New and cryptic species of intertidal mites (Acari, Oribatida) from the Western Caribbean – an integrative approach

Figures & data

Table 1. GenBank accession numbers for COI and 18S rRNA gene sequences comprising all specimens included in genetic investigations. Sequences generated in this study appear in bold.

			GenBank Accession Nr.
Species	Sample ID	Coordinates	COI	18S
Thalassozetes balboa sp. nov.	T_PA_37_3	9.415057°N,82.330787°W		MK035018
	T_PA_43_3	9.370821°N,82.239908°W		MK035019
Thalassozetes barbara	T_BA_30_1	13.212719°N,59.517116°W		MK035020
Litoribates floridae sp. nov.	L_FL_08_3	24.914264°N, 80.64089°W	MK035001	MK035021
	L_FL_08_4		MK035002
	L_FL_08_6		MK035003
	L_FL_08_7		MK035004
	L_FL_08_8		MK035005
	L_FL_13_1	24.890059°N,80.674575°W	MK035006	MK035022
Litoribates bonairensis	L_BO_01_1	12.103654°N,68.221720°W	MK035007	MF997503^d
	L_BO_01_2		MK035008	MF997502^d
	L_BO_01_4		MK035009
	L_BOa_01_1		MK035010
	L_BO3_02_1		MK035011
	L_BO3_02_1^a		MK035012
	L_BOa_01_11		MK035013
	L_BOa_01_12		MK035014
	L_BOa_01_8		MK035015
	L_BOa_01_9		MK035016
	L2_BO_01a_1		MK035017
Alismobates pseudoreticulatus	A_MY_07_7		MH285613^a	MH285696^a
	A_MY_05_3		MH285597^a
	A_MY_07_4		MH285610 ^a
Alismobates reticulatus				AB818526.1^b
Aquanothrus sp.				KX397627.1^c
Fortuynia longiseta				MH285693^a
Fortuynia rotunda				AB818525.1^b
Fortuynia sp.				MH285695^a
Fortuynia smiti				MH285694^a
Hydrozetes confervae				AB818523.1^b
Hydrozetes lemnae				KX397632.1^c
Indopacifica pantai (1)				MH285692^a
Indopacifica pantai (2)				MH285691^a
Indopacifica parva				MH285690^a
Schusteria littorea				HM070345.1^e
Thasecazetes falcidactylus				MF997501^d
Thalassozetes shimojanai				AB818524.1^b

^aPfingstl et al. (2018); ^bIseki & Karasawa (2014); ^cKrause et al. (2016); ^dPfingstl et al. (2017); ^ePepato et al. (2010).

Figure 1. Maps illustrating the records of new species in the western Caribbean. Left vertical map provides an overview of the area with white frames indicating the position of the regions given in the two horizontal maps. Red circles represent records of Thalassozetes balboa sp. nov. and orange circles identify occurrence of Litoribates floridae sp. nov.; codes in white refer to the sampling location given in the Materials and Method section.

Figure 2. Graphic illustration of measured continuous variables shown on simplified drawings of Thalassozetes barbara. (a) Dorsal aspect: bl – body length, dPtI – distance between pedotecta I, db – distance between bothridia, ll – lenticulus length, dnr – distance notogastral ridges, nw_da – notogastral width on level of seta da, nw_dm – notogastral width on level of seta dm, nw_dp – notogastral width on level of seta dp. (b) Ventral aspect: cl – camerostome length, cw – camerostome width, dre1 – distance between ridges on epimeron 1, efl – epimeral fovea length, efw₁ – epimeral fovea width anterior part, efw₂ – epimeral fovea width posterior part, dcg – distance between camerostome and genital orifice, dac3 – distance between acetabula 3, gl – genital orifice length, gw – genital orifice width, al – anal opening length, aw – anal opening width.

Figure 3. Litoribates floridae sp. nov. adult (a) dorsal view; (b) ventral view, distal leg segments omitted; (c) lateral view.

Figure 4. Photographs (stacked stereomicroscopic images) of Litoribates floridae sp. nov. adult (a) dorsal view; (b) ventral view; (c) lateral view. Scale bars 100 µm.

Figure 5. Litoribates floridae sp. nov. adult mouthparts (a) right rutellum, ventral view; (b) left pedipalp, paraxial view; (c) left chelicera, antiaxial view.

Table 2. Leg chaetome and solenidia for the new species.

	Trochanter	Femur	Genu	Tibia	Tarsus	Chaetome	Solenidia
Litoribates floridae sp. nov.
Leg I	–	d, bv´´, (l)	(l), σ	(l), v´´, φ1, φ2	(pl), (pv), s, (a), (u), (p), (tc), (ft), (it), ε, ω1, ω2	0-4-2-3-18	1-2-2
Leg II	–	d, bv´´, (l)	(l), σ	(l), v´, φ	(pv), s, (a), (u), (p), (tc), (ft), (it), ω	0-4-2-3-15	1-1-1
Leg III	v´	d, l´, ev´	l´, σ	(l), v´´, φ	(pv), s, (a), (u), (p), (tc), (ft), (it)	1-3-1-3-15	1-1-0
Leg IV	v´	d, ev´	d, l´	l´, v´, φ	(pv), s, (a), (u), (p), (tc), ft´´	1-2-2-2-12	0-1-0
Thalassozetes balboa sp. nov.
Leg I	–	d, (l)	(l), σ	(l), v´, φ1, φ2	(pl), (pv), s, (a), (u), (p), (tc), (ft), (it), ε, ω1, ω2	0-3-2-3-18	1-2-2
Leg II	–	d, (l)	(l), σ	(l), v´, φ	(pv), s, (a), (u), (p), (tc), (ft), (it), ω	0-3-2-3-15	1-1-1
Leg III	v´	d, ev´	l´, σ	l´, v´´, φ	(pv), s, (a), (u), (p), (tc), (ft)	1-2-1-2-13	1-1-0
Leg IV	v´	d, ev´	l´	l´, (v), φ	(pv), s, (a), (u), (p), (tc), ft´´	1-2-1-3-12	0-1-0

() = pairs of setae.

Figure 6. Litoribates floridae sp. nov. adult left legs, antiaxial view (a) leg I; (b) leg II; (c) leg III; (d) leg IV.

Figure 7. Thalassozetes balboa sp. nov. adult (a) dorsal view; (b) ventral view, distal leg segments omitted; (c) lateral view.

Figure 8. Photographs (stacked stereomicroscopic images) of Thalassozetes balboa sp. nov. adult (a) dorsal view; (b) ventral view; (c) lateral view. Scale bars 100 µm.

Figure 9. Thalassozetes balboa sp. nov. adult mouthparts (a) right rutellum, ventral view; (b) left pedipalp, paraxial view; (c) left chelicera, antiaxial view.

Figure 10. Thalassozetes balboa sp. nov. adult left legs, antiaxial view (a) leg I; (b) leg II; (c) leg III; (d) leg IV.

Table 3. Univariate statistics for Caribbean Litoribates species.

	L. bonairensis (n = 19)					L. floridae sp. nov. (n = 16)
	min	max	x	sd	cv	min	max	x	sd	cv	Mann–Whitney-U
bl	345	382	364.21	8.97	0.02	350	375	363.31	7.88	0.02
dPtI	151	163	157.95	3.75	0.02	160	166	162.19	2.23	0.01	***
db	132	148	138.11	4.98	0.04	139	148	142.31	3.09	0.02	**
ll	46	65	51.95	5.16	0.10	52	65	59.44	4.16	0.07	***
nwc1	160	194	176.00	9.33	0.05	163	203	178.50	10.71	0.06
nwda	228	252	239.58	7.83	0.03	231	259	244.25	8.63	0.04
nwdm	219	259	237.84	10.54	0.04	222	259	237.13	9.49	0.04
cl	89	108	99.58	5.63	0.06	92	108	100.06	4.57	0.05
cw	80	89	84.53	2.41	0.03	83	86	83.94	1.44	0.02
dcg	71	92	81.74	5.86	0.07	68	92	79.31	6.29	0.08
dac3	129	142	135.47	3.66	0.03	135	142	138.63	3.14	0.02	*
gl	52	65	58.47	3.63	0.06	53	65	59.50	3.46	0.06
gw	59	72	64.53	3.10	0.05	60	72	66.44	4.35	0.07
al	80	86	81.26	1.82	0.02	80	86	83.81	1.94	0.02	***
aw	65	74	69.63	2.56	0.04	65	74	69.63	3.24	0.05

Minimum (min), maximum (max), mean (x), standard deviation (sd), and coefficient of variation (cv), marked grey if equal or higher than 0.10. Results of Mann–Whitney-U test are given; *p < 0.05, **p < 0.01, ***p < 0.001.

Table 4. Loadings of the two principal components PC 1 and PC 2 gained from PCA on data of L. bonairensis and L. floridae sp. nov.

	Raw data		Size-corrected data
	PC 1	PC 2	PC 1	PC 2
bl	0.09	0.15	0.14	0.12
dPtI	0.13	0.10	0.03	−0.08
db	0.17	0.16	0.05	−0.09
ll	0.79	−0.44	−0.67	−0.11
nwc1	0.27	0.08	−0.37	−0.18
nwda	0.20	0.18	−0.03	0.13
nwdm	0.06	0.29	0.39	0.41
cl	0.18	0.12	−0.03	0.38
cw	0.03	0.12	0.12	0.00
dcg	−0.11	0.37	0.42	−0.72
dac3	0.11	0.15	0.08	−0.11
gl	0.21	0.49	0.12	0.15
gw	0.30	0.35	−0.03	0.19
al	0.11	−0.08	−0.08	0.03
aw	0.07	0.27	0.14	0.03

High loadings explaining differences between the species are given in bold.

Figure 11. Scatter plots of PCA of L. bonairensis from Bonaire and L. floridae sp. nov. from Florida. Open symbols refer to females; filled symbols refer to male specimens. For better display of sexual dimorphism, each gender is given in a separate hull.

Table 5. Univariate statistics for Caribbean Thalassozetes species.

	T. barabara (n = 19)					T. balboae (n = 90)
	min	max	x	sd	cv	min	max	x	sd	cv	Mann–Whitney-U
bl	271	302	284.68	8.63	0.03	277	312	292.58	9.34	0.03	**
dPtI	120	129	124.42	2.09	0.02	117	135	126.62	3.19	0.03	**
db	49	53	51.26	1.33	0.03	51	59	54.98	2.25	0.04	***
ll	28	34	30.53	2.29	0.08	31	46	38.97	3.61	0.09	***
dnr	31	43	36.58	2.65	0.07	28	40	34.00	2.84	0.08	***
nwda	154	169	158.89	5.02	0.03	142	182	161.86	7.33	0.05
nwdm	163	182	170.32	5.62	0.03	154	194	173.50	7.17	0.04
nwdp	145	169	154.37	5.79	0.04	142	175	156.83	6.27	0.04
cl	74	83	77.16	2.54	0.03	70	83	78.58	2.50	0.03	*
cw	55	62	59.74	1.82	0.03	55	62	59.54	1.87	0.03
dre1	105	114	110.68	2.43	0.02	108	123	115.67	2.96	0.03	***
efl	23	31	27.58	1.64	0.06	25	31	28.17	1.78	0.06
efw1	22	31	25.47	2.29	0.09	19	30	23.78	2.04	0.09	**
efw2	19	25	21.11	2.28	0.11	14	22	16.57	2.07	0.13	***
dcg	68	77	72.32	2.50	0.03	69	81	75.30	2.41	0.03	***
dac3	92	105	96.11	2.92	0.03	92	108	99.83	3.23	0.03	***
gl	34	45	38.32	3.77	0.10	34	46	38.46	3.56	0.09
gw	42	49	45.37	2.99	0.07	40	55	45.79	3.71	0.08
al	62	68	63.26	1.82	0.03	60	71	64.16	2.16	0.03
aw	46	52	49.11	1.79	0.04	43	55	48.90	2.23	0.05

Minimum (min), maximum (max), mean (x), standard deviation (sd), and coefficient of variation (cv), marked grey if equal or higher than 0.10. Results of Mann–Whitney-U test are given; *p < 0.05, **p < 0.01, ***p < 0.001.

Table 6. Loadings of the two principal components PC 1 and PC 2 gained from PCA on data of T. barbara and T. balboa sp. nov.

	Raw data		Size-corrected data
	PC 1	PC 2	PC 1	PC 2
bl	−0.03	0.17	0.11	0.02
dPtI	−0.02	0.11	0.07	−0.11
db	−0.12	0.15	0.20	−0.04
ll	−0.57	0.30	0.73	0.14
dnr	0.16	0.18	−0.21	−0.13
nwda	−0.01	0.23	0.03	0.20
nwdm	−0.01	0.23	0.03	0.20
nwdp	0.00	0.19	0.01	0.07
cl	−0.03	0.06	0.08	−0.19
cw	0.03	0.10	−0.04	−0.13
dre1	−0.07	0.13	0.16	−0.06
efl	−0.02	−0.01	0.04	−0.28
efw1	0.22	0.17	−0.20	−0.05
efw2	0.75	0.17	−0.46	−0.15
dcg	−0.09	0.03	0.21	−0.31
dac3	−0.06	0.13	0.12	−0.06
gl	0.02	0.54	−0.10	0.57
gw	0.02	0.46	−0.08	0.51
al	−0.02	0.14	0.04	−0.05
aw	0.04	0.19	−0.05	0.06

High loadings explaining differences between the species are given in bold.

Figure 12. Scatter plots of PCA of T. barbara from Barbados and T. balboa sp. nov. from three different locations in Panama. Codes refer to sample sites given in the Materials and Method section; open symbols refer to females; filled symbols refer to male specimens.

Figure 13. Bayesian inference tree of three fortuyniid species based on COI sequences constructed by means of MrBayes applying the GTR+I+G model. Posterior probabilities for main nodes are shown above branches. Sequences obtained from GenBank are marked by *. For details, see Table 1. ** Mean of intra- (blue, red) and interspecific (violet) uncorrected p-distances given in per cent.

Figure 14. Bayesian inference tree of fortuyniid and selenoribatid taxa based on 18S rRNA sequences (1811 bp) constructed by means of MrBayes applying the GTR+I+G model. Posterior probabilities (>80) are shown near nodes. Sequences obtained from GenBank are marked by *. For details, see Table 1.

Figure 15. Photographs (stacked stereomicroscopic images) of female adult Litoribates specimens used for morphometric investigations. (a) Litoribates floridae sp. nov., Florida; (b) Litoribates bonairensis, Bonaire. Scale bars 100 µm.

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