Some considerations on the morphological features that differentiate Alectoris barbara barbata by A. b. barbara are exposed and are also reported the results of a genetic investigation performed on historical specimens. Results showed a considerable genetic distance (0.60), certainly enough to consider it an ESU (Evolutionary Significant Unit), but most likely a separate species

Borgo Enrico

Pellegrino Irene

Spanò Silvio

Spanò Silvio

Archivio Istituzionale della Ricerca- Università del Piemonte Orientale

© 2013 CISO - Centro Italiano Studi Ornitologiciprimary, tarsus, tail) on subjects preserved in Museums (7 in Museo Civico di Storia Naturale “G. Doria” of Genoa, 17 in Museo Regionale di Scienze Naturali of Turin). We also analyzed 241 bp from d-loop mtDNA in four A. barbata individuals collected in Cyrenaica (Libya) be-tween 1915 and 1922 and preserved in the Museum of Tu-rin. Total genomic DNA was isolated from feathers with NucleoSpin kit (Macherey-Nagel), according to the man-ufacturer’s instructions with the following modifications: feather tips were placed in a lysis buffer for small DNA quantity (FLB buffer; Macherey-Nagel) and exposed to thermal shock in liquid nitrogen. Sequences of hypervari-able domain I of mitochondrial DNA control region were amplified by polymerase chain reaction using two PCR primers specifically designed. Amplification was carried by the following fast protocol: (94°C x 2’), 30 cycles at (94°C x 30”), (55°C x30”), (72°C x 30”), and a final exten-sion at 72°C for 10 minutes. Amplicons were sequenced at the “BMR genomics” lab (Padova, Italy). Sequence alignment was performed with Bioedit version 7.0.9 (Hall 1999). The Neighbour-joining method (NJ; Saitou & Nei 1987), clustering pairwise Tamura-Nei’s (TN93; Tamura & Nei 1993) genetic distances between haplotypes was performed with MEGA 5.0 (Tamura et al. 2011); support for the internodes in the NJ tree was assessed by bootstrap percentages (BP; Felsenstein 1985) after 1000 resampling steps. We used two sequence from A. chukar, A. rufa as IntroductIonThe barbary partridge Alectoris barbara is widespread in North Africa from Morocco to Egypt and is also present in Sardinia (Cramp & Simmons 1980, Madge & Gowan 2001). Over much of range it has declined due to hunting pressure and habitat degradation (Madge & Gowan 2001, BirdLife International 2004). Locally numerous in Moroc-co, Algeria and Tunisia, current status in Lybia is unknown and it is perhaps extinct in Egypt (Madge & Gowan 2001).The eastern subspecies A. barbara barbata, who lives in the Cyrenaica, was described as a species by Reichenow (1896) and later, as Caccabis callolaema, also by Salva-dori & Festa (1916). It was also considered as separate species by Ghigi (1923); however, in the recent literature, it is usually listed under subspecies of Alectoris barbara (Cramp & Simmons 1980, Madge & Gowan 2001).  The purpose of this paper is to provide some starting points for a correct systematic position of Alectoris barba-ta.MaterIal and MethodsIn addition to the obvious visual comparison of the mor-phological characteristics of the two taxa we have also measured some morphometric parameters (bill, wing, third On the systematic status of the Cyrenaic Partridge(Alectoris barbata Reichenow, 1896)Silvio Spanò1, irene pellegrino2, enrico Borgo31Università degli Studi di Genova, DIPTERIS - Corso Europa 26, 16132 Genova (spano.silvio@gmail.com)2Università del Piemonte Orientale, DISIT (Dipartimento di Scienze ed Innovazione Tecnologica) -Via Teresa Michel 11, 15121 Alessandria (irene.pellegrino@mfn.unipm.it)3Museo Civico di Storia Naturale “G. Doria” - Via Brigata Liguria 9, 16121 Genova (eborgo@comune.genova.it)abstract – Some considerations on the morphological features that differentiate Alectoris barbara barbata by A. b. barbara are exposed and are also reported the results of a genetic investigation performed on historical specimens. Results showed a considerable genetic dis-tance (0.60), certainly enough to consider it an ESU (Evolutionary Significant Unit), but most likely a separate species.Key-words: Alectoris barbara barbata, Cyrenaica, genetic.145Avocetta 37: 145-148 (2013)Spanò et al.streak, there is rarely only a vague hint of a second streak in some feathers (Fig. 2); different appearance of the neck, the eyebrows do not break the collar; even with a few spec-imens measured, the values of the considered morphomet-ric measures, in particular the tail, are significantly greater than in A. barbara (Tab. 1). Results of genetic test conducted on 4 specimens of A. barbara barbata led to the results shown in Fig. 3 (NJ tree with Tamura Nei distance on mtDNA d-loop, 241nu-cleotides) and in Tab. 2 (TN93 genetic distance). These results indicate that genetic distance between A. barbara from Sardinia and Tunisia and A. barbara from Cyrenaica was 0.060 (6%). outgroups and two sequence A. barbara from Sardinia and Tunisia as reference. results Alectoris barbara barbata presented several morphologi-cal characters rather different from the other subspecies of Barbary Partridge and in particular: throat and eyebrows dark bluish grey (Fig. 1); feathers of the throat with elon-gated flattened rachis, with “beard” effect; brown orange collar with greyish patches, not red-brown with white patches; feathers of the flanks with one single thick black 146Figure 1. Differences between Alectoris barbara barbata (left) and A. b. barbara (right) in the colour of collar and throat and in the shape of the feathers of the throat.Figure 2. Differences in the pattern of the feathers on the flanks between Alectoris barbara barbata (left) and A. b. barbara (right).On the systematic status of the Cyrenaic Partridge147dIscussIonGhigi in 1923 was probably the last to be interested in this taxon and he considered Alectoris barbata as a separate species. Having had the opportunity to hold in captivity several individuals of this partridge, he emphasized, in ad-dition to the morphological characters described above, al-so information on the behaviour that supported him in con-sidering A. barbata a separate species (Ghigi 1920, 1921, 1923). The high genetic distance 0.060 (6%), the morphologi-cal characteristics, and the statistically significant differ-ence in tail length found between the samples of the two taxa analysed indicates that Alectoris barbara barbata should be considered as a distinct evolutionary significant unit (ESU), as for A. graeca whitakeri, the Sicilian endem-ic subspecies of Alectoris graeca (Randi et al. 2003).  However we should take into account that the par-tridges are sedentary species and A. barbara barbata is en-demic to the Cyrenaica Peninsula in Northern Lybia and thus separated from the other North African populations of A. barbara. This isolation increased the morphologic and genetic differentiation that became so important that this taxon could be treated as a separate species, follow-ing the opinion expressed by Reichenow (1896, original description), Salvadori & Festa (1916) and Ghigi (1920, 1921, 1923). A closer examination should be carried on with a larg-table 1. Mean values (± SD) of the considered morphometric measures in individuals of Alectoris barbara barbara or A. barbara bar-bata and p values of the comparisons between the two subspecies (Mann-Whitney test).table 2. Tamura Nei distance (TN93) between analyzed taxa. Genetic distance between Alectoris barbara from Sardinia and Tunisia and A. barbara from Cyrenaica is 0.060 (6%). Standard error (in italic) is calculated on bootstrap methods with 1000 replicates.Figure 3. mtDNA d-loop (241 bp) Neighbor Joining tree with Tamura Nei distance (TN93). Four samples from Cyrenaica, Libya (4 indi-viduals sampled between 1915 to 1922, Regional Museum of Natural Sciences, Turin) were analyzed. Two sequence of A. barbara from Tunisia and Sardinia were used as the reference and two sequences of A. rufa and A. chukar were used as outgroups. Support for the in-ternodes was assessed by bootstrap percentages after 1000 resampling steps.BillIII PrimaryTarsusTail17.77 ± 2.48 (N = 10)104.94 ± 4.89 (N = 8)39.40 ± 3.15 (N = 10)88.70 ± 5.10 (N = 10)19.04 ± 1.06 (N = 12)111.17 ± 2.79 (N = 12)43.73 ± 2.69 (N = 11)106.33 ± 5.90 (N = 12)0.0120.0060.0030.00001A. b. barbara A. barbara barbata P (Mann-Whitney)A. rufaA. chukarA. barbara SardiniaA. barbara TunisiaA. barbara Cyrenaica-0.1070.1620.1620.1370.021-0.1180.1180.1380.0290.023-0.0000.0600.0290.0230.000-0.0600.0250.0250.0160.016-A. rufa A. chukar A. barbaraSardiniaA. barbaraTunisiaA. barbaraCyrenaica0.0196856397100A. Barbara 3A. Barbara 1A. Barbara Sardinia (Italy)A. Barbara TunisiaA. rufaCyrenaicaA. chukarA. Barbara 4A. Barbara 2Spanò et al.er number of specimens, preferably with fresh material. Moreover being an ESU-separate species, ecological and demographic investigation are needed to assess the status and monitor demographic trends of A. barbata.acknowledgements – We wish to thank Elena Gavetti and Marco Pavia (Museo Regionale di Scienze Naturali, Turin), Sylke Frah-nert (Museum fur Naturkunde, Berlin) and Giuliano Doria (Mu-seo Civico di Storia Naturale “G. Doria”, Genoa) for the courtesy and their help.reFerencesBirdLife International, 2004. Birds in Europe: Population Esti-mates, Trends and Conservation Status. BirdLife Internation-al, Cambridge.Cramp S. & Simmons K.E.L., 1980. Handbook of the Birds of Europe, the Middle East and North Africa, II. Oxford Univer-sity Press. Oxford.Felsenstein J., 1985. Confidence limits on phylogenies: an ap-proach using the bootstrap. Evolution 39: 783-791.Ghigi A., 1920. Vertebrati di Cirenaica raccolti dal prof. Ales-sandro Ghigi nella escursione organizzata dal Touring Club Italiano 15-24 aprile 1920. Memorie R. Accad. Sci. Ist. Bo-logna 6: 207-208.Ghigi A., 1921. La perdrix de la Cyrénaique (Alectoris callolaema Salvadori et Festa). Oiseau-Revue Hist. Nat. Appl. 2: 22-26.Ghigi A., 1923. Sulla pernice di Cirenaica Alectoris barbata (Rei-chenow). Riv. ital. Orn. 6: 3-7.Madge S. & McGowan P., 2002. Pheasants, Partridges e Grouse. Including Buttonquails, Sandgrouse and Allies. Christopher Helm, London. Randi E., Tabarroni C., Romindi S., Lucchini V. & Sfougaris A., 2003. Phylogeography of the rock partridge (Alectoris grae-ca). Mol. Ecol. 12: 2201-2214.Reichenow A., 1896. Uber ein merkwurdiges Steinhuhn. Orn. Mber. 4: 76.Saitou N. & Nei M., 1987. The neighbour-joining method: a new method for reconstructing phylogenetic trees. Mol. Biol. Evol. 4: 406-425.Salvadori T. & Festa E., 1916. Alcuni uccelli della Cirenaica, col-la descrizione di una nuova specie del genere Caccabis. Boll. Musei Zool. Anat. comp. R. Univ. Torino 31: 1-2.Tamura K. & Nei M., 1993. Estimation of the number of nucleo-tide substitution in the control region of mitochondrial DNA in humans and chimpanzees. Mol. Biol. Evol. 10: 512-526.Tamura K., Peterson D., Peterson N., Stecher G., Nei M. & Kumar S., 2011. MEGA5: Molecular Evolutionary Genetics Anal-ysis using maximum likelihood, evolutionary distance, and maximum parsimony methods. Mol. Biol. Evol. 28: 2731-2739.148

On the systematic status of the Cyrenaic Partridge (Alectoris barbata Reichenow, 1896)

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