Drug resistance and susceptibility testing of Gram negative bacterial isolates from healthy cattle with different β – Lactam resistance Phenotypes from Shandong province China

The objective of this study was to evaluate the effectiveness of common antibiotics against different microorganisms in apparently healthy cattle in Shandong province and its suburb. A total of 220 nasal swab samples were collected and cultured for bacteriological evaluation. All the bacteria isolates after preliminary identification were subjected to antibiogram studies following disc diffusion method. It was found in the study that E. coli is the most commonly associated isolate (21%), followed by Klebsiella spp. (18%), Pseudomonas aeruginosa (13%), Salmonella spp. (15%), Shigella spp (12%), and Proteus spp (11%). While the antibiogram studies reveled that highest number of bacterial isolates showed resistance to Ampicillin (95%), followed by Augmentin (91%), Cefuroxime (85%) and Tetracycline (95%) of (Escherichia coli and Klebsiella spp). In the case of pseudomonas spp. and Salmonella the highest resistance was showed by Ampicillin (90%) followed by Amoxicillin + Clavulanic Acid (80%), Cefixime (90%), and Erythromycin (80%). In Shigella spp and Salmonella spp highest resistance was showed by Amoxicillin, Ceftazidime, Augmentin (60%), and Amoxicillin + Clavulanic Acid (50%). It is concluded that in vitro antibiogram studies of bacterial isolates revealed higher resistance for Ampicillin, Augmentin, Cefuroxime, Cefixime, Tetracycline, Erythromycin, and Amoxicillin + Clavulanic Acid. The high multiple Antibiotics resistance indexes (MARI) observed in all the isolates in this study ranging from 0.6 to 0.9. MARI value of >0.2 is suggests multiple antibiotic resistant bacteria and indicate presence of highly resistant bacteria.

Significant production of vanillin and in vitro amplification of ech gene in local bacterial isolates

Vanillin is the major component which is responsible for flavor and aroma of vanilla extract and is produced by 3 ways: natural extraction from vanilla plant, chemical synthesis and from microbial transformation. Current research was aimed to study bacterial production of vanillin from native natural sources including sewage and soil from industrial areas. The main objective was vanillin bio-production by isolating bacteria from these native sources. Also to adapt methodologies to improve vanillin production by optimized fermentation media and growth conditions. 47 soil and 13 sewage samples were collected from different industrial regions of Lahore, Gujranwala, Faisalabad and Kasur. 67.7% bacterial isolates produced vanillin and 32.3% were non-producers. From these 279 producers, 4 bacterial isolates selected as significant producers were; A3, A4, A7 and A10. These isolates were identified by ribotyping as A3 Pseudomonas fluorescence (KF408302), A4 Enterococcus faecium (KT356807), A7 Alcaligenes faecalis (MW422815) and A10 Bacillus subtilis (KT962919). Vanillin producers were further tested for improved production of vanillin and were grown in different fermentation media under optimized growth conditions for enhanced production of vanillin. The fermentation media (FM) were; clove oil based, rice bran waste (residues oil) based, wheat bran based and modified isoeugenol based. In FM5, FM21, FM22, FM23, FM24, FM30, FM31, FM32, FM34, FM35, FM36, and FM37, the selected 4 bacterial strains produced significant amounts of vanillin. A10 B. subtilis produced maximum amount of vanillin. This strain produced 17.3 g/L vanillin in FM36. Cost of this fermentation medium 36 was 131.5 rupees/L. This fermentation medium was modified isoeugenol based medium with 1% of isoeugenol and 2.5 g/L soybean meal. ech gene was amplified in A3 P. fluorescence using ech specific primers. As vanillin use as flavor has increased tremendously, the bioproduction of vanillin must be focused.

Bacterial Colonization in Computer Keyboards Posses Health Hazard

Computer keyboards of a teaching laboratory were examined and bacteria were isolated from computer keyboards. The subsequent tests were done for the bacterial isolates: methyl red, vogus proskaur, citrate utilization, urease and TSI. This study paves the way to look at an inanimate object like computer keyboard as potential reservoir of bacteria.

Kocuria Strains from Unique Radon Spring Water from Jachymov Spa

Members of the genus Kocuria are often found in soils contaminated with toxic metals or exposed to high levels of ionizing radiation. The use of classical cultivation technics often leads to the isolation of Kocuria sp. from underground spring waters. These bacterial isolates have to adapt their metabolism to survive in such extreme environments. Four bacterial isolates of the genus Kocuria (Kocuria sp. 101, 208, 301, and 401) were obtained from radon spring water (Jachymov, Czech Republic). These isolates were tested for their ability to withstand stress and extreme conditions. Growth was observed at a temperature range of 10–45 °C with optimal growth temperature between 20 and 30 °C. The content of polyunsaturated fatty acids in all four isolates was proved to be temperature-dependent. The strain Kocuria sp. 301 showed high resistance to all studied extreme conditions (UV radiation, desiccation, and free radicals in medium). The results suggest that isolates from radioactive springs might have developed mechanisms that help them survive under several extreme conditions and could be used in biotechnological production.

Multifarious Indigenous Diazotrophic Rhizobacteria of Rice (Oryza sativa L.) Rhizosphere and Their Effect on Plant Growth Promotion

A diverse group of rhizobacteria persists in the rhizospheric soil, on the surface of roots, or in association with rice plants. These bacteria colonize plant root systems, enhance plant growth and crop yield. Indigenous rhizobacteria are known to promote soil health, grain production quality and serve as sustainable bioinoculant. The present study was aimed to isolate, identify and characterize indigenous plant growth promoting (PGP) diazotrophic bacteria associated with the rhizosphere of rice fields from different areas of Jammu and Kashmir, India. A total of 15 bacteria were isolated and evaluated for various PGP traits, antagonistic activity against phytopathogens, production of hydrolytic enzymes and biofilm formation under in-vitro conditions. The majority of the isolated bacteria were Gram-negative. Out of 15 bacterial isolates, nine isolates produced IAA (12.24 ± 2.86 to 250.3 ± 1.15 μg/ml), 6 isolates exhibited phosphate solubilization activity (36.69 ± 1.63 to 312.4 ± 1.15 μg/ml), 7 isolates exhibited rock phosphate solubilization while 5 isolates solubilized zinc (10–18 mm), 7 isolates showed siderophore production, 8 isolates exhibited HCN production, 6 isolates exhibited aminocyclopropane-1-carboxylate (ACC) deaminase activity, 13 isolates exhibited cellulase activity, nine isolates exhibited amylase and lipase activity and six isolates exhibited chitinase activity. In addition, 5 isolates showed amplification with the nifH gene and showed a significant amount of nitrogenase activity in a range of 0.127–4.39 μmol C2H4/mg protein/h. Five isolates viz., IHK-1, IHK-3, IHK-13, IHK-15 and IHK-25 exhibited most PGP attributes and successfully limited the mycelial growth of Rhizoctonia solani and Fusarium oxysporum in-vitro. All the five bacterial isolates were identified based on morphological, biochemical and 16S rDNA gene sequencing study, as Stenotrophomonas maltophilia, Enterobacter sp., Bacillus sp., Ochrobactrum haematophilum and Pseudomonas aeruginosa. Rice plants developed from seeds inoculated with these PGP strains individually had considerably higher germination percentage, seed vigor index and total dry biomass when compared to control. These findings strongly imply that the PGP diazotrophic bacteria identified in this work could be employed as plant growth stimulators in rice.

Multiplex Lateral Flow Immunoassay for the Detection of Expanded-Spectrum Hydrolysis and CTX-M Enzymes

Background: Early detection of expanded-spectrum cephalosporinase (ESC) hydrolyzing ß-lactamases is essential for antibiotic stewardship. Here we have developed a multiplex lateral flow immunoassay (LFIA) that detects cefotaxime-hydrolyzing activity as well as the most prevalent ESC-hydrolyzing ß-lactamases: the CTX-M-like. Methods: The Rapid LFIA ESC test was evaluated retrospectively on 188 (139 Enterobacterales, 30 Pseudomonas spp. and 14 Acinetobacter spp.) agar-grown bacterial isolates with well-characterized ß-lactamase content. One single colony was resuspended in 150 µL extraction buffer containing cefotaxime, incubated at room temperature for 30 min prior to loading on the LFIA for reading within 10 min. Results: Out of the 188 isolates, all 17 that did not express a β-lactamase hydrolyzing cefotaxime gave negative results, and all 171 isolates expressing a β-lactamase known to hydrolyze cefotaxime, gave a positive test result. In addition, all 86 isolates expressing a CTX-M-variant belonging to one of the five CTX-M-subgroups were correctly identified. The sensitivity and specificity was 100% for both tests. Conclusions: The results showed that the multiplex LFIA was efficient, fast, low cost and easy to implement in routine laboratory work for the confirmation of ESC hydrolyzing activity and the presence of CTX-M enzymes.

Culturable Bacterial Isolates from Arctic Soil shows High Biotechnological Potential

Polar microbiology remains as the most fascinating area of research which mainly focuses on exploration of psychrophilic organisms for having their cold-active enzymes of biotechnological potential. In this study, we have explored a culturable bacterial community and isolated 27 bacterial isolates with a different morphology from an unexplored site of Arctic region, for the possibility of identifying various active biomolecules. Screening of various isolates in a culture dependent manner helped us to identify strains capable of producing extracellular enzymes. The optimal growth parameters of most of the isolates are ranges between 18-22°C temperature, 3-5 days of incubation, 6-9 pH, and 3-5% (w/v) NaCl in LB media. It has also been found that among these isolates, 63% are able to produce lipase, 17% amylase, 7% xylanase and 7% isolates have responded for phosphatase activity but there are no isolates found for gelatinase and cellulase production ability. In addition, few isolates can also produce secretory protease, urease, β-galactosidase, etc. 16SrRNA gene sequence-based phylogeny revealed that the isolates belong to the genera of Psychrobacter, Planococcus, Halomonas, Arthrobacter, Oceanisphaera, Marinbacter, Pseudomonas, Algoriphagus. Strikingly, none of the Arctic isolates showed resistance towards commonly used antibiotics which indicates that the unexplored habitat is devoid of antibiotic exposure and so does the rise of antimicrobial resistance. The structure-function relationship of the isolated bioactive compounds from these isolates are the major focus of future research.

Characterization and Rate of Killing of Conjugated Silver Nanoparticles Against Selected Clinical Bacterial Isolates

The menace of drug resistance, bioavailability and drug delivery to the target sites has motivated researchers to search for new antimicrobial agents from medicinal plants and subsequently use them for the biosynthesis of silver nanoparticles for effective killing of bacteria challenging to kill using crude extracts. The biosynthesis of silver nanoparticles was done using aqueous extract (AQE) of E<i>uphorbia heterophylla</i>, while characterization and the killing rate of conjugated silver nanoparticles (CA_gNP_s) were carried out using standard methods. The maximum wavelength obtained for CA_gNP_s was 410.33 nm, while the size distribution was 237.8 d.nm. The Fourier Transform Infra-Red result showed O-H (3308.94 cm^-1), which is responsible for stabilising and reducing silver ions, while the Transmission Electron Microscopy revealed the presence of monodispersed spherical shapes CA_gNP_s. The Energy Dispersive Spectroscopy confirmed the presence of silver. There were reductions in the clinical bacterial isolates exposed to CA_gNP_s as the exposure time increased. <i>Escherichia coli</i> was killed between 6-7 h while<i> Salmonella typhimurium</i> was killed at the seven has the value of 0.00 log₁₀ CFU/ml was recorded respectively. However, there were increments in the populations of clinical bacterial isolates in control as the time of exposure increased. Therefore, the study suggests that the CA_gNP_s exhibit intense antimicrobial activity and the potential to be developed as an alternative agent to treat bacterial infections, curb multidrug-resistant bacterial infection, and promote speedy drug delivery to the target sites.