Growth Curves of Anaerobic Bacteria in Solid Media

Casciato, Dennis A.; Stewart, P. R.; Rosenblatt, Jon E.

doi:10.1128/aem.29.5.610-614.1975

Cited by 6 publications

(4 citation statements)

References 7 publications

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“…Reproducible and efficient data analysis methods are required for analysis of growth measurements from high‐throughput methods. This protocol details the equations required to convert microplate reader absorbance measurements over time to approximate growth rates in colony forming units over time (CFU/hr) using McFarland standards (Casciato, Stewart, & Rosenblatt, 1975; McFarland, 1907). Analysis was conducted in R v. 4.1.2 (R Core Team, 2014), producing figures using the package ggplot2 v. 3.3.2 (Wickham, 2016).…”

Section: Discussionmentioning

confidence: 99%

High‐throughput Method for Novel Medium Development for Culture of Anaerobic Gut Bacteria

et al. 2022

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Gut microbiota play important roles in the health of their host and detailed investigation of these organisms requires in vitro culture. Culturing strictly anaerobic bacteria can be a challenge as the gut environment they inhabit is nutritionally complex. Use of complex media containing nutritionally rich but undefined gut fluid reduces the accuracy of physiological and metabolomic studies. Here we present a high-throughput protocol for comparing growth rates of fastidiously anaerobic bacteria on different media. These protocols can be used to develop a solid medium made up of commercially sourced ingredients, providing replicable growth conditions for previously uncultured anaerobic bacteria. As many fastidious bacteria grow poorly in a liquid broth, these protocols measure bacterial growth rate on solid media. These protocols speed up and simplify the growth rate measurement process by using a multiwell format and equations in place of physical McFarland standards to calculate approximate cell density. Bacterial strains belonging to the families Erysipelotrichaceae and Lachnospiraceae (phylum Firmicutes) isolated from the hindgut of Kyphosus sydneyanus were used to demonstrate the efficacy of these protocols. Bacterial growth rates were compared between a nutritionally rich medium with gut fluid versus a novel replicable medium with mannitol.

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Section: Discussionmentioning

confidence: 99%

High‐throughput Method for Novel Medium Development for Culture of Anaerobic Gut Bacteria

et al. 2022

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“…Biofilm growth on anode surface also mediates direct electron transfer , . Microbial growth rate was relatively rapid in aerobic oxidation (10−20 min) compared to fermentation (>90 min) , . Biofilm also results in higher substrate degradation due to hyper metabolic activity and tends to liberate more number of protons and electrons as evidenced by the current in CV especially in aerobic operation.…”

Section: Discussionmentioning

confidence: 99%

Effect of Anodic Metabolic Function on Bioelectricity Generation and Substrate Degradation in Single Chambered Microbial Fuel Cell

Mohan

Mohanakrishna

Sarma

2008

Environ. Sci. Technol.

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Influence of anodic metabolic function viz., aerobic, anoxic and anaerobic on bioelectricity generation was evaluated in single chamber mediatorless microbial fuel cells (non-catalyzed graphite electrodes; open-air cathode) during wastewater treatment under similar operating conditions (pH 7; ambient temperature/pressure). Despite the fluctuations observed, aerobic metabolic function (379 mV; 538 mA/m2) documented higher power generation compared to anoxic (251 mV; 348 mA/m2) and anaerobic (265 mV; 211 mA/m2) operations. Relatively higher treatment efficiency was also evidenced in aerobic operation (COD removal efficiency; 77.68% (aerobic), 56.84% (anoxic), 48.68% (anaerobic)). Polarization behavior, bioelectrochemical analysis, sustainable resistance and cell potentials also supported the aerobic operation. Aerobic metabolic function showed potential to generate higher power and substrate degradation over the corresponding anoxic and anaerobic metabolic functions. The relative efficiency of power generation observed in aerobic microenvironment might be attributed to effective substrate oxidation and good biofilm growth observed on the anodic surface. Presence of lower dissolved oxygen concentration in anodic chamber due to the establishment of equilibrium between substrate oxidation and oxygen scavenging might also contributes positively to power generation in aerobic operation.

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“…Optical density (OD) measurements with a spectrophotometer can determine the growth curve of bacteria over time in a liquid sample [ 74 – 78 ]. Initial attempts to measure the bacterial growth by OD were performed in 1974 [ 79 ], but found greater application after the turn of the century. The OD of a bacterial suspension is determined at regular time points upon exposure to an antimicrobial compound, and can, thus, discriminate resistant and sensitive strains (Fig.…”

Section: Optical Methodsmentioning

confidence: 99%

In vitro antimicrobial susceptibility testing methods: agar dilution to 3D tissue-engineered models

Schumacher

Vranken

Malhotra

et al. 2017

Eur J Clin Microbiol Infect Dis

101

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In the field of orthopaedic surgery, bacterial invasion of implants and the resulting periprosthetic infections are a common and unresolved problem. Antimicrobial susceptibility testing methods help to define the optimal treatment and identify antimicrobial resistance. This review discusses proven gold-standard techniques and recently developed models for antimicrobial susceptibility testing, while also providing a future outlook. Conventional, gold-standard methods, such as broth microdilution, are still widely applied in clinical settings. Although recently developed methods based on microfluidics and microdroplets have shown advantages over conventional methods in terms of testing speed, safety and the potential to provide a deeper insight into resistance mechanisms, extensive validation is required to translate this research to clinical practice. Recent optical and mechanical methods are complex and expensive and, therefore, not immediately clinically applicable. Novel osteoblast infection and tissue models best resemble infections in vivo. However, the integration of biomaterials into these models remains challenging and they require a long tissue culture, making their rapid clinical implementation unlikely. A method applicable for both clinical and research environments is difficult to realise. With a continuous increase in antimicrobial resistance, there is an urgent need for methods that analyse recurrent infections to identify the optimal treatment approaches. Graphical abstractTimeline of published and partly applied antimicrobial susceptibility testing methods, listed according to their underlying mechanism, complexity and application in research or clinics.

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Growth Curves of Anaerobic Bacteria in Solid Media

Cited by 6 publications

References 7 publications

High‐throughput Method for Novel Medium Development for Culture of Anaerobic Gut Bacteria

High‐throughput Method for Novel Medium Development for Culture of Anaerobic Gut Bacteria

Effect of Anodic Metabolic Function on Bioelectricity Generation and Substrate Degradation in Single Chambered Microbial Fuel Cell

In vitro antimicrobial susceptibility testing methods: agar dilution to 3D tissue-engineered models

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