Name: prevention of heat stress adverse effects in rats by bacillus subtilis strain
Uploaded: 2016-07-11T14:00:00
Description: Watch this Scientific Journal Video about Prevention of Heat Stress Adverse Effects in Rats by Bacillus subtilis Strain at JoVE.com

This work was supported by the Auburn University fund FOP 101002-139294-2050.

Todos los procedimientos experimentales fueron aprobados por el IACUC de la Universidad de Auburn, Auburn, Alabama. 1. Preparación de medios de cultivo, platos y Cultura bacteriana <ol><li> Inocular 10 ml de caldo nutritivo en un matraz con una sola colonia de la cultura BSB3 Bacillus subtilis, que se cultiva durante la noche en una placa de agar nutriente. </li><li> Hacer 500 ml de medio de esporulación 19 (por litro: Bacto Nutrient Broth, 8 g; 10% (w / v) de KCl, 10 ml; 1,2% (w / v) <su...

<ol>
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Rev. 32, 185-190 (2004).</li><li>Yu, J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effect+of+heat+stress+on+the+porcine+small+intestine:+A+morphological+and+gene+expression+study.">Effect of heat stress on the porcine small intestine: A morphological and gene expression study.</a> Comp. Biochem. Physiol. A-Mol. Integr. Physiol. 156, 119-128 (2010).</li><li>Moseley, P. L., Gisolfi, C. V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=New+Frontiers+in+Thermoregulation+and+Exercise.">New Frontiers in Thermoregulation and Exercise.</a> Sports Med. 16, 163-167 (1993).</li><li>Lambert, G. 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W., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Microbes,+intestinal+inflammation+and+probiotics.">Microbes, intestinal inflammation and probiotics.</a> Expert Rev Gastroent. 6, 81-94 (2012).</li><li>Quigley, E. M. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Prebiotics+and+Probiotics:+Their+Role+in+the+Management+of+Gastrointestinal+Disorders+in+Adults.">Prebiotics and Probiotics: Their Role in the Management of Gastrointestinal Disorders in Adults.</a> Nutr Clin Pract. 27, 195-200 (2012).</li><li>Gore, C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Treatment+and+secondary+prevention+effects+of+the+probiotics+Lactobacillus+paracasei+or+Bifidobacterium+lactis+on+early+infant+eczema:+randomized+controlled+trial+with+follow-up+until+age+3+years.">Treatment and secondary prevention effects of the probiotics Lactobacillus paracasei or Bifidobacterium lactis on early infant eczema: randomized controlled trial with follow-up until age 3 years.</a> Clin Exp Allergy. 42, 112-122 (2012).</li><li>Gomes, A. C., Bueno, A. A., de Souza, R. G. M., Mota, J. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Gut+microbiota,+probiotics+and+diabetes.">Gut microbiota, probiotics and diabetes.</a> Nutr. J. 13, (2014).</li><li>Eutamene, H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Synergy+between+Lactobacillus+paracasei+and+its+bacterial+products+to+counteract+stress-induced+gut+permeability+and+sensitivity+increase+in+rats.">Synergy between Lactobacillus paracasei and its bacterial products to counteract stress-induced gut permeability and sensitivity increase in rats.</a> J. 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A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Time+course+of+cytokine,+corticosterone,+and+tissue+injury+responses+in+mice+during+heat+strain+recovery.">Time course of cytokine, corticosterone, and tissue injury responses in mice during heat strain recovery.</a> J. Appl. Physiol. 100, 1400-1409 (2006).</li><li>Ribeiro, S. R., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Weight+loss+and+morphometric+study+of+intestinal+mucosa+in+rats+after+massive+intestinal+resection.+Influence+of+a+glutamine-enriched+diet.">Weight loss and morphometric study of intestinal mucosa in rats after massive intestinal resection. Influence of a glutamine-enriched diet.</a> Rev. Hosp. Cl&#236;n. Fac. Med. S. 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La exposición a altas temperaturas en condiciones graves de salud 29. La prevención y la detección precoz de complicaciones relacionadas con el estrés por calor son de vital importancia 30. El protocolo presentado se puede utilizar para evaluar la eficacia de los diversos enfoques para la prevención del estrés por calor efectos adversos. Los pasos críticos dentro de este protocolo son: el procedimiento de tratamiento térmico (45 ° ...

Different stressors affect human and animal health. Temperature is one of the most stressful factors, causing chronic, acute and even lethal illnesses1. Changes in intestinal morphology and a loss of gut barrier integrity after heat stress were documented in many cases2,3. This protective barrier is responsible for defense against translocation of gut bacteria and their toxins, in particular lipopolysaccharides (LPS), from the lumen to the internal circulation4,5. Stability of the gut microbiota significantly influences intestinal barrier function, the immune response of the host, and tolerance to stress conditions6. Thus, modulation of the intestinal microbiota can provide a novel approach for prevention of stress-related adverse effects. 
Beneficial bacteria have been used as a promising strategy to modify the gut microbiota for successful management of various clinical conditions, such as diarrhea, inflammatory bowel disease, atopic dermatitis, metabolic disorders7-10. Probiotic effects of beneficial bacteria include production of essential metabolites to support intestinal health, stimulation of the immune system, promotion of lactose tolerance, restoration of epithelial dysfunction. Probiotics were effective in prevention of the stress-related complications in vitro and in animal models11,12. 
Researchers have given more attention to Bacillus bacteria as probiotics because these bacteria support intestinal homeostasis13 and have beneficial effects on the host14. Our previous data demonstrated high efficacy of Bacillus probiotics against pathogens in vitro15,16 and in clinical trials17,18. Here, we aim to evaluate the preventive effect of B. subtilis BSB3 strain against complications after heat stress.

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			<td rowspan="2">Phosphate buffered saline (PBS)</td>
			<td>Sigma-Aldrich,&#160;St. Louis, MO</td>
			<td rowspan="2">P4417</td>
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			<td></td>
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			<td>Ethyl Alcohol&#160;</td>
			<td>Pharmco products Inc. Brookfield, CT, USA&#160;</td>
			<td>64-17-5</td>
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			<td>Agar</td>
			<td>VWR</td>
			<td>97064-334</td>
			<td></td>
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			<td>MacConkey agar plates</td>
			<td>VWR</td>
			<td>470180-742</td>
			<td></td>
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		<tr>
			<td>5% blood agar plates</td>
			<td>VWR</td>
			<td>89405-024</td>
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			<td>Brucella blood with Hemin, and Vitamin K, 
			&#160;plates</td>
			<td>VWR</td>
			<td>89405-032</td>
			<td></td>
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			<td>Bouin&#8217;s Fixative</td>
			<td>Electron Microscopy Sciences, Hatfield, PA, USA</td>
			<td>15990</td>
			<td></td>
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		<tr>
			<td>IL-10 Rat ELISA kit</td>
			<td>Invitrogen, Camarillo, CA, USA</td>
			<td>KRC0101</td>
			<td></td>
		</tr>
		<tr>
			<td>IL-1beta Rat ELISA Kit</td>
			<td>Invitrogen, Camarillo, CA, USA</td>
			<td>KRC0011</td>
			<td></td>
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			<td>IL-6 Rat ELISA Kit</td>
			<td>Invitrogen, Camarillo, CA, USA</td>
			<td>KRC0061</td>
			<td></td>
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			<td>INF-gamma Rat ELISA Kit</td>
			<td>Invitrogen, Camarillo, CA, USA</td>
			<td>KRC4021</td>
			<td></td>
		</tr>
		<tr>
			<td>TNF-alpha Rat ELISA Kit</td>
			<td>Invitrogen, Camarillo, CA, USA</td>
			<td>KRC3011</td>
			<td></td>
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		<tr>
			<td>Rat LPS ELISA Kit</td>
			<td>NeoBioLab, MA, USA</td>
			<td>RL0275</td>
			<td></td>
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		<tr>
			<td>Environmental Chamber 6020-1</td>
			<td>Caron, Marietta, OH, USA</td>
			<td>6020-1</td>
			<td></td>
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			<td rowspan="2">Centrifuge&#160;</td>
			<td rowspan="2">Beckman Coulter, Indianapolis, IN, USA</td>
			<td>Optima L-90K</td>
			<td rowspan="2"></td>
		</tr>
		<tr>
			<td>Ultra Centrifuge</td>
		</tr>
		<tr>
			<td>Light microscope optical system</td>
			<td>CitoViva Technology Inc., Auburn, AL</td>
			<td></td>
			<td></td>
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		<tr>
			<td>Colony counter</td>
			<td>Fisher Scientific , Pittsburgh, PA, USA</td>
			<td>RE-3325</td>
			<td></td>
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			<td>Freezer</td>
			<td>Haier, Brooklyn, NY, USA</td>
			<td>HCMO50LA</td>
			<td></td>
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		<tr>
			<td>Ultra microplate reader</td>
			<td>Bio-Tek Instrument, Winooski, VT, USA</td>
			<td>ELx 808</td>
			<td></td>
		</tr>
		<tr>
			<td>Auto Strip Washer</td>
			<td>Bio-Tek Instrument, Winooski, VT, USA</td>
			<td>ELx50</td>
			<td></td>
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			<td>Pipettes</td>
			<td>Gilson, Pipetman, France</td>
			<td>P100, P200, P1000</td>
			<td></td>
		</tr>
		<tr>
			<td>C24 Incubator Shaker</td>
			<td>New Brunswick Scientific, Enfield, CT, USA</td>
			<td>Classic C24</td>
			<td></td>
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		<tr>
			<td>Rocking Shaker&#160;</td>
			<td>Reliable Scientific, Inc., Nesbit, MS, USA</td>
			<td>55</td>
			<td></td>
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		<tr>
			<td>Petri dishes</td>
			<td>Fisher Scientific, Pittsburgh, PA, USA</td>
			<td>875713</td>
			<td>100mmX15mm</td>
		</tr>
		<tr>
			<td>SterilGard III Advance</td>
			<td>The Baker Company, Sanford, ME, USA</td>
			<td>SG403</td>
			<td></td>
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			<td>Culture Growing Flasks</td>
			<td>Corning Incorporated, Corning, NY, USA</td>
			<td>4995</td>
			<td>PYREX 250mL Erlenmeyer flasks</td>
		</tr>
		<tr>
			<td>Optical Spectrometer Genesys 20</td>
			<td>Thermo Scientific, Waltham, MA, USA.</td>
			<td>4001</td>
			<td></td>
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			<td>Sony DXC-33 Video camera</td>
			<td>Sony</td>
			<td></td>
			<td></td>
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			<td>Richardson test slide</td>
			<td>Electron Microscopy Science, Hatfield, PA, USA</td>
			<td>80303</td>
			<td></td>
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			<td>Millipore water purification system</td>
			<td>Millipore</td>
			<td>Direct-Q</td>
			<td></td>
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			<td>Image-Pro Plus software</td>
			<td>Media Cybernetics, MD, USA</td>
			<td></td>
			<td></td>
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		<tr>
			<td>Triple Beam Balance</td>
			<td>OHAUS Corporation, Parsippany, NJ, USA</td>
			<td></td>
			<td></td>
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			<td rowspan="3">Tissue Homogenizer, Dounce</td>
			<td rowspan="3">VWR</td>
			<td rowspan="3">71000-518</td>
			<td rowspan="3"></td>
		</tr>
		<tr>
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prevention of heat stress adverse effects in rats by bacillus subtilis strain

Este estudio fue diseñado para evaluar el efecto protector de la cepa de Bacillus subtilis contra las complicaciones relacionadas con el estrés por calor. Treinta y dos ratas Sprague-Dawley se utilizaron en este estudio. Los animales se trataron por vía oral dos veces al día durante dos días con B. subtilis cepa BSB3 o PBS. El día siguiente después del último tratamiento, cada grupo se dividió y se colocaron dos grupos experimentales (uno tratado con PBS y uno tratado con B. subtilis) a 45 ° C durante 25 min. Dos grupos de control permanecieron durante 25 min a temperatura ambiente. Todas las ratas se sacrificaron y los diferentes parámetros se analizaron en todos los grupos. Los efectos adversos del estrés por calor son registrados por la disminución de la altura de las vellosidades y el grosor total de la mucosa en el epitelio intestinal; translocación de bacterias desde el lumen; aumento de la vesiculación de los eritrocitos y la elevación de los lipopolisacáridos (LPS) el nivel en la sangre. La eficacia protectora de tratamiento se evaluó por prevención de estos efectos secundarios. El protocolo se creó para el tratamiento oral de ratas con bacterias para la prevención de complicaciones de estrés térmico, pero este protocolo puede ser modificado y utilizado para otras vías de administración y para el análisis de diferentes compuestos.

La temperatura corporal media de los animales antes e inmediatamente después del estrés por calor fue de 36,7 ± 0,07 ° C y 40,3 ± 0,17 ° C, respectivamente (P &lt;0,05). La exposición de ratas al calor (grupo 3) resultó en una inhibición significativa de la altura de las vellosidades y el grosor de la mucosa total de (Figuras 2, 3). La administración oral de la cepa BSB3 antes protegida estrés intestino de los efectos nocivos de calor. </...

Watch this Scientific Journal Video about Prevention of Heat Stress Adverse Effects in Rats by Bacillus subtilis Strain at JoVE.com

Prevention of Heat Stress Adverse Effects in Rats by Bacillus subtilis Strain

We described a protocol for prevention of heat stress effects in rats by oral pre-treatment with beneficial bacteria. This protocol can be modified and used for various routes of administration and for analysis of different compounds.

Prevención de Estrés Térmico efectos adversos en ratas<em&gt; Bacillus subtilis</em&gt; Strain

This study was designed to evaluate the protective effect of the Bacillus subtilis strain against complications related to heat stress. Thirty-two ...

Research

JoVE Journal

Immunology and Infection

Prevention of Heat Stress Adverse Effects in Rats by Bacillus subtilis Strain

Live Cell Imaging of Bacillus subtilis and Streptococcus pneumoniae using Automated Time-lapse Microscopy

Live Cell Imaging of Bacillus subtilis and Streptococcus pneumoniae using Automated Time-lapse Microscopy

Imágenes de células vivas de Bacillus subtilis Y Streptococcus pneumoniae Automatizada utilizando Time-lapse microscopía

During the last few years scientists became increasingly aware that average data obtained from microbial population based experiments are not ...

Investigación

Inmunología e Infección

Introducing Shear Stress in the Study of Bacterial Adhesion

La introducción de la tensión de cizallamiento en el Estudio de la adhesión bacteriana

During bacterial infections a sequence of interactions occur between the pathogen and its host. Bacterial adhesion to the host cell surface is often the ...

Single-cell Analysis of Bacillus subtilis Biofilms Using Fluorescence Microscopy and Flow Cytometry

Single-cell Analysis of Bacillus subtilis Biofilms Using Fluorescence Microscopy and Flow Cytometry

Sola célula de análisis de Bacillus subtilis Las biopelículas mediante microscopía de fluorescencia y citometría de flujo

Biofilm formation is a general attribute to almost all bacteria 1-6. When bacteria form biofilms, cells are encased in extracellular matrix that is mostly ...

Flow Cytometric Analysis of Particle-bound Bet v 1 Allergen in PM10

Análisis de citometría de flujo de partículas unida a Bet v 1 de alérgenos en el MP10

Flow cytometry is a method widely used to quantify suspended solids such as cells or bacteria in a size range from 0.5 to several tens of micrometers in ...

Visualizing the Effects of Sputum on Biofilm Development Using a Chambered Coverglass Model

La visualización de los Efectos de esputo en el desarrollo del biofilm mediante un modelo de Septadas cubreobjetos

Biofilms consist of groups of bacteria encased in a self-secreted matrix. They play an important role in industrial contamination as well as in the ...

Swab Sampling Method for the Detection of Human Norovirus on Surfaces

Método de muestreo hisopo para la Detección de norovirus humano en superficies

Human noroviruses are a leading cause of epidemic and sporadic gastroenteritis worldwide. Because most infections are either spread directly via the ...

Effects of Exposure of Formaldehyde to a Rat Model of Atopic Dermatitis Induced by Neonatal Capsaicin Treatment

Efectos de la exposición de formaldehído a un modelo de rata de la Dermatitis atópica inducida por el tratamiento Neonatal de capsaicina

Atopic dermatitis is chronically relapsing pruritic eczema and prevails around the world especially in developed countries. Complex interactions between ...

Measuring Influenza Neutralizing Antibody Responses to A(H3N2) Viruses in Human Sera by Microneutralization Assays Using MDCK-SIAT1 Cells

Measuring Influenza Neutralizing Antibody Responses to AH3N2 Viruses in Human Sera by Microneutralization Assays Using MDCK-SIAT1 Cells

Medición de Influenza neutralizando las respuestas del anticuerpo al virus a (H3N2) en suero humano por microneutralización ensayos utilizando células de MDCK-SIAT1

Neutralizing antibodies against hemagglutinin (HA) of influenza viruses are considered the main immune mechanism that correlates with protection for ...

Isolation of Salmonella typhimurium-containing Phagosomes from Macrophages

Isolation of Salmonella typhimurium-containing Phagosomes from Macrophages

Aislamiento de Salmonella typhimurium-que contienen fagosomas de los macrófagos

Salmonella typhimurium is a facultative intracellular bacterium that causes gastroenteritis in humans. After invasion of the lamina propria, S. ...

Assessment of the Cytotoxic and Immunomodulatory Effects of Substances in Human Precision-cut Lung Slices

Evaluación de la citotóxica y efectos inmunomoduladores de sustancias en lonchas de pulmón humano de precisión de corte

Respiratory diseases in their broad diversity need appropriate model systems to understand the underlying mechanisms and enable development of new ...