The insect endosymbiont Sodalis glossinidius utilizes a type III secretion system for cell invasion

Sodalis glossinidius is a maternally transmitted secondary endosymbiont residing intracellularly in tissues of the tsetse flies, Glossina spp. In this study, we have used Tn5 mutagenesis and a negative selection procedure to derive a S. glossinidius mutant that is incapable of invading insect cells in vitro and is aposymbiotic when microinjected into tsetse. This mutant strain harbors Tn5 integrated into a chromosomal gene sharing high sequence identity with a type III secretion system invasion gene (invC) previously identified in Salmonella enterica. With the use of degenerate PCR, we have amplified a further six Sodalis inv/spa genes sharing high sequence identity with type III secretion system genes encoded by Salmonella pathogenicity island 1. Phylogenetic reconstructions based on the inv/spa genes of Sodalis and other members of the family Enterobacteriaceae have consistently identified a well-supported clade containing Sodalis and the enteric pathogens Shigella and Salmonella. These results suggest that Sodalis may have evolved from an ancestor with a parasitic intracellular lifestyle, possibly a latter-day entomopathogen. These observations lend credence to a hypothesis suggesting that vertically transmitted mutualistic endosymbionts evolve from horizontally transmitted parasites through a parasitism–mutualism continuum.

Alberti Guilielmi Rothi, medicinae doctoris, physici provincialis ducatus Bremensis, Societatis Halensis Naturae Curiosorum sodalis, Tentamen florae Germanicae.

Tomus III has imprint: In bibliopolio Gleditschiano.

Prevalence and associations of Trypanosoma spp. and Sodalis glossinidius with intrinsic factors of tsetse flies

Trypanosomiasis has been identified as a neglected tropical disease in both humans and animals in many regions of sub-Saharan Africa. Whilst assessments of the biology of trypanosomes, vectors, vertebrate hosts and the environment have provided useful information about life cycles, transmission, and pathogenesis of the parasites that could be used for treatment and control, less information is available about the effects of interactions among multiple intrinsic factors on trypanosome presence in tsetse flies from different sites. It is known that multiple species of tsetse flies can transmit trypanosomes but differences in their vector competence has normally been studied in relation to individual factors in isolation, such as: intrinsic factors of the flies (e.g. age, sex); habitat characteristics; presence of endosymbionts (e.g. Wigglesworthia glossinidia, Sodalis glossinidius); feeding pattern; host communities that the flies feed on; and which species of trypanosomes are transmitted. The purpose of this study was to take a more integrated approach to investigate trypanosome prevalence in tsetse flies. In chapter 2, techniques were optimised for using the Polymerase Chain Reaction (PCR) to identify species of trypanosomes (Trypanosoma vivax, T. congolense, T. brucei, T. simiae, and T. godfreyi) present in four species of tsetse flies (Glossina austeni, G. brevipalpis, G. longipennis and G. pallidipes) from two regions of eastern Kenya (the Shimba Hills and Nguruman). Based on universal primers targeting the internal transcribed spacer 1 region (ITS-1), T. vivax was the predominant pathogenic species detected in flies, both singly and in combination with other species of trypanosomes. Using Generalised Linear Models (GLMs) and likelihood ratio tests to choose the best-fitting models, presence of T. vivax was significantly associated with an interaction between subpopulation (a combination between collection sites and species of Glossina) and sex of the flies (X2 = 7.52, df = 21, P-value = 0.0061); prevalence in females overall was higher than in males but this was not consistent across subpopulations. Similarly, T. congolense was significantly associated only with subpopulation (X2 = 18.77, df = 1, P-value = 0.0046); prevalence was higher overall in the Shimba Hills than in Nguruman but this pattern varied by species of tsetse fly. When associations were analysed in individual species of tsetse flies, there were no consistent associations between trypanosome prevalence and any single factor (site, sex, age) and different combinations of interactions were found to be significant for each. The results thus demonstrated complex interactions between vectors and trypanosome prevalence related to both the distribution and intrinsic factors of tsetse flies. The potential influence of the presence of S. glossinidius on trypanosome presence in tsetse flies was studied in chapter 3. A high number of Sodalis positive flies was found in the Shimba Hills, while there were only two positive flies from Nguruman. Presence or absence of Sodalis was significantly associated with subpopulation while trypanosome presence showed a significant association with age (X2 = 4.65, df = 14, P-value = 0.0310) and an interaction between subpopulation and sex (X2 = 18.94, df = 10, P-value = 0.0043). However, the specific associations that were significant varied across species of trypanosomes, with T. congolense and T. brucei but not T. vivax showing significant interactions involving Sodalis. Although it has previously been concluded that presence of Sodalis increases susceptibility to trypanosomes, the results presented here suggest a more complicated relationship, which may be biased by differences in the distribution and intrinsic factors of tsetse flies, as well as which trypanosome species are considered. In chapter 4 trypanosome status was studied in relation to blood meal sources, feeding status and feeding patterns of G. pallidipes (which was the predominant fly species collected for this study) as determined by sequencing the mitochondrial cytochrome B gene using DNA extracted from abdomen samples. African buffalo and African elephants were the main sources of blood meals but antelopes, warthogs, humans, giraffes and hyenas were also identified. Feeding on multiple hosts was common in flies sampled from the Shimba Hills but most flies from Nguruman had fed on single host species. Based on Multiple Correspondence Analysis (MCA), host-feeding patterns showed a correlation with site of sample collection and Sodalis status, while trypanosome status was correlated with sex and age of the flies, suggesting that recent host-feeding patterns from blood meal analysis cannot predict trypanosome status. In conclusion, the complexity of interactions found suggests that strategies of tsetse fly control should be specific to particular epidemic areas. Future studies should include laboratory experiments that use local colonies of tsetse flies, local strains of trypanosomes and local S. glossinidius under controlled environmental conditions to tease out the factors that affect vector competence and the relative influence of external environmental factors on the dynamics of these interactions.

Classification of echolocation calls from 14 species of bat by Support Vector Machines and Ensembles of Neural Networks

Calls from 14 species of bat were classified to genus and species using discriminant function analysis (DFA), support vector machines (SVM) and ensembles of neural networks (ENN). Both SVMs and ENNs outperformed DFA for every species while ENNs (mean identification rate – 97%) consistently outperformed SVMs (mean identification rate – 87%). Correct classification rates produced by the ENNs varied from 91% to 100%; calls from six species were correctly identified with 100% accuracy. Calls from the five species of Myotis, a genus whose species are considered difficult to distinguish acoustically, had correct identification rates that varied from 91 – 100%. Five parameters were most important for classifying calls correctly while seven others contributed little to classification performance.

Female greater wax moths reduce sexual display behavior in relation to the potential risk of predation by echolocating bats

Female greater wax moths Galleria mellonella display by wing fanning in response to bursts of ultrasonic calls produced by males. The temporal and spectral characteristics of these calls show some similarities with the echolocation calls of bats that emit frequency-modulated (FM) signals. Female G. mellonella therefore need to distinguish between the attractive signals of male conspecifics, which may lead to mating opportunities, and similar sounds made by predatory bats. We therefore predicted that (1) females would display in response to playbacks of male calls; (2) females would not display in response to playbacks of the calls of echolocating bats (we used the calls of Daubenton's bat Myotis daubentonii as representative of a typical FM echolocating bat); and (3) when presented with male calls and bat calls during the same time block, females would display more when perceived predation risk was lower. We manipulated predation risk in two ways. First, we varied the intensity of bat calls to represent a nearby (high risk) or distant (low risk) bat. Second, we played back calls of bats searching for prey (low risk) and attacking prey (high risk). All predictions were supported, suggesting that female G. mellonella are able to distinguish conspecific male mating calls from bat calls, and that they modify display rate in relation to predation risk. The mechanism (s) by which the moths separate the calls of bat and moth must involve temporal cues. Bat and moth signals differ considerably in duration, and differences in duration could be encoded by the moth's nervous system and used in discrimination.

Insight on how fishing bats discern prey and adjust their mechanic and sensorial features during the attack sequence

Several insectivorous bats have included fish in their diet, yet little is known about the processes underlying this trophic shift. We performed three field experiments with wild fishing bats to address how they manage to discern fish from insects and adapt their hunting technique to capture fish. We show that bats react only to targets protruding above the water and discern fish from insects based on prey disappearance patterns. Stationary fish trigger short and shallow dips and a terminal echolocation pattern with an important component of the narrowband and low frequency calls. When the fish disappears during the attack process, bats regulate their attack increasing the number of broadband and high frequency calls in the last phase of the echolocation as well as by lengthening and deepening their dips. These adjustments may allow bats to obtain more valuable sensorial information and to perform dips adjusted to the level of uncertainty on the location of the submerged prey. The observed ultrafast regulation may be essential for enabling fishing to become cost-effective in bats, and demonstrates the ability of bats to rapidly modify and synchronise their sensorial and motor features as a response to last minute stimulus variations.

东亚水鼠耳蝠形态描述与分类

水鼠耳蝠Myotis daubentonii(Chiroptera,Vespertilionidae),广泛分布于欧洲和亚洲,亚种分化众多,在亚洲已报道有M.d.ussuriensis,M.d.loukashkini,M.d.petax和M.d.laniger等,但其分类地位一直受到国内外学者的关注。中国的水鼠耳蝠长期以来被认为属于水鼠耳蝠M.daubentonii亚种。最近有研究认为中国的水鼠耳蝠与欧洲的水鼠耳蝠M.daubentonii不同,并把"petax"提升为种。在中国境内相继采到17只鼠耳蝠标本,根据外形、头骨、牙齿、阴茎骨、线粒体DNA细胞色素b等特征,鉴定为东亚水鼠耳蝠Myotis petax,对中国水鼠耳蝠的种和亚种分类做一讨论。

Chiropteran types I and II interferon genes inferred from genome sequencing traces by a statistical gene-family assembler.

BACKGROUND: The rate of emergence of human pathogens is steadily increasing; most of these novel agents originate in wildlife. Bats, remarkably, are the natural reservoirs of many of the most pathogenic viruses in humans. There are two bat genome projects currently underway, a circumstance that promises to speed the discovery host factors important in the coevolution of bats with their viruses. These genomes, however, are not yet assembled and one of them will provide only low coverage, making the inference of most genes of immunological interest error-prone. Many more wildlife genome projects are underway and intend to provide only shallow coverage. RESULTS: We have developed a statistical method for the assembly of gene families from partial genomes. The method takes full advantage of the quality scores generated by base-calling software, incorporating them into a complete probabilistic error model, to overcome the limitation inherent in the inference of gene family members from partial sequence information. We validated the method by inferring the human IFNA genes from the genome trace archives, and used it to infer 61 type-I interferon genes, and single type-II interferon genes in the bats Pteropus vampyrus and Myotis lucifugus. We confirmed our inferences by direct cloning and sequencing of IFNA, IFNB, IFND, and IFNK in P. vampyrus, and by demonstrating transcription of some of the inferred genes by known interferon-inducing stimuli. CONCLUSION: The statistical trace assembler described here provides a reliable method for extracting information from the many available and forthcoming partial or shallow genome sequencing projects, thereby facilitating the study of a wider variety of organisms with ecological and biomedical significance to humans than would otherwise be possible.

Summer habitat associations of bats between riparian landscapes and within riparian areas

The present study examines those features which promote bat feeding in agricultural riparian areas and the riparian habitat associations of individual species. Activity of Nathusius' pipistrelle (Pipistrellus nathusii), common pipistrelle (Pipistrellus pipistrellus), soprano pipistrelle (Pipistrellus pygmaeus), Leisler's bat (Nyctalus leisleri), and Myotis species (Myotis sp.) were recorded, and their habitat associations both "between" and "within" riparian areas were analyzed. General feeding activity was associated with reduced agricultural intensity, riparian hedgerow provision, and habitat diversity. Significant habitat associations for P. pipistrellus were observed only within riparian areas. Myotis species and P. pygmaeus were significantly related to indices of landscape structure and riparian hedgerow across spatial scales. Myotis species were also related to lower levels of riffle flow at both scales of analysis. The importance of these variables changed significantly, however, between analysis scales. The multi-scale investigation of species-habitat associations demonstrated the necessity to consider habitat and landscape characteristics across spatial scales to derive appropriate conservation plans.

A nocturnal mammal, the greater mouse-eared bat, calibrates a magnetic compass by the sun

Recent evidence suggests that bats can detect the geomagnetic field, but the way in which this is used by them for navigation to a home roost remains unresolved. The geomagnetic field may be used by animals both to indicate direction and to locate position. In birds, directional information appears to be derived from an interaction of the magnetic field with either the sun or the stars, with some evidence suggesting that sunset/sunrise provides the primary directional reference by which a magnetic compass is calibrated daily. We demonstrate that homing greater mouse-eared bats (Myotis myotis) calibrate a magnetic compass with sunset cues by testing their homing response after exposure to an altered magnetic field at and after sunset. Magnetic manipulation at sunset resulted in a counterclockwise shift in orientation compared with controls, consistent with sunset calibration of the magnetic field, whereas magnetic manipulation after sunset resulted in no change in orientation. Unlike in birds, however, the pattern of polarization was not necessary for the calibration. For animals that occupy ecological niches where the sunset is rarely observed, this is a surprising finding. Yet it may indicate the primacy of the sun as an absolute geographical reference not only for birds but also within other vertebrate taxa.

Kinship, parentage and temporal stability in nursery colonies of Leisler’s bat (Nyctalus leisleri)

Levels of genetic relatedness within bat colonies are often unknown, and consequently the reasons for group formation and social organization are unclear. The Leisler's bat (Nyctalus leisleri), like most temperate bat species, forms nursery colonies in summer. We used microsatellite markers to examine identity and to attempt to estimate relatedness among females within a nursery colony, over 2 consecutive years, to ascertain whether females show kinship and natal philopatry, testing the hypothesis that this is the basis of colony formation. Parentage and relatedness of young born within a colony was assessed to investigate mating patterns via male reproductive skew and whether males achieve mating success within their natal colony. While there was evidence for female philopatry, levels of genetic relatedness within colonies were low. This suggests that kinship is not a major determinant in group formation, as roosts also comprise a large number of distant relatives or non-kin. Roost switching and gene flow are likely to be high. Both sexes reproduced in their first year, whereas males appear to be the more dispersive sex. We argue that the physical environment as well as information sharing provided by communal roosting are likely to be important factors for the formation of these large natal colonies in N. leisleri and possibly other lineages of bats. © 2012 The Author.

Prey field switching based on preferential behaviour can induce Lévy flights

Using the foraging movements of an insectivorous bat, Myotis mystacinus, we describe temporal switching of foraging behaviour in response to resource availability. These observations conform to predictions of optimized search under the Lévy flight paradigm. However, we suggest that this occurs as a result of a preference behaviour and knowledge of resource distribution. Preferential behaviour and knowledge of a familiar area generate distinct movement patterns as resource availability changes on short temporal scales. The behavioural response of predators to changes in prey fields can elicit different functional responses, which are considered to be central in the development of stable predator-prey communities. Recognizing how the foraging movements of an animal relate to environmental conditions also elucidates the evolution of optimized search and the prevalence of discrete strategies in natural systems. Applying techniques that use changes in the frequency distribution of movements facilitates exploration of the processes that underpin behavioural changes. © 2012 The Author(s) Published by the Royal Society. All rights reserved.

Empirical assessment of non-invasive population genetics in bats:Comparison of DNA quality from faecal and tissue samples

Non-invasive population genetics has become a valuable tool in ecology and conservation biology, allowing genetic studies of wild populations without the need to catch, handle or even observe the study subjects directly. We address some of the concerns regarding the limitations of using non-invasive samples by comparing the quality of population genetic information gained through DNA extracted from faecal samples and biopsy samples of two elusive bat species, Myotis mystacinus and Myotis nattereri. We demonstrate that DNA extracted from faeces and tissue samples gives comparable results for frequency based population genetic analyses, despite the occurrence of genotyping errors when using faecal DNA. We conclude that non-invasive genetic sampling for population genetic analysis in bats is viable, and although more labour-intensive and expensive, it is an alternative to tissue sampling, which is particularly pertinent when specimens are rare, endangered or difficult to capture. © 2012 Museum and Institute of Zoology PAS.

Behavioural context of multi-scale species distribution models assessed by radio-tracking

Incorporating ecological processes and animal behaviour into Species Distribution Models (SDMs) is difficult. In species with a central resting or breeding place, there can be conflict between the environmental requirements of the 'central place' and foraging habitat. We apply a multi-scale SDM to examine habitat trade-offs between the central place, roost sites, and foraging habitat in . Myotis nattereri. We validate these derived associations using habitat selection from behavioural observations of radio-tracked bats. A Generalised Linear Model (GLM) of roost occurrence using land cover variables with mixed spatial scales indicated roost occurrence was positively associated with woodland on a fine scale and pasture on a broad scale. Habitat selection of radio-tracked bats mirrored the SDM with bats selecting for woodland in the immediate vicinity of individual roosts but avoiding this habitat in foraging areas, whilst pasture was significantly positively selected for in foraging areas. Using habitat selection derived from radio-tracking enables a multi-scale SDM to be interpreted in a behavioural context. We suggest that the multi-scale SDM of . M. nattereri describes a trade-off between the central place and foraging habitat. Multi-scale methods provide a greater understanding of the ecological processes which determine where species occur and allow integration of behavioural processes into SDMs. The findings have implications when assessing the resource use of a species at a single point in time. Doing so could lead to misinterpretation of habitat requirements as these can change within a short time period depending on specific behaviour, particularly if detectability changes depending on behaviour. © 2011 Gesellschaft für ökologie.

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Contient : 1° Extrait du « Livre du Regime des Princes », de « GILLE DE ROMME », traduction [de JEAN GOLEIN] ; 2° « Livre de Viellesse », de « TULLE », traduction de « LAURENT » [DE PREMIERFAIT] ; 3° « Livre de la vraie Amitié », de « TULLE », traduction de « LAURENT DE PREMIERFAIT » ; 4° « Quadrilogium invectivum » [d'ALAIN CHARTIER] ; 5° « Dialogus familiaris amici et sodalis super depploracionem galice calamitatis, ab ALANO AURIGE editus » ; 6° Le Curial [d'ALAIN CHARTIER]