Coinfection is an important factor in epidemiological studies: the first serosurvey of the aoudad (Ammotragus lervia).

Despite being considered an invasive ungulate outside its native range (North Africa), little information exists regarding the role of the aoudad (also called Barbary sheep, Ammotragus lervia) as a pathogen reservoir. Furthermore, in most epidemiological surveys the potential role of coinfections (e.g. a first infection may make the host more immuno-competent or susceptible against a second pathogen) as a risk factor is often neglected. In this study we first performed a serological survey for selected pathogens (Mycobacterium bovis, M. avium subsp. paratuberculosis, Chlamydophila abortus, bovine viral diarrhoea/border disease viruses (BVDV-BDV), Salmonella spp., Brucella melitensis and Toxoplasma gondii) on free (n = 66) and captive (n = 25) aoudad from south-east Spain. Then, by using Akaike's information criterion, we evaluated the importance of coinfection in two statistical models that included the effects of population, age, and sex. Our results show that neither free nor captive aoudad had antibodies against Brucella melitensis, Chlamydophila abortus, or BVDV-BDV. However, compared to other wild ungulates in Spain, aoudads have high prevalence of antibodies against M. bovis (free = 49.5%; captive = 8%), very high prevalence of antibodies against M. avium subsp. paratuberculosis (free = 19.4%; captive = 56%), and intermediate prevalence of antibodies against Salmonella spp. (free = 13.4%; captive = 0%) or T. gondii (free = 1.5%; captive = 24%). Although the additive effects of population and age were included in our set of selected models, coinfection was the most influential factor to detect antibodies response against mycobacterials and salmonella infections. The direction of this influence could be exclusion of disease between tuberculosis and paratuberculosis seroreactor animals, or enhanced susceptibility to disease between tuberculosis and salmonella seroreactor animals. In conclusion, we believe that wildlife managers must pay more attention to the potential risk posed by aoudads as hosts (and probably reservoirs) of paratuberculosis and tuberculosis mycobacterials, while epidemiologists should be more aware of coinfection as an important factor in epidemiological surveys, especially in wildlife populations where multiple infections are common.

Epidemiology of Mycoplasma agalactiae infection in free-ranging Spanish ibex (Capra pyrenaica) in Andalusia, southern Spain.

Mycoplasma agalactiae is the main causal agent of contagious agalactia syndrome in Spain. It is a severe disease of small ruminants, endemic in Mediterranean countries, that is characterized by mastitis, arthritis, and keratoconjunctivitis. This paper investigates the temporal, spatial, and host-related factors in the distribution of M. agalactiae infection from October 1996 to November 1998 and March 2002 to May 2003 in Spanish ibex (Capra pyrenaica) populations from Andalusia, in southern Spain. The predisposing factors to infection among previously selected factors (year of sampling, climatic season, geographic origin according to province, mountain range and metapopulation, sex, year of life, presence of scabies, and phase of the reproductive cycle) were established. We collected conjunctival and ear-canal swabs from 411 free-ranging ibexes. The frequency of infected ibexes was 11.2%. The peak frequency of infection occurred in 1998 and in summer. Granada was the province with greatest risk (odds ratio = 2.6) of carriers (18.8% infected). The predisposing factors were sex (females), age (young animals), and metapopulation (Sierra Nevada). We identified a higher number of infected ibexes in the metapopulation "Sierra Nevada" (34/ 256) and significant differences among the three established metapopulations (P<0.01). Mycoplasma agalactiae infection represents a risk for population density and maintenance of these wild populations; infections can result in blindness, malnutrition, and polyarthritis leading to numerous deaths.

Antibodies to Mycobacterium bovis in wild carnivores from Doñana National Park (Spain).

We conducted a retrospective serologic survey for antibodies against the MPB70 protein of Mycobacterium bovis in wild carnivores from Doñana National Park (southwestern Spain). Serum samples from 118 red foxes (Vulpes vulpes), 39 Iberian lynx (Lynx pardinus), 31 Eurasian badgers (Meles meles), five Egyptian mongoose (Herpestes ichneumon), four European genet (Genetta genetta), and one Eurasian otter (Lutra lutra) were analyzed using an indirect competitive enzyme-linked immunoassay. Antibodies against the MPB70 protein of M. bovis were detected in seven badgers, five foxes, and one lynx. The frequency of positive animals was significantly higher in badger (23%) than in lynx (3%) and fox (4%). Antibodies were not detected in other species. Annual antibody frequency peaked at 38% in badgers and 11% for red fox. These species may contribute to persistence of bovine tuberculosis in Doñana.

Bovine tuberculosis in a free ranging red fox (Vulpes vulpes) from Doñana National Park (Spain).

During 1997 and 1998, a survey of Iberian carnivores was conducted to study the epidemiology of bovine tuberculosis in the Doñana National Park and surrounding areas in southwestern Spain. Post-mortem examinations were done on seven red foxes (Vulpes vulpes), two Egyptian mongoose (Herpestes ichneumon), one weasel (Mustela nivalis), two genets (Genetta genetta), one Iberian lynx (Lynx pardinus), one Eurasian badger (Meles meles), and two polecats (Mustela putorius). Lesions suggestive of bovine tuberculosis were not detected but, in culture, Mycobacterium bovis was isolated from the retropharyngeal lymph nodes of one adult male red fox. This is the first report of M. bovis infection in red fox in Spain.

Genetic variability among Fasciola hepatica samples from different host species and geographical localities in Spain revealed by the novel SRAP marker.

A collection of483 samples representing Fasciola from six naturally infected host species and 16 localities in Spain, previously identified morphologically and genetically as Fasciola hepatica, was characterized by a novel genetic marker, namely sequence-related amplified polymorphism (SRAP), aiming to reveal genetic variability within F. hepatica in Spain. Visualization of amplification fragments was carried out on 6% denaturing polyacrylamide gels, followed by staining with 0.1% AgNO3 solution. Ten SRAP primer combinations were tested--six of them turned out to be polymorphic. Thirty-four representative F. hepatica samples from six host species and 16 geographical localities showed polymorphic banding patterns using SRAP primer combinations and were grouped into four major clusters using the unweighted pair-group method with arithmetic averages, indicating the existence of genetic variability within the examined F. hepatica samples. These four clusters were not related to particular host species and/or geographical origins of the samples. The results of the present study revealed that SRAP markers were useful in revealing sufficient polymorphism in F. hepatica samples from Spain and had implications for studying the population genetic structure of the Spanish F. hepatica. To our knowledge, this is the first application of SRAP marker to study genetic variation in parasites of human and animal health significance.