A new cryptic species of Brazilozoros Kukalova-Peck & Peck, 1993 from French Guiana (Zoraptera, Spiralizoridae) Author Kočárek, Petr Department of Biology and Ecology, University of Ostrava, Chittussiho 10, CZ- 710 00 Ostrava (Czech Republic) petr. kocarek @ osu. cz (corresponding author) ivona. horka @ osu. cz kocarek@osu.cz Author Horká, Ivona Department of Biology and Ecology, University of Ostrava, Chittussiho 10, CZ- 710 00 Ostrava (Czech Republic) petr. kocarek @ osu. cz (corresponding author) ivona. horka @ osu. cz kocarek@osu.cz text Zoosystema 2023 2023-03-21 45 6 163 175 journal article 10.5252/zoosystema2023v45a6 1638-9387 7774863 urn:lsid:zoobank.org:pub:B1DD23BB-DA32-4DA3-BB1C-7B6AB05871F0 Brazilozoros kukalovae n. sp. ( Figs 1-4 ) urn:lsid:zoobank.org:act: F896840D-F6EE-4BA2-8293-80A47942990B TYPE LOCALITY . — French Guiana : Montagne des Singes. MATERIAL EXAMINED. — Holotype . French Guiana • ♂ ; French Guiana , Kourou env., Montagne des Singes ; 5°4’17.11”N , 52°41’50.26”W ; 40 m a.s.l. ; 7.VI.2022 ; P. Kočárek , M. Jankásek , I. H.Tuf leg.; NMPC . Paratypes . French Guiana • 1 ♀ ; same data as for holotype; NMPC • 1 ♂ ; French Guiana , Montsinéry env., Sentier du Bagne des Annamites track; 4°49’37.23”N , 52°30’58.24”W ; 25 m a.s.l. ; 11.VI.2022 ; P. Kočárek , M. Jankásek , I. H. Tuf leg.; NMPC • 1 ♂ , 1 ♀ ; French Guiana , Cacao env., Molokoï track; 4°33’39.70”N , 52°27’44.52”W ; 40 m a.s.l. ; 10.VI.2022 ; P. Kočárek , M. Jankásek , I. H. Tuf leg.; MNHN . DIAGNOSIS. — Brazilozoros kukalovae n. sp. ( Figs 1-4 ) is morphologically similar to B. weidneri ( New, 1978 ), but males of the two species can be distinguished by the specific arrangement of sensilla basiconica on 10th tergite and by the shape of the male genitalia. The anterior group of sensilla basiconica on the 10th abdominal tergite are composed of four sensilla in B. kukalovae n. sp. , and the anterior sensilla are apparently shorter than those of the posterior group ( Fig. 3D ). The anterior group of sensilla basiconica are composed of only two sensilla in B. weidneri , anterior sensilla are longer than those of the posterior group; lateral sensilla basiconica of anterior group are divided basally ( Fig. 3E ). Differences in the morphology of the male genitalia are illustrated in Fig. 3 AC. Females of these two species can be distinguished based on the shape of the emargination of the distal edge of the 8th abdominal sternite. The emargination is deep, i.e., two times deeper than its maximal width, in B. kukalovae n. sp. In B. weidneri , in contrast, the emargination is as wide as deep and has a semicircular shape. Brazilozoros huxleyi is a visually conspicuous species, easily recognized by the hind legs, which bear three conspicuous spurs on the ventral edge in the case of B. huxleyi . In clear contrast only 10 short bristles are present in B. kukalovae n. sp. This character applies to both sexes. Brazilozoros brasiliensis is known only from the female sex, with the exception of only one described gynandromorphic specimen ( Rafael et al. 2017 ). Based on this specimen, we suspect that the anterior group of sensilla basiconica on the 10th tergite of males is composed of two sensilla (which are not divided basally) in the case of B. brasiliensis but of four sensilla in the case of B. kukalovae n. sp. Females of the two species differ in the emargination of the distal edge of the 8th abdominal sternite, which is shallow, broadly concave, and wider than deep in B. brasiliensis vs. two times deeper than wide in B. kukalovae n. sp. All described species of Brazilozoros can be identified with the aid of the key provided in this study. ETYMOLOGY. — This species is named after Dr Jarmila KukalovaPeck in honour of her contribution to Zoraptera research.lishein GenBank as B. weidneri ( New, 1978 ) (see Matsumura et al. 2020 ). Abbreviation: N/A , not available. TABLE 1. — Details of the material used in the phylogenetic analyses. Newly obtained sequences are marked in bold . Asterisk ( * ) indicates the specimens pub-

Analysed taxa	Sampling locality	Isolate	H3	16S	18S
Zorotypidae Silvestri, 1913
Spermozorinae Kočárek, Horká & Kundrata, 2020
Spermozoros asymmetricus (Kočárek, 2017)	Brunei	4Z	ON811689	MN790592	ON807040
Zorotypinae Silvestri, 1913
Zorotypus delamarei Paulian,1949	Madagascar	71Z	ON811690	MN790583	ON807041
Usazoros hubbardi (Caudell,1918)	USA: Florida	89Z	ON811691	ON722350	ON807042
Spiralizoridae Kočárek, Horká & Kundrata, 2020
Latinozorinae Kočárek, Horká & Kundrata, 2020
Latinozoros cacaoensis Kočárek & Horká, 2023	French Guiana	48Z	ON811693	ON722349	ON807044
Latinozoros gimmeli Kočárek & Horká, 2023	Dominican Republic	41Z	ON811692	ON722348	ON807043
Spiralizorinae Kočárek, Horká & Kundrata, 2020
Brazilozoros brasiliensis ( Silvestri, 1946 )	Brazil: Paraná	YK29	LC477128	LC476748	LC477104
Brazilozoros “ huxleyi ” (Bolívar y Pieltain & Coronado, 1963) Ecuador		YK11	LC477129	LC476749	LC477105
Brazilozoros “ huxleyi ”	Peru	YK20	LC477131	LC476751	LC477106
Brazilozoros “ huxleyi ”	Peru	YK21	LC477132	LC476752	LC477107
Brazilozoros kukalovae n. sp.	French Guiana	94Z	N/A	OP562643	OP558030
Brazilozoros kukalovae n. sp.	French Guiana	YKFG	LC477127*	LC476747*	LC477103*
Brazilozoros kukalovae n. sp.	Brazil: Ubajara	YK18	LC477126*	LC476746*	N/A
Brazilozoros sp.	French Guiana	93Z	N/A	OP562642	OP558031
Brazilozoros ” weidneri ” ( New,1978 )	Brazil: Amazonas	YK7	LC477124	LC476744	LC477101
Brazilozoros “ weidneri ”	Brazil: Roraima	YK17	LC477125	LC476745	LC477102
Spiralizoros cervicornis ( Mashimo, Yoshizawa & Engel, 2013 ) Brunei		1Z	ON811694	MN790588	ON807046
Spiralizoros magnicaudelli (Mashimo, Engel, Dallai, Beutel	Malaysia: Sabah	6Z	ON811695	MN790600	ON807047
& Machida, 2013)
Outgroup (Polyneoptera)
Blattodea : Gromphadorhina portentosa	N/A	B01; N/A	AY125216	Z97626	Z97592
Dermaptera: Euborellia arcanum	USA: Florida	E8	ON856150	MN790595	MN790614
Grylloblattodea: Grylloblatta campodeiformis	USA: MT	BYU_gb31.1	DQ457398	DQ457262	DQ457299
Mantodea: Mantis religiosa	Ghana	MN247	FJ806794	FJ806243	FJ806432

DISTRIBUTION, HABITAT, AND BIOLOGY. — Brazilozoros kukalovae n. sp. was collected from under the bark of rotting logs in a lowland rainforest in French Guiana at altitudes from 25 to 40 m a.s.l. ( Fig. 5 ). The species is currently known only from French Guiana , but we expect its occurrence in similar habitats in neighbouring countries in Amazonia. DESCRIPTION Apterous male Measurements. Total body length: 2.58-2.71 mm , head width 0.52-0.54 mm , head length 0.40-0.41, antenna length 1.42- 1.44 mm , pronotal width 0.44-0.45 mm , metafemur length 0.65-0.66 mm , metatibia length 0.60, abdomen maximal width 0.55-0.57 mm , cerci length 0.11 mm . Colour. Body color pale brown; tibiae and tarsi of all legs, cerci, and membranous regions lighter; antennae pale brown with antennomeres VII-IX lighter ( Fig. 5E ). Head. Head subtriangular, about as long as broad posteriorly, and triangular in dorsal view, slightly wider than pronotum ( Fig. 1 A-C); coronal and frontal sutures invisible; y-shaped ecdysial cleavage lighter ( Fig. 1 A-C); cephalic setae ( Fig. 1B ) short and sparse, not grouped; compound eyes and ocelli absent; fontanelle (cephalic gland) absent; antennae 9-segmented ( Fig. 1A, D ), antennomeres II-VI with several long setae (longer than diameter of antennomere), antennomere I slightly curved outward, antennomere II short, about half the length of antennomere I, antennomere III slightly longer than antennomere II; antennomeres IV-VIII longer than wide, distally narrowed, with maximal with in distal third, antennomere IX cylindrical, distally narrowed. Mandibles asymmetrical, triangular with rounded lateral margins, each with five teeth on distal half and serrated molar region; left mandible with apically branched stick-like prostheca, inserted on ventral surface of second (subapical) tooth; maxillary palpus five-segmented; labial palpus threesegmented ( Fig. 1C ). Thorax. Pronotum subrectangular, wider than long, slightly narrowed posteriorly and setose, chaetotaxy as depicted in Fig. 1B ; median ecdysial suture lighter; prosternum with peglike lateral protuberance proximally on each side ( Fig. 1C ); mesonotum and metanotum trapezoidal, distinctly wider than long, shorter than pronotum. Legs. Covered by short setae ( Fig. 1A, E ); distal half of protibia with row of flat sensilla ventrally, as long as setae on tibia; metafemur broad with maximal width in middle part and gradually tapering towards apex ( Fig. 1E ); dorsal surface broadly setose, ventral surface with 10 bristles; length of bristles I, III, and V-X similar to length of dorsal setae, bristle I and IV distinctly shorter and thinner ( Fig. 1E ); metatibia with densely arranged setae and two strongly sclerotized spurs ventrally of similar length as setae in distal third of tibia; distal end of tarsomere II with short unguitractor plate and hooked paired claws ( Fig. 1E ). FIG. 1. — Brazilozoros kukalovae n. sp. , apterous male and female paratypes: A , dorsal view of male; B , head and pronotum of male from dorsal view; C , head and pronotum of male from ventral view; D , right antenna of male; E , right hind leg of male, anterior view; F , right hind leg of female, anterior view. Abbreviations: bI-bX , bristles; cl , clavus; cox, coxa; ecc , ecdysial cleavage; lbp , labial palp; lpp , lateral protuberance of prosternum; mdb , mandibula; mxl , maxilla; mxp , maxillary palp; mtf , metafemur; mtt , metatibia; prn , pronotum; prs , prosternum; ta , tarsus. Scale bars: A, 1.0 mm; B, C, 0.2 mm. Abdominal tergites. I to VII (T1-7) with single transverse row of four to six setae of moderate length distally and few lateral, short setae on each side of midline ( Fig. 2A, F); T8 with two setae posteriorly on each side of midline ( Fig. 2A ); T9 short, with anterior trapezoidal expansion extending beneath T8, with single transverse row of two shorter and three longer setae posteriorly on each side of midline ( Fig. 2A, F ); T10 posteromedially incised, with three short setae on each side of midline and an anterior and posterior group of thick, black sensilla basiconica; anterior group composed of four sensilla basiconica, posterior group of two sensilla basiconica ( Fig. 2A , E-G); T11 with median upcurved projection (= male mating hook) and two lateral, subtriangular sclerites, covered by several short and moderate-length setae on each sclerite ( Fig. 2D ); epiproct and paraproct unsclerotized; cercus unsegmented, oval, with several moderate-length setae and several very long, fine setae ( Fig. 2A, B , D-G); surface covered with numerous minute spicules except at base and apex ( Fig. 2G ); sternum I (S1) scarcely sclerotized; S2-S3 weakly sclerotized with single transverse row of several moderate-length setae distally; S4-6 with single transverse row of four to six moderate-length setae on each side of midline ( Fig. 2A, F ); S7 covered by moderate-length setae; posterior edge with 3-4 longer seta on each side of midline ( Fig. 2B, D ); S8 strongly transverse, with row of short setae anteriorly and row of several (3-4) moderate-length setae posteriorly on each side of midline ( Fig. 2A ); S9 transverse, rounded, distally with small, fine setae ( Fig. 2B ); S10 invaginated beneath S9, weakly sclerotized, excised distally and not visible externally in living specimens – sclerite partly visible after maceration on Fig. 2G; S 11 weakly sclerotized, composed of two lateral sclerites, each with small setae ( Fig. 2B, D ). FIG. 2. — Brazilozoros kukalovae n. sp. , apterous male and female paratypes: A , posterior segments of male abdomen, dorsal view; B , posterior segments of male abdomen, ventral view; C , posterior segments of female abdomen, ventral view; D , tip of male abdomen, posterior view; E , tip of female abdomen, posterior view; F , male abdomen in lateral view, dark field; G , latero-dorsal view of tip of male abdomen, dark field. Abbreviations: ce , cercus; mp , median up-curved projection; S , abdominal sternite; sba , anterior group of sensilla basiconica; sbp , posterior group of sensilla basiconica; T , abdominal tergite; I-VI , sclerites of male genital. Scale bars: 0.2 mm. FIG. 3. — Brazilozoros kukalovae n. sp. , apterous male paratype ( A , B , D ) and B. weidneri ( New,1978 ),apterous male paratype ( C , E ): A , ventral view of male genitalia of B. kukalovae n. sp. ; B , anterior view of sclerites VIa,b of male genitalia in B. kukalovae n. sp. ; C , ventral view of male genital of B. weidneri ; D , tip of male abdomen in B. kukalovae n. sp. , posterior view; E , tip of male abdomen in B.weidneri , posterior view.Abbreviations: ce , cercus; mp , median up-curved projection; sba , anterior group of sensilla basiconica; sbp , posterior group of sensilla basiconica; T , abdominal tergite; I-VI , sclerites of male genitalia. Scale bars: 0.2 mm. Male genitalia. Symmetrical ( Fig. 3A, B ). Composed of antero-posteriorly oriented basal plate (sclerite I) with long, anterior tongue-like process, and bifurcated posterior part; antero-posteriorly oriented pair of central ventral sclerites (sclerite IIa,b) attached to basal plate, with laterally oriented spines anteriorly, pointed and hooked (mutually divergent) distal ends, each with minute teeth on distal third, oriented laterally; pair of sinuous transverse sclerites attached to basal plate (sclerite IIIa,b), each with elongate outer projection; pair of transverse arms (sclerites IVa,b) gradually narrowing from base to apex and curved symmetrically towards sclerites III; medial part rugose, anteriorly two rugose lobes inserted on each arm, one smaller and rounded, second larger and truncated anteriorly; central part of genital apparatus bordered by narrow lateral sclerites (sclerites Va,b); dorsally placed pair of needle-like sclerites (sclerites VIa,b) not attached to rest of genital apparatus, sclerites oriented antero-posteriorly ( Fig. 2A, E ). FIG. 4. — Brazilozoros kukalovae n. sp. , dealate female paratype and wings of alate female: A , dorsal view of head and thorax; B , forewing, dorsal view; C , hind wing,dorsal view. Abbreviations: Cu , cubitus vein; CuA , anterior cubitus; ey , compound eye; M , media vein; msn , mesonotum; mtn , metanotum; oc , ocelli; prn , pronotum; R (Rs) , radius vein. Scale bars: 0.5 mm. Apterous female Morphology and measurements. Similar to male. Except for following features: metafemur slender, ventral surface with the same arrangement of spines as in males, but spines thinner ( Fig. 1F ); abdomen wider, maximum width 0.61-0.69 mm ; T10 transverse, not incised posteromedially, with row of small scattered setae on proximal and distal edge; T11 trapezoidal, not divided into two halves (hemitergites), covered with small scattered setae and pair of paramedian setae of moderate length apically, distal edge weakly incised; S8 strongly trapezoidal, wider than long, with short setae evenly scattered and longer setae flanking the distal and lateral edges; distal end deeply triangularly excised ( Fig. 2C ); S9 transverse, mostly covered by S8, distal edge medially with triangular protuberance juxtaposed to excision of S8 basally (gonopore between excision of S8 and protuberance of S9); S10 entire, not divided into hemitergites, transverse, weakly emarginate anteriorly, with row of small scattered setae on proximal and distal edge. Description of alate/dealate specimens Morphology. Generally similar to those of the apterous specimens except as follows: blackish brown coloration; compound eyes and three black ocelli present, ocelli arranged as triangle, two anterior to compound eyes, and one medially on anterior frons; distal edge of pronotum only weakly sclerotized; mesonotum and metanotum indistinctly divided into slightly pointed prescutum, large scutum, and smaller and posteriorly rounded scutellum ( Fig. 4A ). Wings. Brown-hyaline with dense pubescence ( Fig. 4B, C ). Forewing 2.3 mm long, hind wing 1.8 mm . Forewing veins ( Fig.4B ) M and Rs weak and indistinct; pterostigma present on anterior margin between R and C veins; Rs far from the wing tip, reaches C vein close to pterostigma; veins rs-m present, slightly shorter than abscissa of Rs; M thin, extended to posterior margin; M + Cu coarse and nebulous; CuA 1 extending from posterior wing margin; CuA 2 short, almost absent. Hind wing ( Fig. 4C ) with long R+M, R and M 1+2 veins well defined, M vein weaker; R and M 1+2 fused in distal third of wing; M short and indistinct, starting near the base of the wing; Cu absent. MOLECULAR IDENTIFICATION FIG. 5. — Habitat and living specimens of Brazilozoros kukalovae n. sp. : A , rotting logs in type locality, Montagne des Singes (French Guiana: around Kourou); B , molokoï track (French Guiana: around Cacao); C , collecting of Zoraptera in Montagne des Singes (above Marek Jankásek, below Petr Kočárek); D , nymph of B. kukalovae n. sp. with a small unidentified isopod (above) from the family Philosciidae Kinahan, 1857 , commonly occurring together with zorapterans in French Guiana; E , apterous female. Photographs: P. Kočárek (A, B, D, E) and I. H. Tuf (C). We obtained a partial 16S rRNA sequence (500 bp) of Brazilozoros kukalovae n. sp. as a DNA barcode for the purpose of molecular identification of the species; the sequence has been deposited in GenBank under accession number OP562643 (isolate number: 94Z). TABLE 2. — Genetic divergences of Brazilozoros Kukalova-Peck & Peck, 1993 species were analysed using the Kimura 2-parameter model. Molecular markers 16S (302 analysed positions) and 18S (459 analysed positions) are shown; values of 16S are below the diagonal. Symbol: * , specimens published in GenBank as B. weidneri ( New, 1978 ) (see Matsumura et al. 2020 ); N/A , not available.

1	2	3	4	5	6	7	8	9	10
1	B. kukalovae n. sp. 94Z Fr. Guiana	0.00	N/A	0.00	0.00	0.02	0.02	0.03	0.02	0.03
2	B. kukalovae n. sp. YKFG Fr. Guiana *	0.00	N/A	0.00	0.00	0.02	0.02	0.03	0.02	0.03
3	B. kukalovae n. sp. YK18 Brazil *	0.04	0.04	N/A	N/A	N/A	N/A	N/A	N/A	N/A
4	B. “ weidneri ” YK7 Brazil	0.16	0.16	0.18	0.00	0.02	0.02	0.03	0.02	0.03
5	B. “ weidneri ” YK17 Brazil	0.17	0.17	0.20	0.03	0.02	0.02	0.03	0.02	0.03
6	B. sp. 93Z Fr. Guiana	0.23	0.23	0.25	0.24	0.25	0.00	0.01	0.00	0.01
7	B. “ huxleyi ” YK20 Peru	0.26	0.26	0.27	0.26	0.28	0.05	0.01	0.00	0.02
8	B. “ huxleyi ” YK11 Ecuador	0.23	0.23	0.26	0.26	0.27	0.07	0.07	0.01	0.02
9	B. “ huxleyi ” YK21 Peru	0.25	0.25	0.26	0.25	0.27	0.07	0.06	0.07	0.01
10	B. brasiliensis YK 29 Brazil	0.19	0.19	0.21	0.22	0.22	0.27	0.27	0.25	0.26

KEY TO SPECIES OF BRAZILOZOROS KUKALOVA- PECK & PECK, 1993 1. Male ............................................................................................................................................................ 2 — Female ........................................................................................................................................................ 5 2. 10th tergite with two sensilla basiconica (one anterior, one posterior) on each side of midline. Metafemur with three conspicuous spurs on ventral edge, much longer than diameter of tibia ................................................. ............................................................................ Brazilozoros huxleyi (Bolivar y Pieltain & Coronado, 1963) — 10th tergite with two groups of sensilla basiconica on 10th tergite, each composed of at least two sensilla. Metafemur with number of bristles, length of bristles is less then diameter of tibia ................................................ 3 3. Anterior group of sensilla basiconica composed of 4 sensilla, anterior sensilla shorter than sensilla in posterior group ................................................................................................................. Brazilozoros kukalovae n. sp. — Anterior group of sensilla basiconica composed of 2(3) sensilla, anterior sensilla longer than sensilla in posterior group .......................................................................................................................................................... 4 4. Lateral sensilla basiconica of anterior group divided basally (to two protrusions of similar length) .................. .................................................................................................................. Brazilozoros weidneri ( New, 1978 ) — Anterior group of sensilla basiconica composed of two sensilla, not-divided basally (based on only one known gynandromorph male – see Rafael et al. 2017 ) .................................. Brazilozoros brasiliensis ( Silvestri, 1946 ) 5. Metafemur with number of bristles, length of bristles is less then diameter of tibia ...................................... 6 — Metafemur with three conspicuous spurs on ventral edge, as long as or longer than diameter of tibia ............. ........................................................................................................................................................ B. huxleyi 6. Distal emargination of 8th sternite deep, two times deeper than its maximal width ............ B. kukalovae n. sp. — Distal emargination of 8 th sternite wider, shallow ........................................................................................ 7 7. Distal emargination of 8th sternite semicircular, as deep as wide ................................................... B. weidneri — Sternite 8 with shallow, broadly concave emargination, significantly wider than deep ................ B. brasiliensis PHYLOGENETIC POSITION OF BRAZILOZOROS KUKALOVAE N. SP. The final analysed dataset of the three markers (18S, 16S, and H3) consisted of 1,106 bp. The phylogenetic tree based on the ML method ( Fig. 6 ) shows a strongly supported sister position of the new species B. kukalovae n. sp. with B. “ weidneri ” . The genetic divergences within Brazilozoros species based on 16S and 18S rRNA sequences using the Kimura 2-parameter model are presented in Table 2 . The maximum divergence of B. kukalovae n. sp. ( Fig. 6 , clade 1) to B. “ weidneri ” (clade 2) is 20% for 16S and 0% for 18S. The maximum value between specimens of B. kukalovae n. sp. from French Guiana and specimens from Brazil is 4% for 16S; the 18S sequence from Brazil was not available.