# Metagenomic Analysis of Bacterial Diversity on Reusable Tourniquets in Hospital Environments
## 3 Results  
### Microbial Composition on the Surface of Reusable Tourniquets  
The 16S rRNA sequencing revealed a diverse microbial composition on the surface of reusable tourniquets across different hospital departments. The microbial composition of reusable tourniquets was dominated by Proteobacteria, Firmicutes, Actinobacteria, and the Bacteroidetes family, with _Staphylococcus_, _Pseudomonas_, _Acinetobacter_, and _Enterococcus_  
Figure 2: Heatmap with cluster analysis showing species-level diversity within bacterial genera across tested tourniquets. CFU/cm\({}^{2}\) are sources from the previous article [9]. Based on the division of branches, three distinct clusters (Groups A, B, and C) were identified. These groups represent sets of samples that are more similar to each other within the group than to those in other groups, indicating potential common characteristics or shared patterns. being the most frequently detected genera (Figures 1 and 2). Differences in bacterial composition were observed between emergency departments and operating theatres, with higher bacterial diversity in tourniquets from the operating theatre setting. Figure 2 illustrates the diversity of bacteria in individual tourniquets, based on the number of species occurring within a specific genus. Additionally, data on the colony-forming units per cm\({}^{2}\) of material part of tourniquets, sourced from an earlier publication [9], have been included. High diversity was observed in samples with both low and severe bacterial contamination. For example, samples SBO10_1 (53 CFU/cm\({}^{2}\)) and SBO7_2 (22 CFU/cm\({}^{2}\)) contained, respectively, 33 and 25 genera. Similarly, high bacterial diversity was recorded in a heavily contaminated tourniquet, SBO7_3 (>677 CFU/cm\({}^{2}\)), with 23 genera. Conversely, low species diversity was also observed across both lightly and heavily contaminated samples. For instance, SBO10A_2 (4 CFU/cm\({}^{2}\)) contained only five different genera, and SR6_2 (551 CFU/cm\({}^{2}\)) had just two (_Staphylococcus_ and _Pseudomonas_). There was no correlation between microbiological contamination of the sample (CFU) and species diversity. [PERSON]'s rank correlation analysis showed a value of 0.0897 with \(p\) = 0.7815. Therefore, it can be concluded that there is no clear explanation, and both contamination and species diversity appear to be random. Cluster analysis using the Ward method, based on the number of species occurring within different genera in the samples, allowed for the division of the studied samples into three groups. Group A consists of three samples characterized by the highest diversity of species and genera. These tourniquets originate only from the operating theatre, with an average of 27 different genera per sample. The genera common to all samples in this group include _Pseudomonas_, _Shewanella_, _Acinetobacter_, _Aeromonas_, _Bacillus_, _Serratia_, _Staphylococcus_, _Klebsiella_, _Stenotrophomonas_, _Enterobacter_, and _Enterococcus_. Genera found only in group A are _Shewanella_, _Rahnella_, _Buttiauxella_, _Citrobacter_, _Vibrio_, _Rhizobium_, _Sphygobacterium_, _Timo nella_, _Paracooccus_, _Penaarthobacter_, _Microvigula_, _Lysinibacillus_, _Clostridium_, _Delftia_, _Yersinia_, _Janthinobacterium_, _Pseudoscherichia_, _Silvania_, _Scandinavium_, _Raoultella_, _Providence_, _Proteus_, _Phenylobacterium_, _Pantoea_, _Obesumbacterium_, _Leclercia_, _Hauxiibacter_, _Aeribacillus_, _Cutibacterium_, and _Comomonas_. Group B includes four tourniquets, with half originating from the operating theatre and half from the emergency unit. The average number of genera in this group was 6.5, rounded to 6. The genera common in all samples in this group are _Pseudomonas_, _Acinetobacter_, and _Enterobacter_. Group C consists of five tourniquets, four of which were taken from the emergency unit and one from the operating theatre. The average number of different bacterial genera in this group was 7.2, rounded to 7. The only genus present in all tourniquets within this group was _Staphylococcus_.