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The DNA of the prototype strains of ovine adenovirus (OAdV) 1 through 5 was analysed by restriction enzyme (RE) digestion. The RE patterns generated by Hindlll and PstI enzymes were characteristic of the examined strains. OAdV-2 and 3 resembled each other the most, and their EcoRI and Hindlll patterns seemed to be identical. Considering the number of comigrating fragments, serotypes OAdV-2, 3, 4 and 5 looked more closely related to each other than to OAdV-1. This finding was strengthened by Southern blot hybridisations probed with random Hindlll clones of OAdV-3. The estimated genome size of the examined OAdV types ranged between 31.9 and 32.8 kilobase pairs. The results supported the new genus classification of OAdVs.

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To explore the diversity of some DNA viruses in reptiles, a continuous screening is going on, in our laboratory, by PCR using different consensus primers designed for the detection of the most conserved genome regions of adeno-, herpes- and parvoviruses. The test material consists essentially of dead specimens collected randomly from private pet owners, local pet shops, or at occasional exotic pet fairs. Here we report the partial sequence of a putative novel parvovirus obtained from a dead checkerboard worm lizard (Trogonophis wiegmanni) that had been wild-caught in its native habitat. An in-house-developed PCR with consensus primers targeting the gene of the parvoviral capsid protein was used. Other PCRs, intended to detect certain large DNA viruses, remained negative. The sequence of the PCR product indicated the presence of a hitherto unknown parvovirus in the internal organs of the checkerboard worm lizard. In phylogeny reconstruction, the novel sequence clustered with the members of the Dependovirus genus of the Parvoririnae subfamily, closest to the branch of snake adeno-associated virus. Since we could not demonstrate the presence of a potential helper virus, the putative amphisbaenian parvovirus supposedly can replicate autonomously. This is the first virus infection ever detected in any members of the suborder Amphisbaenia, and only the third parvoviral sequence obtained from any reptilian host.

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Adenoviruses are frequent infectious agents in different poultry species. The traditional, serological typing of new isolates by virus neutralisation tests is now in transition to be replaced by PCR and sequencing. The first PCRs, recommended for the detection of adenoviruses, had been designed to target the gene of the major capsid protein, the hexon. In birds, members of three different genera of the family Adenoviridae may occur. Accordingly, three specific hexon PCRs had to be elaborated for the detection of adenoviruses in poultry. A significantly more sensitive PCR, targeting the viral DNA-dependent DNA polymerase gene, has been described recently. This method proved to be an efficient alternative for the general detection of adenoviruses irrespective of their genus affiliation. Fowl adenoviruses (FAdVs), isolated from chicken to date, comprise twelve serotypes classified into five virus species (FAdV-A to E). The polymerase gene sequence has been determined yet only from three FAdV types representing three species. In the present work, the panel of polymerase gene sequences was completed with those of the rest of FAdVs. The newly determined sequences will facilitate the identification of new FAdV isolates as an existing species or as a putative new FAdV. Once the polymerase sequence is known, more specific PCRs for the amplification of the hexon and other genes can be designed and performed according to the preliminary species classification.

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Abstract

By a broad-range PCR, we detected a novel herpesvirus (HV) in the specimen of a wels catfish (Silurus glanis) presenting disseminated, carp pox-like dermal lesions all over its body. The sequence analysis of the 463-bp PCR product from the viral DNA polymerase gene indicated the presence of a hitherto unknown virus, a putative member of the family Alloherpesviridae in the sample. Another PCR, targeting the terminase gene of fish HVs, provided an additional genomic fragment of over 1,000 bp. Surprisingly, the sequence of a co-amplified, off-target PCR product revealed its origin from a putative gene homologous to ORF87 and ORF45 of cyprinid HVs and anguillid herpesvirus 1 (AngHV-1), respectively. With specific primers, designed according to the genomic maps of the cyprinid and anguillid HVs, a genomic fragment of 15 kb was also amplified and sequenced by primer walking. In phylogeny inferences, based on several genes, the putative wels catfish HV clustered closest to various cyprinid HVs or to AngHV-1. The novel virus, named as silurid herpesvirus 2, represents a distinct species in the genus Cyprinivirus. However, its association with the skin disease remains unclear.

Open access

The negative samples of a collection, established originally for seeking new adeno- and herpesviruses in lower vertebrates, were screened for the pres-ence of circoviruses by a consensus nested PCR targeting the gene coding for the replication-associated protein. Six fish samples representing five species, namely asp (Aspius aspius), roach (Rutilus rutilus), common bream (Abramis brama), round goby (Neogobius melanostomus) and monkey goby (Neogobius fluviatilis), as well as three frog samples were found positive for circoviral DNA. Sequence analysis of the amplicons indicated the presence of three novel putative circo-like viruses and a circovirus in Hungarian fishes and one novel circovirus in a common toad (Bufo bufo), and another one in a dead and an alive specimen of green tree frog (Litoria caerulea), respectively. In phylogeny reconstruction, the putative bream circovirus clustered together with circoviruses discovered in other cyprinid fishes recently. Three other piscine circoviral sequences appeared closest to sequences derived from different environmental samples. Surprisingly, the nucleotide sequence derived from two fish samples (a bream and a monkey goby) proved to be from porcine circovirus 2 (PCV2), almost identical to a sequence detected in Sweden previously. This is the first report on the detection of PCV2 in fish and circoviral DNA in amphibian hosts.

Open access
Orvosi Hetilap
Authors:
Mária Matuz
,
Ria Benkő
,
Edit Hajdú
,
Réka Viola
, and
Gyöngyvér Soós

Bevezetés: A bakteriális rezisztencia visszaszorítására az antibiotikumok átgondolt alkalmazása lehetséges beavatkozási pont. Célkitűzés: A hazai ambuláns antibiotikum-alkalmazás gyakorlatának nemzetközi minőségi indikátorokon keresztül történő bemutatása. Módszer: Az 1996–2010 közötti időszakra vonatkozó nyers, nagykereskedői eladásokon alapuló, ambuláns szisztémás antibiotikum-felhasználási adatokat az Egészségügyi Világszervezet 2010. évi napi terápiás dózisai (defined daily dose – DDD) figyelembevételével DDD/1000 fő/nap egységben adták meg. Az antibiotikum-felhasználás értékelésére a nemzetközileg kidolgozott és validált minőségi indikátorokat alkalmazták. Eredmények: A hazai ambuláns antibiotikum-alkalmazás a vizsgálat során mennyiségileg kiegyenlített (18,0±1,8 DDD/1000 fő/nap) volt, szerkezetében azonban jelentős változások történtek. A széles versus szűk spektrumú béta-laktámok és makrolidek felhasználási aránya többszörösére nőtt (1996: 2,2 vs. 2010: 15,8), a fluorokinolonok felhasználása megháromszorozódott. A vizsgált tíz minőségi indikátor közül Magyarország három indikátor esetében az európai elithez, négy, valamint három esetében a gyengébb, illetve leggyengébb országok közé tartozott. Következtetés: A hazai ambuláns antibiotikum-felhasználás mennyiségileg a skandináv, az összetétel tekintetében a déli országokhoz hasonlít. Orv. Hetil., 2013, 154, 947–956.

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Abstract

Extensive use of carbapenems may lead to selection pressure for Stenotrophomonas maltophilia (SM) in hospital environments. The aim of our study was to assess the possible association between systemic antibiotic use and the incidence of SM. A retrospective, observational study was carried out in a tertiary-care hospital in Hungary, between January 1st 2010 and December 31st 2019. Incidence-density for SM and SM resistant to trimethoprim-sulfamethoxazole (SXT) was standardized for 1000 patient-days, while systemic antibiotic use was expressed as defined daily doses (DDDs) per 100 patient-days. Mean incidence density for SM infections was 0.42/1000 patient-days; 11.08% were were resistant to SXT, the mean incidence density for SXT-resistant SM was 0.047/1000 patient-days. Consumption rate for colistin, glycopeptides and carbapenems increased by 258.82, 278.94 and 372.72% from 2010 to 2019, respectively. Strong and significant positive correlations were observed with the consumption of carbapenems (r: 0.8759; P < 0.001 and r: 0.8968; P < 0.001), SXT (r: 0.7552; P = 0.011 and r: 0.7004; P = 0.024), and glycopeptides (r: 0.7542; P = 0.012 and r: 0.8138; P < 0.001) with SM and SXT-resistant SM incidence-density/1000 patient-days, respectively. Implementation of institutional carbapenem-sparing strategies are critical in preserving these life-saving drugs, and may affect the microbial spectrum of infections in clinical settings.

Open access

The full sequence of the fiber gene and partial sequence of the putative 17 kD protein gene of bovine adenovirus-2 (BAdV-2) were determined. The size of the fiber gene of BAdV-2 proved to be 561 amino acids, of which the amino acids 37 to 385 form a typical shaft domain of 22 repetitive motifs. On the complementary strand, a gene homologous to the 17 kD protein coded in the E4 region of several human adenoviruses was found. The sequence analysis seems to confirm the presence of an intron in the sequenced part of the E4 region.

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Adenoviral nucleic acid was detected by polymerase chain reaction (PCR) in pharyngeal and rectal swab samples of a cat seropositive for adenovirus and suffering from transient hepatic failure. The samples were taken at a one-year interval, and both faecal samples as well as the second pharyngeal sample were positive in PCR performed with general adenovirus primers. The size of the amplified products corresponded to that of the positive control. The identity of the amplicons was also confirmed by DNA sequencing. The 301 bp long hexon gene fragment was very similar to but distinguishable from the corresponding hexon sequence of human adenovirus type 2. This result suggests the possibility of persistent carrier status and shedding of adenovirus in cats.

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