Geographic differentiation of <Emphasis Type="Italic">Cobitis shikokuensis</Emphasis> inferred from mtDNA RFLP analysis

Mitochondrial DNA divergence among populations of the Japanese spinous loach Cobitis shikokuensis, endemic to Shikoku Island, was investigated by restricted fragment length polymorphism analysis. A total of 68 restriction sites on DNA fragments from the cytochrome b to D-loop regions and from the 12S rRNA to 16S rRNA regions, amplified by PCR, were analyzed. A total of 12 haplotypes (plus 6 in outgroups) were detected in 268 specimens collected from 19 localities in seven rivers (and 41 specimens from four localities in three rivers in outgroups). Three of the seven river populations of C. shikokuensis were shown to have unique haplotypes, and four of the seven river populations were monomorphic. The nested structure of the haplotype network for populations of C. shikokuensis exhibited two large clades corresponding to (1) populations from the Shimanto River and its neighbors and (2) two genetically divergent populations in the Shigenobu and Iwamatsu Rivers. The population from the Shimanto River, the largest river inhabited by C. shikokuensis, maintains great haplotype diversity as well as the allozyme diversity previously reported. On the other hand, populations from the Hiji River, the second largest river, which exhibited the highest allozyme diversity, were monomorphic in their mtDNA. The nested clade analysis (NCA) revealed that past fragmentation between the above two clades could occur in the initial distribution process of C. shikokuensis. The large genetic divergence of two river populations from the Shigenobu and Iwamatsu Rivers was inferred to be caused by a process of long distance colonization and fragmentation. MtDNA introgression into the Hiji River population from southern river populations was suggested. Taking genetic divergence into consideration, each river population of C. shikokuensis should be conserved separately as like a distinct species, and conservation programs for the small populations showing less genetic variability should be invoked as soon as possible.     

Two New Genera of Baetidae (Ephemeroptera; Insecta) from Madagascar

Two new genera of Baetidae (Ephemeroptera), Scutoptilum gen. n. and Echinopus gen. n., are described to accommodate three new species from Madagascar: Scutoptilum verrucosum sp. n., Echinopus giboni sp. n., Echinopus minutus sp. n.. They all share a unique apomorphy; they possess a subapico-transverse arc of long setae on the outer margin of femora. Scutoptilum presents important adaptations to fast flowing waters and mouthparts modified for scraping. Moreover, Scutoptilum shows an unusual general habitus with a prothorax broadly expanded laterally, forewing pads extremely developed and shape of the head subrectangular. Echinopus appears less derived and possesses more plesiomorphic features. Both genera are closely related to the Afrotropical genus Afroptilum and belong to the Centroptiloides complex.     

Observations on the life stages of Sphaerothecum destruens n. g., n. sp., a mesomycetozoean fish pathogen formerly referred to as the rosette agent [correction

The rosette agent is an obligate intracellular parasite that causes morbidity and mortality in salmonid fish. In laboratory cultures, the spore stage (2-6 microm diam.) replicates in a salmonid cell line by sequential asexual division, giving rise to daughter cells. If infected cell cultures are transferred to distilled water, the spore stage undergoes internal division to give rise to at least 5 cells each of which develops into a uniflagellated zoospore with a body of approximately 2 microm and a flagellum approximately 10 microm long. Zoosporulation does not occur in cell culture medium alone, artificial seawater, or phosphate-buffered saline. This parasite is currently classified as a member of the Class Mesomycetozoea (formerly Ichthyosporea) based on phylogenetic analyses of the small subunit ribosomal DNA of three different isolates from fish. Given these new morphological observations combined with the available molecular phylogenetic data on other mesomycetozoeans, we propose to classify the rosette agent as Sphaerothecum destruens, n. g., n. sp. This new genus has unique features including (1) intracellular development of spore stages in various organs eliciting a host granulomatous response; and (2) the differentiation of mature spores into multiple, flagellated zoospores. Taken together, these characteristics clearly distinguish it from the closely related genera Dermocystidium and Rhinosporidium.     

Phylogenetic Analysis of the Hoplolaiminae Inferred from Combined D2 and D3 Expansion Segments of 28S rDNA

DNA sequences of the D2-D3 expansion segments of the 28S gene of ribosomal DNA from 23 taxa of the subfamily Hoplolaiminae were obtained and aligned to infer phylogenetic relationships. The D2 and D3 expansion regions are G-C rich (59.2%), with up to 20.7% genetic divergence between Scutellonema brachyurum and Hoplolaimus concaudajuvencus. Molecular phylogenetic analysis using maximum likelihood and maximum parsimony was conducted using the D2-D3 sequence data. Of 558 characters, 254 characters (45.5%) were variable and 198 characters (35.4%) were parsimony informative. All phylogenetic methods produced a similar topology with two distinct clades: One clade consists of all Hoplolaimus species while the other clade consists of the rest of the studied Hoplolaiminae genera. This result suggests that Hoplolaimus is monophyletic. Another clade consisted of Aorolaimus, Helicotylenchus, Rotylenchus, and Scutellonema species. Phylogenetic analysis using the outgroup species Globodera rostocheinsis suggests that Hoplolaiminae is paraphyletic. In this study, the D2-D3 region had levels of DNA sequence divergence sufficient for phylogenetic analysis and delimiting species of Hoplolaiminae.     

A work‐for‐food enrichment program increases exploration and decreases stereotypies in four species of bears

Zoo‐housed bears are prone to exhibiting stereotypic behaviors, generally considered indicators of negative welfare. We explored the effects of a variable‐time feeding enrichment schedule on behavioral indicators of welfare in four bear species at Cleveland Metroparks Zoo. We distributed the diets of eight bears in one of five enrichment items, for two consecutive days each, and monitored behavior throughout the day. In Experiment 1, we compared variable‐time to fixed‐time presentation of enrichment over two, 10‐day periods. Overall, bears performed more exploratory behavior when enriched (p < 0.0001). Furthermore, variable‐time enrichment was associated with a greater increase in exploratory behavior than fixed‐time enrichment when compared to baseline (p < 0.001). Both fixed‐time (p_unadjusted <0.05, p_adjusted = 0.07) and variable‐schedule (p_unadjusted <0.05, p_adjusted = 0.09) enrichment were also associated with similar decreases in abnormal behavior compared to baseline. For Experiment 2, we tested habituation to enrichment over 30 days using multiple items and a semi‐variable presentation schedule. Again during the enrichment period, bears exhibited increased exploratory behavior (p < 0.0001) and decreased abnormal behaviors compared to baseline (p_unadjusted = 0.05, p_adjusted = 0.09). We observed no habituation during the 30‐day sustained enrichment period for these behaviors. Collectively, these results suggest that daily, variable‐schedule feeding enrichment, with intermittent presentation of unique enrichment items, increases behavioral indicators of positive welfare and decreases behavioral indicators of negative welfare.     

Genome sequence of Phaeobacter daeponensis type strain (DSM 23529T), a facultatively anaerobic bacterium isolated from marine sediment,and emendation of Phaeobacter daeponensis

TF-218^T is the type strain of the species Phaeobacter daeponensis Yoon et al. 2007, a facultatively anaerobic Phaeobacter species isolated from tidal flats. Here we describe the draft genome sequence and annotation of this bacterium together with previously unreported aspects of its phenotype. We analyzed the genome for genes involved in secondary metabolite production and its anaerobic lifestyle, which have also been described for its closest relative Phaeobacter caeruleus. The 4,642,596 bp long genome of strain TF-218^T contains 4,310 protein-coding genes and 78 RNA genes including four rRNA operons and consists of five replicons: one chromosome and four extrachromosomal elements with sizes of 276 kb, 174 kb, 117 kb and 90 kb. Genome analysis showed that TF-218^T possesses all of the genes for indigoidine biosynthesis, and on specific media the strain showed a blue pigmentation. We also found genes for dissimilatory nitrate reduction, gene-transfer agents, NRPS/ PKS genes and signaling systems homologous to the LuxR/I system.     

Estudio Fenológico de Cactíceas en el Enclave Seco de la Tatacoa,Colombia1

A one‐year phenological study of three columnar cacti, Stenocereus griseus (Haw.) Britton & Rose, Pilosocereus sp., Cereus hexagonus (L.) Mill., and a decumbent cactus Monvillea cf. smithiana (Britton & Rose) Backeberg., was carried out in the Andean arid region of La Tatacoa, Colombia. Pollinators and/or dispersers of the cacti species also were studied monthly, and fecal samples were collected for the identification of pollen and seeds. The flowering of all species was prolonged and showed bimodal, multimodal, or irregular patterns. Fruiting in all species also was prolonged and followed flowering with a lag of less than two months. Although there were no simple correlations between rainfall and flowering or fruiting, flower production during the dry season was higher for 5. griseus, while Pilosocereus sp. and C. hexagonus showed higher flower production during the wet season. Fruit production was also seasonal, with higher production during the wet season for 5. griseus and C. hexagonus. The patterns of flowering and fruiting in M. cf. smithiana showed no relationships with dry and wet seasons. The bats Glossophaga longirostris, Carollia perspicillata, Sturnira lilium, the birds Melanerpes rubricapillus (Picidae) and Mimus gilvus (Mimidae), and moths of the family Sphingidae, were identified as pollinators and/or fruit consumers of these cacti species.     

Phylogeography of the high alpine plant Senecio halleri (Asteraceae) in the European Alps: in situ glacial survival with postglacial stepwise dispersal into peripheral areas

Whether alpine plant species survived Pleistocene glaciations in situ on high alpine nunatak mountains is still under debate. To test this hypothesis, Senecio halleri, a high alpine and endemic species with a narrow distribution range in the European Alps, was chosen as a model organism. Polymerase chain reaction-restriction fragment length polymorphisms of chloroplast DNA (cpDNA PCR-RFLPs) were used in a phylogeographic analysis of 14 populations of S. halleri, covering its total distribution area. The results of haplotype diversity and distribution gave evidence of in situ glacial survival on siliceous central-alpine nunatak mountains in two areas, southwest and northeast of the Aosta valley. According to the absence of genetic differentiation between these two nunatak areas (based on amova), nested clade analysis implied a history of preglacial gene flow, in situ survival and extinction of intermediate populations during glaciation and postglacial stepwise recolonization of peripheral and intermediate areas.     

Comparison of lead contamination among related scavenging birds sharing food sources: a call for caution when using surrogate species

Lead contamination is a global problem affecting a large number of bird species around the world. Among the different avian guilds, vultures and facultative scavengers are particularly threatened by this toxic metal. However, little information is available about differences in exposure to this metal for sympatric vulture species that share food resources. We compared blood lead concentrations of two closely related sympatric obligate scavenger species, the abundant Black Vulture Coragyps atratus and the threatened Andean Condor Vultur gryphus in north-western Patagonia, Argentina. We sampled 28 Andean Condors and 29 Black Vultures trapped foraging in the same area in the steppe. We also sampled 16 Black Vultures foraging in a rubbish dump to determine whether there were differences in lead contamination among foraging sites. Andean Condors had significantly higher mean blood lead concentrations than Black Vultures. There was no difference in lead concentrations between Black Vultures trapped in the steppe and in the rubbish dump. The prevalence and probability of lead concentrations above the threshold level (20 µg/dL) was higher for Andean Condors than for Black Vultures, potentially producing different effects on their health. This disparity in lead contamination may be due to differences in their foraging habits or in their susceptibility to this toxic metal. Overall, our results suggest caution in using an abundant surrogate species to infer lead contamination in a closely related but harder to sample species.