Electronic Library of Scientific Literature
Volume 42 / June 1998 / number 3
E.TUJULIN, A. MACELLARO, B. LILLIEHÖÖK, L. NORLANDER
Department of Microbiology,
Department of Biomedicine, Division of NBC Defence, Defence Research Establishment, S-901 82 Umea, Sweden;
Department of Pathology, Swedish University of Agricultural Sciences, P.O. Box 7028, S-750 07 Uppsala, Sweden
Summary. – The obligate intracellular rickettsia Coxiella
burnetii has previously been reported to reach the intravacuolar compartment
of host cells by phagocytosis. With the aim to further examine the mechanisms
of C. burnetii internalisation, macrophage monolayers were treated
with well characterised inhibitors of endocytosis. The treatment with two
general inhibitors, colchicine and methylamine, resulted in a pronounced
dose-dependent decrease of radiolabelled phase II rickettsiae retained
from the intracellular fraction. A third inhibitor used, amiloride, has
been reported to reduce effectively clathrin-independent pinocytic pathways.
The internalisation of C. burnetii was shown to be substantially
reduced also by amiloride and the effect was dependent on its concentration.
The passive role of C. burnetii in the internalisation was verified by using heat-killed C. burnetii. Host cells treated with either of the three inhibitors (amiloride, colchicine and methylamine) showed a similar reduction of intracellular C. burnetii after exposure to killed as well as live organisms.The data presented indicate that different endocytic mechanisms, pinocytosis as well as phagocytosis, may mediate the uptake of C. burnetii by a host cell.
Key words: Coxiella burnetii; internalisation; endocytosis
Acta virologica 42: 125 – 131, 1998
T.A. OWOLABI, M.A. TAIWO, G.A. THOTTAPPILLY, S.A. SHOYINKA, E. PROLL, F. RABENSTEIN
Department of Biological Sciences, University of Calabar, Calabar,
Department of Biological Sciences, University of Lagos, Akoka, Lagos, Nigeria;
International Institute of Tropical Agriculture, Ibadan, Nigeria;
Institute of Agricultural Research and Training, Obafemi Awolowo University, Moor Plantation, Ibadan;
Institute for Resistance Research and Pathogen Diagnostic, Aschersleben, Germany
Summary. – A sap transmissible virus, causing mosaic and leaf curl disease of Celosia argentea, was isolated at vegetable farms in Amuwo Odofin, Tejuoso, and Abule Ado, Lagos, Nigeria. The virus had a restricted host range confined to a few species of the Amaranthaceae, Chenopodiaceae and Solanaceae families. It failed to infect several other species of the Aizoaceae, Brassicaceae, Cucurbitaceae, Fabaceae, Lamiaceae, Malvaceae, Poaceae and Tiliaceae families. The virus was transmitted in a non-persistent manner by Aphis spiraecola and Toxoptera citricidus but not by eight other aphid species tested. There was no evidence of transmission by seeds of C. argentae varieties.The viral coat protein had a relative molecular mass (Mr) of about 30.2 K. Electron microscopy of purified virus preparations revealed flexuous rod shaped particles of about 750 nm in length. Serological studies were performed using the enzyme-linked immunosorbent assay (ELISA), immunosorbent electron microscopy (ISEM) and Western blot analysis. The virus reacted positively with an universal potyvirus group monoclonal antibody (MoAb) and MoAb P-3-3H8 raised against peanut stripe potyvirus. It also reacted with polyclonal antibodies raised against several potyviruses including asparagus virus-1 (AV-1), turnip mosaic virus (TuMV), maize dwarf mosaic virus (MDMV), watermelon mosaic virus (WMV-2), plum pox virus (PPV), soybean mosaic virus (SoyMV), lettuce mosaic virus (LMV), bean common mosaic virus (BCMV) and beet mosaic virus (BMV) in at least one of the serological assays used. On the basis of host range, mode of transmission, and available literature data, the celosia virus seems to be different from potyviruses previously reported to infect vegetables in Nigeria. The name celosia mosaic virus (ClMV) has been proposed for this virus.
Key words: celosia mosaic virus; potyvirus; celosia argentea;
host range; transmission; aphids; Nigeria
Acta virologica 42: 133 – 139, 1998
A. KHANNA, C.D. PODURI, P. MURUGAN, S. KUMAR, V.S. SUGUNAN, K.T. SHENOY, M.R. DAS
Rajiv Gandhi Centre for Biotechnology, Jagathy, Trivandrum 695 014,
Department of Gasteroenterology, Medical College Hospital, Trivandrum 695 011, Kerala, India
Summary. – A peptide-based enzyme immunoassay (PBEIA) has been developed using synthetic peptides whose sequences were selected from the core, envelope and non-structural regions of the prototype hepatitis C virus (HCV) genome. Results of PBEIA of sera obtained from several patients with various liver disorders were compared to those of commercial enzyme-linked immunosorbent assays (ELISA) and reverse transcription-polymerase chain reaction (RT-PCR). A large number of samples, which were repeatedly negative in commercial ELISA, were positive in PBEIA. There was a good correlation between the results of PBEIA and RT-PCR. The developed PBEIA proved to be a sensitive assay that had high specificity and was capable of detecting antibodies in various HCV-related liver disorders including the acute phase of the infection.
Key words: hepatitis C virus; epitopes; antibodies; peptide-based
immunoassay; synthetic peptides; ELISA; RT-PCR
Acta virologica 42: 141 – 145, 1998
J.J. ELLIOTT, D.L. RUBLE, G.M. ZAUCHA, G.P. JAAX, D.M. WAAG
Veterinary Medicine Division,
Bacteriology Division, United States Army Medical Research Institute of Infectious Diseases (USAMRIID), Fort Detrick, Frederick, MD, USA
Summary. – Coxiella burnetii phase I whole cell vaccine (WCV) is associated with risk of severe local delayed-type hypersensitivity (DTH) reactions in previously immunized individuals or those sensitized by natural exposure. We compared this vaccine to another investigational vaccine derived by chloroform-methanol extraction of phase I whole cells (chloroform-methanol residue vaccine, CMRV). Hairless guinea pigs, sensitized with either WCV or CMRV, were given 60, 600 and 6,000 ng of WCV or CMRV in an intradermal (i.d.) skin test. The i.d. administration of WCV consistently caused more host reactions than comparable doses of CMRV in guinea pigs sensitized with either WCW or CMRV, suggesting that CMRV may be a safer vaccine. However, the CMRV was not innocuous and caused significant indurated lesions and micro-abscesses at the 600 ng and 6,000 ng skin test sites.
Key words: Q fever; Coxiella burnetii; chloroform-methanol residue
vaccine; whole cell vaccine; skin test
Acta virologica 42: 147 – 155, 1998
M.M. EL-SAGEYER, A. SZENDRÖI, E. HÜTTER, M. ÚJ, G. SZÜCS, I. MEZEY, I. TÓTH, A. KÁTAI, Z. KAPILLER, G. PÁLL, G. PETRÁS, E. SZALAY, I. MIHÁLY, S. GOUROVA, G. BERENCSI
Department of Biology, Faculty of Science, University of Garyounis,
Benghazi, Libyan Jamahiriya;
Department of Virology, and Department of Hospital Hygiene, B. Johan National Centre for Epidemiology, Gyáli St. 2-6, 1966 Budapest, Hungary;
Department of Virology, Regional State Public Health Service, Pécs,Hungary;
Regional State Public Health Service, Debrecen, Hungary;
Regional State Public Health Service, Szeged, Hungary;
St. László Hospital of Infectious Diseases, Budapest, Hungary;
National Centre for Infectious and Parasitic Diseases, Sofia, Bulgaria
Summary. – Echovirus 11' (prime) isolates from an epidemic of haemorrhagic syndrome in departments of obstetrics in Hungary have been characterised. The leading component of the clinical disease was carditis and its lethal outcome occurred in 13 newborn babies. Maternal immunity was found to be absent even in women of 41 years of age. The application of monovalent oral poliovirus type 1 vaccine prevented the progress of the epidemic within two weeks. Nevertheless, a serological survey among primovacinees of 3 – 15 months of age revealed that 20% of the babies seroconverted without clinical symptoms during the epidemic. Serological evidence showed that the echovirus 11' infection was unable to interfere with the efficacy of oral poliovirus vaccine (OPV), since seroconversion rates of primovaccinees did not differ significantly from those in the group seroconverted also to echovirus 11' during the vaccination campaign. A 440 nucleotide (nt) fragment of the 5'-non-translated region of 12 epidemic echovirus 11' isolates and 26 echovirus prototype strains was amplified by a nested reverse transcription-polymerase chain reaction (RT-PCR) and analysed using three different restriction endonucleases. The 5'-regions of the echovirus 11' isolates were found to be identical to each other but different from that of the prototype echovirus 11 (Gregory) strain. The results indicate that echovirus 11' isolates underwent genetic changes in the 5'-end and P1 region of the genome before the onset of the epidemic.
Key words: echovirus 11'; haemorhagic syndrome; newborn habies;
Acta virologica 42: 157 – 166, 1998
A.K. KUNDU, K. OHSHIMA, N. SAKO
Laboratory of Plant Virology, Faculty of Agriculture, Saga University, 1-banchi, Honjo-machi, Saga 840-8502, Japan
Summary. – Serological differences between two zucchini yellow mosaic virus (ZYMV) isolates (ZYMV-169 and ZYMV-M) obtained from two distinct geographical locations in Japan were determined by mapping epitopes on the coat proteins (CPs) of the two isolates. A total of 45 monoclonal antibodies (MAbs) against the two isolates were produced and the epitopes on the CPs were delineated by reacting these MAbs with trypsin-treated ZYMV particles and Escherichia coli-expressed ZYMV CP fragments. Six MAbs of groups I-a and I-b, specific for ZYMV-169, recognised two epitopes in the N-terminal region at amino acids (aa) 1–28 and 6–41 of ZYMV-169 CP. Fourteen MAbs of group II, specific for ZYMV-M, recognised epitopes in the N-terminal region of ZYMV-M CP. Twenty-one MAbs of groups III-a, III-b(i), III-b(ii), and III-b(iii), reacting with both isolates, recognised four epitopes; one epitope was located in the N-terminal region at aa 6–28 and the remaining three epitopes were located in the core region at aa 42–95, 171–227 and 228–259 of ZYMV CPs.
Key words: zucchini yellow mosaic virus; coat protein; monoclonal
antibody; fusion protein; epitope
Acta virologica 42: 167 – 173, 1998
J. BLAHOVÁ, K. KRÁLIKOVÁ, V. KRČMÉRY, SR., A. MIKOVIČOVÁ, N. BARTONÍKOVÁ
Institute of Preventive and Clinical Medicine, Limbová 14, 833 01
Bratislava, Slovak Republic;
Regional Hospital, Nové Zámky, Slovak Republic;
Bata's Hospital, Zlín, Czech Republic
Summary. – Two high frequency transduction (HFT) phage isolates, obtained from seriously ill patients, transducing individual determinants of antibiotic resistance with a frequency of 10-5 (phage isolate AP-103) and 10-6 (phage isolate AP-343), are described. The frequency of transduction depended on the transduced determinant(s) of resistance used for the detection of transductants and on the individual recipient antibiotic-susceptible strain of Pseudomonas aeruginosa (PAO and/or ML series). A multiple-antibiotic resistance was transduced by the phage isolate AP-343 to all tested recipient strains. The appearence of such phages in clinical conditions with an unusually high frequency of transduction might contribute to the dissemination of antibiotic resistance genes among nosocomial strains of P. aeruginosa. The existence of HFT phages might reflect an increased efficiency of transduction of antibiotic resistance among P. aeruginosa strains, and thus an increased risk of spread of antibiotic resistance even to recently introduced anti-pseudomonadal antibiotics among pseudomonads with unfavourable and unwanted epidemiological consequences in hospital conditions.
Key words: Pseudomonas aeruginosa; antibiotic resistance; transduction
Acta virologica 42: 175 – 179, 1998
M.A. QUEVEDO DIAZ, M. LUKÁČOVÁ
Institute of Virology, Dúbravská cesta 9, 842 46 Bratislava, Slovak Republic
Summary. – The influence of the number of passages in chick embryo yolk sac (EPs) on the properties of the lipopolysaccharide (LPS) and other antigens of Coxiella burnetii Priscilla strain in polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulphate (SDS-PAGE), immunoblot analysis, enzyme-linked immunosorbent assay (ELISA) and complement-fixation reaction (CFR) test has been studied. Three phases in the phase variation of Coxiella burnetii could be distinguished by these methods: phase I lasting up to the 20th passage (EP 20), intermediate phase corresponding to EP 20-EP 70, and phase II beginning at EP 80. The changes in LPS were more marked than those in proteins which conserved their immunoblot profile up to EP 80. The phase II was clearly demonstrated by all the methods used.
Key words: Coxiella burnetii; phase variation; lipopolysaccharide;
Acta virologica 42: 181 – 185, 1998
Departamento de Virologia, Instituto Oswaldo Cruz/FIOCRUZ, Av. Brazil 4365, 21040-360 Rio de Janeiro, RJ, Brazil
Summary. – The oral poliovirus vaccine (OPV) has been effectively used in the reduction and control of poliomyelitis cases on the planet. Despite several advantages of using the attenuated OPV strains, the rare occurrence of vaccine-associated paralytic poliomyelitis (VAPP) cases in vaccine recipients and their susceptible contacts is a disadvantage. Molecular biology studies of polioviruses isolated from stool and central nervous systém (CNS) of patients with VAPP have confirmed the vaccine origin of the isolates and demonstrated genomic modifications known or suspected to increase the neurovirulence. Similar genomic modifications have also been identified in OPV-derived strains isolated from healthy vaccinees and healthy contacts, suggesting that host factors are also involved in the establishment of poliomyelitis. Other neurologic complications such as meningitis, encephalitis, convulsions, transverse myelitis and Guillain-Barré syndrome have also been rarely associated with the use of this vaccine. The characterization of polioviruses isolated from such cases has demonstrated their OPV origin.
Key words: oral poliovirus vaccine; vaccine-associated cases;
Acta virologica 42: 187 – 195, 1998
D. KUBELKOVÁ, J. ŠPAK
Department of Plant Virology, Institute of Plant Molecular Biology, Academy of Sciences of the Czech Republic, Branišovská 31, 370 05 České Budějovice, Czech Republic
Key words: raspberry bushy dwarf virus; seed transmission; ELISA
Acta virologica 42: 195 – 196, 1998