BACKGROUND: Heparin is a sulfated glycosaminoglycan. Blood is a common source for DNA detection in all kinds of samples, and anticoagulants such as heparin and ethylenediaminetetraacetic acid (EDTA) are used to prevent coagulation. Because heparin has a strong inhibitory effect on polymerase chain reaction (PCR), it is not used in samples that will be tracked by DNA. There are physical, chemical, and enzymatic methods to eliminate the inhibitory effect of heparin on PCR test.OBJECTIVES: First, to compare the intensity of the inhibitory effect of two anticoagulants, heparin, and EDTA, on the Real-Time PCR (qPCR), and then to investigate the impact of the heparinase enzyme present in the medium culture extract of Pseudomonas aeruginosa, on removing the inhibitory effect of heparin during the real-time PCR.METHODS: In the present study, two blood samples containing heparin and EDTA were subjected to a real-time PCR test to check the intensity of the inhibitory effect. Then, the medium culture extract of Pseudomonas aeruginosa was added to the heparinized blood sample infected with Escherichia coli bacteria in two groups with different conditions. In the first group, the DNA in the heparinized blood sample was extracted by the phenol-chloroform isoamyl alcohol method. Then, these samples were incubated with the extract of Pseudomonas aeruginosa bacteria culture medium at different hours, but in the second group, the samples were incubated at different hours before DNA extraction. Also, the DNA concentration in both groups was measured by a Nanodrop device, and finally, all samples were subjected to a real-time PCR test.RESULTS: The results of the research samples showed that although the heparinized blood sample contains more DNA concentration than the EDTA blood sample, it completely prevents genome replication. Also, incubating heparinized blood with Pseudomonas aeruginosa culture medium extract before DNA extraction for more than 24 hours removes the inhibitory effect of heparin during the real-time PCR, even at a lower cycle threshold than the EDTA-containing sample.CONCLUSIONS: The Pseudomonas aeruginosa culture medium extract may enable researchers to use heparinized blood samples for genome amplification and diagnosis without using expensive and limited commercial heparinase enzyme.

BACKGROUND: Today, the fight against the bacteria causing foodborne diseases is of particular importance in the packaging of seafood. It is therefore vital to find new compounds with antibacterial properties.OBJECTIVES: In the present study, antibacterial properties of synthetic nanocomposite zinc chromite-zinc aluminate (ZnCr2O4-ZnAl2O4) on E. coli and Pseudomonas aeruginosa were studied.METHODS: After synthesis of nanocomposite, antibacterial activity of nanocomposite zinc chromite-zinc aluminate was evaluated via disk diffusion method, Minimum Inhibition Concentration (MIC), and Minimum Bactericidal Concentration (MBC) using the microdilution method.RESULTS: The results of this study revealed a higher sensitivity reaction of Pseudomonas aeruginosa (18.6±1.2 mm) compared to E. coli (12.7 ± 1.4 mm). No significant differences were observed between Gentamicin antibiotic and synthetic nanocomposite against Pseudomonas aeruginosa (P<0.05). The minimum MIC and MBC concentrations were seen in Pseudomonas aeruginosa (1.66 mg/ml) and the maximum concentration of MIC belonged to E. coli (5 mg/ml).CONCLUSIONS: In this study, the effects of nanoparticles on these gram-negative bacteria could be attributed to the small diameter of the ions, and hence the greater penetrability of these nanoparticles despite the wall's resistance. Based on the results, zinc chromite-zinc aluminate nanocomposite showed a better performance compared with gram-negative bacteria, specifically Pseudomonas aeruginosa resistant bacteria, and could be used for further studies in fisheries product packaging.

Periodontal diseases are the most frequently diagnosed problem in small animal veterinary medicine. Although their exact cause is not fully understood, bacteria play an important role in their development. Pseudomonas aeruginosa is a Gram-negative, rod-shaped, non-spore-forming bacterium. The living environment of this bacterium may be soil and water; however, it can also be found in humans and animals. Antibiotic treatment of periodontitis may be complicated by the carbapenem resistance of some P. aeruginosa strains, if these bacteria are found to be an aetiological agent. The aim of the study was to identify all bacterial strains isolated from dog with periodontitis.

The leaves of Bandotan (Ageratum conyzoides) are a plant thought to have antibacterial properties. This study aims to determine the sensitivity of Bandotan leaf extract in inhibiting the growth of the bacteria Pseudomonas aeruginosa. This study used a stock extract of Bandotan leaves from the Pharmacology Laboratory and a bacterial isolate of P. aeruginosa in the Microbiology Laboratory of the Faculty of Veterinary Medicine, Universitas Syiah Kuala, which was identified by Gram staining, indole test, Methyl Red test, and confectionery test. The research method was carried out by planting the re-identified bacterial isolates on Nutrient Broth (NB) media, incubated at 37C for 24 hours. Then the turbidity composition of the isolates was arranged to match the turbidity in 0.5 McFarland solution. Furthermore, the sensitivity test of the extract on Mueller Hinton Agar (MHA) media was carried out by levelling the bacterial isolates on the surface of the media and attaching a disc containing bandotan leaf extract with a concentration of 25%, 50%, 75% and gentamicin disk as a positive control and distilled water as a negative control. All treatments were incubated at 37C for 24 hours, and then the inhibition zone was measured using millimeters (mm) callipers. The results showed that concentrations of 25%, 50% and 75%, respectively, had an inhibition zone of 8.16 mm, 9.82 mm, and 16.08 mm, respectively. In contrast, the average inhibition zone for gentamicin was 25, 30 mm and 0 mm distilled water. Therefore, it can be concluded that the Bandotan leaf extract is sensitive to growth inhibition of P. aeruginosa bacteria.

Pseudomonas aeruginosa is an important animal pathogen and contributes to hemorrhagic pneumonia in mink. Between April 2011 and December 2016, samples of lung, liver, and spleen were collected from mink with this disease on 11 mink farms in 5 Chinese provinces. From these samples, we obtained 98 isolates of P. aeruginosa that belonged to 5 serotypes: G (n = 58), I (n = 15), C (n = 8), M (n = 5), and B (n = 2); 10 isolates were not typeable (10/98). More than 90% of the isolates formed biofilms, and 85% produced slime. All 98 isolates were resistant to 10 antibiotics (oxacillin, ampicillin, penicillin G, amoxicillin, ceftriaxone, cefazolin, cefaclor, tilmicosin, tildipirosin, and sulfonamide). However, almost all were susceptible to gentamicin, polymyxin B, and amikacin. We identified 56 unique genotypes by pulsed-field gel electrophoresis. These findings have revealed genetic diversity and high antimicrobial resistance in P. aeruginosa isolated from mink with hemorrhagic pneumonia and will facilitate the prevention and control of this disease.

Hemorrhagic pneumonia can be a major cause of mortality in farmed mink in the fall. In its classic form, hemorrhagic pneumonia is caused by the bacterium Pseudomonas aeruginosa. In recent years, however, outbreaks of this type of pneumonia that are associated with hemolytic Escherichia coli have also occurred in farmed mink. The purpose of this study was to compare histological lesions of acute hemorrhagic pneumonia associated with both P. aeruginosa and E. coli in mink, including a description of tissue distribution of pathogens, in an attempt to differentiate between the 2 disease entities based on histopathology. The study included material submitted for diagnostic investigation to the National Veterinary Institute in Denmark from 2006 to 2009. Altogether, 19 cases of hemorrhagic pneumonia with a pure lung culture of P. aeruginosa and 18 cases of hemorrhagic pneumonia with a pure lung culture of E. coli were examined. Formalin-fixed paraffin-embedded lung tissue obtained from the mink was examined by histology and fluorescence in-situ hybridization (FISH). It was possible to detect a slight histological difference between hemorrhagic pneumonia caused by P. aeruginosa and by E. coli, as P. aeruginosa was most often found surrounding blood vessels and lining the alveoli, while E. coli showed a more diffuse distribution in the lung tissue. Furthermore, P. aeruginosa often elicited a very hemorrhagic response in the lung, while infection with E. coli was associated with a higher frequency of alveolar edema and mild lymphoid cuffing in the lungs.

About 8 year-old castrated male Yorkshire terrier was presented for evaluation of dysuria, stranguria, hemtauria, and pollakiuria. On history taking, dysuria first was observed three months ago and these signs were waxed and waned. Physical examination revealed mild left perineal swelling. On routine laboratory examination, no significant findings were identified. Positive contrast urogram identified peritoneal herniation of urinary bladder. Urinalysis showed proteinuria and hematuria.

To analyze the major outer membrane proteins (OMPs) of the sensitive or resistant Pseudomonas aeruginosa strains, the OMPs were separated from the cellular elements by sarcosyl extraction method. OMPs profiles were conducted by SDSpolyacrylamide gel electrophoresis. Amoxicillin clavulanic acid (AMC) sensitive P. aeruginosa serotype K showed four protein bands; 35.713, 31.159, 26.107 and 22.869 KD. While AMC sensitive P. aeruginosa serotype H showed three bands of 35.713, 27.164 and 23.174 KD. Whereas AMC resistant P. aeruginosa serotype G, that was positive for the blaTEM gene by the PCR, modified its protein pattern. It has five protein bands of 52.142, 38.525, 30.690, 27.164 and 22.569 KD. These findings suggested that blaTEM gene and the outer membrane protein barrier are contributed to the resistance to amoxicillin clavulanic acid in P. aeruginosa. To determine a possible relationship between the resistance of P. aeruginosa and the production of antibodies against its outer membrane protein, antibodies against OMPs of AMC sensitive and resistant P. aeruginosa strains were prepared in mice and evaluated by ELISA. Our results showed that there was no association between immunogenicity of the outer membrane proteins and resistance of P. aeruginosa to antibiotics.

Pseudomonas aeruginosa is one of the most common pathogenic species associated with canine otitis externa (OE). Their resilience is achieved by forming a biofilm, which allows these bacteria to evade even the harshest of treatments. This study evaluated the in vitro synergistic efficacy of N-acetylcysteine (NAC) with different antimicrobial agents against P. aeruginosa isolated from dogs with OE to develop an effective treatment against P. aeruginosa. The antimicrobial activity was evaluated by the minimum inhibitory concentration test using the microdilution method. The efficacy of antibiofilm formation was evaluated using crystal violet stain method. The treatment solutions included NAC alone, and in synergy with enrofloxacin, polymyxin B, and gentamicin. NAC alone exhibited antimicrobial and antibiofilm abilities. On the other hand, the combination of NAC and the antibiotics did not show any significant synergistic effects against P. aeruginosa.