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Volleth, Marianne, Son, Nguyen Truong, Wu, Yi, Li, Yuchun, Yu, Wenhua, Lin, Liang-Kong, Arai, Satoru, Trifonov, Vladimir, Liehr, Thomas, Harada, Masashi (2017): Comparative chromosomal studies in Rhinolophus formosae and R. luctus from China and Vietnam: elevation of R. l. lanosus to species rank. Acta Chiropterologica 19 (1): 41-50, DOI: 10.3161/15081109ACC2017.19.1.003

Rhinolophus formosae Two males and one female from Taiwan were studied cytogenetically. All specimens showed a karyotype with 2 n = 52 and a FNa = 60 (Fig. 6). There were five bi-armed and 19 acrocentric autosomal pairs. In addition, the smallest, dot-like pair no. 25 is likely also bi-armed. Due to the minuteness of this element, the metacentric condition was visible only in a small percentage of metaphase spreads. Therefore, this chromosome was counted as one arm only for the FNa. The medium-sized X chromosome is submetacentric and the dot-like Y chromosome is probably bi-armed. Concerning the amount of centromeric heterochromatin, the acrocentric pairs differed from the bi-armed ones. In contrast to the faint staining of centromeric regions of bi-armed pairs 1 to 5 and 25, the acrocentric chromosomes showed dark stained pericentromeric regions after CBG-banding, which were also GTG-, QFQ- and DAPI-positive (Fig. 7 A���B). The amount of centromeric heterochromatin of the X chromosome was similar to that of other Rhinolophus species and not enlarged as in R. cf. luctoides and R. lanosus. The Y chromosome of R. formosae was hardly distinguishable from the autosomal pair 25 after CBG-banding and showed no clear centromeric heterochromatin (Fig. 7A). The secondary constriction of chromosomal pair 18 (homologous to MMY21) was shown to bear active NORs by silver-staining (Fig. 7C). Analysis of 20 metaphase spreads of one male specimen revealed a frequency of 2.0 NORs per cell. The complete set of AST painting probes and some selected MMY probes (MMY8, 14, 23) were applied on R. formosae. The results are given on the G-banded karyogram (Fig. 6) and examples of FISH experiments are shown in Fig. 3C and 3F. Of the chromosomal pairs 1 to 5, only two, i.e., 4 and 5, show the same combination of chromosomal arms as found in R. luctoides, R. lanosus and R. morio. Pairs 1 to 3 show a unique combination hitherto found in no other rhinolophid species., Published as part of Volleth, Marianne, Son, Nguyen Truong, Wu, Yi, Li, Yuchun, Yu, Wenhua, Lin, Liang-Kong, Arai, Satoru, Trifonov, Vladimir, Liehr, Thomas & Harada, Masashi, 2017, Comparative chromosomal studies in Rhinolophus formosae and R. luctus from China and Vietnam: elevation of R. l. lanosus to species rank, pp. 41-50 in Acta Chiropterologica 19 (1) on pages 44-45, DOI: 10.3161/15081109ACC2017.19.1.003, http://zenodo.org/record/3944816

Rhinolophus cf. luctoides The male from Vietnam, morphologically determined as R. luctus, showed a karyotype with a diploid chromosome number of 2 n = 32 and an autosomal fundamental number of FNa = 60. The karyotype consisted of 15 meta- to submetacentric autosomal pairs ranging from large to small, a large metacentric X chromosome and a large acrocentric Y chromosome (Fig. 1). C-banding revealed heterochromatin at all centromeres, in the proximal half of the short arm of the X chromosome, and in the Y chromosome with exception to the very proximal part (Fig. 2A). The secondary constriction in the short arm of chromosomal pair 8 (homologous to MMY21 and AST10) was proven to be the single site of Nucleolus Organizer Region (NOR) by silver-staining (Fig. 2B). The mean number of active NORs per cell was 1.7 (20 cells analyzed). The G-banding pattern of autosomal pairs was found to be similar to R. luctoides (Volleth et al., 2015). The identical arm composition of autosomes as in R. luctoides was confirmed by the results of whole chromosome painting with probes from A. stoliczkanus (AST), complemented with some probes from M. myotis (MMY3/4 and MMY8 for pair 1). The hybridization results are summarized on the karyotype (Fig. 1). Painting probes from biarmed AST autosomes hybridized to two different pairs each (examples in Fig. 3A and 3D)., Published as part of Volleth, Marianne, Son, Nguyen Truong, Wu, Yi, Li, Yuchun, Yu, Wenhua, Lin, Liang-Kong, Arai, Satoru, Trifonov, Vladimir, Liehr, Thomas & Harada, Masashi, 2017, Comparative chromosomal studies in Rhinolophus formosae and R. luctus from China and Vietnam: elevation of R. l. lanosus to species rank, pp. 41-50 in Acta Chiropterologica 19 (1) on page 43, DOI: 10.3161/15081109ACC2017.19.1.003, http://zenodo.org/record/3944816, {"references":["VOLLETH, M., J. LOIDL, F. MAYER, H. - S. YONG, S. MULLER, and K. - G. HELLER. 2015. Surprising genetic diversity in Rhinolophus luctus (Chiroptera: Rhinolophidae) from Peninsular Malaysia: description of a new species based on genetic and morphological characters. Acta Chiropterologica, 17: 1 - 20."]}

Rhinolophus lanosus stat. rev. The female from Sichuan province, China, showed a diploid chromosome number of 2 n = 32 and a FNa = 60. The karyotype was composed of 15 meta- to submetacentric autosomal pairs and two metacentric X chromosomes (Fig. 4). Heterochromatin was detected by C-banding at all centromeres and in the proximal part of the short arm of the X chromosome. The size of this heterochromatic segment differed between both homologs of the female studied (inset of Fig. 4). The NORs are located at the secondary constriction in the short arm of pair 8. This chromosomal arm is homologous to MMY21. The frequency of active NORs as detected by silver-staining was 1.8 NORs per cell (10 metaphase spreads analyzed). The composition of autosomal arms was studied by combination of whole chromosome painting probes from A. stoliczkanus, complemented by some M. myotis probes (MMY8, 14, 18). The hybridization results are summarized on the karyotype (Fig. 4), and as examples metaphase spreads hybridized with probes from AST 1, 2 and 8 are shown (Fig. 3B and 3E). G-banding and FISH results revealed that the karyotype of R. lanosus differs from that of R. luctoides and R. cf. luctoides from Vietnam by a whole arm reciprocal translocation (WART) between pairs 3 and 4. In the karyotype of R. cf. luctoides, pair 3 is composed of arms homologous to AST2 ad AST5, whereas in R. lanosus it consists of arms homologous to AST7 and AST5. Homology to AST1 and AST7 is found in pair 4 of R. cf. luctoides, but to AST1 and AST 2 in R. lanosus (Fig. 5)., Published as part of Volleth, Marianne, Son, Nguyen Truong, Wu, Yi, Li, Yuchun, Yu, Wenhua, Lin, Liang-Kong, Arai, Satoru, Trifonov, Vladimir, Liehr, Thomas & Harada, Masashi, 2017, Comparative chromosomal studies in Rhinolophus formosae and R. luctus from China and Vietnam: elevation of R. l. lanosus to species rank, pp. 41-50 in Acta Chiropterologica 19 (1) on page 44, DOI: 10.3161/15081109ACC2017.19.1.003, http://zenodo.org/record/3944816

In Europe, 2 different diploid chromosome numbers, 2n = 54 and 2n = 56, have been described in the lesser horseshoe bat (Rhinolophushipposideros). The eastern form with 2n = 56 extends from the Czech Republic to Greece. To date, specimens with 54 chromosomes have been reported only from Spain and Germany. This study expands the distributional area of the western variant to Ireland. Strikingly, this distribution of European chromosomal variants is in contrast to the available molecular data that indicate little genetic differentiation of R. hipposideros populations spanning Northwestern to Central Europe. Further, we have developed an optimized protocol for establishing fibroblast cell cultures, suitable for karyotype analyses, from 3-mm wing membrane biopsies. This is a useful technique for cytogenetic studies of endangered bat species, as this non-lethal sampling method imposes only minimum stress to the animal without lasting adverse effects and is routinely used to sample tissue probes for molecular genetic studies in bats. European Research Council University College Dublin

Molecular phylogram depicting the relationships among genera of bats in the subfamily Phyllostominae (highlighted in bold) and the divergence time estimates (node ages) from BEAST analysis of all genes. Nodes with posterior probability values higher than 0.98 in (a) are marked with an *. Abbreviations for node names are as follows: Phyllostominae + Glossophaginae ancestor (PGA), Phyllostominae ancestor (PA), Glossophaginae ancestor (GA); Phyllostomini ancestor (PiA); Lophostoma + Mimon ancestor (L/M); genus Tonatia (T); genus Anoura (A). (PDF 1529 kb)

Detailed calculations of unique fusions and fissions for ACU and TSA and chromosomal associations shared between the analyzed species. (DOCX 29 kb)

Hexacentrus karnyi Griffini, 1909 [A] CH 4982 ♂ The song of this species (two males recorded in the field at night) was in its basic structure (Fig. 15) similar to that of the Asiatic species H. unicolor Serville, 1831 (Heller, 1986) and other Hexacentrus species (e.g., Ichikawa et al. 2006), but it is much slower in echeme and syllable repetition rate. It consisted of a series of echemes, repeated at intervals of 8���12 s (0.5 s in H. unicolor). The syllable repetition rate was around 50 Hz (T= 21 ��C), again much slower than H. unicolor, which heats up before singing and reaches more than 300 Hz. Each echeme (duration about 0.7 s) started with a distinct impulse, probably indicating the opening of the tegmina. Similar isolated impulses can be seen in the song of the other species, but usually shortly after an echeme and produced during a final wing closure. The peak of the spectrum was situated at 9.7 kHz, close to that of 11 kHz observed in H. unicolor (Heller, 1986)., Published as part of Heller, Klaus-Gerhard, Hemp, Claudia, Liu, Chunxiang & Volleth, Marianne, 2014, Taxonomic, bioacoustic and faunistic data on a collection of Tettigonioidea from Eastern Congo (Insecta: Orthoptera), pp. 343-376 in Zootaxa 3785 (3) on pages 368-369, DOI: 10.11646/zootaxa.3785.3.2, http://zenodo.org/record/250508, {"references":["Griffini, A. (1909) Note sopra alcune Phasgonuridae del Congo. Annales de la Societe entomologique de Belgique, 53 (1908), 9 - 28.","Serville, J. G. A. (1831) Revue methodique des insectes de l'ordre des Orthopteres. Annales des Sciences Naturelles Zoologie et Biologie Animale, 22, 28 - 65; 134 - 167; 262 - 292.","Heller, K. - G. (1986) Warm-up and stridulation in the bushcricket, Hexacentrus unicolor Serville (Orthoptera, Conocephalidae, Listroscelidinae). Journal of Experimental Biology, 126, 97 - 109.","Ichikawa, A., Ito, F., Kano, Y., Kawai, M., Tominaga, O. & Murai, T. (Eds.) (2006) Orthoptera of the Japanese Archipelago in color. Sapporo, 688 pp. + 2 CD."]}

Zeuneria Karsch, 1890 Brunner von Wattenwyl (1878) included Zeuneria in Poreuomenini and retained it there (1891) despite Karsch���s idea (1890) to have it in a separate group Zeuneriae. The genus contains four species (see Table 7) and is certainly very closely related to the monotypic Gravenreuthia Karsch, 1892, which is also included in Poreuomenini (Karsch, 1892). 6.6 / 6.15 36,3 / 37 Z. centralis Rehn, 1914 - / 5.2 - / 32 Rehn, 1914, Published as part of Heller, Klaus-Gerhard, Hemp, Claudia, Liu, Chunxiang & Volleth, Marianne, 2014, Taxonomic, bioacoustic and faunistic data on a collection of Tettigonioidea from Eastern Congo (Insecta: Orthoptera), pp. 343-376 in Zootaxa 3785 (3) on page 361, DOI: 10.11646/zootaxa.3785.3.2, http://zenodo.org/record/250508, {"references":["Karsch, F. (1890) Verzeichniss der von Herrn Dr. Paul Preuss auf der Barombi - Station in Deutsch - Westafrika 1890 gesammelten Locustodeen aus den Familien der Phaneropteriden, Mekonemiden und Gryllakriden. Entomologische Nachrichten, Berlin, 16, 353 - 369.","Karsch, F. (1892) Verzeichniss der von Herrn Dr Paul Preuss im Kamerungebirge erbeuteten Orthopteren. Berliner Entomologische Zeitschrift, 37, 65 - 78. http: // dx. doi. org / 10.1002 / mmnd. 18920370112","Rehn, J. A. G. (1914) Orthoptera. I. Mantidae, Phasmidae, Acrididae, Tettigoniidae und Gryllidae aus dem Zentral - Afrikanischen Gebiet, Uganda und dem Ituri - Becken des Kongos. Wissenschaftliche Ergebnisse der deutschen Zentral - Afrika Expedition, 1907 - 1908, Leipzig, 5 (Zool. 3 [1]), 1 - 223."]}

Afromecopoda preussiana (Karsch, 1891) [A] CH 4945, CH 5001 2 ♂♂, CH 5009 ��� 10 2 ♀♀ There are only two published records of A. preussiana outside Cameroon, both in Congo: Lukula (5.24 ��� S, 12.56 ���E) and Umangi (2.21 ��S, 21.4 ��E; Griffini 1908 a). Irangi is quite far from these localities and much more in the East. One undetermined nymph of the genus, however, had been already collected in the Ituri region (Rehn, 1914), north of the Kivu region. With respect to the possibly large variability (Griffini mentions an unusually small male), we consider our specimens as belonging to this species despite the tegmina being considerably shorter than in the types. At the frontal edge, the tegmina show a very narrow, translucent margin similar as described for A. austera by Karsch 1893. * measured from type photos in OSF, Published as part of Heller, Klaus-Gerhard, Hemp, Claudia, Liu, Chunxiang & Volleth, Marianne, 2014, Taxonomic, bioacoustic and faunistic data on a collection of Tettigonioidea from Eastern Congo (Insecta: Orthoptera), pp. 343-376 in Zootaxa 3785 (3) on pages 366-367, DOI: 10.11646/zootaxa.3785.3.2, http://zenodo.org/record/250508, {"references":["Karsch, F. (1891) Uebersicht der von Herrn Dr. Paul Preuss auf der Barombi-Station in Kamerun gesammelten Locustodeen. Anhang: Ueber die Mecopodiden. Berliner Entomologische Zeitschrift, 36, 317 - 346.","Griffini, A. (1908 a) Phasgonuridae africane del R. Museo di Storia Naturale di Bruxelles, 3. Hetrodidae, Mecopodidae, Pseudophyllidae. Memoires de la Societe Entomologique de Belgique, 15, 35 - 64.","Rehn, J. A. G. (1914) Orthoptera. I. Mantidae, Phasmidae, Acrididae, Tettigoniidae und Gryllidae aus dem Zentral - Afrikanischen Gebiet, Uganda und dem Ituri - Becken des Kongos. Wissenschaftliche Ergebnisse der deutschen Zentral - Afrika Expedition, 1907 - 1908, Leipzig, 5 (Zool. 3 [1]), 1 - 223.","Karsch, F. (1893) Locustodeen von Victoria in Kamerun, gesammelt durch Herrn Dr. Paul Preuss. Entomologische Nachrichten, Berlin, 19, 195 - 199."]}

Pardalota asymmetrica Karsch, 1896 [C] CH 4953 ��� 4, CH 4961 ��� 64 5 ♂♂, 1 ♀ The mainly black and white coloured animals (with orange markings on legs and pronotum) have a distinctive threatening display (Fig. 1 B). They open the elevated wings to some extent, go down with the head towards the ground and bulge the brightly orange skin of the neck as it is done also by disturbed Acripeza reticulata (Rentz 2010). Possibly they are imitating a wasp or another Hymenopteran (see the wasp-imitating phaneropterine Aganacris for a similar wing coloration; Nickle 2012). This idea is supported by the black antennae, marked with white rings only in large intervals. However, even the nymphs present orange and black colours (Fig. 1 C), so that an own chemical defence should not be completely excluded. The eggs are flat as typical for Phaneropterinae (Bey-Bienko 1954), and relatively oblong (4.7 mm long, 1.8 mm width, 0.9mm thick; one dark brown egg, in ethanol). Song: In the field and in the laboratory two different types of song could be heard. The short song (about 0.5���2 s; Fig. 6) was heard quite often, while the complicated long song (8���28 s; 4 complete recordings) was presented only in much larger intervals. At the beginning of the short song an animal typically sings microsyllables by opening the tegmina silently and producing few impulses only during a part of the closing movement. After a variable number of these elements, increasing in amplitude, the animal switches to macrosyllables and during the whole closing movement impulses are produced (Fig. 7 A). The syllable repetition rate decreases only very slightly from 23 to 21 Hz (T> 29 ��C). Mostly there are one or a few intermediate syllables in between. At the end of a short song sometimes microsyllabes are added or even another series of micro- and macrosyllables (observed once in a field recording). A long song (Fig. 2) starts with macrosyllables of low amplitude and short duration, but they become quickly longer and louder and remain so for several (up to many) seconds (syllable repetition rate 23 Hz). After some time the animal switches (Fig. 6, 7 B) to another syllable type which looks like a combination of a micro- and a macrosyllable. After a series of impulses one separate impulse (8���10 ms interval) is following. These syllables are repeated for about 2 to 4 s at a repetition rate of 37 Hz. After some intermediate syllables (Fig. 7 C) the final syllable sequence (0.5 to 1 s duration) is enclosed where very loud isolated impulses are produced, similarly as in the microsyllables, but with a much higher intensity, higher movement amplitude and higher repetition rate (55 Hz). The stridulatory file did not show any irregularities., Published as part of Heller, Klaus-Gerhard, Hemp, Claudia, Liu, Chunxiang & Volleth, Marianne, 2014, Taxonomic, bioacoustic and faunistic data on a collection of Tettigonioidea from Eastern Congo (Insecta: Orthoptera), pp. 343-376 in Zootaxa 3785 (3) on pages 347-349, DOI: 10.11646/zootaxa.3785.3.2, http://zenodo.org/record/250508, {"references":["Karsch, F. (1896) Neue Orthopteren aus dem tropischen Afrika. Stettiner Entomologische Zeitung, 57, 242 - 359.","Rentz, D. C. F. (2010) A guide to the katydids of Australia. CSIRO Publishing, Collingwood, 214 pp.","Nickle, D. A. (2012) Synonymies of wasp-mimicking species within the katydid genus Aganacris (Orthoptera: Tettigoniidae: Phaneropterinae). Journal of Orthoptera Research, 21, 245 - 250. http: // dx. doi. org / 10.1665 / 034.021.0209","Bey-Bienko, G. Ya. (1954) Orthoptera Vol. II. No. 2. Tettigonioidea Phaneropterinae, Fauna of the U. S. S. R. Zoological Institute Akademii Nauk SSSR, 381 pp. [English translation 1965 Jerusalem (Israel Program for Scientific Translations)]"]}