecosistemas mediterrańeos. Boletıń de la Asociacioń Espanõla de 1208 1153 dae), a parasitoid of conical snails. In: Bulletin of entomological Entomologıá 28, 187–205. 1209 1154 Research, vol. 84. pp. 185–189. Povolny, D., Verves, Y., 1997. The Flesh-Flies of Central Europe (Insecta, 1210 1155 Courtney, G.W., Sinclair, B.J., Meier, R., 2000. 1.4. Morphology and Diptera, Sarcophagidae). SpixianaSuppl. 24, 1–260. 1211 terminology of Diptera larvae. In: Papp, L., Darvas, B. (Eds.), Rotheray, G.E., Gilbert, F.S., 1999. The phylogeny of palaearctic 1156 1212 Contributions to a Manual of Palaearctic Diptera, vol. 1. Science Syrphidae: evidence from larval stages. Zoological Journal of the 1157 Herald, Budapest, pp. 85–161. [vi]C7–978 pp. Linnaean Society 127, 1–112. 1213 1158 Dahlem, G.A., 1991. Sarcophagidae (Oestroidea). In: Stehr, F.W. (Ed.), Ruiz-Martıńez, I., Soler-Cruz, M.D., Benı´tez-Rodrı´guez, R., Dıáz-Lo´pez, 1214 1159 Immature Insects, vol. 2. Kendall/Hunt Publishing Company, Dubuque, M., Pe´rez-Jimeńez, J.M., 1989. Preparation of dipteran larvae for 1215 1160 IA, pp. 871–873. 975 pp. scanning electron microscopy with special reference to myasigen 1216 1161 Ebejer, M.J., 2000. Description of third instar larva and puparium of dipteran species. Scanning Microscopy 3, 387–390. 1217 Blaesoxipha calliste Pape (Diptera: Sarcophagidae). Studia Dipter- Ruiz-Martıńez, I., Soler-Cruz, M.D., Benı´tez-Rodrı´guez, R., Pe´rez- 1162 1218 ologica 7 (1), 121–124. Jimeńez, J.M., Adalid-Fuentes, C., Dıáz-Lo´pez, M., 1990. Scanning 1163 Hennig, W., 1968. Die Larvenformen der Dipteren, Einebersicht Ber Die Electron microscope study of W. magnı´fica (Schiner,1862) (Diptera: 1219 1164 Bisher Bekannten Jugendstadien Der Zweiflgeligen Inseckten. 3. Teil. Sarcophagidae). I. Structures with parasitic and possible taxonomic 1220 1165 Akademie Verlag, Berlin. [vii]C628 pp. meaning. Scanning Microcopy 4, 103–109. 1221 1166 Ishijima, H., 1967. Revision of the third stage larvae of synanthropic flies of Salonã Bordas, M.I., Gońzalez-Mora, D, 2005. Primera cita de 1222 Japan (Diptera: Anthomyiidae, Muscidae, Calliphoridae and Sarcopha- Liosarcophaga aegyptica (Salem, 1935) (Diptera, Sarcophagidae) de 1167 1223 gidae). Japanese Journal of Sanitary Zoology 18, 47–100. la Penıńsula Ibe´rica, con descripcioń de sus fases larvarias II y III, 1168 1224 James, M.T., Gassner, F.X., 1948. The immature stages of the fox maggot, pupario y adultos. Boletıń Sociedad Entomolo´gica Aragonesa 36, 1169 Wohlfahrtia opaca (Coq.). The Journal of Parasitology 33, 241–244. 251–255. 1225 1170 Kano, R., Tange, H., 1951. Notes on the flies of medical importance in Seguy, E., 1941. Etu´des sur les mouches parasites. 11 Calliphorides. 1226 1171 Japan (part II). The larvae of Sarcophaga known in Japan. Japanese Calliphorides (suite), Sarcophagines et Rhinophorides de l’Europe 1227 1172 Journal of Experimental Medicine 21, 115–131. occidentale et meridionales. Encycl. Ent. 21, 436. 1228 Kirk-Spriggs, A.H.,UNCORRECTED 1999. Female, immatures, and hymenopteran parasites Sukontason, K., Sukontason,PROOF K.L., Piangjai, S., 2003. Scanning 1173 1229 of Sarcophaga inzi Curran (Diptera: Sarcophagidae). Cimbebasia 15, electron microscopy of third-instar sarcophagid (Diptera: Sarco- 1174 65–70. phagidae) recovered from a mummified human corpse in Thailand. 1230 1175 Kirk-Spriggs, A.H., 2000. The immature stages of Sarcophaga forceps Revista do Instituto de Medicina Tropical de Sao Paulo 45, 1231 1176 Blackith and Blackith, 1988 (Diptera: Sarcophagidae), reared from the 95–98. 1232

JMIC 869—6/9/2005—02:48—-[-no entity-]-—163450—XML MODEL 5 – pp. 1–12 DTD 5 ARTICLE IN PRESS

12 S. Pe´rez-Moreno et al. / Micron xx (xxxx) 1–12

1233 Teskey, H.J., 1981. Morphology and terminology—larvae. In: McAlpine, (Diptera) parasitando Cebus apella (L., 1758) (Primate: Cebidae) no 1289 J.F., Peterson, B.V., Shewell, G.E., Teskey, H.J., Vockeroth, J.R., zoolo´gico de sorocaba, estado de Saõ paulo, Brasil. Entomol. Vect. 11 1234 1290 Wood, D.M. (Eds.), Manual of Nearctic Diptera Monograph of the (2), 317–321. 1235 Biosystematics Research Institute, No. 27, vol. 1, pp. 65–88. Walker, E.M., 1937. The larval stages of Wohlfahrtia vigil (Walker). The 1291 1236 Tumrasvin, W., Kano, R., 1979. Studies on medically important flies in Journal of the Parasitology 23, 163–174. 1292 1237 Thailand. VI. Reports on 48 species of sarcophagid flies, including the Wells, J.D., Pape, T., Sperling, F.A.H., 2001. DNA-based identification and 1293 1238 taxonomic keys (Diptera: Sarcophagidae). Bulletin of Tokyo Medical molecular systematics of forensically important Sarcophagidae 1294 1239 and Dental University 26, 149–179. (Diptera). Journal of Forensic Sciences 46, 1098–1102. 1295 Valle de Sales, K., Hidalgo-Friciello, R., Marinete-Amorim, T., Salles- Zumpt, F., 1965. Myasis in Man and Animals in the Old World. 1240 Gazeˆta, G., Serra-Freire, N.M., 2004. Relato de larva de sarcophagidae Butterworths, London. 1296 1241 1297 1242 1298 1243 1299 1244 1300 1245 1301 1246 1302 1247 1303 1248 1304 1249 1305 1250 1306 1251 1307 1252 1308 1253 1309 1254 1310 1255 1311 1256 1312 1257 1313 1258 1314 1259 1315 1260 1316 1261 1317 1262 1318 1263 1319 1264 1320 1265 1321 1266 1322 1267 1323 1268 1324 1269 1325 1270 1326 1271 1327 1272 1328 1273 1329 1274 1330 1275 1331 1276 1332 1277 1333 1278 1334 1279 1335 1280 1336 1281 1337 1282 1338 1283 1339 1284 UNCORRECTED PROOF 1340 1285 1341 1286 1342 1287 1343 1288 1344

JMIC 869—6/9/2005—02:48—-[-no entity-]-—163450—XML MODEL 5 – pp. 1–12