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Figure 7 in First records of two non-indigenous tineid species from St Helena Island (Tineioidia: Tineidae)

Figure 7 – Opuntia plants and former garden land, an example of habitat for A. siccata on St Helena (T. Fowler)

Figure 9 in First records of two non-indigenous tineid species from St Helena Island (Tineioidia: Tineidae)

Figure 9 – Habitat of S. rutella in Cole's courtyard, Jamestown: ensemble of buildings and walls with a Mango tree in the middle (T. Karisch)

Figures 4 & 5 – A. siccata, gen. slide 3937. 4a in First records of two non-indigenous tineid species from St Helena Island (Tineioidia: Tineidae)

Figures 4 & 5 – A. siccata, gen. slide 3937. 4a: ♂-genitalia, 4b: aedeagus. – A. siccata, gen. slide 3944. 5a: ♀-genitalia, 5b: signum (lateral view).

Figures 1–3 in First records of two non-indigenous tineid species from St Helena Island (Tineioidia: Tineidae)

Figures 1–3 Adults: Amphixystis siccata 1. ♂ (St. Helena); 2. A. siccata ♂ Lectotype (Mauritius); 3. Setomorpha rutella ♂ (St. Helena)

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Fig. 1 in New Alien Species Mytilopsis Leucophaeata And Corbicula Fluminalis (Mollusca, Bivalvia) Recorded In Georgia And Notes On Other Non-Indigenous Molluscs Invaded The South Caucasus

Fig. 1. Map of the region showing the collection sites. Details for each sampling point are given in table 1.

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Fig. 3 in New Alien Species Mytilopsis Leucophaeata And Corbicula Fluminalis (Mollusca, Bivalvia) Recorded In Georgia And Notes On Other Non-Indigenous Molluscs Invaded The South Caucasus

Fig. 3. Corbicula fluminalis: 1–4 whole specimen from native range, Vilesh River near Masally, Azerbaijan (1, 2 — right valve, 3, 4 — left valve); 5–8 — whole specimen from Pichori village (locality 1) (5, 6 — right valve, 7, 8 — left valve); 9–10 — a single left valve from Tbilisi Reservoir (locality 4) from outside (9) and inside (10); 11–12 — a single left valve from Shaori Reservoir (locality 3) from outside (11) and inside (12); 13–14 — a single left valve from Iori River (locality 5) from outside (13) and inside (14).

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Fig. 2 in New Alien Species Mytilopsis Leucophaeata And Corbicula Fluminalis (Mollusca, Bivalvia) Recorded In Georgia And Notes On Other Non-Indigenous Molluscs Invaded The South Caucasus

Fig. 2. Mytilopsis leucophaeata: 1 — shells from Patara Paliastomi Lake (locality 2); 2–4 — whole specimen from the same locality (2 — right valve, 3 — left valve, 4 — enlarged inner view of position 2 showing the apophysis); 5–6 — a single right valve of shell from Shaori Reservoir (locality 3) from outside (5) and inside (6) views correspondingly.

Fig. 6 in Polyclinum constellatum (Tunicata, Ascidiacea), an emerging non-indigenous species of the Mediterranean Sea: integrated taxonomy and the importance of reliable DNA barcode data Abstract

Fig. 6: ML phylogenetic tree of the genus Polyclinum (sequences abbreviation: Pln) based on COI nucleotide sequences (1560 aligned nucleotide sites; best-fit substitution model GTR+I+G; bootstrap on 100 replicates). Eudistoma and Pseudodistoma species were used as outgroups. The sequence list and species abbreviations are reported in Supplementary table S1. Black dots: bootstrap values ≥ 70 %; red: P. constellatum sequences; blue: P. indicum sequences; yellow background: our sequences.

Fig. 4 in Polyclinum constellatum (Tunicata, Ascidiacea), an emerging non-indigenous species of the Mediterranean Sea: integrated taxonomy and the importance of reliable DNA barcode data Abstract

Fig. 4: A, C) Colonies of Polyclinum constellatum with different colours photographed and collected in the Heraklion marina (Crete) (A: colony K11 and C: colony K12); B) Transversal section of the colonies, joined only at the surface layer (upper white arrow); D) Zooid extracted from the red-orange colony (K11), with magnification of the 6-lobed anus; E) Zooid extracted from the dark blue colony (K12) with magnification of the 6-lobed anus. Both K11 and K12 have the same COI haplotype (sequence AC number: MT873559).

Fig. 5 in Polyclinum constellatum (Tunicata, Ascidiacea), an emerging non-indigenous species of the Mediterranean Sea: integrated taxonomy and the importance of reliable DNA barcode data Abstract

Fig. 5: A) Larva of P. constellatum, showing the ocellus, four long narrow ampullae, three adhesive papillae and a group of a few small ventral vesicles (red arrow). am, ampullae; ap, adhesive papillae; oc, ocellus; B) Larva of P. constellatum, red arrow pointing out the calcite crystal in the middle of the body.

Fig. 2 in Polyclinum constellatum (Tunicata, Ascidiacea), an emerging non-indigenous species of the Mediterranean Sea: integrated taxonomy and the importance of reliable DNA barcode data Abstract

Fig. 2: A) Orange colony of Polyclinum constellatum from Taranto harbour (colony P1); B) Magnification of the oral (arrow pointing put the oral tentacles of different size) and cloacal aperture (asterisk); C) P. constellatum collected in Heraklion (colony K19) with zooids arranged in systems around the cloacal apertures; D) Section of the colony showing the zooids located only around the outer edge (arrow).

Fig. 3 in Polyclinum constellatum (Tunicata, Ascidiacea), an emerging non-indigenous species of the Mediterranean Sea: integrated taxonomy and the importance of reliable DNA barcode data Abstract

Fig. 3: A) Whole zooid of Polyclinum constellatum, showing a clear division into thorax, abdomen and post-abdomen with a long vascular stolon. ab, abdomen; pa, post-abdomen; th, thorax; vs, vascular stolon; B) Zooid with evident pharynx, rectum, anus and four embryos incubated in the atrial cavity. The funnel-shaped oesophagus, the smooth stomach and the twisted gut loop are visible in the abdomen. The post-abdomen shows the heart at its terminal end, as well as several rounded testicular follicles and the ovary, with the gonoducts running parallel to the rectum. an, anus; e, embryos; gd, gonoducts; gl, gut loop; oe, oesophagus; ov, ovary; h, heart; r, rectum; st, stomach; tf, testicular follicles; C) Magnification of the oral siphon with six pointed lobes (arrows) and six longitudinal muscle bands (indicated with numbers 1-6); D) Branchial sac with 18 rows of stigmata and narrow languets of the dorsal lamina (arrows); E) Magnification of the pharynx, with minute papillae (arrows) at the level of the transverse vessels; F) Magnification of the six-lobed anus (lobes indicated with numbers 1-6).

Fig. 1 in Polyclinum constellatum (Tunicata, Ascidiacea), an emerging non-indigenous species of the Mediterranean Sea: integrated taxonomy and the importance of reliable DNA barcode data Abstract

Fig. 1: Map of the Mediterranean Sea showing the literature records (black rhombuses) of P. constellatum and the new findings (red dots). performed in a final reaction volume of 25 μl contain- nus was reconstructed with the online software PHYML ing: 1X reaction buffer with 1 mM final concentration of v3.0 (http://www.atgc-montpellier.fr/phyml-sms/) (Guin- MgCl 2 (Takara Bio Inc.), 0.2 mM of each dNTP, 0.3 μM don & Gascuel, 2003), which also includes the automatof each primer and 1.25 Units of PrimeStar HS (Takara ic model selection algorithm SMS (Smart Model Selec- Bio Inc.). Amplification conditions were: 30 cycles with tion). The best-fit substitution model was selected using denaturation for 10 s at 98°C, annealing for 15 s at 46°C the Akaike Information Criterion (AIC). Bootstrap val- or 50°C, extension for 1 min 30 s at 72°C; a final elonga- ues, indicating node reliability, were based on 100 reption step of 5 min at 72°C. licates. The sequence dataset used for this phylogenetic PCRs with the DreamTaq polymerase were performed reconstruction is reported in Supplementary Table S1 and in a final volume of 25 μl containing: 1X reaction buffer was extracted from the phylogenetic dataset published in with 2 mM final concentration of MgCl 2 (Thermo Fish- Tabudravu et al. (2019). It includes representative species er Scientific), 0.2 mM of each dNTP, 0.4 μM of each of of the Polyclinidae family plus Eudistoma and Pseudodithe two primers, and 1.25 Units of DreamTaq polymerase stoma species chosen as outgroups for their morphologi- (Thermo Fisher Scientific). The amplification conditions cal similarities with Polyclinidae. were as follows: an initial denaturation for 3 min at 95°C, then 34 amplification cycles (denaturation for 30 s at 95°C; annealing for 30 s at 46-50°C; extension for 1 min Results 30 s at 72°C) followed by a final elongation step of 5 min at 72°C. Morphological analyses The obtained amplicons were purified with the DNA Clean&Concentrator kit (Zymo Research) and directly The colonies collected in Taranto harbour and Hersequenced according to the Sanger method by Microsynth aklion marina were all morphologically identified as P. AG (Switzerland). The sequence quality check, compar- constellatum based on the following features: colonies isons and alignment were carried out with Geneious ver. without sand in/outside, zooids arranged in systems, 5.5.7.2 (Kearse et al., 2012). The sequences obtained post-abdomen (without vascular stolon) shorter than the were deposited in the GenBank database (see Accession thorax and abdomen combined, pharynx with 16-18 rows numbers MT873559 and OL597608). For comparative of stigmata, more than 15 stigmata per row, and a 6-lobed analyses, homologous sequences of the genus Polycli- anus. These characteristics are in accordance with the key num were searched for in the non-redundant nucleotide of Polyclinum species edited by Kott (1963) and they are database (nr-nt db, on 21st September 2021) of the NCBI also reported in the description of the species made by (National Center for Biotechnology Information) by En- Van Name (1945). trez text search, and by BLASTn (Altschul et al., 1990) using our P. constellatum sequences as the query. Uncorrected pairwise distances were calculated with PAUP 4.0a (Swofford, 2002), while a Maximum Likelihood (ML) phylogenetic tree of the genus Polyclinum ge-

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Figure 4 in The marine live bait trade as a pathway for the introduction of non-indigenous species into California: patterns of importation and thermal tolerances of imported specimens

Figure 4. Average percent survival (± SE) of G. dibranchiata (A) and Perinereis sp. (B) after exposed to southern California thermal conditions for five days. Gray bars indicate significant difference. No difference in survival was observed among the three temperature treatments for G. dibranchiata (F2,8 = 2.67, p = 0.130). There was a significant difference in survival of Perinereis sp. among the treatments (F2,8 = 11.08, p = 0.005).

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Figure 1 in The marine live bait trade as a pathway for the introduction of non-indigenous species into California: patterns of importation and thermal tolerances of imported specimens

Figure 1. Distribution by county of bait shops selling marine live bait in California according to survey responses. Live marine bait was sold in all counties shaded in grey. Numbers inside black circles indicate species sold in that county. Bait shops in shaded counties without black circles reported marine live bait sales but did not provide information on which species they sold.

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Figure 3 in The marine live bait trade as a pathway for the introduction of non-indigenous species into California: patterns of importation and thermal tolerances of imported specimens

Figure 3. For each species of marine live bait, (A) the percentage of respondents who reported importing that species who answered that either seaweed (sometimes in combination with newspaper) or seawater were used as packing materials, and (B) the percentage of respondents who reported importing that species who observed hitchhikers in shipments of that species. Note that for (A), some respondents indicated that both seaweed and seawater were used as packing materials for a given species, so that the total percentage may add up to greater than 100 (e.g., for pileworms).

Fig. 1 in Polyclinum constellatum (Tunicata, Ascidiacea), an emerging non-indigenous species of the Mediterranean Sea: integrated taxonomy and the importance of reliable DNA barcode data Abstract

Fig. 1: Map of the Mediterranean Sea showing the literature records (black rhombuses) of P. constellatum and the new findings (red dots).