Pseudomonas aeruginosa is considered as foremost cause of hospital acquired infections due to its innate and plasmid mediated resistance to multiple antibiotics making it a multi drug resistant (MDR) pathogen. Biofilm formation is a pathogenic mechanism harbored by this pathogen which further elevates its resistance to antibiotics and host defense system. The aim of the present study was to evaluate the biofilm forming potential and distribution of pslA gene in multi drug resistant Pseudomonas aeruginosa isolates obtained from different clinical samples. A total of 200 different clinical samples were collected after obtaining written consent from the patients. The samples were subjected to isolation and identification of P. aeruginosa by standard microbiological procedures. Confirmation of isolates was done by polymerase chain reaction targeting oprL gene. Kirby Bauer method was performed for detection of MDR isolates. Congo red agar (CRA) test and Microtiter plate assay (MPA) for observing the biofilm forming ability and amplification of pslA gene was also performed on MDR isolates. The results showed that from 200 samples 52 (26 %) were P. aeruginosa and among them 20 (38.46 %) were MDR isolates. The CRA showed 23 (44.23 %) while MPA detected 49 (94.23 %) isolates as biofilm producers while all the MDR isolates showed biofilm formation by MPA method. The pslA gene was detected in all biofilm forming isolates while 90 % in MDR P. aeruginosa. It was concluded that biofilm forming P. aeruginosa are more resistant to tested antibiotics and biofilm formation is strongly associated with presence of pslA gene.

Background: Human brucellosis is a neglected and zoonotic disease also known as Malta fever or undulant fever. Objectives: A cross sectional study was conducted to determine the seroprevalence of brucellosis and to access the role of risk factors associated with this disease in a sample from Punjab, Pakistan. Methods: A total of 250 serum samples were collected and subjected to Rose Bengal Plate Test and Enzyme Linked Immunosorbent Assay for screening of Brucella. A predesigned questionnaire was filled prior to sampling to collect data regarding socio - demographic and suspected risk factors of human brucellosis. Descriptive and bivariate statistical analysis was performed using the STATA software version 12. Results: The study showed 16% seroprevalence of brucellosis. The prevalence was statistically higher in males (24%), age group of 20 to 30 years (26.92%), rural residents, (23%) and individuals with animals at home (22.50%). Among the related risk factors, exposure to animals (OR = 1.87, 95% CI: 0.9459, 3.6973) and consuming raw milk (OR = 2.36, 95% CI: 1.1713, 4.7760) were strongly associated with the disease. The results showed that white blood cells were significantly (P < 0.05) lowered in Brucella infected individuals. Conclusions: Awareness programs in the rural population should be provided about the disease and its associated risk factors. Consuming unpasteurized milk and products should be avoided to control this disease. Keywords: Brucella; Seroprevalence; Risk Factors; Haematological Parameters; Pakistan

BACKGROUND:The prevalence of Giardia lamblia in Pakistani children is currently unknown.The aim here was to evaluate the prevalence and risk factors of Giardia lamblia in children exhibiting diarrhea. DESIGN AND SETTING: Cross-sectional study at different district healthcare hospitals in Pakistan. METHODS: A total of 800 samples were collected from children aged 0-10 years. Information regarding personal data, demographic data and supposed risk factors was collected through a structured questionnaire. Giardia lamblia was detected through direct microscopy and antigens through the enzyme-linked immunosorbent assay (ELISA). RESULTS: The prevalence of Giardia lamblia was 2.75% through direct microscopy and inflated to 9.5% through ELISA. The demographic factors positively associated with occurrences of giardiasis were age (P = 0.035; odds ratio, OR = 1.96; 95% confidence interval, CI = 1.094-3.533), mother's educational level (P = 0.031; OR = 2.67; 95% CI = 1.186-6.045) and father's educational level (P = 0.004; OR = 3.56; 95% CI = 1.612-7.899). Similarly, among the supposed risk factors, rural residency (P = 0.032; OR = 1.76; 95% CI = 1.098- 2.851), absence of proper sewerage system (P = 0.000; OR = 6.60; 95% CI = 4.029-10.841) and unavailability of safe drinking water (P = 0.000; OR = 4.08; 95% CI = 2.207-7.547) were the factors strongly connected with giardiasis. Abdominal discomfort was a prominent clinical sign with 46% frequency. CONCLUSION: Various risk factors were associated with occurrences of Giardia, thus emphasizing the importance of parent's education, safe drinking water and proper sewerage systems for Pakistani children's health.

The emergence of carbapenem-resistant bacterial pathogens is a significant and mounting health concern across the globe. At present, carbapenem resistance (CR) is considered as one of the most concerning resistance mechanisms and mainly found in gram-negative bacteria of the Enterobacteriaceae family. Although carbapenem resistance has been recognized in Enterobacteriaceae from last 20 years or so, recently it emerged as a global health issue as CR clonal dissemination of various Enterobacteriaceae members especially E. coli, and Klebsiella pneumoniae are reported from across the globe at an alarming rate. Phenotypically, carbapenems resistance is in due to the two key mechanisms, like structural mutation coupled with β-lactamase production and the ability of the pathogen to produce carbapenemases which ultimately hydrolyze the carbapenem. Additionally, penicillin-binding protein modification and efflux pumps are also responsible for the development of carbapenem resistance. Carbapenemases are classified into different classes which include Ambler classes A, B, and D. Several mobile genetic elements (MGEs) have their potential role in carbapenem resistance like Tn4401, Class I integrons, IncFIIK2, IncF1A, and IncI2. Taking together, resistance against carbapenems is continuously evolving and posing a significant health threat to the community. Variable mechanisms that are associated with carbapenem resistance, different MGEs, and supplementary mechanisms of antibiotic resistance in association with virulence factors are expanding day by day. Timely demonstration of this global health concern by using molecular tools, epidemiological investigations, and screening may permit the suitable measures to control this public health menace.

Background. The emergence of resistance to beta‐lactam agents in poultry results in multidrug‐resistant (MDR) phenotypes in Escherichia coli isolates from poultry birds. The appearance of mobile colistin resistance (mcr) genes in the poultry sector has further worsened the situation. Therefore, the current study is aimed at investigating the molecular epidemiology of mcr harboring colistin‐resistant E. coli among poultry. Methods. The isolation and identification of colistin‐resistant E. coli (CR‐Ec) were done from the broiler’s fecal samples through culturing using selective media supplemented with colistin sulfate (4 μg/ml). The antibiogram studies of the isolates were performed using the disc diffusion method and broth microdilution method as per CLSI guidelines. The screening for the genes conferring resistance to colistin as well as beta‐lactam agents was performed by PCR. The genetic diversity of mcr-positive strains was assessed by multilocus sequencing typing (MLST). Results. Out of 500 fecal samples, 7% (35/500) were found positive for the presence of colistin‐resistant E. coli (CR‐Ec). Among the CR‐Ec isolates, 74.28% (26/35) were detected as ESBL producers carrying the blaCTX‐M‐1 gene in 15/35 (42.85%) isolates and blaCTX‐M‐15 and blaTEM genes in 21/35 (60%) and 35/35 (100%) isolates, respectively. E. coli isolates were found positive for the presence of mcr‐1, although none of the isolates exhibited the mcr‐2 or mcr‐3 genes. The MLST of CR‐Ec has shown the ST1035 as the most prevalent genotype, while 82.85% (29/35) of CR‐Ec strains belonged to clonal complex (CC) 131 comprising ST1035, ST131, ST1215, ST1650, and ST2279. Conclusions. The findings suggest a continuous monitoring system in veterinary and clinical settings to avoid unnecessary antibiotics. Further studies are needed at the national level to help control the increasing resistance among Enterobacterales in poultry settings.

Purpose: – The purpose of this paper is to evaluate the frequency and antimicrobial susceptibility pattern of pathogens present in ready-to-eat salads available at a local market. Design/methodology/approach: – A 100 salad samples were collected aseptically. Each sample (25 g) was homogenized in 225 ml of sterile peptone water and was serially diluted up to 1×106. Dilutions were inoculated on nutrient agar by surface spread plate technique. Aerobic colony count (ACC) was estimated by counting the colonies. Bacterial isolates were cultured on blood and MacConkey agar and identified on the basis of their morphology, culture characteristics and confirmed by API 20E and 20NE. Antimicrobial susceptibility was determined as per CLSI 2014. Findings: – ACC range was 1.1×103 cfu/g to 5.8×109 cfu/g. Among these the highest ACC was found in channa chat (4.9×104 to 5.8×109 cfu/g). A total of 127 microorganisms were identified; 73 were gram negative rods (GNRs) and 24 were gram positive cocci (GPC). Among GNRs; Klebsiella spp. (n=18) was the predominant whereas among GPC, Staphylococcus aureus (n=6) were the chief pathogen. Klebsiella spp. showed 100 percent resistance to ampicillin, 89-78 percent to amoxicillin/clavulanic acid and 33 percent to imipenem, however, Enterobacter spp. were resistant to ampicillin (100 percent) amoxicillin/clavulanic acid (77 percent) and imipenem (23 percent). Staphylococcus aureus showed resistance to co-amoxiclav (83 percent) and penicillin (75 percent). Practical implications: – This study revealed that effective control measures must been implemented and respected by throughout the entire food chain and better surveillance studies should be performed at national level to reduce the spread of bacteria by fresh salads. Originality/value: – This paper explore the high prevalence of multidrug-resistant pathogens in different salads and most of the salads were found to be unhygienic for consumption.