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.

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.

Carbapenem-resistant Pseudomonas aeruginosa (CRPA) has become the leading cause of health care-associated infections. Treatment is difficult due to the lack of an effective antimicrobial therapy, and mortality is high. This study investigated the occurrence of CRPA in farm animals (buffaloes and cattle), livestock drinking water, and humans in Egypt. A total of 180 samples were examined: 50 faecal each from buffaloes and cattle, 30 of livestock drinking water, and 50 stool from humans. The samples were cultured on cetrimide agar and the plates were incubated aerobically at 37°C for 24 h. The isolates were examined for the presence of the blaKPC, blaOXA₋₄₈, and blaNDM carbapenemase-encoding genes using PCR and investigated for the exotoxin A (toxA) gene. The toxA gene from carbapenem- group resistant isolates was phylogenetically analysed. P. aeruginosa was isolated from buffaloes, cattle, drinking water, and humans, with occurrences of 40%, 34%, 10%, and 20%, respectively. Carbapenem resistance genes were found in 60%, 59%, 67%, and 70% in buffalo, cattle, water and human samples, respectively. The toxA gene was detected in 80% of samples. The phylogenetic analysis showed that cattle and water sequences were in one cluster and more related to each other than to human isolates. Occurrence of CRPA among farm animals, drinking water, and humans was high, reflecting the environmental origin of P. aeruginosa and highlighting contaminated water as a potential transmitter of CRPA to livestock and next to humans.

OBJECTIVE To evaluate histologic changes and gene expression patterns in body and limb wounds in horses in response to bacterial inoculation. SAMPLE Wound biopsy specimens from 6 horses collected on days 7, 14, 21, and 27 after excisional wounds (20 wounds/horse) were created over the metacarpal and metatarsal region and lateral thoracic region (body) and then inoculated or not inoculated on day 4 with Staphylococcus aureus and Pseudomonas aeruginosa. PROCEDURES Specimens were histologically scored for the amount of inflammation, edema, angiogenesis, fibrosis organization, and epithelialization. Quantitative PCR assays were performed to quantify gene expression of 10 inflammatory, proteolytic, fibrotic, and hypoxia-related markers involved in wound healing. RESULTS Except for gene expression of interleukin-6 on day 27 and tumor necrosis factor-α on day 14, bacterial inoculation had no significant effect on histologic scores and gene expression. Gene expression of interleukin-1β and -6, serum amyloid A, and matrix metalloproteinase-9 was higher in limb wounds versus body wounds by day 27. Gene expression of cellular communication network factor 1 was higher in limb wounds versus body wounds throughout the observation period. CONCLUSIONS AND CLINICAL RELEVANCE The lack of clear markers of wound infection in this study reflected well-known difficulties in detecting wound infections in horses. Changes consistent with protracted inflammation were evident in limb wounds, and gene expression patterns of limb wounds shared similarities with those of chronic wounds in humans. Cellular communication network factor warrants further investigation and may be useful in elucidating the mechanisms underlying poor limb wound healing in horses.

OBJECTIVE To compare antibacterial effects among 3 types of foam used with negative-pressure wound therapy (NPWT) in an ex vivo equine perfused wound model. SAMPLES Abdominal musculocutaneous flaps from 6 equine cadavers. PROCEDURES Each musculocutaneous flap was continuously perfused with saline (0.9% NaCl) solution. Four 5-cm circular wounds were created in each flap and contaminated with 106 CFUs of both Pseudomonas aeruginosa and methicillin-resistant Staphylococcus aureus (MRSA). After a 1-hour incubation period, 1 of 4 treatments (NPWT with silver-impregnated polyurethane foam [NPWT-AgPU], polyurethane foam [NPWT-PU], or polyvinyl alcohol foam [NPWT-PVA] or a nonadherent dressing containing polyhexamethylene biguanide without NPWT [control]) was randomly applied to each wound. An 8-mm punch biopsy specimen was obtained from each wound immediately before and at 6, 12, 18, and 24 hours after treatment application to determine the bacterial load for both P aeruginosa and MRSA. RESULTS The bacterial load of P aeruginosa for the NPWT-PVA treatment was significantly lower than that for the other 3 treatments at each sampling time after application, whereas the bacterial load for the NPWT-AgPU treatment was significantly lower than that for the NPWT-PU and control treatments at 12 hours after application. The bacterial load of MRSA for the NPWT-PVA treatment was significantly lower than that for the other 3 treatments at each sampling time after application. CONCLUSIONS AND CLINICAL RELEVANCE Results indicated that wounds treated with NPWT-PVA had the greatest decrease in bacterial load; however, the effect of that treatment on wound healing needs to be assessed in vivo.

Once-daily administration of aminoglycosides may be a safe and effective therapeutic regimen, on the basis of the microbiologic and pharmacokinetic characteristics of these antibiotics. This study was designed to determine serum and tissue concentrations following IV administration of gentamicin, at dosages of 6.6 mg/kg of body weight, every 24 hours, and 2.2 mg/kg, every 8 hours, for 10 days in adult horses. Nephrotoxicosis from these dosage regimens also was compared, and microbiologic effects, including postantibiotic effects, were determined with various concentrations of gentamicin against an equine clinical isolate of Pseudomonas aeruginosa. Treatment at the 6.6-mg/kg dosage resulted in maximal serum concentrations (77.93 +/- 19.90 micrograms/ml, mean +/- SEM) and area under the concentration-vs-time curves (83.79 +/- 14.97 micrograms.h/ml) that were significantly (P < 0.05) greater than those following treatment at the 2.2-mg/kg dosage (5.05 +/- 0.50 micrograms/ml and 6.03 +/- 0.66 micrograms.h/ml, respectively). Nephrotoxicosis was not induced with either dosage regimen, and postantibiotic effects were prolonged with a higher gentamicin concentration. This study provided evidence to support the use of once-daily gentamicin treatment in adult horses.

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.

Hemorrhagic pneumonia is an acute and fatal disease of farmed mink caused by Pseudomonas aeruginosa. The pathogenesis of this disease has not yet been resolved. Mink are the only animals known to be susceptible to acute, contagious, and fatal lung infections caused by P. aeruginosa. The purpose of this study was to investigate the correlation between dose-response and season of infection and to clarify whether Danish mink are carriers of P. aeruginosa on their nasal mucosa during the season for hemorrhagic pneumonia. To elucidate the pathogenesis of the disease, an infectious dose-response trial was carried out on adult mink and mink kits, both in the season for hemorrhagic pneumonia (November) as well as out of season (July). It proved difficult to infect mink via the intra-nasal route. Only 4 out of 60 infected mink developed clinical disease and were euthanized, all of them in November, illustrating that predisposing factors in the mink itself and not infectious dose might be crucial for disease development. We were able to culture P. aeruginosa from the nasal cavity of the clinically healthy experimental mink 8 d after inoculation. This indicated that the mink can carry P. aeruginosa on their nasal mucosa without developing the disease. It was not possible, however, to culture P. aeruginosa from the nasal cavity of clinically healthy mink obtained from farms in November, which indicates that the organism is not a normal part of the nasal mucosal flora of mink.

Objective—To isolate and characterize bacteriophages with strong in vitro lytic activity against various pathogenic Pseudomonas aeruginosa strains isolated from dogs with ocular infections. Sample—26 genetically distinct P aeruginosa isolates. Procedures—P aeruginosa strains were derived from dogs with naturally acquired ulcerative keratitis. From a large-scale screening for bacteriophages with potential therapeutic benefit against canine ocular infections, 2 bacteriophages (P2S2 and P5U5) were selected; host ranges were determined, and phage nucleic acid type and genetic profile were identified via enzymatic digestion. Electron microscopy was used to characterize bacteriophage ultrastructure. Bacteriophage temperature and pH stabilities were assessed by use of double-layer agar overlay titration. A cocultivation assay was used to evaluate the effect of the bacteriophages on bacterial host growth. Results—P5U5 was active against all P aeruginosa isolates, whereas P2S2 formed lytic plaques on plates of 21 (80.8%) isolates. For each bacteriophage, the genomic nucleic acid was DNA; each was genetically distinct. Ultrastructurally, P2S2 and P5U5 appeared likely to belong to the Podoviridae and Siphoviridae families, respectively. The bacteriophages were stable within a pH range of 4 to 12; however, titers of both bacteriophages decreased following heating for 10 to 50 minutes at 45° or 60°C. Growth of each P aeruginosa isolate was significantly inhibited in coculture with P2S2 or P5U5; the dose response was related to the plaque-forming unit-to-CFU ratios. Conclusions and Clinical Relevance—Bacteriophages P2S2 and P5U5 appear to be good candidates for phage treatment of infection caused by pathogenic P aeruginosa in dogs.

Objective-To evaluate the in vitro activity of an ear rinse (ER) containing tromethamine, EDTA, and benzyl alcohol on bacterial pathogens from dogs with otitis. Sample Population-Organisms were collected from ear swab specimens from the external and middle ear and included Staphylococcus spp (n = 11; Staphylococcus intermedius 7 and Staphylococcus spp 4), Pseudomonas aeruginosa (5), Proteus spp (5), beta-hemolytic streptococcus (11), and 1 control strain of each organism. Procedures-3 test solutions were evaluated including EDTA, tromethamine, and benzyl alcohol (ER); EDTA and tromethamine (ER without benzyl alcohol ER - BA); and purified water. Ten-milliliter aliquots of each test solution were transferred into 36 tubes and inoculated with one of the organisms. Samples were retrieved from each tube at 0, 15, 30, 45, and 60 minutes, transferred to Petri dishes, mixed with soybean-casein digest agar, and incubated. After incubation, plates were examined for growth, and the number of colonies was expressed as CFU per milliliter. Results-ER significantly decreased bacterial growth in vitro of P aeruginosa and beta-hemolytic streptococcal organisms within 15 minutes, Proteus spp within 30 minutes, and Staphylococcus spp within 60 minutes. Comparatively, the presence of benzyl alcohol in ER significantly decreased bacterial growth of beta-hemolytic streptococcus and Proteus spp. Conclusions and Clinical Relevance-On the basis of results of this study, future studies should be performed to evaluate the in vivo efficacy of ER alone as a treatment for otic infections caused by beta-hemolytic streptococcus, P aeruginosa, and Proteus spp and of ER combined with an antimicrobial agent for otic infections caused by Staphylococcus spp.