Morphological,Physiological, and Molecular Characterization of a Newly Isolated Steroid-Degrading Actinomycete,Identified as <Emphasis Type="Italic">Rhodococcus ruber</Emphasis> Strain Chol-4

The aerobic degradation of cholesterol, testosterone, androsterone, progesterone, and further steroid compounds as sole carbon source has been observed in the newly isolated bacterial Gram-positive strain Chol-4. The 16S rRNA gene sequence shares the greatest similarity with members of the genus Rhodococcus, with the closest shared nucleotide identities of 98–99% with Rhodococcus ruber (DSM 43338^T) and Rhodococcus aetherivorans (DSM 44752^T). Phylogenetic analysis of Rhodococcus 16S rRNA gene sequences consistently places strain Chol-4 in a clade shared with those both type strains within the Rhodococcus rhodochrous subclade. The results of DNA–DNA hybridization against its two phylogenetically closest neighbors as well as the results of morphological, physiological, and biochemical tests allowed genotypic and phenotypic differentiation of strain Chol-4 from Rhodococcus ruber (DSM 43338^T) on the species level and from the other validly described Rhodococcus species on the genus level. Strain Chol-4 therefore merits recognition as a novel strain of the species Rhodococcus ruber and demonstrates for the first time the capability of this species to utilize a great variety of steroid compounds as growth substrates never shown for other species of this genus so far. The genome of strain Chol-4 harbors at least one gene cluster that may be responsible for the degradation of steroid compounds. This gene cluster was identified in a cloned 5458 bp BamHI–EcoRV DNA fragment and compared to similar genes from other Gram-positive and Gram-negative bacteria described so far.     

Development of species‐specific primers for rapid diagnosis of Tetranychus urticae,T. kanzawai,T. phaselus and T. truncatus (Acari: Tetranychidae)

Species diagnosis is of the utmost importance to both pest management and plant quarantine services. Because of difficulties in the morphological diagnosis of spider mites, molecular techniques are of great value to rapidly and accurately diagnose closely related species. We examined four species of genus Tetranychus (the green and red forms of T. urticae, and T. kanzawai, T. phaselus and T. truncatus), which are found in Korea and are of significance to plant quarantine services. DNA samples isolated from a single egg, larva or adult weighed 64–188 ng. We designed species‐specific primers by performing sequence alignment for 107 sequences of the ribosomal internal transcribed spacer 2 (ITS2) region, which we obtained from GenBank, and sequences generated in this study. Specific nucleotides of each species were selected for designing primers specific for each species. Each species‐specific primer pair, when used to perform PCR analyses, detected only the species from which it originated. However, a T. urticae‐specific primer pair did not discriminate between the green and red forms of this species. These species‐specific primers can be applied in practice for the rapid and accurate diagnosis of spider mite species in plant quarantine and in agricultural fields.     

Three new species,a new combination,and a neotypification in <Emphasis Type="Italic">Dahlstedtia</Emphasis> (Leguminosae,Millettieae, Papilionoideae) from South America

Three new species of Dahlstedtia, D. burkartii, D. dehiscens, and D. lewisiana, are described from South America, and their relationships with related species are discussed. Dahlstedtia burkartii, from Argentina, has pink flowers with standard straight, whereas D. dehiscens and D. lewisiana, both from Brazil, have purplish to lilac flowers with a reflexed standard petal. A new combination, Dahlstedtia peckoltii, is proposed based on Lonchocarpus peckoltii and a neotype is selected for the latter. Information about geographic distribution and phenology of the species is provided.     

Analysis of polyphenols,phytosterols, and bitter principles in <Emphasis Type="Italic">Teucrium</Emphasis> L. species

Total polyphenols, tannins, β-sitosterol, and bitterness values were determined in native and cultivated populations of Teucrium L. species from Croatia (T. arduini L., T. botrys L., T. chamaedrys L., T. flavum L., T. montanum L., T. polium L., and T. scordium L. subsp. scordioides Schreb.). Teucrium species have long been present in folk medicine for diverse medicinal uses, but little is known about polyphenols, phytosterols, and bitter principles. Spectrophotometrically determined contents of total polyphenols (TP) and tannins (T) significantly varied among investigated Teucrium species and were somewhat higher in native populations. The highest TP and T contents were measured in native specimens of T. montanum (TP: 13.68%; T: 3.48%). Scanning densitometry was used for measurements of β-sitosterol levels in plant samples ranged from 0.056% (T. montanum) to 0.129% (T. botrys), and it was not significantly different between native and cultivated specimens of the same plant species. Bitterness values were similar for native and cultivated samples of the same plant species; the highest was measured for T. montanum (15659). The present study suggests that Teucrium species growing in Croatia have potential for cultivation and might be a valuable source of natural bioactive compounds.     

Bees of the genus <Emphasis Type="Italic">Rophites</Emphasis> Spinola (Hymenoptera,Halictidae, Rophitinae) of Russia and adjacent territories

An illustrated key to Rophites species and an annotated list of 9 species from Russia and adjacent territories are given. Rophites clypealis stepposus subsp. n. and the female of R. schoenitzeri Dubitzky, 2004 are described. Lectotypes of Rhophites caucasicus Morawitz, 1876 and Rh. mandibularis Morawitz, 1891 are designated. Rophites algirus is recorded for the first time from Armenia; R. caucasicus, from Azerbaijan; R. clypealis, from Bulgaria, Russia, and Kazakhstan; R. foveolatus, from Georgia; R. quinquespinosus, from Armenia and Uzbekistan; R. schoenitzeri, from Azerbaijan and Iran.     

Cryptic diversity,biogeography and genetic variation in Northeast Pacific species of <Emphasis Type="Italic">Porphyra</Emphasis> sensu lato (Bangiales,Rhodophyta)

We sequenced the chloroplast rubisco large subunit (rbcL) gene in 236 samples of Porphyra sensu lato from the northeast Pacific. Comparisons of sequences within the study area as well as comparisons with published sequences revealed up to five cryptic species among the 22 named species: a species closely related to Porphyra abbottiae, a species previously identified as P. pseudolinearis, a species closely related to P. pseudolanceolata and previously identified as that species, a previously unknown species from the eastern Aleutian Islands, and a species closely related to P. schizophylla and previously identified as that species. All of these previously unrecognized species had high bootstrap values separating them from the other species. In addition, our wide geographic sampling allowed us to extend, curtail or clarify the geographic ranges of a number of the species. We also provide published sequences for P. gardneri and P. smithii for the first time. We compared amount of sequence divergence within species grouped on the basis of sexuality (monoecious, sectored into separate male and female “halves”, or dioecious), habitat (high, mid, or low intertidal/subtidal), and seasonality (winter, spring, or summer) using Tukey’s HSD t test, but we observed no significant differences between species grouped in this manner. Different species showed different levels of genetic variation in the rbcL gene apparently unrelated to these traits. Also, we observed no differences in the patterns of genetic variation in a species based on whether the specimens were collected from outside or from within the region covered by ice during Pleistocene glaciations.     

Phylogeny of Klebsormidium (Kelbsormidiales; Charophyta) Based on rbcL,atpB, coxIII and nad5 Sequence Data

The charophycean order Klebsormidiales consists of two unbranched filamentous lineages, the fresh water Entransia and the semiterrestrial Klebsormidium. Molecular data have supported the close relationship Klebsormidiales have with land plants. A monograph exists for the European species of Klebsormidium; this monograph provides a cladistic analysis of morphological characteristics for eight Klebsormidium species, but a species level molecular phylogenetic analysis has not yet been performed for the group. We have obtained 50 strains of Klebsormidium, representing 11 morphological species, both from nature and from culture collections. Phylogenetic analyses using rbcL, atpB, coxIII and nad5 gene sequence data from these strains and 40 outgroup sequences suggest that several traditional Klebsormidium species may not be monophyletic.     

A review of the tribe Phanerotomini (Hymenoptera,Braconidae, Cheloninae) in Turkey,with a new host record for <Emphasis Type="Italic">Phanerotoma</Emphasis> (<Emphasis Type="Italic">Bracotritoma</Emphasis>) <Emphasis Type="Italic">permixtellae</Emphasis>

In this study, the Phanerotomini of Turkey collected between July 1959 and July 2004, are listed. A total of thirteen Phanerotomini species were determined, and six of these are reported for the first time from Turkey. Phanerotoma (Bracotritoma) permixtellae Fischer, 1968 was reared from a new natural host Recurvaria pistaciicola (Danilevskii, 1955) (Lepidoptera: Gelechiidae). With this present study an illustrated identification key for the Phanerotomini of Turkey is given. The distribution of the Phanerotomini species is discussed according to the phytogeographical provinces of Turkey.     

Ophiostomatoid fungi associated with the northern spruce engraver, <Emphasis Type="Italic">Ips perturbatus</Emphasis>, in western Canada

A number of ophiostomatoid fungi were isolated from the spruce-infesting bark beetle, Ips perturbatus and its galleries collected from felled spruce trees and logs in northern BC and the Yukon Territory. Isolates were identified to species using morphological characteristics, nuclear ribosomal DNA and partial β-tubulin gene sequences. Thirteen morphological and phylogenetic species were identified among the isolates. Leptographium fruticetum, Leptographium abietinum, Ophiostoma bicolor, Ophiostoma manitobense, O. piceaperdum, and eight undescribed species of the genus Ophiostoma and the anamorph genera Leptographium, Hyalorhinocladiella, Ambrosiella and Graphium. A number of these species, i.e. L. fruticetum, Hyalorhinocladiella sp. 2, O. bicolor and O. manitobense, were isolated repeatedly from I. perturbatus, while others, i.e. Graphium sp. 1 and O. piceaperdum, seemed to be␣sporadic associates. Among all the isolates, L. fruticetum had the highest relative dominance in this survey. A high frequency of occurrence of this species with the beetle may indicate a specific relationship between the two partners.     

Contribution to the taxonomy,distribution and paleoecology of the early representatives of <Emphasis Type="Italic">Penthesilenula</Emphasis> Rossetti &; Martens, 1998 (Crustacea,Ostracoda, Darwinulidae) from Argentina,with the description of a new species

Early Mesozoic Argentinian darwinuloids, now re-assigned to the genus Penthesilenula (incae group), are described and illustrated. These records seem to constitute the oldest known for the genus. The close similarities between the studied species and those described from eastern Asia enlarge the geographical distribution of Penthesilenula (incae group), which, although restricted to the southern Hemisphere in Recent and Holocene times, may have had a wider distribution in the past. Additional information on the paleoecology, plus a preliminary account on the distribution of darwinulid species from Argentina are also offered. One new species is described: Penthesilenula loana sp. nov.     

Two new species of <Emphasis Type="Italic">Cotingacola</Emphasis> Carriker,Phthiraptera: Ischnocera: Philopteridae) from Amazonian Brazil,with comments on host-specificity

This paper describes two new species of Cotingacola Carriker, 1956, C. lutzae n. sp. and C. engeli n. sp., from the cinereous mourner Laniocera hypopyrra (Vieillot) and black-necked red cotinga Phoenicircus nigricollis Swainson (Aves: Passeriformes: Cotingidae), respectively. These species are fully illustrated and compared with their morphologically closest relatives. With the addition of these two new species, this Neotropical chewing louse genus now includes ten species. We also present records from Brazil for two previously described species, C. stotzi Clayton & Price, 1998 and C. parmipapillae Carriker, 1956. The re-examination of a specimen identified as ‘Cotingacola sp.’ in a recent phylogenetic study confirmed that it is C. parmipapillae and that Querula purpurata (Statius Müller) regularly hosts two Cotingacola species, which means that at least three cotingid hosts carry more than one species of Cotingacola. We comment on the variability in the host-specificity of Cotingacola species.     

Restoring <Emphasis Type="Italic">Daphnia lacustris</Emphasis> G.O. Sars, 1862 (Crustacea,Anomopoda): a cryptic species in the <Emphasis Type="Italic">Daphnia longispina</Emphasis> group

While molecular markers have revealed several distinct species within the Daphnia longispina group, there is a need to reconcile these species with traditional nomenclature. Here we show that one such species, called D. longispina in recent literature based on molecular markers, can reliably be associated with the described taxon Daphnia lacustris G.O. Sars, 1862. Both mitochondrial and nuclear molecular markers readily distinguish this species from others in the D. longispina group. D. lacustris is absent in the region from which D. longispina was first described (Denmark), and the designation D. longispina must be reserved for another widespread species represented by Danish lineages. While the diagnosis of D. lacustris (and other species of the D. longispina group) by molecular markers is unequivocal, distinguishing it morphologically from other species is still problematic. The presently known distribution range of D. lacustris includes most of Norway, northern Finland and a single lake in the Polish Tatra Mountains. Its typical habitat is oligotrophic lakes without intense fish predation. Guest editor: Piet Spaak Cladocera: Proceedings of the 7th International Symposium on Cladocera     

Mating Behavior and Species Status of Host-Associated Populations of the Polyphagous Thrips, <Emphasis Type="Italic">Frankliniella schultzei</Emphasis>

The thrips taxon, Frankliniella schultzei Trybom, could possibly comprise a complex of unrecognised cryptic species. We therefore made a range of reproductive behavioral observations on F. schultzei individuals from different host-associated populations to investigate their species status. We first described the mating behavior of F. schultzei taken from flowers of a primary host species, Malvaviscus arboreus. The pattern of male-female interaction was complex and protracted, with several behavioral steps being performed. These steps were similar for all mating pairs. Most females responded to male presence before physical contact, suggesting that male pheromones may be involved. We then compared mating behaviors, brood sizes and sex ratios of F. schultzei pairs derived from different host plant species. All pairs in crosses regardless of their original host plant species produced female offspring, indicating successful mating. Frequencies of behaviors were similar for all pairs. Only those crosses involving F. schultzei individuals from Erythrina crista-galli flowers differed from the general pattern in pre-copulation and copulation times. The females from E. crista-galli also produced significantly fewer offspring than females from other crosses. Finally we used traps baited with males from two populations, but set among flowers of one host plant species, to test for differential attraction of females. We found no evidence that the different host-associated populations investigated differ from one another functionally in their sex pheromone system. However, these results do not yet discount the possibility of cryptic species within the taxon F. schultzei. Populations of F. schultzei on many other host species need to be investigated from this perspective.     

Sequence and RFLP analysis of the ITS2 ribosomal DNA in two Neotropical social bees, <Emphasis>Melipona beecheii</Emphasis> and <Emphasis>Melipona yucatanica</Emphasis> (Apidae,Meliponini)

Two stingless bees species of the genus Melipona, M. beecheii and M. yucatanica, are the only ones reported for the Yucatan Peninsula. The natural distribution of M. beecheii ranges from southern Mexico to Costa Rica, that of M. yucatanica from south Mexico to Guatemala. Colonies of both species occur in a variety of habitats and show adaptations to local conditions denoting the occurrence of ecotypes. The ITS2 of ribosomal DNA has been characterized in both species and its utility to discriminate among colonies has been investigated through RFLP experiments. The ITS2 region is unusually long, 1788 bp in M. beecheii and 1845 bp in M. yucatanica (including the 3′ end of the 5.8S gene and partial 5′ of the 28S gene). Mean nucleotide divergence between both ITS2 sequences is 16% (excluding sites with insertions/deletions) and 20% when the insertions/deletions are taken into account. The G+C content in both sequences is close to 53%. The PCR-RFLP assay was performed with 12 restriction enzymes on colonies of M. beecheii from Mexico (Yucatan, Campeche and Chiapas) Costa Rica, El Salvador and Guatemala, and of M. yucatanica from Mexico (Yucatan) and Guatemala. The restriction patterns obtained allow to discriminating colonies of both species with different origins. Both kinds of data are thus useful for assessing intra and interspecific genetic variability and for developing appropriate conservation strategies for these species. Received 20 June 2007; revised 31 August 2007; accepted 12 September 2007.     

Two new species of <Emphasis Type="Italic">Anthobothrium</Emphasis> van Beneden, 1850 (Tetraphyllidea: Phyllobothriidae) from carcharhinid sharks,with a redescription of <Emphasis Type="Italic">Anthobothrium laciniatum</Emphasis> Linton, 1890

Anthobothrium laciniatum Linton, 1890 is redescribed based on specimens taken from the dusky shark Carcharhinus obscurus (Lesueur) collected from the Northwestern Atlantic Ocean, and a neotype is designated. A. laciniatum differs from A. cornucopia van Beneden, 1850, A. altavelae Euzet & Ben Hassine, 2002, A. lesteri Williams, Burt & Caira, 2004 and A. spinosum Subhapradha, 1955 in total length. It further differs from A. cornucopia, A. altavelae and A. spinosum in proglottid number, and differs from A. galeorhini Suriano, 2002, A. cornucopia, and A. spinosum in testis number. A. lyndoni n. sp. is described from the sandbar shark C. plumbeus (Nardo). This new species differs from A. laciniatum in ovarian width and from A. cornucopia, A. altavelae, A. galeorhini and A. spinosum in the total number of proglottids. It further differs from A. cornucopia, A. galeorhini, and A. spinosum in total length, and from A. cornucopia and A. galeorhini in the number of testes. A. lyndoni n. sp. differs from A. lesteri in bothridial muscle and ovarian morphology. Anthobothrium caseyi n. sp. is described from Prionace glauca (Linnaeus). This new species differs conspicuously from the other six species of Anthobothrium van Beneden, 1850 (sensu stricto) in the shape of its proglottid laciniations. The taxonomic status of 43 species that have been associated with Anthobothrium is addressed. Taxonomic actions regarding Anthobothrium during the past century have resulted in a polyphyletic taxon.     

The discovery of <Emphasis Type="Italic">Caligus</Emphasis><Emphasis Type="Italic">temnodontis</Emphasis> Brian, 1924 (Copepoda: Caligidae) from the bluefish <Emphasis Type="Italic">Pomatomus saltatrix</Emphasis> (Linnaeus) in the eastern Mediterranean Sea

The temnodontis variety of Caligus mauritanicus Brian, 1924 described by Brian (1924) is a valid species known only from a single host species, Pomatomus saltatrix (Linnaeus). New material of this species has been examined from the same host fish caught from Abuqir Bay, Alexandria (Egypt), from Iskenderun Bay (Turkey) and from off the coast of South Africa. Using this material, C. temnodontis Brian, 1924 is redescribed and compared with related species. It is most closely related to the Indo-Pacific species C. pagrosomi Yamaguti, 1939.