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Raccoons are an invasive species continuously spreading throughout Europe with Germany as the core area. Here, raccoons mostly live in rural regions, but some urban populations are already established (city of Kassel) or starting to build up (Berlin). The objective of this study was to investigate Sarcoptes infections in racoons in these two urban areas and try to identify the putative origin of the parasite. Parasite morphology, gross and histopathology examinations of diseased skin tissue were consistent with Sarcoptes scabiei infection. Moreover, using 9 microsatellite markers we genotyped individual mites from five raccoons and compared them with fox-, wild boar- and Northern chamois-derived Sarcoptes mites, originating from Italy and Switzerland. The raccoon-derived mites clustered together with the fox samples and were clearly differentiated from those of the wild boar and chamois suggesting a fox origin for the raccoon mange infection. These results are evidence of cross-transmission of S. scabiei among wild carnivores. Although our results cannot elucidate whether raccoons became infected by frequent interaction with endemically or epidemically infected foxes or if these cases were the result of occasional contacts among these animal species. Nevertheless, they show that pathogens can be shared among urban populations of native and invasive carnivores

Background: Human-elephant conflict (HEC) is a recurring problem that appears wherever the range of elephants and humans overlap. Different methods including the use of electric fences are used worldwide to mitigate this conflict. Nonetheless, elephants learn quickly that their tusks do not conduct electricity and use them to break down fences (fencebreakers). Methodology/Principal Findings: In Lewa Wildlife Conservancy, Kenya, destructive elephants (Loxodonta africana) were monitored between 2010 and 2013. The fence-breaking rate reached four incidents (fence-breaking) per elephant per 100 days. Ten bull males and 57 females were identified as fence-breakers. The bulls were involved in 85.07% and the females in 14.93% of incidents. The Kenya Wildlife Service approved detusking (partial cutting of tusks) in four of the 10 fence-breakers as a way of preventing them from breaking down fences, thereby mitigating HEC in the Conservancy. The result of the detusking was a drastic six-fold reduction in damage to fences (range: 1.67 to 14.5 times less fence-breaking) by the four worst fence-breaker elephants, because with trimmed tusks elephants lack the tools to break down fences. Detusking could not totally eliminate fence destruction because, despite lacking their tools, elephants can still destroy fences using their heads, bodies and trunks, albeit less effectively. On the other hand, apart from inherent aesthetic considerations, the detusking of elephants may have certain negative effects on factors such as elephants' social hierarchies, breeding, mate selection and their access to essential minerals and food. Conclusions: Elephant detusking seems to be effective in drastically reducing fence-breaking incidents, nonetheless its negative effects on behaviour, access to food and its aesthetical consequences still need to be further studied and investigated. © 2014 Mutinda et al.

[Background] The parasitic nematodes Ascaris lumbricoides and A. suum are of great public health and economic significance, and the two taxa were proposed to represent a single species. miRNAs are known with functions of gene regulations at post-transcriptional level., [Results] We herein compared the miRNA profiles of A. lumbricoides and A. suum female adults by Solexa deep sequencing combined with bioinformatics analysis and stem-loop real-time PCR. Using the A. suum genome as the reference genome, we obtained 171 and 494 miRNA candidates from A. lumbricoides and A. suum, respectively. Among which, 74 miRNAs were shared between the two taxa, 97 and 420 miRNAs were A. lumbricoides and A. suum specific. Target and function prediction revealed a significant set of targets which are related to ovarian message protein, vitellogenin and chondroitin proteoglycan of the two nematodes. Enrichment analysis revealed that the percentages of most predicted functions of the miRNA targets were similar, with some taxon specific or taxon enhanced functions, such as different target numbers, specific functions (NADH dehydrogenase and electron carrier functions), etc., [Conclusions] This study characterized comparatively the miRNAs of adult A. lumbricoides and A. suum, and the findings provide additional evidence that A. lumbricoides and A. suum represent a single species. Due to the fast evolution nature of miRNAs and the different parasitic living conditions of humans and pigs, the phenomenon above might indicate a fast evolution of miRNAs of Ascaris in humans and pigs.

The discipline of epidemiology is the study of the patterns, causes and effects of health and disease conditions in defined anima populations. It is the key to evidence-based medicine, which is one of the cornerstones of public health. One of the important facets of epidemiology is disease-navigating webs (disease-NW) through which zoonotic and multi-host parasites in general move from one host to another. Epidemiology in this context includes (i) classical epidemiological approaches based on the statistical analysis of disease prevalence and distribution and, more recently, (ii) genetic approaches with approximations of disease-agent population genetics. Both approaches, classical epidemiology and population genetics, are useful for studying disease-NW. However, both have strengths and weaknesses when applied separately, which, unfortunately, is too often current practice. In this paper, we use Sarcoptes scabiei mite epidemiology as a case study to show how important an integrated approach can be in understanding disease-NW and subsequent disease control.

Background Wildlife radio tracking has gained popularity during the recent past. Ecologists and conservationists use radio-collars for different purposes: animal movement monitoring, home range, productivity, population estimation, behaviour, habitat use, survival, and predator-prey interaction, among others. The aim of our present study is to highlight the application of radio-collars for wildlife diseases monitoring. The spread of wildlife diseases and the efficacy of management actions for controlling them propose serious challenges for ecologists and conservationists, since it is difficult to re-capture (or simply observe) the same animal in pre-determined temporal interval, but such difficulty is overcome by the use of gps-gsm radio collars. Methods In the present study we report, for the first time to our knowledge, the use of radio-collars in the monitoring of Iberian ibex affected by Sarcoptes scabiei in Sierra Nevada mountain range, Spain. Twenty-five moderate or slightly mangy animals were radio-collared between 2006 and 2013. Results The radio-collars allowed us to confirm the presence of resistance to S. scabiei within Iberian ibex population. Twenty (80%) of the collared animals recovered totally from mange, while the disease progressed in the other five Iberian ibex (20% of the collared animals) and the animals died. The average estimated recovery time of the resistant animals was 245 ± 277 days, and the estimated average survival time of the non-resistant Iberian ibex was 121 ± 71 days. Non-resistant animals survived at least 100 days, while all of them died with less than 200 days. Sixty per cent of the resistant animals were recovered with less than 200 days. Conclusions We report, for the first time, the successful use of radio collars for wildlife diseases monitoring using Iberian ibex/S. scabiei as a model. By using radio collars we documented that most of the Sarcoptes-infected Iberian ibex are resistant to this disease, and we estimate

Capturing wild animals can be time consuming and difficult or even impractical. Noninvasive sampling is potentially a cost-effective and efficient means to monitor wild animals, thereby avoiding the need of capture and disturb species in the wild. On the basis of the morphological and genetic analyses of owl pellet contents, a so far undetected European snow vole (Chionomys nivalis) population was discovered in the Sierra Segura mountain range (Southern Spain). The mtDNA sequence from the newly discovered haplotype was compared with sequences from vole populations of the Sierra Nevada and Sierra Peñalara mountain ranges (Spain) and from Churwalden (Switzerland). The nine recovered haplotypes clustered in four distinct lineages according to their geographical origin. The vole sequence from the Sierra Segura owl pellet belonged to a new haplotype, constituting a new lineage. The evolutionary divergence between sequences from the Sierra Segura and other Spanish populations was higher than that among other Spanish haplotypes. The new snow vole haplotype from this new locality duplicates the number of occurrence sites of this critically endangered species in Southern Spain, which is of great interest for further conservation and management plans of the European snow vole in the most southwestern area of its entire distribution range. © 2013 Informa UK, Ltd.

[Background]: During the last decades, attempts have been made to understand the molecular epidemiology of Sarcoptes scabiei, and to detect and clarify the differences between isolates from different hosts and geographic regions. Two main phenomena have been described: (i) host-taxon derived-Sarcoptes mite infection in European wild animals (revealing the presence of three separate clusters, namely herbivore-, carnivore- and omnivore-derived Sarcoptes populations in Europe) and (ii) prey-to-predator Sarcoptes mite infection in the Masai Mara ecosystem., [Results]: Using one multiplex of 9 microsatellite markers and Sarcoptes mite samples from sympatric Pyrenean chamois, red deer, red fox and Iberian wolf, different population structure analyses revealed concordance with the host-taxon law described for wild animals in Europe, with two main host-derived Sarcoptes mite populations, herbivore- and carnivore-derived. Surprisingly, Iberian wolf derived Sarcoptes populations had the highest genetic diversity among the other populations, including two different subpopulations: one similar to the herbivore-derived Sarcoptes populations, and another similar to carnivore (fox)-derived Sarcoptes mite population., [Conclusions]: The host-taxon effect in wild animals is still supported with the maintenance of carnivore- and herbivore-derived Sarcoptes clusters’ separation in analyzed mites. However, this phenomenon could be modified with the inclusion of a large predator as wolf in the present work, revealing prey-to-predator Sarcoptes mite infection between the studied host-taxa and suggesting the importance of wolf's immune system for explaining the high variability reported in C. lupus derived mites. Further studies of host diet, behavior and movement, and regarding the role played by its immune system, would be of great help to clarify interactions between the two hypotheses, host-taxon and prey-to-predator.

Background: Very few studies have ever focused on the elephants that are wounded or killed as local communities attempt to scare these animals away from their settlements and farms, or on the cases in which local people take revenge after elephants have killed or injured humans. On the other hand, local communities live in close proximity to elephants and hence can play a positive role in elephant conservation by informing the authorities of the presence of injured elephants. Methodology/Principal Findings: Between 2007 and 2011, 129 elephants were monitored in Masai Mara (Kenya), of which 54 had various types of active (intentionally caused) or passive (non-intentionally caused) injuries. Also studied were 75 random control samples of apparently unaffected animals. The observed active injuries were as expected biased by age, with adults suffering more harm; on the other hand, no such bias was observed in the case of passive injuries. Bias was also observed in elephant sex since more males than females were passively and actively injured. Cases of passive and active injuries in elephants were negatively related to the proximity to roads and farms; the distribution of injured elephants was not affected by the presence of either human settlements or water sources. Overall more elephants were actively injured during the dry season than the wet season as expected. Local communities play a positive role by informing KWS authorities of the presence of injured elephants and reported 43% of all cases of injured elephants. Conclusions: Our results suggest that the negative effect of local communities on elephants could be predicted by elephant proximity to farms and roads. In addition, local communities may be able to play a more positive role in elephant conservation given that they are key informants in the early detection of injured elephants.

Dispersal has consequences not only for individual fitness, but also for population dynamics, population genetics and species distribution. Social Hymenoptera show two contrasting colony reproductive strategies, dependent and independent colony foundation modes, and these are often associated to the population structures derived from inter and intra- population gene flow processes conditioned by alternative dispersal strategies. Here we employ microsatellite and mitochondrial markers to investigate the population and social genetic structure and dispersal patterns in the ant Cataglyphis emmae at both, local and regional scales. We find that C. emmae is monogynous and polyandrous. Lack of detection of any population viscosity and population structure with nuclear markers at the local scale suggests efficient dispersal, in agreement with a lack of inbreeding. Contrasting demographic differences before and during the mating seasons suggest that C. emmae workers raise sexuals in peripheric nest chambers to reduce intracolonial conflicts. The high genetic differentiation recovered from the mtDNA haplotypes, together with the significant correlation of such to geographic distance, and presence of new nuclear alleles between areas (valleys) suggest long-term historical isolation between these regions, indicative of limited dispersal at the regional scale. Our findings on the ecological, social and population structure of this species increases our understanding of the patterns and processes involved under independent colony foundation.

Background For centuries, immature stages of Dipterans have infested humans and animals, resulting in a pathological condition referred to as myiasis. Myiases are globally distributed but they remain neglected diseases in spite of the great medical and veterinary importance. Moreover, there is a paucity of information on the clinical-pathology and/or epidemiology of the infestation, especially in African free ranging wildlife. Findings In the present study we report for the first time an outbreak of traumatic cutaneous myiasis (caused by Old World screwworm, Chrysomyia bezziana and blowfly, Lucilia sp.) in free-ranging common elands (Taurotragus oryx). The infestation affected both animal sexes and different age classes, and had a negative impact on individual fitness as well as the overall health. Severely affected individuals were euthanized, while others were clinically treated, and apparently recovered. Conclusions This study indicates that myiasis-causing flies still exist in Kenya and are able to cause severe outbreaks of clinical cutaneous myiasis in wild animals. The status of these parasites in Kenya, which are of zoonotic potential, are either unknown or neglected.

Social media introduce pivotal changes to communication between individuals, organizations and communities. A clear example of the power of social media is the spread of the revolutionary outbreaks in the Arabic countries during 2011, where people used Facebook, YouTube and Skype to communicate, organise meetings and protest actions. Here I report how Doctor-Activists use these social media as an alarm system for ‘war disease’ outbreaks in the Syrian Spring. Social media are used as an alarm system to attract the attention of international organizations, which should assume their responsibilities and play their part in controlling the outbreak of such war diseases.

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.

The Lesser Antilles is a biodiversity hot spot but unfortunately human disturbance has taken its toll, causing dramatic population declines and even extinction of numerous endemic species. Nevertheless, today the rediscovery of previously thought extinct species is not uncommon. Often, old museum specimens and their original descriptions are the only information available for such species. The application of molecular phylogenetic relationships to extant species can help to elucidate pivotal information on their ecology and conservation. Erythrolamprus cursor is possibly an extinct colubrid racer from Martinique, currently classified as critically endangered on the International Union for Conservation of Nature Red List. Mitochondrial deoxyribonucleic acid sequences were obtained from four E. cursor specimens from the Muséum national d'Histoire naturelle (Paris) collections. All sequences recovered the same haplotype and the level of divergence between E. cursor and E. juliae, from the nearby island of Dominica, was lower than between other intraspecific distances within other Erythrolamprus. Furthermore, phylogenetic analyses confirm that these two species are sister taxa and share most recent common ancestry. We discuss that published ecological data available for the sister species (E. juliae) may help to elucidate information on this species' natural history, ultimately having important implications for a future conservation management program if E. cursor is to be found. We emphasize the urgent need to conduct an exhaustive survey on the supposed last population of E. cursor at Diamond Rock to establish the survival of this species there, to understand how it may have adapted to such an ecosystem, especially in sympatry of several introduced rodent species

Parasite presence in any ecosystem generates complex navigating webs (Parasite-NW) within the system, through which parasites move from one to another host. The appropriate assimilation of parasite navigating web is pivotal for a better understanding of pathogen flow in the ecosystem, with implications for disease control. Sarcoptes mite has been approached from medical, veterinary, entomological, physiological and, recently, molecular sides, to understand its epidemiological navigating web between isolates from different hosts and geographical regions. The obtained conclusions are still a matter of debate. Sarcoptes navigating web (Sarcoptes-NW) is intricate and uncertain, with unexplainable pathogenic flow. In this review we summarize by which routes, under what conditions and at what levels the Sarcoptes mite moves among its hosts.

BackgroundOne of the main aims of forensic investigation is the detection and location of people and substances of interest, such as missing people and illegal drugs. Dogs (Canis lupus var. familiaris) have had an important role in legal and forensic investigations for decades; nonetheless canines’ keen sense of smell has never been utilized in either the surveillance or control of wildlife diseases. The rapid removal and treatment of infected carcasses and/or sick animals is a key task in the management of infectious diseases, but it is usually difficult or impractical to carry out in the wild.ResultsIn this paper we report on a study running over a period of 15 years, in which - for the first time to our knowledge - two disease-detector dogs were trained to follow the scent of Sarcoptes-infected animals and to find carcasses, even under the snow, and apparently no false positives were detected in fieldwork. Sarcoptic mange-detector dogs were used to collect the carcasses of 292 mangy wild animals and to identify, separate from their herd, and capture 63 mange-infected wild animals in the Italian Alps.ConclusionsProperly trained disease-detector dogs are an efficient and straightforward tool for surveillance and control of sarcoptic mange in affected wild animal populations.

Background Hybridization between closely related wild and domestic species is of great concern because it can alter the evolutionary integrity of the affected populations. The high allelic variability of Major Histocompatibility Complex (MHC) loci usually excludes them from being used in studies to detect hybridization events. However, if a) the parental species don’t share alleles, and b) one of the parental species possesses an exceptionally low number of alleles (to facilitate analysis), then even MHC loci have the potential to detect hybrids. Results By genotyping the exon2 of the MHC class II DRB1 locus, we were able to detect hybridization between domestic goats (Capra hircus) and free-ranging Iberian ibex (Capra pyrenaica hispanica) by molecular means. Conclusions This is the first documentation of a Capra pyrenaica × Capra hircus hybridization, which presented us the opportunity to test the applicability of MHC loci as new, simple, cost-effective, and time-saving approach to detect hybridization between wild species and their domesticated relatives, thus adding value to MHC genes role in animal conservation and management.

Background: Pastoralists in low-income countries usually live in close proximity to their animals and thus represent an important repository of information about livestock disease. Since wild and domestic animals often mix freely whilst grazing, pastoralists are also able to observe first-hand the diseases that are present in wildlife and as such are key informants in disease outbreaks in sylvatic animals. We report here the findings of the first study of the knowledge and role of Masai pastoralists in mange in wildlife and livestock in Masai Mara, Kenya. Methodology/Principal Findings: In this paper we describe the knowledge of mange accrued by 56 Masai pastoralists in Kenya and how they respond to it in both wildlife and livestock. In total, 52 (93%) pastoralists had a clear idea of the clinical appearance of mange, 13 (23%) understood its aetiology and 37 (66%) knew that mites were the causal agent. Thirty-nine (69%) believed that mange cross-infection between domestic and wild animals occurs, while 48 (85%) had observed mange in domestic animals including sheep (77%), goats (57%), dogs (24%) and cattle (14%). The pastoralists had also observed wild animals infected with mange, above all lions (19%), gazelles (14%), cheetahs (12%) and wildebeests (2%). In 68% of cases Masai pastoralists treat mange infection or apply control measures, most commonly via the topical use of acaricides (29%) and/or the reporting of the outbreak to the veterinary authorities (21%). In the period 2007–2011, Kenya Wildlife Service received 24 warnings of 59 wild animals with mange-like lesions from the Masai Mara pastoralist community. The reported species were cheetah, lion, wild dog, Thomson’s gazelle and wildebeest. Conclusion: Masai pastoralists have good knowledge of mange epidemiology and treatment. Their observations and the treatments they apply are valuable in the control of this disease in both wild and domestic animals

Background Habitat and food supply loss and disruption, together with man’s pursuit of the animal’s unique horn pose significant threats to the charismatic rhinoceros. Filarial worms have been thought to cause cutaneous lesions in black rhinoceros (Diceros bicornis) in Kenya and South Africa, but never in white rhinoceros (Ceratotherium simum) in the wild, despite the fact that the two species live often in close proximity. Stephanofilaria dinniki has been implicated in the past as the causal agents for such lesions. Findings In this paper we report a putative filariosis outbreak in both black and white rhinos at Meru National Park in Kenya. Four black and five white rhinos were affected by various degrees of filarioid-like lesions, while apparently all sympatric wild and domestic animals were filarial worm-free. Affected rhinos were captured and successfully treated. Comparison between the epidemiological aspects of white and black rhinoceros filariosis, and the possible relations between this outbreak and annual seasons, the presence of oxpeckers and other host species are discussed. Conclusions Our study highlights (i) that filarial infection is not restricted to black rhinos, but it affects both rhinoceros species, and (ii) the importance of the earlier detection and immediate treatment (capture-treat and release) of filarioid infections, which is of pivotal interest for wildlife conservation, and especially the endangered and isolated white and black rhinoceros populations.

The ubiquitous Sarcoptes mite is unexplainable emerging and re-emerging parasite, threat- ening 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 result- ing 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 success- fully treated. The possible relations between this outbreak and annual seasons, animal age-classes and sex, and spatial distribution are discussed

This article documents the addition of 238 microsatellite marker loci to the Molecular Ecology Resources Database. Loci were developed for the following species: Alytes dickhilleni, Arapaima gigas, Austropotamobius italicus, Blumeria graminis f. sp. tritici, Cobitis lutheri, Dendroctonus ponderosae, Glossina morsitans morsitans, Haplophilus subterraneus, Kirengeshoma palmata, Lysimachia japonica, Macrolophus pygmaeus, Microtus cabrerae, Mytilus galloprovincialis, Pallisentis (Neosentis) celatus, Pulmonaria officinalis, Salminus franciscanus, Thais chocolata and Zootoca vivipara. These loci were cross-tested on the following species: Acanthina monodon, Alytes cisternasii, Alytes maurus, Alytes muletensis, Alytes obstetricans almogavarii, Alytes obstetricans boscai, Alytes obstetricans obstetricans, Alytes obstetricans pertinax, Cambarellus montezumae, Cambarellus zempoalensis, Chorus giganteus, Cobitis tetralineata, Glossina fuscipes fuscipes, Glossina pallidipes, Lysimachia japonica var. japonica, Lysimachia japonica var. minutissima, Orconectes virilis, Pacifastacus leniusculus, Procambarus clarkii, Salminus brasiliensis and Salminus hilarii.

The present study examined sequence variability in a portion of the mitochondrial cytochrome c oxidase subunit 1 (pcox1) and NADH dehydrogenase subunits 4 and 5 (pnad4 and pnad5) among 39 isolates of Fasciola spp., from different hosts from China, Niger, France, the United States of America, and Spain; and their phylogenetic relationships were re-constructed. Intra-species sequence variations were 0.0–1.1% for pcox1, 0.0–2.7% for pnad4, and 0.0–3.3% for pnad5 for Fasciola hepatica; 0.0–1.8% for pcox1, 0.0–2.5% for pnad4, and 0.0–4.2% for pnad5 for Fasciola gigantica, and 0.0–0.9% for pcox1, 0.0–0.2% for pnad4, and 0.0–1.1% for pnad5 for the intermediate Fasciola form. Whereas, nucleotide differences were 2.1–2.7% for pcox1, 3.1–3.3% for pnad4, and 4.2–4.8% for pnad5 between F. hepatica and F. gigantica; were 1.3–1.5% for pcox1, 2.1–2.9% for pnad4, 3.1–3.4% for pnad5 between F. hepatica and the intermediate form; and were 0.9–1.1% for pcox1, 1.4–1.8% for pnad4, 2.2–2.4% for pnad5 between F. gigantica and the intermediate form. Phylogenetic analysis based on the combined sequences of pcox1, pnad4 and pnad5 revealed distinct groupings of isolates of F. hepatica, F. gigantica, or the intermediate Fasciola form irrespective of their origin, demonstrating the usefulness of the mtDNA sequences for the delineation of Fasciola species, and reinforcing the genetic evidence for the existence of the intermediate Fasciola form

Arvicolid mitochondrial genomes evolve faster than in any other mammalian lineage. The genetic diversity exhibited by these rodents contrasts sharply with their phenotypic homogeneity. Furthermore, faecal droppings from Arvicolid rodents of similar body size are almost undistinguishable on the basis of pellet morphology and content. In this study, we advan- taged from their high genetic diversity vs. phenotypic homogeneity to document the applicability of mtDNA extraction from vole droppings for latter identification of such via a rapid and efficient nested PCR-based technique using the threa- tened Microtus cabrerae as a model species. We sequenced the mitochondrial control region from 75 individuals belonging to 11 species of Arvicolinae from Spain, Portugal, Greece and Italy, and an additional 19 sequences from ten Microtus spe- cies from other countries were downloaded from Genbank. Based on these control region sequences, we successfully designed and applied a nested PCR for M. cabrerae-specific and arvicolid-generic mtDNA markers to differentiate Cabre- ra’s vole faecal samples among other species of the Arvicolinae subfamily. Although this study used Cabrera’s vole as a model species, similar techniques based on mtDNA sequences may find a broader applicability for noninvasive genetic conservation of vole species and their populations.

This article documents the addition of 238 microsatellite marker loci to the Molecular Ecology Resources Database. Loci were developed for the following species: Alytes dickhilleni, Arapaima gigas, Austropotamobius italicus, Blumeria graminis f. sp. tritici, Cobitis lutheri, Dendroctonus ponderosae, Glossina morsitans morsitans, Haplophilus subterraneus, Kirengeshoma palmata, Lysimachia japonica, Macrolophus pygmaeus, Microtus cabrerae, Mytilus galloprovincialis, Pallisentis (Neosentis) celatus, Pulmonaria officinalis, Salminus franciscanus, Thais chocolata and Zootoca vivipara. These loci were cross-tested on the following species: Acanthina monodon, Alytes cisternasii, Alytes maurus, Alytes muletensis, Alytes obstetricans almogav- arii, Alytes obstetricans boscai, Alytes obstetricans obstetricans, Alytes obstetricans pertinax, Cambarellus montezumae, Cam- barellus zempoalensis, Chorus giganteus, Cobitis tetralineata, Glossina fuscipes fuscipes, Glossina pallidipes, Lysimachia japonica var. japonica, Lysimachia japonica var. minutissima, Orconectes virilis, Pacifastacus leniusculus, Procambarus clarkii, Salminus brasiliensis and Salminus hilarii

Because size reduced, or bottlenecked, populations are more prone to adverse events, the detection of genetic bottleneck signatures in wildlife species is highly relevant for conservation. Here we applied 11 microsatellite markers to the endangered Siberian tiger (Panthera tigris altaica) using tissue and blood samples of animals from the Primorsky region of the Russian Far East. Excess heterozygosity and mode shift in allele frequencies tests were positive, while the M-ratio test was negative, indicating the like- lihood of a contemporary rather than a historical population bottleneck. The recent genetic population bottleneck could be attributed to the well-documented demographic collapse of the Siberian tiger pop- ulation in the 1940s, when population size hit bottom with 20–30 surviving animals. The mean effective population size Ne was 14 Siberian tigers (CI95 : 12–25 animals), and the effective population size/census size ratio (Ne /N ratio) was 0.028. This is the first molecular evidence of a recent Siberian tiger population bottleneck, which is of great interest for further conservation and management plans of the highly endan- gered largest felid species, while the worryingly low effective population size challenges the optimism for the recovery of the huge Siberian cat.

In an analysis of a microsatellite locus (OLADRB1) linked to the MHC DRB1 gene of Iberian and Alpine ibex (Capra pyrenaica and Carpa ibex), we detected strong linkage disequilibrium between both loci. The allele length polymorphism at OLADRB1 was unambiguously linked to a particular DRB1 allele. This allowed us to develop a DRB- STR matching method for both ibex species. Validation of the DRB-STR matching method was performed in 160 Iberian ibex from Spain and 98 Alpine ibex from Switzerland and Italy. This simple and relatively inexpensive protocol may find wide applications in a variety of research areas (e.g., mate choice, pathogen-driven selection) and in the biological conservation and management of the Western European ibex populations

Background Recently, there have been attempts to understand the molecular epidemiology of Sarcoptes scabiei, to evaluate the gene flow between isolates of S. scabiei from different hosts and geographic regions. However, to our knowledge, a molecular study has not been carried out to assess the molecular diversity and gene flow of Sarcoptes mite in a predator/prey ecosystem. Results Our study revealed an absence of gene flow between the two herbivore (Thomson's gazelle and wildebeest)- and between the two carnivore (lion and cheetah)-derived Sarcoptes populations from Masai Mara (Kenya), which is in discrepancy with the host-taxon law described for wild animals in Europe. Lion- and wildebeest-derived Sarcoptes mite populations were similar yet different from the Thomson's gazelle-derived Sarcoptes population. This could be attributed to Sarcoptes cross-infestation from wildebeest ("favourite prey") of the lion, but not from Thomson's gazelle. The cheetah-derived Sarcoptes population had different subpopulations: one is cheetah-private, one similar to the wildebeest- and lion-derived Sarcoptes populations, and another similar to the Thomson's gazelle-derived Sarcoptes mite population, where both wildebeest and Thomson's gazelle are "favourite preys" for the cheetah. Conclusions In a predator/prey ecosystem, like Masai Mara in Kenya, it seems that Sarcoptes infestation in wild animals is prey-to-predator-wise, depending on the predator's "favourite prey". More studies on the lion and cheetah diet and behaviour could be of great help to clarify the addressed hypotheses. This study could have further ramification in the epidemiological studies and the monitoring protocols of the neglected Sarcoptes mite in predator/prey ecosystems.

Liver flukes belonging to the genus Fasciola are among the causes of foodborne diseases of parasitic etiology. These parasites cause significant public health problems and substantial economic losses to the livestock industry. Therefore, it is important to definitively characterize the Fasciola species. Current phenotypic techniques fail to reflect the full extent of the diversity of Fasciola spp. In this respect, the use of molecular techniques to identify and differentiate Fasciola spp. offer considerable advantages. The advent of a variety of molecular genetic techniques also provides a powerful method to elucidate many aspects of Fasciola biology, epidemiology, and genetics. However, the discriminatory power of these molecular methods varies, as does the speed and ease of performance and cost. There is a need for the development of new methods to identify the mechanisms underpinning the origin and maintenance of genetic variation within and among Fasciola populations. The increasing application of the current and new methods will yield a much improved understanding of Fasciola epidemiology and evolution as well as more effective means of parasite control. Herein, we provide an overview of the molecular techniques that are being used for the genetic characterization, detection and genotyping of Fasciola spp..

[Background]: Implicitly, parasite molecular studies assume temporal genetic stability. In this study we tested, for the first time to our knowledge, the extent of changes in genetic diversity and structure of Sarcoptes mite populations from Pyrenean chamois (Rupicapra pyrenaica) in Asturias (Spain), using one multiplex of 9 microsatellite markers and Sarcoptes samples from sympatric Pyrenean chamois, red deer (Cervus elaphus), roe deer (Capreolus capreolus) and red fox (Vulpes vulpes). [Results]: The analysis of an 11-years interval period found little change in the genetic diversity (allelic diversity, and observed and expected heterozygosity). The temporal stability in the genetic diversity was confirmed by population structure analysis, which was not significantly variable over time. Population structure analysis revealed temporal stability in the genetic diversity of Sarcoptes mite under the host-taxon law (herbivore derived- and carnivore derived-Sarcoptes mite) among the sympatric wild animals from Asturias. [Conclusions]: The confirmation of parasite temporal genetic stability is of vital interest to allow generalizations to be made, which have further implications regarding the genetic structure, epidemiology and monitoring protocols of the ubiquitous Sarcoptes mite. This could eventually be applied to other parasite species.

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. © 2009 Springer-Verlag.

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. © 2009 Elsevier B.V. All rights reserved.

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 404 bp 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. © 2008 Elsevier B.V. All rights reserved.

The objective of the present study was to examine the extent of genetic diversity among Sarcoptes scabiei individuals belonging to different skin subunits of the body from individual mangy hosts. Ten microsatellite primers were applied on 44 individual S. scabiei mites from three mangy Iberian ibexes from Sierra Nevada Mountain in Spain. Dendrograms of the mites from the individual Iberian ibexes, showing the proportion of shared alleles between pairs of individual mites representing three skin subpopulations (head, back, and abdomen subunits), allowed the clustering of some mite samples up to their skin subunits. This genetic diversity of S. scabiei at skin-scale did not have the same pattern in all considered hosts: for the first Iberian ibex (Cp1), only mites from the head subunit were grouped together; in the second individual (Cp2), the clustering was detected only for mites from the abdomen subunit; and for the third one (Cp3), only mites from the back subunit were clustered together. Our results suggest that the local colonization dynamics of S. scabiei would have influenced the nonrandom distribution of this ectoparasite, after a single infestation. Another presumable explanation to this skin-scale genetic structure could be the repeated infestations. To our knowledge, this is the first documentation of genetic structuring among S. scabiei at individual host skin-scale. Further studies are warranted to highlight determining factors of such trend, but the pattern underlined in the present study should be taken into account in diagnosis and monitoring protocols for studying the population genetic structure and life cycle of this neglected but important ectoparasite. © 2008 Springer-Verlag.

The present study adapted the HotSHOT method, a technique which has been successfully applied on different kinds of tissues, to studies of Sarcoptes. Some modifications of this technique were made which allowed the quick preparation of PCR-quality Sarcoptes genomic DNA (gDNA), namely applying sodium hydroxide as a substrate for three cycles of thermal shock, followed by a short incubation and pH adjustment with a Tris solution (HotSHOT Plus ThermalSHOCK). The performance of this technique was tested by amplifying a ~450-bp rDNA fragment of the second internal transcribed spacer (ITS-2) and by multi-locus genotyping using ten microsatellites on 520 individual Sarcoptes samples. No difference in performance was observed between gDNA samples prepared using the HotSHOT Plus ThermalSHOCK technique and those prepared using a commercial kit utilizing proteinase K digestion. The results demonstrated that the HotSHOT Plus ThermalSHOCK technique is time-saving, economic, and easily automatable for the preparation of PCR-quality mite gDNA, which has implications for studying the molecular biology of mites with human and animal health significance. Although tested in the present study using Sarcoptes mites as a model, this technique may find broad applicability in extraction of gDNA from other parasites with small sizes and hard bodies. © 2008 Springer-Verlag.