RESUMO
The Eurasian beaver (Castor fiber), native to Hungary, faced local extinction in 1865 and was successfully reintroduced between mid-1980s and 2008. Despite screening programs focusing on animal health during reintroduction in other countries, information about viruses in the Hungarian beaver population remains limited. Polyomaviruses (PyVs) have been identified in various rodents, and have been detected just recently in beavers by us. In this paper we present the full genome analysis of the first PyV detected in Eurasian beaver. The novel PyV was discovered in the kidney tissues of two specimens. The genome is 5244 bp, and contains four genes. Small T-antigen (STAg) and alternative large T ORF (ALTO) genes are directly fused together forming the middle T-antigen (MTAg). VP3 is absent from the genome. Its large T-antigen (LTAg) coding sequence exhibited over 15% genetic divergence from known PyVs, supporting its classification into a new species within the genus Alphapolyomavirus, suggesting to be named Alphapolyomavirus castoris. Phylogenetic analysis, based on the LTAg gene showed, that the beaver PyV forms a distinct clade with primate PyVs within the genus Alphapolyomavirus, separate from other rodent PyVs. Phylogenetic study of the VP1 gene however showed this virus to belong in a distinct clade with the same primate PyVs, and additionally PyVs from rodents and a myocastor, which suggest host virus co-evolution. The virus detection of the euthanized beavers suggests an apathogenic persistent infections. The aquatic lifestyle of beavers may influence virus transmission, warranting further exploration of undiscovered viruses in beavers.
Assuntos
Genoma Viral , Filogenia , Infecções por Polyomavirus , Polyomavirus , Roedores , Animais , Polyomavirus/genética , Polyomavirus/classificação , Polyomavirus/isolamento & purificação , Roedores/virologia , Infecções por Polyomavirus/virologia , Infecções por Polyomavirus/veterinária , Hungria , Sequenciamento Completo do Genoma , Doenças dos Roedores/virologia , Doenças dos Roedores/epidemiologiaRESUMO
Polyomaviruses (PyVs) are known to infect a diverse range of vertebrate host species. We report the discovery of PyVs in vesper bats (family Vespertilionidae) from sampling in Central Europe. Seven partial VP1 sequences from different PyVs were detected in samples originating from six distinct vesper bat species. Using a methodology based on conserved segments within the major capsid virus protein 1 (VP1) among known PyVs, the complete genomes of two different novel bat PyVs were determined. The genetic distances of the large T antigen coding sequences from these PyVs compared to previously-described bat PyVs exceeded 15% meriting classification as representatives of two novel PyV species: Alphapolyomavirus epserotinus and Alphapolyomavirus myodaubentonii. Phylogenetic analysis revealed that both belong to the genus Alphapolyomavirus and clustered together with high confidence in clades including other bat alphapolyomaviruses reported from China, South America and Africa. In silico protein modeling of the VP1 subunits and capsid pentamers, and electrostatic surface potential comparison of the pentamers showed significant differences between the reference template (murine polyomavirus) and the novel bat PyVs. An electrostatic potential difference pattern between the two bat VP1 pentamers was also revealed. Disaccharide molecular docking studies showed that the reference template and both bat PyVs possess the typical shallow sialic acid-binding site located between two VP1 subunits, with relevant oligosaccharide-binding affinities. The characterisation of these novel bat PyVs and the reported properties of their capsid proteins will potentially contribute in the elucidation of the conditions creating the host-pathogen restrictions associated with these viruses.
Assuntos
Quirópteros , Polyomavirus , Animais , Camundongos , Filogenia , Simulação de Acoplamento Molecular , Genoma Viral , Polyomavirus/genética , Polyomaviridae/genéticaRESUMO
Bovine adenoviruses (BAdV) are known to cause respiratory and/or intestinal disease in calves. Infection can manifest as acute outbreaks, but more often only sporadic cases occur. Here we describe the PCR detection and partial sequence characterization of several BAdVs found in sick or dead calves on different farms in Western Hungary. Intermittent diarrhoeal illnesses occurred after weaning among calves on several farms located up to 40 km apart. A high-sensitivity, broad-spectrum nested PCR, developed for the general detection of adenoviruses, gave positive results in four independent cases. Direct sequencing of PCR products showed clear results from only two samples, whereas sequences from the other two amplicons were mixed. Molecular cloning of these heterogeneous PCR products was performed to separate each DNA fragment therein. By sequencing several plasmid clones from both mixed samples, we were able to detect the simultaneous presence of two different BAdV types, namely types 6 and 10 classified into two separate (Atadenovirus and Mastadenovirus) genera. The sequence of one homogenous sample was identified as being derived also from BAdV-10, whereas the other sample contained a novel type, proposed to be BAdV-11. We demonstrated, for the very first time, the occurrence of the two latter virus types in continental Europe. Their appearance in Hungary marks a significant shift in the types of BAdVs actually circulating in the country. Considering the similarity of the pathological findings to those, attributed to BAdV-10 infections reported to date, the causative role of the viruses in these cases seems to be plausible. Phylogeny reconstruction further confirmed that BAdVs represent multiple genetic lineages.
Assuntos
Adenoviridae , Mastadenovirus , Bovinos , Animais , Sequência de Bases , Adenoviridae/genética , Mastadenovirus/genética , Reação em Cadeia da Polimerase/métodos , Reação em Cadeia da Polimerase/veterinária , Europa (Continente)/epidemiologiaRESUMO
A novel papillomavirus (PV) was detected in farmed wels catfish (Silurus glanis) in Hungary showing clinical signs resembling those of wels catfish herpesvirus disease. The whole genome of Silurus glanis papillomavirus 1 (SgPV1) was identified using next-generation sequencing. The 5,612-bp complete genome contains four predicted protein coding regions (E1, E2, L1, and L2), which seem to have homologues in every PV genome sequenced to date. Five complete fish PV genome sequences are available in the GenBank database. Their genomes range between 5,748 and 6,086 bp and contain the minimal PV backbone genes E1, E2, L2, and L1, unlike PVs of higher vertebrates, which have larger genomes (6.8-8.6 kbp) and additional (onco)genes. Considering the current species demarcation criteria for the family Papillomaviridae, the establishment of a novel species named "Nunpapillomavirus siluri" is proposed for the SgPV1 in a novel genus, "Nunpapillomavirus", in the subfamily Secondpapillomavirinae.
Assuntos
Peixes-Gato/virologia , Doenças dos Peixes/virologia , Papillomaviridae/classificação , Papillomaviridae/genética , Papillomaviridae/isolamento & purificação , Infecções por Papillomavirus/veterinária , Infecções por Papillomavirus/virologia , Sequenciamento Completo do Genoma , Animais , Genoma Viral , Sequenciamento de Nucleotídeos em Larga Escala , Hungria , Fases de Leitura Aberta , Filogenia , Pele/virologiaRESUMO
The aim of our study was to evaluate the possible effects of whole-body 1800 MHz GSM-like microwave exposure on male reproduction. After repeated exposure of mice to microwaves at 0.018-0.023 W/kg whole-body specific energy absorption rate (SAR) an elevated serum testosterone level was measured, but no microwave exposure related histopathological alteration could be detected in the reproductive organs. The in vitro steroidogenic response of 48 h Leydig cell cultures obtained from exposed animals did not differ from the controls, suggesting that Leydig cells were not the primary targets of the applied microwave exposure or direct action of microwaves on Leydig cells was temporary only. In exposed animals the red blood cell count and volume of packed red cells were also increased. Further investigations are required to clarify the mechanism of action of the applied microwave exposure on male mice, as well as to establish the biological significance of the observed phenomena.
Assuntos
Micro-Ondas/efeitos adversos , Hormônios Testiculares/biossíntese , Testículo/efeitos da radiação , Irradiação Corporal Total/métodos , Alanina Transaminase/sangue , Fosfatase Alcalina/sangue , Animais , Aspartato Aminotransferases/sangue , Peso Corporal/efeitos da radiação , Células Cultivadas , Colesterol/sangue , Contagem de Eritrócitos , Técnicas In Vitro , Células Intersticiais do Testículo/citologia , Células Intersticiais do Testículo/metabolismo , Células Intersticiais do Testículo/efeitos da radiação , Masculino , Camundongos , Camundongos Endogâmicos , Doses de Radiação , Testículo/metabolismo , Testosterona/biossíntese , Triglicerídeos/sangueRESUMO
The aim of this study was to evaluate the possible effects of in vivo 1800 MHz GSM-like exposure on male reproduction. In five separate experiments, male NMRI mice (35-41 g) were exposed (11-12 mice each) to 1800 MHz GSM-like radiation. The average power density was 100 microW/cm2, the estimated SAR was 0.018-0.023 W/kg. The animals were exposed ten times (over two weeks on workdays) and the duration of exposure was 2 h/day. On the day of the last treatment, mice were anesthetized with i.p. pentobarbital, and blood samples were taken for hematology, serum chemistry and serum testosterone (T) determinations (ELISA). Testicles, epididymes, adrenals, prostates and pituitary glands were removed for histology. One testicle of each animal was used for culture of Leydig cells. The cells were cultured for 48 h in the presence or absence of human chorionic gonadotropin (hCG) to evaluate the in vitro steroidogenic response of Leydig cells. In the exposed animals red blood cell count (RBC: 8.59+/-0.10 T/l, n=37) and volume of packed red cells (VPRC: 42.29+/-0.43%, n=37) were significantly higher (p<0.01) compared with the controls (RBC: 8.12+/-0.08 T/l, n=36; VPRC: 39.76+/-0.36%, n=36). The serum testosterone level of the exposed animals (7.85+/-1.08 ng/ml, n=56) was also significantly elevated (p<0.05) compared to the controls (5.12+/-0.79, n=52), while the in vitro steroidogenic capacity of the Leydig cells was unaltered. No significant differences in the other investigated variables were found between controls and exposed mice. Our results indicate that the applied GSM-like microwave exposure may induce slight, but statistically significant alterations in some hematological and endocrine parameters of male mice within the physiological range. Further investigations are required to establish the biological significance of these phenomena.
Assuntos
Biomarcadores/sangue , Campos Eletromagnéticos/efeitos adversos , Gônadas/efeitos da radiação , Micro-Ondas/efeitos adversos , Alanina Transaminase/sangue , Animais , Aspartato Aminotransferases/sangue , Colesterol/sangue , Relação Dose-Resposta à Radiação , Contagem de Eritrócitos , Hematócrito , Hemoglobinas/metabolismo , Contagem de Leucócitos , Masculino , Camundongos , Contagem de Plaquetas , Doses de Radiação , Triglicerídeos/sangueRESUMO
The main goal of this study was to evaluate the possible effect of whole-body magnetic field (MF) exposure on the steroidogenic capacity of Leydig cells in vitro. In four separate experiments, male CFLP mice were exposed to sinusoidal 50-Hz, 100-microT MF. The duration of exposure was 23.5 h/day over a period of 14 days. At the end of the exposure, interstitial (Leydig) cells were isolated from the testicles of the sham-exposed and exposed animals. The cells were cultured for 48 h in the presence or absence of 1, 10, or 100 mIU/ml human chorionic gonadotropin (hCG). The luteinizing hormone (LH) analog hCG was used to check the testosterone (T) response of the sham-exposed controls and to evaluate the possible effect of the whole-body MF exposure on the steroidogenic capacity of Leydig cells in vitro. Testosterone content of the culture media and blood sera was measured by radioimmunoassay (RIA). In the cultures obtained from MF-exposed animals, the hCG-stimulated T response was significantly higher (p < 0.01) compared with the sham-exposed controls, while the basal T production of cells and the level of serum T remained unaltered. No MF exposure-related histopathological alterations were found in testicles, epididymes, adrenals, prostates, and pituitary glands. The MF exposure did not affect the animal growth rate and the observed hematologic and serum chemical variables. Our results indicate a presumably direct effect of whole-body MF exposure on the hCG-stimulated steroidogenic response of mouse Leydig cells.
Assuntos
Campos Eletromagnéticos/efeitos adversos , Células Intersticiais do Testículo/efeitos da radiação , Irradiação Corporal Total/efeitos adversos , Glândulas Suprarrenais/fisiologia , Glândulas Suprarrenais/efeitos da radiação , Animais , Peso Corporal/efeitos da radiação , Células Cultivadas , Epididimo/patologia , Epididimo/efeitos da radiação , Humanos , Células Intersticiais do Testículo/química , Células Intersticiais do Testículo/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos , Hipófise/patologia , Hipófise/efeitos da radiação , Próstata/patologia , Próstata/efeitos da radiação , Doses de Radiação , Lesões Experimentais por Radiação/sangue , Radiação não Ionizante/efeitos adversos , Testículo/patologia , Testículo/efeitos da radiação , Testosterona/biossíntese , Testosterona/sangue , Irradiação Corporal Total/métodosRESUMO
The paper deals with the regulation of pesticides in agriculture, their EU marketing. National authorizations of pesticides are based on the Council Directive 91/414/EEC within the EU Member States. The authors outline the basic rules on the conditions and procedure for the authorization. They deal with the evaluation of toxicological data in particular.