Serologic Evidence for Selected Infectious Diseases in Marsican Brown Bears (Ursus arctos marsicanus) in Italy (2004–09) Source: Journal of Wildlife Diseases, 51(1):209-213. Published By: Wildlife Disease Association URL: http://www.bioone.org/doi/full/10.7589/2014-01-021
BioOne (www.bioone.org) is a nonprofit, online aggregation of core research in the biological, ecological, and environmental sciences. BioOne provides a sustainable online platform for over 170 journals and books published by nonprofit societies, associations, museums, institutions, and presses. Your use of this PDF, the BioOne Web site, and all posted and associated content indicates your acceptance of BioOne’s Terms of Use, available at www.bioone.org/page/terms_of_use. Usage of BioOne content is strictly limited to personal, educational, and non-commercial use. Commercial inquiries or rights and permissions requests should be directed to the individual publisher as copyright holder.
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SHORT COMMUNICATIONS DOI: 10.7589/2014-01-021
Journal of Wildlife Diseases, 51(1), 2015, pp. 209–213 # Wildlife Disease Association 2015
Serologic Evidence for Selected Infectious Diseases in Marsican Brown Bears (Ursus arctos marsicanus) in Italy (2004–09) Cristina Esmeralda Di Francesco, 1,4 Leonardo Gentile, 2 Vincenza Di Pirro, 2 Lara Ladiana, 1 Silvia Tagliabue,3 and Fulvio Marsilio1 1Veterinary Faculty, University of Teramo, P.za A. Moro, 45, 64100 Teramo, Italy; 2Abruzzo Lazio and Molise National Park, Viale Santa Lucia, 67032 Pescasseroli (AQ), Italy; 3 National Reference Centre for Leptospirosis, Istituto Zooprofilattico della Lombardia e dell’Emilia Romagna, Via Bianchi, 9, 25124 Brescia, Italy; 4Corresponding author (email: [email protected])
per la Protezione e la Ricerca Ambientale [ISPRA] 2001), edited by the Italian Ministry of the Environment for the Protection of the Territory and the Sea, recognizes as priority the collection of data on the health status of the Marsican brown bear with particular reference to infectious diseases that can be transmitted to the bear from other wildlife or domestic species living in the same habitat (ISPRA 2011). However, little information is available about pathogens affecting the Marsican brown bear population and the impact on its health status (Marsilio et al. 1997). To improve knowledge of the distribution of infectious diseases in the Marsican brown bear population, we surveyed for antibodies against canine distemper virus (CDV), canine adenovirus type 2 (CAV2), canine parvovirus type 2 (CPV-2), Brucella spp., and eight Leptospira interrogans sensu lato serovars in free-ranging bears captured within PNALM during 2004–09. Thirty sera were collected from 22 bears, 1–15 yr old, captured by Aldrich foot snares and chemically immobilized with three mixtures of drugs (Marsilio et al. 1997). Sera were heated at 56 C for 30 min and tested by virus neutralization (VN) for CDV and CAV-2 (Lennette et al. 1988), by hemagglutination inhibition for CPV-2 (Carmichael et al. 1980), by rapid serum agglutination for Brucella abortus strain 99 (Weybridge) and Brucella melitensis, and by microagglutination test (MAT) for eight L. interrogans sensu lato serovars (Bratislava; Ballum; Canicola;
ABSTRACT: We tested 30 serum samples collected during 2004–09 from 22 free-ranging Marsican brown bears (Ursus arctos marsicanus) in the National Park of Abruzzo, Lazio, and Molise, Italy, for antibodies against canine distemper virus (CDV), canine adenovirus type 2 (CAV-2), canine parvovirus type 2 (CPV-2), Brucella spp., and eight Leptospira interrogans sensu lato serovars. Antibody to CDV was detected in 11 samples (37%); only two bears (10%) had detectable CAV-2 and Brucella spp. antibodies; three bears were positive for L. interrogans serovar Bratislava; and one sample had antibody against L. interrogans serovar Copenhageni. All samples were positive for CPV-2 antibody. The CPV-2 antibody titers varied from 1:640 to 1:10,240, suggesting that transmission was still active. Fifty percent of bears were positive for antibody to two or more pathogens. Our results highlight the need to consider infectious diseases as a potential risk for Marsican brown bear conservation. Key words: Brown bear, infectious diseases, serology, Ursus arctos marsicanus.
The Marsican brown bear (Ursus arctos marsicanus) is a subspecies of the brown bear (Ursus arctos; Altobello 1921; Randi et al. 1994) listed in Appendix I of the Convention on International Trade in Endangered Species of Wild Fauna and Flora (CITES 1979) and included in the International Union for Conservation of Nature (IUCN) Red List (McLellan et al. 2008). The population of Marsican brown bears is very small (40–50 individuals; Gervasi et al. 2008) and is restricted to the National Park of Abruzzo, Lazio, and Molise (PNALM) in the Apennine Mountains of Italy (41u489N, 13u479E). The Action Plan for the Conservation of the Marsican brown bear (Istituto Superiore 209
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TABLE 1. Titers for antibodies to canine distemper virus (CDV), canine adenovirus type 2 (CAV-2), canine parvovirus type 2 (CPV-2), Brucella spp., and Leptospira interrogans sensu lato serovars in samples collected from 22 free-ranging Marsican brown bears (Ursus arctos marsicanus) living in the National Park of Abruzzo, Lazio, and Molise, Italy. Some bears were captured multiple times during capture activities in 2004–09. These captures are identified by a letter (a, b, or c) after the bear identification (ID) number. Dashes (—) indicate negative test results. Sampling year Age (years)
Bear ID
Sexa
1-a 1-b 1-c 2 3-a 3-b 4 5 6 7-a 7-b 8 9 10 11 12 13 14-a
F F F F M M M F F M M F F M F M M M
2004 2007 2008 2004 2005 2006 2005 2005 2006 2006 2008 2006 2006 2006 2006 2007 2007 2007
14-b 15-a 15-b 16 17-a 17-b 18 19-a 19-b 20 21 22
M M M M M M F F F F F M
2009 2008 2009 2008 2008 2009 2008 2008 2009 2009 2009 2007
a
CDV
CAV-2
8–10 10–12 12–15 10–12 10–12 10–12 8–10 6–7 8–10 8–10 10–12 4–5 12–15 7–8 8–10 10–12 3–4 4–5
1:16 1:16 1:32 — 1:64 1:32 1:8 — 1:8 1:8 — — — — — 1:32 — —
— — — — 1:32 1:16 — — — — — — — — — — — —
6–7 6–7 7–8 5–6 4–5 5–6 1–2 12–15 12–15 1–2 4–5 4–5
1:32 — — — — — — — — — — $1:256
— — 1:16 — — — — — — — — —
Brucella abortus/ B. melitensis
Leptospira interrogans sensu lato serovar (titer)
1:2,560 1:2,560 1:2,560 1:2,560 1:1,280 1:1,280 1:2,560 1:2,560 1:1,280 1:2,560 1:640 1:2,560 1:2,560 1:2,560 1:2,560 1:2,560 1:2,560 1:1,280
— — — — Positive Positive Positive — — — — — — — — — — —
1:1,280 1:2,560 1:2,560 1:2,560 1:5,120 1:5,120 1:5,120 1:5,120 1:5,120 1:5,120 1:5,120 1:10,240
— — — — — — — — — — — —
— — — — — — — — — — — — — — — — — Bratislava (1:1,600)/ Copenagheni (1:200) Bratislava (1:400) — — Bratislava (1:100) — — — Bratislava (1:100) — — — —
CPV-2
F 5 female; M 5 male.
Grippotyphosa; Copenhageni; Pomona; Hardjo; Tarassovi) (Cole et al. 1973). Serologic results are summarized in Table 1; Figures 1 and 2 show the geographic and temporal distribution of positive bears. Eleven sera collected from eight bears (8/22, 36%) were positive for CDV antibody with titers up to $1:256, and three samples from two animals (2/ 22, 9%) had antibody to CAV-2. Antibody titers to CDV are consistent with studies in the same area by Marsilio et al. (1997), suggesting that the source of infection may be identified in domestic or stray
dogs (Canis lupus familiaris) living in PNALM along with other wild carnivores susceptible to CDV infection such as wolves (Canis lupus) and foxes (Vulpes vulpes). An outbreak of canine distemper occurred during 2013 in dogs and wolves in PNALM with several animals found dead or rescued alive with severe clinical signs attributed to CDV (Di Sabatino et al. 2014). Thus, the prospect of the spread of canine distemper in Marsican bears is not completely unfounded, even if the effects of the infection on the population are unknown and the clinical
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FIGURE 1. Geographic distribution of the National Park of Abruzzo, Lazio, and Molise (Italy) sampling sites where captured Marsican brown bears (Ursus arctos marsicanus) were found positive for antibodies to canine distemper virus (CDV), canine adenovirus type 2 (CAV-2), canine parvovirus type 2, Brucella spp., and Leptospira interrogans sensu lato serovars. &5Sites with bears positive for antibody to CDV; m5Bears positive for antibodies to CAV2; ¤5Bears positive for antibodies to Brucella spp.; 5Bears positive for antibody to Leptospira interrogans.
disease has not been described in this species. Our finding of antibody to CAV-2 is the first evidence of virus circulation in the Marsican brown bear population. Infection probably results from occasional contact with other infected domestic or wild canids. The VN test we used does not distinguish between CAV-1 and CAV-2 due to cross-reactivity (Green 2006), so it is not possible to demonstrate which adenovirus is circulating in the Marsican bear population. Susceptibility of the bears to canine adenoviruses has been documented only for CAV-1 in captive black bears (Ursus americanus), which show typical clinical signs and postmortem lesions of infectious hepatitis (Collins et al. 1984). In wildlife, however, CAV-2 circulation is reported in foxes, wolves, and coyotes (Canis latrans) but not in bears (Gerhold et al. 2007; Balboni et al. 2013).
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FIGURE 2. Temporal prevalence of antibodies (percentage of bears with antibody) to canine distemper virus (CDV), canine adenovirus type 2 (CAV-2), canine parvovirus type 2 (CPV-2), Brucella spp., and Leptospira interrogans sensu lato serovars in Marsican brown bears (Ursus arctos marsicanus) in the National Park of Abruzzo, Lazio, and Molise, Italy, 2004–09.
Antibodies against CPV-2 were detected in all examined sera, with titers ranging from 1:640 to 1:10,240, suggesting that the infection is endemic in the population without evident mortality. It is possible that a host-adapted strain of CPV-2 is circulating in wildlife that has achieved a stable population immunity. In a survey of 83 environmental stool specimens from Marsican brown bears, by electron microscopy, Potena et al. (2005) detected parvovirus-like particles with hemoagglutinant properties, but no other information about the virus was obtained. The presence of CPV-2 is documented for other wild species such as wolves and foxes from Italy, Spain, and Portugal (Battilani et al. 2001; Sobrino et al. 2008; Santos et al. 2009). The stability of the virus in the environment should allow interspecies transmission and the maintenance of a sylvatic cycle of infection. Two bears were antibody positive to both Brucella antigens and four were positive by MAT for Bratislava serovar; one also had antibodies to L. interrogans serovar Copenhageni. The antibodies to Brucella spp. show that the bears have been infected even if the prevalence in the population and the source of infection are
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unknown. In the PNALM area, brucellosis continues to occur in livestock, and the Officially Free status for B. abortus and B. melitensis has not been achieved. Sheep (Ovis aries), which are the most-common livestock in the area, are likely the source of B. melitensis for bears, especially during transhumance activities, while the antibodies to B. abortus are due to antigenic similarity between lipopolysaccharides of the two bacteria (Corbel et al. 1984). Cross-reaction among Brucella species also occurs for Brucella suis, which was recently isolated from a wild boar found dead in the Abruzzo region (De Massis et al. 2012). Therefore, the hypothesis that the bears are also exposed to B. suis cannot be ruled out. The Leptospira results represent the first attempt to evaluate the occurrence of this pathogen in Marsican brown bears; their serum was reactive against the serovar Bratislava, which is deposited widely in the environment by wild and domestic animals including hedgehogs (Erinaceus europaeus), foxes, dogs, and pigs (Sus scrofa) (Ellis 1989; Bolin and Cassels 1990; Hartskeerl and Terpstra 1996; Scanziani et al. 2002). The antibodies against the serovar Copenhageni may represent a cross-contamination between the Icterohaemorrhagiae and Australis serogroups. Investigators in Croatia detected antibody in .35% of sera collected from brown bear populations, showing a strong correlation with the serovars detected in other small mammals of the region (Modric and Huber 1993; Slavica et al. 2010). In Marsican brown bears the low number of antibody-positive animals suggests that the impact of leptospirosis on the health of this population is negligible. In conclusion, the small size of the Marsican brown bear population, its total isolation, and the densely populated habitat of the PNALM represent major threats despite strict application of the conservation plan. Infectious diseases could be an additional risk. Vaccination
of domestic carnivores should be implemented in the National Park and in the surrounding territories to avoid epidemic outbreaks that could involve critically endangered species. LITERATURE CITED Altobello G. 1921. Mammiferi. IV. Carnivori. Fauna dell’Abruzzo e del Molise. Colitti, Campobasso, Italia, 61 pp. Balboni A, Verin R, Morandi F, Poli A, Prosperi S, Battilani M. 2013. Molecular epidemiology of canine adenovirus type 1 and type 2 in freeranging red foxes (Vulpes vulpes) in Italy. Vet Microbiol 162:551–557. Battilani M, Scagliarini A, Tisato E, Turilli C, Jacoboni I, Casadio R, Prosperi S. 2001. Analysis of canine parvovirus sequences from wolves and dogs isolated in Italy. J Gen Virol 82:1555–1560. Bolin C, Cassels JA. 1990. Isolation of Leptospira interrogans serovar Bratislava from stillborn and weak pigs in Iowa. J Am Vet Med Assoc 196:1601– 1604. Carmichael LE, Joubert JC, Pollock RVH. 1980. Hemagglutination by canine parvovirus: Serologic studies and diagnostic applications. Am J Vet Res 41:784–792. Cole JR, Sulzer CR, Pursell AR. 1973. Improved micro-technique for the leptospiral microscopic agglutination test. Appl Microbiol 28:976–980. Collins JE, Leslie P, Johnson D, Nelson D, Peden W, Boswell R, Draayer H. 1984. Epizootic of adenovirus infection in American black bears. J Am Vet Med Assoc 185:1430–1432. Convention on International Trade in Endangered Species of Wild Fauna and Flora (CITES). 1979. Appendices I, II and III, http://www.cites.org/ eng/app/appendices.php. Accessed January 2014. Corbel MJ, Stuart FA, Brewer RA. 1984. Observations on serological cross-reactions between smooth Brucella species and organisms of other genera. Dev Biol Stand 56:341–348. De Massis F, Di Provvido A, Di Sabatino D, Di Francesco D, Zilli K, Ancora M, Tittarelli M. 2012. Isolation of Brucella suis biovar 2 from a wild boar in the Abruzzo Region of Italy. Vet Ital 48:397–404. Di Sabatino D, Lorusso A, Di Francesco CE, Gentile L, Di Pirro V, Bellacicco AL, Giovannini A, Di Francesco G, Marruchella G, Marsilio F, et al. 2014. Arctic lineage-canine distemper virus as a cause of death in Apennine wolves (Canis lupus) in Italy. PLoS One 9:e82356. Ellis WA. 1989. Leptospira Australis infection in pigs. Pig Vet J 22:83–89. Gerhold RW, Allison AB, Temple DL, Chamberlain MJ, Strait KR, Keel MK. 2007. Infectious canine
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hepatitis in a gray fox (Urocyon cinereoargenteus). J Wildl Dis 43:734–736. Gervasi V, Ciucci P, Boulanger JB, Posillico M, Sulli C, Focardi S, Randi E, Boitani L. 2008. Preliminary estimate of the Apennine brown bear population size based on hair-snag sampling and multiple data source mark–recapture Huggins models. Ursus 19:105–121. Greene C. 2006. Infectious diseases of the dog and cat, 3rd Ed. Elsevier Saunders, St. Louis, Missouri, 1,424 pp. Hartskeerl RA, Terpstra WJ. 1996. Leptospirosis in wild animals. Vet Quart 18:149–150. Istituto Superiore per la Protezione e la Ricerca Ambientale (ISPRA). 2011. National action plan for the conservation of Marsican brown bear. Quad Cons Natura 37:1–59. Lennette EH, Halonen P, Murphy FA. 1988. Laboratory diagnosis of infectious diseases: Principles and practice, Vol. II. Springer-Verlag Publisher, Berlin, Germany, 1,071 pp. Marsilio F, Tiscar PG, Gentile L, Roth HU, Boscagli G, Tempesta M, Gatti A. 1997. Serologic survey for selected viral pathogens in brown bears from Italy. J Wildl Dis 33:86–89. McLellan BN, Servheen C, Huber D (International Union for Conservation of Nature [IUCN] SSC Bear Specialist Group). 2008. Ursus arctos. The IUCN Red List of Threatened Species. Version 2014.2, www.iucnredlist.org. Accessed January 2014. Modric´ Z, Huber D. 1993. Serological survey for leptospirosis in European brown bears (Ursus arctos) in Croatia. J Wildl Dis 29:608–611.
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Potena G, Sammarone L, Battisti A, Filippone I, Gentile L, Brugnola L, Petrella A, Opramolla G. 2005. Health status of the Marsican brown bear population and other related species. Project LIFE99 NAT/IT/006244 ‘‘Brown bear conservation in Central Apennine’’ Ministry of Agriculture and Forestry, http://www3.corpoforestale.it/ flex/cm/pages/ServeBLOB.php/L/IT/IDPagina/ 1151. Accessed January 2014. Randi R, Gentile L, Boscagli G, Huber D, Roth HU. 1994. Mitochondrial DNA sequence divergence among some west European brown bear (Ursus arctos L.) populations. Lessons for conservation. Heredity 73:480–489. Santos N, Almendra C, Tavares L. 2009. Serologic survey for canine distemper virus and canine parvovirus in free-ranging wild carnivores from Portugal. J Wildl Dis 45:221–226. Scanziani E, Origgi F, Giusti AM, Iacchia G, Vasino A, Pirovano G, Scarpa P, Tagliabue S. 2002. Serological survey of leptospiral infection in kennelled dogs in Italy. J Small Anim Pract 43:154–157. Slavica A, Konjevic´ D, Huber D, Milas Z, Turk N, Sindicic M, Severin K, Dezdek D, Masek T. 2010. Serologic evidence of Leptospira spp. serovars in brown bears (Ursus arctos) from Croatia. J Wildl Dis 46:251–256. Sobrino R, Arnal MC, Luco DF, Gorta´zar C. 2008. Prevalence of antibodies against canine distemper virus and canine parvovirus among foxes and wolves from Spain. Vet Microbiol 126:251–256. Submitted for publication 28 January 2014. Accepted 1 April 2014.
BioOne (www.bioone.org) is a nonprofit, online aggregation of core research in the biological, ecological, and environmental sciences. BioOne provides a sustainable online platform for over 170 journals and books published by nonprofit societies, associations, museums, institutions, and presses. Your use of this PDF, the BioOne Web site, and all posted and associated content indicates your acceptance of BioOne’s Terms of Use, available at www.bioone.org/page/terms_of_use. Usage of BioOne content is strictly limited to personal, educational, and non-commercial use. Commercial inquiries or rights and permissions requests should be directed to the individual publisher as copyright holder.
BioOne sees sustainable scholarly publishing as an inherently collaborative enterprise connecting authors, nonprofit publishers, academic institutions, research libraries, and research funders in the common goal of maximizing access to critical research.
SHORT COMMUNICATIONS DOI: 10.7589/2014-01-021
Journal of Wildlife Diseases, 51(1), 2015, pp. 209–213 # Wildlife Disease Association 2015
Serologic Evidence for Selected Infectious Diseases in Marsican Brown Bears (Ursus arctos marsicanus) in Italy (2004–09) Cristina Esmeralda Di Francesco, 1,4 Leonardo Gentile, 2 Vincenza Di Pirro, 2 Lara Ladiana, 1 Silvia Tagliabue,3 and Fulvio Marsilio1 1Veterinary Faculty, University of Teramo, P.za A. Moro, 45, 64100 Teramo, Italy; 2Abruzzo Lazio and Molise National Park, Viale Santa Lucia, 67032 Pescasseroli (AQ), Italy; 3 National Reference Centre for Leptospirosis, Istituto Zooprofilattico della Lombardia e dell’Emilia Romagna, Via Bianchi, 9, 25124 Brescia, Italy; 4Corresponding author (email: [email protected])
per la Protezione e la Ricerca Ambientale [ISPRA] 2001), edited by the Italian Ministry of the Environment for the Protection of the Territory and the Sea, recognizes as priority the collection of data on the health status of the Marsican brown bear with particular reference to infectious diseases that can be transmitted to the bear from other wildlife or domestic species living in the same habitat (ISPRA 2011). However, little information is available about pathogens affecting the Marsican brown bear population and the impact on its health status (Marsilio et al. 1997). To improve knowledge of the distribution of infectious diseases in the Marsican brown bear population, we surveyed for antibodies against canine distemper virus (CDV), canine adenovirus type 2 (CAV2), canine parvovirus type 2 (CPV-2), Brucella spp., and eight Leptospira interrogans sensu lato serovars in free-ranging bears captured within PNALM during 2004–09. Thirty sera were collected from 22 bears, 1–15 yr old, captured by Aldrich foot snares and chemically immobilized with three mixtures of drugs (Marsilio et al. 1997). Sera were heated at 56 C for 30 min and tested by virus neutralization (VN) for CDV and CAV-2 (Lennette et al. 1988), by hemagglutination inhibition for CPV-2 (Carmichael et al. 1980), by rapid serum agglutination for Brucella abortus strain 99 (Weybridge) and Brucella melitensis, and by microagglutination test (MAT) for eight L. interrogans sensu lato serovars (Bratislava; Ballum; Canicola;
ABSTRACT: We tested 30 serum samples collected during 2004–09 from 22 free-ranging Marsican brown bears (Ursus arctos marsicanus) in the National Park of Abruzzo, Lazio, and Molise, Italy, for antibodies against canine distemper virus (CDV), canine adenovirus type 2 (CAV-2), canine parvovirus type 2 (CPV-2), Brucella spp., and eight Leptospira interrogans sensu lato serovars. Antibody to CDV was detected in 11 samples (37%); only two bears (10%) had detectable CAV-2 and Brucella spp. antibodies; three bears were positive for L. interrogans serovar Bratislava; and one sample had antibody against L. interrogans serovar Copenhageni. All samples were positive for CPV-2 antibody. The CPV-2 antibody titers varied from 1:640 to 1:10,240, suggesting that transmission was still active. Fifty percent of bears were positive for antibody to two or more pathogens. Our results highlight the need to consider infectious diseases as a potential risk for Marsican brown bear conservation. Key words: Brown bear, infectious diseases, serology, Ursus arctos marsicanus.
The Marsican brown bear (Ursus arctos marsicanus) is a subspecies of the brown bear (Ursus arctos; Altobello 1921; Randi et al. 1994) listed in Appendix I of the Convention on International Trade in Endangered Species of Wild Fauna and Flora (CITES 1979) and included in the International Union for Conservation of Nature (IUCN) Red List (McLellan et al. 2008). The population of Marsican brown bears is very small (40–50 individuals; Gervasi et al. 2008) and is restricted to the National Park of Abruzzo, Lazio, and Molise (PNALM) in the Apennine Mountains of Italy (41u489N, 13u479E). The Action Plan for the Conservation of the Marsican brown bear (Istituto Superiore 209
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TABLE 1. Titers for antibodies to canine distemper virus (CDV), canine adenovirus type 2 (CAV-2), canine parvovirus type 2 (CPV-2), Brucella spp., and Leptospira interrogans sensu lato serovars in samples collected from 22 free-ranging Marsican brown bears (Ursus arctos marsicanus) living in the National Park of Abruzzo, Lazio, and Molise, Italy. Some bears were captured multiple times during capture activities in 2004–09. These captures are identified by a letter (a, b, or c) after the bear identification (ID) number. Dashes (—) indicate negative test results. Sampling year Age (years)
Bear ID
Sexa
1-a 1-b 1-c 2 3-a 3-b 4 5 6 7-a 7-b 8 9 10 11 12 13 14-a
F F F F M M M F F M M F F M F M M M
2004 2007 2008 2004 2005 2006 2005 2005 2006 2006 2008 2006 2006 2006 2006 2007 2007 2007
14-b 15-a 15-b 16 17-a 17-b 18 19-a 19-b 20 21 22
M M M M M M F F F F F M
2009 2008 2009 2008 2008 2009 2008 2008 2009 2009 2009 2007
a
CDV
CAV-2
8–10 10–12 12–15 10–12 10–12 10–12 8–10 6–7 8–10 8–10 10–12 4–5 12–15 7–8 8–10 10–12 3–4 4–5
1:16 1:16 1:32 — 1:64 1:32 1:8 — 1:8 1:8 — — — — — 1:32 — —
— — — — 1:32 1:16 — — — — — — — — — — — —
6–7 6–7 7–8 5–6 4–5 5–6 1–2 12–15 12–15 1–2 4–5 4–5
1:32 — — — — — — — — — — $1:256
— — 1:16 — — — — — — — — —
Brucella abortus/ B. melitensis
Leptospira interrogans sensu lato serovar (titer)
1:2,560 1:2,560 1:2,560 1:2,560 1:1,280 1:1,280 1:2,560 1:2,560 1:1,280 1:2,560 1:640 1:2,560 1:2,560 1:2,560 1:2,560 1:2,560 1:2,560 1:1,280
— — — — Positive Positive Positive — — — — — — — — — — —
1:1,280 1:2,560 1:2,560 1:2,560 1:5,120 1:5,120 1:5,120 1:5,120 1:5,120 1:5,120 1:5,120 1:10,240
— — — — — — — — — — — —
— — — — — — — — — — — — — — — — — Bratislava (1:1,600)/ Copenagheni (1:200) Bratislava (1:400) — — Bratislava (1:100) — — — Bratislava (1:100) — — — —
CPV-2
F 5 female; M 5 male.
Grippotyphosa; Copenhageni; Pomona; Hardjo; Tarassovi) (Cole et al. 1973). Serologic results are summarized in Table 1; Figures 1 and 2 show the geographic and temporal distribution of positive bears. Eleven sera collected from eight bears (8/22, 36%) were positive for CDV antibody with titers up to $1:256, and three samples from two animals (2/ 22, 9%) had antibody to CAV-2. Antibody titers to CDV are consistent with studies in the same area by Marsilio et al. (1997), suggesting that the source of infection may be identified in domestic or stray
dogs (Canis lupus familiaris) living in PNALM along with other wild carnivores susceptible to CDV infection such as wolves (Canis lupus) and foxes (Vulpes vulpes). An outbreak of canine distemper occurred during 2013 in dogs and wolves in PNALM with several animals found dead or rescued alive with severe clinical signs attributed to CDV (Di Sabatino et al. 2014). Thus, the prospect of the spread of canine distemper in Marsican bears is not completely unfounded, even if the effects of the infection on the population are unknown and the clinical
SHORT COMMUNICATIONS
FIGURE 1. Geographic distribution of the National Park of Abruzzo, Lazio, and Molise (Italy) sampling sites where captured Marsican brown bears (Ursus arctos marsicanus) were found positive for antibodies to canine distemper virus (CDV), canine adenovirus type 2 (CAV-2), canine parvovirus type 2, Brucella spp., and Leptospira interrogans sensu lato serovars. &5Sites with bears positive for antibody to CDV; m5Bears positive for antibodies to CAV2; ¤5Bears positive for antibodies to Brucella spp.; 5Bears positive for antibody to Leptospira interrogans.
disease has not been described in this species. Our finding of antibody to CAV-2 is the first evidence of virus circulation in the Marsican brown bear population. Infection probably results from occasional contact with other infected domestic or wild canids. The VN test we used does not distinguish between CAV-1 and CAV-2 due to cross-reactivity (Green 2006), so it is not possible to demonstrate which adenovirus is circulating in the Marsican bear population. Susceptibility of the bears to canine adenoviruses has been documented only for CAV-1 in captive black bears (Ursus americanus), which show typical clinical signs and postmortem lesions of infectious hepatitis (Collins et al. 1984). In wildlife, however, CAV-2 circulation is reported in foxes, wolves, and coyotes (Canis latrans) but not in bears (Gerhold et al. 2007; Balboni et al. 2013).
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FIGURE 2. Temporal prevalence of antibodies (percentage of bears with antibody) to canine distemper virus (CDV), canine adenovirus type 2 (CAV-2), canine parvovirus type 2 (CPV-2), Brucella spp., and Leptospira interrogans sensu lato serovars in Marsican brown bears (Ursus arctos marsicanus) in the National Park of Abruzzo, Lazio, and Molise, Italy, 2004–09.
Antibodies against CPV-2 were detected in all examined sera, with titers ranging from 1:640 to 1:10,240, suggesting that the infection is endemic in the population without evident mortality. It is possible that a host-adapted strain of CPV-2 is circulating in wildlife that has achieved a stable population immunity. In a survey of 83 environmental stool specimens from Marsican brown bears, by electron microscopy, Potena et al. (2005) detected parvovirus-like particles with hemoagglutinant properties, but no other information about the virus was obtained. The presence of CPV-2 is documented for other wild species such as wolves and foxes from Italy, Spain, and Portugal (Battilani et al. 2001; Sobrino et al. 2008; Santos et al. 2009). The stability of the virus in the environment should allow interspecies transmission and the maintenance of a sylvatic cycle of infection. Two bears were antibody positive to both Brucella antigens and four were positive by MAT for Bratislava serovar; one also had antibodies to L. interrogans serovar Copenhageni. The antibodies to Brucella spp. show that the bears have been infected even if the prevalence in the population and the source of infection are
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unknown. In the PNALM area, brucellosis continues to occur in livestock, and the Officially Free status for B. abortus and B. melitensis has not been achieved. Sheep (Ovis aries), which are the most-common livestock in the area, are likely the source of B. melitensis for bears, especially during transhumance activities, while the antibodies to B. abortus are due to antigenic similarity between lipopolysaccharides of the two bacteria (Corbel et al. 1984). Cross-reaction among Brucella species also occurs for Brucella suis, which was recently isolated from a wild boar found dead in the Abruzzo region (De Massis et al. 2012). Therefore, the hypothesis that the bears are also exposed to B. suis cannot be ruled out. The Leptospira results represent the first attempt to evaluate the occurrence of this pathogen in Marsican brown bears; their serum was reactive against the serovar Bratislava, which is deposited widely in the environment by wild and domestic animals including hedgehogs (Erinaceus europaeus), foxes, dogs, and pigs (Sus scrofa) (Ellis 1989; Bolin and Cassels 1990; Hartskeerl and Terpstra 1996; Scanziani et al. 2002). The antibodies against the serovar Copenhageni may represent a cross-contamination between the Icterohaemorrhagiae and Australis serogroups. Investigators in Croatia detected antibody in .35% of sera collected from brown bear populations, showing a strong correlation with the serovars detected in other small mammals of the region (Modric and Huber 1993; Slavica et al. 2010). In Marsican brown bears the low number of antibody-positive animals suggests that the impact of leptospirosis on the health of this population is negligible. In conclusion, the small size of the Marsican brown bear population, its total isolation, and the densely populated habitat of the PNALM represent major threats despite strict application of the conservation plan. Infectious diseases could be an additional risk. Vaccination
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