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The sanitary control of threatened wild animals is of pivotal interest for their conservation. This task, however, is highly complex in wildlife/livestock systems. In this paper we report findings from a 2-year cross-sectional study of the epidemiology and attempted control of a Sarcoptes mite infestation in the threatened cheetah population in Masai Mara (Kenya), and discuss its interaction with sympatric wild (lion, wildebeest and Thomson's gazelle) and domestic (dog, cattle and sheep) animals. Sarcoptes scabiei was isolated from cheetahs, Thomson's gazelles, wildebeests, lions, cattle, goats and dogs; Psoroptes ovis, on the other hand, was only isolated from sheep. The prevalence study revealed 12·77% infection rates in cheetahs, 4·7% in dogs, 0·8% in Thomson's gazelles, 0·8% in sheep, 0·09% in cattle, and 0·09% in goats, while it opportunistically affected lions and wildebeest. Our study revealed that prevalence of Sarcoptes mite in cheetah population was not associated with the studied geographical blocks, animal sex or the presence of affected domestic animals. Cheetah infection with S. scabiei was associated with the climatic conditions (dry more than wet season) and the balancing between the total number of Thomson's gazelles and the prevalence of infected individuals. Apparently the high prevalence of mangy gazelles has a negative effect on cheetah; this negative effect was reduced when the number of healthy gazelles was increased. Treatment with injectable ivermectin of the clinically affected wild and domestic animals during the first year of this study was associated with much lower incidence of sarcoptic mange during the second year. © Copyright © Cambridge University Press 2012.

[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., This is a contribution to the agreement between CSIC and Principado de Asturias. The research was partially supported by RNM-6400, Proyecto de Excelencia (Junta de Andalucia, Spain), Project RTA 2011-00087-00-00 (INIA), and Juan de la Cierva Grant to Dr S. Alasaad.

Here we report a mammal sexing procedure based on the detection of quantitative differences between females and males in the X-linked loci (quantitative sexing, Q-sexing). This novel technique was validated using samples from Siberian tigers ( Panthera tigris altaica ) whose sexes were known. The Q-sexing technique relies on the fact that amplifications proceeding exclusively from the two X chromosomes in a female mammal should reach the threshold cycle (CT) in a real-time quantitative real time polymerase chain reaction (qPCR) assay sooner than amplifications from the single male X chromosome. Nevertheless, given that the amplification efficiency may vary between samples, results have to be calibrated to a marker that does not vary in copy number between the sexes (for example, an autosomal-linked locus). For this purpose we used quantitative real time polymerase chain reaction (RT qPCR) assays to quantify the amount of three specific Siberian tiger microsatellite markers (X-/Y- and autosomal-linked loci) in individual samples in order to determine the sex of an animal. A difference of one CT between the X and the autosome-linked loci was detected in males, but no such difference was present in female samples. The Q-sexing technique unambiguously separates female from male Siberian tigers. The future of RT qPCR is bright as technology is becoming ever more rapid, cost-effective, easier to use and capable of processing higher throughputs. Thus, we expect that our novel technique for animal sexing will have a wide applicability, although further studies are still needed to adapt it to other animal species using specific primers. Key words : Polymerase chain reaction (PCR), quantitative real time polymerase chain reaction (qPCR), quantitative sexing, Siberian tiger.

In this study, we analysed the Sarcoptes scabiei infrapopulations of 315 Iberian ibex (Capra pyrenaica) by means of skin digestion and differential mite counts. Mean density reached 178.2 mites/cm (range: 0.71-4514.29). Although the most frequently affected zones were hosts extremities and withers, higher mite densities were obtained from lips and extremities, suggesting that indirect infection through the substrate could also play a role in the disease epidemiology, together with direct transmission. The sex ratio of mites was female-biased (mean: 5.18 ♀♀/♂, range: 0.5-17). Larvae were more frequently found (nearly 50% of specimens) than nymphs, adult females or adult males. Mite density varied significantly between years and decreased with host age. Most of the variance was explained by the year, host sex, weight and kidney fat index (KFI). Taking into account the catabolic nature of the disease, our results suggest that hosts in good body condition could evidence a certain degree of resistance to the disease, which presumably has a genetic basis.

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

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

The objective of the present study was to examine the phylogenetic position of Setaria cervi based on sequences of mitochondrial cytochrome c oxidase subunit 1 (cox1) gene. A fragment of the cox1 gene from two morphologically identified S. cervi collected from red deer (Cervus elaphus) from Italy were amplified, sequenced, and compared with corresponding sequences of other filarioid nematode species. Phylogenetic studies using Bayesian analysis revealed S. cervi as monophyletic with other Setaria species, confirming S. cervi as a member of the Setaria genus. S. cervi appeared to be sister species to Setaria labiatopapillosa and Setaria digitata. Setaria tundra and Setaria equina, the other two Setaria species presented in the Italian fauna, formed a sister group to the clade consisting of S. cervi, S. labiatopapillosa, and S. digitata. In addition to phylogenetic clarification, our study is the first molecular identification of S. cervi, which may be useful for further molecular identification and differen- tiation of this filarial worm from other filarioid nematode species, especially in the earlier developmental stages of its life cycle

Heterogeneity in host compatibility is one of the main hypotheses proposed to explain uneven resistance to parasites and uneven parasite load between hosts. It suggests that differences between hosts modulate their predispositions as suitable environments for their potential parasites. Interesting studies of antiparasitic behavior have reported the existence of behavioral traits that are capable of removing foreign particles and of reducing the success of parasitic infections. These traits favor host neatness, although little is known about the heterogeneity of neatness. We used a standardized pseudoinfection with pseudoectoparasites (PEPs) to test the effects of sex, age, and season on the loss of PEPs by hosts as a means of exploring the factors determining neatness in the Iberian ibex Capra pyrenaica. Behavioral observations were also performed to analyze investment in antiparasitic behavior in terms of sex, age, and season. The life span of PEPs peaked in the period December--January, decreased with host age, and was longer in females than in males. Investment in antiparasitic behavior is also associated with both sex and age and season but in a different pattern with interactions between such factors. Our results disagree with the hypothesis that small-bodied animals should be less compatible to carry contact-transmitted particles, such as ectoparasites, in comparison with larger animals. This preexisting hypothesis is thus an inadequate way of predicting host neatness. Consequently, our experiment underlines the importance that nonimmunological traits play in determining heterogeneity in host compatibility to contact-transmitted foreign bodies and helps improve understanding of neatness and of host--parasite systems. Copyright 2011, Oxford University Press.

The present study aimed to establish a fluorescence- based polymerase chain reaction-linked single-strand conformation polymorphism (F-PCR-SSCP) assay for the identification of Fasciola spp. Based on the sequences of the second internal transcribed spacer (ITS-2) of the nuclear ribosomal DNA, we designed a set of genus- specific primers for the amplification of Fasciola ITS-2, with an estimated size of 140 bp. These primers were labelled by fluorescence dyes, and the PCR products were analyzed by capillary electrophoresis under non- denaturing conditions (F-PCR-SSCP). Capillary electropho- resis analysis of the fluorescence-labelled DNA fragments displayed three different peak profiles that allowed the accurate identification of Fasciola species: one single peak specific for either Fasciola hepatica or Fasciola gigantica and a doublet peak corresponding to the “intermediate” Fasciola. Validation of our novel method was performed using Fasciola specimens from different host animals from China, Spain, Nigeria, and Egypt. This F-PCR-SSCP assay provides a rapid, simple, and robust tool for the identification and differentiation between Fasciola spp.

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 advantaged 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 threatened 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 species 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 Cabrera’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.

a b s t r a c t Ten markers specific to Sarcoptes mites were used in applying microsatellite genotyping to individual Sar- coptes mites collected in three European countries from 15 wild mammal populations belonging to 10 host species. The results showed that geographical separation had real biological significance for the def- inition of mite sub-populations, and that the degree of genetic exchange occurring between mites from different localities was apparently related to the geographical distance between locations. Wild host- derived mite populations were found to be clustered into three main groups: herbivore-, carnivore- and omnivore-derived Sarcoptes populations, with the omnivore-derived group located halfway between the herbivore- and carnivore-derived Sarcoptes populations. The separation between these three mite groups was better supported than the geographical separations; nevertheless, a kind of sub-clustering was detected within each of these three groups that separates mite populations into their geographical localities (countries). The lack of gene flow between Sarcoptes populations may have improved parasitic adaptations and led to what we refer to as a host-taxon-derived (carnivore host-, herbivore host- and omnivore host-derived) Sarcoptes mite found on European wild animals. Our results demonstrate that Sarcoptes is not a single panmictic population, even within each geographical location. This finding will have important ramifications for the study of the genetic structure of populations, life cycles, diagnosis and the monitoring protocols of the ubiquitous Sarcoptes mite, and could thus contribute to a better understanding of its associated epidemiology, which is of pivotal interest for wildlife biological conservation.

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.

A collection of483 samples representing Fasciola from six naturally infected host species and 16 localities in Spain, previously identified morphologically and genetically as Fasciola hepatica, was characterized by a novel genetic marker, namely sequence-related amplified polymorphism (SRAP), aiming to reveal genetic variability within F. hepatica in Spain. Visualization of amplification fragments was carried out on 6% denaturing polyacrylamide gels, followed by staining with 0.1% AgNO3 solution. Ten SRAP primer combinations were tested--six of them turned out to be polymorphic. Thirty-four representative F. hepatica samples from six host species and 16 geographical localities showed polymorphic banding patterns using SRAP primer combinations and were grouped into four major clusters using the unweighted pair-group method with arithmetic averages, indicating the existence of genetic variability within the examined F. hepatica samples. These four clusters were not related to particular host species and/or geographical origins of the samples. The results of the present study revealed that SRAP markers were useful in revealing sufficient polymorphism in F. hepatica samples from Spain and had implications for studying the population genetic structure of the Spanish F. hepatica. To our knowledge, this is the first application of SRAP marker to study genetic variation in parasites of human and animal health significance.

Between 1995 and 2006, we surveyed the presence of Fasciola hepatica in Iberian ibex (Capra pyrenaica) from Andalucía (southern Spain) by both necropsy (n = 2,096) and coprological approaches (n = 380). Most of the samples came from the Sierra Nevada mountain range (n = 1,884 and 267, respectively), and all positive cases involved animals from this location. The prevalence reached 0.53% by necropsy and 1.87% by faecal examination. Taking into account both diagnostic methodologies and the total number of animals affected (n = 14), we obtained a yearly prevalence of 0.7 +/- 0.3%. The infection with F. hepatica was found not to be related to host sex, climatology or to co-infection with Sarcoptes scabiei (the most important parasite affecting Iberian ibex, with a prevalence of 49.27 +/- 7.90% in the examined animals). The prevalence of fasciolosis decreased significantly during the period under study and this would be explained by an increase of ibex resistance to this fluke as a result of a reduction of the parasite abundance in the area and/or a reduction of the host infection rate. There was no statistical difference between the two diagnostic methods for the examination of fasciolosis during the period in which both methods were used. Therefore, examination of faecal samples as a non-invasive procedure may provide a useful approach for monitoring fasciolosis in wild ungulate populations. The results of the present study provided foundation for the effective control of F. hepatica infection in Iberian ibex.

In the present study, 25 samples representing Fasciola (Platyhelminthes: Trematoda: Digenea) from nine host species and 19 geographical locations in Spain were characterized genetically by sequences of the first (ITS-1) and second (ITS-2) internal transcribed spacers (ITS) of nuclear ribosomal DNA (rDNA). The ITS rDNA was amplified from individual liver flukes by polymerase chain reaction (PCR), and the amplicons were sequenced directly. The lengths of the ITS-1 and ITS-2 sequences were 422 and 362 bp, respectively, for all Spanish liver fluke samples sequenced. Comparison of the ITS sequences of the Spanish Fasciola samples examined in the present study with that of Fasciola hepatica, Fasciola gigantica and the "intermediate Fasciola" revealed that all Spanish Fasciola samples examined represent the single species of F. hepatica, with only slight sequence variation in the ITS-2 (1/362, 0.3%) among the sequenced samples, but the sequence variation was not related to particular host species and/or geographical origins of the samples. The Spanish F. hepatica examined differed from Fasciola from elsewhere by two nucleotides in the ITS-2, which provided genetic marker for the differentiation of Spanish F. hepatica from Fasciola from other geographical localities. These results have implications for studying the population genetic structure of the Spanish F. hepatica and for the diagnosis and control of the disease it causes.

[Background] A huge effort in rhinoceros conservation has focused on poaching and habitat loss as factors leading to the dramatic declines in the endangered eastern black rhinoceros (Diceros bicornis michaeli) and the southern white rhinoceros (Ceratotherium simum simum). Nevertheless, the role disease and parasite infections play in the mortality of protected populations has largely received limited attention. Infections with piroplasmosis caused by Babesia bicornis and Theileria bicornis has been shown to be fatal especially in small and isolated populations in Tanzania and South Africa. However, the occurrence and epidemiology of these parasites in Kenyan rhinoceros is not known., [Results] Utilizing 18S rRNA gene as genetic marker to detect rhinoceros infection with Babesia and Theileria, we examined blood samples collected from seven rhinoceros populations consisting of 114 individuals of black and white rhinoceros. The goal was to determine the prevalence in Kenyan populations, and to assess the association of Babesia and Theileria infection with host species, age, sex, location, season and population mix (only black rhinoceros comparing to black and white rhinoceros populations). We did not detect any infection with Babesia in the sequenced samples, while the prevalence of T. bicornis in the Kenyan rhinoceros population was 49.12% (56/114). White rhinoceros had significantly higher prevalence of infection (66%) compared to black rhinoceros (43%). The infection of rhinoceros with Theileria was not associated with animal age, sex or location. The risk of infection with Theileria was not higher in mixed species populations compared to populations of pure black rhinoceros., [Conclusion] In the rhinoceros studied, we did not detect the presence of Babesia bicornis, while Theileria bicornis was found to have a 49.12% prevalence with white rhinoceros showing a higher prevalence (66%) comparing with black rhinoceros (43%). Other factors such as age, sex, location, and population mix were not found to play a significant role., We acknowledge support by the CSIC Open Access Publication Initiative through its Unit of Information Resources for Research (URICI)

[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., This work was supported in part by the International Science & Technology Cooperation Program of China (Grant No. 2013DFA31840), the Science Fund for Creative Research Groups of Gansu Province (Grant No. 1210RJIA006) to XQZ, and the China Postdoctoral Science Foundation (Grant No. 201104363) to MJX.

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

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.

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.

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.

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.

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.

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., Thanks to anonymous Doctor-Activists and Media-Activists, who provided the video and picture materials. Inkad, Al-Susa and Al-Baguz Charity Organizations are thanked for providing drugs for leishmaniasis treatment. Special thanks to the Local Counsels in the studied towns for providing data on leishmaniasis epidemiology and treatment.

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 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., Lukas Keller (Zürich University, Switzerland) is thanked for the helpful comments on the manuscript. Ana Píriz (EBD-CSIC, Seville, Spain) is thanked for help with troubleshooting in the laboratory. The research was supported by Proyecto de Excelencia RNM 06400 (Junta de Andalucia, Spain) and Juan de la Cierva Grant (Ministerio Innovación y Ciencia, Spain).

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

Common names (CN) add to linguistic richness and ultimately derive from how a majority of people refer to a species. CN have a biological and-above all-practical importance given that they are essential for connecting specialists and lay people. To illustrate the diversity of CN between and within species, we made an overview of common name in Caprinae species-flagship species in mountain ecosystems. Then, using Capra pyrenaica as a study case, we highlighted that the choice of CN is inextricably linked to current debates and trends in wildlife management that should never be ignored, given their importance in the fields of ethics, zoology, systematics, conservation and current management. We underline the need to investigate further the probable relationships linking common names, human perception and wildlife management. Researchers, citizens and policy-makers will have to be watchful that clumsy common names, such as 'wild-to-domestic' transformed ones, will not hamper the conservation of wild species as a 'Common Heritage'. © 2011 Springer Science+Business Media B.V.

A cost-effective, reliable and efficient method of obtaining DNA samples is essential in large-scale genetic analyses. This study examines the possibility of using a threatened vole species, Microtus cabrerae, as a model for the collection and preservation of faecal samples for subsequent DNA extraction with a protocol based on the HotSHOT technique. Through the examination of the probability of multi-copies (mitochondrial) and single copy (microsatellite) loci amplification (including the genotype error) and of the DNA yield (estimated by real-time qPCR), the new protocol was compared with both the frequently employed methods that successfully use ethanol to preserve faecal samples and with commercial kit-based DNA extraction. The single-tube HotSHOT-based protocol is a user-friendly, non-polluting, time-saving and inexpensive method of faeces sample collection, preservation and PCR- quality gDNA preparation. This technique therefore provides researchers with a new approach that can be employed in high-throughput, noninvasive genetic analyses of wild animal populations

The development of non-manipulative molecular tools to determine the origin of parasite infections in the animal trade (if infected before their export or import) is of great interest worldwide for both the animal trade industry and for animal welfare. Molecular tools have a wide range of applications, including forensic identification, wildlife preservation and conservation, veterinary public health protection, and food safety. Nonetheless, genetic markers were not reported to detect the source of infection in the animal trade. In this study we tested the applicability of molecular tools to detect the origin of Sarcoptes mite infection of wildebeest imported by the United Arab Emirate (UAE) from Tanzania. Using one multiplex of seven microsatellite markers and control samples from UAE, Kenya and Italy, we demonstrated the usefulness of the multiplex STR-typing as a molecular tool of pivotal interest to help commercialist, authorities, and conservationists, to identify the geographical origin of parasitic infections. © 2011 Springer Science+Business Media, LLC.

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.

[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., The research was supported by a RTA 2009-00114-00-00 (INIA) project, RNM-6400, Proyecto de Excelencia (Junta de Andalucia, Spain) and Juan de la Cierva grant. This work is a contribution to the agreement between IREC - CSIC and Principado de Asturias.

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.., This work was supported, in part, by the National S & T Major Program (Grant No. 2008ZX10004-011, 2009ZX10004-302, 2009ZX10004-201), the National Key Technology R & D Program (Grant No. 2008BAI56B03), the State Key Laboratory of Veterinary Etiological Biology, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, the Yunnan Provincial Program for Introducing High-level Scientists (Grant No. 2009CI125) and the Program for Changjiang Scholars and Innovative Research Team in University (Grant No. IRT0723).

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 advantaged 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 threatened 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 species 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 Cabrera'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.

Noninvasive sampling is a potentially cost-effective and efficient means of monitoring wild animals that precludes the need for captures and avoids undue disturbance. Nevertheless, it is generally difficult to separate faeces deposited by Microtus cabrerae from that of other sympatric rodents of similar body size on the basis of just morphological features and content. Species identification methods in many noninvasive studies involve mtDNA analysis. Here we report the first use of species-specific primers targeting the SRY gene in a noninvasive molecular identification of Cabrera’s vole. This noninvasive molecular-based technique may thus provide us with a potential tool for further genetic and ecological study of this threatened species of Iberian vole.

Real time quantitative PCR (qPCR) is one of the key technologies of the post-genome era, with clear advantages compared to normal end-point PCR. In this paper, we report the first qPCR-based assay for the identification of Fasciola spp. Based on sequences of the second internal transcribed spacers (ITS-2) of the ribosomal rRNA gene, we used a set of genus- specific primers for Fasciola ITS-2 amplification, and we designed species-specific internal TaqMan probes to identify F. hepatica and F. gigantica, as well as the hybrid ‘intermediate’ Fasciola. These primers and probes were used for the highly specific, sensitive, and simple identification of Fasciola species collected from different animal host from China, Spain, Niger and Egypt. The novel qPCR-based technique for the identification of Fasciola spp. may provide a useful tool for the epidemiological investigation of Fasciola infection, including their intermediate snail hosts

Availability of mites is a recognized limiting factor of biological and genetic investigations of the genus Sarcoptes. Current methods of deoxyribonucleic acid (DNA) extraction from individual mites also need substantial improvement in efficiency and operator friendliness. We have first developed a technique for efficient and safe extraction of living mites from scabietic skin samples (crusts or deep skin scrapings). Its core device is a large plastic syringe connected with a 1.5-ml Eppendorf tube. The source material is introduced in the syringe and the device in a shoe box with the tip half of the tube emerging. Mites migrate towards a heat source during a minimum of 36 h. Then, the tube is detached and the mites utilized without risks for the operators. A second technique allows operator-friendly manipulation of individual mites for DNA extraction. Fixed mites are isolated by adhesion to a small strip of polyvinyl chloride (PVC) adhesive tape operated with tweezers. Then, mite and strip are plunged in the lyses buffer and the sample twice submitted to thermal shock for disruption of the chitinous exoskeleton. Data show that the tape does not interfere with successive DNA extraction with a commercial kit. The corresponding protocol, that we briefly name "PVC adhesive tape + thermal shock + kit DNA extraction," compares favorably with the available ones.

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 aim of the present study was to assess whether individual Sarcoptes mites collected from frozen skin ('postponed isolation' method) are suitable sources of PCR-quality genomic DNA, and to test the effectiveness of this method in comparison with the 'direct isolation' method, often used through force of habit. Hundreds of single Sarcoptes scabiei samples, resulting from direct (live) or postponed (post-frozen) isolation, were tested using a ∼450 bp product (ITS-2) and multi-locus 10x genotyping with microsatellite markers. No statistical difference in yield of soluble DNA was found between the two isolation methods. Nevertheless, 19% of the reactions were classified as failed preparations in the direct isolation method, whereas the rate of unsuccessful reactions was 34% in the postponed isolation method. Consequently, post-frozen isolation is suitable and recommendable for Sarcoptes mite gDNA preparation, particularly when performing a balancing act among safety, practicability and profitability. These results have implications for mite collection for DNA extraction, and hence the needed wider leap of Sarcoptes into the genetic era. © 2008 Springer Science+Business Media B.V.

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 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.

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., Project support was provided by Proyecto de Excelencia RNM 06400 (Junta de Andalucia, Spain) and Juan de la Cierva Grant (MinisterioInnovación y ciencia, Spain), the Special Fund for Agro-scientific Research in the Public Interest (Grant No. 201303037), the PRIN 2010/2011 Grant (Ministero Università e Ricerca, Italy, Code D11J2000360001) and the Science Fund for Creative Research Groups of Gansu Province (Grant No. 1210RJIA006).

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