Milk samples (675 samples from Alborz province and 260 samples from Tehran province) of all dairy cattle were cultured on Brucella-selective supplement media containing nalidixic acid (2.5 mg), polymyxin B sulfate (2,500 IU), vancomycin (10 mg), nystatin (50,000 IU), cycloheximide (50 mg), and bacitracin (12,500 IU) (HiMedia, Mumbai, India). They were then maintained at 10% CO2 at 37 °C for 14 days [21 ].
Cycloheximide (chx)
Manufactured by HiMedia
Sourced in India
Cycloheximide is a laboratory reagent used primarily as a protein synthesis inhibitor in eukaryotic cells. It acts by blocking the translocation step in protein synthesis, thereby preventing the formation of peptide bonds. Cycloheximide is commonly used in various cell biology and biochemical applications to study cellular processes involving protein synthesis.
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Lab products found in correlation
3 protocols using cycloheximide (chx)
1
Brucella Detection in Dairy Cattle Milk
2
Dermatophyte Isolation and Identification
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Collected samples were cultured on dermatophyte test media (DTM, HiMedia, India, REF: M188-500G) agar plates containing papaic digest of soybean meal (10 g/L), glucose (10 g/L), phenol red (0.2 g/L), agar (20 g/L) with yeast extract (4 g/L), and gentamicin (50 mg/L) with cycloheximide (500 mg/L) (HiMedia Company, India) at a final pH of 5.6 ± 0.2 prepared per the manufacturer’s instructions. All inoculated plates were then incubated at 30°C for 4 weeks. Plates were examined twice a week for any fungal growth, and in the absence of growth during week 4, these samples were considered negative.
Cultures of the isolated dermatophytes were initially identified by examining their colony morphologies (macroscopically) on DTM and microscopic characteristics. Dermatophytes were identified based on the change in DTM color from yellow to red as dermatophytes produce alkali substances that promote the pH and change the medium’s phenol red from yellow to red. For the macroscopic identification, the growth texture, rate, and pigmentation of the reverse and front sides of the culture were employed. Microscopically, the isolates were identified using adhesive tape and lactophenol cotton blue stain (LPCB). The sticky piece of tape with fungal structures adhered to the slide with a layer of LPCB.
Cultures of the isolated dermatophytes were initially identified by examining their colony morphologies (macroscopically) on DTM and microscopic characteristics. Dermatophytes were identified based on the change in DTM color from yellow to red as dermatophytes produce alkali substances that promote the pH and change the medium’s phenol red from yellow to red. For the macroscopic identification, the growth texture, rate, and pigmentation of the reverse and front sides of the culture were employed. Microscopically, the isolates were identified using adhesive tape and lactophenol cotton blue stain (LPCB). The sticky piece of tape with fungal structures adhered to the slide with a layer of LPCB.
3
Identification and Antifungal Susceptibility of Dermatophytes
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All the 395 scraping samples were collected, and the specimens were shipped to a central facility. The primary identification of dermatophytes was done using direct microscopy with 10% potassium hydroxide (KOH) mount. Direct microscopic examination of the wet-mount was performed under a microscope, under ×10 and ×40 for fungal hyphae, spores or yeast cells.
The Sabouraud dextrose agar (SDA) was used for isolation and identification of fungal isolates. Specimens were cultured on SDA media (MicroMaster Laboratories Pvt. Ltd) with 0.05% chloramphenicol alone (MicroMaster Laboratories Pvt. Ltd), or with 0.5% cycloheximide (HiMedia Laboratories Pvt. Ltd) and 0.05% chloramphenicol (MicroMaster Laboratories Pvt. Ltd) and incubated at 30°C for up to four weeks. Cultures were examined once a week and professed negative if no growth was observed until 6 weeks. Identification of dermatophytes to the species level was done by assessing the colony morphology, microscopy (Lactophenol Cotton Blue Mount), and physiological and biochemical tests. Further antifungal drug susceptibility testing was performed, and the minimum inhibitory concentration (MIC) of the drugs was determined.
The Sabouraud dextrose agar (SDA) was used for isolation and identification of fungal isolates. Specimens were cultured on SDA media (MicroMaster Laboratories Pvt. Ltd) with 0.05% chloramphenicol alone (MicroMaster Laboratories Pvt. Ltd), or with 0.5% cycloheximide (HiMedia Laboratories Pvt. Ltd) and 0.05% chloramphenicol (MicroMaster Laboratories Pvt. Ltd) and incubated at 30°C for up to four weeks. Cultures were examined once a week and professed negative if no growth was observed until 6 weeks. Identification of dermatophytes to the species level was done by assessing the colony morphology, microscopy (Lactophenol Cotton Blue Mount), and physiological and biochemical tests. Further antifungal drug susceptibility testing was performed, and the minimum inhibitory concentration (MIC) of the drugs was determined.
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