Comparative evaluation of effort, capture and handling effects of drive nets to capture roe deer (Capreolus capreolus), Southern chamois (Rupicapra pyrenaica) and Spanish ibex (Capra pyrenaica)

The objective of this study is to assess the usefulness of drive nets to capture roe deer (Capreolus capreolus), Southern chamois (Rupicapra pyrenaica) and Spanish ibex (Capra pyrenaica), comparing the results obtained with other capture methods and amongst the three species. Sixty-five drive net capture operations using beaters were conducted from January 1998 to September 2004. A total of 161 wild ungulates (31 roe deer, 95 Southern chamois and 35 Spanish ibexes) were captured. The average number of animals captured per operation was 1.07 for roe deer, 3.96 for Southern chamois and 2.92 for Spanish ibex. The average number of person–days per captured animal was 21.5, 7.1 and 10.6 for roe deer, Southern chamois and Spanish ibex, respectively. Specificity was 100% for Southern chamois and Spanish ibex (only the target species captured) and 77.5% for roe deer. Risk for the animals (mortality plus injuries) was 3.23% for roe deer, 5.27% for Southern chamois and 0% for Spanish ibex, whereas injuries to the operators occurred with 3.1% of the handled animals. Sex ratio was skewed towards females in roe deer, towards males in Southern chamois and balanced in Southern chamois. Drive nets showed good performance, although many operators were required. Safety for the animals and specificity were higher than traditionally attributed to this capture method. It is concluded that drive nets are an efficient and safe method to capture many ungulate species.     

Cohabitation of a <Emphasis Type="Italic">Brucella melitensis</Emphasis> infected Alpine ibex (<Emphasis Type="Italic">Capra ibex</Emphasis>) with domestic small ruminants in an enclosure in Gran Paradiso National Park,in Western Italian Alps

After the first report of Brucella melitensis infection from a 7-year-old alpine ibex (Capra ibex) buck living in Gran Paradiso National Park (GPNP), further studies demonstrated the presence of the infection in ibex and chamois. Considering that livestock herds keep on sharing pastures with more than 3,500 ibex and 9,000 chamois in the park, our aim was to demonstrate under controlled conditions the possibility of Brucella infection passing from wild ruminants to livestock. A 7-year-old male alpine ibex with clinical signs of brucellosis and serologically positive was released in a 5,000 m² enclosure together with five goats and two sheep rams. Due to poor condition, ibex was suppressed at day 40, domestic ruminants stayed into the enclosure potentially contaminated by ibex for further 38 days. During this period, we had monitored our animals taking blood from domestic ruminants every 15 days and tested the serum to Rose Bengal agglutination test and Complement Fixation test. Domestic animals tested negative at serology at all sampling time and at isolation, while B. melitensis biovar 3 was isolated from ibex tissues. Our data show that transmission of infection from ibex to livestock is not easy. After 40 days of strict cohabitation and 38 days of permanence in an area where an infected ibex lives, no one of the domestic animals contracted infection. In spite of the limitation of our field trial, we have demonstrated that long direct and indirect contact between alpine ibex and domestic animals will not easily lead to an infection of the latter. Further investigations are needed to confirm our results and evaluate the effective risk of B. melitensis transmission from alpine ibex to livestock.     

Techniques for noninvasive genetic monitoring of Alpine Ibex <Emphasis Type="Italic">Capra ibex</Emphasis>

Noninvasive sampling is of increasing importance for the molecular genetic monitoring of wild animal populations, although reduced quality and quantity of such samples’ DNA can affect genetic data and their subsequent interpretation. Consequently, we performed a pilot study to establish a feasible approach for the genetic investigation of free-ranging Alpine ibex Capra ibex Linnaeus, 1758 populations. Establishing an ibex-specific PCR-RFLP based on Cytochrome b gene differences allowed the discrimination of noninvasive ibex samples from those of other sympatric ungulates. In addition, we established a quantitative PCR for ibex samples. The quantification of 35 faecal samples clearly exhibited a strong variability of DNA contents among samples and individuals. Furthermore, we performed threefold genotyping experiments on six microsatellite loci to determine the extent of genotyping errors in reference to blood samples of the respective individuals. The analyses exhibited a strong dependence of erroneous microsatellite genotypes on the starting amount of template DNA. Variability in reliability was observed between individual loci, resulting in a mandatory high DNA concentration necessary for consistent genotyping. This study serves as basis for further ibex research and we propose the application of DNA quantification of faecal samples to focus genotyping efforts solely on suitable samples.     

Nucleolus-organizer regions and heterochromatin in three species of Bovidae

Ag-NOR staining and a counterstain-enhanced fluorescence technique (chromomycin A₃/distamycin A/DAPI staining = CDD-method) have been applied to ibex (Capra ibex L.), chamois (Rupicapra rupicapra L.) and bison (Bison bison L.) chromosomes.Chromomycin A₃ visualization led to a well defined R-banding pattern along the chromosome arms and to a clear demonstration of centric heterochromatic bands of variable size. The nucleolus organizer regions (NORs) were found in the telomeric regions of the chromosomes 2, 3, 4, 5 and 28 of the ibex, of the chromosomes 1/3 (short arm), 2, 4, 5 and 28 of the chamois and of the chromosomes 2, 3, 4, 11 and 28 of the bison.     

Is the development of infectious keratoconjunctivitis in Alpine ibex and Alpine chamois influenced by topographic features?

Infectious keratoconjunctivitis (IKC) caused by Mycoplasma conjunctivae is a widespread ocular affection of free-ranging Caprinae in the Alpine arc. Along with host and pathogen characteristics, it has been hypothesized that environmental factors such as UV light are involved in the onset and course of the disease. This study aimed at evaluating the role of topographic features as predisposing or aggravating factors for IKC in Alpine chamois (Rupicapra rupicapra rupicapra) and Alpine ibex (Capra ibex ibex). Geospatial analysis was performed to assess the effect of aspect (northness) and elevation on the severity of the disease as well as on the mycoplasmal load in the eyes of affected animals, using data from 723 ibex and chamois (583 healthy animals, 105 IKC-affected animals, and 35 asymptomatic carriers of M. conjunctivae), all sampled in the Swiss Alps between 2008 and 2010. An influence of northness was not found, except that ibex with moderate and severe signs of IKC seem to prefer more north-oriented slopes than individuals without corneal lesions, possibly hinting at a sunlight sensitivity consequent to the disease. In contrast, results suggest that elevation influences the disease course in both ibex and chamois, which could be due to altitude-associated environmental conditions such as UV radiation, cold, and dryness. The results of this study support the hypothesis that environmental factors may play a role in the pathogenesis of IKC.     

Cross‐amplification of 10 new isolated polymorphic microsatellite loci for red mullet (Mullus barbatus) in striped red mullet (Mullus surmuletus)

Ten polymorphic dinucleotide microsatellite loci were isolated and characterized for the red mullet (Mullus barbatus). Allele variability was tested on both the red mullet and its congener the striped red mullet (Mullus surmuletus). Characterization of 30 individuals of both species from the western Mediterranean showed moderate to high allelic diversity ranging from two to 26 alleles per locus (mean 10.9). Three loci showed departures from Hardy–Weinberg proportions. No evidence of significant association between genotypes at pairs of loci was observed. These polymorphic loci could be suitable for population genetic assessments of both species.     

Descriptive epidemiology of a scabies epidemic in chamois in the Dolomite Alps,Italy

In 1995, the hitherto Sarcoptes-free alpine chamois (Rupicapra rupicapra) of the Dolomite Alps (Italy) were affected by scabies for the first time after the spread of the disease from a neighbouring focus in Austria. Since then, four Agencies cooperated to warrant monitoring of the spatial and temporal progress of the outbreak and further active surveillance was carried out within an intensive study area. In ten years, 15 meta-populations numbering 10,000 chamois and 210,000 ha were encompassed with a maximum distance of 55 km from the index case. “Oil spot” advancement of the epidemic was observed together with “jumps” of 9 to 20 km followed by spreading of the infection to the intervening population units. Demographic decline of the four meta-populations so far affected for a minimum of six years ranged between 49 and 77% (), whereas similar post-epidemic densities of 1.1 to 1.7 heads/100 ha () were recorded. Contiguous resistant and sensitive herds with similar pre-epidemic density were detected on a smaller population scale, suggesting complementary mechanisms intervening besides density-dependence as determinants of the outbreak outcome. No sex or age class showed a higher sensitivity to scabies out of a sample of 1,696 infected chamois. A peak prevalence of free-ranging chamois bearing overt scabietic lesions was observed in January and February. Cases were only sporadically diagnosed in other sympatrc wild ruminants (Cervus elaphus, Capreolus capreolus and Ovis gmelini musimon), whereas all four alpine ibex (Capra ibex ibex) herds living in the surveyed area suffered from scabies-induced decline.     

Microsatellite markers isolated from polyploid wood‐sorrel Oxalis alpina (Oxalidaceae)

Twelve polymorphic microsatellite loci were isolated from polyploid alpine wood‐sorrel, Oxalis alpina (Oxalidaceae), and optimized for future studies of its breeding system. The loci were screened for variability among 72 individuals from Pinos Altos, New Mexico. The primers amplified loci with allele number ranging from two to 17 per locus and with estimates of Nei's genetic diversity varying from 0.10 to 0.99. These primers provide an opportunity to use polymorphic DNA markers to study the causes of breeding system variability in this species.     

Occurrence, quantification, and genotyping of Mycoplasma conjunctivae in wild Caprinae with and without infectious keratoconjunctivitis

Mycoplasma conjunctivae, the causative agent of infectious keratoconjunctivitis (IKC), was recently detected in asymptomatic Alpine ibex (Capra ibex ibex). This suggested that an external source of infection may not be required for an IKC outbreak in wildlife but might be initiated by healthy carriers, which contradicted previous serologic investigations in chamois. Our aims were to 1) assess the prevalence of M. conjunctivae among asymptomatic ibex and Alpine chamois (Rupicapra rupicapra rupicapra) and its frequency in IKC-affected animals, 2) determine mycoplasma loads in different disease stages, and 3) characterize the M. conjunctivae strains involved. Eye swabs from 654 asymptomatic and 204 symptomatic animals were collected in diverse Swiss regions between 2008 and 2010, and tested by TaqMan real-time PCR. Data analysis was performed considering various patterns of IKC occurrence in the respective sampling regions. Strains from 24 animals were compared by cluster analysis. Prevalence of M. conjunctivae was 5.6% (95% confidence interval [CI]: 3.7-8.1%) in asymptomatic ibex and 5.8% (CI: 3.0-9.9%) in asymptomatic chamois, with significant differences between years and regions in both species. Detection frequency in symptomatic animals was significantly higher during IKC outbreaks than in nonepidemic situations (i.e., regular but low incidence or sporadic occurrence). Mycoplasma load was significantly lower in eyes from healthy carriers and animals with mild signs than from animals with moderate and severe signs. Although some strains were found in both asymptomatic and diseased animals of the same species, others apparently differed in their pathogenic potential depending on the infected species. Overall, we found a widespread occurrence of M. conjunctivae in wild Caprinae with and without IKC signs. Our results confirm the central role of M. conjunctivae in outbreaks but suggest that other infectious agents may be involved in IKC cases in nonepidemic situations. Additionally, presence and severity of signs are related to the quantity of M. conjunctivae in the eyes rather than to the strain. We propose that individual or environmental factors influence the clinical expression of the disease and that persistence of M. conjunctivae in populations of wild Caprinae cannot be excluded.     

Genetic variability analysis of Giant river cattish (<Emphasis Type="Italic">Sperata seenghala</Emphasis>) populations from Indus river system by RAPD-PCR

The Giant river catfish, Sperata seenghala (Sykes) is commercially very important fish species of South Asia. Genetic variability between its populations collected from two rivers i.e. river Sutlej and river Beas of Indus river system in India were examined using randomly amplified polymorphic DNA analysis. Total 38 fish samples were collected from river Sutlej whereas 46 fish samples were collected from river Beas. Total 40 primers were screened, out of these 7 were selected for studying polymorphism which produced a total of 64 RAPD loci in two populations. Percentage polymorphic loci calculated following 95% criterion was 89.06 % for Beas population as compared to 95.31 % for Sutlej population. Moderate level of genetic divergence (genetic distance of 0.0486) between both the populations suggests distinct population substructure of giant river catfish in both the rivers.     

Genetic variation and phylogenetic relationship between two species of yellow catfish, <Emphasis Type="Italic">Horabagrus brachysoma</Emphasis> and <Emphasis Type="Italic">H. nigricollaris</Emphasis> (Teleostei: Horabagridae) based on RAPD and microsatellite markers

The two species of yellow catfish, Horabagrus brachysoma and H. nigricollaris are categorized as ‘endangered’ and ‘critically endangered’ respectively in their wild habitat. Proper knowledge of genetic structure and variability of these endangered species are highly essential for the management, conservation and improvement of fish stocks. Therefore, genetic variation and phylogenetic relationships between these species of yellow catfish sampled from Chalakkudy River in the hot spot of biodiversity-Western Ghats region, Kerala, India were analyzed by using Random amplified polymorphic DNA (RAPD) and microsatellite markers. 85 RAPD and five microsatellites loci were detected to analyze the genetic variation and phylogenetic relationships among these species. Out of 85 RAPD loci produced only 52.94% were polymorphic whereas in microsatellite, all 5 loci were polymorphic (100%). Species-specific RAPD bands were found in both species studied. In microsatellite, the number of alleles across the five loci ranged from 1 to 8. The observed heterozygosities in H. brachysoma and H. nigricollaris were 0.463 and 0.443, respectively. Here, both RAPD and microsatellite methods reported a low degree of gene diversity and lack of genetic heterogeneity in both species of Horabagrus which strongly emphasize the need of fishery management, conservation and rehabilitation of these species.     

A set of 21 polymorphic microsatellites in Alpine chamois (Rupicapra rupicapra)

A cross‐species amplification test of 39 bovid microsatellite markers was carried out on chamois (Rupicapra rupicapra), a species with a high economic and tourist value, for which a number of life‐history traits remain to be quantified. Of 39 primer pairs tested, 26 gave a specific amplification product and 21 were polymorphic. Locus‐specific allelic richness ranged from two to six. Our results suggest that these loci will be useful for investigating basic population genetics as well as life‐history traits, such as dispersal.     

Integrating phylogeographic patterns of microsatellite and mtDNA divergence to infer the evolutionary history of chamois (genus <Emphasis Type="Italic">Rupicapra</Emphasis>)

Background  

The chamois, distributed over most of the medium to high altitude mountain ranges of southern Eurasia, provides an excellent model for exploring the effects of historical and evolutionary events on diversification. Populations have been grouped into two species, Rupicapra pyrenaica from southwestern Europe and R. rupicapra from eastern Europe. However, a previous study of cytochrome b revealed that the two proposed species were non-monophyletic. The reconstruction of phylogenetic relationships between animal species often depends on the markers studied. To further elucidate the evolutionary history of chamois, we extended earlier studies by analysing DNA sequences of four mitochondrial regions (ND1, 12S, tRNApro and Control Region) and microsatellites (20 loci) to include all subspecies and cover its entire distribution range.     

Isolation and characterization of ten novel microsatellite loci in the red-winged tinamou (<Emphasis Type="Italic">Rhynchotus rufescens</Emphasis>, Tinamiformes,Aves) and cross-amplification in other tinamous

We describe the isolation and characterization of ten microsatellite loci from the red-winged tinamou (Rhynchotus rufescens) and also evaluated the cross-amplification of these loci and other ten loci previously developed for the great tinamou (Tinamus major) in other tinamous. Genetic variability was assessed using 24 individuals. Six loci were polymorphic with moderate to high number of alleles per locus (2–12 alleles) and showed expected heterozygosity (HE) ranging from 0.267 to 0.860. All loci conformed to the Hardy–Weinberg expectation and linkage disequilibrium was not significant for any pair of loci. This battery of polymorphic loci showed high paternity exclusion probability (0.986) and low genetic identity probability (4.95 × 10⁻⁵), proving to be helpful for parentage tests and population analyses in the red-winged tinamou. The cross-amplification was moderate where of the 160 locus/taxon combinations, 46 (28.75%) successfully amplified.     

Development and characterization of novel tetra‐ and dinucleotide microsatellite markers for the French Polynesia black‐lipped pearl oyster,Pinctada margaritifera

Ten microsatellite loci were developed for the black‐lipped pearl oyster Pinctada margaritifera with a magnetic bead enrichment protocol. These tetra‐ and dinucleotide markers were polymorphic, with 10 to 43 alleles observed in 97 individuals from two Tuamotu atoll populations. Most loci revealed significant genic differentiation between the two populations and also exhibited some degree of heterozygote deficiencies, probably due to the presence of null alleles. These loci should be very useful to describe genetic structure, genetic variability and reproductive success in the various aquaculture and wild populations of pearl oyster in French Polynesia.     

Genetic divergence between populations of two closely related troglobitic beetle species (Speonomus: Bathysciinae,Coleoptera)

Genetic variability and divergence at 13 enzyme loci were studied in two species of troglobitic beetles (Speonomus zophosinus andhydrophilus 12 populations) collected from the Pyrenees mountains. These allopatric species exhibit a high degree of specialization to underground life and a very small geographical range. Four diagnostic loci arePgi, Est-2, Pac-1, Phi. Within each species polymorphic loci exhibited marked spatial variation of allele frequencies, sometimes over short distances (5–10 km). The levels of genetic variability are comparable to those observed in non-cave invertebrates (frequency of polymorphic loci: 0.31; average expected heterozygosity: 0.112±0.015). Moreover, a single species showed variability in the average level of heterozygosity (0.06 to 0.172). Patterns of genetic differentiation among species were also studied; mean genetic distance (D: 0.263±0.010) between the twoSpeonomus species was of the same order of magnitude as most data reported in the literature between species. A significant heterozygote deficiency occurs in local populations and for all loci. This deficit does not vary from locus to locus. Several hypotheses were examined in an attempt to account for these observations in connection with the species biology. This deficit probably is due to the breeding structure of the population (inbreeding and assortative mating) coupled with limited dispersal ability.     

Diffusion of pulmonary Nematoda in the steinbock (Capra ibex) and the chamois (Rupicapra rupicapra) at the Gran Paradiso National Park and the di Valdieri Reservation]

The results obtained in a survey on the diffusion of pulmonary nematodes in 13 steinbocks (Capra ibex) and 71 chamois (Rupicapra rupicapra) of the Parco Nazionale del Gran Paradiso, and 49 chamois of the Riserva di Valdieri, Italian Western Alps, are reported. The following species of nematodes were identified: Spiculocaulus austriacus, Protostrongylus rufescens, Protostrongylus hobmaieri in steinbocks; Protostrongylus rupicaprae, Protostrongylus hobmaieri, Protostrongylus refescens,Protostrongylus sp., Spiculocaulus austriacus and Dictyocaulus filaria in chamois. The study of two species, viz. Spiculocaulus austriacus and Protostrongylus sp., is still in progress because these nematodes show morphological peculiarities never previously described.     

Isolation of microsatellite loci from the endemic and endangered Adriatic sturgeon (<Emphasis Type="Italic">Acipenser naccarii</Emphasis>)

With the aim of elaborating a breeding plan on a captive stock of the highly endangered Adriatic sturgeon (Acipenser naccarii), a total of 10 polymorphic microsatellite loci were isolated from an enriched library. The results of cross amplification of additional 8 loci previously isolated from A. oxyrinchus, A. fulvescens and Scaphyrinchus platorhynchus are also reported. Given the tetraploid condition of the species the genetic variability was estimated basing on the number of alleles per individuals and the average band sharing.     

Immune responses to Mycoplasma conjunctivae in alpine ibex, alpine chamois, and domestic sheep in Switzerland

The humoral immune response of three alpine chamois (Rupicapra rupicapra rupicapra), two alpine ibex (Capra ibex ibex) and three domestic sheep naturally affected with infectious keratoconjunctivitis (IKC), and four ibex and two sheep experimentally infected with Mycoplasma conjunctivae was analysed. In addition, the local immune response to M. conjunctivae was analysed using conjunctival washes from chamois and sheep. Immunoblot analysis of sera using whole cell antigens of M. conjunctivae revealed the major immunogenic proteins which had molecular masses of 175, 83, 68, 60, 50, 42, 36, and 33 kDa. Major antigens were found at 83, 68, 60, and 42 kDa in both sera and conjunctival washes from naturally infected animals of all three Caprinae species. In experimentally infected animals, antibodies to the 68 and 60 kDa antigens were dominant. Naturally infected animals showed much stronger immune reactions than those experimentally infected, and specific antibodies appeared 2 to 4 wk after experimental infection. To evaluate possible cross-reactions, whole cell antigen of M. conjunctivae was analysed by immunoblot against hyperimmune sera of closely related Mycoplasma spp. Antibodies to the 175, 73, 68, 60, and 33 kDa antigens appeared to be specific to M. conjunctivae. Cross-reactions mainly with 83, 50, and 42 kDa antigens were detected, in particular with M. ovipneumoniae and M. bovoculi hyperimmune sera, but also with antisera against M. capricolum capricolum and M. putrefaciens.