Antimicrobial resistance pattern and molecular epidemiology of ESBL and MBL producing Acinetobacter baumannii isolated from hospitals in Minia, Egypt

Introduction: Multidrug resistant (MDR) Acinetobacter baumanii (A. baumannii) strains have emerged as novel nosocomial pathogens threatening patients’ lives, especially in intensive-care units (ICUs). This study aims to determine the prevalence of carbapenemase genes and CTX-M-15 and the resistance pattern of carbapenemase producing isolates. Methods: A total of 530 clinical specimens were collected from patients suffering from different infections, antibiotic susceptibility test was performed using kirby-bauer disk diffusion method. ESβL production was detected phenotypically by double-disc synergy test (DDST). Carbapenemase production was tested by Modified Hodge Test (MHT). Then, these isolates were tested for MBL detection by disc potentiation test. Carbapenemase encoding genes (VIM, IMP, GIM and SPM, OXA-51, OXA-23 and OXA-143) and CTX-M-15 were tested by polymerase chain reaction (PCR). Results: Out of 530 samples, 20 bacterial isolates were identified as A. baumannii from different infectious cases, 35% of isolates were ESBL-producers. Eleven isolates were resistant to imipenem (4 isolates) and meropenem (7 isolates). All carbapenem resistant isolates were MHT positive. Nine (45%) isolates were confirmed as A. baumannii by OXA-51 (all were carbapenem resistant). Distribution of IMP, VIM, GIM and SPM, OXA-23, OXA-143 and CTX-M-15 by PCR were 55, 50, 50, 25, 35, 45 and 33% respectively. Conclusion: The high prevalence of resistance genes and the resistance pattern of the isolates indicate that the detection of ESBLs and MBLs phenotypically and genotypically with the study of the resistance pattern of the isolates is critically important for the surveillance of drug resistance in the hospital environment

In vitro inhibition of uropathogenic Escherichia coli biofilm formation by probiotic Lactobacilli isolated from healthy breast fed infants

Biofilm forming Escherichia coli bacterium exhibits multiple drug resistance, which is responsible for recurrent urinary tract infections (UTI) that are difficult to eradicate. The work aimed to investigate the antibacterial and antibiofilm activities of Lactobacilli isolated from faecal microbiota of healthy infants; to identify these isolates and determine their probiotic characteristics. E. coli isolates were recovered from urine samples of patients with urinary tract infections. On the other hand, Lactobacillus isolates were recovered from faeces of breast-fed infants. Five strong biofilm forming E. coli isolates with multidrug resistance (MDR) were selected. The antibacterial potential of Lactobacillus supernatants were assessed via disk diffusion assay. All the tested E. coli isolates showed high susceptibility to the Lactobacillus supernatants; where 54 % of these supernatants expressed inhibition zones diameters ranging from 15- 18 mm. Antibiofilm efficacies of Lactobacillus spp. against E. coli isolates were tested in vitro using microtiter plate assay and scanning electron microscopy (SEM). More than 50 % reduction of biofilms formation by the 5 selected MDR E. coli isolates was observed by most the Lactobacillus isolates. The scanning electron microscopy confirmed the elimination of E. coli biofilms by cells of the Lactobacillus isolates. Preliminary probiotic characteristics of the Lactobacillus isolates were investigated; all isolates tolerated 2 % bile salt concentration and acidic condition at pH 3. Regarding safety of the Lactobacilli for human consumption, all isolates were non hemolytic, and 14 Lactobacillus isolates were sensitive to all tested antibiotics except for vancomycin, as they are naturally resistant to it. About 14 safe probiotic Lactobacillus isolates were identified by API-50 CHL test as; Lactobacillus acidophilus, L. plantarum, L. fermentum and L. paracasei

Extracellular biosynthesis of silver nanoparticles using Bacillus subtilis and their antibacterial activity against clinical bacterial species

The aims of this study were to biosynthesize silver nanoparticles (AgNPs) using Bacillus subtilis supernatant, and to evaluate their in vitro antibacterial potential against human pathogens; namely Staphylococcus aureus (Staph. aureus) and Escherichia coli (E. coli). Nanoparticles (NPs) are becoming popular in different fields of research, and are useful in combating vast number of microbial diseases. NPs may be artificially synthesized in vitro using chemical methods and\or via extracellular metabolites produced by the bacterial strains. In the present study, biosynthesis of AgNPs was carried out in vitro using supernatants of B. subtilis. Biosynthesized AgNPs were characterized through several physical methods. The recorded Z-average (d. nm) was 135.0 nm; with 99.2 % of the NPs displaying a hydrodynamic distance across of 188.0 nm (SD= 117.7). The polydispersity index was 0.246 and the Zetapotential value was - 17.2 mV, which indicates good colloidal stability. Results of the Transmission electron microscope (TEM) observation indicated that the particles were spherical in shape with an average size of 21.8- 27.5 nm. The antibacterial efficacy of the AgNPs against Methicillin resistant Staph. aureus (MRSA) and E. coli clinical isolates was evaluated in vitro using the agar well diffusion. The AgNPs demonstrated antibacterial potential against MRSA and E. coli isolates; recording 18 and 15 mm diameter of zones of inhibition, respectively. The minimum inhibitory concentration (MIC) was found to be 142 µg/ ml, while the recorded minimum bactericidal concentration (MBC) was 284 µg/ ml. The mode of action of the AgNPs was investigated using the Scanning electron microscope (SEM), which was recognized as bacterial cell lysis and elongation. Current data suggest an efficient biosynthesis of stable AgNPs by B. subtilis with remarkable antibacterial potential

Efficacy and durability of bovine virus diarrhea (BVD) virus killed vaccine adjuvanted with monolaurin.

The bovine virus diarrhea virus (BVDV) causes reproductive, enteric, and respiratory diseases. Vaccination is essential in increasing herd resistance to BVDV spread. The selection of an adjuvant is an important factor in the success of the vaccination process. Monolaurin or glycerol monolaurate is a safe compound with an immunomodulatory effect. This study aimed to evaluate the efficacy of monolaurin as a novel adjuvant. This was examined through the preparation of an inactivated BVDV (NADL strain) vaccine adjuvanted with different concentrations of monolaurin and compared with the registered available locally prepared polyvalent vaccine (Pneumo-4) containing BVD (NADL strain), BoHV-1 (Abou Hammad strain), BPI3 (strain 45), and BRSV (strain 375L), and adjuvanted with aluminum hydroxide gel. The inactivated BVDV vaccine was prepared using three concentrations, 0.5%, 1%, and 2%, from monolaurin as adjuvants. A potency test was performed on five groups of animals. The first group, which did not receive vaccination, served as a control group while three other groups were vaccinated using the prepared vaccines. The fifth group received the Pneumo-4 vaccine. Vaccination response was monitored by measuring viral neutralizing antibodies using enzyme-linked immunosorbent assay (ELISA). It was found that the BVD inactivated vaccine with 1% and 2% monolaurin elicited higher neutralizing antibodies that have longer-lasting effects (nine months) with no reaction at the injection site in comparison to the commercial vaccine adjuvanted by aluminum hydroxide gel

The Inc FII Plasmid and its Contribution in the Transmission of blaNDM-1 and blaKPC-2 in Klebsiella pneumoniae in Egypt

The emergence of blaKPC-2 and blaNDM-1 producing Klebsiella pneumoniae represents a great problem in many Egyptian hospitals. One hundred and twenty-six K. pneumoniae isolates from patients admitted to Assiut University Hospital were identified by an API20E kit. Carbapenemase-producing K. pneumoniae (CPKP) was detected by the modified carbapenem inactivation method (mCIM), the EDTA-modified carbapenem inactivation method (eCIM), and an E-test. Based on the polymerase chain reaction, all isolates were negative for bla-VIM-1 and bla-IMP-1, fifteen of these isolates were positive for both blaKPC-2 and blaNDM-1, two isolates were positive for blaKPC-2 only, and twenty-eight isolates were positive for bla-NDM-1 only. Although one isolate was positive for the string test, all CPKP isolates were negative for capsular genes. Only 71.1% of CPKP transferred their plasmids to their corresponding transconjugants (E. coli J53). The resistance patterns of the clinical isolates and their transconjugates were similar, except for 12 isolates, which showed differences with their transconjugates in the resistance profile of four antibiotics. Molecular typing of the plasmids based on replicon typing showed that Inc FIIK and FII plasmids predominated in isolates and their transconjugants carrying blaKPC-2 and/or blaNDM-1. Conjugative Inc FII plasmids play an important role in the spread of CPKP, and their recognition is essential to limit their spread

Association of Hepatitis C viral load with liver functions and risk factors among HCV patients, Minia governorate, Egypt

Hepatitis C virus (HCV) is one of the blood transmitted hepatitis viruses. HCV infections have been identified as major causes of chronic hepatic diseases, and hepatocellular carcinoma. The aims of the current study were to determine the HCV viral load between 35 hepatitis C patients in Minia governorate, Egypt, and to assess association of the viral load with abnormal liver functions including; Alanine aminotransferase (ALT), prothrombin activity and platelet count. In addition to assessing if there are any risk factors associated with the population group, sex, age and other factors. About 35 blood samples were collected from hepatitis C patients randomly selected from the outpatient clinic at the Viral Hepatitis Management Center, Minia governorate, Egypt; including males and females of different ages. Viral load was determined using Real-time polymerase chain reaction (RT-PCR). All relevant information was collected from each patient including personal and clinical data. Current results showed that 68.60 % of the samples were from males and 31.4 % were from females, and most of them aged between 51 and 70 years. Approximately 11 (31.4 %) of the HCV patients had viral loads of <106, 12 (34.3 %) recorded viral loads of <105, and about 12 cases (34.3 %) had a viral load of < 104. HCV infection has been associated with 4 risk factors representing high HCV transmission routes including; dental intervention (80.0 %), history of hospital admission (65.7 %), previous surgeries (57.1 %) and family history of HCV (48.6 %). However, history of Schistosomiasis and blood-transfusion showed low association with HCV infection; recording (31.4 %) and (22.9 %), respectively

Influence of selected non-antibiotic pharmaceuticals on antibiotic resistance gene transfer in Escherichia coli.

BackgroundAntibiotic resistance genes (ARGs) transfer rapidly among bacterial species all over the world contributing to the aggravation of antibiotic resistance crisis. Antibiotics at sub-inhibitory concentration induce horizontal gene transfer (HRT) between bacteria, especially through conjugation. The role of common non-antibiotic pharmaceuticals in the market in disseminating antibiotic resistance is not well studied.ObjectivesIn this work, we indicated the effect of some commonly used non-antibiotic pharmaceuticals including antiemetic (metoclopramide HCl) and antispasmodics (hyoscine butyl bromide and tiemonium methyl sulfate) on the plasmid-mediated conjugal transfer of antibiotic resistance genes between pathogenic E. coli in the gastric intestinal tract (GIT).MethodsBroth microdilution assay was used to test the antibacterial activity of the tested non-antibiotic pharmaceuticals. A conjugation mating system was applied in presence of the studied non-antibiotic pharmaceuticals to test their effect on conjugal transfer frequency. Plasmid extraction and PCR were performed to confirm the conjugation process. Transmission electron microscopy (TEM) was used for imaging the effect of non-antibiotic pharmaceuticals on bacterial cells.ResultsNo antibacterial activity was reported for the used non-antibiotic pharmaceuticals. Plasmid-mediated conjugal transfer between isolates was induced by metoclopramide HCl but suppressed by hyoscine butyl bromide. Tiemonium methylsulfate slightly promoted conjugal transfer. Aggregation between cells and periplasmic bridges was clear in the case of metoclopramide HCl while in presence of hyoscine butyl bromide little affinity was observed.ConclusionThis study indicates the contribution of non-antibiotic pharmaceuticals to the dissemination and evolution of antibiotic resistance at the community level. Metoclopramide HCl showed an important role in the spread of antibiotic resistance

Design, synthesis and molecular docking of new N-4-piperazinyl ciprofloxacin-triazole hybrids with potential antimicrobial activity

New N-4-piperazinyl ciprofloxacin-triazole hybrids 6a-o were prepared and characterized. The in vitro antimycobacterial activity revealed that compound 6a experienced promising antimycobacterial activity against Mycobactrium smegmatis compared with the reference isoniazide (INH). Additionally, compound 6a exhibited broad spectrum antibacterial activity against all the tested strains either Gram-positive or Gram-negative bacteria compared with the reference ciprofloxacin. Also, compounds 6g and 6i displayed considerable antifungal activity compared with the reference ketoconazole. DNA cleavage assay of the highly active compounds 6c and 6h showed a good correlation between the Mycobactrium cleaved DNA gyrase assay and their in vitro antimycobactrial activity. Moreover, molecular modeling studies were done for the designed ciprofloxacin derivatives to predict their binding modes towards Topoisomerase II enzyme (PDB: 5bs8)

COVID-19 associated Mucormycosis among ICU patients: risk factors, control, and challenges

Abstract The severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) pandemic is still difficult to be controlled. The spread of this virus and the emergence of new variants are considered a great challenge worldwide. Disturbance in infection control guidelines implementation, use of steroids, antibiotics, hospital crowdedness, and repeated use of oxygen masks during the management of critically ill COVID-19 patients lead to an increase in the rate of opportunistic infections. So, patients need to fight both the virus with its different variants and opportunistic pathogens including bacteria and fungi especially patients with diabetes mellitus, malignancy, or those who undergo hemodialysis and receive deferoxamine. During the pandemic, many cases of Mucormycosis associated with COVID-19 infection were observed in many countries. In this review, we discuss risk factors that increase the chance of infection by opportunistic pathogens, especially fungal pathogens, recent challenges, and control measures

Global economic burden of unmet surgical need for appendicitis

Background There is a substantial gap in provision of adequate surgical care in many low- and middle-income countries. This study aimed to identify the economic burden of unmet surgical need for the common condition of appendicitis. Methods Data on the incidence of appendicitis from 170 countries and two different approaches were used to estimate numbers of patients who do not receive surgery: as a fixed proportion of the total unmet surgical need per country (approach 1); and based on country income status (approach 2). Indirect costs with current levels of access and local quality, and those if quality were at the standards of high-income countries, were estimated. A human capital approach was applied, focusing on the economic burden resulting from premature death and absenteeism. Results Excess mortality was 4185 per 100 000 cases of appendicitis using approach 1 and 3448 per 100 000 using approach 2. The economic burden of continuing current levels of access and local quality was US $92 492 million using approach 1 and$73 141 million using approach 2. The economic burden of not providing surgical care to the standards of high-income countries was $95 004 million using approach 1 and$75 666 million using approach 2. The largest share of these costs resulted from premature death (97.7 per cent) and lack of access (97.0 per cent) in contrast to lack of quality. Conclusion For a comparatively non-complex emergency condition such as appendicitis, increasing access to care should be prioritized. Although improving quality of care should not be neglected, increasing provision of care at current standards could reduce societal costs substantially