Research Article |
Corresponding author: Ioana Cristina Constantinescu ( cristinactinescu@yahoo.com ) Academic editor: Vladimir Pesic
© 2018 Ioana Cristina Constantinescu, Oana Paula Popa, Luis Ovidiu Popa, Ioana Cobzaru, Mukhim D. Khlur B., Costică Adam.
This is an open access article distributed under the terms of the Creative Commons Attribution License (CC BY 4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Citation:
Constantinescu IC, Popa OP, Popa LO, Cobzaru I, Khlur B MD, Adam C (2018) A new feather mite species of the genus Trouessartia Canestrini, 1899 (Acarina, Trouessartiidae) – an integrative description (morphology and DNA barcoding data). ZooKeys 789: 19-35. https://doi.org/10.3897/zookeys.789.27829
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A new species of the feather mite genus Trouessartia (Trouessartiidae) is described from the Large NiltavaNiltava grandis (Blyth) (Passeriformes, Muscicapidae) in Northeast India (Meghalaya, Jaintia Hills, Shnongrim village). Trouessartia niltavae Constantinescu, sp. n. is morphologically closely related (no phylogenetic meaning) to T. bulligera Gaud, 1968 from Clytorhynchus hamlini (Mayr) (Passeriformes: Monarchidae), sharing in males a unique character within the genus, by having setae e on legs IV hemispheroid, with spine-shaped apex. Males of the new species have the prodorsal shield without ornamentation, the prohysteronotal shield and lobar shield connected, and the terminal cleft parallel sided. Females have the posterior half of the hysteronotal shield ornamented with large ovate lacunae in central area and small elliptical lacunae marginally. To the morphological description of this new feather mite species we added sequence data on the mitochondrial cytochrome c oxidase subunit I gene fragment (COI). The phylogenetic relationships between Trouessartia species are briefly discussed.
Feather mite, new species, taxonomy, Trouessartia
The feather mite genus Trouessartia Canestrini, 1899 comprises about 120 described species associated predominantly with birds of the order Passeriformes. A world revision of this genus, including 71 species was performed by
The bird genus Niltava Hodgson belongs to the family of Old World Flycatchers (Passeriformes: Muscicapidae) and currently includes six valid species distributed in the Indo-Malayan biogeographic region (
The main goals of this paper are to realise the description of a new species of Trouessartia and to analyse its relationships within the genus based on molecular data. This is the first species of Trouessartia described from a host of the genus Niltava, although, as mentioned above, two presumably new species of Trouessartia have been reported by Atyeo (
The new species of Trouessartia described herein cannot be referred to any of the seven species groups previously established in the genus (
The material used in the present paper was collected near Shnongrim (Meghalaya, India) in January 2014. The birds were captured using mist nets, identified and visually checked for the presence and collection of mites and released back to the wild. Mite specimens were collected manually with a needle and placed in tubes with 96% ethanol. Later, in laboratory conditions, mite specimens selected for morphological analysis were cleared in 90% lactic acid for 24 hours and mounted on microscope slides in Hoyer’s medium. Some specimens preserved in ethanol were used for genetic analysis. Drawings were made using an Olympus CX21 microscope, with a camera lucida drawing device. The bird specimens were identified according to
Three paratype specimens of Trouessartia niltavae sp. n. (one male ANA838 and two females ANA839, ANA840) were used to isolate DNA using DNAeasy Tissue Kit (Qiagen). All four specimens used for molecular analyses were mounted and kept as reference vouchers for morphological examination. The specimens preserved in ethanol 96% were transferred in 180 μl ATL Buffer with 20 μl of Proteinase K and incubated overnight at 56 °C on a shaking thermoblock. After 24 h, 5μl of Proteinase K were added and incubation was continued until 72 h. For the rest of the protocol we followed the manufacturer specifications and the modification suggested by
As DNA barcode we used a region near the 5` terminus of the COI gene, amplified by PCR with the degenerate primers bcdF05 (5`- TTTTCTACHAAYCATAAAGATATTGC-3`) and bcdR04 (5`- TATAAACYTCDGGATGNCCAAAAAA-3`), according to
Sequence chromatograms were edited and assembled with CodonCode Aligner version 3.7.1. For the phylogenetic analysis we used a dataset comprising three sequences obtained from the new species Trouessartia niltavae and 74 sequences belonging to 17 species of Trouessartia genus available in GenBank and BOLD data system and four sequences belonging to genus Calcealges (see Tabel 1), to be used as outgroup.
MEGA version 7 software (
Male holotype (ANA663), 5 male (ANA661, ANA662, ANA664, ANA665, ANA838) and 8 female (ANA666, ANA667, ANA668, ANA669, ANA670, ANA839, ANA840) paratypes, 27.01.2014, from the Large NiltavaNiltava grandis grandis (Blyth) (Passeriformes, Muscicapidae); INDIA: Meghalaya, Jaintia Hills, Shnongrim village, (25°21’12.36”N, 92°31’3.06”E); 1151 m; subtropical forest; collector D. K. B. Mukhim.
Male (Figs
Epimerites I free. Rudimentary sclerites rEpIIa present, roughly rounded. Genital apparatus situated between levels of trochanters III and IV, length excluding basal sclerite 38 (37–40), greatest width 14 (10–14) (Fig.
Female (Figs
Epimerites I free. Epigynum 36–38 in length, 76–80 in width (Fig.
The specific name niltavae is derived from the generic name of the type host and is a noun in the genitive case.
Trouessartia niltavae sp. n. is morphologically close to T. bulligera Gaud, 1968 from Clytorhynchus hamlini (Mayr) (Passeriformes: Monarchidae), sharing in males a unique character within the genus: setae e on tarsi IV are hemispheroid with stick-like apex. Additionally, in both sexes of these species, setae d1 are present, setae c2 and sRIII are narrowly lanceolate, with acute apex and the dorsal hysterosomal apertures (DHA) is absent. Both sexes of T. niltavae differ from those of T. bulligera by the shape of setae c2, which are spiculiform in the first species versus needle-like in the second. Males of both species have the prohysteronotal shield without ornamentation, the lamellae of opisthosomal lobes are attenuate apically and with entire margins, the translobar apodeme is present, setae g are contiguous at bases and situated on postgenital plaque. In males of T. niltavae, the prodorsal shield is without ornamentation, the prohysteronotal shield and lobar shield have wide median connection, the terminal cleft is parallel-sided and 44-46 μm long, and terminal lobes are separated by 6–8 μm. In males of T. bulligera, the prodorsal shield has ornamentation with faint, interconnecting network of irregular lines, the prohysteronotal shield is completely separated from the lobar shield, the terminal cleft is divergent in posterior half and 75 μm in length, and terminal lobes are separated by 12 μm. In females of both species, setae h1 are lanceolate, the external copulatory tube is present, the supranal concavity is open posteriorly into terminal cleft, and the interlobar membrane occupies the anterior 1/3 of terminal cleft. In females of T. niltavae setae f2 are present, the posterior half of the hysteronotal shield is ornamented with large ovate lacunae in the central area and small elliptical lacunae marginally arranged. In females of T. bulligera setae f2 are absent, the posterior half of the hysteronotal shield has ornamentation with small elliptical lacunae in the central area and large ovate lacunae marginally arranged.
DNA barcode. Representative DNA sequences: molecular voucher specimens ANA838 male (GenBank accession number MH094247), ANA839 female (GenBank accession number MH094248), ANA840 female (GenBank accession number MH094249).
We sequenced a 586-pb fragment of the mitochondrial cytochrome c oxidase subunit I (COI) gene for two females and one male paratypes. All three sequences belong to a single haplotype. The calculated intra-specific genetic distances (K2P) for other species of Trouessartia was as follows: Trouessartia rosterii 0,8%, T. reguli 1,4%, T. kratochvili 1%, T. rubecula 0,5%, T. simillima 0.4%, T. ripariae 0,4%, T. microcaudata 0,7%, T. tenuipillata 0,6%, T. jedliczkai 1,6%, T. trouessarti 1.4%, T. motacillae 1.1%, T. bifurcata 11,5%.
The best-fit base substitution model for the analyzed data was determined to be TN93+G+I. The NJ and ML trees exhibited similar topologies and branching structures (see Figures
Analysed sequences of Trouessartia spp. from BOLD and GenBank databases.
Species | GenBank Accession Number | Species | GenBank Accession Number | Species | GenBank Accession Number |
T. appendiculata | KP193765 | T. rubecula | KP193799 | T. rosterii | KP193796 |
T. bifurcata | KP193766 | T. rubecula | KP193801 | T. rosterii | KP193797 |
T. bifurcata | KP193767 | T. rubecula | KP193802 | T. rosterii | KP193798 |
T. inexpectata | KP193768 | T. rubecula | KP193803 | T. reguli | MG411516 |
T. jedliczkai | KP193769 | T. rubecula | KP193804 | T. reguli | MG409826 |
T. jedliczkai | KP193770 | T. rubecula | KP193805 | T. reguli | MG409631 |
T. jedliczkai | KP193771 | T. rubecula | KP193806 | T. reguli | MG414726 |
T. jedliczkai | KP193772 | T. rubecula | KP193807 | T. reguli | MG413144 |
T. jedliczkai | KP193773 | T. rubecula | KP193808 | T. reguli | MG410216 |
T. jedliczkai | KP193774 | T. rubecula | KP193809 | T. reguli | MG412618 |
T. kratochvili | KP193776 | T. rubecula | KU203092 | T. reguli | MG411130 |
T. kratochvili | KP193777 | T. simillima | KP193810 | T. reguli | MG416767 |
T. kratochvili | KP193778 | T. simillima | KP193811 | T. reguli | MG414272 |
T. kratochvili | KU203094 | T. simillima | KP193812 | T. reguli | KP193788 |
T. microcaudata | KP193779 | T. swidwiensis | KP193813 | T. reguli | KP193789 |
T. microcaudata | KP193780 | T. tenuipilata | KP193814 | T. reguli | KP193790 |
T. microcaudata | KP193781 | T. tenuipilata | KP193815 | T. reguli | KP193791 |
T. microcaudata | KP193782 | T. tenuipilata | KP193816 | T. reguli | KU203095 |
T. minuscula | KP193783 | T. trouessarti | KP193817 | T. reguli | KU203096 |
T. motacillae | KP193784 | T. trouessarti | KP193818 | T. niltavae sp. n. | MH094247 * |
T. motacillae | KP193785 | T. trouessarti | KP193819 | T. niltavae sp. n. | MH094248 * |
T. motacillae | KP193786 | T. rosterii | KT025283 | T. niltavae sp. n. | MH094249 * |
T. motacillae | KP193787 | T. rosterii | KT025284 | Calcealges sp. | MG410916 |
T. ripariae | KP193792 | T. rosterii | KT025288 | Calcealges sp. | MG412689 |
T. ripariae | KP193793 | T. rosterii | KT025289 | Calcealges sp. | KU203091 |
T. ripariae | KP193794 | T. rosterii | KP193795 | Calcealges sp. | MG409226 |
We are grateful to the Additional Principal Chief Conservator of Forests, Wildlife & Chief Wildlife Warden from Shillong (Meghalaya, India) for the permission to catch birds (permission No. FWC.G/173/Pt.). Ioana Cobzaru was supported by the Institute of Biology Bucharest of Romanian Academy (project no. RO1567–IBB04/2016).