The epidermal mucus of fish plays an important role in innate immunity, providing both physical and biochemical protection through a complex matrix of bioactive molecules. The present study explored the biochemical characterization, bactericidal, and fungicidal activities of Mastacembelus armatus (Zig-Zag eel) epidermal mucus treated with Aeromonas hydrophila. Extracts of epidermal mucus (ethanolic and acidic) of Mastacembelus armatus showed antimicrobial activity against selected resistant bacteria and fungi. The zone of inhibition (ZOI), which was measured in millimetres, revealed differences in activity between different fungi and bacteria. The acidic extract demonstrated noticeably higher ZOI values against various MDR bacteria and fungi was more effective than the ethanolic extract. Strong peaks in the Fourier transform infrared (FTIR) spectrum of Mastacembelus armatus indicated the presence of metabolic components in the epidermal mucus. The O-H and N-H stretching vibrations, which are indicative of water content and protein-based molecules such as mucins or amide, are shown by a large peak at 3272.60 cm-1. This result suggests that fish epidermal mucus has the potential to contribute enormously to the defence of fish against bacteria and fungi. Therefore, this study could be evidence in favour of the substitution of chemical antibiotics with fish epidermal mucus as a new 'antimicrobial' introduced in aquaculture and in humans against bacterial infections.

Background:Acinetobacter baumannii is an important opportunistic pathogen responsible for healthcare-associated infections, particularly among critically ill, intensive care unit (ICU), and post-surgical patients. The emergence of carbapenem-resistant A. baumannii (CRAB) has become a major therapeutic challenge worldwide because of its multidrug-resistant nature and limited treatment options. Despite the increasing prevalence of CRAB in Pakistan, information regarding the molecular characterization of carbapenem resistance genes among isolates recovered from surgical site infections (SSIs) remains limited. Methods: A cross-sectional study was conducted from November 2023 to February 2024 at Khalifa Gul Nawaz Hospital, Bannu, Khyber Pakhtunkhwa, Pakistan. A total of (n = 118) surgical wound specimens were collected from patients with clinically diagnosed SSIs. A. baumannii isolates were identified using standard biochemical tests and confirmed by 16S rRNA gene sequencing. In vitro antimicrobial susceptibility was determined by Kirby-Bauer disk diffusion according to CLSI 2024 guidelines. In addition, the concentration-dependent inhibition-zone response of imipenem and meropenem against carbapenem-resistant isolates was evaluated using an agar well diffusion assay. Molecular detection of blaNDM-1 and blaOXA-23 was performed by polymerase chain reaction (PCR), and representative amplicons were subjected to Sanger sequencing for further analysis. Results: In the current study, the total number of (n = 118) wound specimens from healthcare-associated patients were processed, in which A. baumannii-positive isolates (n = 23, 19.5%) were documented. Its prevalence was higher in males (69.6%) compared to females, and was strongly associated (78.2%) with elderly patients (aged 51-68 years). In vitro susceptibility testing revealed that 91.3% of isolates harbored multidrug-resistant (MDR) attributes. Antimicrobial susceptibility profiling revealed high levels of multidrug resistance, with 82.6% of isolates resistant to imipenem and meropenem, as well as 100% resistance to aztreonam and gentamicin. Colistin showed the highest activity, with 87.0% of isolates remaining susceptible. In the agar well diffusion assay, measurable inhibition of the carbapenem-resistant isolates by imipenem and meropenem occurred only at higher concentrations. Molecular screening of resistance genes identified blaOXA-23 in 60.9% and blaNDM-1 in 30.4% of isolates. Co-expression of both genes was detected in some (n = 4) isolates. Among the selected MDR isolates included in the gene resistance analysis, imipenem and meropenem resistance were recorded in 9/14 blaOXA-23-positive isolates and 5/7 blaNDM-1-positive isolates. Conclusions: This study substantiates an escalated incidence of carbapenem-resistant, MDR A. baumannii in post-surgical infections. blaOXA-23 were documented as the predominant carbapenemase gene, with co-expression of blaNDM-1 in a substantial proportion of isolates. These findings provide an important phenotypic and molecular characterization of selected carbapenemase genes in healthcare-associated infections, highlighting the exigency for strengthened antimicrobial stewardship and infection control strategies in the hospitals of the region.

Chicken manure and its potential to contaminate water systems through the dispersal of pathogenic bacteria are major concerns in environmental and public health. In this study, a metagenomic analysis was employed to systematically identify and compare bacterial assemblages in chicken manure (CM) and in a contaminated sample of chicken manure wastewater (CMW). Whole DNA was extracted from CM and CMW, followed by whole-genome shotgun sequencing; data analysis was done using online Galaxy software (ver. 26.0.1.dev1). Metagenomic analysis reveals a complex One Health challenge. Data showed that CM and CMW are different in their microbiota, as indicated by a distinct separation of beta diversity values and limited overlapping of species between sample types. In the current study, we found a greatly significant common functional set of adapted bacterial masses, including major pathogenic bacterial groups as well as opportunistic and environmental bacterial species, indicative of a direct contamination from CM and CMW. Notably, in both CM and CMW, a plethora of opportunistic, enteric, and environmental pathogens like Escherichia coli, Salmonella enterica, and Acinetobacter baumannii were found, coupled with an indication of a direct functional flow between both ecosystems as tangled reservoirs. Chicken manure samples showed differences in taxonomic composition and inferred functional profiles at the time of sampling: CM1 was pathogen-enriched, CM2 exhibited strong nitrogen-supportive metabolism, CM3 was dominated by fiber-degrading decomposers, and CM4 showed high methane-producing potential with environmental risk. Such findings underscore the raising of chickens as a potential source of harmful bacteria for the environment. It is important to note that this study represents a preliminary investigation with certain limitations, including the absence of biological replicates, lack of temporal sampling, and limited capacity to infer dynamic ecological interactions. Yet this metagenomic report is more about describing the taxonomy and functional potential of the bacteria, rather than discussing the actual ecological processes of these microorganisms in the environment. Future studies will be required to explore these aspects.

Poultry manure is used as fertilizer in most parts of the world. It can harbor diverse nature of pathogenic bacteria and may be one of the active sources of resistant pathogens transmissions. Current study was planned to identify the pathogens as well as their antibiotic resistance profile being isolated from chicken's manure. In present study total (n = 60, 100%) fresh fecal samples were collected from commercial chicken's farms of District Kohat. Samples were initially cultured on different agar media and were incubated at 37 degrees C overnight. Further antibiotics susceptibility and characterization of resistance genes were performed for all the selected pathogens. Only (n = 50, 83.3%) of the total isolates showed proper growth. Most frequent pathogens were Escherichia coli (19, 38%) followed by Salmonella (13, 26%), Klebsiella (8, 16%), Shigella (7, 14%) and Pseudomonas (3, 6%). E. coli revealed highest resistance to doxycycline (16, 84.2%), followed by ampicillin (15, 78.9%), amoxicillin (14, 73.7%) and ciprofloxacin (13, 68.5%). Salmonella showed complete resistance to doxycycline (13, 100%) and tetracycline (11, 84.6%), ciprofloxacin (5, 38.5%) and amoxicillin (4, 31.8%). Pseudomonas were completely resistance to ampicillin followed by levofloxacin (2, 66.7%) and ciprofloxacin (1, 33.3%). Klebsiella showed highly resistance to tetracycline (8, 100%), levofloxacin (6, 75%), ciprofloxacin (4, 50%) and amoxicillin (4, 50%). Shigella strains were completely resistant to doxycycline (7, 100%) and ampicillin (6, 85.7%) while showed less resistance to ciprofloxacin (3, 42.8%) and amoxicillin (2, 28.6%). In the present study, four classes of antibiotics resistance genes (tetA, sul1, str and aadA) were identified with different frequency. The highest frequency (100%) was found for tetA gene followed by the sul1 and str genes (66.6%) each. The lowest frequency (16.6%) was noted for (aadA) gene. It has been concluded that all the isolated pathogens were highly resistant to doxycycline, amoxicillin and erythromycin. To restrict the transmission of resistant pathogens in the environment, it is needed to dispose of poultry manure in proper way, to educate staff of personal hygiene and justified use of antibiotics.

Fish skin mucus contains immunological and antibacterial peptides that protect from foreign invaders. Comprehending the immune response of fish and the interactions between hosts and bacteria may aid in the production of novel approaches to expand disease prevention and control. Therefore, the current study reported proteomic profiling of the skin mucus of Cyprinus carpio, before and after infection with Escherichia coli. In addition, this study emphasized the peptides and proteins in the skin mucus of C. carpio that quantitatively modified against E. coli infection. A total of 2882 comparable proteins (DEPs) at 48 h post-infection were identified by LC-MS/MS (Tandem mass-based Liquid Chromatography Mass Spectrometry). The quantification based on TMT showed that those 375 proteins were Differentially Expressed Proteins (DEPs), of which 102 proteins were downregulated while 273 were upregulated compared to both the treated vs Control groups. Functionally, the 375 DEPs were mostly immune-related, bactericidal cytoskeleton proteins. Based on their ratio, several proteins were substantially upregulated, including ribosomal protein (the 40S, 60S, S3a, L7a, L14, L36), Plastin 1 (I isoform), Calpain 2 (m/II) large subunit b, U6 snRNA-associated Sm-like protein LSm5, Annexin, the homolog of cell division control protein 42, and Keratin such as type I cytoskeletal proteins (K-13, K-15, K-18, and K-19), Conversely, Dedicator of Cytokinesis was one of the downregulated proteins. Functional annotations of DEPs showed their association with cellular function, the innate immune system, stress, and metabolic processes. Our understanding of the interaction between C. carpio and bacteria is improved by this study, which opens the door for further study on potent proteins of fish skin mucus as cutting-edge weapons against antibiotic resistance in aquaculture.

Glutaminase is an enzyme that catalyzes the hydrolysis of glutamine to ammonia and glutamate. Although it can be isolated from bacteria, fungi, plants, and animals, microorganisms remain the preferred source due to their biochemical diversity and ease of cultivation. In this study, 34 soil samples were collected from agricultural waste areas in district Kohat to isolate glutaminase-producing bacteria. Primary screening using phenol red indicator and subsequent biochemical characterization identified 10 positive isolates (30% positivity rate). Among these, four representative strains (A1, G2, P3, and 1R) were selected for detailed analysis, with the majority belonging to the Bacillus genus. Optimal enzyme activity was observed at neutral pH (7.0) and 37°C. SDS-PAGE (12%) analysis was used to estimate the molecular weight of the purified glutaminase. 16S rRNA gene sequencing identified isolate A1 as Bacillus albus, isolates G2 and P3 as closely related to Bacillus anthracis, and isolate 1R as Alkalibacillus bogoriensis. Phylogenetic analysis revealed that Alkalibacillus bogoriensis (1R) clustered distinctly from other Bacillus species. Among all isolates, Bacillus albus demonstrated the highest glutaminase activity, showing good pH and temperature stability. These findings suggest that the isolated strains, particularly Bacillus albus, have potential for future pilot-scale studies and industrial applications in glutaminase production.

BACKGROUND:Helicobacter pylori (H. pylori)is a Gram-negative, microaerophilic, and spiral shape bacterium that resides inside the human stomach. The human stomach serves as its primary reservoir. Complaints about stomach complication due to H. pylori infections are reported in the majority of populations around the globe. Chronic gastritis and intestinal metaplasia of the gastric mucosa are major complications of a long-term H. pylori infections that can lead to gastric cancer in severe cases. This study aims to characterize H. pylori isolates from gastroenteritis patients and to determine the resistance of H. pylori to various antibiotics. METHODS:In the current study, a total of (n = 80) gastric biopsy samples were randomly collected from gastroenteritis patients in brain-heart infusion broth. These were inoculated on Columbia blood agar supplemented with Helicobacter pylori selective supplement (DENT). After culturing, Microscopy and biochemical tests were performed. The susceptibility profile of H. pylori isolates was evaluated using the Kirby Bauer disk diffusion method. On the basis of the drug resistance profile, a total of (n = 20) isolates including (n = 10) from females and (n = 10) from males were selected for the detection and characterization of resistant genes. After confirmation of H. pylori using 16s rRNA, polymerase chain reaction (PCR) was done for the detection of resistance genes including Metronidazole resistance (rdxA gene), Clarithromycin resistance (23s rRNA gene) and Amoxicillin resistance (Penicillin-binding protein A1 (pbpA1) gene). RESULTS:In a total of (n = 80) samples, H. pylori was isolated from 72.5% (n = 58) samples. Among the positive patients, there were 62% (n = 36) of female positive patients while in males, its ratio was 38% (n = 22). It was more common in the age between 30-50 years 55.17% (n = 32). It has shown the highest resistance towards Metronidazole 90% (n = 52), and the lowest toward Levofloxacin 65% (n = 38). Metronidazole resistance gene (rdxA gene) was detected in (n = 13) isolates including (n = 9) isolates from females and (n = 4) from males. In the case of, the Clarithromycin resistance gene (23s rRNA) (n = 10) was positive for H. pylori including (n = 6) isolates from females and (n = 7) were positive for Amoxicillin (pbpA1 gene) including (n = 2) in female and (n = 5) from male patients. CONCLUSION:This study highlights the increasing incidence of H. pylori infections in both male and female patients. It also revealed the current status of antibiotic resistance and its resistance genes in patients facing gastrointestinal issues. Continuous surveillance of resistant clones will help in formulating strategies that can help in combating of resistant clone. It will also help clinician in proper prescription and management of H. pylori infections.

Antibacterial resistance among uro-pathogens has become a global issue for bacterial infection treatment. The antibacterial activity of various plant extracts and silver nanoparticles (AgNPs) derived from medicinal plant extract was evaluated against different drug-resistant uro-pathogens isolated from clinical patients by Agar Well Diffusion and Minimum Inhibitory Concentration (MIC) assays. A total of 100 urine samples were collected from patients admitted to the hospital followed by the identification of Pseudomonas aeruginosa, Escherichia coli, Staphylococcus aureus, Proteus mirabilis, Enterobacter aerogenes, and Klebsiella pneumonia through colony morphology, gram staining, and biochemical characterization. All isolates were highly resistant to all the tested drugs except Amikacin, Gentamicin, and Imipenem. Organic extracts of clove have potent antibacterial activity against all drug-resistant isolates. Interestingly P. aeruginosa was sensitive to all inorganic extracts. These results revealed that the organic extracts of medicinal plants showed more consistent antimicrobial activity compared to their aqueous fraction. Furthermore, the green synthesis of AgNPs derived from clove extract showed strong activity against all tested pathogens followed by green synthesis of AgNPs derived from garlic, ginger, kalwonji, turmeric, and thymus extract respectively compared to medicinal plant extract. Moreover, MIC results showed that AgNPs derived from medicinal plant extract indicate higher efficiency in inhibiting bacterial growth than the medicinal plant extract. The current finding shows that AgNPs derived from plant extract can be recommended as a good alternative for treating the resistant uro-pathogens strains as compared to synthetic drugs and will be considered an attractive candidate for future biological applications.

Syphilis, caused by Treponema pallidum, is resurging globally. Molecular typing allows for the investigation of its epidemiology. In Pakistan and other nations, T. pallidum subsp. pallidum has developed widespread macrolide resistance in the past decade. A study at the Peshawar Regional Blood Centre from June 2020-June 2021 analyzed serum samples from 32,812 blood donors in Khyber Pakhtunkhwa, Pakistan, to assess circulating T. pallidum strains and antibiotic resistance. Blood samples were initially screened for T. pallidum antibodies using a chemiluminescent microparticle immunoassay (CMIA). CMIA-reactive samples underwent polymerase chain reaction (PCR) targeted the polA, tpp47, bmp, and tp0319 genes. PCR-positive samples were further analyzed for molecular subtyping using a CDC-developed procedure and tp0548 gene examination. All PCR-positive samples were analyzed for the presence of point mutations A2058G and A2059G in 23S rRNA, as well as the G1058C mutation in 16S rRNA. These mutations are known to impart antimicrobial resistance to macrolides and doxycycline, respectively. Out of 32,812 serum samples, 272 (0.83%) were CMIA-reactive, with 46 being PCR-positive. Nine T. pallidum subtypes were identified, predominantly 14d/f. The A2058G mutation in 23S rRNA was found in 78% of cases, while G1058C in 16S rRNA and A2059G in 23S rRNA were absent. The research found donor blood useful for assessing T. pallidum molecular subtypes and antibiotic resistance, especially when chancres are not present. The prevalent subtype was 14d/f (51.85%), and the high macrolide resistance of 36 (78%) indicates caution in using macrolides for syphilis treatment in Khyber Pakhtunkhwa, Pakistan.

Gibelion catla is an economically important fish species in Asian fish industry due to its suitability in polyculture system and high market value. Despite a valued market fish in Pakistan and serious threats of bacterial infections this fish is facing, studies concerning health status and disease resistance of G. catla are rare. Therefore, this study used label-free quantification liquid chromatography-mass spectrometry (LFQ-LC-MS/MS) and identified infection-biased expression of the protein in the skin mucus of G. catla after Aeromonas hydrophila infection. In total, 1545 proteins in the skin mucus of nontreated fish and 1400 proteins in the treated fish were identified, of which 1335 proteins overlapped between the two groups, 137 differentially expressed proteins (DEPs) were found, where 71 proteins were downregulated and 66 proteins were upregulated. Functionally, the 137 DEPs were immune-related, bactericidal, and cytoskeleton proteins. The most significant and upregulated protein in the skin mucus of treated fish were ribosomal proteins (40S ribosomal protein S19 (20-fold change (FC)), 60S ribosomal L7a (20-FC), 60S ribosomal L12 isoform X1 (21-FC), and 60S acidic ribosomal P0), beta-actin (24-FC), immunoglobulin mu heavy chain (20-FC), glucose-regulated protein 78 (GRP78; 20-FC), epithelial cell adhesion molecule-like protein (20-FC), putative complement c 3.3 (20-FC), 14 kDa apolipo (19-FC), hemoglobin subunit beta-like protein (19.3-FC), annexin A1-like protein (19-FC), proliferating cell nuclear antigen (18-FC), G-type lysozyme (18-FC), and keratins such as type I cytoskeletal 47 kDa-like isoform X1 (23-FC), and type II cytoskeletal 8 (23-FC). In short, our study highlighted the proteins in the skin mucus of fish involved against bacterial infection, which could be possible biomarkers for further functional-based studies. Particularly, the study elaborates the mucosal immune response of G. catla against A. hydrophila infection, which may help in the strategies of prevention and control of bacterial diseases in this fish.

This study reports the bactericidal activity of mucus extracts and biochemical characterization of skin mucus from five Cyprinids, including Labeo rohita, Ctenopharyngodon Idella, Gibelion catla, Hypophthalmichthys molitrix and Cirrhinus mrigala against ten different bacteria extracted from naturally infected fish. The bactericidal activity was measured based on the zone of inhibition (ZOI) and compared against Fosfomycin. Importantly, acidic mucus extracts from five fish species exhibited higher bactericidal activity than organic and aqueous extracts. The acidic skin mucus extracts of C. idella, L. rohita, and G. catla showed higher ZOI against Staphylococcus aureus, Aeromonas hydrophila, and Pseudomonas aeruginosa. The minimum inhibitory concentration (MIC) of acidic mucus extracts from C. Idella, L. rohita, and G. catla was 16 lg/mL against A. hydrophila, P. aeruginosa, and S. aureus.Further, biochemical characterization of mucus extracts showed that protein concentration was high in the acidic mucus extracts from L. rohita, C. idella, and G. catla compared to H. molitrix and C. mirigala followed by carbohydrate and lipid content. These findings suggest that skin mucus from cyprinids could be a potent source of innovative bactericidal components for fish and human-related treatments. & COPY; 2023 The Author(s). Published by Elsevier B.V. on behalf of King Saud University. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).

We report the proteomic profile of Epidermal Mucus (EM) from Labeo rohita and identified the differentially abundant proteins (DAPs) against Aeromonas hydrophila infection through label-free liquid chromatography-mass spectrometry (LC-MS/MS). Using discovery-based proteomics, a total of 2039 proteins were quantified in nontreated group and 1,328 proteins in the treated group, of which 114 were identified as DAPs in both the groups. Of the 114 DAPs, 68 proteins were upregulated and 46 proteins were downregulated in the treated group compared to nontreated group. Functional annotations of these DAPs shows their association with metabolism, cellular process, molecular process, cytoskeletal, stress, and particularly immune system. Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis and Fisher's exact test between the two groups shows that most of the proteins were immune-related, which were significantly associated with the proteasome, phagosome, and Salmonella infection pathways. Overall, this study shows a basic and primary way for further functional research of the involvement of vitellogenin 2, alpha-2-macroglobulin-like protein, toll-like receptors (TLR-13), calpain, keratin-like proteins, and heat shock proteins against bacterial infection. Nonetheless, this first-ever comprehensive report of a proteomic sketch of EM from L. rohita after A. hydrophila infection provides systematic protein information to broadly understand the biological role of fish EM against bacterial infection.

The skin mucus of fish is an important part of the innate immune system, which is poorly understood at the proteomic level. The study established a complete map of the proteins in the skin mucus of Ctenopharangdon idella (C. idella) and discussed the Differentially Expressed Proteins (DEPs) after Aeromonas hydrophila (A. hydrophila) infection. Using Label Free Liquid Chromatography-Mass Spectrometry (LC-MS/MS) analysis, a total of 126 proteins were identified as differentially expressed, 89 proteins of which were upregulated, and 37 proteins were downregulated. Functional annotations of DEPs showed that the upregulated proteins in the skin mucus of the treated group were mostly associated with complement system and cytoskeleton proteins, whereas downregulated proteins were associated with metabolism. The key upregulated immune proteins were transferrin variant C, lysozyme g, annexin A11, 26S proteasome non-ATPase regulatory subunit 8, hypothetical protein ROHU_000884, 60S ribosomal L7a, calpain-2 catalytic subunit-like protein, calpain-9-like protein, complement component C9, complement C3, cathepsin S, cathepsin Z, 14 kDa apolipo, heat shock protein and intelectin, whereas, leukocyte elastase inhibitor, annexin A11, C-factor-like protein, biotinidase isoform X1 and epidermal growth factor receptor substrate 15-like were the downregulated proteins. Moreover, we for the first-time report proteins such as coactosin, lamin-B2 and kelch 12, which were never reported in fish. Our study directly pointing out the possible immunological biomarkers in the skin mucus of C. idella after A. hydrophila treatment. Each of the protein we report in this study could be used as base to establish their mechanism of action during bacterial infection that may contribute to the strategies against bacterial prevention and control in fishes.

This study explored the bactericidal role of the epidermal mucus (EM) of five freshwater Cyprinid fish species namely Ctenopharyngodon idella, Labeo rohita, Catla catla, Hypophthalmichthys molitrix, and Cirrhinus mrigala after treatment with Aeromonas hydrophila. Extracts of EM (crude and acidic) of each species showed bactericidal activity against various Gram -ve (Pseudomonas aeruginosa, Escherichia coli, Aeromonas hydrophila, Edwardsiella tarda, Salmonella enterica, Klebsiella pneumonia, Serratia marcescens, and Enterobacter cloacae) and Gram +ve (Bacillus wiedmannii and Staphylococcus aureus) bacteria compared with standard antibiotics (Fosfomycin). The zone of inhibition (ZOI) was measured in millimetres against antibiotics (Fosfomycin). Variations in bactericidal activity of EM were observed against bacteria from the same and different fish species. The acidic extract was more effective than the crude extract and showed significantly higher ZOI values against various bacteria and Fosfomycin antibiotics. This result shows that fish EM may perform an important role in fish defence against bacteria. Therefore, this study may hint towards the substitution of synthetic antibiotics with fish EM that may be used as a novel 'bactericidal' in aquaculture as well as in humans against bacterial infections.

The ability of microbes to resist or neutralize the action of drugs that have been used against microbes is considered as antimicrobial resistance (AMR). AMR among different strains of Escherichia coli is considered as a major threat to public health. Drug-resistant in E. coli is found predominantly in the hospital sittings, in the community, and surrounding environment. It has adopted different defensive strategies to minimize the effects of drugs. Extended-spectrum β-lactamase (ESBL), fluoroquinolones, and carbapenemases have been considered as strong resistance strategies being present in most of resistant bacterial strains. Mobile genetic elements (MGEs) have the major contribution in the transfer of resistance genes in between or among bacterial cells. Plasmids are normally present in most of resistant strains, helping in the transfer of genetic material between bacterial cells. Transposons another MGEs, are being considered as one of the major sources of resistance transmission. Collectively, MGEs play an important role in facilitating in exchange, acquisition, and dissemination of resistance genes. Resistance in E. coli has been reported worldwide and there is variation in its resistance pattern. CTX-M ESBLs, carbapenems, colistin-resistant, and ST-131 E. coli resistant clones are considered the most dominant phenotypes. The aforesaid resistant variants are predominantly found in densely populated regions, Sub-Saharan Africa, China, and South Asian countries.

This study aimed to elucidate differentially expressed proteins in drug resistant Salmonella Typhi. Among 100 samples, S. typhi were identified in 43 samples. In drug susceptibility profile, 95.3% (41/43), 80% (35/43) and 70% (30/43) resistances were observed against Nalidixic acid, Ampicillin, and Chloramphenicol respectively. No resistance was observed against Imipenum and Azithromycin while only 11% (5/43) isolates were found resistant to Ceftriaxone. Mass spectrometric differential analysis resulted in 23 up-regulated proteins in drug resistant isolates. Proteins found up-regulated are involved in virulence (vipB, galU, tufA, and lpp1), translation (rpsF, rpsG, rplJ, and rplR), antibiotic resistance (zwf, phoP, and ompX), cell metabolism (metK, ftsZ, pepD, and secB), stress response (ridA, rbfA, and dps), housekeeping (gapA and eno) and hypothetical proteins including ydfZ, t1802, and yajQ. These proteins are of diverse nature and functions but highly interconnected. Further characterization may be helpful for elucidation of new biomarker proteins and therapeutic drug targets.

Different resistance mechanisms are involved in exhibiting resistance to different groups of antibiotics. Researchers are searching for new therapeutic options to encounter the emerging trend of microbial resistance. Bacteria were isolated from the extreme environment of Cholistan Desert and were screened for characterization. Potential metabolites that showed broad-spectrum activity were partially purified using silica gel chromatography and determined their minimum inhibitory concentration. A collection of 50 bacterial isolates from soil samples was screened for metabolite production and among them isolate R19 of Bacillus sp. and isolate A8 of Alcaligenes sp. had high similarity with strong antimicrobial metabolite producers. The growth of A8 was stable at slight acidic pH while R19 was best at neutral pH. Similarly, the best growth of A8 was observed at 37 °C while R19 at 35 °C. Minimum inhibitory concentration of purified compounds of Bacillus sp. were determined at concentration range of (3.12 – 100 %) against multidrug-resistant (MDR) strains of Shigella, Enterobacter aerogenes, Staphylococcus aureus, Pseudomonas aeruginosa, Escherichia coli, and Klebsiella pneumonia and produced 10 – 25 mm zone of inhibition. Metabolites of Alcaligenes sp. were sufficient to inhibit the growth of all selected MDR bacteria at concentrations 12.25 – 100 % and shows 10 – 20 mm zone of inhibition. Bacillus sp. and Alcaligenes sp. can be used as producers of potential antibacterial metabolites. Proper utilization of selected metabolites can be helpful in combating emerging drug resistant pathogenic bacteria. In addition, further proteomic analysis and structural insight should be considered to elaborate their active ingredients and its efficacy.

Aims: Thermophilic proteases are important industrial enzymes because they can be used at high temperatures in various bioprocessing schemes. The bacterial population of the Cholistan desert was explored for thermophilic proteases and their industrial applications. Methodology and results: Three bacterial isolates K1, K5 and K7 were found promising protease producers. These isolates were preliminary identified as Bacillus based on morphological characteristics and biochemical tests (positive for catalase, oxidase and citrate tests, and negative for indole and urease tests). The isolates K1, K5 and K7 were further identified as Priestia endophytica, Lysinibacillus cresolivorans and Bacillus subtilis, respectively by phylogenetic analysis. The isolates grew best at 50 degrees C and P. endophytica (K1), L. cresolivorans (K5) and B. subtilis (K7) produced larger zones of hydrolysis at 37 degrees C, 45 degrees C and 50 degrees C at pH 7, respectively. The optimum temperature where protease activity was maximum was 65 degrees C for P. endophytica and L. cresolivorans and 55 degrees C for B. subtilis, and the optimum pH was 9. Conclusion, significance and impact of study: The proteases produced by these isolates were found active at high temperatures (45 degrees C to 85 degrees C) and high pH (9-12), which make them industrially important thermoalkaliphilic proteases. These proteases successfully de-haired cow's skin and de-stained blood from cotton cloth pieces, which are rarely tested applications of these proteases.

Introduction: Helicobacter pylori is an important medical pathogen present in more than half of the world’s population. Various treatment regimen are in use for the eradication of H. pylori, but due to the emergence of antibiotic resistance, its management is a big issue for clinicians. Methods: In this study all suspected cases that had visited District Headquarters Hospital Kohat were considered for screening of H. pylori infections. Preliminary information about their age, gender, general health conditions, occupation, etc. was taken for consideration. After recording initial signs and symptoms, samples were considered for H. pylori detection using stool antigen test and endoscopy. Fourteen-day proton pump inhibitor base triple and quadruple therapy were administered to each patient. Results: In total (n = 178), there were high numbers of positivity in patients aged below 30 years (82; 46.06%), most of whom belonged to rural areas. Conclusion: This study concludes that there were high numbers of positive patients aged below 30 years, and according to this study MEL (Metronidazole + Esomeprazole + Levofloxacin) is the most effective treatment regimen for the eradication of H. pylori.

Background: The genus Proteus is a Gram-negative bacterium with a unique characteristic of swarming. Mainly three species are involved in initiating urinary tract infections in the community and in immunocompromised patients, particularly in patients going through long-term catheterization. Due to their strong virulence factors like biofilm formations, protease, and hemolysin, they can lead to lengthening infections in affected individuals. Probiotics are live bacteria and yeasts that are beneficial to human health and can be used as an alternative for the control of nosocomial diseases. Lactobacilli are one of the common probiotics mostly found in yogurt and other fermented foods that have been used as a substitute for infection control.Objectives: The current study was designed to screen potential probiotic bacteria to encounter antibiotic-resistant and virulent Proteus species.Methods: In the current study, using probiotics, already known antibiotic-resistant isolates (n = 25) of Proteus were processed to characterize their virulence factors and their inhibition. Biofilm formation, protease, and hemolysin activities were studied using different phenotypic detection methods. Further, their virulence genes zapA, flg, hmpA, mrp, and rsbA were explored using their genomic DNA. These isolates were found resistant to different classes of antibiotics, and a strategy was designed to inhibit their growth by using probiotic bacteria isolated from the soil.Results: Virulence factors first, all isolates were subjected to biofilm detection, and they were 32% (n = 8) strong, 40% (n =10) mod-erate, 16% (n = 4) weak, and 12% (n =3) non-biofilm producers. All isolates were positive for swarming activity by showing a differen-tiated ring form of growth. Protease activity showed 56% (n = 14) isolates. Only 24% (n = 6) of isolates were positive for hemolysin. Virulence factors and molecular mechanisms were studied, and gene rsbA responsible for swarming was amplified in 17 (68%) Proteus isolates, and mrp responsible for fimbria was detected in 19 (76%) bacterial isolates. Further, these isolates were subjected to flagella, protease, and hemolysin, and it was revealed that flg 11 (44%), 13 (52%) protease coding zapA, and hmA gene coding hemolysin were amplified in 2 (8%) Proteus isolates. Probiotic bacteria isolated from soil samples were probed for antagonistic activity against Proteus species. The probiotic bacteria were identified as Lactobacillus plantarum, Bacillus subtilis, and B. licheniformis. Due to their strong growth inhibitory effects against Proteus, it is crucial to characterize further the metabolites that have shown suppressive results against Proteus.Conclusions: Findings from the current study will provide new avenues for drug development and also help clinicians manage re-sistant pathogens in healthcare settings. Probiotic applications for infection control can be useful in treating resistant pathogens. Further purification and characterization of metabolites will provide alternative options for managing resistance issues in mi-crobes.