Salmonella is widely distributed throughout the world and can be found in poultry industry, animal breeding centers, food and feedstuffs of all geographical regions. This study was conducted to determine and identify Salmonella serovars isolated from poultry, calves and foodstuffs (poultry and animals products such as egg and meat). A total of one hundred isolates of Salmonella serovars including Salmonella Typhimurium, Salmonella Enteritidis, Salmonella Infantis, Salmonella Gallinarum and Salmonella Pullorum consecutively were subjected to the conventional culture, biochemical and serological assays. The utility of molecular multiplex PCR was investigated to identify and differentiate among five Salmonella serovars which were identified according to the presence of rfbJ, fljB, invA, and fliC genes in S. Typhimurium, sefA, invA and spv genes in Salmonella Enteritidis, fljB, fliC and invA genes in Salmonella Infantis, hut and slgC genes in both Salmonella Gallinarum and Salmonella Pullorum and speC gene specifically in Salmonella Gallinarum. Biochemical assays and serotyping are complicated to directly differentiate between Salmonella Gallinarum and Salmonella Pullorum because of their antigenic similarity. According to the results, Multiplex PCR can be considered as simple, rapid, accurate and useful test to identify and differentiate among Salmonella serovars.
Colibacillosis is an important disease in the poultry industry which causes serious economic damages. As it is suggested that vaccination is one of the means to control colibacillosis, we tried to investigate the vaccine potential of a ΔaroA derivative of an O78:K80 avian pathogenic Escherichia coli containing increased serum survival gene. 490 chicks were selected as follows: For assessment of virulence of ΔaroA mutant, 30 chicks were divided into three groups and injected with 0.5ml of PBS or bacterial suspension containing either10(7)colony forming units (CFU) of mutant or parent strains via subcutaneous route. Macroscopic lesions and mortality rate were recorded in different groups during the week after challenge. For assessment of safety and immunogenicity of the ΔaroA mutant, three groups of 20 chicks were vaccinated by aerosol administration of 250 ml of suspension containing 10(8) CFU of mutant strain at days 1 and 14, while the two other groups received PBS or wild type strain. Macroscopic lesions and mortality rate were recorded in different groups until day 21. To determine whether the vaccination is protective against challenges or not, the chickens were vaccinated at days 1 and 14 and challenged intramuscularly with either a homologous or heterologous strains at day 21. Macroscopic lesions and mortality rate were recorded in different groups during the week after challenge. The results revealed that the ΔaroA mutant was slightly virulent, however it was safe and did not cause mortality, lesions or weight loss after vaccination. Antibody responses were similar in the control and mutant groups and vaccination did not induce a significant humoral immunity. The mutant could not protect chickens against both homologous and heterologous challenges. This could be due to several factors such as the high amount of maternal antibodies in the first two weeks of life, and the vaccination procedure
Significance and Impact of the Study: We investigated the antibacterial, anti-biofilm and quorum sensing inhibitory properties of Thymus daenensis and Satureja hortensis essential oils against food-borne pathogen Escherichia coli O157:H7 using phenotypic and genotypic methods. Accordingly, the present study introduces T. daenensis and S. hortensis essential oils as new antimicrobial agents against E. coli O157:H7.
The nephropathogenic infectious bronchitis virus (IBV) strain IS-1494 like (variant-2; GI-23) was first isolated in the Middle East (1998). Despite intensive vaccinations, IS-1494 like IBVs are still circulating in Iran (the dominant genotype) and spread to other countries. Here, the full-length genome of this Iranian IS-1494 like IBV was (Mahed) determined to understand its evolutionary relationships. The genome consists of 27,652 nucleotides, with mutations in most of the structural genes. Thirteen open reading frames (ORFs) were predicted in the Mahed isolate (5' UTR-1a-1b-S-3a-3b-E-M-4b-4c-5a-5b-N-6b-3' UTR). ORFs 4b, 4c, and 6b, which has rarely been reported, were present in the Mahed genome. According to phylogenetic analysis of the full-length genome, 1a, S2, M, E, N protein, Mahed isolate clustered with the QX type strain. Based on the partial 1b, S1, Mahed clustered with the Q1 strain. The full-length genome of Mahed isolate shared the highest sequence homology with Gray and JMK (90.06-90.07%) and was least related to the Vic-s (86.21%). These data show that evolutionary variation because of recombination in IBV plays a major role in the adaptation and origin of IBV leading to new genetic and types of the virus strain.
Streptococcus pneumoniae (pneumococcus) is recognized as one of the major cause of infections in communities and hospitals. In this study, anti-pneumococcal and anti-efflux pump activity of two medicinal plants (Thymus daenensis and Origanum vulgare) essential oils were evaluated. Checkerboard assay test was performed for investigation of the effects of selected EOs on ciprofloxacin (CIP) and ethidium bromide (EtBr) uptake in pmrA-overexpressed fluoroquinolone-resistant pneumococcus. Using quantitative real-time RT-PCR the PmrA efflux pump gene (pmrA) expression was evaluated following treatment with selected EOs. Gas chromatography-mass spectrometry analysis was performed for identifying the major components of the tested EOs. The minimum inhibitory concentration (MIC) values for pneumococcus isolates were 0·625-2·5 μl ml for T. daenensis and 1·25-5 μl ml for O. vulgare EOs. We confirmed that in all strains T. daenensis and O. vulgare have a total or partial synergistic effects with CIP and EtBr (FICI from 0·14 to 0·75). In other hand MIC/2 concentration of T. daenensis and O. vulgare EOs caused a significant downregulation of pmrA gene (P < 0·05) in seven of eight strains. This study showed that T. daenensis and O. vulgare EOs have strong antimicrobial and anti-efflux pump activity against clinical isolates of S. pneumoniae and might be useful in controlling pneumococcal infections. SIGNIFICANCE AND IMPACT OF THE STUDY: This study introduced Thymus daenensis and Origanum vulgare essential oil as new antibacterial and anti-efflux pump agents against fluoroquinolone-resistant Streptococcus pneumoniae clinical isolates. These findings indicate that combination of these two essential oils with fluoroquinolone antibiotics may provide alternative methods to overcome the fluoroquinolone-resistant S. pneumoniae.
Objective
The objectives of this study were to investigate the antifungal susceptibility and genetic diversity of Candida albicans isolated from HIV+ patients with oropharyngeal candidiasis. A total of 50 C. albicans isolates were cultured on Sabouraud glucose agar containing chloramophenicol. The antifungal susceptibility of the isolates against fluconazole, clotrimazole, nystatin, amphotericin B, ketoconazole and flucytosine was assessed using disc diffusion method. The genetic diversity of C. albicans isolates was determined using random amplified polymorphic DNA marker.
Results
The inhibition zones ranged from 4 ± 1.8 to 40 ± 3.8 mm for fluconazole, 7 ± 1.0 to 37 ± 1.8 mm for ketoconazole, 14 ± 0.8 to24 ± 0.8 mm for amphotericin B, 25 ± 0.0 to 33 ± 0.0 mm for nystatin and 7 ± 4.2 to 40 ± 0.0 mm for clotrimazole. At 90% similarity, three distinct groups were observed. The smallest cluster composed of 3 isolates, whereas the largest one composed of 17 isolates. 32% (16/50), 28% (14/50) and 14% (7/50) were resistant to fluconazole, ketoconazole and clotrimazole, respectively.
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