Evaluation of the active concentration of two disinfectants based on glutaraldehyde and benzalkonium chloride and antimicrobial activity in vitro against Salmonella Heidelberg and Salmonella Typhimurium

Biosecurity, cleaning and disinfection of swine and poultry facilities are fundamental for the reduction of pathogenic microorganisms of importance for public and animal health. The objective of this work was to compare the levels of active ingredient described on the label and the real levels detected in high-performance liquid chromatography (HPLC) of two disinfectants., then evaluate the antimicrobial activity since, following the Germicidal Sanitizing Action and Disinfectant Detergent (Official Method AOAC 960.09) in four different dilutions with the presence of 3% organic matter during 15 min of contact, against Salmonella Heidelberg and Salmonella Typhimurium (ST). The product "A" presents active levels of agreement according to the label. The content of quantified assets for product "B" was lower than that recorded on the label. The disinfectant "A" was effective in microbiological evaluation while the disinfectant "B" had microbiocidal activity compromised by the deficit of assets.

Influence of EDTA and its Association with Benzalkonium Chloride on Enterococcus faecalis Adhesion to Dentin / Influencia del EDTA y su Asociación con Cloruro de Benzalconio en la Adhesión de Enterococcus faecalis a la Dentina

ABSTRACT: The aim of this in vitro study was to investigate the influence of ethylenediaminetetraacetic acid (EDTA) associated with the benzalkonium chloride (BAK) on the adhesion and formation of Enterococcus faecalis biofilms attached to coated dentin. Discs standard bovine dentin blocks were treated with the coating materials evaluated: Saline solution (control), 17 % EDTA, 17 % EDTA associated with 1 % BAK for 5 minutes and subsequently washed with saline solution. Afterwards, biofilms of E. faecalis (ATCC 29212) were grown on the surface of coated dentin blocks for time intervals of 1 hour and 7 days (n = 20) and were subsequently washed with phosphate-buffered saline (PBS). Bacterial viability and total biovolume were analyzed by confocal laser scanning microscopy (CLSM) using the Live/Dead technique. Nonparametric Kruskal-Wallis followed by Dunn tests were used to determine statistical differences (a = 5 %). The 17 % EDTA + 1 % BAK group showed significantly lower biovolume and bacterial viability values at the end of 1 hour (p < 0.05). After 7 days of contamination, the 17 % EDTA and 17 % EDTA + 1 % BAK groups showed similar results that differed statistically from those of the control group (p < 0.05). The saline solution group showed higher values. The use of BAK associated with EDTA on dentin blocks surfaces before exposure to contamination was able to interfere in the adhesion of E. faecalis to dentin. Also, dentin treatment by BAK associated with a chelating agent influences the secondary biofilm formation, which could have important effects on the long-term success of root canal treatment.

RESUMEN: El objetivo del estudio consistió en investigar in vitro, la influencia del ácido etilendiamino-tetraacético (EDTA) con cloruro de benzalconio (BAK) en la adhesión y formación de biopelículas de Enterococcus faecalis a la dentina. Discos de dentina bovina fueron tratadas con solución salina (control), 17 % de EDTA, 17% de EDTA asociado con 1 % de BAK durante 5 minutos y lavadas con solución salina. Las biopelículas de E. faecalis (ATCC 29212) se cultivaron sobre los discos de dentina durante intervalos de tiempo de 1 hora y 7 días (n = 20), lavados con solución salina tamponada con fosfato (PBS). La viabilidad bacteriana y el biovolumen total se analizaron mediante microscopía de barrido por láser (CLSM) utilizando la técnica Live / Dead. Se realizó prueba no paramétrica de Kruskal-Wallis, seguida por Dunn con una diferencia estadística (a = 5 %). El grupo de 17 % EDTA + 1 % BAK mostró valores significativamente menores de biovolumen y viabilidad bacteriana al final de 1 hora (p < 0,05). Después de 7 días de contaminación, los grupos de 17 % EDTA y 17 % EDTA + 1 % BAK mostraron resultados similares que diferían estadísticamente del grupo control (p < 0,05). La solución salina mostró valores más altos. La asociación de BAK con EDTA antes de la contaminación interfirió en la adhesión de E. faecalis. Además, el tratamiento de la dentina por BAK asociado con EDTA influye en la formación de biopelículas secundarias, lo que podría tener efectos importantes sobre el éxito a largo plazo del tratamiento del conducto radicular.

Eficácia de agentes químicos na desinfecção de tubetes anestésicos odontológicos / Effectiveness of chemical agents in disinfecting dental anesthetic cartridges

Durante o atendimento odontológico, o paciente pode ser exposto a várias fontes de contaminações, por isso a equipe odontológica deve sempre implementar ações de biossegurança. Materiais não autoclaváveis, como os tubetes anestésicos, necessitam ser desinfetados previamente ao seu uso, pois não são estéreis, podendo transmitir patógenos entre os pacientes. Este estudo objetivou avaliar e comparar a eficácia de três soluções desinfetantes na redução da carga microbiana em tubetes de anestésicos odontológicos. Os tubetes anestésicos (n = 31) foram escolhidos aleato-riamente e submetidos a diferentes métodos e agentes desinfetantes (Álcool 70%, Dióxido de Cloro 7%; Cloreto de benzalcônio 5,2% com Polihexametileno biguanida 3,5%). Após a desinfecção por métodos de imersão ou fricção, os tubetes foram semeados em meio de cultura contendo caldo tripticase de soja e incubados (48h/37 ºC). Amostras do meio de cultura líquido foram repicadas e semeadas em ágar tripticase de soja, incubado durante 48h a 37 ºC. O crescimento microbiano foi observado pela presença de unidades formadoras de colônias (UFCs) crescidas no ágar. O estudo concluiu que os produtos Álcool 70% e Cloreto de benzalcônio 5,2% com Polihexametileno biguanida 3,5% demostraram ser mais eficazes na eliminação da carga microbiana dos tubetes pelo método de fricção, e que realmente os tubetes anestésicos tem sua superfície externa contaminada. O estudo comprovou ser o método de fricção do agente desinfetante mais eficaz na redução da carga microbiana comparado a imersão. Dos agentes testados, o Dióxido de Cloro 7% não demonstrou um nível de desinfecção satisfatório.

During dental care, the patient may be exposed to various sources of contamination, so the dental team should always implement biosecurity actions. Non-autoclavable materials such as anesthetic cartridges need to be di-sinfected prior to use because they are not sterile and can transmit pathogens between patients. This study aimed to evaluate and compare the effectiveness of three disinfectant solutions to reduce microbial load in dental anesthetic cartridges. Anesthetic cartridges (n = 31) were randomly chosen and submitted to different methods and disinfectants (70% Alcohol, 7% Chlorine Dioxide; 5.2% Benzalkonium Chloride with 3.5% Polyhexamethylene Biguanide). After immersion or friction methods of disinfection, the tubes were seeded in culture medium containing trypticase soy broth and incubated (48h/37 ºC). Samples of liquid culture medium were picked and seeded in trypticase soy agar, incubated for 48h at 37 ºC. Microbial growth was observed by the number of colonies forming units (CFUs) grown on the agar. The study concluded that 70% Alcohol and 5.2% Benzalko-nium Chloride with 3.5% Polyhexamethylene biguanide have been shown to be most effective in eliminating the microbial contamination of the cartridges by the friction method, and that the anesthetic cartridges actually have contamination of their external surface. The study proved that the friction method is most effective in reducing microbial load compared to immersion. Of the agents tested, 7% Chlorine Dioxide did not show a satisfactory level of disinfection.

Effects of benzalkonium chloride and cyclosporine applied topically to rabbit conjunctiva: a histomorphometric study / Efeitos do uso tópico do conservante cloreto de benzalcônio e ciclosporina na conjuntiva de coelhos: estudo histomorfométrico

ABSTRACT Purpose: Chronic instillation of benzalkonium chloride, a preservative, has inflammatory effects on the ocular surface. However, addition of the anti-inflammatory agent cyclosporine to a therapeutic protocol may mitigate these effects. This study compared the toxic effects of a 0.1% benzalkonium chloride solution and the possible protective effect of 0.05% cyclosporine when applied topically to the rabbit conjunctiva. Methods: Fifteen age- and weight-matched, female New Zealand white rabbits were categorized into three groups and treated for 30 consecutive days. Group 1, 2, and 3 - benzalkonium chloride received 0.1% every 24 h, 0.05% cyclosporine every 6 h, and both treatments, respectively. In each rabbit, the left eye was subjected to treatment and the right eye was a control. The rabbits were euthanized at after the experiment. Goblet cells and blood vessels were then enumerated in conjunctival tissues stained with periodic acid-Schiff and hematoxylin-eosin, respectively. Differences between treated and untreated eyes and between groups were compared using the t-test and analysis of variance. Results: Benzalkonium chloride treatment, with and without cyclosporine, significantly reduced (p≤0.05) in the number of goblet cells in treatment eyes compared with that in respective control eyes. Alternatively, adding cyclosporine to benzalkonium chloride did not prevent the loss of conjunctival goblet cells, and a significant reduction in the number of goblet cells was noted. Benzalkonium chloride-induced significant increase in the number of new blood vessels was mitigated significantly by the addition of cyclosporine. Conclusion: This study demonstrated the magnitude of conjunctival injury caused by chronic instillation of benzalkonium chloride. Although cyclosporine did not mitigate the effects on goblet cells, its addition minimized inflammatory angiogenesis induced by benzalkonium chloride.

RESUMO Objetivo: A instilação crônica de cloreto de benzal­cônio, um conservante, tem efeitos inflamatórios na superfície ocular. No entanto, a adição do agente anti-inflamatório ciclosporina a um protocolo terapêutico pode atenuar esses efeitos. Este estudo comparou os efeitos tóxicos de uma solução de cloreto de benzalcônio a 0,1% e o possível efeito protetor de ciclosporina a 0,05% quando aplicado topicamente à conjuntiva de coelho. Métodos: Quinze coelhos fêmeas brancos da raça Nova Zelândia, pareados por idade e peso, foram categorizados em três grupos e tratados por 30 dias consecutivos. Os grupos 1, 2 e 3 - receberam cloreto de benzalcônio 0,1% a cada 24h, ciclosporina a 0,005% a cada 6h e ambos os tratamentos, respectivamente. Em cada coelho, o olho esquerdo foi submetido a tratamento e o olho direito foi controle. Os coelhos foram submetidos à eutanásia após o experimento. Células caliciformes e vasos sanguíneos foram então enumerados em tecidos conjuntivais corados com ácido periódico-Schiff e hematoxilina-eosina, respectivamente. As diferenças entre os olhos tratados e não tratados e entre os grupos foram comparadas usando o teste t e análise de variância. Resultados: O tratamento com cloreto de benzalcônio, com e sem ciclosporina, reduziu significativamente (p£0,05) o número de células caliciformes nos olhos tratados em comparação com os olhos controle correspondentes. Alternativamente, a adição de ciclosporina ao cloreto de benzalcônio não impediu a perda de células caliciformes conjuntivais, e foi observada uma redução significativa no número de células caliciformes. O aumento significativo induzido pelo cloreto de benzalcônio no número de novos vasos sanguíneos foi significativamente mitigado pela adição da ciclosporina. Conclusão: Este estudo demonstrou a magnitude da lesão conjuntival resultante da instilação crônica de cloreto de benzalcônio. Embora a ciclosporina não tenha atenuado os efeitos nas células caliciformes, sua adição minimizou a angiogênese inflamatória induzida pelo cloreto de benzalcônio.

A new experimental model of intrinsic denervation in ileum from wistar rats through intramural microinjections of benzalkonium chloride / Um novo modelo experimental de desnervação intrínseca em íleo de ratos wistar através de micro injeções intramurais de cloreto de benzalcônio

Extensive literature is available about the intrinsic denervation of segments of the digestive tube through the application of CB in the serosa of the viscera. However, this technique has some disadvantages like causing peritonitis, flanges and high mortality, limiting its use in humans. The aim of the present study was to evaluate the feasibility of benzalkonium chloride (CB) to induce intrinsic chemical denervation, through applications of CB in the intramural ileum of wistar rats, as well as deepen the knowledge about the evolution of neuronal injury caused in the process. We used 40 rats, divided into two groups (control-GC and benzalkonium-GB) of 20 animals each, divided into four sub-groups according to the time of postoperative assessment of 24, 48 hours, 30 and 90 days. The animals were submitted to intramural microinjections of sterile saline solution 0.9% (GC) or benzalkonium chloride (GB) in ileal portion, and subsequent histopathological analysis and immunohistochemistry for evaluation of neuronal injury. A significant decrease (p<0.05) was found of the neuronal myenteric count over time in groups, GB3, GB4 and GB2. The specific positive immunolabeling for H2AX and Caspase-3 confirmed the results obtained in the histopathological evaluation, denoting the ignition of irreversible cell injury in 24 hours, evolving into neuronal apoptosis in 48 hours after application of the CB 0.3%. Under the conditions in which this work was conducted, it can be concluded that the application of CB 0.3% by means of microinjections intramural in the ileal wall is able to induce intrinsic chemical denervation of the diverticulum of wistar rats and that the main mechanism of neuronal death is induction of apoptosis.(AU)

Existe vasta literatura sobre a desnervação intrínseca de segmentos do tubo digestório através da aplicação de CB na serosa da víscera. Entretanto, essa técnica tem a desvantagem de causar peritonite, formação de bridas e alta mortalidade, não sendo factível para eventuais utilizações em humanos. O objetivo do presente estudo foi avaliar a viabilidade do Cloreto de benzalcônio (CB) induzir desnervação química intrínseca, por meio de aplicações intramurais em íleo de ratos wistar, além de aprofundar o conhecimento sobre a evolução da lesão neuronal causada neste processo. Foram utilizados 40 ratos, distribuídos em dois grupos (controle- GC e benzalcônio- GB) de 20 animais cada, subdivididos em quatro subgrupos de acordo com o tempo de avaliação pós-operatória de 24, 48 horas, 30 e 90 dias. Os animais foram submetidos à microinjeções intramurais de solução salina estéril 0,9% (GC) ou de cloreto de benzalcônio (GB) em porção ileal, e posterior análise histopatológica e imuno-histoquímica, para avaliação da lesão neuronal. Houve diminuição significativa (p<0,05) na contagem neuronal mientérica ao longo do tempo nos grupos GB2, GB3 e GB4. A imunomarcação específica positiva para H2AX e Caspase-3 confirmou os resultados obtidos na avaliação histopatológica, denotando início da lesão celular irreversível em 24 horas, evoluindo para apoptose neuronal em 48 horas após a aplicação do CB 0,3%. Nas condições em que este trabalho foi conduzido, é possível concluir que a aplicação de CB 0,3% por meio de microinjeções intramurais na parede ileal é capaz de induzir desnervação química intrínseca da porção ileal de ratos wistar e que o principal mecanismo de morte neuronal é a indução de apoptose.(AU)

Influência de inibidores de metaloproteinases na degradação da camada híbrida / Influence of metalloproteinase inhibitors on hybrid layer degradation

Objetivo: realizar uma revisão de literatura acerca da eficácia de utilização da clorexidina (CHX) e de outros tipos de inibidores de metaloproteinases (MMPs) na resistência de união da camada híbrida. Métodos: a busca bibliográfica foi realizada na base de dados PubMed, nos meses de novembro e dezembro de 2018. A pesquisa ocorreu em três fases, com os descritores previamente selecionados. Foram incluídas publicações dos últimos 10 anos no formato de pesquisas científicas realizadas in vitro ou in vivo. Após análise, obedecendo aos critérios de inclusão e exclusão, foram incluídos sete estudos na presente revisão. Resultados/Revisão de literatura: na interface adesiva, os estudos mostram que as MMPs são ativadas durante a etapa de ataque ácido realizada nos protocolos de aplicação de sistemas adesivos, podendo ser ativada tanto por procedimentos adesivos com condicionamento ácido prévio como por sistemas adesivos autocondicionantes. Além da CHX, outras substâncias foram pesquisadas e se mostraram eficazes na inibição de MMPs. Considerações finais: por meio da inibição da atividade das MMPs, é possível obter uma maior durabilidade da interface adesiva e uma menor degradação hidrolítica do colágeno presente na camada híbrida. (AU)

Objective: to perform a literature review on the efficacy of chlorhexidine (CHX) and other types of metalloproteinase inhibitors (MMPs) on hybrid layer bond strength. Methods: the bibliographic search was performed in PubMed, in the months of november and december of 2018. The research was carried out in three phases with the previously selected descriptors. Publications have been included in the last 10 years in the form of scientific research conducted in vitro or in vivo. After analysis, following the inclusion and exclusion criteria, 7 studies were included in the present review. Results / Literature review: in the adhesive interface, the studies show that the MMPs are activated during the acid attack stage carried out in the application protocols of adhesive systems, and can be activated either by adhesive procedures with prior acid conditioning or self-etching adhesive systems. In addition to CHX, other substances were investigated and shown to be effective in inhibiting MMPs. Final considerations: through the inhibition of the MMPs activity it is possible to obtain a greater durability of the adhesive interface and lower hydrolytic degradation of the collagen present in the hybrid layer. (AU)

Botulinum toxin as a treatment for short bowel syndrome in rats

Abstract Purpose: The denervation of the intestine with benzalkonium chloride (BAC) reduces mortality and improves weight gain in rats with short bowel syndrome (SBS). Nevertheless, translating these promising findings from bench to bedside is not feasible because BAC promotes peritonitis and irreversible denervation which may be followed by an uncontrolled dilatation of the viscera. The use of botulinum toxin (BT) instead of BAC to achieve the denervation of the remaining small intestine in SBS could be an interesting option because it leads to a mild and transient denervation of the intestine. Methods: Here we evaluated the effects of the ileal denervation with BT in rats with SBS by verifying the body weight variation and intestinal morphological parameters. Four groups with 6 animals each were submitted to enterectomy with an ileal injection of saline (group E) or BT (group EBT). Control groups were submitted to simulated surgery with an ileal injection of BT (group BT) or saline (group C - control). Results: We observed that the treatment of the remaining ileum with BT completely reversed the weight loss associated to extensive small bowel resection. Conclusion: This may provide a new promising approach to the surgical treatment of SBS.

Influence of surfactants addition on the properties of calcium hypochlorite solutions

Abstract Objectives The aim of this study was to evaluate the influence of surfactants 0.2% or 0.1% cetrimide (Cet) or 0.008% benzalkonium chloride (BAK) on 2.5% calcium hypochlorite (Ca(OCl)2), and compare to sodium hypochlorite (NaOCl), regarding the properties of pH, free chlorine content, surface tension, contact angle, pulp dissolution and antimicrobial activity. Material and Methods The pH and free chlorine content were evaluated by digital pHmeter and by titration, respectively. Surface tension was measured by the platinum ring technique with a Du Noüy tensiometer. The solution's contact angle in human dentin surfaces was checked by Drop Shape Analyzer software. Bovine pulps were used for pulp dissolution analysis and the dissolving capacity was expressed by percent weight loss. Antimicrobial activity over Enterococcus faecalis was evaluated by the agar diffusion method. Results Surfactants addition to Ca(OCl)2 and NaOCl did not alter the pH, free chlorine content and pulp dissolution properties. Ca(OCl)2 had the highest surface tension among all tested solutions. When surfactants were added to Ca(OCl)2 and NaOCl, there was a significant reduction of surface tension and contact angle values. The addition of 0.2% or 0.1% Cet enhanced antimicrobial activity of both Ca(OCl)2 and NaOCl. Conclusion Surfactant addition to 2.5% Ca(OCl)2 has shown acceptable outcomes for pH, free chlorine content, surface tension, contact angle, pulp dissolution and antimicrobial activity. Furthermore, the addition of 0.2% Cet showed better results for all tested properties.

Dermatite de contato por cloreto de benzalcônio / Contact dermatitis from benzalkonium chloride

O cloreto de benzalcônio é um conservante encontrado em produtos de higiene pessoal e preparações farmacêuticas há mais de seis décadas. Antes tido como irritante; porém, evidências crescentes apontam que ele possa induzir alergia de contato em uma taxa mais elevada do que o previsto. Os autores apresentam um caso de um adulto de 71 anos, com reação alérgica ao cloreto de benzalcônio contido em solução tópica oftalmológica. Pretende-se, com este caso, alertar sobre as possíveis hipersensibilidades aos conservantes de medicamentos e cosméticos.

Benzalkonium chloride is a preservative found in personal care products and pharmaceutical preparations for over six decades. It was previously thought to be an irritant, but a growing body of evidence suggests that it may induce contact allergy at a higher rate than anticipated. The authors describe the case of a 71-yearold male patient with allergic reaction to benzalkonium chloride present in a topical ophthalmic solution. With this case report we intend to warn about possible hypersensitivity reactions to preservatives contained in medicines and cosmetics.

Ophthalmic adverse effects of nasal decongestants on an experimental rat model / Efeitos oftálmicos adversos de descongestionantes nasais em modelo experimental com ratos

ABSTRACT Purpose: To investigate the potential effects of chronic exposure to a nasal decongestant and its excipients on ocular tissues using an experimental rat model. Methods: Sixty adult male Wistar rats were randomized into six groups. The first two groups were control (serum physiologic) and Otrivine® groups. The remaining four groups received the Otrivine excipients xylometazoline, benzalkonium chloride, sorbitol, and ethylene diamine tetra acetic acid. Medications were applied into both nostrils twice a day for 8 weeks. Before the rats were sacrificed, epithelial staining, the Schirmer test, and intraocular pressure measurements were performed under ketamine/xylasine anesthesia (50 and 5 mg/kg, respectively). Results: Epithelial defects and dry eye were common findings in all study groups. Cataracts developed in two cases clinically. Histopathological evaluation revealed many different pathological alterations in all parts of the ocular tissues such as corneal edema, polypoid proliferation and hyalinization of the vessel wall, cystic formation of the lens, retinal nerve fiber layer degeneration, and corpora amylacea formation of the lacrimal gland. Conclusions: Prolonged usage of the nasal decongestant xylometazoline and its excipients may cause ophthalmic problems such as dry eyes, corneal edema, cataracts, retinal nerve fiber layer, and vascular damage in rats. Although these results were obtained from experimental animals, ophthalmologists should keep in mind the potential ophthalmic adverse effects of this medicine and/or its excipients and exercise caution with drugs containing xylometazoline, ethylene diamine tetra acetic acid, benzalkonium chloride and sorbitol for patients with underlying ocular problems.

RESUMO Objetivo: Investigar os possíveis efeitos da exposição crônica de descongestionante nasal e seus excipientes em tecidos oculares, utilizando um modelo experimental com ratos. Métodos: Sessenta ratos Wistar adultos machos foram divididos aleatoriamente em seis grupos. Os primeiros dois grupos foram controle (soro fisiológico) e Otrivina®. Os quatro grupos restantes receberam os excipientes de Otrivina, tais como Xilometazolina, Benzalcônio, Sorbitol e Ácido Etilenodiamino Tetracético (EDTA). Os medicamentos foram aplicados em ambas as narinas dos ratos, duas vezes ao dia, durante 8 semanas. Antes que os ratos fossem sacrificados, a coloração epitelial, o teste de Schirmer e a medida da pressão intraocular foram realizados sob anestesia com Ketamina/Xilasina (50 e 5 mg/kg, respectivamente). Resultados: Defeitos epiteliais e olho seco foram achados comuns nos grupos de estudo. A catarata desenvolveu-se clinicamente em dois casos. A avaliação histopatológica revelou a existência de alterações em todas as partes dos tecidos oculares, tais como edema de córnea, proliferação polipoide e hialinização da parede vascular, formação cística da lente, degeneração da camada de fibra nervosa da retina (RNFL) e formação de corpos amiláceos da glândula lacrimal. Conclusões: O uso prolongado do descongestionante nasal Xilometazolina e seus excipientes pode causar vários problemas oftalmológicos, como olho seco, edema de córnea, catarata, RNFL e dano vascular em ratos. Embora esses resultados tenham sido obtidos a partir de animais experimentais, os oftalmologistas devem ter em mente os potenciais efeitos oftalmológicos adversos desse medicamento e/ou de seus excipientes.

Modulation of drug resistance and biofilm formation of Staphylococcus aureus isolated from the oral cavity of Tunisian children

Abstract Objectives: This study aims to investigate the antimicrobial and the anti-biofilm activities of Lactobacillus plantarum extract (LPE) against a panel of oral Staphylococcus aureus (n = 9) and S. aureus ATCC 25923. The in vitro ability of LPE to modulate bacterial resistance to tetracycline, benzalchonium chloride, and chlorhexidine were tested also. Methods: The minimum inhibitory concentrations (MICs) and the minimal bactericidal concentrations of Lactobacillus plantarum extract, tetracycline, benzalchonium chloride and clohrhexidine were determined in absence and in presence of a sub-MIC doses of LPE (1/2 MIC). In addition, the LPE potential to inhibit biofilm formation was assessed by microtiter plate and atomic force microscopy assays. Statistical analysis was performed on SPSS v. 17.0 software using Friedman test and Wilcoxon signed ranks test. These tests were used to assess inter-group difference (p < 0.05). Results: Our results revealed that LPE exhibited a significant antimicrobial and anti-biofilm activities against the tested strains. A synergistic effect of LPEs and drug susceptibility was observed with a 2–8-fold reduction. Conclusion: LPE may be considered to have resistance-modifying activity. A more detailed investigation is necessary to determine the active compound responsible for therapeutic and disinfectant modulation.

The antibacterial activity and release of quaternary ammonium compounds in an orthodontic primer

The aim of this study was to evaluate the impact of 10 wt% benzalkonium chloride (TBBAC) or 10 wt% cetylpyridinium chloride (TBCPC) on the antimicrobial properties of the orthodontic adhesive primer, Transbond XT™ (TB). Antimicrobial activity was assessed using a zone of inhibition diffusion test and the release of the antimicrobial compounds was monitored by high performance liquid chromatography (HPLC). Shear bond strength (SBS) was tested using bovine enamel. Control, TB, specimens failed to demonstrate intrinsic antibacterial activity at 1, 7 and 14 days; whereas, TBBAC and TBCPC showed antibacterial effects at all times. HPLC analysis indicated no significant differences in the release behaviour of TBBAC and TBCPC (ttest, p > 0.05), except for the 7day release which was higher for TBBAC (p < 0.05). By 14 days the extents of release were 27 ± 2% and 25 ± 5% of the total initial loading for TBBAC and TBCPC, respectively. The incorporation of 10 wt% BAC or CPC in Transbond XT™ adhesive primer also resulted in superior shear bond strength at 7 and 14 days (Fisher's LSD, p < 0.05) with no significant change in the mode of bracket failure under shear stress (Pearson's chisquared, p > 0.05) (AU)

El objetivo de este estudio fue evaluar el impacto del cloruro de benzalconio al 10% en peso del peso (TBBAC) o de cloruro de cetilpiridinio al 10% del peso (TBCPC) con propiedades antimicrobianas presentes en el adhesivo acondicionador ortodóncico, Transbond XT ™ (TB). La actividad antimi crobiana se evaluó usando una zona de prueba de difusión de inhibición y la liberación de los compuestos antimicrobianos se controló mediante cromatografía líquida de alta resolución (HPLC). La resistencia de adhesión al corte (SBS) se probó usando esmalte bovino. Las muestras control, TB no lograron demostrar actividad antibacteriana intrínseca a 1, 7 y 14 días; mientras que TBBAC y TBCPC mostraron efectos antibac terianos en todo momento. El análisis por HPLC no indicó diferencias significativas en el comportamiento de liberación de TBBAC y TBCPC (prueba t, p> 0,05), excepto en la liberación a los 7 días que fue más alta para TBBAC (p <0,05). A los 14 días, los grados de liberación fueron de 27 ± 2% y de 25 ± 5% de la carga inicial total para TBBAC y TBCPC, respectivamente. La incorpora ción de 10% en peso de BAC o CPC en el imprimador adhesivo Transbond XT ™ también dio como resultado una resistencia superior corte a los 7 y 14 días (Fisher's LSD, p <0.05) sin cambios significativos en el modo de falla del bracket bajo tensión de corte (Pearson's chicuadrado, p> 0.05) (AU)

Effects of prostaglandin analogs on blood flow velocity and resistance in the ophthalmic artery of rabbits / Efeitos dos análogos da prostaglandina na velocidade do fluxo sanguíneo e resistência na artéria oftálmica de coelhos

ABSTRACT Purpose: The aim of this study was to investigate the effects of prostaglandin analogs on blood flow in the ophthalmic artery of clinically healthy rabbits. Methods: Fifty-five clinically healthy New Zealand white rabbits were divided into six groups, and the left eyes were treated for four weeks with the preservative benzalkonium chloride (BAK) only or a topical formulation of different prostaglandin analogs (bimatoprost BAK, tafluprost BAK-free, travoprost BAK, travoprost POLYQUAD, and latanoprost BAK). Color Doppler imaging was performed before and after the treatments. The mean values of the peak systolic velocity (PSV) and end diastolic velocity and the resistive index (RI) were calculated. Statistical analysis was performed to compare the differences pre- and post-treatment for each drug and post-treatment among the drugs. Results: The prostaglandin analogs did not affect PSV. Bimatoprost BAK, travoprost POLYQUAD, and latanoprost BAK did not change RI. Tafluprost BAK-free and travoprost BAK therapy resulted in similar reductions in RI. No significant differences pre- and post-treatment were found when BAK was administered alone. Conclusion: The prostaglandin analogs tafluprost BAK-free and travoprost BAK improved blood flow in the ophthalmic artery in healthy New Zealand white rabbits, which suggests that these drugs enhance the prevention of the progression the progression of glaucoma.

RESUMO Objetivo: O objetivo deste estudo foi investigar os efeitos dos análogos da prostaglandina (PGAs) no fluxo sanguíneo da artéria oftálmica em coelhos. Métodos: Cinquenta e cinco coelhos da raça Nova Zelândia clinicamente saudáveis foram divididos em seis grupos para tratamento com formulação tópica de diferentes APGs (bimatoprosta BAK, tafluprosta BAK-free, travoprosta BAK, travoprosta POLYQUAD e latanoprosta BAK) e formulações contendo apenas o conservante cloreto de benzalcônio (BAK). Foi realizada ultrassonografia com Doppler antes e após os tratamentos. Os valores do pico da velocidade sistólica (PSV) e da velocidade diastólica final foram obtidos e o índice de resistência (RI) foi então calculado. A análise estatística foi realizada para comparar as diferenças entre cada droga no pré e pós-tratamento, além das diferenças no pós-tratamento entre as drogas. Resultados: Estes colírios PGAs não afetaram o PSV. A bimatoprosta com o conservante BAK, travoprosta com o conservante POLYQUAD e latanoprosta com o conservante BAK não alteraram o RI. Já o tratamento com tafluprosta sem conservante (BAK-free) e travoprosta com o conservante BAK promoveram redução similar dos valores do RI. Não houve diferença significativa na comparação entre valores pré e pós-tratamento quando BAK foi administrado isoladamente. Conclusão: Os PGAs tafluprosta BAK-free e travoprosta BAK melhoraram o fluxo sanguíneo na artéria oftálmica em coelhos da raça Nova Zelândia sugerindo que estes medicamentos possam contribuir na prevenção da progressão do glaucoma.

Lacrimal osmolarity and ocular surface in experimental model of dry eye caused by toxicity / Osmolaridade lacrimal e superfície ocular em modelo de olho seco por toxicidade

Purpose: Describe an animal model of dry induced by topical instillation of BAK and evaluate ocular surface biomarkers and histological findings. Methods: Male Wistar rats were used.Topical instillation of 0.2% BAK eyedrops twice a day during 7 days, in the right eye of each animal, while the other eye was taken as control. After 7 days treatment, we performed evaluation of tear film osmolarity, the red phenol thread and ocular surface staining with fluorescein and lissamine green. Afterwards, the animals were sacrificed for tissue extraction and histological evaluation under optical microscopy and H&E staining. Results: Compared with untreated controls, the BAK-group presented tear secretion significantly decreased, increased ocular surface staining by fluorescein and lissamine green and tear film hyperosmolarity (p <0,05). Histological evaluation revealed epithelial thinning and estromal oedema. Conclusions: A toxicity animal model of dry eye induced by topical instillation of benzalkonium chloride, which presents corneal and ocular surface alterations, decreased tear film volume and tear hyperosmolarity as seen in dry eye condition.

Objetivo: Descrever um modelo animal de olho seco induzido pela aplicação tópica de cloreto de benzalcônio (BAC) e avaliar marcadores de integridade da superfície ocular e os achados histológicos. Métodos: Foram utilizados ratos wistar machos adultos. Foi realizada a administração tópica de colírio de BAC 0,2% no olho direito de cada animal duas vezes por dia, durante 7 dias, sendo o olho contralateral tido como controle. Após o tratamento foi realizada a avaliação da osmolaridade do filme lacrimal, o teste de fenol vermelho e a coloração com fluoresceína e lisamina verde. Os animais foram sacrificados e os tecidos extraídos para o estudo histológico da córnea, por microscopia óptica, corada com hematoxilina eosina (H&E). Resultados: Comparados com os controles não tratados o grupo BAC apresentou diminuição significativa na secreção lacrimal, defeitos na integridade epitelial da superfície ocular marcada por corantes vitais, fluoresceína e lisamina verde além do aumento da osmolaridade do filme lacrimal (p < 0,05). À avaliação histológica observou-se diminuição da espessura do epitélio e edema estromal induzidos pela aplicação de BAC. Conclusão: O modelo animal de olho seco por toxicidade induzido pela aplicação tópica de cloreto de benzalcônio apresentou alterações estruturais da córnea e da superfície ocular, diminuição do volume lacrimal e hiperosmolaridade da lágrima características dessa condição.

Efeito antibacteriano de antissépticos bucais sobre bactérias facultativas / Antibacterial effect of oral antiseptics on facultative bacteria

Avaliar o efeito antibacteriano de antissépticos bucais sobre bactérias facultativas por meio de testes de difusão em ágar e teste por exposição direta. Metodologia: Cepas de S. mutans (ATCC 25175), E. faecalis (ATCC 29212) e P. aeruginosa (ATCC 27853) foram inoculadas em 7 mL de brain heart infusion (BHI) e incubadas a 37°C por 24 horas. Para o teste de difusão em ágar, 15 placas de Petri com 20 mL de brain heart infusion agar (BHIA) foram inoculadas com 0,1 mL das suspensões microbianas, com auxílio de swabs esterilizados, de modo a se obter um crescimento confluente e uma placa de Petri não foi inoculada. Trinta e seis discos de papel com 9 mm de diâmetro foram imersos nas soluções experimentais de cloreto de cetilpiridínio 0,07%, cloreto de cetilpiridínio 0,075%, gluconato de clorexidina 0,12% e cloreto de benzalcônio 0,13% durante 1 minuto. A seguir, três discos de papel contendo uma das soluções irrigantes foram colocados sobre a superfície do BHIA. As placas foram mantidas por 1 hora em temperatura ambiente e incubadas a 37°C por 48 horas. Os diâmetros dos halos de inibição microbiana foram medidos valendo-se de duas medidas de forma perpendicular entre si, sendo obtida a média de seus comprimentos. Para o teste de exposição direta, 216 cones de papel absorventes esterilizados nº 50 foram imersos na suspensão de micro-organismos por 5 minutos, e a seguir foram colocados em placas de Petri e cobertos com 10 mL de uma das soluções testes. Em intervalos de 1, 5, 10 e 30 minutos, 3 cones de papel absorventes foram retirados do contato com as substâncias, transportados individualmente e imersos em 7 mL de Letheen Broth, e incubados a 37°C por 48 horas. O crescimento microbiano foi avaliado pela turbidade do meio de cultura. Um inóculo de 0,1 mL obtido do Letheen Broth foi transferido para 7 mL de BHI, e incubado nas mesmas condições descritas. O crescimento microbiano foi novamente avaliado pela turbidade do meio de cultura...

Purpose: To evaluate the antibacterial effect of four oral antiseptics (two solutions of cetylpyridinium chloride, chlorhexidine gluconate and benzalkonium chloride) on facultative bacteria using two methods. Methods: Strains were inoculated in 7 mL of brain heart infusion (BHI) and incubated at 37°C for 24 hours. For the agar diffusion test, 15 Petri plates with 20 mL of brain heart infusion agar (BHIA) were inoculated with 0.1 mL of microbial suspensions using sterile swabs to produce confluent growth; one Petri plate was not inoculated. Thirty-six 9-mm paper discs were immersed in the experimental solutions (0.07% cetylpyridinium chloride, 0.075% cetylpyridinium chloride, 0.12% chlorhexidine gluconate, and 0.13% benzalkonium chloride) for 1 minute. Subsequently, three paper discs containing irrigant solutions were placed on the BHIA in each plate. The plates were kept at room temperature for 1 hour and incubated at 37°C for 48 hours. Two measurements of the inhibition zones were made on the paper discs containing the solutions, and mean values were calculated. For the direct exposure test, 216 #50 sterilized paper points were immersed in the microorganism suspensions for 5 minutes, placed onto Petri plates and covered with 10 mL of irrigant solution. At one, five, 10 and 30 minutes, three paper points were removed from the contact substances, transported individually, immersed in 7 mL Letheen broth and incubated at 37°C for 48 hours. Bacterial growth was evaluated by turbidity. An inoculum of 0.1 mL Letheen broth was transferred to 7 mL BHI, and incubated as described above. Bacterial growth was evaluated according to turbidity. Results: Inhibition zones were greater than 10 mm for all substances and all microorganisms under study. The antibacterial...

The prevalence of ocular surface complaints in Brazilian patients with glaucoma or ocular hypertension / Prevalência de sintomas da doença da superfície ocular em pacientes brasileiros com glaucoma ou hipertensão ocular

PURPOSE: To examine the prevalence of ocular surface complaints in Brazilian patients with glaucoma or ocular hypertension who used topical intraocular pressure (IOP)-lowering regimens. METHODS: In this multicenter, noninterventional, single-visit study, adults with glaucoma or ocular hypertension treated with an IOP-lowering regimen were administered the 12-item ocular surface disease index (OSDI) questionnaire. Each response was scored on a 5-point scale, with 0 indicating symptom present none of the time and 4 indicating symptom present all of the time. The average of the 12 item responses for each patient was transformed to a scale from 0 to 100, with higher scores representing worse disabilities. OSDI results then were categorized as absence of OSD (scores of 0-12), mild OSD (scores of 13-22), moderate OSD (scores of 23-32), or severe OSD (scores of 33100). RESULTS: The 173 enrolled patients had a mean age of 61.2 years, were women in 65.3% of cases, and had glaucoma in 89.0% of cases and ocular hypertension in 11.0% of cases. OSDI scores for 158 patients using 1 IOP-lowering therapy indicated no OSD in 37.3% of patients (59/158), mild OSD in 20.9% (33/158), moderate OSD in 17.1% (27/158), and severe OSD in 24.7% (39/158). For the 120 patients using 1 IOP-lowering medication and having a known duration of diagnosis of glaucoma or ocular hypertension, mean OSDI scores were numerically higher (worse) for the 39 patients with a diagnosis ≥6 years long (score 25 [± 20], indicating moderate OSD) than for the 81 patients with a diagnosis lasting <6 years (score 22 [± 20], indicating mild OSD); however, no significant differences in OSDI scores by duration of diagnosis were evident in means (P=0.49), distributions (P≥0.26), or correlation (P=0.77). CONCLUSIONS: A large proportion of Brazilian patients treated with 1 IOP-lowering therapy had some ocular surface complaints.

OBJETIVO: Avaliar a prevalência de sintomas decorrentes de doença de superfície ocular (DSO) em pacientes brasileiros com glaucoma ou hipertensão ocular que utilizam tratamento ocular tópico para redução da pressão intraocular (PIO). MÉTODO: Neste estudo multicêntrico, não intervencional de uma única visita, pacientes adultos com glaucoma ou hipertensão ocular em tratamento para redução da pressão intraocular (PIO) responderam aos 12 itens do questionário "índice de doença da superfície ocular" (OSDI). Cada resposta foi pontuada numa escala de 5 pontos, com 0 (zero) indicando a ausência de sintomas e 4 indicando sintomas presentes todo o tempo. A média de respostas dos 12 itens para cada paciente foi transformada numa escala de 0 a 100, com pontuações mais elevadas representando piores deficiências. Os resultados do OSDI foram categorizados como ausência de DSO (pontuação de 0-12), DSO leve (pontuação de 13-22), DSO moderada (pontuação de 23-32) ou DSO grave (pontuação de 33-100). RESULTADOS: Os 173 pacientes incluídos apresentavam idade média de 61,2 anos, 65,3% eram mulheres (65,3%), tinham glaucoma em 89,0% dos casos e hipertensão ocular em 11,0% dos casos. As pontuações do OSDI para os 158 pacientes utilizando uma medicação para redução da PIO indicaram "DSO ausente" em 37,3% dos pacientes (59/158), "DSO leve" em 20,9% (33/158), "DSO moderada" em 17,1% (27/158) e "DSO grave" em 24,7% (39/158). Para os 120 pacientes utilizando medicação redutora da PIO e com duração conhecida do diagnóstico de glaucoma ou hipertensão ocular, a pontuação média do OSDI foi numericamente superior (pior) para 39 pacientes com diagnóstico realizado há mais de 6 anos (pontuação 25 [± 20] indicando DSO moderado) do que para 81 pacientes com o diagnóstico realizado há menos de 6 anos (pontuação 22 [± 20] indicando DSO leve); no entanto, não houve diferença estatisticamente significativa na média da pontuação OSDI na duração do diagnóstico (P=0.49), distribuição (P≥0,26), ou correlação (P=0,77). CONCLUSÃO: Uma grande proporção de pacientes brasileiros tratados com uma medicação para redução da PIO apresenta sintomas decorrentes de doença da superfície ocular (DSO).

Suscetibilidade a desinfetantes e perfil de resistência a antimicrobianos em isolados de Escherichia coli / Susceptibility to disinfectants and antimicrobial resistance profile in Escherichia coli isolates

A colibacilose, causada por Escherichia coli, é a enfermidade entérica de maior impacto na produção de suínos, podendo levar à morte do animal. Esta bactéria possui grande capacidade de desenvolver resistência a múltiplos antimicrobianos e a desinfetantes. Desta forma, estudos que abordem mecanismos de resistência e perfil de amostras de campo tornam-se necessários. E. coli é amplamente utilizada como modelo de estudos que exploram a resistência intrínseca e extrínseca a multidrogas. Neste trabalho, buscou-se verificar o perfil de sensibilidade de 62 isolados de E. coli de suínos frente a três desinfetantes e a 13 antimicrobianos. Ainda, em 31 destes isolados foi pesquisada a presença de mecanismo de efluxo. Dos três desinfetantes avaliados, o cloreto de alquil dimetil benzil amônio+poliexietilenonilfenileter foi o que se mostrou mais eficaz (100%), seguido do glutaraldeído+cloreto de alquil dimetil benzil amônio (95,2%) e do cloreto de alquil dimetil benzil amônio (88,8%). Dentre os antimicrobianos testados, observou-se maior resistência para a tetraciclina (62,2%) e maior sensibilidade para o florfenicol (88,6%). A alta sensibilidade dos isolados frente aos desinfetantes pode estar relacionada à ausência de mecanismo de efluxo. O índice de resistência múltipla médio aos antimicrobianos foi de 0,52, o que demonstra um perfil multirresistente dos isolados, conduzindo para a necessidade do uso racional destas drogas em suinocultura.

Colibacillosis caused by Escherichia coli is the most important enteric disease in pig production, which may lead to death of the affected animal. The bacterium has a great ability to develop resistance to multiple antibiotics and disinfectants. Thus, investigation addressing mechanisms of resistance and profile of field samples is necessary. E. coli is widely used as a model for studies that explore the intrinsic and extrinsic resistance to multidrugs. In this paper, we attempt to associate the susceptibility profile of 62 isolates of E. coli to three disinfectants and 13 antimicrobials. Also 31 isolates were tested for the presence of efflux mechanism. Of the three disinfectants tested, alkyldimethylbenzylammonium chloride+nonyl phenoxy polyethoxy ethanol was the most effective (100%), followed by glutaraldehyde+alkyldimethylbenzylammonium chloride (95.2%) and alkyldimethylbenzylammonium chloride (88.8%). Among the antimicrobials tested, there was greater resistance to tetracycline (62.2%) and higher sensitivity to florfenicol (88.6%). The high sensitivity of the isolates against disinfectants may be related to the absence of efflux mechanism. The average index of multiple resistance to antimicrobials was 0.52, what demonstrates a profile of multidrug resistant isolates, showing the need for rational use of these drugs in pig production.

Effect of antibacterial agents on the surface hardness of a conventional glass-ionomer cement

In atraumatic restorative treatment (ART), caries removal with hand excavation instruments is not as efficient as that with rotary burs in eliminating bacteria under the glass ionomer cements (GICs). Thus, different antibacterial agents have been used in recent studies to enhance the antibacterial properties of the GICs, without jeopardizing their basic physical properties. OBJECTIVE: The objective of this study was to evaluate the effect of antibacterial agents on the surface hardness of a conventional GIC (Fuji IX) using Vickers microhardness [Vickers hardness number (VHN)] test. MATERIAL AND METHODS: Cetrimide (CT), cetylpyridinium chloride (CPC) and chlorhexidine (CHX) were added to the powder and benzalkonium chloride (BC) was added to the liquid of Fuji IX in concentrations of 1 percent and 2 percent, and served as the experimental groups. A control group containing no additive was also prepared. After the completion of setting reaction, VHN measurements were recorded at 1, 7, 15, 30, 60, and 90 days after storage in 37°C distilled water. A one-way ANOVA was performed followed by a Dunnett t test and Tamhane T2 tests and also repeated measurements ANOVA was used for multiple comparisons in 95 percent confidence interval. RESULTS: VHN results showed significant differences between the control and the experimental groups at all time periods (p<0.05 for all). Significant differences were observed between all study periods for individual groups (p<0.05). After 7 days, VHNs were decreased in all experimental groups while they continued to increase in the control group. BC and CHX groups demonstrated the least whereas CT and CPC groups exhibited most adverse effect on the hardness of set cements. CONCLUSIONS: Despite the decreased microhardness values in all experimental groups compared to the controls after 7 up to 90 days, incorporating certain antibacterial agents into Fuji IX GIC showed tolerable microhardness alterations within the limitations of this in vitro study.

Cloreto de benzalcônio como medicação tópica / Benzalkonium chloride as topical medication

Fontes dos dados: Revisão bibliográfica obtida pela busca nas bases de dados Cochrane, MEDLINE, BIREME, LILACS e SciELO nos últimos 20 anos, utilizando-se os termos: compostos de benzalcônio, mucosa nasal, toxicidade de drogas e rinite. Síntese dos dados: O cloreto de benzalcônio é um composto amônio quaternário que possui ação umectante e detergente, com propriedades emulsificadora e germicida. Os efeitos adversos sobre o organismo têm sido estudados há mais de duas décadas. Ratos tratados com cloreto de benzalcônio por mais de quatro semanas desenvolveram, a partir do sexto dia, hiperemia da região nasal e respiração ruidosa. Em biópsias de tecido nasal de indivíduos saudáveis submetidos ao cloreto de benzalcônio, verificou-se ação deletéria sobre os cílios da mucosa nasal. Outras alterações também foram observadas, como perda da continuidade celular nas amostras de epitélio nasal, defeitos na membrana nuclear, acúmulo de grânulos e perda de organelas. Conclusões: Diferentes efeitos deletérios do cloreto de benzalcônio sobre a mucosa nasal foram demonstrados in vivo e in vitro...

Data sources: Literature review obtained by Cochrane, MEDLINE, BIREME, LILACS and SciELO databases over the last 20 years, using the terms: benzalkonium chloride compounds, nasal mucosa, drug toxicity, and rhinitis. Data synthesis: Benzalkonium chloride is a quaternary ammonium compound that has humectant and detergent actions, and emulsification and germicide properties. The adverse effects on the body have been studied for more than two decades. Mice treated with benzalkonium chloride by more than four weeks, from the sixth day, showed nasal hyperemia and a noisy breathing. In nasal tissue biopsies of healthy individuals treated with benzalkonium chloride a deleterious action on the nasal mucosa cilia was demonstrated. Also, other alterations have been observed: cell loss of continuity in nasal epithelium, nuclear membrane defects, granule accumulation and organelles loss. Conclusions: Different deleterious effects of benzalkonium chloride on the nasal mucosa have been demonstrated in vivo and in vitro. The toxicity was observed even in concentrations below found in commercial preparations and in use for short period of time. Benzalkonium chloride can cause morphological changes in mucous cilia transport; and in immune function possibly clinical implications that may bring injury in the patients...

Intrinsic denervation of the colon is associated with a decrease of some colonic preneoplastic markers in rats treated with a chemical carcinogen

Denervation of the colon is protective against the colon cancer; however, the mechanisms involved are unknown. We tested the hypothesis that the denervated colonic mucosa could be less responsive to the action of the chemical carcinogen dimethylhydrazine (DMH). Three groups of 32 male Wistar rats were treated as follows: group 1 (G1) had the colon denervated with 0.3 mL 1.5 mM benzyldimethyltetradecylammonium (benzalkonium chloride, BAC); G2 received a single ip injection of 125 mg/kg DMH; G3 was treated with BAC + the same dose and route of DMH. A control group (Sham, N = 32) did not receive any treatment. Each group was subdivided into four groups according to the sacrifice time (1, 2, 6, and 12 weeks after DMH). Crypt fission index, ß-catenin accumulated crypts, aberrant crypt foci, and cell proliferation were evaluated and analyzed by ANOVA and the Student t-test. G3 animals presented a small number of aberrant crypt foci and low crypt fission index compared to G2 animals after 2 and 12 weeks, respectively. From the second week on, the index of ß-catenin crypt in G3 animals increased slower than in G2 animals. From the 12th week on, G2 animals presented a significant increase in cell proliferation when compared to the other groups. Colonic denervation plays an anticarcinogenic role from early stages of colon cancer development. This finding can be of importance for the study of the role of the enteric nervous system in the carcinogenic process.