A diazotrophic bacterial endophyte isolated from stems of Zea mays L. and Zea luxurians Iltis and Doebley

The apoplastic fluids of field-grown Zea mays and Zea luxurians plants were isolated from surface sterilized stem tissue by centrifugation and spread on agar plates containing a nitrogen-free, defined medium. The predominant bacterium isolated from these plates was characterized further. The ability of this bacterium to fix nitrogen was confirmed by its ability to grow on a semi-solid, nitrogen-free medium and reduce ¹⁵N₂ to ¹⁵NH₃ and acetylene to ethylene. Protions of the nifH and 16S rRNA genes from this organism were amplified by PCR and sequenced. The nifH gene, which codes for dinitrogenase reductase, from this organism is closely related to nifH from Klebsiella pneumoniae. Similarly, the 16S rRNA gene sequences and carbon utilization tests grouped it closely with K. pneumoniae. Based an these data, the isolates from Z. mays and Z. luxurians are tentatively classified as Klebsiella spp. (Zea). The ability of this bacterium to contribute to the nitrogen economy of the corn plant is unknown.     

PEDINOMONAS NOCTILUCAE (PRASINOPHYCEAE), THE FLAGELLATE SYMBIOTIC IN NOCTILUCA (DINOPHYCEAE) IN SOUTHEAST ASIA1

The small green flagellate symbiotic in the Noctiluca miliaris Suriray from Southeast Asia has been examined by light and electron microscopy. The flagellate is very similar to Pedinomonas minor Korschikoff. The deeper flagellar depression and the habitat distinguish this species of Pedinomonas from P. minor. It is not a euglenoid as originally proposed, since it contains starch. Characters distinguishing it from Micromonas are described. The new combination Pedinomonas noctilucae (Subrahmanyan) comb, nov. is proposed for this flagellate.     

Trichodectes canis (De Geer, 1778) (Phthiraptera, Ischnocera), a new ectoparasite of the raccoon dog (Nyctereutes procyonoides) in the Czech Republic

The raccoon (Nyctereutes procyonoides) is described as a host of the dog biting louse, Trichodectes canis (De Geer, 1778). The present note reports the occurrence of this species of louse on two raccoon dogs from Moravia and Bohemia. The specimens of T. canis collected on this host differed from T. canis originating from the domestic dog by body size only.     

Mode of life of the Silurian uncoiled gastropod Semitubina sakoi n. sp. from Japan

Semitubina sakoi n. sp. from the late Silurian of Japan represents the second species of this genus and also the first record of a Silurian gastropod in Japan. The gastropod shells occur in a thin mudstone bed and were found to be encrusted exclusively by corallites of ? Favosites sp. These corallites reveal that encrustation proceeded as the gastropod shells grew. The ecological relationship between the two organisms is considered to be symbiotic. This mode of life allowed the coral to live on a muddy substrate because clear sea water passed over the colony as the gastropod moved along. The gastropod benefited from this relationship by being protected from shell-boring or shell-crushing predators by the encrusting corallite. In Semitubina sakoi the body whorl is separated from the penultimate one by a considerable gap in a later growth stage and S. sakoi has been cited as one of the uncoiled gastropods. The uncoiling of this gastropod results primarily from rapidly increasing whorl translation rate in the latest growth stage. Taking the symbiotic relationship with ? Favosites sp. into consideration, a deposit feeding or benthic scavenging mode of life is suggested for this gastropod.     

Gram positivity induced in Escherichia coli by chloroform-methanol treatment

Summary Extraction of lipid by chloroform-methanol from Escherichia coli induces Gram positivity in the cells of this bacterium. It has been observed that approximately 1.5 times more lipid is extracted from Escherichia coli than from Staphylococcus aureus by this treatment. An increase of 1.5–1.8 fold in the retention of dye has been evident in the case of S. aureus where as this was 70–90 fold in the case of E. coli, which is 1/4th of the normal retention by S. aureus. The more lipid present in the cell wall or cell membrane, the less retention of dye seems to be exhibited by the bacteria. This also substantiates our model for Gram reaction presented in a previous communication.     

Nest-moving by the polydomous ant <Emphasis Type="Italic">Cataglyphis iberica</Emphasis>

In this paper we analyze emigration from nests by the polydomous ant Cataglyphis iberica. Social carrying of workers of this species between different nests of the colony is frequent. In Bellaterra (Barcelona, NE Spain), we monitored field emigration of C. iberica by noting for each nest the migratory behavior of C. iberica workers and, when the nests were attacked by another ant species, Camponotus foreli, we noted the number of C. foreli workers involved in the attacks. Emigration of C. iberica from nests was highly variable. We suggest the main factor determining emigration by this species was attack by workers of C. foreli, so emigration from C. iberica nests was much faster when harassment by C. foreli increased. The system of multiple nests of C. iberica enables this species to abandon attacked nests and to reinstall their population in other nests of the same colony. This reduces risk to the colonies because the route between the different nests is well known by transporter workers.     

New finds of the Neogene lizard and snake fauna (Squamata: Lacertilia; Serpentes) from the Slovak Republic

This paper deals with the squamate fauna from three Neogene localities in Slovakia. Neogene lizards and snakes have rarely been reported from this region and many aspects of their evolution and palaeodiversity are still poorly understood. Squamate remains from the Upper Miocene (MN 9) locality of Borsky Sv?ty Jur belong to at least five different taxa: Lacerta sp., Pseudopus sp., Colubroidea indet. (? Elapidae), Natricinae indet. and Colubridae indet. The ophidian assemblage from Ivanovce (Lower Pliocene; MN 15b) is dominated by colubrids, mainly Zamenis longissimus. The remainder is comprised of the species Natrix natrix. Squamate material from this locality also includes Lacerta cf. agilis, Pseudopus sp. and Ophisaurus sp., while that from Hajnáčka (Upper Pliocene; MN 16a) is extremely poor, comprising just one taxon - Natrix natrix. The Ivanovce material represents the oldest proof of the former existence of the species Zamenis longissimus and Natrix natrix in the Slovak Republic. The composition of this snake fauna indicates faunistic and palaeoecological changes at the end of the Miocene, although this was not as rapid as at similar localities in Europe. The squamate fauna of Slovakia has changed very little since the beginning of the Pliocene, and the majority of taxa are represented by species that currently occur in this region.     

Milk Clotting Enzyme from Microorganisms

The purification of the milk clotting enzyme from Mucor pusillus Lindt could be achieved by column chromatography on Amberlite IRC-50 by raising pH from 3.5 to 4.5 and about 70% of activity was recovered after this treatment. After the treatment through the column of DEAE-Sephadex A-25, the trace cellulase activity could be eliminated.

The homogeneity of the purified preparation was proved by ultracentrifugal analysis and electrophoretic patterns at various pH values.

Isoelectric point of this enzyme is considered to lie between pH 3.5 and 3.8.

The enzyme activity was inhibited by Hg⁺⁺ or Fe⁺⁺⁺.

Trypsinogenkinase activity was not contained in this enzyme.

The antiserum against the milk clotting enzyme from Mucor pusillus reacted with the purified and crude enzyme preparations in precipitin test and inhibited their enzyme activities, but did not react with other enzymes such as rennin, pepsin, acid proteases from Aspergillus saitoi and Aspergillus oryzae, or the culture filtrates of some strains of Mucor and Rhizopus.

The antigen-antibody reaction was so specific that it might be possible with this antibody to identify this enzyme and also the strain itself.

Normal sera from some mammals inhibited this enzyme activity too, but the degree was less than that with rennin.     

Salsuginus neotropicalis n. sp. (Monogenea: Ancyrocephalinae) from the pike killifish Belonesox belizanus (Atheriniformes: Poeciliidae) from southeastern Mexico

Salsuginus neotropicalis n. sp. is described from the gills of Belonesox belizanus (Kner), the pike killifish (Poeciliidae), from the southeastern Mexico. S. neotropicalis is distinguished from all other congeners by the morphology of the haptoral sclerites, especially by the presence of a ventral bar with notably enlarged ends, and by possessing overlapping gonads. The diagnosis of Salsuginus is amended based on the position of gonads in this species. This is the first known Salsuginus species occurring in a freshwater fish from the Neotropical Region.     

The effect of heated effluent on the occurrence and the reproduction of the freshwater limpetsAncylus fluviatilis Müller, 1774,Ferrissia wautieri (Mirolli, 1960) andAcroloxus lacustris (L., 1758) in two Dutch water bodies

The discharge of cooling water favours the occurrence of freshwater limpets. Reproduction ofAncylus fluviatilis is stimulated by the higher temperature of the cooling water; the life cycle ofFerrissia wautieri (nearly exclusively found in the outlet areas) does not, however, differ from that under natural temperature regimes. ForAcroloxus lacustris this is not clear as detailed studies on the life cycle under natural temperature circumstances are lacking. Acroloxus lacustris andFerrissia wautieri could be found in large numbers on hard substrates (stones), as well as on plant material in sheltered as well as in exposed situations. In this respect they differ fromAncylus fluviatilis, which is restricted in its occurrence to hard substrates and exposed situations.It is not very clear whether the occurrence ofAncylus fluviatilis is also influenced by some form of competition for food byBithynia tentaculata; both feed on diatoms. Further experimental studies of this question should yield interesting results.The changes in numbers of the three freshwater limpet species are mainly due to reproduction and migration, both largely governed by temperature and day length.It was possible to reconstruct the influence of cooling water in the Bergumermeer area by means of the ratio between the numbers ofAcroloxus lacustris andFerrissia wautieri. The share ofFerrissia was highest near the outiet and decreased with the distance from this station.     

New Species of Spirotrichonympha from Reticulitermes and the Relationships Among Genera in Spirotrichonymphea (Parabasalia)

Spirotrichonymphea is a class of hypermastigote parabasalids defined by their spiral rows of many flagella. They are obligate hindgut symbionts of lower termites. Despite more than 100 yr of morphological and ultrastructural study, the group remains poorly characterised by molecular data and the phylogenetic positions and taxonomic validity of most genera remain in question. The genus Spirotrichonympha has been reported to inhabit several termite genera, including Reticulitermes, Coptotermes, and Hodotermopsis. The type species for this genus, Spirotrichonympha flagellata, was described from Reticulitermes lucifugus but no molecular data are yet available for this species. In this study, three new Spirotrichonympha species are described from three species of Reticulitermes. Their molecular phylogenetic position indicates that the genus is not monophyletic, as Spirotrichonympha species from Coptotermes, Paraneotermes, and Hodotermopsis branch separately. In contrast, the genus Holomastigotoides is monophyletic, as demonstrated using new sequences from Holomastigotoides species. The presence of Holomastigotoides in Prorhinotermes and the distinct phylogenetic positions of Spirotrichonympha from Reticulitermes and Coptotermes are consistent with a previously proposed symbiont fauna replacement in the ancestor of Reticulitermes.     

Identification and characterization of a bacteria-like sequence in the genome of some Silene species

The aim of this work was to characterize a nucleotide sequence MK14 that originated from a plasmid library obtained via degenerate oligonucleotide primed polymerase chain reaction (DOP-PCR) amplification of laser microdissected Y-chromosomes of Silene latifolia. This sequence showed significant similarity to parts of two adjoining genes from bacterial representatives of the genus Ralstonia. MK14 sequence contains a part of a conserved domain, and phylogenetic analysis based on this region confirmed its relationship to Ralstonia-derived sequences. Genomic Southern blot analysis proved the presence of this fragment in the genome of S. latifolia. We hypothesize that this insertion is of bacterial origin, and was probably gained via horizontal gene transfer. Moreover, MK14 insertion is shared by some closely related Silene species, suggesting an ancient spontaneous transformation by an ancestor of bacteria from the genus Ralstonia.     

Redescription of <Emphasis Type="Italic">Amioides grossidens</Emphasis> Smith and Radcliffe 1912 as a valid genus and species of the family Acropomatidae (Perciformes)

The acropomatid, Amia (Amioides) grossidens Smith and Radcliffe 1922, was originally described from a single specimen collected off Luzon Island, the Philippines. Although this species was regarded as belonging to Synagrops, it is distinguished from Synagrops by the dentition of the upper jaw, number of dorsal fin spines, and additionally from other acropomatid genera by 14 caudal vertebrae and a posttemporal spine. Therefore, this species should be placed in a monotypic genus Amioides, with elevation of the subgenus to generic rank, compared with diagnostic characters of seven other genera, and this genus and species were herein redescribed based on the holotype.     

Cloning and characterization of an ftsZ homologue from a bacterial symbiont of Drosophila melanogaster

A 1194 by open reading frame that codes for a 398 amino acid peptide was cloned from a gt11 library of Drosophila melanogaster genomic DNA. The predicted peptide sequence is very similar to three previously characterized protein sequences that are encoded by the ftsZ genes in Escherichia coli, Bacillus subtilis and Rhizobium meliloti. The FtsZ protein has a major role in the initiation of cell division in prokaryotic cells. Using a tetracycline treatment that eradicates bacterial parasites from insects, the ftsZ homologue has been found to be derived from a bacterium that lives within the strain. However, polymerase chain reaction (PCR) amplification of the gene from treated embryos suggests that it is not derived from a gut bacterium. Nevertheless, by amplifying and characterizing part of the 16S rRNA from this bacterium we have been able to demonstrate that it is a member of the genus Wolbachia, a parasitic organism that infects, and disturbs the sexual cycle of various strains of Drosophila simulans. We suggest that this ftsZ homologue is implicated in the cell division of Wolbachia, an organism that fails to grow outside the host organism. Sequence and alignment analysis of this ftsZ homologue show the presence of a potential GTP-binding motif indicating that it may function as a GTPase. The consequences of this function particularly with respect to its role in cell division are discussed.     

Transposon-like structure of a new plasmid-encoded restriction-modification system in Rhizobium leguminosarum VF39SM

Total DNA isolated from Rhizobium leguminosarum VF39SM cells is resistant to cleavage by the restriction endonuclease PstI. Plasmid curing and transfer studies localized this phenotype to pRleVF39b, the second smallest of six plasmids found in this bacterium. In vitro selection for vector modification was employed to isolate a presumptive methylase gene (M.Rle39BI) from a plasmid gene library. Total and plasmid DNAs isolated from E. coli containing M.RleBI were resistant to digestion by PstI. Sequence data suggested that a putative restriction endonuclease (R.Rle39BI) was also encoded on the same fragment. The two genes were flanked by identical copies of a putative insertion sequence, which was also present in several copies elsewhere in the VF39SM genome. The presence of this element in other strains examined suggested that this element is indeed an insertion sequence. The differences in G/C content between the DNA coding for the R/M system and that of the IS element suggest that this DNA region may have been acquired by horizontal transfer. Received: 28 January 1997 / Accepted: 3 June 1997     

Disjunctive distribution of the basal genus Aplomerus (Hymenoptera: Ichneumonidae: Xoridinae) in East Asia and North America,with a new species from Japan

The first Japanese representative of the predominantly Nearctic genus Aplomerus Provancher, A. japonicus sp. nov., is described. This species can easily be distinguished from the other species by the black body. Morphologically this species resembles A. lineatulus (Say) from eastern North America but is separable by the sculpture of the propodeum and third metasomal tergite. The discovery of this species demonstrates a disjunctive distribution of the genus Aplomerus in East Asia and North America. Notes on habitat and adult behavior are also given.     

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.