On the 13C/12C isotopic signal of day and night respiration at the mesocosm level

While there is currently intense effort to examine the ¹³C signal of CO₂ evolved in the dark, less is known on the isotope composition of day‐respired CO₂. This lack of knowledge stems from technical difficulties to measure the pure respiratory isotopic signal: day respiration is mixed up with photorespiration, and there is no obvious way to separate photosynthetic fractionation (pure c_i/c_a effect) from respiratory effect (production of CO₂ with a different δ¹³C value from that of net‐fixed CO₂) at the ecosystem level. Here, we took advantage of new simple equations, and applied them to sunflower canopies grown under low and high [CO₂]. We show that whole mesocosm‐respired CO₂ is slightly ¹³C depleted in the light at the mesocosm level (by 0.2–0.8‰), while it is slightly ¹³C enriched in darkness (by 1.5–3.2‰). The turnover of the respiratory carbon pool after labelling appears similar in the light and in the dark, and accordingly, a hierarchical clustering analysis shows a close correlation between the ¹³C abundance in day‐ and night‐evolved CO₂. We conclude that the carbon source for respiration is similar in the dark and in the light, but the metabolic pathways associated with CO₂ production may change, thereby explaining the different ¹²C/¹³C respiratory fractionations in the light and in the dark.     

Evaluation of Camera Trapping for Estimating Red Fox Abundance

ABSTRACT The nature reserve Serra da Malcata, Portugal, was recently considered a site for Iberian lynx (Lynx pardinus) reintroduction. Because of potential disease risk posed by red foxes (Vulpes vulpes) in the area, a reliable estimate of fox abundance was critical for a dependable reintroduction program. We adapted camera-trapping techniques for estimating red fox abundance in the reserve. From July 2005 to August 2007, we conducted 7 camera-trapping sessions, allowing for individual identification of foxes by physical characteristics. We estimated abundance using the heterogeneity (M_h) model of the software program CAPTURE. Estimated density ranged from 0.91 ± 0.12 foxes/km²to 0.74 ± 0.02 foxes/km². By estimating red fox density, it is possible to define the number of foxes that must be sampled to assess the presence of potential fox-transmitted diseases that may affect lynx reintroduction.     

A review of the genus Semele (Ruscaceae) systematics in Madeira

The present study attempts to review the systematics of Semele (Ruscaceae) in Madeira, based on phenotypic diversity. The variation in some vegetative (climbing shoot, second-order branches or 'phylloclades') and sexual (inflorescence and flowers) characters was analysed in 115 plant specimens from 30 field populations, herbaria of the Costa collection and Madeira Botanical Garden (MADJ) and certain gardens. Thirty-one quantitative and qualitative characters have been utilized in the analysis. Kaiser–Meyer–Olkin (KMO) analysis indicates adequate sampling. Principal component analysis (PCA) reveals that the spatial distribution of individuals has a discontinuous behaviour. Principal coordinate analysis (PCO) utilizing the Gower coefficient on standardized data revealed a significantly discontinuous distribution of individuals, such that two different clusters can be defined. The Student's t -test and Tukey test on separate characters, when individuals were classified according to the Costa classification, confirms the significant differences between grouping accessions. This leads to the recognition of two species within the genus in Madeira. Literature and herbarium studies show that these two taxa are conspecific with Semele androgyna (L.) Kunth sensu stricto (s.s.) and Semele menezesi Costa sensu lato (s.l.) . A separated statistical analysis of the S. androgyna cluster shows the possible existence of additional subgroups. Based on field population distribution, ecological behaviour and variation in features, we propose the recognition of two species, S. androgyna (L.) Kunth and S. menezesi (Costa) Pinheiro de Carvalho, and two subspecies S. androgyna (L.) Kunth androgyna Pinheiro de Carvalho and S. androgyna (L.) Kunth pterygophora Pinheiro de Carvalho.  © 2004 The Linnean Society of London, Botanical Journal of the Linnean Society , 2004, 146 , 483–497.     

Determinants of assemblage structure in Neotropical dry forest lizards

We investigated the structure of a lizard assemblage from Seasonally Dry Tropical Forest enclaves in the Brazilian Cerrado biome, by testing the roles of ecological and historical components. We analysed data from 469 individuals, belonging to 18 lizard species, sampled by a combination of pitfall, funnel and glue traps, as well as by haphazard sampling. Null model analyses and Canonical Phylogenetic Ordination analysis, coupled with Monte Carlo simulations, revealed lack of both ecological and phylogenetic structure in microhabitat use. Conversely, these analyses revealed a mean overlap in diet composition significantly smaller than expected by chance and significant historical structure. Structure in diet composition was due to phylogenetic effects corresponding to the most basal divergence of the squamate phylogeny (Iguania/Scleroglossa) and the clades Teiidae and Gymnophthalmidae. Among lizards, evolutionary constraints on microhabitat use appear less than on prey use, suggesting that the availability of historically preferred prey types moderates microhabitat selection. The lack of structure in microhabitat use suggests absence of competitive interactions on the spatial component. On the other hand, food preferences have a deep historical basis and do not reflect current competitive interactions.     

Use of Camera-Trapping to Estimate Puma Density and Influencing Factors in Central Brazil

Abstract: We used remotely triggered cameras to collect data on Puma (Puma concolor) abundance and occupancy in an area of tropical forest in Brazil where the species' status is poorly known. To evaluate factors influencing puma occupancy we used data from 5 sampling campaigns in 3 consecutive years (2005 to 2007) and 2 seasons (wet and dry), at a state park and a private forest reserve. We estimated puma numbers and density for the 2007 sampling data by developing a standardized individual identification method. We based individual identification on 1) time-stable parameters (SP; physical features that do not change over time), and 2) time-variable parameters (VP; marks that could change over time such as scars and botfly marks). Following individual identification we established a capture-recapture history and analyzed it using closed population capture-mark-recapture models. Puma capture probability was influenced by camera placement (roads vs. trails), sampling year, and prey richness. Puma occupancy was positively associated with species richness and there was a correlation between relative puma and jaguar (Panthera onca) abundance. Identifications enabled us to generate 8 VP histories for each photographed flank, corresponding to 8 individuals. We estimated the sampled population at 9 pumas (SE = 1.03, 95% CI = 8–10 individuals) translating to a density of 3.40 pumas/100 km². Information collected using camera-traps can effectively be used to assess puma population size in tropical forests. As habitat progressively disappears and South American felines become more vulnerable, our results support the critical importance of private forest reserves for conservation.     

Life history of the Mato Grosso Antbird Cercomacra melanaria in the Brazilian Pantanal

Here we describe the habitat use, reproductive biology, diet and details of the life history of the Mato Grosso Antbird Cercomacra melanaria , one of the least studied Cercomacra species. We located 30 nests, of which 22 became active and were monitored. The nest consists of a deep cup, placed among dense tangles of vegetation, almost always in forests of Vochysia divergens ( Cambarazal ). Eggs are pinkish-cream coloured, covered with reddish brown to purple spots and blotches that are concentrated around its centre and larger end. Average nest success was 32%. The Mayfield method indicated daily survival rates during egg and nestling periods of 0.94 and 0.96, respectively, and a success rate of 26%. Examination of 24 stomach contents revealed a diet based exclusively on arthropods.     

Comparing patterns of nuclear and mitochondrial divergence in a cryptic species complex: the case of Iberian and North African wall lizards (Podarcis, Lacertidae)

Combining different sources of information is essential for a complete understanding of the process of genetic differentiation between species. The Iberian and North African wall lizard ( Podarcis ) species complex has been the object of several studies regarding morphological and mitochondrial DNA variation but, so far, no large-scale survey of nuclear variation within this group has been accomplished. In this study, ten polymorphic allozyme loci were studied in 569 individuals collected across the Iberian Peninsula and North Africa. The obtained data were analysed using both conventional population genetic tools and recent Bayesian model-based clustering methods. Our results show that there are several well-differentiated entities corroborating the major splits observed in mtDNA analyses. These groups correspond not only to the fully recognized species Podarcis bocagei , Podarcis carbonelli , and Podarcis vaucheri but also to multiple forms within the polytypic Podarcis hispanica , all of which have a similar level of differentiation to that observed between the acknowledged species. However, relationships between forms are weakly supported both by population and individual clustering methods, suggesting a scenario of a rapid diversification that contrasts to the clear bifurcating model assumed from previous mtDNA analyses. Individual multilocus analyses report few individuals misassigned or apparently admixed, some of which are most likely explained by the persistence of high levels of ancestral polymorphism. Other admixed individuals, however, are probably the result of limited levels of gene flow between forms.  © 2007 The Linnean Society of London, Biological Journal of the Linnean Society , 2007, 91 , 121–133.     

Recombinant expression and characterization of a lysozyme from the midgut of Triatoma brasiliensis (Hemiptera,Reduviidae) in comparison with intestinal muramidase activity

Insect c‐type lysozymes are antibacterial proteins that are synthesized in different organs with high activity against Gram‐positive bacteria. Because lysozymes possess muramidase activity, they also play an important role in the digestion of bacteria in Diptera. Triatomines express lysozyme‐encoding genes constitutively in the anterior region (cardia and stomach) of the midgut and the fat body after injection of bacteria into the haemocoel. The present study describes the overexpression of the Triatoma brasiliensis lysozyme 1 (lys1) in Escherichia coli. Recombinant T. brasiliensis Lys1 (TbLys1) is purified after solubilization of the inclusion bodies. The protein refolds successfully, showing muramidase activity against Micrococcus lysodeikticus lyophilized cells, after enterokinase cleavage of its thioredoxin fusion protein. In in‐gel zymograms and turbidimetric liquid assays TbLys1 is broadly active under alkaline and acid conditions, indicating a possible digestive function in the two physiologically different midgut regions of the bug: the stomach and small intestine. Muramidase activity is shown in the stomach and small intestine content of unfed bugs and bugs at different days after feeding, respectively. Western blot analysis identifies TbLys1 as lysozyme.     

A three‐way contact zone between forms of Patella rustica (Mollusca: Patellidae) in the central Mediterranean Sea

Previous studies have reported the occurrence of three differentiated mtDNA lineages within Patella rustica in the Mediterranean Sea. Two hypotheses have been proposed to explain these observations: (1) the maintenance of ancestral polymorphism within a single species; (2) the occurrence of cryptic species not identified previously. To distinguish between these hypotheses, we screened the genetic variability at nine allozyme loci, an intron from the α‐amylase gene and a mitochondrial gene for 187 individuals of P. rustica sampled from seven Mediterranean localities. Eight additional localities were screened for the last two markers to place the differentiated lineages in a clear geographic context. Our results demonstrate that the three mtDNA lineages correspond to three distinct nuclear genotype clusters and provide further details on their distribution: the cluster corresponding to the mtDNA lineage from the Atlantic and western Mediterranean extends as far as the south coast of Italy, whereas the remaining two clusters occur in sympatry in the eastern Mediterranean. One of the eastern Mediterranean clusters is highly differentiated and seems to be reproductively isolated from the codistributed form; we therefore suggest that it corresponds to a new species. The remaining two clusters are less differentiated and form a contact zone across south Italian shores. This three‐way contact zone constitutes an interesting model for the study of speciation in the marine realm. © 2010 The Linnean Society of London, Biological Journal of the Linnean Society, 2010, 100 , 154–169.     

Molecular detection of Wolbachia pipientis in natural populations of mosquito vectors of Dirofilaria immitis from continental Portugal: first detection in Culex theileri

Wolbachia pipientis (Rickettsiales: Rickettsiaceae) protects mosquitoes from infections with arboviruses and parasites. However, the effect of its co‐infection on vector competence for Dirofilaria immitis (Spirurida: Onchocercidae) in the wild has not been investigated. This study aimed to screen vectors of D. immitis for wPip, to characterize these, and to investigate a possible association between the occurrence of W. pipientis and that of the nematode. The presence of W. pipientis was assessed in the five mosquito potential vectors of D. immitis in Portugal. Polymerase chain reaction (PCR) products were sequenced, and wPip haplotypes were determined by PCR–restricted fragment length polymorphism (RFLP). Results showed that wPip was detected in 61.5% of Culex pipiens (Diptera: Culicidae) pools and 6.3% of Culex theileri pools. wPip 16s rRNA sequences found in Cx. theileri exactly match those from Cx. pipiens, confirming a mosquito origin, rather than a nematode origin, as some specimens were infected with D. immitis. Only wPip haplotype I was found. No association was found between the presence of wPip and D. immitis in mosquitoes and hence a role for this endosymbiont in influencing vectorial competence is yet to be identified. This study contributes to understanding of wPip distribution in mosquito populations and, to the best of the authors' knowledge, is the first report of natural infections by wPip in Cx. theileri.