O controle bacteriano na indústria alimentícia é de extrema importância uma vez que os microrganismos são responsáveis por causar contaminações persistentes, levando à deterioração do alimento e à transmissão de doenças. Este trabalho teve por objetivo avaliar o potencial antimicrobiano do biossurfatante ramnolipídeo (RL) e de óleoresinas (OR) de aipo, noz moscada, alho, gengibre, pracaxi, patauá e buriti frente as bactérias patogênicas alimentares Listeria monocytogenes, Bacillus cereus e Escherichia coli enterohemorrágica (EHEC). O OR de aipo foi avaliado individualmente e em combinação com o RL. A atividade antimicrobiana foi determinada pelo método de microdiluição em caldo - concentração inibitória mínima (CIM) e concentração bactericida mínima (CBM). Biofilmes de L. monocytogenes e B. cereus foram formados em placas de microtitulação de poliestireno, respectivamente, nos meios de cultivo triptona de soja com extrato de levedura (TSYE) e caldo nutriente (CN) à 37ºC por 48 h e avaliados através da quantificação da biomassa e viabilidade celular após tratamento com os antimicrobianos. O RL apresentou efeito bacteriostático com CIM 125 &#956g/mL frente a L. monocytogenes e para B. cereus observou-se ação bactericida com CBM de 31,25 &#956g/mL. A triagem preliminar mostrou que, dentro da faixa das concentrações testadas, os óleos de buriti, pracaxi, patauá, alho e gengibre não apresentaram CIM frente aos patógenos. O OR de aipo inibiu o crescimento de L. monocytogenes e B. cereus com CIM de 3% e 0,75%, respectivamente. A combinação de 125 &#956g/mL de RL e 3% de óleo de aipo foi bacteriostática para L. monocytogenes, enquanto para B. cereus a combinação dos agentes foi bacteriostática com menor valor de CIM (7,81 &#956g/mL de RL e 0,19% de OR de aipo), sugerindo efeito sinérgico. A linhagem de Escherichia coli (EHEC) foi resistente aos agentes nas concentrações testadas. O RL removeu 70,7% dos biofilmes de L. monocytogenes após 24 h de tratamento, entretanto a redução da viabilidade celular foi maior após 2 h de contato com RL inibindo 66,6%. O OR de Aipo (6%) foi capaz de reduzir 57,1% da viabilidade celular após 24 h de tratamento, já a combinação do RL com OR de aipo foi eficaz em todos os tempos de tratamentos, com redução média de 49,5% de células viáveis do biofilme de L. monocytogenes. Para o biofilme de B. cereus o melhor tratamento (13h) com RL (31,25 &#956g/mL) removeu 71,5% do biofilme, enquanto que a combinação com o OR de aipo (15,62 &#956g/mL + 0,38%) foi capaz de remover 79,5% da biomassa. O RL (62,50 &#956g/ mL) mostrou inibição média de 85,4% da viabilidade celular de biofilme de B. cereus. O melhor tratamento com OR de aipo (1,50%) reduziu a viabilidade celular de B. cereus em 51,2%, após 2 h; e quando combinado com RL (15,62 &#956g/ mL + 0,38%) em 71,8%. RL e OR de aipo mostraram ação antimicrobiana frente a células planctônicas e sésseis de L. monocytogenes e B. cereus, sugerindo potencial para desenvolvimento de novas formulações naturais visando o controle destes patógenos alimentares. / Bacterial control in the food industry is very important once many microorganisms are responsible for causing persistent contamination, leading to food deterioration and disease transmission. Research and development of natural antimicrobial agents is a trend of the market that has been gaining increasing interest. The aim of this study was to evaluate the antimicrobial potential of the rhamnolipid biosurfactant (RL) and celery oleoresin (OR) against food pathogens Listeria monocytogenes, Bacillus cereus and Escherichia coli enterohemorrhagic (EHEC) both in planktonic and in biofilms forms. Celery OR was evaluated individually and in combination with RL. Antimicrobial activity was determined by the broth microdilution method and expressed as minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC). Biofilms of L. monocytogenes and B. cereus were formed in polystyrene microtiter plates, respectively, in tryptone yeast extract (TSYE) and nutrient broth (NB) media at 37°C for 48 h. Biofilms were evaluated by quantification of biomass and cell viability after treatment with antimicrobials. RL presented bacteriostatic effect showing MIC of 125 &#956g/mL against L. monocytogenes and a bactericidal effect at 31.25 &#956g/mL against B. cereus. Preliminary screening showed that, within the range of tested concentrations, buriti, pracaxi, patauá, garlic and ginger oils did not show minimal inhibitory concentration against pathogens. Celery OR inhibited the growth of L. monocytogenes and B. cereus at MIC of 3% and 0.75%, respectively. The combination of 125 &#956g/mL RL and 3% celery oil was bacteriostatic for L. monocytogenes, while for B. cereus the combination effect was also bacteriostatic at a lower MIC (7.81 &#956g/mL RL and 0.19% celery OR), suggesting synergistic effect. Escherichia coli strain (EHEC) was resistant to all agents at the concentration tested. The biofilms of L. monocytogenes were removed by 70.7% after 24 h of treatment using RL (250 &#956g/mL), however, at shorter time (2 h) were more effective in reducing cell viability, showing an inhibition of 66,6%. Celery OR (6%) was able to reduce 57.1% of cell viability after 24 h of treatment, and the combination of RL and celery OR was effective at all treatment times, with a mean reduction of 49.5% of viable L. monocytogenes biofilm cells. Biofilms of B. cereus treated with RL (31.25 &#956g/ml) for 13 h were removed in 71.5%, while the combination of RL with celery OR (15.62 &#956g/mL + 0.38%) was able to remove 79.5%. The RL (62.50 &#956g/mL) reduced B. cereus biofilm viability by a mean of 85.4%. Treatment with celery OR (1.50%) reduced the cell viability of B. cereus in 51.2% after 2h and when combined with RL (15.62 &#956g/ mL + 0.38%) in 71.8%. The RL and celery OR showed antimicrobial activity against planktonic and sessile cells of L. monocytogenes and B. cereus suggesting potential to development of new natural formulations to control these food pathogens.

Neste trabalho, investigou-se o crescimento e as interações de microorganismos, anaeróbios, em batelada com células planctônicas e em reatores diferenciais com biofilme, sob condições termofílicas (55ºC). Foram utilizadas técnicas clássicas para caracterizar os microorganismos anaeróbios estritos e biologia molecular (FISH), com sondas fluorescentes específicas, para caraterizar e quantificar os microrganismos pertencentes ao domínios Archea e Bacteria. Foram realizadas experimentos com meios de cultivo contendo acetato de sódio e propionato de sódio, na presença de sulfato de sódio. Os resultados do FISH mostraram, para os ensaios em batelada com acetato e sulfato, o predomínio de organismos pertencentes ao Domínio Archea (ARC915+MB1174) representados pelo gênero Methanosarcina sp. e bacilos fluorescentes. No biofilme, para a mesma condição, houve predomínio de células do Domínio Bacteria (EUB338) e relacionadas com a oxidação do acetato. Para a condição de cultivo com lactato e sulfato, as células bacterianas semelhantes a Desulfotomaculum sp. predominaram nos reatores em batelada e diferenciais, justificando a elevada porcentagem de células do Domínio Bacteria. Os resultados do NMP mostraram uma maior concentração de anaeróbios totais na condição lactato e sulfato (\'10 POT.11\' - \'10 POT.12\'). Para as arqueas metanogênicas essa concentração foi eveidenciada para a condição de cultivo com acetato e sulfato (\'10 POT.4\'). Os resultados desse trabalho sugeriram que em reatores em batelada, na condição de cultivo com acetato e sulfato, ocorreu equilíbrio entre metanogênese, oxidação de acetato e sulfatogênese, enquanto no biofilme, foram favorecidos os dois primeiros tipos de metabolismo. A condição lactato mais sulfato favoreceu a sulfetogênese em ambos os reatores. No reator diferencial, alimentado com proprionato e sulfato ocorreu fermentação com crescimento sintrófico entre oxidadoras de proprionato, metanogênicas e redutoras de sulfato hidrogenotróficas. / In this work, were investigated the growth and interactions of anaerobic microorganisms, in batch and differential reactors with planktonic cells and biofilm, respectively, in thermophilic conditions (55°C). Classic and molecular biology methods (FISH) were utilized to characterize strict anaerobes, with specific fluorescence probes, to characterize the Archaea and Bacteria Domain cells. Experiments with sodium acetate, s01ium lactate and sodium propionate plus sodium sulfate were realized. To the acetate and sulfate batch assays the results of FISH showed the predominance of Archaea Domain cells (ARC915+MB1174), represented by genera Methanosarcina sp. and fluorescents rods. Under the same conditions the biofilm results showed predominance of Bacteria Domain cells (EUB338) and those related to the acetate oxidation. In lactate plus sulfate condition the bacteria cells, as Desulfotomaculum sp., predominated in batch and differential reactors, justifying the high percentage of Bacteria Domain cells. The MPN results showed a high concentration of total anaerobes under lactate plus sulfate condition (\'10 POT.11\' - \'10 POT.12\'). For methanogenic arquea this concentration was obtained in the acetate plus sulfate condition (\'10 POT.4\'). These results suggest a process equilibrium may have occurred among methanogenesis, acetate oxidation and sulfidogenesis in batch reactors under acetate plus sulfate conditions. However, methanogenesis and acetate oxidation metabolisms predominated in the biofilm. Sulfidogenesis was easier in both reactors, under lactate plus sulfate conditions. In the differential reactors with propionate plus sulfate, the fermentation was in syntrophic co-culture among propionate oxidizing bacteria, methanogenic and hydrogenotrophic sulfate reducing bacteria.

Os endoscópios são equipamentos aprovados para serem reutilizados, apesar de apresentarem estrutura interna complexa, composta por canais longos com lúmens estreitos, o que permite a aderência de matéria orgânica e microrganismos, favorecendo assim a formação de biofilmes. Estes dificultam o processamento eficaz, representando um desafio no reuso desses equipamentos. A formação do biofilme é inevitável ao longo do processamento dos endoscópios e há grande dificuldade em removê-lo completamente. Em razão disso, pesquisas atuais apontam-no como possível responsável pela transmissão de infecções exógenas e efeitos adversos que se manifestam em pacientes submetidos a endoscopias gastrointestinais. Essa realidade tem trazido à tona a preocupação em erradicá-lo. Assim, este estudo teve como objetivos avaliar a ação da desinfecção de alto nível na remoção de biofilme após limpeza prévia com escovação em corpos amostrais, simulando os canais de endoscópios flexíveis e comparar os produtos comercialmente disponíveis no mercado nacional para a remoção do biofilme dos endoscópios. Trata-se de uma pesquisa experimental, laboratorial e comparativa. Foram utilizados tubos de revestimento interno de politetrafluoretileno (Teflon®) e a Pseudomonas aeruginosa como microrganismo teste, formadora de biofilme. Foi montado um modelo validado para o desenvolvimento do biofilme. Os corpos amostrais contaminados foram submetidos inicialmente ao processo manual de limpeza com detergente enzimático e escovação. Em seguida, os corpos amostrais foram submetidos a cinco métodos de desinfecção de alto nível, quais sejam: o ácido peracético com concentração de 0,09% a 0,15%, o sistema Steris®, o glutaraldeído a 2%, e a água eletrolítica ácida. Foram extraídos três segmentos, de aproximadamente três milímetros representando o início, meio e fim de cada corpo amostral e analisados com o auxílio da microscopia eletrônica de varredura quanto a presença de biofilme. Conclui-se que nenhum método testado removeu 100% dos biofilmes e que essa remoção depende da interação entre o método de limpeza e a desinfecção posterior. Verificou-se que o método que mais removeu fisicamente o biofilme foi o glutaraldeído a 2% em reprocessadora automática, provavelmente justificado pelo double brushing, uma vez que o equipamento tem uma fase de limpeza no início de seu ciclo. O método que se mostrou mais eficaz na remoção de biofilme e outros resíduos constituídos de bactérias isoladas e da matriz de exopolissacarídeos, foi sistema Steris®. O método que se mostrou menos eficaz na remoção do biofilme e outros resíduos foi a água eletrolítica ácida. Considerando que a água é uma fonte de biofilme, sugeri-se utilizar filtros para a água do enxágüe e a rinsagem final dos canais com álcool. A despeito de tecnologias atualmente disponíveis para o processamento de canais dos endoscópios, nenhum dos métodos testados na presente investigação foi completamente eficaz para remover os biofilmes frente ao desafio imposto. Resta, portanto, recomendar que práticas baseadas em protocolos das sociedades sejam rigorosamente seguidas, apesar do tempo requerido para as boas práticas e optar, preferencialmente pelo processo automatizado, a fim de diminuir o erro humano / Endoscope is equipment approved to be reused in spite of its complex internal structure, consisting of long channels with narrowed lumens, allowing adherence of organic material and microorganisms, favoring formation of biofilm, making difficult an effective procedure, which is a challenge in the reuse of this equipment. The biofilm formation during endoscopic procedures is inevitable and it is very difficult to entirely remove it. Consequently, recent researches report it as the probable factor responsible by transmissions of exogenous infections and for the side effects found in patients submitted to gastrointestinal endoscopy procedures. This picture brought the concern to eradicate it. Thus, the objectives of this study were to evaluate the action of high level disinfection to remove biofilm after prior cleaning and brushing in body samples, simulating flexible endoscopic channels and to compare products commercially available in the national market to remove the biofilm from endoscopes. This is an experimental, laboratory and comparative research. Tubes of internal politetrafluorethylen (Teflon) and the Pseudomonas aeruginosa as a test microorganism to form the biofilm. A validated model was designed to develop the biofilm. The contaminated body samples were initially submitted to manual cleaning process with enzymatic cleansing and brushing. Next, these bodies were submitted to five high-level disinfection methods, as follows: peracetic acid at 0.09% and 0.15% of concentration, Steris System, 2% glutaraldehyde and acid electrolytic water. Three segments were removed, measuring approximately three millimeters, representing the beginning, the middle and the end of each body sample. These segments were analyzed by means of screening electronic microscopy in relation to biofilm presence. It was concluded that no tested method removed 100% of biofilm and that this removal depends on the interaction between the cleaning method and later disinfection. It was observed that the most effective procedure to physically remove the biofilm was the 2% glutaraldehyde, in automatic reprocessing method, probably due to double brushing since the equipment had a cleaning phase at the beginning of the cycle. The most effective method to remove the biofilm and other residues of isolated bacteria and of exopolysaccaride matrix was the SterisSystem method. The less effective method to remove biofilm and other residues was the acid electrolic water. Considering that water is the biofilm source, it was suggested to use filters for the water when cleansing and to rinse the channels with alcohol at the end. Regarding new technologies available for processing endoscope channels, none of the tested methods in the present investigation was totally effective to remove biofilm in face of the challenge presented. So, we may suggest that the good practices recommended by the several existing societies be strictly followed, in spite of the time needed for this process. Automated process is suggested as the best option to decrease human error

Este estudo avaliou a eficiência da oleuropeína (OLE) (composto fenólico extraído das folhas de Oliveira) isolada e associada aos sanitizantes comerciais ácido peracético 2% (APA), hipoclorito de sódio 2% (HS), peróxido de hidrogênio 3% (PH), digluconato de clorexidina 2% (DC), cloreto de benzalcônio 1% (CB) e iodofor 2% (IO), para inativação de células em suspensão e biofilmes monoespécie e multiespécie formados em superfícies de aço inoxidável ou microplaca de poliestireno por Listeria monocytogenes (ATCC 7644), Staphylococcus aureus (ATCC 25923) e Escherichia coli (ATCC 25922), todas classificadas como fortes produtores de biofilmes. Os isolados foram semeados em caldo TSB (caldo tripticase soja), incubados (37°C/24h) e corrigidos a ~108células/mL (escala 0,5 McFarland). Para bactérias em suspensão, a resistência a sanitizantes foi determinada pela Concentração Inibitória Mínima (CIM) em tubos e pelo método de Disco Difusão em Ágar (DDA), no qual as bactérias foram plaqueadas em ágar TSA contendo discos de 6mm de papel filtro embebidos nos sanitizantes. Após a incubação, a medição dos halos de inibição foi feita com paquímetro. Para os ensaios de resistência dos biofilmes aos compostos sanitizantes, foram utilizadas microplacas de poliestireno 96 poços, as quais foram preparadas para incubação-fixação dos biofilmes e submetidas à leitura em espectrofotômetro de ELISA (600 nm). Em seguida, as placas foram lavadas com solução salina tamponada (PBS, pH 7.4) e os sanitizantes inseridos por 1 minuto. Após neutralização com tiossulfato de sódio (5 minutos), as placas foram lavadas com PBS e metanol, coradas com cristal violeta 1% e coradas com ácido acético glacial (33%) para nova leitura a 570nm. A eficácia da remoção do biofilme pelos sanitizantes foi comparada pelo índice de formação de biofilme (IFB). As imagens do aço inoxidável após tratamento com sanitizante foram feitas através de Microscopia Eletrônica de Varredura (MEV) e Microscopia Confocal, para visualizar a persistência dos biofilmes. Os valores de CIM (diluição 1:2) mostraram que OLE não teve atividade bactericida. No método DDA, L. monocytogenes, foi resistente à OLE, enquanto E. coli e S. aureus apresentaram resistência intermediária. Os sanitizantes comerciais apresentaram boa atividade bactericida nos ensaios de CIM e DDA, sendo que as associações de OLE aos sanitizantes comerciais aumentaram o efeito germicida. Nos ensaios com biofilmes em monoespécie, somente os sanitizantes comerciais, isolados ou associados com OLE, foram eficazes de reduzir o valor de BFI em microplaca de poliestireno. Em biofilmes multiespécie, OLE apresentou efeito antimicrobiano, sobretudo sobre a associação de L. monocytogenes + E. coli + S. aureus (redução: 91,49%). Nenhum dos compostos avaliados foi capaz de inativar completamente os biofilmes nas superfícies de aço inoxidável, uma vez que células viáveis foram observadas após os tratamentos com os sanitizantes, indicando persistência dos biofilmes. Os resultados indicam que a oleuropeína apresentou potencial para incrementar o efeito bactericida de sanitizantes comerciais para eliminação de biofilmes em superfícies inertes, sendo necessários estudos para compreender os mecanismos de ação dessas combinações. / This study evaluated the efficiency of oleuropein (OLE) (a phenolic compound extracted from Oliveira leaves) alone or in association with commercial sanitizers peracetic acid 2% (PPA), sodium hypochlorite 2% (SH), hydrogen peroxide 3% (HP), chlorhexidine digluconate 2% (CD), benzalkonium chloride 1% (BC) and iodophor 2% (IO), for inactivation of suspended cells and monospecies or multispecies biofilms formed on stainless steel surfaces or microplate polystyrene by Listeria monocytogenes (ATCC 7644), Staphylococcus aureus (ATCC 25923) and Escherichia coli (ATCC 25922), all classified as strong producers of biofilms. The isolates were grown in TSB (trypticase soy broth), incubated (37°C/24 h) and adjusted to ~108 cells/mL (0.5 McFarland scale). For the bacterial suspensions, resistance to sanitizers was determined by Minimum Inhibitory Concentration (MIC) in tubes and by Agar Disk Diffusion (ADD), in which the bacteria were plated on TSA agar containing 6mm disks of filter paper soaked in sanitizers. After incubation, measurement of the inhibition halos was done using a caliper rule. For the sanitizer resistance assays with biofilms, polystyrene 96-well microplates were prepared for incubation-fixing of biofilms and read in an ELISA spectrophotometer (600 nm). After, the plates were washed with phosphate buffered saline (PBS, pH 7.4) and the sanitizers were placed for 1 minute. After neutralization with sodium thiosulfate (5 minutes), the plates were washed with PBS and methanol, stained with 1% crystal violet and stained with glacial acetic acid (33%) for a new reading at 570 nm. The effectiveness of biofilm removal by each sanitizer was compared using a Biofilm Formation Index (IFB). The images from stainless steel coupons after treatments with sanitizers were obtained by Scanning Electron Microscopy (SEM) and Confocal Microscopy, to view the persistence of biofilms. MIC values (dilution 1: 2) showed that OLE had no bactericidal activity. In the ADD assays, L. monocytogenes was resistant to OLE, while E. coli and S. aureus showed intermediate resistance. Commercial sanitizers showed good bactericidal activity in both CIM and DDA assays, and the associations of OLE with commercial sanitizers increased their germicidal effect. In the monospecies biofilm assays, only commercial sanitizers, isolated or associated with OLE, were effective for reducing the BFI values in polystyrene microplates. In multispecies biofilms, OLE had an antimicrobial effect, especially on the association of L. monocytogenes + E. coli and S. aureus (reduction: 91.49%). None of the compounds evaluated was able to completely inactivate the biofilms on the stainless steel surfaces, since viable cells of bacteria were observed after treatment with sanitizers, indicating persistence of biofilms. The results indicate that oleuropein has the potential to enhance the bactericidal effect of commercial sanitizers to eliminate biofilms on inert surfaces. Further studies are needed to understand the mechanisms of action of these combinations.

Biofilm phenotypes were studied in 32 Staphylococcus aureus strains isolated from store-bought meats and 22 from diseased patients in hospitals. Of the meat-associated strains, 21 were methicillin-resistant Staphylococcus aureus (MRSA) and 11 were methicillin-susceptible Staphylococcus aureus (MSSA). The hospital-associated strains included 15 MRSAs and 7 MSSAs. We studied the robustness and composition of the biofilms produced by these strains. We found that on average hospital-associated strains form more robust biofilms than meat associated strains. The model often used to describe S. aureus biofilm composition includes two biofilm types defined by the presence or absence of polysaccharide intercellular adhesin (PIA), PIA-dependent and PIA-independent respectively. In this model, PIA-independent biofilms are structurally reliant on proteins and extracellular DNA (eDNA) and PIA-dependent are structurally reliant on polysaccharides. Enzymatic degradation of the extracellular matrix can reveal which compounds are essential for the structural integrity of the biofilm, and by this model PIA-independent biofilms should be susceptible to both DNase and proteinase K. We found that hospital-associated strains are, on average, more susceptible to degradation by proteinase K. Interestingly, hospital-associated strains are less susceptible to degradation by DNase than meat-associated strains. Finding that proteinase K and DNase susceptibility for these strains are not linked gives evidence to support the idea that S. aureus biofilm composition can vary greatly from strain to strain and that the PIA-dependent and PIA-independent dichotomy of the standard model may be insufficient to describe the variety of S. aureus biofilm composition and may only apply to the extremes of the spectrum. Additionally, we saw no relationship between MRSA or MSSA strains and biofilm robustness, proteinase K degradation, or DNase degradation. Differences in biofilm characteristics between hospital-associated and meat-associated strains reinforce previous findings that these populations are genetically distinct.

Antibiotic resistance is an ever-growing topic of concern within the medical field causing researchers to examine the mechanisms of resistance to develop new antimicrobials. Bacteria’s ability to form biofilms is one mechanism which aids in antimicrobial resistance. Staphylococcus aureus is of special interest as it is one of the most frequent biofilm-forming bacteria found on medical devices causing infections and posing dangerous threats in a clinical setting. A recently developed antimicrobial gel has been shown to have profound effects on treating bacterial infections and wound healing. This research is centered upon examining the antimicrobial effects of this gel on the three different stages of biofilm formation in clinical and laboratory strains of S. aureus. Through a series of experiments examining the effects this gel has on S. aureus at the stages of biofilm attachment, maturation, and dispersion, the gel has shown significant levels of inhibition. These findings indicate that the novel gel disrupts biofilm forming processes of S. aureus, which provides useful information for fighting infections in the medical field. Further research on the uses and effects of this new gel could lead possibility using the antimicrobial compound for a variety of clinical purposes.

No description available.

Small towns that operate wastewater treatment lagoons struggle to meet ammonia limits in cold weather. Here we report the performance of a lagoon, retrofitted with submerged attached-growth reactors (SAGRsTM), to provide insight on ammonia effluent compliance and optimal SAGR sizing as functions of water temperature. The lagoon-SAGR water resource recovery facility (WRRF) removed 95% of incoming ammonia with 94% attributable to the SAGRs. The high treatment capacity of the two primary SAGRs, evidenced by nearly continuous dissolved oxygen saturation and exceedingly high ammonia removals, suggested the two secondary SAGRs were essentially unnecessary and that all four SAGRs should be reduced in size. Furthermore, without the secondary SAGRs, the primary SAGR effluent would have exceeded the permitted ammonia discharge limit only four times in the 2.5 year study. At its current size, the lagoon-SAGR WRRF never exceeded permitted ammonia limits, but size reductions should be used for future retrofits. To further understand cold-weather ammonia removal in the lagoon-SAGR WRRF, we investigated the effect of increased ammonia loading on biomass and the effect of biofilms on microbial abundance. When ammonia loading to the SAGRs was increased in the fall, the lagoon-SAGR WRRF never exceeded its ammonia permit limit, the kinetic coefficients were maintained (0.5-0.8 d-1) and the NH3 removal rates improved (0.25 kg d-1 in baseline loading to 0.45 kg d-1) despite a large temperature decrease (25 °C to < 16 °C). In the biofilm, ammonia-oxidizing archaea abundance was 10 times greater than the ammonia-oxidizing bacteria abundance suggesting the potential importance of ammonia oxidizing archaea in biofilm mediated systems. Additionally, the ammonia and nitrite transforming microbes in the SAGRs had a diverse range of dissolved oxygen affinities and were more abundant in the biofilm in comparison to the wastewater. Anaerobic ammonium-oxidizing bacteria were abundant in the biofilm even though the film constantly interacted with high dissolved oxygen. We found that two components of a successful lagoon-SAGR WRRF were increased biomass in the SAGRs before cold-weather due to elevated ammonia loading and diverse oxygen affinities in the microbes related to ammonia removal.

Non-healing wounds can claim thousands of lives and costs billions of dollars each year, and nurse-led wound clinics are becoming necessary to fill a gap in care for patients with wounds. Even among certified wound nurses using evidence-based clinical protocols, key considerations of care are being missed. Therefore, this project was focused on developing and validating a new biofilm education module for certified or certification eligible wound care nurses. The aim of the module was to boost clinical assessment knowledge and improve patient outcomes. Benners skill acquisition model informed the development of this project. The design of the project also included a panel of expert wound care nurses using a 5-point Likert questionnaire to provide feedback on the biofilm education module, including evaluating the content, context, relevance, and use in the practice setting. Descriptive analysis provided evidence to inform the revision of the education module. Results of the Likert questionnaire ranged in mean score from 4.6 to 5.0, indicating there was strong agreement among the panel members that the education module met the objectives. The completed education module has been presented to the organization leadership for future implementation. This project supports positive social change by improving nurses' preparation to provide early clinical assessment, intervention, and definitive biofilm eradication treatments, ultimately improving patient outcomes.

A COMPARISON OF THE ANTIMICROBIAL EFFICACY OF SILVER DIAMINE FLUORIDE AND SILVER NITRATE: AN IN VITRO STUDY By: Nicholas L Luke, D.D.S. A thesis submitted in partial fulfillment of the requirements for the degree of Master of Science in Dentistry at Virginia Commonwealth University. Virginia Commonwealth University, May 2018 Thesis Advisor: William O. Dahlke Jr., D.M.D. Pediatric Dentistry, Department Chair Purpose: To determine the antimicrobial efficacy of SDF and SN/NaF. Methods: Three bacterial species were combined to create an in vitro biofilm. Treatment was completed with SN, SN/NaF, SDF, SDF½ or untreated (control). Results: The untreated group demonstrated significantly higher growth than all other treatment groups across the study. On the BHI-plates (1-day), there were significant differences between all treatments except SDF and SDF½. On the BHI-plates (3-days), SN/NaF was not significantly different from SDF or SDF½. On the L-MRS-plates (1-day), both SN treatment groups yielded significantly higher growth than the SDF groups. On the L-MRS-plates (3-days), SN yielded significantly higher growth than SN/NaF, SDF, and SDF½. Conclusion: SDF is more effective than SN/NaF, with the exception of BHI-plates (3-days) only and SN/NaF is more effective than SN on primarily S. mutans and L. acidophilus. There is evidence of a possible antimicrobial tolerance of oral bacteria to silver.