Identification of Burkholderia mallei Isolates with Polymerase Chain Reaction-Restriction Fragment Length Polymorphism.

Burkholderia mallei is the main cause of glanders as a dangerous contagious zoonosis disease that is mostly observed in single-hoofed animals, especially horses. Modern molecular techniques have been recently employed to improve epidemiology for identifying and searching for strains of this bacterium at different times and locations. Due to the unknown number of circulating strains and lack of preventive methods, glanders is still observed in the form of epidemics. The present study aimed to evaluate six field isolates plus two laboratory strains of Borkolderia mallei and Burkholderia pseudomallei using the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method. All the isolates and strains were microbially cultured in the glycerol nutrient and glycerol agar media. The individually grown colonies of the bacterium were used in the biochemical tests. The DNA of isolates was extracted by boiling, and the PCR-RFLP test was conducted on their genome. Finally, the bacterium was injected into guinea pigs to induce the Straus reaction. The biochemical assays (or bioassays) confirmed the isolates as Burkholderia mallei. The PCR-RFLP assay demonstrated a product for Burkholderia mallei with a length of 650 bp. Nevertheless, 250 and 400 bp were produced for Burkholderia pseudomallei. The swollen scrotum pointed to the occurrence of the Straus reaction. The PCR-RFLP is a proper differential diagnosis technique for B. mallei; moreover, it is a suitable method for differentiating between Burkholderia mallei and Burkholderia pseudomallei. This technique can detect Burkholderia mallei in a short time with high precision and sensitivity.

Combined Molecular and Biochemical Identification of Alpha Toxin in Local Isolated Clostridium novyi from the Sheep Liver.

Clostridium novyi (C. novyi) causes deadly Black disease in sheep and rarely in other animals. Alpha toxin (α-toxin), the most apparent pathogen of this disease, is produced by C. novyi type B. Economic damages of C. novyi include sheep mortality costs, depreciation of affected farms, and health problems with infected carcasses. The identification of C. novyi and isolation of its pathogens by conventional methods is a time-consuming process, necessitating a simple and rapid method for isolating and detecting pathogenic C. novyi. Therefore, this study aimed to molecularly identify α-toxin in local C. novyi isolates from the sheep livers. In this study, 75 livers suspected of Black disease were sampled. The samples of the liver were cultured under anaerobic conditions. Some of the cultured colonies were used in biochemical tests. For molecular confirmation, the DNA of isolates was extracted, and the isolates were confirmed by the polymerase chain reaction (PCR) on the liver tissue and cultured samples using specific α-toxin primers. The PCR on α-toxin produced a band in the range of 609 bp, indicating that the samples belonged to C. novyi. According to the results, of 75 isolates, 18 isolates were confirmed as C. novyi. C. novyi type B was isolated from the liver and confirmed by biochemical and molecular characterization. The PCR assay ensured a sensitive and specific tool for the detection of C. novyi in the samples.

Investigation of microstructure evolution and martensite transformation developed in austenitic stainless steel subjected to a plastic strain gradient: A combination study of Mirco-XRD, EBSD, and ECCI techniques.

The evolution of the microstructure and deformation mechanism at different levels of plastic strain are investigated for 304 L austenitic steel with a combination of micro X-ray diffraction (XRD), electron backscattered diffraction (EBSD) and electron channeling contrast imaging (ECCI). A plastic strain gradient is developed along the longitudinal rolling direction in a wedge-shaped 304 L austenitic steel sample. The graded deformed microstructure includes various amounts of deformation bands, ε- and α'-martensites, and their intersections form during the plastic deformation. ECCI observations reveal several deformation mechanisms of the formation of partial dislocations, dislocations, and interactions. This study suggests geometrically necessary deformation bands (GNDBs) are introduced and stored instead of geometrically necessary dislocations (GNDs) in low stacking fault energy (SFE) materials such as austenite steels. Consequently, increasing the strain gradient leads to an increase in the geometrically necessary martensitic transformation (GNMT); this is the result of the deformation-induced martensite in these materials. In addition to the statistically stored dislocations (SSDs), GNDs are generated at the grain boundaries of the fragmented grains to preserve the continuity of the grains. Accordingly, the strain hardening of the austenite steel includes multiple interactions of the deformation bands, SSDs, GNDs, GNDBs, and GNMT. From the viewpoint of microstructure design, our study provides quantitative information about the relationship between the amount of plastic deformation and the extent of microstructure evolution, in a continues design space.

Detection and Isolation of Mycoplasma capricolum Subspecies Capricolum from East Azerbaijan Sheep Flocks.

Mycoplasma capricolum subspecies capricolum (Mcc) is one of the causative agents of contagious agalactia (CA), which is an important disease in sheep and goats in the Mediterranean and Middle East countries. Mycoplasma agalactiae is the classic agent of CA in sheep and goats. Mycoplasma mycoides subspecies Capri (Mmc), Mycoplasma capricolum subspecies capricolum (Mcc), and Mycoplasma putrefaciens (Mp) produce a clinically similar disease, more often in goats. The aim of the present study was to detect Mcc in sheep flocks in East Azerbaijan Province of Iran. Milk, ear canal, and eye swab samples were collected from 49 sheep flocks with clinical signs of CA or a history of a disease. All the samples were examined using both culture and molecular methods. In the molecular method,positive samples for the Mycoplasma genus were tested for M. mycoides cluster and Mcc. From 272 samples, 67, 87, and 62 samples were shown to be positive using the culture method, polymerase chain reaction (PCR) method, and both culture and PCR methods, respectively. Mcc was detected in all the four M. mycoides cluster positive samples, including milk, ear canal, and eye swab samples. This is the first report of Mcc detection from East Azerbaijan. Our results showed that eye, milk, and ear canal samples could be suitable sources for Mcc detection in sheep flocks.

Molecular Identification of Extended-Spectrum ß-lactamase and Integron Genes in Klebsiella Pneumonia.

INTRODUCTION: Infections caused by Gram negative bacteria, producing extended-spectrum ß-lactamase, including Klebsiella pneumoniae are increasing all over the world with high morbidity and mortality. The aim of the present study was determined antimicrobial profile susceptibility and the prevalence of antibiotic resistance genes by multiplex PCR. METHODS: In the present study, we obtained one-hundred isolates of K. pneumoniae from different clinical samples. The antibiotic susceptibility testing was done in thirteen antibiotic and, therefore, M-PCRs were conducted using the DNA amplification for detection of ESBLs (blaTEM, blaCTX-M, blaSHV) and int (I, II, III) genes. RESULTS: The results of resistance to amoxicillin/clavulanate, ciprofloxacin, amikacin, trimethoprim-sulfamethoxazole, cefotaxime, ampicillin, aztreonam, imipenem, gentamicin, ceftazidime, Cefepime, ceftriaxone and levofloxacin were obtained 37%, 37%, 93%, 84%, 52%, 87%, 59%, 8%, 24%, 67%, 52%, 43% and 26%, respectively. The frequency of the extended-spectrum ß-lactamase K. pneumoniae was obtained 37%. The prevalence of resistance genes of ESBLs in the M-PCR method showed that the blaTEM, blaCTX and blaSHV were 38%, 24% and 19%, respectively, however, only 8 (8%) out of 100 isolates were found to have positive outcomes for the existence of class 1 integrons and there were no detected class 2 or class 3 integrons. CONCLUSIONS: Our results recommend the likely co-carriage of some ESBLs genes and antibiotic resistance integrons on the same plasmids harboring the MDR genes.

Pooling batches in drug stability study by using constant-width simultaneous confidence bands.

One important study objective in drug stability studies is to estimate the shelf-life of a drug. A key statistical problem involved in this is how to assess the practical equivalence of different batches of the same drug so that different batches can be subgrouped to produce a single shelf-life for the drug. In this paper constant-width simultaneous confidence bands are proposed to quantify the magnitude of difference between different batches, with a particular view to establish the practical equivalence of different batches. This approach is suitable for the situation that the intercepts and slopes of the regression lines for the batches cannot be assumed to be equal. It is shown how constant-width simultaneous confidence bands can be easily constructed for the multiple comparison of several general linear regression models. In particular, it is shown that constant-width simultaneous confidence bands have a better chance to establish the equivalence than, and so are preferable to, the hyperbola-shaped simultaneous confidence bands considered.

The role of outer membrane proteins of Ornithobacterium rhinotracheale in attachment to chicken tracheal epithelium.

Ornithobacterium rhinotracheale (ORT) infections cause major losses to the poultry industry. In search for factors implicated in the pathogenesis of ORT infections, the role of outer membrane proteins (OMPs) in the interaction of ORT with chicken tracheal epithelium was investigated. For this purpose, immune sera were prepared against total extracted OMPs, whole cell bacteria and three major OMPs of 45, 53 and 70 kDa and used in bacterial adherence inhibition assay. The results showed antibodies against a 53 kDa OMP significantly (p < or = 0.05) inhibited the bacterial adherence to chicken tracheal epithelium up to 78%.

Beneficiary effect of dietary soy protein on lowering plasma levels of lipid and improving kidney function in type II diabetes with nephropathy.

OBJECTIVE: Heart and renal diseases are two major problems in diabetic patients. Hyperlipidemia is one of the main risk factors of cardiovascular complications in diabetes. The type of protein consumed also affects the changes in renal blood flow, glomerular resistance and renal function in these patients. Hence, this study was undertaken to show the effect of soy protein consumption on lipid profiles and kidney function of diabetic patients with nephropathy, who attended an educational university hospital as well as a private kidney disease clinic in Tehran. SUBJECTS AND METHODS: This crossover randomized clinical trial was conducted on 14 patients who were free of any uncontrolled condition or other renal diseases. The patients were asked to follow a usual nephropathy diet (0.8 g/kg protein, 70% animal and 30% vegetable protein) for 7 weeks. After a washout period of 4 weeks consuming the prestudy diet, subjects were readmitted to repeat the same cycle with a similar diet containing 35% soy protein and 30% vegetable protein. Paired t-test, carryover effect and period effect were used for statistical analysis. RESULT: : There were 10 men and four women whose mean (s.d.) of weight was 70.6 (10.3) kg. Significant reductions were seen in total cholesterol (P<0.01), triglyceride (P<0.002) and LDL-c (P<0.04), urinary urea nitrogen and proteinuria (P<0.001) after soy vs animal protein consumption. There were no significant changes in HDL-c, LDL-c/HDL-c levels. We also saw a favorable effect on renal function. CONCLUSION: Soy inclusion in the diet can modify the risk factors of heart disease and improve kidney function in these patients.