PUBLISHED BY THE AMERICAN MUSEUM OF NATURAL HISTORY CENTRAL PARK WEST AT 79TH STREET, NEW YORK, NY 10024 Number 3449, 67 pp., 26 ®gures, 6 tables August 23, 2004

Austroconops Wirth and Lee, a Lower of Biting Yet Living in Western Australia: a New , First Description of the Immatures and Discussion of Their Biology and Phylogeny (Diptera: )

ART BORKENT1 AND DOUGLAS A. CRAIG2

ABSTRACT

The eggs and all four larval instars of Austroconops mcmillani Wirth and Lee and A. annettae Borkent, new species, are described. The pupa of A. mcmillani is also described. Life cycles and details of behavior of each life stage are reported, including feeding by the aquatic larvae on microscopic organisms in very wet soil/detritus, larval locomotion, female adult biting habits on humans and kangaroos, and male adult swarming. Austroconops an- nettae Borkent, new species, is attributed to the ®rst author. Cladistic analysis shows that the two extant Austroconops Wirth and Lee species are sister species. Increasingly older fossil species of Austroconops represent increasingly earlier line- ages. Among extant lineages, Austroconops is the sister group of Leptoconops Skuse, and together they form the sister group of all other Ceratopogonidae. Kieffer is the sister group of Meigen ϩ Kieffer, and together they form the sister group of the Ceratopogoninae. Forcipomyia has no synapomorphies and may be paraphyletic in relation to Atrichopogon. Austroconops is morphologically conservative (possesses many plesiomorphic features) in each life stage and this allows for interpretation of a number of features within Ceratopogonidae and other . A new interpretation of Cretaceous fossil lineages shows that Austroconops, Leptoconops, Minyohelea Borkent, Jordanoconops

1 Royal British Columbia Museum, American Museum of Natural History, and Instituto Nacional de Biodiversidad. Address: 691-8th Ave. SE, Salmon Arm, BC, VIE 2C2, Canada ([email protected]). 2 Department of Biological Sciences, CW-405 Biological Sciences Centre, University of Alberta, Edmonton, AB, T6G 2E9, Canada ([email protected]).

Copyright ᭧ American Museum of Natural History 2004 ISSN 0003-0082 2 AMERICAN MUSEUM NOVITATES NO. 3449

Szadziewksi, Archiaustroconops Szadziewksi, and Fossileptoconops Szadziewksi form a monophyletic group. Within this assemblage Leptoconops and Minyohelea are sister groups and Austroconops and Jordanoconops are monophyletic (Austroconops is possibly paraphyletic in relation to Jordanoconops). All are considered to be members of Leptoconopinae NoeÁ and the subfamily Austroconopinae Borkent, Wirth and Dyce is a new synonym of Leptocono- pinae. Extant and fossil distributional records suggest that Austroconops was displaced from its previously broad distribution by the emergence of Latreille. Larval feeding on microorganisms is plesiotypic within the Ceratopogonidae and , and living in small aquatic habitats is plesiotypic within each family of the Culicomorpha. Outgroup com- parisons further suggest that diurnal feeding by adult females is plesiotypic within the Cera- topogonidae and Chironomoidea.

INTRODUCTION bers of Leptoconops, species of Austrocon- ops have a number of plesiomorphic features, Perhaps only fellow entomologists could allowing for an enhanced interpretation of appreciate the thrill of the following sado- character states within the family. Simply masochistic scenario: the ®rst author and his put, Austroconops is a group of primitive wife standing still on the edge of a golf ``living fossils''. Further to this, knowing that course in Yanchep National Park, just north the Early Cretaceous species of Austrocon- of Perth in Western Australia, being badly ops were morphologically similar to the ex- bitten on the face by hundreds of biting tant species suggested that study of the bi- midges, with exclamations of joy and won- ology of the living species would provide der. We realized that we were being attacked special insight into the adaptations of Cera- by a species considered to be an extremely topogonidae existing 121 million years ago. rare ``living fossil'', a member of a genus Of particular interest and importance was the which was surviving only in this small cor- discovery of the eggs and larvae of the two ner of the globe but which was once a wide- extant species, as well as the pupa of one of spread and diverse lineage during the Early these. One of the reasons that nematocerous Cretaceous, 121 million years ago. Previous- Diptera are such excellent candidates for ly known from only a limited series of adult phylogenetic studies is that the different life specimens of a single species and a moderate stages show a remarkable degree of morpho- number of fossils, our study of this genus logical divergence and therefore provide in- uncovered a wealth of new information: the dependent sources of character states to test discovery of the previously unknown eggs, phylogenetic hypotheses. larvae, and pupa; the presence of a second When Austroconops and its single species extant species (including its immatures); and A. mcmillani was ®rst described by Wirth observations of female biting behavior, male and Lee (1958) only 39 females were known swarming, and more. from four localities in the vicinity of Perth, There are good reasons why we were ex- Western Australia. These authors noted that cited to pursue these small beasts, and we the species seemed closest to Culicoides La- committed ourselves to an extensive expe- trielle, well within the Ceratopogonidae, but dition to Western Australia to seek them out. mentioned that it shared some features with In recent years cladistic analyses and an the early lineage Leptoconops. They were abundance of fossil material in amber of dif- uncertain as to which subfamily the genus ferent ages have resulted in an increasingly belonged. The only information on the spe- detailed understanding of the phylogenetic cies' biology was that females were recorded relationships of the biting midges. The genus as biting humans, particularly on the eyelids. Austroconops Wirth and Lee plays a pivotal Then, in 1985, W.W. Wirth and A.L. Dyce role in this analysis. Together with Leptocon- collected the ®rst males of the species, and ops Skuse, these two ancient genera repre- study of these three specimens (plus an ad- sent the earliest extant lineage in the family. ditional 10 females) led to a cladistic analysis However, unlike the rather modi®ed mem- of the early lineages of the family, demon- 2004 BORKENT AND CRAIG: BITING MIDGES FROM AUSTRALIA 3 strating that Austroconops belonged near the Subsequent care was as described below for very base of the phylogeny of the Cerato- A. mcmillani. Females of A. mcmillani were pogonidae (Borkent et al., 1987). Since then, allowed to attack the face at site A (see ®g. six fossil species have been described from 22C), complete their feeding, and then were Early to Late Cretaceous amber (from Leb- captured either by placing a small vial over anon, France, and Siberia), providing, within the female until she ®nished feeding or by a cladistic analysis, further support that the sticking one's head in an net and gent- genus indeed represents an early lineage of ly aspirating the fully fed females after they biting (Borkent, 2000a). Within the left the skin surface and ¯ew into the net. Ceratopogonidae, there is a striking correla- These captured females of A. mcmillani were tion between cladistic patterns and the dis- then cooled brie¯y in a refrigerator and when tribution of fossils in different ages of amber, torpid, quickly placed in six groups of 7±12 with increasingly older fossils representing females each and put into separate 2- or 3- increasingly earlier lineages (Borkent, dram vials containing about 1 cm of wet 2000a). Our study here further re®nes the mud/detritus taken from the partially inun- phylogenetic position of Austroconops, inter- dated margin at the south end of Loch prets characters states of the newly discov- McNess (®g. 22C; site B). Females of both ered immatures, analyzes the implications for species received a small smear of honey on our interpretation of features of other Cera- the inside surface of the vial at about mid- topogonidae, and provides an interpretation height (none were seen to feed) and each vial of historical zoogeographical and bionomic was stoppered with a cotton ball. A drop of features of early lineages of Ceratopogonidae water was added periodically to the mud and other Culicomorpha. when it began to dry (so the mud remained quite wet). The vials were kept at more-or- MATERIALS AND METHODS less ambient temperature in a small cooler that was always kept in the shade. Females The collecting trip to Western Australia of A. mcmillani were transported from Aus- was undertaken by the ®rst author and his tralia to Canada with the addition of a large wife with the purpose of discovering further bottle of very warm water to the cooler to material of Austroconops. Sampling was pri- keep adults from becoming torpid and stick- marily with a sweep net in a wide array of ing to the mud surface. The eggs of A. an- wet or aquatic habitats from October 12 to nettae, which had been laid while in Austra- November 23, 2001, covering the area from lia, plus ®rst instar larvae were transported Yanchep N.P., about 47 km north of Perth, to Canada in the same cooler. In Canada the south to Augusta, east to Albany, returning eggs and larvae of A. annettae and the fe- through Darkan to Augusta and then heading male adults, eggs and larvae of A. mcmillani north to Yanchep N.P. once again. Mud sam- were kept at about 23±28ЊC during the day ples were taken from wet habitats near the and about 18ЊC at night. Most of the females sites of where adult Austroconops were col- of A. mcmillani that were transported died en lected and immatures searched for (unsuc- route, probably because of excessive shaking cessfully) either manually or by ¯oating de- of the vials during air travel; of 58 original bris and substrate using a saturated sugar so- females, only 12 survived the trip to Canada. lution. Female adult A. mcmillani were col- Of these, at least one of three females laid lected with a net and aspirator by Len eggs. Zamudio during December, 2001±January, Eggs were allowed to develop in the vials, 2002 and October, 2002±February, 2003 and after all had hatched, the mud contents while the midges were attacking humans in with the larvae of each species was trans- Yanchep National Park. ferred to small (50 ϫ 7 mm) plastic petri To obtain eggs of A. annettae, an adult fe- dishes with lids. The larvae could be easily male was captured by sweeping and placed observed moving or lying against the bottom directly into a vial with mud/detritus (about of the dish by inverting the clear dish under 1 cm deep) taken from the spot where the the dissecting microscope. Vials and dishes adults of this species had been collected. were examined at least twice a day for the 4 AMERICAN MUSEUM NOVITATES NO. 3449 great majority of larval and pupal develop- were examined under both dissecting and ment. compound microscopes by placing them in a The mud/detritus substrate initially con- well slide in which they were studied while tained some small, unidenti®ed microorgan- warm and after they were cooled by ice. Cold isms, but nearly all died shortly after obtain- larvae were examined for mouthpart and pro- ing the samples. When it became clear that leg movement. the ®rst-instar larvae were not feeding, be- Adult behavior of A. mcmillani was ob- tween hatching and December 8, 2001, the served at the south end of Loch McNess (®g. following items were added as potential food 22C; site B) and on fairway 2 of the golf at different times to the Petri dish with larvae course (®g. 22C; site A) at Yanchep National of A. mcmillaniÐEuglena sp., yeast, a fresh- Park. Biting times by female A. mcmillani ly killed ostracod and Chaoborus Lichten- were determined by observing a female land, stein , several species of unidenti®ed waiting for initiation of feeding, and then green algae and cyanobacteria (from local carefully inverting a vial over the female un- ®sh tanks and ponds), dried blue-green algae til it ceased feeding, when it invariably ¯ew (Spirulina) soaked in water, and the follow- up into the vial. ing cultured cyanobacteria and green algae: Specimens were examined, measured, and Anabaena ¯osaquae, Oscillatoria sp., and drawn using a Wild M3 dissecting micro- Selenastrum capricornutum. An unidenti®ed scope and a Zeiss Jenaval compound micro- species of rotifer was added between Decem- scope. Larval head capsule length was mea- ber 31, 2001 and January 31, 2002. A small sured from the posterolateral margin to the amount of water was added to agar plates anterolateral margin of the head capsule with a culture of the nematode Caenorhab- (near the base of the mandible). A male and ditis elegans and the bacterium Escherichia female of both species were cleared and coli, swirled around, and 2±5 drops of the compared in temporary glycerine slide water with C. elegans were added every 2± 3 days to the mud/detritus in the Petri dishes mounts. These and other adults, as well as with A. mcmillani and A. annettae starting on most eggs and larvae were mounted on mi- December 31. These nematodes died within croscope slides using the technique described 12±24 hours. Beginning on December 17, 2± by Borkent and Bissett (1990). Larvae to be 4 drops of an infusion made from fresh goat studied with the scanning electron micro- feces and water (®rst made on December 15) scope were killed with hot water. Some eggs were added every 2±3 days to both dishes; and larvae were critical point dried, sputter- these drops regularly included a small clot of coated at 30 angstroms with chrome, and feces. Water was added to the infusion to re- studied with a Jeol JSM 6301 FXV scanning place loss through evaporation (the infusion electron microscope. was not sealed), and a fresh infusion was Terms for structures follows those used in made on August 9, 2002. the Manual of Nearctic Diptera (McAlpine, Five ®rst-instar larvae of A. mcmillani 1981). More speci®c larval and pupal terms were added to an agar plate with a culture of follow Lawson (1951). The kangaroos re- C. elegans and E. coli on December 14 and ferred to in the text are all western grey kan- were seen feeding on clumps of E. coli, but garoos (Macropus fuliginosus). The name all died within a few days. At least two of Austroconops annettae is attributed solely to these larvae had their guts partially ®lled the ®rst author. with white material, almost certainly E. coli. Additional material was borrowed from On June 28, 2002, three third-instar A. the following museums: mcmillani were placed in a separate dish with ANIC Australian National Insect Collection, some of the original mud/detritus, put into a Division of Entomology, CSIRO, P.O. dark cold room for four days at 13ЊC, and Box 1700, Canberra City, A.C.T. 2601, then placed in a refrigerator at 5±8ЊC. By Australia August 1 all larvae had died and were pre- BMNH Nigel Wyatt, Department of Entomol- served in 70% ethanol. ogy, Natural History Museum, London, Some living larvae of different instars SW7 5BD, United Kingdom 2004 BORKENT AND CRAIG: BITING MIDGES FROM AUSTRALIA 5

BPBM Department of Entomology, Bernice P. mere 12 with or without subbasal constric- Bishop Museum, 1525 Bernice Street, tion, ¯agellomere 13 with subbasal constric- P.O. Box 19000A, Honolulu, HI, 96817 tion, ¯agellomere 1 with two patches of short 0916 sensilla trichodea, without sensilla coelocon- CNCI Canadian National Collection of In- ica. Mouthparts moderately short to moder- sects, Eastern Cereal and Oilseed Re- search Centre, Agriculture Canada, ately long. Mandible elongate (ending near K.W. Neatby Building, Ottawa, Ontar- apex of labrum), with several slender termi- io, K1A 0C6, Canada nal spicules. Lacinia elongate, simple. Palpus USNM United States National Entomological (®g. 1C, D) with 4±5 segments, segment 3 Collection, Department of Entomology, ovoid to elongate, slender, with capitate sen- U.S. National Museum of Natural His- silla scattered on mesal surface or perhaps in tory, Washington, DC 20560 pit, at least extant species with membranous WAMP Department of Entomology, Western area between segment 3 and 4 ϩ 5. Thorax: Australian Museum, Francis Street, With three anterior pronotal apodemes. Scu- Perth, WA, 6000, Australia tum with scattered elongate setae. Scutellum rounded or angular in dorsal view. Anapleur- RESULTS al suture elongate. Wing (®g. 1K): Without Austroconops Wirth and Lee macrotrichia, ®ne microtrichia present on all membrane. Alula with fringe of macrotrichia. Austroconops Wirth and Lee, 1958: 337. Type Costa extending to or beyond apex of R3. species: Austroconops mcmillani Wirth and Both radial cells present. r-m parallel to R . Lee, by original designation. 1 M bifurcating distal to r-m. Legs: Femora, DIAGNOSIS: Male. The only extant or fossil tibiae slender. Legs lacking armature, except Ceratopogonidae with ¯agellomere 13 (®g. in some species with stout setae on some or 1A, B) with a subbasal constriction, with two all of ®rst tarsomeres, some with pair of thick well-developed radial cells, and r-m parallel setae on apex of tarsomeres 1±4. Tarsal ratio to R1 (®g. 1K). Female. The only extant or (Ta1/Ta2) of foreleg/hindleg ϭ 1.4±1.9. Fore- fossil Ceratopogonidae with two well-devel- and midleg trochanter without pair of thick oped, clearly open, radial cells and r-m par- setae. Midleg tibia with or without apical allel to R1 (®g. 1L). Egg. Only - spur. Hindtibia apex with two rows of spines. idae with egg (®g. 2A) markedly elongate Hindleg ®rst tarsomere with or without thick and remaining pale throughout larval devel- basal spine, with scattered setae (®g. 1M, N). opment. Larva (all instars). The only Cer- Claws on fore-, mid-, hindleg equal in size, atopogonidae with markedly elongate anten- both equal on each leg, each claw simple or na bearing an elongate blade and a progna- toothed, apically bi®d (possibly simple in thous head (®gs. 2H, 3C, 12A, B). Pupa. The Lebanese amber fossils). Slender empodium. only Ceratopogonidae with the hindleg Genitalia: Apicolateral process absent to sheath not exposed from under lateral margin well developed. Gonocoxite short to moder- of wing sheath and with abdominal segments ately elongate. Gonostylus variable, apical 2±8 with bifurcate setae (®g. 4A). spine present or absent. Parameres fused me- DESCRIPTION: Live adults of both sexes. dially (known only in extant species). Ae- With subcutaneous blue-green tissue (likely deagus short, setose lobe with ventral plate fat body) externally evident in all areas (known only in extant species). (head, thorax, abdomen) with membrane or Female adult. Head: Ommatidia narrowly thin cuticle (including tip of halter). Wings separated dorsomedially, with single vertex completely overlapping at rest. Male with seta. Antenna with 13 separate ¯agellomeres, permanently erect antennal setae. Male ¯agellomeres gradually increasing in length adult. Head: Ommatidia narrowly separated from ¯agellomeres 2±13 or with ¯agello- dorsomedially, with single vertex seta. An- meres 9±13 more elongate than preceding tenna (®g. 1A, B) with well-developed plume ¯agellomeres, ¯agellomere 1 with two of permanently erect setae, 13 separate ¯a- groups of short sensilla trichodea, without gellomeres, ¯agellomeres 12±13 more elon- sensilla coeloconica. Mouthparts moderately gate than preceding ¯agellomeres, ¯agello- elongate, further details not visible in fossils. 6 AMERICAN MUSEUM NOVITATES NO. 3449

Fig. 1. Structures of adult Austroconops. A. Male antenna of A. mcmillani. B. Male antenna of A. annettae. C. Right palpus of male A. mcmillani. D. Right palpus of male A. annettae. E. Right palpus of female A. mcmillani. F. Right palpus of female A. mcmillani. G. Right mandible of A. mcmillani in anterior view. H. Right mandible of A. annettae in anterior view. I. Hindleg claws of A. mcmillani. J. Hindleg claws of A. annettae. K. Right wing of male A. annettae. L. Right wing of female A. annettae. M. Midleg of male A. mcmillani. N. Midleg of male A. annettae. Scale bars: A, B ϭ 0.1 mm; C±F ϭ 0.05 mm; G±J ϭ 0.01 mm; K, L ϭ 0.1 mm; M, N ϭ 0.05 mm. 2004 BORKENT AND CRAIG: BITING MIDGES FROM AUSTRALIA 7

Fig. 2. Structures of immature Austroconops. A. Egg of A. mcmillani. B. Detail of tubercles on surface of egg of A. mcmillani in lateral view. C. First-instar larva of A. annettae in lateral view. D. First-instar larva of A. mcmillani in lateral view. E. Second-instar larva of A. mcmillani in lateral view. F. Third-instar larva of A. mcmillani in lateral view. G. Fourth-instar larva of A. mcmillani in lateral view. H. First-instar larval head capsule of A. mcmillani in dorsal view. I. First-instar larval right mandible and apodeme of A. mcmillani in dorsal view. J. First-instar larval pharyngeal complex of A. mcmillani from ®gure K. K. First-instar larval head capsule of A. mcmillani in ventral view. Scale bars: A, C±G ϭ 0.1 mm; B, H±K ϭ 0.01 mm. 8 AMERICAN MUSEUM NOVITATES NO. 3449

Fig. 3. Structures of larvae of Austroconops. A. Fourth-instar larval head capsule of A. mcmillani in anterior view; antennae and maxillae not shown. B. Third-instar larval posterior portion of segment 9ofA. mcmillani in lateral view. C. Fourth-instar larval head capsule of A. annettae in dorsal view. D. Fourth-instar larval epipharyngeal bar and premandibles of A. annettae in dorsal view. E. Fourth-instar larval left mandible and apodeme of A. annettae in dorsal view. F. Fourth-instar larval pharyngeal complex of A. annettae. G. Fourth-instar larval head capsule of A. annettae in ventral view. H. Fourth- instar larval segment 9 of A. mcmillani in dorsal and ventral view. Scale bars: A±G ϭ 0.05 mm; H ϭ 0.1 mm; D±F are from ®gures C and G. 2004 BORKENT AND CRAIG: BITING MIDGES FROM AUSTRALIA 9

Fig. 4. Structures of immatures of Austroconops mcmillani. A. Habitus of pupa in lateral view; am ϭ anteromedial, dl ϭ dorsolaterals, ad ϭ anterodorsal, vl ϭ ventrolaterals, d ϭ dorsals. B. Third-instar larval prothoracic segment in lateral view. C. Pupal head in ventral view; vm ϭ ventromedial, vl ϭ ventrolaterals. D. Pupal operculum. E. Pupal right anteromedial seta in anteroventral view. F. Pupal dorsolateral setae in dorsal view. Scale bars: A, C±E ϭ 0.1 mm; B, F ϭ 0.05 mm. 10 AMERICAN MUSEUM NOVITATES NO. 3449

Fig. 5. Structures of the pupa of Austroconops mcmillani. A. Right portion of cephalothorax in dorsal view; am ϭ anteromedial, dm ϭ dorsomedial, dl ϭ dorsolaterals, ad ϭ anterodorsals, d ϭ dorsals. B. Right respiratory organ in dorsal view. C. Right respiratory organ in lateral view. D. Seta dpm i in dorsal view. E. Seta vn ii in ventral view. F. Leg and wing sheaths in ventral view. G. Abdominal segment 4 in dorsal and ventral view. Scale bars: A, F, G ϭ 0.1 mm; B±E ϭ 0.05 mm. 2004 BORKENT AND CRAIG: BITING MIDGES FROM AUSTRALIA 11

Fig. 6. Structures of the pupa of Austroconops mcmillani. A. Posterior portion of scutum, metatho- rax, and ®rst abdominal tergite in dorsal view. B. Segment 9 in dorsal and ventral view. Scale bars ϭ 0.1 mm.

Mandible (®g. 1G, H) and laciniae with ®ne Scutum with scattered elongate setae. Scu- teeth in extant species. Palpus (®g. 1E, F, tellum angular in dorsal view. Anapleural su- 23D) with 4±5 segments, segment 3 ovoid to ture elongate. Wing (®g. 1L): Without ma- elongate, with capitate sensilla scattered on crotrichia, ®ne microtrichia present on all mesal surface, at least extant species with membrane. Alula with or without fringe of membranous area between segments 3 and 4 macrotrichia. Costa extending to or beyond

ϩ 5. Thorax: With three anterior pronotal apex of R3. Both radial cells present. M bi- apodemes (known only in extant species). furcating distal to r-m. r-m parallel to R1. 12 AMERICAN MUSEUM NOVITATES NO. 3449

Legs: Femora, tibiae slender. Legs lacking ar- slender, with apical pit with small peg; sen- mature except in some with pair of stout se- sillum 2 moderately elongate; sensillum 3 tae on apex of tarsomeres 1±4 of all legs. TR short, bilobed; sensillum 4 moderately elon- of foreleg/hindleg ϭ 1.7±2.2. Fore- and mid- gate, with pores. Labrum (®gs. 3A, 15A, B, leg trochanter without pair of thick setae. 17C, 19) wide, with well developed torma, Midleg tibia with or without apical spur. with well developed, dark, articulating, api- Hindtibia apex with two rows of spines. Hin- cally bent premandible; torma abutting lat- dleg ®rst tarsomere with or without thick eral margin of stout, black, transverse epi- basal spine, with scattered slender or stout pharyngeal bar; epipharyngeal bar forming setae. Claws on fore-, mid-, hindleg equal in large, ventrally directed, internal, median size, both equal on each leg, each claw sim- lobe, posterodorsally directed bilobed apo- ple or toothed (®g. 1I, J). Empodium slender. deme and lateral apodeme abutting torma; la- Genitalia: Two large, one markedly smaller brum with 10 sensilla: sensilla 1±4 an anter- spermathecae. Sternite 9 continuous medial- odorsal group, with sensillum 1 a peg in pit, ly. Segment 10 with pair of setae. Cerci short sensillum 2 a styloconicum on cuticular pro- to moderately elongate. jection, sensilla 3, 4 lobe-shaped with 4 lon- Egg (®g. 2A): Very light yellowish brown, ger than 3, sensillum 5 an elongate seta, sen- appearing nearly white during development silla 6, 7 basally stout basiconica, sensilla 8, of larva. Elongate and slender. With 9±10 9 laterally placed, sensillum 8 short seta, sen- longitudinal rows of short, minute tubercles, sillum 9 a short peg, sensillum 10 a stout each tubercle slender at base, expanded dis- seta; ventral margin with well developed sco- tally (®g. 2B). Anterior end more rounded pae, 18±21 more-or-less uniform teeth (three than posterior end. Eggshell opening a single lateral teeth more stout) in undivided row. dorsal slit along most of length. Mandible (®gs. 2I, 3E, 10A) curved, apical All larval instars: Head capsule (®gs. 2H, half tapering to sharp, darkly pigmented, K, 3A, C, G, 7A, B, 8A, B, 12A, B, 13A, point, with dorsal and ventral grooves; with B): Nearly square in dorsoventrally viewed large to very small lobe (not visible in some outline; light brown, with only pigmentation specimens) on inner surface; with subbasal present: dark epipharyngeal bar, premandi- seta and minute peg in pit on outer margin; ble, apex of mandible; eye in later instars with well developed apodeme attached to (some seconds, all thirds, fourths) a single, dorsal margin of mandible, extending into roughly circular to oval spot. Ecdysial suture posterior half of head capsule. Maxilla (®gs. extended anteriorly slightly beyond level of 10B, 16B, 17D) well developed, wide; pal- sensillum s, possibly longer but dif®cult to pus elongate; large lobelike, apically tapering discern; area of thinner cuticle broad in area plate dorsal to base of palpus; with postero- of sensilla j (anterior one), p, r. Ventral suture medial elongate seta; palpus with 4 apical or not present. Setae all simple (not bifurcate), subapical sensilla, two lateral sensilla each arrangement as in ®gures 2H, K, 3C, G, 7A, with elongate slender nib; group of 7 thick B, 8A, 12A, B, 13A, B; following sensilla sensilla lateral to palpus: sensillum 1 extend- present: x (both setae present), t (or possibly ing to about half length of palpus; sensillum an anteriorly placed q), s, p (two long setae, 2 with rounded apex; sensillum 3 short, aris- posterior one expanded distally), r (a short ing near base of sensillum 4; sensillum 4 peg), j (anterior one a short peg on head cap- with elongate apical nib; sensillum 5 on elon- sule; other a more elongate seta on anterior gate cuticular extension; sensillum 6 with margin of cervix), o (both an equally long rounded apex, ribbed; sensillum 7 with seta), w, u, m, v, and y. Antenna (®gs. 9A, rounded apex; lateral seta elongate. Pharyn- B, 14A, B) elongate, with three segments; geal complex (®gs. 2J, 3F) well developed, with well developed, short, basal segment, epipharynx with two lateral arms, apparently segments 2±3 short, slender, segment 3 ta- lacking combs (present but dif®cult to dis- pering to slender, elongate apex; ®rst seg- cern in some other Ceratopogonidae). Hy- ment otherwise bearing greatly elongate, popharynx with lateral arms articulating with multiporous blade and ®ve other sensilla: ac- lateral apices of lateral arms of epipharynx. cessory blade, sensilla 1±4; sensillum 1 long, Thorax, abdomen: Cuticle unpigmented, 2004 BORKENT AND CRAIG: BITING MIDGES FROM AUSTRALIA 13

Fig. 7. Structures of the ®rst-instar larva of Austroconops mcmillani. A. Head capsule in dorsal view. B. Head capsule and anterior proleg in lateral view. Scale bars ϭ 10 ␮m. 14 AMERICAN MUSEUM NOVITATES NO. 3449

Fig. 8. Structures of the ®rst-instar larva of Austroconops mcmillani. A. Head capsule and anterior proleg in ventral view. B. Head capsule in anterodorsal view. Scale bars ϭ 10 ␮m. 2004 BORKENT AND CRAIG: BITING MIDGES FROM AUSTRALIA 15

Fig. 9. Right antenna of the ®rst-instar larva of Austroconops mcmillani;sϭ sensillum. A. In dorsal view. B. In mesal view. Scale bars: A ϭ 2 ␮m, B ϭ 1 ␮m. 16 AMERICAN MUSEUM NOVITATES NO. 3449

Fig. 10. Structures of the ®rst-instar larva of Austroconops mcmillani;sϭ sensillum. A. Right mandible and right portion of labrum in anterodorsal view. B. Right maxilla in anterodorsal view. Scale bars ϭ 2 ␮m. 2004 BORKENT AND CRAIG: BITING MIDGES FROM AUSTRALIA 17

Fig. 11. Abdominal segment 9 of the ®rst-instar larva of Austroconops mcmillani. A. With partially extruded proleg and anal papillae, in posterodorsal view. B. In lateral view. Scale bars ϭ 10 ␮m. 18 AMERICAN MUSEUM NOVITATES NO. 3449

Fig. 12. Structures of the fourth-instar larva of Austroconops mcmillani. A. Head capsule in dorsal view. B. Head capsule and anterior proleg in lateral view. Scale bars ϭ 20 ␮m. 2004 BORKENT AND CRAIG: BITING MIDGES FROM AUSTRALIA 19

Fig. 13. Structures of the fourth-instar larva of Austroconops mcmillani. A. Head capsule and an- terior proleg in ventral view. B. Head capsule in anterolateral view. Scale bars ϭ 20 ␮m. 20 AMERICAN MUSEUM NOVITATES NO. 3449

Fig. 14. Antennae of the fourth-instar larva of Austroconops mcmillani;sϭ sensillum. A. Right antenna in dorsal view (see ®g. 12A for less magni®ed view). B. Left antenna in distal view. Scale bars: A ϭ 5 ␮m, B ϭ 1 ␮m. 2004 BORKENT AND CRAIG: BITING MIDGES FROM AUSTRALIA 21

Fig. 15. Structures of the fourth-instar larva of Austroconops mcmillani;sϭ sensillum. A. Right portion of labrum in dorsoanterolateral view. B. Labrum, right antenna, and right mandible in antero- lateral view. Scale bars: A ϭ 2 ␮m, B ϭ 5 ␮m. 22 AMERICAN MUSEUM NOVITATES NO. 3449

Fig. 16. Structures of the fourth-instar larva of Austroconops mcmillani;sϭ sensillum. A. Anterior portion of head capsule in ventral view. B. Right maxilla in anterolateral view (see Fig. 15B for less magni®ed view of base of maxilla). Scale bars: A ϭ 10 ␮m, B ϭ 2 ␮m. 2004 BORKENT AND CRAIG: BITING MIDGES FROM AUSTRALIA 23

Fig. 17. Structures of larvae of Austroconops mcmillani;sϭ sensillum. A. Fourth-instar larval anterior proleg in anterolateral view. B. Fourth-instar larval posterior portion of abdominal segment 9 in dorsolateral view. C. Third-instar larval labrum in anterior view. D. Third-instar larval left maxilla in anterior view. Scale bars: A, B ϭ 10 ␮m, C ϭ 2 ␮m, D ϭ 1 ␮m.

transparent, thin. Prothorax secondarily di- proximated) setae: dorsal setae o, i, l1,d,l2, vided, with well developed cervix; with elon- l3,l4, ventral setae i, o, l1,l2, v; posterior pro- gate proleg (®gs. 2C±G, 4B, 7B, 12B, 17A, leg (®gs. 2C±G, 3B, 11A, B, 17B) a single 18), with apical hooks, 5±6 (per half) ante- posterior structure with about 15±20 well de- rior terminal hooks elongate, posterior hooks veloped hooks, dorsal hooks with broader short; proleg capable in life of being with- bases than ventral, more slender hooks; ven- drawn into prothorax (posterior to cervix). tral hooks with spicules; proleg capable of Segment nine (®gs. 3B, H, 11A, B) with being extruded or withdrawn into body cav- well-separated (with bases not closely ap- ity. Four anal papillae, each apically bifur- 24 AMERICAN MUSEUM NOVITATES NO. 3449

Fig. 18. Head capsule and anterior proleg of the third-instar larva of Austroconops annettae in lateral view. Scale bars ϭ 20 ␮m.

cate. Midgut white, with annulations (ob- B) with at least dorsal setae o, i, d, l1,12,13, scured by fat body in some third- and fourth- ventral setae o, i present; others not visible instar larvae); anterior margin situated at but may be present; dorsal seta o notably midlength of fourth true abdominal segment thicker, longer than other setae on segment. (at anterior margin of apparent segment 8 in Hemolymph unpigmented. larvae with secondarily divided segments). Second-instar larva: Head capsule: With With two Malpighian tubules. or without eyespot. Mandible likely (not First-instar larva: Head capsule (®gs. clearly visible) with moderately sized trian- 7A, B, 8A): With dorsal, darkly pigmented gular tooth on inner margin. Hypostoma with egg burster (®gs. 2H, 7A, B, 8B). Without well developed row of teeth. Abdomen: With eyespot. Mandible (®g. 2I) with large trian- segments 1±8 secondarily divided (so abdo- gular tooth on inner margin (not evident in men appears to have 17 segments) (®g. 2E). some specimens). Hypostoma lacking teeth Abdominal segment 9 with uncertain number (®gs. 2K, 8A) Abdomen: With or without of setae; dorsal seta o at least slightly thicker segments 1±8 secondarily divided (so abdo- than other setae on segment. Hemolymph un- men appears to have either 9 or 17 segments) pigmented or very pale pink. (®g. 2C, D). Abdominal segment 9 (®g. 11A, Third-instar larva: Head capsule: With 2004 BORKENT AND CRAIG: BITING MIDGES FROM AUSTRALIA 25

Fig. 19. Head capsule of the third-instar larva of Austroconops annettae in anterior view; s ϭ sensillum. Scale bar ϭ 5 ␮m. eyespot. Mandible with small bump on inner have 17 segments) (®g. 2G). Abdominal seg- margin. Hypostoma (®g. 19) with well de- ment 9 with 7 dorsal, 5 ventral setae distrib- veloped row of teeth, central tooth largest. uted as in ®gures 3H, 17B; dorsal seta o Abdomen: With segments 1±8 secondarily di- equal in diameter to other long setae on seg- vided (so abdomen appears to have 17 seg- ment. Hemolymph pink or reddish. Fat body ments) (®g. 2F). Abdominal segment 9 with present. 7 dorsal, 5 ventral setae distributed as in ®g- Pupa: Only pupa known is of A. mcmil- ures 3H, 17B; dorsal seta o at least slightly lani, described below. thicker than other setae on segment. Hemo- lymph unpigmented or pink. Some fat body DISTRIBUTION AND BIONOMICS visible in more mature larvae. Fourth-instar larva: Head capsule: With The only two extant species are restricted eyespot. Mandible (®g. 3E) with small bump to southwestern Australia (®g. 22B), but Cre- on inner margin. Hypostoma (®gs. 3G, 16A) taceous fossils are known from France, with well developed row of teeth, central Spain, Siberia, Lebanon, and Myanmar (Bor- tooth largest. Abdomen: With segments 1±8 kent, 2000a; Szadziewski, in press; Szad- secondarily divided (so abdomen appears to ziewski and Arillo, 2001), proving that the 26 AMERICAN MUSEUM NOVITATES NO. 3449 genus was once much more broadly distrib- mcmillani may have indicated that the then uted. unknown larvae were feeding on algae be- The only species in which females have cause this coloration has been observed in been observed to bite are those of A. mcmil- some Ceratopogonidae such as some Culi- lani, which feed on kangaroos and humans. coides species of the schulzei species group However, the ®nely serrate mandible and re- and some Dasyhelea Kieffer species which trorse lacinial teeth of the adult females of feed on algae as larvae. Our observations A. annettae strongly indicate that these too here show that the relationship between algal feed on (Borkent, 1995: 129± feeding and adult color is not substantiated 132). The morphology of the claws of the for at least A. mcmillani as larvae matured to female of A. annettae additionally may in- adulthood without feeding on algae. Austro- dicate that these feed on birds (see discussion conops annettae was also successfully reared below under that species). The mouthparts of to the fourth-instar without algae. fossil Austroconops are not visible (Borkent, 2000a), and therefore it is uncertain what TAXONOMIC DISCUSSION they fed upon. The presence, however, of blood-feeding in A. mcmillani and We consider the elongate, pale egg of Aus- the phylogenetic position of the genus as an troconops species unique within the family. early lineage within the family where verte- Eggs laid by other Ceratopogonidae are ini- brate blood-feeding is plesiotypic strongly tially pale but soon turn dark. However, of suggest that all extinct species fed on verte- 103 extant genera, eggs have been described brates (Borkent, 1995, 2000a). Details of for only 18 genera and of those, 4 genera are male swarming in A. mcmillani are given be- known only as eggs described from within low. the female abdomen (so their ®nal color can- First-instar larvae of A. annettae and A. not be determined). mcmillani were both successfully reared to We identi®ed the four larval instars of fourth-instar larvae (and A. mcmillani to the both species based on the following evi- pupal stage) in very wet soil, with regular dence. First-instar larvae are easily identi®ed (generally every second day) additions of by their darkly pigmented egg burster (®gs. nematodes and a fecal infusion. All instars of 2H, 7A, B, 8B). Because of the small number the aquatic larvae were clearly attracted to of measured second and fourth-instars of fresh drops of fecal infusion as shown by the reared Austroconops larvae it was initially concentrations of larvae directly under these dif®cult to identify the instars 2±4. The drops and they were rarely seen to feed on fourth-instar of A. mcmillani was con®dently nematodes. This phenomenon, and the pres- identi®ed because one of these molted to the ence of well developed, ®nely toothed scopae pupal stage. Although the head capsule of (®gs. 3A, 19) suggest that they are likely as- this individual was not retrieved from the sociated with feces or concentrated decom- mud and therefore not measured, observa- posing vegetation in nature (producing an tions before it pupated showed that it was abundance of microorganisms). Some sec- clearly close to, or within, the range of mea- ond, nearly all third, and all fourth-instar lar- surements reported here. Head capsule length vae had pink or red hemolymph, indicating increments of all instars followed Dyar's the presence of hemoglobin, which addition- Law, increasing by a factor of 1.32, 1.31, and ally suggests that they may be associated 1.34 for A. mcmillani and 1.33, 1.32, and with an oxygen de®cient wet habitat. Larvae 1.25 for A. annettae. The latter value, the cannot swim but use a combination of their factor of change for third to fourth-instar, anterior and posterior prolegs and, especially may be due to the small sample size of in later instars, a relatively slow serpentine fourth-instar A. annettae or perhaps to less body motion to move through wet substrate. than optimal feeding conditions (and hence Further details of behavior and habitat are smaller individuals). These values are similar provided below for each species. to those reported for species of Culicoides, Borkent et al. (1987) suggested that the which are about 1.4 (Kettle and Lawson, blue-green pigmentation of live adult A. 1952; Kettle and Elson, 1975). 2004 BORKENT AND CRAIG: BITING MIDGES FROM AUSTRALIA 27

There are signi®cant statistical differences these are likely homologous. in egg characteristics (table 3) and larval S1, just dorsal to the labral lamellae, proba- head capsule lengths (table 4) of the different bly is homologous to our S10, and the scopae instars between A. mcmillani and A. annet- present in Austroconops and Ceratopogoni- tae. Considering, however, that these imma- nae are probably homologous to the labral tures were obtained from eggs laid by a very lamellae (Wiederholm, 1983). Further de- few females, and that the larvae were reared tailed comparisons require further study of under laboratory conditions, the size differ- these structures in numerous taxa (both SEM ences noted here may be artifacts. Otherwise, and study of nerves). eggs and larvae of the two species could not Borkent et al. (1987) reported an ``M- be distinguished from one another. shaped apodeme'' in the female genitalia of Larvae of Austroconops, when compared A. mcmillani. In further study we are puzzled to other Ceratopogonidae, are missing head as to the nature of this structure, which varies capsule sensilla z, k (sensory pit), and q. in both A. mcmillani and A. annettae from a Also, there are two additional dorsolateral se- faint M-shaped sclerotization to the presence tae on larval abdominal segment 9 (®g. 3H), of a pair of laterally positioned, pigmented here labeled as l3 and l4, which have not been sclerites. Some other Ceratopogonidae also reported in other Ceratopogonidae. have sclerotized structures anterior to the The larval antennal sensilla of Ceratopo- fused or separated sternite 9 (e.g., some Lep- gonidae have never been described in suf®- toconops, some Culicoides, cient detail to determine most homologies Kieffer, and others). Careful histological within and outside the family. Larvae of Aus- study is needed to better interpret this fea- troconops species have a well-de®ned basal ture. segment bearing 6 apical sensilla and a fur- Cladistic analysis below indicates that, ther second and third segment (®gs. 9A, B, among extant taxa, Austroconops is the sister 14A, B, 20A). Based on position, relatively group of Leptoconops and that together they large size, and a uniformly porous surface, form the sister group to all remaining extant the most elongate, porous sensillum is ho- Ceratopogonidae. Eight species of Austro- mologous to the ``large lobe'' present in Cu- conops are now known: two of these are ex- licoides (Murphree and Mullen, 1991) and tant and six are Cretaceous fossils: most other Ceratopogoninae (Borkent and Bissett, 1990; Borkent and Craig, 1999). A. annettae, n.sp. Borkent, this publication. Aus- Based on position and some details of struc- tralia (Western Australia) ture, the following sensilla are likely homol- A. borkenti Szadziewski and SchluÈter, 1992: 78. ogous to sensilla in Chironomidae: most France. Upper Cretaceous elongate, porous sensillum ϭ blade; mesal A. fossilis Szadziewski, 1996: 38. Lebanon. Low- short sensillum ϭ accessory blade; short bi- er Cretaceous lobed sensillum labeled sensillum 3 here ϭ A. gladius Borkent, 2000a: 378. Lebanon. Lower fused style and peg sensillum. The remaining Cretaceous. three short sensilla do not have readily ap- A. gondwanicus Szadziewski, 1996: 38. Lebanon. parent homologies with Chironomidae; they Lower Cretaceous are labeled here as sensilla 1, 2, and 4. Sen- A. mcmillani Wirth and Lee, 1958: 337. Australia (Western Australia) sillum 1 has an open apex bearing a tiny peg. A. megaspinus Borkent, 2000a: 381. Lebanon. Sensillum 4 has a porous surface. Lower Cretaceous. The labrum of Austroconops shows a A. sibericus Szadziewski, 1996: 40. Russia. Upper number of similarities to other Ceratopogon- Cretaceous idae. Sensilla 1±4 are clearly present as a group on the labra of most other Ceratopo- The genus Jordanoconops Szadziewski gonidae (Hribar and Mullen, 1991, labeled as was proposed as a monotypic genus to in- ``sensillum styloconicum''). In many Chiron- clude a Lower Cretaceous Jordanian amber omidae the two sensilla SIVA amd SIVB on fossil, Jordanoconops weitschati Szadziews- the anterior portion of the labrum are mor- ki (Szadziewski, 2000). It may actually be a phologically very similar to S2 and S4, and member of Austroconops (see below under 28 AMERICAN MUSEUM NOVITATES NO. 3449

Fig. 20. Structures of third-instar larva of Austroconops annettae;sϭ sensillum. A. Left antenna and dorsal portion of labrum in dorsal view. B. Labrum in anterodorsal view. Scale bars: A ϭ 5 ␮m, B ϭ 2 ␮m. 2004 BORKENT AND CRAIG: BITING MIDGES FROM AUSTRALIA 29

Relationships Between Species of Austro- Midleg tibia without apical spur. Hindleg conops). ®rst tarsomere without thick basal spine or stout setae. Claws simple, each claw apically KEY TO EXTANT SPECIES OF AUSTROCONOPS bi®d. Genitalia: In life, rotated about 90Њ. Borkent (2000a) provided a key to extant Apicolateral process absent. Gonocoxite and extinct males and females of Austrocon- short. Gonostylus mostly straight, tapering to ops. The new species A. annettae may be dis- curved apex, apical spine absent. Parameres tinguished as follows. fused medially. Aedeagus short, setose lobe with ventral plate. Female adult: Descriptive 1. Ratio of male ¯agellomeres 12/13 ϭ 0.62±0.77 statistics in table 2. Head: Ommatidia nar- (®g. 1A); female palpal segment 3 swollen rowly separated dorsomedially. Flagello- in lateral view (®g. 1E); ratio of palpal seg- meres gradually increasing in length from ment 3/ 4 ϩ 5 ϭ 1.36±1.91 for the male, ¯agellomere 2 to 13. Mouthparts moderately 1.43±1.75 for the female; female with only a very slender, setalike tooth on each claw elongate, mandible (®g. 1G) broad, with ®ne (®g. 1I) ...... A. mcmillani teeth, most directed laterally or ventrolater- ± Ratio of male ¯agellomere 12/13 ϭ 0.43 (®g. ally, laciniae with well developed, ®ne re- 1B); female palpal segment 3 relatively slen- trorse teeth. Palpus (®g. 1E) with 4 segments, der in lateral view (®g. 1F); ratio of palpal segment 3 well developed, swollen in lateral segment 3/ 4 ϩ 5 ϭ 1.04 for the male, 1.05± view, with capitate sensilla scattered on sur- 1.32 for the female; female with large tooth face or in shallow pits. Thorax: Scutellum at the base of each claw (®g. 1J) ...... angular in dorsal view. Wing: Costa extend- ...... A. annettae ing to or just beyond apex of R3. Legs: Fem- ora, tibiae slender. Legs lacking armature. Austroconops mcmillani Wirth and Lee Midleg tibia without apical spur. Hindleg Austroconops mcmillani Wirth and Lee, 1958: ®rst tarsomere without thick basal spine or 337. Holotype female, National Park, Perth, stout setae. Foreleg, midleg, hindleg claws Western Australia, Australia, 12-XII-1954 (®g. 1I) slender, evenly curved, each with (ANIC). ®ne inner setalike tooth. Genitalia: Cercus Austroconops mcmillani: Borkent, Wirth and short. Egg: Descriptive statistics in table 3. Dyce, 1987. Male adult. Female adult (in part). First-instar larva: Head capsule length sta- DIAGNOSIS: Male. The only extant Austro- tistics in table 4. Total body length 0.76±1.26 conops with ¯agellomere 12 elongate, 0.6± mm (n ϭ 6). Second-instar larva: Head cap- 0.8 the length of ¯agellomere 13 (®g. 1A). sule length statistics in table 4. Total body Female. The only extant Austroconops with length 1.44±2.13 mm (n ϭ 2). Third-instar simple claws (each with only a very ®ne bas- larva: Head capsule length statistics in table al tooth) (®g. 1I). Egg and larva (all in- 4. Total body length 2.60±3.72 mm (n ϭ 9). stars). Not distinguishable from those of A. Fourth-instar larva: Head capsule length annettae (see generic diagnosis above). statistics in table 4. Total body length 3.78± Pupa. The only known pupa in the genus 4.84 mm (n ϭ 3). Pupa: Total length 2.0 (see generic diagnosis above). mm. General coloration of exuviae, including DESCRIPTION: Male adult: Descriptive sta- respiratory organs, light brown. Integument tistics in table 1. Head: Antenna (®g. 1A) smooth with some ®ne rounded spicules on with well developed plume. Flagellomere 12 dorsal surface of anal segment. Initially, live elongate, with subbasal constriction, 0.62± individuals with reddish hemolymph, after 0.77 length of ¯agellomere 13 (®g. 1A). 24 hours with reddish core of tissue, with Mouthparts moderately long. Palpus (®g. 1C) subcutaneous bluish-green patches in thorax, with 4 segments, segment 3 ovoid in lateral abdomen (further developmental details giv- view, with capitate sensilla scattered on sur- en below). Length of cephalothorax/length of face or in shallow pits. Thorax: Scutellum abdomen posterior to cephalothorax ϭ 0.70. angular in dorsal view. Wing: Costa extend- Dorsal margin of thorax angular in lateral ing to or just beyond apex of R3. Legs: Legs view (®g. 4A). Operculum (®g. 4D) broad lacking armature. Bristles on midleg tibia, dorsally, narrow ventrally, with elongate an- ®rst tarsomere of moderate length (®g. 1M). teromedial seta (®g. 4E) directed anteriorly, 30 AMERICAN MUSEUM NOVITATES NO. 3449

Fig. 21. A. Southwestern margin of fairway 2 at Yanchep National Park where adult Austroconops mcmillani were abundant. B. Western Grey Kangaroo (Macropus fuliginosus) rubbing eye in response to biting females of A. mcmillani at Yanchep National Park. C. Wet area from which adult A. annettae were collected at 5 km SSW of Forest Grove. Arrow points to wet patch; adults swept from tuft of 2004 BORKENT AND CRAIG: BITING MIDGES FROM AUSTRALIA 31 on well developed tubercle; with single pore; posterolaterally directed apicolateral process tubercle situated dorsally on operculum. La- with very apex slightly bent dorsally, bearing bral, mandibular, maxillary, labial sheaths single pore near base; genital sac moderately (®g. 4C) well developed, palpal sheath short, elongate, slightly wider than rest of segment, not extending beyond apex of labial sheath. situated ventrally. Anterodorsal setae (®gs. 4A, 5A), two, both elongate, well developed, lying against dor- DISTRIBUTION AND HABITAT somedial surface of respiratory organ at about its midlength, with pore on dorsal sur- Austroconops mcmillani is known from face of tubercle. Dorsolateral setae (®g. 4F), three sites in southwestern Western Australia three; two elongate, with posterior one slight- (®g. 22B). A fourth locality from ``National ly thicker, on well developed tubercle; third Park, Perth'' cannot be speci®cally located seta short, arising from near base of tubercle. (see Taxonomic Discussion below). Single, small ventromedial seta (®g. 4C). Adult A. mcmillani were collected at ®ve Two ventrolateral setae (®g. 4C), lateral one sites in Yanchep National Park during 2001± longer, thicker. Metathorax (®g. 6A) with two 2002 (®g. 22C: sites A±E). On November setae, one pore, lateral seta bifurcate, elon- 20±21, 2001 adults were extremely abundant gate, medial seta simple, short. Antennal at fairway 2 in the golf course (®g. 22C; site sheath (®g. 5F) apex anterior to apex of por- A). Biting females were very common along tion of midleg lying medial to it. Apex of all the southern margin of the fairway (®g. 21A) legs (®g. 5F) terminating near apex of wing and at times were attacking humans at the sheath. Wing sheath (®g. 5F) rounded api- rate of more than 50 per minute! Further ob- cally, without marginal tubercle. Only apex servations on biting are given below. Swarm- of hindleg (®g. 4A) visible under lateral mar- ing males were collected directly southeast gin of wing sheath. Respiratory organ (®g. of the tee-off area at the southwestern end of 5B, C), on well developed pedicel, length ϭ the fairway, which was the driest area on the 99 ␮m, laterally compressed, with anterior fairway. The southeastern margin of the fair- and slight posterior bulge in lateral view, way had some very wet soil about halfway with 18 pores abutting, arranged in single, down its length, and the northeastern end in- more-or-less longitudinal (somewhat antero- cluded a ditch with open water. The vegeta- laterally to posteromedially) row at apex. tion included the following trees at the west- Dorsomedial (®g. 5A) a minute seta on an- ern drier end: Agonis ¯exuosa (Weeping Pep- terior margin of scutum. Four dorsal setae permint; native to Southwest of W.A., but in- (®g. 5A); setae i, ii, iv well developed, bi- troduced into the Park), Allocasuarina furcate; sensillum vi well developed, simple fraseriana (Common Sheoak), Banksia at- seta. Metathorax (®g. 6A) nearly completely tenuata (Yellow Candle Banksia), Banksia divided medially, with bilobed medial pro- grandis (Bull Banksia), and Banksia menzie- tuberance from scutum protruding nearly to sii (Firewood Banksia). The shrubs and sedg- posterior margin of metathorax. Abdominal es were the same as listed for the eastern segments 2±8 (®g. 4A) with all setae bifur- (wet) end as given below, but were sparse. cate, with a few divided further (®g. 5D, E); Trees at the wet eastern end of the fairway sensilla separate from one another (none on were Banksia littoralis (Swamp Banksia); common tubercle). Following sensilla pre- shrubs were Acacia saligna (Orange Wattle), sent on segment 4 (®g. 5G), all setae on Spyridium globulosum (Basket Bush), Tem- short, well developed tubercles: 2 dasm, i a pletonia retusa (Cockies Tongues), Xanthor- pore, ii a seta; 4 dpm, i, ii, iv setae, iii a pore; rhoea preissii (Balga or Grass Tree) and lasm, 3 lpm, 3 vn all setae; vasm a pore. sedges were Lepidosperma longitudinale Segment 9 (®g. 6B) with well developed, (Pithy Sword-sedge), Lepidosperma striatum

← grass. D. A. mcmillani attacking human ear at Yanchep National Park. E. A. mcmillani attacking human eyelid at Yanchep National Park. F. A. mcmillani attacking human cheek at Yanchep National Park. 32 AMERICAN MUSEUM NOVITATES NO. 3449

TABLE 1 Measurements and Ratios of Male Austroconops Wing measurements in mm.

(no speci®c common name; a sword-sedge), sedge), Schoenoplectus validus (Lake Club- and a herb, Centella asiatica (Indian Pen- rush), and some Typha orientalis and a thick nywort). Samples of mud and detritus from mat of an introduced herbaceous weed Pen- the ditch at the east end of fairway and a few nisetum clandestinum (Kikuyu Grass). Mud wet soil samples from about three-fourths and wet fallen leaves sampled from this site down the fairway failed to produce imma- produced no immatures after ¯oating debris tures. and substrate with sugar or sorting by hand. Austroconops mcmillani adults were also Two females of A. mcmillani were col- observed at the southern margin of Loch lected on November 20 at the northeast cor- McNess (®g. 22C; site B), about 30 meters ner of Loch McNess (®g. 22C; site C). This west of where a small stream enters the lake. area was a very wet, but more open, innun- The adult population was signi®cantly small- dation zone. er than at site A, and on November 18±20 On November 20 from about 10:50 AM to females attacked, at best, at the rate of about 1:15 PM the ®rst author and his wife walked 1±2/minute. One male was swept from the completely around Loch McNess (®g. 22C), surrounding vegetation. This wooded area stopping 14 times at more-or-less even inter- had a narrow innundation zone with very wet vals around the lake for 3 minutes at a time, soil and mud and some small pools that par- breathed deeply (to produce maximum CO2), tially ®lled after a short rain. The predomi- and waited for attacking females. The only nant tree at this speci®c site was Melaleuca spots where females were found were at sites rhaphiophylla (Swamp Paperbark) with B and C (®g. 22C), suggesting that the spe- shrubs of Acacia saligna (Orange Wattle), cies is indeed restricted to some speci®c lo- sedges Carex fascicularis (Tassel Sedge), calities in the park. Females attacked within Lepidosperma effusum (Spreading Sword- 5 minutes at site B when arriving at 2:55 PM

TABLE 2 Measurements and Ratios of Female Austroconops Wing values in mm, spermatheca (largest) in ␮m. 2004 BORKENT AND CRAIG: BITING MIDGES FROM AUSTRALIA 33

TABLE 3 Measurements and Ratios of Eggs of Austroconops Values given in ␮m.

(November 19, 2001) and within seconds EGG LAYING,LARVAL AND PUPAL BEHAVIOR upon arrival at site A at 10 AM (November AND DEVELOPMENT 20, 2001), further indicating the presence of quite localized populations of A. mcmillani. Eggs were laid by female A. mcmillani in All sites shared the presence of very wet a scattered pattern on the surface of wet mud soils, which corresponds well with the be- in a vial or on the sides of the vial just above havior of the reared larvae which actively the mud where moisture had condensed on burrowed through wet mud/detritus. the vial wall. Three eggs of A. mcmillani that were laid on the sides of the vial were at least Additional biting females were collected somewhat dried after they were laid and had by Len Zamudio at fairway 3 on January 3 partially collapsed but when immersed in wa- and 9, 2002 by a small creek which traverses ter and rehydrated, they subsequently hatched. the fairway about halfway down its length The sequence of egg laying, hatching, and (®g. 22C; site D) and at fairway 8 on January larval development is given in table 5. 9 and 31, 2002 (®g. 22C; site E). First-instar larvae burrowed very actively The paratype specimens of A. mcmillani through moderately packed, quite wet mud/ collected at Yanchep N.P. were collected on detritus with a serpentine motion. They used December 23, 1954 by B. McMillan ``at the different means of propulsion depending on opening of a small cave to the right of the the nature of the substrate and whether they track leading north from the golf course'' (A. had located food. When larvae were in water, L. Dyce, personal commun.) and this would but with substrate to crawl upon, they used represent another place in the park where A. their anterior proleg in an anteroposterior mcmillani are known. However, the total motion, dragging the rest of the body along park area has been progressively drying over to produce a slightly jerky anterior move- the past number of years and it is unknown ment. Larvae that were in very loose detritus what impact this might have on the distri- and that wanted to move through the water bution of A. mcmillani in the park since from one clump of detritus to another used 1954. their posterior proleg as an anchor, extending

TABLE 4 Head Capsule Lengths of Larvae of Austroconops Values given in ␮m. 34 AMERICAN MUSEUM NOVITATES NO. 3449

Fig. 22. A. Sketch of a kangaroo indicating the main areas of scratching (eyes, forelegs, backlegs, taut belly of female carrying joey). B. Map of Western Australia showing the distribution of extant species of Austroconops. C. Map of a portion of Yanchep National Park showing the distribution of Austroconops mcmillani within the park. Letters A±E indicate speci®c collection sites referred to in the 2004 BORKENT AND CRAIG: BITING MIDGES FROM AUSTRALIA 35 the head and length of the body through wa- immediately became very active, writhing ter to connect with another piece of detritus. and attempting to ®nd a substrate upon In thicker substrate, at least some movement which to take hold. Upon doing so, they im- was facilitated by serpentine movements of mediately burrowed into the substrate and the anterior half of the body with the abdo- clearly preferred more solid clumps of detri- men being dragged behind and with the pos- tus than ®ne, looser particles to burrow into. terior proleg hooks withdrawn into the anus. Larvae in detritus that were harassed with a In dense substrate (such as thick mud/detritus probe quickly escaped with rapid use of the or agar), the head capsule would additionally anterior prolegs and, in thick substrate, some be extended anteriorly, bent somewhat down undulations of their bodies. to take hold of the substrate and then bent Five ®rst-instar larvae of A. mcmillani farther anteroventrally, helping to bring the were observed feeding on E. coli, and when body forward; these larvae also withdrew doing so they had their posterior proleg ®rm- their anterior proleg into the body in the ly hooked into the substrate and worked over crease between the cervix and remaining acceptable substrates with their mouthparts. prothorax. The proleg was withdrawn much The head capsule moved back and forth, ap- like the inverting of a shirt sleeve, with the pressed to the agar, and the mouthparts terminal hooks inverting ®rst into the interior moved rapidly, with the labrum contracting of the extended leg and being withdrawn to- and apparently moving food into the mouth ward the body and the rest of the proleg fol- cavity. The pharyngeal complex periodically lowing suit. This inversion is almost certain- moved food into the gut. The gut contents ly the result of muscles that insert at the apex appeared white in these individuals (same of the proleg (®g. 4B). Extension of the pro- color as the E. coli). When feeding, the an- leg was the reverse of this sequence. Larvae tennae actively moved independently of one removed from substrate and placed in a drop another. After feeding on E. coli for 3 days, of water on a slide never withdrew the an- all ®ve larvae died. terior proleg as they actively sought a sub- When drops of the fecal infusion, nema- strate to latch on to, and it is clear that this todes, and rotifers were introduced to the lar- structure is important to their locomotion. vae in the mud/detritus substrate, further ob- As larvae were moving through substrate, servations were made of feeding. Larvae their head capsules would move rapidly in a were obviously attracted by the drops of fe- ¯icking motion left and right and dorsoven- cal infusion and were often very concentrat- trally, apparently testing for potential food. ed directly underneath these small pools of The larvae would also periodically stop, at feces. Occasionally, larvae would be seen times anchoring themselves with their pos- lashing at a passing Paramecium or a slower terior proleg and poke their heads rapidly rotifer. A few second-instars were seen to here and there among the detritus, again in successfully attack and eat passing rotifers an apparent search pattern. The larvae rested but missed most that were nearby. The strong regularly and most often used their extended impression of all larval instars was that they posterior proleg hooks as an anchor on the could at best only attack slow prey. substrate. Such resting larvae were generally Second-instar larvae became increasingly in a more-or-less elongate position (with fat and slower and by the third-instar and es- some curves or bends in the body), but oc- pecially by the fourth-instar, larvae moved casionally they would assume a tight U- more cumbersomely, with a slow, serpentine shaped position, with the posterior proleg motion. Serpentine movements of the body hooked into detritus. were obviously the most important basis for When ®rst-instar larvae were removed movement as the larvae worked their way from very wet mud/detritus to water, they through the mud/detritus substrate. To re-

← text. Numbers refer to the fairways at the golf course. D. Map of the type locality of Austroconops annettae at 5 km SSW of Forest Grove, Western Australia. 36 AMERICAN MUSEUM NOVITATES NO. 3449

Fig. 23. Head capsule of Leptoconops gallicus identifying setae (after Clastrier, 1972) (A±C). Female adult left palpus (D±E). A. In dorsal view. B. In lateral view. C. In ventral view. D. Austroconops annettae. E. Leptoconops freeborni. Scale bars: D ϭ 25 ␮m, E ϭ 50 ␮m. verse direction, a larva either simply re- third- and fourth-instar larvae and apparently versed its serpentine movement or, by turn- was exserted only when the larva had dif®- ing its head capsule and anterior portion of cultly ®nding any substrate ahead. Second-, the thorax in a posterior direction, doubled third- and fourth-instar larvae always extend- back along the length of its body The anal ed their anterior proleg in a groping motion hooks were withdrawn into the anus when a when placed in a drop of water on a micro- larva moved and were often (but not always) scope slide and writhed helplessly, very sim- exserted when the larva rested; in some in- ilar to the more ponderous movements of stances the hooks were attached to substrate, Dasyhelea larvae. Larvae which were ha- but in more sparsely distributed substrate rassed with a probe quickly escaped with were simply exserted into the water without rapid serpentine undulations of their bodies being anchored to anything. The anterior through the surrounding substrate. When the proleg also appeared to be rarely used by substrate was shaken and larvae severely dis- 2004 BORKENT AND CRAIG: BITING MIDGES FROM AUSTRALIA 37

Fig. 24. A. Cladogram showing the relationships between fossil and extant species of Austroconops. The Cretaceous fossils Jordanoconops weitschati and A. borkenti share synapomorphy 1 but are not included here (see text for explanation). Bars on the cladogram indicate the age of the species. B. Cladogram showing relationships between extant lineages of Ceratopogonidae. Numbers refer to syna- pomorphies discussed in the text. 38 AMERICAN MUSEUM NOVITATES NO. 3449

Fig. 25. A. Cladogram showing the relationships between early fossil and extant genera of Cera- topogonidae. Numbers refer to synapomorphies discussed in the text. B. Distribution of Austroconops and Culicoides in the fossil record. 2004 BORKENT AND CRAIG: BITING MIDGES FROM AUSTRALIA 39

Fig. 26. A. Larval food and habitats of Culicomorpha. B. Biting periods of adult female Culico- morpha. 40 AMERICAN MUSEUM NOVITATES NO. 3449

TABLE 5 and ¯ooding for 16 minutes (so that the re- Data on Rearing of Austroconops mcmillani spiratory organs were submerged) did not re- (2001±2002) sult in any pupal movement, nor did any very slight jarring of the surrounding substrate. At one point, upon turning on a bright micro- scope light, the pupa brie¯y moved its ab- domen in small circular motions. After 48 hours, the pupa did not look much different from 24 hours earlier. After 72 hours, the in- terior of the pupa became somewhat darker. The blue-green patches were also present in the head (this may have been true earlier but the head could not be seen clearly then). The pupa was very lethargic and moved only slightly when bright light was suddenly shone on it. At 84 hours the developing adult eyes became visible. At 96 hours, the devel- oping adult had darkened further, with the blue-green patches more extensively devel- turbed, fourth-instars often assumed a curled- oped. At 108 hours, the dark adult appeared up position (this was not attempted with ear- to be more-or-less completely developed and lier instars). The darting, probing search pat- its black head and thoracic cuticle could be tern of the head capsule continued in these clearly seen. Shortly before the adult male stages. Larvae sometimes moved their anten- emerged, at 120 hours after pupation, the nae and, when ingesting food, moved their pupa slowly wriggled its way out of its cir- mandibles synchronously. Some second-in- cular burrow and lay on the wet mud surface. star and most third-instar larvae took on a The adult emerged during daylight hours pinkish hue. A few third-instar larvae re- (early afternoon) but failed to completely ex- mained unpigmented and translucent. All pand its wings (they were entrapped by the fourth-instar larvae were pink or reddish. wet mud surface); in all respects it appeared The day before the single fourth-instar lar- normally developed and a hindleg ®rst tar- va pupated, its thorax was strikingly broad, somere length of 176 ␮m (males in nature ϭ the respiratory organs of the pharate pupa 178±214 ␮m, N ϭ 13) indicated that the rear- could be clearly seen, and the eyespots had ing conditions and food provided to the larva moved posteriorly in the larval head capsule. was of suf®cient quality to produce a typical The pupa, which completely shed the larval adult. exuviae, was initially very pale, with red he- molymph, and, at 22±25ЊC, was very slug- BITING ACTIVITY AND BEHAVIOR gish, although capable of very slow circular movements of the abdomen. It was buried in Of those female adults of A. mcmillani bit- the wet substrate at about a 50±60Њ angle ing humans at Yanchep National Park, most from the surface, with the tip of one respi- bit on the face (®g. 21D, F); less than 15% ratory organ just sticking out of the water; of individuals fed elsewhere on the body later, the tips of both respiratory organs were (mostly shoulders, arms, legs). Females on exposed. Throughout its development the the face clearly favored areas without hair pupa had the apex of either or both respira- and, of those attacking the face, were pri- tory organs exposed to air. After 24 hours, marily close to the eyes and directly on the the pupal cuticle appeared nearly transparent ears, with fewer on the cheeks and forehead. (except for the brown respiratory organs), A few fed among hairs right at the hairline, with an inner core of reddish tissue surround- including the scalp, eyebrows, eyelashes, and ed by completely clear ¯uid and a few sub- the ®rst author's beard. We are unable to sug- cutaneous bluish-green patches in the thorax gest the cues used by females to preferen- and abdomen. It was virtually motionless, tially attack the face. When bare arms were 2004 BORKENT AND CRAIG: BITING MIDGES FROM AUSTRALIA 41 held up and appressed against the face, the 27 females studied, total time to complete face was still preferred; however, in such a blood feeding ranged from 1:05 to 4:40 min- position a few females attacked the armpit utes:seconds (mean ϭ 2:42), when ambient (no deodorants used), suggesting that secre- air temperatures ranged from 22 to 29ЊC tions of the skin may be involved as an at- (taken in shade). There was no signi®cant tractant. Slightly elevated heat from these ar- correlation between temperature and biting eas cannot be excluded but this seems un- time, although all the feeding times longer likely, since females were not attracted to than 3 minutes (N ϭ 8) were at temperatures handheld cups of warm or hot water, as are at or below 25ЊC. We found the bite of fe- females of some species of Culicoides and male A. mcmillani to be more painful than Culicidae. It is probable that female A. those of other ceratopogonids that we have mcmillani were attracted by CO2. At the experienced (Leptoconops, Culicoides), per- south end of Loch McNess (®g. 22C; site B) haps because they concentrated on the more when the frequency of biting decreased, the tender facial area. number of incoming females could be in- Females fed only during the day when creased within 3±5 minutes by doing on-the- temperatures were above at least 21ЊC. On spot exercises or by intentionally breathing November 21 females began feeding at fair- deeply and rapidly. way 2 (®g. 22C; site A) at 7:50 AM, when Females ¯ew about the head for a period the ambient temperature was 22ЊC. On No- before landing. Those landing on the hair vember 18, at the south end of Loch McNess nearly always quickly ¯ew off the surface to (®g. 22C; site B), they ceased attacking by resume hovering. After landing on skin, 5:45 PM as the light was fading and the tem- nearly all females began feeding immediately perature was 24ЊC. Females in vials became (very little walking about on the skin sur- lethargic at about 18±19ЊC and at about 16± face). 17ЊC were nearly torpid. It appeared that Kangaroos were closely observed during stronger winds dampened activity, although the ®rst sampling period of October 16±20, females continued to bite in sheltered areas, 2001, before the emergence period of A. such as the leeward sides of trees, a behavior mcmillani. They very rarely scratched or typical of many biting ceratopogonids. High rubbed themselves. When the ®rst author and temperatures (generally above 24ЊC) and in- his wife were experiencing very intense bit- creased humidity appear to greatly increase ing on the golf course in Yanchep N.P. on biting rates (L. Zamudio, personal commun.). November 20±21, it was clear that the kan- Cloudy weather and even a light rain did not garoos were also being severely bothered by stop females from attacking, although a midges (®gs. 21B). Many rubbed their eyes steady drizzle did stop biting activity on the and scratched at their forelegs and the shins afternoon of November 18, 2001. of the hindlegs with their foreleg feet (®g. 22A). Female kangaroos carrying well de- ADULT SEASONALITY veloped joeys (baby kangaroos) also often scratched at the front of their extended Intense collecting of Ceratopogonidae in pouches. The midges attacked the eyes and Yanchep National Park showed that no A. areas with a minimum of hair. The ®rst au- mcmillani adults were present October 16± thor chased kangaroos with an aerial net dur- 20, 2001. November 5 produced moderate ing these observations and collected numbers numbers of females, which slowly increased of A. mcmillani females, but it was uncertain with large numbers biting on November 18. if these specimens were attacking the kan- Biting activity varied, depending on environ- garoos, were attracted to the ®rst author, or mental conditions, until at least January 31, were merely free ¯ying in the area. 2002, indicating a nearly 3-month period of Females assumed a biting posture similar female adult biting activity. Populations in to those of Leptoconops and Culicoides that the 2002±2003 season were substantially less the ®rst author has examined, with the body at Yanchep N.P. (possibly due to a very dry virtually parallel to the skin surface but very season; L. Zamudio, personal commun.) and slightly elevated posteriorly (®g. 21D, F). Of biting female adult A. mcmillani were col- 42 AMERICAN MUSEUM NOVITATES NO. 3449 lected from November 14, 2002 until Janu- cinity of the swarms; the pair separated after ary 1, 2003. Two female A. mcmillani were about 20 seconds while in the net. While still sampled from kangaroo feces on January 21, attached the pair stood on the net mesh fac- 2003 (L. Zamudio, personal commun.), in- ing in opposite directions, showing that the dicating that the emergence period was still male can at least opportunistically rotate his about 3 months long in the 2002±2003 sea- genitalia 180Њ (they are otherwise generally son. held at about 90Њ). Small white sheets (about Records from three years show males have 1 ϫ 1 m) were placed on the ground under been collected Oct. 22±23 and Nov. 1, 1985; the swarms of males but no mating pairs set- Nov. 19±21, 2001; Nov. 20, Nov. 22, Dec. tled on these. The fact that females captured 6, and Dec. 12, 2002. immediately after they had completed feed- ing were able to lay fertile eggs proves that MALE SWARMING mating occurs before feeding.

Male swarms were observed on November OTHER OBSERVATIONS 21, 2001 on fairway 2 of the golf course at Yanchep National Park (®g. 22C; site A) On November 21, 2002 females did not from 6:45 to 9:15 AM. At 6:45 the tempera- attack the ®rst author on fairway 2 in Yan- ture was only 15ЊC (in shade, warmer in sun) chep N.P. before 7:50 AM. From 6:15 until 6: and calm. At 7:50 AM and 22ЊC a single, 45 AM females were sampled by dropping a small (about 30 cm in diameter), more-or- net over one or more fresh (wet) kangaroo less spherical swarm was seen about 1 meter feces and then disturbing the feces with a above the grass and about 6 meters from the stick slipped under the margin of the net that margin of short trees at the edge of the fair- was appressed against the grass. Some fe- way. A light wind began blowing. Several males were sampled by kicking at the feces similarly situated swarms were seen during and then immediately sweeping briskly over the next 40 minutes. The swarms started to the area. In virtually every patch, some fe- increase in size, became more elongate ver- males were collected, with numbers varying tically, and moved to within 3 meters of the from 0 to 8 individuals (58 collected in 30 leeward side of the trees, just south of the minutes). Three attempts at collecting fe- tee-off area. No obvious swarm markers males from dry kangaroo feces failed to pro- were evident, other than being on the lee- duce any specimens. This suggests that the ward side of the trees. By 8:05 AM the bottom females were imbibing nutrients from the of the swarms were 2±3 meters above the wet feces but further observations are re- ground and larger (about 50 ϫ 100 cm) and quired. That reared larvae obtained nutrients withstood some wind, albeit being periodi- from a fecal infusion in the laboratory might cally blown about and then regrouping. suggest the possibility that the females were Swarming activity ceased in this immediate laying eggs on or near the kangaroo feces, area at about 8:30 AM but continued a short but most of the feces were in an area with distance farther east along fairway 2 (where normal soils, and the clearly aquatic larvae it was a bit cooler) and then began in the ®rst of A. mcmillani could almost certainly not area again at 8:37 AM for at least 30 minutes. survive there. One swarm collected at 9:00 AM with a rap- Of 280 preserved male A. mcmillani col- idly swept aerial net consisted of 160 males lected from swarms in Yanchep National and 10 females (the latter may have been Park on November 21, 2001, one specimen captured because females had begun biting had a phoretic female mite intertwined be- by then). By 9:15 AM swarming had ceased tween its legs. The mite belongs to an un- and did not resume for the next 30 minutes, described species of Blattisocius Keegan at which time observations were terminated. (Ascidae) (E. Lindquist, personal commun.). A male and a female of A. mcmillani were This mite genus is virtually cosmopolitan, collected in copula at 8:47 AM during these and 11 of the 12 known species are, or are observations by sweeping from ground level likely to be, predators (the twelfth is an ec- to about 1 meter in height in the nearby vi- toparasite in noctuid moth ears). No obvious 2004 BORKENT AND CRAIG: BITING MIDGES FROM AUSTRALIA 43 internal parasites (e.g., no nematode infesta- mun.). The specimens we examined from Ar- tion) were seen among the specimens ex- madale and 10 miles SE of Darkan are la- amined. beled as paratypes, although they were ex- Female adults clean their antennae with cluded from the type series in the original the foretibial spurs. The wings were cleaned publication. Furthermore, a specimen from by opening the wings very slightly and hook- ``Perth National Park'' (ANIC) was not la- ing one hindleg over the anterior margin of beled as a paratype although it almost cer- the wing and pushing it alongside and under tainly is one. The slide label is written in the abdomen where both hindlegs rubbed W.W. Wirth's handwriting and the specimen their tarsi against each corresponding wing is mounted in his typical fashion; we have surface (so that the left leg rubbed the dorsal added a paratype label to indicate this. surface of the left wing and the right leg The female adults from Yanchep N.P. were rubbed its underside). After wing cleaning, larger than most of the specimens from far- the wings were sometimes left at an angle ther south. The following are the ranges and but shortly after ``clicked'' back to the typi- means of wing lengths for the limited spec- cal overlapping position over the abdomen. imens available: Yanchep N.P., 0.82±0.93 The hindleg tarsi were cleaned by rubbing mm, 0.89 (N ϭ 16); ``Perth N.P.'', 0.80±0.83 them against each other under the body. mm, 0.81 (N ϭ 4); Armadale, 0.71±0.79 (N ϭ 2); Darkan, 0.82 (N ϭ 1). This may indi- TAXONOMIC DISCUSSION cate the possibility of clinal variation, but further specimens and study are required to The adult male and female of A. mcmillani determine this. were described in some detail by Borkent et The pupal dorsal setae are here labeled as al. (1987). There is some confusion about the i, ii, and iv. Based on their position on the exact location of the type locality, recorded thorax, setae i and iv are likely homologous on the labels of the type series as ``National to those named as such in other Ceratopo- Park, Perth, W.A., 21-XII-1954''. There is goninae. However, seta ii could equally be not, and has never been, a national park in homologous to seta iii of the Ceratopogoni- Perth, although the renowned 4-km2 Kings nae. Park may have been a candidate in the mind of the collector. More likely, the labels may SPECIMENS EXAMINED refer to John Forrest National Park, the ®rst national park in Western Australia, located Yanchep National Park, Nov. 18±21, 2001: about 22 km northeast of Perth city center. 442 males, 201 females; Yanchep National An old ``Caltex'' roadmap in the possession Park from females captured Nov. 20, 2001: of the ®rst author, probably from the 1950s 10 eggshells, 2 eggs, 1 eggshell with ®rst- or 1960s, indicates John Forrest N.P. merely instar stuck during emergence, 10 ®rst-instar as ``National Park'', perhaps con®rming that larvae, 2 second-instar larvae, 8 third-instar this is actually the type locality. The original larvae, 3 fourth-instar larvae, 1 pupal exu- collector, B. McMillan, is deceased (A.L. viae and associated male (CNCI); Yanchep Dyce, personal commun.). The holotype is National Park, golf course, Dec. 11, 2001± housed in the ANIC. Jan. 31, 2002: 32 females; Yanchep National Wirth and Lee (1958) recorded the follow- Park, Oct. 22±Nov. 1, 1985: 3 males (ANIC, ing original material, all females: holotype CNCI), 5 females (ANIC, CNCI, WAMP); and 16 paratypes from ``Perth National Yanchep National Park, Dec. 23, 1954: 4 fe- Park'', 9 paratypes from Yanchep National male paratypes (BMNH, BPBM, USNM, Park, 11 non-paratypes from 10 miles SE of WAMP); Yanchep National Park, golf Darkan and two nonparatypes from Arma- course, Nov. 14, 2002±Jan. 1, 2003: 55 dale. We were unable to locate 5 paratypes males, 1833 females; Armadale, Jan. 4, from ``Perth National Park'', 1 paratype from 1936: 2 females (labeled as paratypes) Yanchep National Park, and 10 of the spec- (ANIC, USNM); Darkan, Jan. 29, 1953: 1 imens from 10 miles SE of Darkan, and these female (labeled as paratype) (USNM); Perth appear to be lost (A.L. Dyce, personal com- National Park, Dec. 21, 1954: 3 female para- 44 AMERICAN MUSEUM NOVITATES NO. 3449 types, 1 female not labeled but likely a para- gate, mandible (®g. 1H) narrow, with ®ne type (new paratype label added) (3, USNM; teeth, most directed dorsolaterally, laciniae 1, ANIC). with well developed, ®ne retrorse teeth. Pal- In addition to the above material, more pus (®g. 1F) with 4 segments, segment 3 eggs, eggshells, and specimens of the differ- ovoid, slightly swollen in lateral view, with ent larval instars were studied but were either capitate sensilla scattered on surface or in left to develop or were subsequently lost. shallow pits. Thorax: Scutellum angular in Therefore, the numbers in tables 3 and 4 re- dorsal view. Wing (®g. 1L): Costa extending cording numbers of specimens does not to or just beyond apex of R3. Legs: Femora, match that listed above. tibiae slender. Legs lacking armature. Midleg tibia without apical spur. Hindleg ®rst tar- DERIVATION OF SPECIFIC EPITHET somere without thick basal spine or stout se- The species was named by Wirth and Lee tae. Foreleg, midleg, hindleg claws (®g. 1J) (1958) after the collector of the type series, nearly straight for apical three-fourths, with B. McMillan. stout basal tooth. Genitalia: Indistinguishable from that of A. mcmillani. Egg: Descriptive Austroconops annettae Borkent, statistics as in table 3. First-instar larva: new species Head capsule length statistics in table 4. To- tal body length 0.67±0.77 mm (N ϭ 7). Sec- Austroconops mcmillani: Borkent, Wirth, and ond-instar larva: Head capsule length sta- Dyce, 1987. Female adult (in part). tistics in table 4. Total body length 1.43±1.82 DIAGNOSIS: Male. The only extant Austro- mm (N ϭ 4). Third-instar larva: Head cap- conops with ¯agellomere 12 short, 0.4 the sule length statistics in table 4. Total body length of ¯agellomere 13 (®g. 1B). Female. length 2.05±2.61 mm (N ϭ 8). Fourth-instar The only extant Austroconops with each larva: Head capsule length statistics in table claw with a well developed basal tooth (®g. 4. Total body length uncertain. Pupa: un- 1J). Egg and larva (all instars). Not distin- known. guishable from those of A. mcmillani (see ge- neric diagnosis above). Pupa. Unknown. DISTRIBUTION AND BIONOMICS DESCRIPTION: Male adult: Descriptive sta- tistics in table 1. Head: Antenna with well Austroconops annettae is known from two developed plume. Flagellomere 12 elongate, sites in southwestern Australia (®g. 22B). with subbasal constriction, 0.43 length of ¯a- The type locality (®g. 22D) is on Loop Road, gellomere 13 (®g. 1B). Mouthparts moder- 5 km SSW of Forest Grove, WA. One male ately long. Palpus (®g. 1D) with 4 segments, and four females (one lost after being col- segment 3 slightly ovoid in lateral view lected) were swept from a very small patch (more slender than in A. mcmillani), with of low vegetation immediately beside a very capitate sensilla scattered on surface or in shallow (less than 3 cm deep) small pool less shallow pits. Thorax: Scutellum angular in than a meter in diameter (®g. 21C). The pool dorsal view. Wing (®g. 1K): Costa extending was immediately beside and south of the nar- just beyond apex of R3. Legs: Legs lacking row track of Loop Road and was located in armature. Bristles on midleg tibia, ®rst tar- a dry, shallow streambed (likely with run- somere elongate (®g. 1N). Midleg tibia with- ning water during the winter). Surrounding out apical spur. Hindleg ®rst tarsomere with- vegetation was composed of an open Jarrah± out thick basal spine or stout setae. Claws Marri forest with thick stands of shrubs (Leu- simple, each claw apically bi®d. Genitalia: copogon parvi¯orus was common). In life, rotated about 50Њ. Not distinguishable The single female from Augusta (®g. 22B) from that of A. mcmillani. Female adult: De- was collected with a sweepnet and was either scriptive statistics in table 2. Head: Omma- collected 1.6 km south of Augusta (most tidia narrowly separated dorsomedially. An- likely) or about 1.6 km east of Jewel Cave tenna as for A. mcmillani. Flagellomeres (about 7.5 km NW of Augusta) (D. Colless, gradually increasing in length from ¯agello- personal commun.). mere 2 to 13. Mouthparts moderately elon- Two females found on November 2, 2001 2004 BORKENT AND CRAIG: BITING MIDGES FROM AUSTRALIA 45 at the type locality were collected between 3: TABLE 6 00 and 4:00 PM when the ambient tempera- Data on Rearing of Austroconops annettae ture was 20ЊC (but substantially warmer in (2001±2002) the sun). When the single female retained live in a vial was at 17ЊC, she became very lethargic (nearly torpid), but when the vial was warmed up, she again became very ac- tive. A third female collected on November 4 was lost but was collected at 12:50 PM and at 22ЊC. One male was collected on Novem- ber 13 at 1:15 PM at 33ЊC, and one female was found on the same date and temperature at 3:30 PM. Fervent and repeated sweeping all around this immediate site on each of the above dates failed to produce any further specimens, which suggests that the few adults were indeed concentrated at the small (1977) has shown that the claw shape of fe- pool that appeared to be the only open water males of species of Forcipomyia (Trichohe- (although very small) in a large area. Sam- lea Goetghebuer) are adaptations to cling to ples of mud from the edge and bottom of this the scales of butter¯y wings as females feed small pool failed to produce any immatures. on butter¯y blood (they pierce the wing The distinctive claw of female A. annettae veins to obtain this). If further study shows (®g. 1J), with a pronounced inner basal tooth a relationship between the claw shape of fe- more-or-less situated in the same plane as the males of Austroconops and Leptoconops and rest of the claw, may indicate the type of host vertebrate host type, it will provide the basis upon which the female feeds. Within the for interpreting such variation in the fossil Simuliidae, females of species with a simi- record: Lebanese amber species Austrocon- larly shaped claw are restricted to those ops gladius and A. gondwanicus, 121 million which feed on birds (Crosskey, 1990). With- years old, both have large basal teeth on their in the Ceratopogonidae, the only species of claws similar to those of A. annettae (Bor- vertebrate feeders with variation regarding kent, 2000a). the presence or absence of an inner basal The female allotype collected on Novem- tooth are those in the genus Leptoconops ber 13 laid eggs scattered separately on the (Forcipomyia Meigen (Lasiohelea Kieffer) surface of wet mud in a vial or on the sides and Culicoides all have simple claws with at of the vial just above the mud. The sequence most, a small, very slender spicule). Nearly of egg laying, hatching, and larval develop- all Leptoconops which bite mammals have a ment is given in table 6. Larval behavior was simple claw or a claw with a small basal indistinguishable from that of A. mcmillani spicule. Species with a claw with a pro- (see above). In addition to the feeding ob- nounced inner basal tooth feed either on hu- servations associated with the fecal infusion mans (e.g., L. spinosifrons (Carter) and L. described under A. mcmillani, one ®rst-instar siamensis Carter; Chanthawanich and Del®- larva A. annettae was observed to eat a small nado, 1967) or on birds (L. werneri Wirth live nematode whole (in less than 2 seconds). and Atchley; Wirth and Atchley, 1973), in- A second larva, upon encountering a some- dicating that the shape of the claw may not what moribund large nematode, pierced it at strictly indicate host type in Leptoconops. midlength. This was followed by rapid Other species which have a claw shape vir- movement of the pharyngeal complex and tually identical to A. annettae are L. freebor- ingestion of part of the nematode; it did not ni Wirth, L. melanderi Wirth and Atchley complete feeding on the nematode. In spite and L. patagoniensis Ronderos, but their of these observations, the ®rst- to fourth-in- hosts are unknown. Further research is war- star larvae mostly ignored the large number ranted because we do not have host records of nematodes added to the Petri dishes every for many species of Leptoconops. Lane 2±3 days and, similar to larvae of A. mcmil- 46 AMERICAN MUSEUM NOVITATES NO. 3449

lani, congregated near fresh drops of the fe- DERIVATION OF SPECIFIC EPITHET cal infusion, suggesting that they too primar- ily feed on microorganisms. This species is named after the ®rst au- thor's wife, Annette Borkent. She shared in virtually every aspect of the six-week expe- TAXONOMIC DISCUSSION dition to study Austroconops, and the results of this paper would not have been possible Austroconops annettae is very similar to without her continuous support. A. mcmillani in all its stages (pupa not known) and we were unable to distinguish PHYLOGENETIC CONCLUSIONS differences in male or female genitalia, eggs, or the different larval instars (except for Due to its phylogenetic position in the some size differences in the immatures, family, the genus Austroconops plays a piv- which may have been due to restricted sam- otal role in the interpretation of character ple size or laboratory rearing conditions; ta- states (both morphological and bionomic) bles 3, 4). within the Ceratopogonidae. Because it has The ®rst-instar larvae are described as remained morphologically conservative (un- having nine undivided segments (®g. 2C). like its divergent sister genus Leptoconops), However, in A. mcmillani, early ®rst-instar the character states in this group allow for a larvae also appear to have undivided seg- far better understanding of the diversi®cation ments, whereas older ®rst-instar larvae have of features of other ceratopogonids. Further- divided segments. The same is likely true for more, considering how similar the extant A. annettae. species are to Lower Cretaceous fossils of The female from Augusta was included as Austroconops 121 mya, it seems very likely a specimen of A. mcmillani in the analysis that the biology of the extant species are also by Borkent et al. (1987). similar to those species from so long ago. In short, information on this genus is a window on how at least some Ceratopogonidae were TYPE MATERIAL living at a very early stage of their evolution. Holotype, male adult on microscope slide, There are not many families of extant meta- labeled ``HOLOTYPE Austroconops annet- zoans which include such long-lived genera. tae Borkent, 5 km SSW of Forest Grove, In this section we interpret the phyloge- Loop Road, WA, Australia, 13-XI-2001, A. netic relationships between early lineages of Borkent, CD1998'' (WAMP); allotype, fe- ceratopogonids, followed by an analysis of male adult on microscope slide, labeled as their zoogeography and bionomic diver- for holotype and ``Female which laid eggs, gence. As further characteristics of Cerato- resulting in 1±4 instar larvae'' (WAMP); pogonidae and other Culicomorpha become paratypes 3 females, 7 eggshells, 8 ®rst-instar better known, Austroconops will remain a critically important group to interpret such larvae, 6 second-instar larvae, 9 third-instar features. larvae, 1 fourth-instar larva, 1 terminal por- tion of abdomen of second- or third-instar larva, all reared from eggs laid by female CLADISTIC ANALYSIS allotype; 2 females from type locality but In this section we discuss our hypotheses collected 2-XI-2001 (CNCI); 1 female from of the relationship of species of Austrocon- Augusta, WA, 3-X-1970 (ANIC); immatures ops to each other and the relationship be- (CNCI). tween Austroconops and extant and extinct In addition to the above material, more genera of Ceratopogonidae. Borkent et al. eggs, eggshells, and specimens of the differ- (1987) provided the ®rst cladistic analysis of ent larval instars were studied but were either the relationships between the early lineages left to develop further or were subsequently of Ceratopogonidae. They concluded that, of lost. Therefore, the numbers in tables 3 and extant lineages, Leptoconops was the sister 4 that record specimens do not match that group to all other Ceratopogonidae and that listed above. Austroconops was the sister group of all Cer- 2004 BORKENT AND CRAIG: BITING MIDGES FROM AUSTRALIA 47 atopogonidae other than Leptoconops. These pears to be unique within the Culicomorpha authors, recognizing some con¯icting evi- and is restricted to members of the genus dence, provided an alternative hypothesis, Austroconops (®g. 1K, L). The condition in suggesting that Leptoconops and Austrocon- Leptoconops is impossible to score because ops were sister groups and that, together, all members of this genus lack r-m. There- they formed the sister group of all remaining fore, this derived condition may, in fact, have Ceratopogonidae. As shown below, this al- evolved in the ancestor of the extant Austro- ternative hypothesis is now strongly sup- conops ϩ Leptoconops with subsequent loss ported. in Leptoconops. However, we consider the Since Borkent et al. (1987) provided a fossil genus Minyohelea Borkent, with an cladogram of early lineages, a wealth of fos- oblique r-m, to be the sister group of Lep- sil ceratopogonid material has been described toconops (®g. 25A), and this would indicate from virtually every insect-bearing amber that the derived condition was not present in deposit in the world (see summary in Bor- the ancestor of Leptoconops. kent, 2000a). Some of those fossils, particu- 2. Male tergite 9 with well developed ap- larly those from the Cretaceous, represent icolateral processes (plesiomorphic); ter- other early lineages in the family and their gite 9 with very short or without apicola- cladistic relationships to each other and to teral processes (apomorphic). Apicolateral extant Austroconops and Leptoconops have processes have been repeatedly lost within been analyzed by Borkent (2000a). Members the Ceratopogonidae. Nevertheless, they are of species of both Austroconops and Lepto- present in at least some members of all early conops are present in the oldest amber, from lineages for which males are known. They 121 million years ago (Borkent, 2000a, are present in the earliest lineage of Cerato- 2001). pogonidae, the 121 million-year-old Leban- The discovery of a second extant species oculicoides (Borkent, 2000a). In Leptocon- of Austroconops and, in particular, the im- ops apicolateral processes are present in most matures of the extant species provides the species but are absent or very reduced (leav- means to further test and improve our un- ing only one seta arising from the posterior derstanding of these relationships as present- margin of tergite 9) in the two subgenera ed below. Styloconops Kieffer and Megaconops Wirth and Atchley. Apicolateral processes are also RELATIONSHIPS BETWEEN SPECIES OF present in the fossil genera Archiaustrocon- AUSTROCONOPS ops Szadziewski and Protoculicoides Boesel and are present or absent in Minyohelea In this section we consider the relation- (Borkent, 2000a; Szadziewski, 1996). Within ships between the two extant and six extinct extant genera, they are present in Dasyhelea, species of Austroconops (®g. 24A). As is the absent or reduced to a single seta in Forci- nature of fossils, some character states are pomyia and Atrichopogon Kieffer (a very missing for some species, and this makes our few species with a secondarily derived pair conclusions rather tentative. Nevertheless, of small projections), present in the Culicoi- our hypothesis provides a framework for fur- dini, and present or absent in the remaining ther analysis. The Cretaceous fossils Jordan- Ceratopogoninae. oconops weitschati and A. borkenti are each Further outgroup comparisons were pro- known as a single female and both have the vided by Borkent (1995: char. 3). The pres- derived state of character 1 below. However, ence of apicolateral processes is likely a syn- further features are either not known or are apomorphy of the Ceratopogonidae (see not informative and the two species are not character 9 below in the analysis of generic considered further in the analysis (but see relationships). Their absence (or reduction to comment section below). The following a group of strong setae) in some species of character states were considered. Austroconops is therefore considered to be 1. Wing with r-m transverse or oblique derived, although there is homoplasy in var- (plesiomorphic); r-m more-or-less parallel ious lineages. to R1 and R3 (apomorphic). This feature ap- 3. Wing with costa extending well be- 48 AMERICAN MUSEUM NOVITATES NO. 3449

yond apex of vein R3 (plesiomorphic); cos- 5. Gonostylus with apical spine (ple- ta not extending or only slightly extending siomorphic); gonostylus without apical beyond apex of vein R3 (apomorphic). In spine (apomorphic). This feature is dif®cult , Simuliidae, and early lineages to interpret, mostly because of the dif®culty of Chironomidae (e.g., all Podonominae, of observing the feature in fossil material many Tanypodinae, some Telmatogetoninae; (Borkent, 2000a: char. 8). The feature is pre- Brundin, 1966; Wiederholm, 1989) the costa sent in at least some species of all early lin- extends to and, in early lineages in these eages, other than Lebanoculicoides where the families, past the apex of R4ϩ5. In , condition is uncertain. Within Austroconops, the costa appears to continue along the pos- the condition is known only in A. fossilis and terior margin of the wing. Within the Cera- A. sibiricus where the spine is present. The topogonidae, Lebanoculicoides Szadziewski absence of the spine is therefore considered has a short extension past the apex of R4ϩ5; derived for the two extant species A. mcmil- there is an extension in the earliest lineage lani and A. annettae. of Leptoconops (subgenus Palaeoconops Borkent), but there is none in extant species of this genus, it is present in the fossil genus DISCUSSION Protoculicoides; it is short to long in the fos- This tentative analysis (®g. 24A) indicates sil genus Archiaustroconops; it is absent to that there is a close correspondence between long in the fossil genus Minyohelea;itisab- the age of species of Austroconops and the sent in the monotypic fossil genera Fossilep- pattern of cladogenesis within the genus. The toconops Szadziewski and Alautunmyia Bor- oldest species in Lebanese amber are also the kent; and it is absent in virtually all extant earliest lineages within the genus. The two Ceratopogonidae (Borkent, 1995: char. 46; extant species are sister species; they are also Borkent, 1998). morphologically very similar, with the male It appears, therefore, that a costal exten- and female genitalia and many other features sion is present in at least early lineages of (including all larval features) being presently other families of Chironomoidea and is pre- sent in at least some members of early line- indistinguishable. The congruence between ages of Ceratopogonidae. We therefore con- fossil lineages and patterns of cladogenesis sider the absence of the costal extension in is similar to that present within the entire some Austroconops as derived. family (Borkent, 2000a). 4. Midleg tibia with apical spur (ple- The Jordanian Cretaceous amber fossil siomorphic); midleg tibia without apical Jordanoconops weitschati (Szadziewski, spur (apomorphic). Borkent (2000a) used 2000) may actually be a member of Austro- this feature (as character 13) as evidence for conops, as it shares with Austroconops the the monophyly of Ceratopogonidae other distinctive wing modi®cation described in than Leptoconops and Austroconops. It was character 1 above. The sole feature distin- recognized that the feature was homoplastic guishing it from Austroconops is the loss of in all early fossil lineages (other than the one of the radial cells, and as an autapomor- monotypic Fossileptoconops and Alautun- phy, this loss may have occurred as a sister myia, which either lack the spur or have a group of Austroconops, but equally well may very small one). This suggests that the pres- have occurred within the genus. Jordanocon- ence of the apical spur is likely plesiomorph- ops weitschati belongs between synapomor- ic within the family, with repeated reductions phy 1 and 3 on the cladogram (characters 2, or losses. All Lebanese amber species of 4, and 5 are unknown for this fossil). Rec- Austroconops have the spur, and the state is ognizing Jordanoconops as a valid genus unknown in A. sibiricus and A. borkenti.It leaves Austroconops without any de®ning is de®nitely absent in both extant species of synapomorphy grouping all extant and ex- Austroconops. The loss of a midleg tibial tinct species. For the time being, we are con- spur is therefore considered a synapomorphy sidering it as the sister group of Austrocon- of these two species, although it may actually ops although there is no logical reason to do include both A. sibiricus and A. borkenti. so. 2004 BORKENT AND CRAIG: BITING MIDGES FROM AUSTRALIA 49

RELATIONSHIPS BETWEEN THE EARLY Clastrier and Wirth (as Holoconops kerteszi LINEAGES OF CERATOPOGONIDAE Kieffer) had a L/W ratio of only 3.4, and they drew the egg as short and more-or-less Borkent (2000a) provided the latest cla- oval but reported its shape as ``banana- distic analysis of the relationship between shaped''. They indicated that L. carteri Hoff- Austroconops and other early lineages of man (as L. torrens (Townsend)) had a similar Ceratopogonidae, including those known egg. However, papers on seven other species only as fossils. Our results differ primarily in of Leptoconops reported that these have the our conclusion that among extant taxa, Lep- similar elongate egg as shown here for Aus- toconops and Austroconops are sister groups troconops. We suspect that Smith and Lowe (®g. 24B) and that among genera known as (1948) misidenti®ed the eggs as Leptocon- fossils, Leptoconops, Minyohelea, Austro- ops. conops, Jordanoconops, and Archiaustro- Forcipomyiinae and Dasyhelea have short conops form a monophyletic group (®g. eggs and we consider this a secondarily de- 25A). rived condition within the Ceratopogonidae The relationships between early lineages (apomorphicЉ). of extant Ceratopogonidae are illustrated in 2. Larval head capsule setae o distant ®gure 24B. As is always the case, fossils per- from one another (plesiomorphic); setae o mit study of only a subset of potential char- closely associated, with bases nearly abut- acter states important to the interpretation of ting (apomorphic؅); setae o represented by -their relationships to each other and to the a single seta (apomorphic؆). All Ceratopo extant fauna. We therefore provide a separate gonidae larvae other than Leptoconops have cladogram (®g. 25A) indicating the relation- a pair of setae called ``o'' on the anteroven- ships between all early lineages, including tral area of the head capsule, and in those both extant Austroconops and Leptoconops taxa with two setae, they are close to one and some fossil genera of Ceratopogonidae, another (®gs. 2K, 3G). Larvae of Leptocon- using those characters from below which can ops appear to have only one o seta (®g. 23C). be utilized in their interpretation. Some Forcipomyiinae may also have only As in the previous analysis by Borkent one o seta but others are reported as having (2000a), the monotypic fossil genus Alautun- two; further study is needed. myia is not included here. It is known from The interpretation of this character is two females in New Jersey Cretaceous am- somewhat dif®cult. We have been unable to ber, but some critical character states are ob- accurately homologize the setae on the head scure (Borkent, 1996, 2000b). Its phyloge- capsule of Ceratopogonidae with those of netic placement must await further speci- other Culicomorpha; nevertheless we are mens, including males. con®dent that this is possible, especially The following character states were con- through the study of ®rst-instar larvae and sidered in our analysis. For a given node, innervation patterns. However, other Culi- characters are arranged sequentially accord- comorpha do not appear to have a closely ing to eggs, larvae, pupae, males, and fe- associated pair of setae on the anteroventral males and dorsal to ventral and anterior to area of the head capsule, although the head posterior. capsule setae S9 and S10 of Chironomidae 1. Egg short, oval to somewhat elongate (Saether, 1980) may be homologous to the o (plesiomorphic); egg very elongate (apo- setae illustrated here. In some chironomids morphic؅); egg short (apomorphic؆). Most the two setae S9 and S10 are close to one Ceratopogonidae have very elongate eggs, another. We therefore consider the paired se- with a length/width ratio of more than 5.0 tae o of all Ceratopogonidae, other than those (®g. 2A). Other Culicomorpha have short to of Leptoconops, a synapomorphy for the somewhat elongate eggs, with some Chiron- family. omidae having a maximum L/W ratio of 4.6 The chaetotaxy of the head capsule of Lep- (Nolte, 1993). There are two exceptions toconops has never been homologized with within the Ceratopogonidae. Smith and Lowe those of other Ceratopogonidae, likely be- (1948) reported that the egg of L. arnaudi cause of the rather modi®ed nature of their 50 AMERICAN MUSEUM NOVITATES NO. 3449 head capsules. Based on the illustration of L. and Culicoidea have an apical sieve plate gallicus Clastrier by Clastrier (1972; identi- (with larger holes). Simuliidae and some ®ed as L. kerteszi) we provide a tentative Chironomidae have respiratory organs which identi®cation of the setae (®g. 23A±C). Even function as a physical gill and obtain their if we have not correctly identi®ed seta o in oxygen directly from aerated water (not from Leptoconops, it is appears that there are no the surface). One group of Corethrella Co- setal pairs on the head capsule of any Lep- quillett species in the New World has re- toconops and this is likely derived. The lar- markably ¯attened respiratory organs, each vae of Leptoconops require further study of which has a row of pores along the outer with scanning electron microscopy. edge. The pores are broadly spaced and each 3. Larval pharyngeal complex present has a trachea which gradually merge together and acting as a sieve or absent; not acting near the base of the organ. This condition is as a crushing structure (plesiomorphic); likely independently derived in Corethrella. pharyngeal complex a grinding and sifting 6. Setae on vertex of adult head capsule structure (apomorphic). Borkent et al. scattered or in dorsolateral arrangement (1987: char. 2) discussed this character and (plesiomorphic); in addition to other setae predicted that the then unknown larva of on vertex, a single seta located medially, Austroconops would also have a pharyngeal just dorsal to where the eyes meet or, in complex (as described here: ®gs. 2J, 3F). groups where the eyes are separated dor- However, the pharyngeal complex of Austro- somedially, between these (apomorphic). conops apparently lacks brushes or teeth as This character was discussed by Borkent do some, but not all Leptoconops (Clastrier, (2000a: char. 9) as evidence that Leptocon- 1971; Hribar and Mullen, 1991; Smith and ops was the sister group of all other Cera- Lowe, 1948). Brushes or teeth are present in topogonidae, including Austroconops (i.e., larvae of other ceratopogonid genera but can the derived condition is absent in Leptocon- be dif®cult to detect. ops). This feature requires further analysis 4. Larval anal papillae simple (plesiom- and study since the discovery that Austro- orphic); anal papillae bi®d (apomorphic). conops and Leptoconops are sister groups. Within Ceratopogonidae, the anal papillae Within the Ceratopogonidae, the condition are bi®d in Austroconops (®gs. 3B, 11B), is not known in the fossil genus Lebanocu- nearly all Forcipomyia and Atrichopogon, all licoides. Nearly all Ceratopogonidae other Dasyhelea, and in all Ceratopogoninae than Leptoconops and Fossileptoconops have (poorly known). Leptoconops have simple the apomorphic condition (a few derived spe- papillae (Aussel, 1991; Ishigami, 1959; cies groups have secondarily lost the medial Smith and Lowe, 1948); we consider this as seta; Borkent, 2000a). We are reluctant to in- a reversal to the plesiomorphic condition. terpret the signi®cance of the absence of the Most other Culicomorpha appear to have medial seta in the single known amber spec- simple anal papillae. Simuliidae have anal imen of Fossileptoconops; if true and if the papillae that are apically simple although median seta is present in Lebanoculicoides, many species of have anal papillae the absence in Fossileptoconops may be a which bear a few to many small basal lob- synapomorphy shared with Leptoconops.If ules, clearly a secondarily derived state. Chi- so, it would draw into question the synapo- ronomidae, Thaumaleidae, and Culicoidea morphies proposed here for Leptoconops ϩ larvae have simple anal papillae. These out- Minyohelea (characters 20 and 21). group comparisons support the presence of We have surveyed further extant species bi®d anal papillae as a derived condition. of Leptoconops for this character. It is indeed 5. Pupal respiratory organ with apical true that all are lacking the median seta, porous plate (plesiomorphic); respiratory which is here considered a secondary loss, organ with open pores arranged in a row perhaps related to the adult female habit of (apomorphic). The apomorphic condition is burying itself in sand to rest and lay eggs. nearly unique within the Culicomorpha and As hypothesized here, the presence of the is therefore likely derived. Chironomoidea median vertex seta is a synapomorphy of all other than Ceratopogonidae have a plastron Ceratopogonidae and has been secondarily 2004 BORKENT AND CRAIG: BITING MIDGES FROM AUSTRALIA 51 lost in Leptoconops (and possibly Fossilep- segments 1±8 undivided (plesiomorphic); toconops). For fossil taxa, we have placed abdominal segments 1±8 secondarily di- this feature on the same node as synapomor- vided (apomorphic). The larvae of all in- phies 8±10, even though it is unknown for stars of Austroconops and Leptoconops have Lebanoculicoides (®g. 25A). secondarily divided abdominal segments 1±

-Wing with well developed R4؉5 (ple- 8 (®g. 2D, G). First-instar larvae of Austro .7 siomorphic); R4؉5 thin and faint, very conops do not appear to have the condition poorly de®ned or absent (apomorphic). when they ®rst emerge from the egg (®g. 2C) This was discussed by Borkent (2000a: char. but display the condition upon lengthening. 4), with further modi®cation by Borkent No other Culicomorpha larvae have divided (2001: 8). There are some puzzling patterns segments. Several lineages within the Psy- here. It is clear that there is a strongly and chodomorpha have independently acquired well developed R4ϩ5 in the Lebanese amber the derived condition but these differ in fossil genus Lebanoculicoides, and this pro- structural details, further indicating their in- vides good evidence that this genus forms the dependent acquisition of this feature. The lar- sister group of all other Ceratopogonidae, vae of have secondarily divid- fossil and extant. However, the lesser but still ed abdominal segments but with the anterior clearly evident R4ϩ5 in the Lebanese amber portion shorter than the posterior portion for subgenus Leptoconops (Palaeoconops)is each of these. Larvae of have more dif®cult to interpret. As placed here, secondarily divided ®rst and terminally di- the reduction of R4ϩ5 occurred in the ancestor vided segments with abdominal segments 2± of all other Leptoconops, and its complete 6 divided into three divisions. Some Psy- loss occurred independently in at least three chodinae also have secondarily divided ab- other lineages (®g. 25A) and possibly more, dominal segments with associated sclerotized depending on the relationships of unresolved plates to indicate their position. lineages. Although some Leptoconops larvae have 8. Hindtibia with single transverse row only abdominal segments 1±8 subdivided ;tibial comb of some au- (Aussel, 1991; Laurence and Mathias, 1972 ؍) of apical spines thors) present or absent (plesiomorphic); Smith and Lowe, 1948), other species appear hindtibia with two transverse rows of to have further divisions of thoracic and/or spines (apomorphic). This feature was dis- abdominal segments (Clastrier; 1971, 1972). cussed by Borkent (2000a: char. 1). All Leptoconops larvae have a well devel- 9. Male adult tergite 9 without apicola- oped cervix as a subdivision of the protho- teral process or, if present, lacking setae rax, as do most (all?) Ceratopogoninae. (plesiomorphic); pair of apicolateral pro- 12. Fourth-instar larvae without hemo- cesses present and each bearing at least globin in hemolymph (plesiomorphic); one seta (apomorphic). This character is with hemoglobin (apomorphic). Fourth-in- discussed above as character 2 in the analysis star larvae of Austroconops have a red he- of the relationships between species of Aus- molymph which is assumed here to be due troconops. to the presence of hemoglobin. This feature 10. Genital fork (sternite 9) of female appears to be unique with the Ceratopogon- genitalia well developed, with vaginal apo- idae. However, at least some larvae of Lep- deme extending along dorsal wall of com- toconops have orange hemolymph, which is mon oviduct (plesiomorphic); vaginal apo- here considered to be homologous to that of deme very reduced or absent (apomorph- Austroconops: L. americanus Carter (as L. ic). This character was discussed by Borkent kerteszi, Rees et al., 1971), L. bequaerti et al. (1987: char. 1). The feature was not (Kieffer) (as Leptoconops sp., Painter, 1927), clearly visible in any Lebanese amber fossils. L. arnaudi (as Holoconops kerteszi in Smith It will be particularly important to discover and Lowe, 1948), and L. gallicus (as L. ker- if it is present in the Cretaceous lineage Le- teszi, Clastrier, 1972). Leptoconops borealis banoculicoides (as the earliest lineage of Gutsevich larvae are pale yellow (Krivosh- Ceratopogonidae, ®g. 25A). eina, 1962). Other Leptoconops larvae have 11. Larval instars 1±4 with abdominal been described as being entirely white: L. al- 52 AMERICAN MUSEUM NOVITATES NO. 3449 biventris de Meijere (Aussel, 1991), L. bezzii ber. Nearly all other Ceratopogonidae have (NoeÁ) (Dzhafarov, 1962), L. carteri (as L. ®ve segments but there have been a number torrens, Smith and Lowe, 1948), L. irritans of reductions in unrelated taxa (listed by Bor- (NoeÁ) (Clastrier, 1971), and L. spinosifrons kent et al., 1987: 598); also in Diaphanob- (Laurence and Mathias, 1972). It is possible ezzia Ingram and Mac®e, in some Forcipo- that at least some of these reports of white myia, and in some Atrichopogon). larvae were due to examination of preserved Within fossil lineages there are reversals material (hemoglobin being no longer red to the plesiomorphic condition in one of the when preserved), but at least some of these six fossil species of Austroconops (A. gond- reports must be accurate (e.g., authors who wanicus) and in four out of the six known examined live larvae). We interpret these species of Archiaustroconops (A. szadziews- pale larvae as having secondarily lost the kii Borkent, A. hamus Borkent, A. bocapar- pigmentation. The reddish hemolymph of vus Borkent, A. alavensis Szadziewski and fourth-instar larvae Austroconops and those Arillo). of Leptoconops with orange hemolymph 15. Male and female adult with palpal generally appear during the third-instar, al- segments entire (plesiomorphic); apex of though a few second-instar larvae of A. palpal segment 3 and base of segments 4 mcmillani were pale pink. ؉ 5 membranous (apomorphic). The adults Culicomorpha larvae with red or orange of both species of Austroconops and nearly hemolymph are otherwise known only in all species of Leptoconops have a membra- some Chironomidae (some Tanypodinae, nous connection at the apex of segment 4 and many Chironominae), where this feature is base of segment 4 ϩ 5 (®g. 23D, E). This almost certainly independently derived. condition is not known elsewhere in the Cul- 13. Male antennal ¯agellomere 13 with- icomorpha and is therefore a synapomorphy out subbasal constriction (plesiomorphic); of these two genera. The derived condition, ¯agellomere 13 with subbasal constriction although present in nearly all species of Lep- (so ¯agellomere has a basal bulb) (®g. 1A, toconops, appears to be absent in a few spe- B) (apomorphic). Borkent et al. (1987: 597) cies (e.g., L. asilomar Clastrier and Wirth, L. discussed this feature and stated that it was stygius Skuse), which we consider to be re- unique within the Chironomoidea. However, versals. within the Culicoidea, a similar condition ap- The feature unfortunately is not clear in pears in the and some Culici- the fossil material the ®rst author has ex- dae. amined and therefore cannot be presently in- Among fossil taxa, this feature is present terpreted for those taxa. in males of species of Archiaustroconops. 16. Larval mandible positioned laterally Males of Jordanoconops and Fossileptocon- on head capsule (plesiomorphic); mandi- ops are unknown. Minyohelea is considered ble positioned more medially (apomorph- the sister group of Leptoconops on the basis ic). The mandibles of Leptoconops are of other character states (see characters 20 unique in the family in lying side by side and 21 below) and is therefore considered to (®g. 23C). All others are more separated me- have lost the derived condition in the sce- dially. Other Culicomorpha larvae have mod- nario presented here. erately to widely separated mandibles with 14. Male and female adult with ®ve pal- most operating in opposition or at an oblique pal segments (plesiomorphic); with four angle. palpal segments (apomorphic). Extant 17. Larval mandible with short to mod- adult Austroconops and Leptoconops have erately elongate apodeme, simply attached only four palpal segments (®gs. 1C±F, 23D, to base of mandible (plesiomorphic); man- E), with segments 4 and 5 being fused (as dibular apodeme very elongate, anteriorly evidenced by the landmark capitate sensilla broken into separate parts (apomorphic). on segment 3). Culicomorpha other than Cer- Larvae of Leptoconops have an unusual and atopogonidae have 5 palpal segments with elongate apodeme extending from near the only a few representatives of Chironomidae, base of the mandible posteriorly to inside the and many Culicidae having a reduced num- prothorax, and this feature is unique in the 2004 BORKENT AND CRAIG: BITING MIDGES FROM AUSTRALIA 53

Culicomorpha. The apodeme has been re- those of Austroconops (the sister group of ferred to as a ventrolateral rod by previous Leptoconops) are pointed (®g. 6B), as are authors. The anterior portion of the mandib- those of all other Ceratopogonidae, we con- ular apodeme of Leptoconops appears to be sider the condition in Leptoconops as de- separated into two or more different portions rived. Outgroup comparisons support this in- intimately associated with the base of the terpretation. Chironomidae have a wide array mandible, identi®ed by Smith and Lowe of apicolateral processes from pointed to (1948) as the mandibular lever, by Clastrier membranous; the pointed condition, howev- (1971, 1972) as ``plaques de l'arc geÂnien'' er, is present in many, including early line- (sclerites of the genial arch), and as the sub- ages (Brundin, 1966; Wiederholm, 1986). genal sclerite by Laurence and Mathias Simuliidae, , , and (1972). These small apodemes may actually most but not all Thaumaleidae pupae have be part(s) of the mandible itself. This com- pointed apicolateral processes (called termi- plex of apodemes at the base of the mandible nal spines in Simuliidae). Chaoboridae and is also unique within the Culicomorpha. The Culicidae have articulated, membranous pu- homology of the ``ventrolateral rod'' with the pal paddles in place of the apicolateral pro- mandibular apodeme of other Culicomorpha cesses. The rounded, lobelike apicolateral is con®rmed by its intimate connection with processes of Leptoconops appear to be the base of the mandible (via the small an- unique in the Culicomorpha. terior separate portions) and by the presence 20. Wing with long radial cells (plesiom- of large adductor muscles attached to the orphic); radial cells very short (apo- ``ventrolateral rod'' which serve to adduct morphic). This character was discussed by the mandible (Laurence and Mathias, 1972). Borkent (2000a: char. 12) but was presented The larva of Austroconops also has a rel- as a derived condition independently evolved atively long mandibular apodeme (®gs. 2I, in Leptoconops and Minyohelea. We here 3E). Larvae of other Ceratopogonidae have consider it to be a synapomorphy of these either a very short apodeme or lack them al- two genera. This character may merely be together. Chironomidae are variable, but the another expression of synapomorphy 21. Podonominae we examined had short apo- However, there are other Ceratopogonidae demes. We have not yet made further out- (derived lineages) in which reduction of the group comparisons but these may show a radial cells does not produce a stigma (e.g., more elongate apodeme to be a synapomor- Baeohelea Wirth and Blanton, Baeodasymyia phy of Leptoconops and Austroconops, with Clastrier and Raccurt, and Nannohelea Gro- the apodeme becoming longer and more gan and Wirth). modi®ed in Leptoconops larvae. 21. Female wing with R1 and R3 joining 18. Larvae with four anal papillae (ple- costa without thickening (plesiomorphic); siomorphic); with two anal papillae (apo- female wing with R1 and R3 joining costa morphic). Leptoconops larvae are unique as a strong, thickened stigma (apomorph- within the Culicomorpha in having only two ic). Borkent (2000a: char. 6) considered this anal papillae (Aussel, 1991; Ishigami, 1959). feature to have evolved independently in Larvae of all other Ceratopogonidae and Leptoconops and Minyohelea. Because of the nearly all other Culicomorpha have four of discovery that Austroconops is the extant sis- these structures. The larvae of Simuliidae ter group of Leptoconops, we considered have three and those of Chironomidae have them to be homologous here. The condition either two, four, or six, but nearly all have is better developed in some Minyohelea spe- four (almost certainly the plesiomorphic con- cies than in others (Borkent, 2000a: ®gs. 19f, dition in that family). Thaumaleidae and all 20a). Culicoidea have four anal papillae. 22. Wing with r-m (plesiomorphic); 19. Pupal apicolateral processes pointed wing lacking r-m (apomorphic). The wing (plesiomorphic); apicolateral processes of Leptoconops, in lacking a r-m, appears to rounded (apomorphic). Leptoconops is the be unique within at least the Culicomorpha. only ceratopogonid genus whose pupae have 23. Female antenna with 13 ¯agello- rounded apicolateral processes. Because meres (plesiomorphic); female antenna 54 AMERICAN MUSEUM NOVITATES NO. 3449 with 10±12 ¯agellomeres (apomorphic). to have evolved independently; their anten- This character was discussed by Borkent nae are each situated on a cuticular tubercle (2001). The feature may be used only as ev- and either do not appear to have any sensilla idence for the monophyly of extant Lepto- or have very short sensilla restricted to the conops subgenera. Within this genus, the ex- very apex of the antenna (Jeu and Rong, tinct Lower Cretaceous subgenus Palaeocon- 1980). Within Forcipomyia only the distinc- ops has 13 ¯agellomeres. tive larvae of the subgenus Phytohelea 24. Posteromedial margin of female Remm have short antennae, which seems al- sternite 8 with or without semicircular most certainly to be a secondary reduction. concavity, without semicircular row of Saunders (1957; as F.(Trichohelea)) stated stout setae (plesiomorphic); with semicir- that the antennae are reduced to'' windows'' cular concavity bearing four or more stout which bear minute sensilla. Larvae of species setae on its margin (apomorphic). The apo- of Kieffer have somewhat morphic condition is unique within at least elongated blades (not nearly as long as those the Culicomorpha. All female Leptoconops of Austroconops), but this likely represents examined have the feature (Clastrier and convergence. Boorman, 1987; Herzi and Sabatini, 1983; Outgroup comparisons indicate that other Wirth and Atchley, 1973). Culicomorpha have elongate antennae and 25. Female cercus short, rounded in lat- many Chironomidae have an elongate blade, eral view (plesiomorphic); cercus elongate, perhaps indicating that the condition in Aus- laterally compressed (apomorphic). This troconops is plesiomorphic and that the short feature was discussed by Borkent (1995: antenna and blade in Leptoconops is inde- char. 6) but as a synapomorphy for only pendently derived from those of Dasyhelea some Leptoconops. Subsequent discoveries and Ceratopogoninae. of Lebanese amber Leptoconops (Borkent, 27. Larval head capsule with seta q (ple- 2001), which are the sister group of all extant siomorphic); without seta q (apomorphic). Leptoconops, showed that elongate cerci are As discussed above, the lack of seta q in Aus- basal to the genus and that the short cerci of troconops (®gs. 2H, 3C) may be a misinter- the females in the subgenera Styloconops and pretation. It may be that seta t is missing and Brachyconops Wirth and Atchley are rever- that q is actually more anteriorly placed in sals to the plesiomorphic condition. Austroconops. Regardless, either seta q or t The condition is unique within the Cera- is absent in Austroconops and, because both topogonidae, and within the Culicomorpha of these setae are present in other Cerato- occurs only in members of the Telmatoge- pogonidae, this is likely a synapomorphy. toninae (Saether, 1977), which are clearly The homology of these setae with those in specialized (marine) Chironomidae. The other Culicomorpha is not yet understood. elongate cerci of some Leptoconops species 28. Larval head capsule with posterior are likely used to oviposit in sand. Females seta of setae p simple, tapering (plesiom- of at least some Leptoconops in the subgenus orphic); with posterior seta of setae p sub- Styloconops, which have short cerci, lay their apically expanded (®gs. 7A, B, 12A, 18) eggs directly on the sand surface (Aussel, (apomorphic). Within the Ceratopogonidae 1993). the apomorphic condition is otherwise 26. Larval antenna short, with short known only in some Forcipomyia species blade (plesiomorphic); antenna long, with where it almost certainly secondarily de- elongate blade (apomorphic). Within the rived; the shape of the single seta p (see char- Ceratopogonidae, larvae of nearly all species acter 45 below) is very diverse in different have short antenna and a short, squat, some- species of Forcipomyia. Chironomidae, Sim- what bulbous blade. Larvae of Austroconops uliidae, and Culicoidea do not have apically have an elongate antenna and a strikingly expanded dorsal head setae and only some elongate antennal blade (®gs. 2H, 3C, G, 9A, Thaumaleidae have plumose setae. B, 14A). The larvae of most species of For- 29. Apex of pupal palpus extending to cipomyia and all Atrichopogon have very level of or posterior to apex of labium (ple- elongate antennae, but we considered these siomorphic); apex of palpus anterior to 2004 BORKENT AND CRAIG: BITING MIDGES FROM AUSTRALIA 55 apex of labium (apomorphic). This feature ately long setae (plesiomorphic); with very is nearly unique within the Culicomorpha elongate setae (apomorphic). The derived (®g. 4C). In Chironomidae the palpus is di- condition, present only in Fossileptoconops, rected laterally and therefore cannot be appears to be unique within the Culicomor- scored here, although they are long enough pha. that if they were directed posteriorly, they 34. Male adult antennal pedicel with too would extend beyond the apex of labium. large basal foramen (plesiomorphic); ped- 30. Pupa with simple setae (plesiom- icel with moderately sized or narrow basal orphic); dorsal setae 1, 2, 4 and all setae foramen (apomorphic). This feature, dis- on abdominal segments 2±8 bifurcate cussed by Borkent (1995: char. 17), could be (some divided into 4) (apomorphic). The seen clearly in only one specimen of the fos- derived condition is nearly unique within the sil genus Protoculicoides (Borkent, 2000a) Ceratopogonidae (®gs. 4A, 5A, G). Only two and it had the plesiomorphic condition. Un- species of Kieffer (Borkent and fortunately, this character is impossible to Craig, 2001) and Pellucidomyia leei Wirth observe in most fossils. have bifurcate setae and these are almost cer- 35. Pupa with hindleg sheath not visible tainly secondarily derived. The bifurcate se- or barely visible along lateral margin of tae of the pupa of Austroconops mcmillani wing sheath (plesiomorphic); hindleg are bifurcate at their bases while in these oth- sheath clearly visible (apomorphic). Within er taxa, the setae are bifurcate at midlength the Ceratopogonidae, all pupae other than or beyond. The pupa of A. annettae is un- those of Austroconops (®g. 4A) and Lepto- known and so this synapomorphy may ac- conops have the hindleg sheath clearly visi- tually be an autapomorphy of A. mcmillani. ble along the lateral margin of the wing Within the Culicomorpha, only Corethrel- sheath. Blanton and Wirth (1979: ®g. 35B, lidae, Chaoboridae, and Culicidae have some C) provided a good illustration of a Culicoi- plumose seta. A few genera of Chironomidae des pupa showing this feature as the sclerite have pupae with some plumose setae but between the ``wing pad'' and ``abdomen 2''. most have simple setae. Simuliidae and In Austroconops and Leptoconops (Clastrier, Thaumaleidae all have simple setae. 1972: ®g. 4B; Ishigami, 1959: ®g. 8; Laur- 31. Pupa with anterodorsal setae short, ence and Mathias, 1972: ®g. 9) there is only or if more elongate, not abutting against membrane present between the wing sheath respiratory organ (plesiomorphic); anter- and second abdominal segment, and the hin- odorsal setae elongate and abutting dleg is completely under the wing sheath (ex- against respiratory organ (apomorphic). cept for its apex medially). The apomorphic condition appears to be Other Culicomorpha exhibit some varia- unique in A. mcmillani (®gs. 4A, 5A). The tion. Within the Chironomidae, virtually all position of the two strong anterodorsal setae have the wing sheath not visible. However, abutting against the respiratory organs clear- in the Podonominae genus Boreochlus Ed- ly functions to support the respiratory organs wards the hindleg sheath is apparently pre- and keep them from bending either medially sent (Brundin, 1966: 307); other Podonomi- or posteriorly. Why this might be necessary nae have the sheath hidden. In other Culi- in nature is not known. comorpha, the sheath is either not visible or 32. Wing veins with r-m transverse or is present (e.g., Simuliidae). We consider the oblique (plesiomorphic); r-m more-or-less situation in Boreochlus and Simuliidae to be parallel to R1 and R3 (apomorphic). The convergent with that present in the Forcipo- feature appears to be unique within the Cul- myiinae ϩ Dasyheleinae ϩ Ceratopogoninae. icomorpha and is restricted to members of 36. Adult thoracic anapleural suture the genus Austroconops and the extinct Cre- well developed, extending to anterior mar- taceous genus Jordanoconops. See character gin of anepisternal cleft (plesiomorphic); 1 and the comment section of the analysis anapleural suture short, extending to pos- between species of Austroconops for further terior margin of anepisternal cleft (apo- discussion. morphic). This character was discussed by 33. Female cercus with short to moder- Borkent (2000a: char. 14). The feature is 56 AMERICAN MUSEUM NOVITATES NO. 3449 probably an excellent indicator of relation- but this is certainly due to homoplasy. Other ship; no instances of homoplasy are known. Culicomorpha do not have a large pharyn- All Lower Cretaceous ceratopogonids have geal complex in the center of their head cap- the plesiomorphic condition. sules. 37. Trochanter of fore- and midleg with 42. Larval salivary duct opening near only slender, simple setae (plesiomorphic); margin of hypostoma (plesiomorphic); sal- trochanter of fore- and midleg each with ivary duct opening near center of head pair of thick, contiguous setae (apomorph- capsule (apomorphic). Lieven (1998) ic). This character was discussed by Borkent showed that, within the Chironomoidea, this (2000a: char. 15). feature is restricted to Forcipomyiinae and 38. Adult midleg tibia with spur (ple- Dasyhelea. It would be valuable to study ad- siomorphic); midleg tibia lacking spur ditional taxa within and outside of the family (apomorphic).This feature was discussed by to con®rm this. Furthermore, this character Borkent (2000a: char. 13). No member of may be another expression of character 41, this lineage has a midleg tibial spur. The de- in which the pharyngeal complex is situated rived condition is susceptible to homoplasy in the middle of the head capsule. in the outgroup, with numbers of losses (see 43. Male antenna with setae on ¯agel- character 4 in the analysis between Austro- lomere 1 of similar length to those on sub- conops species above). sequent ¯agellomeres (other than those 39. Larval head capsule prognathous few terminal ¯agellomeres which have (plesiomorphic); head capsule at least par- shorter setae) (plesiomorphic); setae on tially hypognathous (apomorphic). The lar- ¯agellomere 1 much shorter than those on vae of Dasyhelea are partially hypognathous more distal ¯agellomeres (apomorphic). and those of nearly all Forcipomyiinae are This feature was discussed by Borkent completely hypognathous. Only members of (2000a: 403). Although there is some ho- the subgenus F.(Phytohelea) are progna- moplasy in higher lineages within the Cera- thous and those of Forcipomyia (Warmkea topogoninae, this feature appears to be a Saunders) are only partially hypognathous; good indicator of relationship. because other Forcipomyia and all Atricho- 44. Larvae with short, slender setae pogon are hypognathous, these are probably (plesiomorphic); with prominent, strong reversals. It seems likely that the completely setae (apomorphic). Forcipomyiinae larvae hypognathous condition is actually a syna- are unique within the family in having prom- pomorphy of the Forcipomyiinae, but a cla- inent, strong setae on the head, thorax, and distic analysis of the Forcipomyiinae is re- abdomen. Many taxa have these setae on quired to con®rm this. Outgroup compari- raised tubercles. Within the Culicomorpha, sons indicate that all other Culicomorpha are strong setae are also present in Thaumalei- prognathous, except for the larvae of Thau- dae, which is likely an independently derived maleidae, which are also hypognathous, and feature. this must certainly be due to homoplasy. This character was discussed by Borkent 40. Larval mandibles articulating with et al. (1987: char. 9) as ``setae arising from head capsule (plesiomorphic); mandibular cuticular projections''. Some Forcipomyia do articulation reduced (apomorphic). Lieven not have cuticular projections, although these (1998) showed that the mandibles of larvae are almost certainly reversals. of Forcipomyiinae and Dasyhelea are 45. Larval head capsule with two setae uniquely disarticulated. p (plesiomorphic); with one seta p (apo- 41. Larval head capsule with pharyn- morphic). Larvae of all Ceratopogonidae, geal complex moderately developed (ple- other than those of Atrichopogon and nearly siomorphic); pharyngeal complex massive all Forcipomyia, have two setae p. Only two (apomorphic). This feature (Lieven, 1998) is species in the highly modi®ed subgenus F. nearly unique within the Ceratopogonidae. (Phytohelea) have two setae p and this must Larvae of species of Culicoides (Monoculi- be considered a reversal (Chan and LeRoux, coides Khalaf) also have large pharyngeal 1971; F. nicopina Chan and LeRoux, F. complexes (Murphree and Mullen, 1991), grandis Chan and LeRoux). Further outgroup 2004 BORKENT AND CRAIG: BITING MIDGES FROM AUSTRALIA 57 comparisons to other Culicomorpha are not acter was discussed by Borkent (1995: char. yet possible because we are uncertain of ho- 25). mologies. 51. Male and female adult scape lacking 46. Larval antenna close to anterior ventral apodeme (plesiomorphic); scape margin of head capsule (plesiomorphic); with ventral apodeme (apomorphic). This antenna on basal half of head capsule character was discussed by Borkent et al. (apomorphic). Forcipomyiinae are nearly (1987: char. 11). unique in the Culicomorpha in having pos- 52. Male adult ¯agellomeres lacking teriorly displaced antennae. Within the genus longitudinal striations (plesiomorphic); only members of the modi®ed F.(Phytohe- ¯agellomeres with longitudinal striations lea) have anteriorly placed antennae, almost (apomorphic). This character was discussed certainly a case of reversal. Within the Cul- by Borkent et al. (1987: char. 10). icomorpha, only Thaumaleidae, which super- 53. Male and female with foretibial spur ®cially resemble Forcipomyiinae, have pos- (plesiomorphic); without foretibial spur teriorly positioned antennae. (apomorphic). This character was discussed This character may be an expression of the by Borkent and Craig (1999, as character 3). head capsule having become hypognathous 54. Larvae swim with a slow thrashing (see character 39 above) and perhaps is not motion (plesiomorphic); larvae swim with an independent indicator of relationship. very rapid serpentine motion (apomorph- 47. Adult terminal ¯agellomere tapering ic). Mullen and Hribar (1988) pointed out or rounded apically (plesiomorphic); ter- that the rapid swimming motion of most Cer- minal ¯agellomere with basally constrict- atopogoninae is unique among Diptera. ed nipple at apex (apomorphic). This fea- Some lineages within the subfamily have ture is unique within the Culicomorpha. secondarily lost this motion. However, it is However, some species of Atrichopogon do worth noting that some larger larvae have a lower swimming beat rate than do their ear- not have the derived condition, which we in- lier instars (Linley, 1986), and comparing terpret as a reversal to the plesiomorphic earlier and later instars may indicate that state. only later instars have lost the serpentine 48. Male adult tergite 9 with pair of ap- swimming ability. Regardless, it is clear that icolateral processes present and each bear- this behavior has been lost by some lineages ing at least one seta (plesiomorphic); with- (e.g., Culicoides (Avaritia Fox)). out apicolateral processes or each repre- Other Ceratopogonidae move on sub- sented merely by a single seta (apomorph- strates either by crawling (Forcipomyiinae, ic). The plesiomorphic feature is considered some Dasyhelea) or with a serpentine move- a synapomorphy of the Ceratopogonidae ment (Leptoconops, Austroconops, some (character 9 above). Many Forcipomyia have Dasyhelea). a pair of setae on the posterior margin of 55. Larva with posterior proleg (ple- tergite 9 but others lack even these. Forci- siomorphic); without posterior proleg pomyia generally do have posterior projec- (apomorphic). Larvae of Leptoconops and tions, sometimes drawn as arising from the Ceratopogoninae are the only members of posterior margin of tergite 9, but in fact these the family without a posterior proleg, and we are the cerci. consider these as independent losses. Poste- 49. Adult thoracic paratergite without rior prolegs with hooks are present in all oth- setae (plesiomorphic); paratergite with 1± er Chironomoidea. In Corethrellidae and 6 setae (apomorphic). This character was Chaoboridae they are reduced so that only discussed by Borkent (1995: char. 22). In- apical hooks are present (Cook, 1956). stances of homoplasy within the family 56. Pupae with at most one seta and one (Kaczorowska, 2000: 104) are certainly in- pore on anterior portion of abdominal ter- dependently derived (restricted to Palpomyi- gite 4 (plesiomorphic); with two setae ini and Stenoxenini). (apomorphic). Nearly all Ceratopogoninae 50. Egg ovoid to elongate (plesiomorph- pupae have two anterior setae on abdominal ic); egg c-shaped (apomorphic). This char- tergite 4 (called dasm for Ceratopogonidae). 58 AMERICAN MUSEUM NOVITATES NO. 3449

The only exception are species of Parabezzia cipital ridge and a horizontal tentorium and Malloch, where only one seta is present; pu- this was interpreted as evidence of the sister pae of this genus are otherwise quite modi- group relationship between these two line- ®ed. Other Ceratopogonidae have either one ages. This character state is in con¯ict with seta, one pore, or one seta and one pore (®g. the conclusions here and requires further 5G). Other Culicomorpha generally have study, especially of the state in further Chi- only one or no seta. Only within Chiron- ronomidae and other Culicomorpha. omidae are there some taxa with more setae Our new analysis also results in signi®cant scattered on the tergite (Wiederholm, 1986), changes in the interpretation of fossil genera, but at least within the Podonominae, there recognizing for the ®rst time the following appears to be only one anterior seta (Brun- as a monophyletic group: the extant Austro- din, 1966; personal obs.). conops and Leptoconops and the fossil gen- 57. Sternite 9 of female terminalia form- era Minyohelea, Jordanoconops, Archiaus- ing a continuous band ventrally (plesiom- troconops, and Fossileptoconops (®g. 25A). orphic); sternite 9 discontinuous medially, Of these, Leptoconops and Minyohelea are forming two halves (apomorphic). This now considered as sister groups. There is one feature was discussed by Borkent (1995: previously published character state which char. 26). suggested different phylogenetic relation- ships. Borkent (2000a) and Szadziewski DISCUSSION (1996) considered a hindleg tarsal ratio/fore- leg tarsal ratio of Ն1.40 as derived and evi- When Wirth and Lee (1958) ®rst described dence for the monophyly of the Austrocon- Austroconops and puzzled over its classi®- opinae, at that time including Austroconops, cation, they stated that ``Final disposition of Archiaustroconops, and Minyohelea. Consid- the genus Austroconops would best await the ered to be a nearly unique feature within the discovery of the male or, perhaps better still, family, Borkent (2000a) noted there was the immature stages.'' Our discovery and in- some homoplasy within Forcipomyia. Since terpretation of the eggs, larvae, and pupae then, further instances of homoplasy with amply con®rm their prediction! Among ex- values as high as 1.8 have become evident tant taxa, we demonstrate here that Austro- within some extant and some fossil Lepto- conops and Leptoconops are sister groups. Of conops (Szadziewski, in press). We now these two genera, Leptoconops is overall a therefore no longer recognize any synapo- highly derived group, while Austroconops is morphies for Austroconopinae. generally plesiomorphic. Other relationships shown in ®gure 24B In the ®rst cladistic analysis of the Cera- are those that have been hypothesized earlier topogonidae, there were two character states (Borkent, 1995), but we have added substan- proposed by Borkent et al. (1987) to unite tial support through the addition of further Austroconops with all Ceratopogonidae other characters, especially from the immatures. In than Leptoconops. We now consider the ®rst particular, the monophyly of Forcipomyiinae character state, the presence of a medial adult and Dasyhelea is con®rmed. This is espe- head capsule seta in Austroconops and all cially important because two characters giv- Ceratopogonidae other than Leptoconops,to en by Borkent (1995) have since been shown have been secondarily lost in Leptoconops to be suspect (Borkent, 2000a: 403). In ad- (see character 6 above). The second character dition, the Ceratopogoninae have further sup- state is problematic. Borkent et al. (1987) port as a monophyletic group. A recent mo- pointed out that the postoccipital ridge was lecular study of the mitochondrial cyto- relatively poorly sclerotized and that the pos- chrome oxidase subunit 2 of a variety of Cer- terior portion of the tentorium was directed atopogonidae by Beckenbach and Borkent in a more posterodorsal angle in adult Lep- (2003) supported all the relationships shown toconops and that this was also the condition in ®gure 24B. present in Chironomidae (hence plesiom- In our analysis here we have excluded the orphic). Austroconops and remaining Cera- monotypic genus Washingtonhelea Wirth topogonidae have a more sclerotized postoc- and Grogan, which may form the sister group 2004 BORKENT AND CRAIG: BITING MIDGES FROM AUSTRALIA 59 either of the Forcipomyiinae ϩ Dasyhelea, of mere of the hindleg was a synapomorphy of the Ceratopogoninae, of Forcipomyiinae ϩ all Ceratopogonidae other than those known Dasyhelea ϩ Ceratopogoninae, or potentially as Lower Cretaceous fossils. That was in er- of a subgroup of the Ceratopogoninae (Bor- ror and the feature, as discussed here, is re- kent, 1995). Female Washingtonhelea from- stricted to those Ceratopogoninae which meri Wirth and Grogan have broadly serrate form the sister group of the Culicoidini. mandibles, a character state restricted to a The male adults of A. mcmillani and A. subgroup within the Ceratopogoninae, sug- annettae have permanently erect antennal gesting that the genus may be a member of plumes. All other male Ceratopogonidae the Ceratopogoninae. It will be fascinating to have erect plumes only when they are in a examine the features of the immatures of this sexually aroused state during swarming pe- genus once they are discovered. riods. Indeed, this is the basis for the name It is striking that we have been unable to of the family: Ceratopogon Meigen means identify a synapomorphy for Forcipomyia. ``bearded horn'', referring to the decumbent We suspect that the genus will be found to male antennal plume. In other Culicomorpha, be paraphyletic in relation to Atrichopogon. most families with males possessing a plu- There is a wealth of character states in both mose antenna (Chironomidae, Corethrellidae, of these genera, and a cladistic analysis is Chaoboridae) have the plumes permanently long overdue. Such an analysis would further erect. Male Anopheles Meigen also have de- test or re®ne some character states used in cumbent plumes but other mosquitoes have our analysis here (characters 2, 26, 39, 45± permanently erect plumes. These outgroup 48). comparisons suggest that the decumbent state Because of their phylogenetic conclusions, has either evolved twice in Ceratopogonidae Borkent et al. (1987) proposed that Austro- (in Leptoconops and most Ceratopogonidae) conops be placed in the new subfamily Aus- or the plesiomorphic condition has been re- troconopinae. The discovery here that the ex- acquired by Austroconops. Nijhout and Shef- tant Leptoconops and Austroconops and the ®eld (1979) described the mechanism by extinct Minyohelea, Jordanoconops, Ar- which male Anopheles erect their plume, and chiaustroconops, and Fossileptoconops form it would be a test of homology to see if the a monophyletic group negates the necessity same is present in Ceratopogonidae. of recognizing a separate subfamily Austro- In our analysis we reported only 12 syn- conopinae, and we here place all these gen- apomorphies for the genus Leptoconops.In era in the subfamily Leptoconopinae (new reality, it would be possible to report many synonym). more by interpreting each detail of the larval There are several features of ceratopogon- and pupal chaetotaxy and more details of the ids discussed in earlier publications which larval mouthparts (maxillae, buccal cavity, require some comment. Borkent (1995, etc.) and other modi®cations. It is clear that 2000a) argued that the presence of an ante- Leptoconops is highly derived in many of its rior larval proleg must be plesiomorphic features. within the Ceratopogonidae because it is pre- Our proposal that Leptoconops and Min- sent in all other Chironomoidea and ®rst-in- yohelea form a monophyletic group suggests star Corethrellidae (Borkent and McKeever, that the extinct Minyohelea had elongate 1990). As predicted by Borkent (1995: 103± eggs (character 1), larvae with secondarily 104), Austroconops larvae have an anterior divided abdominal segments (character 11), proleg. Therefore, within the Ceratopogoni- and hemolymph with hemoglobin (character dae the anterior proleg has been indepen- 12), as well as those features shared by all dently lost by Leptoconops,byDasyhelea Ceratopogonidae (characters 2±4). and by the Ceratopogoninae (®rst-instar lar- vae of Culicoides species have the feature ZOOGEOGRAPHY but it is lost in the second-instar; Jobling, 1953). Although the two extant species of Aus- Borkent (2000a: char. 17) indicated that troconops are presently restricted to the the row of palisade setae on the ®rst tarso- southwestern corner of Western Australia, 60 AMERICAN MUSEUM NOVITATES NO. 3449 the genus was much more broadly distributed tures in each life stage. Indeed, this was a in the past, with fossils recorded from Leb- primary motivation to discover the imma- anese, Spanish, French, Siberian, and Bur- tures; the adults were earlier recognized as mese ambers. There are two interesting dis- conservative, and it was expected that the tributional features of these fossils. First, immatures would be the same. True to ex- Austroconops fossils are known only from pectations, the features of the immatures Old World ambers. Second, all Austroconops were easily homologized with those of other fossils are known only from the Cretaceous, Ceratopogonidae, especially those of Dasy- dating from 85±121 mya (®g. 25B). By 44 helea and Ceratopogoninae (Forcipomyiinae mya, Austroconops had been eliminated from larvae and pupae are rather divergent). We at least northern Europe, as suggested by its suggest that this is why Austroconops now absence in Baltic amber (with 1103 speci- has such a restricted distributionÐthe Cera- mens and 101 ceratopogonid species record- topogoninae, and especially species of Culi- ed; Szadziewski, 1988, 1998). Ceratopogon- coides, have retained many of the basic fea- idae from other Old World Tertiary deposits tures of Austroconops and have replaced either remain unidenti®ed or include too few Austroconops throughout most of its historic specimens to be interpreted here. Austrocon- range. There are two pieces of evidence sup- ops, therefore, was diverse in Old World Cre- porting this assertion. First, there is a general taceous ambers and sometime since its last pattern of mutual exclusion in the fossil re- record 85 mya it became restricted to West- cord (®g. 25B). Austroconops, as noted ern Australia. above, is restricted to amber 85±121 million What might be the reasons for the present years old. The earliest records of the Cera- day relict distribution of Austroconops?We topogoninae, including the genus Culicoides, suggest that the answer lies in an understand- are in New Jersey amber (88±93.5 mya), ing of the nature of the adaptations of the which does not contain Austroconops. Sibe- early lineages of Ceratopogonidae. It is strik- rian amber from Yantardakh (83±88 mya) in- ing that the genus Leptoconops, as the sister cludes the last fossil record of Austroconops group of Austroconops, is known from every and the ®rst Old World record of Culicoides, signi®cant amber deposit (Old World and and we suggest that this provides a snapshot New), except for the relatively recent depos- of the transition from the one genus to the its on the Dominican Republic (which oth- other. As noted earlier, Baltic amber at 44 erwise have a reduced ceratopogonid fauna; mya includes an abundant and diverse group Borkent, 2000a), and is found today world- of Ceratopogoninae and no Austroconops. wide. Leptoconops is an example of an an- Second, mutual exclusion between Austro- cient lineage that has survived and thrived conops and Culicoides is also suggested in by developing a peculiar life cycle with an the present day local distribution of Austro- accompanying wide array of autapomor- conops. Although Culicoides is otherwise di- phies. Species are restricted to areas of beach verse in Western Australia, we did not en- (or alkaline soils) where the larvae burrow counter any biting Culicoides at Yanchep Na- through the sand, and forage on microorgan- tional Park during the entire adult season of isms; in many places they are the dominant A. mcmillani (sampled for 2 years). Neither macroorganism in this specialized habitat. did we encounter biting Culicoides at the Adult females of most (perhaps all) species type locality of A. annettae, and this further rest by burrowing into sand, either at night suggests that, to this day, the two lineages for unfed females or both day and night after exclude one another. blood-feeding. The larval, pupal, and adult Considering that the oldest New World stages all have unusual features and have di- amber is from New Jersey (88±93.5 mya) verged so much that it is dif®cult to homol- (®g. 25B), it is quite possible that Austro- ogize many of the larval and pupal features conops was present in the New World before with those of other ceratopogonids or Culi- they were, as hypothesized here, eliminated comorpha. On the other hand, Austroconops through competition by the evolution of the is in many ways the quintessential ``living Ceratopogoninae (particularly Culicoides). If fossil'', preserving many plesiomorphic fea- ceratopogonid-bearing amber older than 95 2004 BORKENT AND CRAIG: BITING MIDGES FROM AUSTRALIA 61 mya is discovered in North America, we sug- the general pattern within the family (®g. gest that it will include Austroconops. In ex- 26A). The larvae of Leptoconops, Forcipo- amining maps of land distributions from the myiinae, Dasyhelea, and many early lineages Cretaceous (Barron, 1987), it is clear that the within the Ceratopogoninae (most Culicoi- Cretaceous islands of Europe that preserved des, many ) feed on micro- amber were close to coastal New Jersey organisms, fungi, and algae. Some Dasyhe- (with its 88±93.5 mya amber) and there were lea also feed on dying or decayed inverte- also connections between eastern Russia and brates and one species is a leaf-miner (per- western North America at that time. All am- sonal obs.). Many members of the ber is preserved in coastal environments and Culicoidini and Ceratopogonini appear to in- it would be hard to suggest what factor could clude both microorganisms and small prey keep early to mid-Cretaceous Old World organisms in their larval diet. Ceratopogonid Austroconops out of the New World. Finally, species that are completely predaceous as lar- it will be particularly fascinating to see if vae appear to be restricted to the lineage Austroconops shows up in the 53 million- composed of the Heteromyiini ϩ Sphaerom- year-old Eocene amber of Le Quesnoy, iini ϩ ϩ Stenoxenini. It is clear France (Nel el al., 1999) or the 57±65 mil- therefore that feeding on microorganisms is lion-year-old Sahkalin, Russia amber (Szad- plesiotypic within the family. Within the Chi- ziewski, 1990). Our hypothesis here suggests ronomidae all early lineages feed on micro- that Austroconops should not be present in organisms and predation is restricted to all any Tertiary amber deposits in the Holarctic Tanypodinae, some Chironominae, and a few region because Culicoides was well estab- Orthocladinae. The larvae of all Thaumalei- lished throughout the region by that time. dae, Simuliidae, and Dixidae are also brows- If the above pattern is correctly interpret- ers on microscopic organisms and small or- ed, we suggest that there may be a good basis ganic particles. Within the Culicoidea it ap- for the mutual exclusion between Austrocon- pears that predation evolved at least four ops and Culicoides. It was striking to us that times: in Corethrellidae, Chaoboridae, and, larvae of Austroconops moved relatively within the Culicidae, at least some Culicinae slowly, both in seeking out food and in re- (e.g., some Psorophora Robineau-Desvoidy) sponse to disturbance, when compared to and all Toxorhynchitinae. It thus seems likely those of most species of Culicoides. In ad- that feeding on microorganisms is plesiotypic dition, larvae of Austroconops could not for the entire Culicomorpha. swim, again in contrast to the distinctive, re- Although we were unable to discover the markably fast swimming motion of Culicoi- larval habitat of Austroconops species in na- des (and most Ceratopogoninae). Such swim- ture, it was clear from laboratory rearings ming abilities allow Culicoides larvae a rapid that the larvae require wet, loose detritus/ escape response, as well as dispersal to other mud in which to move about and feed. If the nearby habitat. We speculate that Culicoides mud compacted at all, the larvae experienced larvae may have a distinct selective advan- dif®culty in moving about. When placed in tage to those of Austroconops but, of course, free-standing water the larvae became fran- this needs to be tested. tic, looking for attachment on any piece of detritus; upon contact with a substrate they BIONOMIC DIVERGENCE immediately began burrowing. This strongly suggests that the larval habitat in nature must The larval feeding habits of Ceratopogon- be a permanently wet substrate that would idae are quite diverse, ranging from grazing not be disturbed by either current or wave on microorganisms to being obligate preda- action. Such a restricted aquatic habitat tors, eating either whole prey or attacking would be congruent with the pattern other- and invading the larval bodies of much larger wise present within the family. Ceratopogon- Chironomidae (Hribar and Mullen, 1991; id larvae occur in a wide variety of semi- Mullen and Hribar, 1988). The discovery that aquatic and aquatic habitats, ranging from Austroconops larvae could be reared on mi- decaying vegetation and wet mosses to rivers croorganisms present in a fecal infusion ®ts and lake bottoms. However, nearly all mem- 62 AMERICAN MUSEUM NOVITATES NO. 3449 bers of the early lineages (®g. 26A) are pre- Borkent (2000a) pointed out that both Lep- sent in small or restricted habitats, such as toconops and Austroconops species are xer- rock pools, epiphytes, and wet soils at the ically adapted (only a very few species of margins of lotic and lentic habitats. The im- Leptoconops occur in more mesic areas). matures of Leptoconops are generally found Considering that the two genera are now in wet or damp sand or sandy soil near fresh- known to be sister groups, it is likely that water or marine beaches, although a few spe- this adaptation was present in their common cies are known from seepage areas in desert ancestor as well as in the extinct genus Min- oases, the wet margins of salt ¯ats, the mar- yohelea (®g. 25A). gin of vernal ponds in desert areas, in halo- There is a pattern of dispersal capabilities morphic, calcareous soil, or in cracked clay among the early lineages of Ceratopogoni- soils. Forcipomyiinae and Dasyhelea are ei- dae. Borkent (1991) showed that the cerato- ther in small aquatic habitats or live in or on pogonid fauna of volcanic oceanic islands wet vegetation (many feeding on algae). All worldwide is virtually always restricted to of the early lineages within the Ceratopogon- species of four genera: Forcipomyia, Atri- inae are either in small aquatic or very wet chopogon, Dasyhelea and Culicoides. These habitats (e.g., wet soil and pools) or on the genera include many species which live in margins of rivers and lakes. Only some spe- ephemeral habitats, such as small bodies of cies of the lineage Heteromyiini ϩ Sphae- water and wet, decaying vegetation, and it romiini ϩ Palpomyiini ϩ Stenoxenini truly was concluded that these temporary habitats occur in rivers and lakes. Small aquatic hab- selected for taxa which were good dispersers. itats and/or wet areas appear to the plesi- All four genera are distributed worldwide. otypic habitat for the Ceratopogonidae. Out- They also are early lineages within the Cer- group comparisons suggest that this feature atopogonidae (®g. 24B), suggesting that fre- is actually plesiotypic for the entire Culico- quent dispersal was an early characteristic of morpha. The pattern within the Chironomi- the ancestor of the Forcipomyiinae ϩ Dasy- dae is somewhat uncertain for two reasons. heleinae ϩ Ceratopogoninae (Culicoides is First, the most recent phylogenetic interpre- an early lineage within the Ceratopogoni- tations of the family (Saether, 2000) included nae). The lineage Leptoconops ϩ Austrocon- character states that have not been polarized ops shows a different pattern. The absence of using outgroup comparisons. Second, the Leptoconops species on nearly all islands re- Chironomidae are markedly diverse in their ¯ects the permanency of its primary habitat larval habitats. We agree with Cranston et al. on marine and freshwater beaches, even (1987) that the earliest Chironomidae were though it has an extensive fossil record ex- present in small lotic and lentic habitats and tending to 121 mya. The restriction of Aus- that presence of spiracles in these taxa is ple- troconops to Western Australia likewise sug- siomorphic. Further outgroup comparisons gests that it is a weak disperser and that its are more clear. The earliest lineage of Sim- larval habitat, once found in nature, will be uliidae, the genus Malloch, is a predictable, stable environment. The ances- restricted to small seeps, and Thaumaleidae tor of Leptoconops and Austroconops there- are restricted to wet rock faces. Within the fore was likely in a stable habitat as well and Culicomorpha, nearly all are restricted to was a poor disperser. small aquatic habitats. Only some species in One of the striking features of female adult the derived Chaoboridae genus Chaoborus Austroconops is that they are diurnal biters. are found in lakes; presence in lakes is likely To interpret this feature phylogenetically, ®g- a plesiotypic feature of Chaoborus because a ure 26B shows the distribution of the times number of the larval features are clearly re- when female adult Culicomorpha bite. Vir- lated to predation pressures by ®sh. Other tually every lineage includes species which genera of Chaoboridae, representing earlier do not feed at all. Within the Ceratopogoni- lineages in that family, are restricted to small dae, female adult Leptoconops, Austrocon- pools and ponds. This pattern shows that ops, and Forcipomyia (Lasiohelea) (the only presence in small aquatic habitats is a plesi- Forcipomyia that feeds on vertebrates) all otypic feature of the entire Culicomorpha. feed during the day. Other Forcipomyia and 2004 BORKENT AND CRAIG: BITING MIDGES FROM AUSTRALIA 63

Atrichopogon feed on larger , and lim- thermic hosts (Crosskey, 1990). Perhaps the ited records indicate that they too attack their original hosts of biting Culicomorpha were hosts during the day. Female Dasyhelea do ectotherms and detection of their hosts was not suck blood, and Culicoides females may only possible when hosts were active during attack at any time (species' behaviors vary the day (producing more CO2 and visually within the genus). Other biting Ceratopogon- apparent). Similar to Culicidae (Bock and inae feed on insects of approximately the Cardew, 1996), female adult Culicoides, in- same size (up to two to three times their own cluding those that feed nocturnally, use a length), with most entering male swarms of combination of olfactory, visual, and thermal other , males of their own spe- cues to detect their hosts (Kline and Lemire, cies, or, for a few species, attacking Ephem- 1995). It would be interesting to know what eroptera. They may be diurnal or crepuscular. else attracts Leptoconops and Austroconops This pattern within the family strongly sug- females to their hosts, other than their known gests that diurnal feeding is probably plesi- attraction to CO2 (Brenner et al., 1984) and otypic within the family and that the crepus- especially if they are attracted by heat. Our cular and nocturnal feeding periods of some interpretation here would predict that they Culicoides and other Ceratopogoninae are a are not attracted by thermal cues. derived behavior. Both species of Austroconops almost cer- Outgroup comparisons suggest that diurnal tainly overwinter in the larval stage (tables feeding is plesiotypic to at least the Chiron- 5, 6) and this is by far the most prevalent omoidea. The feeding period of the only bit- mode of Ceratopogonidae in temperate areas. ing Chironomidae (Archaeochlus Brundin, Only a few species of Culicoides appear to Austrochlus Cranston) are unknown, but overwinter in the egg stage (Jobling, 1953; Simuliidae bite only during the day. The Cul- Parker, 1950). Overwintering in the larval icoidea are either nocturnal feeders (Coreth- stage is therefore plesiotypic within the Cer- rellidae bite calling frogs), feed at variable atopogonidae. times (Culicidae), or do not feed at all (Dix- idae, Chaoboridae). SUGGESTIONS FOR FUTURE What might be the signi®cance of diurnal RESEARCH feeding in Simuliidae and early lineages of As always in science, more information Ceratopogonidae? The oldest fossil Culico- means more questions. This study indicates morpha are members of the Chironomidae several areas where further investigation and are recorded from the Upper would be fruitful. (Krzeminski and Jarzembowski, 1999). As the Chironomidae are the sister-group of the ● Locate the larvae and pupae of Austroconops Ceratopogonidae, the latter must be consid- species in their natural habitat. This would ered equally as old, and the earlier diver- give a better understanding of the adaptations gence of the Simuliidae shows that the black- of these species and enhance the likelihood of conserving these ``living fossils''. ¯ies must have been present in the Upper ● Describe the pupa of A. annettae to test the Triassic as well. This pattern suggests that purported pupal synapomorphies detailed the diurnal biting habit shared by early lin- here. eages of Ceratopogonidae and Simuliidae ● Investigate the presence of female adults of originated during or, more likely, earlier than A. mcmillani on wet kangaroo feces and de- the Upper Triassic. The Middle and Upper termine the signi®cance of that behavior. Triassic was the time during which the di- ● Determine whether Austroconops transmits nosaurs and mammals, respectively, ®rst any diseases between its hosts. The host of arose. The diurnal feeding habit may, there- A. annettae is unknown. fore, re¯ect an early stage of bloodfeeding, ● Determine further homologies with other Culicomorpha. Comparison of the ®rst-instar in which Ceratopogonidae and Simuliidae larvae of Culicomorpha would be particular depended primarily on visual cues and de- valuable because these seem to generally be tection of heightened CO2 production by more conservative. Within the Ceratopogon- their daytime hosts. Apparently extant Sim- idae, an SEM study of the larva and pupa of uliidae do not use heat to detect their endo- a species of Leptoconops would allow ho- 64 AMERICAN MUSEUM NOVITATES NO. 3449

mologies of this divergent group to be better golf course, suggested that female adults understood. were concentrated in the morning hours on ● Study of Forcipomyiinae cladistically. fresh kangaroo feces, had important infor- ● A comparative morphological study of the mation concerning feeding by the females, larvae and pupae of Ceratopogonidae is basic and graciously collected further samples, al- to good keys to the genera and would pro- vide further phylogenetically important char- lowing for an estimation of adult periodicity. acters (Borkent, in prep.). This study would have been much poorer without his enthusiasm and help. Dr. Alan L. Dyce (ANIC) provided much ACKNOWLEDGMENTS appreciated and detailed advice regarding Annette Borkent, the precious wife of the this project. Previously described specimens ®rst author, not only supported this trip with of Austroconops were loaned by Dr. David ®nances but bravely withstood the onslaught Yeates (ANIC), Dr. Terry Houston (WAMP), of biting midges, smiled while being bitten Nancy Adams (USNM), Dr. Neal Evenhuis and photographed, and shared numerous as- and Keith Arakaki (BPBM), Dr. Jeff Cum- pects of the exciting expedition to Australia ming (CNCI), and Nigel Wyatt (BMNH) and that led to the information reported here. The we thank these curators for their efforts. The ®rst author wonders what he would do with- Canadian Food Inspection Agency of the out her! Government of Canada kindly gave permis- The expedition to Australia was supported sion to import live species of Austroconops. ®nancially by four other sources, and the ®rst George Braybrook (University of Alberta) author is grateful for their substantial help: provided highly skilled and much appreciat- the Dipterological Fund (Canada), the Smith- ed assistance with our SEM study of larval sonian Institution and S.W. Williston Diptera specimens. The E. coli and C. elegans col- Research Fund, CanaColl Foundation (Can- onies were sent by Dr. Tetsunari Fukushige ada), and the Schlinger Foundation. (University of Calgary) and we very much The Australian National Park service was appreciate his assistance with this project. Judy Acreman (University of Toronto, Cul- incredibly helpful and supportive. In partic- ture Collection of Algae and Cyanobacteria) ular, John Wheeler (Ranger in Charge, Yan- kindly sent cultures of blue-green and green chep N.P.) and Paul Tholen (Yanchep N.P.) algae as potential food for larvae of Austro- provided logistical support and a wealth of conops. The single mite found on an adult information without which this study could Austroconops mcmillani was identi®ed by not have taken place. Additionally, park Dr. Evert Lindquist (CNCI, Ottawa). rangers in other national parks provided ex- A gracious neighbor of the ®rst author, cellent support and good advice concerning Tom Marshall, repeatedly cared for live Aus- collecting sites. The Department of Conser- troconops larvae over the period of nearly a vation and Land Management (CALM) pro- year when the ®rst author needed to leave vided the ®rst author with collecting permits home (ϭ laboratory) for periods of time; in Western Australia. without his patient and much appreciated Dr. Terry Houston at the Western Australia help the larvae could not have been reared Museum in Perth readily lent much appre- to the fourth-instar and A. mcmillani right ciated ®eld equipment during the stay in through to the adult stage. Tom and Mara Australia and gave important advice concern- Moen, friends of the ®rst author living down ing collecting sites and we thank him for this the road, cheerfully supplied the goat feces effort. David T. Pike (Wild¯ower Society of which were instrumental in providing food Western Australia) identi®ed the ¯ora at the for the larvae. collecting sites in Yanchep National Park. We express our sincere thanks to C. Steven We express our deep appreciation and Murphree and William L. Grogan, Jr. for thanks to Len Zamudio who works at the their careful reviews of this paper. golf course in Yanchep National Park and who took a special interest in the ®rst au- REFERENCES thor's research there. He ®rst pointed out Aussel, J.-P. 1991. Isolement et description de la concentrations of female A. mcmillani at the larve de Leptoconops (Styloconops) albiventris 2004 BORKENT AND CRAIG: BITING MIDGES FROM AUSTRALIA 65

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a This paper meets the requirements of ANSI/NISO Z39.48-1992 (Permanence of Paper).