Comparative study of five culture media for in vitro culture of Trichomonas gallinae from birds in Assam
At a Glance
Section titled âAt a Glanceâ| Metadata | Details |
|---|---|
| Publication Date | 2022-11-01 |
| Journal | Journal of Entomology and Zoology Studies |
| Authors | M Saikia, K Bhattacharjee, Sarmah PC, Deka DK, RANJEET NEOG |
| Institutions | Assam Agricultural University |
| Analysis | Full AI Review Included |
Technical Documentation & Analysis: Advanced Materials for Bio-Research Platforms
Section titled âTechnical Documentation & Analysis: Advanced Materials for Bio-Research PlatformsâExecutive Summary
Section titled âExecutive SummaryâThis research paper details the comparative efficacy of five culture media for the in vitro cultivation of Trichomonas gallinae. While focused on veterinary parasitology, the methodology highlights critical requirements for high-precision biological research, specifically thermal stability, chemical inertness, and high-resolution optical accessâareas where 6CCVDâs MPCVD diamond excels.
- Core Achievement: Medium 199 demonstrated superior performance, achieving peak growth of 16,000 cells/ml and maintaining high motility for 7 days under controlled conditions.
- Methodology Focus: The study relies on strict environmental control, including incubation at 37 °C, media sterilization at 121 °C (15 lb/in2), and precise cell counting using 400X microscopy.
- Material Science Relevance: The need for stable, inert substrates capable of handling sterilization temperatures and providing high optical clarity (for 400X counting) points directly to the use of CVD diamond in advanced bio-research instrumentation.
- Thermal Management: Maintaining the critical 37 °C incubation temperature requires highly stable thermal platforms, a key application for 6CCVDâs SCD and PCD materials.
- Electrochemical Sensing: The studyâs reliance on pH adjustment (to 6.5) suggests potential future integration of electrochemical sensors, a prime application for Boron-Doped Diamond (BDD) electrodes.
- 6CCVD Value Proposition: We provide SCD and PCD substrates with ultra-low surface roughness (Ra < 1nm) and custom metalization, ideal for next-generation microscopy windows, microfluidic chips, and biosensor arrays required to replicate or extend this type of precision research.
Technical Specifications
Section titled âTechnical SpecificationsâThe following data points extracted from the research highlight the precision required for successful in vitro cultivation, emphasizing the need for high-performance materials in supporting instrumentation.
| Parameter | Value | Unit | Context | Material Science Relevance |
|---|---|---|---|---|
| Incubation Temperature | 37 | °C | Standard incubation for T. gallinae growth. | Requires high thermal conductivity (SCD) for uniform temperature control. |
| Sterilization Temperature | 121 | °C | Autoclaving of Modified Diamondâs media. | Diamond is stable across extreme temperature ranges, unlike polymers. |
| Sterilization Pressure | 15 | lb/in2 | Autoclaving pressure. | Diamond substrates offer superior mechanical robustness. |
| Initial Inoculation Density | 1 x 103 | cell/ml | Standardized starting concentration. | Requires highly accurate optical counting systems (SCD windows). |
| Peak Cell Concentration | 16,000 | cell/ml | Highest growth achieved (Medium 199, 48-72 hours). | Demands high-resolution imaging capability (Ra < 1nm polishing). |
| Microscopy Magnification | 400 | X | Used for counting motile organisms. | Requires optical-grade SCD windows for minimal distortion. |
| Media pH Adjustment | 6.5 | N/A | Adjusted using hydrochloric acid. | BDD is highly resistant to corrosive acids and bases for integrated pH sensing. |
| Surface Roughness Requirement | Ra < 1 | nm | Estimated requirement for high-fidelity 400X imaging. | 6CCVD SCD polishing capability meets this requirement. |
Key Methodologies
Section titled âKey MethodologiesâThe experimental procedure emphasizes strict control over chemical composition, temperature, and optical assessment, demonstrating the need for robust, high-purity laboratory components.
- Media Preparation and Sterilization: Modified Diamondâs media components (trypticase, yeast extract, maltose, L-cysteine hydrochloride, ascorbic acid) were dissolved in distilled water and autoclaved at 121 °C under 15 lb/in2 pressure for 15 minutes.
- Supplementation and pH Control: Media was supplemented with an antibiotic mixture (Penicillin G, Streptomycin sulfate) and 10% fresh inactivated fetal calf serum. The pH was critically adjusted to 6.5 using hydrochloric acid.
- Inoculation: Oral swabs were used to inoculate 5 ml culture tubes, standardizing the initial concentration to 1x103 trophozoites/ml.
- Incubation: Culture tubes were incubated in an aerobic environment within a BOD incubator maintained precisely at 37 °C for 7 days.
- Assessment and Counting: Growth and motility were monitored every 24 hours. Live/dead differentiation was achieved using 0.3% Trypan blue dye, followed by counting using a Neubauer chamber at 400X magnification.
- Subculture: Positive culture materials were transferred into fresh media every 48-72 hours to control bacterial contamination.
6CCVD Solutions & Capabilities
Section titled â6CCVD Solutions & CapabilitiesâThis research, while focused on protozoan culture, underscores the need for materials that guarantee precision, chemical inertness, and thermal stability in advanced biological and chemical analysis systems. 6CCVD provides the necessary MPCVD diamond solutions to elevate the performance and reliability of instrumentation used in similar high-stakes laboratory environments.
Applicable Materials
Section titled âApplicable MaterialsâTo replicate or extend this research into integrated microfluidic or sensing platforms, 6CCVD recommends the following materials:
| 6CCVD Material | Application in Bio-Research | Key Benefit |
|---|---|---|
| Optical Grade Single Crystal Diamond (SCD) | High-resolution microscopy windows, UV/IR spectroscopy ports, microfluidic channel covers. | Ultra-low surface roughness (Ra < 1nm) essential for 400X imaging; high transparency across UV-IR spectrum; chemical inertness against media (pH 6.5) and sterilization agents. |
| Polycrystalline Diamond (PCD) | High-power thermal spreaders for incubator heating elements, robust substrates for large-area sensor arrays (up to 125mm). | Excellent thermal conductivity (up to 2000 W/mK) ensures uniform 37 °C incubation temperature across large platforms. |
| Boron-Doped Diamond (BDD) | Integrated electrochemical sensors for real-time monitoring of pH, dissolved oxygen, or metabolic byproducts in culture media. | Wide electrochemical window, extreme chemical stability, and resistance to fouling, ideal for monitoring the 7-day culture cycle. |
Customization Potential
Section titled âCustomization PotentialâThe requirement for precise, sterile, and optically accessible culture environments necessitates custom component design. 6CCVDâs capabilities directly address these needs:
- Custom Dimensions: We supply plates and wafers up to 125mm (PCD) and custom-cut SCD pieces, allowing for the fabrication of unique microfluidic chips or multi-well plates with integrated diamond components.
- Precision Polishing: Our internal capability achieves Ra < 1nm for SCD and Ra < 5nm for inch-size PCD, ensuring optimal optical performance for high-magnification microscopy (400X) and minimal cell adhesion.
- Integrated Metalization: For BDD electrodes or thermal contacts required for precise 37 °C control, 6CCVD offers custom metalization stacks including Ti, Pt, Au, Pd, W, and Cu. This is crucial for creating robust, sterilizable electrical contacts.
- Substrate Thickness: We provide SCD and PCD in thicknesses ranging from 0.1”m (for thin optical membranes) up to 500”m, and robust substrates up to 10mm, ensuring mechanical stability under sterilization pressures (15 lb/in2).
Engineering Support
Section titled âEngineering Supportâ6CCVDâs in-house PhD team specializes in applying diamondâs unique properties to challenging scientific applications. We can assist researchers working on similar Advanced Bio-Sensing and Microfluidic Culture projects by:
- Consulting on optimal diamond grade selection (SCD vs. PCD vs. BDD) based on required thermal, optical, and electrochemical performance.
- Designing custom metalization layouts for integrated heating elements or sensor arrays compatible with high-temperature sterilization protocols.
- Providing DDU and DDP global shipping options to ensure rapid delivery of critical materials worldwide.
For custom specifications or material consultation, visit 6ccvd.com or contact our engineering team directly.
View Original Abstract
A study was carried out in order to compare the in vitro efficacy of five different culture media against the flagellate protozoa, Trichomonas gallinae. For in vitro culture, oral swabs of T. gallinae were collected randomly from oropharynx and crop after visual inspection of birds as pigeon, duck, chicken and quail with a sterilized cotton swab and cultivated in five media namely modified Diamond media, Medium199, Minimum Essential Medium (MEM), RPMI 1640 and Nutrient broth. The initial inoculation of trophozoites into each media was 1X103cell/ml. Among all the five media, Medium 199 showed the highest growth rate and motility of the organisms till 7 days after initial inoculation. All media were found suitable for harvesting and sub culture of T. gallinae under in vitro condition.