Detection of Mycoplasma conjunctivae in the eyes of healthy, free-ranging Alpine ibex: possible involvement of Alpine ibex as carriers for the main causing agent of infectious keratoconjunctivitis in wild Caprinae

Mycoplasma conjunctivae is considered the major cause of infectious keratoconjunctivitis (IKC) in Alpine ibex (Capra i. ibex) and chamois (Rupicapra r. rupicapra). While it is known that domestic sheep can act as healthy carriers for M. conjunctivae, this question has not been addressed in wild ungulates so far. In this study, bacteriological investigations and field observations were performed to assess whether free-ranging Alpine ibex can be healthy carriers of M. conjunctivae. Among 136 ibex without clinical signs of IKC, M. conjunctivae was identified 26 times (19.1%) by TaqMan PCR. To assess the potential pathogenicity of M. conjunctivae strains isolated from asymptomatic eyes, strains from three healthy ibex and from 15 IKC-ibex and IKC-chamois were analysed genetically by DNA sequence analysis of the variable part of the lppS gene. No significant differences were observed between strains from asymptomatic and clinically affected animals, reflecting the assumption that healthy ibex may act as carriers for M. conjunctivae strains that may be pathogenic for other individuals. Our results further indicate that development of IKC is associated with M. conjunctivae load in the eyes. In addition, a questionnaire survey revealed that IKC is generally less common in ibex than chamois and that infection in wild ungulates is not necessarily linked to the presence of sheep. These data support the hypothesis that apparently healthy ibex may be important in the epizootiology of IKC and indicate that host predilection may play a role in IKC development.     

Immobilising free-ranging Alpine chamois with xylazine, reversed with atipamezole

Between 1996 and 2005, 215 free-ranging Alpine chamois (Rupicapra rupicapra) were immobilised with xylazine hydrochloride. The 110 male and 105 female animals received a mean (sd) dose of 2.5 (0.6) mg/kg with a range from 1.4 to 4.8 mg/kg. The immobilisation was reversed in 201 of the animals with an intramuscular injection of 0.3 (0.1) mg/kg atipamezole (range 0.03 to 0.76 mg/kg), corresponding to a mean ratio of atipamezole:xylazine of 1:9.4 (4.3). All the chamois were immobilised, but shorter induction and recovery times, and deeper sedation with no reactions to handling were obtained in more than 80 per cent of the animals with doses of 2.6 to 3.6 mg/kg of xylazine, reversed with 0.26 to 0.36 mg/kg atipamezole (a ratio of 1:10), injected within 90 minutes.     

Severe outbreak of disease in the southern chamois (Rupicapra pyrenaica) associated with border disease virus infection

An outbreak of a previously unreported disease affecting southern chamois (Rupicapra pyrenaica) in the central Pyrenees (NE Spain) was recorded in 2001 and 2002. There was a marked temporal distribution, most animals being found between February and June. After the outbreak, the population was found to have decreased by about 42%, most probably due to the disease. We examined 20 affected chamois. Clinical manifestations included depression, weakness and movement difficulties in all cases. Three chamois presented abnormal behaviour, with absence of flight reaction, and 16 showed different degrees of alopecia with skin hyperpigmentation. At necropsy cachexia was observed in all animals, four chamois had abscesses in different parts of the body, four had pneumonia, one had an extensive subcutaneous infection on the head and neck and one had severe orchitis. Microscopic lesions were found in the brain, mainly edema, gliosis, espongiosis, cariorrexis and neuronal multifocal necrosis. A perivascular mononuclear inflammatory infiltrate was present in three of them. Skin lesions included marked follicular atrophy, mild to moderate epidermal hyperplasia with orthokeratotic hyperkeratosis and follicular hyperkeratosis, and hypermelanosis. In 13 chamois there were haemosiderin deposits in the spleen, and in three individuals kidney "cloissone" was observed. Intraeritrocitic parasites were detected either by direct observation or PCR in 8 of 17 chamois. A pestivirus was isolated and detected by RT-PCR from 12 of 13 affected chamois and antigenic characterized as border disease virus by monoclonal antibodies. This is the first time a border disease virus has been associated with an outbreak of a high-mortality disease in a wild species.     

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.     

Outbreaks of infectious keratoconjunctivitis in alpine chamois and ibex in Switzerland between 2001 and 2003

Five outbreaks of infectious keratoconjunctivitis (ikc) affecting alpine chamois and ibex in the western and central Swiss Alps were recorded in 2001 to 2003. Mycoplasma conjunctivae was identified from conjunctival swabs by means of a nested pcr in 27 of the 28 chamois tested. The outbreaks occurred in an area covering 1590 km(2). Deep valleys acted as a barrier to the spread of the disease. A total of 409 chamois and 33 ibex with clinical signs of ikc were reported. Most of the chamois were shot, primarily because they were blind or in poor general body condition. Almost a quarter were observed alive, and 16.9 per cent died as a result of ikc. Many of the affected animals were juveniles, and more females than males died of ikc. The disease was more common during the summer and autumn. The chamois affected by ikc were found at altitudes between 550 and 3200 m. The estimated overall mortality was less than 5 per cent, but more than 20 per cent may have died locally. Ibex affected by ikc were recorded in only two outbreaks. In six places, ibex with clinical signs of ikc were found before the first affected chamois appeared in the same area.     

Epidemiological study of border disease virus infection in Southern chamois (Rupicapra pyrenaica) after an outbreak of disease in the Pyrenees (NE Spain)

In 2001 and 2002, an outbreak of a previously unreported disease, associated with a border disease virus (BDV), caused high mortality in the Southern chamois (Rupicapra pyrenaica) population in the Alt Pallars-Aran National Hunting Reserve in the Catalan Pyrenees (NE Spain). Between 2002 and 2006, sera and/or tissue samples taken from 116 healthy chamois shot during the hunting season, plus 42 from chamois affected by different diseases, were studied. A blocking enzyme-immunosorbent assay (ELISA) was used to study pestivirus seroprevalence in 114 healthy hunted and 31 diseased chamois, yielding positive results in 73.7 and 22.6% of the chamois, respectively. Comparative virus neutralization tests (VNT) performed on 42 seropositive samples with 6 pestivirus strains yielded statistically higher titres to BDV Spain 97, followed by BDV chamois, BDV 137/4, BDV Moredun, Bovine Diarrhoea virus-1 (BVDV-1) NADL and BVDV-2 atypical. Virological investigations for pestivirus detection were performed using an antigen ELISA test in 82 healthy and 18 diseased chamois, RT-PCR in 16 healthy and in all diseased chamois, and virus isolation in 14 diseased chamois. No viral antigen was detected in any of the healthy animals. A pestivirus, characterized as BDV by monoclonal antibodies, was detected in the 10 chamois showing clinical signs consistent with BDV infection. Sequence analysis in the 5' untranslated region (5'-UTR) revealed that they were grouped into the BDV-4 genotype. In the remaining chamois, infectious keratoconjunctivitis, pneumonia, trauma and contagious ecthyma were diagnosed. The cause of death was unknown in five chamois. The results suggest that the infection has become endemic in the population and that it could have a significant impact on chamois population dynamics.     

Effects of sarcoptic mange on serum proteins and immunoglobulin G levels in chamois (Rupicapra pyrenaica) and Spanish ibex (Capra pyrenaica)

Three groups of chamois (Rupicapra pyrenaica) and three groups of Spanish ibex (Capra pyrenaica) were established to study the effects of sarcoptic mange on serum proteins and immunoglobulin G (IgG) levels. The first group of chamois consisted of 22 healthy Pyrenean chamois (R. pyrenaica pyrenaica) from a non-infested area, the second group consisted of 20 healthy Cantabrian chamois (R. p. parva) from an area where sarcoptic mange has been reported since 1994 and the third group consisted of 16 Cantabrian chamois from the same area but naturally infested by Sarcoptes scabiei. The first group of Spanish ibex was 39 healthy animals from a sarcoptic mange non-infested area, the second group was 23 healthy animals from a sarcoptic mange infested area and the third group consisted of 20 animals from the same area but naturally infested with the parasite. Blood samples were taken after killing the animals as part of hunting programmes. Values for total proteins, gamma-globulin and IgG were higher in infested and healthy chamois from the infested area compared to healthy chamois from the non-infested area, and IgG levels were higher in infested chamois compared to healthy-exposed chamois. Values for alpha2-globulin were higher in healthy Cantabrian chamois. In Spanish ibex, albumin, alpha2-globulin and IgG levels were lower in the healthy Spanish ibex from the non-infested area than in healthy animals from an infested area. The differences found in the chamois were indicative of the establishment of a humoral antibody response in the animals in contact with the disease. As the IgG levels were not significantly different between healthy and infested Spanish ibex from the same area, a different pattern of chronic infection with humoral response to the disease was suggested.     

Retrospective study of pestivirus infection in Pyrenean chamois (Rupicapra pyrenaica) and other ungulates in the Pyrenees (NE Spain)

In 2001 a new Pestivirus (Family Flaviviridae) was associated with an outbreak of a previously unreported disease in Pyrenean chamois (Rupicapra pyrenaica) in the Pyrenees (NE Spain). Molecular characterization assigned this virus to the Border Disease Virus (BDV) cluster, BDV-4 genotype. A retrospective study was performed in archived sera and spleen of 74 Pyrenean chamois and in archived sera of 28 mouflon (Ovis ammon), 56 red deer (Cervus elaphus), 43 roe deer (Capreolus capreolus) and 29 fallow deer (Dama dama) from the Pyrenees between the years 1990 and 2000. Thirty six of 74 (48.6%) sera of Pyrenean chamois, one of mouflon and one of red deer were positive by an ELISA antibody test. Comparative virus neutralization tests were performed on 26 seropositive chamois, one mouflon and one red deer, using five pestivirus strains. An ELISA antigen test was performed on 37 seronegative chamois and yielded positive results in one chamois and inconclusive result in two. RT-PCR and virus isolation performed on spleen samples from these three animals gave positive results in the positive and one inconclusive animal. Sequence analysis in the 5' unstranslated region revealed that they were grouped into the BDV-4 genotype. Virological and serological data of the present study indicate that BDV infection has been present in the chamois population since at least 1990, 11 years before the first outbreak of disease. Therefore, the emergence of the disease in 2001 is apparently due to other factors rather than the introduction of a new virus in the chamois population.     

Lung-tissue extract as an alternative to serum for surveillance for brucellosis in chamois

Serological surveys are the most-used way to study diseases in free-ranging wild animals. However, the difficulty in obtaining a sufficient number of suitable serum samples is a major problem. To resolve this problem, we investigated the possibility of using lung-tissue extract in place of blood serum for searching for antibodies against Brucella abortus. Antibodies titres against B. abortus was tested from blood serum and lung-tissue extract from 112 chamois and 99 cattle. Although in complement-fixation-test, lung-tissue extract titres usually were one-to three-fold lower than serum titres, there was a good agreement between serum and lung-tissue extract positivity both in chamois and in cattle. The lung-tissue extract appears a suitable resource in monitoring brucellosis in chamois.     

Sarcoptic mange in red deer from Spain: improved surveillance or disease emergence?

Concern about emerging diseases has risen in recent years, and multihost situations have become increasingly relevant for wildlife management and conservation. We present data on Asturias, northern Spain, where 80 mangy red deer (Cervus elaphus) have been found since the beginning of the epizootic in chamois (Rupicapra pyrenaica parva) in 1993. We combine field and necropsy data with the results of a serosurvey using an in-house ELISA test to evaluate if deer mange due to Sarcoptes scabiei is an emerging disease in this area. The mean number of deer mange cases per year was 5, with a maximum of 16. No significant relationship was detected between monthly temperatures, rainfall or number of days with snow cover and the annual number of sarcoptic mange cases in red deer. Only 4 mangy red deer (5%) were detected outside the limits of scabietic chamois distribution during the same year, and all were less than 2500 m away from that limit. The longest distance reported between two consecutive mangy deer locations was 18 km. Mange cases were significantly more frequent in stags than in hinds and in adults than in juvenile deer. The time of the first mange detection in chamois in each sector, year with minimum number of chamois recorded, year with maximum chamois population decline rate and chamois density offered no significant correlation with red deer mange cases appearance moment and frequency. In the mange affected area, ELISA testing of 327 blood samples from hunter-harvested deer without obvious mange-compatible lesions revealed only 4 seropositive animals. All 83 sera from hunting preserves without clinical cases yielded negative ELISA results. According to these epidemiological data mange does not seem to threaten red deer populations in Asturias. However, continued monitoring of deer health and ELISA testing for sarcoptic mange is advisable.     

Seroprevalence of pestivirus in four species of alpine wild ungulates in the High Valley of Susa, Italy

Wildlife, once infected, can serve as a reservoir of infectious diseases that form a constant threat to domestic livestock. To make control and eradication programs successful in the long-term, presence of pestivirus in wildlife populations should be monitored. The goal of this study was to investigate seroprevalence of pestivirus in four alpine wild ungulates in the High Valley of Susa, north-west Italy. Species studied were: red deer (Cervus elaphus), roe deer (Capreolus capreolus), wild boar (Sus scrofa) and chamois (Rupicapra rupicapra). A further goal was using virus neutralisation tests (VNT) for four strains of pestivirus in chamois and wild boar. Three hundred and seventy-five serum samples collected during the hunting season of 1999 were tested for pestivirus specific antibodies. Positive sera of chamois and wild boar were subsequently tested in a VNT with four major subtypes of pestivirus, and virus isolation was performed. No antibodies were found in the 73 samples of roe deer, while 7 (12.5%), 8 (5.9%) and 28 (25.5%) of 56, 136 and 110 samples of wild boar, red deer and chamois were ELISA-positive, respectively. Different ranges of titers were found in the VNT and no pestivirus was isolated in the ELISA-positive wild boar and chamois samples. Several possibilities, which might explain the high seroprevalence in chamois are discussed. Pestivirus antibodies were found in three out of four large alpine ungulates in the High Valley of Susa. Seroprevalence was particularly high in chamois. Further investigation is needed to characterise the pestiviruses that circulate in these animals.     

Experimental infection of chamois (Rupicapra pyrenaica parva) with Sarcoptes scabiei derived from naturally infected goats

Two chamois (Rupicapra pyrenaica parva) out of a group of three were experimentally infected with Sarcoptes scabiei derived from a naturally infected domestic goat. One of the chamois presented the first clinical manifestations (papules and desquamation) at 7 days post-infection, after 22 days crusts and alopecia appeared and after 41 days pruritus. The other chamois presented desquamation after 15 days and papules after 21 days and crusts and alopecia after 31 days. All these clinical manifestations continued to spread and when the animals were treated at 84 days post-infection, pruritus, papules and crusts were first to disappear, there being no evidence of their presence at 99 days post-infection, when the second treatment dose was applied. The desquamation and alopecia disappeared at 114 days post-infection, by which stage both animals were considered to have been cured. The results of the skin scraping was negative in both chamois until 54 days post-infection and it became negative again after 84 days, when the first treatment dose was applied. Biopsies showed different levels of hyperkeratosis and a marked epidermic hyperplasia with formation of small crusts. Superficial epidermis presented marked vasodilatation and also infiltrated inflammation. None of the biopsies carried out showed the presence of parasites. The non-infected chamois, which was kept in the same compound as the other two, did not present any clinical manifestations compatible with infection by S. scabiei throughout the entire period of the experiment.     

Detection of space-time clusters and epidemiological examinations of scabies in chamois

In this paper some methodological approaches towards the detection of temporal, spatial and space-time clusters are presented and discussed in context with epidemiological factors of scabies in chamois. This presentation comprises in particular the use of the Scan Test (Wallenstein, 1980) for the detection of temporal clusters and the tests developed by Knox (1964) and Mantel (1967) for the testing of spatiotemporal interactions. Between 1952 and 1998 a total of 1689 cases of scabies in chamois and six cases in ibex were documented in 137 game preserves of the Austrian province of Styria. The evaluations indicate the presence of spatial and temporal clusters. Some possibilities for the spread of scabies in chamois are presented and reveal ibex as a probable vector for scabies transmission between chamois populations. The information may lead to strategies for controlling the spread of the disease.     

Serological study of a population of alpine chamois (Rupicapra r rupicapra) affected by an outbreak of respiratory disease

A serological survey of respiratory virus infections was carried out from 1998 to 2001 in Lecco province, Italy, as part of a health monitoring programme in a population of alpine chamois, many of which died of pneumonia in autumn and winter 2000 to 2001; 194 carcases of all age classes were found over a short period and in a small area. Eighteen of them, which were examined postmortem, consistently showed signs of severe fibrinous lobar pneumonia or catarrhal bronchopneumonia. Samples of serum from 145 chamois collected from hunted animals and carcases were tested by a virus neutralisation test against bovine respiratory syncytial virus (BRSV), bovine viral diarrhoea virus, bovine herpesvirus type 1 and parainfluenzavirus type 3. Positive results were detected only for BRSV. The area was divided into two subunits on the basis of the distribution of deaths; in the areas where fatalities were observed there was a significant increase of BRSV titres at the beginning of the outbreak. Furthermore, during the 2000 and 2001 hunting seasons antibody titres to BRSV were significantly higher in the areas where mortality occurred. The roe deer living in the same area were not affected by pneumonia and had a low prevalence of titres to BRSV which did not vary during the period of the study.     

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.     

Astrocytoma in a chamois

Astrocytomas represent the most common cerebral tumors in humans and in animals, and the fibrillary cytological subtype is the most frequently observed. In this report and for the first time, a thalamic astrocytoma is described in a chamois showing depressed mentation, pleurothotonus and circling to the right side.     

Babesia divergens-like organisms from free-ranging chamois (Rupicapra r. rupicapra) and roe deer (Capreolus c. capreolus) are distinct from B. divergens of cattle origin - an epidemiological and molecular genetic investigation

In 2005 and 2006, three adult female chamois (Rupicapra r. rupicapra) were found dead with signs of acute babesial infection in the eastern Swiss Alps. PCR on DNA extracted from blood or spleen of the carcasses revealed sequence identity of the amplified part of the 18S rRNA gene with GenBank entries attributed to Babesia divergens of cattle origin or B. capreoli of wild ruminant origin which have never been described before in this region. Examination of 424 blood samples from 314 head of cattle from this area by IFAT, microscopy and PCR provided no evidence for babesial infection. Six of 887 ticks collected from cattle were PCR-positive, and sequencing revealed Babesia sp. genotype EU1 in five and B. divergens/B. capreoli in one of them. A Babesia isolate of chamois, two isolates of roe deer from the same region and one isolate of a roe deer from the north-western Swiss Alps were genetically compared with two Swiss B. divergens isolates of cattle origin by analysing the genomic rDNA locus. Whereas the near full length sequences of the 18S rRNA gene were virtually identical among all six isolates (>99.4% identity), distinct differences between the two isolates from cattle on the one hand and the four isolates from free-ranging ruminants on the other hand were observed in the sequences of the internal transcribed spacers 1 and 2 (ITS1, ITS2) and part of the 28S rRNA gene. These results indicate that, albeit genetically very closely related, these babesial organisms from cattle and from free-ranging ruminants indeed are distinguishable organisms with different host specificities, and they support the use of the discrete species name B. capreoli for the B. divergens-like organisms from chamois and roe deer.     

Mycobacterium avium subsp. paratuberculosis in wild animal species and cattle in Styria/Austria

Infections with Mycobacterium ovium ssp. paratuberculosis (M. paratuberculosis) are increasingly recognised worldwide. In addition to an increased prevalence of paratuberculosis in Austrian cattle herds, recent years have also shown a rise in infections with M. paratuberculosis in wild red and roe deer, chamois and mouflon. During the period from June 2002 to September 2004, mesenteric lymph nodes were taken from a total of 483 wild animals hunted or found dead and from 338 deceased cattle. Samples were analysed using PCR and cultivation methods. In the case of pathomorphological changes or anamnestic indications, investigations also included an analysis of organ samples (e.g. liver, lung) or foetuses. The tests revealed that 129 wild animal samples (red deer, roe deer, chamois, mouflon, fallow deer, ibex, foxes, mountain hare, yellow-necked field mouse, and capercaillie) contained M. paratuberculosis. The major symptoms in the wild aninodes. Evidence of diarrhoea was only observed in about 15% of the positive cases. The study for the first time provided evidence of intrauterine transmission of M. paratuberculosis in red deer (3 cases) and chamois (1 case) and succeeded in the isolation of the pathogen from the liver, lung and subcutaneous granulomas of wild animals. Of the total of 338 mesenteric lymphnodes of cattle from 303 herds, 80 samples from 77 herds tested positive for paratuberculosis. Twenty-two wild animal and 3 cattle isolates have so far been molecularly typed using IS900-RFLP and RAPD analyses in order to prove epidemiological relationships between occurrences in cattle and wild animals. The increase of paratuberculosis in wild animal species is assumed to have been caused by the purchase of animals, a strong increase in suckler cow farming (cow-calf herds) with a concentration of pathogens in the environment and by inadequate feed hygiene for wild animals.     

Mesenchymal tumor in a female chamois. A case report

A free-living 15-year old female chamois was shot because of illness and emaciation. The examination of the internal organs revealed numerous firm whitish tumour nodules in liver,spleen, mesenterium and peritoneum. Histologically the nature of tumour cells was identified as a metastasizing malignoma of mesenchymal origin. In addition to the blastoma, the parasite burden of the gastrointestinal tract and lungs did very likely contribute to the bad bodily condition of the chamois.     

Biotin-avidin amplified ELISA for detection of antibodies to Sarcoptes scabiei in chamois (Rupicapra spp.)

Scabies is a major threat to the well being of mountain-dwelling Bovid hosts, Rupicapra rupicapra and Rupicapra pyrenaica. Severe outbreaks are in progress over a significant part of their distribution area and resource managers demand improved methods to monitor, analyse and possibly forecast the spread and effects of scabies at the population level. An amplified capture enzyme-linked immunosorbent assay was developed to detect antibodies to Sarcoptes scabiei in chamois (Rupicapra spp.) serum. The method used the biotin-avidin amplification system and was validated on a panel of 144 serum samples, of which 40 were obtained from scabietic and 104 from healthy unexposed individuals originating from a scabies-free area. The antigen, a whole body extract of the various developmental stages of S. scabiei, was prepared from mites actively leaving the skin lesions of naturally infested red foxes (Vulpes vulpes). The resulting LAB-ELISA was characterised by 93% sensitivity, 97% specificity and a high degree of repeatibility. A single seroreactor was found amongst 32 chamois affected with skin pathologies other than scabies, including infestations by other Acarina (Trombicula spp. and Ixodid ticks). Antibodies to S. scabiei were present in 26 out of 169 sera (15.4%) obtained by clinically healthy chamois within a scabies outbreak area, indicating that asymptomatic infestations by S. scabiei can be revealed by serological methods in the studied Caprinae hosts.