The first fossil Paussine (Coleoptera: Carabidae) from Mexican amber

Eohomopterus simojovelensis n. sp., the first fossil record of the subfamily Paussinae (Coleoptera: Carabidae) from the Miocene amber of the Simojovel area, Chiapas, Mexico, is described. The morphology of the new species is compared withEohomopterus poinari Nagel, 1997 from Dominican amber as well as with extant representatives ofEohomopterus, and the biogeographical implications are discussed.      

Bacillus DNA in fossil bees: an ancient symbiosis?

We report here the isolation of DNA from abdominal tissue of four extinct stingless bees (Proplebeia dominicana) in Dominican amber, PCR amplification of a 546-bp fragment of the 16S rRNA gene from Bacillus spp., and their corresponding nucleotide sequences. These sequences were used in basic local alignment search tool searches of nonredundant nucleic acid data bases, and the highest scores were obtained with 16S rRNA sequences from Bacillus spp. Phylogenetic inference analysis by the maximum-likelihood method revealed close phylogenetic relationships of the four presumed ancient Bacillus sequences with Bacillus pumilus, B. firmus, B. subtilis, and B. circulans. These four extant Bacillus spp. are commonly isolated from abdominal tissue of stingless bees. The close phylogenetic association of the extracted DNA sequences with these bee colonizers suggests that a similar bee-Bacillus association existed in the extinct species P. dominicana.     

How long can insect species exist? Evidence from extant and fossil Micromalthus beetles (Insecta: Coleoptera)

Six new fossils of Micromalthus (Coleoptera: Archostemata) from Dominican amber are compared with extant and previously described fossil micromalthid beetles. The amber inclusions are well preserved and all important morphological features are visible. Taking into account the morphological variability of the extant species Micromalthus debilis LeConte, 1878 , it is not possible to find any morphological features that distinguish the fossils from the extant species. This also applies to the Dominican amber inclusion described as Micromalthus anasi Perkovsky, 2008, which therefore is considered a junior synonym of M. debilis. The lack of morphological change in M. debilis over time might possibly be explained by unusually stable environmental conditions, as this species occupies a very specialized ecological niche in decaying timber. A general survey of fossil insects indistinguishable from extant species is presented. © 2009 The Linnean Society of London, Zoological Journal of the Linnean Society, 2010, 158 , 300–311.     

First fossil Molinaranea Mello‐Leitão, 1940 (Araneae: Araneidae), from middle Miocene Dominican amber,with a phylogenetic and palaeobiogeographical analysis of the genus

The first fossil Molinaranea is described, from middle Miocene Dominican amber. This record extends the known range of the genus back 16 million years; it also extends the geographical range of the genus through time, with extant species known only from Chile, Argentina, the Falkland Islands, and Juan Fernandez Island. A parsimony‐based phylogenetic analysis was performed, which indicates that the fossil species, Molinaranea mitnickii sp. nov. , is nested with Molinaranea magellanica Walckenaer, 1847 and Molinaranea clymene Nicolet, 1849 . A modified Brooks parsimony analysis was conducted in order to examine the biogeography and origins of the fossil species in the Dominican Republic; the analysis suggests that M. mitnickii sp. nov. arrived in Hispaniola from South America as a result of a chance dispersal event. © 2010 The Linnean Society of London, Zoological Journal of the Linnean Society, 2010, 158 , 711–725.     

New Tridactyloidea in Miocene amber from the Dominican Republic (Orthoptera: Caelifera)

Two new Tridactyloidea of the families Ripipterygidae and Tridactylidae are described from the Early Miocene (Burdigalian) amber of the Dominican Republic, namely: Archaeoellipes engeli n. gen., n. sp. (Tridactylidae: Tridactylinae) and Mirhipipteryx antillarum n. sp. (Ripipterygidae). These new taxa represent the fi rst record of Tridactyloidea in Dominican amber and the fi rst fossil record of Ripipterygidae, which are otherwise known only from the extant fauna.     

Identification of Soil Microbes Capable of Utilizing Cellobiosan

Approximately 100 million tons of anhydrosugars, such as levoglucosan and cellobiosan, are produced through biomass burning every year. These sugars are also produced through fast pyrolysis, the controlled thermal depolymerization of biomass. While the microbial pathways associated with levoglucosan utilization have been characterized, there is little known about cellobiosan utilization. Here we describe the isolation and characterization of six cellobiosan-utilizing microbes from soil samples. Each of these organisms is capable of using both cellobiosan and levoglucosan as sole carbon source, though both minimal and rich media cellobiosan supported significantly higher biomass production than levoglucosan. Ribosomal sequencing was used to identify the closest reported match for these organisms: Sphingobacterium multivorum, Acinetobacter oleivorans JC3-1, Enterobacter sp SJZ-6, and Microbacterium sps FXJ8.207 and 203 and a fungal species Cryptococcus sp. The commercially-acquired Enterobacter cloacae DSM 16657 showed growth on levoglucosan and cellobiosan, supporting our isolate identification. Analysis of an existing database of 16S rRNA amplicons from Iowa soil samples confirmed the representation of our five bacterial isolates and four previously-reported levoglucosan-utilizing bacterial isolates in other soil samples and provided insight into their population distributions. Phylogenetic analysis of the 16S rRNA and 18S rRNA of strains previously reported to utilize levoglucosan and our newfound isolates showed that the organisms isolated in this study are distinct from previously described anhydrosugar-utilizing microbial species.     

Paleorhodococcus dominicanus n. gen., n sp. (Actinobacteria) in a faecal droplet of Triatoma dominicana (Hemiptera: Reduviidae: Triatominae) in Dominican amber

Paleorhodococcus dominicanus n. gen., n sp. (Actinobacteria) is described from a faecal droplet of Triatoma dominicana (Hemiptera: Reduviidae: Triatominae) in Dominican amber. The fossil can be distinguished from the species of the closely related extant genus Rhodococcus Zopf 1891 by its spherical–ellipsoidal cocci forming substrate filaments with elementary branching, the clustering of coccoidal elements, the short filaments bearing reduced side branches and its occurrence in a faecal droplet of the extinct triatomine bug, P. dominicanus. This is the first fossil record of an Actinobacter, which shows that these organisms formed symbiotic associations with insects by the mid-Tertiary.     

13C NMR and IR analyses of structure,aging and botanical origin of Dominican and Mexican ambers

¹³C NMR analysis of Oligocene amber from the Dominican Republic, Oligo-Miocene amber from Mexico and trunk resins from certain extant species of Hymenaea, suggests a polylabdane structure for all and gives insights into structural changes during aging. Additionally, IR spectra, some aspects of the ¹³C NMR spectra and paleobotanical data suggest a close relationship between Dominican amber and H. verrucosa. By contrast, spectral evidence shows that Mexican amber differs, not only from the former polymers, but from the H. courbaril polymer.     

A walking stick,Clonistria dominicana n. sp. (Phasmatodea: Diapheromeridae) in Dominican amber

A walking stick, Clonistria dominicana n. sp. (Phasmatodea: Diapheromeridae: Diapheromerinae: Diapheromerini) is described from Dominican amber. The new species can be distinguished from extant members of the genus by the long hind leg tarsomere II (over six times the length of the foreleg tarsomere II), the short metanotum, which is only half the length of the median segment and the prominent median carina. Two phasmatid eggs in separate pieces of Dominican amber are also presented. One of these has characteristics of the genus Clonistria Stål, 1875, which supports the generic placement of C. dominicana. The second egg resembles those of the genus Malacomorpha Rehn, 1906 (Pseudophasmatidae: Pseudophasmatinae). Both of these genera occur in Hispaniola today.     

A scolopocryptopid centipede (Chilopoda: Scolopendromorpha) from Mexican amber: synchrotron microtomography and phylogenetic placement using a combined morphological and molecular data set

The first scolopocryptopid centipede known from the fossil record is a specimen of the subfamily Scolopocryptopinae in Miocene amber from Chiapas, southern Mexico. It is described here as Scolopocryptops simojovelensis sp. nov. , displaying a distinct combination of morphological characters compared to extant congeners. Anatomical details of the fossil specimen were acquired by non‐invasive 3D synchrotron microtomography using X‐ray phase contrast. The phylogenetic position of the new species is inferred based on a combination of morphological data with sequences for six genes (nuclear 18S and 28S rRNA, nuclear protein‐coding histone H3, and mitochondrial 12S rRNA, 16S rRNA, and protein‐coding cytochrome c oxidase subunit I) for extant Scolopendromorpha. The data set includes eight extant species of Scolopocryptops and Dinocryptops from North America, east Asia, and the Pacific, rooted with novel sequence data for other blind scolopendromorphs. The molecular and combined data sets, analysed in a parsimony/direct optimization framework, identified a stable pattern of two main clades within Scolopocryptopinae. North American and Asian species of Scolopocryptops are united as a clade supported by both morphological and molecular characters. Its sister group is a Neotropical clade in which the type species of Dinocryptops is nested within a paraphyletic assemblage of Scolopocryptops species; Dinocryptops is placed in synonymy with Scolopocryptops. The strength of support for the relationships of extant taxa from the molecular data allow the Chiapas fossil to be assigned with precision, despite ambiguity in the morphological data; the fossil is resolved as sister species to the extant Laurasian clade. © 2012 The Linnean Society of London, Zoological Journal of the Linnean Society, 2012, 166 , 768–786.     

Morphological conservatism in the foreleg structure of cicada hatchlings,Burmacicada protera n. gen., n. sp. in Burmese amber,Dominicicada youngi n. gen., n. sp. in Dominican amber and the extant Magicicada septendecim (L.) (Hemiptera: Cicadidae)

Two cicada hatchlings (Hemiptera: Cicadidae) in Burmese and Dominican amber are described as Burmacicada protera n. gen., n. sp. and Dominicicada youngi n. gen., n. sp., respectively. Although very similar in appearance, the two species can be separated by body contour, the nature of the process on the terminal antennomere and the shape and size of protrusions, teeth and spines on the forelegs. A comparison of the forelegs of the fossil hatchlings with those of an extant hatchling of the periodical cicada, Magicicada septendecim (L.), reveals a remarkable degree of morphological conservatism over 100 million years. A brief review of fossil cicadas is presented.     

Detection of Deefgea chitinilytica in freshwater ornamental fish

Aim: To identify and characterize six chitinolytic bacterial strains isolated from ornamental fish. Methods and Results: Six different isolates of Deefgea chitinilytica were detected in healthy as well as diseased ornamental fish in Germany over a period of 2 years. Bacterial strains were identified using 16S rRNA partial gene sequencing and further characterized using different biochemical microtest systems and additional standard biochemical tests. Conclusion: We show that commercially available biochemical microtest systems are useful for identification of D. chitinilytica, supplemented by 16S rRNA partial gene sequencing. Furthermore, this study provides new information about the occurrence of D. chitinilytica, as this is the first isolation of D. chitinilytica from animals and first described isolation in Europe. Significance and Impact of the Study: Deefgea chitinilytica may be isolated regularly in fish diagnostic laboratories. Therefore, accurate identification of this bacterial species is important. Involvement of D. chitinilytica in opportunistic infections of aquatic organisms cannot be excluded and has to be further investigated.     

Two new species of the genus Rhynchitobius Sharp, 1889 (Coleoptera: Rhynchitidae) in Dominican amber

New species of rhynchitid weevils, Rhynchitobius tanyrhinus n. sp. and R. xuthocolus n. sp. (Coleoptera: Rhynchitidae) in the tribe Auletini are described from Dominican amber. The former species is characterized by a long rostrum, narrow pronotum and elytra, rugose elytral intervals and distinct elytral striae. The latter species has a short rostrum, wide antennal club, yellow legs, non-rugose elytral intervals and weak elytral striae. The present study describes the first representatives of the family Rhynchitidae from Dominican amber.     

Dilution-to-Extinction Culturing of Psychrotolerant Planktonic Bacteria from Permanently Ice-covered Lakes in the McMurdo Dry Valleys,Antarctica

Lakes in the McMurdo Dry Valleys of Antarctica are characterized by a permanent ice cover and little or no anthropogenic influence. Although bacterial cultures have been obtained from these habitats, recent culture-independent studies indicate that the most abundant microbes in these systems are not yet cultivated. By using dilution-to-extinction cultivation methods with sterilized and nutrient-amended lake water as media, we isolated 148 chemotrophic psychrotolerant bacterial cultures from fresh surface water of Lake Fryxell and the east lobe of Lake Bonney and the hypersaline, suboxic bottom water from the west lobes of Lake Bonney. Screening of the 16S ribosomal ribonucleic acid (rRNA) genes of the cultures by restriction fragment length polymorphism (RFLP) yielded 57 putatively pure psychrotolerant, slow growing cultures grouped into 18 clusters. The sequencing of 16S rRNA genes of randomly selected representatives of each RFLP cluster revealed that the corresponding isolates belong to the Alphaproteobacteria (six RFLP patterns), Betaproteobacteria (six RFLP patterns), Bacteroidetes (four RFLP patterns), and Actinobacteria (two RFLP patterns). Phylogenetic analysis of the sequences showed that the vast majority of the isolates were not closely related to previously described species. Thirteen of 18 RFLP patterns shared a 16S ribosomal deoxyribonucleic acid sequence similarity of 97% or less with the closest described species, and four isolates had a sequence similarity of 93% or less with the nearest described species. Phylogenetic analysis showed that these sequences were representatives of deeply branching organisms in the respective phylum. A comparison of the isolates with 16S rRNA clone libraries prepared from the same environments showed substantial overlap, indicating that dilution-to-extinction culturing in natural lake water media can help isolate some of the most abundant organisms in these perennially ice-covered lakes.     

Intergeneric Relationships Among Macropodoidea (Metatheria: Diprotodontia) and The Chronicle of Kangaroo Evolution

The superfamily of kangaroos (Macropodoidea) is comprised of the subfamilies Propleopinae, Hypsiprymnodontinae, Paleopotoroinae, Potoroinae, Bulungamayinae, Balbarinae, Macropodinae, and Sthenurinae. Of these, Hypsiprymnodontinae, Potoroinae, and Macropodinae are extant. Competing phylogenetic hypotheses unite potoroines with either hypsiprymnodontines or macropodines, with most recent workers following a classificatory scheme that recognizes Hypsiprymnodontidae (hypsiprymnodontines) and Macropodidae (macropodines + potoroines). To address phylogenetic relationships among living macropodoids, we analyzed sequences from three mitochondrial genes (12S rRNA, tRNA valine, 16S rRNA) and one nuclear gene (protamine P1). MtDNA and protamine P1 both support a basal split of Hypsiprymnodon from other macropodoids rather than an association of Hypsiprymnodon with potoroines. This suggests that bipedal hopping and a complex stomach evolved once among macropodids. Monophyly of the Macropodinae is supported. Among macropodines, there is support for a Dorcopsis-Dorcopsulus association. Potoroine monophyly is less clear, although among potoroines there is support for an association of Bettongia and Aepyprymnus. Divergence times were estimated using 12S rRNA, tRNA-valine, and 16S rRNA transversions and suggest that kangaroos separated from a possum-like ancestor approximately 38–44 million years ago. Hypsiprymnodon diverged from other macropodoids approximately 34 to 38 million years ago. In agreement with the fossil record, the diversification of potoroines predates the diversification of macropodines. The latter have radiated in association with the development of a more arid climate and emergent grasslands over the Australian continent.     

A fungal-like organism associated with a wasp (Hymenoptera: Pteromalidae) in Dominican amber

A fungal-like organism emerging from a parasitic wasp (Hymenoptera: Pteromalidae) in Dominican amber is characterized. The fossil consists of a white sclerotium-like formation in the wasp's abdomen and a flattened clava-like structure with an ovoid terminus emerging from the sclerotium-like formation. The ovoid terminus bears a protruding elliptical appendix. The fossil, which is characterized by its small size, somatic configuration, pteromalid host and presence in Dominican amber, cannot be placed with assurance in any extant fungal group at this time.     

Cloning and characterization of genes coding for ribosomal RNA inCampylobacter jejuni

The DNA fragments coding for ribosomal RNA inCampylobacter jejuni have been cloned from a genomic library ofC. jejuni constructed inEscherichia coli. Clones carrying DNA Sequences for rRNA were identified by hybridization of 5-end-labeled rRNA fromC. jejuni to colony blots of transformants from this gene library. Cloned DNA sequences homologous to each of 5S, 16S, and 23S rRNA were idenfified by hybridization of labeled plasmid DNA to Northern blots of rRNA. The gene coding for 23S rRNA was found to be located on a 5.5kb HindIII fragment, while the 5S and 16S rRNA genes were on HindIII fragments of 1.65 and 1.7 kb, respecitively. The DNA fragment containing the 16S rRNA gene was characterized by restriction endonuclease mapping, and the location of the 16S rRNA gene on this fragment was determined by hybridization of 5-end-labeled rRNA to restriction fragments and also by DNA sequence determination. It appears that the major portion of the coding region for 16S rRNA is located on the 1.7-kb HindIII fragment, while a small portion is carried on an adjacent HindIII fragment of 7.5 kb. Cloned rRNA genes fromC. jejuni were used to study the organization of the rDNA inC. jejuni and other members of the genùsCampylobacter.     

A fossil scorpion Tityus geratus new species (Scorpiones: Buthidae) from Dominican Amber

A fossil scorpion, Tityus geratus (Scorpioncs: Buthidae) n. sp., is described from Dominican amber. This is the third described amber Tityus sp. and the third Dominican amber scorpion. On the basis of the amber finds and available knowledge on the present distribution of Tityus spp. in the Antilles, the vicariance model of Caribbean biogeography can be evoked to best explain the present distribution of this genus.     

First fossil horsefly (Diptera: Tabanidae) in Miocene Mexican amber

The fossil record of the Tabanidae is sparse when compared with other families of Diptera. Even in amber they are rare, probably because of their size and specific flight behavior. Horseflies from amber are only known from Cretaceous age New Jersey amber as well as from the Tertiary age Baltic and Dominican amber, but are herein described for the first time, with Stenotabanus oleariorum sp. n., from Mexican amber. The new species is compared to the fossil horseflies of the same genus S. brodzinskyi Lane, Poinar and Fairchild 1988 and S. woodruffi Lane and Fairchild 1989 from Dominican amber.     

Specific single-cell isolation and genomic amplification of uncultured microorganisms

We in this study describe a new method for genomic studies of individual uncultured prokaryotic organisms, which was used for the isolation and partial genome sequencing of a soil archaeon. The diversity of Archaea in a soil sample was mapped by generating a clone library using group-specific primers in combination with a terminal restriction fragment length polymorphism profile. Intact cells were extracted from the environmental sample, and fluorescent in situ hybridization probing with Cy3-labeled probes designed from the clone library was subsequently used to detect the organisms of interest. Single cells with a bright fluorescent signal were isolated using a micromanipulator and the genome of the single isolated cells served as a template for multiple displacement amplification (MDA) using the Phi29 DNA polymerase. The generated MDA product was afterwards used for 16S rRNA gene sequence analysis and shotgun-cloned for additional genomic analysis. Sequence analysis showed >99% 16S rRNA gene homology to soil crenarchaeotal clone SCA1170 and shotgun fragments had the closest match to a crenarchaeotal BAC clone previously retrieved from a soil sample. The system was validated using Methanothermobacter thermoautotrophicus as single-cell test organism, and the validation setup produced 100% sequence homology to the ten tested regions of the genome of this organism.