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AIMS: To evaluate the organ at risk (OAR) dose and its characteristic variation with different clinically usable prescription doses (RxD) for breast and chest wall radiotherapy in patients with early-stage left-sided breast cancer. MATERIALS AND METHODS: In total, 145 patients with early-stage breast cancers (T1N0M0-T2N0M0) on the left side were treated with radiotherapy after a modified radical mastectomy or breast conservation surgery, with a mean age of 45.1 ± 21.6 years. The patient received 4050 cGy of field-in-field (three-dimensional conformal radiotherapy) treatment limited to the breast or chest wall, excluding the supraclavicular node, axillary node and internal mammary chain, over 15 fractions. Additional plans of 5000 cGy/25 fractions, 4500 cGy/20 fractions and 2600 cGy/5 fractions were created with no or minor changes to the original plan. Mathematical modelling was used to study the distinctive change in the dose-volume characteristics for various OARs as a function of the RxD. OAR dosages, both absolute and normalised, were expressed in terms of the RxD. The mathematical (functional) relationship between OAR doses and different prescription levels was deduced by the least squares fit method. RESULT: The left lung mean dose, V5Gy (%), V10Gy (%) and V20Gy (%) and the heart mean dose, V10Gy (%) and V20Gy (%) were evaluated. The dose-volume parameters showed a parabolic variation (x2) with the RxD. Prescription normalised OAR doses showed a linear relationship with the RxD; relative dose increased with diminishing RxD. Normalised lung and heart mean doses exhibited saturation (linear relationship) with RxD variation. Paired sample t-test results between RxD versus all evaluated parameters were found to be statistically significant (P = 0.004). The Pearson correlation coefficient between different prescription levels for left lung mean dose (range 0.942-1.0), heart mean dose (range 1.0-1.0), left lung V5Gy (%) (range 0.987-1.0), left lung V10Gy (%) (range 0.991-0.999), heart V10Gy (%) (range 0.998-1.0). CONCLUSION: The functional form of absolute OAR dose-volume parameters versus RxD is parabolic and the RxD normalised OAR dose-volume parameter versus RxD is a straight line with a negative slope as RxD increases. This indicates an increase in the relative OAR dose-volume parameters if the RxD is reduced. This study is the first of its kind to compare the OAR doses as a function of clinically used degenerate prescription levels. These data will help to comprehend the OAR doses while adopting a new dose fractionation regimen and reviewing the radiotherapy treatment plans.
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Neoplasias da Mama , Radioterapia de Intensidade Modulada , Neoplasias Unilaterais da Mama , Humanos , Adulto Jovem , Adulto , Pessoa de Meia-Idade , Idoso , Feminino , Neoplasias da Mama/radioterapia , Neoplasias da Mama/cirurgia , Mastectomia , Dosagem Radioterapêutica , Planejamento da Radioterapia Assistida por Computador/métodos , Órgãos em Risco , Radioterapia de Intensidade Modulada/métodosRESUMO
To assess BAT activity in humans at a population level, infrared thermography (IRT) represents a safe, readily repeatable and affordable alternative to 18F-FDG-PET. Building upon a previously proposed method by our laboratory, we further refined the image computational algorithm to quantify BAT activation in the cervical-supraclavicular (C-SCV) region of healthy young men under thermo-neutral and cold exposure conditions. Additionally, we validated the whole-body calorimeter (WBC) in reliably measuring cold-induced thermogenesis. The temperature gradient between C-SCV-deltoid regions, and the corresponding difference in heat power output, increased upon cold air exposure relative to thermo-neutral conditions (by 74.88 %, p<0.0001; and by 71.34 %, p<0.0001 respectively). Resting and cumulative energy expenditure (EE) rose significantly (by 13.14 % and 9.12 % respectively, p=0.0001) while positive correlations between IRT measures and EE were found with cold air exposure (percentage change in heat power gradient between ROI and deltoid, cold air: r(2)=0.29, p=0.026, Pearson's correlation). IRT and WBC can be used to study BAT activation. The refined algorithm allows for more automation and objectivity in IRT data analysis, especially under cold air exposures.
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Tecido Adiposo Marrom/metabolismo , Termogênese , Adulto , Calorimetria Indireta , Humanos , Masculino , Termografia , Adulto JovemRESUMO
Effect of charge carrier confinement and ultra-low disorder acquainted in AlGaAs/GaAs multi-quantum well system is investigated via Magneto-photoluminescence spectroscopy. Significant increase of effective mass is observed for the confined exciton in narrow QWs. The foremost reason behind such an observation is due to the induced non-parabolicity in bands. Moreover, as the thickness of the QW are reduced, confined excitons in QW experience atomic irregularities at the hetero-junctions and their effects are prominent in the photoluminescence linewidth. Amount of photoluminescence line-broadening caused by the atomic irregularities at the hetero-junctions is correlated with average fluctuation (δ 1) in QW thickness. The estimated δ 1 for Al0.3Ga0.7As/GaAs QWs are found to be ±(0.14 - 1.6)× 'one monolayer thickness of GaAs layer'. Further, the strong perturbations due to magnetic field in a system helps in realizing optical properties of exciton in QWs, where magnetic field is used as a probe to detect ultralow defects in the QW. Additionally, the influence of magnetic field on the free and bound exciton luminescence is explained by a simple model. The proposed approach for measuring the interface and volume defects in an ultra-low disordered system by Magneto-PL spectroscopy technique will be highly beneficial in high mobility devices for advanced applications.
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The novel opto-chemical doping effect in Ag nanoparticle-decorated monolayer graphene grown by chemical vapor deposition has been investigated using Raman spectroscopy for the first time. We used both noble metal nanoparticles and optical excitation, in a hybrid opto-chemical route, to tune the doping level in graphene. Metal nanoparticle-induced chemical effects and laser power-induced substrate effects alter the doping nature of graphene from p- to n-type. Compared with earlier studies, the proposed method significantly lowers the laser intensity required for optical power-dependent doping, resulting in prevention of damage to the sample due to local heating. Some other interesting observations are the enhanced peak intensity in the Raman spectrum of graphene, enhancement of the D-band intensity and the introduction of G-band splitting. This novel, cheap and easily implemented hybrid optical-chemical doping strategy could be very useful for tuning graphene plasmons on the widely used Si/SiO2 substrates for various photonic device applications.
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INTRODUCTION: Sputum culture conversion in pulmonary multidrug-resistant tuberculosis (MDR-TB) is important to make treatment-related decisions and prevent transmission of disease. OBJECTIVE: To identify factors associated with sputum culture conversion, and to determine time to culture conversion and the impact of culture conversion on successful treatment outcomes in MDR-/rifampicin (RMP) resistant TB. METHOD: Retrospective analysis of data from treatment cards and registers of MDR-/RMP-resistant patients initiated on treatment under India's Revised National TB Control Programme in Delhi, West Bengal and Kerala from January 2009 to December 2011. Proportions were calculated and logistic regression analysis was performed. RESULTS: Of 836 patients, 787 were analysed, 651 (83%) of whom experienced culture conversion: respectively 57%, 73% and 79% culture converted by month 3, 4 and 6 of treatment. The median time to culture conversion was 91.3 days. Patients with body mass index (BMI) 16 kg/m2 (OR 0.403, P = 0.001) and 1618 kg/m2 (OR 0.519, P = 0.039) were less likely to have culture conversion. High rates of culture conversion were observed in patients with successful treatment outcomes compared to those without treatment success (462/469, 99% vs. 183/311, 59%; P 0.0001). CONCLUSION: Low BMI is associated with poor sputum culture conversion in MDR-/RMP-resistant TB patients. Lack of culture conversion can impact successful treatment outcomes.
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Escarro/microbiologia , Tuberculose Resistente a Múltiplos Medicamentos/diagnóstico , Tuberculose Resistente a Múltiplos Medicamentos/tratamento farmacológico , Adolescente , Adulto , Antituberculosos/uso terapêutico , Índice de Massa Corporal , Testes Diagnósticos de Rotina , Gerenciamento Clínico , Farmacorresistência Bacteriana Múltipla , Feminino , Humanos , Índia , Masculino , Pessoa de Meia-Idade , Estudos Retrospectivos , Rifampina/uso terapêutico , Resultado do Tratamento , Adulto JovemRESUMO
This study aimed to examine the diagnostic yield of fine needle aspiration cytology (FNAC) and ultrasound-guided core needle biopsy (USCB) in the diagnosis of parotid neoplasia. A 16-year retrospective analysis was performed of patients entered into our pathology database with a final diagnosis of parotid neoplasia. FNAC and USCB data were compared to surgical excision where available. One hundred and twenty FNAC, 313 USCB, and 259 surgical specimens were analyzed from 397 patients. Fifty-six percent of FNAC and 4% of USCB were non-diagnostic. One hundred and thirty-two (33%) patients had a final diagnosis made by USCB and did not undergo surgery. Surgery was performed in 257 (65%) patients, 226 (88%) of whom had a preoperative biopsy. Most lesions were benign, but there were 62 parotid and 13 haematological malignancies diagnosed; false-negative results were obtained in three FNAC and two USCB samples. The sensitivity and specificity of FNAC were 70% and 89%, respectively, and for USCB were 93% and 100%, respectively. This study represents the largest series of patients with a parotid neoplasm undergoing USCB for diagnosis. USCB is highly accurate with a low non-diagnostic rate and should be considered an integral part of parotid assessment.
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Biópsia por Agulha Fina , Biópsia com Agulha de Grande Calibre , Biópsia Guiada por Imagem , Neoplasias Parotídeas/diagnóstico , Ultrassonografia de Intervenção , Adulto , Idoso , Idoso de 80 Anos ou mais , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Neoplasias Parotídeas/patologia , Neoplasias Parotídeas/cirurgia , Valor Preditivo dos Testes , Estudos Retrospectivos , Sensibilidade e EspecificidadeRESUMO
The intra-atomic magnetic dipole moment - frequently called ⟨Tz⟩ term - plays an important role in the determination of spin magnetic moments by x-ray absorption spectroscopy for systems with nonspherical spin density distributions. In this work, we present the dipole moment as a sensitive monitor to changes in the electronic structure in the vicinity of a phase transiton. In particular, we studied the dipole moment at the Fe(2+) and Fe(3+) sites of magnetite as an indicator for the Verwey transition by a combination of x-ray magnetic circular dichroism and density functional theory. Our experimental results prove that there exists a local change in the electronic structure at temperatures above the Verwey transition correlated to the known spin reorientation. Furthermore, it is shown that measurement of the dipole moment is a powerful tool to observe this transition in small magnetite nanoparticles for which it is usually screened by blocking effects in classical magnetometry.
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Serine protease autotransporters of Enterobacteriaceae (SPATEs) are secreted proteins demonstrating diverse virulence functions. The distribution of SPATEs is studied among diarrheagenic and extraintestinal pathogenic Escherichia coli. However, the contribution of SPATEs to the virulence of neonatal septicemic Escherichia coli (NSEC) has not yet been elucidated. This study was undertaken to evaluate the prevalence and phylogenetic distribution of different subtypes of SPATEs among NSEC. The presence of virulence factors and subtypes of SPATEs among different E. coli isolates was determined by polymerase chain reaction (PCR). E. coli phylogrouping was done by triplex PCR. Clonality of the isolates was assessed by pulsed-field gel electrophoresis (PFGE). The presence of SPATEs was significantly higher among the septicemic isolates (89 %) than the fecal (7.5 %) and environmental isolates (2.5 %). Vat (vacuolating autotransporter toxin) and Sat (secreted autotransporter toxin) were found to be the two most predominant SPATEs. The incidence of SPATEs was high in septicemic isolates of phylogroups A and B1 (87 %), lacking other virulence factors. The high prevalence of SPATEs in the non-B2 phylogroups of septicemic isolates in comparison with fecal and environmental isolates indicates an association of SPATEs with NSEC. The NSEC isolates were found to be clonally distinct, suggesting that the high prevalence of SPATEs was not due to clonal relatedness of the isolates. This study is the first to show the association of SPATEs with NSEC. The presence of SPATEs in the septicemic/NSEC isolates may be considered as the most discriminatory trait studied here.
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Infecções por Escherichia coli/microbiologia , Escherichia coli/enzimologia , Transporte Proteico , Sepse/microbiologia , Serina Proteases/metabolismo , Análise por Conglomerados , Eletroforese em Gel de Campo Pulsado , Escherichia coli/classificação , Escherichia coli/genética , Escherichia coli/isolamento & purificação , Infecções por Escherichia coli/epidemiologia , Feminino , Genótipo , Humanos , Lactente , Recém-Nascido , Masculino , Tipagem Molecular , Filogenia , Reação em Cadeia da Polimerase , Prevalência , Sepse/epidemiologia , Serina Proteases/classificação , Serina Proteases/genética , Fatores de Virulência/genéticaRESUMO
The aim of this study was to evaluate the performance of fine needle aspiration cytology (FNAC), ultrasound-guided core needle biopsy (USCNB), punch biopsy, and surgical excision biopsy in neoplasms presenting within the submandibular space. A retrospective analysis of all patients with a pathological diagnosis of a submandibular space neoplasm within a 12-year period (February 1999 to June 2011) was performed. Biopsy results were compared to histopathological diagnosis obtained from surgical excision biopsy. Eighty-one specimens from 44 patients met the search criteria (15 FNAC, 24 USCNB, 7 punch biopsy, and 35 surgical excision biopsy). The final diagnosis was established by USCNB, punch biopsy, or surgical excision biopsy and not by FNAC alone. Surgical excision biopsy was performed as a primary diagnostic (n = 8), secondary diagnostic (n = 15), or as a post-diagnostic therapeutic procedure (n = 12). Non-diagnostic results were: FNAC 11/15, USCNB 2/24, and punch biopsy 1/7. Diagnostic results were: FNAC 2/15, USCNB 20/24, and punch biopsy 5/7. No complications were reported. Although punch biopsy demonstrated good yield and accuracy, its use is restricted to a small cohort of patients. USCNB is a safe and accurate technique in the submandibular space, with a low non-diagnostic rate.
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Biópsia/métodos , Neoplasias da Glândula Submandibular/patologia , Adulto , Idoso , Idoso de 80 Anos ou mais , Diagnóstico Diferencial , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Ultrassonografia de IntervençãoAssuntos
Imunossupressores/administração & dosagem , Tacrolimo/administração & dosagem , Terapia Ultravioleta/métodos , Vitiligo/tratamento farmacológico , Vitiligo/radioterapia , Administração Tópica , Adolescente , Adulto , Terapia Combinada/métodos , Feminino , Humanos , Masculino , Resultado do Tratamento , Vitiligo/diagnóstico , Adulto JovemRESUMO
1. In a 42-d feeding trial, 264 one-d-old, as hatched, Cobb 400 broiler chickens (6 pens per group, n = 11 per pen in a 2 × 2 factorial arrangement) were fed on two concentrations of dietary calcium (Ca) (9.0 and 7.5 g/kg in starter, 7.5 and 6 g/kg in grower phases) and supplemental phytase (0 and 500 U/kg diet). 2. During d 0-21, the high Ca + phytase diet improved body weight. During d 0-42, feed intake was increased by the low Ca diet and decreased by phytase supplementation. Feed conversion ratio during d 0-21 was improved by the high Ca + phytase diet. 3. At d 42, Ca in duodenal digesta was reduced by low dietary Ca and supplemental phytase. High dietary Ca reduced P in duodenal and jejunal digesta. Phytase reduced digesta P and increased serum P concentration. 4. Relative tibia length decreased with low dietary Ca and increased with phytase. The robusticity index of tibia was improved by the low Ca diet and phytase supplementation. Phytase supplementation increased tibia ash and concentrations of Ca, magnesium (Mg), manganese (Mn), copper (Cu), zinc (Zn) and iron (Fe) in tibia. The low Ca diet increased Mg, Mn and Fe and reduced Cu and Zn in tibia. 5. It was concluded that 7.5 g Ca/kg during weeks 0-3 and 6 g Ca/kg during weeks 3-6 sustained broiler performance and bone ash, while phytase supplementation facilitated tibia mineralisation, particularly during the grower phase.
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6-Fitase/administração & dosagem , Calcificação Fisiológica , Cálcio da Dieta/metabolismo , Galinhas/fisiologia , Tíbia/fisiologia , Ração Animal/análise , Criação de Animais Domésticos , Fenômenos Fisiológicos da Nutrição Animal , Animais , Cálcio da Dieta/sangue , Galinhas/crescimento & desenvolvimento , Dieta/veterinária , Suplementos Nutricionais/análise , Relação Dose-Resposta a Droga , Feminino , Conteúdo Gastrointestinal/química , Intestino Delgado/química , Masculino , Minerais/metabolismo , Distribuição Aleatória , Espectrofotometria Atômica/veterinária , Tíbia/anatomia & histologia , Aumento de Peso/efeitos dos fármacosRESUMO
Graphene is a two-dimensional material with a capability of gas sensing, which is here shown to be drastically improved by inducing gentle disorder in the lattice. We report that by using a focused ion beam technique, controlled disorder can be introduced into the graphene structure through Ga(+) ion irradiation. This disorder leads to an increase in the electrical response of graphene to NO(2) gas molecules by a factor of three in an ambient environment (air). Ab initio density functional calculations indicate that NO(2) molecules bind strongly to Stone-Wales defects, where they modify electronic states close to the Fermi level, which in turn influence the transport properties. The demonstrated gas sensor, utilizing structurally defected graphene, shows faster response, higher conductivity changes and thus higher sensitivity to NO(2) as compared to pristine graphene.
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The study compared the effects of an antibiotic growth promoter (AGP), yeast (Saccharomyces cerevisiae) and yeast cell wall (YCW) on performance, microbiology and histo-morphology of the small intestine and humoral immune responses in Ross 308 broilers. The treatments (eight replicates/treatment, n = 12/replicate) were negative control (NC, without AGP), positive control (PC, supplemented with bacitracin methylene disalicylate, 400 mg/kg), Y and YCW (supplemented with yeast and YCW, respectively, 1000 mg/kg). Live weight at 42 days improved (p = 0.086) in the PC, Y and YCW groups. Feed conversion ratio was better (p = 0.039) in the YCW group compared with the other groups. Antibiotic growth promoter in the PC group shortened the villi in duodenum (p = 0.044). Mucosal Escherichia coli number was higher in the PC group (p < 0.001), whereas in the digesta E. coli number was lower (p = 0.001) in the PC, Y and YCW groups in relation to the NC. Mucosal Salmonella populations increased (p = 0.0001) in the PC group, whereas in the digesta, all treatments reduced the Salmonella (p = 0.0001). Following oral challenge with Salmonella pullorum, YCW increased E. coli numbers on the mucosa (p < 0.001) whereas in the digesta the Y group had lower (p < 0.0001) number of E. coli. In the digesta, Salmonella count was lower in the YCW group compared with the other treatments (p < 0.01). Yeast cell wall -treated birds exhibited better (p < 0.05) humoral immune response against Newcastle disease which was far more persistent over time than in any other treatments. It was concluded that the yeast and the yeast cell wall may have effects identical to BMD on performance of broilers and thus may constitute an effective replacement strategy in the dietary regimens for broiler chickens.
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Antibacterianos/farmacologia , Parede Celular/química , Galinhas/crescimento & desenvolvimento , Dieta/veterinária , Intestinos/efeitos dos fármacos , Leveduras , Ração Animal/análise , Fenômenos Fisiológicos da Nutrição Animal , Animais , Anticorpos Antivirais/sangue , Galinhas/sangue , Galinhas/imunologia , Suplementos Nutricionais , Imunidade Humoral , Intestinos/anatomia & histologia , Doença de Newcastle/imunologia , Doença de Newcastle/prevenção & controle , Vacinas Virais/imunologiaRESUMO
1. The effects of enzymatically hydrolysed whole Saccharomyces cerevisiae yeast (HY) and the pellets of yeast cell wall (YCW) on production traits, the microbiology and histo-morphology of the small intestine, and humoral immune responses against Newcastle disease virus (NDV), of Ross 308 broilers were investigated. 2. The control group received a maize-soyabean meal based basal diet for 42 days. In the treated groups the basal diet was supplemented with 1 g/kg of HY and YCW. There were 8 replicate pens per group (n = 12 birds/pen). 3. HY and YCW supplementation improved live weight (P = 0·006) and FCR (P = 0·003) at 42-d as compared with the control group. 4. In the small intestine, Salmonella spp and Escherichia coli numbers were higher (P = 0·01) in the mucosa and lower (P = 0·01) in the digesta of the HY and the YCW fed groups at 25 d of age. Lactobacillus in the duodenal and jejunal digesta was higher (P < 0·05) in the HY and the YCW fed groups as compared with the control. 5. Following oral challenge with Salmonella pullorum, Escherichia coli and Lactobacillus increased (P < 0·05) in the mucosa and decreased in the digesta (P < 0·05) of the HY and YCW supplemented groups, relative to the control. 6. Supplementation of HY and YCW increased villus height in the jejunum (P = 0·02), width of villus in the ileum (P = 0·034) and number of goblet cells in villi of the jejunum (P = 0·006) and ileum (P = 0·01). 7. YCW increased antibody level against NDV at 21 and 42 d of age compared with the control and the HY supplemented diets (P < 0·05). 8. It was concluded that HY and YCW improved growth and feed efficiency in broilers, and considering the improvements in production traits and humoral immune responses, yeast cell wall may be a better dietary tool than the hydrolysed whole yeast cell as a performance enhancer for broilers.
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Parede Celular/imunologia , Galinhas/imunologia , Galinhas/microbiologia , Intestino Delgado/imunologia , Intestino Delgado/microbiologia , Saccharomyces cerevisiae/imunologia , Animais , Anticorpos Antivirais/sangue , Peso Corporal/imunologia , Contagem de Colônia Microbiana/veterinária , Ingestão de Alimentos/imunologia , Histocitoquímica/veterinária , Imunidade Humoral/imunologia , Mucosa Intestinal/imunologia , Mucosa Intestinal/microbiologia , Intestino Delgado/citologia , Intestino Delgado/ultraestruturaRESUMO
AIMS: To develop an effective multiplex PCR for simultaneous and rapid detection of Vibrio cholerae, Vibrio vulnificus and Vibrio parahaemolyticus, the three most important Vibrio species that can cause devastating health hazards among human. METHODS AND RESULTS: Species-specific PCR primers were designed based on toxR gene for V. cholerae and V. parahaemolyticus, and vvhA gene for V. vulnificus. The multiplex PCR was validated with 488 Vibrio strains including 322 V. cholerae, 12 V. vulnificus, and 82 V. parahaemolyticus, 20 other Vibrio species and 17 other bacterial species associated with human diseases. It could detect the three target bacteria without any ambiguity even among closely related species. It showed good efficiency in detection of co-existing target species in the same sample. The detection limit of all the target species was ten cells per PCR tube. CONCLUSIONS: Specificity and sensitivity of the multiplex PCR is 100% each and sufficient for simultaneous detection of these potentially pathogenic Vibrio species in clinical and environmental samples. SIGNIFICANCE AND IMPACT OF THE STUDY: This simple, rapid and cost-effective method can be applicable in a prediction system to prevent disease outbreak by these Vibrio species and can be considered as an effective tool for both epidemiologist and ecologist.
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Técnicas Bacteriológicas/métodos , Reação em Cadeia da Polimerase/métodos , Vibrioses/diagnóstico , Vibrio cholerae/isolamento & purificação , Vibrio parahaemolyticus/isolamento & purificação , Vibrio vulnificus/isolamento & purificação , Proteínas de Bactérias/genética , Proteínas de Ligação a DNA/genética , Humanos , Sensibilidade e Especificidade , Fatores de Transcrição/genética , Vibrio cholerae/genética , Vibrio parahaemolyticus/genética , Vibrio vulnificus/genéticaRESUMO
A bacterial disease was reported from gilthead sea bream (Sparus aurata) within a hatchery environment in Malta. Symptoms included complete erosion of tail, infection in the eye, mucous secretion and frequent mortality. A total of 540 strains were initially isolated in marine agar from different infected body parts and culture water sources. Subsequently 100 isolates were randomly selected, identified biochemically and all were found to be Vibrio harveyi-related organisms; finally from 100 isolates a total of 13 numbers were randomly selected and accurately identified as V. harveyi by 16S rRNA gene sequencing and species-specific PCR. Ribotyping of these strains with HindIII revealed total of six clusters. In vivo challenge study with representative isolates from each cluster proved two clusters each were highly pathogenic, moderately pathogenic and non-pathogenic. All 13 isolates were positive for hemolysin gene, a potential virulence factor. Further analysis revealed probably a single copy of this gene was encoded in all isolates, although not in the same locus in the genome. Although V. harveyi was reported to be an important pathogen for many aquatic organisms, to our knowledge this might be the first report of disease caused by V. harveyi and their systematic study in the sea bream hatchery from Malta.
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Doenças dos Peixes/microbiologia , Dourada/microbiologia , Vibrioses/veterinária , Vibrio/isolamento & purificação , Animais , Aquicultura , Proteínas de Bactérias/genética , Técnicas de Tipagem Bacteriana , Análise por Conglomerados , DNA Bacteriano/química , DNA Bacteriano/genética , DNA Ribossômico/química , DNA Ribossômico/genética , Exsudatos e Transudatos , Olho/patologia , Doenças dos Peixes/mortalidade , Doenças dos Peixes/patologia , Proteínas Hemolisinas/genética , Malta , Dados de Sequência Molecular , Filogenia , RNA Ribossômico 16S/genética , Análise de Sequência de DNA , Cauda/patologia , Vibrio/classificação , Vibrio/genética , Vibrioses/microbiologia , Vibrioses/mortalidade , Vibrioses/patologia , Fatores de Virulência/genéticaRESUMO
AIMS: To develop simple and rapid PCR-fingerprinting methods for Vibrio cholerae O1 (El Tor and classical biotypes) and O139 serogroup strains which cause major cholera epidemics, on the basis of the diversity of superintegron (SI) carried by these strains. METHODS AND RESULTS: PCR-restriction fragment length polymorphism (PCR-RFLP) assay was developed targeting region between integrase gene in the SI and its nearby ORF, followed by BglI digestion. Besides, a V. cholerae repeat-amplified fragment length polymorphism (VCR-AFLP) assay was also developed. In the PCR-RFLP, 94 El Tor, 29 classical and 54 O139 strains produced nine, three and six different DNA fingerprints, respectively. On the other hand, VCR-AFLP distinguished these El Tor, classical and O139 strains into five, nine and two DNA fingerprints, respectively. Combining both assays the El Tor, classical and O139 strains could be differentiated into 11, 10 and seven different types, respectively. In a comparative study, pulsed-field gel electrophoresis (PFGE) showed similar differentiation for El Tor (11 types), but lower discrimination for O139 (two types) and classical strains (five types). CONCLUSIONS: The PCR assays based on SI diversity can be used as a useful typing tool for epidemiological studies of V. cholerae. SIGNIFICANCE AND IMPACT OF STUDY: This newly developed method is more discriminatory, simple, rapid and cost-effective in comparison with PFGE, and thus can be widely applicable.
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Técnicas de Tipagem Bacteriana/métodos , Variação Genética , Integrons , Reação em Cadeia da Polimerase/métodos , Vibrio cholerae/classificação , Análise do Polimorfismo de Comprimento de Fragmentos Amplificados , Impressões Digitais de DNA/métodos , DNA Bacteriano/análise , Eletroforese em Gel de Campo Pulsado , Polimorfismo de Fragmento de Restrição , Análise de Sequência de DNA , Vibrio cholerae/genéticaRESUMO
AIM: To develop a haemolysin (hly) gene-based species-specific multiplex PCR for simple and rapid detection of Vibrio campbellii, V. harveyi and V. parahaemolyticus. METHODS AND RESULTS: The complete hly genes of three V. campbellii strains isolated from diseased shrimps were sequenced and species-specific PCR primers were designed based on these sequences and the registered hly gene sequences of Vibrio harveyi and Vibrio parahaemolyticus. Specificity and sensitivity of the multiplex PCR was validated with 27 V. campbellii, 16 V. harveyi, and 69 V. parahaemolyticus, 18 other Vibrio species, one Photobacterium damselae and nine other bacterial species. The detection limits of all the three target species were in between 10 and 100 cells per PCR tube. CONCLUSIONS: Specificity and sensitivity of the multiplex PCR is 100% each and sufficient to be considered as an effective tool in a prediction system to prevent potential disease outbreak by these Vibrio species. SIGNIFICANCE AND IMPACT OF THE STUDY: Because there is lack of simple, rapid and cost-effective method to differentiate these closely related V. campbellii, V. harveyi and V. parahaemolyticus species, the multiplex PCR developed in this study will be very effective in epidemiological, ecological and economical points of view.
Assuntos
Proteínas de Bactérias/genética , Técnicas de Tipagem Bacteriana/métodos , Proteínas Hemolisinas/genética , Reação em Cadeia da Polimerase/métodos , Vibrio parahaemolyticus/isolamento & purificação , Animais , Penaeidae , Reação em Cadeia da Polimerase/economia , Sensibilidade e Especificidade , Vibrio/genética , Vibrio/isolamento & purificação , Vibrioses/diagnóstico , Vibrioses/microbiologia , Vibrio parahaemolyticus/genéticaRESUMO
An experiment was conducted to exploit the immune-modulation effect of trivalent chromium (Cr) for augmenting immunity against peste des petits ruminants (PPR) in dwarf black Bengal goats (Capra hircus). The goats (n = 6 per treatment group) either received a basal diet devoid of supplemental Cr (control) or were supplemented with 0.5 mg Cr as chromic chloride (+Cr) for 60 days. Live weight gain and feed conversion efficiency improved (P < 0.05) in the +Cr dietary group. Supplemental Cr did not affect the total number of monocytes, eosinophils and basophils (P > 0.1) although total leukocytes increased (P < 0.05) and the ratio of neutrophils to lymphocytes narrowed down (P < 0.05) in the +Cr dietary group. The effect of Cr supplementation on variables of the intravenous glucose tolerance test (IVGTT) was inconclusive. Compared to the control group, the basal concentration of glucose (P < 0.05) and insulin (P < 0.001) was higher, clearance rate was slower (P < 0.05) and serum half-life was greater (P < 0.05) in the +Cr dietary group during the IVGTT. Over the duration of the experiment, serum concentration of insulin increased (P < 0.001) and that of cortisol decreased (P < 0.01) in the +Cr group, which also showed a relatively higher primary antibody (Ab) response against PPR on days 10 (P < 0.01) and 20 (P < 0.05) post-vaccination. In conclusion, the experiment indicated that supplementation of Cr as CrCl3, in the diet of non-stressed goats, may improve primary Ab response against PPR and help confer an augmented immunity to the disease besides promoting growth and feed conversion.