## 4 Discussion  
There have been several attempts over the last few decades to replenish the antibiotic discovery pipeline. This has become more urgent given the rise in reports of antimicrobial resistance [45]. Rather than repeat older methods of antibiotic discovery, researchers have tried new innovations. One such strand is the examination of traditional medicines for antimicrobial producing bacteria or antimicrobials themselves. This follows from successful generation of lead compounds in the field of cancer [9]. \begin{table}
\begin{tabular}{c c} \hline \hline
**Biological Role.** & **Protein Name** \\ \hline Aminoglycosoxide adenylyltransferases & Aminoglycosoxide N6’-acetyltransferase (EC 2.3.1.82) \\ Resistance to fluoroquinolones & DNA gyrase subunit B (EC 5.99.1.3) \\ Resistance to fluoroquinolones & DNA gyrase subunit A (EC 5.99.1.3) \\ Beta-lactamase & Beta-lactamase \\ Copper homeostasis & Multicopter oxidase \\ Copper homeostasis & Copper resistance protein \\ Copper homeostasis & Copper-translocating P-type ATPase (EC 3.6.3.4) \\ Copper homeostasis & Copper resistance protein \\ Copper homeostasis: copper tolerance & Multidrug resistance transporter, \\ Magnesium and cobalt tolerance & Cytoplasmic copper homeostasis protein \\ Cobalt–zinc–cadmium resistance & Magnesium and cobalt efflux protein \\ Cobalt–zinc–cadmium resistance & Cobalt–zinc–cadmium resistance protein \\  & Transcriptional regulator \\ Mercuric reductase & FAD-dependent NAD(P)-disulphide oxidoreductase \\ \hline \hline \end{tabular}
\end{table} 
Table 6: Antibiotic and metal resistance genes identified from WGS (whole genome sequences) of _Streptomyces_ sp. CJ13 (predicted by Rapid Annotation using Subsystem Technology (RAST)). \begin{table}
\begin{tabular}{c c c} \hline \hline
**Type** & **Antibiotic Gene Cluster** & **Similarity** \\ \hline NRPS & Salinichelins & 53\% \\ NRPS & Antifungal factor & 50\% \\ T1 PKS & Mediomycin A & 46\% \\ NRPS & Weishannycin & 45\% \\ T1 PKS, NRPS & Combamide (macrolactam) & 44\% \\ Lanthipptide & SAL-2242 & 44\% \\ \hline \hline \end{tabular}
\end{table} 
Table 5: Summary of antiSMASH (antibiotics and secondary metabolite analysis shell) 4.1.0 secondary metabolite prediction of _Streptomyces_ sp. strain CJ13 (values above 40% gene similarity). In this manuscript, we isolated a species of _Streptomyces_ (_Streptomyces_ sp. CJ13) from an alkaline grassland soil traditionally known for its healing properties. This had a broad range of inhibitory activity against multiresistant bacteria, anaerobes and fungi. There are many antibiotic producing species of _Streptomyces_ associated with traditional medicines, both as endophytes and epiphytes [10,11]. Many of the discoveries during the golden age of antibiotics also came from soils. The difference between the older types of antimicrobial surveys in soils and those that have been recently isolated from traditional medicine is their specific soil character and location. For instance, Kisameet clay is based on a glacial clay deposit found in British Columbia that inhibits ESKAPE pathogens [13,14]. Similarly, traditional soil discoveries in Jordan focus on red soil that inhibits various pathogens [12]. Whilst our experiments were based on alkaline grassland soil, we did not test the whole soil for antimicrobial activity but rather tried to selectively isolate _Streptomyces_ from the soil. Other studies have shown that karst regions, i.e. regions where there is dissolution of the rocks, especially limestone, support the growth of many _Streptomyces_[46, 47, 48]. By coincidence, some of the closest phylogenetic matches to _Streptomyces_ sp. CJ13 were _Streptomyces_ sp. ICC 1 and ICC4 originating from the Iron Curtain Cave in Chilliwack, British Columbia [32]. Both the West Fermanagh Scarplands and Iron Curtain Cave are regions of karst and share other botanical species including _Spiranthes romanzoffiana_ (Irish lady's-tresses) and _Sisyrinchium bermudiana_ (blue-eyed grass).