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Epidemiological survey of zoonotic pathogens in feral pigeons (Columba livia var. domestica) and sympatric zoo species in Southern Spain
Accepted Manuscript Title: Epidemiological survey of zoonotic pathogens in feral pigeons (Columba livia var. domestica) and sympatric zoo species in Southern Spain Author: David Cano-Terriza Rafael Guerra Silvie Lecollinet Marta Cerd`a-Cu´ellar Oscar Cabez´on Sonia Almer´ıa Ignacio Garc´ıa-Bocanegra PII: DOI: Reference:
S0147-9571(15)00078-8 http://dx.doi.org/doi:10.1016/j.cimid.2015.10.003 CIMID 1030
To appear in: Received date: Revised date: Accepted date:
22-6-2015 23-9-2015 17-10-2015
Please cite this article as: Cano-Terriza D, Guerra R, Lecollinet S, Cerdgravea-Cu´ellar M, Cabez´on O, Almer´ia S, Garc´ia-Bocanegra I, Epidemiological survey of zoonotic pathogens in feral pigeons (Columba livia var. domestica) and sympatric zoo species in Southern Spain., Comparative Immunology, Microbiology and Infectious Diseases (2015), http://dx.doi.org/10.1016/j.cimid.2015.10.003 This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and review of the resulting proof before it is published in its final form. Please note that during the production process errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain.
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Epidemiological survey of zoonotic pathogens in feral pigeons (Columba livia var.
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domestica) and sympatric zoo species in Southern Spain.
3 Authors
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David Cano-Terrizaa, Rafael Guerrab, Silvie Lecollinetc, Marta Cerdà-Cuéllard, Oscar
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Cabezóne, Sonia Almeríad,f, Ignacio García-Bocanegraa *
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a
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Excellence International Campus (ceiA3), Campus de Rabanales, 14071, Córdoba,
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Dept. de Sanidad Animal, Facultad de Veterinaria, Universidad de Córdoba-Agrifood
Spain.
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b
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Córdoba, Spain.
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c
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INRA, ANSES, ENVA, Maisons-Alfort, F-94703, France.
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Agroalimentàries (IRTA), Campus de la Universitat Autònoma de Barcelona,
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Bellaterra, Barcelona, Spain.
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e
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Animals, Universitat Autònoma de Barcelona, 08193, Bellaterra, Barcelona, Spain.
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f
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de Barcelona, 08193, Bellaterra, Barcelona, Spain.
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Parque Zoológico Municipal de Córdoba (PZMC), Avenida Linneo s/n, 14071,
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ANSES, Laboratoire de Santé Animale de Maisons-Alfort, UMR 1161 Virologie,
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Centre de Recerca en Sanitat Animal (CReSA) - Institut de Recerca i Tecnologia
Servei d'Ecopatologia de Fauna Salvatge (SEFaS), Dept. de Medicina i Cirugia
Dept. de Sanitat i d’Anatomia Animals, Facultat de Veterinària, Universitat Autònoma
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* Corresponding author at: Departamento de Sanidad Animal (Universidad de
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Córdoba). Edificio Sanidad Animal 2ª planta, Campus Universitario Rabanales, 14071,
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Córdoba, Spain. Tel: +34 95 721 8725; fax: +34 95 721 8727.
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E-mail address: [email protected] (Ignacio García-Bocanegra) 1 Page 1 of 21
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Abstract A cross-sectional study was carried out to determine the prevalence of
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pathogenic zoonotic agents (flaviviruses, avian influenza viruses (AIVs), Salmonella
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spp. and Toxoplasma gondii) in feral pigeons and sympatric zoo animals from Córdoba
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(Southern Spain) between 2013 and 2014. Antibodies against flaviviruses were
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detected in 7.8% out of 142 (CI95%: 3.7-11.8) pigeons, and 8.2% of 49 (CI95%: 0.9-
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15.4) of zoo animals tested. Antibodies with specificity against West Nile virus
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(WNV) and Usutu virus (USUV) were confirmed both in pigeons and in zoo birds.
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Even though seropositivity to AIVs was not detected in any of the analyzed pigeons,
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17.9% of 28 (CI95%: 3.7-32.0) zoo birds tested showed positive results. Salmonella spp.
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was not isolated in any of 152 faecal samples collected from pigeons, while 6.8% of 44
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zoo animals were positive. Antibodies against T. gondii were found in 9.2% of 142
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(CI95%: 4.8-13.6) feral pigeons and 26.9% of 108 (CI95%: 19.6-34.1) zoo animals. This
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is the first study on flaviviruses and T. gondii in feral pigeons and captive zoo species
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in Spain. Antibodies against WNV and USUV detected in non-migratory pigeons and
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captive zoo animals indicate local circulation of these emerging pathogens in the study
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area. T. gondii was widespread in species analyzed. This finding could be of
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importance for Public Health and Conservation of endangered species present in zoo
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parks. Pigeons and zoo animals may be included as sentinel species for monitoring
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zoonotic pathogens in urban areas.
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Keywords: Avian influenza viruses; Captive zoo animals; Feral pigeons; Flaviviruses;
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Salmonella spp.; Toxoplasma gondii.
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Highlights
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•
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Spain •
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•
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these species
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Circulation of Flaviviruses, AIV, Salmonella spp. and T. gondii were detected in
The results obtained could have relevance for Public Health and Conservation
Pigeons and zoo animals may be included as
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We report prevalence of zoonotic pathogens in feral pigeons and zoo animals in
sentinel species for monitoring zoonotic pathogens in urban areas
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1. Introduction Feral pigeon (Columba livia var. domestica; Columbidae family) is one of the
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bird species most frequently found in urban and peri-urban areas. During the last
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decades, their populations have increased exponentially in different countries, reaching
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densities higher than 2000 birds/km2 in many European cities [1,2]. Favorable
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environmental conditions and abundant food, which imply loosing the breeding season,
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and the absence of predators are the main factors implicated in the high increase of their
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populations [3]. Feral pigeon is considered a pest species in many urban areas due to the
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destruction architectonic and urban heritage, damage to agriculture and risk of disease
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transmission to other sympatric species, including humans. In this sense, control
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measures including surgical or chemical sterilization, removal of eggs, physical
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repellents, use of birds of prey or capturing and controlled elimination are frequently
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implemented in urban areas [2,4].
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Zoological parks are a very favorable habitat for pigeons, where they are found
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in higher densities than in urban areas. High populations imply significant economic
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losses associated to the consumption of food intended for captive zoo animals.
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Furthermore, the risk of disease transmission from pigeons to zoo sympatric species can
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be of concern for Animal Health and Conservation. In this sense, the importance of zoos
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for surveillance of zoonotic and emerging diseases has been previously suggested [5].
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Even though feral pigeons are considered reservoirs of zoonotic diseases
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including
Newcastle
disease,
histoplasmosis,
ornithosis,
salmonellosis,
or
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cryptococcosis [6,7], information about zoonotic diseases in this species in Spain is yet
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very limited. High prevalence of Chlamydophila psittaci infection was found in central
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(Madrid: 52.6%) and southwest (Murcia: 35.9%) areas [8,9]. Prevalence of 4 Page 4 of 21
Campylobacter spp. infection ranged between 1.1% and 69.1% in Barcelona and
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Madrid, respectively [10,9]. Also, 1.5% of Salmonella spp. and 0.25% of Yersinia
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intermedia infections was detected in pigeons from Barcelona city [10]. In addition,
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70.3% of pigeons analyzed in Alicante (southwest Spain) showed Cryptococcus
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neoformans infection [11].
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The aim of this study was to determine the prevalence of selected zoonotic
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pathogens (flaviviruses, avian influenza viruses (AIVs), Salmonella spp. and T. gondii)
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in feral pigeons, and to assess the risk of transmission to sympatric zoological species in
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the Córdoba Municipal Zoo Park (CMZP, Southern Spain).
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2. Materials and Methods
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2.1 Study design and sampling
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A total of 152 feral pigeons were captured in the CMZP (Southern Spain)
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between November 2013 and May 2014. Captures were performed within the course of
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a pest control program undertaken by the local authorities following European
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guidelines (UNE 171210: 2008). Animals were handled and sampled according to
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regulations of the Regional Government of Andalusia. Blood samples were taken from the brachial vein, after those animals were
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humanely euthanized by overdose sodium pentobarbital (Dolethal®, Vetoquinol
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Laboratories, Lure, France) (0.5 ml/kg) as recommended for birds [12]. Samples
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were placed into sterile tubes without anticoagulant and centrifuged at 3000 rpm for 10
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min. Sera were stored at -20ºC until analysis. All individuals were necropsied and
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inspected for the presence of macroscopic lesions. Pigeons were classified as juveniles
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(< 8 months) and adult (> 8 months) according to previously established criteria [13]. 5 Page 5 of 21
Sera from 108 zoo animals belonging to 34 species (Table 1), which share habitat with
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the sampled pigeons, were also collected. In addition, digestive content (2-3 gr) from all
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pigeons and 44 fresh faecal samples from 35 different zoo species were collected for
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Salmonella spp. isolation.
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2.2 Laboratory analysis
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Sera from 142 pigeons and 49 captive zoo animals were tested for antibodies
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against one epitope of the preMembrane-Envelope (prM-E) protein common to
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flaviviruses of the Japanese Encephalitis antigenic complex (JEV) using a commercial
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blocking ELISA (bELISA 10.WNV.K3 INGEZIM West Nile COMPAC®, Ingenasa,
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Madrid, Spain). ELISA-positive sera were confirmed by virus neutralization test (VNT)
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for the detection of specific neutralizing antibodies against West Nile virus (WNV; Is98
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strain), Usutu virus (USUV; It12 strain) and Meaban virus (MBV; Brest ART707
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strain). VNTs were performed as previously described [14,15]. Samples that showed
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neutralization and absence of cytopathic effect at dilutions ≥ 10 for WNV and USUV
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and ≥ 20 for MBV, were considered positive. Interpretation of results was based on
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comparison of VNT titers obtained in parallel against the 3 flaviviruses. The
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neutralizing immune response observed was considered specific when VNT titers for a
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given virus was ≥ 4-fold higher than titers obtained for the other viruses. Samples
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showing VNT titers differences ≤ 2-fold between the viruses examined were considered
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positive for flavivirus but not conclusive for any specific virus.
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The presence of antibodies against the nucleoprotein of AIVs type A was
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determined using a commercial (bELISA) (1.0.FLU.K3 INGEZIM INFLUENZA A®,
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Ingenasa, Madrid, Spain) according to the manufacturer´s recommendations. A total of
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148 pigeons and 28 zoo birds were analyzed. 6 Page 6 of 21
Faecal droppings and faecal samples were processed as described in Annex D of
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the Spanish standard UNE-EN ISO 6579:2002 for Salmonella isolation. Salmonella-
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presumptive colonies were tested with the Mucap (Biolife, Milano, Italy) and indole
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tests. Salmonella serotyping was carried out according to the Kauffman-White scheme
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[16] by the Agri-food laboratory of Departament d’Agricultura, Ramaderia, Pesca,
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Alimentació i Medi Natural (Generalitat de Catalunya, Spain).
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Sera from 142 feral pigeons and 108 zoo animals, including samples from 28
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carnivores, 43 ungulates, 35 birds and 2 monkeys were examined to detect antibodies
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against T. gondii using the modified agglutination test (MAT) as previously described
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[17]. Serum was tested at 1:25, 1:50, 1:100 and 1:500 dilutions and both positive and
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negative controls were included in all tests. Sera with titers ≥ 1:25 were considered
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positive.
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2.3 Statistical analysis
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The prevalence of the different pathogens was estimated from the ratio of
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positive to the total number of samples tested, with the exact binomial confidence
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intervals of 95% (CI95%). Correlation between estimated prevalences and independent
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variables (species, sex and age) were analyzed by means of a Pearson’s chi-square test
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or, when there were less than six observations per category, by the Fisher´s exact test. The
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differences between variables were analyzed by Tukey tests. Values with P < 0.05 were
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considered as statistically significant. Statistical analyses were performed using SPSS
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22.0 (Statistical Package for Social Sciences (SPSS) Inc., Chicago, IL, USA).
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3. Results Antibodies against flavivirus of the JEV serocomplex were found in 11 of 142
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pigeons (7.8%; CI95%: 3.7-11.8) tested by bELISA. No statistically significant
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differences were observed among sex or age. Six of the 11 positive samples were
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confirmed by VNT (Table 2). Specific antibodies against WNV were detected in three
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pigeons (3/142: 2.1%; CI95%: 0.0-4.3), while USUV infection was also confirmed in
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three individuals (3/142: 2.1%; CI95%: 0.0-4.3). No MBV-specific antibodies were
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detected in the pigeon sera tested.
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Seroprevalence of flaviviruses in zoo animals was 8.2% (4/49; CI95%: 0.9-15.4).
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Presence of antibodies was found in ostrich (Struthio camelos), white stork (Ciconia
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ciconia), emperor goose (Chen canagica) and black wildebeest (Connochaetes gnou)
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(Table 2). Specific antibodies against WNV (1:40) and USUV (1:20) were confirmed in
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white stork and ostrich, respectively. Moreover, the seropositive emperor goose found
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positive in bELISA presented neutralizing antibodies against WNV (1:80), USUV
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(1:40) and MBV (1:20), which are indicative of at least two independent flavivirus
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infections (by a flavivirus belonging to the JEV serocomplex like WNV or USUV and
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tick-borne flavivirus like MBV).
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Antibodies against AIV were not found in any of the 148 pigeons tested (0.0%;
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CI95%: 0.0-1.9). Seropositivity was observed in five out of 28 (17.9%; CI95%: 3.7-32.0)
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sera from captive zoo birds, including mallard (Anas platyrhynchos), emperor goose,
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African sacred ibis (Threskiornis aethiopicus), rosy-billed pochard (Netta peposaca)
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and black-bellied whistling duck (Dendrocygna autummalis) (Table 1).
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Salmonella spp. was not isolated from any of the 152 (0.0%; 95% CI: 0.0-1.9)
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pigeons analyzed. In contrast, in three faecal samples from the 44 captive zoo 8 Page 8 of 21
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animals tested (6.8%) positive isolation results were obtained (Table 1).
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In particular, Salmonella Panama was detected in wild boar (Sus scrofa), while
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Salmonella Apapa was isolated in milk snake (Lampropeltis triangulum). The
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Salmonella serovar isolated in ostrich could not be identified. Antibodies against T. gondii were observed in 9.2% of 142 (CI95%: 4.8-13.6)
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feral pigeons. Titers of 1:25, 1:50, 1:100 and 1:500 were detected in three (23.1%), six
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(46.2%), three (23.1%) and one (7.7%) pigeons, of the 13 seropositive animals,
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respectively. Statistically significant differences between the seropositivity against T.
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gondii and sex or age were not found. The overall seropositivity to T. gondii in zoo
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animals was 26.9% (29/108; CI95%: 19.6-34.1). Antibodies were detected in 42.9%
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carnivore (12/28), 30.2% ungulate (13/43) and 11.4% bird (4/35) samples, but were not
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detected in two primates analyzed. Titers of 1:25 were found in nine (31.0%), 1:50 in
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three (10.3%), 1:100 in 10 (34.5%) and 1:500 in seven (24.1%) of the 29 seropositive
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animals. Statistically significant differences between seropositivity and sex, age or
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group of species (birds, carnivores, ungulates or primates) were not observed.
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4. Discussion
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The results from the present study confirm local circulation of flavivirus,
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including WNV and USUV, in non-migratory feral pigeons in Córdoba city. Five out of
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11 seropositive pigeons detected by bELISA were negative in USUV, WNV and MBV
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VNT. The results could be due to the higher sensitivity of the bELISA test, to the
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detection of different types of antibodies using bELISA and VNTs, or to the circulation
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of other antigenically related flaviviruses [18]. Accordingly, circulation and mortality
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associated to Bagaza virus infections have been detected in wild bird species, including
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the genus Columba, in Spain [19,20]. Similar rates of seropositivity against WNV and 9 Page 9 of 21
USUV were detected in pigeons and zoo birds, which indicates a homogeneous
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circulation of these flaviviruses in these species. To date, the WNV outbreaks reported
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in horses (n= 88) and in humans (n= 2) in Spain have only been detected in the southern
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region [21]. Even though pigeons are included in the Spanish WNV Monitoring
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Program [22], to our knowledge, these are the first results in this species in Spain.
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Similar seroprevalences against flaviviruses were previously reported in both horses and
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wild birds in southern Spain [23,24,25], which suggests an endemic circulation of
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flaviviruses in this area. Higher seroprevalence (12.8%) to WNV was reported in game
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birds (partridges and pheasants) sampled in the same region where the highest number
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of outbreaks in horses in Spain was reported [26]. The seroprevalence to WNV in
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pigeons from other European countries ranges between 0 and 54% [27,14], and pigeons
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have been proposed as interesting sentinels of WNV transmission risk to humans [14].
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During the last few years, the number of USUV outbreaks reported in birds and humans
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has significantly increased in different European countries, including Spain [28,29,30].
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Seroprevalence obtained in pigeons in this study was in accordance with those
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previously reported in columbids and other wild bird species in Europe [26,14].
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Specific antibodies against WNV and USUV were confirmed in captive white
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stork and ostrich, respectively. Furthermore, one emperor goose showed specific
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neutralizing antibodies against the three flaviviruses tested using VNT. Although the
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higher titers against WNV detected in this species suggest infection with this flavivirus,
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contact with USUV and/or MBV cannot be ruled out. Interestingly, MBV circulation
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has been detected in gull populations in Spain [15]. Susceptibility to WNV infection in
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white stork and emperor goose has been previously confirmed [31,32]. Seroprevalence
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to USUV detected in zoo birds in our study (3.6%) was in accordance with that found in
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other zoos in Europe [33]. To the best of our knowledge, this is the first report of USUV
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infection in ostriches. None of the pigeons tested showed antibodies against AIV. Although
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susceptibility to AIV infection in columbids has been previously demonstrated [34], our
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results are consistent with that reported in columbid species in Spain [35] and other
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European countries [36,37]. Experimental studies have showed that pigeons could be
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considered as AIV “dead end” host because the shedding levels are below the threshold
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of the minimal infective particles required to infect other species. Therefore,
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disseminating AIV in this species is more likely to occur via mechanical route [34].
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AIV seropositivity was detected in five captive bird species, four of them belonging to
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the order Anseriformes, which are the main reservoirs of AIV [38]. To author’s
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knowledge, this is the first report of antibodies against AIV in African sacred ibis.
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The absence of Salmonella spp. isolation in pigeons suggests a limited role of
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feral pigeons in the transmission of this enteropathogen in the study area. These results
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are consistent with those previously reported in European countries including Spain
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[10,39,40]. Salmonella spp. was isolated in 3 out of 44 samples from zoo animals, in
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particular, in wild boar, milk snake and in ostrich. Due the intermittent shedding of
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Salmonella spp. in feces; the prevalence found might have been underestimated. The
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number of reports about Salmonella infections in humans associated to contact with
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reptiles has increased in the last years due to the use of these species as exotic pets [41].
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Also, clinical infections have been confirmed in zoo animals and zoo visitors by direct
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or indirect contact with reptiles infected with Salmonella spp. [42,43]. The serotype
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(Salmonella Apapa) detected in milk snake in the present study has been previously
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associated with sepsis in human neonates due to direct or indirect contact with bearded
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dragons [44]. Salmonella Panama was confirmed in one wild boar without clinical
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symptoms. Although this species is considered natural reservoir of Salmonella spp. in
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Spain [45] and this serotype has previously been isolated in domestic pig [46], this is
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the first report of Salmonella Panama infection in wild boar. Pigeons are a good model and indirectly indicate soil contamination by T. gondii
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oocysts, since they feed from the ground and they are highly susceptible to oral
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infection with oocysts [47]. Importantly, an association of the consumption of pigeon
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meat with T. gondii infection has been reported [48]. There have been many reports of
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T. gondii antibodies in feral pigeons, and the parasite has been isolated from this species
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[48] but to our knowledge there have not been previous reports of seroprevalence of T.
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gondii in feral pigeons in Spain. Seroprevalence levels observed in the present study
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(less than 10%) are in agreement with most of the previously reported studies in pigeons
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worldwide [49]. The wide geographic distribution of Columba livia, the ecological
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association to human settlements and the sedentary behavior suggest the pigeon as good
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sentinel species for the presence of oocysts in the human environment. Additional
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studies are required in order to investigate the role of pigeons in the epidemiology of
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T. gondii.
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Widespread T. gondii seroprevalence was observed in captive zoological species
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in CMZP. Sixteen of 33 (48.5%) zoological species analyzed had at least one
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seropositive animal to T. gondii with 26.9% as average seroprevalence. This study is the
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first report of presence of T. gondii antibodies in European mink, a critically endangered
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species [50]. High seroprevalence was observed in the felid species (46.2%), and
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importantly all the samples analyzed from lions, jaguars and Bengal tigers were
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positive. These results could be of Animal Health and Conservation concern in CMZP
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due to the fact that seropositive felids might shed oocysts and the high susceptibility to
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T. gondii infection of some zoo species such as Marsupials and New World monkeys
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[49,51,52]. In the present study, antibodies against T. gondii were widespread in captive-
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ungulates, being observed in 6 of 9 species analyzed. In Spain, presence of such
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antibodies has been previously described in free-living wild ruminants [53] and in wild
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boars [54]. Birds showed lower seroprevalence levels and antibodies were detected in
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only 4 of 14 species analyzed. The prevalence of antibodies against T. gondii in wild
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bird species in Spain has been highly heterogeneous, depending on their feeding
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behaviour and geographical location with higher levels in carnivore wild birds [55].
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Low number of carnivore birds was included in the present study, which could be one
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reason for the low prevalence observed. Another explanation could be low exposure
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to oocysts in the environment. In addition, some recent studies have suggested that
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lower sensitivity of serological tests, such as MAT, in birds may sub-estimate
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seropositivity levels in these species [56].
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The results obtained in the present study suggest a limited role of pigeons in the
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transmission of AIVs and Salmonella spp. However, the seroprevalence against
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flaviviruses and T. gondii found in this species, as well the presence of flaviviruses,
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AIV, Salmonella spp. and T. gondii detected in captive zoo animals, indicate the need
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to monitor these species in the study area. Control measures, including monitoring,
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population control, vaccination programs and management measures should be
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maintained and/or implemented in order to prevent transmission of these zoonotic
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agents between pigeons and sympatric species, included human.
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Acknowledgements 13 Page 13 of 21
We would like to thank the Municipal Council of Córdoba for providing
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technical support. We thank Carlos Expósito in the samples collection, Cécile Beck and
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Steeve Lowenski in the virus neutralization tests, Maria Puig in the modified
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agglutination tests, and Carmen Borge and Estefanía Jurado in the Salmonella
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isolations.
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References
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[1] R. Sacchi, A. Gentilli, E. Razzetti, F. Barbieri, Effects of building features on density and flock distribution of feral pigeons Columba livia var. domestica in an urban environment, Can. J. Zool. 80 (2002) 48-54.
M
[2] J.C. Senar, J. Carrillo, L. Arroyo, T. Montalvo, V. Peracho, Estima de la abundancia de palomas (Columba livia var.) de la ciudad de Barcelona y valoración de la efectividad del control por eliminación de individuos, Arx. misc. zool. 7 (2009) 62–71.
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[3] I. Amoruso, L. Fabbris, M. Mazza, G. Caravello, Estimation of Feral Pigeon (Columba livia) population size using a novel Superimposed Urban Strata (SUS) method, Urban Ecosyst. 17 (2014) 597-612. [4] P. Albonetti, A. Marletta, I. Repetto, E.A. Sasso, Efficacy of nicarbazin (Ovistop®) in the containment and reduction of the populations of feral pigeons (Columba livia var. domestica) in the city of Genoa, Italy: a retrospective evaluation, Vet Ital. 51 (2015) 6372.
Ac ce pt e
307 308 309 310 311 312 313 314 315 316 317 318 319 320 321 322 323 324 325 326 327 328 329 330 331 332 333 334 335 336 337 338 339 340 341
cr
306
[5] T. Mcnamara, The role of zoos in biosurveillance, Int. Zoo Yb. 41 (2007) 12-15. [6] A. Dovc, O. Zorman-Rojs, A. Vergles-Rataj, V. Bole-Hribovsek, U. Krapez, M. Dobeic, Health status of free-living pigeons (Columba livia domestica) in the city of Ljubljana, Acta Vet Hung. 52 (2004) 219-226. [7] E. De-Sousa, A.J. Berchieri, A.A. Pinto, R.Z. Machado, A.O. de Carrasco, J.A. Marciano, K. Werther, Prevalence of Salmonella spp. Antibodies to Toxoplasma gondii, and Newcatle disease virus in feral pigeons (Columba livia) in the city of Jaboticabal, Brazil, J Zoo Wildl Med. 41 (2010) 603-607. [8] J. Salinas, M.R. Caro, F. Cuello, Antibody prevalence and isolation of Chlamydia psittaci from pigeons (Columba livia), Avian Dis. 37 (1993) 523-7. [9] B. Vázquez, F. Esperon, E. Neves, J. Lopez, C. Ballesteros, M.J. Munoz, Screening for several potential pathogens in feral pigeons (Columba livia) in Madrid, Acta Vet Scand. 52 (2010) 45. 14 Page 14 of 21
[10] L. Casanovas, M. de Simón, M.D. Ferrer, J. Arqués, G. Monzón, Intestinal carriage of campylobacters, salmonellas, yersinias and listerias in pigeons in the city of Barcelona, J Appl Bacteriol. 78 (1995) 11-3. [11] M.F. Colom-Valiente, M. Alberdi, I. Meseguer, J.M. Torres-Rodríguez, Isolation of Cryptococcus neoformans from environmental samples in Alicante, Rev Iberoam Micol. 14 (1997) 63-4.
ip t
[12] B. Close, K. Banister, V. Baumans, E.M. Bernoth, N. Bromage, J. Bunyan, W. Erhardt, P. Flecknell, N. Gregory, H. Hackbarth, D. Morton, C. Warwick, Recommendations for euthanasia of experimental animals: Part 2. Lab. Lab Anim. 31 (1997) 1-32.
us
cr
[13] D. Gibbs, E. Barnes, C. John, Pigeons and Doves: A Guide to the Pigeons and Doves of the World, Christppher Helm Publishers, London, 2010.
an
[14] S.C. Chaintoutis, C.I. Dovas, M. Papanastassopoulou, S. Gewehr, K. Danis, C. Beck, S. Lecollinet, V. Antalis, S. Kalaitzopoulou, T. Panagiotopoulos, S. Mourelatos, S. Zientara, O. Papadopoulos, Evaluation of a West Nile virus surveillance and early warning system in Greece, based on domestic pigeons, Comp Immunol Microbiol Infect Dis. 37 (2014) 131-41.
d
M
[15] A. Arnal, E. Gómez-Díaz, M. Cerdà-Cuéllar, S. Lecollinet, J. Pearce-Duvet, N. Busquets, I. García-Bocanegra, N. Pagès, M. Vittecoq, A. Hammouda, B. Samraoui, R. Garnier, R. Ramos, S. Selmi, J. González-Solís, E. Jourdain, T. Boulinier, Circulation of a Meaban-like virus in yellow-legged gulls and seabird ticks in the western Mediterranean basin, PLoS One 13 (2014) e89601. [16] M.Y. Popoff, J. Bockemühl, F.W. Brenner, L.L. Gheesling, Supplement 2000 (no. 44) to the Kauffmann-White scheme, Res Microbiol. 152 (2001) 907-9.
Ac ce pt e
342 343 344 345 346 347 348 349 350 351 352 353 354 355 356 357 358 359 360 361 362 363 364 365 366 367 368 369 370 371 372 373 374 375 376 377 378 379 380 381 382 383 384 385 386 387 388 389 390 391
[17] J.P. Dubey, G. Desmonts, Serological responses of equids fed Toxoplasma gondii oocysts, Equine.Vet. J. 19(4) (1987) 337–339. [18] B.J. Blitvich, N.L. Marlenee, R.A. Hall, C.H. Calisher, R.A. Bowen, J.T. Roehring, N. Komar, S.A. Langevin, B.J. Beaty, Epitope-blocking enzyme-linked immunosorbent assays for the detection of serum antibodies to West Nile virus in multiple avian species, J Clin Microbiol. 41 (2003) 1041-7. [19] V. Gamino, A.V. Gutiérrez-Guzmán, I.G. Fernández-de-Mera, J.A. Ortíz, M. Durán-Martín, J. De la Fuente, C. Gortázar, U. Höfle, Natural Bagaza virus infection in game birds in southern Spain, Vet Res. 43 (2012) 65. [20] I. García-Bocanegra, I. Zorrilla, E. Rodríguez, E. Rayas, L. Camacho, I. Redondo, F. Gómez-Guillamón, Monitoring of the Bagaza virus epidemic in wild birds species in Spain, 2010, Transbound Emerg Dis. 60 (2013) 120-6. [21] RASVE (Red de Alerta Sanitaria Veterinaria), 2015. Available at: http://rasve.magrama.es/RASVE_2008/Publica/Focos/Consulta.aspx (accessed April 15, 2015). 15 Page 15 of 21
[22] RASVE (Red de Alerta Sanitaria Veterinaria), 2015. Available at: http://rasve.magrama.es/Publica/Programas/Normativa.asp (accessed April 25, 2015). [23] I. García-Bocanegra, N. Busquets, S. Napp, A. Alba, I. Zorrilla, R. Villalba, A. Arenas, Serosurvey of West Nile Virus and Other Flaviviruses of the Japanese Encephalitis Antigenic Complex in Birds from Andalusia, Southern Spain, Vector Borne Zoonotic Dis. 11 (2011) 1107-1113.
ip t
[24] I. García-Bocanegra, J.A. Jaén-Téllez, S. Napp, A. Arenas-Montes, M. FernándezMorente, V. Fernández-Molera, A. Arenas, Monitoring of the West Nile virus epidemic in Spain between 2010 and 2011, Transbound Emerg Dis. 59 (2012) 448-55.
us
cr
[25] G. López, M.A. Jiménez-Clavero, A. Vázquez, R. Soriguer, C. Gómez-Tejedor, A. Tenorio, J. Figuerola, Incidence of West Nile virus in birds arriving in wildlife rehabilitation centers in southern Spain, Vector Borne Zoonotic Dis. 11 (2011) 285-90.
an
[26] F. Llorente, E. Pérez-Ramírez, J. Fernández-Pinero, R. Soriguer, J. Figuerola, M.A. Jiménez-Clavero, Flaviviruses in game birds, southern Spain, 2011-2012, Emerg Infect Dis. 19 (2013) 023-5.
M
[27] P. Lena, E. Bureau, S. Reynard, P. Sabatier, S. Bruyas, H. Zeller, Sero-survey for West Nile in wild birds in "The Dombes" Wetlands (01, Ain. France) in 2001: absence of detection of viral activity, Rev Med Vet-Toulouse. 157 (2006) 612-618.
d
[28] H. Weissenböck, J. Kolodziejek, A. Url, H. Lussy, B. Rebel-Bauder, N. Nowotnyz, Emergence of Usutu virus, an African mosquito-borne flavivirus of the Japanese encephalitis virus group, central Europe, Emerg Infect Dis. 8 (2002) 652-6.
Ac ce pt e
392 393 394 395 396 397 398 399 400 401 402 403 404 405 406 407 408 409 410 411 412 413 414 415 416 417 418 419 420 421 422 423 424 425 426 427 428 429 430 431 432 433 434 435 436 437 438 439 440 441
[29] U. Höfle, V. Gamino, I.G. Fernández de Mera, A.J. Mangold, J.A. Ortíz, J. de la Fuente, Usutu Virus in Migratory Song Thrushes, Spain, Emerg Infect Dis. 19 (2013) 1023–1025. [30] M. Santini, T. Vilibic-Cavlek, B. Barsic, L. Barbic, V. Savic, V. Stevanovic, E. Listes, A. Di Gennaro, G. Savini, First cases of human Usutu virus neuroinvasive infection in Croatia, August-September 2013: clinical and laboratory features, J Neurovirol. 21 (2015) 92-7. [31] M. Malkinson, Y. Weisman, S. Pokamonski, R. King, V. Deubel, Intercontinental transmission of West Nile virus by migrating white storks, Emerg Infect Dis. 7 (2001) 540. [32] CDC (Centers for Diseases Control and Prevention), 2015. Available at: http://www.cdc.gov/westnile/resources/pdfs/Bird%20Species%201999-2012.pdf (accessed May 10, 2015). [33] N. Buchebner, W. Zenker, C. Wenker, H.W. Steinmetz, E. Sós, N. Nowotny, Low Usutu virus seroprevalence in four zoological gardens in central Europe, BMC Vet Res. 9 (2013) 153. [34] C. Abolnik, A current review of avian influenza in pigeons and doves (Columbidae), Vet Microbiol. 170 (2014) 181–196. 16 Page 16 of 21
[35] E. Pérez-Ramírez, X. Gerrikagoitia, M. Barral, U. Höfle, Detection of low pathogenic avian influenza viruses in wild birds in Castilla-La Mancha (south central Spain), Vet Microbiol. 146 (2010) 200-208.
ip t
[36] A. Lillehaug, C. Monceyron-Jonassen, B. Bergsjø, M. Hofshagen, J. Tharaldsen, L.L. Nesse, K. Handeland, Screening of feral pigeon (Columba livia), mallard (Anas platyrhynchos) and graylag goose (Anser anser) populations for Campylobacter spp., Salmonella spp., avian influenza virus and avian paramyxovirus, Acta Vet Scand. 46 (2005) 193-202.
cr
[37] L. Teske, M. Ryll, S. Rautenschlein, Epidemiological investigations on the role of clinically healthy racing pigeons as a reservoir for avian paramyxovirus-1 and avian influenza virus, Avian Pathol. 42 (2013) 557-65.
us
[38] J.K. Kim, N.J. Negovetich, H.L. Forrest, R.G. Webster, Ducks: the “Trojan horses” of H5N1 influenza, Influenza Other Resp. 3 (2009) 121-8.
an
[39] L. Teske, M. Ryll, D. Rubbenstroth, I. Hänel, M. Hartmann, L. Krejenbrock, S. Rautenschlein. Epidemiological investigations on the possible risk of distribution of zoonotic bacteria through apparently healthy homing pigeons, Avian Pathol. 42 (2013) 397-407.
d
M
[40] A. Gargiulo, T.P. Russo, R. Schettini, K. Mallardo, M. Calabria, L.F. Menna, P. Raia, U. Pagnini, V. Caputo, A. Fioretti, L. Dipineto, Occurrence of enteropathogenic bacteria in urban pigeons (Columba livia) in Italy, Vector Borne Zoonotic Dis. 14 (2014) 251-5. [41] L. Geue, U. Löschner, Salmonella enterica in reptiles of German and Austrian origin, Vet Microbiol. 84 (2002) 79–91.
Ac ce pt e
442 443 444 445 446 447 448 449 450 451 452 453 454 455 456 457 458 459 460 461 462 463 464 465 466 467 468 469 470 471 472 473 474 475 476 477 478 479 480 481 482 483 484 485 486 487 488 489 490 491
[42] C.R. Friedman, C. Torigian, P.J. Shillam, R.E. Hoffman, D. Heltzel, J.L. Beebe, G. Malcolm, W.E. Dewitt, L. Hutwagner, P.M. Griffin, An outbreak of salmonellosis among children attending a reptile exhibit at a zoo, J Pediatr. 132 (1998) 802–807. [43] L. Bauwens, F. Vercammen, S. Bertrand, J.M. Collard, S. De Ceuster, Isolation of Salmonella from environmental samples collected in the reptile department of Antwerp Zoo using different selective methods, J Appl Microbiol. 101 (2006) 284–289. [44] R. Haase, T. Beier, M. Bernstadt, N. Merkel, J. Bartnicki, Neonatal infection with Salmonella apapa after contact with a reptile in the home, Z. Geburtsh Neonatol. 215 (2011) 86-88. [45] N. Navarro-González, G. Mentaberre, C.M. Porrero, E. Serrano, A. Mateos, J.M. López-Martín, S. Lavín, L. Domínguez, Effect of Cattle on Salmonella Carriage, Diversity and Antimicrobial Resistance in Free-Ranging Wild Boar (Sus scrofa) in Northeastern Spain, PLoS One 7 (2012) e51614. [46] J.D. Kich, A. Coldebella, N. Mores, M. Nogueira, M. Cardoso, P.M. Fratamico, J.E. Call, P. Fedorka-Cray, J.B. Luchansky, Prevalence, distribution, and molecular 17 Page 17 of 21
characterization of Salmonella recovered from swine finishing herds and a slaughter facility in Santa Catarina, Brazil, Int J Food Microbiol. 151 (2011) 307-13. [47] H. Waap, R. Cardoso, A. Leitão, T. Nunes, A. Vilares, M.J. Gargaté, J. Meireles, H. Cortes, H. Ângelo, In vitro isolation and seroprevalence of Toxoplasma gondii in stray cats and pigeons in Lisbon, Portugal, Vet Parasitol. 187 (2012) 542-7.
cr
ip t
[48] C. Alvarado-Esquivel, O. Liesenfeld, B.D. Burciaga-López, A. Ramos-Nevárez, S. Estrada-Martínez, S.M. Cerrillo-Soto, F.A. Carrete-Ramírez, Mde.L. López-Centeno, M.M. Ruiz-Martínez, Seroepidemiology of Toxoplasma gondii infection in elderly people in a northern Mexican city, Vector Borne Zoonotic Dis. 12 (2012) 568-74. [49] J.P. Dubey, Toxoplasmosis in animals and humans, 2nd ed., CRC Press, Boca Raton, FL, 2010.
us
[50] UICN (International Union for Conservation of Nature), 2015. Available at: http://www.iucnredlist.org/details/14018/0 (accessed April 5, 2015).
an
[51] C. Juan-Sallés, N. Prats, A.J. Marco, J.A. Ramos-Vara, D. Borrás, J. Fernández, Fatal acute toxoplasmosis in three golden lion tamarins (Leontopithecus rosalia), J Zoo Wildl Med. 29 (1998) 55-60.
M
[52] X. Fernández-Aguilar, D. Ajzenberg, O. Cabezón, A. Martínez-López, L. Darwich, J.P. Dubey, S. Almería, Fatal toxoplasmosis associated with an atypical Toxoplasma gondii strain in a Bennett's wallaby (Macropus rufogriseus) in Spain, Vet Parasitol. 196 (2013) 523-527.
d
[53] C.B. Gauss, J.P. Dubey, D. Vidal, O. Cabezón, F. Ruiz-Fons, J. Vicente, I. Marco, S. Lavin, C. Gortazar, S. Almería, Prevalence of Toxoplasma gondii antibodies in red deer (Cervus elaphus) and other wild ruminants from Spain, Vet Parasitol. 136 (2006) 193-200.
Ac ce pt e
492 493 494 495 496 497 498 499 500 501 502 503 504 505 506 507 508 509 510 511 512 513 514 515 516 517 518 519 520 521 522 523 524 525 526 527 528 529 530 531 532 533 534 535 536 537 538
[54] C.B. Gauss, J.P. Dubey, D. Vidal, F. Ruiz, J. Vicente, I. Marco, S. Lavin, C. Gortazar, S. Almería, Seroprevalence of Toxoplasma gondii in wild pigs (Sus scrofa) from Spain, Vet Parasitol. 131 (2005) 151-6. [55] O. Cabezón, I. García-Bocanegra, R. Molina-López, I. Marco, J.M. Blanco, U. Höfle, A. Margalida, E. Bach-Raich, L. Darwich, I. Echeverría, E. Obón, M. Hernández, S. Lavín, J.P. Dubey, S. Almería, Seropositivity and risk factors associated with Toxoplasma gondii infection in wild birds from Spain, PLoS One 6 (2011) e29549. [56] L. Casartelli-Alves, V.C. Boechat, R. Macedo-Couto, L.C. Ferreira, J.L. Nicolau, L.B. Neves, P.R. Millar, R.T. Vicente, R.V. Oliveira, A.G. Muniz, I.C. Bonna, M.R. Amendoeira, R.C. Silva, H. Langoni, T.M. Schubach, R.C. Menezes, Sensitivity and specificity of serological tests, histopathology and immunohistochemistry for detection of Toxoplasma gondii infection in domestic chickens, Vet Parasitol. 204 (2014) 34651.
18 Page 18 of 21
Tables
539
Table 1. Prevalence of selected zoonotic pathogens in feral pigeons (Columba livia var.
540
domestica) and sympatric zoological species in Córdoba, Southern Spain.
Ac ce pt e
d
M
an
us
cr
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538
19 Page 19 of 21
Salmonella spp.
Toxoplasma gondii
0/148
0/152
13/142 (9.2%)
0/3 0/7 0/1 0/5 0/3 1/1 (100 %) 1/20 (5%)
-
0/1 0/1 0/1 0/1 1/1 (100 %) 0/1 1/6 (16.7%)
2/10 (20%) 3/15 (20%) 1/1(100%) 4/6 (66.7%) 2/3 (66.7%) 0/1 1/3 (33.3%) 0/3 13/42 (31%)
0/1 0/1
-
0/3 0/1 0/4
0/1 0/1
0/1 0/3 0/4 0/3 1/2 (50%) 1/13 (7.7%)
1/1 (100 %) 1/3 (33.3 %) 1/4 (25 %) 0/3 1/2 (50 %) 4/13 (30.8%)
0/1 0/1 0/2
0/1 0/3 1/4 (25 %) 1/11 (9.1%) 1/2 (50%) 3/21 (14.3%)
0/1 0/1 0/2
0/1 0/1 0/1 0/1 0/4
0/1 1/1 (100%) 1/2 (50%)
0/1
0/1
0/1
0/1
0/2 0/4 1/2 (50 %) 1/8 (12.5%)
0/2 0/4 0/2 0/8
0/1 0/1 1/2 1/4 (25%)
0/2 0/4 0/2 0/8
1/2 (50 %)
0/2
-
0/1
0/2
1/2 (50 %)
-
0/1
-
-
-
0/1
-
-
0/1 0/2 0/1 0/1 0/5
2/2 (100%) 1/1 (100%) 3/3 (100%) 0/7 3/3 (100%) 0/1 1/8 (12.5%) 2/3 (66.6%) 12/28 (42.9%)
-
-
0/2 0/1 0/1 0/1 0/2 0/2 0/2 0/1 0/1 0/1 0/14
0/1 0/1 0/2
-
-
0/1 0/1 0/2
-
-
-
0/1 1/1 (100 %) 1/2 (50 %)
4/49 (8.2%)
5/28 (17.9 %)
0/2 3/44 (6.8%)
M
us
-
an
11/142 (7.8 %)
ip t
Influenza
cr
Flaviviruses
Ac ce pt e
d
SPECIES ORDER COLUMBIFORMES Feral pigeon (Columba livia var. domestica) ORDER ARTIODACTYLA Aoudad (Ammotragus lervia) Red deer (Cervus elaphus) Fallow deer (Dama dama) Mouflon (Ovis musimon) Collared Peccary (Pecari tajacu) Black Wildebeest (Connochaetes gnou) Roe deer (Capreolus capreolus) Wild boar (Sus scrofa) Llama (Lama glama) Sheep (Ovis aries) Goat (Capra hircus) TOTAL ORDER PERISSODACTYLA Plains zebra (Equus burchellii) Donkey (Equus africanus asinus) Lowland Tapir (Tapirus terrestris) TOTAL ORDER ANSERIFORMES Rosy-billed Pochard (Netta peposaca) Black-bellied whistling duck (Dendrocygna autummalis) Mallard (Anas platyrhynchos) Bar-headed goose (Anser indicus) Emperor goose (Chen canagica) TOTAL ORDER ACCIPITRIFORMES Griffon vulture (Gyps fulvus) Booted eagle (Hieraaetus pennatus) Short-toed snake eagle (Circaetus gallicus) Common buzzard (Buteo buteo) TOTAL ORDER GALLIFORMES Indian peafowl (Pavo cristatus) ORDER STRUTHIONIFORMES Greater rhea (Rhea americana) Emu (Dromaius novaehollandiae) Ostrich (Struthio camelus) TOTAL ORDER CICONIIFORMES White stork (Ciconia ciconia) ORDER PELECANIFORMES African sacred ibis (Threskiornis aethiopicus) ORDER PSITTACIFORMES Muir's corella (Cacatua pastinator) ORDER CARNIVORA Lion (Panthera leo) Jaguar (Panthera onca) Bengal tiger (Panthera tigris tigris) Eurasian lynx (Lynx lynx) Brown bear (Ursus arctos pyrenaicus) European otter (Lutra lutra) Iberian wolf (Canis lupus signatus) European mink (Mustela lutreola) TOTAL ORDER PRIMATES Northern white-cheeked gibbon (Hylobates leucogenys) Brown lemur (Eulemur fulvus) Black-and-white ruffed lemur (Varecia variegata) Ring-tailed lemur (Lemur catta) Barbary macaque (Macaca sylvanus) White-collared Mangabey (Cercocebus atys ssp. lunulatus) De Brazza’s Monkey (Cercopithecus neglectus) White-tufted-ear Marmoset (Callithrix jacchus) Western Pygmy Marmoset (Callithrix pygmaea) South American Squirrel Monkey (Saimiri sciureus) TOTAL ORDER RODENTIA Patagonian Cavy (Dolichotis patagonum) Capybara (Hydrochoerus hydrochaeris) TOTAL ORDER SQUAMATA Corn snake (Elaphe guttata) Milk snake (Lampropeltis triangulum) TOTAL ORDER TESTUDINES African spurred tortoise (Geochelone sulcata) TOTAL IN CAPTIVE ANIMALS
-20 -
Page 20 of 21
29/108 (26.9%)
541 542
Table 2. Results of virus neutralization test for the detection of flaviviruses in feral
543
pigeons and captive zoo animals. Species, (no.)
VNT titers WNV
USUV
MBV
Feral pigeon (n= 1)
<10
10
<20
Feral pigeon (n= 1)
20
<10
<20
Feral pigeon (n= 1)
80
<10
<20
WNV
Feral pigeon (n= 1)
<10
10
<20
USUV
Feral pigeon (n= 1)
80
10
<20
WNV
Feral pigeon (n= 1)
<10
20-40
<20
USUV
Feral pigeon (n= 5)
<10
Total (%)
3(2.1)
ip t
Feral pigeon (Columba livia) (n=142)
USUV
an
us
cr
WNV
<20
3(2.1)
0
Undetermined Flavivirus
M
<10
Emperor goose (Chen canagica) (n= 1)
80
40
20
Undetermined Flavivirus
White stork (Ciconia ciconia) (n= 1)
40
<10
NA1
WNV
<10
20
<20
USUV
<10
<10
<20
Undetermined Flavivirus
1(2.0)
1(2.0)
Ac ce pt e
Ostrich (Rhea americana) (n= 1)
d
Zoo animals (n=49)
Black wildebeest (Connochaetes gnou) (n= 1) Total (%)
544 545
Interpretation
WNV, West Nile virus; USUV, Usutu virus; MV, Meaban virus; VNT, virus neutralization test; 1. NA: Not analyzed, low volume.
21 Page 21 of 21
S0147-9571(15)00078-8 http://dx.doi.org/doi:10.1016/j.cimid.2015.10.003 CIMID 1030
To appear in: Received date: Revised date: Accepted date:
22-6-2015 23-9-2015 17-10-2015
Please cite this article as: Cano-Terriza D, Guerra R, Lecollinet S, Cerdgravea-Cu´ellar M, Cabez´on O, Almer´ia S, Garc´ia-Bocanegra I, Epidemiological survey of zoonotic pathogens in feral pigeons (Columba livia var. domestica) and sympatric zoo species in Southern Spain., Comparative Immunology, Microbiology and Infectious Diseases (2015), http://dx.doi.org/10.1016/j.cimid.2015.10.003 This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and review of the resulting proof before it is published in its final form. Please note that during the production process errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain.
1
Epidemiological survey of zoonotic pathogens in feral pigeons (Columba livia var.
2
domestica) and sympatric zoo species in Southern Spain.
3 Authors
5
David Cano-Terrizaa, Rafael Guerrab, Silvie Lecollinetc, Marta Cerdà-Cuéllard, Oscar
6
Cabezóne, Sonia Almeríad,f, Ignacio García-Bocanegraa *
ip t
4
cr
7 8
a
9
Excellence International Campus (ceiA3), Campus de Rabanales, 14071, Córdoba,
us
Dept. de Sanidad Animal, Facultad de Veterinaria, Universidad de Córdoba-Agrifood
Spain.
11
b
12
Córdoba, Spain.
13
c
14
INRA, ANSES, ENVA, Maisons-Alfort, F-94703, France.
15
d
16
Agroalimentàries (IRTA), Campus de la Universitat Autònoma de Barcelona,
17
Bellaterra, Barcelona, Spain.
18
e
19
Animals, Universitat Autònoma de Barcelona, 08193, Bellaterra, Barcelona, Spain.
20
f
21
de Barcelona, 08193, Bellaterra, Barcelona, Spain.
an
10
M
Parque Zoológico Municipal de Córdoba (PZMC), Avenida Linneo s/n, 14071,
d
ANSES, Laboratoire de Santé Animale de Maisons-Alfort, UMR 1161 Virologie,
Ac ce pt e
Centre de Recerca en Sanitat Animal (CReSA) - Institut de Recerca i Tecnologia
Servei d'Ecopatologia de Fauna Salvatge (SEFaS), Dept. de Medicina i Cirugia
Dept. de Sanitat i d’Anatomia Animals, Facultat de Veterinària, Universitat Autònoma
22 23
* Corresponding author at: Departamento de Sanidad Animal (Universidad de
24
Córdoba). Edificio Sanidad Animal 2ª planta, Campus Universitario Rabanales, 14071,
25
Córdoba, Spain. Tel: +34 95 721 8725; fax: +34 95 721 8727.
26
E-mail address: [email protected] (Ignacio García-Bocanegra) 1 Page 1 of 21
27
Abstract A cross-sectional study was carried out to determine the prevalence of
29
pathogenic zoonotic agents (flaviviruses, avian influenza viruses (AIVs), Salmonella
30
spp. and Toxoplasma gondii) in feral pigeons and sympatric zoo animals from Córdoba
31
(Southern Spain) between 2013 and 2014. Antibodies against flaviviruses were
32
detected in 7.8% out of 142 (CI95%: 3.7-11.8) pigeons, and 8.2% of 49 (CI95%: 0.9-
33
15.4) of zoo animals tested. Antibodies with specificity against West Nile virus
34
(WNV) and Usutu virus (USUV) were confirmed both in pigeons and in zoo birds.
35
Even though seropositivity to AIVs was not detected in any of the analyzed pigeons,
36
17.9% of 28 (CI95%: 3.7-32.0) zoo birds tested showed positive results. Salmonella spp.
37
was not isolated in any of 152 faecal samples collected from pigeons, while 6.8% of 44
38
zoo animals were positive. Antibodies against T. gondii were found in 9.2% of 142
39
(CI95%: 4.8-13.6) feral pigeons and 26.9% of 108 (CI95%: 19.6-34.1) zoo animals. This
40
is the first study on flaviviruses and T. gondii in feral pigeons and captive zoo species
41
in Spain. Antibodies against WNV and USUV detected in non-migratory pigeons and
42
captive zoo animals indicate local circulation of these emerging pathogens in the study
43
area. T. gondii was widespread in species analyzed. This finding could be of
44
importance for Public Health and Conservation of endangered species present in zoo
45
parks. Pigeons and zoo animals may be included as sentinel species for monitoring
46
zoonotic pathogens in urban areas.
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Keywords: Avian influenza viruses; Captive zoo animals; Feral pigeons; Flaviviruses;
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Salmonella spp.; Toxoplasma gondii.
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Highlights
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Spain •
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•
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these species
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Circulation of Flaviviruses, AIV, Salmonella spp. and T. gondii were detected in
The results obtained could have relevance for Public Health and Conservation
Pigeons and zoo animals may be included as
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We report prevalence of zoonotic pathogens in feral pigeons and zoo animals in
sentinel species for monitoring zoonotic pathogens in urban areas
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1. Introduction Feral pigeon (Columba livia var. domestica; Columbidae family) is one of the
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bird species most frequently found in urban and peri-urban areas. During the last
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decades, their populations have increased exponentially in different countries, reaching
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densities higher than 2000 birds/km2 in many European cities [1,2]. Favorable
65
environmental conditions and abundant food, which imply loosing the breeding season,
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and the absence of predators are the main factors implicated in the high increase of their
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populations [3]. Feral pigeon is considered a pest species in many urban areas due to the
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destruction architectonic and urban heritage, damage to agriculture and risk of disease
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transmission to other sympatric species, including humans. In this sense, control
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measures including surgical or chemical sterilization, removal of eggs, physical
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repellents, use of birds of prey or capturing and controlled elimination are frequently
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implemented in urban areas [2,4].
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Zoological parks are a very favorable habitat for pigeons, where they are found
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in higher densities than in urban areas. High populations imply significant economic
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losses associated to the consumption of food intended for captive zoo animals.
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Furthermore, the risk of disease transmission from pigeons to zoo sympatric species can
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be of concern for Animal Health and Conservation. In this sense, the importance of zoos
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for surveillance of zoonotic and emerging diseases has been previously suggested [5].
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Even though feral pigeons are considered reservoirs of zoonotic diseases
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including
Newcastle
disease,
histoplasmosis,
ornithosis,
salmonellosis,
or
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cryptococcosis [6,7], information about zoonotic diseases in this species in Spain is yet
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very limited. High prevalence of Chlamydophila psittaci infection was found in central
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(Madrid: 52.6%) and southwest (Murcia: 35.9%) areas [8,9]. Prevalence of 4 Page 4 of 21
Campylobacter spp. infection ranged between 1.1% and 69.1% in Barcelona and
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Madrid, respectively [10,9]. Also, 1.5% of Salmonella spp. and 0.25% of Yersinia
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intermedia infections was detected in pigeons from Barcelona city [10]. In addition,
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70.3% of pigeons analyzed in Alicante (southwest Spain) showed Cryptococcus
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neoformans infection [11].
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The aim of this study was to determine the prevalence of selected zoonotic
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pathogens (flaviviruses, avian influenza viruses (AIVs), Salmonella spp. and T. gondii)
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in feral pigeons, and to assess the risk of transmission to sympatric zoological species in
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the Córdoba Municipal Zoo Park (CMZP, Southern Spain).
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2. Materials and Methods
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2.1 Study design and sampling
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A total of 152 feral pigeons were captured in the CMZP (Southern Spain)
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between November 2013 and May 2014. Captures were performed within the course of
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a pest control program undertaken by the local authorities following European
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guidelines (UNE 171210: 2008). Animals were handled and sampled according to
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regulations of the Regional Government of Andalusia. Blood samples were taken from the brachial vein, after those animals were
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humanely euthanized by overdose sodium pentobarbital (Dolethal®, Vetoquinol
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Laboratories, Lure, France) (0.5 ml/kg) as recommended for birds [12]. Samples
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were placed into sterile tubes without anticoagulant and centrifuged at 3000 rpm for 10
105
min. Sera were stored at -20ºC until analysis. All individuals were necropsied and
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inspected for the presence of macroscopic lesions. Pigeons were classified as juveniles
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(< 8 months) and adult (> 8 months) according to previously established criteria [13]. 5 Page 5 of 21
Sera from 108 zoo animals belonging to 34 species (Table 1), which share habitat with
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the sampled pigeons, were also collected. In addition, digestive content (2-3 gr) from all
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pigeons and 44 fresh faecal samples from 35 different zoo species were collected for
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Salmonella spp. isolation.
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2.2 Laboratory analysis
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Sera from 142 pigeons and 49 captive zoo animals were tested for antibodies
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against one epitope of the preMembrane-Envelope (prM-E) protein common to
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flaviviruses of the Japanese Encephalitis antigenic complex (JEV) using a commercial
116
blocking ELISA (bELISA 10.WNV.K3 INGEZIM West Nile COMPAC®, Ingenasa,
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Madrid, Spain). ELISA-positive sera were confirmed by virus neutralization test (VNT)
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for the detection of specific neutralizing antibodies against West Nile virus (WNV; Is98
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strain), Usutu virus (USUV; It12 strain) and Meaban virus (MBV; Brest ART707
120
strain). VNTs were performed as previously described [14,15]. Samples that showed
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neutralization and absence of cytopathic effect at dilutions ≥ 10 for WNV and USUV
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and ≥ 20 for MBV, were considered positive. Interpretation of results was based on
123
comparison of VNT titers obtained in parallel against the 3 flaviviruses. The
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neutralizing immune response observed was considered specific when VNT titers for a
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given virus was ≥ 4-fold higher than titers obtained for the other viruses. Samples
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showing VNT titers differences ≤ 2-fold between the viruses examined were considered
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positive for flavivirus but not conclusive for any specific virus.
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The presence of antibodies against the nucleoprotein of AIVs type A was
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determined using a commercial (bELISA) (1.0.FLU.K3 INGEZIM INFLUENZA A®,
130
Ingenasa, Madrid, Spain) according to the manufacturer´s recommendations. A total of
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148 pigeons and 28 zoo birds were analyzed. 6 Page 6 of 21
Faecal droppings and faecal samples were processed as described in Annex D of
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the Spanish standard UNE-EN ISO 6579:2002 for Salmonella isolation. Salmonella-
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presumptive colonies were tested with the Mucap (Biolife, Milano, Italy) and indole
135
tests. Salmonella serotyping was carried out according to the Kauffman-White scheme
136
[16] by the Agri-food laboratory of Departament d’Agricultura, Ramaderia, Pesca,
137
Alimentació i Medi Natural (Generalitat de Catalunya, Spain).
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Sera from 142 feral pigeons and 108 zoo animals, including samples from 28
139
carnivores, 43 ungulates, 35 birds and 2 monkeys were examined to detect antibodies
140
against T. gondii using the modified agglutination test (MAT) as previously described
141
[17]. Serum was tested at 1:25, 1:50, 1:100 and 1:500 dilutions and both positive and
142
negative controls were included in all tests. Sera with titers ≥ 1:25 were considered
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positive.
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2.3 Statistical analysis
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The prevalence of the different pathogens was estimated from the ratio of
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positive to the total number of samples tested, with the exact binomial confidence
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intervals of 95% (CI95%). Correlation between estimated prevalences and independent
148
variables (species, sex and age) were analyzed by means of a Pearson’s chi-square test
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or, when there were less than six observations per category, by the Fisher´s exact test. The
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differences between variables were analyzed by Tukey tests. Values with P < 0.05 were
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considered as statistically significant. Statistical analyses were performed using SPSS
152
22.0 (Statistical Package for Social Sciences (SPSS) Inc., Chicago, IL, USA).
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3. Results Antibodies against flavivirus of the JEV serocomplex were found in 11 of 142
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pigeons (7.8%; CI95%: 3.7-11.8) tested by bELISA. No statistically significant
158
differences were observed among sex or age. Six of the 11 positive samples were
159
confirmed by VNT (Table 2). Specific antibodies against WNV were detected in three
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pigeons (3/142: 2.1%; CI95%: 0.0-4.3), while USUV infection was also confirmed in
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three individuals (3/142: 2.1%; CI95%: 0.0-4.3). No MBV-specific antibodies were
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detected in the pigeon sera tested.
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Seroprevalence of flaviviruses in zoo animals was 8.2% (4/49; CI95%: 0.9-15.4).
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Presence of antibodies was found in ostrich (Struthio camelos), white stork (Ciconia
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ciconia), emperor goose (Chen canagica) and black wildebeest (Connochaetes gnou)
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(Table 2). Specific antibodies against WNV (1:40) and USUV (1:20) were confirmed in
167
white stork and ostrich, respectively. Moreover, the seropositive emperor goose found
168
positive in bELISA presented neutralizing antibodies against WNV (1:80), USUV
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(1:40) and MBV (1:20), which are indicative of at least two independent flavivirus
170
infections (by a flavivirus belonging to the JEV serocomplex like WNV or USUV and
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tick-borne flavivirus like MBV).
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Antibodies against AIV were not found in any of the 148 pigeons tested (0.0%;
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CI95%: 0.0-1.9). Seropositivity was observed in five out of 28 (17.9%; CI95%: 3.7-32.0)
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sera from captive zoo birds, including mallard (Anas platyrhynchos), emperor goose,
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African sacred ibis (Threskiornis aethiopicus), rosy-billed pochard (Netta peposaca)
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and black-bellied whistling duck (Dendrocygna autummalis) (Table 1).
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Salmonella spp. was not isolated from any of the 152 (0.0%; 95% CI: 0.0-1.9)
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pigeons analyzed. In contrast, in three faecal samples from the 44 captive zoo 8 Page 8 of 21
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animals tested (6.8%) positive isolation results were obtained (Table 1).
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In particular, Salmonella Panama was detected in wild boar (Sus scrofa), while
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Salmonella Apapa was isolated in milk snake (Lampropeltis triangulum). The
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Salmonella serovar isolated in ostrich could not be identified. Antibodies against T. gondii were observed in 9.2% of 142 (CI95%: 4.8-13.6)
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feral pigeons. Titers of 1:25, 1:50, 1:100 and 1:500 were detected in three (23.1%), six
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(46.2%), three (23.1%) and one (7.7%) pigeons, of the 13 seropositive animals,
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respectively. Statistically significant differences between the seropositivity against T.
187
gondii and sex or age were not found. The overall seropositivity to T. gondii in zoo
188
animals was 26.9% (29/108; CI95%: 19.6-34.1). Antibodies were detected in 42.9%
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carnivore (12/28), 30.2% ungulate (13/43) and 11.4% bird (4/35) samples, but were not
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detected in two primates analyzed. Titers of 1:25 were found in nine (31.0%), 1:50 in
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three (10.3%), 1:100 in 10 (34.5%) and 1:500 in seven (24.1%) of the 29 seropositive
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animals. Statistically significant differences between seropositivity and sex, age or
193
group of species (birds, carnivores, ungulates or primates) were not observed.
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4. Discussion
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The results from the present study confirm local circulation of flavivirus,
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including WNV and USUV, in non-migratory feral pigeons in Córdoba city. Five out of
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11 seropositive pigeons detected by bELISA were negative in USUV, WNV and MBV
198
VNT. The results could be due to the higher sensitivity of the bELISA test, to the
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detection of different types of antibodies using bELISA and VNTs, or to the circulation
200
of other antigenically related flaviviruses [18]. Accordingly, circulation and mortality
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associated to Bagaza virus infections have been detected in wild bird species, including
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the genus Columba, in Spain [19,20]. Similar rates of seropositivity against WNV and 9 Page 9 of 21
USUV were detected in pigeons and zoo birds, which indicates a homogeneous
204
circulation of these flaviviruses in these species. To date, the WNV outbreaks reported
205
in horses (n= 88) and in humans (n= 2) in Spain have only been detected in the southern
206
region [21]. Even though pigeons are included in the Spanish WNV Monitoring
207
Program [22], to our knowledge, these are the first results in this species in Spain.
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Similar seroprevalences against flaviviruses were previously reported in both horses and
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wild birds in southern Spain [23,24,25], which suggests an endemic circulation of
210
flaviviruses in this area. Higher seroprevalence (12.8%) to WNV was reported in game
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birds (partridges and pheasants) sampled in the same region where the highest number
212
of outbreaks in horses in Spain was reported [26]. The seroprevalence to WNV in
213
pigeons from other European countries ranges between 0 and 54% [27,14], and pigeons
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have been proposed as interesting sentinels of WNV transmission risk to humans [14].
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During the last few years, the number of USUV outbreaks reported in birds and humans
216
has significantly increased in different European countries, including Spain [28,29,30].
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Seroprevalence obtained in pigeons in this study was in accordance with those
218
previously reported in columbids and other wild bird species in Europe [26,14].
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Specific antibodies against WNV and USUV were confirmed in captive white
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stork and ostrich, respectively. Furthermore, one emperor goose showed specific
221
neutralizing antibodies against the three flaviviruses tested using VNT. Although the
222
higher titers against WNV detected in this species suggest infection with this flavivirus,
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contact with USUV and/or MBV cannot be ruled out. Interestingly, MBV circulation
224
has been detected in gull populations in Spain [15]. Susceptibility to WNV infection in
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white stork and emperor goose has been previously confirmed [31,32]. Seroprevalence
226
to USUV detected in zoo birds in our study (3.6%) was in accordance with that found in
10 Page 10 of 21
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other zoos in Europe [33]. To the best of our knowledge, this is the first report of USUV
228
infection in ostriches. None of the pigeons tested showed antibodies against AIV. Although
230
susceptibility to AIV infection in columbids has been previously demonstrated [34], our
231
results are consistent with that reported in columbid species in Spain [35] and other
232
European countries [36,37]. Experimental studies have showed that pigeons could be
233
considered as AIV “dead end” host because the shedding levels are below the threshold
234
of the minimal infective particles required to infect other species. Therefore,
235
disseminating AIV in this species is more likely to occur via mechanical route [34].
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AIV seropositivity was detected in five captive bird species, four of them belonging to
237
the order Anseriformes, which are the main reservoirs of AIV [38]. To author’s
238
knowledge, this is the first report of antibodies against AIV in African sacred ibis.
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The absence of Salmonella spp. isolation in pigeons suggests a limited role of
240
feral pigeons in the transmission of this enteropathogen in the study area. These results
241
are consistent with those previously reported in European countries including Spain
242
[10,39,40]. Salmonella spp. was isolated in 3 out of 44 samples from zoo animals, in
243
particular, in wild boar, milk snake and in ostrich. Due the intermittent shedding of
244
Salmonella spp. in feces; the prevalence found might have been underestimated. The
245
number of reports about Salmonella infections in humans associated to contact with
246
reptiles has increased in the last years due to the use of these species as exotic pets [41].
247
Also, clinical infections have been confirmed in zoo animals and zoo visitors by direct
248
or indirect contact with reptiles infected with Salmonella spp. [42,43]. The serotype
249
(Salmonella Apapa) detected in milk snake in the present study has been previously
250
associated with sepsis in human neonates due to direct or indirect contact with bearded
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dragons [44]. Salmonella Panama was confirmed in one wild boar without clinical
252
symptoms. Although this species is considered natural reservoir of Salmonella spp. in
253
Spain [45] and this serotype has previously been isolated in domestic pig [46], this is
254
the first report of Salmonella Panama infection in wild boar. Pigeons are a good model and indirectly indicate soil contamination by T. gondii
256
oocysts, since they feed from the ground and they are highly susceptible to oral
257
infection with oocysts [47]. Importantly, an association of the consumption of pigeon
258
meat with T. gondii infection has been reported [48]. There have been many reports of
259
T. gondii antibodies in feral pigeons, and the parasite has been isolated from this species
260
[48] but to our knowledge there have not been previous reports of seroprevalence of T.
261
gondii in feral pigeons in Spain. Seroprevalence levels observed in the present study
262
(less than 10%) are in agreement with most of the previously reported studies in pigeons
263
worldwide [49]. The wide geographic distribution of Columba livia, the ecological
264
association to human settlements and the sedentary behavior suggest the pigeon as good
265
sentinel species for the presence of oocysts in the human environment. Additional
266
studies are required in order to investigate the role of pigeons in the epidemiology of
267
T. gondii.
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Widespread T. gondii seroprevalence was observed in captive zoological species
269
in CMZP. Sixteen of 33 (48.5%) zoological species analyzed had at least one
270
seropositive animal to T. gondii with 26.9% as average seroprevalence. This study is the
271
first report of presence of T. gondii antibodies in European mink, a critically endangered
272
species [50]. High seroprevalence was observed in the felid species (46.2%), and
273
importantly all the samples analyzed from lions, jaguars and Bengal tigers were
274
positive. These results could be of Animal Health and Conservation concern in CMZP
12 Page 12 of 21
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due to the fact that seropositive felids might shed oocysts and the high susceptibility to
276
T. gondii infection of some zoo species such as Marsupials and New World monkeys
277
[49,51,52]. In the present study, antibodies against T. gondii were widespread in captive-
279
ungulates, being observed in 6 of 9 species analyzed. In Spain, presence of such
280
antibodies has been previously described in free-living wild ruminants [53] and in wild
281
boars [54]. Birds showed lower seroprevalence levels and antibodies were detected in
282
only 4 of 14 species analyzed. The prevalence of antibodies against T. gondii in wild
283
bird species in Spain has been highly heterogeneous, depending on their feeding
284
behaviour and geographical location with higher levels in carnivore wild birds [55].
285
Low number of carnivore birds was included in the present study, which could be one
286
reason for the low prevalence observed. Another explanation could be low exposure
287
to oocysts in the environment. In addition, some recent studies have suggested that
288
lower sensitivity of serological tests, such as MAT, in birds may sub-estimate
289
seropositivity levels in these species [56].
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The results obtained in the present study suggest a limited role of pigeons in the
291
transmission of AIVs and Salmonella spp. However, the seroprevalence against
292
flaviviruses and T. gondii found in this species, as well the presence of flaviviruses,
293
AIV, Salmonella spp. and T. gondii detected in captive zoo animals, indicate the need
294
to monitor these species in the study area. Control measures, including monitoring,
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population control, vaccination programs and management measures should be
296
maintained and/or implemented in order to prevent transmission of these zoonotic
297
agents between pigeons and sympatric species, included human.
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Acknowledgements 13 Page 13 of 21
We would like to thank the Municipal Council of Córdoba for providing
300
technical support. We thank Carlos Expósito in the samples collection, Cécile Beck and
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Steeve Lowenski in the virus neutralization tests, Maria Puig in the modified
302
agglutination tests, and Carmen Borge and Estefanía Jurado in the Salmonella
303
isolations.
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304 305
References
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[1] R. Sacchi, A. Gentilli, E. Razzetti, F. Barbieri, Effects of building features on density and flock distribution of feral pigeons Columba livia var. domestica in an urban environment, Can. J. Zool. 80 (2002) 48-54.
M
[2] J.C. Senar, J. Carrillo, L. Arroyo, T. Montalvo, V. Peracho, Estima de la abundancia de palomas (Columba livia var.) de la ciudad de Barcelona y valoración de la efectividad del control por eliminación de individuos, Arx. misc. zool. 7 (2009) 62–71.
d
[3] I. Amoruso, L. Fabbris, M. Mazza, G. Caravello, Estimation of Feral Pigeon (Columba livia) population size using a novel Superimposed Urban Strata (SUS) method, Urban Ecosyst. 17 (2014) 597-612. [4] P. Albonetti, A. Marletta, I. Repetto, E.A. Sasso, Efficacy of nicarbazin (Ovistop®) in the containment and reduction of the populations of feral pigeons (Columba livia var. domestica) in the city of Genoa, Italy: a retrospective evaluation, Vet Ital. 51 (2015) 6372.
Ac ce pt e
307 308 309 310 311 312 313 314 315 316 317 318 319 320 321 322 323 324 325 326 327 328 329 330 331 332 333 334 335 336 337 338 339 340 341
cr
306
[5] T. Mcnamara, The role of zoos in biosurveillance, Int. Zoo Yb. 41 (2007) 12-15. [6] A. Dovc, O. Zorman-Rojs, A. Vergles-Rataj, V. Bole-Hribovsek, U. Krapez, M. Dobeic, Health status of free-living pigeons (Columba livia domestica) in the city of Ljubljana, Acta Vet Hung. 52 (2004) 219-226. [7] E. De-Sousa, A.J. Berchieri, A.A. Pinto, R.Z. Machado, A.O. de Carrasco, J.A. Marciano, K. Werther, Prevalence of Salmonella spp. Antibodies to Toxoplasma gondii, and Newcatle disease virus in feral pigeons (Columba livia) in the city of Jaboticabal, Brazil, J Zoo Wildl Med. 41 (2010) 603-607. [8] J. Salinas, M.R. Caro, F. Cuello, Antibody prevalence and isolation of Chlamydia psittaci from pigeons (Columba livia), Avian Dis. 37 (1993) 523-7. [9] B. Vázquez, F. Esperon, E. Neves, J. Lopez, C. Ballesteros, M.J. Munoz, Screening for several potential pathogens in feral pigeons (Columba livia) in Madrid, Acta Vet Scand. 52 (2010) 45. 14 Page 14 of 21
[10] L. Casanovas, M. de Simón, M.D. Ferrer, J. Arqués, G. Monzón, Intestinal carriage of campylobacters, salmonellas, yersinias and listerias in pigeons in the city of Barcelona, J Appl Bacteriol. 78 (1995) 11-3. [11] M.F. Colom-Valiente, M. Alberdi, I. Meseguer, J.M. Torres-Rodríguez, Isolation of Cryptococcus neoformans from environmental samples in Alicante, Rev Iberoam Micol. 14 (1997) 63-4.
ip t
[12] B. Close, K. Banister, V. Baumans, E.M. Bernoth, N. Bromage, J. Bunyan, W. Erhardt, P. Flecknell, N. Gregory, H. Hackbarth, D. Morton, C. Warwick, Recommendations for euthanasia of experimental animals: Part 2. Lab. Lab Anim. 31 (1997) 1-32.
us
cr
[13] D. Gibbs, E. Barnes, C. John, Pigeons and Doves: A Guide to the Pigeons and Doves of the World, Christppher Helm Publishers, London, 2010.
an
[14] S.C. Chaintoutis, C.I. Dovas, M. Papanastassopoulou, S. Gewehr, K. Danis, C. Beck, S. Lecollinet, V. Antalis, S. Kalaitzopoulou, T. Panagiotopoulos, S. Mourelatos, S. Zientara, O. Papadopoulos, Evaluation of a West Nile virus surveillance and early warning system in Greece, based on domestic pigeons, Comp Immunol Microbiol Infect Dis. 37 (2014) 131-41.
d
M
[15] A. Arnal, E. Gómez-Díaz, M. Cerdà-Cuéllar, S. Lecollinet, J. Pearce-Duvet, N. Busquets, I. García-Bocanegra, N. Pagès, M. Vittecoq, A. Hammouda, B. Samraoui, R. Garnier, R. Ramos, S. Selmi, J. González-Solís, E. Jourdain, T. Boulinier, Circulation of a Meaban-like virus in yellow-legged gulls and seabird ticks in the western Mediterranean basin, PLoS One 13 (2014) e89601. [16] M.Y. Popoff, J. Bockemühl, F.W. Brenner, L.L. Gheesling, Supplement 2000 (no. 44) to the Kauffmann-White scheme, Res Microbiol. 152 (2001) 907-9.
Ac ce pt e
342 343 344 345 346 347 348 349 350 351 352 353 354 355 356 357 358 359 360 361 362 363 364 365 366 367 368 369 370 371 372 373 374 375 376 377 378 379 380 381 382 383 384 385 386 387 388 389 390 391
[17] J.P. Dubey, G. Desmonts, Serological responses of equids fed Toxoplasma gondii oocysts, Equine.Vet. J. 19(4) (1987) 337–339. [18] B.J. Blitvich, N.L. Marlenee, R.A. Hall, C.H. Calisher, R.A. Bowen, J.T. Roehring, N. Komar, S.A. Langevin, B.J. Beaty, Epitope-blocking enzyme-linked immunosorbent assays for the detection of serum antibodies to West Nile virus in multiple avian species, J Clin Microbiol. 41 (2003) 1041-7. [19] V. Gamino, A.V. Gutiérrez-Guzmán, I.G. Fernández-de-Mera, J.A. Ortíz, M. Durán-Martín, J. De la Fuente, C. Gortázar, U. Höfle, Natural Bagaza virus infection in game birds in southern Spain, Vet Res. 43 (2012) 65. [20] I. García-Bocanegra, I. Zorrilla, E. Rodríguez, E. Rayas, L. Camacho, I. Redondo, F. Gómez-Guillamón, Monitoring of the Bagaza virus epidemic in wild birds species in Spain, 2010, Transbound Emerg Dis. 60 (2013) 120-6. [21] RASVE (Red de Alerta Sanitaria Veterinaria), 2015. Available at: http://rasve.magrama.es/RASVE_2008/Publica/Focos/Consulta.aspx (accessed April 15, 2015). 15 Page 15 of 21
[22] RASVE (Red de Alerta Sanitaria Veterinaria), 2015. Available at: http://rasve.magrama.es/Publica/Programas/Normativa.asp (accessed April 25, 2015). [23] I. García-Bocanegra, N. Busquets, S. Napp, A. Alba, I. Zorrilla, R. Villalba, A. Arenas, Serosurvey of West Nile Virus and Other Flaviviruses of the Japanese Encephalitis Antigenic Complex in Birds from Andalusia, Southern Spain, Vector Borne Zoonotic Dis. 11 (2011) 1107-1113.
ip t
[24] I. García-Bocanegra, J.A. Jaén-Téllez, S. Napp, A. Arenas-Montes, M. FernándezMorente, V. Fernández-Molera, A. Arenas, Monitoring of the West Nile virus epidemic in Spain between 2010 and 2011, Transbound Emerg Dis. 59 (2012) 448-55.
us
cr
[25] G. López, M.A. Jiménez-Clavero, A. Vázquez, R. Soriguer, C. Gómez-Tejedor, A. Tenorio, J. Figuerola, Incidence of West Nile virus in birds arriving in wildlife rehabilitation centers in southern Spain, Vector Borne Zoonotic Dis. 11 (2011) 285-90.
an
[26] F. Llorente, E. Pérez-Ramírez, J. Fernández-Pinero, R. Soriguer, J. Figuerola, M.A. Jiménez-Clavero, Flaviviruses in game birds, southern Spain, 2011-2012, Emerg Infect Dis. 19 (2013) 023-5.
M
[27] P. Lena, E. Bureau, S. Reynard, P. Sabatier, S. Bruyas, H. Zeller, Sero-survey for West Nile in wild birds in "The Dombes" Wetlands (01, Ain. France) in 2001: absence of detection of viral activity, Rev Med Vet-Toulouse. 157 (2006) 612-618.
d
[28] H. Weissenböck, J. Kolodziejek, A. Url, H. Lussy, B. Rebel-Bauder, N. Nowotnyz, Emergence of Usutu virus, an African mosquito-borne flavivirus of the Japanese encephalitis virus group, central Europe, Emerg Infect Dis. 8 (2002) 652-6.
Ac ce pt e
392 393 394 395 396 397 398 399 400 401 402 403 404 405 406 407 408 409 410 411 412 413 414 415 416 417 418 419 420 421 422 423 424 425 426 427 428 429 430 431 432 433 434 435 436 437 438 439 440 441
[29] U. Höfle, V. Gamino, I.G. Fernández de Mera, A.J. Mangold, J.A. Ortíz, J. de la Fuente, Usutu Virus in Migratory Song Thrushes, Spain, Emerg Infect Dis. 19 (2013) 1023–1025. [30] M. Santini, T. Vilibic-Cavlek, B. Barsic, L. Barbic, V. Savic, V. Stevanovic, E. Listes, A. Di Gennaro, G. Savini, First cases of human Usutu virus neuroinvasive infection in Croatia, August-September 2013: clinical and laboratory features, J Neurovirol. 21 (2015) 92-7. [31] M. Malkinson, Y. Weisman, S. Pokamonski, R. King, V. Deubel, Intercontinental transmission of West Nile virus by migrating white storks, Emerg Infect Dis. 7 (2001) 540. [32] CDC (Centers for Diseases Control and Prevention), 2015. Available at: http://www.cdc.gov/westnile/resources/pdfs/Bird%20Species%201999-2012.pdf (accessed May 10, 2015). [33] N. Buchebner, W. Zenker, C. Wenker, H.W. Steinmetz, E. Sós, N. Nowotny, Low Usutu virus seroprevalence in four zoological gardens in central Europe, BMC Vet Res. 9 (2013) 153. [34] C. Abolnik, A current review of avian influenza in pigeons and doves (Columbidae), Vet Microbiol. 170 (2014) 181–196. 16 Page 16 of 21
[35] E. Pérez-Ramírez, X. Gerrikagoitia, M. Barral, U. Höfle, Detection of low pathogenic avian influenza viruses in wild birds in Castilla-La Mancha (south central Spain), Vet Microbiol. 146 (2010) 200-208.
ip t
[36] A. Lillehaug, C. Monceyron-Jonassen, B. Bergsjø, M. Hofshagen, J. Tharaldsen, L.L. Nesse, K. Handeland, Screening of feral pigeon (Columba livia), mallard (Anas platyrhynchos) and graylag goose (Anser anser) populations for Campylobacter spp., Salmonella spp., avian influenza virus and avian paramyxovirus, Acta Vet Scand. 46 (2005) 193-202.
cr
[37] L. Teske, M. Ryll, S. Rautenschlein, Epidemiological investigations on the role of clinically healthy racing pigeons as a reservoir for avian paramyxovirus-1 and avian influenza virus, Avian Pathol. 42 (2013) 557-65.
us
[38] J.K. Kim, N.J. Negovetich, H.L. Forrest, R.G. Webster, Ducks: the “Trojan horses” of H5N1 influenza, Influenza Other Resp. 3 (2009) 121-8.
an
[39] L. Teske, M. Ryll, D. Rubbenstroth, I. Hänel, M. Hartmann, L. Krejenbrock, S. Rautenschlein. Epidemiological investigations on the possible risk of distribution of zoonotic bacteria through apparently healthy homing pigeons, Avian Pathol. 42 (2013) 397-407.
d
M
[40] A. Gargiulo, T.P. Russo, R. Schettini, K. Mallardo, M. Calabria, L.F. Menna, P. Raia, U. Pagnini, V. Caputo, A. Fioretti, L. Dipineto, Occurrence of enteropathogenic bacteria in urban pigeons (Columba livia) in Italy, Vector Borne Zoonotic Dis. 14 (2014) 251-5. [41] L. Geue, U. Löschner, Salmonella enterica in reptiles of German and Austrian origin, Vet Microbiol. 84 (2002) 79–91.
Ac ce pt e
442 443 444 445 446 447 448 449 450 451 452 453 454 455 456 457 458 459 460 461 462 463 464 465 466 467 468 469 470 471 472 473 474 475 476 477 478 479 480 481 482 483 484 485 486 487 488 489 490 491
[42] C.R. Friedman, C. Torigian, P.J. Shillam, R.E. Hoffman, D. Heltzel, J.L. Beebe, G. Malcolm, W.E. Dewitt, L. Hutwagner, P.M. Griffin, An outbreak of salmonellosis among children attending a reptile exhibit at a zoo, J Pediatr. 132 (1998) 802–807. [43] L. Bauwens, F. Vercammen, S. Bertrand, J.M. Collard, S. De Ceuster, Isolation of Salmonella from environmental samples collected in the reptile department of Antwerp Zoo using different selective methods, J Appl Microbiol. 101 (2006) 284–289. [44] R. Haase, T. Beier, M. Bernstadt, N. Merkel, J. Bartnicki, Neonatal infection with Salmonella apapa after contact with a reptile in the home, Z. Geburtsh Neonatol. 215 (2011) 86-88. [45] N. Navarro-González, G. Mentaberre, C.M. Porrero, E. Serrano, A. Mateos, J.M. López-Martín, S. Lavín, L. Domínguez, Effect of Cattle on Salmonella Carriage, Diversity and Antimicrobial Resistance in Free-Ranging Wild Boar (Sus scrofa) in Northeastern Spain, PLoS One 7 (2012) e51614. [46] J.D. Kich, A. Coldebella, N. Mores, M. Nogueira, M. Cardoso, P.M. Fratamico, J.E. Call, P. Fedorka-Cray, J.B. Luchansky, Prevalence, distribution, and molecular 17 Page 17 of 21
characterization of Salmonella recovered from swine finishing herds and a slaughter facility in Santa Catarina, Brazil, Int J Food Microbiol. 151 (2011) 307-13. [47] H. Waap, R. Cardoso, A. Leitão, T. Nunes, A. Vilares, M.J. Gargaté, J. Meireles, H. Cortes, H. Ângelo, In vitro isolation and seroprevalence of Toxoplasma gondii in stray cats and pigeons in Lisbon, Portugal, Vet Parasitol. 187 (2012) 542-7.
cr
ip t
[48] C. Alvarado-Esquivel, O. Liesenfeld, B.D. Burciaga-López, A. Ramos-Nevárez, S. Estrada-Martínez, S.M. Cerrillo-Soto, F.A. Carrete-Ramírez, Mde.L. López-Centeno, M.M. Ruiz-Martínez, Seroepidemiology of Toxoplasma gondii infection in elderly people in a northern Mexican city, Vector Borne Zoonotic Dis. 12 (2012) 568-74. [49] J.P. Dubey, Toxoplasmosis in animals and humans, 2nd ed., CRC Press, Boca Raton, FL, 2010.
us
[50] UICN (International Union for Conservation of Nature), 2015. Available at: http://www.iucnredlist.org/details/14018/0 (accessed April 5, 2015).
an
[51] C. Juan-Sallés, N. Prats, A.J. Marco, J.A. Ramos-Vara, D. Borrás, J. Fernández, Fatal acute toxoplasmosis in three golden lion tamarins (Leontopithecus rosalia), J Zoo Wildl Med. 29 (1998) 55-60.
M
[52] X. Fernández-Aguilar, D. Ajzenberg, O. Cabezón, A. Martínez-López, L. Darwich, J.P. Dubey, S. Almería, Fatal toxoplasmosis associated with an atypical Toxoplasma gondii strain in a Bennett's wallaby (Macropus rufogriseus) in Spain, Vet Parasitol. 196 (2013) 523-527.
d
[53] C.B. Gauss, J.P. Dubey, D. Vidal, O. Cabezón, F. Ruiz-Fons, J. Vicente, I. Marco, S. Lavin, C. Gortazar, S. Almería, Prevalence of Toxoplasma gondii antibodies in red deer (Cervus elaphus) and other wild ruminants from Spain, Vet Parasitol. 136 (2006) 193-200.
Ac ce pt e
492 493 494 495 496 497 498 499 500 501 502 503 504 505 506 507 508 509 510 511 512 513 514 515 516 517 518 519 520 521 522 523 524 525 526 527 528 529 530 531 532 533 534 535 536 537 538
[54] C.B. Gauss, J.P. Dubey, D. Vidal, F. Ruiz, J. Vicente, I. Marco, S. Lavin, C. Gortazar, S. Almería, Seroprevalence of Toxoplasma gondii in wild pigs (Sus scrofa) from Spain, Vet Parasitol. 131 (2005) 151-6. [55] O. Cabezón, I. García-Bocanegra, R. Molina-López, I. Marco, J.M. Blanco, U. Höfle, A. Margalida, E. Bach-Raich, L. Darwich, I. Echeverría, E. Obón, M. Hernández, S. Lavín, J.P. Dubey, S. Almería, Seropositivity and risk factors associated with Toxoplasma gondii infection in wild birds from Spain, PLoS One 6 (2011) e29549. [56] L. Casartelli-Alves, V.C. Boechat, R. Macedo-Couto, L.C. Ferreira, J.L. Nicolau, L.B. Neves, P.R. Millar, R.T. Vicente, R.V. Oliveira, A.G. Muniz, I.C. Bonna, M.R. Amendoeira, R.C. Silva, H. Langoni, T.M. Schubach, R.C. Menezes, Sensitivity and specificity of serological tests, histopathology and immunohistochemistry for detection of Toxoplasma gondii infection in domestic chickens, Vet Parasitol. 204 (2014) 34651.
18 Page 18 of 21
Tables
539
Table 1. Prevalence of selected zoonotic pathogens in feral pigeons (Columba livia var.
540
domestica) and sympatric zoological species in Córdoba, Southern Spain.
Ac ce pt e
d
M
an
us
cr
ip t
538
19 Page 19 of 21
Salmonella spp.
Toxoplasma gondii
0/148
0/152
13/142 (9.2%)
0/3 0/7 0/1 0/5 0/3 1/1 (100 %) 1/20 (5%)
-
0/1 0/1 0/1 0/1 1/1 (100 %) 0/1 1/6 (16.7%)
2/10 (20%) 3/15 (20%) 1/1(100%) 4/6 (66.7%) 2/3 (66.7%) 0/1 1/3 (33.3%) 0/3 13/42 (31%)
0/1 0/1
-
0/3 0/1 0/4
0/1 0/1
0/1 0/3 0/4 0/3 1/2 (50%) 1/13 (7.7%)
1/1 (100 %) 1/3 (33.3 %) 1/4 (25 %) 0/3 1/2 (50 %) 4/13 (30.8%)
0/1 0/1 0/2
0/1 0/3 1/4 (25 %) 1/11 (9.1%) 1/2 (50%) 3/21 (14.3%)
0/1 0/1 0/2
0/1 0/1 0/1 0/1 0/4
0/1 1/1 (100%) 1/2 (50%)
0/1
0/1
0/1
0/1
0/2 0/4 1/2 (50 %) 1/8 (12.5%)
0/2 0/4 0/2 0/8
0/1 0/1 1/2 1/4 (25%)
0/2 0/4 0/2 0/8
1/2 (50 %)
0/2
-
0/1
0/2
1/2 (50 %)
-
0/1
-
-
-
0/1
-
-
0/1 0/2 0/1 0/1 0/5
2/2 (100%) 1/1 (100%) 3/3 (100%) 0/7 3/3 (100%) 0/1 1/8 (12.5%) 2/3 (66.6%) 12/28 (42.9%)
-
-
0/2 0/1 0/1 0/1 0/2 0/2 0/2 0/1 0/1 0/1 0/14
0/1 0/1 0/2
-
-
0/1 0/1 0/2
-
-
-
0/1 1/1 (100 %) 1/2 (50 %)
4/49 (8.2%)
5/28 (17.9 %)
0/2 3/44 (6.8%)
M
us
-
an
11/142 (7.8 %)
ip t
Influenza
cr
Flaviviruses
Ac ce pt e
d
SPECIES ORDER COLUMBIFORMES Feral pigeon (Columba livia var. domestica) ORDER ARTIODACTYLA Aoudad (Ammotragus lervia) Red deer (Cervus elaphus) Fallow deer (Dama dama) Mouflon (Ovis musimon) Collared Peccary (Pecari tajacu) Black Wildebeest (Connochaetes gnou) Roe deer (Capreolus capreolus) Wild boar (Sus scrofa) Llama (Lama glama) Sheep (Ovis aries) Goat (Capra hircus) TOTAL ORDER PERISSODACTYLA Plains zebra (Equus burchellii) Donkey (Equus africanus asinus) Lowland Tapir (Tapirus terrestris) TOTAL ORDER ANSERIFORMES Rosy-billed Pochard (Netta peposaca) Black-bellied whistling duck (Dendrocygna autummalis) Mallard (Anas platyrhynchos) Bar-headed goose (Anser indicus) Emperor goose (Chen canagica) TOTAL ORDER ACCIPITRIFORMES Griffon vulture (Gyps fulvus) Booted eagle (Hieraaetus pennatus) Short-toed snake eagle (Circaetus gallicus) Common buzzard (Buteo buteo) TOTAL ORDER GALLIFORMES Indian peafowl (Pavo cristatus) ORDER STRUTHIONIFORMES Greater rhea (Rhea americana) Emu (Dromaius novaehollandiae) Ostrich (Struthio camelus) TOTAL ORDER CICONIIFORMES White stork (Ciconia ciconia) ORDER PELECANIFORMES African sacred ibis (Threskiornis aethiopicus) ORDER PSITTACIFORMES Muir's corella (Cacatua pastinator) ORDER CARNIVORA Lion (Panthera leo) Jaguar (Panthera onca) Bengal tiger (Panthera tigris tigris) Eurasian lynx (Lynx lynx) Brown bear (Ursus arctos pyrenaicus) European otter (Lutra lutra) Iberian wolf (Canis lupus signatus) European mink (Mustela lutreola) TOTAL ORDER PRIMATES Northern white-cheeked gibbon (Hylobates leucogenys) Brown lemur (Eulemur fulvus) Black-and-white ruffed lemur (Varecia variegata) Ring-tailed lemur (Lemur catta) Barbary macaque (Macaca sylvanus) White-collared Mangabey (Cercocebus atys ssp. lunulatus) De Brazza’s Monkey (Cercopithecus neglectus) White-tufted-ear Marmoset (Callithrix jacchus) Western Pygmy Marmoset (Callithrix pygmaea) South American Squirrel Monkey (Saimiri sciureus) TOTAL ORDER RODENTIA Patagonian Cavy (Dolichotis patagonum) Capybara (Hydrochoerus hydrochaeris) TOTAL ORDER SQUAMATA Corn snake (Elaphe guttata) Milk snake (Lampropeltis triangulum) TOTAL ORDER TESTUDINES African spurred tortoise (Geochelone sulcata) TOTAL IN CAPTIVE ANIMALS
-20 -
Page 20 of 21
29/108 (26.9%)
541 542
Table 2. Results of virus neutralization test for the detection of flaviviruses in feral
543
pigeons and captive zoo animals. Species, (no.)
VNT titers WNV
USUV
MBV
Feral pigeon (n= 1)
<10
10
<20
Feral pigeon (n= 1)
20
<10
<20
Feral pigeon (n= 1)
80
<10
<20
WNV
Feral pigeon (n= 1)
<10
10
<20
USUV
Feral pigeon (n= 1)
80
10
<20
WNV
Feral pigeon (n= 1)
<10
20-40
<20
USUV
Feral pigeon (n= 5)
<10
Total (%)
3(2.1)
ip t
Feral pigeon (Columba livia) (n=142)
USUV
an
us
cr
WNV
<20
3(2.1)
0
Undetermined Flavivirus
M
<10
Emperor goose (Chen canagica) (n= 1)
80
40
20
Undetermined Flavivirus
White stork (Ciconia ciconia) (n= 1)
40
<10
NA1
WNV
<10
20
<20
USUV
<10
<10
<20
Undetermined Flavivirus
1(2.0)
1(2.0)
Ac ce pt e
Ostrich (Rhea americana) (n= 1)
d
Zoo animals (n=49)
Black wildebeest (Connochaetes gnou) (n= 1) Total (%)
544 545
Interpretation
WNV, West Nile virus; USUV, Usutu virus; MV, Meaban virus; VNT, virus neutralization test; 1. NA: Not analyzed, low volume.
21 Page 21 of 21