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Bilal Aslam,1 Muhammad Hussnain Siddique,2 Abu Baker Siddique,1 Muhammad Shafique,1 Saima Muzammil,1 Mohsin Khurshid,1 Muhammad Hidayat Rasool,1 Moeed Ahmad,1 Tamoor Hamid Chaudhry,3 Afreenish Amir,3 Muhammad Salman,3 Zulqarnain Baloch,4 Norah A Alturki,5 Ahmad Alzamami6 1Department of Microbiology, Government College University Faisalabad, Faisalabad, Pakistan; 2Department of Bioinformatics & Biotechnology, Government College University Faisalabad, Faisalabad, Pakistan; 3Public Health Laboratories Division, National Institute of Health, Islamabad, Pakistan; 4Faculty of Life Science and Technology, Kunming University of Science and Technology, Kunming, People’s Republic of China; 5Clinical Laboratory Science Department, College of Applied Medical Science, King Saud University, Riyadh, Saudi Arabia; 6Clinical Laboratory Science Department, College of Applied Medical Science, Shaqra University, AlQuwayiyah, Saudi ArabiaCorrespondence: Bilal Aslam, Department of Microbiology, Government College University Faisalabad, Faisalabad, Pakistan, Email drbilalaslam@gcuf.edu.pk Ahmad Alzamami, Clinical Laboratory Science Department, College of Applied Medical Science, Shaqra University, AlQuwayiyah, Saudi Arabia, Email aalzamami@su.edu.saBackground: The World Health Organization (WHO) has declared the multi-drug resistant (MDR) Klebsiella pneumoniae as one of the critical bacterial pathogens. The dearth of new antibiotics and inadequate therapeutic options necessitate finding alternative options. Bacteriophages are known as enemies of bacteria and are well-recognized to fight MDR pathogens.Methods: A total of 150 samples were collected from different clinical specimens through a convenient sampling technique. Isolation, identification, and antibiotic susceptibility testing (AST) of K. pneumoniae were done by standard and validated microbiological procedures. Molecular identification of virulence factors and antibiotic resistance genes (ARGs) was carried out through polymerase chain reaction (PCR) by using specific primers. For bacteriophage isolation, hospital sewage samples were processed for phage enrichment, purification, and further characterization ie, transmission electron microscopy (TEM) and stability testing, etc. followed by evaluation of the lytic potential of the phage.Results: Overall, a total of 41% of isolates of K. pneumoniae were observed as hypervirulent K. pneumoniae (hvKp). Among hvKp, a total of 12 (42%) were detected as MDR hvKp. A total of 37% of all MDR isolates were found resistant to colistin, and 66% of the colistin resistance isolates were recorded as mcr-1 positive. Isolated phage KpnM had shown lytic activity against 53 (79%) K. pneumoniae isolates. Remarkably, all 8 mcr-1 harboring MDR hvKp and non-hvKp isolates were susceptible to KpnM phage.Conclusion: Significant distribution of mcr-1 harboring hypervirulent Klebsiella pneumoniae was observed in clinical specimens, which is worrisome for the health system of the country. Characterized phage KpnM exhibited encouraging results and showed the lytic activity against the mcr-1 harboring hvKp isolates, which may be used as a prospective alternative control strategy to fight this ominous bacterium.Keywords: Klebsiella pneumoniae, bacteriophages, colistin, MDR, virulence

The local climate of cities is changing, and one of the primary reasons for this change is rapid urbanization. The Lahore district is situated in the Punjab province of Pakistan and is mainly compr...

Introduction Intense livestock farming practices enforcing the farmers to use antibiotics as food supplements on a routine basis. Aberrant use of antibiotics is associated with the emergence of antibiotics resistance and resistant superbugs. Keeping in view the current scenario, the present study was designed for the first time from Pakistan with a specific aim to estimate the prevalence of the carbapenem-resistant Klebsiella pneumoniae in veterinary settings and the waste in Pakistan. Methods A total of 138 samples from various veterinary sources were collected by employing a nonprobability sampling technique. Isolation and phenotypic identification of carbapenem-resistant K. pneumoniae were performed according to the CLSI standard. Molecular detection of various antibiotic resistance genes (ARGs) was done through PCR by using specific primers against each ARG. According to the pasture scheme, the multilocus sequence typing (MLST) was performed to characterize the K. pneumoniae sequence types (STs). Results According to the results of the study, overall 9.4% (13/138) isolates were confirmed carbapenem-resistant K. pneumoniae. Among various carbapenem ARGs particularly, the bla NDM-1 was found in 92.3% (12/13) isolates followed by bla OXA-48 84.6% (11/13). MLST results revealed that overall 3 STs were found in the study which includes ST29, ST11, and ST258. Taking together, this is the first study to our best knowledge which demonstrated the prevalence of carbapenem-resistant K. pneumoniae and its various STs prevalent in veterinary settings and the waste of Pakistan. Conclusion Based on the above-mentioned facts, we suggested that veterinary settings and waste are the potential source and reservoir of carbapenem-resistant K. pneumoniae, which may be disseminated to the environment and ultimately can affect the public and companion livestock health.

Introduction The aminoglycosides are widely used for the therapeutic management of infections caused by gram-negative bacteria, including the Acinetobacter baumannii strains. However, the resistance to the members of the aminoglycoside family, such as amikacin, gentamicin, and tobramycin, is increasingly being common among the clinical isolates. Purpose This study aimed to investigate the presence of 16SrRNA methylases and aminoglycoside modifying enzymes (AMEs) genes among aminoglycoside resistant A. baumannii isolates and to study the genetic diversity of the clinical population of A. baumannii in local hospitals. Material and methods The 143 A. baumannii clinical strains were analyzed for antimicrobial susceptibility, genetic screening for enzymes conferring aminoglycosides resistance followed by the multilocus sequence typing. Results The 133/143 (93%) isolates were non-susceptible to at least one of the tested aminoglycosides, including amikacin, gentamicin, and tobramycin. The MIC distribution has shown that 87.486.7% strains were resistant to amikacin and gentamicin, respectively. The aphA6, aadB, aacC1, and aphA1 were found in 74.1%, 59.4%, 16.1%, and 11.2% isolates, respectively, whereas the armA was found in 28% of the strains having a higher MIC value (MIC; ≥256µg/mL). The MLST data have shown that the ST589 and ST2 were the most common STs and corresponded to 51 (35.7%) and 38 (26.6%) isolates, respectively, and few of the isolates corresponding to these STs were found to harbor the armA gene with a variable genotypic profile for AMEs. Discussion The study has reported the incidence of various enzymes conferring aminoglycoside resistance among the A. baumannii clones for the first time from Pakistan. The findings suggest the possibility of transmission of aminoglycoside resistance determinants through the lateral gene transfer as well as clonal dissemination.

Objective To assess the association between triglycerides (TG), total cholesterol (TC) and hyperuricemia (HUA) in the general Chinese population. Methods A population-based cross-sectional survey included 9680 participants aged 35-74 years in 2006 and 2009 in Qingdao, China. TG, TC and uric acid (UA) were measured. The logistic regression model was performed to estimate the association between TG, TC, and HUA with an odds ratio (OR) and 95% confidence intervals (CI). Meanwhile, age stratification analysis (

The New Delhi Metallo-β-lactamase (NDM) producing Enterobacteriaceae is spreading worldwide. Although the blaNDM gene has been identified in animal associated Enterobacteriaceae isolates in many countries, little is known about its occurrence in animal products in Pakistan. In this study, 13 Escherichia coli isolates were collected from chicken meat samples in Pakistan. Two isolates, 15978 and C4109, exhibited reduced susceptibility (MIC ≥1 μg/mL) to imipenem, and carried blaNDM-5 and blaNDM-7 gene, respectively. Whole-genome sequencing and Oxford Nanopore MinION sequencing revealed that 15978 and C4109 belonged to ST156 and ST167, respectively. blaNDM-7 was carried by an IncX3 plasmid that has disseminated worldwide, whereas blaNDM-5 was located on an F36: A-: B32 plasmid, which shared high identity with two plasmids carried by E. coli isolates from other countries (one from a patient in Canada). To the best of our knowledge, this is the first report characterizing blaNDM-carrying plasmids from chicken meat samples in Pakistan. The dissemination of almost identical blaNDM-5-bearing F36:A-:B32 and blaNDM-7-bearing IncX3 plasmids in different countries highlights the importance of international trade and travel in the spread of antimicrobial resistance strains and plasmids worldwide.

RFID tag antennas with stable performance on the diverse electromagnetic mounting platforms are an integral part of the ubiquitous RFID systems. This research article presents a novel tag antenna d...

The advent of multidrug resistance among pathogenic bacteria is imperiling the worth of antibiotics, which have previously transformed medical sciences. The crisis of antimicrobial resistance has been ascribed to the misuse of these agents and due to unavailability of newer drugs attributable to exigent regulatory requirements and reduced financial inducements. Comprehensive efforts are needed to minimize the pace of resistance by studying emergent microorganisms, resistance mechanisms, and antimicrobial agents. Multidisciplinary approaches are required across health care settings as well as environment and agriculture sectors. Progressive alternate approaches including probiotics, antibodies, and vaccines have shown promising results in trials that suggest the role of these alternatives as preventive or adjunct therapies in future.

Zulqarnain Baloch,1,* Bilal Aslam,2,* Saima Muzammil,2 Mohsin Khurshid,3 Muhammad Hidayat Rasool,2 Ke Ma4 1College of Veterinary Medicine, South China Agricultural University, Guangzhou, China; 2Department of Microbiology, Government College University Faisalabad, Faisalabad, Pakistan; 3College of Allied Health Professionals, Directorate of Medical Sciences, Government College University Faisalabad, Faisalabad, Pakistan; 4College of Traditional Chinese Medicine, Shandong University of Traditional Chinese Medicine, Jinan, China *These authors contributed equally to this work Abstract: Antibiotic therapy has a dual impact: wanted, in which it immediately inhibits the growth of bacteria and the unwanted, which is responsible for the evolution of antibiotic resistance. The dissociation of therapeutic effectiveness from the possible risk of the antibiotic resistance may be attained by taking the advantage of specific relations between these drugs, and the methods in which mutations associated with resistance against a specific antibiotic may modify these relations or it may increase the sensitivity of the bacterium to the other antibiotics. Although the practical implementation of this notion needs considerable advancement and confirmation that depends upon the improvements in the field of genomics and diagnostics, these interventions propose new paradigms, which may confine or inverse the evolution of antibiotic resistance. Keywords: antibiotic resistance, inversion, mutations, bacteria, evolution

The broad-spectrum antibacterial drugs are widely used in food producing animals. Suspected residues of these drugs in meat may have ill effects upon human health. The aim of the present study was to determine the enrofloxacin residues in broiler’s meat and liver samples. Detection of enrofloxacin residues in meat (n = 75) and liver (n = 75) samples was performed by high performance liquid chromatography with UV detector set at 268 nm using C18 column. Overall mean residual concentrations of enrofloxacin in meat and liver samples were 208 ± 55 and 527 ± 84 µg/kg, respectively. This study revealed that 52% (39) meat and 78.7% (59) liver samples were positive for enrofloxacin, out of these 58.3% (21) meat and 71.2% (42) liver samples were having residual concentration above the maximum residual limits. So it can be concluded that the usage of this contaminated meat may cause resistance in consumers and seems to be a public health threat.

Infectious diseases are major constraint that hinders the poultry industry. Among them parasitic diseases are very common and Ascaridia galli is one of the most common parasitic roundworms found in poultry. Haemorrhages, diarrhoea and listlessness are signs of infection. Parasitic infections such as A. galli are treated with chemical anthelmintics (piperazine, albendazole, levamisole, Ivermectin, benzimidazoles and fenbendazole). These synthetic chemicals can promote resistance, so there is need for alternative ways to treat the disease. Medicinal plants have the potential to combat such parasitism and the development of anthelmintic resistance appears to be very slow against such treatment. This review covers the studies related to the screening of plant materials having in vitro and in vivo anthelmintic activities against A. galli throughout the world. Medicinal plants showing in vitro anthelmintic activity include Anacardium occidentale, Allium sativum, Tribulus terrestris, Bassia latifolia, Piper betle, Morinda citrifolia L.I, Cassia occidentalis L. and Aloe secundiflora while in vivo studies include the use of Psorelia corylifolia, Piper betle, Pilostigma thonningi, Caesalpinia crista, Ocimum gratissimum and Anacardium occidentale. In conclusion, medicinal plants appear to have good anthelmintic activities in poultry and may substitute conventionally used synthetic drugs, and their use may moderate drug resistance in endemic pathogen populations and drug residues in poultry meat.

The poultry industry is affected by economically important problems such as Newcastle disease (ND). Newcastle disease virus (NDV) belongs to the Paramyxoviridae family and manifests as different strains; lentogenic, mesogenic and velogenic. ND affects the quality of eggs and impairs growth performance of birds. Various efforts have been made to control economic losses due to this disease, including using live and killed vaccines, which do not confer 100% immunity in all cases. Due to mutations within viral strains, NDV can become resistant and difficult to control, and there is a need to search for alternative measures. Medicinal plants are considered as a complementary means to control this virus, especially in developing countries, because they have been implicated in treatments for a variety of infectious and non-infectious diseases. Alkaloids, flavonoids, saponins and tannins found in plants have been regarded as novel antiviral agents. The following paper encompasses the studies related to plants having in vitro antiviral activity against ND such as Momordica balamina, Adansonia digitata, Artemisia annua, Azadirachta indica, Psidium guajava, Moringa oleifera and plants having good in vivo antiviral activity such as Aloe secundiflora, Cucumis metuliferus and Anthocleista nobilis as well. It discusses plants with the potential to improve the immunity of infected birds, such as Mangrove halophytes, Nigella sativa, Polysavone, Melissa officinalis, Momordica cochinchinensis, Echinacea purpurea, Withania somnifera and Aspargus Racemosus.

The potential health risk due to lifetime exposure to copper, lead, chromium, zinc, and iron in urban soil of Karachi, Pakistan, was evaluated. Mean concentrations of Cu, Pb, Cr, Zn, and Fe in topsoil samples were 33.3 ± 12.8, 42.1 ± 55.8, 9.6 ± 4.2, 99.5 ± 37.3, and 908.4 ± 57.8 mg kg−1, respectively. A U.S. Environmental Protection Agency model was adopted for the carcinogenic and non-carcinogenic risk assessment from different exposure pathways. Risk assessment indicated that the overall results for the carcinogenic risk were insignificant. However, the carcinogenic risk from Pb due to oral ingestion of soil exceeded the value of 1 × 10−6, in some areas of the city. It indicates that the exposure to Pb-contaminated soil may cause adverse health effects in humans, especially in children. The Hazard Quotient (HQ) for different metals through ingestion and dermal pathways was also found to be less than 1. The combined Hazard Index (HI) for children through different routes of exposure was 8.9 tim...