Marked differentiation in a new species of dwarf stonebasher, Pollimyrus cuandoensis sp. nov. (Mormyridae: Teleostei), from a contact zone with two sibling species of the Okavango and Zambezi rivers Author Kramer, Bernd University of Regensburg, Zoological Institute, D- 93040 Regensburg, Germany; Author Bank, Herman van der Department of Zoology, University of Johannesburg, African Centre for DNA Barcoding, Johannesburg, South Africa; Author Wink, Michael Institute of Pharmacy and Molecular Biotechnology, Heidelberg University, D- 69120 Heidelberg, Germany text Journal of Natural History 2013 J. Nat. Hist. 2013-12-04 48 7 - 8 429 463 http://dx.doi.org/10.1080/00222933.2013.807950 journal article 7021 10.1080/00222933.2013.807950 a1053c94-58d2-4e68-80f8-fc983d11ee88 1464-5262 4631571 FB644B21-04F6-44F8-AEB6-8DC09268C17C Pollimyrus petricolus (Daget, 1954) GenBank accession no. AF201608 (Sullivan et al. 2000) . Appendix 2. Morphological and genetic differences between samples Table 2.1. Morphological measures for samples of the Pollimyrus castelnaui / marianne species complex from various origins. For abbreviation of morphological characters, see Material and Methods.

PDL /	PAL /	LD /	LA /	LPF /	PPF /	pD / SL	CPL /	CPD /	LS(o) /	OD /	HL /	HL / Na	BD /	nD	nA	SPc	SLS	SL (cm)
SL	SL	SL	SL	HL	SL	SL	CPL	HL	HL	SL	SL
Zambezi (Z1,Z2)
Mean / median	0.6310	0.5764	0.1738	0.2292	0.8990	0.1529	0.4136	0.2285	0.3124	0.4281	0.2023	0.2302	10.867	0.2607	17	23	16	55	6.2
Min	0.6111	0.5497	0.1486	0.1643	0.8053	0.1350	0.2802	0.1973	0.2573	0.3780	0.1677	0.2115	8.8	0.2199	14	21	13	50	3.4
Max	0.6778	0.6206	0.2009	0.2587	1.0137	0.1796	0.4983	0.2585	0.3923	0.4790	0.2246	0.2741	13.083	0.2893	19	26	18	59	7.0
SE / SIQ	0.0016	0.0018	0.0014	0.0018	0.0060	0.0012	0.0034	0.0015	0.0034	0.0028	0.0016	0.0013	0.1144	0.0018	0.5	1	0	1.5	0.09
N	64	64	64	64	64	64	64	64	64	64	64	64	64	64	63	64	62	64	64
Kwando: Kongola (K)
Mean / median	0.6516	0.6014	0.1636	0.216	0.8319	0.1696	0.395	0.2181	0.3474	0.4517	0.2635	0.2422	9.0188	0.2753	17	23	14	50	4.6
Min	0.6333	0.5849	0.14	0.18	0.7639	0.1507	0.3796	0.1898	0.3214	0.4059	0.2376	0.2255	8.3125	0.2467	15	21	12	44	3.0
Max	0.6852	0.6343	0.1818	0.2417	0.9257	0.1873	0.426	0.24	0.4146	0.5	0.3050	0.2633	9.9167	0.3051	18	24	16	55	5.7
SE / SIQ	0.0029	0.003	0.0022	0.0031	0.0139	0.002	0.0028	0.003	0.0061	0.0066	0.005	0.0024	0.1204	0.0035	0.5	0.5	0	1.5	0.153
N	19	19	19	19	15	19	19	19	19	19	19	19	19	19	19	19	18	19	19
Kwando: Nakatwa (N)
Mean / median	0.6466	0.5924	0.1540	0.2030	0.8976	0.1731	0.3942	0.2331	0.3533	0.4495	0.2590	0.2419	9.1774	0.2762	16	22	14	46	4.2
Min	0.6157	0.5280	0.1352	0.1717	0.8123	0.1516	0.3428	0.2049	0.3136	0.3858	0.2016	0.2120	7.5	0.2563	14	20	12	44	2.4
Max	0.6683	0.6254	0.1762	0.2280	1.0817	0.1898	0.4360	0.2560	0.4125	0.5613	0.3225	0.2792	10.875	0.3073	18	24	16	52	5.6
SE / SIQ	0.0032	0.0051	0.0025	0.0032	0.014	0.0024	0.0044	0.0027	0.0055	0.0113	0.0076	0.0039	0.196	0.0028	1	0.75	1	2.875	0.242
N	20	21	20	21	21	21	20	21	21	21	21	21	21	19	18	21	21	11	21
Kwando: Nkasa (&)
Mean / median	0.6575	0.6069	0.1621	0.2109	0.8470	0.1807	0.3927	0.229	0.3183	0.4328	0.2364	0.2498	11.0812	0.2972	15	21	14	48	5
Min	0.6411	0.5955	0.1515	0.2049	0.7612	0.1714	0.3428	0.2103	0.2985	0.3882	0.2055	0.2413	8.5455	0.2743	12	21	12	46	3.3
Max	0.6765	0.6250	0.1727	0.2185	0.8884	0.1861	0.4147	0.2394	0.3462	0.4681	0.2649	0.2576	13.3636	0.3245	17	23	16	50	6
SE / SIQ	0.0059	0.0047	0.0036	0.0025	0.0179	0.0022	0.0039	0.0043	0.0059	0.0114	0.0084	0.0032	0.7159	0.0081	0.375	0.75	0	0.5	0.45
N	6	6	6	6	7	6	6	6	7	7	7	6	7	6	7	7	7	6	6
Kwando: Linyanti (L)
Mean / median	0.6513	0.5984	0.1629	0.2134	0.8682	0.1767	0.4023	0.2306	0.3326	0.4501	0.2023	0.2402	8.8352	0.2967	17	23	16	50	5.5

( Continued ) Table 2.1. (Continued).

PDL /	PAL /	LD /	LA /	LPF /	PPF /	pD / SL	CPL /	CPD /	LS(o) /	OD /	HL /	HL / Na	BD /	nD	nA	SPc	SLS	SL (cm)
SL	SL	SL	SL	HL	SL	SL	CPL	HL	HL	SL	SL
Min	0.6280	0.5755	0.1458	0.1849	0.7912	0.1555	0.3257	0.2063	0.2838	0.4085	0.1677	0.2228	7.7222	0.2578	15	20	12	46	3.5
Max	0.6731	0.6295	0.1792	0.2348	0.9643	0.2005	0.4341	0.2565	0.3937	0.4961	0.2379	0.2635	9.5385	0.3277	18	25	16	52	6.5
SE / SIQ	0.0019	0.0021	0.0017	0.0021	0.0095	0.0019	0.0039	0.0027	0.0058	0.0036	0.0033	0.0016	0.086	0.0029	0.5	0.5	1	1.5	0.126
N	29	29	29	29	29	29	29	29	29	29	29	29	29	29	29	29	28	29	29
Okavango (O1,O2)
Mean / median	0.6463	0.5954	0.1664	0.2143	0.8577	0.1474	0.3933	0.2123	0.2993	0.4159	0.2049	0.2407	9.0654	0.2504	16	22	12	49	4.4
Min	0.5995	0.5698	0.1391	0.1864	0.7849	0.1296	0.3402	0.1914	0.2596	0.3761	0.1817	0.2213	8.0	0.2171	14	20	12	46	3.4
Max	0.6864	0.6272	0.1871	0.2366	0.9242	0.1629	0.4143	0.238	0.3521	0.4579	0.23	0.2604	10.4444	0.2895	18	24	14	55	5.4
SE / SIQ	0.0026	0.0021	0.0019	0.0019	0.0067	0.0015	0.0028	0.002	0.0035	0.003	0.0022	0.0015	0.1007	0.0027	0.5	0.875	0.375	1.5	0.092
N	35	35	35	35	35	34	35	35	35	35	35	35	35	35	35	35	35	35	35
P. castelnaui syntypes (BMNH 910.5.31.11-12(2), “Lake Ngami”
Mean / median	0.6474	0.5974	0.1874	0.2373	0.9093	0.1474	0.3903	0.2101	0.3286	0.4187	0.2353	0.2248	10.7692	0.3033	18	23	12	52.5	6.2
Type 1	0.6473	0.5907	0.1947	0.2463	0.8939	0.1296	0.3877	0.1997	0.3333	0.4361	0.2299	0.2213	10.2308	0.2928	18	23	12	52	6
Type 2	0.6475	0.6040	0.1801	0.2283	0.9248	0.1629	0.3929	0.2205	0.3239	0.4014	0.2406	0.2283	11.3077	0.3137	18	23	12	53	6.4
1 / range 2	0.0002	0.0067	0.0073	0.0090	0.0155	0.0167	0.0026	0.0104	0.0047	0.0174	0.0054	0.0035	0.5385	0.0104	0	0	0	0.5	0.215

Notes: SE, standard error. Median and SIQ (semi-interquartiles), for count measures (nD, nA, SPc, SLS). For locality symbols, refer to map of Figure 1. Table 2.2. Morphological measures for samples of the Pollimyrus castelnaui / marianne species complex from various origins, sample collected in 2004 for additional EOD and DNA analyses. For abbreviation of morphological characters, see Material and methods.

PDL / SL	nD	nA	SPc	SL (cm)
Zambezi (Z1, 2004)
Mean / median	0.6416	17	24	16	3.4
Min	0.616	17	23	13	2.6
Max	0.6789	19	25	16	4.6
SE / SIQ	0.0033	0.5	0.5	0	0.089
N	23	23	23	23	23
Kwando: Kongola (K, 2004)
Mean / median	0.6556	16.5	22	14	3.1
Min	0.6374	15	21	12	2.3
Max	0.6971	17	23	16	3.9
SE / SIQ	0.0039	0.5	0.5	0.25	0.101
N	20	20	20	20	20
Okavango (O1, 2004)∗
Mean / median	0.6446	17	23	12	3.1
Min	0.6262	16	21	12	2.6
Max	0.6651	18	24	14	3.8
SE / SIQ	0.0046	0	0.5	1	0.139
N	9	9	9	9	9

Notes: SE, standard error. Median and SIQ (semi-interquartiles), for count measures (nD, nA, SPc). For localities Z1, K, O1 refer to map of Figure 1. ∗ After EOD recording, four additional specimens were too damaged to take tissue samples, but were good for DNA. Table 2.3. Principal component analysis on correlations for morphological characters of the Pollimyrus castelnaui / marianne species complex. For abbreviations of morphological characters, see Material and methods.

Eigenvalue	5.4323	2.4326	1.5939	1.2067	1.1324	0.8608	0.7524
Percent	31.954	14.31	9.376	7.098	6.661	5.064	4.426
Cum %	31.954	46.264	55.64	62.738	69.4	74.463	78.889
Component loadings
PDL / SL	−0.7401	0.0695	0.239	0.205	−0.2048	−0.0802	0.159
PAL / SL	−0.8328	0.0845	0.2058	0.071	−0.0814	−0.0361	0.0797
LD / SL	0.6561	−0.0058	0.3642	0.2598	0.1459	0.0144	−0.3464
LA / SL	0.7108	0.0639	0.3863	0.1587	−0.1981	0.0909	0.0092
LPF / HL	0.5304	0.248	0.0095	−0.0916	−0.4416	0.0445	0.0507
PPF / SL	−0.537	0.6505	−0.0172	0.2935	−0.0568	−0.1298	−0.0338
pD / SL	0.6038	0.2217	−0.2482	0.2319	0.3631	0.1691	−0.2982
CPL / SL	0.1693	0.4756	−0.7559	0.1269	0.118	−0.1877	0.0798
CPD / CPL	−0.3864	0.4514	0.4795	−0.0915	0.1863	0.5012	−0.0687
LSo / HL	−0.2065	0.5602	−0.2375	−0.4011	−0.4207	0.0559	−0.2725
OD / HL	−0.4888	0.2267	−0.0656	−0.5734	0.176	0.2088	−0.0293
HL / SL	−0.7224	−0.0945	−0.0074	0.0942	0.5078	−0.0326	0.1967
BD / SL	−0.3604	0.6788	0.2062	0.4038	−0.0915	−0.1777	−0.0761
nD	0.4029	0.3549	0.299	−0.3322	0.3169	−0.4628	−0.1522
nA	0.4849	0.3218	0.3764	−0.3362	0.1298	−0.3201	0.2876
SPc	0.4592	0.6147	−0.1183	0.0666	0.201	0.3194	0.3206
SLS	0.7756	0.1025	0.0294	0.0692	−0.1129	0.113	0.4107

Notes: N = 155 specimens from Okavango ( N = 34), Upper Zambezi ( N = 61), and Kwando : Kongola Bridge ( N = 15), Nakatwa ( N = 11), Nkasa ( N = 6), Linyanti / Sampis ( N = 28) . Table 2.4. ISSR profiles of P. castelnaui , P. marianne and P. cuandoensis sp. nov. Shown are the informative bands nos 1 to 13; X = band is present. X symbols of bold and underlined style (in colour online) are highlighting bands characteristic of species.

Taxon	IPMB	1	2	3	4, 10, 11,	5	6	7	8	9
ID	12, 13
castelnaui
16555	X	X	X	X	X
16558	X	X	X	X	X
16561	X	X	X	X	X
44698	X	X	X	X
45377	X	X	X	X	X
45378	X	X	X	X	X	X
marianne
44647	X	X	X	X
44648	X	X	X	X	X
44651	X	X	X	X	X
44652	X	X	X	X
44654	X	X	X	X
44655	X	X	X	X
44656	X	X	X	X	X	X
44657	X	X	X	X	X	X
44658	X	X	X	X	X
44659	X	X	X	X	X
44660	X	X	X	X	X	X
44661	X	X	X	X	X	X
44662	X	X	X	X	X
44663	X	X	X	X	X	X
44664	X	X	X
44665	X	X	X	X	X
44666	X	X	X	X	X
44667	X	X	X	X
44668	X	X	X	X	X
cuandoensis
sp. nov.
44669	X	X	X	X
44670	X	X	X	X	X	X
44671	X	X	X	X	X	X
44672	X	X	X	X	X	X
44673	X	X	X	X	X	X
44674	X	X	X	X
44675	X	X	X	X
44676	X	X	X	X	X	X
44677	X	X	X	X	X	X	X
44679	X	X	X	X	X	X	X	X
44682	X	X	X	X	X	X

Appendix 3. Electrical characters for different populations of P.cuandoensis sp. nov. from the Kwando River Table 3.1. EOD waveform characters for local populations of Pollimyrus cuandoensis sp. nov. in the Kwando River. For abbreviations, see Material and methods.

Sample	P0 (V)	N0 (V)1	P1 (V)	P2 (V)	P0dur	N0dur	P1dur	N1dur	P2dur	EODdur	P0N0sep	N0P1sep	P1N1sep	P0area	N0area	P1area	N1area	P2area	SL (cm)
origin	(ms)	(ms)	(ms)	(ms)	(ms)	(ms)	(ms)	(ms)	(ms)	( V × ms)	( V × ms)	( V × ms)	( V × ms)	( V × ms)
Kongola (K)
Mean	0.1273	−1.404	0.2735	0.0957	0.0656	0.066	0.0608	0.0747	0.2374	0.5045	0.0645	0.0782	0.0443	0.062	0.0061	0.0099	0.0395	0.0178	4.37
SE	0.0076	0.0064	0.0047	0.0027	0.0027	0.0018	0.0013	0.0009	0.0077	0.0083	0.0011	0.0016	0.005	0.005	0.0004	0.0003	0.0005	0.0009	0.127
Min	0.0209	−0.0212	0.1495	0.0522	0.0201	0.0223	0.0366	0.0594	0.0859	0.3435	0.0421	0.0576	0.0359	0.0003	0.0003	0.0045	0.0295	0.0037	2.45
Max	0.2827	−0.2756	0.3385	0.1614	0.1127	0.1086	0.0836	0.0939	0.3582	0.6357	0.0926	0.1107	0.054	0.0181	0.015	0.0139	0.0518	0.034	6.1
N	63	63	63	63	63	63	63	63	63	63	63	63	63	63	63	63	63	63	63
Nakatwa (N)
Mean	0.1321	−0.1116	0.2683	0.086	0.0568	0.0496	0.0482	0.063	0.1914	0.4090	0.0481	0.063	0.0362	0.0055	0.0037	0.0073	0.0333	0.0131	4.37
SE	0.0122	0.0088	0.009	0.0024	0.0035	0.0019	0.0014	0.0014	0.0085	0.117	0.0018	0.0012	0.0005	0.0007	0.0004	0.0004	0.0007	0.0007	0.232
Min	0.0506	−0.0519	0.1707	0.0571	0.0208	0.0363	0.0353	0.0527	0.1067	0.2976	0.0315	0.056	0.0324	0.0008	0.0012	0.0035	0.0273	0.0048	2.42
Max	0.2934	−0.2052	0.3407	0.1097	0.0874	0.0703	0.0643	0.0816	0.2648	0.5156	0.064	0.0823	0.0412	0.0149	0.008	0.0098	0.0393	0.019	5.58
N	24	24	24	24	24	24	24	24	24	24	24	24	24	24	24	24	24	24	24
Nkasa (&)
Mean	0.1015	−0.113	0.2527	0.0992	0.0568	0.0651	0.06	0.0769	0.2394	0.4982	0.0654	0.0736	0.0465	0.0046	0.0047	0.0089	0.0416	0.0184	4.96
SE	0.0218	0.0111	0.013	0.0086	0.0092	0.0048	0.005	0.0027	0.0294	0.0349	0.0042	0.0039	0.0027	0.0014	0.0008	0.0009	0.0015	0.0034	0.39
Min	0.0267	−0.0647	0.2133	0.0711	0.0224	0.0519	0.0466	0.0691	0.1309	0.3989	0.0511	0.0596	0.0406	0.0004	0.0022	0.0059	0.0369	0.0075	3.45
Max	0.2008	−0.1445	0.3001	0.1424	0.0841	0.0897	0.0797	0.0865	0.3781	0.6571	0.0827	0.09	0.0585	0.0104	0.0081	0.0131	0.0466	0.0368	6
N	7	7	7	7	7	7	7	7	7	7	7	7	7	7	7	7	7	7	7
Linyanti (L)
Mean	0.1019	−0.1023	0.2846	0.1028	0.0479	0.0518	0.0543	0.0757	0.2507	0.4804	0.0558	0.0652	0.0417	0.0045	0.0036	0.009	0.0407	0.0193	5.72
SE	0.0164	0.0097	0.0123	0.0033	0.0049	0.0021	0.0024	0.0013	0.0076	0.0083	0.0019	0.002	0.005	0.0011	0.0004	0.0006	0.0005	0.0009	0.109
Min	0.0147	−0.0089	0.1072	0.0523	0.0173	0.022	0.0257	0.0658	0.1382	0.4097	0.035	0.0405	0.0349	0.0002	0.0001	0.0018	0.036	0.0058	3.53
Max	0.3733	−0.225	0.4107	0.1457	0.1184	0.076	0.0908	0.0981	0.3522	0.6338	0.0736	0.0833	0.05	0.0223	0.0099	0.0146	0.0474	0.0311	6.71
N	33	33	33	33	33	33	33	33	33	33	33	33	33	33	33	33	33	33	33

1 Negative values for N0: maximum and minimum reversed (that is, taken absolute). For locality symbols in parentheses, refer to map of Figure 1. Table 3.2. Principal component analysis on correlations for 17 EOD characters of specimens of Pollimyrus cuandoensis sp. nov. from various origins of the Kwando / Linyanti system. For abbreviations, see Material and methods.

Eigenvalue	6.2355	4.8092	2.2914	1.4928	0.7926	0.4408	0.3903
Percent	36.68	28.29	13.479	8.781	4.662	2.593	2.296
Cum Percent	36.68	64.969	78.448	87.229	91.892	94.485	96.781
Component loadings
P0 (V)	−0.714	0.6126	0.1742	0.116	0.2115	−0.0815	0.0325
N 0 ( V )	0.0681	−0.8348	0.3909	0.2037	−0.2166	0.1045	−0.1358
P1 (V)	0.6023	−0.4413	0.0008	−0.1172	0.5322	0.3364	0.1626
P2 (V)	0.8293	0.0634	−0.4025	0.2016	−0.1325	−0.0526	0.074
P0dur (ms)	−0.6256	0.5879	0.3698	−0.0142	0.0953	−0.0247	0.1228
N 0dur (ms)	0.4455	0.7028	−0.2175	−0.4378	−0.069	0.0843	−0.1182
P1dur (ms)	0.7253	−0.2072	0.5629	−0.225	−0.027	−0.2089	−0.0166
N 1dur (ms)	0.2181	0.6483	0.5036	0.448	0.0538	0.1232	−0.1536
P2dur (ms)	0.6771	0.4022	−0.3887	0.389	0.0738	−0.0519	−0.0597
P0N0sep	0.1621	0.6993	0.1419	−0.3913	−0.3639	0.3789	−0.0857
(ms)
N0P1sep	0.7236	0.2032	0.3582	−0.4045	0.1029	−0.2808	−0.2016
(ms)
P1N1sep	0.5707	0.3267	0.4606	0.1408	−0.3453	−0.0398	0.4442
(ms)
P0area	−0.6841	0.6286	0.2901	0.1048	0.1482	−0.0167	0.0088
( V × ms)
N0area	0.176	0.8298	−0.3626	−0.3016	0.144	−0.0663	0.1129
( V × ms)
P1area	0.8076	−0.2415	0.4032	−0.2162	0.2421	0.0365	0.083
( V × ms)
N1area	0.6485	0.4429	0.3483	0.4553	0.1165	0.0792	−0.1322
( V × ms)
P2area	0.7755	0.314	−0.4252	0.3103	−0.013	−0.0433	−0.0003
( V × ms)

Notes: N = 127 specimens : Kongola Bridge ( N = 63), Nakatwa ( N = 24), Nkasa ( N = 7), Linyanti / Sampis ( N = 33).