Plant-mediated synthesis of ZnO Nanoparticles (NPs) have multiple advantages over conventional synthetic methods like easy, inexpensive, eco-friendly, nontoxic by-products and no critical conditions of temperature and pressure required. In this study, 9.1 g ZnO NPs were synthesized from 230 mL of 0.2 M Zinc acetate dihydrate and 100 mL of green tea leaves extract at room temperature (25°C). The leaves extract was prepared by heating 10gm dried leaves in 100 mL of deionized distilled water at 80°C for 2 h. The synthesized ZnO NPs were dried at 40°C for 24 h and calcined at 100°C for 1 h. The agar well diffusion method was used to evaluate ZnO NPs for antimicrobial activity of selected pathogenic strains. A clear zo“ne of inhibition was measured; 40.05 mm ± 0.137 for Staphylococcus aureus, 36.15 mm ± 0.304 for Escherichia coli and 40.10 mm ± 0.050 for Aspergillus niger that were comparably better results than standard antibiotic Gentamycin that showed 25 and 26 mm zone of inhibition for Staphylococcus aureus and Escherichia coli respectively. The minimum inhibitory concentration scored as 9.765µg ± 0.00, 9.531µg ± 0.00 and 5000µg ± 0.00 for Staphylococcus aureus, Escherichia Coli and Aspergillus niger respectively, was documented low concentration than reported so far concentrations of green tea ZnO NPs.

Aflatoxin contamination in human food and animal feed is a threat to public safety. Aflatoxin B1 (AFB1) can be especially damaging to poultry production and consequently economic development of Pakistan. The present study assessed the in vitro binding of AFB1 by indigenously characterized probiotic lactobacilli. Six isolates (Lactobacillus gallinarum PDP 10, Lactobacillus reuetri FYP 38, Lactobacillus fermentum PDP 24, Lactobacillus gallinarum PL 53, Lactobacillus paracasei PL 120, and Lactobacillus gallinarum PL 149) were tested for activity against toxigenic Aspergillus flavus W-7.1 (AFB1 producer) by well diffusion assay. Only three isolates (PL 53, PL 120, and PL 149) had activity against A. flavus W-7.1. The ameliorative effect of these probiotic isolates on AFB1 production was determined by co-culturing fungus with lactobacilli for 12 days, followed by aflatoxin quantification by high-performance liquid chromatography. In vitro AFB1 binding capacities of lactobacilli were determined by their incubation with a standard amount of AFB1 in phosphate buffer saline at 37 °C for 2 h. AFB1 binding capacities of isolates ranged from 28–65%. Four isolates (PDP 10, PDP 24, PL 120, and PL 149) also ceased aflatoxin production completely, whereas PL 53 showed 55% reduction in AFB1 production as compared to control. The present study demonstrated Lactobacillus gallinarum PL 149 to be an effective candidate AFB1 binding agent against Aspergillus flavus. These findings further support the binding ability of lactic acid bacteria for dietary contaminants.

This study focuses on isolation of pathogenic bacteria from external and internal surfaces of cockroaches collected from houses and hospitals in Lahore. In total, 240 adult cockroaches were collected from houses and hospitals by hand or using sticky traps and food-bait traps. Cockroach species were identified, and microbial screening was done for external surfaces and gut tracts of cockroaches. Jaccard’s index of similarity, Bray–Curtis’ index of dissimilarity, and Shannon–Wiener’s diversity index were used to measure the bacterial community diversity (all species of bacteria) in each habitat. Relative abundance and frequency were measured for each bacterial species on external and internal surfaces of cockroaches. Among human habitations, two major species of cockroaches were isolated, i.e., Periplaneta americana ( P . americana ) and Blattella germanica ( B . germanica ). Out of 240 cockroaches collected, 167 were P . americana and 73 were B . germanica . In total, 11 bacterial species were isolated, but no different bacterial load in each habitat was observed. The most common diagnostic bacterium isolated from the external surface of cockroaches was found to be Escherichia coli (10.31%). In contrast, the most common isolate from the internal gut tract of cockroaches was found to be Pseudomonas aeruginosa , with relative frequency of 19.96%. Jaccard’s index of similarity of bacterial species found on cockroaches was highest (0.3125) in houses, whereas Bray–Curtis’ index of dissimilarity was highest for hospitals (0.2174). The highest Shannon–Wiener’s diversity index value was found in bacteria on cockroaches collected from the Punjab Institute of Cardiology (2.610632). No Salmonella typhi , Staphylococcus epidermidis , Enterobacter aerogenes , and Streptococcus pneumonia were found in the digestive tract of any cockroach.

Anthrax, a severe zoonotic disease, is infrequently reported in anthrax-endemic regions of Pakistan. Despite clinical reports indicating its presence, particularly cutaneous anthrax, there is insufficient laboratory evidence regarding disease occurrence and environmental persistence. The present study aimed to confirm Bacillus anthracis presence, accountable for animal mortality and human infection, while exploring environmental transmission factors. Between March 2019 and July 2021, a total of 19 outbreaks were documented. Of these, 11 affected sheep/goats in Zhob district and 8 affected cattle/sheep in Bajour Agency. Clinical signs suggestive of Bacillus anthracis outbreak were observed in 11 animals. Blood and swab samples were collected for confirmation. The study followed a One Health approach, analyzing animal, environmental (soil/plant), and human samples. Of the 19 outbreaks, 11 were confirmed positive for anthrax based on growth characteristics, colony morphology, and PCR. Soil and plant root samples from the outbreak areas were collected and analyzed microscopically and molecularly. Cutaneous anthrax was observed in six humans, and swab samples were taken from the lesions. Human serum samples (n = 156) were tested for IgG antibodies against PA toxin and quantitative analysis of anthrax toxin receptor 1 (ANTXR1). Bacillus anthracis was detected in 65 out of 570 (11.40%) soil samples and 19 out of 190 (10%) plant root samples from the outbreak areas. Four out of six human samples from cutaneous anthrax lesions tested positive for Bacillus anthracis. Human anthrax seroprevalence was found to be 11% and 9% in two districts, with the highest rates among butchers and meat consumers. The highest ANTXR1 levels were observed in butchers, followed by meat consumers, farm employees, meat vendors, veterinarians, and farm owners. These findings highlight the persistence of anthrax in the region and emphasize the potential public health risks.