Mutations as molecular tools : The metabolic-rate dependent molecular clock and DNA barcoding of allied species

Mutation and recombination are the fundamental processes leading to genetic variation in natural populations. This variation forms the raw material for evolution through natural selection and drift. Therefore, studying mutation rates may reveal information about evolutionary histories as well as phylogenetic interrelationships of organisms. In this thesis two molecular tools, DNA barcoding and the molecular clock were examined. In the first part, the efficiency of mutations to delineate closely related species was tested and the implications for conservation practices were assessed. The second part investigated the proposition that a constant mutation rate exists within invertebrates, in form of a metabolic-rate dependent molecular clock, which can be applied to accurately date speciation events. DNA barcoding aspires to be an efficient technique to not only distinguish between species but also reveal population-level variation solely relying on mutations found on a short stretch of a single gene. In this thesis barcoding was applied to discriminate between Hylochares populations from Russian Karelia and new Hylochares findings from the greater Helsinki region in Finland. Although barcoding failed to delineate the two reproductively isolated groups, their distinct morphological features and differing life-history traits led to their classification as two closely related, although separate species. The lack of genetic differentiation appears to be due to a recent divergence event not yet reflected in the beetles molecular make-up. Thus, the Russian Hylochares was described as a new species. The Finnish species, previously considered as locally extinct, was recognized as endangered. Even if, due to their identical genetic make-up, the populations had been regarded as conspecific, conservation strategies based on prior knowledge from Russia would not have guaranteed the survival of the Finnish beetle. Therefore, new conservation actions based on detailed studies of the biology and life-history of the Finnish Hylochares were conducted to protect this endemic rarity in Finland. The idea behind the strict molecular clock is that mutation rates are constant over evolutionary time and may thus be used to infer species divergence dates. However, one of the most recent theories argues that a strict clock does not tick per unit of time but that it has a constant substitution rate per unit of mass-specific metabolic energy. Therefore, according to this hypothesis, molecular clocks have to be recalibrated taking body size and temperature into account. This thesis tested the temperature effect on mutation rates in equally sized invertebrates. For the first dataset (family Eucnemidae, Coleoptera) the phylogenetic interrelationships and evolutionary history of the genus Arrhipis had to be inferred before the influence of temperature on substitution rates could be studied. Further, a second, larger invertebrate dataset (family Syrphidae, Diptera) was employed. Several methodological approaches, a number of genes and multiple molecular clock models revealed that there was no consistent relationship between temperature and mutation rate for the taxa under study. Thus, the body size effect, observed in vertebrates but controversial for invertebrates, rather than temperature may be the underlying driving force behind the metabolic-rate dependent molecular clock. Therefore, the metabolic-rate dependent molecular clock does not hold for the here studied invertebrate groups. This thesis emphasizes that molecular techniques relying on mutation rates have to be applied with caution. Whereas they may work satisfactorily under certain conditions for specific taxa, they may fail for others. The molecular clock as well as DNA barcoding should incorporate all the information and data available to obtain comprehensive estimations of the existing biodiversity and its evolutionary history.

Abundancia, riqueza y diversidad insectil asociada al cultivo de Marango (Moringa oleifera L.).

El marango (Moringa oleífera L.) Es un árbol de la familia Moringácea que crece en el trópico, es un cultivo de gran importancia en Centroamérica. En Nicaragua durante los últimos años a crecido el interés por este cultivo debido a su potencial nutritivo en la alimentación de ganado bovino, porcino y aves. Sin embargo se ha visto afectado por muchos problemas fitosanitarios, causando éstos una drástica reducción en los rendimientos de este rubro. Con el objetivo de generar información para el manejo de estos insectos, realizamos este estudio para identificar los principales insectos asociados al cultivo de marango, describir la fluctuación poblacional de estos insectos en las diferentes épocas del año, calcular la abundancia y su riqueza y el comportamiento de cada uno de ellos. Este trabajo se realizó en el período comprendido de Mayo a Diciembre del 2013 en Managua, en dos fincas experimentales Las Mercedes y CNIA-INTA. En ambas fincas se colocaron trampas para la captura de insectos, trampas de caída libre y trampas de recipientes de plástico de un galón con agua y melaza. El total de trampas por finca fueron 24, los muestreos se realizaron semanalmente y los insectos colectados en el campo fueron llevados al laboratorio para su respectiva identificación. De acuerdo a lo resultados obtenidos, las principales familias de insectos encontrados asociados al cultivo del marango fueron las familias Formicidae, Lonchaeidae, Elateridae, Tachinidae, Syrphidae, Apidae, Coccinellidae, Scarabaeidae, Vespidae, Histeridae, Tenebrionidae y Curculionidae. Se encontró que la mayor abundancia y riqueza de insectos asociados al cultivo de marango fue encontrada en la finca CNIA INTA, también se observó que la familia Formicidae y Lonchaeidae fueron las más abundante en ambas fincas. La mayor Incidencia y Severidad del daño de termitas se encontró en la finca Las Mercedes

A Guide to the freshwater fauna of Ceylon. Supplement 2

This is the second supplement to "A guide to the freshwater fauna of Ceylon" by A. S. Mendis and C. H. Fernando, Bull. Fish. Res. Stn., Ceylon, No. 12, 160 pp. (1962). In the present supplement additions and corrections are made in the sections on Protozoa, Annelida and Arthropoda. The sections on Platyhelrainthes and Nematoda have been expanded and the Acanthocephala added. A list of species recorded and the hosts of the parasitic forms are included. In the "Guide", the insects with only larval stages in aquatic habitats were mentioned only briefly and no species lists were included. In this supplement this gap is largely filled by added notes and inclusion of species lists of all these groups except the Neuroptera, Lepidoptera, Tabanidae, Syrphidae and Stratiomyidae. The orders Neuroptera and Lepidoptera have relatively few members in freshwater habitats and the families Tabanidae, Syrphidae and Stratiomyidae have forms with larvae in aquatic habitats and also in moist places which are not true freshwater habitats. At this time, it is not possible to separate those forms living in freshwater habitats. Short diagnoses of six additional families are given, namely, the ixidae, Psychodidae, Tabanidae Stratiomyidae, Rhagionidae and Sciomyzidae. Keys are provided for the Odonata and Ephermeroptera larvae down to the family level. An attempt has been made to make the references more comprehensive. Works dealing specifically with the Ceylonese fauna are of course included, but in addition those which are of use in diagnosis of local genera and species have been cited.

Entomofauna associated with the olive tree in southern Portugal

Since 2004 several studies have been carried out in order to identify the main insect species that usually inhabiting the olive ecosystem. The field trials have taken place in two olive groves, one situated in Olhão and the other one in Loulé, both in Algarve and also under Integrated Pest Management (IPM). The sampling techniques used differ according to their purpose (sticky traps, pheromone traps, pitfall traps and samples of aerial parts of the trees such as inflorescences, leaves, fruits and branches). Results showed that the main insect pests of olive tree in southern Portugal were the olive fruit fly Bactrocera oleae Gmelin (Diptera: Tephritidae) and the olive moth Prays oleae Bernard (Lepidoptera: Hyponeumetidae). Other insect pests were also found in our olive groves namely the olive psyllid Euphyllura olivina Costa (Homoptera: Psyllidae), the olive dark beetle Phloeotribus scarabaeoides Bernard (Coleoptera: Curculionidae), the mediterranean black scale Saissetia oleae (Olivier) (Homoptera: Coccidae) and the olive thrip Liothripes oleae Costa (Thysanoptera: Phlaeothripidae). Concerning the auxiliary insects that were found in our olives groves they belong to the following orders and families: Diptera (Syrphidae), Coleoptera (Carabidae, Coccinelidae and Staphylinidae), Hemiptera (Anthocoridae and Miridae), Neuroptera (Chrysopidae) and Hymenoptera (Braconidae, Encyrtidae, Eulophidae, Formicidae and Trichogrammatidae).

An ancient pollinator of a contemporary plant (Cyclamen persicum): When pollination syndromes break down

Pollination of Cyclamen persicum (Primulaceae) was studied in two wild populations in Israel. Buzz-pollination proved to be extremely rare, and performed by a large Anthophora bee only. The most frequent pollinators were various unspecialized species of thrips (Thysanoptera) and hoverflies (Syrphidae). In the Winter-flowering populations the commonest visitor was a small primitive moth, Micropteris elegans (Micropterigidae, Lepidoptera). These moths feed on pollen, copulate and oviposit within the flowers. From the rarity of buzz-pollination it is concluded that the genus Cyclamen co-evolved with large bees capable of buzz-pollination, but lost its original pollinators for unknown historical reasons. The vacant niche was then open to various unspecialized pollen consumers such as thrips, hoverflies and small solitary bees. While these insects are not specific to C. persicum and seem to play a minor role only, the moth strictly relies upon Cyclamen and seems to be the most efficient pollinator.

Insects on decomposing carcasses of small rodents in a secondary forest in Southeastern Brazil

The decomposition of small carcasses in the open is frequently neglected although it may provide information of forensic importance. This paper describes an experimental study of arthropod species associated with carcasses of mouse, Mus musculus (Linnaeus, 1758) and rat, Rattus norvegicus (Berkenhout, 1769) (Rodentia: Muridae). Four carcasses were left inside iron cages in sunlit and shady areas in a secondary forest in Southeastern Brazil twice a season for four seasons (n = 16 carcasses of each rodent). The carcasses were removed when arthropods ceased to visit them. The visiting and colonizing invertebrates were collected daily and identified. Immatures were also collected and reared in a laboratory for identification. We collected 6,514 arthropods (820 adults and 5,694 juvenile forms) belonging to 53 species from the families Sarcophagidae, Calliphoridae, Muscidae, Fanniidae, Syrphidae, Richardiidae, Sepsidae, Micropezidae, Otitidae, Drosophilidae, Phoridae, Dolichopodidae, Anthomyiidae, Asilidae and Lauxaniidae (Diptera), Formicidae, Ichneumonidae, Encyrtidae and Apidae (Hymenoptera), Staphylinidae (Coleoptera) and Gonyleptidae (Opiliones). Lucilia eximia (Wiedemann, 1819) (Diptera: Calliphoridae) and Peckia (Pattonella) intermutans (Walker, 1861) (Diptera: Sarcophagidae) deserve special attention because both adult and immature forms were collected in all seasons and in both areas. Our results indicate that the frequency of occurrence of these arthropods was positively associated with carcass size (mouse or rat); no marked insect succession on the carcasses occurred; and the diversity of Calliphoridae and Sarcophagidae was high, irrespective of season.

The opportunistic feeding and reproduction strategies of the annual fish Cynopoecilus melanotaenia (Cyprinodontiformes: Rivulidae) inhabiting ephemeral habitats on southern Brazil

A maioria dos peixes da família Rivulidae são popularmente conhecidos como anuais por completarem todo seu ciclo biológico em pequenos corpos de água temporários que secam obrigatoriamente em determinados períodos do ano causando a morte dos indivíduos adultos. Possuem características biológicas peculiares como pequeno porte, maturação sexual precoce, reprodução contínua, um elaborado padrão de corte e uma grande capacidade reprodutiva entre os peixes. Os rivulídeos se encontram amplamente distribuídos nas Américas do Norte, Central e Sul. Este trabalho analisou a dieta e a biologia reprodutiva de Cynopoecilus melanotaenia. Foram coletados 263 exemplares e a análise de 233 conteúdos gastrointestinais revelou uma dieta invertívora composta principalmente por microcrustáceos (Cladocera, Amphipoda e Ostracoda) e insetos imaturos (Chaoboridae, Culicidae, Syrphidae, mas principalmente larvas de Chironomidae). Foi registrada a lepidofagia na dieta dos machos. A fecundidade foi estimada pela análise de 59 pares de ovários maduros e variou entre 2 a 157 ovócitos (média, 19 ± 26[DP]). A espécie apresentou desova parcelada, uma estratégia para aumentar a chance de sobrevivência a depleções prolongadas. Este trabalho apresenta a primeira investigação sobre a biologia reprodutiva de C. melanotaenia. Os resultados encontrados sugerem a estratégia reprodutiva oportunista de C. melanotaenia e fornecem novas informações biológicas que podem contribuir para pesquisas de conservação e manutenção da espécie em seu ambiente natural.

Diversidade de pulgões e de seus parasitóides e predadores na cultura da alfafa

O objetivo deste trabalho foi verificar a diversidade de pulgões, seus predadores e parasitóides, e a influência de fatores climáticos nas suas populações. Foram realizadas coletas semanais no período de abril/1995 a março/1996, no campo de alfafa da Universidade Federal de Lavras (UFLA), em Lavras, MG. As espécies de pulgões coletadas foram Therioaphis trifolii (Monel) f. maculata, Acyrthosiphon pisum (Harris), A. kondoi Shinji e Aphis craccivora Kock, presentes na cultura durante todo o período de estudo, com picos populacionais em novembro/1995, julho/1995, dezembro/1995 e abril/1996, respectivamente. Foram amostrados insetos predadores das famílias Coccinellidae, Syrphidae, Anthocoridae, Geocoridae e Chrysopidae, tendo as duas últimas ocorrência esporádica. Espécies da família Coccinellidae ocorreram durante todo o período amostral, apresentando o pico populacional no final de dezembro/1995, com precipitação de 20 mm e temperatura de 22,6ºC. A família Syrphidae alcançou maiores números em abril, à precipitação de 53 mm e temperatura de 21ºC. A família Anthocoridae não se manteve por todo o período amostral, porém um pico populacional ocorreu no final de dezembro nas mesmas condições que aquele apresentado pela família Coccinellidae. Os parasitóides da família Aphididae alcançaram pico em junho/1995, à temperatura de 16ºC.

Differentiation of floral color and odor in two fly pollinated species of Metrodorea (Rutaceae) from Brazil

We investigated if differences in morphological characters in two species of Metrodorea (Rutaceae) from Brazilian semideciduous forests correspond to some pollination divergence. M. nigra and M. stipularis are sympatric species, display a similar floral morphology, are protandrous, self-incompatible, their flower periods overlap, and both are pollinated by flies. M. nigra main pollinators are Pseudoptiloleps nigripoda (Muscidae) and Fannia sp. (Fanniidae); M. stipularis major pollinators are Phaenicia eximia (Calliphoridae), Palpada sp. and Ornidia obesa (Syrphidae). The distinct floral odor (disagreeable in M. nigra and sweet in M. stipularis) and color (brownish violet vs. pale yellow) determine the differences on type and number of floral visitors observed. Several species from semideciduous forests initially considered to be pollinated by diverse insects, present flies as main pollinators, stressing the importance of fly pollination in such habitats.

Análise dos processos de decomposição e sucessão ecológica em carcaças de suínos (Sus scrofa L.) mortos por disparo de arma de fogo e overdose de cocaína e protocolo de procedimento diante de corpo de delito

Pós-graduação em Biologia Geral e Aplicada - IBB

Efeito de diferentes sistemas de manejo de plantas invasoras sobre o controle biológico e incidência de Cinara atlantica (Hemiptera: Aphididae) em Pinus taeda e biologia de Coccinelídeos (Coleoptera)

Pós-graduação em Agronomia (Proteção de Plantas) - FCA

Influence of tree hollow characteristics on the diversity of saproxylic insect guilds in Iberian Mediterranean woodlands

Saproxylic diversity assessment is a major goal for conservation strategies in woodlands and it should consider woodland composition and configuration at site and tree level as key modelling factors. However, in Mediterranean woodlands little is known about the relation with the environmental factors that structure their assemblages, especially those linked to tree hollow microhabitats. We assessed the diversity of Syrphidae (Diptera) and Coleoptera saproxylic guilds that co-occurred in tree hollows located in three different Iberian Mediterranean woodlands in the Cabañeros National Park (Spain). Furthermore, we evaluated how differences in tree hollow microenvironmental variables (understood as the physical and biotic characteristics of a hollow and tree individual) influenced saproxylic guild diversity both within and among woodland sites. We found that woodland sites that provided greater heterogeneity of trees and hollow microhabitats determined higher saproxylic guild diversity. Nevertheless, certain species or even complete guilds can be favoured in woodlands where some hollow microhabitats predominate as a consequence of historical tree management. In general, hollow volume was the main determining factor for saproxylic guild richness and abundance in woodland sites, and large hollow volume was usually related to higher diversity, which highlighted the importance of multi-habitat hollow trees. Moreover, saproxylic guilds also responded to other different microenvironmental variables, which indicated different ecological preferences among guilds. The conservation of saproxylic insects in Iberian Mediterranean areas must be addressed to protect woodland sites that provide high diversity and large numbers of tree hollow microhabitats, and practices to enhance microhabitat heterogeneity should even be encouraged.

Where Is My Food? Brazilian Flower Fly Steals Prey from Carnivorous Sundews in a Newly Discovered Plant-Animal Interaction

A new interaction between insects and carnivorous plants is reported from Brazil. Larvae of the predatory flower fly Toxomerus basalis (Diptera: Syrphidae: Syrphinae) have been found scavenging on the sticky leaves of several carnivorous sundew species (Drosera, Droseraceae) in Minas Gerais and São Paulo states, SE Brazil. This syrphid apparently spends its whole larval stage feeding on prey trapped by Drosera leaves. The nature of this plant-animal relationship is discussed, as well as the Drosera species involved, and locations where T. basalis was observed. 180 years after the discovery of this flower fly species, its biology now has been revealed. This is (1) the first record of kleptoparasitism in the Syrphidae, (2) a new larval feeding mode for this family, and (3) the first report of a dipteran that shows a kleptoparasitic relationship with a carnivorous plant with adhesive flypaper traps. The first descriptions of the third instar larva and puparium of T. basalis based on Scanning Electron Microscope analysis are provided.

Diptera danica. Genera and species of flies hitherto found in Denmark.

1.Stratomyiidae Xylophagidae, Coenomyiidae, Tabanidae, Leptididae, Acroceridae.-2. Asilidae, Bombyliidae, Therevidae, Scenopinidae.-3. Empididae.-4. Dolichopodidae.-5. Lonchopteridae, Syrphidae.-6. Pipunculidae, Phoridae.-7. Platypezidae, tachinidae.

Diptera danica : genera and species of flies hitherto found in Denmark /

"Published at the expenses of the Carlsberg fund."