Isı Önlenmesi ile Sıçanlarda Olumsuz Etkileri Stres<em> Bacillus subtilis</em> Gerilme
Summary
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.
Abstract
Bu çalışma, sıcak stresine bağlı komplikasyonlar karşı Bacillus subtilis suşu koruyucu etkisini değerlendirmek için tasarlanmıştır. Otuz iki Sprague-Dawley fareleri, bu çalışmada kullanılmıştır. Hayvanlar oral B ile iki gün süreyle günde iki kez uygulandı subtilis BSB3 zorlanma veya PBS. Ertesi gün, son tedaviden sonra, her bir grup bölündü ve iki deney grubu (bir PBS ile muamele edilmiş ve bir B. subtilis ile muamele) 25 dakika süreyle 45 ° C de yerleştirildi. İki kontrol grubu, oda sıcaklığında 25 dakika süreyle kaldı. Tüm sıçanlara ötenazi yapıldı ve farklı parametrelerin tüm gruplarda analiz edildi. Isı stresinin olumsuz etkileri villus yüksekliği ve bağırsak epiteli toplam mukozal kalınlığının azalması ile kayıtlı; lümenden bakteri translokasyonu; Kandaki (LPS) seviyesi lipopolisakkaridinin ait eritrositlerin vezikülasyon ve yükseklik arttı. tedavinin koruyucu etkinliği ön tarafından değerlendirilirBu yan etkilerin giriflim. Protokol ısı stresi komplikasyonların önlenmesi için bakterilerle sıçanların oral tedavi için ayarlanmış, ancak bu protokol değiştirilebilir ve diğer uygulama yolları için ve farklı bileşiklerin analizi için kullanılabilir.
Introduction
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.
Protocol
Representative Results
Discussion
Ciddi sağlık koşullarında 29 yüksek sıcaklık sonuçlarına maruz kalmak. Önleme ve ısı stresi ile ilgili komplikasyonların erken teşhis hayati önem 30 taşımaktadır. sunulan protokol ısı stresi olumsuz etkilerin önlenmesi için çeşitli yaklaşımların etkinliğini değerlendirmek için kullanılabilir.
Bu protokol kapsamında kritik adımlar şunlardır: ısıl işlem prosedürü (45 o C, 25 dakika); toplama ve enteroto…
Disclosures
The authors have nothing to disclose.
Acknowledgements
This work was supported by the Auburn University fund FOP 101002-139294-2050.
Materials
| Phosphate buffered saline (PBS) | Sigma-Aldrich, St. Louis, MO | P4417 | |
| Ethyl Alcohol | Pharmco products Inc. Brookfield, CT, USA | 64-17-5 | |
| Agar | VWR | 97064-334 | |
| MacConkey agar plates | VWR | 470180-742 | |
| 5% blood agar plates | VWR | 89405-024 | |
| Brucella blood with Hemin, and Vitamin K, plates |
VWR | 89405-032 | |
| Bouin’s Fixative | Electron Microscopy Sciences, Hatfield, PA, USA | 15990 | |
| IL-10 Rat ELISA kit | Invitrogen, Camarillo, CA, USA | KRC0101 | |
| IL-1beta Rat ELISA Kit | Invitrogen, Camarillo, CA, USA | KRC0011 | |
| IL-6 Rat ELISA Kit | Invitrogen, Camarillo, CA, USA | KRC0061 | |
| INF-gamma Rat ELISA Kit | Invitrogen, Camarillo, CA, USA | KRC4021 | |
| TNF-alpha Rat ELISA Kit | Invitrogen, Camarillo, CA, USA | KRC3011 | |
| Rat LPS ELISA Kit | NeoBioLab, MA, USA | RL0275 | |
| Environmental Chamber 6020-1 | Caron, Marietta, OH, USA | 6020-1 | |
| Centrifuge | Beckman Coulter, Indianapolis, IN, USA | Optima L-90K | |
| Ultra Centrifuge | |||
| Light microscope optical system | CitoViva Technology Inc., Auburn, AL | ||
| Colony counter | Fisher Scientific , Pittsburgh, PA, USA | RE-3325 | |
| Freezer | Haier, Brooklyn, NY, USA | HCMO50LA | |
| Ultra microplate reader | Bio-Tek Instrument, Winooski, VT, USA | ELx 808 | |
| Auto Strip Washer | Bio-Tek Instrument, Winooski, VT, USA | ELx50 | |
| Pipettes | Gilson, Pipetman, France | P100, P200, P1000 | |
| C24 Incubator Shaker | New Brunswick Scientific, Enfield, CT, USA | Classic C24 | |
| Rocking Shaker | Reliable Scientific, Inc., Nesbit, MS, USA | 55 | |
| Petri dishes | Fisher Scientific, Pittsburgh, PA, USA | 875713 | 100mmX15mm |
| SterilGard III Advance | The Baker Company, Sanford, ME, USA | SG403 | |
| Culture Growing Flasks | Corning Incorporated, Corning, NY, USA | 4995 | PYREX 250mL Erlenmeyer flasks |
| Optical Spectrometer Genesys 20 | Thermo Scientific, Waltham, MA, USA. | 4001 | |
| Sony DXC-33 Video camera | Sony | ||
| Richardson test slide | Electron Microscopy Science, Hatfield, PA, USA | 80303 | |
| Millipore water purification system | Millipore | Direct-Q | |
| Image-Pro Plus software | Media Cybernetics, MD, USA | ||
| Triple Beam Balance | OHAUS Corporation, Parsippany, NJ, USA | ||
| Tissue Homogenizer, Dounce | VWR | 71000-518 | |
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