Bacterial strains having the ability to inhibit the growth of other bacteria were isolated from soil samples collected from Kashmir Smast (smast is Pushto for cave), Khyber Pakhtunkhwa, Pakistan. The study includes mineralogical and geochemical analyses of soil sample collected from the cave, so as to describe the habitat from which the microorganisms have been isolated. Total bacterial count of the soil sample was 5.25 3 10 4 CFU mL −1. Four bacterial isolates having activity against test organisms Micrococcus luteus, Klebsiella sp., Pseudomonas sp., and Staphylococcus aureus were screened out for further study. Two of the isolates were found to be Gram-positive and the other two Gram-negative. The four isolates showing antibacterial activity were identified as Serratia sp. KC1-MRL, Bacillus licheniformis KC2-MRL, Bacillus sp. KC3-MRL, and Stenotrophomonas sp. KC4-MRL on the basis of 16S rRNA sequence analysis. Although all isolates showed antibacterial activity, only Bacillus licheniformis KC2-MRL was selected for further study due to its large zone of inhibition. Antibacterial activity of B. licheniformis KC2-MRL was optimum when grown in nutrient broth adjusted to pH 5 and after 24 hours of incubation at 35 uC. The extracted antibacterial compound was stable at pH 5-7 and 40 uC when incubated for 1 hour. The strain was found resistant against cefotaxime (ctx). Atomic-absorption analysis of the soil sample collected from the cave showed high concentrations of calcium (332.938 mg kg ) compared to the control soil collected outside the cave. FTIR spectrum of the concentrated protein showed similarity to bacitracin. The antibacterial compound showed activity against both Gram-negative and positive test strains. Mineralogy of Kashmir Smast is diverse and noteworthy. Different geochemical classes identified by X-ray diffraction were nitrates, oxides, phosphates, silicates, and sulfates. Weathered cave limestone contributes notably to the formation of these minerals or compounds. FTIR spectroscopic analysis helped to identify minerals such as quartz, clinochlore, vermiculite, illite, calcite, and biotite.

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Abstract. Murree and Ayubia hills are parts of the lower Himalaya Mountain range in Pakistan. The rocks of these hills are younger and mostly limestone in nature. In this study, geomicrobiological investigation of the selected samples collected from Murree and Ayubia hills was carried out. The rock samples were spread on nutrient agar plates and endolithic and epilithic bacteria were isolated on the basis of colony morphology and studied further. All the isolates were screened for different industrially important hydrolytic enzymes. Study isolates were identified through 16S rRNA gene sequencing. These endoliths and epiliths were assayed for antimicrobial activities against clinical isolates S. aureus, P. aeuroginosa and E. coli. Total 31 bacterial isolates were recovered of which 15 were endolithic and 16 were epilithic. 16S rRNA gene sequencing revealed major culturable groups colonizing these rocks were clustered in four major groups, Proteobacteria, Firmicutes, Actinobacteria and Bacteroidetes which inlcude Alcaligenes, Lysinibacillus, Actinobacteria, Pseudomonas, Pusillimonas, Streptomyces, Fluviicola, Serratia, Flavobacterium, Stenotrophomonas and Brevindomonas species. The sequences were deposited in NCBI for acquisition of accession numbers. The bacterial isolates were efficient producers of oxidase, catalase, protease, amylase and gelatinase. The endolithic isolates N4 (Pseudomonas sp. KT223616) and N28 (Streptomyces sp. KT004386) exhibited good activity against all the three clinically isolated target strains. The study revealed the rocks of Murree and Ayubia hills have a rich microbial ecology that besides having an important role in weathering and mineralization processes may also be potential source of biotechnological applications.

The Karakoram Mountain Range (KMR) is one of the largest mountain ranges in the world, with ~ 37% of its area glaciated. Here, we present the geochemistry of ice, sediment and meltwaters sampled from Ghulmet, Ghulkin and Hopar glaciers of the Karakoram Range, Pakistan, in addition to the first information on the diversity of pigmented bacteria evaluated using culture-dependent techniques. Geochemical analyses revealed Ca2+ and SO42− to be the most abundant cation and anion species across all glacial samples, respectively. Total organic carbon (TOC), total nitrogen (TN) and total phosphorus (TP) were found in the sediments of all glaciers studied in current research. Bacterial species were capable of producing a variety of different pigments, including alloxanthin, astaxanthin, bacterioruberin, β-carotene, 19'-hexanoyloxyfucoxanthin, peridinin, violacein and zeaxanthin. Culturable bacterial diversity was studied using two molecular biomarkers, 16S rRNA and rpoB gene, with a total of 82 bacterial strains representing 25 genera identified across all glacial samples. This study provides the first characterization of glacier-associated, pigment-producing bacterial communities from the KMR. Findings are important for considerations of alternative sources of conventional pigment production in industrial fields.