Divergence within the marker region of the <Emphasis Type="Italic">groESL</Emphasis> operon in <Emphasis Type="Italic">Anaplasma phagocytophilum</Emphasis>

Anaplasma phagocytophilum is an obligate intracellular bacterial parasite in human and animal granulocytes. In Europe, A. phagocytophilum is transmitted by Ixodes ticks; Ixodes ricinus is the vector of the parasite in Poland. In terms of epidemiology, the identification of pathogens in ticks increasingly relies on molecular techniques. Polymerase chain reaction (PCR) with species-specific primers is a tool that allows the quick and accurate detection of pathogens in ticks, humans, or animals. DNA was extracted from the blood of Capreolus capreolus and Cervus elaphus, and amplified using the primers HS1/HS6 (external) and HS43/HSVR (internal). For sequencing, six samples from roe deer and two samples from red deer were selected, and the resulting sequences were submitted to GenBank (accession numbers DQ779568, DQ779567, EU157919, EU157920, EU157921, EU157922). These nucleotide sequences were compared with each other and five variants were distinguished in roe deer and one in red deer. A comparison of the sequences of the author’s database revealed 45 polymorphic sites of substitution character (76% transitions and 24% transversions). The homology tree revealed two groups, one with sequences only from roe deer, while the second with sequences isolated mainly from red deer, livestock animals, and humans. These strains of A. phagocytophilum are also present in Poland.     

Chromosome pairing in allotetraploid hybrids of <Emphasis Type="Italic">Festuca pratensis</Emphasis> × <Emphasis Type="Italic">Lolium perenne</Emphasis> revealed by genomic <Emphasis Type="Italic">in situ</Emphasis> hybridization (GISH)

Genomic in situ hybridization (GISH) was used to make a detailed study of chromosome pairing at metaphase I (MI) of meiosis in six F(1) hybrid plants of the allotetraploid Festuca pratensis x Lolium perenne (2n = 4x = 28; genomic constitution FpFpLpLp). The mean chromosome configurations for all hybrids analysed were 1.13 univalents + 11.51 bivalents + 0.32 trivalents + 0.72 quadrivalents, and the mean chiasma frequency was 21.96 per cell. GISH showed that pairing was predominantly intragenomic, with mean numbers of L. perenne (Lp/Lp) and F. pratensis (Fp/Fp) bivalents being virtually equal at 5.41 and 5.48 per cell, respectively. Intergenomic pairing between Lolium and Festuca chromosomes was observed in 33.3% of Lp/Fp bivalents (0.62 per cell), in 79.7% of trivalents - Lp/Lp/Fp and Lp/Fp/Fp (0.25 per cell), and in 98.4% of quadrivalents - Lp/Lp/Fp/Fp and Lp/Lp/Lp/Fp (0.71 per cell). About 4.0% of the total chromosome complement analysed remained as univalents, an average 0.68 Lp and 0.45 Fp univalents per cell. It is evident that in these hybrids there is opportunity for recombination to take place between the two component genomes, albeit at a low level, and this is discussed in the context of compromising the stability of Festulolium hybrid cultivars and accounting for the drift in the balance of the genomes over generations. We speculate that genotypic differences between hybrids could permit selection for pairing control, and that preferences for homologous versus homoeologous centromeres in their spindle attachments and movement to the poles at anaphase I could form the basis of a mechanism underlying genome drift.     

Simultaneous real-time PCR detection of <Emphasis Type="Italic">Bacillus anthracis</Emphasis>, <Emphasis Type="Italic">Francisella tularensis</Emphasis> and <Emphasis Type="Italic">Yersinia pestis</Emphasis>

This report describes the development of in-house real-time PCR assays using minor groove binding probes for simultaneous detection of the Bacillus anthracis pag and cap genes, the Francisella tularensis 23 KDa gene, as well as the Yersinia pestis pla gene. The sensitivities of these assays were at least 1 fg, except for the assay targeting the Bacillus anthracis cap gene, which showed a sensitivity of 10 fg when total DNA was used as a template in a serial dilution. The clinical value of the Bacillus anthracis- and Francisella tularensis-specific assays was demonstrated by successful amplification of DNA from cases of cow anthrax and hare tularemia, respectively. No cross-reactivity between these species-specific assays or with 39 other bacterial species was noted. These assays may provide a rapid tool for the simultaneous detection and identification of the three category A bacterial species listed as biological threats by the Centers for Disease Control and Prevention.     

Collar spine models in the genus <Emphasis Type="Italic">Echinostoma</Emphasis> (Trematoda: Echinostomatidae)

This paper discusses collar spine arrangements in the genus Echinostoma. All arrangements are of uneven numbers of collar spines on the oral collar. The total number of collar spines in these arrangements ranges from a low 31 to a high 51. There are 11 models of collar spine arrangements in the Echinostoma consisting of spine numbers 31, 33, 35, 37, 39, 41, 43, 45, 47, 49, and 51. Representative species with these collar spine arrangements are given in the article. The number of collar spines in a species is identical in both the larval and adult forms. Reports of even numbered spine counts in the genus Echinostoma are erroneous and probably reflect counts on worms with lost, retracted, or supernumerary spines.     

Redescription of <Emphasis Type="Italic">Decorataria decorata</Emphasis> (Spirurida,Acuariidae) based on nematodes from <Emphasis Type="Italic">Podiceps cristatus</Emphasis> and <Emphasis Type="Italic">P. grisegena</Emphasis> (Aves,Podicipediformes) from Bulgaria

Decorataria decorata (Cram, 1927) is redescribed on the basis of light-microscopy and SEM observations on specimens collected from the stomach of Podiceps cristatus and P. grisegena from Bulgaria. The SEM study revealed the presence of a porebearing field on each pseudolabium and a pair of spines (one dorsal and one ventral) situated between bases of the cordons. The deirids are spine-like and minute. The light-microscopy examination showed the presence of ornamentation situated under the dorsal surface of caudal alae. The occurrence of D. decorata in Bulgaria is a new geographical record.     

SCC<Emphasis Type="Italic">mec</Emphasis> and <Emphasis Type="Italic">spa</Emphasis> types of methicillin-resistant <Emphasis Type="Italic">Staphylococcus aureus</Emphasis> strains in Israel. Detection of SCC<Emphasis Type="Italic">mec</Emphasis> type V

Molecular characterization of methicillin-resistant Staphylococcus aureus isolates from three hospitals in Israel was the aim of the study presented here. We identified 11 distinct genetic clones by pulsed-field gel electrophoresis. Molecular typing identified four different SCCmec types—I, II, IV, and V−and nine spa types. Spa type t002 was the most common. An erratum to this article can be found at     

<Emphasis Type="Italic">Trichomonas vaginalis</Emphasis> harboring <Emphasis Type="Italic">Mycoplasma hominis</Emphasis> increases cytopathogenicity in vitro

The parasite Trichomonas vaginalis causes one of the most common non-viral sexually transmitted infections in humans. Mycoplasmas are frequently found with trichomonads but the consequences of this association are not yet known. In the present study, the effects of T. vaginalis harboring M. hominis on human vaginal epithelial cells and on MDCK cells are described. The results were analyzed by light, scanning and transmission electron microscopy, as well as using cell viability assays. There was an increase in the cytopathic effects on the epithelial cells infected with T. vaginalis associated with M. hominis compared to T. vaginalis alone. The epithelial cells exhibited an increase in the intercellular spaces, a lesser viability, and increased destruction provoked by the infected T. vaginalis. In addition, the trichomonads presented a higher amoeboid transformation rate and an intense phagocytic activity, characteristics of higher virulence behavior.     

A frameshift mutation of the chloroplast <Emphasis Type="Italic">mat</Emphasis>K coding region is associated with chlorophyll deficiency in the <Emphasis Type="Italic">Cryptomeria japonica</Emphasis> virescent mutant <Emphasis Type="Italic">Wogon</Emphasis>-Sugi

Wogon-Sugi has been reported as a cytoplasmically inherited virescent mutant selected from a horticultural variety of Cryptomeria japonica. Although previous studies of plastid structure and inheritance indicated that at least some mutations are encoded by the chloroplast genome, the causative gene responsible for the primary chlorophyll deficiency in Wogon-Sugi, has not been identified. In this study, we identified this gene by genomic sequencing of chloroplast DNA and genetic analysis. Chloroplast DNA sequencing of 16 wild-type and 16 Wogon-Sugi plants showed a 19-bp insertional sequence in the matK coding region in the Wogon-Sugi. This insertion disrupted the matK reading frame. Although an indel mutation in the ycf1 and ycf2 coding region was detected in Wogon-Sugi, sequence variations similar to that of Wogon-Sugi were also detected in several wild-type lines, and they maintained the reading frame. Genetic analysis of the 19 bp insertional mutation in the matK coding region showed that it was found only in the chlorophyll-deficient sector of 125 full-sibling seedlings. Therefore, the 19-bp insertion in the matK coding region is the most likely candidate at present for a mutation underlying the Wogon-Sugi phenotype. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Catalog of 86 single-nucleotide polymorphisms (SNPs) in three uridine diphosphate glycosyltransferase genes: <Emphasis Type="Italic">UGT2A1</Emphasis>, <Emphasis Type="Italic">UGT2B15</Emphasis>, and <Emphasis Type="Italic">UGT8</Emphasis>

We report here three high-density maps of variations found among 48 Japanese individuals in three uridine diphosphate glycosyltransferase (UGT) genes, UGT2A1, UGT2B15, and UGT8. A total of 86 single-nucleotide polymorphisms (SNPs) were identified through systematic screening of genomic regions containing these genes: 8 in 5′ flanking regions, 7 in coding regions, 67 in introns, 3 in 3′ untranslated regions, and 1 in a 3′ flanking region. We also discovered 14 variations of other types. Of the 86 SNPs, 63 (73%) were considered to be novel on the basis of comparison of our data with the Database of SNPs (dbSNP) of the National Center for Biotechnology Information. Among the seven SNPs identified in exonic sequences, five were non-synonymous changes that would result in amino-acid substitutions. The collection of SNPs derived from this study will serve as an additional resource for studies of complex genetic diseases and responsiveness to drug therapy. Received: June 12, 2002 / Accepted: June 13, 2002     

Genetic diversity of porcine <Emphasis Type="Italic">Norovirus</Emphasis> and <Emphasis Type="Italic">Sapovirus</Emphasis>: Canada, 2005–2007

Noroviruses and sapoviruses are members of the family Caliciviridae and emerging enteric pathogens of humans and animals. Since their discovery and characterization in swine, relatively few strains have been described in detail. In order to investigate their genetic diversity, a total of 266 fecal samples collected in the province of Quebec, Canada, between 2005 and 2007 were screened for the presence of caliciviruses by RT-PCR using broadly reactive primers. Genetically heterogeneous caliciviruses were detected on the majority of farms. Typical noroviruses related to known swine genotypes were present on 20% of the farms. Sapoviruses were detected on 75% of the farms and were the most heterogeneous group. Further characterization of selected strains in their 3′ end parts was carried out for their classification and unveiled possibly new clusters of sapoviruses. No human-like noroviruses or sapoviruses were detected in the present study. GenBank accession numbers of all sequences described in this study are indicated in the figure legends.     

A new species of <Emphasis Type="Italic">Moaciria</Emphasis> (Nematoda,Heterakidae) and other helminths in the red Mawatta frog, <Emphasis Type="Italic">Hylophorbus</Emphasis> cf. <Emphasis Type="Italic">rufescens</Emphasis> (Anura,Microhylidae) from Papua New Guinea

Moaciria moraveci sp. nov. (Nematoda, Heterakidae) from the large intestine of Hylophorbus cf. rufescens from Papua New Guinea is described. Moaciria moraveci sp. nov. represents the 9th species assigned to the genus and the 5th from the Australo-Papuan region. It is distinguished from congeners by the distribution pattern of the caudal papillae of the male, spicule length and vulvar position.     

<Emphasis Type="Italic">Isospora cagasebi</Emphasis> sp. nov. (Apicomplexa,Eimeriidae) from the bananaquit, <Emphasis Type="Italic">Coereba flaveola</Emphasis> of Brazil

Isospora cagasebi sp. nov. (Apicomplexa, Eimeriidae) is reported from a bananaquit, Coereba flaveola from Brazil. Oocysts are sub-spherical, 24.9 × 24.5 (23.0–26.1 × 22.6–25.4), with a smooth, bilayered wall ∼1.4 and mean L:W ratio 1.0; micropyle, oocyst residuum and polar granule are absent. Sporocysts are elongate ovoidal, 18.7 × 11.5 (17.6–19.4 × 10.4–12.3), with both Stieda and substieda bodies and mean L:W ratio 1.6; sporocyst residuum present and sporozoites each with 2 refractiles bodies.     

A new species of genus <Emphasis Type="Italic">Hysterothylacium</Emphasis> Ward et Magath, 1917 (Nematoda,Anisakidae) from <Emphasis Type="Italic">Liparis tanakae</Emphasis> (Scorpaeniformes,Liparidae) from the Yellow Sea,China

A new anisakid nematode, Hysterothylacium liparis sp. nov., is described from the intestine and stomach of the fish, Liparis tanakae (Gilbert et Burke, 1912) (Scorpaeniformes, Liparidae), a fish endemic to the Yellow Sea, China. The new species can be distinguished from the congeners by the absence of lateral alae, the length of the intestinal caecum (1.94–3.35 mm, 58.84–82.47% of oesophageal length), the number and arrangement of the caudal papillae (20–29 precloacal subventral pairs, 1 adcloacal pair and 4 postcloacal pairs), the size of the spicules (1.94–3.74 mm, 4.85–7.30% of body length) and the morphology of the tail tip. This is the first species of adult ascaridoid nematodes to be reported from fishes of the family Liparidae in northern China.     

Use of cross-species <Emphasis Type="Italic">in-situ</Emphasis> hybridization (ZOO-FISH) to assess chromosome abnormalities in day-6 <Emphasis Type="Italic">in-vivo</Emphasis>- or <Emphasis Type="Italic">in-vitro</Emphasis>-produced sheep embryos

Causes of chromosomal differences such as mosaicism between embryos developed in vivo and in vitro may be resolved using animal models to compare embryos generated in vivo with those generated by different production systems. The aims of this study were: (1) to test a ZOO-FISH approach (using bovine painting probes) to detect abnormal chromosome make-up in the sheep embryo model, and (2) to examine the extent of chromosome deviation in sheep embryos derived in vivo and in vitro. Cytogenetic analysis was performed on day 6 in-vivo and in-vitro derived sheep embryos using commercially available bovine chromosome painting probes for sex chromosomes X–Y and autosomes 1–29. A total of 8631 interphase and metaphase nuclei were analyzed from 49 in-vitro-derived and 51 in-vivo-derived embryos. The extent of deviation from normal ovine chromosome make-up was higher (p < 0.05) in in-vitro-produced embryos relative to in-vivo-derived embryos (65.3% vs. 19.6% respectively) mainly due to diploid–polyploid mosaicism. Polyploid cells ranged from 3n to 8n with tetraploids most predominant among non-diploid cells. The proportions of polyploid cells per mixoploid embryo in in-vitro-produced embryos ranged from 1.4% to 30.3%, in contrast to less than 10% among the in-vivo-derived embryos. It was concluded that in-vitro-derived embryos are vulnerable to ploidy change compared to their in-vivo counterparts. The application of ZOO-FISH to domestic animal embryos is an effective approach to study the chromosome complement of species for which DNA probes are unavailable.     

<Emphasis Type="Italic">Anomotaenia barusi</Emphasis> sp. nov. and <Emphasis Type="Italic">A. alata</Emphasis> (Cestoda,Dilepididae) from <Emphasis Type="Italic">Charadrius dubius</Emphasis> (Aves,Charadriiformes) in Slovakia,with comments on related species

During a re-assessment of tapeworm collections from wild birds in Slovakia, two Anomotaenia spp. were recovered from the intestine of the little ringed plover Charadrius dubius Scop., 1786. One of them is described as Anomotaenia barusi sp. nov. The new taxon is distinguished from related congeneric species by the different shape and size of the rostellar hooks, the number of testes and the morphology of male and female reproductive organs. The other species was identified as Anomotaenia alata Spassky et Konovalov, 1969. The validity of this species has formerly been questioned because of its striking morphological similarity to the type-species of the genus, A. microrhyncha (Krabbe, 1869), described from the same host, Philomachus pugnax (L.). Present data revealed differences in the number and measurements of the rostellar hooks, the size of the cirrus-sac, the armament of the cirrus and the presence or absence of setae at the polar ends of the inner egg envelope, which supported the validity of A. alata. The finding of A. alata in C. dubius from Slovakia represents a new host and geographical record.