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BACKGROUND: Bovine mastitis results in significant economic losses for the dairy industry globally due to milk production losses and decreased herd efficiency. This research aimed to isolate, select, and characterize indigenous lactobacilli with probiotic properties. A total of 40 lactobacilli were isolated from healthy milk samples of cattle and identified at the species level through 16S rDNA sequencing. All isolates were initially screened for antimicrobial activity, and selected isolates underwent in vitro assessment of probiotic properties. RESULTS: Among the lactobacilli isolates, varying levels of activity (9 to 19 mm) against cattle mastitogens; Stapylococcus aureus (Staph. aureus), Escherichia coli (E. coli) and Streptococcus dysgalactiae (Strep. dysgalactiae) were observed in the well diffusion assay. These isolates demonstrated auto-aggregation (ranging from 14.29 ± 0.96% to 62.11 ± 1.09%) and co-aggregate (ranging from 9.21 ± 0.14% to 55.74 ± 0.74%) with mastitogens after 2 h. Lactobacillus (Lb.) plantarum CM49 showed sensitivity to most antibiotics tested and exhibited strong inhibitory effects, with mean log10 reductions of 3.46 for Staph. aureus, 2.82 for E. coli, and 1.45 for Strep. dysgalactiae in co-culture experiments. Furthermore, Lb. plantarum CM49 significantly decreased the adhesion rate of mastitogens on the bovine mammary cell line and mouse model, demonstrating its potential effectiveness in preventing mastitis. CONCLUSION: It is concluded that Lb. plantarum CM49 has remarkable probiotic potential with activity against cattle mastitogens in the laboratory and cell culture and competitively excludes mastitogens from bovine mammary cells and ameliorates Staph. aureus-induced mastitis in mice.

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Background and Objectives: Acinetobacter baumannii (A. baumannii), particularly carbapenem-resistant A. baumannii (CRAB), represents a grave concern in healthcare settings and is associated with high mortality. This study aimed to conduct molecular, mutational, and phylogenetic analyses of specific genes in CRAB and evaluate the synergistic effects of selected antimicrobial combinations. Materials and Methods: Phenotypic characterization was performed on six CRAB strains by using the Modified Hodge Test (MHT) and IMP-EDTA Double-Disc Synergy Test (IMP-EDTA DDST). Carbapenemase- and metallo-beta-lactamase-encoding genes were amplified by using Polymerase Chain Reaction. Phylogenetic analysis using the MEGA 11 tool was used to determine the evolutionary relatedness of these genes. Mutational analysis was performed by using I-Mutant, MUPro, and PHD-SNP bioinformatics tools to predict mutations in the carbapenemase-encoding genes. Microdilution checkerboard titration assessed the synergistic effects of antimicrobial combinations (azithromycin-meropenem, rifampicin-meropenem, meropenem-colistin, and azithromycin-colistin) on these CRAB isolates. Results: The phenotypic characterization of six CRAB isolates revealed positive results for MHT and IMP-EDTA DDST. The molecular characterization revealed that carbapenemase- and MBL-encoding genes were present in all isolates with varying frequencies, including blaOXA-51 (100%) and blaIMP (0%). The sequence analysis revealed high evolutionary relatedness to sequences in the NCBI database. The mutational analysis identified 16 mutations, of which 1 mutation (P116L) in the blaOXA-58 gene predicted a change in the protein product, potentially contributing to carbapenem resistance. The checkerboard titration method did not reveal any synergism among the tested antimicrobial combinations against CRAB. Conclusion: This study's findings underscore the significant challenges posed by CRAB isolates harboring multiple resistant genes in treatment. This highlights the urgent need for novel antimicrobial agents, a crucial step towards reducing mortality rates not only in Pakistan but also globally.

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This research investigates the impact of sea level rise (SLR) on the Indus Delta, a vital ecosystem increasingly vulnerable to climate change repercussions. The objective of this study is to comprehensively assess the flooded areas under various shared socioeconomic pathway (SSP) scenarios based on the Intergovernmental Panel on Climate Change's (IPCC) 6th Assessment Report. The study employs a GIS-based bathtub model, utilizing historical (1995-2014) and IPCC-projected (2020-2150) tide gauge data from Karachi, Kandla, and Okha stations to identify potential inundated areas threatened by coastal flooding. Additionally, it analyzes LANDSAT-derived multispectral images to identify coastal erosion hotspots and changes in the landscape. A supervised random forest classifier is used to classify major landforms and understand alterations in land cover. Furthermore, neural network-based cellular automata simulations are applied to predict future land cover for 2050, 2100, and 2150 at risk of inundation. The results indicate that under different SSP scenarios, the estimated inundated land area varies from 307.36 km2 (5 % confidence on SSP1-1.9) to 7150.8 km2 (95 % confidence on SSP5-8.5). By 2150, the region will lose over 550 km2 of agricultural land and 535 km2 of mangroves (mean SLR projection). This work emphasizes identifying sensitive land cover for SLR-induced coastal flooding. It might fuel future policy and modeling endeavors to reduce SLR uncertainty and build effective coastal inundation mitigation methods.

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This research paper reports an enhancement of thermal, optical, mechanical and antibacterial activities of the Polyvinyl alcohol-Nanodiamonds (PVA-NDs) composite required for the food packaging industry. The synthesis of composites was done by the wet processing method. The large surface area of NDs facilitated the robust interaction between the hydroxyl group and macromolecular chains of PVA to enhance the hydrogen bonding of PVA with NDs rather than PVA molecules. Thus, a reduction in PVA diffraction peak intensity was reported. NDs improved the thermal stability by preventing the out-diffusion of volatile decomposition products of PVA. The results also revealed an enhancement in tensile strength (∼60 MPa) and ductility (∼180 %). PVA-NDs composite efficiently blocked the UVC (100 %), most of the part of the UVB (∼85 % above 300 nm), and UVA (∼58 %). Furthermore, enhanced antibacterial activities were reported for PVA-NDs composite against E. coli and S. aureus. NDs accumulated around the bacterial cells prevented essential cellular functions and led to death. Hence, this composite could be a promising candidate for safe, thermally stable, strong, flexible, transparent, UV- resistant antibacterial food packaging material.

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Coxiella burnetii is the worldwide zoonotic infectious agent for Q fever in humans and animals. Farm animals are the main reservoirs of C. burnetii infection, which is mainly transmitted via tick bites. In humans, oral, percutaneous, and respiratory routes are the primary sources of infection transmission. The clinical signs vary from flu-like symptoms to endocarditis for humans' acute and chronic Q fever. While it is usually asymptomatic in livestock, abortion, stillbirth, infertility, mastitis, and endometritis are its clinical consequences. Infected farm animals shed C. burnetii in birth products, milk, feces, vaginal mucus, and urine. Milk is an important source of infection among foods of animal origin. This study aimed to determine the prevalence and molecular characterization of C. burnetii in milk samples of dairy animals from two districts in Punjab, Pakistan, as it has not been reported there so far. Using a convenience sampling approach, the current study included 304 individual milk samples from different herds of cattle, buffalo, goats, and sheep present on 39 farms in 11 villages in the districts of Kasur and Lahore. PCR targeting the IS1111 gene sequence was used for its detection. Coxiella burnetii DNA was present in 19 of the 304 (6.3%) samples. The distribution was 7.2% and 5.2% in districts Kasur and Lahore, respectively. The results showed the distribution in ruminants as 3.4% in buffalo, 5.6% in cattle, 6.7% in goats, and 10.6% in sheep. From the univariable analysis, the clinical signs of infection i.e. mastitis and abortion were analyzed for the prevalence of Coxiella burnetii. The obtained sequences were identical to the previously reported sequence of a local strain in district Lahore, Sahiwal and Attock. These findings demonstrated that the prevalence of C. burnetii in raw milk samples deserves more attention from the health care system and veterinary organizations in Kasur and Lahore of Punjab, Pakistan. Future studies should include different districts and human populations, especially professionals working with animals, to estimate the prevalence of C. burnetii.

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Objective: This research aimed to analyze the prevalence, molecular characteristics, toxinotyping, alpha toxin production potential, and antibiotic resistance pattern of Clostridium perfringens (C. perfringens) isolates in meat samples collected from various sources. Methods: Sixty meat samples were screened for alpha toxin using Enzyme-Linked Immunosorbent Assay (ELISA), revealing a positivity rate of 13.3%, predominantly in raw poultry meat. Subsequent culturing on Perfringens agar identified nine samples harboring characteristic C. perfringens colonies, primarily isolated from raw poultry meat. Molecular confirmation through 16S rRNA gene amplification and sequencing authenticated twelve isolates as C. perfringens, with nine strains exhibiting genetic resemblance to locally isolated strains. Toxinotyping assays targeting alpha toxin-specific genes confirmed all nine isolates as type A C. perfringens, with no detection of beta or epsilon toxin genes. Hemolytic assays demonstrated varying alpha toxin production potentials among isolates, with accession number OQ721004.1 displaying the highest production capacity. Moreover, antibiotic resistance profiling revealed multi-drug resistance patterns among the isolates. Results: The study identified distinct clusters within C. perfringens strains, indicating variations. Phylogenetic analysis delineated genetic relatedness among strains, elucidating potential evolutionary paths and divergences. Conclusion: The findings underscore the need for robust surveillance and control measures to mitigate the risk of C. perfringens contamination in meat products, particularly in raw poultry meat. Enhanced monitoring and prudent antimicrobial stewardship practices are warranted in both veterinary and clinical settings to address the observed antibiotic resistance profiles and prevent foodborne outbreaks.

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Tricuspid valve regurgitation, or TR, is a difficult-to-manage condition. In addition to EVOQUE, percutaneous annuloplasty, and surgical repair, the TriClip G4 system has been added to the interventional therapeutic choices for TR. Recently, the Food and Drug Administration (FDA) approved the use of the TriClip G4 device to treat patients with symptomatic, severe TR who have received optimal medication therapy but are at intermediate or higher risk of surgery. This review attempts to offer a thorough examination of the procedural features, learning curves, results of the device and compares the TriClip G4 system to other interventional therapies for TR. TriClip G4 has shown to have promising results in pivotal clinical trials, be cost-effective, and improve the quality of life of patients. Furthermore, it has its unique advantages against other conventional techniques and devices.

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Bacterial antimicrobial resistance (BAMR) seems to pose the greatest threat to public health, food safety, and agriculture in this century. The development of novel efficient antimicrobial agents to combat bacterial infections has become a global issue. Silver nanoparticles (Ag NPs) appeared as a feasible alternative to antibiotics. However, Ag NPs face cost, toxicity, and aggregation issues which limit their antibacterial activity. This work aims to stabilize Ag NPs with enhanced antimicrobial activity at comparatively lower Ag concentrations to prevent bacterial infections. For this purpose, the Ag core was covered with nanodiamonds (NDs). Ag-NDs composite have been synthesized by microplasma technique. TEM analysis confirmed the presence of both Ag and NDs in the Ag-NDs composite. A particle size (∼19 nm) was reported for Ag-NDs at the highest concentration as compared to Ag NPs (∼3 nm). The conduction band of the diamond acted as an extremely strong reducing agent for Ag NPs. The large surface area of NDs stabilized the Ag NPs. A redshift (∼400 nm-406 nm) in UV-visible spectra of the Ag-NDs composite indicated the formation of bigger-sized Ag NPs after incorporating NDs. XRD and LIBS analysis verified the increase in intensity of Ag-NPs by increasing ND concentration. The presence of functional groups including OH, CH, and Ag/Ag2O was confirmed by FTIR. Bacterial inhibition growth appeared to be a dose-dependent process. The minimum inhibition concentration value of Ag-NDs composite at the highest NDs concentration against E. coli (∼ 0.69 µg/ml) and S. aureus (∼44 µg/ml). This is the first study to report the smallest MIC for E. coli (<1 µg/ml). Ag-ND composites emerged to be more efficient than Ag NPs and preferred to be used against BAMR. The enhanced antibacterial activity of the Ag-NDs composite makes it a potential candidate for antibiotics, food products, and pesticides.

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Aims@#The study was aimed to explore the antimicrobial potential of ethanolic leaf extracts of Eucalyptus globulus, Moringa oliefera, Syzygium cumini and Citrus limon against antibiotic-resistant Clostridium perfringens type D (n=5).@*Methodology and results@#Antibiotic resistance pattern of C. perfringens type D isolates against tetracycline, gentamicin, ceftriaxone, amoxicillin and streptomycin was evaluated by disc diffusion method. Well diffusion and micro broth dilution methods were used to determine the anti-bacterial activity, sub-inhibitory concentrations and antibiotic resistance modulating effects of the plant extracts. Ethanolic extract of E. globules was selected to evaluate its modulatory impact and subjected to GC-MS analysis to separate and identify the phytochemicals. The results showed that the isolates were resistant to gentamicin (0 ± 0.00 mm), streptomycin (0 ± 0.00 mm), tetracycline (13.2 ± 2.28 mm) and ceftriaxone (0 ± 0.00 mm) while sensitive to amoxicillin (23.8 ± 1.30 mm) and tetracycline (13.2 ± 2.28 mm). Eucalyptus globulus exhibited the maximum anti-bacterial activity with a zone of inhibition (ZOI) of 14.6 ± 0.54 mm and minimum inhibitory concentration (MIC) (1500 ± 947.85 µg/mL). Other plant extracts (M. oliefera, S. cumini and C. limon) also showed anti-bacterial activity but couldn’t modulate the resistance. The activity of ceftriaxone associated with E. globulus extract was improved with 20.2 ± 0.20 mm ZOI at 78.125 µg/mL sub-inhibitory concentration.@*Conclusion, significance and impact of study: @#The study results indicate the possible use of the ethanolic extract of E. globulus alone or in combination with common antibiotics for the treatment of C. perfringens infections in small ruminants.

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In this study, we checked the effectiveness of L. fermentum IKP 111 in treating S. enteritidis infection in an in vivo study. Its oral administration to broiler chicks significantly reduced the colonization of S. enteritidis in the gut and there was a lower bacterial count of S. enteritidis in the droppings after infection. The administration of the probiotic L. fermentum IKP 111 also led to increase in weight gain in the broiler chicks as well as their immunomodulation against avian influenza virus (AIV) and Newcastle disease virus (NDV) as compared to the chicks challenged only with S. enteritidis. Our study provides evidence that the probiotic strain L. fermentum IKP 111 could be an alternate for controlling S. enteritidis infection while enhancing the gut health as well as the immune response of broiler chickens against viral infections.

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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.

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Background: We determined the prevalence of antimicrobial resistance (AMR) in polymicrobial pathogens in Pakistan. Methods: A total of 70,518 clinical samples were collected aseptically and confirmation of isolates and antibiogram were performed by the VITEK 2 system. Results: Of 70,518 samples, 441 (0.62%) were polymicrobial samples, with 882 (1.2%) polymicrobial pathogens with 689 (78.1%) Gram-negative rods (GNRs), 166 (18.8%) Gram-positive cocci and 27 (3.1%) Candida albicans. Among GNRs, 28.8% were Escherichia coli and 25.9% were Klebsiella pneumoniae. Majority, 15.1% of Pseudomonas aeruginosa and K. pneumoniae were found in combination. 30.1% of isolates were ESBL producers, 9.7% carbapenem-resistant organisms, 35.5% MRSA and 6.0% VRE. 100% of E. coli were resistant to ampicillin and 98% of K. pneumoniae were resistant to piperacillin. Conclusion: A high prevalence of AMR in polymicrobial pathogens was observed.

Infections caused by one or more types of bacteria, viruses, fungi or parasites ­ known as polymicrobial infections ­ are a threat to health. These infections cause serious illness and are linked to high numbers of deaths, long hospital stays and high costs of treatment. Usually, polymicrobial infections are treated with combinations of antimicrobials. However, microbes becoming less susceptible to antimicrobials (known as antimicrobial resistance) is an increasing problem. To find out how common resistance is in Pakistan, this study tested 70,518 clinical samples. Of these, 441 tested positive for polymicrobial infections. These included Candida albicans, Gram-positive cocci and Gram-negative rods infections. Many of these were resistant to widely used antibiotics such as penicillins, cephalosporins, quinolones and fluoroquinolones. This study concluded that hospitals in Pakistan have a high prevalence of resistance and that better cleanliness practices should be put in place to combat this.

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Coxiella burnetii is the zoonotic pathogen that causes Q fever; it is widespread globally. Livestock animals are its main reservoir, and infected animals shed C. burnetii in their birth products, feces, vaginal mucus, urine, tissues, and food obtained from them, i.e., milk and meat. There were previously very few reports on the prevalence of C. burnetii in raw meat. This study aimed to determine the prevalence of C.burnetii and its molecular characterization in raw ruminant meat from the Kasur and Lahore districts in Punjab, Pakistan, as this has not been reported so far. In this study, 200 meat samples, 50 from each species of cattle, buffalo, goat, and sheep, were collected from the slaughterhouses in each district, Kasur and Lahore in 2021 and 2022. PCR was used for the detection of the IS1111 element of C. burnetii. The data were recorded and univariate analysis was performed to determine the frequency of C. burnetii DNA in raw meat samples obtained from different ruminant species using the SAS 9.4 statistical package. Of the total of 200 raw meat samples, C. burnetii DNA was present in 40 (20%) of them, tested by PCR using the IS1111 sequence. The prevalence of C.burnetii differed among the studied species of ruminants. When species were compared pairwise, the prevalence in cattle was statistically significantly lower than in sheep (P = 0.005). The sequence alignment based on origin implied that the strains are genetically diverse in different districts of Punjab, Pakistan. The findings demonstrated that the prevalence of C. burnetii, especially in raw meat samples, deserves more attention from the health care system and professionals from Punjab, Pakistan, i.e., abattoir workers and veterinarians.

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In the community and among hospitalized patients, urinary tract infections (UTIs) rank as the most common bacterial infections. The researchers processed urine samples obtained from affiliated hospitals of Peshawar Medical College. The samples were examined under a microscope to assess the presence of bacteria, pus cells and red blood cells. Following this, the samples were inoculated on MacConkey and blood agar and subsequent antibiotic susceptibility testing was conducted. The findings revealed that 35.9% of hospitalized patients and 16.9% of outpatients were diagnosed with UTIs. Furthermore, 82.2% of the identified UTIs were found to be multidrug-resistant (MDR), with MDR Escherichia coli accounting for 77% of cases. Trimethoprim sulfamethazine (26.8%), penicillin (0%), cefepime (27.8%), cefotaxime (23.7%), aztreonam (2.1%), meropenem (86.6%), ciprofloxacin (51.5%), amoxicillin/clavulanic acid (37.1%), nitrofurantoin (70.1%), gentamycin (73%), ceftazidime (19.5%), levofloxacin (51.5%) and ceftriaxone (25.77%) were subjected to antibiotic susceptibility testing. It is concerning that among the 13 antibiotics examined, solely nitrofurantoin displayed oral efficacy as an effective treatment choice for UTIs.

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Clostridium perfringens toxins play role in causing pulpy kidney disease poisoning as well enterotoxaemia. To combat antimicrobial resistance: curbing use, regulations and execution to antimicrobial usage in food along with withdrawal period is necessary. Aim of study was to optimize the toxins production by indigenously characterized C. perfringens type D isolates (n=03) under various physicochemical parameters, a lead towards local vaccine production in Pakistan. Indigenous isolates were characterized on the basis of 16S rRNA and MW349974.1, MW341428.1, MW332258.1 accession numbers were received from NCBI GenBank. Isolates were identified as toxinotype D through PCR toxinotyping. Quinolones antibiotic susceptibility testing revealed that isolates observed susceptible to enrofloxacin and resistant to ciprofloxacillin and ofloxacillin. Optimization of toxins production was determined under the influence of physical and chemical parameters. Alpha and epsilon toxin production in reinforced clostridial medium (RCM) broth was observed higher at 37°C after 24h incubation by MW332258.1. Under the influence of 0.2% glucose and 0.3% tween 80 supplementation in RCM, greater production of alpha and epsilon toxin units was observed by MW332258.1. Under optimized physicochemical parameters, maximum toxins units were observed; MW332258.1 isolate is excellent candidate could be used to produce maximum toxin units for vaccine production at industrial scale.

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Reverse osmosis (RO) spiral wound membrane generation reached 93.5% in 2020, resulting in 14,000 tons of used RO membranes being discarded annually into landfills, which is unprecedented. The current study aims to chemically convert the end-of-life RO membrane, followed by its performance evaluation and microbial removal efficiency on three different sources of water, i.e., tap water (TW), integrated constructed wetland permeate (ICW-P), and membrane bio-rector permeate (MBR-P), respectively. This was accomplished by selecting 6 years of spent Filmtech (LC-LE-4040) thin film composite type brackish water reverse osmosis (BWRO) membrane, followed by alkaline and acidic cleaning for 2 h. Finally, the conversion was carried out by 6% sodium hypochlorite (NaOCl) with 300,000 ppm/h exposure by active system (AS) using the clean in place CIP pump at 2 bars for 10 h duration. The membrane demonstrated 67% water recovery and 1% saltwater rejection, which means RO membrane now converted into recycled RO (R-RO) or (UF) by removal of the polyamide (PA) layer. Water recovery was 67% for TW, 68% for ICW-P, and 74% for MBR-P, respectively, with the consistent saltwater rejection rate of 1% being observed, while R-RO exhibited an effective COD removal of 65.79%, 62.96%, and 67.72% in TW, ICW-P, and MBR-P, respectively. The highest turbidity removal of 96% in the ICW-P was also recorded for R-RO. For morphological properties, SEM analysis of the R-RO membrane revealed a likewise appearance as a UF membrane, while pore size is also comparable with the UF membrane. The most probable number (MPN) also showed complete removal of total coliforms after passing through the R-RO membrane. These features made the R-RO membrane an excellent choice for drinking water treatment and wastewater treatment polishing steps. This solution can help developing nations to be efficient in resource recovery and contribute to the circular economy.

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Vaccination/immunization plays a key role in maintaining the optimum health of humans and animals where the prevalence of disease is high and treatment becomes less effective [...].

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Plant products are widely used for health and disease management. However, besides their therapeutic effects, some plants also have potential toxic activity. Calotropis procera is a well-known laticifer plant having pharmacologically active proteins playing a therapeutically significant role in curing diseases like inflammatory disorders, respiratory diseases, infectious diseases, and cancers. The present study was aimed to investigate the antiviral activity and toxicity profile of the soluble laticifer proteins (SLPs) obtained from C. procera. Different doses of rubber free latex (RFL) and soluble laticifer protein (ranging from 0.019 to 10 mg/mL) were tested. RFL and SLPs were found to be active in a dose-dependent manner against NDV (Newcastle disease virus) in chicken embryos. Embryotoxicity, cytotoxicity, genotoxicity, and mutagenicity of RFL and SLP were examined on chicken embryos, BHK-21 cell lines, human lymphocytes, and Salmonella typhimurium, respectively. It was revealed that RFL and SLP possess embryotoxic, cytotoxic, genotoxic, and mutagenic activity at higher doses (i.e., 1.25-10 mg/mL), while low doses were found to be safe. It was also observed that SLP showed a rather safer profile as compared to RFL. This might be due to the filtration of some small molecular weight compounds at the time of purification of SLPs through a dialyzing membrane. We suggest that SLPs could be used therapeutically against viral disorders but the dose should be critically monitored.

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BACKGROUND: Intellectual disability (ID) is a neurodevelopmental condition, affecting 1-3% of the population. Genetic factors play a key role causing the limitation in intellectual functioning and adaptive behavior. The heterogeneity of ID makes it more difficult for genetic and clinical diagnosis. Mapping of variants through next generation DNA sequencing in consanguineous families would help to understand the molecular parthenogenesis of ID. OBJECTIVE: The aim of this study was to describe the genetic variants of ID in consanguineous Pakistani families. METHODS: We analyzed four unrelated consanguineous Pakistani families having an intellectual disability through whole exome sequencing (WES). Data was analyzed using different bioinformatics tools and software. RESULTS: We mapped four novel variants in different ID genes. Each variant is found in different family, co-segregating with a recessive pattern of inheritance. The variants found are; c.1437delG:p.Asn480Thrfs*10, mapped in FKRP, c.2041 C>A:p.Leu681Met in HIRA, c.382 C>T:p.Arg128Cys in BDH1 and c.267+1G>A:p.? identified in TRAPPC6B. CONCLUSIONS: These variants help in demonstration of status and molecular basis of intellectual disability in Pakistani population leading to provision of genetic counseling services and a contribution in disease variant database.

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Multi-drugs resistant (MDR) Staphylococcus aureus is creating challenges to cure cow mastitis, resulting in massive economic loss globally. It necessitates the adoption of prevention and control systems such as vaccination. Plain (PMRSAV), Montanide oil adjuvanted (MMRSAV) and Aluminum hydroxide adjuvanted (AMRSAV) vaccines were prepared using a molecularly characterized isolate of MDR S. aureus from bovine origin. Immunogenicity of the selected isolate was evaluated in five groups of rabbits (A-E) at different concentrations by measuring GMT via IHA from serum samples after booster shot. The group E provoked significantly higher (P<0.05) antibody titer with peak at day 28 (64±0.5) and cumulative mean antibody titer (CMT) of rabbits was highest (45.6) followed by groups C (35.9), D (32.7), B (30.3) and A (24.5). The concentration yielding maximum antibody titer was used for vaccines preparation. Vaccines were evaluated in different rabbits groups by inoculating PMRSAV, MMRSAV, AMRSAV and Placebo. Serum samples evaluated through IHA revealed that rabbits injected with MMRSAV produced highest antibody titer reaching its peak at day 45 (90.51±0.23) with a slight decrease until day 60 (80.63±0.17) followed by AMRSAV and PMRSAV. Challenge protection assay revealed the survival rates of rabbits in groups PMRSAV, MMRSAV, AMRSAV and Placebo as 83.3%, 100%, 100% and 16.7%, respectively. The study concluded that MMSAV and AMSAV were safe, efficacious and immunogenic in experimental rabbits.

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