Using integrated wildlife monitoring to prevent future pandemics through one health approach.

In the One Health context, Integrated Wildlife Monitoring (IWM) merges wildlife health monitoring (WHM) and host community monitoring to early detect emerging infections, record changes in disease dynamics, and assess the impact of interventions in complex multi-host and multi-pathogen networks. This study reports the deployment and results obtained from a nationwide IWM pilot test in eleven sites representing the habitat diversity of mainland Spain. In each study site, camera-trap networks and sampling of indicator species for antibody and biomarker analysis were used to generate information. The results allowed identifying differences in biodiversity and host community characteristics among the study sites, with a range of 8 to 19 relevant host species per point. The Eurasian wild boar (Sus scrofa) was the most connected and central species of the host communities, becoming a key target indicator species for IWM. A negative relationship between biodiversity and disease risk was detected, with a lower number and prevalence of circulating pathogens in the sites with more species in the community and larger network size. However, this overall trend was modified by specific host-community and environmental factors, such as the relative index of wild boar - red deer interactions or the proximity to urban habitats, suggesting that human-driven imbalances may favour pathogen circulation. The effort of incorporating wildlife population monitoring into the currently applied WHM programs to achieve effective IWM was also evaluated, allowing to identify population monitoring as the most time-consuming component, which should be improved in the future. This first nationwide application of IWM allowed to detect drivers and hotspots for disease transmission risk among wildlife, domestic animals, and humans, as well as identifying key target indicator species for monitoring. Moreover, anthropogenic effects such as artificially high wildlife densities and urbanisation were identified as risk factors for disease prevalence and interspecific transmission.

Pestivirus in alpine wild ruminants and sympatric livestock from the Cantabrian Mountains, Spain.

Bovine viral diarrhoea virus (BVDV) and Border disease virus (BDV) were investigated at the wildlife-livestock interface in the distribution area of chamois in the Cantabrian Mountains, North-Western Spain. From 2010 to 2014, sera from sympatric wild (n=167) and domestic (n=272) ruminants were analysed for pestivirus antibodies by cELISA, virus neutralisation test (VNT) and for the presence of pestiviral RNA using a reverse transcription (RT)-PCR. Results showed a higher seroprevalence in cattle (59.4 per cent, 13/13 of herds) than in domestic small ruminants (5.9 per cent sheep, 2/8 of flocks; 0 per cent goats of 4 flocks) and wildlife (10.8 per cent in red deer, 0 per cent in roe deer and 0 per cent in Cantabrian chamois). High VNT titres were detected in two cattle herds, suggesting the circulation of BVDV-1 and BVDV-2 strains. BVDV-1 RNA was detected in one cattle calf by RT-PCR and sequencing. Conversely to other similar grazing systems, sheep flocks did not play a relevant role in the pestivirus epidemiology in this region. Pestivirus infections in wild ruminants were sporadic and most probably dependent on a domestic source.

Sex-related differences in body condition and serum biochemical parameters in red deer (Cervus elaphus) naturally infected with Mycobacterium bovis.

Although Mycobacterium bovis infection is commonly reported in red deer (Cervus elaphus), potential differences in the effects of infection on male and female animals in terms of body condition and clinical biochemistry have not been reported. Between November 2000 and January 2006, serum and biometrical data were collected post-mortem from 88 red deer. M. bovis-infected deer, particularly males, were typically older, heavier and in poorer body condition than uninfected animals. Serum triglyceride, cholesterol (both particularly in males) and total protein concentrations were lower, whereas serum creatinine (more evident in females), and immunoglobulin G and M concentrations were higher in the infected deer. These sex-related differences in the response to M. bovis infection in red deer should be considered when undertaking epidemiological assessments and designing disease control strategies as they may reflect differing roles of male and female animals as potential reservoirs or disseminators of disease.

Haematology and serum chemistry of Pyrenean chamois (Rupicapra pyrenaica) naturally infected with a border disease virus.

In 2005 and 2006 an outbreak of disease associated with border disease virus (BDV) infection caused high mortality in the Pyrenean chamois (Rupicapra pyrenaica) in the Catalan Pyrenees (NE Spain). The aim of this study was to determine values for different haematological and serum biochemical analytes in 32 free-ranging Pyrenean chamois affected by the disease and to compare them with those obtained from healthy chamois. In the affected chamois red blood cell counts, haemoglobin concentrations, packed cell volumes, mean corpuscular volumes and lymphocyte counts were all lower, while the neutrophil and platelet counts were higher. Glucose, lactate, triglycerides, creatinine, total protein concentrations and alkaline phosphatase activity were also lower, in contrast to the concentrations of total bilirubin, urea and aspartate aminotransferase activity, which were higher. Most of the observed changes could be associated with cachexia and inflammation in the affected chamois. Lymphopenia could be directly related to the BDV, which would lead to immunosuppression and explain the high rate of secondary infection observed in these animals.

Haloperidol and azaperone in drive-net captured Southern chamois (Rupicapra pyrenaica).

We investigated the effect of haloperidol and azaperone in drive-net captured Southern chamois (Rupicapra pyrenaica). Both tranquilizers have been successfully used in a wide range of wild species for reducing postcapture stress response. During 2005, 39 free-ranging chamois were captured, randomly injected intramuscularly with haloperidol (0.29 +/- 0.12 mg/kg; n=24), azaperone (1.1 +/- 0.82 mg/kg; n=6), or saline (0.5 ml; n=9), and restrained for 3 hr. Heart rate was higher in the treated chamois; erythrocyte parameters and total protein concentration decreased over time owing to splenic sequestration, hemodilution, vasodilation, and reflex tachycardia. Creatinine, sodium, and chloride remained stable only in the haloperidol-treated group, suggesting an improvement in renal perfusion. Nevertheless, the azaperone-treated chamois displayed higher body temperature, and both treated groups had higher serum muscular enzymes than the control group, suggesting higher muscle stress. These results lead us not to recommend the use of these tranquilizers-especially azaperone-as first-choice neuroleptics in chamois.

Serologic and virologic investigations into pestivirus infection in wild and domestic ruminants in the Pyrenees (NE Spain).

An outbreak of disease associated to a border disease virus was described in the Southern chamois (Rupicapra pyrenaica) in Spain in 2002. Sera and/or spleen samples from 57 mouflon, 15 red deer, 21 roe deer, 3 fallow deer, 55 sheep, 32 cattle, and 68 goats sharing the chamois habitat were studied. An antibody ELISA test yielded an inconclusive result in 2 mouflon and positive results in 5 goat sera. Comparative virus neutralization tests were performed on the 2 inconclusive mouflons, 3 of the 5 seropositive goats, 55 sheep and 32 cattle, using 6 pestivirus strains. Positive results were obtained in 1 mouflon, 2 goats, 69% of sheep and 78% of cattle. Virological investigations performed with an antigen ELISA test yielded negative results in 21 goats and 39 mouflons, the result in 1 mouflon being inconclusive. PCR performed on 12 goats and the inconclusive mouflon gave negative results. These results suggested that it is unlikely that chamois BDV is infecting wild and domestic ruminants.

High seroprevalence of Neospora caninum in the red fox (Vulpes vulpes) in the Pyrenees (NE Spain).

Antibodies to Neospora caninum were determined in the red fox (Vulpes vulpes) in the Pyrenees, north-eastern Spain. Sera from 53 red foxes (29 male and 24 female) were tested using a Neospora agglutination test (NAT). Seroprevalence at dilutions of 1/40, 1/80 and 1/160 was 69.8%, 47.2% and 7.5%, respectively. Sex differences were significant only at a dilution of 1:40. The high seroprevalence observed in red fox suggests that this species is highly exposed to N. caninum in this area.

Dermatophytosis caused by Trichophyton mentagrophytes in the southern chamois (Rupicapra pyrenaica) in the Eastern Pyrenees.

Two free-ranging southern chamois (Rupicapra pyrenaica) - one 4-year-old male and one 3-year-old male - were diagnosed with clinical dermatophytosis in the French and Spanish Eastern Pyrenees respectively. Skin samples were collected for microbiological studies, Trichophyton mentagrophytes being isolated and identified in both animals. The first chamois was found dead at the base of a cliff, and presented with alopecia and scaling on the dorsum and left forelimb. The second chamois showed grey-yellow, rough to raised scaling and crusting skin lesions at the base of the horns, around the eyes, dorsum of the nose, sternum, tail and limbs. Histological examination was carried out only on the second animal. The main lesions were orthokeratotic hyperkeratosis with focal parakeratosis, irregular to papillary epidermal hyperplasia, intracorneal neutrophilic pustules, perivascular to diffuse dermatitis with neutrophilic folliculitis and furunculosis. In those follicles with folliculitis and/or furunculosis, fungal hyphae and arthrospores associated with the follicular keratin and hair shafts were seen.

Effect of acepromazine on the signs of capture stress in captive and free-ranging roe deer (Capreolus capreolus).

The differences between the capture stress responses of captive and free-ranging roe deer (Capreolus capreolus) and their modulation by acepromazine, during a period of three hours' physical restraint after capture in drive-nets, were examined in 16 free-ranging and 16 captive roe deer. Eight of the free-ranging and eight of the captive animals received acepromazine intramuscularly, and the other eight free-ranging and eight captive deer received the same volume of saline. Heart rate, body temperature and haematological and serum biochemical parameters were analysed. In the groups treated with acepromazine, the heart rate stabilised sooner, and the red blood cell (RBC) count, haemoglobin concentration, packed-cell volume, the serum activities of creatine kinase (CK), aspartate aminotransferase (AST), alanine aminotransferase (ALT) and lactate dehydrogenase (LDH) and the concentrations of creatinine and lactate were significantly lower, and serum glucose started to decrease earlier, than in the untreated groups. Serum potassium levels decreased over time only in the untreated groups. The body temperature stabilised earlier, and the RBC count, haemoglobin concentration, serum CK, AST, ALT and LDH activities, and serum creatinine, lactate, cholesterol and glucose concentrations were significantly lower in the free-ranging roe deer than in the captive deer.

Haematological and serum biochemical values of southern chamois (Rupicapra pyrenaica).

Blood samples were taken from 75 free-ranging southern chamois (Rupicapra pyrenaica) captured in drive-nets in Catalonia, north-eastern Spain, and 20 haematological and 24 serum biochemical variables were analysed. The values were similar to those of other species of the Caprinae subfamily, except for cortisol, the concentration of which was higher. The red blood cell count (RBC), platelets and leucocytes, and the concentrations of cortisol, lactate, muscular enzymes and gamma-globulins were higher in summer than in spring, whereas the mean corpuscular volume, mean corpuscular haemoglobin, mean corpuscular haemoglobin concentration, and the concentrations of cholesterol, total bilirubin and creatinine were lower. Adult males had higher RBCs and haemoglobin values than females in summer, and lower leucocyte, lymphocyte and neutrophil counts than females and yearling males. The concentrations of triglycerides, total bilirubin, lactate, creatinine, urea, chloride and alpha2-globulins were higher in adult males than yearling males. In summer the adult females had higher values for platelets, lymphocytes, cortisol, sodium and muscular enzymes.