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High Prevalence of Antibodies Against Chlamydiaceae, Chlamydophila Abortus in Wild Ungulates Using Two “In House” Blocking-ELISA Tests J

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High Prevalence of Antibodies Against Chlamydiaceae, Chlamydophila Abortus in Wild Ungulates Using Two “In House” Blocking-ELISA Tests J High prevalence of antibodies against Chlamydiaceae, Chlamydophila abortus in wild ungulates using two “in house” blocking-ELISA tests J. Salinas, M.R. Caro, J. Vicente, F. Cuello, A.R. Reyes-Garcia, A.J. Buendía, A. Rodolakis, C. Gortázar To cite this version: J. Salinas, M.R. Caro, J. Vicente, F. Cuello, A.R. Reyes-Garcia, et al.. High prevalence of antibodies against Chlamydiaceae, Chlamydophila abortus in wild ungulates using two “in house” blocking- ELISA tests. Veterinary Microbiology, Elsevier, 2009, 135 (1-2), pp.46. 10.1016/j.vetmic.2008.10.001. hal-00532520 HAL Id: hal-00532520 https://hal.archives-ouvertes.fr/hal-00532520 Submitted on 4 Nov 2010 HAL is a multi-disciplinary open access L’archive ouverte pluridisciplinaire HAL, est archive for the deposit and dissemination of sci- destinée au dépôt et à la diffusion de documents entific research documents, whether they are pub- scientifiques de niveau recherche, publiés ou non, lished or not. The documents may come from émanant des établissements d’enseignement et de teaching and research institutions in France or recherche français ou étrangers, des laboratoires abroad, or from public or private research centers. publics ou privés. Accepted Manuscript Title: High prevalence of antibodies against Chlamydiaceae, Chlamydophila abortus in wild ungulates using two “in house” blocking-ELISA tests Authors: J. Salinas, M.R. Caro, J. Vicente, F. Cuello, A.R. Reyes-Garcia, A.J. Buend´ıa, A. Rodolakis, C. Gortazar´ PII: S0378-1135(08)00479-3 DOI: doi:10.1016/j.vetmic.2008.10.001 Reference: VETMIC 4232 To appear in: VETMIC Please cite this article as: Salinas, J., Caro, M.R., Vicente, J., Cuello, F., Reyes-Garcia, A.R., Buend´ıa, A.J., Rodolakis, A., Gortazar,´ C., High prevalence of antibodies against Chlamydiaceae, Chlamydophila abortus in wild ungulates using two “in house” blocking-ELISA tests, Veterinary Microbiology (2008), doi:10.1016/j.vetmic.2008.10.001 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 High prevalence of antibodies against Chlamydiaceae and Chlamydophila abortus 2 in wild ungulates using two “in house” blocking-ELISA tests 3 4 5 J. Salinas a,* , M.R. Caro a, J. Vicente b, F. Cuello a, A.R. Reyes-Garcia b, A.J. Buendía a, A. 6 Rodolakis c, C. Gortázar b,* 7 8 a Departamento de Sanidad Animal, Facultad de Veterinaria, Universidad de Murcia, 9 Campus de Espinardo, 30100 Murcia, Spain. 10 b IREC (CSIC – UCLM – JCCM). Ronda de Toledo s.n. 13071 Ciudad Real, Spain. 11 c INRA, UR1282, Infectiologie Animale et Santé Publique, Centre de recherche de 12 Tours, 37380 Nouzilly, France. 13 14 15 16 17 * Corresponding authors: J. Salinas ( [email protected] ) 18 C. Gortázar ( [email protected] ) 19 20 21 Accepted Manuscript 22 23 24 25 1 Page 1 of 27 26 Abstract 27 Few data are available on the prevalence and relevance of chlamydiae in wild mammals, 28 and even fewer studies have been conducted to determine the prevalence of 29 Chlamydophila abortus in wildlife hosts, most probably due to the absence of suitable 30 species-specific serological assays for testing sera from wild animals. In light of this, we 31 have developed two in-house blocking-ELISA tests for detection of antibodies against 32 Chlamydiaceae and C. abortus in wild ungulates, and analyzed the relationship between 33 geographical and biological factors and the prevalence of antibodies against 34 Chlamydiaceae and C. abortus in 434 wild ungulates from Spain, including sera from 35 European wild boar, Red deer, Fallow deer, Roe deer, Mouflon, Barbary sheep, 36 Southern chamois, and Iberian ibex. Serology revealed that 41.7 ± 4% of the sera were 37 positive for the b-ELISA-LPS ( Chlamydiaceae -specific) and 18.9 ± 3% for the b- 38 ELISA-rPOMP ( C. abortus -specific). Antibodies against Chlamydiaceae LPS were 39 detected in sera from all eight ungulate species, the prevalence ranging from 23 to 60%. 40 Iberian ibex was the only wild ungulate not showing seropositivity to the C. abortus 41 specific POMP. The prevalence of anti-POMP antibodies in the other seven wild 42 ungulate species ranged from 7 to 40%. While significant seroprevalence differences 43 were detected among species and among sampling regions, no effect of age and sex was 44 observed. The high prevalence levels found should be considered with regards to 45 livestock and human health, and warrant further research. 46 Accepted Manuscript 47 Key words: Chlamydiaceae ; Chlamydophila abortus ; Blocking-ELISA; Ammotragus 48 lervia ; Capra pyrenaica ; Capreolus capreolus ; Cervus elaphus ; Dama dama ; Ovis 49 aries ; Rupicapra pyrenaica ; Sus scrofa ; Wildlife reservoir. 50 2 Page 2 of 27 51 1. Introduction 52 The link between wildlife, environment, pathogens and human activities creates 53 a dynamic where new pathogens or new hosts frequently emerge; changes in population 54 density or host behaviour affect disease prevalence, and disease agents can suddenly 55 boost their virulence and widen their host range (Gortázar et al., 2007). There is 56 growing evidence that chlamydial infections may result in adverse pregnancy outcomes 57 in humans and domestic animals (Pospischil et al., 2002; Baud et al., 2008), and it is 58 well known that chlamydiosis is a multi-host disease affecting a huge variety of 59 vertebrates (Everett et al., 1999). 60 Chlamydophila, which includes six species, is a bacterial genus that, together 61 with the genus Chlamydia, both belongs to the family Chlamydiaceae . Ovine enzootic 62 abortion (OEA) caused by Chlamydophila abortus is the most common cause of 63 reproductive failure in sheep- and goat-breeding countries in Europe, and where it has a 64 susbtantial economic impact (Aitken, 1993). Infected pregnant ewes and goats abort late 65 in gestation or give birth to weak lambs as a result of affected placenta. Bacteria 66 excreted at abortion with the uterine discharge are the main source of infection for 67 susceptible animals through ingestion or inhalation. After the onset of abortion, most 68 animals acquire immunity and rebreed successfully (Rodolakis et al., 1998). Data from 69 the few existing reports on chlamydial seroprevalence in domestic ruminants show 70 geographical differences, ranging from 11 to 51% in sheep and goat flocks (Cuello et 71 al., 1992; AcceptedMarkey et al., 1993; Mainar-Jaime etManuscript al., 1998; Borel et al., 2004) and from 72 24 to 45% in cattle (Cavirani et al., 2001; Petit et al., 2008). Data related to the 73 prevalence and relevance of chlamydial infections in wild mammals are even more 74 scarce (Hotzel et al., 2004). 3 Page 3 of 27 75 Early diagnosis of chlamydial infections is important to prevent and limit the 76 spread of the microorganism. For diagnosis purposes, serological assays are more 77 suitable for screening large numbers of samples. The complement fixation test (CFT) is 78 the most widely used test and is still recommended by the Organisation Internationale 79 des Epizooties (OIE) (http://www.oie.int). However, it lacks specificity due to its 80 antigen, which consists mainly of the heat-resistant lipopolysaccharide (LPS), common 81 to all Chlamydiaceae species (Brade et al., 1987), particularly in ruminants, which are 82 widely infected by Chlamydophila pecorum (Rodolakis et al., 1998). Moreover, CFT 83 requires a prime quality serum which, in wildlife studies, is not always available 84 (Mason and Fleming, 1999). Several ELISA methods have been developed in order to 85 improve chlamydial serology. These include ELISA tests using purified whole 86 elementary bodies (EB), LPS or more specific assays based on the C. abortus major 87 outer membrane protein (MOMP) (Kaltenboeck et al., 1997; Salti-Montesanto et al., 88 1997; Hoelzle et al., 2004; Livingstone et al., 2005). Comparative serological studies 89 have revealed that a new commercially available ELISA test based on a polymorphic 90 outer membrane protein (POMP) could serve as an improved alternative to CFT or to 91 classical indirect ELISAs for the serological diagnosis of OEA (Buendia et al., 2001; 92 Vretou et al., 2007). 93 Although there are numerous indications that Chlamydiaceae are present in a 94 wide range of wildlife host species, few studies have been conducted to determine the 95 prevalenceAccepted of C. abortus , most probably due to Manuscriptthe absence of suitable species-specific 96 serological tools and reagents to test sera from wild animals. While the role of 97 Chlamydiaceae as pathogens in European wild boar ( Sus scrofa ) has yet to be 98 established, the detection by PCR of a spectrum of Chlamydiaceae species identical to 99 those in domestic pigs (including C. abortus ) in a small sample of hunter-harvested wild 4 Page 4 of 27 100 boar from Germany, revealed a possible wildlife reservoir for these bacteria. The 101 possibility that these animals acquired chlamydiae through contact with domestic pigs 102 appeared very unlikely (Hotzel et al., 2004). 103 As regards wild ruminants, the literature on abortions caused by C. abortus 104 includes a case report on a springbok ( Antidorcas marsupialis ) in Paris zoo (Berri et al., 105 2004). However, most studies have been made by serological analysis using CFT. In 106 Italy, Giovannini et al. (1988) showed a prevalence of 79% in fallow deer ( Dama 107 dama ), while Giacometti et al. (1995) showed a prevalence of 31% in alpine ibex 108 (Capra ibex ). Similarly, a CFT-based study in populations of wild ruminants including 109 fallow deer, red deer ( Cervus elaphus ), mouflon ( Ovis aries musimon ) and Iberian ibex 110 (Capra pyrenaica ) in southern Spain indicated that Chlamydiaceae infections could be 111 present in wild ruminant species (Cubero-Pablo et al., 2000).
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