Diversity of biofilm-specific antimicrobial resistance genes in Pseudomonas aeruginosa recovered from various clinical isolates.

Background and Objectives: The resistance of Pseudomonas aeruginosa to antibiotics offers a significant challenge in the treatment of patients. This study aimed to investigate the antimicrobial resistance profile, biofilm-specific antimicrobial resistance genes, and genetic diversity of P. aeruginosa recovered from clinical samples. Materials and Methods: Totally 47 non-duplicate isolates of P. aeruginosa were recovered from various clinical samples. toxA, algD, ndvB, and tssC1 genes were detected in biofilm-producing isolates. The DNA sequences of the toxA and tssC1 genes were analyzed, by creating phylogenetic trees. Results: The findings revealed that 30 (63.8%) of the isolates tested positive for Extended spectrum ß-lactamase (ESBL), whereas 31 (65.9%) tested positive for Metallo-ß-lactamase (MBL) and all of the isolates presented the toxA genes, and 19.1%,17%, 6.3% presented by algD, ndvB and tssC1 genes. Besides, the phylogenetic trees of the toxA and tssC1 gene isolates suggested a genotype that was closely aligned with others. Gene sequencing similarity revealed 99% identity with other isolates deposited in GenBank. Conclusion: The occurrence of toxA was most prevalent. One isolate was recorded as a novel isolate in the global gene bank as a locally isolated strain from the city of Erbil that has never been identified in global isolates due to genetic variation.

High efficiency of Ag0 decorated Cu2MoO4 nanoparticles for heterogeneous photocatalytic activation, bactericidal system, and detection of glucose from blood sample.

The novel Ag0/Cu2MoO4 nanoparticles was simply synthesized via chemical method. Ag/Cu2MoO4 nanoparticles was characterized by FESEM image, XRD curve, UV-vis spectroscopy, BET analysis, and XPS spectrum. XRD pattern depicts that the cubic crystalline phase of particles. The band gap of Ag/Cu2MoO4 nanoparticles was achieved to 2.04 eV, which that depicted the best activity under visible light irradiation. Ag/Cu2MoO4 nanoparticles exhibits 99.74% degradation under light and persulfate ion which was higher response than Cu2MoO4 nanoparticles (83.56%) under this condition. The scavenging test indicates the important reactive species in removal process were •OH, and •SO4-. The Ag/Cu2MoO4 nanoparticles was indicated highly photo-stability for the MG degradation after 5th cycle. Ag/Cu2MoO4 exhibits substantial antibacterial properties against P. aeruginosa and S. pneumoniae. Moreover, Ag/Cu2MoO4 nanoparticles was experimented to peroxidase-like performance for the colorimetric detection of glucose with the Limit of Detection about 52.23 nM.

Synergistic antibacterial activity of silver nanoparticles biosynthesized by carbapenem-resistant Gram-negative bacilli.

Nanotechnology is being investigated for its potential to improve nanomedicine for human health. The purpose of this study was to isolate carbapenemase-producing Gram-negative bacilli (CPGB), investigate the presence of carbapenemase resistance genes, determine their antibiogram and ability to biosynthesise silver nanoparticles (Ag NPs), and estimate the antibacterial activity of Acinetobacter baumannii-biosynthesised Ag NPs on CPGB alone and in combination with antibiotics. A total of 51 CPGBs were isolated from various specimens in the study. The automated Vitek-2 system was used to identify and test these strains' antimicrobial susceptibilities. The carbapenemase resistance genes were identified using a polymerase chain reaction (PCR). Under the CPGB, A. baumannii could biosynthesise Ag NPs. X-ray diffraction (XRD), Fourier-transform infrared spectroscopy (FT-IR), transmission electron microscopy (TEM), and field emission scanning electron were used to characterise Ag NPs. The antibacterial activity of Ag NP alone and in combination with antibiotics against CPGB was determined using the broth microdilution method, and their synergistic effect was determined using the checkerboard assay. bla NDM and bla OXA-48 were the most commonly reported, and 90% of the isolates produced multiple carbapenemase genes. Tigecycline proved to be the most effective anti-CPGB antibiotic. Isolates with more resistance genes were more resistant to antibiotics, and isolates with three genes (42%) had the most extensively drug-resistant patterns (38%). A significant relationship was discovered between genetic and antibiotic resistance patterns. Only A. baumannii produced Ag NPs out of all the isolates tested. Ag NPs with a size of 10 nm were confirmed by UV-visible spectroscopy, FT-IR, XRD, and TEM analysis. The Ag NPs were effective against CPGB, with minimum inhibitory concentrations ranging from 64 to 8 µg/ml on average. Surprisingly, the combination of Ag NPs and antibiotics demonstrated synergistic and partial synergistic activity (fractional inhibitory concentration between 0.13 and 0.56) against CPGB, as well as a significant reduction in antibiotic concentrations, particularly in the case of A. baumanii versus ceftriaxone (1024 to 4 µg/ml). The notable synergistic activity of Ag NPs with antibiotics represents a valuable nanomedicine that may find clinical application in the future as a combined remedy.

Investigation on prevalence, risk factors, and genetic diversity of Pediculus humanus capitis among primary school children.

Pediculosis is an integumentary disease caused by the ecto-parasite Pediculus humanus capitis, which infests human hair. It is a common public health problem that is most prominent worldwide in elementary school children. The current study aimed to investigate the prevalence, risk factors, and genetic diversity of P. humanus capitis among primary school children in the Erbil province. For this purpose, this study was conducted from October 2019 to December 2019 among 1100 randomly selected elementary school children aged 6-12. Data collection was performed via a regular questionnaire and physical hair examination. For the genetic diversity part, after collecting one louse randomly from each individual, DNA was extracted. The mitochondrial Cox1 gene was then amplified by universal primer and PCR. Gene sequencing was performed by ABI (BioNEER, South Korea). Data analysis was done by Chi-Square and T-test using the SPSS ver. 23. The overall infestation rate was 21.27%, and the rate was significantly higher among females (34.93%) compared to males (7.91%). Some variables had found the prevalence rate to be strongly affected. This included age; the rate was not significant (26.87%) in the age group 8-9 years compared to other age groups. According to hair length, the rate was significantly increased (36.52%) among children with tall hair. In terms of hair type, the incidence of curly-haired children was significantly higher (31.54%); in terms of hair color, there were not significant differences among blonde children (25.90%) and others. According to the results of Cox1 gene sequencing, of 234 infested children to lice, 86 (36.75%) of them were exposed to clade A, 38 (16.24%) were exposed to clade B, clade C has not been seen among any children (0%), 105 students (44.87%) were exposed to clade D, and 5 of them exposed to clade E (2.14%). Eventually, a significantly higher incidence (33.78%) was reported in rural primary school children. The infection rate of human head lice in Erbil province is still high, which is one of the health problems of children in public schools.

Diagnosis and detection of VicK gene in Streptococcus mutans isolated from the saliva of patients with diabetic type 2 with tooth decay in the Iraqi population.

Type 2 diabetes mellitus (T2DM) is gradually becoming more common in Iraq. Salivary changes and proliferation of specific bacterial communities cause oral disease that can adversely affect systemic conditions such as diabetes. Fifty saliva samples were collected from people with T2DM suffering from tooth decay and twenty-five people without T2DM suffering from tooth decay. The periodontal status, the extent of the root surface, and coronal caries were evaluated. Saliva was cultured for investigating Streptococcus mutans. The results showed that patients with type 2 diabetes had significantly more severe Periodontitis and a higher prevalence and magnitude of bacterial caries. Diabetic subjects had higher levels of Hemoglobin A1c (HbA1c) and Random Blood Sugar (R.B.S.). The S. mutans diagnosis by PCR for Sanger Sequencing technique by using VicK gene sequences (1300bp). The PCR products of the isolate were submitted to Macrogen Company for sequencing. Selected seven isolates as new isolates registered in global gene bank as locally S. Mutans isolates in Bagdad city/Iraq and their accepted accession numbers include LOCUS MT603520, MT603521, MT603522, MT603523, MT603524, MT603525,and MT603526 of nucleotide sequence. The VicK genes isolates' phylogenetic trees revealed a genotype that was closely connected to other isolates in GenBank. Furthermore, gene sequencing demonstrated a success rate of 99 percent. resemblance to other isolates in the GenBank database The likelihood of a link between S. Mutans and dental carries was determined by these findings.

Evaluation of blaGES-5 and bla veb-1 genes with multidrug-resistant extend, pandrug resistance patterns (MDR, XDR, PDR), and biofilm formation in Pseudomonas aeruginosa isolates.

Pseudomonas aeruginosa is a ubiquitous microorganism that is difficult to treat due to the increasing prevalence of multidrug resistance patterns. A total of 227 samples were taken from different clinical samples during the study period from January 2018 to December 2018. The isolates were identified with antibiotic sensitivity testing with ESBL by the Vitek-2 automated method. MDR, XDR, and PDR were determined. 40 (17.6%) isolates were positive for P. aeruginosa, maximum of ESBL and MBL. Positive isolates were detected in the burn, coexisting ESBL + MBL enzymes in 21 (52.5%) of our isolates. Imipenem followed by Meropenem were found to be effective against ESBL and MBL producers. Resistance was reached between 72-100% to 5 antibiotics. The frequency of PDR, MDR, and XDR were 5%, 50%, and 45%, respectively. The frequency of co-production between MDR, XDR, and PDR with MBL, ESBL, and Biofilm was 35%, 12.5% and 5%, respectively. Among the ESBLs, the frequency of distribution of bla VEB-1gene and blaGES-5 gene was 50% and 40 %, respectively. Bacterial isolates simultaneously carrying blaVEB-1 gene with multiple ß-lactamases of different classes of biofilm, MDR, PDR, and XDR as same as a coexisting blaGES-5 gene. One isolate was detected as new isolates registered in global gene bank as locally P. aeruginosa isolates in Erbil city (LOCUS MN900953). The phylogenetic trees of the blaVEB gene isolates were demonstrated a genotype closely related to others, deposited in GenBank similar to the P. aeruginosa gene; gene sequencing revealed a 99% similarity with other isolates deposited in GenBank.

ZnO, TiO2 and Ag nanoparticles impact against some species of pathogenic bacteria and yeast.

The economic approaches for manufacturing the nanoparticles with physical and chemical effects and limited resistance to antibiotics have been progressed recently due to the rise of microbial resistance to antibiotics. This research aimed to study the antimicrobial efficacy of silver nanoparticles Ag, ZnO, and Tio2 nanoparticles against Salmonella typhimurium and Brucella abortus and Candida albicans. Two isolates of Salmonella and two isolates of Brucella abortus were isolated from food spastically meat and blood specimens, respectively. Candida albicans were isolated from the patient's mouth with oral candidiasis (oral thrush) and confirmed diagnosis by API 20C test. The antimicrobial susceptibility of Salmonella typhimurium and B. abortus isolates were performed against nine different antibiotics. Silver nanoparticles consisting of AgNPs size (90) nm, ZnO NPs size (20, 50) nm as well as TiO2 NPs size (10, 50) nm, were used. UV-Visible spectrophotometer was used to characterize silver nanoparticles. The highest resistance of Candida albicans was seen for fluconazole, Clotrimazole and Itraconazole. The results of the Minimum Inhibitory Concentration (MIC) of nanoparticles against Salmonella typhimurium showed the average MIC of Tio2-10nm and Tio2-50nm were 5000 and 2500 µg\ml for S1 and S2 isolates, respectively. The isolated Brucella abortus (B1 and B2) showed sensitivity to NPs with different MIC. The average MIC for Ag-90nm was 5000 and 2500 µg/ml for B1 and B2 isolates, respectively. The findings suggest NP solution has fungicidal and bactericidal impacts on the tested microorganisms so they can be suitable for multiple applications of the biomedical field such as developing new antimicrobial agents.

Prevalence and characterisation of carbapenemase encoding genes in multidrug-resistant Gram-negative bacilli.

BACKGROUND: Emerging worldwide in the past decade, there has been a significant increase in multidrug-resistant bacteria from serious nosocomial infections, especially carbapenemase-producing Gram-negative bacilli that have emerged worldwide. The objective of this study is to investigate carbapenem resistance in Gram-negative bacilli bacteria using phenotypic detection, antimicrobial resistance profiles and genotypic characterisation methods. METHODS: 200 Gram-negative bacilli isolates were collected from different clinical specimens. All clinical samples were exposed to isolation and identification of significant pathogens applying bacteriological examination and an automated Vitek-2 system. The isolates were subjected to susceptibility tests by the Vitek-2 automated system and those isolates that were resistant to beta-lactam drugs, including carbapenems, third-generation cephalosporines or cefoxitin, were selected for phenotyping using Carba plus disc system assay for detection of carbapenemase-producing isolates. These isolates were further confirmed by molecular detection. PCR was used for the detection carbapenem-resistant genes (OXA-48, IMP, NDM, VIM, and KPC). RESULTS: 110 (55%) of 200 Gram-negative bacilli were identified as beta-lactam-resistant isolates. The frequency of carbapenem-resistant isolates was calculated to be 30.9% (n = 34/110). A collection totalling 65/110 (59%) isolates were identified as carbapenemase producers by phenotypic method. Moreover, among the 65 carbapenemase-producing Gram-negative isolates with a positive phenotype-based result, 30 (46%), 20 (30%) and 18 (27%) isolates were positive for OXA-48, KPC and MBL enzymes, respectively, as well as the production of 27% of AmpC with porin loss. Tigecycline was the most effective antibiotic that affected 70% of MDR isolates, but high rates of resistance were detected to other tested antimicrobials. Of interest, a high incidence of MDR, XDR and PDR profiles were observed among all carbapenemase-producing isolates. 36% (24/65) of the tested isolates were MDR to 3 to 5 antimicrobial classes. 29% (17/65) of the recovered isolates were XDR to 6 to 7 antimicrobial classes. Alarmingly, 24% (16/65) of isolates displayed PDR to all the tested 8 antimicrobial classes. Genotype assay, including 53 phenotypically confirmed carbapenemase-producing isolates of Gram-negative bacilli, found 51(96%) isolates were harbouring one or more genes. The most common carbapenemase gene was bla NDM 83% (44/53) followed by bla OXA-48 75% (40/53), bla VIM 49% (26/53) and bla IMP 43% (23/53), while the gene bla KPC was least frequent 7% (4/53). 92% (46/51) of isolates were involved in the production of more than one carbapenemase gene. CONCLUSION: This study demonstrated the emergence of carbapenemase-producing Gram-negative pathogens implicated in healthcare-related infections. Accurate identification of carbapenem-resistant bacterial pathogens is essential for patient treatment, as well as the development of appropriate contamination control measures to limit the rapid spread of pathogens. Tigecycline exhibited potent antimicrobial activity against MDR, XDR and PDR-producing strains that establish a threatening alert which indicates the complex therapy of infections caused by these pathogens.

Biosynthesis and characterization with antimicrobial activity of TiO2 nanoparticles using probiotic Bifidobacterium bifidum.

Bifidobacterium selectively colonizes the infants' intestinal tract, and the relevant coliform bacteria in adults are particularly beneficial because of their enhanced capability to prevent pathogens of gastro intestine by direct antimicrobial action and relieve infection, which led to their intensification, the antibacterial activities of titanium nanoparticles producing by some bacteria, makes them attractive as a new agent against pathogenic bacteria. In our present study, we used a probiotic bacteria Bifidobacterium bifidum which was isolated from the commercial market capsule to produce TiO2 nanoparticles and study the biologically characterized nanoparticle using various techniques like Scanning Electron Microscopic (SEM), atomic force microscopy (AFM), and study its antimicrobial activity against a bacteria isolated from the stool of patients suffering from acute diarrhea. The results showed that the morphological characteristics of nanoparticles were found to have a spherical shape and mean size of 81 nm by AFM while scanning electron microscope viewed as an oval shape with anatase form synthesized by B. bifidum. TiO2-NP synthesized by B. bifidum had an inhibitory effect against P. aeruginosa, A. baumanii, K. pneumonia at a concentration 16 mg/ml and 32 mg/ml towards E. coli and S. typhi, the minimum inhibitory concentration (MIC) against pathogenic bacteria isolated from acute diarrhea included Pseudomonas aeruginosa, Acinetobacter baumanii, Klebsiella pneumonia, E.coli and salmonella typhi was utilized to determine the antibacterial impact of the synthesized TIO2 nanoparticles. Our biologically synthesized titanium nanoparticles were effective against all the tested pathogenic bacteria at various degrees and had a probable role in significantly greater antimicrobial efficacy against all isolates under study. This trial may have considerable significance for the prevention of antibiotic resistance associated diarrhea in hospitals.