Dataset Information

Corneal Culture in Infectious Keratitis: Effect of the Inoculation Method and Media on the Corneal Culture Outcome.

ABSTRACT:

Background

To compare two different methods of corneal culture in infectious keratitis: multiple sampling for direct inoculation and enrichment (standard method) and a single sample via transport medium for indirect inoculation (indirect inoculation method).

Methods

Prospective inclusion of patients fulfilling predefined criteria of infectious keratitis undergoing corneal culture according to both studied methods in a randomized order.

Results

The standard method resulted in a significantly higher proportion of positive culture outcomes among the 94 included episodes of infectious keratitis (61%; 57/94) than the indirect inoculation method (44%; 41/94) (p = 0.002) and a significantly higher proportion of microorganisms than the indirect inoculation method, with a Cohen's kappa of 0.38 (95% CI: 0.28-0.49) for agreement between the methods. Subanalysis of culture results showed that direct inoculation on gonococcal agar only combined with the indirect inoculation method resulted in a similar rate of culture positive patients and proportion of detected microorganisms to the standard method.

Conclusion

Indirect inoculation of one corneal sample cannot replace direct inoculation of multiple corneal samples without loss of information. A combination of directly and indirectly inoculated samples can reduce the number of corneal samples by four without statistically significant differences in culture outcome or in the proportion of detected microorganisms.

SUBMITTER: Sagerfors S

PROVIDER: S-EPMC8122416 | biostudies-literature |

REPOSITORIES: biostudies-literature

ACCESS DATA

Similar Datasets

Project description:PurposeTo determine whether convolutional neural networks can detect morphological differences between images of microbiologically positive and negative corneal ulcers.MethodsA cross-sectional comparison of prospectively collected data consisting of bacterial and fungal cultures and smears from eyes with acute infectious keratitis at Aravind Eye Hospital. Two convolutional neural network architectures (DenseNet and MobileNet) were trained using images obtained from handheld cameras collected from culture-positive and negative images and smear-positive and -negative images. Each architecture was trained on two image sets: (1) one with labels assigned using only culture results and (2) one using culture and smear results. The outcome measure was area under the receiver operating characteristic curve for predicting whether an ulcer would be microbiologically positive or negative.ResultsThere were 1970 images from 886 patients were included. None of the models were better than random chance at predicting positive microbiologic results (area under the receiver operating characteristic curve ranged from 0.49 to 0.56; all confidence intervals included 0.5).ConclusionsThese two state-of-the-art deep convolutional neural network architectures could not reliably predict whether a corneal ulcer would be microbiologically positive or negative based on clinical photographs. This absence of detectable morphological differences informs the future development of computer vision models trained to predict the causative agent in infectious keratitis using corneal photography.Translational relevanceThese deep learning models were not able to identify morphological differences between microbiologically positive and negative corneal ulcers. This finding suggests that similar artificial intelligence models trained to identify the causative pathogen using only microbiologically positive cases may have potential to generalize well, including to cases with falsely negative microbiologic testing.

Project description:For the past 150 years, bacteria have been investigated primarily in liquid batch cultures. Contrary to most expectations, these cultures are not homogeneous mixtures of single-cell bacteria, because free-floating bacterial aggregates eventually develop in most liquid batch cultures. These aggregates share characteristics with biofilms, such as increased antibiotic tolerance. We investigated how aggregates develop and what influences this development in liquid batch cultures of Pseudomonas aeruginosa We focused on how the method of inoculation affected aggregation by assessing aggregate frequency and size using confocal laser scanning microscopy. Several traditional methods of initiating an overnight bacterial culture, i.e., inoculation directly from frozen cultures, inoculation using agar-grown cells, or inoculation using cells grown in liquid cultures, were investigated. We discovered a direct link between the inoculation method and the size and frequency of biofilm aggregates in liquid batch cultures, with inoculation directly from a plate resulting in the most numerous and largest aggregates. These large aggregates had an overall impact on the cultures' subsequent tolerance toward tobramycin, indicating that the inoculation method has a profound impact on antibiotic tolerance. We also observed a mechanism whereby preformed aggregates recruited single cells from the surrounding culture in a "snowball effect," building up aggregated biomass in the culture. This recruitment was found to rely heavily on the exopolysaccharide Psl. Additionally, we found that both Escherichia coli and Staphylococcus aureus produced aggregates in liquid batch cultures. Our results stress the importance of inoculation consistency throughout experiments and the substantial impact aggregate development in liquid batch cultures may have on the outcomes of microbiological experiments.IMPORTANCE Pure liquid cultures are fundamental to the field of microbiological research. These cultures are normally thought of as homogeneous mixtures of single-cell bacteria; the present study shows that this is not always true. Bacteria may aggregate in these liquid cultures. The aggregation can be induced by the method chosen for inoculation. The presence of aggregates can significantly change the outcomes of experiments by altering the phenotype of the cultures. The study found a mechanism whereby preformed aggregates are able to recruit surrounding single cells in a form of snowball effect, creating more and larger aggregates in the cultures. Once formed, these aggregates are hard to remove. Aggregates in liquid cultures may be an immense unseen challenge for microbiologists.