ABSTRACT
BACKGROUND: The UK High-Consequence Infectious Diseases (HCID) Network of high-level isolation units provides care for patients with contact- or airborne-transmissible highly infectious and highly dangerous diseases. In most HCID units, the healthcare workers (HCWs) wear personal protective equipment (PPE) ensembles incorporating a powered air-purifying respirator (PAPR) for head and respiratory protection. Some PAPRs have components worn outside/over other PPE, necessitating decontamination of re-usable elements. Two alternative PAPRs, with all re-usable elements worn under PPE, were trialled in this study. AIM: To undertake scenario-based testing of PAPRs and PPE to determine usability, comfort and ability to remove contaminated PPE without personal cross-contamination. METHODS: Trained healthcare volunteers (N=20) wearing PAPR/PPE ensembles were sprayed with ultraviolet fluorescent markers. They undertook exercises to mimic patient care, and subsequently, after doffing the contaminated PPE following an established protocol, any personal cross-contamination was visualized under ultraviolet light. Participants also completed a questionnaire to gauge how comfortable they found the PPE. FINDINGS AND CONCLUSIONS: The ensembles were tested under extreme 'worst case scenario' conditions, augmented by physical and manual dexterity tests. Participating volunteers considered the exercise to be beneficial in terms of training and PPE evaluation. Data obtained, including feedback from questionnaires and doffing buddy observations, supported evidence-based decisions on the PAPR/PPE ensemble to be adopted by the HCID Network. One cross-contamination event was recorded in the ensemble chosen; this could be attributed to doffing error, and could therefore be eliminated with further practice.
Subject(s)
Communicable Diseases , Respiratory Protective Devices , Humans , Personal Protective Equipment , Health Personnel , Coloring AgentsABSTRACT
Ventilation is frequently used as a means for preventing the build up of flammable or toxic gases in enclosed spaces. The effectiveness of the ventilation often has to be considered as part of a safety case or risk assessment. In this paper methods for assessing ventilation effectiveness for hazardous area classification are examined. The analysis uses data produced from Computational Fluid Dynamics (CFD) simulations of low-pressure jet releases of flammable gas in a ventilated enclosure. The CFD model is validated against experimental measurements of gas releases in a ventilation-controlled test chamber. Good agreement is found between the model predictions and the experimental data. Analysis of the CFD results shows that the flammable gas cloud volume resulting from a leak is largely dependent on the mass release rate of flammable gas and the ventilation rate of the enclosure. The effectiveness of the ventilation for preventing the build up of flammable gas can therefore be assessed by considering the average gas concentration at the enclosure outlet(s). It is found that the ventilation rate of the enclosure provides a more useful measure of ventilation effectiveness than considering the enclosure air change rate.
Subject(s)
Gases , Ventilation , Models, MolecularABSTRACT
PURPOSE: If radiation-induced genetic instability is causal in mouse radiation leukaemogenesis, then genetic instability should be detectable in the irradiated target untransformed haemopoietic stem cell, and evidence of genetic instability detected in the clonal radiation-induced leukaemia. We have tested this hypothesis using the CBA/H mouse model of radiation-induced acute myeloid leukaemia (r-AML). MATERIALS AND METHODS: Fluorescence in situ hybridisation (FISH) was employed to screen for chromosomal aberrations in mouse 3 Gy X-ray-induced r-AMLs and in the clonal descendents of control and 3 Gy X-irradiated bone marrow haemopoietic stem cells using the in vitro clonogenic CFU-A colony assay. RESULTS: High levels of clonal non-specific chromosomal aberrations were detected in the r-AML (approximately 4-5 aberrations/r-AML), and ongoing chromosomal instability as defined by subclonal variants detected in 5/10 r-AML. A similar analysis of CFU-A colonies revealed chromosomal aberrations in 25% of colonies derived from irradiated bone marrow (2% in controls). However, 66% of the aberrant colonies (2% in controls) exhibited ongoing genetic instability as defined by non-clonal chromosomal aberrations. Overall, 6% (121/1884) of the CFU-A cells derived from irradiated bone marrow were aberrant (0.05% in controls) of which 12% (15/121) were subclonal. No one CFU-A cell exhibited aberrations on more than one of the three chromosomes painted. CONCLUSIONS: The high levels of non-specific genetic damage observed in the r-AMLs is therefore attributed to the accumulation of genetic lesions in the target haemopoietic stem cell over a longer time-scale after exposure than assessed in the in vitro CFU-A clonogenic assay. This is consistent with the long latency of the multi-stage radiation leukaemogenic process, and a role for radiation-induced genetic instability is inferred.
Subject(s)
Chromosome Aberrations , Chromosome Disorders , Hematopoietic Stem Cells/radiation effects , Leukemia, Myeloid, Acute/genetics , Leukemia, Radiation-Induced/genetics , Animals , Hematopoietic Stem Cells/ultrastructure , In Situ Hybridization, Fluorescence , Mice , Mice, Inbred CBA , X-RaysABSTRACT
PURPOSE: To study stable and unstable chromosomal aberrations in the haemopoietic cells of CBA/H mice after exposure to both high- and low-LET radiations. MATERIALS AND METHODS: Chromosomal aberrations were scored in the clonal progeny of X-, alpha- or non-irradiated short-term repopulating stem cells using the spleen colony-forming unit (CFU-S) assay, 12 days post-transplantation and in the bone marrow reconstituted by X-, neutron- or non-irradiated exogenous (transplanted) or endogenous (X- or neutron whole-body-irradiated) long-term repopulating stem cells for up to 24 months. RESULTS: Chromosomal instability was demonstrated in 3-6% of cells in all cases. After transplantation of X- or neutron-irradiated bone marrow approximately 8% of cells with stable aberrations were recorded at all times. After 3Gy X- or 0.5 Gy neutron- whole-body irradiation stable aberrations were detected in approximately 17 and 5% of cells respectively. CONCLUSIONS: Chromosomal instability induced in vitro can be transmitted in vivo by transplantation of haemopoietic stem cells exposed to high- or low-LET radiations. Comparable instability can be induced and shown to persist for the remaining lifetime after whole-body irradiation. There was no direct relationship between the expression of stable and unstable aberrations and significant interanimal variation in the expression of both stable and unstable aberrations.
Subject(s)
Chromosome Aberrations , Hematopoietic Stem Cells/radiation effects , Animals , Hematopoietic Stem Cell Transplantation , Hematopoietic Stem Cells/ultrastructure , Linear Energy Transfer , Mice , Mice, Inbred CBA , Whole-Body Irradiation , X-RaysABSTRACT
A robust method for obtaining chromosome preparations from individual haemopoietic colonies in semi-solid media is described. The accumulation of metaphases and the hypotonic treatment of cells were carried out in the culture dish and individual colonies were transferred onto poly-L-lysine-treated slides and fixed stepwise prior to staining. The technique produced high yields of well-spread metaphases facilitating clonal cytogenetic analysis.
Subject(s)
Chromosomes , Cytogenetics/methods , Hematopoietic Stem Cells , Animals , MiceABSTRACT
Moment-to-moment fluctuations of nucleus accumbens dopamine (DA) were determined in rats self-administering or passively receiving "yoked" intravenous infusions of D-amphetamine. The initial lever presses of each session caused elevations in DA concentration, usually to an initial peak that was not maintained throughout the rest of the session. As the initial ("loading") injections were metabolized, DA levels dropped toward baseline but were sustained at elevated plateaus by subsequent lever pressing that was spaced throughout the remainder of the 3 hr sessions. During this period, DA levels fluctuated phasically, time-locked to the cycle of periodic lever pressing. Consistent with the known pharmacological actions and dynamics of amphetamine, peak DA elevations were seen approximately 10-15 min after each injection, and the mean DA level was at a low point in the phasic cycle at the time of each new lever press. During extinction periods when saline was substituted for amphetamine, DA levels dropped steadily toward baseline levels despite a dramatic increase in (now-unrewarded) lever pressing. Noncontingent injections during extinction reinstated lever-pressing behavior and increased nucleus accumbens DA concentrations. These data are consistent with the hypothesis that under the conditions of this experiment-during periods of amphetamine intoxication in well-trained animals-the timing of amphetamine self-administration comes primarily under the control of extracellular DA concentrations. The probability of lever pressing during the maintenance phase is highest when DA concentrations fall near a characteristic trigger point, a trigger point that is significantly elevated above baseline, and falls as DA concentrations fall below or increase above that trigger point.
Subject(s)
Central Nervous System Stimulants/pharmacology , Dextroamphetamine/pharmacology , Dopamine/metabolism , Extinction, Psychological/drug effects , Nucleus Accumbens/drug effects , Reward , Animals , Catheterization, Central Venous , Male , Microdialysis , Nucleus Accumbens/metabolism , Rats , Rats, Long-Evans , Self AdministrationABSTRACT
We have demonstrated chromosomal instability in the clonal descendants of hemopoietic stem cells after irradiating murine bone marrow with alpha-particles. However, because cells that are irradiated by alpha-particles are defined by a Poisson distribution of individual particle traversals, there is an inevitable proportion of unirradiated cells in the surviving population. The calculated expected proportions of irradiated and nonirradiated cells indicate that the number of clonogenic cells transmitting chromosomal instability is greater than the number expected to be hit and survive. To investigate further this discrepancy, we studied the effects of interposing a grid between the cells and the alpha-particle source so that the surviving population consists predominantly of untraversed stem cells. Comparison with the same irradiation conditions without the grid reveals that the same level of instability is induced. The data confirm that alpha-particles induce chromosomal instability but instability is demonstrated in the progeny of nonirradiated stem cells and must be due to unexpected interactions between irradiated and nonirradiated cells. This untargeted effect has important implications for mechanistic studies of radiation action and for assessment of radiation risk.
Subject(s)
Abnormalities, Radiation-Induced/genetics , Alpha Particles/adverse effects , Chromosomes/radiation effects , Animals , Clone Cells/radiation effects , Dose-Response Relationship, Radiation , Hematopoietic Stem Cells/radiation effects , Male , MiceSubject(s)
Hospitalization , Tuberculosis/therapy , Adult , Cost of Illness , Female , Humans , Malawi , MaleABSTRACT
Fluctuations in extracellular dopamine and DOPAC levels in nucleus accumbens septi (NAS) were monitored in 1-min microdialysis samples taken from rats engaged in intravenous cocaine self-administration. For four rats the dose per injection was fixed at 2.0 mg/kg; for four others the dose per injection was varied irregularly, from one response to the next, between three levels (0.5, 1.0 and 2.0 mg/kg). Regardless of the dosing regimen, extracellular dopamine levels were tonically elevated by 200-800% within the cocaine self-administration periods, fluctuating phasically within this range between responses. In the fixed dose condition, the phasic increases following each injection (and the phasic decreases preceding them) averaged approximately 50% of the mean tonic elevation. Phasic fluctuations in dopamine levels remained time-locked to lever-presses even when response rate was irregular, because of the variable dose condition. In the variable dose condition greater increases in dopamine and longer inter-response times followed injections of the higher doses; dopamine fluctuations were consistent with the multiple-infusion pharmacokinetics of cocaine. DOPAC levels showed a slow tonic depression during cocaine self-administration, but individual injections were not associated with discernible phasic fluctuations of DOPAC. These data are consistent with the hypothesis that falling dopamine levels trigger successive responses in the intravenous cocaine self-administration paradigm, but inconsistent with the notion that extracellular dopamine levels are depleted at the times within sessions when the animal initiates drug-seeking responses.
Subject(s)
Cocaine/pharmacology , Dopamine/metabolism , Nucleus Accumbens/drug effects , 3,4-Dihydroxyphenylacetic Acid/metabolism , Animals , Dose-Response Relationship, Drug , Injections, Intravenous , Male , Microdialysis , Rats , Rats, Inbred Strains , Self Administration , Time FactorsABSTRACT
In vivo microdialysis was used to assess the involvement of ventral tegmental area (VTA) mu, delta, and kappa opioid receptors in modulation of basal extracellular ventral striatal dopamine (DA) and DA-metabolite concentrations. Independent groups of chloral hydrate-anesthetized rats were given VTA microinjections of selective opioid agonists, and extracellular ventral striatal DA and DA-metabolite concentrations were assayed using HPLC. VTA microinjections of [D-Ala2, N-Me-Phe4-Gly5-ol]-enkephalin (DAMGO; a mu agonist) and [D-Pen2, D-Pen5]-enkephalin (DDDPE; a delta agonist) each caused dose-orderly increases in ventral striatal DA and DA-metabolite concentrations. The effective concentrations of DPDPE were 100- to 1000-fold higher than the effective concentrations of DAMGO. VTA microinjections of (trans-(dl)-3,4-dichloro-N-methyl-N-[2-(1-pyrrolidinyl)cyclo-hexyl]- benzeneacetamide) methane sulfonate hydrate (U-50,488H); a kappa agonist) failed to alter ventral striatal DA concentrations at any dose tested, but subsequent systemic injections significantly decreased DA and DA-metabolite concentrations. Pretreatment with VTA microinjections of 17-cyclopropylmethyl-6,7-dehydro-4,5-epoxy-3,14-dihydroxy-6,7,2',3'- indolmorphinan hydrochloride (naltrindole; a delta antagonist) (delta antagonist) antagonized VTA DPDPE-mediated increases in ventral striatal DA and DA-metabolite concentrations but failed to antagonize VTA DAMGO-mediated increases. Pretreatment with D-Pen-Cys-Tyr-D-Trp-Orn-Thr-Pen-Thr-NH2 (CTOP; a mu antagonist) antagonized VTA DAMGO-mediated increases but failed to antagonize VTA DPDPE-mediated increases. Thus both mu and delta receptor agonist appear capable of increasing ventral striatal DA and DA-metabolite concentrations through selective actions on their preferred class of opioid receptors in the VTA. The increases in ventral striatal DA and DA-metabolite concentrations that are seen after systemic treatment with kappa opioid agonists appear not to involve VTA opioid receptors.
Subject(s)
Dopamine/metabolism , Limbic System/metabolism , Receptors, Opioid/physiology , Tegmentum Mesencephali/ultrastructure , 3,4-Dichloro-N-methyl-N-(2-(1-pyrrolidinyl)-cyclohexyl)-benzeneacetamide, (trans)-Isomer , 3,4-Dihydroxyphenylacetic Acid/metabolism , Amino Acid Sequence , Analgesics/pharmacology , Animals , Chromatography, High Pressure Liquid , Corpus Striatum/metabolism , Dialysis/methods , Dopamine/physiology , Enkephalin, Ala(2)-MePhe(4)-Gly(5)- , Enkephalin, D-Penicillamine (2,5)- , Enkephalins/pharmacology , Homovanillic Acid/metabolism , Hydroxyindoleacetic Acid/metabolism , Limbic System/drug effects , Male , Microinjections , Molecular Sequence Data , Naltrexone/analogs & derivatives , Naltrexone/pharmacology , Narcotic Antagonists , Pyrrolidines/pharmacology , Rats , Rats, Inbred Strains , Receptors, Opioid/drug effects , Tegmentum Mesencephali/drug effects , Tegmentum Mesencephali/physiologyABSTRACT
Transmissible gastroenteritis (TGE) infection causes 65% of infectious piglet diarrhoea in Taiwan. A virulent Taiwanese strain, TFI, of transmissible gastroenteritis virus (TGEV) from a field outbreak was isolated in cell culture and plaque purified. Phenotypic differences were observed in the ability of TFI to infect certain cell lines. TGEV strains TLM-83 (PRCV Belgium), TO-163 (TGEV Japan) and Purdue-115 (TGEV USA) infected both ST (swine testis) and RPTG (pig kidney) cell lines whereas TFI infected ST but not RPTG cells. To investigate this phenotypic variation cDNA was generated from TFI genomic and amplified by PCR with oligonucleotides derived from published TGEV sequence data. An 8.4kb cDNA derived from the 3'-end of the TFI genome was sequenced. Eight ORFs, corresponding to the three structural protein genes, four potential genes and the 3'-end of an incomplete ORF whose amino acid sequence corresponded to the carboxyl end of the 1b subunit of the polymerase gene, were identified on the TFI sequence. The overall sequence similarity of TFI with the other TGEV strains was over 97%. However, several deletions, insertions and point mutations were found on the TFI sequence when compared with other TGEV strains. The TFI S protein was found to contain 1449 amino acids, as also identified for the FS772/70 and Miller TGEV strains, but two amino acids longer than the Purdue S protein. The TFI ORF-3a gene encodes 72 amino acids, however, a 37 nucleotide deletion was found 16 nucleotides downstream of the TFI ORF-3a stop codon.(ABSTRACT TRUNCATED AT 250 WORDS)
Subject(s)
Genome, Viral , Transmissible gastroenteritis virus/genetics , Amino Acid Sequence , Base Sequence , Molecular Sequence Data , Open Reading Frames , Polymerase Chain Reaction , Taiwan , Transmissible gastroenteritis virus/pathogenicity , VirulenceABSTRACT
Previous studies on different transmissible gastroenteritis virus (TGEV) strains, including porcine respiratory coronavirus (PRCV), have identified regions within the genome that are polymorphic as regards insertions and deletions. For example the 672 base deletion within the S gene and multiple deletions 5', within and 3' of the ORF-3a gene were detected in strains of PRCV. The presence of deletions may be associated with a change in the virulence, attenuation or tissue tropism of the isolate. The Nouzilly (188-SG) TGEV vaccine strain was attenuated by passage of a cell culture adapted virulent isolate D-52 188 times through swine testis cells after treatment with gastric juice. PCR amplification with oligonucleotides, corresponding to known TGEV sequences, were used to analyse D-52 and 188-SG for genetic variation. Results with several pairs of oligonucleotides within the first 1565 nucleotides of the S gene did not identify a deletion within this region of the genome from either strain. However, oligonucleotides directed against the ORF-3a/3b region detected a deletion of about 250 nucleotides within the 188-SG genome but not in the D-52 genome. Since all the attenuated TGEV strains so far sequenced, PRCV, Miller SP and 188-SG, contained deletions within the ORF-3a/3b, it would suggest that this region of the TGEV genome is involved in regulating viral virulence.
Subject(s)
Genome, Viral , Polymerase Chain Reaction , Transmissible gastroenteritis virus/classification , Amino Acid Sequence , Animals , Base Sequence , Cell Line , Molecular Sequence Data , Open Reading Frames , Phenotype , Sequence Alignment , Sequence Homology , Species Specificity , Swine , Transmissible gastroenteritis virus/geneticsABSTRACT
Coronavirus proteins are translated from a nested set of subgenomic mRNAs which have common 3' termini with unique 5' extensions. Evidence suggests that coronavirus mRNAs are generated by a mechanism of leader primed transcription. Leader RNA binds to consensus sequences upstream of each gene on full length negative strand viral RNA and transcription proceeds to the 5' end of the negative strand to produce the nested set of mRNAs. Even though this gives rise to polycistronic mRNA species only the 5' extension of each mRNA is translated to give the viral proteins. The leader RNA for TGEV is about 90 nucleotides long and contains the sequence which recognises the leader binding sites on the negative strand RNA. Evidence suggests that the length of the leader binding sequence may be involved in transcriptional control of individual mRNAs. In order to investigate this a virus specific mRNA isolation method was developed to measure the relative amounts of mRNAs synthesised during an infection of LLC-PK1 cells with TGEV (strain FS772/70). Thus the relative quantity of each mRNA can be determined and correlated with the variation in size of the leader binding site.
Subject(s)
Gene Expression Regulation, Viral , RNA, Messenger/genetics , RNA, Viral/genetics , Transcription, Genetic , Transmissible gastroenteritis virus/genetics , Animals , Base Sequence , Cell Line , Microspheres , Molecular Sequence Data , RNA, Messenger/biosynthesis , RNA, Viral/biosynthesis , SwineSubject(s)
Cat-Scratch Disease/diagnosis , Erythema Nodosum/etiology , Adult , Cat-Scratch Disease/complications , Female , HumansABSTRACT
Microdialysis and high-performance liquid chromatography with electrochemical detection were used to determine extracellular levels of dopamine following ventral tegmental morphine injections into chloral hydrate-anesthetized rats. Morphine (13.2 nanomoles in 0.5 microliter of Ringer's solution) caused 50-150% increases in nucleus accumbens dopamine and metabolites; latency for the effect was on the order of 15 min with peak effects occurring in 30-50 min. Contralateral dopamine levels were influenced only minimally. These data suggest opiate receptors in or near the ventral tegmental area as sites of the opioid action that is responsible for opioid activation of the mesolimbic dopamine system.
Subject(s)
Dopamine/metabolism , Morphine/pharmacology , Nucleus Accumbens/metabolism , Tegmentum Mesencephali , Animals , Chromatography, High Pressure Liquid , Dialysis , Electrochemistry , Injections , Male , Microchemistry , Morphine/administration & dosage , Nucleus Accumbens/drug effects , RatsABSTRACT
Two new monoclonal antibodies (CC17 and CC29) raised against bovine thymocytes are described. The antibodies, both of which were IgG1, recognize a molecule of approximately 67,000 molecular weight on bovine T cells. They react T cells in peripheral blood, the lymph node paracortex and the periateriolar lymphoid sheath in the spleen. Both the cortex and medulla of the thymus are stained but the medulla reacts more intensely. They do not stain B cells in peripheral blood, the ileal Peyer's patch, the cortex or the primary follicles in lymph nodes. No activity was found on cells outside the lymphoid system, i.e. monocytes, alveolar macrophages or endothelial and epithelial tissue. The antigen recognized is considered to be the bovine homologue of CD5 (T1) in humans and Lyt1 in mice. The mAbs appear to be particularly useful for detecting cells in the peripheral blood of young calves which are of the T cell lineage but do not express BoT2 or the mature pan T cell antigen recognized by mAb IL-A27 and may thus allow identification of a population of bovine lymphocytes previously described as null cells.
Subject(s)
Antibodies, Monoclonal/immunology , Antigens, Differentiation, T-Lymphocyte/immunology , Cattle/immunology , T-Lymphocytes/immunology , Animals , Antibodies, Monoclonal/biosynthesis , Antigens, Surface/immunology , Cell Line , Cell Separation , Cells, Cultured , Flow Cytometry , Fluorescent Antibody Technique , Hybridomas , Immunoenzyme Techniques , Immunohistochemistry , Mice , Molecular Weight , Precipitin TestsABSTRACT
Analysis of the genomic dsRNA of rotaviruses isolated from calves with subclinical infections has revealed eight calves excreting group A viruses with unusual genome profiles. Two of these virus strains, C7-176 and C7-183, were grown and cloned by plaque purification in cell culture. Examination of their genome profiles after cloning showed that the unusual pattern had been retained. They were without RNA segment 11 but had an extra band (6a) migrating between segments 6 and 7. However, they contained the triplet of segments 7, 8 and 9, of similar size, which is characteristic of group A rotaviruses. The number and mol. wt. of the intracellular polypeptides induced by these viruses were similar to those of the bovine group A rotavirus UK strain. Analysis of the RNA transcripts produced by the transcription in vitro of purified C7-183 virus showed that segment 6a produced a large RNA transcript of corresponding size. After isolation, this transcript was translated in a rabbit reticulocyte lysate preparation and yielded a single polypeptide, vp11, equivalent to the product of segment 11 of rotaviruses with typical genome profiles.
Subject(s)
Cattle Diseases/microbiology , Genes, Viral , Rotavirus Infections/veterinary , Rotavirus/isolation & purification , Animals , Cattle , RNA, Double-Stranded/genetics , Rotavirus/genetics , Viral Proteins/analysis , Viral Proteins/geneticsABSTRACT
An outline of the clinical diseases that arise following BVDV infection is given. Isolates of BVDV can be separated into two forms non-cytopathic and cytopathic depending on their effect on cell cultures. In utero infection of the foetus with non-cytopathic virus may result in a number of syndromes, such as abortions, stillbirths or weak calves. It may also result in the birth of calves with a persistent viraemia and these animals may later develop mucosal disease as a result of superinfection with a "homologous" cytopathic strain of BVDV. Acute infection of seronegative animals in usually mild and subclinical. A chronic disease also occurs and this can be protracted with progressive wastage and diarrhoea. This condition has not yet been well defined but it is suggested that it may be the result of superinfection of a viraemic animal with a "heterologous" cytopathic strain of virus. The maintenance of non-cytopathic virus within the cattle population can be either by the slow spread following acute infections of seronegative animals or, more importantly, by spread from persistently viraemic cattle. The cytopathic virus is usually found in association with cases of mucosal disease and may be maintained in the population only by continually arising, possibly by mutation, from the non-cytopathic virus. It is recommended that persistently viraemic animals are eliminated from the herd to avoid in utero infections and the possibility of subsequent mucosal disease.
Subject(s)
Bovine Virus Diarrhea-Mucosal Disease/etiology , Cattle Diseases/etiology , Animals , Bovine Virus Diarrhea-Mucosal Disease/epidemiology , Bovine Virus Diarrhea-Mucosal Disease/pathology , CattleABSTRACT
Rotaviruses isolated from 43 sub-clinically infected calves from a single farm were analysed by genome profile analysis. The isolates showed genomic variation and eight different profiles were observed, including one which was atypical for Group A rotaviruses. The 3' terminal labelling method for the analysis of genome profiles used in this study required only 1 ng of viral RNA, an increase of 1000-fold in sensitivity over ethidium bromide staining for detecting all rotavirus genome segments. However, dual infections involving two rotaviruses with distinct profiles could not be detected if the concentrations of the viruses differed by greater than 10-fold.