Biocide analysis, usability and stability

Biosidien toimittajat tavallisesti suorittavat biosidien annostelumäärien hallinnan paperi- ja kartonkiteollisuudessa. Useimmiten annostelun hallinta määritetään epäsuorilla menetelmillä, kuten esimerkiksi määrittämällä bakteerien kasvua. Biosidien tehoaineiden todellista konsentraatiota tai määrää prosessivesissä tai lopputuotteessa ei tavallisesti mitata. Diplomityössä kehitettiin kolmelle paperiteollisuudessa yleisesti käytetyllä biosidin tehoaineelle analyyttiset menetelmät. Menetelmät kehitettiin glutaraldehydille, 2,2-dibromi-3-nitriilipropionamidi:lle (DBNPA) ja 5-kloori-2-metyyli-4-isotiatsoliini-3-oni:lle (CMI). Kehitettyjä menetelmiä käytettiin tehoaineiden stabiilisuuden seuraamiseen vesiliuoksessa eri pH:ssa ja lämpötilassa. Lisäksi kartonkinäytteistä tehtiin uuttokokeita ja yritettiin kehittää uuttomenetelmät, joilla pystyttäisiin määrittämään biosidien tehoaineiden jäännöspitoisuuksia lopputuotteesta. Glutaraldehydille ja CMI:lle onnistuttiin kehittämään uuttomenetelmät, joilla pystyttiin määrittämään kartongista tutkittujen tehoaineiden jäännöspitoisuudet. Saadut tulokset vaikuttavat realistisilta. Glutaraldehydille ja DBNPA:lle tehtiin stabiilisuuskokeita ja tulokset ovat samankaltaisia mitä muut tutkijat ovat saaneet.

Presença de compostos carbonílicos no ar em ambientes internos na cidade de São Paulo

The presence of low carbonyl compounds (C1-C4) and glutaraldehyde was observed in 14 work-locations in São Paulo city, Brazil, during January to July 1997 period. The quantification of other carbonyls was not possible due to a sampling artifact caused probably by undesirable reactions between the ambient ozone and the organic substract of C18 cartridge used to collect carbonyls. High indoor concentrations, compared to outdoor levels, were observed for all compounds. Formaldehyde was the most abundant species (29 ppb) followed by acetone and acetaldehyde (25 and 17 ppb) with a small contribution of propanal, crotonaldehyde and C4 isomers (0.7 to 1.5 ppb) when related to the total indoor carbonyls level. Glutaraldehyde was observed only in two different occupational locations in a hospital at high concentrations (121 ppb). In general, high individual carbonyl levels as well as total carbonyls levels found in several locations indicated an unpropitious air quality for the occupants of these non-industrial sites.

Estudo da influência da natureza de reticulantes e aditivos orgânicos sobre o comportamento de géis de quitosana

The physical-chemical process of swelling in water-based gel of natural polymers is investigated with the purpose of applying these systems to biomedical materials for controlled release of drugs. In this work we develop a study about the sol-gel transition of solutions of chitosan in the presence of formaldehyde and glutaraldehyde like crosslinking agents and we have determined the effect of many aditives in the time of gelification from the elaborated sistems. The phisical-chemistry process of swelling of the formed gels was evaluated in function of the degree of crosslinking of the incorporated aditives and the pH. Gelling times of chitosan solutions were obtained using viscosimetric measurement, in the pre-gel state, as well as condutivity ones.The results obtained suggest that component concentration modifies the kinetic profile of the transition and the swelling behavior. Regarding H+ content, the gels were highly susceptible to swelling in acidic conditions, which characterize this system as pH - sensitive.

Construção e aplicação de biossensores usando diferentes procedimentos de imobilização da peroxidase de vegetal em matriz de quitosana

Biosensors were developed by immobilization of gilo (Solanum gilo) enzymatic extract on chitosan biopolymers using three different procedures: glutaraldehyde, carbodiimide/glutaraldehyde and epichlorohydrin/glutaraldehyde. The best biosensor performance was obtained after the immobilization of peroxidase on chitosan with epichlorohydrin/glutaraldehyde. Linear analytical curves for hydroquinone concentrations from 2.5x10-4 to 4.5x10-3 mol L-1 with a detection limit of 2.0x10-6 mol L-1 and recovery of hydroquinone ranging from 95.1 to 105% were obtained. The relative standard deviation was < 1.0 % for a solution of 3.0x10-4 mol L-1 hydroquinone and 2.0x10-3 mol L-1 hydrogen peroxide in 0.1 mol L-1 phosphate buffer solution at pH 7.0 (n=8). The lifetime of this biosensor was 6 months (at least 300 determinations).

Biossensor enzimático para detecção de fungicidas ditiocarbamatos: estudo cinético da enzima aldeído desidrogenase e otimização do biossensor

Initially, all major factors that affect the rate of the AldH-catalyzed reaction (enzyme concentration, substrate concentration, temperature and pH) were investigated. Optimal activity was observed between pH values of 7.5 and 9.5 in the temperature range of 25 to 50 ºC. Kinetic parameters, such as Km (2.92 µmol L-1) and Vmax (1.33 10-2 µmol min-1) demonstrate a strong enzyme-substrate affinity. The sensors were based on screen-printed electrodes modified with the Meldola Blue-Reinecke salt (MBRS) combination. Operational conditions (NAD+ and substrate contents, enzyme loading and response time) were optimized. Also, two enzyme immobilization procedures were tested: entrapment in poly(vinyl alcohol) bearing styrylpyridinium groups (PVA-SbQ) and crosslinking with glutaraldehyde. Chronoamperometry was employed to observe the biosensor responses during enzymatic hydrolysis of propionaldehyde and also to construct inhibition curves with maneb and zineb fungicides. Best results were found with the following conditions: [NAD+] = 0.25 mmol L-1; [propionaldehyde] = 80 µmol L-1; enzyme loading = 0.8 U per electrode; response time = 10 min, and inhibition time = 10 min. Current intensities around 103 ± 13 nA with the sensors and good stability was obtained for both immobilization procedures. Detection limits, calculated using 10% inhibition were 31.5 µg L-1 and 35 µg L-1 for maneb and zineb, respectively. Results obtained with other MBRS-modified electrodes consisting of mono and bi-enzymic sensors were compared. The ability to catalyze NADH oxidation by MB was also highlighted.

Microencapsulação do agente quelante sulfoxina em microesferas de quitosana preparadas por spray drying como novo adsorvente para íons metálicos

In this work, a new adsorbent was prepared by microencapsulation of sulfoxine into chitosan microspheres by the spray drying technique. The new adsorbent was characterized by Raman spectroscopy, scanning electron microscopy and microanalysis of energy dispersive X-rays. The Cu(II) adsorption was studied as a function of pH, time and concentration. The optimum pH was found to be 6.0. The kinetic and equilibrium data showed that the adsorption process followed the pseudo second-order kinetic model and the Langmuir isotherm model over the entire concentration range. An increase of 8.0% in the maximum adsorption capacity of the adsorbent (53.8 mg g-1) was observed as compared to chitosan glutaraldehyde cross-linked microspheres.

Preparação e caracterização de blendas de quitosana/poli(álcool vinílico) reticuladas quimicamente com glutaraldeído para aplicação em engenharia de tecido

In this study, novel Chitosan/PVA based films were chemically crosslinked by glutaraldehyde, under pH=(4,00 ±0,05), in order to achieve structures tailored for wound tissue engineering applications. Both precursors and developed films were characterized by FTIR, SEM and XRD in order to determine the presence of chemicals groups and nanostructural order, respectively. The results have shown that the glutaraldehyde crosslinking have altered the crystallinity of pure chitosan and the increase on the C=N bands and simultaneous decrease on NH2 bands suggested that Chitosan/GA crosslinking has preference to occur in carbon-2 of the saccharide ring by the Schiff's base reaction. Also, FTIR spectroscopy clearly showed that crosslinking has also taken place with blends of PVA and chitosan. The mechanical properties presented high degree dependence with on the increase of the content of chitosan and glutaraldehyde. The results have indicated that, by controlling the ratio [PVA]/[chitosan] in the blends and the extent of chemical crosslinking, it was possible to tailor the hybrid network produced aiming to obtain properties of interest for the specific application.

Identificação de microcistina LR ao nível molecular empregando microscopia de força atômica

Microcystins are non-ribosomal peptides that must be detected for its health concern. Here, microcystin LR and its specific antibody were respectively tethered to the substrate and to the tip of an atomic force microscope, after surface functionalization using 3-aminopropyltriethoxysilane and glutaraldehyde. Functionalization was confirmed comparing topographic images taken on bare and modified tips. Force versus distance curves were successfully used to measure the specific antibody-antigen interactions comparing with a control in which microcystin was initially blocked by incubation with free antibodies. The results showed unequivocally the specific recognition of MLR, suggesting that this method could be useful for biosensor development.

Imobilização e caracterização de lacase e seu uso na biodegradação de efluentes de indústrias papeleiras

Laccase from Aspergillus sp was immobilized on glutaraldehyde-activated chitosan beads. A comparative study between free and immobilized laccase was conducted and the potential of the resulting immobilized derivative in the biodegradation of pulp and paper mill effluent was evaluated. The immobilized laccase is more resistant to various denaturing conditions, which allows for the reduction of 65% of the phenols (total and low molecular weight) and loss of 60% of total color in the effluent. These results show the potential of the immobilized laccase in the biodegradation of phenols, the chemical agents responsible for the high toxicity of the effluent generated in cellulose pulp industries.

Imobilização de lacase de Aspergillus sp. em quitosana e sua aplicação na bioconversão de fenóis em reatores de leito fixo

The immobilization of laccase on chitosan by cross-linking and application of the immobilized laccase in the bioconversion of phenolic compounds in batch and fixed bed reactors were studied. The process for immobilization of enzyme was optimized using a rotational central composite design. The optimized conditions to generate immobilized laccase with maximal activity were determined to be a glutaraldehyde concentration of 1.0% (v/v), a pH of 6.0, an immobilization time of 5.0 hours and an enzyme concentration of 5.2 g L-1. In packed bed reactors, the activity of the immobilized enzyme is maintained for a longer time in the bioconversion of 2,6-dimethoxyphenol than in the bioconversion of syringaldazine.

Amperometric biosensor for ascorbic acid

A L-ascorbic acid biosensor based on ascorbate oxidase has been developed. The enzyme was extracted from the mesocarp of cucumber (Cucumis sativus) by using 0.05 mol L-1 phosphate buffer, pH 5.8 containing 0.5 mol L-1 NaCl. After the dialysis versus phosphate buffer 0.05 mol L-1 pH 5.8, the enzyme was immobilized onto nylon net through glutaraldehyde covalent bond. The membrane was coupled to an O2 electrode and the yielding reaction monitored by oxygen depletion at -600 mV using flow injection analysis optimized to 0.1 mol L-1 phosphate buffer pH 5.8, as the carrier solution and flow-rate of 0.5 mL min-1. The ascorbic acid calibration curve was linear from 1.2x10-4 to 1.0x10-3 mol L-1. The evaluation of biosensor lifetime leads to 500 injections. Commercial pharmaceutical samples were analyzed with the proposed method and the results were compared with those obtained by high-performance liquid chromatography (HPLC).

Infecção por micobactérias de crescimento rápido após procedimentos videocirúrgicos - a hipótese do glutaraldeído

Between August 2006 and February 2007, in the state of Rio de Janeiro, Brazil, a massive outbreak of RGM infections after video laparoscopy was mainly associated to the recently described Mycobacterium massiliense species. All confirmed and probable cases reports described the use of high-level disinfection of medical devices by using 2% glutaraldehyde (2% GA) for 30 min before the surgical procedures. We investigated the susceptibility of the M. massiliense isolates recovered during the outbreak to high-level disinfection after 30 min, 1h, 6h and 10h of exposure to the commercial disinfectants. Reference strains for official mycobactericidal tests such as Mycobacterium abscessus, Mycobacterium bovis, Mycobacterium chelonae, Mycobacterium neoaurum and Mycobacterium smegmatis were included as controls. Although all the reference strains were eliminated in 30 min of exposure to 2% GA, we observed the recovery of all M. massiliense clinical isolates even after 10h of exposure. This study suggests that failures in high-level disinfection and the high tolerance of these M. massiliense clinical strains to the 2% GA were strongly associated to the magnitude of the outbreak.

Infecções por micobactérias de crescimento rápido resistentes a desinfetantes: uma problemática nacional?

Rapidly growing mycobacteria (RGM) are opportunistic microorganisms and widely distributed into aqueous environment and soil. Human RGM infections are usually associated with contaminated solutions or medical instruments used during invasive procedures. RGM postsurgical infections have recently emerged in Brazil and have caused national alert, considering the risk factors and epidemiological aspects. This study aimed at analysing the main factors linked to the recent RGM outbreaks, with focus on the national epidemic of Mycobacterium massiliense infections related to the BRA100 strains resistant to 2% glutaraldehyde commercial solutions commonly used for preoperative high-level disinfection. Based on previous studies and laboratorial results of assays and colaborations, it has been observed that the cases have been associated with videolaparoscopy for different applications and elective esthetic procedures, such as lipoaspiration and mammary prosthesis implant. Furthermore, outbreaks between 2004 and 2008 and the epidemic in Rio de Janeiro state may be considered particular Brazilian events. Although there are a few epidemiological published studies, some hypotheses based on common aspects related to most national nosocomial occurrences are possible, such as lack of protocols for cleaning and high-level disinfection, use of 2% glutaraldehyde as high-level disinfectant for surgical instruments, and dissemination of M. massiliense BRA100 by unknown mechanisms.

The materno-fetal interface in llama (Lama guanicoe glama)

Samples from 9 llamas (28 through 36 weeks of gestation) were collected and fixed in 4% buffered paraformaldehyde (light microscopy) and in 2.5% buffered glutaraldehyde (transmission and scanning electron microscopy). The material was processed in paraplast and slides (5mm) were stained with HE, PAS, Masson-Trichrome, acid phosphatase and Perl's. The uteroferrin was immunolocalized. The results show that llama placenta is chorioallantoic, diffuse, folded and epitheliochorial, and the fetus is covered with an epidermal membrane. The trophoblast cells have variable morphology: cubic, rounded and triangular cells, with cytoplasm containing PAS-positive granules. Binucleated cells with large cytoplasm and rounded nuclei, as well as giant trophoblastic cells with multiple nuclei were also observed. Numerous blood vessels were observed beneath the cells of the uterine epithelium and around the chorionic subdivided branches. Glandular activity was shown by PAS, Perl's, and acid phosphatase positive reactions in the cytoplasm and glandular lumen, and by immunolocalization of the uteroferrin in the glandular epithelium. The uterine glands open in spaces formed by the areoles, which are filled by PAS-positive material. The llama fetus was covered by the epidermal membrane, composed of stratified epithelium, with up to seven layers of mono-, bi- or trinucleated cells. The high level of maternal and fetal vascularization surfaces indicates an intense exchange of substances across both surfaces. The metabolic activity shown in the uterine glands suggests an adaptation of the gestation to the high altitudes of the natural habitat of this species.

Ultrasuodatusmembraanin modifiointi entsyymeillä suodatusominaisuuksien parantamiseksi

Tämän diplomityön tarkoituksena oli kiinnittää lakkaasientsyymi polyeetterisulfonimembraaniin suodatusominaisuuksien parantamiseksi. Lakkaasientsyymin tiedetään pilkkovan ligniiniä ja kiinnittämällä lakkaasientsyymi membraaniin tavoiteltiin ligniinin aiheuttaman membraanin likaantumisen vähentämistä. Tällöin vältyttäisiin lisäksi erilliseltä esikäsittely vaiheelta ja voitaisiin saada puhtaampi lopputuote. Lakkaasientsyymivalmisteena tutkimuksessa käytettiin Novozym® 51003 ja ristisilloittajana käytettiin glutaarialdehydiä. Vapaan ja kiinnitetyn lakkaasientsyymin aktiivisuuden määritettiin 2,2-atso-bis(3- etyylibentsotiatsolyyli-6-sulfonihappo):n avulla. Lakkaasin kiinnittymistä membraaniin tutkittiin ATR-FTIR spektroskoopilla kiinnittymisen varmentamiseksi. Lakkaasi modifioituja membraaneja testattiin koivu-uute suodatuksella ja adsorptio kokeella. Lakkaasientsyymi saatiin kiinnitettyä membraaniin ristisilloittajan avulla, mutta lakkaasilla modifioitujen membraanien vesivuot laskivat noin puoleen alkuperäisestä. Koivu-uuteen suodatuksissa modifioidusta membraanista ei saatu permeaattia lävitse, mutta adsorptiokokeen tulosten perusteella voidaan todeta lakkaasientsyymin pilkkoneen ligniiniä. Kiinnitetyn lakkaasin aktiivisuus vaihteli rinnakkaisten määritysten välillä, minkä vuoksi lakkaasin kiinnitysmekanismin lisätutkiminen olisi tarpeen luotettavimpien tulosten saamiseksi.