Novel picornavirus in domesticated common quail (<Emphasis Type="Italic">Coturnix coturnix</Emphasis>) in Hungary

This study reports the detection of a novel picornavirus in domesticated common quail (Coturnix coturnix) in Hungary. The 8159-nucleotide (nt)-long RNA genome of this virus, named quail picornavirus (QPV1-HUN/2010; JN674502), shows only 43%, 39% and 47% amino acid (aa) identity in the P1 (857 aa), P2 (458 aa) and P3 (777 aa) coding regions respectively, to the closest reference, avian sapelovirus. The 5′UTR contains a variant type IV IRES with a 20-nt-long apical “8”-like structure that is conserved in avian-origin and seal picornaviruses. The 390-aa-long L protein is cysteine rich and encodes two copies of a 34-aa-long repeat motif. Quail picornavirus represents a novel picornavirus species and perhaps a novel genus.     

Complete genomic sequence analysis of a highly virulent isolate revealed a novel strain of <Emphasis Type="Italic">Sugarcane mosaic virus</Emphasis>

Sugarcane mosaic virus (SCMV) is the most prevalent virus causing maize dwarf mosaic disease in northern China. A SCMV isolate, BD8, was obtained from the maize showing dwarf and mosaic symptoms in Baoding, China. The complete genomic sequence of BD8 is 9,576 nucleotides (nt) excluding the poly(A) tail. It contains one single open reading frame of 9,192 nt and encodes a large polyprotein of 3,063 amino acids (aa), flanked by a 5′-untranslated region (UTR) of 148 nt and a 3′-UTR of 236 nt. The entire genomic sequence of BD8 shares identities of 79.1–80.8% with those of other 13 SCMV isolates available in the GenBank at nt level, while their CP genes share identities of 76.9–82.6 and 82.8–86.9% at nt and aa levels, respectively. Phylogenetic analysis of the complete genomic sequences reveals that SCMV can be clustered to four groups: group I includes isolates from maize, group II consists of isolates from sugarcane or maize, groups III and IV contain single isolate of AU-A (AJ278405) and BD8, respectively. Thus, BD8 represents a new strain of SCMV. Furthermore analysis of the CP gene sequences of more isolates shows that BD8 is clustered to a group with the isolates from Thailand and Vietnam, which implies that isolates of this strain have been distributed in South Asia. In the greenhouse, BD8 can cause severe symptoms in all the 12 maize varieties tested with high incidence, indicating that BD8 is highly virulent.     

Isolation,characterization and genome sequence of a birnavirus strain from flounder <Emphasis Type="Italic">Paralichthys </Emphasis><Emphasis Type="Italic">olivaceus</Emphasis> in China

A birnavirus strain, Paralichthys olivaceus birnavirus (POBV), was isolated and characterized from cultured flounder in China, and its complete genomic sequence was subsequently determined. The virus could induce cytopathic effects (CPE) in four of seven fish cell lines and was resistant to chloroform, 5-iodo-2′-deoxyuridine, acid and alkaline pH, and heat treatment. Purified virus particles had a typical icosahedral shape, with a diameter of approximately 55–60 nm. The genomic segments A and B of POBV were 3,091 and 2,780 bp in length and shared many of the features of the members of the family Birnaviridae. Segment A contained two partially overlapping ORFs encoding a polyprotein, pVP2-VP4-VP3, and a nonstructural protein, VP5, while segment B had only one ORF encoding for the VP1, a viral RNA-dependent RNA polymerase (RdRp). This is the first report about a birnavirus strain from a new non-salmonid host in China and its complete genome sequence. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users. The complete genome sequences of POBV reported here have been deposited in the GenBank database under accession no EU161285 and EU161286, respectively.     

High sequence conservation among <Emphasis Type="Italic">Odotoglossum ringspot virus</Emphasis> isolates from orchids

The variability in the nucleotide (nt) and amino acid (aa) sequences of the coat protein (CP) of Odontoglossum ringspot virus (ORSV), which naturally infects orchids worldwide, was investigated. The CP genes of 48 ORSV isolates originating from different locations in Korea were amplified using RT-PCR and sequenced. The encoded CP consists of 158 aa. The CP sequences of the Korean isolates were compared at the nt and aa levels with those of the previously published ORSV isolates originating from different countries. The Korean isolates share 94.8–100% and 92.4–100% CP identity to ORSV isolates from other countries at the nt and aa levels, respectively. No particular region of variability could be found in either sequence of the viruses. In the deduced aa sequence, the N-terminal region was more conserved than the C-terminal region in ORSV. The phylogenetic tree analysis and recombination analysis revealed that there was no distinct grouping between geographic locations and sequence identity, and nor distinct intra-specific recombination events among ORSV isolates.     

A new potyvirus first isolated and identified from <Emphasis Type="Italic">Angelica sinensis</Emphasis>

A filamentous virus was isolated in Angelica sinensis (Angelica sinensis (Oliv.) Diels) which shows mosaic symptoms on leaves in Minxian, Gansu province, China. According to morphology and molecular biology properties, this virus, which has a flexuous rod-shaped particle about 750 nm in length and 12 nm in width, was assigned to the genus Potyvirus, family Potyviridae. Its coat protein (CP) shows high similarity with six other potyviruses by analysis of peptide mass fingerprinting (PMF). The 919 bp nucleotides of 3′ terminal covering partial CP gene and 3′-untranslated region was amplified by RT-PCR using degenerate primers which were designed according to the result of PMF. In sequence comparisons and phylogenetic analysis, the new isolate was found to be closely related to Japanese hornwort mosaic virus (JHMV), Konjak mosaic virus (KoMV), and Zantedeschia mosaic virus (ZaMV). The most closely related virus is JHMV03 (AB251346), with 96.59% aa and 87.60% nt identity to the isolate. All results suggest the presence of a new member of potyvirus, tentatively named Dang Gui strain of Japanese hornwort mosaic virus (JHMV-DG*). In our research the antiserum against the CP of JHMV-DG had also been prepared. To our knowledge, it is the first time that a potyvirus has been isolated and identified in Angelica sinensis. *Nucleotide sequence date reported appear in GenBank with the accession number EF157843.     

A virus related to <Emphasis Type="Italic">Soybean mosaic virus</Emphasis> from <Emphasis Type="Italic">Pinellia ternata</Emphasis> in China and its comparison with local soybean SMV isolates

Summary. A potyvirus isolated from Pinellia ternata in China was characterised and shown to be related to Soybean mosaic virus (SMV). The virus was pathogenic on P. ternata and some soybean cultivars, whereas the local soybean SMV isolate HH5 did not infect P. ternata. Western blot experiments demonstrated a serological relationship between the virus from Pinellia, SMV and Watermelon mosaic virus (WMV). The complete nucleotide sequences of the Pinellia virus (isolate P-1, 9735nt) and of the Chinese soybean SMV isolates HH5 (9585nt) and HZ (9588nt) were determined. A 1733nt sequence at the 3-terminus of a second isolate from Pinellia (isolate P-2) was also determined. The predicted polyprotein of isolate P-1 has 83% amino acid (aa) identity with those of published SMV sequences. In many parts of the genome, aa identity was about 90% but it was much lower in the P1 protein region (24–29%), where it more closely resembled Dasheen mosaic virus (62%). The partial sequence of isolate P-2 had 91%nt identity to P-1 and both isolates resembled a recent sequence in the public databases (AF469171) wrongly named Zantedeschia mosaic virus. The two complete SMV soybean sequences had 93–95%nt identity with those of the previously sequenced isolates and >97% amino acid identity. Phylogenetic analysis and comparisons of coat proteins suggest that the Pinellia, WMV and SMV potyviruses should probably be treated as strains of the same species.     

The combination of <Emphasis Type="Italic">ApoCIII</Emphasis>, <Emphasis Type="Italic">hepatic lipase</Emphasis> and <Emphasis Type="Italic">hormono sensitive lipase</Emphasis> gene polymorphisms suggests an association with susceptibility to gestational hypertension

Dyslipidemia and insulin resistance contribute to the endothelial cell dysfunction in hypertensive disorders of pregnancy (HDP) and increase the long-term risk of cardiovascular disease (CVD). The genes linking susceptibility to gestational hypertension (GH) and/or preeclampsia (PE) to the long-term risk of CVD are still unknown. We evaluated the potential association between 14 polymorphisms from six genes involved in lipid metabolism and insulin action and the risk of HDP: namely the lipoprotein lipase (LPL), hepatic lipase (LIPC), hormone sensitive lipase (LIPE), cholesteryl ester transfer protein (CETP), ApoCIII and ApoE gene polymorphisms. Overall, 169 women with HDP [proteinuria (PE) and gestational hypertension without proteinuria (GH)] and 169 controls matched for age and year of delivery were genotyped. Homozygosity of the −514T allele of the −514C > T polymorphism (LIPC gene) decreased the risk of GH (OR = 0.17, CI₉₅: 0.02–0.76), while there were more −60G carriers of the −60C > G LIPE gene polymorphism (OR = 3.51, CI₉₅:1.02–12.10) among GH cases, but not in PE cases. The common ApoCIII two-locus −482CC/3238CC genotype was lower in women with GH compared with controls (OR = 0.53, CI₉₅: 0.3–0.9). The combined frequency of at-risk genotypes was higher in cases of GH compared with controls [one at-risk genotype: OR = 3.38 (95% CI: 0.48–41.8); two or more at-risk genotypes: OR = 7.14 (95% CI: 1.21–92.3, P = 0.01)], suggesting a gene-dose effect. We conclude that the combined effect of LIPC, LIPE and ApoCIII gene polymorphisms may increase the likelihood of GH, but seemingly not of PE.     

A new species of <Emphasis Type="Italic">Hysterothylacium</Emphasis> (Ascaridoidea,Anisakidae) parasitic in <Emphasis Type="Italic">Zenopsis conchifer</Emphasis> (Zeiformes,Zeidae) from Argentinean waters

A new species of parasitic nematode, Hysterothylacium spirale sp. nov. (Ascaridoidea, Anisakidae), is described based on specimens collected from the intestine and pyloric caeca of the silvery john dory Zenopsis conchifer (Lowe) (Zeiformes, Zeidae), from the Argentinean Shelf (35°05′–40°46′S, 53°03′–58°07′W). Among the 66 valid species described so far in the genus, the new species most closely resembles H. zenis (Baylis, 1929). Both species, apparently specific for fishes of the family Zeidae, share the shape of the dorsal lip, the long expanded lateral alae originating from subventral interlabia and the ornamentation of the tail tip, as well as general morphometry. The combination of these shared features distinguishes both species from all congeners so far known. However, the new species is distinguishable from H. zenis by having shorter interlabia, and consequently the lateral alae originating more posteriorly, shorter spicules, a smaller number of postcloacal papillae and the presence of two pairs of double postcloacal papillae.     

<Emphasis Type="Italic">Melandrium yellow fleck bromovirus</Emphasis> infects <Emphasis Type="Italic">Arabidopsis thaliana</Emphasis> and has genomic RNA sequence characteristics that are unique among bromoviruses

Melandrium yellow fleck bromovirus (MYFV) systemically infected Arabidopsis thaliana, although the susceptibility of several A. thaliana accessions to MYFV differed from their susceptibility to the other two bromoviruses infecting A. thaliana. We constructed full-length cDNA clones of MYFV genomic RNAs 1, 2, and 3 and determined their complete nucleotide sequences. Similar to Broad bean mottle bromovirus, (1) the 5′-terminal nucleotide of the MYFV genomic RNAs was adenine, and (2) the “D-arm” was absent from the tRNA-like structure in the 3′ untranslated regions (UTRs) of MYFV RNAs. As unique characteristics, MYFV RNA3 lacked the poly(A) tract in the intercistronic region and contained a directly repeated sequence of about 200 nucleotides and polypyrimidine tracts of heterogeneous lengths in the 5′ UTR. Co-infection experiments using RNA3 clones with or without the duplicated sequence demonstrated that the duplication contributed to the competitive fitness of the virus in Nicotiana benthamiana.     

Copper tolerance of <Emphasis Type="Italic">Saccharomyces cerevisiae</Emphasis> nonsense-mediated mRNA decay mutants

The eukaryotic nonsense-mediated mRNA (NMD) is a specialized pathway that leads to the recognition and rapid degradation of mRNAs with premature termination codons, and importantly some natural mRNAs as well. Natural mRNAs with atypically long 3′-untranslated regions (UTRs) are degraded by NMD in Saccharomyces cerevisiae. A number of S. cerevisiae mRNAs undergo alternative 3′-end processing producing mRNA isoforms that differ in their 3′-UTR lengths. Some of these alternatively 3′-end processed mRNA isoforms have atypically long 3′-UTRs and would be likely targets for NMD-mediated degradation. Here, we investigated the role NMD plays in the regulation of expression of CTR2, which encodes a vacuolar membrane copper transporter. CTR2 pre-mRNA undergoes alternative 3′-end processing to produce two mRNA isoforms with 300-nt and 2-kb 3′-UTRs. We show that both CTR2 mRNA isoforms are differentially regulated by NMD. The regulation of CTR2 mRNA by NMD has physiological consequences, since nmd mutants are more tolerant to toxic levels of copper relative to wild-type yeast cells and the copper tolerance of nmd mutants is dependent on the presence of CTR2.     

Prognostic value of procalcitonin in <Emphasis Type="Italic">Legionella</Emphasis> pneumonia

The diagnostic reliability and prognostic implications of procalcitonin (PCT) (ng/ml) on admission in patients with community-acquired pneumonia (CAP) due to Legionella pneumophila are unknown. We retrospectively analysed PCT values in 29 patients with microbiologically proven Legionella-CAP admitted to the University Hospital Basel, Switzerland, between 2002 and 2007 and compared them to other markers of infection, namely, C-reactive protein (CRP) (mg/l) and leukocyte count (10⁹/l), and two prognostic severity assessment scores (PSI and CURB65). Laboratory analysis demonstrated that PCT values on admission were >0.1 in over 93%, >0.25 in over 86%, and >0.5 in over 82% of patients with Legionella-CAP. Patients with adverse medical outcomes (59%, n = 17) including need for ICU admission (55%, n = 16) and/or inhospital mortality (14%, n = 4) had significantly higher median PCT values on admission (4.27 [IQR 2.46–9.48] vs 0.97 [IQR 0.29–2.44], p = 0.01), while the PSI (124 [IQR 81–147] vs 94 [IQR 75–116], p = 0.19), the CURB65 (2 [IQR 1–2] vs 1 [1–3], p = 0.47), CRP values (282 [IQR 218–343], p = 0.28 vs 201 [IQR 147–279], p = 0.28), and leukocyte counts (12 [IQR 10–21] vs 12 [IQR 9–15], p = 0.58) were similar. In receiver operating curves, PCT concentrations on admission had a higher prognostic accuracy to predict adverse outcomes (AUC 0.78 [95%CI 0.61–96]) as compared to the PSI (0.64 [95%CI 0.43–0.86], p = 0.23), the CURB65 (0.58 [95%CI 0.36–0.79], p = 0.21), CRP (0.61 [95%CI 0.39–0.84], p = 0.19), and leukocyte count (0.57 [95%CI 0.35–0.78], p = 0.12). Kaplan-Meier curves demonstrated that patients with initial PCT values above the optimal cut-off of 1.5 had a significantly higher risk of death and/or ICU admission (log rank p = 0.003) during the hospital stay. In patients with CAP due to Legionella, PCT levels on admission might be an interesting predictor for adverse medical outcomes. Jeannine Haeuptle, Roya Zaborsky, and Rico Fiumefreddo contributed equally to this article.     

Occurrence of <Emphasis Type="Italic">Cryptosporidium parvum</Emphasis> and <Emphasis Type="Italic">Giardia duodenalis</Emphasis> in healthy adult domestic ruminants

To determine the prevalence and intensity of infection of Cryptosporidium spp. and Giardia duodenalis in healthy adult domestic ruminants, faecal samples were collected from 379 cattle of between 3 and 13 years old, 446 sheep and 116 goats selected at random from 60 dairy farms and 38 and 20 herds, respectively, in Galicia (NW Spain). Cryptosporidium spp. oocysts were detected in 32 cows (8.4%), 24 sheep (5.3%) and in nine goats (7.7%) from, respectively, 48.3% of the farms and 34.2 and 30.0% of the herds. The intensity of infection in cows ranged between 25 and 5,924 oocysts per gram of faeces (OPG), whereas in sheep and goats, the number of oocysts shed ranged from 8–515 OPG and from 17–782 OPG, respectively. Parasitization by Cryptosporidium spp. was significantly higher (P < 0.05) in cows than in sheep and goats. G. duodenalis cysts were identified in 101 cows (26.6%), 86 sheep (19.2%) and 23 goats (19.8%) from, respectively, 96.6% of the farms and 92.1 and 90% of the herds. The number of cysts shed by cows ranged between 15 and 3,042 cyst per gram of faeces (CPG), whereas the intensity of infection in sheep and goats ranged from 16–3010 CPG and from 15–1845 CPG, respectively, and was significantly lower (P < 0.05) than in cows and sheep. The number of Cryptosporidium spp. oocysts isolated from sheep and goats was insufficient for successful polymerase chain reaction analysis. Nevertheless, gene sequence analysis of the hsp70 and 18SrRNA genes of Cryptosporidium revealed the presence of only C. parvum in faecal samples from cows. Genotyping studies of the β-giardin and glutamate dehydrogenase genes of G. duodenalis revealed mainly assemblage E of Giardia in cows, sheep and goat faecal samples. Assemblage B of G. duodenalis was also detected in one sheep sample. These animals should be considered as a possible source of cryptosporidiosis and giardiosis, thereby maintaining the infections on farms and in herds.     

Molecular identification of <Emphasis Type="Italic">Trichuris vulpis</Emphasis> and <Emphasis Type="Italic">Trichuris suis</Emphasis> isolated from different hosts

Trichuris suis was isolated from the cecum of two different hosts (Sus scrofa domestica—swine and Sus scrofa scrofa—wild boar) and Trichuris vulpis from dogs in Sevilla, Spain. Genomic DNA was isolated and internal transcribed spacers (ITS)1-5.8S-ITS2 segment from the ribosomal DNA (rDNA) was amplified and sequenced using polymerase chain reaction techniques. The sequence of T. suis from both hosts was 1,396 bp in length while that of T. vulpis was 1,044 bp. ITS1 of both populations isolated of T. suis was 661 nucleotides in length, while the ITS2 was 534 nucleotides in length. Furthermore, the ITS1 of T. vulpis was 410 nucleotides in length, while the ITS2 was 433 nucleotides in length. One hundred fifty-four nucleotides were observed along the 5.8S gene of T. suis and T. vulpis. Intraindividual and intraspecific variations were detected in the rDNA of both species. The presence of microsatellites was observed in all the individuals assayed. Sequence analysis of the ITSs and the 5.8S gene has demonstrated no sequence differences between T. suis isolated from both hosts (S. scrofa domestica—swine and S. scrofa scrofa—wild boar). Nevertheless, clear differences were detected between the ITS1 and ITS2 of T. suis and T. vulpis. Furthermore, a comparative molecular analysis between both species and the previously published ITS1-5.8S-ITS2 sequence data of Trichuris ovis, Trichuris leporis, Trichuris muris, Trichuris arvicolae, and Trichuris skrjabini was carried out. A common homology zone was detected in the ITS1 sequence of all species of trichurids.     

Genetic interactions model among <Emphasis Type="Italic">Eotaxin</Emphasis> gene polymorphisms in asthma

Eotaxin family (Eotaxin 1,2 and 3) recruits and activates CCR3-bearing cells such as eosinophil, mast cells, and Th2 lymphocytes that play a major role in allergic disorders. We examined the polygenetic effects of the Eotaxin gene family in a Korean population. Gene–gene interactions were tested using a multistep approach with multifactor dimensionality reduction (MDR) method between asthmatics and normal controls. The overall best MDR model of the main effect single nucleotide polymorphisms (SNPs) included EOT2 + 1272A > G and EOT3 + 77C > T (model 1) [testing accuracy 0.597, cross-validation consistency (CVC) 10/10, P < 0.001]. The overall best MDR model of the SNPs with no main effects included EOT2 + 304C > A, EOT3 + 716A > G, and EOT3 + 1579G > A (model 2) (testing accuracy 0.616, CVC 10/10, P < 0.001). Model 3 was obtained by including the MDR variables for models 1 and 2. This new composite model predicted asthma with better accuracy than either model 1 or model 2 (testing accuracy 0.643, CVC 10/10, P < 0.001). The detection of statistical interaction models is one evidence of gene–gene interactions among Eotaxin genes, and this interaction is thought to influence the development of asthma. Although the models are limited to determining statistical interactions within a population, they may be useful for identifying groups at high risk of developing asthma.     

Analysis and characterization of the complete genome of a member of a new species of kobuvirus associated with swine

A virus belonging to a new species in the genus Kobuvirus, family Picornaviridae, was first isolated in 2008 from apparently healthy pigs in Hungary and China. We report the complete genome sequence and the genetic organization of the novel porcine kobuvirus strain Y-1-CHI, which was identified in China. The RNA genome of strain Y-1-CHI contains 8210 nucleotides (nt) and has an organization similar to that of other picornaviruses. The full-length nucleotide sequence of Y-1-CHI was 88.62%, 58.66%, and 48.86% identical to those of S-1-HUN, U-1, and Aichi virus, respectively. No positive results were found in 454 stool samples from children with acute gastroenteritis. Dendrograms indicated that Y-1-CHI and S-1-HUN are most closely related to each other and belong to the same species. Our results suggest that members of this novel species have the typical genome characteristics of members of the genus Kobuvirus and may be distributed globally in swine.     

<Emphasis Type="Italic">Sarcocystis</Emphasis> sp. from the goldeneye (<Emphasis Type="Italic">Bucephala clangula</Emphasis>) and the mallard (<Emphasis Type="Italic">Anas platyrhynchos</Emphasis>): cyst morphology and ribosomal DNA analysis

By light microscopy, cysts of Sarcocystis sp. (cyst type I) from the goldeneye (Bucephala clangula) seemed filamentous with a smooth and thin (<1 μm) cyst wall. Ultrastructurally, the cyst wall surface was irregular with minute undulations of the primary cyst wall. These sarcocysts had type-1 cyst wall. Cystozoites were banana-shaped and measured 7.0–8.5 μm in length. By light microscopy, cysts of Sarcocystis sp. (cyst type II) from the mallard (Anas platyrhynchos) were ribbon-shaped, very long, and thin. On the surface of the wall (up to 1.5 μm), they had palisade-like villar protrusions closely crowded together. Electron micrographs showed villar protrusions (up to 1.3 μm in length) different in size and shape. The latter had short microprojections especially obvious in the oblique sections. Cystozoites were slightly bent with blunt ends, broader at one end, and measured 13.0–16.1 × 1.8–2.5 μm. Phylogenetic analysis based on the comparison of partial 28S rRNR gene sequences of Sarcocystis sp. (cyst type II) derived from the mallard, Sarcocystis sp. (cyst type I) and Sarcocystis sp. (cyst type III) both derived from the white-fronted goose (Anser albifrons) suggested that these sequences belonged to separate Sarcocystis species.     

<Emphasis Type="Italic">TPH2</Emphasis> and <Emphasis Type="Italic">TPH1</Emphasis>: Association of Variants and Interactions with Heroin Addiction

The tryptophan hydroxylase 2 gene (TPH2) was resequenced at the 5′ upstream, coding, and 3′ downstream regions, including all 11 exons in 185 subjects. Twenty-three novel and 14 known variants were identified. In a cohort of 583 consecutively ascertained subjects, including normal volunteers and those with specific addictive diseases, six common TPH2 and one TPH1 variant were genotyped. Allele frequencies of three TPH2 variants and the TPH1 variant varied significantly among the four ethnic groups within the control subjects. Of these subjects, 385 who met heroin addiction or control criteria and were of Caucasian, African-American, or Hispanic ethnicity were examined for potential association with vulnerability to develop heroin addiction. At the two locus genotype level in Hispanics, the TPH1 rs1799913 variant was found to significantly interact with the TPH2 rs7963720 variant and heroin addiction (P = 0.022), and with the TPH2 rs4290270 variant and heroin addiction (P = 0.011). In the African-American group, a significant association of a specific TPH2 haplotype with heroin addiction also was found (SNPHAP, P = 0.004; PHASE P = 0.036). Edited by Tatiana Foroud.     

Nucleotide sequences of four segments of chrysovirus in Korean <Emphasis Type="Italic">Cryphonectria nitschkei</Emphasis> BS122 strain

The near full-length genome consisting of four segments of dsRNA from a chrysovirus infecting Korean Cryphonectria nitschkei BS122 strain (CnV1-BS122) was sequenced. The open reading frames of segments 1, 2, 3, and 4 were 2,889, 2,721, 2,475, and 2,232 nucleotides (nt) in length, respectively. Sequence analysis and homology searches of the amino acid sequences deduced from the ORFs of each segment revealed that segments 1, 2, 3, and 4 encoded RNA-dependent RNA polymerase, capsid protein, a putative cysteine protease, and replication-associated protein, respectively. The entire 5′ ends of segments 1, 2, and 4 were 82, 242, and 698 nt in length, respectively; the sequence of the 5′ end of segment 3 was not determined because of difficulty in amplification. The entire 3′ end of segment 3 was 77 nt in length. Partial amplification of the 3′ ends of segments 1, 2, and 4 yielded amplimers of 7, 17, and 30 nt, respectively.     

Occurrence of cystacanths of <Emphasis Type="Italic">Plagiorhynchus cylindraceus</Emphasis> (Acanthocephala) in the terrestrial isopods <Emphasis Type="Italic">Trachelipus squamuliger</Emphasis> and <Emphasis Type="Italic">Armadillidium vulgare</Emphasis> (Oniscidea) in Bulgaria

In total, 2097 individuals of Trachelipus squamuliger and 20 individuals of Armadillidium vulgare from four habitats (three woodland sites and one pasture) in the region of Stara Zagora, Bulgaria, were examined for the presence of cystacanths of Plagiorhynchus cylindraceus, a common acanthocephalan parasite of passerine birds. In T. squamuliger from woodland habitats, cystacanths were found with prevalence 4.0–9.3%, intensity 1–5 (mean 1.22–1.57) and mean abundance 0.057–0.113. No significant differences were observed between infections in males and females of T. squamuliger. None of the T. squamuliger individuals from the pasture examined was infected. Out of 48 infected females of T. squamuliger, only one had developed eggs (in agreement with previous studies revealing the negative effect of the cystacanths on the development of female gonads of woodlice). One individual of A. vulgare was infected with a single cystacanth. The occurrence of P. cylindraceus in T. squamuliger is a new host record.