Spontaneous nanoemulsification of cinnamon essential oil: Formulation, characterization, and antibacterial and antibiofilm activity against fish spoilage caused by Serratia rubidaea BFMO8.

The contemporary food industry's uses of nanoemulsions (NEs) include food processing, effective nutraceutical delivery, the development of functional chemicals, and the synthesis of natural preservatives, such as phytocompounds. Although cinnamon essential oil (CEO) is widely used in the cosmetic, pharmaceutical, and food industries, it is difficult to add to aqueous-based food formulations due to its weak stability and poor water solubility. This study describes the formulation of a CEO nanoemulsion (CEONE) by spontaneous emulsification and evaluates its antibacterial and antibiofilm properties against biofilm-forming Serratia rubidaea BFMO8 isolated from spoiled emperor fish (Lethrinus miniatus). Bacteria causing spoilage in emperor fish were isolated and identified as S. rubidaea using common morphological, cultural, and 16S RNA sequencing methods, and their ability to form biofilms and their susceptibility to CEONE were assessed using biofilm-specific methods. The spontaneous emulsification formulation of CEONE was accomplished using water and Tween 20 surfactant by manipulating organic and aqueous phase interface properties and controlling particle growth by capping surfactant increases. The best emulsification, with highly stable nano-size droplets, was accomplished at 750 rpm and a 1:3 ratio concentration. The stable CEONE droplet size, polydispersity index, and zeta potential values were 204.8 nm, 0.115, and -6.05 mV, respectively. FTIR and high-resolution liquid chromatography-mass spectrometry (HR-LCMS) analyses have revealed carboxyl, carbonyl, and phenol-like primary phytochemical functional groups in CEO and CEONE, which contribute to their antibacterial and antibiofilm properties.

ShlA toxin of Serratia induces P2Y2- and α5ß1-dependent autophagy and bacterial clearance from host cells.

Serratia marcescens is an opportunistic human pathogen involved in antibiotic-resistant hospital acquired infections. Upon contact with the host epithelial cell and prior to internalization, Serratia induces an early autophagic response that is entirely dependent on the ShlA toxin. Once Serratia invades the eukaryotic cell and multiples inside an intracellular vacuole, ShlA expression also promotes an exocytic event that allows bacterial egress from the host cell without compromising its integrity. Several toxins, including ShlA, were shown to induce ATP efflux from eukaryotic cells. Here, we demonstrate that ShlA triggered a nonlytic release of ATP from Chinese hamster ovary (CHO) cells. Enzymatic removal of accumulated extracellular ATP (eATP) or pharmacological blockage of the eATP-P2Y2 purinergic receptor inhibited the ShlA-promoted autophagic response in CHO cells. Despite the intrinsic ecto-ATPase activity of CHO cells, the effective concentration and kinetic profile of eATP was consistent with the established affinity of the P2Y2 receptor and the known kinetics of autophagy induction. Moreover, eATP removal or P2Y2 receptor inhibition also suppressed the ShlA-induced exocytic expulsion of the bacteria from the host cell. Blocking α5ß1 integrin highly inhibited ShlA-dependent autophagy, a result consistent with α5ß1 transactivation by the P2Y2 receptor. In sum, eATP operates as the key signaling molecule that allows the eukaryotic cell to detect the challenge imposed by the contact with the ShlA toxin. Stimulation of P2Y2-dependent pathways evokes the activation of a defensive response to counteract cell damage and promotes the nonlytic clearance of the pathogen from the infected cell.

Effects of oil substrate supplementation on production of prodigiosin by Serratia nematodiphila for dye-sensitized solar cell.

Bacterial pigments are potential substitute of chemical photosensitizer for dye-sensitized solar cell (DSSC) due to its non-toxic property and cost-effective production from microbial fermentation. Serratia nematodiphila YO1 was isolated from waterfall in Malaysia and identified using 16S ribosomal RNA. Characterization of the red pigment produced by the bacteria has confirmed the pigment as prodigiosin. Prodigiosin was produced from the fermentation of the bacteria in the presence of different oil substrates. Palm oil exhibited the best performance of cell growth and equivalent prodigiosin yield compared to olive oil and peanut oil. Prodigiosin produced with palm oil supplementation was 93 mg/l compared to 7.8 mg/l produced without supplementation, which recorded 11.9 times improvement. Specific growth rate of the cells improved 1.4 times when palm oil was supplemented in the medium. The prodigiosin pigment produced showed comparable performance as a DSSC sensitizer by displaying an open circuit voltage of 336.1 mV and a maximum short circuit current of 0.098 mV/cm2. This study stands a novelty in proving that the production of prodigiosin is favorable in the presence of palm oil substrate with high saturated fat content, which has not been studied before. This is also among the first bacterial prodigiosin tested as photosensitizer for DSSC application.

Toxicity effects of Eriocephalus africanus L. leaf essential oil against some molecularly identified phytopathogenic bacterial strains.

Essential oil (EO) from Eriocephalus africanus L. leaves was evaluated against the growth of some phytopathogenic bacteria including Agrobacerium tumifaciens, Dickeya solani, Erwinia amylovora, Pseudomonas cichorii and Serratia pulmithica using the disc diffusion method and minimum inhibitory concentration (MIC) evaluation. Ten compounds in the EO with dominance of Artemisia ketone (2,5,5-trimethyl-2,6-heptadien-4-one) (77.92%) and ledol (19.92%) were revealed. The antibacterial activity indicated efficacy of essential oil against majority of strains isolated. The most effective action was recorded against D. solani, by 7.5 and 10 µL of oil, with 18.33 mm and 100 µg/mL as zone inhibition and MIC, respectively, whereas the lowest activity was exhibited against P. cichorii (diameter inhibition = 6.66 mm at 10 µL of oil, MIC = 100 µg/mL). The strain S. pulmithica appears to be resistant to the oil when the activity is measured by 10 µL of oil but its growth inhibition was reported with a MIC of 100 µg/mL.

The Opposite Effects of ROCK and Src Kinase Inhibitors on Susceptibility of Eukaryotic Cells to Invasion by Bacteria Serratia grimesii.

Bacterial internalization into eukaryotic cells is ensured by a sophisticated interplay of bacterial and host cell factors. Being a part of cell environment, opportunistic intracellular bacteria have developed various mechanisms providing their interaction with cell surface receptors (E-cadherin, integrins, epidermal growth factor receptor), activation of components of eukaryotic signaling pathways, and facilitation of bacterial uptake, survival, and intracellular replication. Our previous studies on the mechanisms underlying penetration of the opportunistic bacteria Serratia grimesii into cultured eukaryotic cells have shown that pretreatment of the cells with N-acetylcysteine (NAC) promotes S. grimesii invasion, and this effect correlates with the upregulation of E-cadherin expression. Since NAC has been shown to regulate expression of both Src kinase and ROCK, the aim of this work was to reveal the role of these kinases in S. grimesii invasion. We demonstrated that Y-27632, a specific inhibitor of ROCK, significantly promoted invasion of cultured eukaryotic cells by S. grimesii. On the other hand, invasion of the same cells by S. grimesii was inhibited with the Src kinase inhibitor Src-I1 and siRNA directed against RhoA. The effects of the inhibitors correlated with the corresponding changes in the E-cadherin gene expression, upregulation by the ROCK inhibition and downregulation by the Src kinase inhibition. These results prove the participation of ROCK and Src protein kinases in the invasion of eukaryotic cells by the opportunistic pathogen S. grimesii, as well as suggest that other signaling pathways might be involved in S. grimesii uptake, that are promoted by the ROCK inhibition with Y-27632.

Endophytic bacteria isolated from Solanum nigrum L., alleviate cadmium (Cd) stress response by their antioxidant potentials, including SOD synthesis by sodA gene.

Cadmium (Cd) is a toxic heavy metal and an abiotic stressor to plants; however, inoculation of endophytic bacteria can raise resistance in plants against Cd, as well as improve plant growth. In the present study, two endophytic bacterial strains were isolated from Solanum nigrum, identified as Serratia sp. IU01 and Enterobacter sp. IU02 by 16S DNA sequencing. Both IU01 and IU02 were tolerant up to 9.0 mM of Cd in culture broth and successive increase in Cd concentration from 0 mM to 9.0 mM, led to an increase in the SOD enzyme activity of the isolates. Both strains were capable of indole-3-acetic acid (IAA) synthesis and phosphate solubilization, detected through gas spectrometry-mass chromatography (GC-MS) and Pikovskaya agar medium respectively. Brassica juncea plants stressed with 0-25 mg/kg Cd showed retardation in all growth attributes, however, inoculation of strain IU01 and IU02 significantly promoted the plant growth attributes as compared to control. Moreover, antioxidant enzymes and metabolites against reactive oxygen species (ROS) including polyphenol oxidase (PPO), peroxidase (POD), catalase (CAT), superoxide dismutase (SOD), alcohol dehydrogenase (ADH), reduced glutathione (GSH), malondialdehyde (MDA), flavonoid and polyphenolic contents were also significantly relieved by inoculation of IU01 and IU02 in plant exposed to different concentration of Cd stress as compared to control plants. Phytohormone production, phosphate solubilization, and/or antioxidative support of IU01 and IU02 might be responsible for growth promotion and Cd resistance in the plant.

Metabolic response of bacteria to elevated concentrations of glyphosate-based herbicide.

Glyphosate-based herbicides (GBHs) are the most widespread commonly used broad-spectrum herbicides that contaminate soils and waters, are toxic to bacteria, plants and animals, and have been classified as 'probably carcinogenic to humans' by the International Agency for Research on Cancer in 2015. Particular soil bacteria and fungi can degrade GBHs, hence, search for new GBH-degrading strains or microbial consortia, effective under specific growth conditions and local environment, seems to be a promising solution for bio-remediation of glyphosate-contaminated environment. Consequently, there is a need for rapid and informative methods to evaluate the GBH-induced changes of the metabolic pathways in cells, that may serve as indicators of GBH-degrading potential. Three new GBH-degrading bacterial strains, Pseudomonas sp., Actinobacteria and Serratia sp. were isolated from sludge of municipal waste water treatment plant (Daugavgriva, Riga, Latvia), agricultural soil and plant tissue, respectively. This study examined the response of these isolates to elevated concentrations of glyphosate (GLP) (100 and 500 mg/L) in GBH Klinik® 360 SL. The GBH-induced shift of metabolic activity in cells of Pseudomonas sp. was shown by tests on EcoPlates™. Fourier transform infrared (FTIR) spectroscopy analyses were used to evaluate the metabolomic response of bacteria to elevated concentrations of GBH in the growth environment. The spectra of Pseudomonas sp. and Serratia sp., incubated with and without GBH, were similar, thus indicating their GBH-resistance. The absorption at 1736 cm-1, assigned to ester carbonyl stretch vibrations, was detected in spectra of all three bacteria. The highest ester content was detected in Actinobacteria grown in medium with 1.0% molasses and 100 or 500 mg/L GLP in GBH Klinik®. An increase of cellular amounts of esters, either those of phospholipids or poly-ß-hydroxybutyrates, indicates degradation of GLP. Therefore, monitoring the ester carbonyl stretch vibration band in FTIR spectra of bacterial biomass may speed up the search GBH-degrading strains. Microbiological tests and cell metabolic response studies by FTIR spectroscopy showed that the three new isolates of Pseudomonas sp., Actinobacteria and Serratia sp. were resistant to elevated concentrations of GBH Klinik® in growth environment and exhibited the potential for GBH degradation.

Cadmium-tolerant bacteria induce metal stress tolerance in cereals.

Cadmium usually hampers plant growth, but bacterial inoculation may improve stress tolerance in plants to Cd by involving various mechanisms. The objective was to characterize and identify bacteria that improve plant growth under Cd stress and reduce Cd uptake. Cadmium-tolerant bacteria were isolated from rhizosphere soil, which was irrigated with tannery effluent, and six strains were selected as highly tolerant to Cd, showing minimum inhibitory concentration as 500 mg L(-1) or 4.45 mmol L(-1). These strains were identified by 16S rRNA gene analysis and functional analysis in regard to plant growth promotion characteristics. To determine their effect on cereal growth under Cd stress, seeds were inoculated with these strains individually and grown in soil contaminated with three Cd levels (0, 40 and 80 mg kg(-1)). Biomass production, relative water content (RWC), electrolyte leakage (ELL) and tissue Cd concentration were measured. Biomass of both cereals was inhibited strongly when exposed to Cd; however, bacterial inoculation significantly reduced the suppressive effect of Cd on cereal growth and physiology. The bacterial isolates belonged to the genera Klebsiella, Stenotrophomonas, Bacillus and Serratia. Maize was more sensitive than wheat to Cd. Klebsiella sp. strain CIK-502 had the most pronounced effects in promoting maize and wheat growth and lowering Cd uptake under Cd stress.

Serratia entomophila bet gene induction and the impact of glycine betaine accumulation on desiccation tolerance.

AIMS: The genes involved in choline transport and oxidation to glycine betaine in the biopesticidal bacterium Serratia entomophila were characterized, and the potential of osmoprotectants, coupled with increased NaCl concentrations, to improve the desiccation tolerance of this species was investigated. METHODS AND RESULTS: Serratia entomophila carries sequences similar to the Escherichia coli betTIBA genes encoding a choline transporter and dehydrogenase, a betaine aldehyde dehydrogenase and a regulatory protein. Disruption of betA abolished the ability of Ser. entomophila to utilize choline as a carbon source. Quantitative reverse-transcriptase PCR analysis revealed that betA transcription was reduced compared to that of the upstream genes in the operon, and that NaCl and choline induced bet gene expression. Glycine betaine and choline increased the NaCl tolerance of Ser. entomophila, and osmotically preconditioned cultures survived better than control cultures following desiccation and immediately after application to agricultural soil. CONCLUSIONS: Addition of glycine betaine and NaCl to growth medium can greatly enhance the desiccation survival of Ser. entomophila, and its initial survival in soil. SIGNIFICANCE AND IMPACT OF THE STUDY: Serratia entomophila is sensitive to desiccation and does not persist under low soil moisture conditions. Techniques described here for enhancing the desiccation survival of Ser. entomophila can be used to improve formulations of this bacterium, and allow its application under a wider range of environmental conditions.

Effect of tea polyphenols on microbiological and biochemical quality of Collichthys fish ball.

BACKGROUND: Tea polyphenols (TP), as the most active constituents of tea, are considered natural food additives. This study examined the preservative properties of TP for Collichthys fish ball in well storage. Vacuum-packed Collichthys fish balls were treated with 0, 0.1, 0.15, 0.20, 0.25, and 0.30 g kg(-1) TP and stored at 0 °C for 17 days. RESULTS: Microbiological results were obtained using a biochemical test, API system kit, and 16S rDNA sequence analysis. Results confirmed that the dominant bacteria in Collichthys fish balls are the genera Serratia and Pseudomonas. Total viable counts dropped two orders of magnitude in Collichthys fish balls with 0.25 g kg(-1) TP compared with the control. The advantages of total volatile basic nitrogen value, 2-thiobarbituric acid value and texture value were clearly observed, whereas pH and whiteness value exhibited no significant decrease for the group treated with 0.25 g kg(-1) TP. More than 0.25 g kg(-1) TP added could retain excellent fish ball characteristics in terms of sensory assessment after 17 days. CONCLUSION: The shelf life of Collichthys fish balls supplemented with tea polyphenols can be prolonged for an additional 6 days in good condition at 0 °C storage.

2-amino-3-(oxirane-2,3-dicarboxamido)-propanoyl-valine, an effective peptide antibiotic from the epiphyte Pantoea agglomerans 48b/90.

The epiphyte Pantoea agglomerans 48b/90, which has been isolated from soybean leaves, belongs to the Enterobacteriaceae, as does the plant pathogen Erwinia amylovora, which causes fire blight on rosaceous plants such as apples and leads to severe economic losses. Since P. agglomerans efficiently antagonizes phytopathogenic bacteria, the P. agglomerans strain C9-1 is used as a biocontrol agent (BlightBan C9-1). Here we describe the bioassay-guided isolation of a peptide antibiotic that is highly active against the plant pathogen E. amylovora and pathovars of Pseudomonas syringae, and we elucidate its structure. Bioassay-guided fractionation using anion-exchange chromatography followed by hydrophobic interaction liquid chromatography yielded the bioactive, highly polar antibiotic. The compound was identified as 2-amino-3-(oxirane-2,3-dicarboxamido)-propanoyl-valine by using high-resolution electrospray ionization mass spectrometry and nuclear magnetic resonance techniques. This peptide was found to be produced by three of the nine P. agglomerans strains analyzed. Notably, the biocontrol strain P. agglomerans C9-1 also produces 2-amino-3-(oxirane-2,3-dicarboxamido)-propanoyl-valine. Previously, 2-amino-3-(oxirane-2,3-dicarboxamido)-propanoyl-valine has been characterized only from Serratia plymuthica. 2-Amino-3-(oxirane-2,3-dicarboxamido)-propanoyl-valine has been shown to inhibit the growth of the human pathogen Candida albicans efficiently, but its involvement in the defense of epiphytes against phytopathogenic bacteria has not been investigated so far.

Contaminação de halos doadores córneo-esclerais em ceratoplastia penetrante no Hospital de Clínicas de Porto Alegre / Positive corneoscleral rim culture in penetrating keratoplasty at the Porto Alegre Clinical Hospital

OBJETIVOS: Avaliar a incidência de positividade de culturas de halos doadores córneo-esclerais preservados em Optisol GS, identificar os patógenos envolvidos, a sensibilidade dos mesmos à gentamicina e a ocorrência de infecções em olhos receptores. MÉTODOS: Foram analisadas retrospectivamente 163 culturas de halos córneo-esclerais cujos botões corneanos foram utilizados em transplantes de córnea no Hospital de Clínicas de Porto Alegre entre janeiro de 2001 e janeiro de 2003. Os halos foram divididos em dois segmentos, metade inoculada em meio Sabouroud e a outra metade em tioglicolato, com posterior semeadura em ágar-sangue, ágar-chocoloate e meio de MacConkey, conforme necessidade para identificação dos patógenos. Os prontuários dos pacientes receptores foram revisados. RESULTADOS: Dos 163 halos analisados, 11 apresentaram culturas positivas, correspondendo a 6,7 por cento do total. Destes, quatro foram por Staphylococcus epidermidis, um por Staphylococcus aureus, um por Serratia sp, um por Pseudomonas aeruginosa e os outros quatro por diferentes subtipos de Candida (dois por Candida sp, um por Candida albicans e um por Candida parapapilosis). No antibiograma, todas as bactérias apresentaram-se resistentes à gentamicina. Nenhum olho que recebeu córnea com cultura positiva apresentou infecção após a cirurgia. CONCLUSÕES: Baixos índices de positividade de cultura de halos utilizados em transplantes de córnea no Hospital de Clínicas de Porto Alegre foram encontrados. Os patógenos mais freqüentemente identificados não apresentam boa cobertura pelos antimicrobianos presentes nos meios de preservação. A cultura de halos corneanos é recomendada para auxílio no tratamento de possível infecção ocular pós-cirúrgica.

PURPOSE: To determine the incidence of positive corneoscleral rim cultures preserved in Optisol GS medium, to identify pathogens involved and possible recipient eye infection. METHODS: A hundred sixty-three corneoscleral rim cultures penetrating keratoplasties performed from January 2001 to January 2003 in the Hospital de Clínicas de Porto Alegre were reviewed. Enucleations and corneal storage were done as aseptic as possible and gentamicin 0.3 percent was instilled. Corneoscleral rim was divided into two segments, half was inoculated into Sabouraud broth and the other half into thioglycolate broth; inoculation into blood agar, chocolate agar and MacConkey agar was done later if necessary for pathogen identification. The receiver's eye data were reviewed. RESULTS: There were eleven positive cultures (6.7 percent) out of 163 evaluated corneoscleral rim cultures. Of these, four were Staphylococcus epidermidis, one was Staphylococcus aureus, one was Serratia sp., one was Pseudomonas aeruginosa and the other four were different subtypes of Candida (two Candida sp., one Candida albicans and one Candida parapapilosis). All pathogens were resistant to gentamicin. None of the eleven cases of positive corneoscleral rim cultures resulted in ocular infection at the receiver's eyes (six months follow-up). CONCLUSIONS: We found low rates of positive corneoscleral rim cultures after penetrating keratoplasty at the Porto Alegre Clinical Hospital. The most frequent involved pathogens were Staphylococcus sp and Candida sp. Although we did not identify any postoperative infection at the receiver's eyes, we recommend corneoscleral rim culture for guidance of postoperative infection, a rare but possible devastating ocular event.

Fluctuation in recoverable nickel and zinc resistant copiotrophic bacteria explained by the varying zinc ion content of Torsa River in different months.

Heavy metal content analysis of River Torsa in India did not indicate any alarming level of toxicity for human consumption but revealed variation at the ppb level in different months. The variation in recoverable nickel and zinc resistant copiotrophic (or eutrophic) bacterial counts was explained by the variation of the zinc content (34.0-691.3 ppb) of the river water in different sampling months. Growth studies conducted with some purified nickel and/or zinc resistant strains revealed that pre-exposure of the cells to ppb levels of Zn(2+), comparable to the indigenous zinc ion concentration of the river, could induce the nickel or zinc resistance. A minimum concentration of 5-10 microM Zn(2+ )(325-650 ppb) was found effective in inducing the Nickel resistance of the isolates. Zinc resistance of the isolates was tested by pre-exposing the cells to 4 microM Zn(2+ )(260 ppb). The lag phase was reduced by 6-8 h in all the cases. Biochemical characteristics and phylogenetic analysis based on 16S rDNA sequence indicated that some of the Torsa River isolates, having inducible nickel and zinc resistance, are members of the genus Pseudomonas, Acinetobacter, Bacillus, Enterobacter, Serratia and Moraxella.

Use of several inducer and substrate antibiotic combinations in a disk approximation assay format to screen for AmpC induction in patient isolates of Pseudomonas aeruginosa, Enterobacter spp., Citrobacter spp., and Serratia spp.

Two-hundred consecutive, single patient isolates of Enterobacter spp., Serratia spp., Citrobacter spp., and Pseudomonas aeruginosa were evaluated for AmpC production using a variety of inducer-substrate antibiotic combinations in a disk approximation format. The combinations examined included cefoxitin-piperacillin, imipenem-cefotaxime, imipenem-ceftazidime, imipenem-piperacillin-tazobactam, and imipenem-cefoxitin. All isolates were also screened for the presence of extended-spectrum beta-lactamase (ESBL) activity. In total, 85.5% of isolates were shown to be inducible for the production of AmpC by one or more inducer/substrate combinations and 11% of all isolates were stably derepressed for the expression of AmpC. Of all of the combinations, imipenem/piperacillin-tazobactam provided the greatest sensitivity (97.1%). All combinations were 100% specific when a positive test was observed. Given this background among these organisms in our institution, it is reasonable to develop an antibiotic reporting strategy that favors the selection of agents for therapy of these organisms that do not serve as labile substrates of AmpC.

Absorption of ester prodrugs in Caco-2 and rat intestine models.

The aim of this study was to elucidate the absorption mechanism in Caco-2 and rat intestine models in order to improve the accuracy of prediction of oral absorption of ester prodrugs. Pivampicillin and cefcapene pivoxil hydrochloride (CFPN-PI), ester-type oral antibiotics, were chosen as model ester prodrugs. The level of esterase activity in Caco-2 cells was lower than that measured in the rat jejunum when p-nitrophenyl acetate was used as a substrate. Almost complete ester hydrolysis occurred before the ester prodrugs reached the basolateral side of the monolayer, and the disappearance of prodrugs was thought to be due to metabolism or transport after addition to the apical side of the monolayer. When pivampicillin and CFPN-PI were used, the amounts of ampicillin and cefcapene (CFPN) produced by hydrolysis of prodrugs were increased because intracellular degradation of prodrugs resulted in intracellular accumulation. On the other hand, when ampicillin or CFPN was used, only a small amount of the drug reached the basolateral side of the monolayers and no intracellular accumulation was observed. The permeability of CFPN-PI, the solubility of which is dependent on the acidity of gastric juice, across a Caco-2 monolayer or rat intestine, was also investigated by using an in vitro system that mimics the physiological state of the human gastrointestinal tract. The oral absorption of CFPN-PI in humans is predicted to be good either in the Caco-2 model or in the rat intestine model. It is concluded that our system may be a valuable tool for evaluation of oral absorption of ester prodrugs metabolized during permeation through the intestinal epithelium. Broader evaluation of such a system is warranted.

Report of six cases of human infection by Serratia plymuthica.

Serratia plymuthica is an uncommon cause of human infection. Only one case of chronic osteomyelitis and two cases of sepsis secondary to central venous catheter infection have been documented. We report the isolation of S. plymuthica from six patients. The organism was recovered from blood cultures in three cases in which the patients had lymphoblastic leukemia, lymphoma, or stroke. Two isolates were recovered from exudates (following knee and abdominal surgery). In the last case, the organism was isolated from the peritoneal fluid of a patient with cholecystitis. The infection was considered nosocomial in five cases and community acquired in the other.

[Opportunistic infection and systemic dissemination in burns].

A total of 303 strains of opportunistic bacteria were isolated from our burned patients during April, 1980 to December, 1987. Among which Pseudomonas aeruginosa accounted for 161 strains (54.1%), Serratia accounted for 56 strains (18.5%), just the two species accounted for 72.6% in total. Among twenty commonly used antibiotics, Amikacin and Polymyxin-B were comparatively sensitive. Further reviewing the drug sensitivity of the two species, we found the sensitivity rates were variable among the strains isolated from different sources. To Polymyxin-B, strains isolated from wound surfaces were of 86.1% and 90.7% respectively, from subeschar tissues or visceral organs were of 53.8% and 39.3%, from blood stream were of 44.4% and 40%. It seemed that the drug resistance of the invading organisms was stronger than that of surface ones. It suggested that the Pseudomonas aeruginosa and Serratia play an important role in infection of burned patients.

[Plasmids and genetic determinants of antibiotic resistance of gram-negative bacteria]. / Plazmidy i geneticheskie determinanty rezistentnosti k antibiotikam gramotritsatel'nykh bakterii.

Distribution of plasmids and genetic determinants of antibiotic resistance was studied in 129 strains of Pseudomonas, Klebsiella, Serratia and Enterobacter isolated from oncological patients. It was shown that 56 isolates contained the plasmids, 9 conjugative plasmids being plasmids with broad bacterial host spectrum. A significant part of the strains contained genes controlling production of APH (3"), type II APH (3'), type I and II DHPS and type type II DHFR. Genetic determinants of tetracycline resistance of classes D and E were detected for the first time in the strains of Klebsiella, Serratia and Pseudomonas aeruginosa.

Resistance surveillance programs and the incidence of gram-negative bacillary resistance to amikacin from 1967 to 1985.

Data relating to amikacin resistance among gram-negative bacilli were obtained by means of a review of published literature and resistance surveillance studies. Data from the first several years of amikacin use are difficult to interpret because the 10-micrograms disk used for Kirby-Bauer susceptibility testing resulted in apparent greater resistance than the present 30-micrograms disk. A large United States susceptibility surveillance program that monitors antibiotic use has shown a trend since 1977 of greater susceptibility of Serratia species and greater resistance among Pseudomonas aeruginosa for all the aminoglycosides. Pseudomonas resistance to amikacin has shown the smallest increase of any aminoglycoside. Several hospitals (Strong Memorial Hospital, University of Maryland Cancer Center, and Minneapolis Veterans Administration Medical Center) have reported either no significant change or a decrease in resistance to amikacin when it was the most frequently used aminoglycoside. In a large, 14-center, prospective study, high-level use of amikacin resulted in a significant decrease in resistance to gentamicin and tobramycin (p less than 0.01) and a marginal increase (p less than 0.05) in amikacin resistance. Significantly increased amikacin resistance has been reported from two institutions, neither of which used amikacin as the predominant aminoglycoside. Overall, the high-level use of amikacin in large multi-center surveillance programs for as long as five years has not resulted in a significant increase in amikacin resistance rates at any of the individual institutions surveyed.

The osmoprotective properties of urine for bacteria: the protective effect of betaine and human urine against low pH and high concentrations of electrolytes, sugars, and urea.

Growth of Escherichia coli was inhibited in a defined minimal medium by high concentrations of electrolytes and sugars in direct relation to their osmotic strength. Choline, betaine, proline, and human urine increased resistance to these substances. In contrast, the toxic effect of urea was not altered directly by betaine or urine, but was reduced in the presence of other osmolytes. The osmolyte protective effect was augmented by betaine. The osmoprotective effect of betaine and urine was confirmed with 40 strains of enteric bacteria. Urine from 19 healthy subjects contained osmoprotective activity greater than that observed with betaine. A methanol extract of urine was found to be highly protective. Although betaine was present in the extract, it could not account for all the protective activity. Urine contains additional low-molecular-weight osmoprotective agents.