Mites as biological tags of their hosts.

Movements and spatial distribution of host populations are expected to shape the genetic structure of their parasite populations. Comparing the genetic patterns of both interacting species may improve our understanding of their evolutionary history. Moreover, genetic analyses of parasites with horizontal transmission may serve as indicators of historical events or current demographic processes that are not apparent in the genetic signature of their hosts. Here, we compared mitochondrial variation in populations of the ectoparasitic mite Spinturnix myoti with the genetic pattern of its host, the Maghrebian bat Myotis punicus in North Africa and in the islands of Corsica and Sardinia. Mite mitochondrial differentiation among populations was correlated with both host mitochondrial and nuclear differentiation, suggesting spatial co-differentiation of the lineages of the two interacting species. Therefore our results suggest that parasite dispersal is exclusively mediated by host movements, with open water between landmasses as a main barrier for host and parasite dispersal. Surprisingly the unique presence of a continental European mite lineage in Corsica was inconsistent with host phylogeographical history and strongly suggests the former presence of European mouse-eared bats on this island. Parasites may thus act as biological tags to reveal the presence of their now locally extinct host.

Signs of a vector's adaptive choice: on the evasion of infectious hosts and parasite-induced mortality

Laboratory and field experiments have demonstrated in many cases that malaria vectors do not feed randomly, but show important preferences either for infected or non-infected hosts. These preferences are likely in part shaped by the costs imposed by the parasites on both their vertebrate and dipteran hosts. However, the effect of changes in vector behaviour on actual parasite transmission remains a debated issue. We used the natural associations between a malaria-like parasite Polychromophilus murinus, the bat fly Nycteribia kolenatii and a vertebrate host the Daubenton's bat Myotis daubentonii to test the vector's feeding preference based on the host's infection status using two different approaches: 1) controlled behavioural assays in the laboratory where bat flies could choose between a pair of hosts; 2) natural bat fly abundance data from wild-caught bats, serving as an approximation of realised feeding preference of the bat flies. Hosts with the fewest infectious stages of the parasite were most attractive to the bat flies that did switch in the behavioural assay. In line with the hypothesis of costs imposed by parasites on their vectors, bat flies carrying parasites had higher mortality. However, in wild populations, bat flies were found feeding more based on the bat's body condition, rather than its infection level. Though the absolute frequency of host switches performed by the bat flies during the assays was low, in the context of potential parasite transmission they were extremely high. The decreased survival of infected bat flies suggests that the preference for less infected hosts is an adaptive trait. Nonetheless, other ecological processes ultimately determine the vector's biting rate and thus transmission. Inherent vector preferences therefore play only a marginal role in parasite transmission in the field. The ecological processes rather than preferences per se need to be identified for successful epidemiological predictions.

Detection of Pneumocystis in lungs of bats from Brazil by PCR amplification

Pneumocystis has been isolated from a wide range of unrelated mammalian hosts, including humans, domestic and wild animals. It has been demonstrated that the genome of Pneumocystis of one host differs markedly from that of other hosts. Also, variation in the chromosome and DNA sequence of Pneumocystis within a single host species has been observed. Since information about the occurrence and nature of infections in wild animals is still limited, the objective of this work was to detect the presence of Pneumocystis sp. in lungs of bats from two states from Brazil by Nested-PCR amplification. The bats, captured in caves and in urban areas, were obtained from the Program of Rabies Control of two States in Brazil, Mato Grosso and Rio Grande do Sul, located in the Mid-Western and Southern regions of the country, respectively. DNAs were extracted from 102 lung tissues and screened for Pneumocystis by nested PCR at the mtLSU rRNA gene and small subunit of mitochondrial ribosomal RNA (mtSSU rRNA). Gene amplification was performed using the mtLSU rRNA, the primer set pAZ102H - pAZ102E and pAZ102X - pAZY, and the mtSSU rRNA primer set pAZ102 10FRI - pAZ102 10R-RI and pAZ102 13RI - pAZ102 14RI. The most frequent bats were Tadarida brasiliensis (25), Desmodus rotundus (20), and Nyctinomops laticaudatus (19). Pneumocystis was more prevalent in the species Nyctinomops laticaudatus (26.3% = 5/19), Tadarida brasiliensis (24% = 6/25), and Desmodus rotundus (20% = 4/20). Besides these species, Pneumocystis also was detected in lungs from Molossus molossus (1/11, 9.1%), Artibeus fimbriatus (1/1, 100%), Sturnira lilium (1/3, 33.3%), Myotis levis (2/3, 66.7%)and Diphylla ecaudata (1/2, 50%). PCR products which could indicate the presence of Pneumocystis (21.56%) were identified in DNA samples obtained from 8 out of 16 classified species from both states (5 bats were not identified). This is the first report of detection of Pneumocystis in bats from Brazil.

Food webs in the era of molecular revolution – Like resolving the Gordian knot with a tricorder

Living nature consists of countless organisms, which are classified into millions of species. These species interact in many ways; for example predators when foraging on their prey, insect larvae consuming plants, and pathogenic bacteria drifting into humans. In addition, abiotic nature has a great initiative impact on life through many factors (including sunlight, ambient temperature, and water. In my thesis, I have studied interactions among different life forms in multifaceted ways. The webs of these interactions are commonly referred to as food webs, describing feeding relationships between species or energy transfer from one trophic level to another. These ecological interactions – whether they occur between species, between individuals, or between microorganisms within an individual – are among the greatest forces affecting natural communities. Relationships are tightly related to biological diversity, that is, species richness and abundances. A species is called a node in food web vocabulary, and its interactions to other species are called links. Generally, Artic food webs are considered to be loosely linked, simple structures. This conception roots into early modern food webs, where insects and other arthropods, for example, were clumped under one node. However, it has been shown that arthropods form the greatest part of diversity and biomass both in the tropics and in Arctic areas. Earlier challenges of revealing the role of insects and microorganisms in interactions webs have become possible with the help of recent advances in molecular techniques. In the first chapter, I studied the prey diversity of a common bat, Myotis daubentonii, in southwestern Finland. My results proved M. daubentonii being a versatile predator whose diet mainly consists of aquatic insects, such as chironomid midges. In the second chapter, I expanded the view to changes in seasonal and individual-based variation in the diet of M. daubentonii including the relationship between available and observed prey. I found out that chironomids remain the major prey group even though their abundance decreases in proportion to other insect groups. Diet varied a lot between individuals, although the differences were not statistically significant. The third chapter took the study to a large network in Greenland. I showed that Artic food webs are very complex when arthropods are taken into account. In the fourth chapter, I examined the bacterial flora of M. daubentonii and surveyed the zoonotic potential of these bacteria. I found Bartonella bacteria, of which one was described as a new species named after the locality of discovery. I have shown in my thesis that Myotis daubentonii as a predator links many insect species as well as terrestrial and aquatic environments. Moreover, I have exposed that Arctic food webs are complex structures comprising of many densely linked species. Finally, I demonstrated that the bacterial flora of bats includes several previously unknown species, some of which could possibly turn in to zoonosis. To summarize, molecular methods have untied several knots in biological research. I hope that this kind of increasing knowledge of the surrounding nature makes us further value all the life forms on earth.

Reduction of a native fish fauna by alien species: an example from Brazilian freshwater tropical lakes

The consequences of introducing Cichla cf. monoculus Spix & Agassiz, Astronotus ocellatus (Agassiz) and Pygocentrus nattereri Kner into lakes in the River Doce basin, Brazil, on richness, diversity and efficiency of aquatic macrophytes as natural refugia to native fishes was investigated. Samples were taken from lakes with and without alien fishes in areas with and without aquatic macrophytes. The presence of alien fishes reduced richness and diversity of the native fish community. The refugia function, which could be attributed to the clustering of aquatic macrophytes, does not exist in these lakes probably because the alien fishes exploit such habitats for reproduction. Since introductions threaten the native fish diversity of the region, studies on regional dispersion and factors that minimise the spread of alien fishes are needed.

Spatio-temporal segregation and size distribution of fish assemblages as related to non-native species occurrence in the middle rio Doce Valley, MG, Brazil

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Similarity of ground-dwelling anuran (Amphibia) composition among different vegetation physiognomies in a Mesophytic Semideciduous Forest from southeastern Brazil

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Seasonal variation in body size of tropical anuran amphibians.

Most anurans are considered short lived reproducing only once, with adult size being an important factor contributing to mating success. We sampled anurans with 30 pitfall traps in three forest patches of the northwestern São Paulo State, Brazil, throughout one year. Our objective was to analyze the variation in anuran body size between the dry and rainy season. Our hypothesis is that anurans registered during the rainy season will be larger than conspecifics registered during dry season, since there is a selective advantage in having a larger body size during reproduction activity. The most abundant species registered in the patches were Eupemphix nattereri, Physalaemus cuvieri and Leptodactylus podicipinus. Males and females of E. nattereri and P. cuvieri recorded in the rainy season were larger than of those captured in the dry one. In contrast, males of L. podicipinus were larger in the dry season than in the rainy season. Besides, the operational sex ratio (OSR) in the breeding season was biased towards males in these three species with, respectively, 4.66, 3.2 and 1.87 males per female. Our results suggest that probably body size variation between different seasons is a consequence of individuals' turnover between rainy seasons. Lower individuals captured in the dry season, would be reproductively active adults in the next breeding season.

Uso de fragmentos florestais por anuros (Amphibia) de área aberta na região noroeste do Estado de São Paulo

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Anuran captured in pitfall traps in three agrossystem in Northwestern São Paulo State, Brazil

Embora seja geralmente assumido que a agricultura influencia negativamente populações de anfíbios, existem poucos estudos sobre os efeitos dos cultivos agrícolas em anuros neotropicais. Visando contribuir para diminuir essa lacuna de conhecimento, no presente estudo buscamos verificar quais espécies de anuros estão presentes nos agrossistemas. Para isso, usamos dados de anuros capturados em armadilhas de queda, inicialmente proposto para o levantamento da fauna de opiliões em três agrossistemas (milho, soja e seringal). Nós registramos quatro espécies de anuros nas armadilhas de queda: Leptodactulus fuscus, L. mystacinus (Leptodactylidae), Eupemphix nattereri e Physalaemus cuvieri (Leiuperidae). Na plantação de milho foram registradas quatro espécies e 30 indivíduos, no seringal quatro espécies e 11 indivíduos e na plantação de soja três espécies e oito indivíduos. Nossos resultados mostram que os anuros estão presentes nos agrossistemas, principalmente espécies de anuros generalistas.

Morcegos cavernícolas da região do Distrito Federal, centro-oeste do Brasil (Mammalia, Chiroptera)

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

The diet of bats from Southeastern Brazil: the relation to echolocation and foraging behaviour

In this study the incidence of moths and beetles was examined from feces samples of bats that use different foraging behaviors. Twenty sites around the Fazenda Intervales, a Field Research Station located in São Paulo State, in southeastern Brazil were sampled. Feces were collected from bats caught in mist nets, Turtle Traps or hand nets and, in one case, from beneath a roost. Feces samples were taken from six species of bats: Micronycteris megalotis (Gray, 1842), Mimon bennettii (Gray, 1838), Furipterus horrens (F. Cuvier, 1828), Myotis riparius Handley, 1960, Myotis ruber (E. Geoffroy, 1806) and Histiotus velalus (I. Geoffroy, 1824). To record and describe the frequencies dominating bat echolocation calls, an Anabat II bat detector coupled with an Anabat ZCA interfaces and DOS laptop computers were used. The data show that Furipterus horrens feeds extensively on moths, as predicted from the features of its echolocation calls. Gleaning bats, whose echolocation calls are much less conspicuous to moths take a wide range of insect (and other) prey.

Riqueza de espécies de morcegos (Mammalia, Chiroptera) em dois diferentes habitats, na região centro-sul do Paraná, sul do Brasil

Estudo preliminar sobre a riqueza de espécies e a freqüência de capturas de morcegos em dois habitats, mata primária e reflorestamento de Araucária, na Região Centro Leste do Paraná. Foi empregado o mesmo esforço para as duas áreas, resultando na captura de 11 espécies na área de mata primária, contra oito no reflorestamento de Araucária. A uniformidade na distribuição da abundância -equitabilidade - foi maior na mata primária. No reflorestamento de Araucária, noventa por cento dos indivíduos coletados pertenciam somente a duas espécies. Pelos resultados, conclui-se que a floresta primária com numerosos nichos, apresenta uma riqueza de espécies elevada, mas a ação do homem reduz a riqueza de espécies geral, o que se reflete principalmente na dominância de poucas espécies. Destaca-se aqui, nos dois tipos de ambientes, a presença de Myotis ruber, listada como ameaçada de extinção no Brasil.

Bat rabies in the north-northwestern regions of the state of São Paulo, Brazil: 1997-2002

OBJETIVO: Os relatos sobre a ocorrência de raiva em morcegos no Brasil são esporádicos e isolados. Assim, o objetivo do estudo foi descrever a detecção do vírus da raiva em morcegos do Estado de São Paulo. MÉTODOS: Foram analisados 7.393 morcegos provenientes de 235 municípios do norte e noroeste do Estado de São Paulo, no período de 1997 a 2002 e identificados por meio de características morfológicas e morfométricas. Para a detecção do antígeno viral foi utilizada a técnica de imunofluorescência direta e o isolamento do vírus foi realizado por inoculação em camundongos. RESULTADOS: Das amostras examinadas, 1,3% foram positivas para raiva, com variação de 0,2% em 1997 a 1,6% em 2001. Foram encontrados 98 morcegos com o vírus, 87 deles em área urbana. O vírus da raiva foi detectado pela imunofluorescência direta em 77 do total de amostras positivas, enquanto 92 produziram doença em camundongos inoculados e o período de incubação variou entre 4-23 dias. em 43 municípios foi encontrado pelo menos um morcego positivo. Entre as espécies analisadas o vírus da raiva foi detectado com maior freqüência (33,7%) em Artibeus lituratus. Os vespertilionideos do gênero Eptesicus e Myotis totalizaram 24,5% dos morcegos positivos e as espécies do gênero Molossus (Molossus molossus e Molossus rufus), 14,3%. A distribuição do vírus da raiva foi semelhante entre fêmeas (33; 48,5%) e machos (35; 51,5%). CONCLUSÕES: Morcegos positivos para raiva foram encontrados em situações que colocam em risco tanto a população humana como animais de estimação, exigindo medidas voltadas para o manejo destas espécies e de educação da população.

Genetic variability in species of bats revealed by RAPD analysis

Random amplified polymorphic DNA molecular marker was utilized as a means of analyzing genetic variability in seven bat species: Molossus molossus, M. rufus, Eumops glaucinus, E. perotis, Myotis nigricans, Eptesicus furinalis, and Artibeus planirostris. The determination of genetic diversity was based on 741 bands produced by a 20-random primer set. Only eight bands were considered monomorphic to one species. The greatest number of bands and the most polymorphic condition were exhibited by M. molossus, followed by M. nigricans, A. planirostris, E. furinalis, E. glaucinus, M. rufus, and E. perotis. Nei's genetic diversity index in the seven species considering the 20 primers was not greater than 0.22, but some primers were capable of detecting values between 0.39 and 0.49. Nei's unbiased genetic distance values and the UPGMA clustering pattern show that M. molossus and M. rufus have a close genetic relationship, unlike that observed between E. perotis and E. glaucinus. The latter was clustered with A. planirostris and E. furinalis. The low values for genetic diversity and distance observed indicate a genetic conservatism in the seven species. The fluorescent in situ hybridization experiments did not confirm a monomorphic condition for the eight bands identified, demonstrating that the monomorphic bands obtained by random amplified polymorphic DNA are insufficient for the identification of bat species.