RÉSUMÉ
The UK military medical treatment facility (MTF) that deployed to the United Nations Mission in South Sudan in 2017 was based on a facility that can provide damage control surgery and resuscitation for soldiers with ballistic trauma injuries. It had to be supplemented with additional medical equipment and drugs that could support a peacekeeping mission in Africa. The clinicians used this experience and opportunity to review the critical care capability of UK Army Medical Services forward MTFs and recommend changes to reflect the increasing contemporaneous need on recent deployments to support more casualties with medical, infectious diseases and other non-battle injuries and illnesses. A concurrent review of the facility's critical care transfer equipment was also undertaken and allowed it to be adapted for use as either transfer equipment or as a critical care surge capability, to increase the facility's critical care capacity.
Sujet(s)
Médecine militaire , Personnel militaire , Soins de réanimation , Humains , Réanimation , Soudan du SudRÉSUMÉ
The location of care for many brain-injured patients has changed since 2012 following the development of major trauma centres. Advances in management of ischaemic stroke have led to the urgent transfer of many more patients. The basis of care has remained largely unchanged, however, with emphasis on maintaining adequate cerebral perfusion as the key to preventing secondary injury. Organisational aspects and training for transfers are highlighted, and we have included an expanded section on paediatric transfers. We have also provided a table with suggested blood pressure parameters for the common types of brain injury but acknowledge that there is little evidence for many of our recommendations. These guidelines remain a mix of evidence-based and consensus-based statements. We have received assistance from many organisations representing clinicians who care for these patients, and we believe our views represent the best of current thinking and opinion. We encourage departments to review their own practice using our suggestions for audit and quality improvement.
Sujet(s)
Lésions encéphaliques/thérapie , Transfert de patient/méthodes , Accident vasculaire cérébral/thérapie , Transport sanitaire/méthodes , Anesthésiologie , Anesthésistes , Soins de réanimation , Humains , Sociétés médicalesRÉSUMÉ
BACKGROUND: Crohn's disease (CD) and ulcerative colitis (UC) are two pathotypes of inflammatory bowel disease (IBD) with unique pathology, risk factors and significant morbidity. AIM: To estimate incidence and identify IBD risk factors in a US military population, a healthy subset of the US population, using information from the Millennium Cohort Study. METHODS: Incident IBD was identified from medical encounters from 2001 to 2009 or by self-report. Our primary risk factor of interest, infectious gastroenteritis, was identified from medical encounters and self-reported post-deployment health assessments. Other potential risk factors were assessed using self-reported survey responses and military personnel files. Hazard ratios were estimated using Cox proportional hazards analysis. RESULTS: We estimated 23.2 and 21.9 diagnoses per 100 000 person-years, respectively, for CD and UC. For CD, significant risk factors included [adjusted hazard ratio (aHR), 95% confidence interval]: current smoking (aHR: 2.7, 1.4-5.1), two life stressors (aHR: 2.8, 1.4-5.6) and prior irritable bowel syndrome (aHR: 4.7, 1.5-15.2). There was no significant association with prior infectious gastroenteritis. There was an apparent dose-response relationship between UC risk and an increasing number of life stressors. In addition, antecedent infectious gastroenteritis was associated with almost a three-fold increase in UC risk (aHR: 2.9, 1.4-6.0). Moderate alcohol consumption (aHR: 0.4, 0.2-0.6) was associated with lower UC risk. CONCLUSIONS: Stressful conditions and the high risk of infectious gastroenteritis in deployment operations may play a role in the development of IBD in military populations. However, observed differences in risk factors for UC and CD warrant further investigation.
Sujet(s)
Gastroentérite/épidémiologie , Infections/épidémiologie , Maladies inflammatoires intestinales/épidémiologie , Personnel militaire/statistiques et données numériques , Adulte , Sujet âgé , Études de cohortes , Rectocolite hémorragique/complications , Rectocolite hémorragique/épidémiologie , Maladie de Crohn/complications , Maladie de Crohn/épidémiologie , Femelle , Gastroentérite/complications , Humains , Incidence , Infections/complications , Syndrome du côlon irritable/complications , Syndrome du côlon irritable/épidémiologie , Études longitudinales , Mâle , Adulte d'âge moyen , Facteurs de risque , Enquêtes et questionnaires , Jeune adulteRÉSUMÉ
Pulmonary vein deceleration injury is rare and patients can be deceptively stable for a period after injury. Quick diagnosis and transfer to the operating theatre is the only way to treat this potentially lethal injury successfully. Techniques of repair are a useful addition to the cardiovascular surgeon's repertoire.
Sujet(s)
Veines pulmonaires/traumatismes , Blessures du thorax/chirurgie , Thoracotomie/méthodes , Procédures de chirurgie vasculaire/méthodes , Lésions du système vasculaire/chirurgie , Plaies non pénétrantes/chirurgie , Humains , Mâle , Veines pulmonaires/chirurgie , Blessures du thorax/diagnostic , Tomodensitométrie , Indices de gravité des traumatismes , Lésions du système vasculaire/diagnostic , Plaies non pénétrantes/diagnostic , Jeune adulteRÉSUMÉ
Prophylaxis for surgical site infection (SSI) is often at variance with guidelines, despite the prevalence of SSI and its associated cost, morbidity, and mortality. The CareTrack Australia study, undertaken by a number of the authors, demonstrated that appropriate care (in line with evidence- or consensus-based guidelines) was provided at 38% of eligible SSI healthcare encounters. Here, we report the indicator-level CareTrack Australia findings for SSI prophylaxis. Indicators were extracted from Australian and international clinical guidelines and ratified by clinical experts. A sample designed to be representative of the Australian population was recruited (n=1154). Participants' medical records were reviewed and analysed for compliance with the five SSI indicators. The main outcome measure was the percentage of eligible healthcare encounters with documented compliance with indicators for appropriate SSI prophylaxis. Of the 35,145 CareTrack Australia encounters, 702 (2%) were eligible for scoring against the SSI indicators. Where antibiotics were recommended, compliance was 49% for contaminated surgery, 57% for clean-contaminated surgery and 85% for surgery involving a prosthesis: these fell to 8%, 10% and 14%, respectively (an average of 11%), when currently recommended timing of antibiotic administration was included. Where antibiotics were not indicated, 72% of patients still received them. SSI prophylaxis in our sample was poor; over two-thirds of patients were given antibiotics, whether indicated or not, mainly at the wrong time. There is a need for national agreement on clinical standards, indicators and tools to guide, document and monitor SSI prophylaxis, with both local and national measures to increase and monitor their uptake.
Sujet(s)
Antibioprophylaxie/méthodes , Antibioprophylaxie/statistiques et données numériques , Adhésion aux directives/statistiques et données numériques , Enquêtes sur les soins de santé/méthodes , Infection de plaie opératoire/prévention et contrôle , Adolescent , Adulte , Sujet âgé , Australie , Femelle , Enquêtes sur les soins de santé/statistiques et données numériques , Humains , Mâle , Dossiers médicaux , Adulte d'âge moyen , Jeune adulteRÉSUMÉ
High altitude exposure normally leads to a marked natriuresis and diuresis. Acute mountain sickness is often associated with fluid retention, to which an elevated cortisol may contribute. Most investigators report a rise in resting cortisol with ascent, but little data exist regarding the cortisol response to a day trekking. We therefore measured salivary cortisol during ascent to > 5000 m in a cohort of between 42-45 subjects following a 6-h trek (samples taken between 15:30-16:30 h) and between 15-20 subjects at rest (morning samples taken between 08:00-09:00 h). Morning resting cortisol [nmol/l, mean±sd, (range)] was 5.5±2.9 (2.13-13.61) at 1300 m; 4.7±6.8 (1.4-27.02) at 3400 m, and significantly (p=0.002) rose between 4270 m [3.5±2.1 (1.4-8.34)] and 5150 m [14.5±30.3 (1.9-123.1)]. Post-exercise cortisol [nmol/l, mean±sd, (range)] dropped between 3400 m [7±6 (1.5-33.3)] and 4270 m [4.2±4.8 (1.4-29.5)] (p=0.001) followed by a significant rise in post-exercise cortisol between 4270 m [4.2±4.8 (1.4-29.5)] and 5 150 m [9.2±10.2 (1.4-61.3)] (p<0.001). There were no significant associations between severity of acute mountain sickness and cortisol levels. There was a significant though weak correlation between cortisol post-exercise at 5150 m and oxygen saturation at 5150 m (rho= - 0.451, p=0.004). In conclusion, this is the largest cohort to have their resting and post-exercise cortisol levels ascertained at high altitude. We confirm the previous findings of an elevated resting morning cortisol at > 5000 m, but present the novel finding that the cortisol response to a day trekking at HA appears suppressed at 4270 m.
Sujet(s)
Mal de l'altitude/métabolisme , Exercice physique/physiologie , Hydrocortisone/métabolisme , Hypoxie/métabolisme , Repos/physiologie , Adulte , Femelle , Humains , Hydrocortisone/analyse , Mâle , Adulte d'âge moyen , Salive/composition chimique , Salive/métabolisme , Jeune adulteRÉSUMÉ
AIM: To examine the response of brain natriuretic peptide (BNP) and NT-proBNP to high altitude (HA) both at rest and following exercise. METHODS: We measured NT-proBNP and BNP and Lake Louise (LL) acute mountain sickness (AMS) scores in 20 subjects at rest in Kathmandu (Kat; 1300 m), following exercise and at rest at 4270 and 5150 m. RESULTS: BNP and NT-proBNP (pg ml(-1) , mean ± SEM) rose significantly from Kat (9.2 ± 2 and 36.9 ± 6.6, respectively) to arrival at 4270 m after exercise (16.6 ± 4 and 152 ± 56.1, P=0.008 and P<0.001, respectively) and remained elevated the next morning at rest (28.9 ± 9 and 207.4 ± 65.1, P = 0.004 and P<0.001 respectively). At 5150, immediately following ascent/descent to 5643 m, BNP and NT-proBNP were 32.3 ± 8.8 and 301.1 ± 96.3 (P=0.003 and P<0.001 vs. Kat, respectively) and at rest the following morning were 33.3 ± 9.7 and 258.9 ± 89.5 (P=0.008 and P=0.001 vs. Kat respectively). NT-proBNP and BNP correlated strongly at 5150 m (ρ 0.905, P<0.001 and ρ 0.914, P<0.001 for resting and post-exercise samples respectively). At 5150 m, BNP levels were significantly higher among the four subjects with severe (LL score>6) AMS (58.4 ± 18.7) compared with those without (BNP 22.7 ± 8.6, P=0.048). There were significant correlations between change in body water from baseline to 5150 m with both BNP and NT-proBNP (ρ 0.77, P=0.001, ρ 0.745, P=0.002 respectively). CONCLUSION: In conclusion, these data suggest that BNP and NT-proBNP increase with ascent to HA both after exercise and at rest. We also report the novel finding that BNP is significantly greater in those with severe AMS at 5150 m.
Sujet(s)
Mal de l'altitude/sang , Peptide natriurétique cérébral/sang , Fragments peptidiques/sang , Maladie aigüe , Adulte , Altitude , Marqueurs biologiques/sang , Exercice physique/physiologie , Femelle , Humains , Mâle , Adulte d'âge moyen , Népal , Repos/physiologieRÉSUMÉ
The role of the cardiovascular system is to deliver oxygenated blood to the tissues and remove metabolic effluent. It is clear that this complex system will have to adapt to maintain oxygen deliver in the profound hypoxia of high altitude. The literature on the adaptation of both the systemic and pulmonary circulations to high altitude is reviewed.
Sujet(s)
Mal de l'altitude/physiopathologie , Altitude , Phénomènes physiologiques cardiovasculaires , Alpinisme/physiologie , Pression sanguine , Débit cardiaque , Rythme cardiaque , Humains , Hypertension pulmonaire/étiologie , Hypertension pulmonaire/physiopathologie , Effort physique/physiologie , Volume plasmatique , Circulation pulmonaireRÉSUMÉ
INTRODUCTION: The incidence of Acute Mountain Sickness (AMS) is increasing. In a military context our current operational areas include mountainous regions with the implications of AMS including loss of operational tempo and logistical overstretch. Oxygen saturation and heart rate variability have in some studies been predictive of AMS while in others not. No single factor has been demonstrated consistently to be predictive of developing AMS. METHODS: During an expedition to climb Mt Aconcagua (6959m) we explored the relationship between cardiorespiratory variables and AMS. In 11 subjects we measured simple physiological variables and Lake Louise Score both pre and post a standardised exercise challenge at on arrival at different altitudes and after a period of acclimatization. RESULTS: The changes in cardiorespiratory variables we observed with altitude were consistent with previous studies. Heart rate, respiratory rate and blood pressure increased whilst oxygen saturation reduced. Over time at altitude, respiratory rate and heart rate were maintained whilst there was a reduction in blood pressure towards sea level values. Oxygen saturations improved over time at altitude and the change in heart rate on exercise was reduced with acclimatization. In this small pilot study individuals with AMS may have a greater heart rate response to exercise than non-AMS subjects and this may warrant further investigation. CONCLUSIONS: The incidence of AMS in our study was low reflecting a conservative ascent profile. Further larger studies are necessary to fully assess the predictive value of cardiorespiratory variables in AMS.
Sujet(s)
Mal de l'altitude/physiopathologie , Phénomènes physiologiques cardiovasculaires , Personnel militaire , Maladie aigüe , Adulte , Mal de l'altitude/épidémiologie , Argentine , Pression sanguine/physiologie , Femelle , Rythme cardiaque/physiologie , Humains , Incidence , Mâle , Adulte d'âge moyen , Consommation d'oxygène/physiologie , Projets pilotes , Fréquence respiratoire/physiologie , Royaume-UniRÉSUMÉ
Given the heterogeneous nature of multiple sclerosis (MS), we applied DNA microarray technology to determine whether variability is reflected in peripheral blood (PB) cells. In this study, we studied whole-blood gene expression profiles of 29 patients with relapsing-remitting MS (RRMS) and 25 age- and sex-matched healthy controls. We used microarrays with a complexity of 43K cDNAs. The data were analyzed using sophisticated pathway-level analysis in order to provide insight into the deregulated peripheral immune response programs in MS. We found a remarkable elevated expression of a spectrum of genes known to be involved in immune defense in the PB of MS patients compared to healthy individuals. Cluster analysis revealed that the increased expression of these genes was characteristic for approximately half of the patients. In addition, the gene signature in this group of patients was comparable with a virus response program. We conclude that the transcriptional signature of the PB cells reflects the heterogeneity of MS and defines a sub-population of RRMS patients, who exhibit an activated immune defense program that resembles a virus response program, which is supportive for a link between viruses and MS.
Sujet(s)
Sclérose en plaques récurrente-rémittente/génétique , Sclérose en plaques récurrente-rémittente/immunologie , Études cas-témoins , Analyse de regroupements , Régulation de l'expression des gènes , Hétérogénéité génétique , Humains , Interféron de type I/immunologie , Interféron de type I/métabolisme , Sclérose en plaques récurrente-rémittente/sang , Séquençage par oligonucléotides en batterie , Infections à Poxviridae/génétique , Transduction du signal , Régulation positiveRÉSUMÉ
The objective of the present study was to develop a multiplex polymerase chain reaction (PCR) method for differential detection of turkey coronavirus (TCoV), infectious bronchitis coronavirus (IBV), and bovine coronavirus (BCoV). Primers were designed from conserved or variable regions of nucleocapsid (N) or spike (S) protein gene among TCoV, IBV, and BCoV and used in the same PCR reaction. Reverse transcription followed by the PCR reaction was used to amplify a portion of N or S gene of the corresponding coronaviruses. The PCR products were detected on agarose gel stained with ethidium bromide. Two PCR products, a 356-bp band corresponding to N gene and a 727-bp band corresponding to S gene, were obtained for TCoV isolates. In contrast, one PCR product of 356 bp corresponding to a fragment of N gene was obtained for IBV strains and one PCR product of 568 bp corresponding to a fragment of S gene was obtained for BCoV. There were no PCR products with the same primers for Newcastle disease virus, Marek's disease virus, turkey pox virus, pigeon pox virus, fowl pox virus, reovirus, infectious bursal disease virus, enterovirus, astrovirus, Salmonella enterica, Escherichia coli, and Mycoplasma gallisepticum. Performance of the assay with serially diluted RNA demonstrated that the multiplex PCR could detect 4.8x10(-3) microg of TCoV RNA, 4.6x10(-4) microg of IBV RNA, and 8.0x10(-2) microg of BCoV RNA. These results indicated that the multiplex PCR as established in the present study is a rapid, sensitive, and specific method for differential detection of TCoV, IBV, and BCoV in a single PCR reaction.
Sujet(s)
Coronavirus bovin/génétique , Coronavirus de la dinde/génétique , Virus de la bronchite infectieuse/génétique , Réaction de polymérisation en chaîne/méthodes , Animaux , Infections à coronavirus/diagnostic , Protéines de la nucléocapside des coronavirus , Coronavirus bovin/composition chimique , Coronavirus de la dinde/composition chimique , Amorces ADN , Gènes viraux , Virus de la bronchite infectieuse/composition chimique , Glycoprotéines membranaires/génétique , Protéines nucléocapside/génétique , ARN viral/génétique , Sensibilité et spécificité , Spécificité d'espèce , Glycoprotéine de spicule des coronavirus , Dindons , Protéines de l'enveloppe virale/génétiqueRÉSUMÉ
OBJECTIVES: To define best practice standards for mitral valve repair surgery. DESIGN: Development of standards for process and outcome by consensus. SETTING: Multidisciplinary panel of surgeons, anaesthetists, and cardiologists with interests and expertise in caring for patients with severe mitral regurgitation. MAIN OUTCOME MEASURES: Standards for best practice were defined including the full spectrum of multidisciplinary aspects of care. RESULTS: 19 criteria for best practice were defined including recommendations on surgical training, intraoperative transoesophageal echocardiography, surgery for atrial fibrillation, audit, and cardiology and imaging issues. CONCLUSIONS: Standards for best practice in mitral valve repair were defined by multidisciplinary consensus. This study gives centres undertaking mitral valve repair an opportunity to benchmark their care against agreed standards that are challenging but achievable. Working towards these standards should act as a stimulus towards improvements in care.
Sujet(s)
Insuffisance mitrale/chirurgie , Pratique professionnelle/normes , Fibrillation auriculaire/chirurgie , Cardiologie/enseignement et éducation , Cardiologie/normes , Consultants , Échocardiographie transoesophagienne , Formation médicale continue comme sujet , Chirurgie générale/enseignement et éducation , Chirurgie générale/normes , Hôpitaux/normes , Hôpitaux/statistiques et données numériques , Humains , Soins peropératoires , Audit médical , Personnel médical hospitalier/normes , Personnel médical hospitalier/statistiques et données numériques , Équipe soignante , Normes de référence , Procédures de chirurgie thoracique/normes , Procédures de chirurgie thoracique/statistiques et données numériques , Royaume-UniRÉSUMÉ
An experimental system of Mycobacterium tuberculosis growth in a carbon-limited chemostat has been established by the use of Mycobacterium bovis BCG as a model organism. For this model, carbon-limited chemostats with low concentrations of glycerol were used to simulate possible growth rates during different stages of tuberculosis. A doubling time of 23 h (D = 0.03 h(-1)) was adopted to represent cells during the acute phase of infection, whereas a lower dilution rate equivalent to a doubling time of 69 h (D = 0.01 h(-1)) was used to model mycobacterial persistence. This chemostat model allowed the specific response of the mycobacterial cell to carbon limitation at different growth rates to be elucidated. The macromolecular (RNA, DNA, carbohydrate, and lipid) and elemental (C, H, and N) compositions of the biomass were determined for steady-state cultures, revealing that carbohydrates and lipids comprised more than half of the dry mass of the BCG cell, with only a quarter of the dry weight consisting of protein and RNA. Consistent with studies of other bacteria, the specific growth rate impacts on the macromolecular content of BCG and the proportions of lipid, RNA, and protein increased significantly with the growth rate. The correlation of RNA content with the growth rate indicates that ribosome production in carbon-limited M. bovis BCG cells is subject to growth rate-dependent control. The results also clearly show that the proportion of lipids in the mycobacterial cell is very sensitive to changes in the growth rate, probably reflecting changes in the amounts of storage lipids. Finally, this study demonstrates the utility of the chemostat model of mycobacterial growth for functional genomic, physiology, and systems biology studies.
Sujet(s)
Métabolisme lipidique , Mycobacterium bovis/croissance et développement , Mycobacterium bovis/métabolisme , Ribosomes/métabolisme , Milieux de culture , Structures macromoléculaires/métabolismeRÉSUMÉ
BACKGROUND: Atrial en-face reconstructions are commonly used to assess mitral valve morphology in three dimensional (3D) echocardiography but may miss important abnormalities. OBJECTIVE: To present a systematic method for the analysis of the regurgitant mitral valve using a combination of en-face and longitudinal views for better anatomical evaluation. METHODS: Detailed 3D assessment was done on 58 patients undergoing mitral valve repair. En-face and longitudinal views were compared for detection and location of primary pathology. The quality of acquisitions under general anaesthesia and sedation was also compared. RESULTS: Recognition of valve structure was significantly better with longitudinal reconstruction for both mitral leaflets but not for the commissures. Accurate identification of pathology was possible in 95% cases, compared with 50% for en-face reconstruction (p < 0.001). There was no significant difference between imaging under sedation and anaesthesia. CONCLUSION: En-face reconstructions alone are inadequate. Additional longitudinal reconstructions are necessary to ensure full inspection of valve morphology.
Sujet(s)
Échocardiographie tridimensionnelle/méthodes , Insuffisance mitrale/imagerie diagnostique , Adulte , Sujet âgé , Anesthésie générale , Sédation consciente , Femelle , Humains , Mâle , Adulte d'âge moyen , Insuffisance mitrale/étiologie , Biais de l'observateur , Facteurs tempsRÉSUMÉ
Purification of turkey coronavirus (TCoV) nucleocapsid (N) protein, expressed in a prokaryotic expression system as histidine-tagged fusion protein is demonstrated in the present study. Turkey coronavirus was partially purified from infected intestine of turkey embryo by sucrose gradient ultracentrifugation and RNA was extracted. The N protein gene was amplified from the extracted RNA by reverse transcription-polymerase chain reaction and cloned. The recombinant expression construct (pTri-N) was identified by polymerase chain reaction and sequencing analysis. Expression of histidine-tagged fusion N protein with a molecular mass of 57 kd was determined by Western blotting analysis. By chromatography on nickel-agarose column, the expressed N protein was purified to near homogeneity as judged by sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis. The protein recovery could be 2.5 mg from 100 ml of bacterial culture. The purified N protein was recognized by antibody to TCoV in Western blotting assay. The capability of the recombinant N protein to differentiate positive serum of turkey infected with TCoV from normal turkey serum was evident in enzyme-linked immunosorbent assays (ELISA). These results indicated that the expressed N protein is a superior source of TCoV antigen for development of antibody-capture ELISA for detection of antibodies to TCoV.
Sujet(s)
Coronavirus de la dinde/croissance et développement , Coronavirus de la dinde/isolement et purification , Escherichia coli/virologie , Protéines nucléocapside/isolement et purification , Animaux , Séquence nucléotidique , Amorces ADN , Embryon non mammalien/virologie , Entérite transmissible du dindon/embryologie , Entérite transmissible du dindon/virologie , Test ELISA/méthodes , Protéines nucléocapside/génétique , ARN viral/isolement et purification , Dindons/embryologieRÉSUMÉ
The objective of this study was to elucidate the kinetics and magnitudes of specific IgA antibody responses in intestines of turkey poults infected with turkey coronavirus (TCV). Turkey poults were orally inoculated with TCV at 10 days of age. Intestinal segment cultures were administered for duodenum, jejunum, and ileum and the IgA antibody responses were analyzed at 1, 2, 3, 4, 6, or 9 weeks post-infection (PI) in two different experiments. The kinetics of virus-specific IgA antibody responses in duodenum, jejunum, and ileum were similar: gradually increased from 1 week PI, reached the peak at 3 or 4 weeks PI, and declined afterward. The virus-specific IgA antibody responses in duodenum, jejunum, and ileum showed negative correlation with duration of TCV antigen in the corresponding locations of intestine with Spearman's correlation coefficient of -0.85 (p=0.034), -0.74 (p=0.096), and -0.75 (p=0.084), respectively. Moreover, the virus-specific IgA antibody responses in serum were positively correlated with that of duodenum (coefficient=0.829, p=0.042), jejunum (coefficient=0.829, p=0.042), and ileum (coefficient=0.771, p=0.072) segment cultures, suggesting that the induction of specific IgA response in serum was predictive of an IgA response in intestine. The results indicate that intestinal mucosal IgA antibodies to TCV are elicited in turkeys following infection with TCV. The local mucosal antibodies may provide protective immunity for infected turkeys to recover from TCV infection.
Sujet(s)
Coronavirus/immunologie , Entérite transmissible du dindon/immunologie , Immunoglobuline A/immunologie , Intestin grêle/immunologie , Maladies de la volaille/immunologie , Dindons , Animaux , Spécificité des anticorps , Antigènes viraux/immunologie , Entérite transmissible du dindon/virologie , Test ELISA/médecine vétérinaire , Technique d'immunofluorescence directe/médecine vétérinaire , Immunité muqueuse/immunologie , Immunoglobuline A/biosynthèse , Muqueuse intestinale/immunologie , Muqueuse intestinale/ultrastructure , Muqueuse intestinale/virologie , Intestin grêle/ultrastructure , Intestin grêle/virologie , Cinétique , Microscopie électronique , Maladies de la volaille/virologie , Organismes exempts d'organismes pathogènes spécifiquesRÉSUMÉ
Sephacryl S-1000 size-exclusion chromatography was used to purify turkey coronavirus (TCoV) from infected turkey embryo. TCoV was propagated in the 22-day-old turkey embryos. Intestines and intestinal contents of infected embryos were harvested and homogenized. After low speed centrifugation, the supernatant was concentrated by ultracentrifugation through a cushion of 30 or 60% sucrose solution, or by ammonium sulfate precipitation. The purification methods included sucrose gradient and Sephacryl S-1000 size-exclusion chromatography. Ultracentrifugation through a cushion of 60% sucrose solution was better than the other two methods for concentration of TCoV from intestinal homogenate. The most effective method for purifying TCoV and removing extraneous materials was size-exclusion chromatography as analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. More spike-rich particles were observed in the sample purified by chromatography than those purified by sucrose gradient as examined by electron microscopy. Differentiation of turkey anti-TCoV antiserum from normal turkey serum was better achieved by ELISA plates coated with TCoV preparation purified by size-exclusion chromatography than that purified by sucrose density gradient. The results indicated that Sephacryl S-1000 chromatography was useful for purification of TCoV.