Fig 1.
Female of Simulium mellah (Spain, Turrillas, 24 June 2018).
(a) Terminalia, ventral view; a, anteriorly directed apodeme of lateral arm of genital fork; al, anal lobe; c, cercus; f, stem of genital fork; sp, spermatheca; v, hypogynial valve; IX, tergite of abdominal segment IX; VIIIs and VIIIt, sternite and tergite, respectively, of abdominal segment VIII; (b) Genital fork, ventral view; (c) Right anal lobe and cercus, ventral view; (d) Spermatheca; arrow indicates pigmented extension onto base of spermathecal duct; (e) Abdominal segment VIII with hypogynial valves, ventral view; (f) Claw of hind leg; (g) Maxillary palp; segments I–IV only (V not shown); (h) Apex of mandible.
Fig 2.
Male of Simulium mellah (Spain, El Cautivo, 21 May 2019).
(a) Terminalia, terminal view; ae, aedeagal membrane; c, cercus; d, dorsal plate; gc, gonocoxa; gs, gonostylus; m, median sclerite; p, paramere; vp, ventral plate; X, tergite of abdominal segment X; (b) Genitalia, ventral view; (c) Right gonocoxa and gonostylus, inner lateral view; (d) Aedeagal membrane (ae), dorsal plate (d), and paramere (p) with parameral spine; terminal view; (e) Ventral plate and median sclerite, terminal view; (f) Gonostylus, dorsal view. G. Ventral plate, dorsal view; (h) Ventral plate, ventral view; (i) Ventral plate, lateral view.
Fig 3.
Larva of Simulium mellah (Spain, El Cautivo, 7 April 2020).
(a, b) Head capsule, dorsal view; (c, d) Head capsule, ventral view; (e, f) Hypostoma; (a, c, e) Light form; (b, d, f) Dark form.
Fig 4.
Pupa and cocoon of Simulium mellah (Spain, El Cautivo, 21 May 2019).
(a, b) Gill base, showing variation, lateral view; (c) Middle of cephalic plate (male); (d) Anterior margin of cocoon, lateral view; (e) Cocoon, lateral view; box indicates area enlarged in d.
Table 1.
Diagnostic characters for Simulium mellah versus other species of the S. aureum group in Spain.
Fig 5.
IIL chromosomal sequence of Simulium mellah (female larva, Spain, El Cautivo, 13 November 2019), showing fixed inversion IIL-20 and limits of polymorphic inversion IIL-52.
Table 2.
Frequency of chromosomal rearrangements in larvae of Simulium mellah from Spain, Almería province, Tabernas, El Cautivo.
Fig 6.
(a) Phylogenetic tree based on COI sequences, with maximum likelihood topology, of black flies from southeastern Spain, together with sequences from GenBank. The bootstrap values are only shown on nodes with bootstrap values higher than 75%, indicated by black circles in the figure, and range from 75% to 100% depending on their size. (b) Zoom-in of the group formed by S. rubzovianum and S. mellah. Unknown individuals correspond to larvae of subgenus Eusimulium which were not identified to species level prior to DNA analysis. Clusters with a value in brackets at the end of the species mean that these sequences have been collapsed and the information on the component species is in S1 Table. All sequences used for the phylogeny are summarized in S1 Table.
Fig 7.
Phylogenetic tree based on ITS2 sequences, with maximum likelihood topology, of black flies from southeastern Spain, together with sequences from GenBank.
The bootstrap values are only shown on nodes with bootstrap values higher than 75%, indicated by black circles in the figure, and range from 75% to 100% depending on their size. The unknown larvae were assigned to the closest group due to the high resolution and congruence shown by ITS2. Clusters with a value in brackets at the end of the species mean that these sequences have been collapsed and the information on the component species is in S2 Table. All sequences used for the phylogeny are summarized in S2 Table.
Fig 8.
Boxplot of minimum interspecific and maximum intraspecific genetic distances per sequence from all COI and ITS2 sequences of black flies.
The median is indicated by the line inside the box. The lower and upper hinges correspond to the first and third quartiles (the 25th and 75th percentiles). The upper whisker extends from the hinge to the largest value no farther than 1.5 * IQR from the hinge (where IQR is the inter-quartile range, or distance between the first and third quartiles). The lower whisker extends from the hinge to the smallest value, at most 1.5 * IQR of the hinge. Data beyond the end of the whiskers are outlying points and are plotted individually.
Table 3.
Mean and ranges of intra- and interspecific distances per species and DNA region and identification status of black flies from southeastern Spain, based on “best close match”.
For COI sequences, given a threshold of 0.6% obtained by “threshOpt” and a threshold of 2.36% based on TaxonDNA. For ITS2 sequences, given a threshold of 0.92% obtained by “threshOpt” and a threshold of 0.96% based on TaxonDNA.