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PubMed2005

In vitro culture and mtDNA fate of ibex-rabbit nuclear transfer embryos.

Rabbit oocyte can be used as the recipient in interspecies somatic cell nuclear transfer (iSCNT). This work was undertaken in order to study the developmental competence of Capra ibex somatic cells reprogrammed by rabbit oocytes and the fate of mitochondria in iSCNT embryos. Metaphase II (MII) oocytes from superovulated rabbit were used as nuclear recipients. The nuclear donors were Capra ibex somatic cells with different proliferative status: population doubling time (PDL) = 15 +/- 2 (group 1), 35 +/- 2 (group 2), 55 +/- 2 (group 3) and 70 +/- 2 (group 4). Oocytes reconstructed with electrical pulses (2.1kV/cm, 10 micros, 2 times) were activated (1.4kV, 20 micros, 2 times) and then cultured in Medium199 containing 10% fetal bovine serum at 38.5 degrees C, 5% CO2 in air. In groups 1, 2, 3 and 4, the fusion rates were 35.83%, 66.03%, 65.40% and 35.35%, respectively. Similar cleavage rates were observed among the four groups. However, the developmental potential to morula/blastocyst from early nuclear donor embryos (16.42%/10.45%) was significantly higher (p < 0.05) than in terminal donor embryos (9.52%/3.81%). Polymerase chain reaction analysis of the mitochondrial (mt) DNA cytb gene demonstrated that mtDNAs from ibex and rabbit could be detected at various developmental stages before implantation. In conclusion, our results provide some original information about rescuing Capra ibex using the iSCNT technique. These results indicate that: (1) enucleated rabbit oocytes make Capra ibex fibroblast nuclei reprogramme; (2) the proliferative status of donor cells affects the efficiency of iSCNT; and (3) rabbit ooplasm rescues the donor-derived mtDNAs, resulting in mtDNA heteroplasmy before implantation.

Animals↗

PubMed2004

Low diversity in the major histocompatibility complex class II DRB1 gene of the Spanish ibex, Capra pyrenaica.

During the last two centuries, the Spanish ibex (Capra pyrenaica) has shown a significant demographic decline as a result of the progressive destruction of its natural habitat, disease epidemics, and uncontrolled hunting. Partial sequencing of the class II MHC DRB1 gene revealed that the Spanish ibex has remarkably low levels of genetic variation at this locus, with only six different DRB1 alleles and an observed heterozygosity of 0.429-0.579. The rates of nonsynonymous vs synonymous substitutions were significantly different in the peptide-binding region (dN/dS=5.347, P=0.002), a feature that indicates that the DRB1 gene is under positive selection. A phylogenetic analysis of the Spanish ibex and a set of domestic goat DRB1 alleles revealed that the reported sequences represent four major allelic lineages. The limited allelic repertoire of the DRB1 gene in the Spanish ibex is likely the direct result of the recent history of population bottlenecks and marked demographic decline of this species. A genetic survey of 13 microsatellite loci was consistent with this idea. The Spanish ibex subspecies C. p. hispanica and C. p. victoriae consistently showed considerably lower levels of microsatellite heterozygosity (Ho=0.184-0.231) and allelic diversity (mean number of alleles per locus=2-2.4) than those reported in other wild ruminants. This study demonstrates the significance of both natural selection and the demographic history of populations in determining patterns of genetic variation at MHC loci. In addition, our results emphasize the importance of locally adapted populations for the preservation of genetic diversity.

Amino Acid Sequence↗

PubMed2005

Relationship between bronchopulmonary nematode larvae and relative abundances of Spanish ibex (Capra pyrenaica hispanica) from Castilla-La Mancha, Spain.

The excretion of bronchopulmonary nematode infective larvae was evaluated in 160 faecal samples of Spanish ibex (Capra pyrenaica hispanica) collected from 13 populations in Castilla-La Mancha, south-central Spain in September 2003. Intensities and prevalences were compared with pasture availability, abundances of wild and domestic ungulates at both levels, i.e. for populations and for faeces in a two-step procedure. Protostrongylid larvae showed similar infection rates (mean intensity: 1.56+/-0.12, n=94; mean prevalence: 25.62+/-6.86%, n=160) to Dictyocaulus spp. (mean intensity: 1.03+/-0.11, n=48; mean prevalence: 30.00+/-7.11%, n=160). At the population level, positive correlations were found between the prevalences of both bronchopulmonary taxa. The prevalence in both groups, but not intensity, also correlated positively with Spanish ibex abundance indexes both for the populations and individual faeces. These findings suggest that: (i) parasite spreading across Spanish ibex populations in Castilla-La Mancha could respond to host density-dependent processes; and (ii) these populations may have similar exposition and/or susceptibility to both bronchopulmonary taxa resulting in similar host-parasite patterns, despite their different life cycles. Bronchopulmonary outputs in the Spanish ibex from Castilla-La Mancha seems not to represent a health risk for this endemic wild ungulate but may be useful in any health surveillance scheme for the increasing populations of Spanish ibex.

Animals↗

PubMed2005

Birth of live Spanish ibex (Capra pyrenaica hispanica) derived from artificial insemination with epididymal spermatozoa retrieved after death.

As a consequence of increasing limitations to maintaining genetic variability in endangered wildlife species, methods of assisted reproduction widely used in domestic animals are being applied to nondomestic species. However, practical efforts have met limited success to date. The Spanish ibex (Capra pyrenaica hispanica) is a wild caprine originating exclusively in the mountains of Spain. This study was designed to evaluate the fertilizing capability of cryopreserved Spanish ibex epididymal spermatozoa recovered postmortem. For this purpose, we have previously evaluated the effect of time elapsed between death and sperm recovery on spermatic parameters, and the fertilization ability of frozen-thawed spermatozoa using heterologous in vivo fertilization by intrauterine insemination in domestic goat (Capra hircus). The time of death significantly affected most sperm quality parameters (motility, viability and intact acrosomes). The fertility obtained by heterologous artificial insemination was 18.7%, and only goats inseminated with spermatozoa recovered within 8h after death became pregnant. Our findings showed that heterologous in vivo fertilization is a useful method to evaluate the fertilizing capacity of sperm samples in rare or wild species. Sperm samples, with verified fertilization ability in the previous trial, were used to inseminate a total of six ibex females. Inseminations resulted in one pregnancy. The study demonstrated for the first time the feasibility of applying artificial insemination in Spanish ibex.

Animals↗

PubMedDate not supplied

Seasonal ovulatory activity and plasma prolactin concentrations in the Spanish ibex (Capra pyrenaica hispanica) maintained in captivity.

Seasonal changes in ovulatory activity, assessed by the measurement of plasma progesterone and plasma prolactin concentrations were monitored in 10 Spanish ibex females captured in the National Wildlife Reserve of Sierra Nevada (South Spain, 37 degrees N). Five of the 10 female ibex showed ovulatory activity with a mean (+/- s.e.m.) duration of the oestrous cycle of 19.4 +/- 1 days (range: 17-23 days). On average, the five cyclic females weighed 28 +/- 0.9 kg. Progesterone cycles occurred only in animals older than 4 years of age. Ovulatory activity extended from December to January. The duration of the breeding season was 43.2 +/- 7.7 days. Ibex females younger than 4 years of age had a body weight lower (P < 0.01 ) than that of adults and none of them displayed ovulations. Plasma concentrations of prolactin levels were significantly affected by season (P < 0.05), following a trend that was roughly parallel to daylength. The highest values occurred in the spring (119.7 +/- 21.4 ng x mL(-1)) and the summer (139.3 +/- 19.8 ng x mL(-1)), the lowest values in the autumn (26 +/- 7.4 ng x mL(-1)) and in the winter (19.7 +/- 3.2 ng x mL(-1)). Our results showed a very restricted breeding season, despite the fact that the Spanish ibex originates from and lives in a temperate latitude, revealing a remarkably good adaptation to the harsh climatic and nutritional conditions of their mountainous habitat.

Age Factors↗

PubMed2005

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.

Animals↗

PubMed1997

Lesions associated with infectious keratoconjunctivitis in alpine ibex.

Following a severe infectious keratoconjunctivitis (IKC) epizootic in free-ranging alpine ibex (Capra ibex ibex) in Switzerland in 1993, 19 animals were examined from six different populations. Mucopurulent exudates, reddened conjunctiva and mononuclear inflammatory cell infiltration in the conjunctiva and the limbic area were observed in mild cases. In more severe cases, lesions ranged from perilimbic neovascularization to corneal edema, erosion, ulceration and neovascularization accompanied by infiltration of neutrophils. Sometimes an iridocyclitis was observed. In the most advanced stages, the cornea was perforated and an anterior synechia was present. Lesions found in ibex affected with IKC indicated a non-generalized, specific ocular disease. The other organs investigated did not show alterations suggestive of changes induced by agents which might cause IKC, including Chlamydia spp. and Mycoplasma. spp. The microbiological findings indicate that Mycoplasma conjunctivae is the primary pathogenic agent causing IKC in this species in Switzerland.

Animals↗

PubMed1998

Isolation of Brucella melitensis from alpine ibex.

Eleven alpine ibex (Capra ibex) and 27 chamois (Rupicapra rupicapra) from Gran Paradiso National Park (Italy) were examined in March 1996. A 7-yr-old ibex buck had thick-walled carpal joints and enlargement of the right testicle characterized by necrosis and fibrosis. Microscopically, testicular lesions were characterized by large areas of necrosis, fibrosis with irregular aggregates of macrophages and lymphocytes, and scattered foci of suppuration. Specimens of the carpal bursae and testicle were cultured in serum dextrose agar and serum dextrose antibiotic plates. A Gram-negative coccobacillus was isolated from the testicle and subsequently identified as Brucella melitensis biotype 2. This is the first recognized case of brucellosis in alpine ibex.

Animals↗

PubMed1999

Sarcoptic mange in Spanish ibex from Spain.

The Spanish ibex (Capra pyrenaica hispanica) population of the "Sierras de Cazorla, Segura y Las Villas" Nature Park (Spain) was isolated as the result of a severe epidemic of sarcoptic mange. In this context, the dynamic characteristics of the disease were analyzed in a wild group consisting of 35 animals from the beginning of the epizootic (when the mating period started) to the extinction of the population due to mange. Monthly tracking permitted the sequential characterization of the pathology in each animal. The duration of the disease was 2 to 3 mo, evolving to severe disease and terminating in death. Incidence and prevalence rates in terms of morbidity and severity, and mortality and lethality were calculated. At the end of the mating season, 81% of the population were affected. There were no statistically significant differences in severity of the disease across sex or age categories of the animals. Most of the carcasses were found in caves used as refuge and/or near rivers or streams. Additionally, 46 of the 63 (73%) ibex captured in different areas of the nature park were naturally infected with the Sarcoptes scabiei. Infected ibex were examined for number of mites during the initial stage of the disease (n = 3), in the development stage (n = 12), in the consolidation stage (n = 17), and in the chronic stage (n = 14). The prevalence of mites in different anatomical regions was determined in each of these phases of the infection. A histological study of the skin lesions was conducted in 22 animals. Both the clinical and the pathological (macroscopic and microscopic) aspects of the sarcoptic mange in Spanish ibex corresponded to the classic description of sarcoptic mange in other wild and domestic small ruminants.

Age Factors↗

PubMed2000

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.

Animals↗

PubMed2006

Horn growth related to testosterone secretion in two wild Mediterranean ruminant species: the Spanish ibex (Capra pyrenaica hispanica) and European mouflon (Ovis orientalis musimon).

Seasonal variations in the horn development and testicular activity of the Spanish ibex (Capra pyrenaica hispanica) (n=6) and European mouflon (Ovis orientalis musimon) (n=5) were monitored to determine the role of increasing testosterone concentration on the arrest of horn growth during the rutting season. Marked seasonal variations in the rate of horn growth (P<0.01) and testicular activity (P<0.001) were seen in both species, although the magnitude and timing of these changes were different (P<0.01). Horn growth rate was inversely correlated to seasonal levels in testosterone plasma concentration in both species (ibex: R=-0.45, P<0.01; mouflon R=-0.51, P<0.01). In the mouflon, the increase in plasma testosterone concentration recorded in September (P<0.05 compared with the lowest concentration) coincided with a significant reduction in horn growth (P<0.05). In the ibex, the increase in plasma testosterone concentration in October (P<0.05 compared with the lowest concentration) was associated with a significant arrest of horn growth in November (P<0.05). These results appear to support the hypothesis that high peripheral plasma levels of testosterone are linked with the seasonal arrest of horn growth during the rutting period.

Animals↗

PubMed2006

Sarcoptic mange and metapodial development in growing male Iberian ibex (Capra pyrenaica).

The effect of mange (Sarcoptes scabiei) on metapodial growth was studied in 24 uninfested and 16 infested male Iberian ibex from Sierra Nevada National Park (southern Spain). Our results show that infested ibex had smaller metatarsal bones, lower body weight, and a delayed rhythm of ossification, by about 1 year. We conclude that skeletal development and body weight in the Iberian ibex are sensitive to sarcoptic mange.

Animals↗

PubMed2002

An evaluation of the application of infrared thermal imaging to the tele-diagnosis of sarcoptic mange in the Spanish ibex (Capra pyrenaica).

The application of infrared thermal imaging to the diagnosis of sarcoptic mange in the wild Spanish ibex (Capra pyrenaica) was evaluated. Seventy-three ibexes (35 males, 38 females) of varying ages were studied. Each animal was observed using conventional binoculars (OT) to detect lesions characteristic of mange. Infrared thermography (IR) was then performed and the resultant image judged negative or positive. The distance from the thermograph to the animal was measured, and the animal killed. Skin samples were taken for mite detection by routine laboratory diagnosis (LAB). The most sensitive and specific technique for the tele-diagnosis of sarcoptic mange in the Spanish ibex is OT, as it permits diagnosis over greater distances than IR, which sensitivity is impaired at distances >100m. When disease prevalence is low, such as in initial and final phases of an epidemic, a more sensitive technique would be valuable in detecting all affected animals.

Aging↗

PubMed1998

Seroprevalence of Babesia ovis in Spanish ibex (Capra pyrenaica) in Catalonia, northeastern Spain.

A serologic survey was carried out in order to detect antibodies against Babesia ovis in a large population of Spanish ibex (Capra pyrenaica) from a hunting reserve in Catalonia, northeastern Spain. For this purpose, an indirect fluorescent antibody test (IFAT) was developed using a B. ovis isolate of ovine origin as antigen. Of the total 475 sera tested, 155 (32.6%) showed titres between 1:160 and 1:1280 and were considered positive. These results reveal that exposure of Spanish ibex to B. ovis is common in the studied area. No significant differences could be detected when comparing season or year of capture and age or sex of the animals in positive and negative samples. A high proportion of low titres was found in comparison to those reported by other researchers in sheep in Spain; this could be a consequence of the existence of some minor antigenic differences between B. ovis of domestic sheep and that found in Spanish ibex.

Age Distribution↗

PubMed2004

The fate of mitochondria in Ibex-hirus reconstructed early embryos.

Inter-species nuclear transfer could be used to preserve North Goat (Capra ibex), an endangered species. We established the culture conditions for ibex-hirus reconstructed embryos and optimized the method for DNA extractions of a single cell and early cloned embryo. By using mitochondria-specific probes of ibex and hirus respectively we found that mitochondria of donor cells can co-exist with recipients in 1-cell and 2-cell stages of the reconstructed embryos but not in the following developmental stages.

Animals↗

PubMed2026

Spatial proteomic mapping of the human and mouse retina using IBEX.

We generated a comparative spatial proteomic atlas of the human and mouse retina using a highly multiplexed immunohistochemistry technique called iterative bleaching extends multiplexity (IBEX). We refined the IBEX workflow by integrating an antibody dissociation option alongside chemical bleaching. This dual strategy enabled removal of the entire antibody complex, permitting the flexible use of antibodies from the same host species across iterative cycles. We coupled this workflow with super-resolution imaging via deconvolution and applied it to the retina of healthy humans and WT mice and the Crb1rd8 mouse model. We successfully imaged over 25 protein markers on human and mouse tissue sections, generating spatial atlases of the major retinal cell populations. Cross-species protein expression was compared to scRNA-seq datasets to identify protein and transcript disparities. Super-resolution IBEX delineated the ultrastructural features of the outer limiting membrane (OLM), identifying CD44 as a core structural component tightly colocalized with a highly organized F-actin belt within Müller glial endfeet. Using the Crb1rd8 mouse model, disruption of this complex was spatially associated with rosette formation and OLM structural failure. In summary, spatial proteomic atlases of the human and mouse retina were used to reveal insights into the arrangement of major retinal cell populations and OLM structure.

Animals↗

PubMed2002

Infectious keratoconjunctivitis of ibex, chamois and other Caprinae.

Infectious keratoconjunctivitis (IKC) caused by Mycoplasma conjunctivae is a highly contagious ocular infection which is common in domestic sheep and goats. In the European Alps, IKC is often observed in Alpine chamois (Rupicapra r. rupicapra) and in Alpine ibex (Capra i. ibex), but the disease has also been described in other wild Caprinae in the Pyrenees and in New Zealand. The infection is characterised by inflammation of the conjunctiva and cornea, and in the most advanced stages, the cornea is opaque or even perforated. In IKC outbreaks in chamois and ibex, spontaneous recovery is the most prevalent outcome of the disease. However, mortality can occasionally reach 30%. Recent studies in eastern Switzerland indicated that M. conjunctivae infection is not self-maintained in chamois. In contrast, the disease is endemic and self-maintained in the domestic sheep population. Spillover of the agent from sheep living in proximity during summer may be the origin of point-source epidemics in wild Caprinae. Flies are likely to play a central role in interspecific transmission of M. conjunctivae on alpine meadows. When outbreaks of IKC occur in wildlife, a special effort should be made to avoid unnecessary human disturbance in affected areas. However, animals presenting irreversible ocular lesions should be shot by professional gamekeepers to prevent suffering. Prevention of IKC in wild Caprinae should focus on preventing the spillover of M. conjunctivae from livestock. However, studies are required to evaluate the distribution of M. conjunctivae infection in domestic sheep in several countries and to assess the risk of IKC spillover from domestic animals to wildlife. In addition, immunological studies should be performed to develop tools which could lead to the control of M. conjunctivae infection in domestic sheep.

Animals↗

PubMed2001

Experimental ivermectin treatment of sarcoptic mange and establishment of a mange-free population of Spanish ibex.

Ivermectin was used to treat sarcoptic mange in Spanish ibex (Capra pyrenaica hispanica). Its therapeutic effectiveness was analyzed when it was administered through subcutaneous injection, to sick animals in the consolidation stage of mange (third phase) and, with double injections to chronically affected animals (fourth phase) at a dosage of 0.2 or 0.4 mg/kg body weight (bw). Three wk after treatment, the animals in the third phase of mange treated with a high dose (0.4 mg/kg bw) of ivermectin were completely cured. The same result was achieved after 4 wk of treatment in those animals in phase 3 of mange when 0.2 mg/kg body weight was used. Double injection with ivermectin, even at high doses, did not guarantee the complete cure of all cases of sarcoptic mange in the chronic stage (phase 4); only three of six animals were free of Sarcoptes scabiei. The second experiment consisted on the application of a sanitation program in order to obtain a population of Spanish ibex free from S. scabiei, starting with free-ranging animals, some of them healthy and others sick. After capture the animals were classified as chronically ill, in which case they were excluded from the program, mite carriers and healthy specimens. All the animals were treated first topically with foxim (500 mg/l) and subcutaneously with ivermectin (0.4 mg/kg bw). The infected animals were housed in the treatment pen, and received two doses of ivermectin (0.2 mg/kg bw) at an interval of 15 days, then spent 15 days in the quarantine pen, where they received a further dose before they were included in the pool of healthy animals, and immediately were placed in the quarantine phase. The sanitation we implemented was fully effective in curing the affliction of Spanish ibex affected by S. scabiei.

Animals↗