(Isopoda: Cymothoidae), with Description of Six New Species of Anilocra Leach, 1818 from Africa Rachel L

Welicky and Smit Parasites Vectors (2019) 12:387 https://doi.org/10.1186/s13071-019-3578-5 Parasites & Vectors

RESEARCH Open Access Redescription and molecular characterisation of the fsh ectoparasite Anilocra capensis Leach, 1818 (Isopoda: Cymothoidae), with description of six new species of Anilocra Leach, 1818 from Africa Rachel L. Welicky1,2* and Nico J. Smit1

Abstract Background: Anilocra capensis Leach, 1818 is the only named species of Anilocra Leach, 1818 from South Africa. Anilocra is a large genus (> 40 species) with high levels of diversity reported from the Caribbean and Indo-West Pacifc. Considering it is highly unlikely that all records of Anilocra from South Africa can be of a single species, the aim of this study was to better understand the diversity of Anilocra from this region and continent. Methods: To redescribe A. capensis, the syntypes of A. capensis and specimens recorded as A. capensis from Africa were borrowed from the Natural History Museum, London, UK, and The iZiko South African Museum, Cape Town. Newly collected fresh samples of A. capensis were collected from of Cape Town, South Africa. Morphological rede- scriptions of the syntypes, and other museum and fresh material were conducted. Fresh samples were used to charac- terise molecularly A. capensis using the mitochondrial cytochrome c oxidase subunit 1 gene (cox1). Results: Morphological analyses demonstrated that apart from A. capensis there are six Anilocra species new to science from Africa: Anilocra ianhudsoni n. sp., Anilocra bunkleywilliamsae n. sp., Anilocra paulsikkeli n. sp., Anilocra jovanasi n. sp., Anilocra angeladaviesae n. sp. and Anilocra hadfeldae n. sp. Of the species under study, specimens of A. capensis appear to demonstrate the most individual variation, which occurs in pleonite width, pleotelson form and uropod length. We determined that African species of Anilocra can be primarily diferentiated by the proportional shape and size of the full body in dorsal view and pereonites 1, 6 and 7. Other defning morphological traits include proportional shape and size of the pereopods, and the antenna and antennula peduncles. Lastly, the molecular characterisation of A. capensis is provided and the interspecifc divergence with Mediterranean species is smaller than that with Carib- bean species. Conclusions: The results of this study provide a detailed redescription of A. capensis and the frst molecular barcode for this organism. Six new species of Anilocra from Africa are described, establishing that the diversity of Anilocra in this region is greater than previously known. With this new understanding of species diferences, we can accurately conduct detailed molecular and ecological analyses of Anilocra from Africa with certainty of the organism under study. Keywords: Africa, Anilocra, Cymothoid, Fish parasite, Molecular characterization, Morphological description

*Correspondence: [email protected] 2 Present Address: School of Aquatic and Fishery Sciences, University of Washington, 1122 NE Boat Street, Seattle, WA 98105, USA Full list of author information is available at the end of the article

© The Author(s) 2019. This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/ publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated. Welicky and Smit Parasites Vectors (2019) 12:387 Page 2 of 34

Background Oceans. Moreover, we aim to provide molecular data for Members of the isopod fsh parasitic family Cymothoidae A. capensis so that future taxonomic studies can at a min- Leach, 1818 conspicuously infest the skin, buccal cavity, imum describe Anilocra from Africa as A. capensis or a or gill chamber of their hosts. Tis family has a global dis- diferent species of Anilocra. tribution and there are more than 43 recognised genera [1]. One of the most commonly observed genera is Ani- Methods locra Leach, 1818, and it remains one of the best studied Specimen collection ecologically [2–4]. In fact, Anilocra was the frst genus of In March 2017, specimens identifed as Anilocra capen- a marine fsh parasite to be described from South Africa sis with locality data from Africa, along with the syntypes [5]. of A. capensis were borrowed from the Natural History Leach [6] erected Anilocra to accommodate three spe- Museum, London, UK (NHM). We examined other spec- cies. One of these species was Anilocra capensis Leach, imens of Anilocra from outside South Africa from NHM 1818, collected in the Cape region of South Africa. In to confrm or reject if they were A. capensis. In July 2017, Schioedte & Meinert’s (1879) redescription of A. capen- all specimens labelled as Anilocra from Te iZiko South sis, the authors provided only two dorsal view drawings African Museum, Cape Town (SAM) were borrowed. In of the female, and one dorsal view drawing of the male April 2018, live A. capensis were collected from the Hot- [7]. Schioedte & Meinert (1879) did not examine Leach’s tentot seabream, Pachymetopon blochii Valenciennes, type-material. Since 1879, no further supplemental rede- at two localities: Cape Town Harbour wall and the False scription of A. capensis has been published. As a result, Bay Yacht Club harbour. Infested fshes were speared nearly all reports of Anilocra infestation on South Afri- on SCUBA under the direction and permitting of Two can marine fshes have been recorded as A. capensis. Oceans Aquarium. Te fresh material was preserved in Te majority of Anilocra have been reported from 80% ethanol for later morphological analysis (see Table 1 tropical environments, e.g. [8–10]. Although cymothoids for sample sizes and locality data). For some specimens, are generally more speciose in tropical than in temper- leg tissue was extracted using forceps and placed in 96% ate environments, temperate systems are nonetheless ethanol for molecular analysis. diverse. Accordingly, Anilocra diversity in South Africa should be far greater than the single recorded species. Molecular analysis Tis should particularly be the case as South Africa’s Genomic DNA was extracted from 8 of the freshly col- coastline encompasses two oceans that span temperate to lected A. capensis. A rapid DNA extraction method as subtropical ecosystems. Moreover, these ecosystems are described in the KAPA Express Extract Kit (Kapa Biosys- suitable for a diversity of potential fsh hosts. tems, Cape Town, South Africa) was used. Polymerase To test the aforementioned prediction, a review of A. chain reactions (PCR) were used to amplify a 710 bp frag- capensis based on the syntypes and all other available ment of the mitochondrial cytochrome c oxidase subunit material was warranted. We seek to provide descriptions gene (cox1) using the primer sets LCO 1490 and HCO of species of Anilocra from South Africa and the African 2198 [11]. PCR was performed using 12.5 μl Termo Sci- continent in order to clarify and describe the diversity of entifc DreamTaq PCR master mix (2×) (2× DreamTaq Anilocra from the eastern Atlantic and western Indian bufer, 0.4 mM of each dNTP, and 4 mM MgCl2), 1.25 μl

Table 1 Details of Anilocra species examined Collection locality Male Female

Anilocra capensis Of Cape Town (Harbor Wall), South Africa 8 9 False Bay, Cape Town, South Africa 26 38 Table Bay, Cape Town, South Africa 7 Cape Town region, South Africa 5 Anilocra ianhudsoni n. sp. Nahoon River, East London, South Africa 1 2 Anilocra bunkleywilliamsae n. sp. Mawalana Estuary, South Africa 1 2 Anilocra paulsikkeli n. sp. Delagoa Bay, Mozambique 2a 2 Anilocra jovanasi n. sp. Delagoa Bay, Mozambique 1 Anilocra angeladaviesae n. sp. Of Cape Blanc, Morocco 2 4 Anilocra hadfeldae n. sp. Of Cape Blanco, Gambia 1 1 a One specimen may be transitional Welicky and Smit Parasites Vectors (2019) 12:387 Page 3 of 34

of each primer, 1 μl DNA, and 9 μl of PCR-grade nucle- order classifcation follows Brandt & Poore [18]. Fish ase free water (Termo Fisher Scientifc, Vilnius, Lithu- nomenclature follows FishBase [19]. ania). Total volume per reaction was 25 μl, and PCR To comply with the regulations set out in article 8.5 of reactions were conducted using a ProFlex™ PCR thermal the amended 2012 version of the International Code of cycler (Applied Biosystems by Life Technologies). Reac- Zoological Nomenclature (ICZN [20], details of all new tions were amplifed under the following PCR conditions: taxa have been submitted to ZooBank. For each new Stage 1, 94 °C for 5 min, Stage 2, 36 cycles of 94 °C for 30 taxon, the Life Science Identifer (LSID) is reported in the s, 47 °C for 50 s, 72 °C for 2 min, and Stage 3, 72 °C for 10 taxonomic summary. min. PCR products were sent to a commercial sequencing company (Inqaba Biotechnical Industries (Pty) Ltd, Results Pretoria, South Africa) for purifcation and sequencing Specimen summary in both directions. Obtained sequences were assembled, We identifed from the museum loans and feld collection and chromatogram-based contigs were generated using A. capensis and six new species of Anilocra. Tese speci- Geneious Ver. 9.1. Sequences were aligned and trimmed mens can be distinguished using the key provided here. to the length of the shortest sequence using MEGA 7 Sample sizes and locality data for each species are given software program [12]. in Table 1. Using BLASTn (Basic Local Alignment Search Tool; http://www.ncbi.nlm. nih.gov/blast), the obtained Molecular analyses sequences were verifed as belonging to the Isopoda. Comparative sequence analysis indicated that the Mean intraspecifc divergence for A. capensis and inter- intraspecifc divergence of A. capensis was 1–7 nt specifc divergence with all other Anilocra reported on (0.04%). Te interspecifc divergence between A. capensis GenBank were examined using p-distance and number and A. physodes was 63–74 nt (15.5%). Te interspecifc of bp diferences. We utilised the following sequences: divergence between A. capensis and Caribbean species of Anilocra physodes Linneaus, 1758 (EF455817); Anilo- Anilocra were: A. chromis: 138–157 nt (24.9%), A. brillae: cra chromis Williams & Williams, 1981 (KY5622739); 138–157 nt (24.9%), A. haemuli: (KY562752) 136–154 Anilocra brillae Welicky, Hadfeld, Sikkel & Smit, 2017 (24.4%), A. prionuri: (LC159541) 136–157 (24.8%), and A. (KY562744); Anilocra haemuli Williams & Williams, clupei: (LC160309) 132–144 (24.4%). 1981 (KY562752); Anilocra prionuri Williams & Bunkley- Williams, 1986 (LC15954); and Anilocra clupei Williams Taxonomy & Bunkley-Williams, 1986 (LC160309). Newly-generated sequences for Anilocra capensis were deposited in the GenBank database under the accession numbers Genus Anilocra Leach, 1818 MK450443-MK450452. Diagnosis. Diagnosis and synonymy in Bruce [21]. Body dorsally symmetric, weakly to strongly vaulted. Morphological data Cephalon non-tri-lobed to weakly tri-lobed, anterior of Specimens of Anilocra were processed according to the frontal margin of rostrum blunt and folded down poste- techniques described in [13, 14]. Dissection was permit- riorly between antennule bases. Antennula shorter than ted for specimens from SAM but not the NHM. Regard- antenna. Pereonite 5 or 6 widest, coxae 4–6 longer, more ing the museum collection specimens in this study, SAM anteriorly pointed than 1–3. Pereopods 1–7 increase in assigns one accession number per species and collection size. Type-species: Anilocra cuvieri Leach, 1818. location, whereas NHM assigned one accession number per individual in most cases. Accession numbers Remarks from NHM are indicated with the abbreviation BMNH. Leach [6] described three species: Anilocra cuvieri Leach, Descriptions were prepared using DELTA (Descriptive 1818, Anilocra mediterranea Leach, 1818 and Anilocra Language for Taxonomy) [15] using a general character capensis. Anilocra cuvieri was designated as the type-spe- set for the Cymothoidae [16, 17]. Ratios and measure- cies by Kussakin [22]. Tere are three Anilocra capensis ments were rounded of to one decimal place and were syntypic specimens held at Te Natural History Museum, made using maximum values of the specifc measured London, presented by Leach from the Cape of Good article. Measurements were taken and ratios determined Hope (see ‘Material examined’) [23]. from ovigerous and non-ovigerous female (♀), and male Nearly half of the species of Anilocra accepted today (♂) specimens. Pleotelson length (TL) and width (W) were described in the 1980s, with many originating from for all specimens examined are reported as TL × W. All the Indo-Pacifc and Caribbean [8, 21]. Since 2001, just measurements are reported in millimeters (mm). Higher four new species have been described based on material Welicky and Smit Parasites Vectors (2019) 12:387 Page 4 of 34

from the Caribbean [24], Chile [25], Lebanon [26] and 50.5 × 25.0), damaged ♀ (27.0 × 9.5; 29.5 × 12.0; the Gulf of Mexico [27]. In 2017, using morphological 32.0 × 13; 37.0 × 14.0; 37.0 × 14.0; 37.0 × 17.0; 37.5 × 14.0; and molecular data, Anilocra brillae [24] from the Car- 40.0 × 18.0). All specimens collected by R. L. Welicky, O. ibbean was identifed and described based on specimens Kudlai, A. Greyling, D. van Rooyen, T. Beukes and Two that were originally thought to be Anilocra haemuli [8]. Oceans Aquarium personnel on 28–29 April 2017 on the Tis revision is an example of the cryptic nature of some host P. blochii. Anilocra spp., and the importance of combining morpho- Additional museum material examined: Collected logical and molecular data whenever possible. by UCT Ecological Survey from Table Bay on host P. Of the skin-attaching cymothoids, species of Anilocra blochii and held at SAM as SAMA44882: ♀ (44.0 × 18.0; most closely resemble species of Renocila Miers, 1880 44.0 × 20.0; 47.0 × 19.0; 47.0 × 21.0; 49.0 × 21.0; and Nerocila Leach, 1818, but generic level diferences 50.0 × 22.0; 53.0 × 22.0). Collected from SAM and not can be distinguished without dissection or stereoscopic catalogued: ♀ (45.0 × 22.0; 49.0 × 19.0; 49.0 × 20.0; microscope [28–30]. Briefy, in Anilocra spp. pereopod 50.0 × 20.0; 50.0 × 22.0). 7 is longer than 6, but in Renocila spp. pereopod 6 is Representative DNA sequences: MK450443-MK450452 similar in length to 7. Te rostrum is not ventrally folded between the antennae in Renocila spp., but it is in Ani- Description locra spp. Te cephalon of Nerocila spp. is posteriorly Female lectotype. [BMNH 1979.329:207b; Fig. 1.] Size strongly trilobed, whereas the cephalon of Anilocra spp. 43.0 × 18.0. Body ovoid, 2.6 times as long as greatest is not tri-lobed to very weakly tri-lobed. Further compar- width; dorsal surfaces smooth and polished in appear- isons among cymothoid genera and generic features are ance; widest at pereonite 5, most narrow at pereonite detailed in Welicky et al. [24]. 1; pereonite lateral margins mostly posteriorly ovate. Cephalon 0.6 as long as wide as measured in dorsal view, Anilocra capensis Leach, 1818 trapezoid-shaped. Pereonite 1 smooth, anterior border straight, anterolateral angle narrowly rounded, in line Type-host: Probable host Pachymetopon blochii referred with base of cephalon. Posterior margins of pereonites to as “Sargus Hottentotus” in Schiöedte & Meinert (1883) smooth, slightly curved laterally. Coxae 2–3 wide, with [7]. posteroventral angles rounded; coxae 4–7 with rounded Type-material: BMNH 1979.329:207b (♀ 43.0 × 18.0) point, curved, not extending past posterior pereonite (designated as the lectotype in the present study); BMNH margin; coxa 7 extending past anterior pereonite margin, 1979.329:207c (♀ 46.0 × 19.5); BMNH 1979.329:207a (♂ coxae 4–6 not extending past anterior pereonite margin. 35.0 × 10.0). Paralectotypes: BMNH 1979.329:207c (♀ Pereonites 1–5 increasing in length and width; pere- 46.0 × 19.5), BMNH 1979.329:207a (♂ 35.0 × 10.0). Pre- onites 6–7 decreasing in length and width; pereonites 5 sented by W. Leach in 1818 from Cape of Good Hope. and 6 subequal, or pereonites 1–4 narrower. Pleon with New material examined: From Cape Town Harbor Wall pleonite 5 nearly widest, visible in dorsal view; posterior ′ ″ ′ ″ (33°53 58 S, 18°25 35 E) SAMC-A091297: ♂ (22.0 × 7.0; margin of pleonites 1–4 posteriorly concave, pleonite 5 25.0 × 7.0; 26.0 × 8.5; 27.0 × 8.0; 27.0 × 8.5; 29.0 × 9.0; posteriorly slightly concave to straight or smooth, mostly 31.5 × 9.0; 33.0 × 10.0), non-ovigerous ♀ (43.5 × 16.5; concave. Pleonite 2 not laterally overlapped by pereonite 44.5 × 20.0), ovigerous ♀ (43.0 × 18.5; 44.0 × 19.5; 7; posterolateral angles of pleonite 2 narrowly rounded. 45.0 × 19.5; 45.0 × 19.5; 49.0 × 20.0; 49.0 × 20.0), dam- Pleonite 1 similar in form to pleonite 2 and 3. Pleonites aged ♀ (46.6 × 19.5). From False Bay Yacht Club Marina 3–5 similar in form to pleonite 2; pleonite 5 widest or ′ ″ ′ ″ (34°11 32 S, 18°26 01 E) SAMC-A091298: ♂ (12.0 × 3.5; nearly widest to pleonite 4, not overlapped by lateral 15.0 × 4.0; 18.0 × 5.0; 18.5 × 5.0; 19.0 × 4.5; 19.0 × 5.0; margins of pleonite 4 or with posterolateral angles nar- 19.5 × 5.5; 20.0 × 5.0; 21.0 × 5.5; 21.0 × 5.5; 21.0 × 6.0; rowly rounded, posterior margin straight. Pleotelson 0.8 22.0 × 6.0; 23.0 × 6.0; 23.0 × 6.5; 23.0 × 7.0; 23.0 × 8.0; times as long as anterior width, dorsal surface smooth; 23.6 × 6.5; 24.0 × 5.0; 24.0 × 6.5; 24.0 × 7.0; 24.0 × 7.0; lateral margins convex, posterior margin evenly rounded. 25.0 × 7.0; 28.0 × 8.0; 28.0 × 8.0; 28.5 × 8.0; 29.0 × 9.0), Antennula approximately as stout as antenna, com- non-ovigerous ♀ (26.0 × 10.0; 27.0 × 9.5; 27.5 × 9.5; prised of 8 articles; peduncle articles 1 and 2 distinct, 29.5 × 9.0; 29.5 × 9.0; 34.0 × 13.0; 35.5 × 14.0; 39.0 × 13.0; articulated. Antenna comprised of 8 articles. 42 × 18; 43.5 × 16.5; 44.5 × 20.5; 51.5 × 23.0), oviger- Uropod more than half the length of pleotelson, pedun- ous ♀ (28.0 × 12.0; 29.0 × 11.0; 34.5 × 13.0; 34.5 × 15.0; cle 0.4 times as long as rami, peduncle lateral margin 36.0 × 16.0; 37.5 × 15.0; 38.0 × 15.0; 38.0 × 15.5; without setae or without medial short acute robust seta; 38.0 × 15.5; 39.0 × 14.0; 40.0 × 15.0; 40.0 × 15.0; rami extending beyond pleotelson, marginal setae absent, 41.0 × 15.0; 43.0 × 18.5; 43.5 × 17.0; 47.0 × 19.0; apices broadly rounded. Endopod apically rounded, 3.6 Welicky and Smit Parasites Vectors (2019) 12:387 Page 5 of 34

a b d

g f i

Fig. 1 Anilocra capensis (a–e 43.0 18.0, BMNH 1979.329:207b; f–j 46.0 19.5, BMNH 1979.329:207c). a Dorsal view. b Lateral view. c Pleotelson. d ♀ × × Dorsal view of cephalon. e Ventral view of cephalon. f Dorsal view. g Lateral view. h Pleotelson. i Dorsal view of cephalon. j Ventral view of cephalon

times as long as greatest width, lateral margin weakly pereonite 1; pereonite lateral margins mostly posteri- convex, medial margin weakly convex. Exopod extend- orly ovate. Cephalon 0.32 as long as wide as measured ing beyond end of endopod, 7.2 times as long as greatest in dorsal view, trapezoid-shaped. Eyes oval, with distinct width, apically rounded, lateral margin weakly convex, margins. Pereonite 1 smooth, anterior border straight, medial margin weakly convex, terminating without setae. anterolateral angle narrowly rounded, in line with base of cephalon. Posterior margins of pereonites smooth Female paralectotype. [BMNH 1979.329:207c; Fig. 1.] and slightly curved laterally. Coxae 2–3 wide; with pos- Size 46.0 × 19.0. Body ovoid, 2.6 times as long as great- teroventral angles rounded; coxae 4–7 with rounded est width; dorsal surfaces smooth and polished in point and curved; not extending past posterior pere- appearance; widest at pereonite 5, most narrow at onite margin, coxa 7 extending past anterior pereonite Welicky and Smit Parasites Vectors (2019) 12:387 Page 6 of 34

margin, coxae 4–6 not extending past anterior pere- narrower; 3.5 times as long as wide. Pleopod 2 appendix onite margin. Pereonites 1–5 increasing in length and masculina distally narrowly rounded. width; pereonites 6–7 decreasing in length and width; pereonites 5 and 6 subequal, pereonites 1–4 narrower. Female. [New material SAMC-A091297; Figs. 3, 4.] Size Pleon with pleonite 1 wider than pleonites 2–5, visible 45.0 × 22.0. Body ovoid, 2.4 times as long as greatest in dorsal view; posterior margin of pleonites 1–4 poste- width; dorsal surfaces smooth and polished in appear- riorly concave; pleonite 5 posteriorly slightly concave to ance; widest at pereonite 5, most narrow at pereonite 1; straight or smooth, mostly concave. Pleonite 2 not over- pereonite lateral margins mostly posteriorly ovate. Ceph- lapped by pereonite 7; posterolateral angles of pleonite alon 0.42 as long as wide as measured in dorsal view, 2 narrowly rounded. Pleonite 1 similar in form to ple- trapezoid-shaped. Eyes oval, with distinct margins, one onites 2 and 3. Pleonites 3–5 similar in form to pleonite eye width 0.2 times width of cephalon, one eye length 0.4 2; pleonite 5 equal in width to pleonite 4 or free, not times length of cephalon. Pereonite 1 smooth, anterior overlapped by lateral margins of pleonite 4, pleonite 5 border straight, anterolateral angle narrowly rounded, with posterolateral angles narrowly rounded, posterior in line with base of cephalon. Posterior margins of pere- margin straight. Pleotelson 0.9 times as long as anterior onites smooth, slightly curved laterally. Coxae 2–3 wide, width, dorsal surface smooth, lateral margins convex, with posteroventral angles rounded; coxae 4–7 with posterior margin evenly rounded. rounded point and curved; coxae 4–6 not extending past Antennula approximately as stout as antenna; peduncle posterior pereonite margin; coxa 7 extending past ante- articles 1 and 2 distinct and articulated. rior pereonite margin; coxae 4–6 not extending past ante- Uropod more than half the length of pleotelson, pedun- rior pereonite margin. Pereonites 1–5 increasing in length cle 0.7 times as long as rami, peduncle lateral margin and width; pereonites 6–7 decreasing in length and width; without setae; rami extending beyond pleotelson, mar- pereonites 5 and 6 subequal, pereonites 1–4 narrower. ginal setae absent, apices broadly rounded. Endopod api- Pleon with pleonite 1 wider than pleonites 2–5, visible in cally rounded, 3.8 times as long as greatest width, lateral dorsal view; posterior margin of pleonites 1–4 posteriorly margin weakly convex, medial margin weakly convex. concave, pleonite 5 posteriorly slightly concave to straight Exopod extending beyond end of endopod, 7 times as or smooth, mostly concave. Pleonite 2 not overlapped by long as greatest width, apically rounded, lateral margin pereonite 7; posterolateral angles of pleonite 2 narrowly weakly convex, medial margin weakly convex, terminat- rounded. Pleonite 1 similar in form to pleonites 2 and ing without setae. 3. Pleonites 3–5 similar in form to pleonite 2; pleonite 5 equal width to pleonite 4 or free, not overlapped by lateral Male paralectotype. [BMNH 1979.329:207a; Fig. 2.] margins of pleonite 4, with posterolateral angles narrowly Size 35.0 × 10.0. Body similar to female but smaller and rounded, posterior margin straight. Pleotelson 0.8 times

b a d c

Fig. 2 Anilocra capensis (35.0 10.0, BMNH 1979.329:207a). a Dorsal view. b Lateral view. c Pleotelson. d Dorsal view of cephalon. e Ventral view ♂ × of cephalon Welicky and Smit Parasites Vectors (2019) 12:387 Page 7 of 34

a b

g h f

Fig. 3 Anilocra capensis (50.0 22.0, SAMC-A091297). a Dorsal view. b Lateral view. c Dorsal view of cephalon. d Ventral view of cephalon. e ♀ × Pereopod 1. f Pereopod 2. g Pereopods 3. h Pereopod 6. i Pleotelson Welicky and Smit Parasites Vectors (2019) 12:387 Page 8 of 34

j i k l

a e

h b

c n p d

m o

qr s tu Fig. 4 Anilocra capensis (45.0 22.0, SAMC-A091297). a–d Antenna. a Antenna. b Article 6 robust setae. c Article 7 robust setae. d Article 8 ♀ × simple setae. e–h Antennula. e Antennula. f Plumose setae article 2. g Robust setae article 3. h Robust setae article 6. i Maxillule. j Maxillule apex. k Mandible. l Article 3 mandibular palp. m Maxilla. n Maxilla apex. o Maxilliped. p Article 3 of maxilliped. q–u Pleopods 1–5, respectively as long as anterior width, dorsal surface smooth, lateral with 5 articles, extending to middle of eye; peduncle margins convex, posterior margin evenly rounded. article 3 0.8 times as long as wide, article 2 with plu- Antennula approximately as stout as antenna, com- mose setae, or articles 3 and 6 with robust setae. prised of 8 articles; peduncle articles 1 and 2 distinct, Antenna comprised of 8 articles, peduncle article 3 articulated; article 2 0.8 times as long as article 1; arti- 1.4 times as long as article 2, article 4 1.3 times as long cle 3 0.7 times as long as wide, 0.4 times as long as com- as wide, article 5 1.5 times as long as wide, 1.0 times bined lengths of articles 1 and 2; antennula fagellum as long as article 4; antenna fagellum with 3 articles, Welicky and Smit Parasites Vectors (2019) 12:387 Page 9 of 34

terminal article terminating in 1–5 short simple setae, Remarks extending to middle of pereonite 1. Mandibular molar Anilocra capensis has a proportionally large ovoid and process ending in an acute incisor, with 23 simple setae. vaulted body with distinctly fexed distolateral mar- Maxillula simple, with 4 terminal robust setae. Max- gins of the pereonite 7. Te pleotelson is wider than illa medial lobe partly fused to lateral lobe; 2 recurved long and broadly rounded and the distal-most article robust setae; and 2 large recurved robust setae. Maxil- of the antenna peduncle has one convex lateral mar- liped weakly segmented, with lamellar oostegite lobe or gin. Tis species has the greatest range in proportional second, smaller oostegite lobe on basal part of article, size of adult females and shows the most intrinsic vari- article 3 with 3 recurved robust setae. ation with regards to pleonite width and uropod length. Pereopod 1 basis 0.4 times as long as greatest width; Tis was evident in the syntypes as well as the new fresh ischium 1.9 times as long as basis; merus proximal mar- material examined. Te freshly fxed specimens can be gin without bulbous protrusion; carpus with straight grouped mainly into two categories: specimens where proximal margin; propodus 2.0 times as long as wide; pleonite 3 is most narrow and 5 is widest, and specimens dactylus stout, 2.4 times as long as basal width. Pereopod where pleonite 1 and 2 are widest. Within each of these 2 propodus 1.8 as long as wide; dactylus 1.3 as long as groups, proportional size and uropod length is variable, propodus. Pereopods gradually increasing in size towards but no general diferences in variability can be discerned posterior. Pereopod 6 basis 2.6 times as long as greatest between the two groups. Tere also appears to be col- width, ischium 0.4 times as long as basis, propodus 1.3 as our variation (dark brown to red-brown, or tan) in A. long as wide, dactylus 1.4 as long as propodus. Pereopod capensis, but all the other characters to this species are 7 basis 2.6 times as long as greatest width; ischium 0.6 observed in both colour types. Given that varying pleon as long as basis, without protrusions; merus 1.1 times as morphology is the only character that can create groups long as wide, 0.5 as long as ischium; carpus 1.3 times as within A. capensis, this is not sufcient to determine that long as wide, 1 as long as ischium, without bulbous pro- there are two species. trusion; propodus 2.4 times as long as wide, 0.7 as long as In the present study Anilocra capensis was found ischium; dactylus moderately slender, 1.1 times as long as attached to the skin of host fsh. It was situated below propodus, 2.9 times as long as basal width. the anterior portion of the dorsal fn, facing anteriorly, Pleopods without setae or lobes, exopod larger than and above and not blocking the operculum. Tis attach- endopod. Pleopod 1 exopod 1.3 times as long as wide, ment site was consistent among all the freshly collected lateral margin weakly convex, distally narrowly rounded, material. medial margin weakly oblique, medial margin weakly Anilocra capensis can be diferentiated from the other convex; endopod 1.2 times as long as wide, lateral margin species described below based on body, pereonite, pereo- weakly convex, distally narrowly rounded, medial mar- pod and/or antenna/antennule form. Te body shape of gin slightly convex, peduncle 4.5 times as wide as long, A. capensis is not twisted, whereas Anilocra jovanasi n. without retinaculae. Pleopods 2–5 similar to pleopod 1. sp. and Anilocra hadfeldae n. sp. have weakly twisted Pleopods 3–5 endopods proximal borders do not extend bodies. Compared to A. capensis, Anilocra paulsikkeli n. below exopod to peduncle. Peduncle lobes absent on ple- sp. is 0.86 times as wide at the widest point and has nota- opods 1–3, or present, increasing on pleopods 4–5. ble sub-parallel body margins, and Anilocra ianhudsoni Uropod more than half the length of pleotelson, pedun- n. sp. is strongly narrowed anteriorly. Anilocra capensis cle 0.5 times as long as rami, peduncle lateral margin can be distinguished from Anilocra bunkleywilliamsae without setae or without medial short acute robust seta; n. sp., as the latter is proportionally shorter, pereonite 1 rami extending beyond pleotelson, marginal setae absent, is longer, the antenna and antennule are narrower, and it apices broadly rounded. Endopod apically rounded, 3.5 has chromatophores. Anilocra capensis can be diferen- times as long as greatest width, lateral margin weakly tiated from Anilocra angeladaviesae n. sp., as A. capen- convex, medial margin weakly convex. Exopod extend- sis has a pronounced curvature on the lateral margin of ing beyond end of endopod, 6 times as long as greatest the propodus of pereopod 6 that A. angeladaviesae n. sp. width, apically rounded, lateral margin weakly convex, lacks. Tere is an elongated lateral margin of article 3 of medial margin weakly convex, terminating without setae. the antenna of A. angeladaviesae n. sp. that extends onto article 4, and this does not occur in A. capensis. Male. [New material SAMC-A091297; Figs. 5, 6.] Size We note that the 200-year-old female paralectotypes 28.0 × 8.0. Body similar to females but much smaller and were similar in shape and form to the fresh material, narrower; 3.1 times as long as wide. Pleopod 2 appen- but their poor and fragile condition, along with miss- dix masculina with parallel margins, 0.9 times as long as ing some articles made it possible to only include a few endopod, distally narrowly rounded. reliable drawings without further damage. Te male Welicky and Smit Parasites Vectors (2019) 12:387 Page 10 of 34

a b

f i

Fig. 5 Anilocra capensis (28.0 8.0, SAMC-A091297). a Dorsal view. b Lateral view. c Pleotelson. d Dorsal view of cephalon. e Ventral view of ♂ × cephalon. f Pereopod 1. g Pereopod 2. h Pereopod 6. i Pereopod 7 Welicky and Smit Parasites Vectors (2019) 12:387 Page 11 of 34

b a d c

h g

f e

k l m n o Fig. 6 Anilocra capensis (28.0 8.0, SAMC-A091297). a Antenna. b Antennula. c Mandible. d Article 3 mandibular palp. e Maxilla. f Maxilla apex. g ♂ × Maxilliped. h Article 3 of maxilliped. i Maxillule. j Maxillule apex. k–o Pleopods 1–5, respectively paralectotype examined was signifcantly larger and It is important to note that the type-locality for A. likely in a diferent and later developmental stage than capensis is Cape Town, South Africa, and the host is the freshly collected males. Tere was consistency in size a sparid. Species of Anilocra are regarded as typically and form of the fresh male material examined, but during host specifc to the family or genus level, and some our collections and those loaned from SAM, we found no even to the species level. Given this specifcity and male similar in size to Leach’s specimen. the general spatial distribution of A. capensis [31], it Welicky and Smit Parasites Vectors (2019) 12:387 Page 12 of 34

is unlikely that the records from the North Atlantic or Pleotelson 0.9 times as long as anterior width, dorsal other regions are of A. capensis. Tus, the reports of A. surface smooth, lateral margins convex, posterior margin capensis from the Canary Islands and from hosts repre- converging to weak caudomedial point. senting more than fve families of fsh are dubious [32, Antennula more stout than antenna; comprised of 7 33]. articles; peduncle articles 1 and 2 distinct, articulated; article 2 0.6 times as long as article 1; article 3 1 times Anilocra ianhudsoni n. sp. as long as wide, 0.5 times as long as combined lengths of articles 1 and 2; fagellum with 4 articles, extend- Type-host: Unknown. ing to middle of eye, terminal article with 1 plumose Type-locality: Nahoon River, East London, South seta. Antenna comprised of 9 articles; peduncle article 3 Africa. 1.4 times as long as article 2; article 4 1.5 times as long as wide, 1.1 times as long as article 3; article 5 1.5 times Type-material: Holotype: ♀ (18.0 × 8.0), paratypes: ♀ as long as wide, 1.2 times as long as article 4; fagellum (18.0 × 8.0, dissected), ♂ (11.5 × 3.0, dissected) (SAMC- A091293). Collected by personnel on the ‘SS Pieter with 4 articles, terminal article terminating in 1–5 short Faure’. simple setae, extending to middle of pereonite 1. Man- dibular molar process ending in an acute incisor, with Other-material: Transitional (11.5 × 3.0) (SAMC- A091293). 13 simple setae. Maxillula simple with 4 terminal robust ZooBank registration: Te Life Science Identifer setae. Maxilla medial lobe partly fused to lateral lobe; 2 (LSID) for Anilocra ianhudsoni n. sp. is urn:lsid:zoobank. recurved robust setae; and 2 large recurved robust setae. org:act:7AF506D2-5DD9-4B4A-8965-B1D224DE3650. Maxilliped weakly segmented, with lamellar oostegite Etymology: Tis species is named for the frst authorʼs lobe or second, smaller oostegite lobe on basal part of (RLW) nephew, Ian Hudson Catalano, as a gesture of article, palp article 2 with 0 simple setae, article 3 with 3 her appreciation to him and his mother, LM Catalano, recurved robust setae. for understanding the importance of her living abroad Pereopod 1 basis 1.7 times as long as greatest width; to conduct cymothoid-related research and to drive Ian’s ischium 0.7 times as long as basis; merus proximal mar- curiosity in the natural sciences. gin without bulbous protrusion; carpus with straight proximal margin; propodus 1.8 times as long as wide; Description dactylus moderately slender, 1.2 as long as propodus, 2.7 times as long as basal width. Pereopod 2 propodus 1.2 as Female. [SAMC-A091293; Figs. 7, 8.] Size 18.0 × 8.0. Body diamond-shaped or ovoid, 2.6 times as long as great- long as wide; dactylus 1.9 as long as propodus. Pereopods est width; dorsal surfaces smooth and polished in appear- gradually increasing in size towards posterior. Pereopod ance; widest at pereonite 5, most narrow at pereonite 1; 6 basis 2.3 times as long as greatest width, ischium 0.5 pereonite lateral margins posteriorly protruding. Cepha- times as long as basis, propodus 1.2 as long as wide, dac- lon 0.9 as long as wide as measured in dorsal view, sub- tylus 1.8 as long as propodus. Pereopod 7 basis 2.8 times triangular. Eyes oval, with distinct margins, one eye width as long as greatest width; ischium 0.6 as long as basis, 0.3 times width of cephalon, one eye length 0.4 times without protrusions; merus proximal margin without length of cephalon. Pereonite 1 smooth, anterior border bulbous protrusion, merus 1.5 times as long as wide, 0.6 straight, anterolateral angle narrowly-rounded, in line as long as ischium; carpus 1.7 times as long as wide, 0.5 with base of the cephalon. Posterior margins of pereonites as long as ischium, without bulbous protrusion; propo- smooth, slightly curved laterally. Coxae 2–3 wide, with dus 3.1 times as long as wide, 0.9 as long as ischium; dac- posteroventral angles rounded; coxae 4–7 with rounded tylus slender, 1.0 as long as propodus, 4.5 times as long as point and curved, extending past pereonite anterior mar- basal width. gin. Pereonites 1–5 increasing in length and width; pere- Pleopods without setae, exopod larger than endopod. onites 6–7 decreasing in length and width; pereonites 5 Pleopod 1 exopod 0.7 times as long as wide, lateral mar- and 6 subequal, pereonites 1–4 narrower, becoming more gin weakly convex, distally narrowly rounded, medial progressively rounded posteriorly. Pleon with pleonite 1 margin weakly oblique, medial margin weakly convex; wider than pleonites 2–4, visible in dorsal view; posterior endopod 0.5 times as long as wide, lateral margin weakly margin of pleonites 1–5 posteriorly concave, mostly con- convex, distally narrowly rounded, medial margin slightly cave. Pleonite 2 not overlapped by pereonite 7; postero- convex, without retinaculae. Pleopods 2–5 similar to ple- lateral angles of pleonite 2 narrowly-rounded. Pleonite 1 opod 1. Pleopods 3–5 endopods proximal borders do not similar in form to pleonites 2 and 3. Pleonites 3–5 similar extend below exopod to peduncle. Peduncle lobes absent. in form to pleonite 2; pleonite 5 free, not overlapped by Uropod longer than pleotelson, peduncle 0.5 times as lateral margins of pleonite 4, posterior margin straight. long as rami, peduncle lateral margin without setae; rami Welicky and Smit Parasites Vectors (2019) 12:387 Page 13 of 34

a g b

f d

Fig. 7 Anilocra ianhudsoni n. sp. (18.0 8.0, holotype SAMC-A091293). a Dorsal view. b Lateral view. c Dorsal view of cephalon. d Ventral view of ♀ × cephalon. e Pleotelson (18.0 8.0, paratype SAM A6296). f Pereopod 1. g Pereopod 2. h Pereopod 6. i Pereopod 7 × extending beyond pleotelson, marginal setae absent, api- beyond end of endopod, 6.0 times as long as greatest ces narrowly rounded. Endopod apically rounded, 3.0 width, apically rounded, lateral margin weakly convex, times as long as greatest width, lateral margin weakly medial margin weakly convex, terminating without setae. convex, medial margin weakly convex. Exopod extending Welicky and Smit Parasites Vectors (2019) 12:387 Page 14 of 34

a c i h g

f e

j l

n opq r Fig. 8 Anilocra ianhudsoni n. sp. (18.0 8.0, paratype SAMC-A091293). a Antennula. b Plumose setae of antennula terminal article. c Antenna. ♀ × d Forked setae of antenna article 4. e Antenna terminal article. f Maxiliped. g Article 3 of maxiliped. h Maxillule apex. i Maxillule. j Maxilla. k Maxilla apex. l Mandible. m Article 3 of mandible. n–r Pleopods 1–5, respectively Welicky and Smit Parasites Vectors (2019) 12:387 Page 15 of 34

straight. Coxae 2–3 narrow, with posteroventral angles Male. [SAMC-A091293; Fig. 9.] Size 11.5 × 3.0. Smaller than female. Body rectangular, 3.4 times as long as wide. rounded; coxae 4–7 with rounded point, not extending Pleopod 2 appendix masculina with parallel margins, dis- past posterior pereonite margin. Pereonites 1–5 increas- tally narrowly rounded. ing in length and width; pereonites 6–7 decreasing in length and width; pereonites 4–6 subequal, pereonites Remarks 1–3 subequal, increasing in similarity to pereonites 4–6. Anilocra ianhudsoni n. sp. is among the smaller of the Pleon with pleonite 1 wider than pleonites 2–5, visible in Anilocra species that occur in South Africa. Te body dorsal view; posterior margin of pleonites 1–4 posteriorly shape is unique as it has a diamond-triangulate shape, concave, pleonite 5 posteriorly slightly concave to straight, such that pereonites 1–3 narrow strongly. Unlike the mostly concave. Pleonite 2 not overlapped by pereonite other African species, A. ianhudsoni n. sp. has no fexed 7; posterolateral angles of pleonite 2 narrowly rounded. pereonite posterolateral margins. Te antennula has a Pleonite 1 similar in form to pleonites 2 and 3. Pleonites plumose seta on the terminal article and a maxilla with 3–5 similar in form to pleonite 2; pleonite 5 free, not over- simple setae or 3 recurved robust setae. Anilocra ianhud- lapped by lateral margins of pleonite 4, posterior margin soni n. sp. also has more slender pereopods and a fat- straight. Pleotelson 1 times as long as anterior width. Dor- ter and broader antenna compared to the other species sal surface smooth. Pleotelson lateral margins convex, described herein. posterior margin converging to weak caudomedial point. Anilocra ianhudsoni n. sp. has a produced curvature on Antennula more stout than antenna; peduncle articles the lateral margin of the propodus of pereopod 6, which 1 and 2 distinct and articulated; article 2 0.8 times as long is similar to A. capensis and A. bunkleywilliamsae n. sp. as article 1; article 3 1.0 times as long as wide, 0.4 times Compared to Anilocra ianhudsoni n. sp., A. capensis and as long as combined lengths of articles 1 and 2; fagellum A. bunkleywilliamsae n. sp. both have broader, more with 4 articles, extending to middle of eye, articles 3 and ovoid pereonites, and a pleotelson that is closer in width 6–8 with robust setae. Antenna comprised of 9 articles; to total body width. peduncle article 3 1 times as long as article 2; article 4 0.7 times as long as wide, 0.9 times as long as article 3; Anilocra bunkleywilliamsae n. sp. article 5 1.6 times as long as wide, 1.5 times as long as article 4; fagellum with 4 articles, terminal article termi- Type-host: Rhabdosargus holubi Steindachner, 1881. nating in 6–10 short simple setae, extending to middle Type-locality: Mawalana Estuary, South Africa. of pereonite 1. Mandibular molar process ending in an acute incisor, with 17 simple setae. Maxillula simple, with Type-material: Holotype ♀ (24.0 × 10.0) paratype ♀ 4 terminal robust setae. Maxilla medial lobe partly fused (27.0 × 12.0, dissected) (SAMC-A091294). Collected by T. Mqolombia, on 17 February 1998. to lateral lobe; lateral lobe with 0 simple setae, 2 recurved robust setae; medial lobe with 0 simple setae, and 2 large Other material: Transitional (18.0 × 5.0). ZooBank registration: Te Life Science Identifer (LSID) recurved robust setae. Maxilliped weakly segmented, for Anilocra bunkleywilliamsae n. sp. is urn:lsid:zoobank. with lamellar oostegite lobe or second, smaller oostegite org:act:9DF125DA-624A-4712-BA07-D45CF4BC7C8F. lobe on basal part of article, palp article 2 with 0 simple Etymology: Tis species is named in honour of Dr Lucy setae, article 3 with 4 recurved robust setae. Oostegites Bunkley-Williams, a world expert on Anilocra who margin covered in numerous plumose setae. described nearly all of the known species of Anilocra Pereopod 1 basis 1.7 times as long as greatest width; from the Caribbean. ischium 0.7 times as long as basis; merus proximal margin without bulbous protrusion; carpus with straight Description proximal margin; propodus 1.2 times as long as wide; dactylus moderately slender, 1.2 as long as propodus, 2.6 Female. [SAMC-A091294; Figs. 10, 11.] Size 27.0 × 12.0. Body 2.2 times as long as greatest width with dorsal sur- times as long as basal width. Pereopod 2 propodus 1.9 as faces smooth and polished in appearance. Body widest at long as wide; dactylus 1.1 as long as propodus. Pereopods pereonite 5, most narrow at pereonite 1, and pereonite gradually increasing in size towards posterior. Pereopod lateral margins posteriorly protruding. Cephalon 0.8 6 basis 2.5 times as long as greatest width, ischium 0.4 as long as wide as measured in dorsal view, trapezoid- times as long as basis, propodus 1.3 as long as wide, dac- shaped. Eyes oval, with distinct margins, one eye width 0.2 tylus 1.4 as long as propodus. Pereopod 7 basis 2.8 times times width of cephalon, one eye length 0.4 times length as long as greatest width; ischium 0.5 as long as basis, of cephalon. Pereonite 1 in line with base of cephalon, without protrusions; merus proximal margin without smooth, anterior border straight, anterolateral angle nar- bulbous protrusion, merus 1.6 times as long as wide, 0.6 rowly rounded. Posterior margins of pereonites smooth, as long as ischium; carpus 2.0 times as long as wide, 0.6 Welicky and Smit Parasites Vectors (2019) 12:387 Page 16 of 34

b e a d f

k h

Fig. 9 Anilocra ianhudsoni n. sp. (11.5 3.0, SAMC-A091293). a Dorsal view. b Lateral view. c Pleotelson. d Antenna. e Antennule. f Dorsal view of ♂ × cephalon. g Ventral view of cephalon. h Pereopod 1. i Pereopod 2. j Pereopod 6. k Pereopod 7 Welicky and Smit Parasites Vectors (2019) 12:387 Page 17 of 34

b a c

i f

Fig. 10 Anilocra bunkleywilliamsae n. sp. (24.0 10.0, holotype, SAMC-A091294). a Dorsal view. b Lateral view. c Dorsal view of cephalon. d Ventral ♀ × view of cephalon. e Pleotelson (27.0 12.0, paratype SAMC-A091294). f Pereopod 7. g Pereopod 6. h Pereopod 2. i Pereopod 1 × Welicky and Smit Parasites Vectors (2019) 12:387 Page 18 of 34

f n p

c o k m d

l e

t r

q s

x y u vw Fig. 11 Anilocra bunkleywilliamsae n. sp. (27.0 12.0, paratype SAMC-A091294). a–e Antennula. a Antennula. b Setae of article 3. c–e Setae of ♀ × articles 6–8, respectively. f Antenna. g–l Setae of antenna articles 4–9, respectively. m Maxilliped. n Article 3 of maxiliped. o Maxillule. p Maxillule apex. q Maxilla. r Maxilla apex. s Mandible. t Article 3 mandibular palp. u–y Pleopods 1–5, respectively Welicky and Smit Parasites Vectors (2019) 12:387 Page 19 of 34

as long as ischium, without bulbous protrusion; propo- Type-material: Holotype ♀ (32.0 × 8.5), paratypes: ♀ dus 2.8 times as long as wide, 1 as long as ischium; dac- (33.0 × 9.0, dissected), ♂ (16.0 × 3.0, dissected; 18.0 × 4.0) tylus slender, 1 as long as propodus, 3.6 times as long as (SAMC-A091295). Unknown collector. basal width. Dense chromatophores present on 1–7. ZooBank registration: Te Life Science Identifer Exopod larger than endopod. Pleopod 1 exopod 1.4 times (LSID) for Anilocra paulsikkeli n. sp. is urn:lsid:zoobank. as long as wide, lateral margin weakly convex, distally nar- org:act:AFEA08B1-D27B-49F6-A30E-B4E142EFA5D9. rowly rounded, medial margin weakly oblique, medial Etymology: Tis species is named in honour of Dr Paul C. margin weakly convex; endopod 1.6 times as long as wide, Sikkel, the PhD supervisor of the frst author (RLW) and lateral margin weakly convex, distally narrowly rounded, research collaborator of the second author (NJS). Dr Sik- medial margin slightly convex, peduncle 2.9 times as wide kel’s devotion to furthering our knowledge on host-para- as long, without retinaculae. Pleopods 2–5 similar to ple- site interactions, strengthening the relationships between opod 1. Pleopods 3–5 endopods proximal borders do not scientists and non-scientists, and rigorously mentoring extend below exopod to peduncle. Peduncle lobes absent. future fsh-parasite ecologists is hereby acknowledged. Uropod more than half the length of pleotelson, peduncle 0.5 times as long as rami, peduncle lateral margin Description without setae; rami extending beyond pleotelson, mar- Female. [SAMC-A091295; Figs. 12–13.] Size 33.0 × 9.0. ginal setae absent, apices narrowly rounded. Endopod Body rectangular or elongate, 3.7 times as long as great- apically rounded, 5 times as long as greatest width, lateral est width, dorsal surfaces smooth and polished in appear- margin weakly convex, medial margin weakly convex, ance, widest at pereonite 5, most narrow at pereonite terminating with 0 setae. Exopod extending beyond end 1, pereonite lateral margins subparallel. Cephalon 0.73 of endopod, 9.7 times as long as greatest width, apically as long as wide as measured in dorsal view, trapezoid- rounded, lateral margin weakly convex, medial margin shaped. Eyes oval, with distinct margins, one eye width weakly convex, terminating without setae. 0.2 times width of cephalon; one eye length 0.45 times length of cephalon. Pereonite 1 in line with base of Remarks cephalon, smooth, anterior border straight, anterolateral Anilocra bunkleywilliamsae n. sp. is most readily distin- angle narrowly rounded. Posterior margins of pereonites guished from all other African species as it has the most smooth and straight. Coxae 2–3 wide; with posteroven- anteriorly rounded and arched rostrum, and more setae tral angles rounded; 4–7 with rounded point and curved; on the antennula and antenna of the species described not extending past posterior pereonite margin. Pere- herein. It also has large and dense chromatophores on its onites 1–4 increasing in length and width; pereonites pereopods. Te chromatophores are denser on the left 6–7 decreasing in length and width; pereonites 5 and side when in dorsal view. 6 subequal. Pleon with pleonite 1 largely concealed by Compared to A. capensis, the body of A. bunkleywil- pereonite 7, visible in dorsal view; pleonites posterior liamsae n. sp. is more ovoid, and pereopod 2 has robust margin 1–4 posteriorly concave, pleonite 5 posteriorly setae present. Compared to A. ianhudsoni n. sp., A. slightly concave to straight, mostly concave. Pleonite 2 bunkleywilliamsae n. sp. is generally smaller, its pleo- not overlapped by pereonite 7; posterolateral angles of telson converges more and is rounder, and its dactyls pleonite 2 expanded, posteriorly produced or rounded. are shorter. Te ovate and non-twisted body shape of A. Pleonite 1 difer in form to pleonite 2 and 3. Pleonite 5 bunkleywilliamsae n. sp. makes it distinguishable from free, not overlapped by lateral margins of pleonite 4, with A. paulsikkeli n. sp. which is long and narrow, and A. posterolateral angles narrowly rounded, posterior margin jovanasi n. sp. and A. hadfeldae n. sp. which are weakly straight. Pleotelson 1.31 times as long as anterior width. twisted. Anilocra bunkleywilliamsae n. sp.is most similar Dorsal surface smooth. Pleotelson lateral margins con- in pereonite shape to A. angeladaviesae n. sp., but these vex, posterior margin converging to weak caudomedial species can be diferentiated from each other as A. ange- point. ladaviesae n. sp. has no produced curvature on the lat- Antennula approximately as stout as antenna, com- eral margin of the propodus of pereopod 6, its pereopods prised of 9 articles; peduncle articles 1 and 2 distinct and are more slender, and the general shape and form of the articulated; article 2 1.3 times as long as article 1; article antenna and antennula are dissimilar. 3 0.9 times as long as wide, 0.6 times as long as combined lengths of articles 1 and 2; fagellum with 5 articles, ter- Anilocra paulsikkeli n. sp. minal article with robust and simple setae, extending to posterior margin of eye. Antenna comprised of 9 articles; Type-host: Unknown. peduncle article 3 1.0 times as long as article 2; article 4 Type-locality: Delagoa Bay, Mozambique. 1.15 times as long as wide, 1.5 times as long as article 3; Welicky and Smit Parasites Vectors (2019) 12:387 Page 20 of 34

b a c

i f g

Fig. 12 Anilocra paulsikkeli n. sp. (a–e 32.0 8.5, holotype, SAMC-A091295; f–i 33.0 9.0, paratype, SAMC-A091295). a Dorsal view. b Lateral view. ♀ × × c Dorsal view of cephalon. d Ventral view of cephalon. e Pleotelson. f Pereopod 1. g Pereopod 2. h Pereopod 6. i Pereopod 7 Welicky and Smit Parasites Vectors (2019) 12:387 Page 21 of 34

i c

g e

b h f

m o j k Fig. 13 Anilocra paulsikkeli n. sp. (33.0 9.0, paratype, SAMC-A091295). a–b Antenna. a Antenna. b Setae of article 8. c–e Antennula. c Antennula. ♀ × d Setae of article 3. e Setae of terminal article. f Mandible. g Article 3 mandibular palp. h Maxilliped. i Article 3 of maxilliped. j Pleopod 1. k Maxilla. l Maxilla apex. m Maxillule. n Maxillule apex. o Pleopod 2 article 5 1.4 times as long as wide, 1.2 times as long as of article, article 3 with 3 recurved robust setae. Ooste- article 4; fagellum with 4 articles, terminal article ter- gites margin covered in numerous plumose setae. minating in no setae, extending to middle of pereonite Pereopod 1 basis 2.5 times as long as greatest width; 1. Mandibular molar process ending in an acute incisor, ischium 0.5 times as long as basis; merus proximal mar- with 11 simple setae. Maxillula simple, with 2 terminal gin with slight bulbous protrusion; carpus with straight robust setae. Maxilla medial lobe partly fused to lateral proximal margin; propodus 2.1 times as long as wide; lobe; 2 recurved robust setae and 2 large recurved robust dactylus stout, 1.2 as long as propodus, 3 times as long setae. Maxilliped weakly segmented, with lamellar oost- as basal width. Pereopod 2 propodus 2.5 as long as wide; egite lobe or second, smaller oostegite lobe on basal part dactylus 1.1 as long as propodus. Pereopods gradually increasing in size towards posterior. Pereopod 6 basis Welicky and Smit Parasites Vectors (2019) 12:387 Page 22 of 34

2.7 times as long as greatest width, ischium 0.5 times as 1987’s drawings of the lectotype. In Aneesh et al. [34], long as basis, propodus 2.6 as long as wide, dactylus 1 as Fig. 1d, e appear to not resemble A. leptosoma, and are long as propodus. Pereopod 7 basis 2.4 times as long as more similar to A. capensis, with respect to body shape greatest width; ischium 0.7 as long as basis, without pro- and form in dorsal view. Aneesh et al. [34] likely did trusions; merus proximal margin without bulbous pro- not identify A. leptosoma. To identify the species these trusion, merus 1.7 times as long as wide, 0.6 as long as authors report on, specimens should be further examined ischium; carpus 1.5 times as long as wide, 0.44 as long as morphologically, and molecular data should be provided. ischium, without bulbous protrusion; propodus 4.3 times Anilocra paulsikkeli n. sp. is also similar to Anilocra as long as wide, 1.1 as long as ischium; dactylus slender, caudata Bovallius, 1887 but can be distinguished from 0.9 as long as propodus, 4.4 times as long as basal width. A. caudata, by the wider pleotelson, pleonite 1 not being Pleopods without setae, exopod larger than endopod. covered by pereonite 7 and the lateral margins of pleonite Pleopod 1 exopod 2.1 times as long as wide, lateral mar- 5 are more acute. gin weakly convex, distally narrowly rounded, medial margin weakly oblique, medial margin weakly convex; Compared to the other species described herein, A. endopod 2.2 times as long as wide, lateral margin weakly paulsikkeli n. sp. and A. jovanasi n. sp. are most simi- convex, distally narrowly rounded, medial margin slightly lar. Both species have nodules that occur midway on the convex, peduncle 2.0 times as wide as long, without reti- lateral margins of the dactyls of pereopods 1 and 2, but naculae. Pleopods 3–5 endopods proximal borders do not the nodules are larger on A. paulsikkeli n. sp. Anilocra extend below exopod to peduncle. Peduncle lobes absent. jovanasi n. sp. also has antennula peduncle article 3 more Uropod more than half the length of pleotelson, pedun- strongly produced than that of A. paulsikkel n. sp. cle 0.7 times as long as rami, lateral margin without setae; rami extending to pleotelson apex, marginal setae absent, Anilocra jovanasi n. sp. apices broadly rounded. Endopod apically rounded, 3.3 times as long as greatest width, lateral margin weakly Type-host: Unknown. convex, medial margin weakly convex, terminating with 0 Type-locality: Delagoa Bay, Mozambique. setae. Exopod not extending to end of endopod, 6.7 times Type-material: Holotype ♀ (20.0 × 7.0, dissected) as long as greatest width, apically rounded, lateral margin (SAMC-A091296). Unknown collector. weakly convex, medial margin weakly convex, terminat- ZooBank registration: Te Life Science Identifer ing without setae. (LSID) for Anilocra jovanasi n. sp. is urn:lsid:zoobank. org:act:DA044D6D-CA90-4E20-9D76-1D6FD636569C. Male. [SAMC-A091295; Fig. 14.] Size 16.0 × 3.0. Simi- Etymology: Tis species is named for Professor Jo G. van lar to female but much smaller. Body rectangular, weakly As (1949–2018), the late aquatic parasitologist and PhD twisted, 4.9 times as long as wide. Pleopod 2 appendix advisor of the second author (NJS). Professor van As masculina with parallel margins, 0.8 times as long as revived the feld of aquatic parasitology in South Africa endopod, distally narrowly rounded. in the 1970s and all current active aquatic parasitologists in South Africa are either his former students, or a stu- Remarks dent of one of his former students. His contribution to Anilocra paulsikkeli n. sp. has a distinctive elongate, nar- the feld of aquatic parasitology in South Africa is hereby row body, with sub-parallel lateral margins. Pleonite 1 is acknowledged. partly covered by pereonite 7. Te lateral margins of the pleotelson are sub-parallel and from the last third of the Description pleotelson there is a strong convergence to a rounded Female. [SAMC-A091296; Figs. 15, 16.] Size 20.0 × 7.0. medial point. Te eyes of A. paulsikkeli n. sp. are nearly Body ovoid and weakly twisted, 2.9 times as long as one-quarter the width of the cephalon. greatest width, dorsal surfaces smooth and polished in Anilocra paulsikkeli n. sp. is similar to A. leptosoma, but appearance, most narrow at pereonite 1, pereonite lateral there are multiple diferences. Anilocra paulsikkeli n. sp. margins mostly posteriorly ovate. Cephalon 0.67 as long is larger (3.7 times as long as wide whereas A. leptosoma as wide as measured in dorsal view, trapezoid-shaped. is 3.4 times as long as wide), the frst antenna article 3 Eyes oval, with distinct margins, one eye width 0.2 times is not produced, pleonite 1 is more concealed, and the width of cephalon; one eye length 0.4 times length of posterior margin of the pleonites are straighter. Aneesh cephalon. Pereonite 1 in line with base of cephalon, et al. [34] recently resdescribed A. leptosoma but did smooth, anterior border straight, anterolateral angle nar- not examine the A. leptosoma lectotype material, and rowly rounded. Posterior margins of pereonites smooth the drawings of Aneesh are in disagreement with Bruce and straight, 5 extended posteriorly and overlay lateral Welicky and Smit Parasites Vectors (2019) 12:387 Page 23 of 34

a b e f d

l c

k h i

Fig. 14 Anilocra paulsikkeli n. sp. (16.0 3.0, SAMC-A091295). a Dorsal view. b Lateral view. c Pleotelson. d Antenna, damaged. e Antennula. f ♂ × Dorsal view of cephalon. g Ventral view of cephalon. h Pleopod 1. i Pleopod 2. j Pereopod 1. k Pereopod 2. l Pereopod 6. m Pereopod 7 Welicky and Smit Parasites Vectors (2019) 12:387 Page 24 of 34

a b

e h

g i

Fig. 15 Anilocra jovanasi n. sp. (20.0 7.0, holotype, SAMC-A091296). a Dorsal view. b Lateral view. c Dorsal view of cephalon. d Ventral view of ♀ × cephalon. e Pleotelson. f Pereopod 1. g Pereopod 2. h Pereopod 6. i Pereopod 7 Welicky and Smit Parasites Vectors (2019) 12:387 Page 25 of 34

e l p a

j f n g h

b c

d m i k o

q r s t u Fig. 16 Anilocra jovanasi n. sp. (20.0 7.0, holotype, SAMC-A091296). a–d Antenna. a Antenna. b–d Setae of articles 8–10, respectively. e–h ♀ × Antennula. e Antennula. f–h Setae of articles 6–8, respectively. i Maxilla. j Maxilla apex. k Maxillule. l Maxillule apex. m Maxilliped. n Article 3 of maxilliped. o Mandible. p Mandibular palp. q–u Pleopods 1–5, respectively

margins of pleotelson. Coxae 2–3 wide; with posteroven- 5; pleonite 5 free, not overlapped by lateral margins of tral angles rounded; coxae 4–7 with rounded point and pleonite 4, posterolateral margins extend onto anterior curved; not extending past posterior pereonite margin. portion of pleotelson, posterior margin straight. Pleo- Pereonites 1–5 increasing in length and width; pereonites telson 1.3 times as long as anterior width, dorsal surface 6–7 decreasing in length and width; pereonites 5 and 6 smooth, lateral margins weakly convex, posterior margin subequal, pereonites 1–4 narrower. Pleon with pleonite 1 converging to weak caudomedial point. wider than 2–4, visible in dorsal view; posterior margin Antennula approximately as stout as antenna; com- of pleonites 1–4 posteriorly concave, pleonite 5 posteri- prised of 8 articles; peduncle articles 1 and 2 distinct and orly slightly concave to straight, mostly concave. Pleonite articulated; article 2 1.1 times as long as article 1; article 2 not overlapped by pereonite 7; posterolateral angles of 3 0.6 times as long as wide, 0.4 times as long as combined pleonite 2 narrowly rounded. Pleonite 1 similar in form lengths of articles 1 and 2; fagellum with 5 articles, arti- to pleonite 2. Pleonites 3–5 difer in form from pleonite cles 6–8 with robust setae, extending to middle of eye. Welicky and Smit Parasites Vectors (2019) 12:387 Page 26 of 34

Antenna comprised of 10 articles; peduncle article 3 1.7 third article; pereopods 6 and 7 possess few robust setae. times as long as article 2; article 4 1.4 times as long as Anilocra jovanasi n. sp. is the only species described in wide, 1.4 times as long as article 3; article 5 2.6 times as southern Africa with the posterolateral margins of the long as wide, 1.5 times as long as article 4; fagellum with ffth pleonite extending onto the dorsal portion of the 5 articles, articles 8–9 with setae, terminal article termi- pleotelson. nating in 1–5 short simple setae, extending to middle of Anilocra jovanasi n. sp. is most similar in body shape pereonite 1. Mandibular molar process present, with 20 to A. leptosoma, and A. paulsikkeli n. sp. However, the simple setae. Maxilla medial lobe partly fused to lateral body of A. leptosoma is not twisted and the pleotelson lobe; 2 recurved robust setae; and 2 large recurved robust converges strongly to a distinct point, whereas in A. setae. Maxilliped weakly segmented, with lamellar oost- jovanasi n. sp., the body is straight and the pleotelson egite lobe, palp article 2 with 0 simple setae, article 3 with converges broadly to a very weak point. Te pleotel- 3 recurved robust setae. Oostegites margin covered in sons of A. paulsikkeli n. sp. and A. jovanasi n. sp. are numerous plumose setae. similar in shape until the last third, at which point the Pereopod 1 basis 2.7 times as long as greatest width; lateral margins of the pleotelson of A. jovanasi n. sp. ischium 0.6 times as long as basis; merus proximal mar- converge broadly to a very weak point, and those of A. gin without bulbous protrusion; carpus with straight paulsikkeli n. sp. converge strongly to a broad medial proximal margin; propodus 2 times as long as wide; dac- point. Moreover, pleonite 1 of A. jovanasi n. sp. is sub- tylus slender, 1.2 as long as propodus, 3.2 times as long stantially more visible than that of A. paulsikkeli n. sp. as basal width. Pereopod 2 propodus 2.15 as long as Whereas A. paulsikkeli n. sp. has distinct nodules on wide; dactylus 1 as long as propodus. Pereopods gradu- the dactyls of pereopods 1 and 2, this is present only ally increasing in size towards posterior. Pereopod 6 basis on pereopod 1 of A. jovanasi n. sp. Te propodus of 2.91 times as long as greatest width, ischium 0.5 times as pereopod 2 is without a curved produced curvature on long as basis, propodus 2.6 as long as wide, dactylus 1.1 the lateral margin, separating it from A. capensis and as long as propodus. Pereopod 7 basis 3.1 times as long A. bunkleywilliamsae n. sp. Anilocra ianhudsoni n. sp. as greatest width; ischium 0.6 as long as basis, without has a distinctly diferent body form and its pereopods protrusions; merus proximal margin without bulbous 6 and 7 are more dense in robust setae. Anilocra ange- protrusion, merus 2.0 times as long as wide, 0.7 as long as ladaviesae n. sp. is more robustly ovoid, with a broader ischium; carpus 1.9 times as long as wide, 0.6 as long as pleotelson than A. jovanasi n. sp. Anilocra hadfeldae ischium, without bulbous protrusion; propodus 3.2 times n. sp. also has a weakly twisted body, but compared to as long as wide, 0.9 as long as ischium; dactylus slender, A. jovanasi n. sp., it has a more produced pointed lat- 0.86 as long as propodus, 5.0 times as long as basal width. eral margin of pleonite 7 and a medial indent on the Pleopods without setae, exopod larger than endopod. pleotelson. Pleopod 1 exopod 1.6 times as long as wide, lateral margin weakly convex, distally narrowly rounded, medial Anilocra angeladaviesae n. sp. margin weakly oblique, medial margin weakly convex; endopod 1.6 times as long as wide, lateral margin weakly Syn. Anilocra capensis Monod, 1924 convex, distally narrowly rounded, medial margin slightly Type-host: Uncertain (see Remarks). convex, peduncle 2.4 times as wide as long, without reti- Type-locality: Cape Blanc, Morocco. naculae. Pleopods 2–5 similar to pleopod 1. Pleopods Type-material: Holotype ♀ (BMNH1924.5.30.6) 3–5 endopods proximal borders do not extend below (37.0 × 18.0), paratypes (BMNH1924.5.30.7–10): ♀ exopod to peduncle. Peduncle lobes present on 5. (38.0 × 17.0; 37.0 × 15.0; 35.0 × 13.0), ♂ (24.0 × 6.0; Uropod more than half the length of pleotelson, pedun- 18.0 × 5.0). Tere are 6 accession numbers but 7 speci- cle lateral margin without setae; rami not extending mens in this lot. Collected by M. Teo Monod, 1923. beyond pleotelson, marginal setae absent, apices nar- ZooBank registration: Te Life Science Identifer (LSID) rowly rounded. Endopod apically rounded, lateral margin for Anilocra angeladaviesae n. sp. is urn:lsid:zoobank. weakly convex, medial margin weakly convex. Apically org:act:3381D22E-67B0-4847-A559-0899B20E9F7D. rounded, lateral margin weakly convex, medial margin Etymology: Tis species is named after Professor Angela weakly convex. Josephine Davies (1947–2013), PhD promotor and postdoctoral advisor to the second author (NJS), to commem- Remarks orate her contribution to the knowledge of crustacean Te body Anilocra jovanasi n. sp. is weakly twisted and parasites of fshes, as well as her singular dedication to- narrowly ovoid. In dorsal view, the antennula appears and enthusiasm for sharing this knowledge with all those geniculate due to the anterodistal lateral angle of the she mentored. Welicky and Smit Parasites Vectors (2019) 12:387 Page 27 of 34

Description protrusion, merus 1.3 times as long as wide, 0.6 as long as Female. [BMNH1924.5.30.6; Fig. 17.] Size 37 × 18. Body ischium; carpus 1.4 times as long as wide, 0.5 as long as ovoid, 2.1 times as long as greatest width, dorsal surfaces ischium, without bulbous protrusion; propodus 2.1 times smooth and polished in appearance, widest at pereonite as long as wide, 0.7 as long as ischium; dactylus slender, 1 5, most narrow at pereonite 1, pereonite lateral margins as long as propodus, 2.9 times as long as basal width. mostly posteriorly ovate. Cephalon 0.7 as long as wide Pleopods without setae, exopod larger than endopod. as measured in dorsal view, trapezoid-shaped. Eyes oval, Uropod more than half the length of pleotelson, pedun- with distinct margins, one eye width 0.7 times width cle lateral margin without setae; rami not extending beyond of cephalon; one eye length 0.3 times length of cepha- pleotelson, marginal setae absent, apices narrowly rounded. lon. Pereonite 1 in line with base of cephalon, smooth, Endopod apically rounded, 3.7 times as long as greatest anterior border straight, anterolateral angle narrowly width, lateral margin weakly convex, medial margin weakly rounded. Posterior margins of pereonites smooth and convex, terminating with 0 setae. Exopod extending beyond straight. Coxae 2–3 wide; with posteroventral angles end of endopod, 6.0 times as long as greatest width, api- rounded; coxae 4–7 with rounded point and curved; cally rounded, lateral margin weakly convex, medial margin not extending past posterior pereonite margin. Pere- weakly convex, terminating without setae. onites 1–5 increasing in length and width; pereonites 6–7 decreasing in length and width; pereonites 1–5 sub- Male. [BMNH1924.5.30.7–10; Fig. 18.] Size 30 × 18. Sim- equal, pereonites 6–7 subequal. Pleon with pleonite 1 ilar to female but much smaller. Body ovoid to rectangu- wider than pleonites 2–4, visible in dorsal view; posterior lar, weakly twisted, 3.2 times as long as wide. Pleopod 2 margin of pleonites 1–4 posteriorly concave, pleonite 5 appendix masculina with parallel margins, distally nar- posteriorly slightly concave to straight, mostly concave. rowly rounded. Pleonite 2 not overlapped by pereonite 7; posterolateral angles of pleonite 2 narrowly rounded. Pleonite 1 simi- Remarks lar in form to pleonite 2. Pleonites 3–5 similar in form Anilocra angeladaviesae n. sp. can best be identifed by to pleonite 2; pleonite 5 free, not overlapped by lateral the distinct elongate lateral margin on the third article margins of pleonite 4, posterior margin straight. Pleo- of the antenna peduncle that appears to partially cover telson 0.8 times as long as anterior width, dorsal surface the lateral margin of the fourth article; the antennule is smooth, lateral margins weakly convex, posterior margin nearly half the length of the antenna; the body is ovoid evenly rounded. and pereonites 1–3 narrow weakly towards the cephalon. Antennula more stout than antenna, comprised of 7 It is likely that the specimens examined here are those articles; peduncle articles 1 and 2 distinct and articulated; mentioned by Monod as A. capensis from Morocco [35] article 2 1.2 times as long as article 1; article 3 1.1 times because the material examined from NHM was collected as long as wide, 0.4 times as long as combined lengths of by Monod in 1923, and is clearly labelled as A. capen- articles 1 and 2; fagellum with 4 articles, extending to sis from Morocco. Monod [35] referenced the genus middle of eye. Antenna comprised of 9 articles. Pedun- names, Sama and Dentex, and the species, Morone punc- cle article 3 2.17 times as long as article 2; article 4 1.5 tata (current accepted name is Dicentrarchus puncta- times as long as wide, 0.9 times as long as article 3; article tus Bloch, 1792). 5 2.0 times as long as wide, 1.3 times as long as article 4. Compared to Schioedte & Meinert’s [7] drawings of Antenna fagellum with 4 articles, extending to middle of A. physodes, the pereonite margins and pleotelson of A. pereonite 1. angeladaviesae n. sp. are much straighter and less acute, Pereopod 1 basis 2.8 times as long as greatest width; respectively. Compared to A. capensis, A. angeladaviesae ischium 1.9 times as long as basis; merus proximal mar- n. sp. has a narrower pleonite 1 in some specimens. Ani- gin without bulbous protrusion; carpus with straight locra angeladaviesae n. sp. lacks the curved lateral mar- proximal margin; propodus 1.8 times as long as wide; gin of the pereopod 6 propodus, which is present on both dactylus stout, 1.1 as long as propodus, 2.3 times as long A. capensis and A. bunkleywilliamsae n. sp. Anilocra as basal width. Pereopod 2 propodus 2.3 as long as wide; bunkleywilliamsae n. sp. also has large dense chromato- dactylus 0.9 as long as propodus. Pereopods gradually phores that are not present on A. angeladaviesae n. sp. increasing in size towards posterior. Pereopod 6 basis Anilocra paulsikkeli n. sp. and A. jovanasi n. sp. are both 3 times as long as greatest width, ischium 0.6 times as more elongate and have nodules on pereopods 1 and 2 long as basis, propodus 1.2 as long as wide, dactylus 2 compared to A. angeladaviesae n. sp. Anilocra angela- as long as propodus. Pereopod 7 basis 2.0 times as long daviesae n. sp. has a more rounded rostrum and posterior as greatest width; ischium 0.7 as long as basis, without margin of pereonite 7 and more ovoid body compared to protrusions; merus proximal margin without bulbous A. hadfeldae n. sp. Welicky and Smit Parasites Vectors (2019) 12:387 Page 28 of 34

a b

f e

h j

Fig. 17 Anilocra angeldaviesae n. sp. (37.0 18.0, holotype, BMNH1924.5.30.6). a Dorsal view. b Lateral view. c Dorsal view of cephalon. d Ventral ♀ × view of cephalon. e Antenna. f Antennula. g Pleotelson. h Pereopod 1. i Pereopod 2. j Pereopod 6. k Pereopod 7

Anilocra hadfeldae n. sp. Type-material: Holotype ♀ (42.0 × 16.0, BMNH 1952.9.9.29–30); paratype ♂ (30.0 × 9.0, BMNH Type-host: Unknown. 1952.9.9.29–30). Collected by M. Routh, 1952. Type-locality: Cape Blanco, Gambia. Welicky and Smit Parasites Vectors (2019) 12:387 Page 29 of 34

ab c e

Fig. 18 Anilocra angeldaviesae n. sp. (30.0 9.0, paratype, BMNH1924.5.30.7). a Dorsal view. b Lateral view. c Dorsal view of cephalon. d Ventral ♂ × view of cephalon. e Pleotelson

ZooBank registration: Te Life Science Identifer in length and width; pereonites 6–7 decreasing in length (LSID) for Anilocra hadfeldae n. sp. is urn:lsid:zoobank. and width; pereonites 5 and 6 subequal, pereonites 1–4 org:act:9744DBAC-6A5C-4C5A-A622-D96F0A1C9F0A. subequal. Pleon with pleonite 1 wider than pleonites Etymology: Tis species is named for Dr Kerry Ann Had- 2–4, visible in dorsal view; posterior margin of pleonites feld in gratitude for her expertise in cymothoid taxon- 1–4 posteriorly concave, pleonite 5 posteriorly slightly omy and for making RL Welicky’s postdoctoral tenure in concave to straight, mostly concave. Pleonite 2 not over- South Africa a truly memorable and valuable experience lapped by pereonite 7; posterolateral angles of pleonite 2 to her both academically and personally. narrowly rounded. Pleonite 1 similar in form to pleonites 2 and 3. Pleonites 3–5 similar in form to pleonite 2; ple- Description onite 5 not extended posteriorly and equal width to ple- Female. [BMNH1952.9.9.29–30; Fig. 19.] Size onite 4, posterior margin straight. Pleotelson 0.9 times as 42.0 × 16.0. Body ovoid, weakly twisted, 2.6 times as long long as anterior width, dorsal surface with lateral indent, as greatest width, dorsal surfaces smooth and polished lateral margins weakly convex, posterior margin rounded, in appearance, widest at pereonite 5, most narrow at with medial indent. pereonite 1, pereonite lateral margins mostly posteriorly Antennula approximately as stout as antenna, com- ovate. Cephalon 0.52 as long as wide as measured in dor- prised of 8 articles; peduncle articles 1 and 2 distinct and sal view, trapezoid-shaped. Eyes oval, with distinct mar- articulated; article 2 1.3 times as long as article 1; arti- gins, one eye width 0.1 times width of cephalon; one eye cle 3 1 times as long as wide, 0.4 times as long as com- length 0.4 times length of cephalon. Pereonite 1 in line bined lengths of articles 1 and 2; fagellum with 5 articles, with base of cephalon, smooth, anterior border straight, extending to middle of eye. Antenna comprised of 9 arti- anterolateral angle narrowly rounded. Posterolateral cles; peduncle article 3 1.6 times as long as article 2; arti- margins of pereonites smooth and straight, posterolat- cle 4 1.4 times as long as wide, 0.9 times as long as article eral margin of pereonite 7 acute and extending in an arc 3; article 5 2.8 times as long as wide, 1.4 times as long as abruptly laterally and dorsally. Coxae 2–3 similar width article 4; fagellum with 4 articles, extending to middle of to coxae 4–7; with posteroventral angles rounded; coxae pereonite 1. 4–7 with rounded point and curved; not extending past Pereopod 1 basis 2.4 times as long as greatest width; posterior pereonite margin. Pereonites 1–5 increasing ischium 0.42 times as long as basis; merus proximal Welicky and Smit Parasites Vectors (2019) 12:387 Page 30 of 34

d a b e f

h i

Fig. 19 Anilocra hadfeldae n. sp. (42.0 16.0, holotype, BMNH1952.9.9.29-30). a Dorsal view. b Lateral view. c Dorsal view of cephalon. d ♀ × Pleotelson. e Antennula. f Antenna. g Pereopod 1. h Pereopod 2. i Pereopod 6. j Pereopod 7 margin without bulbous protrusion; carpus with straight dactylus 1.2 as long as propodus. Pereopods gradually proximal margin; propodus 1.5 times as long as wide; increasing in size towards posterior. Pereopod 6 basis dactylus stout, 1.4 as long as propodus, 2.4 times as long 3.2 times as long as greatest width, ischium 0.5 times as as basal width. Pereopod 2 propodus 1.7 as long as wide; long as basis, propodus 1.4 as long as wide, dactylus 1.6 Welicky and Smit Parasites Vectors (2019) 12:387 Page 31 of 34

as long as propodus. Pereopod 7 basis 2.8 times as long 4b Pleonite 1 partly concealed by pereonite 7, body 3.5 as greatest width; ischium 0.6 as long as basis, without times as long as wide, pleotelson lateral margins con- protrusions; 0.8 as long as ischium; carpus 1.6 times as vex ……………………………A. leptosoma Bleeker, 1857 long as wide, 0.7 as long as ischium, without bulbous pro- 5a Pereonites 1–3 strongly narrowed ………………………… trusion; propodus 1.9 times as long as wide, 0.8 as long ………………………………………… A. ianhudsoni n. sp. as ischium; dactylus stout, 0.9 as long as propodus, 2.9 5b Pereonites 1–3 weakly narrowed ………………………. 6 times as long as basal width. 6a Lateral margin on article 3 of antenna peduncle elon- Pleopods without setae, exopod larger than endopod. gate, extending onto lateral margin of article 4; pereo- Uropod peduncle 0.5 times as long as rami, lateral mar- pod 6 propodus not infated …A. angeladaviesae n. sp. gin without setae; rami not extending beyond pleotelson, 6b Lateral margin on article 3 of antenna peduncle not marginal setae absent, apices narrowly rounded. Endo- elongate, not extending onto lateral margin of article 4; pod apically rounded, 5 times as long as greatest width, pereopod 6 propodus with produced curvature ………7 lateral margin weakly convex, medial margin weakly con- 7a Body 2.2 times as long as greatest width; article 3 of vex, terminating with 0 setae. Exopod extending to end of antennula peduncle 1.2 times as long as wide; dense endopod or extending beyond end of endopod. chromatophores present on pereopods………………… Male. [BMNH1952.9.9.29–30; Fig. 20.] Size 30.0 × 9.0. …………………………………A. bunkleywilliamsae n. sp. Male similar to female but smaller. Body weakly twisted, 7b Body 2.4–2.6 times as long as greatest width; article ovoid, 2.9 times as long as wide. Pleopod 2 appendix 3 of antennula peduncle article 0.8 times as long as masculina with parallel margins, distally narrowed. wide; dense chromatophores absent from pereopods ……………………………………A. capensis Leach, 1818 Remarks Anilocra hadfeldae n. sp. has a distinct body shape. Te Discussion body is more parallel than ovoid and the posterior lateral Anilocra capensis has been perhaps the most utilised margins of pereonite 7 are substantially more acute than (but not necessarily correct) name for any species of round and more obviously bent upward; pereopods 7 Anilocra in South Africa and likely in other areas of have small and light pigmentation and the pleotelson has Africa, too. Tis is not surprising as only two drawings a distinct medial indentation. Te eyes are proportionally of adult female A. capensis existed before the present narrower than the other species described above. work. With few data on A. capensis and other African Te antenna and antennula of Anilocra hadfeldae n. species, it was a logical pitfall to identify species of sp. difer from that of A. capensis, the latter being stouter. Anilocra collected from South Africa and elsewhere Te antenna peduncle articles of A. hadfeldae n. sp. have in Africa as A. capensis. Tis paper provides a detailed more parallel lateral margins than those of A. angela- description of one of the original named species, and daviesae n. sp. Anilocra hadfeldae n. sp. has no nodules descriptions of six species new to science and thus will or curved lateral produced margins of the pereopods, aid in the correct identifcation of these organisms. It making it distinct from A. capensis, A. bunkleywilliamsae is important to note that our molecular fndings were n. sp., A. paulsikkeli n. sp., and A. jovanasi n. sp. unexpected as the nucleotide- and p-distance values between A. capensis and other Anilocra showed at lev- Key to the African species of Anilocra els more similar to diferences observed between genera 1a Body weakly twisted ………………………………………. 2 [24]. In order to better quantify species and genus level 1b Body straight ……………………………………………… 3 diferences within the family Cymothoidae, multi-gene 2a Posterolateral margins of pleonite 5 extended pos- phylogenetic analyses of old and new world species that teriorly, overlaying lateral margins of pleotelson are specifcally linked to correct morphological identif- …………………………………………….. A. jovanasi n. sp. cation are needed. 2b Posterolateral margins of pleonite 5 not extended posteriorly, not overlaying lateral margins of pleo- Conclusions telson, posterolateral margins of pereonite 7 acute, Te fndings of this work should greatly reduce fur- extending in an arc abruptly laterally and dorsally ther misidentifcation of A. capensis. We provide mor- ………………………………………… A. hadfeldae n. sp. phological drawings and quantitative and qualitative 3a Body elongate, pleonite 1 concealed by pereonite 7 …. 4 descriptions of this species, as well as relevant com- 3b Body ovoid, pleonite 1 not concealed by pereonite 7 …. 5 parisons among the six new species of Anilocra from 4a Pleonite 1 substantially concealed by pereonite 7; body Africa. By providing the frst molecular data for A. 3.7 times as long as wide; pleotelson lateral margins capensis, we also contribute to improving the identi- subparallel……………………………A. paulsikkeli n. sp. fcation of cryptic species via molecular analyses. We Welicky and Smit Parasites Vectors (2019) 12:387 Page 32 of 34

a b c

e g

Fig. 20 Anilocra hadfeldae n. sp. (30.0 9.0, paratype, BMNH1952.9.9.29-3). a Dorsal view. b Lateral view. c Dorsal view of cephalon. d Ventral ♂ × view of cephalon. e Pleotelson. f Pereopod 7. g Pereopod 1 also demonstrate the value of morphological data as we Acknowledgements We thank the South African Museum, Cape Town and Ichthyology collection found an ample number of morphological diferences manager, Albé Bosman, and the Natural History Museum, London and inverte- among the closely similar species described herein. In brate collections manager, Dr. Miranda Lowe, for loaning the specimens used particular, our A. capensis redescription validates the in this study. We also thank the collections team of Two Ocean Aquarium and post doc and post graduate students of the NWU-Water Research Group, Dr. name A. capensis, and provides critical baseline data O. Kudlai, D. van Rooyen and A. Greyling for feldwork assistance. We thank Dr to understand species diferences of African species of T. Sańko for his assistance in the preparation of the molecular portion of this Anilocra. manuscript. We thank Dr K. A. Hadfeld, Dr N. L. Bruce, two anonymous review- ers for their valuable feedback on the manuscript. This is contribution No. 271 from the NWU-Water Research Group. Abbreviations Authors’ contributions NHM: The Natural History Museum of London, UK; SAM, SAMC: The iZiko Both authors conceived and designed the study reported on here. RLW South Africa Museum, Cape Town, South Africa; BLASTn: Basic Local Alignment completed the data collection, morphological and molecular descriptions, Search Tool for nucleotides; cox1: mitochondrial cytochrome c oxidase subunit and write-up of the paper. NJS provided taxonomic expertise and editorial 1 gene. Welicky and Smit Parasites Vectors (2019) 12:387 Page 33 of 34

feedback on the manuscript. Both authors read and approved the fnal 10. Welicky RL, Sikkel PC. Variation in occurrence of the fsh-parasitic manuscript. cymothoid isopod, Anilocra haemuli, infecting French grunt (Haemu- lon favolineatum) in the north-eastern Caribbean. Mar Freshw Res. Funding 2014;65:1018–26. Financial assistance for RLW from the Claude Leon Foundation of South Africa 11. Folmer O, Black M, Hoeh W, Lutz R, Vrijenhoek A. DNA primers for ampli- for this research is acknowledged. The fnancial assistance of the South African fcation of mitochondrial cytochrome c oxidase subunit I from diverse National Research Foundation (NRF) towards this research is hereby acknowl- metazoan invertebrates. Mol Mar Biol Biotechnol. 1994;3:94–299. edged (NRF project IFR1702100222411, Grant 109352, NJS, PI). Opinions 12. Sudhir Kumar S, Stecher G, Tamura K. MEGA7: Molecular Evolutionary expressed and conclusions arrived at, are those of the authors and are not Genetics Analysis version 7.0. Mol Biol Evol. 2016;33:1870–4. necessarily to be attributed to the NRF. 13. Hadfeld KA, Bruce NL, Smit NJ. Redescription of the monotypic genus Cinusa Schiöedte and Meinert, 1884 (Isopoda, Cymothoidae), a buccal- Availability of data and materials cavity isopod from South Africa. Zootaxa. 2010;2437:51–68. Data supporting the conclusions of this article are included within the article. 14. Hadfeld KA, Bruce NL, Smit NJ. Review of the fsh-parasitic genus The sequences were submitted to the GenBank database under the Accession Cymothoa Fabricius, 1783 (Isopoda, Cymothoidae, Crustacea) from the Numbers MK450443-MK450452. The type-material per species is deposited in south-western Indian Ocean, including a new species from South Africa. the museum from which it was collected. Zootaxa. 2013;3640:152–76. 15. Coleman CO, Lowry JK, Macfarlane T. DELTA for beginners. An introduc- Ethics approval and consent to participate tion into the taxonomy software package DELTA. ZooKeys. 2010;45:1–75. This study received the relevant ethical approval from the North-West Univer- 16. Hadfeld KA, Bruce NL, Szinetár C, Smit NJ. Ceratothoa retusa (Schiœdte & sity Animal Ethics Committee (NWU-0044016-S5) and specimen collection by Meinert, 1883) (Isopoda, Cymothoidae), a variable species of fsh parasitic Two Oceans Aquarium was permitted by South Africa’s Department of Envi- marine isopod from the Indian Ocean. Crustaceana. 2014;87:448–62. ronmental Afairs and Department of Agriculture, Forest, and Fisheries under 17. Hadfeld KA, Bruce NL, Smit NJ. Redescription of poorly known species the integrated environmental and fsheries research and development permit of Ceratothoa Dana, 1852 (Crustacea, Isopoda, Cymothoidae), based on for the purposes of a scientifc investigation or practical experiment in terms original type material. ZooKeys. 2016;592:39–91. of Section 83 of the Marine Living Resources Act, 1998 (Act No. 18 of 1998). 18. Brandt A, Poore GCB. Higher classifcation of the fabelliferan and related isopods based on a reappraisal of relationships. Invert Syst. Consent for publication 2003;17:893–923. Not applicable. 19. Frose R, Pauly D, eds. FishBase. World Wide Web electronic publication. http://www.fshbase.org, version (06/2018). Accessed 5 Oct 2018. Competing interests 20. ICZN. International Commission on Zoological Nomenclature: Amendment The authors declare that they have no competing interests. of articles 8, 9, 10, 21 and 78 of the International Code of Zoological Nomenclature to expand and refne methods of publication. Bull Zool Author details Nomencl. 2012;69:161–9. 1 Water Research Group, Unit for Environmental Sciences and Management, 21. Bruce NL. Australian Pleopodias Richardson, 1910, and Anilocra Leach, Potchefstroom Campus, North-West University, Private Bag X6001, Potchef- 1818 (lsopoda: Cymothoidae), crustacean parasites of marine fshes. Rec stroom 2520, South Africa. 2 Present Address: School of Aquatic and Fishery Aust Mus. 1987;39:85–130. Sciences, University of Washington, 1122 NE Boat Street, Seattle, WA 98105, 22. Kussakin OG. Marine and brackish-water isopod Crustacea. Suborder USA. Flabellifera. Leningrad: Academy of Sciences USSR; 1979. 23. Ellis J. Some type specimens of isopods (Flabellifera) in the British Received: 16 August 2018 Accepted: 19 June 2019 Museum (Natural History), and the isopods in the Linnaean Collection. Bull Br Mus (Nat Hist). 1981;40:121–8. 24. Welicky RL, Hadfeld KA, Sikkel PC, Smit NJ. Molecular assessment of three species of Anilocra (Isopoda, Cymothoidae) ectoparasites from Caribbean coral reef fshes, with the description of Anilocra brillae sp. n. ZooKeys. References 2017;663:21–43. 1. Smit NJ, Bruce NL, Hadfeld KA. Global diversity of fsh parasitic isopod 25. Thatcher VE, Blumenfeld CL. Anilocra montti sp. n. (Isopoda, Cymothoi- crustaceans of the family Cymothoidae. Int J Parasitol Parasites Wldl. dae) a parasite of caged salmon and trout in Chile. Rev Brasil Zool. 2014;2:188–97. 2001;18:269–76. 2. Binning SA, Roche DG, Layton C. Ectoparasites increase swimming costs 26. Bariche M, Trilles JP. Anilocra pilchardi n. sp., a new parasitic cymothoid in a coral reef fsh. Biol Lett. 2013;9:20120927. isopod from of Lebanon (Eastern Mediterranean). Syst Parasitol. 3. Welicky RL, Demopoulos AW, Sikkel PC. Host-dependent diferences 2006;64:203–14. in resource use associated with Anilocra spp. parasitism in two coral 27. Winfeld I, Alvarez F, Ortiz M. A new species of Anilocra (Crustacea: Isop- reef fshes, as revealed by stable carbon and nitrogen isotope analyses. oda: Cymothoidae), ecto parasite on the mako shark Isurus oxyrinchus. Marine Ecol. 2017;38:e12413. Proc Biol Soc Wash. 2006;115:148–52. 4. Welicky RL, Parkyn DC, Sikkel PC. Host-dependent diferences in meas- 28. Milne-Edwards H. Histoire naturelle des Crustacés comprenent ures of condition associated with Anilocra spp. parasitism in two coral l’anatomie la physiologie et la classifcation de ces animaux. Paris: Librai- reef fshes. Env Biol Fish. 2018;101:1223–34. rie Encyclopédique de Roret; 1840. 5. Smit NJ, Hadfeld KA. Marine fsh parasitology in South Africa: history of 29. Trilles JP. Les Cymothoidae (Isopoda, Flabellifera) des collections du discovery and future direction. Afr Zool. 2015;50:79–92. Muséum National d’Histoire Naturelle de Paris. II. Les Anilocridae Schiödte 6. Leach WE. Cymothoidées. In: Cuvier F, editor. Dictionnaire des Sciences et Meinert, 1881. Genres Anilocra Leach, 1818 et Nerocila Leach, 1818. Bull Naturelle 12. Strasbourg & Paris: Levrault & Le Normant. 1818; p. 338–354. Mus Nat Hist Nat, Paris, 3e série, 290 Zool. 1975;200:303–40. 7. Schiodte JC, Meinert FR. Symbolae ad monographium cymothoarum 30. Bruce NL, Harrison-Nelson EB. New records of fsh parasitic marine isopod crustaceorum familiae. Ill. Saophridae. IV. Ceratothoinae. Naturhistorisk crustaceans (Cymothoidae, subfamily Anilocrinae) from the Indo-West Tidsskrift. 1883;13:281–378. Pacifc. Proc Biol Soc Wash. 1988;101:585–602. 8. Williams LB, Williams EH. Nine new species of Anilocra (Crustacea: 31. Wright RV, Lechanteur YARG, Prochazka K, Grifths CL. Infection of hot- Isopoda: Cymothoidae) external parasites of West Indian coral reef fshes. tentot Pachymetopon blochii by the fsh louse Anilocra capensis (Crusta- Proc Biol Soc Wash. 1981;94:1005–47. cea: Isopoda) in False Bay, South Africa. Afr Zool. 2001;36:177–83. 9. Bruce NL. The genera Catoessa, Elthusa, Enispa, Ichthyoxenus, Idusa, 32. Thorsen DH, Trilles JP. The occurrence of Anilocra capensis and Nerocila Livoneca and Norileca n. gen. (Isopoda, Cymothoidae), crustacean para- armata (Isopoda: Cymothoidae) in the Canary Islands with comments on sites of marine fshes, with descriptions of eastern Australian species. Rec their novel hosts. Bull Mar Sci. 2002;70:227–31. Aust Mus. 1990;42:247–300. Welicky and Smit Parasites Vectors (2019) 12:387 Page 34 of 34

33. Thorsen DH, Mille KJ, Van Tassell JL, Hajagos JG. Infestation of the par- 35. Monod T. Parasitologia Mauritanica. Materiaux pour la faune parasi- rotfsh Sparisoma cretense (Scaridae), by the fsh louse, Anilocra physodes tologique en Mauretanie. Fascicule 1, Deuxième partie. Isopoda. Bull Com (Isopoda: Cymothoidae), in the Canary Islands. Cybium. 2002;24:45–59. d’Etude Histor Sci Afri Occident Fr. 1924;9:73–4. 34. Aneesh PT, Helna AK, Trilles JP, Chandra K. Occurrence and redescription of Anilocra leptosoma Bleeker, 1857 (Crustacea: Isopoda: Cymothoidae) parasitizing the clupeid fsh Tenualosa toli (Valenciennes) from the Publisher’s Note Arabian Sea, India. Mar Biodiv. 2017. https://doi.org/10.1007/s1252 Springer Nature remains neutral with regard to jurisdictional claims in pub- 6-017-0828-7. lished maps and institutional afliations.

Ready to submit your research ? Choose BMC and benefit from:

• fast, convenient online submission • thorough peer review by experienced researchers in your ﬁeld • rapid publication on acceptance • support for research data, including large and complex data types • gold Open Access which fosters wider collaboration and increased citations • maximum visibility for your research: over 100M website views per year

At BMC, research is always in progress.

Learn more biomedcentral.com/submissions