Anatomy and computed tomography of the nasal cavity, nasal conchae, and paranasal sinuses of the endangered Patagonian huemul deer (Hippocamelus bisulcus).

This study explores for the first time the shape, volume, and configuration of nasal cavity structures of the endangered Patagonian huemul deer via computed tomography (CT). Three-dimensional (3D) reconstructions derived from data sets obtained from five Patagonian huemul deer skulls were analyzed. Using semiautomatic segmentation, 3D models were created of all the sinus compartments and nasal conchae. Volumetric measurements were taken of seven sinus compartments. The Patagonian huemul deer has a wide, large nasal cavity, with an osseous nasal aperture typical of cervids and a choana with characteristics that differentiate it from the pudu and roe deer. It also has six nasal meatuses and three nasal conchae, with the ventral nasal concha having the greatest volume and surface, which given its extension ensures a greater ability to humidify and heat the air. Further analysis showed the complex system of paranasal sinuses to be characterized by a rostroventral and interconnected group, where communication with the nasal cavity is common through the nasomaxillary opening, and a caudodorsal group that communicates with the nasal cavity through openings in the nasal meatuses. Our study of the endangered Patagonian huemul deer documents an intricate, and in some nasal cavity structures, unique morphological construction which may predispose it to higher rates of sinonasal afflictions due largely to its nasal complex anatomy, thus affecting its high cultural value.

A Protocol to Assess the Welfare of Patagonian Huemul (Hippocamelus bisulcus) in Conservation Centers.

Animal-welfare-assessment protocols are important for identifying welfare problems in conservation programs. This study aimed to develop a baseline welfare protocol for the assessment of the welfare of Patagonian huemuls (Hippocamelus bisulcus) in conservation centers. This protocol is based on the Welfare Quality® (WQ) framework for cattle and was developed with the consideration of the available research on the behaviors, ecology, conservation, and captive management of this species, as well as welfare-assessment protocols for other ungulate species. As a result, the protocol was specifically developed for Patagonian huemuls and included four principles, 12 criteria, and 23 animal- and resource-based indicators. The twelve criteria of the WQ protocol were reduced to nine, and three new criteria were added because they were both feasible and essential for welfare assessment in captive Patagonian huemuls. This protocol is mainly intended to identify welfare problems in endangered species in the context of conservation centers (reproduction, rescue, rehabilitation, or treatment centers). Thus, the aggregation of different measures to obtain a global score was not proposed. However, a scoring system that assigns a value on a 0-2 scale (0 = no welfare concern; 1 = welfare concern; 2 = urgent welfare concern) was proposed for each category. Although further research is still needed to fully validate the protocol, this is the first development of a protocol to assess Patagonian huemul welfare, and it can be used as a basis for the development of welfare-assessment protocols for other deer species in captivity.

Molecular and phylogenetic analysis of herpesviruses in endangered free-ranging cervids of Chile: ovine gammaherpesvirus-2-A novel threat to wild and domestic animal health in Chilean Patagonia.

Introduction: Herpesvirus infections have been highlighted as emerging diseases affecting wildlife health and the conservation of several taxa. Malignant catarrhal fever (MCF) and infectious keratoconjunctivitis (IKC) are two viruses that infect wild ruminants. Nevertheless, epidemiological data on herpesviruses in South American wild ruminants are limited. An outbreak of caprine gammaherpesvirus-2 (CpHV-2) that recently was suspected as the cause of MCF in southern pudus (Pudu puda) prompted the need to conduct molecular screenings in Chilean cervids to understand the epidemiology of herpesviruses. The aim of this study was to determine the occurrence and genetic diversity of herpesviruses in free-ranging cervids from Chile. Methods: Herpesvirus infection was assessed in antemortem blood samples (n = 86) from pudus (n = 81) and huemuls (Hippocamelus bisulcus) (n = 5), as well as postmortem samples of spleen (n = 24) and lung (n = 3) from pudus, using a nested pan-herpesvirus PCR assay. Results: Combining all suitable sample types, DNA of pudu gammaherpesvirus-1 was detected in five pudues and five huemuls, with an overall prevalence of 9.90% (n = 10/101; 95% CI = 5.11-17.87%). One pudu tested positive for ovine gammaherpesvirus-2 (n = 1/96; 1.04%; 95% CI = 0.05-6.49%), and one pudu tested positive for a Macavirus sequence with 98.63 similarity to ovine gammaherpesvirus-2 (n = 96; 1.04%; 95% CI = 0.05-6.49%). Discussion: To the best of our knowledge, this is the first report of a herpesvirus in huemul and of ovine gammaherpesvirus-2 in Chile. Our results also confirm the active circulation of herpesvirus in free-ranging cervids in Chilean Patagonia, and as such, MCF should be considered as a possible cause of disease in free-ranging Chilean pudus and livestock species. Further research is necessary to develop a plan of systematic monitoring (serological and pathological screening) of herpesviruses in Chilean wild and domestic ruminants to understand their diversity and impact on animal health and conservation.

Skull anatomy of the endangered Patagonian huemul deer(Hippocamelus bisulcus).

Studies of the normal skull anatomy of the Patagonian Huemul deer are scarce. Currently, the findings of bone lesions in the skull associated with metabolic imbalances are frequent in the literature. The objective of this study was to provide anatomical and morphometric data of the cranium and facies including a morphofunctional interpretation as a reference for clinical, ecomorphological and educational purposes. Five skulls were described, measured, scanned and digitally reconstructed. The presence of a caudal projection of the vomer bone, the absence of the facial tubercle and thin bones forming the cranial cavity were observed. Linear measurements allowed the skull to be classified as dolichocephalic, hyperlepten and ultra-dolichocranial. In conclusion, the Patagonian Huemul has a long head, an extremely long skull and a very narrow face. The thickness of the bones that made up the walls of the cranial cavity suggests chronic metabolic imbalances in response to mineral deficiency. The anatomical and morphometric data obtained in this study strongly emphasizes the relevance of considering the implementation of such management policies that aim at promoting an optimal nutritional context.

Partially Reversible Immobilization of Free-Ranging Huemul Deer (Hippocamelus bisulcus) with Medetomidine-Ketamine and Atipamezole.

A combination of intramuscular medetomidine and ketamine was used to immobilize 46 free-ranging huemul deer (Hippocamelus bisulcus) with a remote drug delivery system in Chilean Patagonia for tagging and biological sampling. Captures occurred in May-October of 2005-09 between fall and early spring in the southern hemisphere. An initial dose of 6.6 mg medetomidine and 185 mg ketamine was adjusted after 17 captures to 3 mg and 200 mg, respectively, in the 29 remaining deer. Mean±SD adjusted dose was 0.042±0.012 mg/kg of medetomidine and 2.929±0.427 mg/kg of ketamine. Inductions were calm and the mean±SD time to sternal recumbency was 10.3±10.1 min. Palpebral reflex and jaw tone were present during immobilization. Atipamezole at 5 mg/mg of medetomidine was administered intramuscularly for reversal after 55.3±18.8 min procedure time. Recoveries were smooth and mean±SD time to standing was 10.2±3.3. All immobilized animals were hypoxemic by pulse oximetry (blood oxygen saturation approximately 81%). Three animals that developed apnea were resuscitated through chest compression and atipamezole administration, another regurgitated during capture, and all developed tachypnea. The combination of medetomidine-ketamine and atipamezole can be used for partially reversible immobilization of huemul, but supplemental oxygen should be administered, blood oxygenation should be monitored, and equipment for intubation and manual ventilation should be available.

Functional limb anatomy in a refugee species: The endangered Patagonian huemul deer (Hippocamelus bisulcus).

Early naturalists already considered huemul rare, refuged and a stocky, short-legged mountain deer, 163 years before declared endangered (1972). Anatomically, huemul do not overlap with rock-climbers previously considered analogous, as corroborated in this paper by including additional huemul cases. Assertions that population declines are caused principally via livestock infections remain unfounded. Instead, osteopathology in multiple populations across 1,000 km, affecting 57% among dead and 86% among live specimens, may relate to micronutrient deficiencies. Historically classified a mountain deer, widespread osteopathology, micronutrient deficiencies and lack of recovery qualify huemul as a refugee species. Recovery strategies thus must include repopulating historical distribution sites.

Nutrition as an etiological factor causing diseases in endangered huemul deer.

OBJECTIVES: Distinct diseases prevent endangered huemul deer (Hippocamelus bisulcus) recovery. Fundamental etiological factors include nutriments, a mayor component of habitat quality. Undernutrition affects growth, skeletal development, osteopathology, reproduction and immunocompetence: this paper amplifies data corroborating micro-nutrient deficiencies among huemul. RESULTS: In Argentina, 57% huemul cadavers exhibited osteopathology, with new cases reported here. Recently, 86% live huemul had osteopathology: cranial lesions involved antemortem tooth loss, reducing feeding efficiency and body condition, with starvation deaths. This population had tissues well deficient compared to other cervids, averaging 0.28 ppm selenium, 4.98 ppm copper, whereas for manganese 55% were deficient (2.52 ppm) and 45% adequate (42.79 ppm). Recently, lesions in one Chilean huemul were interpreted to stem from parapoxvirus. That population also has cases with cranial osteopathologies, high disease susceptibility (parapoxvirus, parasitism, foot lesions), crippled antlers, and low density, indicative of marginal habitat and primary etiological factors like undernutrition and immunosuppression. The reported atypical symptoms attributed to parapoxvirus may relate to probable diagnostic limitations, but does support presence of nutritional deficiencies. Patagonia has selenium deficient plants and livestock, including severe muscular dystrophy, and soil levels in extant huemul areas considered very deficient. Moreover, 73% of Chilean huemul were selenium deficient and 64% severely deficient with concomitant cranial osteopathology.

Morphometric, anatomic and radiographic study of bone of the pelvic limb of endangered Patagonian huemul deer (Hippocamelus bisulcus) / Estudio morfométrico, anatómico y radiográfico de los huesos del miembro pélvico del huemul patagónico en peligro de extinción (Hippocamelus bisulcus)

Currently, the Patagonian huemul (Hippocamelus bisulcus) is endangered. Anatomical studies focused on understanding the anatomy of the deer most threatened by extinction in South America is a challenge for anatomists, veterinarians and biologists. Research carried out for its conservation has focused mainly on its ecology and pathology, leaving gaps in anatomical knowledge, which is basic and important for a comprehensive understanding of this species. Gross anatomy and radiography of the skeleton of the pelvic limb was performed in three adult Patagonian huemul. Bone specimens of three skeletally mature Patagonian huemul deer were used for gross osteological and radiographic studies. This study was conducted to reveal the morphometric and morphological features of the ossa membri pelvini of the Patagonian huemul. The main findings suggest the presence of powerful extensor muscles in the coxofemoral, femorotibial and tarsal joints, useful during walking, jogging and propulsion in deer. In general, the MNF of Patagonian huemul differ in position with respect to domestic ungulates. In addition, the presence of a notch or obturator canal was observed, with variability in morphology and development among the specimens.

Actualmente, el Huemul Patagónico (Hippocamelus bisulcus) está en peligro de extinción. Los estudios morfológicos centrados en comprender su anatomía han sido un desafío para morfólogos, veterinarios y biólogos. Investigaciones enfocadas en su conservación, se han centrado principalmente en su ecología, patología y en la descripción de sus principales agentes infecciosos, dejando vacíos en el conocimiento anatómico básico, y por ende, su comprensión morfofisiológica. El objetivo de este estudio fue describir la anatomía, morfometría y radiología macroscópica normal y detallada del ossa membri pelvini en tres especímenes de Huemul Patagónico, incluida una interpretación funcional como referencia para uso clínico y conservacionista, investigación biomédica y fines de enseñanza. Los principales hallazgos sugieren la presencia de potentes músculos extensores en las articulaciones coxofemoral, femorotibial y tarsal, útiles durante la marcha, el trote y la propulsión en los ciervos. En general, los forámenes nutricios principales del Huemul patagónico difieren en posición con respecto a los ungulados domésticos. Además, se observó la presencia de una muesca o canal obturador, con variabilidad morfológica y de desarrollo entre los especímenes.

Anatomy, Morphometry and Radiography in the thoracic limb bones of the Patagonian Huemul Deer (Hippocamelus bisulcus).

The aim of this study was to provide morphometric, anatomic and radiographic data of the thoracic limb bones of the Patagonian Huemul (Hippocamelus bisulcus) including a functional interpretation of this, as a reference for clinical use, biomedical research and teaching purposes. Currently, the Patagonian huemul deer is in danger of becoming extinct due to multiple causes. Research carried out for its conservation has focused mainly on its ecology and pathology, leaving gaps in biological knowledge, which is basic and important for its comprehension. This study was conducted to reveal the gross osteology and radiology features of the thoracic limb bones of the Patagonian huemul deer. The osteological findings suggest the presence of powerful flexor muscles in the scapulohumeral and elbow joints, useful to cushion the jumps. Also, the principal nutrient foramen of Patagonian huemul differs in position with respect to domestic ungulates, which may be important to consider during surgical procedures. Finally, the radiographic data can provide new information about the tissue loading conditions in Patagonian huemul, so that this new knowledge can be of great importance for a better understanding of mechanically induced or adaptive changes in bone produced by habitat or other ecological phenomena.

Elusive cranial lesions severely afflicting young endangered Patagonian huemul deer.

OBJECTIVES: Most subpopulations of endangered huemul deer (Hippocamelus bisulcus) fail to recover, frequently due to osteopathology. Equivalent pathology was detected only postmortem in an additional deer 365 km further north, stressing the need to improve clinical evaluations of live huemul. RESULTS: Captured on a farm and attended by authorities in charge of huemul, the deer was considered apt for relocation and release. Delays with attendance and lack of reversal drugs resulted in his death. The subsequent necropsy revealed severe osteopathology particularly in mandibles and maxillae. Such disease in another southern population affected 57+ % among dead adults, and 86% among live adults. The present case stems from a new subpopulation, isolated 365 km further north. Such severe pathology demands that individuals be rehabilitated, especially relevant with severely endangered species, because liberations will cause premature death and loss of reproductive lifetime. Live huemul must be examined utmost professionally especially regarding this pathophysiognomy. This incidence represents the typical situation of extant huemul, being displaced from their traditional migratory behavior to utilize fertile low-elevation habitat. This young male may have been dispersing, but reaching valleys usually leads to death due to locally intense anthropogenic activities.

Troubling disease syndrome in endangered live Patagonian huemul deer (Hippocamelus bisulcus) from the Protected Park Shoonem: unusually high prevalence of osteopathology.

OBJECTIVE: The last 1500 endangered Patagonian huemul deer (Hippocamelus bisulcus) exist in > 100 groups which are not recovering. Prevalence of osteopathology in dead huemul was 57+% (Argentina), whereas similar cases in Chile were accompanied by selenium deficiency. The first clinical cases from live wild huemul confirm widespread osteopathology which explains short life spans, low recruitment, and thus absence of population recovery. RESULTS: The first-ever radio-collaring of 3 male huemul in Argentina and 3 females, plus a fresh female carcass allowed examination of 7 huemul. Of these, 86% were diseased and clinical pathophysiognomy included lameness, affected hoof, exfoliation of 2-7 incisors, other cranial osteopathologies, and muscle atrophy. The parsimonious explanation for absent population recovery is high prevalence of osteopathology as evidenced earlier in carcasses, and now by these clinical cases. Areas currently used by huemul have reduced selenium bioavailability, very deficient soil levels, and overt selenium deficiency in local livestock and plants. These areas are known to result in primary iodine deficiency which is aggravated by selenium deficiency. The nexus to nutritional ecology of huemul likely is inaccessibility to most fertile lowlands and traditional winter ranges, elimination of migratory traditions, and concomitant elimination of source populations.

Corynebacterium pseudotuberculosis Infection in Patagonian Huemul ( Hippocamelus bisulcus ).

Corynebacterium pseudotuberculosis is an intracellular bacteria and the etiologic agent of caseous lymphadenitis in domestic and wildlife species. We report C. pseudotuberculosis infection in Patagonian huemul ( Hippocamelus bisulcus ) from the Cerro Castillo National Reserve, Region of Aysen, Chile. Subcutaneous abscesses in the abdominal and pectoral regions from two animals were sampled and bacteriologic isolation was performed. In both cases, we isolated a C. pseudotuberculosis strain belonging to the ovine genotype. In addition, one isolate was resistant to ciprofloxacin and streptomycin. We report that H. bisulcus is a susceptible species to this bacterium, which is transmitted by direct or indirect contact with domestic sheep ( Ovis aries ) and which represents a potential conservation threat to populations of H. bisulcus . Additional research and prevention efforts should be addressed.

Attempts at Applying Cloning to the Conservation of Species in Danger of Extinction / Intentos de Aplicar la Clonación en la Conservación de Especies en Riesgo de Extinción

The somatic cloning by transfer of the nuclei of differentiated adult cells to previously enucleated oocytes is a promising technique for the production of embryos of high genetic value. The better mastering of somatic cloning gives us the possibility to produce embryos from endangered species. The huemul is an Andean native deer, that has been declared an endangered species, it holds a great patrimonial value and it is a Chilean national emblem. In Chile the huemul has the status of protected animal on thirteen Parks and National Reserves managed by Corporacion Nacional Forestal (CONAF). This protection, however, is considered insufficient due to the few geographical connections between the different protected areas. Furthermore, a great proportion of these areas are not subjected to use or they do not constitute adequate habitats. Many authors have proposed that the use of biotechnological methods in reproduction and assisted procreation may help conservational programs orientated to the protection of deer species threatened by extinction. All the anterior prompted us to initiate this study concerning the production of cloned huemul embryos.

El clonaje somático por transferencia del núcleo de células diferenciadas adultas a un ovocito, al que se le ha extraído el núcleo (enucleado), es una técnica prometedora para la producción de embriones de alto valor genético. El mejor dominio del clonaje somático da la posibilidad de producir embriones de especies amenazadas de extinción. El huemul es un ciervo andino autóctono, declarado como especie en peligro de extinción. tiene un gran valor patrimonial, y es emblema de la nación chilena. En este país, el huemul se encuentra protegido en trece Parques y Reservas Nacionales, manejadas por la Corporación Nacional Forestal (CONAF). Sin embargo, su protección se considera insuficiente debido a la baja conectividad entre las áreas protegidas y además, una gran proporción de estas áreas no son utilizadas o no constituyen un hábitat adecuado. Para las especies de cérvidos en vías de extinción el uso de biotecnología reproductiva y métodos de procreación asistida, según varios autores, pueden ayudar a los programas de conservación. Las técnicas clásicas de producción de embriones basados en superovulación, inseminación artificial y transferencia embrionaria,en los cérvidos, han resultado muy dificultosa. Esto, sumado a las características del huemul, que no permite su estabulación en cautiverio, nos ha movido a iniciar un estudio tendiente a la producción de embriones clonados de esta especie.