Respiratory tract infections in sheep are among the important health problems that affect all sheep ages around the world. Nine bacterial isolates obtained from sheep with respiratory tract infections were selected to be used in the current study. The isolates included 3 Staphylococcus aureus, 4 Klebsiella pneumoniae, and 2 Pseudomonas aeruginosa. Following the primers design by the Primer3Plus software tool and optimization of the conventional polymerase chain reaction (PCR), the primers were validated for their use in the multiplex PCR experiments. The MFEprimer program was used to check the suitability of the primer set combinations for multiplex PCR. The MFEprimer software was successful in designing the multiplex-PCR experiments and determining the optimal primer set combinations. Multiplex PCR was able to amplify specific DNA sequences of one, two or three target genes of these mixed microorganisms in the same PCR reaction tube. This technique efficiently detected combinations of two organisms, either S. aureus with K. pneumoniae, S. aureus with P. aeruginosa or K. pneumoniae with P. aeruginosa. Moreover, multiplex PCR was also able to detect the presence of the three organisms together in the same reaction tube. To conclude, this study confirmed multiplex-PCR as a specific, sensi- tive, rapid, accurate, and cost-effective molecular diagnostic method for identification and differentiation of three clinically important bacteria associated with sheep respiratory tract infections, including S. aureus, P. aeruginosa, and K. pneumoniae. This can efficiently support control and treatment of such diseases and would increase the economy of the animals’ owners and wellbeing of the animals.
The increasing use of antiseptic compounds creates selective pressure cause emergence of antiseptic resistance among Staphylococcus aureus .Resistance mechanism of antiseptic is driven mainly by multi drug resistant (MDR) efflux protein.Sixty five isolates of S.aureuswere collected from different clinical sources and subjected to 11 antibiotics most of them are recognized by efflux systems as extruded substrates. Range of efflux activity was estimated using cartwheel method. Simultaneous discrimination of antiseptic coding genes (qacA/B, smr and norA)as well as nuc and mecA genes among multidrug resistantS.aureus(MRSA) isolates was preformed using multiplex PCR assay
... Show MoreNowadays, there is increased interest in the biosynthesis of microbial melanin related to their numerous biological functions and applications in many fields, especially in medical fields, including immune-modulating, antimicrobial antibiotic, antiviral antivenin, anticancer, antitumor activity, and anti-biofilm activity. Pyomelanin is a hydrophobic macromolecule that is typically dark brown or black in color, formed by the oxidative polymerization of phenolic or indolic compounds. Pyomelanin is reported to be safe for consumption, thus providing a crucial strategy for biocontrol of biofilm. Furthermore, natural pyomelanin is known as a potent antioxidant, photoprotective, and free radical scavenging. Objective: This study focuses on the
... Show MoreThe isolates of Staphylococcus aureus were isolated from patients with various infections in hospitals, the isolates were identified and accurately diagnosed by phenotypic examination and biochemical tests, as well Vitek-2, and then genetic detection and diagnosis of many of the pathogenic factors associated with Staphylococcus aureus using conventional polymerase chain reaction (PCR) and testing for association by antibiotic resistance and production of some toxins by Staphylococcus aureus. After performing analysis of statistical, it was set up that the correlation coefficient of the PCR technique using virulence genes, sensitivity test to antibiotics and other virulence factors were significant at p < 0.05, but was insignificant with the
... Show MoreThis study investigated the prevalence of quinolones resistance proteins encoding genes (qnr genes) and co-resistance for fluoroquinolones and β-lactams among clinical isolates of Klebsiella pneumoniae. Out of 150 clinical samples, 50 isolates of K. pneumoniae were identified according to morphological and biochemical properties. These isolates were collected from different clinical samples, including 15 (30%) urine, 12 (24%) blood, 9 (18%) sputum, 9 (18%) wound, and 5 (10%) burn. The minimum inhibitory concentrations (MICs) assay revealed that 15 (30%) of isolates were resistant to ciprofloxacin (≥4µg/ml), 11 (22%) of isolates were resistant to levofloxacin (≥8 µg/ml), 21 (42%) of isolates were re
... Show MoreOne hundred and eighty five urine samples were collected eight isolates (4.3%) were obtained and diagnosed as Staphylococcus aureus. Among 8 isolates, 5 (62.5%) S. aureus isolates were found to be enterotoxigenic, most of isolates produced at least two types of Staphylococcal enterotoxins (SEs). The production of enterotoxins in the presence or absence of Thymol extracts (aqueous and alcoholic) were estimated using a reversed passive latex agglutination (SET-RPLA) kit. The extracts reduced enterotoxin production compared with the control. Enterotoxin inhibition was observed for enterotoxin C production at minimal inhibitory concentrations (MIC) at 400 µg/ml, whereas production of enterotoxins A, B, and
... Show MoreForty five wound specimens were collected from patients suffering from wound infections and taken from various hospitals in Ibb city, Yemen. The study was to determine synergic antibacterial activity of between mountain honey and Argemone mexicana plant. Isolation, identification of bacterial isolates and antibiotic sensitivity test were done. Agar-disc and agar-well diffusion method were carried to determine antibacterial activity of honey, Argemone mexicana plant and a mixture of them against bacterial isolates. Out of 45 specimens, 29 (64.4%) gave positive cultures. Staphylococcus aureus was the predominant bacterial pathogens with percentage (72.4%) followed by Pseudomonas aeruginosa (17.2%) and Staphylococcus epidermidis (10.4%).
... Show MorePetroleum is one of the most important substances consumed by man at present times, a major energy source in this century, petroleum oils can cause environmental pollution during various stages of production, transportation, refining and use, petroleum hydrocarbons pollutions ranging from soil, ground water to marine environment, become an inevitable problem in the modern life, current study focused on bioremediation process of hydrocarbons contaminants that remaining in the bottom of gas cylinders and discharged to the soil. Twenty-four bacterial isolates were isolated from contaminated soils all of them gram negative bacteria, bacterial isolates screening to investigate the ability of biodegradation of hydrocarbons, these isolates inocula
... Show MoreIntroduction and Aim: Pseudomonas aeruginosa is a nosocomial infection with an ability to develop high levels of antibiotic resistance. The efflux pump system is one of the mechanisms that is linked to multidrug resistance in P. aeruginosa. In this study, we employed siRNA loaded on gold nanoparticles against the MexA efflux pump gene to decrease the MexA gene expression in P. aeruginosa and estimated antibiotic resistance after gene silencing. Materials and Methods: This study examined four strains of P. aeruginosa isolated from patients in various hospitals in Baghdad. Bacteria isolated were identified by biochemical tests and Vitek compact 2 system. Single-stranded siRNA (33bp) designed in this study was loaded onto gold
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