Whole-genome sequencing of multidrug resistance Salmonella Typhi clinical strains isolated from Balochistan, Pakistan.

Introduction: Salmonella enterica serovar Typhi (S. Typhi) is a major cause of morbidity and mortality in developing countries, contributing significantly to the global disease burden. Methods: In this study, S. Typhi strains were isolated from 100 patients exhibiting symptoms of typhoid fever at a tertiary care hospital in Pakistan. Antimicrobial testing of all isolates was performed to determine the sensitivity and resistance pattern. Three MDR strains, namely QS194, QS430, and QS468, were subjected to whole genome sequencing for genomic characterization. Results and Discussion: MLST analysis showed that QS194, belonged to ST19, which is commonly associated with Salmonella enterica serovar typhimurium. In contrast, QS430 and QS468, belonged to ST1, a sequence type frequently associated with S. Typhi. PlasmidFinder identified the presence of IncFIB(S) and IncFII(S) plasmids in QS194, while IncQ1 was found in QS468. No plasmid was detected in QS430. CARD-based analysis showed that the strains were largely resistant to a variety of antibiotics and disinfecting agents/antiseptics, including fluoroquinolones, cephalosporins, monobactams, cephamycins, penams, phenicols, tetracyclines, rifamycins, aminoglycosides, etc. The S. Typhi strains possessed various virulence factors, such as Vi antigen, Agf/Csg, Bcf, Fim, Pef, etc. The sequencing data indicated that the strains had antibiotic resistance determinants and shared common virulence factors. Pangenome analysis of the selected S. Typhi strains identified 13,237 genes, with 3,611 being core genes, 2,093 shell genes, and 7,533 cloud genes. Genome-based typing and horizontal gene transfer analysis revealed that the strains had different evolutionary origins and may have adapted to distinct environments or host organisms. These findings provide important insights into the genetic characteristics of S. Typhi strains and their potential association with various ecological niches and host organisms.

Assessment of the dibenzothiophene desulfurization potential of indigenously isolated bacterial consortium IQMJ-5: a different approach to safeguard the environment.

Biodesulfurization is emerging as a valuable technology for the desulfurization of dibenzothiophene (DBT) and its alkylated substitutes, which are otherwise regarded as refractory to other physical and chemical desulfurizing techniques. The inability of the currently identified pure cultures and artificial microbial consortia due to lower desulfurization rate and product inhibition issues has compelled the researcher to look for an alternative solution. Thus, in the present study, an indigenously isolated microbial consortium was employed to tackle the desulfurization issue. Herein, we isolated several kinds of DBT desulfurizing natural microbial consortia from hydrocarbon-contaminated soil samples by conventional enrichment technique. The most effective desulfurizing microbial consortium was sequenced through illumine sequencing technique. Finally, the effect of the products of the desulfurizing pathway (such as 2-hydroxybiphenyl (2-HBP) and sulfate (SO4-2) was evaluated on the growth and desulfurization capability of the isolated consortium. The outcomes of Gibb's assay analysis showed that six isolates followed the "4S" pathway and converted DBT to 2-HBP. Among the isolates, I5 showed maximum growth rate (1.078 g/L dry cell weight) and desulfurization activity (about 77% as indicated by HPLC analysis) and was considered for further in-depth experimentation. The analysis of 16S rRNA by high-throughput sequencing approach of the I5 isolate revealed five types of bacterial phyla including Proteobacteria, Bacteroidetes, Firmicutes, Patescibacteria, and Actinobacteria (in order of abundance). The isolate showed significant tolerance to the inhibitory effect of both 2-HBP and SO4-2 and maintained growth in the presence of even about 1.0 mM initial concentration of both products. This clearly suggests that the isolate can be an efficient candidate for future in-depth desulfurization studies of coal and other fossil fuels.

Bio-Functional Potential and Biochemical Properties of Propolis Collected from Different Regions of Balochistan Province of Pakistan.

Propolis is a well-known resinous natural substance collected by honeybees (Apis mellifera L.) from plants exudations. Variations in chemical composition of propolis are due to different sources from which it is collected and change in climate and geographical location. In this study, different propolis samples were collected from different regions of Balochistan and examined for its chemical composition, total phenolics and total flavonoid contents, and antioxidant potential by using DPPH radical scavenging assay and antimicrobial activity. Bioactive components analysis revealed the presence of steroids, carbohydrates, flavonoids, coumarins, cardiac glycosides, quinones, anthraquinones, terpenoids, tannins, and phlobatannins at different levels. The total phenolics contents were ranged from 2.9343 ± 1.247 to 6.0216 ± 2.873 mg GAE g-1, and flavonoid contents were found to be 0.1546 ± 0.087 to 0.6586 ± 0.329 mg QE g-1, respectively. The antioxidant ability of each extract was analyzed by their concentration having 50% inhibition (IC 50). The propolis sample P3 possessed lower IC 5027.07 ± 0.73 mg mL-1 with higher % inhibition of DPPH radical, and P8 showed lower % inhibition by having IC 5084.43 ± 2.07 mg mL-1. The antibacterial activity of all samples was analyzed against a wide group of bacteria including Escherichia coli, Staphylococcus aureus, Pseudomonas aeruginosa, and Klebsiella pneumonia and propolis extract (P4) was highly active against Klebsiella pneumoniae with the maximum diameter of zone of inhibition 20.33 ± 1.52 mm, and propolis extract (P3) showed maximum zone of inhibition against Escherichia coli19.06 ± 1.90, while propolis extract (P2) was found less active with minimum diameter of zone of inhibition 7.46 ± 1.50 mm. The antifungal activity of extract was considered as active against the fungal species. Propolis extract (P3) showed 82% of zone of inhibition against Aspergillus Niger, and propolis extract (P1) was highly active against Aspergillus parasiticus with 80% of zone of inhibition. By comparing the vibration frequencies in wave numbers of the sample spectrograph acquired from an FTIR spectrophotometer, the functional groups present in the extracts were identified. The presence of seven elements (Fe, Zn, Mn, Ni, Pb, Cd, and Cr) was analyzed through atomic absorption spectrophotometer. The obtained concentrations were within the permissible ranges established by the World Health Organization. The GC-MS analysis revealed the presence of 80 different compounds belonged to different classes. The obtained results confirmed the imperative potential of propolis which can be used in various biological applications.

Meloxicam loaded hydroxypropyl methylcellulose (HPMC) microparticulate: Fabrication, characterization and in vivo pharmacokinetic assessment.

Meloxicam (MEL) is an oxicam derivative with low water solubility that is useful in the treatment of colorectal cancer (CRC) as a COX-2 inhibitor. MEL-loaded HPMC micro particles were fabricated using an oil-in-oil (o/o) emulsion solvent evaporation (ESE) method. FTIR, XRD, particle size analysis, DSC, SEM and in vitro dissolution investigation were utilized to evaluate the produced micro particles physiochemically. Finally, rabbits were used as animal models in an in vivo pharmacokinetic study to assess the MEL concentration in the plasma of rabbits. Pure MEL, F1 and F2 were given to rabbits by a single dose for in vivo pharmacokinetic investigations. The XRD and DSC results confirmed the transformation of MEL from its crystalline nature to the amorphous state in micro particles. The formulations F1 and F2 particle sizes were determined 92.43µm and 163.26µm, respectively. The prepared micro particles had a smooth, non-porous and spherical surface. In comparison to the pure drug (22.4%), the F1 and F2 cumulative drug release (%) was 86.19% and 79.57%, respectively. Pure MEL, F1 and F2 have estimated Cmax values of 7.21, 25.41 and 22.38µg/mL, respectively. MEL had a half-life of 19.98 hours, which rose to 22.19 hours and 24.75 hours for F1 and F2, respectively. MEL, F1 and F2 had AUC0-α values of 116.034, 445.95 and 462.72µg/mL*h, respectively. Considering these aspects, MEL-loaded HPMC micro particles may have the potential to better the delivery and control the release of drug that is not easily dissolved in water which could lead to improved therapeutic efficacy and limited side effects.

Participatory appraisal for healthcare and welfare management strategies of donkeys (Equus ascinus) in Balochistan, Pakistan.

In spite of the significant importance of the donkeys (Equus ascinus) as draft animal in resource-poor countries like Pakistan, they are equines not receiving the appropriate care. They face challenges including injuries, diseases, lack of basic environment and mismanagement by their owners. The present study aims to provide a brief update on the current status of management of healthcare and the welfare of domestic donkeys using participatory epidemiological tools. These tools can help to provide better strategies for improving their productivity and inclusion in human society. This study was mainly focused only on donkeys and horses, mules and ponies were excluded from the study. We carried out a systematic review of the relevant available published literature and shortlisted 50 articles reporting on the different health related characteristics of donkeys. A comprehensive questionnaire was completed by 191 donkey owners, including nine farriers (all men, average age = 38.24 ± 12.43) over a time span from October 2021 to March 2022. Multivariate Odds Ratios (MORs) and 95% confidence intervals were used to assess the predictions of health management and welfare measures for the surveyed donkeys. The most common observed medical problems in donkey health were hyperlipaemia (28.06%), lameness (16.33%) and dental (20.41%) problems. One-third (34.31%) of the donkeys were underweight. The dull donkey with poor appetite needs a clinical emergency owing to a high risk of developing hyperlipemia, which may be life-threatening. These findings are quite useful for the improvement of healthcare management and the welfare of donkeys.

Response of mixed bacterial culture towards dibenzothiophene desulfurization under the influence of surfactants and microscopically (SEM and TEM) characterized magnetic Fe3 O4 nanoparticles.

Excessive emission of sulfur dioxides from the combustion of coal and other fossil fuels for thermal and industrial purposes has been associated with serious environmental hazards. Biodesulfisation (BDS) can be an effective approach for reducing the impact of toxic gases to its inbuilt operational feasibility under ambient environmental conditions. In the present research, two strategies for BDS of a standard organosulfur compound such as dibenzothiophene (DBT) were investigated under laboratory conditions. In the first treatment, the role of different surfactants such as Tween-20, Tween-80, SDS, and EDTA on the desulfurization of DBT was investigated by the application of bacterial consortium IQMJ-5. In the second treatment, Iron oxidenanoparticles were synthesized and immobilized on the surface of bacteria cells. Shake flask experiments were conducted with immobilized cells, surfactant amended immobilized cells, and control or noncoated cells. Among different surfactant treatments, Tween-80 was found to be the most effective surfactant, showing maximum desulfurization activity at a concentration of 5 g/L. The transmission electron microscopy and X-ray diffraction analysis indicated that produced nanoparticles were spherical in shape with a size of about 46 nm and had a stoichiometric ratio of 55.85% and 44.15% between O and Fe, respectively. The nanoparticle treatment enhanced the DBT desulfurization process up to 11.37% as compared to the control, specifically when immobilized cells were used. Therefore, it was concluded that nanoparticles treatments with immobilization of the bacterial cells enhanced the desulfurization rate of DBT under ambient reaction conditions and provide a sustainable alternative for commercial coal BDS.

Comparative analysis of antioxidant activity, toxicity, and mineral composition of kernel and pomace of apricot (Prunus armeniaca L.) grown in Balochistan, Pakistan.

The present study aims to investigate some physical attributes, total phenolics content, total flavonoids content, mineral composition, bioluminescence toxicity assay and antioxidant activity in terms of DPPH, HPS, TAC and FRAP assays in the kernel and pomace samples of six apricot cultivars grown in Balochistan, Pakistan. TFC and TPC determined by the AlCl3 and Folin-Ciocalteu assays in apricot kernel extracts of six cultivars varied from 1797.5 (Chagali) to 4778.9 (Badoghur) mg QUE/100 g DW and from 1750.0 (Chagali) to 5005.8 (Badoghur) mg GAE/100 g DW. Apricot kernels exhibited higher antioxidant activity than pomace; antioxidant activity in terms of IC50 in kernels ranged from 24.88 to 98.61 µg/ml for DPPH, 334.84 to 516.63 µg/ml for HPS, from 22.02 to 110.80 µg/ml for TAC and from 96.27 to 163.35 µg/ml for FRAP. The apricot kernels showed higher TPC, TFC, bioluminescence toxicity to V. logei and antioxidant activity than the pomace. The correlation analysis demonstrated substantial contributions of polyphenols and flavonoids to antioxidant assays. The sample type was the leading factor affecting the amounts of K, Na, Ca, Fe, and Mn in the tested samples; mineral contents were higher in pomace than kernels. The highest inhibition to V. logei was found in the kernels of Badoghur (IC50 = 1.61 mg/ml). The PCA analysis showed significant contributions of phenolic and flavonoid contents towards antioxidant bioluminescence toxicity assays. Our results suggest Badoghur, Shakarpara and Sardai kernels are rich sources of secondary metabolites and possess remarkable antioxidant and antiluminescence activity and can make a significant contribution to the treatment and prevention of chronic health problems.

In Vitro Anti-Inflammatory, Anticancer (MCF-7, 3T3, and HeLa Cell Lines), and Brine Shrimp Lethality Assay and FTIR Analysis of the Extract and Fractions of the Whole Plant of Heliotropium europaeum.

In this study, anti-inflammatory, anticancer, brine shrimp lethality, and FTIR studies were evaluated. The oxidative burst assay using the chemiluminescence technique, MTT assay, brine shrimp lethality assay, and FTIR analysis were the methods used for the evaluation of anti-inflammatory, anticancer, brine shrimp lethality, and FTIR studies, respectively. The whole-plant butanol fraction of Heliotropium europaeum (WBFHE) showed anti-inflammatory activity on ROS having IC5014.7 ± 2.5 while the extract and other fractions of the whole plant of Heliotropium europaeum exhibited no anti-inflammatory activity. None of the extract and fractions of the whole plant of Heliotropium europaeum exhibited anticancer (MCF-7, 3T3, and HeLa cell lines) activities. The whole-plant aqueous fraction of Heliotropium europaeum (WAFHE) and whole-plant butanol fraction of Heliotropium europaeum (WBFHE) showed lethality at high concentration while at low concentration, no toxicity was shown. The whole-plant methanolic extract of Heliotropium europaeum (WMEHE) and whole-plant n-hexane fraction of Heliotropium europaeum (WHFHE) exhibited no toxicity. FTIR interpretation showed the functional groups for the aromatic compounds, phenols, carboxylic acids, esters, alkanes, alkenes, alcohols, alkyl halides, sulfate esters, phosphines, silanes, nitriles, thiols, amines, phosphoric acids, and nitro compounds.

In Vitro Anticancer MCF-7, Anti-Inflammatory, and Brine Shrimp Lethality Assay (BSLA) and GC-MS Analysis of Whole Plant Butanol Fraction of Rheum ribes (WBFRR).

In this study, GC-MS analysis has shown that whole plant butanol fraction of rheum ribes (WBFRR) comprises of 21 compounds which exhibited anticancer (MCF-7) activity having IC50 value of 36.01± 0.26. MTT assay (MCF-7), Oxidative Burst assay using chemiluminescence technique, and B-Hatching techniques were the methods used for anticancer MCF-7, anti-inflammatory, and Brine Shrimp Lethality Assay (BSLA). GC-MS was used for structural elucidation. Whole plant methanol extract of rheum ribes (WMERR), whole plant n-hexane fraction of rheum ribes (WHFRR), and whole plant aqueous fraction of rheum ribes (WAFRR) were inactive against anticancer (MCF-7) cell line. Whole plant methanol extract of rheum ribes (WMERR), whole plant aqueous fraction of rheum ribes (WAFRR) and whole plant butanol fraction of rheum ribes (WBFRR) showed anti-inflammatory activity on ROS having IC50 value of 23.2±1.9, 24.2±2.7 and 12.0±0.6. Whole plant butanol fraction of rheum ribes (WBFRR) showed Brine Shrimp Lethality with LD50 693.302 while whole plant methanol extract of rheum ribes (WMERR) and whole plant aqueous fraction of rheum ribes (WAFRR) showed high lethality at highest concentration. This study revealed that whole plant butanol fraction of rheum ribes (WBFRR) exhibited significant anticancer (MCF-7) activity. In the near future, the constituent of whole plant butanol fraction of rheum ribes (WBFRR) can be the alternative drug against MCF-7 cell line with least toxicity and side effects.