3.4. Phylogenetic placement of
Paranybelinia otobothrioides
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The 1989 bp concatenated fragment of the small subunit rDNA (ssrDNA) of
Pa. otobothrioides
showed the nucleotide composition proportion of T = 25.4, C = 22, A = 23.9, G = 28.7, and G + C = 50.7%. The analyzed molecular data set consisted of 58 trypanorhynch sequences, including 11 species belonging to the
Tentaculariidae
and 2 species belonging to the
Rhinoptericolidae
, both families considered to be related with the
Paranybeliniidae
. The aligned 18S partition comprised 1913 positions of which 525 (26%) positions were variable and 400 (20%) were parsimony informative.
The Maximum likelihood analysis of the ssrDNA was largely congruent with the known topology of trypanorhynch cestodes (see Palm et al., 2009; Olson et al., 2010) with two major clades, the suborders Trypanoselachoida and
Trypanobatoida
(bootstrap values range 70–100). Inside the suborder
Trypanobatoida
, the genera subtropical. krill
Nyctiphanes simplex
. SD = Standard deviation.
Prochristianella
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,
Parachristianella
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,
Trimacracanthus
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(Eutetrathynchiidae) together with
Trygonicola
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and
Halysiorhynchus
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(
Mixodigmatidae
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) formed a high supported clade (bootstrap 100%). This group placed sister to a weak supported clade (bootstrap <70%), consisting of
H. australiensis
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,
Mecistobothrium johnstonei (Southwell, 1929) Beveridge & Campbell, 1998
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and
Oncomegas celatus (Beveridge and Campbell, 2005) Schaeffner and Beveridge, 2012
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[=
Oncomegoides celatus
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]) and the well supported clade formed by the
Tetrarhynchobothrium Diesing, 1854
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,
Dollfusiella Campbell and Beveridge, 1994
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, and
Paroncomegas Campbell, Marques and Ivanov, 1999
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(bootstrap 100%) ( Fig. 5
View Fig
).
The superfamily
Tentacularioidea
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was represented in a distinct major clade (bootstrap 100%) ( Fig. 5
View Fig
). The phylogenetic analysis placed
Pa. otobothrioides
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in a clade with high nodal support (bootstrap 100%) as sister taxon to the tentaculariids. The species
Rhinoptericola megacantha Carvajal and Campbell, 1975
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and
Nataliella marcelli Palm, 2010
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(
Rhinoptericolidae
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), despite considered as members of the
Eutetrarhynchoidea
, were placed as sister taxon to the superfamily
Tentacularioidea
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(i.e.,
Tentaculariidae
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and
Paranybeliniidae
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) with strong nodal support (bootstrap> 90).
Estimates of evolutionary divergence based on Kimura-2-parameters corrected distances (K2P) and the number of base differences per site between sequences were calculated between the
Tentacularioidea
( Table 2), including the
Tentaculariidae
and the
Paranybeliniidae
.
Paranybelinia otobothrioides
showed no intraspecific genetic variation. Estimates of K2P between
Pa. otobothrioides
and the 11 tentaculariid species ranged from 0.027 to 0.039 (average = 0.033) and the number of base differences per sequence (nd) ranged from 44 to 62. Inside the
Tentaculariidae
, the genetic distance resulted in a considerably lower K2P range from 0.003 to 0.03 (average = 0.018) with the nd ranging from 5 to 49. Of the analyzed
Tentaculariidae
,
Tentacularia coryphaenae Bosc, 1802
was the closest species to
Pa. otobothrioides
(K2P = 0.027, nd = 44), whilst
Nybelinia africana Dollfus, 1960
showed to be the most divergent sequence (K2P = 0.039, nd = 62) ( Table 2).