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Phylogenetic niche conservatism (PNC) shapes the distribution of organisms by constraining lineages to parti-cular climatic conditions. Conversely, if areas with similar climates are geographically isolated, diversificationmay also be limited by dispersal. Neotropical xeric habitats provide an ideal system to test the relative roles ofclimate and geography on diversification, as they occur in disjunct areas with similar biotas. Sicariinae sandspiders are intimately associated with these xeric environments, particularly seasonally dry tropical forests(SDTFs) and subtropical deserts/scrublands in Africa (Hexophthalma) and the Neotropics (Sicarius). We explorethe role of PNC, geography and biome shifts in their evolution and timing of diversification. We estimated atime-calibrated, total-evidence phylogeny of Sicariinae, and used published distribution records to estimateclimatic niche and biome occupancy. Topologies were used for estimating ancestral niches and biome shifts. Weused variation partitioning methods to test the relative importance of climate and spatially autocorrelatedfactors in explaining the spatial variation in phylogenetic structure ofSicariusacross the Neotropics. NeotropicalSicariusare ancient and split from their African sister-group around 90 (57–131) million years ago. Most spe-ciation events took place in the Miocene. Sicariinae records can be separated in two groups corresponding totemperate/dry and tropical/seasonally dry climates. The ancestral climatic niche of Sicariinae are temperate/dryareas, with 2–3 shifts to tropical/seasonally dry areas inSicarius. Similarly, ancestral biomes occupied by thegroup are temperate and dry (deserts, Mediterranean scrub, temperate grasslands), with 2–3 shifts to tropical,seasonally dry forests and grasslands. Most of the variation in phylogenetic structure is explained by long-distance dispersal limitation that is independent of the measured climatic conditions. Sicariinae have an ancientassociation to arid lands, suggesting that PNC prevented them from colonizing mesic habitats. However, nichesare labile at a smaller scale, with several shifts from deserts to SDTFs. This suggests that PNC and long-distancedispersal limitation played major roles in confining lineages to isolated areas of SDTF/desert over evolutionaryhistory, although shifts between xeric biomes occurred whenever geographical opportunities were presented

RESUMEN

Abstract Successful animal rearing under laboratory conditions for commercial processes or laboratory experiments is a complex chain that includes several stressors (e.g., sampling and transport) and incurs, as a consequence, the reduction of natural animal conditions, economic losses and inconsistent and unreliable biological results. Since the invasion of the bivalve Limnoperna fortunei (Dunker, 1857) in South America, several studies have been performed to help control and manage this fouling pest in industrial plants that use raw water. Relatively little attention has been given to the laboratory rearing procedure of L. fortunei, its condition when exposed to a stressor or its acclimation into laboratory conditions. Considering this issue, the aims of this study are to (i) investigate L. fortunei physiological responses when submitted to the depuration process and subsequent air transport (without water/dry condition) at two temperatures, based on glycogen concentrations, and (ii) monitor the glycogen concentrations in different groups when maintained for 28 days under laboratory conditions. Based on the obtained results, depuration did not affect either of the groups when they were submitted to approximately eight hours of transport. The variation in glycogen concentration among the specimens that were obtained from the field under depurated and non-depurated conditions was significant only in the first week of laboratory growth for the non-depurated group and in the second week for the depurated group. In addition, the tested temperature did not affect either of the groups that were submitted to transport. The glycogen concentrations were similar to those of the specimens that were obtained from the field in third week, which suggests that the specimens acclimated to laboratory conditions during this period of time. Thus, the results indicate that the air transport and acclimation time can be successfully incorporated into experimental studies of L. fortunei. Finally, the tolerance of L. fortunei specimens to the stressor tested herein can help us understand the invasive capacity of this mussel during the establishment process.

Resumo A criação bem sucedida de animais em condições de laboratório para processos comerciais ou experimentais é uma cadeia complexa que inclui vários fatores de estresse (ex. coleta e transporte) que tem como consequência a redução das condições naturais do animal, prejuízos econômicos e resultados biológicos inconsistentes. Desde a invasão do bivalve Limnoperna fortunei (Dunker, 1857) na América do Sul, vários estudos têm sido realizados para ajudar no controle e gestão dessa praga em plantas industriais que utilizam água. Relativamente pouca atenção tem sido dada ao processo de criação de L. fortunei em laboratório, sua condição quando exposta ao estresse e sua aclimatação a condições de laboratório. Considerando estes aspectos, os objetivos deste estudo foram: (i) investigar as respostas fisiológicas de L. fortunei submetidos ao processo de depuração e subsequente transporte (sem água/condição seca) em duas temperaturas, analisando as diferentes concentrações de glicogênio e (ii) monitorar as concentrações de glicogênio nos diferentes grupos, quando mantidos por 28 dias em condições de laboratório. Com base nos resultados obtidos, a depuração não afetou nenhum grupo quando eles foram submetidos a oito horas de transporte. A variação da concentração de glicogênio entre os espécimes do campo quando depurados e não depurados, foi significativa apenas em relação à primeira semana em laboratório para o grupo não depurado e à segunda semana para o grupo depurado. Além disto, a temperatura testada não afetou os grupos submetidos ao transporte. As concentrações de glicogénio foram semelhantes as dos espécimes do campo a partir da terceira semana, o que sugere que os espécimes estão aclimatados às condições de laboratoriais neste período de tempo. Assim, os resultados indicam que o transporte ao ar e o tempo de aclimatação podem ser incorporados com sucesso aos estudos experimentais com L. fortunei. Finalmente, o conhecimento sobre a tolerância de L. fortunei ao estresse pode ajudar a entender a capacidade invasiva deste durante o processo de estabelecimento.

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RESUMEN

Successful animal rearing under laboratory conditions for commercial processes or laboratory experiments is a complex chain that includes several stressors (e.g., sampling and transport) and incurs, as a consequence, the reduction of natural animal conditions, economic losses and inconsistent and unreliable biological results. Since the invasion of the bivalve Limnoperna fortunei (Dunker, 1857) in South America, several studies have been performed to help control and manage this fouling pest in industrial plants that use raw water. Relatively little attention has been given to the laboratory rearing procedure of L. fortunei, its condition when exposed to a stressor or its acclimation into laboratory conditions. Considering this issue, the aims of this study are to (i) investigate L. fortunei physiological responses when submitted to the depuration process and subsequent air transport (without water/dry condition) at two temperatures, based on glycogen concentrations, and (ii) monitor the glycogen concentrations in different groups when maintained for 28 days under laboratory conditions. Based on the obtained results, depuration did not affect either of the groups when they were submitted to approximately eight hours of transport. The variation in glycogen concentration among the specimens that were obtained from the field under depurated and non-depurated conditions was significant only in the first week of laboratory growth for the non-depurated group and in the second week for the depurated group. In addition, the tested temperature did not affect either of the groups that were submitted to transport. The glycogen concentrations were similar to those of the specimens that were obtained from the field in third week, which suggests that the specimens acclimated to laboratory conditions during this period of time. Thus, the results indicate that the air transport and acclimation time can be successfully incorporated into experimental studies of L. fortunei[...](AU)

A criação bem sucedida de animais em condições de laboratório para processos comerciais ou experimentais é uma cadeia complexa que inclui vários fatores de estresse (ex. coleta e transporte) que tem como consequência a redução das condições naturais do animal, prejuízos econômicos e resultados biológicos inconsistentes. Desde a invasão do bivalve Limnoperna fortunei (Dunker, 1857) na América do Sul, vários estudos têm sido realizados para ajudar no controle e gestão dessa praga em plantas industriais que utilizam água. Relativamente pouca atenção tem sido dada ao processo de criação de L. fortunei em laboratório, sua condição quando exposta ao estresse e sua aclimatação a condições de laboratório. Considerando estes aspectos, os objetivos deste estudo foram: (i) investigar as respostas fisiológicas de L. fortunei submetidos ao processo de depuração e subsequente transporte (sem água/condição seca) em duas temperaturas, analisando as diferentes concentrações de glicogênio e (ii) monitorar as concentrações de glicogênio nos diferentes grupos, quando mantidos por 28 dias em condições de laboratório. Com base nos resultados obtidos, a depuração não afetou nenhum grupo quando eles foram submetidos a oito horas de transporte. A variação da concentração de glicogênio entre os espécimes do campo quando depurados e não depurados, foi significativa apenas em relação à primeira semana em laboratório para o grupo não depurado e à segunda semana para o grupo depurado. Além disto, a temperatura testada não afetou os grupos submetidos ao transporte. As concentrações de glicogénio foram semelhantes as dos espécimes do campo a partir da terceira semana, o que sugere que os espécimes estão aclimatados às condições de laboratoriais neste período de tempo. Assim, os resultados indicam que o transporte ao ar e o tempo de aclimatação podem ser incorporados com sucesso aos estudos experimentais com L. fortunei[...](AU)

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RESUMEN

Successful animal rearing under laboratory conditions for commercial processes or laboratory experiments is a complex chain that includes several stressors (e.g., sampling and transport) and incurs, as a consequence, the reduction of natural animal conditions, economic losses and inconsistent and unreliable biological results. Since the invasion of the bivalve Limnoperna fortunei (Dunker, 1857) in South America, several studies have been performed to help control and manage this fouling pest in industrial plants that use raw water. Relatively little attention has been given to the laboratory rearing procedure of L. fortunei, its condition when exposed to a stressor or its acclimation into laboratory conditions. Considering this issue, the aims of this study are to (i) investigate L. fortunei physiological responses when submitted to the depuration process and subsequent air transport (without water/dry condition) at two temperatures, based on glycogen concentrations, and (ii) monitor the glycogen concentrations in different groups when maintained for 28 days under laboratory conditions. Based on the obtained results, depuration did not affect either of the groups when they were submitted to approximately eight hours of transport. The variation in glycogen concentration among the specimens that were obtained from the field under depurated and non-depurated conditions was significant only in the first week of laboratory growth for the non-depurated group and in the second week for the depurated group. In addition, the tested temperature did not affect either of the groups that were submitted to transport. The glycogen concentrations were similar to those of the specimens that were obtained from the field in third week, which suggests that the specimens acclimated to laboratory conditions during this period of time. Thus, the results indicate that the air transport and acclimation time can be successfully incorporated into experimental studies of L. fortunei. Finally, the tolerance of L. fortunei specimens to the stressor tested herein can help us understand the invasive capacity of this mussel during the establishment process.

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The susceptibility and suitability of Omalonyx matheroni as an intermediate host of Angiostrongylus vasorum and the characteristics of larval recovery and development were investigated. Mollusks were infected, and from the 3rd to the 25th day after infection, larvae were recovered from groups of 50 individuals. The first observation of L2 was on the 5th day, and the first observation of L3 was on the 10th day. From the 22nd day on, all larvae were at the L3 stadium. Larval recovery varied from 78.2% to 95.2%. We found larval development to be faster in O. matheroni than in Biomphalaria glabrata. Our findings indicate that this mollusk is highly susceptible to A. vasorum. Infective L3 were orally inoculated into a dog, and the prepatent period was 39 days. This is the first study to focus on O. matheroni as an intermediate host of A. vasorum.

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Transplantation of the haematopoietic organ from Biomphalaria tenagophila (Taim strain, RS, Brazil), resistant to Schistosoma mansoni, to a highly susceptible strain (Cabo Frio, RJ, Brazil) of the same species, showed in the recipient snails resistance against the trematode, when a successful transplant occurred. The success of transplantation could be confirmed by a typical molecular marker of the Taim strain in haemocytes of the recipients (350 bp detected by PCR-RFLP). The recipient snails which did not present the donor marker in haemocytes (unsuccessful transplantation) were infected with the parasite. The use of an atoxic modelling clay for closing the hole in the transplantation site reduced significantly the mortality caused by bleeding after transplantation procedures.

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Due to difficulties concerning morphological identification of planorbid snails of the genus Biomphalaria, and given a high variation of characters and in the organs with muscular tissue, we designed specific polymerase chain reaction (PCR) primers for Brazilian snail hosts of Schistosoma mansoni from available sequences of internal transcribed spacer 2 (ITS2) of the ribosomal RNA gene. From the previous sequencing of the ITS2 region, one primer was designed to anchor in the 5.8S conserved region and three other species-specific primers in the 28S region, flanking the ITS2 region. These four primers were simultaneously used in the same reaction (Multiplex-PCR), under high stringency conditions. Amplification of the ITS2 region of Biomphalaria snails produced distinct profiles (between 280 and 350 bp) for B. glabrata, B. tenagophila and B. straminea. The present study demonstrates that Multiplex-PCR of ITS2-DNAr showed to be a promising auxiliary tool for the morphological identification of Biomphalaria snails, the intermediate hosts of S. mansoni.

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Due to difficulties concerning morphological identification of planorbid snails of the genus Biomphalaria, and given a high variation of characters and in the organs with muscular tissue, we designed specific polymerase chain reaction (PCR) primers for Brazilian snail hosts of Schistosoma mansoni from available sequences of internal transcribed spacer 2 (ITS2) of the ribosomal RNA gene. From the previous sequencing of the ITS2 region, one primer was designed to anchor in the 5.8S conserved region and three other species-specific primers in the 28S region, flanking the ITS2 region. These four primers were simultaneously used in the same reaction (Multiplex-PCR), under high stringency conditions. Amplification of the ITS2 region of Biomphalaria snails produced distinct profiles (between 280 and 350 bp) for B. glabrata, B. tenagophila and B. straminea. The present study demonstrates that Multiplex-PCR of ITS2-DNAr showed to be a promising auxiliary tool for the morphological identification of Biomphalaria snails, the intermediate hosts of S. mansoni

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