Exposure to disease agents in the endangered Iberian lynx (Lynx pardinus)

The Iberian lynx (Lynx pardinus) is the most endangered felid species in the world. Lynx populations have decreased dramatically in size and distribution in the last four decades, thus becoming increasingly vulnerable to catastrophic events such as epizooties. From 1989 to 2000, serum samples were obtained from 48 free-ranging lynx captured in the Doñana National Park (DNP, n?=?31) and mountains of Sierra Morena (SM, n?=?17) in southern Spain. Samples were tested for antibodies against Toxoplasma gondii, feline herpesvirus 1 (FHV-1), feline calicivirus (FCV), feline/canine parvovirus (FPV/CPV), feline coronavirus, feline immunodeficiency virus (FIV), feline leukaemia virus and canine distemper virus (CDV) and for FeLV p27 antigen, to document baseline exposure levels. Antibodies against T. gondii were detected in 44% of lynx, with a significantly greater prevalence in DNP (61%) than in SM (12%). In DNP, prevalence was significantly higher in adult (81%) than in juvenile and sub-adult (41%) lynx, but no such difference was observed in SM. Low prevalences (≤11%) of minimally positive titres were found for FHV-1, FCV and FPV/CPV. This, combined with the lack of evidence for exposure to CDV, FIV and FeLV, suggests that these lynx populations are naïve and might be vulnerable to a disease outbreak in the future. Because of the reduced size of lynx populations, the documented low level of genetic variation (particularly in the DNP population) coupled with the recently documented state of immune depletion in a majority of necropsied lynx, it is important to better understand the threat and potential impact that disease agents might pose for the conservation of this endangered species. Future surveillance programs must include possible disease reservoir hosts such as domestic cats and dogs and other wild carnivores.     

Helminth parasites of the endangered Iberian lynx (Lynx pardinus) and sympatric carnivores

Five critically endangered Iberian lynxes (Lynx pardinus) and 35 other sympatric carnivores (19 feral cats Felis catus, 12 Egyptian mongooses Herpestes ichneumon, and 4 common genets Genetta genetta) were analysed for helminths in Sierra Morena and Do?ana area (southern Spain). Ancylostoma tubaeforme, which was believed to be harmful for lynx cubs according to a previous study, was present in the only lynx and in 53% of cats analysed in Do?ana (80% in adult cats). Other species shared in both areas were Toxocara cati (1 lynx, 31% of cats), Joyeuxiella pasqualei (1 lynx, 21% of cats) and Mesocestoides sp. (2 lynxes, 5% of cats). Only one mongoose was parasitized, harbouring larvae of two acantocephalan species not previously reported in the Iberian peninsula (Centrorhynchus (Sphaerirostris) lancea and Centrorhynchus (Longirostris) undulatus). Feral cats may be a reservoir for hookworms and other helminths affecting the Iberian lynx. In contrast, mongooses and genets may not play a role in the epidemiology of these species.     

Ectoparasites of the Irish stoat

A sample of 196 Irish stoats Mustela ermineae hibernica Thomas & Barrett-Hamilton (Carnivora: Mustelidae) were examined and yielded 2580 arthropod ectoparasites, including 1819 larval mites Neotrombicula autumnalis (Shaw) from a single host. Other ectoparasites recovered included lice (99% Trichodectes ermineae (Hopkins], ticks (mostly Ixodes hexagonus Leach) and four species of fleas. The ticks and fleas were considered to have come mainly from nests of other hosts. The flea species did not reflect the status of their usual hosts in the diet of stoats, but did reflect the stoats' use of their usual hosts' habitat.     

Management measures to control a feline leukemia virus outbreak in the endangered Iberian lynx

The feline leukemia virus (FeLV) is a retrovirus that affects domestic cats all over the world. Its pathogenic effects generally include anemia, immunosuppression or tumors. Dissemination over populations is linked to cat sociality, because the virus is transmitted by direct contact. Although the domestic cat is its common host, FeLV infection has also been described in some wild felids. In the Iberian lynx Lynx pardinus , some sporadic FeLV infection cases have been reported since 1994, but an outbreak with the involvement of several animals has never been described until now. During spring 2007, an FeLV outbreak hit the Doñana (SW Spain) population. The infection rapidly spread throughout the densest subpopulation throughout Doñana. Infected animals showed very acute anemic disease, most of them dying in <6 months. To avoid FeLV dissemination, a control program was carried out that included removal of viremic lynxes, vaccination of negative individuals and reduction of the feral cat population. The program was implemented both in Doñana and in Sierra Morena populations. In Doñana, around 80% of the total lynx population and 90% of the outbreak focus subpopulation were evaluated. Seven out of the 12 infected individuals found died and two reverted to latency; the remaining viremic animals have been kept in captivity. The outbreak appears to have been successfully confined to the subpopulation where the virus appeared and no more cases have been found since August 2007. In the larger Sierra Morena population, 8% of the lynx population was surveyed. Thirty-four uninfected Iberian lynxes were vaccinated at least once. The FeLV prevalence was found to be 27% in the Doñana population and 0% in the Sierra Morena population.     

Detection and identification of Bartonella sp. in fleas from carnivorous mammals in Andalusia,Spain

A total of 559 fleas representing four species (Pulex irritans, Ctenocephalides felis, Ctenocephalides canis and Spilopsyllus cuniculi) collected on carnivores (five Iberian lynx Lynx pardinus, six European wildcat Felis silvestris, 10 common genet Genetta genetta, three Eurasian badger Meles meles, 22 red fox Vulpes vulpes, 87 dogs and 23 cats) in Andalusia, southern Spain, were distributed in 156 pools of monospecific flea from each carnivore, and tested for Bartonella infection in an assay based on polymerase chain reaction (PCR) amplification of the 16 S–23 S rRNA intergenic spacer region. Twenty‐one samples (13.5%) were positive and the sequence data showed the presence of four different Bartonella species. Bartonella henselae was detected in nine pools of Ctenocephalides felis from cats and dogs and in three pools of Ctenocephalides canis from cats; Bartonella clarridgeiae in Ctenocephalides felis from a cat, and Bartonella alsatica in Spilopsyllus cuniculi from a wildcat. DNA of Bartonella sp., closely related to Bartonella rochalimae, was found in seven pools of Pulex irritans from foxes. This is the first detection of B. alsatica and Bartonella sp. in the Iberian Peninsula. All of these Bartonella species have been implicated as agents of human diseases. The present survey confirms that carnivores are major reservoirs for Bartonella spp.     

Proximate and ultimate causes of dispersal in the Iberian lynx Lynx pardinus

Most studies on the causes of animal dispersal focus on speciesof birds or small mammals, but there are few such studies onsolitary carnivores. A complete picture of the causes of animaldispersal is not possible without considering cases on a representativeset of animals. The Iberian lynx is a medium-size, solitarycarnivore that inhabits metapopulations, where dispersal playsa prominent role. Between 1983 and 1998 we studied the proximateand ultimate causes of dispersal in Iberian lynx in the Doñanametapopulation (southwestern Spain), based on radio-trackingof 49 individuals. Saturation of limited breeding areas in thissmall population leads to high dispersal rates in both sexes.Most lynxes dispersed from their natal area between 12 and 24months of age. Males younger than 1 year old tended to dispersein a higher proportion than females of the same age. However,high dispersal rates for both sexes by the age of 2 years andthe lack of differences in the dispersal distances do not supportinbreeding avoidance as an ultimate cause of dispersal. Dispersalmainly started between January and June (24/29 cases), whenmost social interactions occur, which supports the social subordinationhypothesis. Lynxes left the natal range at an earlier age fromthe local population with higher density, which supports intraspecificcompetition for resources as a cause for dispersal. However,prey seems not to be the limiting resource because dispersalmostly started during the season of increasing prey density,and individual probability of dispersal increases with preydensity in the natal area. Dispersers suffered higher mortalitythan nondispersing lynxes of the same age, due mostly to anthropogeniccauses. About half of the dispersers successfully settled ina territory at distances that differed between areas of origin,but not between sexes. Distances reached, shorter than thosereported for other similar-size solitary felids, are limitedby anthropogenic barriers that prevent connection with closermetapopulations.     

Ectoparasites (sucking lice,fleas and ticks) of small mammals in southeastern Kenya

During 1998–2000, at least 14 species (n = 309) of small mammals were live‐trapped and examined for ectoparasites in moist forests of the Taita and Shimba Hills and drier savannah habitats of Nguruman, southeastern Kenya. Ectoparasites were recorded from 11 species of mammals. Five species of sucking lice [Hoplopleura inexpectans Johnson, H. intermedia Kellogg & Ferris, Polyplax reclinata (Nitzsch), P. waterstoni Bedford and Schizophthirus graphiuri Ferris], six species of fleas (Ctenophthalmus leptodactylous Hubbard, Dinopsyllus grypurus Jordan & Rothschild, D. lypusus Jordan & Rothschild, Hypsophthalmus campestris Jordan & Rothschild, Listropsylla basilewskyi Smit and Xiphiopsylla lippa Jordan) and at least six species of ticks (Amblyomma sp., Haemaphysalis sp., Ixodes sp., I. alluaudi Neumann, I. cumulatimpunctatus Schulze, I. muniensis Arthur & Burrow and Rhipicephalus sp.) were recorded from these hosts. Four of the five species of sucking lice were host specific whereas P. reclinata was recorded from two different species of white‐toothed shrews, Crocidura spp. Although fleas and ticks were less host specific, C. leptodactylous, D. grypurus and I. cumulatimpunctatus were only recorded from the murid rodent Praomys delectorum (Thomas), Amblyomma sp. was only recorded from the nesomyid rodent Beamys hindei Thomas, Rhipicephalus sp. was only recorded from the murid Lemniscomys striatus (L.) and I. muniensis was only recorded from the dormouse Graphiurus microtis (Noack). More species of ectoparasites and significantly greater infestation prevalences were recorded from small mammals in moist habitats compared with those from the savannah habitat. At least one of the fleas recorded, D. lypusus, is a known vector of Yersinia pestis Lehmann & Neumann, the causative agent of plague, which is present in the region.     

Faecal genetic analysis to determine the presence and distribution of elusive carnivores: design and feasibility for the Iberian lynx

Noninvasive methods using genetic markers have been suggested as ways to overcome difficulties associated with documenting the presence of elusive species. We present and assess a novel, reliable and effective molecular genetic technique for the unequivocal genetic identification of faeces from the endangered Iberian lynx (Lynx pardinus). From mitochondrial DNA (mtDNA) cytochrome b and D-loop region sequences, we designed four species-specific primers (for products 130-161 bp long) that were considered to be likely to amplify degraded DNA. We compared two DNA extraction methods, various DNA amplification conditions and the robustness and specificity of the primer pairs with 87 lynx samples from 5 potentially different lynx populations and with 328 samples of other carnivore species. The utility of the identification technique was tested with faeces of different ages, with faeces from controlled field experiments, and with faeces collected from locales with possible lynx populations from throughout the state of Andalusia, Spain (8052 km2). Faecal mtDNA extraction was more efficient using PBS wash of the faeces instead of a faeces homogenate. Our assay increased from 92.6 to 99% efficiency with a second amplification and a reduction in template concentration to overcome polymerase chain reaction (PCR) inhibition. Our assay never produced false positives, and correctly identified all lynx faeces. Of 252 faeces samples of unknown species collected throughout Andalusia, 26.6% (from three different areas) were classified as Iberian lynx, 1.4% showed evidence of PCR inhibition and 1.2% were of uncertain origin. This method has proven to be a reliable technique that can be incorporated into large-scale surveys of Iberian lynx populations and exemplifies an approach that can easily be extended to other species.     

Ectoparasites of commensal rodents in Sulawesi Utara, Indonesia, with notes on species of medical importance

Ectoparasite records are presented for four species of commensal murid rodents (Rattus rattus palelae Miller & Hollister, R. argentiventer (Robinson & Kloss), R. exulans (Peale) and Mus musculus castaneus Waterhouse) in Sulawesi Utara, with particular reference to the potential for these arthropods to bite and transmit pathogens to humans. The flea, Xenopsylla cheopis (Rothschild), was most common on R.r. palelae and is capable of transmitting plague and other pathogens to humans although no current foci for these diseases are known in Sulawesi. Hoplopleura pacifica Ewing and Polyplax spinulosa (Burmeister) sucking lice parasitized all three Rattus species although H. pacifica was mainly associated with R. exulans and P. spinulosa with R.r. palelae. These lice do not bite humans but may be intramurid vectors of murine typhus and other zoonoses. The mites Laelaps echidnina Berlese and L. nuttalli Hirst were both collected; the latter was recorded from all four murid species, mainly R. exulans. The mite Ornithonyssus bacoti Hirst was rare. Only one chigger mite, Walchiella oudemansi (Walch), was retrieved from murids (from R. exulans) and a single Leptotrombidium deliense (Walch) chigger was taken from a human subject. Although L. deliense is a significant vector of scrub typhus, a disease known from Sulawesi, the L. deliense-R. argentiventer relationship frequently noted in the ecology of this rickettsial disease, was not evident in this survey. Other ectoparasites collected from murids were the ticks, Ixodes granulatus Supino (first record for Sulawesi), Haemaphysalis sp. and Dermacentor sp., the mites Myocoptes musculinus (Koch) and Listrophoroides cucullatus (Trouessart), acarids and a uropodid.(ABSTRACT TRUNCATED AT 250 WORDS)     

Ectoparasites of the Pallas squirrel, Callosciurus erythraeus, introduced to Japan

The squirrel Callosciurus erythraeus (Pallas) (Rodentia: Sciuridae) was intentionally introduced to Japan in 1935 and has become established throughout much of the country. Although they live mainly in forests, Pallas squirrels come into gardens and are frequently fed by people or kept as pets, so their ectoparasites could be of potential medical as well as veterinary importance. During 2001-2003 we conducted the first ectoparasite survey of Pallas squirrels in Japan. From 105 C. erythraeus captured in Kamakura District of Kanagawa Prefecture on Honshu Island, three types of ectoparasite were found: 52 specimens of the sucking louse Neohaematopinus callosciuri Johnson (Anoplura: Haematopinidae), 26 fleas Ceratophyllus (Monopsyllus) anisus Rothschild (Siphonaptera: Ceratophyllidae) and four nymphs of the tick Haemaphysalis flava Neumann (Acari: Ixodidae) on 22, 13 and one squirrels, respectively. Evidently in Japan C. erythraeus carries relatively few ectoparasite species; this may be a contributory factor to their invasive success. Further investigations are needed to assess risks of zoonotic transmission of plague or murine typhus by C. anisus, of louse-borne typhus by N. callosciuri and of tularaemia and especially Japanese spotted fever (Rickettsia japonica) by H. flava.     

Abiotic factors influencing the ecology of wild rabbit fleas in north-eastern Spain

During 1992, the population dynamics of rabbit fleas were compared at two sites in north-eastern Spain. The sites differed mainly in terms of annual rainfall and soil type. All flea species showed seasonal cycles of abundance, although peaks in numbers occurred at different times, reflecting their specific adaptations for coping with climatic variables. Adult Spilopsyllus cuniculi (Dale) (Siphonaptera: Pulicidae) were found largely parasitizing rabbits in spring and adult Caenopsylla laptevi (Beaucournu etal.) (Siphonaptera: Ceratophyllidae) in the autumn. In contrast, monthly flea indices of Xenopsylla cunicularis (Smit) (Siphonaptera: Pulicidae) and Echidnophaga iberica (Ribeiro et al.) (Siphonaptera: Pulicidae) peaked in summer. Spilopsyllus cuniculi was present at both sites, but was less common on the drier site, where monthly mean temperature and annual rainfall approached the flea's physiological limits. By contrast, E. iberica, X. cunicularis and C. laptevi, known to be better adapted for dryness, showed the opposite patterns of abundance. Nevertheless, even these arid-adapted species took advantage of the milder and wetter spring (X. cunicularis and E. iberica) or autumn (C. laptevi) for breeding and larval development. Although environmental temperature, rainfall and soil texture will influence the microclimate of the burrows where the flea larvae develop, burrow humidity seems to be more dependent on soil characteristics and past rainfall rather than the humidity of the external air.     

Cryobanking the genetic diversity in the critically endangered Iberian lynx (Lynx pardinus) from skin biopsies. Investigating the cryopreservation and culture ability of highly valuable explants and cells

Cryobanking skin samples permit preserving a maximum of genetic representation from the population biodiversity. This is a relevant aspect for threatened species, potentially menaced by an epizooty and from which it is difficult to obtain gametes. As a first step for properly cryobanking skin samples of a given species, the optimal conditions of culture and freezing have to be studied by covering a broad range of possibilities. This paper presents, for the first time, a systematic study of such conditions for the Iberian lynx (Lynx pardinus). To that end, we have analyzed twenty different culture conditions and fifteen different freezing solutions for skin explants, as well as three freezing solutions for isolated cells derived from them. The culture conditions included both two different culture strategies and several combinations of nutritional supplements and mitotic agents. For the freezing solutions, we have considered different concentrations of the permeating cryoprotectant dimethyl sulfoxide (Me₂SO) either alone (5%, 7.5%, 10%, 12.5% and 15% v/v for explants, 10% for isolated cells) or along with the non-permeating cryoprotectant sucrose (0.1 or 0.2 M). Our results have been analyzed through several quantitative parameters and show that only thawed explants cryopreserved in Me₂SO (10%) either alone or with sucrose (0.2 M) presented similar properties to those in optimal fresh cultures. In addition, for these freezing conditions, isolated thawed cells also presented high survival rates (90%) and percentages of cellular functionality (85%). These results, focussed on the most endangered felid in the world, could be also useful for other threatened/endangered species.     

Species-specific mitochondrial DNA markers for identification of non-invasive samples from sympatric carnivores in the Iberian Peninsula

Genetic species identification of non-invasively collected samples has become an important tool in ecological research, management and conservation and wildlife forensics. This is especially true for carnivores, due to their elusive nature, and is crucial when several ecologically and phylogenetically close species, with similar faeces, hairs, bones and/or pelts, occur in sympatry. This is the case of the Iberian Peninsula, a region with a carnivore community of 16 species—about two-thirds of the European carnivore fauna. Here we present a simple, efficient and reliable PCR-based protocol, using a novel set of species-specific primers, for the unambiguous identification to species of non-invasively collected samples or forensic materials from Iberian carnivores. For each species, from the consensus of all cytochrome b haplotypes, found here and previously reported, we designed species-specific primer pairs for short fragments, the most likely to persist in low-quantity and degraded DNA samples. The predicted specificity of each primer pair was assessed through PCR of positive DNA extracts from the carnivore species, from an exhaustive array of potential prey and from humans. The robustness of PCR amplification for non-invasively sampled DNA was tested with scat samples. The primers did not produce false positives and correctly identified all carnivore samples to the species level. In comparison with sequencing and PCR-RFLP assays, our method is, respectively, cost- and time-effective, and is especially suited for monitoring surveys targeting multiple populations/species. It also introduces an approach that works for a whole community of carnivores living sympatrically over a large geographic area.     

Revision and new data of the ammonite family Pseudotissotiidae in the Iberian Trough, Spain

The ammonites assigned to the family Pseudotissotiidae of the Wiedmann (Tübingen, Germany) and Goy, Carretero and Meléndez (Madrid, Spain) collections obtained from the Iberian Trough have been revised. New, mainly lower Turonian, specimens of the species Pseudotissotia sp., Choffaticeras (Choffaticeras) quaasi (Peron, 1904), C. (C.) pavillieri (Pervinquière, 1907), C. (C.) segne (Solger, 1903), C. (C.) sinaiticum (Douvillé, 1928), C. (Leoniceras) luciae (Pervinquière, 1907), C. (L.) barjonai (Choffat, 1898), Wrightoceras llarenai (Karrenberg, 1935), W. munieri (Pervinquière, 1907), W. submunieri Wiedmann, 1975b, and Donenriquoceras forbesiceratiforme Wiedmann, 1960, have been also presented. In addition, we designate and refigure the lectotype of Pseudotissotia (Leoniceras) alaouitense Basse, 1937, and refigure the lectotype of P. barjonai Choffat, 1898. Studies on the morphologies and the geographical and temporal distributions of all of these species have led to the identification of several phylogenetic relationships between them, and to distinguish four main phases in the evolution of the family, characterized by the successive dominance of Choffaticeras (Choffaticeras), Choffaticeras (Leoniceras), Donenriquoceras and Wrightoceras with Donenriquoceras.     

Episodic development of Cambrian eocrinoid-sponge meadows in the Iberian Chains (NE Spain)

Summary The Lower-Middle Cambrian transition in the Iberian Chains (NE Spain) records a transgressive deposition from peritidal to open-shelf environments. This overall trangressive trend took place across an interval of syndepositional tectonic processes. which have been illustrated during late Bilbilian to Leonian times. The scarcity of building organisms in this areas seems to be directly related to a rhythmic influence of fine terrigenous decantation, which inhibited and interrupted the carbonate production. Only in the more proximal and quiet environments, the episodic establishment of stromatolitic boundstones is evidence. Eocrinoid-sponge meadows were developed in shallow subtidal environments, and were completed by the presence of trilobites, inarticulate brachiopods, hyoliths and Chancelloriid sclerites. The biggest accumulation rates associated to these complex meadow communities seem to be reflected by differential synsedimentary faulting processes.