House sparrow uropygial gland secretions do not attract ornithophilic nor mammophilic mosquitoes.

Mosquito feeding preferences determine host-vector contact rates and represent a key factor in the transmission of vector-borne pathogens. The semiochemical compounds of which vertebrate odours are composed probably play a role in mosquito host choice. Birds spread secretions from uropygial gland over their feathers to protect their plumage, comprising behaviour that may in turn affect odour profiles. Although uropygial secretions are expected to modify the attractiveness of birds to mosquitoes, contradictory findings have been reported. Mosquito species differ in their feeding preferences, with some species feeding mainly on birds (ornithophilic species) and others on mammals (mammophilic species). Consequently, it is possible that ornithophilic and mammophilic species differ in their response to uropygial gland secretions. Using a dual-choice olfactometer, the present study tests this hypothesis by comparing the behavioural response to uropygial gland secretions from juvenile male and female house sparrows (Passer domesticus) in the ornithophilic Culex pipiens and the mammophilic Aedes (Ochlerotatus) caspius mosquitoes. No differences were found in the response of either mosquito species to the uropygial gland secretions. Therefore, the preference of ornithophilic mosquitoes for avian hosts is apparently not explained by a greater attraction of mosquitoes to the uropygial gland secretion odour when presented in combination with a CO2 -enriched airflow.

A spatial ecology study in a high-diversity host community to understand blood-feeding behaviour in Phlebotomus sandfly vectors of Leishmania.

Molecular studies indicate that Phlebotomine sandflies (Diptera: Psychodidae) blood feed on many vertebrate species, of which only a few are proven parasite reservoirs. Investigating sandfly vector feeding preferences is therefore important and requires taking into account the availability and accessibility of host species. In terms of the latter, it is necessary to consider the metabolic cost to the insect of reaching the host and moving on to a suitable breeding site. The present study used statistical modelling to compare the feeding patterns of Phlebotomus perniciosus (n = 150), Phlebotomus papatasi (n = 35) and Phlebotomus ariasi (n = 7) on each of an average of 30 host species in a wildlife park in Murcia, Spain. Sandfly feeding movement costs were estimated as a function of the distance and altitude gradients saved by the insect, assuming that they displayed 'site fidelity'. Most (87%) engorged females were caught <100 m from the host on which they had fed. Although the percentage of bloodmeals was highest on fallow deer (Dama dama) (30%) and red deer (Cervus elaphus) (26%), the predicted feeding probability after considering movement cost was highest for red deer and common eland (Taurotragus oryx), and positively associated with host census. These results suggest that, under similar circumstances, sandflies prefer to feed on some host species more than on others.

Endemic species may have complex histories: within-refugium phylogeography of an endangered Iberian vole.

Glacial refugia protected and promoted biodiversity during the Pleistocene, not only at a broader scale, but also for many endemics that contracted and expanded their ranges within refugial areas. Understanding the evolutionary history of refugial endemics is especially important in the case of endangered species to recognize the origins of their genetic structure and thus produce better informed conservation practices. The Iberian Peninsula is an important European glacial refugium, rich in endemics of conservation concern, including small mammals, such as the Cabrera vole (Microtus cabrerae). This near-threatened rodent is characterized by an unusual suite of genetic, life history and ecological traits, being restricted to isolated geographic nuclei in fast-disappearing Mediterranean subhumid herbaceous habitats. To reconstruct the evolutionary history of the Cabrera vole, we studied sequence variation at mitochondrial, autosomal and sex-linked loci, using invasive and noninvasive samples. Despite low overall mitochondrial and nuclear nucleotide diversities, we observed two main well-supported mitochondrial lineages, west and east. Phylogeographic modelling in the context of the Cabrera vole's detailed fossil record supports a demographic scenario of isolation of two populations during the Last Glacial Maximum from a single focus in the southern part of the Iberian Peninsula. In addition, our data suggest subsequent divergence within the east, and secondary contact and introgression of the expanding western population, during the late Holocene. This work emphasizes that refugial endemics may have a phylogeographic history as rich as that of more widespread species, and conservation of such endemics includes the preservation of that genetic legacy.

First molecular identification of the vertebrate hosts of Culicoides imicola in Europe and a review of its blood-feeding patterns worldwide: implications for the transmission of bluetongue disease and African horse sickness.

Culicoides (Diptera: Ceratopogonidae) are vectors of pathogens that affect wildlife, livestock and, occasionally, humans. Culicoides imicola (Kieffer, 1913) is considered to be the main vector of the pathogens that cause bluetongue disease (BT) and African horse sickness (AHS) in southern Europe. The study of blood-feeding patterns in Culicoides is an essential step towards understanding the epidemiology of these pathogens. Molecular tools that increase the accuracy and sensitivity of traditional methods have been developed to identify the hosts of potential insect vectors. However, to the present group's knowledge, molecular studies that identify the hosts of C. imicola in Europe are lacking. The present study genetically characterizes the barcoding region of C. imicola trapped on farms in southern Spain and identifies its vertebrate hosts in the area. The report also reviews available information on the blood-feeding patterns of C. imicola worldwide. Culicoides imicola from Spain feed on blood of six mammals that include species known to be hosts of the BT and AHS viruses. This study provides evidence of the importance of livestock as sources of bloodmeals for C. imicola and the relevance of this species in the transmission of BT and AHS viruses in Europe.

West Nile virus-neutralizing antibodies in wild birds from southern Spain.

West Nile virus (WNV) is an emerging vector-borne arbovirus with a zoonotic life-cycle whose main reservoir hosts are birds. In humans and horses, WNV infections rarely result in clinical disease but on occasions - depending on factors such as climatic conditions, insect communities and background immunity levels in local populations - they can lead to outbreaks that threaten public and animal health. We tested for the presence of WNV antibodies in 149 birds belonging to 32 different species. Samples were first tested using a bird-specific ELISA kit and then both positive and doubtful results were confirmed by neutralization tests using WNV and Usutu virus. WNV antibodies were confirmed in a resident Sylvia melanocephala juvenile, supporting the idea of local transmission of WNV in southern Spain in 2013. In addition, the serum from an adult blackbird (Turdus merula) showed neutralization of both WNV and Usutu virus. We discuss our results in light of the occurrence of WNV on horse farms in southern Spain in 2013.

First reported case of fatal tuberculosis in a wild African elephant with past human-wildlife contact.

Tuberculosis is emerging/re-emerging in captive elephant populations, where it causes morbidity and deaths, although no case of TB in wild African elephants has been reported. In this paper we report the first case of fatal TB in an African elephant in the wild. The infection with Mycobacterium tuberculosis was confirmed by post-mortem and histological examinations of a female sub-adult elephant aged >12 years that died in Tsavo East National Park, Kenya, while under treatment. This case is unique in that during its lifetime the elephant had contact with both humans and wild elephants. The source of the infection was unclear because the elephant could have acquired the infection in the orphanage or in the wild. However, our results show that wild elephants can maintain human TB in the wild and that the infection can be fatal.

Molecular phylogeny and systematics of the highly polymorphic Rumex bucephalophorus complex (Polygonaceae).

Rumex bucephalophorus is a very polymorphic species that has been subjected to various taxonomic studies in which diverse infraspecific taxa have been recognised on the basis of diaspore traits. In this study we used molecular markers (ITS and AFLP) to explore this remarkable diversity, to test previous hypotheses of classification, and attempt to explain biogeographic patterns. Results show that R. bucephalophorus forms a monophyletic group in which diversification began around 4.2 Mya, at the end of Messinian Salinity Crisis. The two molecular markers clearly show a deep divergence separating subsp. bucephalophorus from all other subspecific taxa, among which subsp. canariensis also constitutes a separate and well distinguishable unit. In contrast, subspecies hispanicus and subsp. gallicus constitute a monophyletic group in which three subgroups can be recognised: subsp. hispanicus, subsp. gallicus var. gallicus and subsp. gallicus var. subaegeus. However, these three subgroups are not clearly distinguished genetically or morphologically, so that in formal classification it would be preferable to treat them at the varietal level.

Is ITS-2 rDNA suitable marker for genetic characterization of Sarcoptes mites from different wild animals in different geographic areas?

The present study examined the relationship among individual Sarcoptes scabiei mites from 13 wild mammalian populations belonging to nine species in four European countries using the second internal transcribed spacer (ITS-2) of nuclear ribosomal DNA (rDNA) as genetic marker. The ITS-2 plus primer flanking 5.8S and 28S rDNA (ITS-2+) was amplified from individual mites by polymerase chain reaction (PCR) and the amplicons were sequenced directly. A total of 148 ITS-2+ sequences of 404bp in length were obtained and 67 variable sites were identified (16.59%). UPGMA analyses did not show any geographical or host-specific clustering, and a similar outcome was obtained using population pairwise Fst statistics. These results demonstrated that ITS-2 rDNA does not appear to be suitable for examining genetic diversity among mite populations.

Attract them anyway: benefits of large, showy flowers in a highly autogamous, carnivorous plant species.

Reproductive biology of carnivorous plants has largely been studied on species that rely on insects as pollinators and prey, creating potential conflicts. Autogamous pollination, although present in some carnivorous species, has received less attention. In angiosperms, autogamous self-fertilization is expected to lead to a reduction in flower size, thereby reducing resource allocation to structures that attract pollinators. A notable exception is the carnivorous pyrophyteDrosophyllum lusitanicum(Drosophyllaceae), which has been described as an autogamous selfing species but produces large, yellow flowers. Using a flower removal and a pollination experiment, we assessed, respectively, whether large flowers in this species may serve as an attracting device to prey insects or whether previously reported high selfing rates for this species in peripheral populations may be lower in more central, less isolated populations. We found no differences between flower-removed plants and intact, flowering plants in numbers of prey insects trapped. We also found no indication of reduced potential for autogamous reproduction, in terms of either seed set or seed size. However, our results showed significant increases in seed set of bagged, hand-pollinated flowers and unbagged flowers exposed to insect visitation compared with bagged, non-manipulated flowers that could only self-pollinate autonomously. Considering that the key life-history strategy of this pyrophytic species is to maintain a viable seed bank, any increase in seed set through insect pollinator activity would increase plant fitness. This in turn would explain the maintenance of large, conspicuous flowers in a highly autogamous, carnivorous plant.

Estimation of gene diversity at the b locus of the constant region of the immunoglobulin light chain in natural populations of European rabbit (Oryctolagus cuniculus) in Portugal, Andalusia and on the Azorean Islands.

The minimal gene diversity at a locus of the antibody constant region, as estimated in natural populations of rabbit, revealed levels of heterozygosity similar to those reported for the major histocompatibility complex in human and murine populations. Sera of 416 wild rabbits were collected on the Iberian peninsula and on three islands of the Azorean archipelago and analyzed for the occurrence of the serological markers of the b locus of the immunoglobulin light chain. All four serotypes present in domestic rabbits were found in Portugal. They represented less than 50% of the gene pool. In Andalusia this was less than 15% and on the Azorean islands less than 10%. The pronounced and systematic hierarchy in allele frequencies, previously found in populations from the more recent distribution area of the species, was not observed. On the peninsula, the frequencies of the "domestic" alleles were similar, averaging 10%. The Portuguese sample revealed a total heterozygosity of at least 87%. This high value was supported by at least 11 serologically different alleles, none of them occurring at frequencies above 20%. These data are in agreement with an Iberian origin of the European rabbit and strongly suggest the coalescence of b locus allelic lines drawn from Iberian and western populations. The role of balancing selection in the evolution of the b locus polymorphism was further emphasized.

Genotypic identification of an undescribed spotted fever group rickettsia in Ixodes ricinus from southwestern Spain.

An undescribed rickettsia was directly analyzed with specific rickettsial molecular biology tools on Ixodes ricinus L. collected in different localities of the province of Cadiz (southwestern Spain). On the basis of the results of the citrate synthase (glta) gene, 190 kD-outer membrane protein (rOmpA) gene, and 16S ribosomal RNA (16S rRNA) gene partial sequence data, it was found that this rickettsia is sufficiently genetically distinct from other Rickettsia to be considered a distinct taxonomic entity. The isolation and culture of this organism, as well as comparative antigenic analysis, are required to ensure its conclusive taxonomic placement among spotted fever rickettsiae. The epidemiologic role of this new rickettsial agent and its possible pathogenicity to wild and domestic animals or humans is still unknown and needs to be investigated.

Pharyngeal bot flies (Oestridae) from sympatric wild cervids in southern Spain.

The prevalence of 2 species of pharyngeal bot flies parasitizing sympatric wild ungulates from southern Spain is given. Pharyngomyia picta was found in all red deer and fallow deer whereas Cephenemyia auribarbis was found in 59% of fallow deer and 81% of red deer examined. The mean number of P. picta larvae was 3-6 times more than that of C. auribarbis larvae. These data suggest that red deer are more heavily parasitized than fallow deer and that they are predominantly infested with P. picta larvae.

Prevalence and seasonality of Oestrus caucasicus Grunin, 1948 (Diptera: Oestridae) parasitizing the Spanish ibex, Capra pyrenaica (Mammalia: Artiodactyla).

Prevalence and intensity of parasitism by oestrid larvae in the Spanish ibex Capra pyrenaica were determined for a period of 2.5 yr in Sierra Nevada Natural Park, southern Spain. The most relevant result of this study was to find Oestrus caucasicus parasitizing this ungulate in a geographical location far from its previously known distribution, e.g., the Caucasus mountain range and central Asia. Larvae were found within 74% of the heads examined (n = 180). Prevalence was higher in females and in older animals. Lowest prevalence occurred during the summer months. The mean intensity (+/- SD) was 25.4 +/- 27.3 larvae and was similar in both host sexes. The number of larvae increased with host age and reached the highest intensities in winter, during December and January.

Oestrid myiasis in European mouflon from Spain.

From February 1992 to March 1997, 245 European mouflon (Ovis orientalis musimon) from Sierras de Cazorla, Segura y Las Villas Natural Park (southern Spain) were surveyed for oestrid larvae in order to estimate prevalence and mean intensity of parasitism by Oestrus ovis. Over 46 percent of the animals surveyed were infected, with a mean intensity of 9.6 larvae/host parasitized. No significant differences in prevalence rates between host sexes were observed, but older mouflons were infected with more larvae than younger ones.

Oestrosis in red deer from Spain.

A survey of naso-pharyngeal myiasis affecting red deer (Cervus elaphus) in southern Spain was conducted. The parasites involved were the larvae of Pharyngomyia picta and Cephenemyia auribarbis (Diptera:Oestridae), which coexist sympatrically within this host. Males and older animals had higher prevalences and intensities of fly larvae. Differences in behaviour and habitat use by male and female deer, and the increase of head size in older males are possibly responsible for this. There were low densities of C. auribarbis while P. picta was the species most frequently observed, although both oestrids were located in the same host cavities. The earlier larviposition by C. auribarbis, and its faster larval development may reflect asynchronous life-cycles of both oestrids; this may decrease inter-specific competition between these sympatric species.

Hematologic parameters of the Spanish ibex (Capra pyrenaica).

Thirteen hematologic parameters were measured in 52 Spanish ibexes (Capra pyrenaica) from Sierra Nevada Natural Park, southern Spain. The animals were captured, maintained in captivity, and physically restrained for blood collection. We compared hematologic values for healthy ibexes with animals infected with sarcoptic mites and animals treated for scabies and recovered. Ibexes with scabies showed a decreased number of erythrocytes, a higher mean corpuscular volume and mean corpuscular hemoglobin, and increased numbers of band neutrophils. The number of lymphocytes was significantly higher in animals that had recovered from scabies as compared with infested ones. The only difference observed between healthy and recovered animals was in the number of monocytes, which remained significantly higher in animals that had recovered from scabies. No hemoparasites were found in this study.

Andromonoecy in an Old World Papilionoid legume, Erophaca baetica.

Andromonoecy (i.e. the occurrence on individual plants of hermaphroditic and male flowers) is a rare sexual system among the angiosperms, regarded by some authors as a transitional stage from hermaphroditism to monoecy. Having discovered the occurrence of andromonoecy in Erophaca baetica (a Mediterranean shrubby legume with two subspecies), a novelty for Old World papilionoid legumes, we investigated the morpho-functional correlates and the geographical distribution of this phenomenon in the species. The relative frequencies of hermaphrodite and male flowers were determined in two field and 111 herbarium populations. Biomass allocation within flowers, pollen production and viability, pollen tube growth, nectar production and the temporal pattern of male flower production were also studied in two nearby southern Spanish populations. Virtually all of the studied populations were andromonoecious. Male flowers tended to appear at apical positions within the inflorescence, and became more abundant by the end of the flowering season. Male flowers were externally similar to hermaphroditic flowers (although with less biomass and smaller parts) and released equivalent amounts of pollen and nectar; however, their pollen germinated significantly better. Erophaca is the first example of an andromonecious Papilionoid in the Old World. Since the main difference among floral morphs in this species is functional (i.e. pollen germination rate) rather than morphological, andromonoecy is not readily noticeable, and very careful inspection may be required to reveal it. The potential effect of andromonoecy in enhancing outcrossing rate in this species is discussed.

The opportunistic Sarcoptes scabiei: a new episode from giraffe in the drought-suffering Kenya.

The ubiquitous Sarcoptes mite is unexplainable emerging and re-emerging parasite, threatening biodiversity and human health. When a new outbreak occurs, it is not clear if it is a genuine emergence resulting from a new incidence or apparent emergence resulting from increased detection. In this paper we report, for the first time to our knowledge, an outbreak of sarcoptic mange in giraffes in the wild. Three decaying carcasses and five free-ranging subadult reticulated giraffes were observed to have mange-like lesions in the drought-suffering Wajir Region in North Eastern Kenya, while apparently all sympatric wild and domestic animals were mange-free. Affected giraffes were captured and successfully treated. The possible relations between this outbreak and annual seasons, animal age-classes and sex, and spatial distribution are discussed.

Sarcoptes mite from collection to DNA extraction: the lost realm of the neglected parasite.

Sarcoptes mite from collection to DNA extraction forms the cornerstone for studies on Sarcoptes scabiei. Whilst the new science era took a shy leap into the different facets of mite studies, the cornerstone was almost entirely neglected. Mite collection, cleaning, storage and DNA extraction were, basically, humble attempts to extrapolate, adapt, modify or 'pirate' those existing methods to the peculiarities of Sarcoptes research. These aspects usually constituted few lines, bashfully mentioned, in the materials and methods section of some papers, which arose in unique problems concerning cost-effectiveness, time profitability, safety and even worse, the credibility of the results, creating contradictory conclusions in some cases. This 'noisy' situation encouraged us to collect, classify and review, for the first time to our knowledge, some aspects relating to studies on Sarcoptes mite from collection to DNA extraction, which will be useful for further studies on Sarcoptes, and have implications for the effective control of the diseases Sarcoptes mite causes. Further studies are needed, especially to compare the profitability, safety, sensibility and specificity of the different methods of this neglected realm of the ubiquitous ectoparasite.

Bronchopulmonary nematode infection of Capra pyrenaica in the Sierra Nevada massif, Spain.

The present investigation examined the prevalence and abundance of bronchopulmonary nematodes in 213 randomly hunted Iberian ibexes (Capra pyrenaica) (87 females and 126 males) in the Sierra Nevada mountain range in Spain between 2003 and 2006. Post mortem examination revealed an overall prevalence of 72% for adult nematodes (Cystocaulus ocreatus 44%, Muellerius capillaris 44%, Protostrongylus sp. 40%, and Dictyocaulus filaria 4%). The abundances were 13.45+/-3.97, 5.18+/-2.49, 6.36+/-2.16, and 2.27+/-0.46, respectively. Protostrongylid adults showed similar infection rates, which were statistically different from that of D. filaria. 20% of the examined Iberian ibexes were infected by three protostrongylid nematodes species, 24% of C. pyrenaica were affected by two protostrongylid species, while infestations with only one protostrongylid species were detected in 20% of the examined animals. The overall prevalence of larvae nematodes in the examined animals was 100%, and the overall abundance (number of the first stage larvae per gram) was 86.45+/-20.63. There was a high correlation between the two sets of data (adults and larvae). Results of the present investigation provided foundation for the effective control of bronchopulmonary nematode infection in Iberian ibex.