Staphylococcus aureus is an important foodborne pathogen associated to food intoxication and other multiple infections in human being. Its presence in salted food is a serious issue due to its salt tolerance potential. A study was conducted to analyze the presence of enterotoxins producing drug resistance S. aureus in salted sea fish from Gwadar. Freshly persevered samples (n=50) of salted fish were subjected to analyze the presence of S. aureus using 16S rRNA and Nuc genes primers. The isolates were then evaluated for drug resistance and enterotoxins producing potential using specific primers for MecA (methicillin resistance gene), (SEA) staphylococcal enterotoxin A and (SEB) staphylococcal enterotoxin B genes. Total 13/50 (26%) of the samples were found positive for the presence of S. aureus, preliminary confirmed with biochemical profiling and finally with the help of target genes presence. The isolates were found showing 100% resistant to methicillin, which were molecularly confirmed by the presence of MecA gene present in genome. The isolates 5/13 (38%) were positive for SEA and 3/13 (23%) for SEB genes, whereas 2/13 (15%) were confirmed having both SEA and SEB genes in its genome. It was also confirmed that all the isolates were capable to form biofilm over the glass surfaces. It was concluded that the study confirmed the presence of enterotoxigenic methicillin resistance Staphylococcus aurous (MRSA) in salted fish product, that poses gross food safety concern. Preventive and control measures are necessary to handle this serious food safety concern.

Propolis oil (PO) was examined for chemical composition, phenolic and flavonoid content, and antioxidant and antimicrobial potential. Phenolic and flavonoid contents were 2.388±1.116mg GAE/g and 0.579±0.140mg QE/g. Oil showed 64.59±14.59 inhibition of DPPH radical and significant antibacterial activities against target bacteria. Salmonella typhi was found to be highly sensitive (27.23±4.35mm) to PO, compared to Escherichia coli (23.40±3.21), Staphylococcus aureus (21.43±2.80), and Klebsiella pneumoniae (21.26±3.25). The MIC and MBS values of PO were 0.35 and 0.7mg/mL for S. typhi and E. coli, whereas they were 0.7 and 1.4mg/mL for S. aureus. Moreover, the PO was found to be bacteriostatic for K. pneumoniae. Aspergillus flavus was found to be highly sensitive to PO, with an effective growth inhibition percentage of 73, followed by Aspergillus niger (70), whereas Aspergillus parasiticus was less sensitive with 25 growth inhibition. Functional groups in PO were determined with an FTIR spectrophotometer, and alcohol, alkane, aldehydes, alkenes, and ketones groups were found to be present, whereas GC-MS analysis revealed the presence of 27 different medicinal compounds, among which α-copanene (29.85), benzyl benzoate (26.8), 2,4-bis[1-(4-hydroxyphenyl)isopropyl]phenol, acetophenone (14.92), undecylenic aldehyde (7.46), p-linalool (5.9), and ethyl 3-phenylpropionate (4.47) were found in abundance.

Berberis baluchistanica Ahrendt is a medicinal plant potentially known for the treatment of different diseases. The bioactive, antioxidant, nutritional components, and antimicrobial properties of crude ethanolic root extract of Berberis baluchistanica were evaluated in this study. The extract was analyzed for total phenolic, flavonoid, DPPH (2, 2‐diphenyl‐1‐picryl‐hydrazyl) scavenging ability, FRAP (ferric reducing antioxidant power), nutritional, and antimicrobial potentials. The alkaloids, tannins, cardiac glycosides, anthraquinones, coumarin, saponins, phenolics, flavonoids, steroids, and terpenoids were confirmed. The extract possessed DPPH radical inhibition with the IC₅₀ of 1.125 mg/mL and FRAP % reduction activity with IC₅₀ (0.912 mg/mL). Total phenolic 19.897 ± 4.8141 mg GAE/g and flavonoid 12.9876 ± 0.8388 mg QE/g contents were confirmed in the root. The extracts exhibit good antibacterial activity against a broad spectrum of food borne pathogens Pseudomonas aeruginosa, Salmonella typhi, Klebsiella pneumoniae, Escherichia coli, and Staphylococcus aureus. The highest inhibitory activity was against Escherichia coli 23.30 ± 1.16 mm and lowest against Klebsiella pneumoniae 7 ± 0.01 mm. Furthermore, the presence of various phytochemical constituents (plant secondary metabolites) was also confirmed with gas chromatography and mass spectroscopy analysis. Results disclosed the occurrence of more than 70 compounds possessing various medicinal properties supporting the traditional uses of root of Berberis baluchistanica in various medical complications indigenously.

The rise of antimicrobial resistance (AMR) in bacterial pathogens such as Klebsiella pneumoniae (Kp) is a pressing public health and economic concern. The ‘One-Health’ framework recognizes that effective management of AMR requires surveillance in agricultural as well as clinical settings, particularly in low-resource regions such as Pakistan. Here, we use whole-genome sequencing to characterise 49 isolates of Klebisella spp. (including 43 Kp) and 2 presumptive Providencia rettgeri isolates recovered from dairy farms located near 3 cities in Pakistan—Quetta (n = 29), Faisalabad (n = 19), and Sargodha (n = 3). The 43 Kp isolates corresponded to 38 sequence types (STs), and 35 of these STs were only observed once. This high diversity indicates frequent admixture and limited clonal spread on local scales. Of the 49 Klebsiella spp. isolates, 41 (84%) did not contain any clinically relevant antimicrobial resistance genes (ARGs), and we did not detect any ARGs predicted to encode resistance to carbapenems or colistin. However, four Kp lineages contained multiple ARGs: ST11 (n = 2), ST1391-1LV (n = 1), ST995 (n = 1) and ST985 (n = 1). STs 11, 1391-1LV and 995 shared a core set of five ARGs, including bla_CTX-M-15, harboured on different AMR plasmids. ST985 carried a different set of 16 resistance genes, including bla_CTX-M-55. The two presumptive P. rettgeri isolates also contained multiple ARGs. Finally, the four most common plasmids which did not harbour ARGs in our dataset were non-randomly distributed between regions, suggesting that local expansion of the plasmids occurs independently of the host bacterial lineage. Evidence regarding how dairy farms contribute to the emergence and spread of AMR in Pakistan is valuable for public authorities and organizations responsible for health, agriculture and the environment, as well as for industrial development.

Foot-and-mouth disease virus (FMDV) is among the most contagious and economically significant viral pathogens of livestock worldwide. In Quetta, Balochistan, recurrent FMDV outbreaks continue to affect cattle and buffalo populations, yet genomic data on circulating strains remain scarce. This study investigated the molecular characteristics and phylogenetic relationships of FMDV serotypes and topotypes detected during 2021 surveillance in Quetta, Pakistan. Twenty epithelial tissue samples, previously confirmed as FMDV-positive through sandwich ELISA, were analyzed for viral RNA using real-time reverse transcription polymerase chain reaction (RT-PCR). Positive samples underwent VP1 gene sequencing, followed by phylogenetic analysis. FMDV RNA was detected in 80% (16/20; 95% CI: 56%-94%) of samples. Among infected animals, 87.5% were female, 69% were cattle, and most (63%) were aged 1-4 years. The highest number of cases (25%) originated from the Jan Muhammad Road area. Phylogenetic analysis revealed all isolates belonged to FMDV serotype O, topotype Middle East-South Asia (ME-SA). These findings confirm FMDV serotype O (ME-SA) as the dominant strain causing endemic outbreaks in Quetta. Strengthening active surveillance and molecular monitoring systems is crucial for early detection, effective vaccination planning, and regulation of cross-border livestock movement with Afghanistan, Iran, and other regions of Pakistan.