Aplicação de modelo preditivo do comportamento de Salmonella enterica durante a elaboração de salame de carne suína

O salame de carne suína é um produto pronto para consumo comumente consumido no Brasil no qual a presença de Salmonella enterica tem sido relatada. Durante a sua elaboração, o tratamento térmico não é indicado e por isso a segurança microbiológica depende da qualidade da carne, adição de ingredientes com atividade antimicrobiana, higiene durante a produção, redução de pH e atividade de água (Aw) durante a sua maturação. Além disso, o protocolo de maturação não está determinado na legislação brasileira. Para isso, os modelos matemáticos, que geram previsões do comportamento bacteriano, podem ser utilizados para determinar protocolos de maturação eficientes. Os objetivos deste estudo foram (i) avaliar o comportamento de um coquetel de sorovares de S. enterica (Typhimurium, Infantis e Derby) durante o processo de maturação do salame; (ii) testar o modelo conceito gamma de acordo com Coroller et al. (2015) para a previsão de crescimento / inativação de S. enterica em um estudo Base e (iii) realizar a validação do modelo. Para isso, 50 e 21 salames foram preparados e inoculados com o coquetel de sorovares de S. enterica para o estudo Base e Validação, respectivamente. Os salames foram maturados em diferentes cenários: i. fermentação a 30ºC e secagem a 20ºC no estudo Base; ii. fermentação a 25ºC e secagem a 18ºC no estudo Validação. Amostras periódicas para quantificação de S. enterica, análises de Aw e pH foram realizadas durante a maturação e as curvas foram construídas. Os valores observados no estudo Base foram inseridos no modelo matemático e assim os parâmetros de crescimento / inactivação (μopt, δ1, δ2, α) foram ajustados. Posteriormente, os parâmetros obtidos no estudo Base (μopt = 2,54558, δ1 = 1588,06, δ2 = 163299,72eα = 0,02158) foram aplicados na validação do modelo. As curvas observadas no estudo Base mostraram que a população de S. enterica aumentou 1,23 log UFC.g-1 nas primeiras 21 horas e diminuiu 4,95 log UFC.g-1 após 920 horas de maturação. No estudo Validação, observou-se um aumento de 0,54 log UFC.g-1 nas primeiras 26 horas, seguido de uma inativação de 5,55 log UFC.g-1 após 1095,5 horas de maturação. O valor da Aw representou o limiar na interface de crescimento / inativação; e a inativação bacteriana começou em Aw 0,951. A regressão linear dos valores previstos em comparação aos valores observados mostrou que 97,65% e 95% da variação nos valores observados foram explicados pelos previstos no estudo Base e Validação, respectivamente. Em conclusão, o modelo aplicado mostrou ser capaz de predizer o comportamento de sorovares de S. enterica durante a maturação do salame e, assim, poderá ser aplicado no planejamento de protocolos de fabricação de salame.Salami prepared with pork is a ready-to-eat product commonly consumed in Brazil, and previous studies reported the presence of Salmonella enterica in this product. During the manufacture at industry the heat treatment is usually not performed, therefore food safety will depend on the quality of meat, addition of ingredients with antimicrobial activity, hygiene during production, reduction of pH and water activity. In addition, fermentation and drying duration of salami is not regulated by Brazilian law. Since mathematical models generate predictions that can be used for determining efficient maturation protocols, the aims of this study were to (i) evaluate the behaviour of a S. enterica cocktail of serovars (Typhimurium, Infantis and Derby) during the maturation process of the Brazilian salami; (ii) test the suitability of the "gamma concept" model according to Coroller et al. (2015) for the prediction of growth/inactivation of S. enterica in a Baseline study and (iii) perform the validation of the model. For this, 50 and 21 salami pieces in the Baseline and Validation study, respectively, were prepared and inoculated with the cocktail of S. enterica serovars. Salami pieces were ripened at different scenarios: i. fermentation at 30ºC and drying at 20ºC in the Baseline study; ii. fermentation at 25ºC and drying at 18ºC in the Validation study. Periodical samplings for S. enterica quantification, water activity and pH analyses were performed during maturation and curves were constructed. Observed values in the Baseline study were inserted in the mathematical "gamma concept" model that adjusted growth/inactivation parameters (μopt, δ1, δ2, α). Afterwards the parameters obtained in the Baseline study (μopt = 2,54558, δ1 = 1588,06, δ2 = 163299,72 e α= 0,02158) were applied in the validation of the model. Observed curves in Baseline study showed that S. enterica population increased 1.23 log cfu.g-1 in the first 21 hours and decreased 4.95 log cfu.g-1 after 920 hours of maturation. In the Validation study an increase of 0.54 log cfu.g-1 in the first 26 hours was observed followed by a 5.55 log cfu.g-1 inactivation after 1095,5 hours of maturation. Water activity value represented the threshold in the growth/inactivation interface; and bacterial inactivation started at water activity 0.951. Linear regression of predicted values compared to the observed values showed that 97.65% and 95% of the variation in the observed values could be explained by the predicted ones in the Baseline and Validation study, respectively. In conclusion, the applied model showed to be able to predict the behaviour of Salmonella enterica serovars during the maturation process of salami, and thus the model can be applied in planning of salami manufacture protocols

Isolamento de Salmonella Typhimurium, Listeria monocytogenes e Staphylococcus coagulase positiva de salames comercializados em feiras livres de Porto Alegre, Brasil

Salami is a ready-to-eat (RTE) product frequently purchased at street fairs in Porto Alegre. Salmonella enterica, Listeria monocytogenes, and coagulase-positive Staphylococcus (CPS) are important causes of foodborne disease and can be transmitted through the consumption of RTE foods. The aim of this study was to evaluate the presence of these pathogens in salami sold at street fairs. Ninety salami samples from three commercial brands available at street fairs were analyzed by routine bacteriological methods for Salmonella spp. and Listeria spp., as well as enumeration of CPS. In addition, two samples from each commercial brand were analyzed for water activity (aw). Samples of brand A showed aw values (0.938 and 0.942) above those set by the legislation, while brand B (0.849 and 0.860) and brand C (0.826 and 0.854) were compliant. Microbiological analyses showed that 67.7% were negative to all investigated bacteria. Salmonella Typhimurium was isolated from 4.4% (4/90) of salami samples, all from commercial brand A. Listeria monocytogenes was detected in 3.3% (3/90) of samples, from commercial brands B and C. Moreover, 7.7% (7/90) of samples contained CPS populations non-compliant with legislation. Although the great majority of salami sold at street fairs of Porto Alegre was compliant with standards, S. enterica, L. monocytogenes, and CPS ≥ 5 × 103 cfu.g-1 could be found in this RTE product. Therefore, control measures in the processing industry and consumer’s education about foodborne illness prevention should be maintained.Salame é um alimento pronto para o consumo frequentemente adquirido pela população em feiras livres de Porto Alegre. Salmonella enterica, Listeria monocytogenes e Staphylococcus coagulase positiva são importantes causas de doenças transmitidas por alimentos e podem ser veiculadas por alimentos prontos para o consumo. O objetivo desse estudo foi avaliar a presença desses patógenos em salames vendidos em feiras livres. Noventa amostras de salame pertencentes a três marcas comerciais foram analisados por métodos bacteriológicos de rotina quanto à presença de Salmonella spp. e Listeria spp., bem como enumeração de Staphylococcus coagulase positiva (SCP). Além disso, foi determinada a Atividade de Água (aw) de duas amostras de cada marca comercial. Amostras da marca A apresentaram valores de aw (0,938 e 0,942) acima do permitido na legislação, enquanto as amostras da marca B (0,849 e 0,860) e C (0,826 e 0,854) não violaram esse parâmetro. A análise microbiológica demonstrou que 67,7% das amostras foram negativas para todas as bactérias investigadas. Salmonella Typhimurium foi isolada de 4,4% (4/90) das amostras de salame, todas da marca comercial A. Listeria monocytogenes foi detectada em 3,3% (3/90) das amostras das marcas B e C. Além disso, 7,7% (7/90) das amostras apresentaram SCP acima do número permitido pela legislação. Apesar da grande maioria dos salames comercializados em feiras livres estarem de acordo com a legislação, S. enterica, L. monocytogenes e SCP ≥ 5 × 103 cfu.g-1 podem estar presentes nesse alimento pronto para o consumo. Dessa forma, o controle nas indústrias e a educação dos consumidores sobre a prevenção de doenças transmitidas por alimentos devem ser mantidos

Detection of Genotipically Related Multi-resistant Escherichia coli Isolates in Pig Feces and Carcasses

Background: Antimicrobial resistant bacteria are considered a hazard not only for the treatment of animal diseases but also for public health. Commensal bacteria, such as Escherichia coli are considered a good indicator of antimicrobial resistance in the population, because it is a gut inhabitant and thus undergoes constant pressure of selection by the administration of antimicrobials. Regarding the public health, it is important to evaluate if resistant bacteria carried in the intestinal content of slaughter pigs can be found on the surface of pre chill carcasses. Therefore, the aims of this study were to evaluate the frequency of antimicrobial resistance in E. coli isolated from feces and pig carcasses; and to assess if multi-resistant isolates from both sources were phenotypically and genotypically related.Materials, Methods & Results: Two sampling cycles were conducted in three pig slaughterhouses (A, B and C). In each cycle, samples were collected form: i. feces deposited on the pen floor of the lairage; ii. surface of carcasses at the prechill step. Samples were submitted to a protocol of isolation and confirmation of Escherichia coli. Isolates were grouped according to the origin: feces (n = 355); carcasses (n = 319); and evaluated for antimicrobial resistance by agar diffusion test. Ninety two isolates presenting multidrug resistance profile were analyzed by pulsed-field gel eletrophoresis (PFGE). Among the 674 isolates of E. coli, 7.4% were susceptible to all tested antibiotics while 79.5% (536/674) were multi-resistant. The most frequent resistance patterns were displayed to tetracycline (Tet, 85.9%), ampicillin (Amp, 73.0%), sulfonamide (Sul, 70.0%), florfenicol (Flo, 65.0%) and nalidixic acid (Nal, 58.9%). The most frequent multi-resistance profile among isolates from both origins was [AmpFloNalSulTet]. Multiresistant isolates originated from feces and carcasses displaying genotypically related pulsotypes (≥70% similarity) were found in all three slaughterhouses.Discussion: In agreement with other studies, E. coli isolated from pig feces and carcasses demonstrated a high frequency of antimicrobial resistance and multi-resistance. The most frequent resistance profiles included antimicrobials frequently used on farm as well as drugs that have been banned as feed additives some years ago in Brazil. The selection of resistant strains may be related to the selection pressiondue to the use of antimicrobials in the pig production chain as well as the co-selection of resistance mediated by genes located in common genetic elements. Therefore, the ban of an individual drug is not always associated with the immediate disappearance of the resistance phenotype in the bacteria population. The fact that most multi-resistant E. coli isolates from carcasses belonged to pulsotypes related to those originated from feces samples indicates that resistant E. coli isolates selected on farm may be able to survive the slaughter process and be found on the carcass. In this case, the possibility of those strains being able to reach the population through the consumption of pork products may have to be considered. This hazard has motivated the ban of antimicrobial use in animals in some countries. However, the ban of antimicrobials use on farm is a controversial issue, due to the economical losses that may result from this measure. Therefore, the prudent use of antimicrobials on farm should be encouraged and its influence in the multi-resistance profile of the enteric microbiota should be further studied

Avaliação da sobrevivência de Salmonella enterica inoculada em salame de carne suína

Pork salami is an embedded, cured and ripened product commonly consumed in Brazil, and the presence of Salmonella enterica has already been reported in this product. During its preparation, the microbiological safety depends on the meat quality, addition of ingredients with antimicrobial activity, hygiene during processing, pH and water activity (Aw) reduction during maturation. In Brazil, the maturation protocol has not been determined in food regulation; therefore, the objectives of this study were (a) to identify the fermentation and drying phases during salami maturation; (b) to test the survival of S. enterica during salami processing; and (c) to compare xylose lysine deoxycholate (XLD) and thin agar layer (TAL) agar for recovering Salmonella. The salami samples were prepared with a cocktail of S. enterica strains, fermented at 30°C and dried at 20°C with controlled relative humidity (RH). Periodic sampling for S. enterica quantification and Aw and pH analyses were performed during maturation, and curves were constructed. Fermentation occurred during the first 66 hours, and the pH decreased while the population of S. enterica increased over the first 21 hours. The drying step was able to reduce the bacterial population by approximately 5 log CFU after 875 hours, reaching an Aw of less than 0.78. However, elimination of S. enterica was not achieved. For Salmonella recovery, TAL agar was more efficient than XLD agar.O salame de carne suína é um produto embutido, curado e maturado comumente consumido no Brasil no qual a presença de Salmonella enterica tem sido relatada. Durante a sua elaboração, a segurança microbiológica depende da qualidade da carne, adição de ingredientes com atividade antimicrobiana, higiene durante a produção, redução de pH e atividade de água (Aw) durante a sua maturação. O protocolo de maturação ainda não está determinado na legislação brasileira; portanto o estudo objetivou: (a) identificar as fases de fermentação e dessecação durante a maturação de salame; (b) testar a sobrevivência de S. enterica durante o processamento de salame e (c) comparar os meios de cultura xilose lisina dextrose (XLD) e thin agar layer (TAL) para recuperação de células do referido micro-organismo. Os salames foram elaborados com um coquetel de S. enterica e submetidos à fermentação em 30ºC e secagem a 20ºC com umidade relativa (UR) controlada. Amostragens periódicas para quantificação de S. enterica, análises de Aw e pH foram realizadas durante a maturação e as curvas foram construídas. A fermentação ocorreu nas primeiras 66 horas, quando houve queda do pH do salame; entretanto S. enterica aumentou sua população nas primeiras 21 horas. A etapa de dessecação foi capaz de reduzir aproximadamente 5 log UFC da população bacteriana em 875 horas, alcançando Aw menor que 0,78, mas não foi capaz de eliminar o micro-organismo do alimento. Para enumeração do micro-organismo, o meio sólido TAL foi mais eficiente na recuperação das células submetidas à maturação quando comparado ao ágar XLD comumente utilizado

Detection of Salmonella Heidelberg resistant to colistin in the intestinal content of pigs at slaughter

Salmonella Heidelberg has increasingly been reported as cause of human salmonellosis worldwide. In Brazil, S. Heidelberg has been reported in poultry but it is infrequently isolated from pigs. Here, we describe the isolation of S. Heidelberg resistant to colistin from slaughter pigs. Five pigs and their carcasses belonging to a same slaughter batch in ten consecutive days were sampled for fragment of intestine in the ileocecal region and sponges rubbed on the carcass surface (400 cm2) before chilling. Salmonella detection was performed according to the ISO 6579:2002. Intestinal content was also subjected to Salmonella enumeration by a miniaturized Most Probable Number (MPN) protocol. Salmonella isolates were characterized by antimicrobial resistance by the disk diffusion test, the minimum inhibitory concentration to colistin determination and to gene mcr-1 investigation by PCR. Salmonella was isolated from the intestinal content of 64% (32/50) of the pigs, in amounts that varied from 2.7 to \u3e1,400 MPN/g. Salmonella Heidelberg was the most frequent serovar identified in the intestinal content samples (20/50; 40%), and this serovar was present in eight of the ten pig batches sampled. At the prechill, Salmonella was isolated from 8% of carcasses, and S. Heidelberg was not detected. Salmonella Heidelberg strains were resistant against ampicillin (n=9), tetracycline (n=8), sulfonamide (n=8) and gentamicin (n=5). Nine multi-drug resistant strains were detected; among them four strains were positive for the gene mcr-1. In these strains the MIC value was 8 μg.mL-1, while in the strains without the mcr-1 gene it ranged from 2 μg.mL-1 to 4 μg.mL-1. Therefore, humans in contact with carrier pigs or their environment may be exposed to S. Heidelberg, including strains harboring the gene mcr-1

Escherichia coli in chicken carcasses in southern Brazil : absence of shigatoxigenic (STEC) and isolation of atypical enteropathogenic (aEPEC)

The aim of this study was to investigate the presence of shiga toxin-producing Escherichia coli (STEC) and atypical enteropathogenic Escherichia coli (aEPEC) in frozen chicken carcasses sold at stores in southern Brazil. Typical E. coli colonies were enumerated in 246 chicken carcasses, and the presence of stx1, stx2, eae genes was investigated in their rinse liquid and in E. coli strains isolated from those carcasses. Strains of E. coli were also investigated for the presence of bfp gene. A median of 0.6 cfu.g-1(ranging from <0.1 to 242.7 cfu.g-1) of typical E. coli colonies was found in the carcasses. Shiga toxin-encoding genes (stx1 and stx2) were not detected, indicating that the chicken carcasses were negative for STEC. The intimin protein gene (eae) was detected in E.coli isolated from 4.88% of the carcasses; all tested strains were negative for the bfp gene and were classified as aEPEC. Twenty-two aEPEC strains were tested for resistance to ten antimicrobials and subjected to macrorestriction (PFGE). All the tested aEPEC strains were fully susceptible to cephalosporins, ciprofloxacin and colistin. Resistance to sulfonamide (65%), ampicillin (55%), tetracycline (50%) and gentamicin (45%) were the most frequent. The PFGE profile demonstrated a low level of similarity among the resistant strains, indicating that they were epidemiologically unrelated. The results indicate that aEPEC strains can contaminate chicken meat, and their association with strains implicated in human diarrhea needs to be further investigated

Aplicação de modelo preditivo do comportamento de Salmonella enterica durante a elaboração de salame de carne suína

O salame de carne suína é um produto pronto para consumo comumente consumido no Brasil no qual a presença de Salmonella enterica tem sido relatada. Durante a sua elaboração, o tratamento térmico não é indicado e por isso a segurança microbiológica depende da qualidade da carne, adição de ingredientes com atividade antimicrobiana, higiene durante a produção, redução de pH e atividade de água (Aw) durante a sua maturação. Além disso, o protocolo de maturação não está determinado na legislação brasileira. Para isso, os modelos matemáticos, que geram previsões do comportamento bacteriano, podem ser utilizados para determinar protocolos de maturação eficientes. Os objetivos deste estudo foram (i) avaliar o comportamento de um coquetel de sorovares de S. enterica (Typhimurium, Infantis e Derby) durante o processo de maturação do salame; (ii) testar o modelo conceito gamma de acordo com Coroller et al. (2015) para a previsão de crescimento / inativação de S. enterica em um estudo Base e (iii) realizar a validação do modelo. Para isso, 50 e 21 salames foram preparados e inoculados com o coquetel de sorovares de S. enterica para o estudo Base e Validação, respectivamente. Os salames foram maturados em diferentes cenários: i. fermentação a 30ºC e secagem a 20ºC no estudo Base; ii. fermentação a 25ºC e secagem a 18ºC no estudo Validação. Amostras periódicas para quantificação de S. enterica, análises de Aw e pH foram realizadas durante a maturação e as curvas foram construídas. Os valores observados no estudo Base foram inseridos no modelo matemático e assim os parâmetros de crescimento / inactivação (μopt, δ1, δ2, α) foram ajustados. Posteriormente, os parâmetros obtidos no estudo Base (μopt = 2,54558, δ1 = 1588,06, δ2 = 163299,72eα = 0,02158) foram aplicados na validação do modelo. As curvas observadas no estudo Base mostraram que a população de S. enterica aumentou 1,23 log UFC.g-1 nas primeiras 21 horas e diminuiu 4,95 log UFC.g-1 após 920 horas de maturação. No estudo Validação, observou-se um aumento de 0,54 log UFC.g-1 nas primeiras 26 horas, seguido de uma inativação de 5,55 log UFC.g-1 após 1095,5 horas de maturação. O valor da Aw representou o limiar na interface de crescimento / inativação; e a inativação bacteriana começou em Aw 0,951. A regressão linear dos valores previstos em comparação aos valores observados mostrou que 97,65% e 95% da variação nos valores observados foram explicados pelos previstos no estudo Base e Validação, respectivamente. Em conclusão, o modelo aplicado mostrou ser capaz de predizer o comportamento de sorovares de S. enterica durante a maturação do salame e, assim, poderá ser aplicado no planejamento de protocolos de fabricação de salame.Salami prepared with pork is a ready-to-eat product commonly consumed in Brazil, and previous studies reported the presence of Salmonella enterica in this product. During the manufacture at industry the heat treatment is usually not performed, therefore food safety will depend on the quality of meat, addition of ingredients with antimicrobial activity, hygiene during production, reduction of pH and water activity. In addition, fermentation and drying duration of salami is not regulated by Brazilian law. Since mathematical models generate predictions that can be used for determining efficient maturation protocols, the aims of this study were to (i) evaluate the behaviour of a S. enterica cocktail of serovars (Typhimurium, Infantis and Derby) during the maturation process of the Brazilian salami; (ii) test the suitability of the "gamma concept" model according to Coroller et al. (2015) for the prediction of growth/inactivation of S. enterica in a Baseline study and (iii) perform the validation of the model. For this, 50 and 21 salami pieces in the Baseline and Validation study, respectively, were prepared and inoculated with the cocktail of S. enterica serovars. Salami pieces were ripened at different scenarios: i. fermentation at 30ºC and drying at 20ºC in the Baseline study; ii. fermentation at 25ºC and drying at 18ºC in the Validation study. Periodical samplings for S. enterica quantification, water activity and pH analyses were performed during maturation and curves were constructed. Observed values in the Baseline study were inserted in the mathematical "gamma concept" model that adjusted growth/inactivation parameters (μopt, δ1, δ2, α). Afterwards the parameters obtained in the Baseline study (μopt = 2,54558, δ1 = 1588,06, δ2 = 163299,72 e α= 0,02158) were applied in the validation of the model. Observed curves in Baseline study showed that S. enterica population increased 1.23 log cfu.g-1 in the first 21 hours and decreased 4.95 log cfu.g-1 after 920 hours of maturation. In the Validation study an increase of 0.54 log cfu.g-1 in the first 26 hours was observed followed by a 5.55 log cfu.g-1 inactivation after 1095,5 hours of maturation. Water activity value represented the threshold in the growth/inactivation interface; and bacterial inactivation started at water activity 0.951. Linear regression of predicted values compared to the observed values showed that 97.65% and 95% of the variation in the observed values could be explained by the predicted ones in the Baseline and Validation study, respectively. In conclusion, the applied model showed to be able to predict the behaviour of Salmonella enterica serovars during the maturation process of salami, and thus the model can be applied in planning of salami manufacture protocols

Processo tecnológico e presença de bactérias causadoras de Doença transmitida por alimentos em salames: revisão Bibliográfica

Salame, produto cárneo industrializado curtido, fermentado, maturado, defumado ou não, e dessecado, é um alimento geralmente consumido sem tratamento térmico. As fases do processamento devem garantir a inocuidade do produto a fim de evitar as Doenças Transmissíveis por Alimentos (DTA’s) nos consumidores. Esse produto representa 2% das vendas de toda a aquisição familiar brasileira, sendo consumido por uma grande parcela da população. Visando proteção à saúde dos indivíduos e a qualidade microbiológica dos produtos alimentícios, a Agência Nacional de Vigilância Sanitária (ANVISA) elaborou o Regulamento Técnico sobre Padrões Microbiológicos para Alimentos. Nessa estão determinados os limites e critérios para a tolerância máxima de microrganismos presentes nos alimentos produzidos os quais devem ser rastreados. Nela estão previstos parâmetros para Coliformes a 45ºC, Salmonella sp e Estafilococos coagulase positiva, entretanto este regulamento não obriga o controle de Listeria monocytogenes. Entretanto, a literatura documenta que a presença de Escherichia coli O157: H7, Salmonella Montevideo, estafilococos e Listeria monocytogenes é possível em salame, e muitos surtos foram atribuídos ao consumo do produto. Concluiu-se que esses micro-organismos podem sobreviver no salame, seja por mau processamento, falhas nas Boas Práticas de Fabricação, ou adaptação dos patógenos. Portanto, as autoridades sanitárias devem estar alertas para o monitoramento da inocuidade desse produto.Salami, meat product industrialized tanned, fermented, cured, smoked or not, and dried, is a food consumed without heat treatment. The stages of processing must ensure the safety of the product to prevent foodborne disease to consumers. This product represents 2% of all sales of the Brazilian acquisition family, being consumed by a large proportion of the population. Aiming to protect the health of individuals and the microbiological quality of food products, the Agência Nacional de Vigilância Sanitária (ANVISA) has developed the Technical Regulation on Microbiological Standards for Foods. The limits and criteria for the tolerance of microorganisms present in food produced which should be screened: Coliforms at 45 º C, Salmonella and coagulase-positive Staphylococcus. However this regulation does not require the control of Listeria monocytogenes. The literature documents the presence of Escherichia coli O157: H7, Salmonella Montevideo, staphylococci and Listeria monocytogenes. It was concluded that these microorganisms can survive in this product, whether by bad processing, failures in Good Manufacturing Practices (GMP), either by adaptation of these pathogens. Therefore, health authorities should be alert to monitor the safety of this product