ACTIVE AND PASSIVE HEALTH SURVEILLANCE IN A PYRENEAN CHAMOIS POPULATION

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1 Chamois International Congress Majella National Park Lama dei Peligni-Italy 17th-2th June, 214 ACTIVE AND PASSIVE HEALTH SURVEILLANCE IN A PYRENEAN CHAMOIS POPULATION Facultad de Veterinaria Universidad de Zaragoza C/ Miguel Servet, Zaragoza MaríaCruz Arnal 1,3, David Martínez 1, Miguel Revilla 1, Mª Jesús de Miguel 2, Pilar Mª Muñoz 2 & Daniel Fernández de Luco 1 1 Departamento de Patología Animal, Facultad de Veterinaria, Universidad de Zaragoza, Zaragoza, Spain 2 Centro de Investigación y Tecnología Agroalimentaria (CITA). Gobierno de Aragón, Zaragoza, Spain 3 Corresponding autor ( Pyrenean chamois health surveillance has been made in the Hunting Reserves (HRs) of the Aragonese Pyrenees (Fig.1) in a 14 years period, chamois ill or found dead have been analysed (Fig.2), in addition to 118 hunted animals and 2,61 samples of hunted chamois. The aim of the study was to establish the status of epidemic processes with serious impact on this species, including pestivirus disease, infectious keratoconjunctivitis, zoonoses such as brucellosis and tuberculosis; as well as pathological processes in ill or dead chamois found on the mountain. Los Valles HR Viñamala HR France Ordesa and Monte Perdido NP Los Circos HR Benasque HR Fig.1: Study area in the central and western Pyrenees (Aragon) divided in mountain massifs and showing the 4 Hunting Reserves (HR) and the Ordesa and Monte Perdido National Park (NP). Fig.2: Pyrenean chamois found dead or ill analysed per year. Mainly animals were analysed between due to the IKC outbreak and between due to the Pestivirus outbreak. Fig.3: Chamois affected by Pestivirosis The study of chamois Pestivirus Disease (Fig.3) was conducted in 2,426 chamois from Aragon (385 ill-dead and 2,41 hunted). In the Aragonese Pyrenees, neither infected chamois nor clinical signs were detected between 21 to July 211 (n=1,656). After the first detection in Aragon in 211 the virus was confirmed by Ag-ELISA in 97 out 77 studied animals (564 hunted and 26 ill/dead animals) and confirmed by RT-PCR in 28 animals. Serological studies against pestivirus (n=2,373) showed a higher proportion of seropositive animals in the affected HR in the last years (Fig.4). An outbreak of Infectious Keratoconjunctivitis (IKC) in chamois population in Aragon and Navarre was detected in the spring of 26 until late 28 (Fig.5). One hundred and nine affected animals were collected, mostly during 27 in Viñamala s HR and in 28 in Los Valles and Benasque s HRs. The process was seasonal, beginning in spring-summer and ending in winter, except in Benasque s HR where it went on until the following winter. M. conjunctivae was the main causal agent identified by rt-pcr in IKC affected chamois. It was identified in 89.42% of animals tested (n=14). A 4.21% (8/19) of hunted chamois, sampled during 26 to 29, without eye damage, were carriers of M. conjunctivae. Fig.4: Prevalence of pestivirus antibodies in Pyrenean chamois per year and HR. Fig.5: Adult, kid and yearling with keratoconjunctivitis. Fig.6: Chamois affected by fibrinous bronchopneumonia Fig.8: Purulent meningoencephalitis. Traumatic origin Fig.11: Lung with catharral bronchopneumonia A.5% (13/2,73) of Pyrenean chamois was seropositive against Brucellosis, being negative to bacterial culture. The necropsy of 49 ill-dead chamois revealed a 28.6% (117) with IKC, a 19.3% (79) with Pestivirus Disease, a 6.3% (26) with non-parasitic bronchopneumonia (Fig.6), 7% (29) with multiple traumatisms (Fig.8), 1.5% (6) with digestive processes, 1.7% (7) with other diseases (Contagious Ecthyma (Fig.7), cataracts and abscesses) and 35.4% (145) with inconclusive processes, being most of them incomplete animals. Other sporadic lesions observed were Dermatophilosis, pheochromocytoma, Hydatidosis and Pseudotuberculosis. Hunted animals (n=118) showed verminous pneumonia by small strongyles; coccidia, nematodes (Ostertagia spp. and Haemonchus spp. Fig.9) in the abomasum and Visceral Cysticercosis (Fig.1). The presence of areas with chronic catarrhal pneumonia in apical lobes (Fig.11) was frequent. No suspicious lesions compatible with either Tuberculosis (n =31) or Mange (n =299) were found in the Pyrenean chamois studied. Fig.7: Kid with Contagious Ecthyma Fig.9: Haemonchus spp. Abomasum *The authors acknowledge the dedicated assistance of the Rangers of Aragon. This work is financed by Aragon Government. Fig.1: Cysticercus tenuicollis

2 Osservazione della longevità degli esemplari di Camoscio Appenninico (Rupicapra pyrenaica ornata) rilasciati per la reintroduzione nel Parco Nazionale del Gran Sasso e Monti della Laga, e sopravvivenza al primo anno dei Kids nel periodo Carlo Artese, Gino Damiani - INTRODUZIONE La reintroduzione del Camoscio Appenninico nel Parco Nazionale del Gran Sasso e Monti della Laga è stata effettuata attraverso la liberazione di 26 esemplari dal 1992 al 1994 (Lovari, Artese, Damiani e Mari Global Reintroduction prospectives; IUCN 21) e successivamente durante il progetto Life "Conservazione di Rupicapra pyrenaica ornata" di altri 9 esemplari dal 1999 al 21 per un totale di 35 Camosci. Tutti gli esemplari erano dotati di marche auricolari e 17 anche di radiocollare VHF. (Lovari Mari relazione finale 24) Dal 1995 inizia la fase di formazione di branchi stabili e gli esemplari sono stati monitorati durante tutto l'anno. Vengono riportati i dato relativi a: 1. longevità e riproduzione di 23 dei 35 esemplari rilasciati ( 65,71%), presenti dal 1995 ad oggi, su una popolazione che nel 1995 contava circa 25 esemplari e nel 212 conta circa 45 esemplari; non sono stati considerati i dati degli esemplari morti prima del 1995 probabilmente correlati alla fase di reintroduzione dispersione e colonizzazione dei nuovi territori. 2. numero di Kids e Yearlings rilevati annualmente durante il monitoraggio primaverile estivo e, quando effettuato, il censimento autunnale; The reintroduction of the Apennine Chamois in the Gran Sasso-Laga National Park was realized through the release of a group of 26 chamois from 1992 to 1994 (Lovari, Artese, Damiani e Mari Global Reintroduction prospectives; IUCN 21) and afterwards during the Life Project "Conservation of Rupicapra pyrenaica ornata" of another group of 9 chamois from 1999 to 21 amounting to 35 chamois. All the individuals were equipped with earmarks and 17 of them also with WHF radio-collars. In 1995 the period of the formation of stable herds began and the individuals have been monitored throughout the whole year. We report here the data related to the longevity and the reproduction of 23 of the 35 released individuals (65,71%) that have been present from 1995 until today out of a population that counted about 25 individuals in 1995, about 38 individuals in 28 and about 45 individuals in 212. We documented the survival of 9 females, one of them survived for 2 years, one for 19 years and one for 18 years. The average lifespan calculated on all the earmarked females that are part of the present study is 13.7 years excluding one 15- year-old individual that is still alive. We documented as well the survival of 13 males, two of them survived 15 years, two 14 years, one 13 years, with an average lifespan of 9.4 years. The assumption of a highly significant relation between longevity and age of release was verified by Spearman's rank correlation coefficient r=,732 (p<.5) (p<.1). The longevity of the 1 females is significantly higher than the one of the 13 males (U=25,5; P<.5,Mann-Whitney U-Test). An average death rate of 1,6% per year has been calculated using a contingency table between the expected population (population without any phenomenon of emigration or immigration) knowing the annual "recruitment" (kids observed in July) and the population at the end of the year (estimated on the basis of the comparison between data obtained by monitoring in spring and the results of the block counts performed in autumn in the period 2/28). In the first year the survival rate (number of yearlings recounted the year after they were born) calculated during that period was %. We documented the reproduction of three individuals at the age of 16, 15 and 12 years. We notice how the longevity of the Apennine chamois population in the GSLNP, under conditions of maximum potentiality for the species, is a decisive factor of the reproductive success. We reaffirm the importance of long-term monitoring as an indispensable instrument for the understanding of the factors that contribute to the success of reintroduction. Area di studio Massiccio montuoso del Gran Sasso d Italia RISULTATI E' stata documentata la sopravvivenza di 9 esemplari di sesso femminile (una per 2 anni, una per 19 anni, una per 18 anni); tale longevità non è riscontrata in letteratura per il Camoscio Appenninico ma conosciuta per il Camoscio Pirenaico. La vita media di tutte le femmine marcate oggetto del presente lavoro è di 13,7 anni escludendo un esemplare di 15 anni ancora vivo. E' stata documentata la sopravvivenza di 13 esemplari di sesso maschile di cui due per 15 anni, due per 14 anni, uno per 13 anni con una vita media di 9,4 anni. L'ipotesi di relazione altamente significativa tra longevità ed età del rilascio è stata verificata attraverso il coefficiente di Spearman per ranghi con valore di r=,732 (p<.5) (p<.1). Si è successivamente ipotizzato che tale longevità potesse essere alla base del marcato incremento percentuale della popolazione pari al 23% annuo osservato dagli autori e pubblicato durante il secondo progetto Comunitario Life natura (Mari e Lovari 26). Attraverso una comparazione tra la popolazione attesa (popolazione senza fenomeni di emigrazione/immigrazione) conoscendo il "recruitment" annuale (Kids osservati nel mese di luglio) e la popolazione a fine anno, presunta dagli autori in base al raffronto dei dati "monitoraggio primaverile e censimento (blokcounts) autunnale nel periodo 2/28, si è calcolato un tasso di mortalità pari al 1,6 % annuo. Il termine recruitment indica non la vera e propria natalità (numero di capretti nati) ma quelli che, superando i primi due mesi di vita, vanno ad incrementare il branco. I motivi di questa scelta sono riconducibili alle grandi difficoltà di contattare le femmine nel periodo riproduttivo che, scegliendo per partorire le cenge più verticali e inaccessibili, di fatto rendono impossibile verificare nelle prime settimane l effettiva natalità. La percentuale di capretti morti per cause naturali durante il parto o nei giorni immediatamente successivi o predati nelle prime settimane viene dunque calcolata successivamente in maniera statistica facendo la differenza tra il recruitment e le femmine adulte presenti. Il dato dei capretti reclutati è sicuramente il più completo e preciso. Si conferma che in specie come il Camoscio a basso dimorfismo sessuale (con differenza di peso tra i sessi intorno al 1% del peso, il raggiungimento del peso ottimale al terzo anno di età e la struttura sociale matriarcale e poligama) la sopravvivenza dei maschi sia inferiore rispetto a quella delle femmine. La longevità dei 1 esemplari di sesso femminile è significativamente maggiore di quella dei 13 esemplari di sesso maschile (U=27,5; P<.5,Mann-Whitney U-Test). Il tasso medio di sopravvivenza al primo anno dei Kids è, nel periodo , pari all 84,92%. anno recruitmentyerling % , , , , , , rosso = F - nero = M Confronto attesa e presunta dagli autori popolazione attesa stima non statistica fit dei dati sulla popolazione attesa con riferimento al modello Malthusiano coefficiente di correlazione y = 6,8141e,2526x R 2 =, longevità FemmineMaschi ,7475 9,21 ultima natalità osservata ANNO DI ultima NOME NASCITA natalità BELLA ANNALINA COCCINELLA FINALMENTE GIOVANNA ORNELLA MOSTRO ROMY VERA NETZ E' stata documentata la riproduzione degli esemplari: Bella e Giovanna a 16 anni, Annalina a 15 anni, Netz a 12 anni. METODO Si fa riferimento al metodo cattura marcatura ricattura (Cormack 1964) considerando come ricattura ogni localizzazione del singolo esemplare marcato. L'età è stata ricavata dalla conoscenza della data di nascita per animali provenienti dalla cattività o dall analisi dell'accrescimento corneo al momento della cattura. Il set temporale di dati relativo alle localizzazioni degli esemplari marcati è di circa 13 anni. I dati raccolti con metodo naturalistico sono stati valutati attraverso test statistici non parametrici. La popolazione di camoscio del Gran Sasso è soggetta a predazione da parte del Lupo e dell'aquila reale. La dispersione per la specie, in particolare per le femmine adulte, è molto rara e la specie è prevalentemente sedentaria (Lovari e Locati 1991); non vi sono ancora fenomeni di immigrazione o emigrazione da parte di altre colonie e la popolazione non è soggetta prelievo venatorio. Essendo gli esemplari rilasciati aggregati o circoscritti a sole tre località (carta area di studio) la contattabilità è stata costante e nei primi anni uguale al 1% degli animali presenti; la media delle localizzazioni per singolo esemplare marcato nei tredici anni di monitoraggio del presente lavoro è stata di 24,2; non essendo però stato possibile per ovvi motivi per molti esemplari conoscere la data esatta di morte, il gruppo di lavoro addetto al monitoraggio, ha ritenuto di far coincidere la data di morte con l'inverno successivo l'ultima localizzazione. anno di anno di momento del ultima MORTE MORTE N NOME MARCA DX MARCA SX nascita SESSO rilascio rilasacio localizzazione ACCERTATA PRESUNTA LONGEVITA 5 BELLA gialla 2 gialla F feb ANNALINA gialla 5 gialla F ott COCCINELLA rosso 2 rosso F lug FINALMENTE gialla F gen GIOVANNA rossa 7 sette 1988 F apr ORNELLA bianca 4 bianca F set MOSTRO gialla F gen ROMY v erde 2 verde F ott VERA rosso 3 verde F apr NETZ giallo rosso 1998 F ott FERNANDO v erde B verde B 1987 M gen TEX rosso A rosso A 1987 M ago APPOLONIO barra due 1992 M nov GANDALF due punti due punti 1988 M nov RIPIDO triangolo triangolo 1992 M mar LORETO rosso 1998 M ott SALVO punto 1995 M nov SVELTINO croce croce 1999 M mag AMERICO rettangolo 2 M giu BELLINI stella stella 1999 M ott CARISSIMO zero zero 2 M ago FEDERICO doppia 2 M ago FIOCCO freccia freccia 1994 M mar eta al DATA DI DATA DI DISCUSSIONE 1) La longevità della popolazione di Camoscio Appeninico nel PNGSL, in condizioni di massima potenzialità per la specie anche in presenza di predatori, è un fattore determinante del trend positivo? 2) La diminuzione della capacità riproduttiva con l'aumentare dell'età, fenomeno di senenscenza evidente per gli esseri umani, non è univoca e empiricamente provata per il genere Rupicapra. Le osservazioni riportate di alta longevità delle femmine e buon numero medio di riproduzioni per femmine rilasciate, alta sopravvivenza dei kids e basso tasso medio di mortalità sono alla base del successo della reintroduzione sul Gran Sasso? 3) I monitoraggi a lungo termine (quasi mai programmati e attuati dagli Enti gestori delle reintroduzioni) sono uno degli strumenti indispensabili per la comprensione dei fattori di riuscita di una reintroduzione? Si ringraziano i colleghi e i collaboratori del Servizio Scientifico dell Ente e il Direttore del Parco Dott. Marcello Maranella

3 Group dynamics and local population density of Apennine Chamois at the Abruzzo, Lazio and Molise National Park: trend and spatial variation Asprea A., Pagliaroli D. & Latini R. Servizio Scientifico del Parco Nazionale d Abruzzo, Lazio e Molise INTRODUCTION AND AIMS The social organization of a species may be influenced by various factors. As a general rule, in ruminants number, size and composition of groups should vary according to population density. Recent studies on the Apennine Chamois (Rupicapra pyrenaica ornata) at the Abruzzo, Lazio and Molise National Park (hereafter abbreviated as PNALM) showed that this population may be subjected to densitydependent processes. Over the last 8 years, population size has generally decreased, so we expect corresponding changes in social structure, particularly in group number and mean group size. To test this hypothesis we analyze the detailed summer data on group size and composition collected in and those collected in when the population trend was opposite. MATERIAL AND METHODS Standardized repeated visual scans performed along the same routes during which number, size and location of chamois groups were recorded (mean N ± sd: 11 ± 5 repeats; 18 ± 42 groups sighted per year). Group was defined as one or more individuals close to each other and located at least 5 meters from other individuals. Group size classes were defined as follows: 1, 2-5, 6-1, 11-2, 21-4, >4 individuals. Sex and age class of each individual were also noted. Local population density calculated as the number of individuals seen in each scan session in the 1% MCP area based on the locations of all the chamois groups sighted during the repeated counts. Raw data were log-transformed. 1) Ln (local density) 4, 3,8 3,6 3,4 3,2 3, 2,8 2,6 STUDY AREA Val di Rose (ca. 3.5 km 2 ) is located in the core of the PNALM and hosts one of the most representative chamois herds. The area is attended also by red deer (Cervus elaphus) and wild boar (Sus scrofa). Domestic livestock is not present. Local population density dropped from 83.4 to 21.7 heads/km 2. Mean F(7;82) = 4.1, P <.1; Spearman, r = -.45, P <.1 Mean group size decreased. Mean±,95 Conf. Interval RESULTS Ln (N groups) 2,8 2,6 2,4 2,2 2, 1,8 1,6 1,4 Mean Group number increased. Mean±,95 Conf. Interval Kruskall-Wallis, df = 7, H = 21.8, P <.1; Spearman, r =.27, P <.5 2) Ln (N groups) 4) Positive correlation between local population density and both mean group size (Spearman, r =.43, P <.1) and group number (Spearman, r =.27, P <.5). Population density accounted for 18.4% and 14.1%, respectively, of their variation (GLM, P <.1). 3,6 3,2 2,8 2,4 2, 1,6 1,2,8,4 2, 2,4 2,8 3,2 3,6 4, 4,4 4,8 Ln (local population density) Ln (N groups) = *x;.95 Conf.Int.; Spearman, r =.27, P <.5 The number of groups >2 was positively correlated with population density (Spearman, r =.29, P <.5) and negatively correlated with female group number (Spearman, r = -.42, P <.1). Ln (mean group size) 3,6 3,2 2,8 2,4 2, 1,6 1,2,8 2, 2,4 2,8 3,2 3,6 4, 4,4 4,8 Ln (local population density) Ln (mean group size) = *x;.95 Conf.Int.; Spearman, r =.43, P <.1 Female group number was negatively correlated with mean group size (Spearman r = -.4, P <.1) and explained the 22.8% of its variation (GLM, P <.1). Ln (mean group size) 3) 2,6 Mean Mean±,95 Conf. Interval 2,4 2,2 2, 1,8 1,6 1,4 1,2 1, F(7;82) = 1.8; P <.1; Spearman, r = -.66, P <.1 Strong concordance in group frequency distribution across years (Kendall Concordance Coefficient, W =.91). Singleton class was the most frequent in , whereas 2-5 class was most frequent in The number of groups > 2 individuals decreased (Spearman, r = -.81, N = 8, P <.1) and the number of groups with 1-5 individuals increased (Spearman, r =.78, N = 8, P <.5). 4,8 3,6 4,4 3,2 Ln (local population density) Ln (groups with F Ad) 4, 3,6 3,2 2,8 2,4 2, -,5,,5,1,15,2,25,3,35,4,45 Ln (% groups with N >2) 3, 2,5 2, 1,5 1,,5, -,5 -,5,,5,1,15,2,25,3,35,4,45 Ln (% groups with >2 individuals) Ln (mean group size) 2,8 2,4 2, 1,6 1,2,8 -,5,,5 1, 1,5 2, 2,5 3, Ln (% groups with F Ad) DISCUSSION Overall, our results: outline the complex relationship between social structure and population density and are consistent with what generally reported in literature; support the hypothesis that the population density decrease was related to an increase in small groups and a decrease in medium-large groups, in other words to a more scattered distribution of chamois in the area. However, Val di Rose does not seem to be representative of the whole PNALM, since the analysis of data collected in in five areas, including Val di Rose, suggest that a certain spatial variability exists. These samples are too small to perform statistically reliable tests on trends, nevertheless each year the frequency distribution of group size showed quite a low concordance among herds (Kendall Concordance,.54 U.7). Thus, density-dependent processes might differ from place to place and/or each herd might respond differently in relation to local conditions. Acknolowdgements We are grateful to D. Russo for useful comments on the work. LIFE9 NAT/IT/183 Development of coordinated protection measures for Apennine Chamois (Rupicapra pyrenaica ornata) COORNATA Chamois International Congress Lama dei Peligni (AQ) - June, 17-19, 214

4 Chamois International Congress - Maiella National Park, Lama dei Peligni, Abruzzo, Central Italy - 17 th -2 th June 214 Mass mortality by lightning in Apennine chamois (Rupicapra pyrenaica ornata): a case report from the Abruzzo Lazio e Molise National Park, Italy Badagliacca Pietro 1*, Gentile Leonardo 2, Marruchella Giuseppe 1, Latini Roberta 2, Di Pirro Vincenza 2, Carosi Emiliano 2, Ruberto Addolorato 1, Scioli Erminia 3, Di Provvido Andrea 1 1 Istituto Zooprofilattico Sperimentale dell Abruzzo e del Molise G. Caporale, Campo Boario, 641 Teramo, Italy 2 Ente Parco Nazionale d'abruzzo Lazio e Molise, Viale Santa Lucia, 6732 Pescasseroli, Italy 3 Azienda Sanitaria Locale 1 Abruzzo, Servizio Igiene Allevamenti e Produzioni Zootecniche, Via Umberto I, 6731 Castel Di Sangro, Italy * Corresponding author. Mailing address: Campo Boario, 641 Teramo, Italy; Phone ; Fax ; Introduction Lightning is a weather-related phenomenon capable of causing injury or death to people and animals, due to electrical effects, heat production and concussive force [1, 2]. Actually, the exact pathophysiology of lightning injury is not well understood because of the large number of variables that cannot be measured or controlled when an electrical current passes through tissue. However, death is usually caused by acute cardiac failure and/or by the arrest of the respiratory center in the brainstem. In addition, heating of tissues secondary to high-voltage current are known to cause characteristic linear skin burns (so-called lightning figure ) [3, 4]. The present report aims at describing a mass mortality event in Appennine chamois (Rupicapra pyrenaica ornata) due to lightning, thus focusing on that uncommon threat for wildlife. History, clinical signs, pathological and laboratory findings At present, the entire population of Apennine chamois consist of about 1,7 head, 4 of which reside within the Abruzzo Lazio e Molise National Park (ALMNP). On April 23 rd 214, after a hiker s alert, ten Apennine chamois were found dead in locality Monte Sterpi d Alto (Civitella Alfedena, L Aquila, Italy) by the personnel of the ALMNP (see Figures 1 and 2 for further details). Chamois carcasses three males and seven females, aged between one and nine years were inspected, individually identified (n. 1 to 1), and then referred to the Istituto Zooprofilattico Sperimentale Abruzzo e Molise G. Caporale (IZSAM) for detailed necropsy and laboratory diagnostic investigations. Field observations and necropsy pointed out scorched streaks on the skin surface (Figures 3-5), which were severe in two chamois (n. 2 and 9) and mild in other three head (n. 6, 8 and 1). Subcutaneous oedema and petechiae, pulmonary oedema, foci of parasitic bronchopneumonia, and congestion of the small intestine were additional consistent findings (Figures 6-9). Bacteriological, virological, parasitological and histopathological investigations yielded inconclusive diagnostic results. On the basis of what above, the diagnosis of lightening was made. Figure 1. Concerned area and geographical position of the animals*. All carcasses were found within the area delimited by the red-dotted line, which was about 548 m 2 large. Five chamois with skin scorched streaks (n. 2, 6, 8, 9 and 1, all marked in red) were very close each other and placed along an imaginary north-west/south-east line. The distance between chamois n. 2 and n. 8 was about 33 m. Two chamois (n. 4 and 5) laid near that line, but with no apparent skin lesion. The remaining three chamois (n. 1, 3 and 7) were found further down the slope, the distance between n. 1 and 8 being 43.7 m. Figure 3. Chamois n. 2. A streak is clearly seen on the skin surface, running along the left side of the neck, the upper chest and the abdomen. The so-called step potential can occur when animals are standing along the direction of potential gradient. In that case, ground current flows via the legs and can cross the heart and other vital organs. Figure 4. Chamois n. 2. Corneal burn and neck lesion by lightning. This picture remarks that the electrical current can enter through the cranial openings (orbits, mouth, nose), and can directly injury the brainstem. * Geographical coordinates were derived by projection ED_195_UTM_Zone_33N. The background map was obtained by ESRI DigitalGlobe service. Figure 2. Chamois carcasses found on the summit of Monte Sterpi, at an altitude of 196 m above the sea level. Figure 5. Chamois n. 9. Skin lesions are linear, hairless and dark grey in color. Figure 6. Corresponding to scorched streaks, subcutaneous oedema and hemorrhages were occasionally observed. Figure 7. The congestion of the small intestine was a consistent finding in all chamois under study. Figure 8. Foci of parasitic bronchopneumonia are a very common finding in Apennine chamois. Figure 9. Tapeworms were found within the intestinal lumen of few chamois. References 1. Cooper M.A., Andrews C.J. & Holle R.L. 27. Lightning Injuries in Wilderness Medicine, Auerbach, Mosby. 5 th ed. 2. Gomes C Lightning safety of animals. International Journal of Biometeorology, 56(6), DOI 1.17/s Mandelli G. & Finazzi M Apparato tegumentale. Cute, sottocute e annessi. In Guarda F., Mandelli G. & Biolatti B. (eds.), Trattato di Anatomia Patologica Veterinaria. 4 th Ed., Utet Medica, Torino. 4. Žele D., Bidovec A. & Vengušt G. 26. Atmospheric flash injuries in roer deer (Capreolus capreolus). Acta Veterinaria Hungarica, 54(1), Discussion and conclusion In veterinary medicine, lightning is mainly reported in domesticated animals housed outdoor, whereas it seems a rather uncommon and likely underestimated cause of death in free-living animals. Reasonably, wild animals are often exposed to lightening, but their carcasses are uncommonly found. Considering that carcasses with skin lesions were very close to each other and placed along the North-West/South-East direction, we hypothesize that a step potential/side flash combination killed those chamois. The remaining chamois, found outside that direction with no evidence of scorched streaks, were probably hit by a minor electrical current or side flash, which allowed them to move before dying. Monitoring the health status of endangered wild species is crucial for their preservation. Necropsy findings were decisive in order to solve the present diagnostic query. The same pathological findings remark that lightening should be always considered as differential diagnosis in the course of mass mortality event affecting wild animals. Finally, the presence of severe pulmonary and intestinal parasitic lesions confirms a worrying health situation for the source-population of Apennine chamois. Acknowledgement Our thanks go to Sandro Pelini, Paola Di Giuseppe and Gianfranco Romeo for their good technical support.

5 IZSTO Istituto Zooprofilattico Sperimentale del Piemonte, Liguria e Valle d Aosta SANITARY MONITORING OF THE ALPINE CHAMOIS (Rupicapra rupicapra) IN THE PROVINCE OF IMPERIA, ITALY (22-212) Maria Cristina Bona, Serena Durante, Maria Silvia Gennero, Maria Cesarina Abete, Simona Zoppi, Alessandro Dondo, Carla Grattarola, Maria Goria, Stefania Squadrone, Marco Ballardini, Giuseppe Audino, Giuseppe Ru, Walter Mignone * Istituto Zooprofilattico Sperimentale del Piemonte, Liguria e Valle d'aosta (IZSPLV), Wildlife Technician *Corresponding author: Introduction. For several years, the alpine chamois (Rupicapra rupicapra) population in the Italian Northwestern province of Imperia have been submitted to health monitoring. The monitoring was designed to investigate if coexistence between free-range livestock and wildlife in the area may have any effect. Alpine chamois in the Ligurian Alps represent the westernmost and southernmost population of this species in the Alps, with a particular interest because its habitat is influenced by the Mediterranean climate. During the hunting seasons, all hunted chamois are submitted to viscera examination, as well as to sampling of organs and tissues for chemical, bacteriological and virological analysis. Hunters are also requested to take a blood sample from each carcass, for serological investigation. Aim of this study is to provide the results of the monitoring activity carried out on the chamois between 22 and 212 in the Ligurian Alps. Material & Methods. Over the period 331 chamois were examined through the monitoring carried out in the Ligurian Alps (Figure 1). Both sampling data and the results of the analyses performed at IZSPLV were collected in an ad hoc database. The impact of diseases and chemical contaminants on the chamois population was evaluated in term of case prevalence checking for potential statistical association between disease and age and sex. Regional Park of the Ligurian Alps prev* parasitic pneumonia cysticercosis pseudotbc Fig 2. Prevalence of anatomopathological lesions Results.Out of the 313 animals submitted (Figures 2 and 3), 18 showed parasitic pneumonia (prevalence %CI ), 63 cysticercosis (2.1 95% IC ) and 2 pseudotuberculosis (6.4 95% IC ). The probability of the first two diseases was statistically higher in the oldest animals. Among undetected disease there were: Brucellosis, Paratuberculosis, Blue Tongue, Border disease, Yersinia spp., Francisella Tularensis and Toxoplasmosis.Four cases of keratoconjunctivitis by Mycoplasma conjunctivae (Figure 4), have been detected in 26 with a prevalence of 2.4% (IC 95%.7 6.). The presence of heavy metals (Pb, As, Cd, and Cr) showed very low values (Median As:<.1; Cd:.4; Cr:<.5; Pb:.5,Figure 5). Fig 4. Chamois with keratoconjunctivitis Density Cadmium (liver) Fig 5. Distribution of cadmium values in liver Fig 3. Liver with pseudotuberculosis Conclusion. The data allow the detection of infectious diseases circulating within the local chamois population and potentially having an impact on human and/or domestic animals health, or on the chamois population itself. Results obtained during this study do not highlight any specific problem with the coexistence between free-range livestock and wildlife in the area. The presence of the pseudotubercolosis could arise concern for the following two reasons: the widely spread of the pathology in the Alps and lack of measures of prophylaxis in the domestic animals. Chamois International Congress Majella National Park - 17 th -2 th June, 214

6 HEY, DO YOU WANT TO TASTE? USE OF BAITED BOX TRAP BY APENNINE CHAMOIS Bonanni M. 1, Scillitani L.1, Cobre P.1, Riganelli N.12, Artese C.12, Damiani G.12 1 Life + Natura Project Coornata LIFE9 NAT/IT/183 2 Parco Nazionale del Gran Sasso e Monti della Laga, Via del Convento 671 Assergi In the Gran Sasso e Monti della Laga National Park, for the first time after the reintroduction, captures of free ranging Apennine chamois (Rupicapra pyrenaica ornata) were performed, developing and testing new capture methods, in the frame of the LIFE project: Coornata: development of coordinated protection measures for Apennine Chamois (LIFE9 NAT/IT/183). Captures aimed at marking individuals within the protected area to better understand their spatial behaviour and response to capture stress and to translocate chamois to create two new colonies in the neighbouring protected areas involved in the project. Two different type of traps were employed. A larger one partially constructed of wood was built into a natural cave, it measures about 1.5 m x 3. m x 2. m. The smaller trap consist of a metal frame covered with metal wire mesh measuring 1.5 m x 2. m x 1.7 m. Both traps were provided with a drop gate at one end remotely controlled. We set both traps in Monte Coppe (42 26'19.85''N 13 45'3,45''E) in areas of observed high chamois use.the larger trap was built in October 21, baited with salt, concentrated vegetable attractants and fruits. Entrance in the trap and utilization of salt lick was monitored through photos and videos acquired by camera trapping. We observed a progressive increase in the number of visits to the box trap within the study period: in 21 we recorded 2 visits in 3 months, in visits, 31 in 212, 48 in 213 and 2 in 214 (from January to March). Chamois spent a minimum of 1 minute inside the trap to a maximum of 11 hours inside or in the immediate neighbourings of the trap. As for the number of visits, we observed a progressive increase in the mean time interval spent by chamois inside (or in the proximity of) the box trap throughout the study period: from 29 minutes in 211 to 134 min in 213. n occasion Seasonal time of entrance in the box time slot inverno primavera estate autunno We captured and ear tagged 3 chamois in june-july 211. Only two marked animals (1 male and 1 female) were camera trapped inside the box trap in the study period, both re-entered the trap exactly one year after being trapped. While the female entered again only in one occasion, the male entered 4 times, once 3 years after being captured. min Mean time interval spent inside the trap The trap was visited more frequently in autumn (5 occasions), spring (34) and winter (23), while we detected, in the whole study period, only 5 occasions in summer, as expected since the Monte Coppe area is mainly a wintering area, while in summer chamois groups move to higher elevation in neighbouring peacks. Chamois entered inside the trap more frequently during daylight, in spring and summer chamois never entered in the trap during night. Instead we recorded entrances within all time slots in autumn and in winter. We recorded the presence of all age and sex classes in the box-trap, even if yearlings and young males were the most represented category. We recorded the presence of kid only in one occasion. In most occasions (59%) only a single animal at a time visited the trap, in 31 (28%) occasions 2 animals entered together, and we recorded a maximum of 6 chamois inside the trap (1 occasion). The smaller box trap was built in November 212. During winter 212/213 it was damaged by snow and wind, and it was fixed in April 213. We used salt for bait.in late spring 213 we observed a high use by chamois. However it was not possible to compute the same analysis made for the larger box trap. Anyway in that period we had direct observations and capture data. The box trap was mainly visited by young individuals (yearlings and class I individuals- 2 to 3 years old) in late spring, while in autumn also older individuals entered to lick salt. In this box trap we caught a total of 5 chamois (2 males and 3 females) in 213 and 2 chamois (1 male and 1 female) in 214. Chamois International Congress Lama dei Peligni (CH), Parco Nazionale della Majella, 17-2 giugno 214 Aknowledgements: We thank for the collaboration in fieldwork activities: Ilaria Angelini, Paolo Montanaro and Corpo Forestale dello Stato (CTA) officers.

7 1 Research Station and Museum, Slovak National Forests TANAP; Slovak Republic; 2 Institute of Parasitology Slovak Academy of Sciences; Slovak Republic; 3 Tatra National Park, Poland; Tatra chamois (Rupicapra rupicapra tatrica Blahout, 1972) is a significant representative of the Tatra endemic fauna species that has been classified as critically endangered. According to the annual count in 213, the population of chamois in the territory of the Tatra Mountains represents 872 individuals in the Slovak Tatra National Park (TANAP) and 314 individuals in the Polish Tatra National Park (TPN). The numbers and health status of these animals is affected by plurality of different critical factors from climatic conditions through industrial pollution of the environment to individual anthropogenic impacts. In terms of health hazard, a significant role play parasitic infections that can lead to significant depletion of the entire population. The research on parasite fauna of Tatra chamois in Slovakia took place for several decades, but it was targeted only on the incidence of lung nematodes, which were considered the greatest threat. In contrast, the most recent research focused on gastrointestinal parasites was conducted in early 8 s of the last century (Mituch et al., 1984). Thus, the aim of our work was to study the occurrence of gastrointestinal parasites of Tatra chamois in current environmental and climatic conditions. MATERIAL AND METHODS Faecal samples from Tatra chamois (Rupicapra rupicapra tatrica) examined for propagation stages of gastrointestinal parasites. Sheather's and Faust's Flotation methods used. 164 samples from 24 localities of High Tatra Mountains (HTM) Additionally: 15 samples from West Tatra Mountains (WTM) 33 samples from Polish Tatra Mountains (TPN) RESULTS Slovak Tatra Mountains (TANAP) High Tatra Mountains (HTM) Examined: 164 samples Eimeria spp % Parasite-positive: 96 Moniezia spp % Prevalence: 58.5 % Trichostrongylidae 9.1 % Nematodirus spp.1.2 % West Tatra Mountains (WTM) Capillaria spp. 2.4 % Examined: 15 samples Parasite-positive: 2 Polish Tatra Mountains (TPN) Eimeria spp. 1 sample Trichostrongylidae: 1 sample Examined: 33 samples Eimeria spp % Parasite-positive: 14 Moniezia spp % Prevalence: 42.4 % Trichostrongylidae 9.1 % The initial research on gastrointestinal parasites of the Tatra chamois introduced one disputable finding - a relatively high prevalence of the genus Moniezia. In the Slovak part of the High Tatra, nearly 3 % of samples were positive for the tapeworm eggs that is significantly more in comparison with other European studies (see Stancampiano et al., 21; Hoby et al., 26; Morrondo et al., 21; Marreros et al., 211). These differences may be related to the presence of suitable intermediate hosts (Oribatida mites) in the environment, which is evidently closely linked to the climate and microclimate conditions - oribatid mite community composition would show a strong response to changes in habitat and land-use. Research initiated in the High Tatra after the huge windstorm in 24 revealed an increase in density of soil mites (and overpopulation of Oribatida mites), but also the decline in species diversity (Kalúz et Ferenčík, 28). All the above mentioned points to the need of continuing intensive research on parasite composition and distribution in Tatra chamois, in particular broader temporal, ecological and zoological contexts. This study was supported by bilateral Slovak-Polish project APVV SK-PL

8 The use of block-counts, mark-resight and distance sampling to estimate population size of Alpine chamois Luca Corlatti 1,2 *, Lorenzo Fattorini 3 & Luca Nelli 4 1 Institute of Wildlife Biology and Game Management, Vienna (A) 2 Wildlife Research Centre, Gran Paradiso National Park (IT) 3 Department of Economics and Statistics, Siena (IT) 4 Department of Earth and Environmental Sciences, Pavia (IT) * Correspondence: Introduction Population size estimates represent indispensable tools for many research programs and for conservation or management issues. Mountain-dwelling ungulates living in open areas are often surveyed through ground counts that, however, neglect detectability and normally underestimate population size (Loison et al. 26). While the use of sample counts is desirable, few studies have concurrently compared different probabilistic approaches to estimate population size in this taxon. Aim: taking advantage of a sample of marked individuals, we aimed to compare the size estimates of a male population of Alpine chamois Rupicapra rupicapra obtained with mark-resight and line transect sampling methods, using block counts to get the minimum number of males alive in the study area. Methods Between August-September 213, within the Gran Paradiso National Park (Italy) we performed independent surveys using: - block counts along purposely selected paths and vantage points; - mark-resight over 5 consecutive resightings from vantage points and paths; we analysed data using the Bowden s estimator (Fattorini et al. 27); - line transect sampling along 12 transects repeated 8 times; we analysed data using the Conventional Distance Sampling engine in Distance 6. (Thomas et al. 21) (figure on the right). Results & Discussion The figure on the left shows that: - block counts yielded a minimum number of males alive in the population of N=72 individuals; - the N=72 value was greater than the upper bound of the 95% confidence interval achieved using line transect sampling (N=54, SE=14%, 95% CI: 4-71); - mark-resight yielded a more realistic result of N=93 individuals (SE=18%, 95% CI: ). Line transect sampling performed poorly in the Alpine environment, leading to underestimates of population size, likely due to violations of some assumptions imposed by the rugged nature of the terrain. The mark-resight yielded lower precision, but likely provided robustness and accurate estimates since marks were evenly distributed among animals (Fattorini et al. 27). References Fattorini, L., Marcheselli, M., Monaco, A., Pisani, C., 27. A critical look at some widely used estimators in mark-resighting experiments. J. Anim. Ecol. 76, Loison, A., Appolinaire, J., Jullien, J.-M., Dubray, D., 26. How reliable are total counts to detect trends in population size of chamois Rupicapra rupicapra and R. pyrenaica? Wildl. Biol. 12, Thomas, L., Buckland, S.T., Rexstad, E.A., Laake, J.L., Strindberg, S., Hedley, S.L., Bishop, J.R.B., Marques, T.A., Burnham, K.P., 21. Distance software: design and analysis of distance sampling surveys for estimating population size. J. Appl. Ecol. 47, 5-14.

9 Physiological response to etho-ecological stressors in male Alpine chamois: timescale matters! Luca Corlatti 1,2 *, Rupert Palme 3 & Sandro Lovari 2 1 Institute of Wildlife Biology and Game Management, Vienna (A) 2 Department of Life Sciences, Siena (IT) 3 Department of Biomedical Sciences/Biochemistry, Vienna (A) * Correspondence: Introduction Faecal cortisol metabolite (FCM) levels have been widely used as physiological indicators of stress in a number of species (Möstl et al. 22). Integrating different sources of stress through an adaptive feedback mechanism, glucocorticoids may have important consequences on individual fitness, impacting on survival and reproductive success (Sapolsky 1992). To date, however, few studies have explored the role of proximate mechanisms responsible for the potential trade-offs between physiological stress and life history traits integrating social and environmental stressors. Aim: to investigate the variation in the effect of potential etho-ecological stressors (age, social status territorial vs. non-territorial males, minimum ambient temperature, precipitation, snow depth) on FCM excretion in male Alpine chamois over different timescales (year, cold months, spring, warm months, rutting season). Methods Between January 211 and December 212, in the Gran Paradiso National Park, we collected 393 scats on a monthly basis, for as many males as possible within a given month. Samples were analysed in duplicate SPRING using an 11-oxoaetiocholanolone enzyme immunoassay (Möstl et al. 22). RUT We used a model selection approach to analyse the effect of potential etho-ecological stressors (age, social status, minimum temperature, snow depth, precipitation) on FCM variation. Results & Discussion RUT - Over the year, FCM levels showed a negative relationship with minimum temperature, but altogether climatic stressors had negligible effects on glucocorticoid secretion, possibly owing to good adaptations of chamois to severe weather conditions; - age was negatively related to FCM during the rut (figure on the right), possibly due to greater experience of older males in agonistic contests; - social status was an important determinant of FCM excretion (figure on the left): while both the stress of subordination and the stress of domination hypotheses received some support in spring and during the mating season, respectively, previous data suggest that only the latter may have detrimental fitness consequences on male chamois (Corlatti et al. 212). References Corlatti L, Bethaz S, von Hardenberg A, Bassano B, Palme R, Lovari S (212) Hormones, parasites and alternative mating tactics in Alpine chamois: identifying the mechanisms of life history trade-offs. Anim Behav 84: Möstl E, Maggs JL, Schrötter G, Besenfelder U, Palme R (22) Measurement of cortisol metabolites in faeces of ruminants. Vet Res Commun 26: Sapolsky RM (1992) Neuroendocrinology of the stress response. In Becker JB, Breedlove SM, Crews D (eds) Behavioral endocrinology. MIT Press, Cambridge, Massachusetts, pp

10 Prevalence of gastrointestinal parasites in free range sheep, goats and cattle in the areas of presence of the Apennine Chamois (Rupicapra pyrenaica ornata) in five protected area in Central Apennine in Italy G. Cotturone 1, L. Gentile 2, U. Di Nicola 3, F. Morandi 4, S. Angelucci 5, M. Innocenti 3,5, V. Di Pirro 2, A. Argenio 2, E. Carosi 2, R. Latini 2, S. Gavaudan 6, F. Barchiesi 6, P. Morini 2, F. Striglioni 3, A. Rossetti 4, G. Damiani 3 M. Scacchia 7 1 Sirente-Velino Regional Park, 2 Abruzzo, Lazio and Molise National Park, 3 Gran Sasso and Mountain of Laga National Park, 4 Sibillini Mountain National Park, 5 Majella National Park 6 IZS Umbria and Marche, 7 IZS Abruzzo and Molise G. Caporale. INTRODUCTION Parasite infestation is one of the most common problem affecting cattle sheep and goats of all ages and breeds (Rafiullah et al., 211; Awraris et al., 212; Tshering et al, 213). Internal parasites interfere with nutrition, growth and the production (Pilarczyk et al., 29; Awraris et al., 212; Khan et al., 213, Tshering et al, 213). MATERIALS AND METHODS Study area Our study area was five protected areas in Central Apennine. 1. Abruzzo, Lazio and Molise National Park (PNALM): n hectares; 2. Majella National Park (PNM): n ; 3. Gran Sasso and Mountain of Laga National Park (PNGSL): n hectares; 4. Sirente-Velino Regional Park (PRSV): n hectares; 5. Sibillini Mountain National Park (PNMS): n hectares. Animal sampling A total of n. 916 fresh faecal samples were collected from of cattle, sheep and goat, including improve young, improved adult, native young and native adult from july 211 to april 214. These samples were collected just after defecation, using simple circular random sampling method. Faecal samples were placed into vial placed into cool box and transported for the laboratory examination and processed by standard floatation method to identify endoparasite species. RESULTS In cattle (319 faecals samples) microscopic examination revealed that about 165 samples (51.7%) were infected with gastrointestinal parasites. Among parasitic infectioned samples, helminth (51.7 %) and protozoa (28.2%) were examined. Strongyle (gastrointestinal) was the most common parasite in faecal samples of all cattle sampled (43.7%, n. 139). In sheep and goat (597 faecals samples) microscopic examination revealed that about 556 samples (93.1%) were infected with gastrointestinal parasites. Among parasitic infectioned samples, helminth (85.3%) and protozoa (74.1%) were examined. Endoparasites prevalence in cattle Endoparasites prevalence in sheep and goat 1,% 9,% 9,% 8,% 8,% 7,% 7,% 6,% 6,% 5,% 5,% 4,% 4,% 3,% 3,% 2,% 2,% 1,% 1,%,%,% PNGSL PRSV PNALM PNMS PNM PNGSL PRSV PNALM PNMS PNM CONCLUSION This study defined the prevalence of various internal parasites in cattle, sheep and goats within the distribution areas of the Apennine Chamois. This approach will initiate proper control strategies to minimize parasitic infections. REFERENCES. M. Asif, S. Azeem, S. Asif, and S. Nazir, Prevalence of Gastrointestinal Parasites of Sheep and Goats in and around Rawalpindi and Islamabad, Pakistan; J. Vet. Anim. Sci. (28), Vol. 1: Bashir Ahmad Lone, M.Z. Chishti, 1 1 2Fayaz Ahmad and 2Hidayatullah Tak, A Survey of Gastrointestinal Helminth Parasites of Slaughtered Sheep and Goats in Ganderbal, Kashmir; Global Veterinaria 8 (4): , 212. Bilal, M.Q, A. Hameed and T. Ahmad, Prevalence of gastrointestinal parasites in buffalo and cow calves in rural areas of toba tek singh, Pakistan; The Journal of Animal & Plant Sciences 19(2): 29, Pages: 67-7 ISSN: M. A. Raza, M. Younas and E. Schlecht, Prevalence of gastrointestinal helminths in pastoral sheep and goat flocks in the cholistan desert of Pakistan, The Journal of Animal & Plant Sciences, 24(1): 214, Page: ISSN: E. U. Edosomwan* and O. O. Shoyemi, Prevalence of gastrointestinal helminth parasites of cattle and goats slaughtered at abattoirs in Benin City, Nigeria, African Scientist Vol. 13, No. 2, June 3, 212. Cheru Telila, Birhanu Abera, Diriba Lemma and Eyob Eticha, Prevalence of gastrointestinal parasitism of cattle in East Showa Zone, Oromia Regional State, Central Ethiopia, Academic Journals, Vol. 6(2), pp , February, 214. Golo Tshering, Nedup Dorji, Prevalence of Gastrointestinal Parasites in Free Range Cattle; a Case Study in Haa District, Bhutan, Journal of Animal Health and Production. 1 (4): S. Y. Shirale, M. D. Meshram and K. P. Khillare, Prevalence of Gastrointestinal Parasites in Cattle of Western Vidarbha Region, Veterinary World, Vol.1(2): 45. V. Singh, P. Varshney, S. K. Dash2 and H. P. Lal, Prevalence of gastrointestinal parasites in sheep and goats in and around Mathura, India, doi:1.5455/vetworld J. A. Gadahi, M. J. Arshed, Q. Ali, S. B. Javaid and S. I. Shah, Prevalence of Gastrointestinal Parasites of Sheep and Goat in and around Rawalpindi and Islamabad, Pakistan, Veterinary World, Vol.2(2): Fikru Regassa, Teshale Sori, Reta Dhuguma, Yosef Kiros,Epidemiology of Gastrointestinal Parasites of Ruminants in Western Oromia, Ethiopia, Intern J Appl Res Vet Med Vol. 4, No

11 Chamois International Congress - Maiella National Park, Lama dei Peligni, Abruzzo, Central Italy - 17 th -2 th June 214 Prevalence of Coxiella, Toxoplasma and Salmonella in the ovine and caprine farms of Sirente Velino Regional Park: considerations and possibility of a holistic approach to diseases common to domestic and wild animals and humans G. Cotturone 1, E. Ruggieri 2, S. Salucci 2, P. Morini 1 1 Parco Regionale Sirente Velino 2 Istituto Zooprofilattico Sperimentale dell Abruzzo e del Molise G. Caporale, Campo Boario, 641 Teramo, Italy Introduction The present work has been carried out in the context of the the Project LIFE 9 NAT/IT/183 COORNATA, which aims at establishing a colony of Apennine chamois (Rupicapra pyrenaica ornata) within the Sirente Velino Regional Park (SVRP), as required by the National Action Plan for the Apennine Chamois (Duprè et al., 21). The SVRP was established in 1989 (LR 54/1989) and covers an area of approximately 54 km 2, which includes five Natura 2 sites (ZPS IT71113, SICIT71126, SIC IT71175, SIC IT7119, SIC IT71196). A number of endangered animal species, catalogued in the Council Directives 92/43/ EEC and 79/49/EEC (Ursus arctos marsicanus, Canis lupus, Gyps fulvus, Aquila chrysaetos, Falco biarmicus, Vipera ursinii, Rosalia alpine), currently reside in the SVRP. Recently, Lovari et al. (University of Siena) carried out a feasibility study targeted at the re-introduction of Apennine chamois in the SVRP, and a suitable area was identified on the Sirente massif (2,348 m above the sea level). The Sirente mountain is located in the central-eastern portion of the Park, has a shape of a slope long about 2 km, and is oriented NO/SE, showing a marked divergence on its side. The one exposed NE consists of beech forest and steep rocks, while the SW side has wide pastures where grazing animals are traditionally farmed. In agreement with local authorities and stakeholders, a farm-free area of protection has been defined where Apennine chamois will be re-introduced. Before the re-introduction of Apennine chamois coming from different national parks (Maiella, Monti Sibillini, Gran Sasso e Monti della Laga), specific plans were implemented in order to take a census and to evaluate the health status of domestic animals grazing in that area. Then, a specific plan was also implemented to prevent infectious disease as well as to improve the health management of domestic and syntropic animals, particularly sheep and goats (actions A12 and C6). We report herein data about that health monitoring plan, mainly focusing on the following zoonotic agents: Coxiella burnetii, Salmonella abortus ovis, Toxoplasma gondii. As known, those agents can infect humans, wild animals, and can negatively impact the economy of farms, a concept exhaustively synthesized in the sentence One World-One Health-One Medicine. Materials and Methods Monitoring activities have been carried out between July 212 and February 214, and involved a total of 4 cattle, 3, sheep and 6 goats, located in 26 farms (7 cattle and 19 sheep/goat farms). About 2% of animals residing in each farm were sampled for serological investigations (82 cattle, 672 sheep and 3 goats), a special emphasis being placed upon older subjects and reproductive disorders-affected animals. Blood samples were collected from the jugular vein using the VENOJECT system, individually identified, enclosed with a suitable form, and then referred to the Istituto Zooprofilattico Sperimentale dell Abruzzo e del Molise G. Caporale (IZSAM). Tubes were centrifuged at 3 rpm for 5 minutes, and sera were stored at +4 C (±2 C) until testing. Serological investigations were performed according to the current procedures of the IZSAM: 1. Complement fixation test (CFT) for Coxiella burnetii; 2. Slow serum agglutination test (SAT) for Salmonella abortus ovis; 3. Indirect fluorescent antibody (IFA) test for Toxoplasma gondii. During the same period of time, 12 cases of abortion occurred in sheep and goat herds. Likewise, ten fetuses were identified and referred to the IZSAM for detailed diagnostic investigations. The fetuses were kept at a +4 C (±2 C) until necropsy. In depth laboratory investigations were also carried out in order to identify the etiological agent of abortion. Serological results A total of 1,696 serological tests have been carried out, and the results are summarized in Table 1. Table 1. Serological results. Test sampled positive prevalence sampled positive prevalence sampled positive prevalence CFT Coxiella burnetii SAT Salmonella abortus ovis IFA Toxoplasma gondii Cattle Sheep Goat 72 % % % 82 % 663 % 3 % % % 1 5 5% Discussion and conclusions The health monitoring program provided interesting data about zoonotic agents, previously neglected in the territory of the SVRP. The apparent discrepancies between serological results and diagnostic investigations on aborted fetuses is likely due to some technical issues (eg. condition of samples), on one side, and to a certain distrust of shepherds, who did not allowed further diagnostic investigations, on the other. However, innovative biomolecular tools, such as real time PCR, allowed the detection of specific pathogens (eg. Coxiella burnetii). As a result of such plan, farmers were informed about the disposal of materials (fetuses, placenta, carcasses, manure), which could act as source of infection for people, domesticated and wild animals. Extensive farming (average = 2 head) is traditionally practiced in the Sirente mountain during summer (from May to October). Such familial-type herds are often managed by old farmers (5-8 years old) in an empirical way, usually without any technical support provided by veterinarians. Therefore, health issues have been addressed by a holistic approach, according to the idea that public health has to be pursued through the health of livestock and environment. Remarkably, the risk of zoonoses could be higher within the protected areas, where different categories of people (park staff, tourists, farmers, scientists) face each other and wildlife. Dairy products, produced and sold in the park, and wild animals could further contribute to transmit zoonotic infections. As a consequence, a more intense dialogue among the Park, the local veterinary authorities, the local Istituto Zooprofilattico Sperimentale and farmers seems desirable in order to plan health activities compatible with farming and exceeding the time span of this Life Project. Such an integrated approach could allow an effective control of zoonoses and, at the same time, could have a positive impact on the rural economy (eg. by quality labels for dairy products of the park). In fact, if properly managed, the maintenance of traditional sheep farming in marginal areas could play a key role to preserve the environment and biodiversity in the ecosystem of Apennines. Diagnostic investigations on aborted fetuses: results Three goat fetuses (two twins) proved to be positive for Coxiella burnetii by real time PCR. Abortion occurred during the last month of pregnancy. Serological test were carried out on the two goats which had an abortions: one was positive for Coxiella burnetii (titres 1 in 32, CFT), while both were positive for Neospora caninum (titres 1 in 32 and 1 in 64, IFA test) and negative for Toxoplasma gondii, Chlamydia abortus, Salmonella abortus ovis and border disease. Salmonella abortus ovis was isolated and identified from two twin ovine fetuses (abomasums, liver, lung, brain), while Salmonella spp. was isolated from the third fetus. Results are summarized in Table 2. Table 2. Results of diagnostic investigations on aborted fetuses* Species Number of aborted foetuses Positive for Coxiella burnetii Positive for Salmonella abortus ovis Positive for Salmonella spp Sheep Goat 3 3 Total * Laboratory investigations for Brucella spp., Chlamydia spp., Campylobacter, Bluetongue virus, Schmallenberg virus all proved to be negative. References 1. Longbottom D. & Coulter L.J. 23. Animal Chlamydioses and Zoonotic Implications. J Comp Path, 128, doi:1.153/jcpa Duprè E., Monaco A. & Pedrotti L. 21. Piano d Azione Nazionale per il camoscio appenninico (Rupicapra pyrenaica ornata). Quad Cons Natura, 1. Min. Ambiente-Ist. Naz. Fauna Selvatica. 3. Ferroglio E., Bosio F., Trisciuoglio A. & Zanet S Toxoplasma gondii in simpatriche erbivori selvatici e carnivori: epidemiologia delle infezioni nelle Alpi Occidentali. I parassiti e vettori, 7,196. doi: / Formenti N., Gaffuri A., Vicari N., Trogu T., Viganò R., Ferrari N., Paterlini F. & Lanfranchi P Diagnosi molecolare di Toxoplasma gondii da una infezione naturale alpino Chamois (Rupicapra Rupicapra r.) Dalle Alpi italiane. Bellver de Cerdanya, Spagna: II Rupicapra Symposium, Biologia, la salute, il monitoraggio e la gestione. 5. Gaffuri A., Giacometti M., Tranquillo V.M., Magnino S., Cordioli P. & Lanfranchi P. 26. Sieroprevalenza in caprioli, camosci e pecore domestiche nelle Alpi italiane centrali. J Wildl Dis, 42, Lovari S., Ferretti F. & Minder I Aggiornamento e implementazione dello studio di idoneità per l introduzione benigna nel Parco Regionale Sirente-Velino. LIFE9 NAT/IT/183 Coornata (Azione A7) Prodotto identificabile. 7. Pioz M., Loison A., Gauthier D., Gibert P., Jullien J.M., Artois M. & Gilot-Fromont E. 28. Diseases and reproductive success in a wild mammal: example in the alpine chamois. Oecologia, 155, doi: 1.17/s Moutou F. & Artois M. 21. Les mammifres sauvages reservoirs potentiels de zoonoses. Mal Infect, 31(2),

12 A TARGET TOO BIG: IMMATURE GOLDEN EAGLES ATTACKS TO ADULTS CHAMOIS Cristiani Gabriele 1, Mignone Walter 2 and Dino Scaravelli 3 1 località Colla Micheri 12/6, 1751 Andora (SV), 2 Istituto Zooprofilattico Sperimentale del Piemonte, Liguria e Val d Aosta, Sezione di Imperia 3 Dipartimento di Scienze Mediche Veterinarie, Università di Bologna, Golden eagle Aquila chrysaetos is one of the most important predator for chamois Rupicapra spp. in Alps and Pyrenees, as quoted by numerous authors, and probably also in the other parts of the species areal. Golden eagles can lift up to 5-6 kg in favorable wind and so direct predation is manly devoted to yearlings and adults are consumed as carrions. Chamois are able to engage defensive behavior especially to protect young. Mother defending offspring against eagles are known in both species and, as examples, in Ovis canadensis, Ovis gmelin, as well as in other ungulates During research on the populations of the chamois inhabits the southern slope of Marittime Alps, two cases were collected of a direct attack to adults chamois. We suppose that during the first part of its life the eagle have to try to find way to refine attack sequences and identify the suitable targets. Also the behavior to trail big prey into gorges and later scavenge on them can be considered and the sequences taken in photos can be parts of the training for the young eagle. First sequence was shot the 12/11/211 when a young eagle try to attack an isolated adult that simply ignore the bird. In the second case, at 2/11/213, the chamois respond to the attack with defensive posture and jump, showing horns, to the flying bird that leave the fight.

13 ATYPICAL COLORATION IN A SOUTH WESTERN ALPS CHAMOIS Cristiani Gabriele 1, Mignone Walter 2 and Dino Scaravelli 3 1 località Colla Micheri 12/6, 1751 Andora (SV), 2 Istituto Zooprofilattico Sperimentale del Piemonte, Liguria e Val d Aosta, Sezione di Imperia 3 Dipartimento di Scienze Mediche Veterinarie, Università di Bologna, In the Rupicapra genus is remarkable how the color pattern is common among the species and subspecies with the know differences in winter and summer appearance. Variation in color and lengths of hairs are recognized as adaptation to seasonal changes as well as in many other ungulates. Here we present a case of leucism in Rupicapra rupicapra living in the southern slope of the Marittime Alps, at the top of Imperia Province. Leucism is a phenotype with a defects in pigment cell presence in the skin and hairs, or feathers during the development. In this individuals the results is a partial or total lack of color on the body surface. The cases differ from Albino where no melanin is present at all as quoted by the red eyes in contrast with commonly colored iris in the leucistic specimens. The specimen live within its family group of 3-4 chamois were where first sighted in 211. It s the only one of the group with color variation. During the winter the group move to lower altitude with the other chamois of the area and no negative or particular interactions were noted. After a short period of thinness during last winter, now is fully recovered and moved to the tops of the pastures. All the observation do not shows any negative impact of the leucist coloration and future observation will try to study the interactions of the specimen in the groups.

14 Retrospective serological investigation on some threatening infectious agents in Apennine chamois (Rupicapra pyrenaica ornata). Alessia Di Blasio a, Maria Luisa Marenzoni a, Daria Di Sabatino b, Armando Giovannini b, Roberta Latini c, Leonardo Gentile c a Department of Veterinary Medicine, University of Perugia, via S. Costanzo 4, 6126 Perugia, Italy, b Istituto Zooprofilattico Sperimentale dell Abruzzo e del Molise G. Caporale, Campo Boario, 641 Teramo, Italy c Parco Nazionale d Abruzzo, Lazio e Molise, Via S. Lucia, 6732 Pescasseroli, AQ, Italy Introduction Apennine chamois (Rupicapra pyrenaica ornata) is a geographically isolated subspecies of Pyrenean chamois currently living only in limited areas of Central Italy. It is at present included in the red list of the International Union for Conservation of Nature (IUCN, 213) and considered a vulnerable species. Low genetic diversity, the impact of human activities, illegal hunting, straying, and trophic and spatial competition with other ungulates, like red deer and livestock, have generated this situation. Hunting of this species is prohibited. To date, four distinct populations of Apennine chamois exist. Chamois at the National Park of Abruzzo, Lazio and Molise (PNALM, n=518 chamois registered in the 29 census), represent the historical nucleus. After recent translocations, they were reintroduced at the Majella National Park (n=more than 1 chamois in 212), the National Park of Gran Sasso-Monti della Laga (n= 45 in 212) and the National park of Monti Sibillini (Raganella Pelliccioni et al. 213). An extension of the territory available for this species of chamois is needed to increase the population, but this aspect foresees a strict monitoring of the population, including infectious diseases to avoid dangerous outbreaks in this limited population because translocation of wildlife for conservation is considered a risk of disease introduction (Hartley and Lysons 21). It is very important to know if they host pathogens or if they are naïve for other pathogens, considering they often share grazing with other ungulates. However, the collection of samples is possible only when animals must be captured to move or manage them and so data of infectious diseases in this species are very limited. Aim of the study The aim of the present study was to carry out a serological retrospective survey on a sample of Apennine chamois from PNALM, captured for their handling and care, to investigate the presence of 8 of the main pathogens, which could be a possible threat for this small and isolated population of Apennine chamois because of their pathogenic role or their constant presence in wild and domestic animals. Results All captured chamois were in good clinical conditions. Serums were negative for PI-3, BTV, BHV-1, Brucella spp., Chlamydophila spp., Coxiella burnetti and Leptospira spp. The small sample size obtained in different years allowed to exclude only the presence of infections with a prevalence of 15% when 21 chamois were sampled, while in the other years when only 1 or 2 sampled chamois were investigated, this estimation was not possible. Three animals (2 females and 1 male) kept in captivity were positive at low titers (1:4 and 1:8) for pestiviruses antibodies during the year 1992 (seroprevalence 42,86%, 95% CI: 3-6%) and 5 freeranging (3 males and 2 females, seroprevalence 65,2%, 95% CI: %) with titers ranging from 1: 2 to 1:64 during 28. Materials and methods Serum samples were obtained from 12 anaesthetized Apennine chamois, during the routine marking or introduction operations carried out from 199 to 28 at the PNALM. Chamois were not necessarily captured each year. A clinical examination was performed at the time of capture. During this period the number of animals in the population ranged from 388 to 614 and it was annually determined by census. Seventy-nine samples were from free-ranging chamois, living in contact with wild and domestic animals, while the other 41 were collected from chamois kept in captivity in four distinct areas. Based on the necessities of the handling of chamois inside the park, the annual samples ranged from 1 to 21 (mean 12; median 12,5). Serum samples were stored at 2 C until use. Detection of antibodies against pestiviruses, Bovine Parainfluenza 3 virus (PI-3), Bluetongue virus (BTV), Bovine herpesvirus type 1 (BHV-1), Brucella spp., Chlamydophila spp., Coxiella burnetti and Leptospira spp. was carried out. For each infection, the most reliable serological test indicated by the World Organization for Animal Health (OIE) or a routine screening test was chosen and performed as indicated by the OIE Terrestrial Manual (OIE 213). The number of the annually sampled chamois out of the overall population of chamois at PNALM was used, when possible, to detect the presence of infection, to estimate the maximum prevalence of infection (if all samples were negative) or to estimate the expected seroprevalence with the corresponding 95% confidence interval (CI) (if positive results were present). For this analysis specific tables for sample size (Thrusfield 25) were used. Discussion The good sanitary condition of the captured animals and the results negative to PI-3, BTV, BHV-1, Brucella spp., Chlamydophila spp., Coxiella burnetti and Leptospira spp. antibodies excluded the presence of these pathogens in the population. On the other hand, the population resulted free of these etiological agents at the time of the sampling and maybe naïve and this increased the risk of outbreak, especially in a population designated to traslocations for reintroduction. The serological positivity to pestivirus confirmed the viral circulation, although its pathogenic role in this population should be further defined. Unfortunately, in the present study pestiviruses were not characterized. Considering the demonstrated extensive interspecies transmission for pestivirus, the adaptability of the virus and the detection of the infection in Pyrenean chamois 11 years before the first outbreak of disease (Gortazar et al. 27; Marco et al. 211), the pestivirus infection should be regularly monitored, especially after reports of recent outbreaks of the specific pestivirus, the Border Disease virus (BDV), which infected Pyrenean chamois (Marco et al. 211; Fernández-Sirera et al. 212a; Fernández-Sirera et al. 212b) and the presence of risk factors, like the sharing of grazing with other domestic and wild ungulates (Raganella Pelliccioni et al. 213). The present study focused on the difficulties in obtaining a good sample size to estimate the infectious status and to monitor the risk of outbreak in a small and isolated population, like the Apennine chamois. The problem with a small sample size is the reduced probability of detecting infections leading to its late discovery. Another relevant problem that makes difficult the interpretation of the results is the use of diagnostic tests not validated for wildlife species that could cause not accurate data, resulting in sensitivity and specificity that are different from those obtained for domestic animals for which the tests were originally created (Artois 21; Hartley and Lysons 211). To control infection it could be better to collect at least 5 or 11 samples/year that are able to detect infections with a seroprevalence respectively 5% or 25%, that are those frequent in the majority of infectious diseases and generally it is rare that only a few animals are infected, especially in a naïve population. The same sample size is required to find at least one positive case in the population (Thrusfield 25). Considering these results and the difficulties in collecting samples from this specific population, the sampling should be improved to obtain a better monitoring and representativeness of the sanitary status of the population of Apennine chamois in the Park. A disease screening before release could be useful to reduce the risk of disease introduction (Hartley and Lysons 211). Collection of samples from any animal of the Apennine chamois is precious for its genetic and biological value, however to monitor infectious diseases a minimum number of samples is needed to estimate the presence of infection in the population and a specific planning of sampling for infectious diseases for year and area should be foreseen.

15 RED DEER AND APENNINE CHAMOIS: A DIFFICULT COEXISTENCE Francesco Ferretti 1,*, Marcello Corazza 2, Isabelle Minder 1, Ilaria Campana 1, Venusta Pietrocini 1, Claudia Brunetti 1, Davide Scornavacca 1, Natalia Troiani 2,Carlo Ferrari 2 & Sandro Lovari 1 1 Research Unit of Behavoural Ecology, Ethology and Wildlife Management, Dept. of Life Sciences, University of Siena. Via P.A. Mattioli 4, 531, Siena, Italy. 2 Dept. of Evolutioary Biology, University of Bologna. Via Irnerio 42, 4126, Bologna, Italy. * BACKGROUND and AIMS Information is scarce on mechanisms of competition between wild ungulate species. Since the early 2s, a c. 5% decline in abundance of Apennine chamois Rupicapra pyrenaica ornata has occurred in the core area of their distribution range, in Abruzzo, Lazio and Molise Nat. Park (ALMNP), because of a great winter mortality of kids, whereas numbers of red deer Cervus elaphus (reintroduced in ) have greatly increased (1; 2). The potential competition between deer and chamois was evaluated in terms of (i) grassland changes in respect to the time when deer were absent (3); (ii) effects of ecological overlap of these ungulates on quality of grassland and foraging efficiency of nursing female chamois. Area A Area B Area C Study areas: Fig. 1. Study period: July-early November Fig. 1 Study areas: AREA A: Val di Rose (c. 3 ha, 17-2 m a.s.l.) high deer density AREA B: Mt. Amaro (c. 18 ha, m a.s.l.) medium deer density AREA C: Mt. Meta (c. 3 ha, m a.s.l.) red deer absent We assessed: PASTURE COMPOSITION/QUALITY (veg. surveys; GLMs) DIET OVERLAP OF DEER AND CHAMOIS (micro-histological analyses of pellets) EFFECTS OF DEER DENSITY ON PASTURE (veg. surveys; GLMMs) CHAMOIS FORAGING EFFICIENCY (behavioural observations and GLMMs) RESULTS ) AREA A: GRASSLAND CHANGES AND DEER/CHAMOIS OVERLAP (1) With respect to (3): signif. decrease of 11 out of 12 most grazed species by chamois in the 198 s (freq. occurrence and/or cover); signif. sharp increase of unpalatable species (frequency/cover). Seasonal diet overlap (Pianka index): always >.85. Deer pellet groups in 76-88% plots, seasonally used by chamois ) AREAS A-B-C: EFFECTS OF DEER ON: % COVER % VOLUME Trampling increase July - October AREA A AREA B AREA C Leguminosae in diet AREA A AREA B AREA C SUMMER AUTUMN Fig. 2 Trampling cover (n =568 surveys; 71 plots) was signif. greater in Areas A-B than in C (no-deer area). Fig. 3 In each season/veg. type (n=113 obs. bouts, 1 min/bout): feeding intensity of female chamois signif. the greatest in Area C (no deer); food searching rate signif. the greatest in Area A (highest deer density). CONCLUSIONS N BITES TO GRASS / MIN N STEPS / MIN Feeding intensity AREA A AREA B AREA C Food searching AREA A AREA B AREA C Fig. 4 The volume of Leguminosae (the best food items for chamois) in diet (n = pellets/season): did not vary seasonally in Area C (no deer); decreased significantly from summer to autumn in Areas A-B. Mother s condition and availability of high-quality forage in the warm season influence early growth and winter survival of offspring (e.g. 4; 5; 6). Grazing/trampling by red deer affected food availability, diet quality and foraging efficiency of nursing female chamois. In turn, winter survival of chamois kids would be affected, leading to negative effects on pop. structure and dynamics (1; 2). The reintroduction of a non-threatened species - potentially competing with a threatened one - may not be always advisable. REFERENCES 1. Lovari S, Ferretti F, Corazza M, Minder I, Troiani N, Ferrari C, Saddi S (in press). Anim Conserv. 2. Latini R, Gentile L, Asprea A, Pagliaroli D, Argenio A, Di Pirro V (211). ALMNP Agency, unpublished report. 3. Ferrari C, Rossi G, Cavani C. (1988). Z Säugetierkd. 53: Clutton-Brock TH, Albon SD, Guinness FE (1986) Anim Behav 34: Côté SD, Festa-Bianchet M (21). Oecologia127: ACKNOWLEDGEMENTS Funding: Abruzzo, Lazio and Molise Nat. Park Agency (ALMNP), integrated by LIFE 9NAT/IT/183 COORNATA; Italian Ministry of University and Research (PRIN project n. 21P7LFW4). We are grateful to G. Rossi, D. Febbo; ALMNP personnel; A. Saddi for help with data collection.

16 LAZY GLUTTON MALES, WISE PICKY FEMALES: THE FORAGING BEHAVIOUR OF APENNINE CHAMOIS Francesco Ferretti 1,*, Alessia Costa 1, Marcello Corazza 2, Gloria Cesaretti 1 & Sandro Lovari 1 1 Research Unit of Behavoural Ecology, Ethology and Wildlife Management, Dept. of Life Sciences, University of Siena. Via P.A. Mattioli 4, 531, Siena, Italy. 2 Dept. of Evolutioary Biology, University of Bologna. Via Irnerio 42, 4126, Bologna, Italy. * BACKGROUND and AIMS In dimorphic ungulates, females are usually more selective foragers, show greater bite rates and spend more time foraging than males (e.g. 1; 2; 3; but see 4). This pattern could be explained through (i) body size-related differences of metabolic rates, (ii) energy requirements determined by reproduction. Chamois Rupicapra spp. show a sexual size dimorphism (SSD) limited only to several months before and during the rut (5; 6). Thus, in summer-early autumn, females and males should have comparable, great energy requirements: the former must gain weight before rut, the latter nurse their offspring. We evaluated sexual differences of foraging behaviour in Apennine chamois Rupicapra pyrenaica ornata (7). If sexual differences of foraging behaviour depend on SSD, we expect that, in summer-autumn, female chamois (a) show greater bite and step rates, (b) spend more time feeding, (c) select higher-quality food patches than males. Study period: July-early November We assessed: BITE RATE / STEP RATE (behavioural observationsand GLMMs) SELECTION OF VEGETATION (behavioural observationsand GLMMs) ACTIVITY BUDGET (behavioural observations and G-tests) Study area: upper Val di Rose (Abruzzo, Lazio and Molise Nat. Park, c m a.s.l., right) and its location in Italy (left). RESULTS (a) BITE RATE / STEP RATE The bite rate was signif. greater in males than in females; the step rate showed the opposite pattern. (b) SELECTION OF VEGETATION Both sexes selected nutritious patches (veg. with Trifolium thalii), with no signif. sexual differences. (c) ACTIVITY BUDGET The two sexes spent a comparable amount of time feeding. Males lay down a longer time than females. CONCLUSIONS (1) In summer-autumn, both sexes met their energy requirements by selecting nutritious (clover) patches, with different tactics: MALES lower selectivity, faster intake; FEMALES greater selectivity, slower intake. (2) Not only body size-related energy requirements, but especially those related to life history strategies and reproductive costs play an important role in determining sexual differences of foraging behaviour. (3) Great food intake rate of males, in the warm season behavioural adaptation leading to evolutionary transition from year round monomorphism to permanent dimorphism, through seasonal-dimorphism? REFERENCES 1. Gross JE, Demment MW, Alkon PU, Kotzman M (1995). Funct Ecol 9: Ruckstuhl KE (1998) Anim Behav 56: Ruckstuhl KE, Festa-Bianchet M, Jorgenson JT (23) Behav Ecol Sociobiol 54: Pérez-Barbería FJ, Robertson E, Soriguer R, Aldezabal A, Mendizabal M, Pérez-Fernández E (27). Ecol Monogr 77: Garel M, Loison A, Jullien JM, Dubray D, Maillard D, Gaillard JM (29) J Mammal 9: Rughetti M, Festa-Bianchet M (211) J Zool 284: ACKNOWLEDGEMENTS Funding: Abruzzo, Lazio and Molise Nat. Park Agency (ALMNP); Italian Ministry of University and Research (PRIN project n. 21P7LFW4). We are grateful to G. Rossi, D. Febbo; ALMNP personnel; V. Pietrocini, A. Saddi, N. Troiani for help with data collection; C. Ferrari for supervising vegetation analyses; L. Corlatti, J. Pérez-Barberìa and K. Ruckstuhl for comments.

17 Serosurvey for selected pathogens in sheep and cattle in the areas of presence of the Apennine Chamois (Rupicapra pyrenaica ornata) in five protected area in Central Apennine in Italy L. Gentile1, G. Cotturone2, U. Di Nicola3, F. Morandi4, S. Angelucci5, M. Innocenti3,5, V. Di Pirro1, A. Argenio1, E. Carosi1, R. Latini1, S. Gavaudan6, F. Barchiesi6, P. Morini2, F. Striglioni3, A. Rossetti4, M. Scacchia7, M. Tittarelli7 1Abruzzo, 4Sibillini Lazio and Molise National Park, 2Sirente-Velino Regional Park, 3Gran Sasso and Mountain of Laga National Park, Mountain National Park, 5Majella National Park, 6IZS Umbria and Marche, 7IZS Abruzzo and Molise G. Caporale. INTRODUCTION From 211 to 214, during the Life + Coornata Project, in the activities related to the action C.6, antibody seroprevalence in n cattle and n sheep and goats were investigated in five protected area in Central Apennine in Italy: Abruzzo, Lazio and Molise National Park, Majella National Park, Gran Sasso and Mountain of Laga National Park, Sibillini Mountain National Park and Sirente-Velino Regional Park, where the population of Appennine Chamois (Rupicapra pyrenaica ornata) are expanding and increasingly in contact with livestock. MATERIALS AND METHODS Study area Our study area was five protected areas in Central Apennine. 1. Abruzzo, Lazio and Molise National Park (PNALM): n hectares; 2. Majella National Park (PNM): n ; 3. Gran Sasso and Mountain of Laga National Park (PNGSL): n hectares; 4. Sirente-Velino Regional Park (PRSV): n hectares; 5. Sibillini Mountain National Park (PNMS): n hectares. Animal sampling We investigated antibody seroprevalence against: Neospora caninum (n. 595 sera samples), Chlamydia psittaci ovis (n. 4.95), Coxiella burnetii (n ), Salmonella abortus ovis (n ), Toxoplasma (n ), Border disease (n ), Mycoplasma agalactiae (n. 2.71), Mycobacterium paratuberculosis (n ), Borrelia spp. (n. 487), Contagious Ecthyma (n. 145), Bovine Viral Diarrhea (n. 715), Infectious bovine rhinotracheitis (n. 245), Parainfluenza virus type 3 (n. 67), Anaplasma phagocitophila (n. 44), and Leptospira spp. (n. 31). RESULTS The Serologic prevalence against different pathogens in the areas of presence of the apennine chamois, are summarized in the graphics n. 1 (in sheep and goats) and n. 2 (in cattle). GRAPHIC n. 2 - Serologic prevalence of selected infectious diseases in cattle GRAPHIC n. 1 - Serologic prevalence of selected infectious diseases in sheep and goats 9,% 6,% 8,% 7,% 5,% 6,% 4,% 5,% 4,% 3,% 3,% 2,% 2,% 1,% 1,%,%,% The number of positive sera of selected infectious diseases in sheep and goats against different pathogens for only protected areas, are summarized the followed graphics. Positive sera of selected infectious diseases in sheep and goats in PNGSL Positive sera of selected infectious diseases in sheep and goats in PNALM Sera Positive sera of selected infectious diseases in sheep and goats in PNMS Positive sera of selected infectious diseases in sheep and goats in PRSV Sera Positive sera Sera Positive sera Positive sera of selected infectious diseases in sheep and goats in PNM Sera Positive sera Sera Positive sera The number of positive sera of selected infectious diseases in cattle against different pathogens for only protected areas, are summarized the followed graphics. Positive sera of selected infectious diseases in cattle in PNGSL Positive sera of selected infectious diseases in cattle in PRSV Positive sera of selected infectious diseases in cattle in PNALM Sera Positive sera Sera Positive sera Positive sera of selected infectious diseases in cattle in PNMS Sera Positive sera Sera Positive sera of selected infectious diseases in cattle in PNM Sera Positive sera Positive sera CONCLUSION To our knowledge, this is the first study reporting prevalence and risk factors associated with certain infectious agents in the areas of presence of apennine chamois. Moreover, our results suggest to continue and study in deep epidemiology of some diseases in the areas of presence and spread of apennine chamois, not only throughout a serological monitoring but also by research directed to the agents in particular in ewes and cows after parturition and/or abortion. REFERENCES. M. Travnicek, D. Kovacova, P. Zubricky, L. CislaKova, Serosurvey of sheep and goats to Chlamydia psittaci in Slovakia during the yers , Vet. Med. Czech, 46, 21 (11 12): Maria Silvia Gennero, Stefania Bergagna, Marco Pasino, Antonio Barbaro, Angelo Romano, Anna Trisciuoglio and Ezio Ferroglio, Neospora caninum serological survey in cattle from the Piedmont Region (northwestern Italy), Epidémiol. et santé anim., 27, 51, Ntafis, V., Xylouri, E., Diakou, A., Sotirakoglou, K., Kritikos, I., Georgakilas, E. and Menegatos, I., Serological survey of antibodies against toxoplasma gondii in organic sheep and goat farms in Greece, ISAH-27 Tartu, Estonia. By M. Lundervold, E.J. Milner-Gulland, C.J. O'Callaghan, C. Hamblin, A. Corteyn4 and A.P. Macmillan, A Serological Survey of Ruminant Livestock in Kazakhstan During Post-Soviet Transitions in Farming and Disease Control, Acta vet. scand. 24, 45, L.F. Pita Gondim, H.V. Barbosa Jr., C.H.A. Ribeiro Filho, H. Saeki, Serological survey of antibodies to Toxoplasma gondii in goats, sheep, cattle and water buffaloes in Bahia State, Brazil, Veterinary Parasitology 82 (1999) Francisco Ruiz-Fons, Ianire Astobiza, Jesús F Barandika, Ana Hurtado, Raquel Atxaerandio, Ramón A Juste, Ana L García-Pérez, Seroepidemiological study of Q fever in domestic ruminants in semi-extensive grazing systems, Ruiz-Fons et al. BMC Veterinary Research 21, 6:3. By M. Lundervold, E.J. Milner-Gulland, C.J. O'Callaghan, C. Hamblin, A. Corteyn and A.P. Macmillan, A Serological Survey of Ruminant Livestock in Kazakhstan During Post-Soviet Transitions in Farming and Disease Control, Acta vet. scand. 24, 45, Eva Aisser Botres Ajaj, Maab. I. AL- Farwachi, Serosurvey of Leptospira interrogans Serovars hardjo and pomona in Cattle in Nineveh Province, Iraq, Animal Health Prod and Hyg (213) 2(1) : V. Fridriksdóttir, L. L. Nesse and R. Gudding, Seroepidemiological studies of Borrelia burgdorferi infection in sheep in Norway, J. Clin. Microbiol. 1992, 3(5):1271. Ana C. Coelho, Maria L. Pinto, Adosinda M. Coelho, Alfredo Aires and Jorge Rodrigues, A seroepidemiological survey of Mycobacterium avium subsp. paratuberculosis in sheep from North of Portugal, Pesq. Vet. Bras. 3(11):93-98, novembro 21.

18 The effects of pulmonary deficiencies on a vulnerable Apennine Chamois population require a cautionary immobilization protocol Gentile L., Asprea A., Pagliaroli D., Argenio A., Di Pirro V. & Latini R. Servizio Scientifico e Veterinario del Parco Nazionale d Abruzzo, Lazio e Molise The Apennine chamois (Rupicapra pyrenaica ornata) Endemic subspecies which survives in some massifs of Central Italy; IUCN conservation status down-ranked from Endangered to Vulnerable; major threats still occurring and associated with: limited population size; limited number of subpopulations; low genetic variation; competition with other ungulates. The Apennine chamois found in the Abruzzo, Lazio and Molise National Park constitutes the last remaining autochthonous population and it has an especially high conservation value. STUDY AREA: Abruzzo, Lazio and Molise National Park (PNALM) About 55 Km 2 ; rich community of wild ungulates (red deer Cervus elaphus, roe deer Capreolus capreolus, wild boar Sus scrofa, other than Apennine chamois); broad presence of domestic livestock (357 cows, 2191 goats, 74 sheep and several horses). Apennine Chamois were captured in two of the most representative areas of the PNALM: Val di Rose and La Meta. Val di Rose La Meta Since 25-26, the PNALM population has shown: decreasing growth rate (λ)( (running average, s.d. <> ) decreasing kids' survival rate in the first year (running average, s.d. <>.15-.3) high level of parasites, mostly pulmonar strongyles (N = 6) (N = 138) (N = 45) Strongyles occurrence significantly higher than that previously recorded (χ( 2, P <.1). In addition: after over twenty years with no mortality during capture operations, in 26 two individuals died on the same day; in 28 and 29 another two individuals died during anesthesia; 84.2% of 17 chamois retrieved between 21 and 213, including 3 juveniles, had poor pulmonary conditions caused by high infestation of strongyles. Possible density-dependent processes caused by several factors (e.g. high intraspecific local-density, reduced carrying capacity due to changes in production and species diversity, and to interspecific competition for grazing pastures). Clinical conditions, broadly occurring in the PNALM population, that: cannot be predicted from a visual assessment of the subject s external conditions; represent a potential risk in case of immobilization and manipulation. AIM Xilazine-Ketamine (XK) or Medetomidine-Ketamine (MK)? Which of the two protocols usually applied by PNALM staff is more appropriate to reduce the risks for the animals? Two periods: and Study made within the ongoing project LIFE9 NAT/IT/183 Coornata. MATERIAL AND METHODS 14 chamois immobilizations since 199: 83 using Xilazine-Ketamine (XK); 21 using Medetomidine-Ketamine (MK). Before 26, XK was used 57 times and MK 11; since 26, XK was used 26 times and MK 1 times. The parameters considered were the following: induction time (IT, in minutes); recovery time (RT, in minutes); age, sex and weight of the animal; dosage (mg) of first drug injection; heart rate (HR); respiratory rate (RR); first rectal temperature recorded after recovery (T). RESULTS 1) No correlation between induction time and injection site for both the protocols. No correlation with body weight, sex, age and first dosage all data were treated together. None of the mortality events occurred (3 for each protocol) was related to a specific anesthesiological aspect. HR and T did not correlate with the time since immobilization for both the protocols. 2) Parameter MK (mean ± sd) XK (mean ± sd) Mann-Whitney U test IT prior ± ± 3. n.s. IT after ± ± 1.3 U = 124.5, P <.5 RT 2.8 ± ± 3.8 n.s. HR 51.2 ± ± 15. n.s. RR 69.9 ± ± 23.4 n.s RR showed a significant negative correlation with the time since immobilization for both protocols, but the relationship for MK was stronger (r = -.47 vs. r = -.28, P <.5). A less variable decreasing RR is usually associated with a deep anesthesia and consequent decreasing stress level or adaptation to stress. CONCLUSION According to our results, we recommend the use of MK protocol, since it yields a minor induction time, thus allowing to get quicker immobilization; would also allow the measures required to be started sooner, thus minimizing the respiratory problems that may occur during the anesthesia; seems to be more effective to obtain a deeper anesthesia because of the higher regularity of some anesthesiological parameters (RR in particular). Acknolowdgements We are grateful to D. Russo for helping us with English. LIFE9 NAT/IT/183 Development of coordinated protection measures for Apennine Chamois (Rupicapra pyrenaica ornata) COORNATA Chamois International Congress Lama dei Peligni (AQ) - June, 17-19, 214

19 Monitoring of a reintroduced population of Cantabrian Chamois in the Aramo Mountain Range (North of Spain) Pablo González-Quirós* Jaime Marcos** Óscar Rodríguez*** Carlos Nores**** *Consultora BIOGESTION. Cimadevilla 15, Esc. B-2º D; 333-Oviedo (Spain) ** Dirección General de Recursos Naturales del Principado de Asturias; 3371-Oviedo (Spain) ***IREC Universidad de Castilla La Mancha; 1371-Ciudad Real (Spain) ****INDUROT Universidad de Oviedo. Campus de Mieres; 336-Mieres (Spain) Study area The Aramo Mountain Range is a protected natural area (Protected Landscape), close to the Cantabrian chamois Rupicapra pyrenaica parva core area in the Cantabrian mountain range, where the chamois was present up the beginning of the XX th century Calcareous massif of 54, ha (6. ha of suitable habitat) a maximum altitude of 1,786 m and steep rocky slopes. (Fig. 1) THE REINTRODUCTION Planning (24) FIGURE 1.- The top of the Aramo Mountain Range covered with snow shows its best habitat for chamois. Extinction historical study (23): Where, when & why? The probable date of extinction was first decades of the XX th century due to overhunting (Fig. 2) Habitat quality (23): Where & how much? Matching suitability and availability (Fig. 3) Captures (24): Where & how many? Somiedo Natural Park (25-3 km; <28 chamois) 75 individuals, netting was done from September to March Executing (27-212) Capture device: What & how? Vertical nets were placed in forest edges (3-5 m long) 7-1 handlers (every 5 m), 6-8 beaters, 3-5 blockers (mean 2.4 captures/day) (Fig. 4 A) Animal welfare: What? Chamois were released in the capture site if > 6 specimens were caught (8 individuals), immobilization, blinding, anesthetising, sanitary control, backpacking on foot (3-4 min.) (Fig. 4 B), individual transport boxes (1x36x76 cm) (Fig. 4 C) by car. No particular diseases Mortality: When, why & how many? During the captures 4.2%, 12.5% after the release (first weeks) FIGURE 2.- Present range of chamois in the Cantabrian mountain range. Dots show historical records during XIXth century. The Aramo area is shown in the circle and the translocation is indicated with an arrow. Table I. Main parameters of catches in Somiedo Natural Park, during the years Catch area Year Month No. days PARQUE NATURAL DE SOMIEDO Sept. - March Sept. - Oct. Sept. - Oct. Sept. - Dec No. people net No. beaters Chamois catch (ind./day) 34 (2.6) 15 (3.) 1 (3.3) 16 (2.7) Sexratio 1: 2.3 Mortality catch % 2.9 (1M) 1: 2. 1: 5. 1: (1M) 6.3 (1F) Mortality post release % 11.8 (1M+3F) 8.3 (1F) 2. (2F) 12.5 (1M+1F) FIGURE 3.- Four types of habitat suitability models. The Aramo area is shown within the ellipse Sept. - March (2.4) 1: (1M) 16. (1M+2H) TOTAL (mean) 1 (2.4) 1: Release A B Release sites: Where? Three different sites. West, East and South (3-4 km) ( individuals) Age structure: What? Age and sex structure was similar to that of the source population (Fig. 5) Tagging: How & how many? Ear tags (all); encoded colour collars (adults >2 years); radio-tracking emitters (7 VHF, 9 GPS/GSM) (Fig. 6) FIGURE 4.- Three stages of handling animals: (A) Capture device; (B) transporting and (C) releasing. Monitoring Materials: What? GPS collars gave 1,5-8,5 fixes/year, visual monitoring 3-4 day/week (Fig. 7) Habitat: Where? Using mainly areas >5% slope Spreading: How many & how far? 9 individuals > 7 km; 3 returned to the capture area (17-22 km away) (Fig. 8) Demography: How? Mean natality.22; mean mortality+emigration (observed emigration 2-3 ind./year [Fig. 6] ).115; λ (considering only no released animals) 1,46 Age (years) C Table II. Population trend and demographic parameters of the reintroduction individuals Released Total Young Natality Mortality+emigration λ t+1/t ,16 FIGURE 5.- Population structure (sex and age) of released chamois. FIGURE 7.- Movements of three GPS radio-tagged chamois during 29 (from 23/4-7/5). Red and yellow dots are fixes of two females. The blue ones belong to a male. ACKNOWLEDGEMENTS FIGURA 6- Released chamois with a conventional radio tracking collar and an ear tag. FIGURE 8- Longest dispersions from the releasing sites outside the study area. Blue stars and lilac dots belong to two chamois that returned to their original locations. This project was inspired by the Regional Government of the Principado de Asturias, and funded by an EU project LEADER de la Montaña Central de Asturias. It was also supported by the hunting societies of Morcín, Quirós, Lena and the Instituto de Investigación en Recursos Naturales, sponsored by private funders as Repsol, Gas Natural, BWM and TRAGSEGA. Field work collaborators: Valentín Morán, Sergio Solano, Laureano Prieto, Javier Antuña, Marco García Gala, Alfonso González, Raúl Ríos, Jorge Martí, Álvaro Oleaga, Miguel Prieto, Ana Balseiro, Alberto Spi, Rafael Alba, José Luis García, José Armenteros, the park rangers of the Principado de Asturias and many others.

20 SPATIAL AND TEMPORAL PHYLOGENY OF BORDER DISEASE VIRUS IN PYRENEAN CHAMOIS Luzzago C. 1, Ebranati E. 2, Lanfranchi P. 1, Cabezón O. 3,4, Lavín S. 3, Rosell R. 4, Rossi L. 5, Zehender G. 2, Marco I. 3 1 Dept. Veterinary Science and Public Health, University of Milan, Italy; : 2 Dept. Clinical Sciences Luigi Sacco, University of Milan, Italy; 3 Servei d'ecopatologia de Fauna Salvatge, Dept. Medicina i Cirurgia Animals, Universitat Autònoma de Barcelona, Bellaterra, Spain; 4 Centre de Recerca en Sanitat Animal (CReSA). UAB-IRTA, Campus de la Universitat Autònoma de Barcelona, Bellaterra, Spain; 5 Dept. Veterinary Sciences, University of Torino, Grugliasco, Italy BACKGROUND Border disease virus (BDV) (Genus Pestivirus, Family Flaviviridae) affects a wide range of ruminants worldwide, mainly domestic sheep and goat. Since 21 several outbreaks of disease associated to BDV infection have been described in Pyrenean chamois (Rupicapra pyrenaica pyrenaica) in Spain, France and Andorra (fig. 1) (1, 2, 3). These outbreaks have decimated several Pyrenean chamois populations, with mortalities ranging from 4% to 85%. The infection has become endemic in the Central and Eastern Pyrenees. After the severe BDV outbreaks, different epidemiological scenarios have appeared in the Pyrenees, some of which are having a negative impact on host population dynamics (4). AIM The aim was to clarify the origin and dispersion of the Pyrenean chamois BDV genetic variant by reconstructing the spatial and temporal dynamics of BDV 5 UTR sequences of Pyrenean chamois. MATERIALS AND METHODS Ten novel BDV sequences of Pyrenean chamois and 41 retrieved from public databases were analyzed. Sheep BDV sequences (n=43) from Spain and France were also retrieved from public databases. A phylogenetic analysis was performed using a Bayesian Markov chain Monte Carlo (MCMC) method implemented in the BEAST v.1.74 package (5). Statistical support for specific clades was obtained by calculating the posterior probability of each monophyletic clade. The trees were summarised in a target tree, choosing the tree with the maximum product of posterior probabilities (maximum clade credibility) after a 1% burn-in. Figure 1 Distribution areas of Pyrenean chamois Figure 2 Eastern Pyrenees Western Pyrenees Central Pyrenees RESULTS AND DISCUSSION The maximum clade credibility tree, summarizing all of the trees obtained during the MCMC (fig. 2), showed a main clade supported by posterior probabilities of 1, corresponding to the Pyrenean chamois phylogenetic group. The chamois clade originated from sheep BDV showing a relatively recent emergence (mean estimate 1992). There were also some significant sub-clades among chamois sequences clustering different geographical areas in Pyrenees. CONCLUSION Our data suggest that Pyrenean chamois phylogenetic group originated from sheep BDV genotype 4, generating a founder effect due to intraspecies spread and spatial dispersion, still going on such as Western direction. ACKNOWLEDGMENTS Work funded by the PRIN Grant prot. 21P7LFW4 Genomics and host-pathogen interactions: a model study in the One-Health perspective from the Italian Ministry of Education, Scientific Research and Health. REFERENCES 1. Marco et al., 29.Border Disease Virus among Chamois, Spain. Emerging Infectious Diseases 15, 448-5; 2. Pioz et al., 27. Transmission of a pestivirus infection in a population of Pyrenean chamois. Veterinary Microbiology 119, 19 3; 3. Fernandez-Sirera et al., 212a. Surveillance of BDV in wild ungulates and an outbreak in Pyrenean chamois (Rupicapra pyrenaica pyrenaica) in Andorra. Journal of Wildlife Diseases, 48, ; 4. Fernandez-Sirera et al., 212b Two Different Epidemiological Scenarios of Border Disease in the Populations of Pyrenean chamois (Rupicapra p. pyrenaica) after the First Disease Outbreaks. PLOSONE 7, e5131; 5.Drummond, A.J., et al., 212. Bayesian phylogenetics with BEAUti and the BEAST 1.7. Molecular Biology and Evolution 29,

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