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.
One of the most important virulence factors in Pseudomonas aeruginosa is biofilm formation, as it works as a barrier for entering antibiotics into the bacterial cell. Different environmental and nutritional conditions were used to optimize biofilm formation using microtitre plate assay by P. aeruginosa. The low nutrient level of the medium represented by tryptic soy broth (TSB) was better in biofilm formation than the high nutrient level of the medium with Luria Broth (LB). The optimized condition for biofilm production at room temperature (25 °C) is better than at host temperature (37 °C). Moreover, the staining with 0.1% crystal violet and reading the biofilm with wavelength 360 are considered essential factors in
... Show MorePeroxidase is a class of oxidation-reduction reaction enzyme that is useful for accelerating many oxidative reactions that protect cells from the harmful effects of free radicals. Peroxidase is found in many common sources like plants, animals and microbes and have extensive uses in numerous industries such as industrial, medical and food processing. In this study, P. aeruginosa was harvested to utilize and study its peroxidases. P. aeruginosa was isolated from a burn patient, and the isolate was verified as P. aeruginosa using staining techniques, biochemical assay, morphological, and a sensitivity test. The gram stain and biochemical test result show rod pink gram-ne
... Show MoreIn this study, 158 clinical samples were collected from hospitalized burn patients during the period from December 2012 to June 2013 in Karbala province\ Iraq. Bacterial isolates were identified using conventional biochemical tests and then identification was confirmed by using Vitek-2 compact system. Pseudomonas aeruginosa recovery was 60 isolates in this study. These isolates were analyzed for antibiotic susceptibility by the disk diffusion test (DDT) according to Kirby Bauer's method using seven clinically important antipseudomonal agents: carbapenems (Imipenem and Meropenem), pencillins (Piperacillin), cephalosporins (Ceftazidim), monobactam (Aztreonam), quinolones (Ciprofloxacin) and aminoglycosides (Gentamicin). The results of resista
... Show MoreBackground: L. sativum, are traditionally used for the treatment of various diseases and thought to have medicinal value. Isolates from many part of the world is now multidrug resistant. Therefore, there is an urgent need to look for and test an alternative herbal drug.
Objective: The present study aimed to evaluate the antibacterial activity of L. Sativum seed extract against multi drug resistant (MDR) and sensitive Pseudomonas aeruginosa clinical isolates.
Subjects and Methods: An ethanolic and aqueous stock extracts were prepared from L. sativum seed plant then serial dilutions were prepared and the obtained concentrations (50, 25, 12.5 and 6.2 mg/ml) were tested against 30 multidrug-resistan
... Show MoreIntroduction: Melanin is a high-molecular weight pigment produced through the oxidative polymerization of phenolic or indolic compounds and plays a perfect role in UV-light shielding, as well as in photoprotection. Among biopolymers, melanin is unique in many aspects. This study is designed to screen Production, extraction and characterizes of an extracellular melanin pigment from clinically isolated P. aeruginosa. Objective: The aim of the current study is isolation and diagnosis of P.aeruginosa using vitek-2 compact system and screening the ability to produce melanin and characterization of extracted melanin by UV-vis, FTIR, XRD and SEM. Materials and methods: the samples swab inoculated on cetrimide agar as selective media and incubated
... Show MoreBackground: Acute urinary tract infection is a common bacterial infection causing illness in infants and children. At age of seven, 8% of girls and 2% of boys will have at least one episode. Although drinking water and using home remedies are known to help to flush away bacteria and keeps them from sticking to the bladder wall, researches to test the efficacy and safety of hydrochlorothiazide's diuretic effect as adjuvant to the antibiotics in pediatric age groups are lacking, and so this research was to address this subject.
Objectives: To assess the effectiveness and the safety of hydrochlorothiazide as adjuvant therapy to the antibiotics in treating acute urinary tract infect
... Show MoreKlebsiella pneumoniae causes lethal nosocomial infections, mostly affecting patients with severe burns. More than 80% of its isolates have shown resistance to routinely used antibiotics in parallel with increased infection rates. The study aimed to determine the molecular typing and genetic relatedness of K. pneumoniae. Therefore, 20 multidrug resistant (MDR) K. pneumoniae already isolated from infected burned wounds in two major hospitals of Al-Kut city east Iraq were subjected to genotyping analysis. The random amplified polymorphic DNA (RAPD)-based polymerase chain reaction (PCR) technique was used along with three oligonucleotide primers (P13, OPX-04, and OPY-01). The amplicons’ patterns of the electrophoresis-gel were analyzed by the
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