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Doctoral Thesis
DOI
https://doi.org/10.11606/T.10.2023.tde-27072023-170502
Document
Author
Full name
Leticia Soares Franco
E-mail
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2023
Supervisor
Committee
Knöbl, Terezinha (President)
Huenuman, Nilton Erbet Lincopan
Maiorino, Fernando Corleto
Piazza, Roxane Maria Fontes
Sá, Lilian Rose Marques de
Title in Portuguese
Avaliação do efeito bactericida de lisozima em estirpes de Salmonella spp.
Keywords in Portuguese
Lisozima
Salmonella Heidelberg
Salmonella Minnesota
Sanidade avícola
Abstract in Portuguese
Salmonellas são bactérias Gram negativas, da família Enterobacteriaceae, considerada um dos principais patógenos entéricos de transmissão alimentar, com grande importância para a saúde pública. Sorovares de Salmonella spp. com fenótipo de resistência múltipla aos antimicrobianos emergiram na avicultura a partir de 2004, justificando a busca por produtos alternativos no controle da infecção nas aves. A lisozima ou 1,4-β-N-acetilmuramidase é uma enzima natural, presente no sistema imune inato dos animais e seres humanos, que possui propriedade bactericida. O mecanismo de ação utilizado é pela via não-enzimática, na presença de atividade de peptídeos catiônicos antimicrobianos e propriedades hidrofóbicas, induzindo a morte celular por alteração na membrana. O objetivo do trabalho foi avaliar o efeito bactericida da lisozima nos sorovares Salmonella Heidelberg e Salmonella Minnesota. Foram isoladas 44 estirpes de Salmonellaspp., em 4 granjas da região sudeste (SP e MG), sendo 28 identificadas como S. Heidelberg e 16 como S. Minnesota. O percentual de isolados com resistência múltipla aos antimicrobianos foi de 97,72%. A Concentração Bactericida Mínima para 90% das estirpes (CBM90) foi ≤ 15 ppm. Ensaios time-kill em tempos pré-estabelecidos demonstraram que a redução de 71,42% das estirpes de Salmonella Heidelberg entre 4 e 8 horas, embora alguns isolados tenham resistido à exposição por mais de 24 horas em concentrações de 1000 e 2000 ppm. Para Salmonella Minnesota, a redução foi de 62,5%. O desafio in vivo com S. Heidelberg revelou redução da frequência de frangos colonizados após o desafio, com 63,66% de frangos positivos aos 21 dias no controle positivo e 26,08% dos frangos colonizados no grupo tratado com lisozima 1000 ppm. Foram detectadas diferenças estatísticas significativas na contagem de colônias no inglúvio e nos cecos de frangos tratados, na comparação com os grupos controles. Esses resultados sugerem que a lisozima possui potencial como substância alternativa no uso de antimicrobianos nas infecções por sorovares resistentes de S. Heidelberg e S. Minnesota.
Title in English
Evaluation of the bactericidal effect of lysozyme on strains of Salmonella spp.
Keywords in English
Lysozyme
Poultry health
Salmonella Heidelberg
Salmonella Minnesota
Abstract in English
Salmonellas are Gram-negative bacteria, from the Enterobacteriaceae family, considered one of the main enteric pathogens of food transmission, with considerable importance for public health. Serovars of Salmonella spp. with multiple antimicrobial resistance phenotypes emerged in poultry from 2004 onwards, justifying the search for alternative products to control bird infections. Lysozyme or 1,4-β-N-acetylmuramidase is a natural enzyme, present in the innate immune system of animals and humans, which has bactericidal properties. The mechanism of action is based on the non- enzymatic route, the presence of antimicrobial cationic peptide activity and hydrophobic properties, inducing cell death by membrane modification. The aim of this work was to evaluate the bactericidal effect of lysozyme on Salmonella Heidelberg and Salmonella Minnesota serovars. Forty-four strains of Salmonella spp. were isolated from 4 farms in the southeast region (SP and MG), 28 of which were identified as S. Heidelberg and 16 as S. Minnesota. The percentage of isolates with multiple antimicrobial resistance was 97.72%. The Minimum Bactericidal Concentration for 90% of the strains (CBM90) was ≤ 15 ppm. Time-kill assays at pre-established times demonstrated a reduction of 71.42% of most strains of Salmonella Heidelberg, between 4 and 8 hours, although some isolates resisted exposure for more than 24 hours at concentrations of 1000 and 2000 ppm. Salmonella Minnesota isolates presented a reduction of 62.5%. The in vivo challenge with S. Heidelberg revealed a reduction in the frequency of chickens colonized after the challenge, with 63.66% of chickens positive at 21 days in the positive control and 26.08% of chickens colonized in the group treated with lysozyme 1000 ppm. Significant statistical differences were detected in colony counts in the proventriculus and cecum of treated chickens, in comparison with the control groups. These results suggest that lysozyme has potential as an alternative substance in the use of antimicrobials in infections by resistant serovars of S. Heidelberg and S. Minnesota.
 
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Publishing Date
2023-12-13
 
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