Sequencing and multiple antimicrobial resistance of Pseudomonas fluorescens isolated from Nile tilapia fish in Egypt / Sequenciamento e resistência antimicrobiana múltipla de Pseudomonas fluorescens isoladas de tilápia-do-nilo no Egito

Pseudomonas fluorescens is one of the main causes of septicemic diseases among freshwater fish, causing severe economic losses and decreasing farm efficiency. Thus, this research was aimed to investigate the occurrence of P. fluorescens in Nile Tilapia (O. niloticus) fish in Egypt, gene sequencing of 16SrDNA gene, and antimicrobial susceptibility. P. fluorescens strains were detected in 32% (128/400) of apparently healthy (9%; 36/400) and diseased (23%; 92/400) Nile tilapia fish. The highest prevalence was observed in gills of fish, 31.3% followed by intestine 26.9%, liver 24.2%, and kidneys 17.6%. The PCR results for the 16SrDNA gene of P. fluorescens showed 16SrDNA gene in 30% of examined isolates. Moreover, Homogeny and a strong relationship between strains of P. fluorescens was confirmed using 16SrDNA sequences. Beside the responsibility of 16SrDNA gene on the virulence of P. fluorescens. The results of antimicrobial susceptibility tests revealed that all strains were resistant to piperacillin (100%), followed by ceftazidime (29.7%), and cefepime (25.8%). The strains of P. fluorescence were highly sensitive to cefotaxime (74.2%), followed by ceftriaxone and levofloxacin (70.3% each). Interestingly, 29.7% of strains of P. fluorescens were multiple antimicrobial-resistant (MAR).

Pseudomonas fluorescens é uma das principais causas de doenças septicêmicas em peixes de água doce, causando graves perdas econômicas e diminuindo a eficiência da fazenda. Assim, esta pesquisa teve como objetivo investigar a ocorrência de P. fluorescens em peixes de tilápia-do-nilo (O. niloticus) no Egito, sequenciamento do gene 16S rDNA e suscetibilidade antimicrobiana. Cepas de P. fluorescens foram detectadas em 32% (128/400) de peixes tilápia-do-nilo aparentemente saudáveis ​​(9%; 36/400) e doentes (23%; 92/400). A maior prevalência foi observada nas brânquias dos peixes, 31,3%, seguida pelo intestino 26,9%, fígado 24,2% e rins 17,6%. Os resultados da PCR para o gene 16SrDNA de P. fluorescens mostraram o gene 16SrDNA em 30% dos isolados examinados. Além disso, a homogeneidade e uma forte relação entre cepas de P. fluorescens foi confirmada usando sequências de 16SrDNA. Além da responsabilidade do gene 16SrDNA na virulência de P. fluorescens. Os resultados dos testes de suscetibilidade antimicrobiana revelaram que todas as cepas foram resistentes à piperacilina (100%), seguida pela ceftazidima (29,7%) e cefepima (25,8%). As cepas de P. fluorescens foram altamente sensíveis à cefotaxima (74,2%), seguida pela ceftriaxona e levofloxacina (70,3% cada). Curiosamente, 29,7% das cepas de P. fluorescens eram multirresistentes a antimicrobianos (MAR).

Sequencing and multiple antimicrobial resistance of Pseudomonas fluorescens isolated from Nile tilapia fish in Egypt

Abstract Pseudomonas fluorescens is one of the main causes of septicemic diseases among freshwater fish, causing severe economic losses and decreasing farm efficiency. Thus, this research was aimed to investigate the occurrence of P. fluorescens in Nile Tilapia (O. niloticus) fish in Egypt, gene sequencing of 16SrDNA gene, and antimicrobial susceptibility. P. fluorescens strains were detected in 32% (128/400) of apparently healthy (9%; 36/400) and diseased (23%; 92/400) Nile tilapia fish. The highest prevalence was observed in gills of fish, 31.3% followed by intestine 26.9%, liver 24.2%, and kidneys 17.6%. The PCR results for the 16SrDNA gene of P. fluorescens showed 16SrDNA gene in 30% of examined isolates. Moreover, Homogeny and a strong relationship between strains of P. fluorescens was confirmed using 16SrDNA sequences. Beside the responsibility of 16SrDNA gene on the virulence of P. fluorescens. The results of antimicrobial susceptibility tests revealed that all strains were resistant to piperacillin (100%), followed by ceftazidime (29.7%), and cefepime (25.8%). The strains of P. fluorescence were highly sensitive to cefotaxime (74.2%), followed by ceftriaxone and levofloxacin (70.3% each). Interestingly, 29.7% of strains of P. fluorescens were multiple antimicrobial-resistant (MAR).

Resumo Pseudomonas fluorescens é uma das principais causas de doenças septicêmicas em peixes de água doce, causando graves perdas econômicas e diminuindo a eficiência da fazenda. Assim, esta pesquisa teve como objetivo investigar a ocorrência de P. fluorescens em peixes de tilápia-do-nilo (O. niloticus) no Egito, sequenciamento do gene 16S rDNA e suscetibilidade antimicrobiana. Cepas de P. fluorescens foram detectadas em 32% (128/400) de peixes tilápia-do-nilo aparentemente saudáveis (9%; 36/400) e doentes (23%; 92/400). A maior prevalência foi observada nas brânquias dos peixes, 31,3%, seguida pelo intestino 26,9%, fígado 24,2% e rins 17,6%. Os resultados da PCR para o gene 16SrDNA de P. fluorescens mostraram o gene 16SrDNA em 30% dos isolados examinados. Além disso, a homogeneidade e uma forte relação entre cepas de P. fluorescens foi confirmada usando sequências de 16SrDNA. Além da responsabilidade do gene 16SrDNA na virulência de P. fluorescens. Os resultados dos testes de suscetibilidade antimicrobiana revelaram que todas as cepas foram resistentes à piperacilina (100%), seguida pela ceftazidima (29,7%) e cefepima (25,8%). As cepas de P. fluorescens foram altamente sensíveis à cefotaxima (74,2%), seguida pela ceftriaxona e levofloxacina (70,3% cada). Curiosamente, 29,7% das cepas de P. fluorescens eram multirresistentes a antimicrobianos (MAR).

Characterization of Bacillus cereus isolated from contaminated foods with sequencing of virulence genes in Egypt.

The current study describes the presence of Bacillus cereus (B. cereus) in contaminated foods of animal source and ready for human consumption with highlighting on their virulence contributing factors by detection of its virulence genes in addition to identification of their sequencing. Three hundred sixty food samples categorized as (228) meat products and (132) milk products were examined for B. cereus isolation and all of these isolates were confirmed by biochemical tests. Eighteen strains obtained from different food samples were examined for the attendance of a number of virulence genes (nheA, cytK, entFM, bceT and hblC genes) using uniplex PCR method. Furthermore, the B. cereus strains were valued for the sequencing of described genes. Generally 24.44% (88/360) food samples classified as 11.11% (40/360) meat products and 13.33% (48/360) milk products carried B. cereus according to cultural and biochemical properties, with geometric mean (1.5×107±0.15 CFU/g or mL) . The highest counts (above 105 CFU/g or mL) were originated from milk products (with geometric mean 2.2×107±0.22 CFU/g or mL) more than meat products (with geometric mean 1×107±0.19 CFU/g or mL). The results revealed that all of our isolates had one or more virulence (enterotoxin) genes. In our research, the most predominant genes were nheA (100%), followed by cytK (61.11%), entFM (33.33%), bceT (11.11%) then hblC (5.56%). Molecular method detected that overall, 5 strains (27.78%) harbored only 1 gene (nheA), 7 strains (38.88%) harbored 2 genes which classified as 5 strains (27.78%) (nheA and cytK), 2 strains (11.11%) have (nheA and entFM). Moreover, 5 strains (27.78%) have 3 genes classified as 3 strains (16.67%) harbored (nheA, cytK and entFM), 1 strain (5.56%) had (nheA, cytK and hblC), and 1 strain (5.56%) had (nheA, cytK and bceT). Only 1 strain (5.56%) carried 4 tested virulence genes (nheA, cytK, entFM and bceT) genes. The most prevalent gene in meat and dairy foods was nheA (100%). The nucleotide sequences of (bceT, cytK, entFM, hblC and nheA genes) of B. cereus strains were deposited in GenBank under accession no. (MW911824, MW911825, MW911826, MW911827 and MW911828), respectively. Our study was established to indicate the presence of virulent B. cereus in meat and milk products ready for human consumption as a result of deficient hygienic actions. So, a plain for good hygienic measures should be modified to avoid causing serious health problems to human due to ingestion of such products.

Sequencing and multiple antimicrobial resistance of Pseudomonas fluorescens isolated from Nile tilapia fish in Egypt.

Pseudomonas fluorescens is one of the main causes of septicemic diseases among freshwater fish, causing severe economic losses and decreasing farm efficiency. Thus, this research was aimed to investigate the occurrence of P. fluorescens in Nile Tilapia (O. niloticus) fish in Egypt, gene sequencing of 16SrDNA gene, and antimicrobial susceptibility. P. fluorescens strains were detected in 32% (128\400) of apparently healthy (9%; 36\400) and diseased (23%; 92\400) Nile tilapia fish. The highest prevalence was observed in gills of fish, 31.3% followed by intestine 26.9%, liver 24.2%, and kidneys 17.6%. The PCR results for the 16SrDNA gene of P. fluorescens showed 16SrDNA gene in 30% of examined isolates. Moreover, Homogeny and a strong relationship between strains of P. fluorescens was confirmed using 16SrDNA sequences. Beside the responsibility of 16SrDNA gene on the virulence of P. fluorescens. The results of antimicrobial susceptibility tests revealed that all strains were resistant to piperacillin (100%), followed by ceftazidime (29.7%), and cefepime (25.8%). The strains of P. fluorescence were highly sensitive to cefotaxime (74.2%), followed by ceftriaxone and levofloxacin (70.3% each). Interestingly, 29.7% of strains of P. fluorescens were multiple antimicrobial-resistant (MAR).

Virulence genotyping and antimicrobial resistance profiles of Yersinia enterocolitica isolated from meat and meat products in Egypt.

Pathogenic Yersinia enterocolitica (Y. enterocolitica) is one of the food-borne entero-pathogen responsible for yersiniosis in humans. The purpose of this research was to survey the prevalence, virulence-associated genes, and antimicrobial resistance of Y. enterocolitica isolated from meat and meat product samples in Egypt. Forty-one (5.9%) out of 700- samples of chicken meat, beef, ground beef, and sausage were positive Y. enterocolitica with a high prevalence in chicken meat (12%). Five virulence genes (ail, inv, ystA, ystB, and yadA) were characterized among 41 Y. enterocolitica isolates with variable frequencies. Among the strains tested, the ystB gene was detected with a high percentage (78.1%), followed by inv gene (70.7%), ail gene (14.6%), ystA gene (12.2%), and yadA gene (2.4%). A high resistance rate was estimated to amoxicillin-clavulanic acid (100%), followed by cefazolin (95%), ampicillin (65.9%), and doxycycline (51.2%), whilst a high sensitivity rate was observed to gentamicin and ciprofloxacin (97.6% each). Interestingly, the multidrug resistance was specified in the 70.7% of strains and showing 13 resistance patterns. Based on nucleotide sequence analysis of the 16s rRNA gene, the phylogenetic tree showed the genetic relatedness amongst Y. enterocolitica isolates. These findings highlighted the emergence of virulent and multidrug-resistant pathogenic Y. entrocolitica in retailed meat and meat products in Egypt.

Virulence genotyping and antimicrobial resistance profiles of Yersinia enterocolitica isolated from meat and meat products in Egypt / Genotipagem de virulência e perfis de resistência antimicrobiana de Yersinia enterocolitica isolados de carne e derivados no Egito

Pathogenic Yersinia enterocolitica (Y. enterocolitica) is one of the food-borne entero-pathogen responsible for yersiniosis in humans. The purpose of this research was to survey the prevalence, virulence-associated genes, and antimicrobial resistance of Y. enterocolitica isolated from meat and meat product samples in Egypt. Forty-one (5.9%) out of 700- samples of chicken meat, beef, ground beef, and sausage were positive Y. enterocolitica with a high prevalence in chicken meat (12%). Five virulence genes (ail, inv, ystA, ystB, and yadA) were characterized among 41 Y. enterocolitica isolates with variable frequencies. Among the strains tested, the ystB gene was detected with a high percentage (78.1%), followed by inv gene (70.7%), ail gene (14.6%), ystA gene (12.2%), and yadA gene (2.4%). A high resistance rate was estimated to amoxicillin-clavulanic acid (100%), followed by cefazolin (95%), ampicillin (65.9%), and doxycycline (51.2%), whilst a high sensitivity rate was observed to gentamicin and ciprofloxacin (97.6% each). Interestingly, the multidrug resistance was specified in the 70.7% of strains and showing 13 resistance patterns. Based on nucleotide sequence analysis of the 16s rRNA gene, the phylogenetic tree showed the genetic relatedness amongst Y. enterocolitica isolates. These findings highlighted the emergence of virulent and multidrug-resistant pathogenic Y. entrocolitica in retailed meat and meat products in Egypt.

A Yersinia enterocolitica patogênica (Y. enterocolitica) é um dos enteropatógenos de origem alimentar responsáveis pela yersiniose no ser humano. O objetivo desta pesquisa foi avaliar a prevalência, genes associados à virulência e resistência antimicrobiana de Y. enterocolitica isolada de amostras de carne e produtos à base de carne no Egito. Quarenta e um (5,9%) de 700 amostras de carne de frango, carne bovina, moída e linguiça foram Y. enterocolitica positivas, com alta prevalência em carne de frango (12%). Cinco genes de virulência (ail, inv, ystA, ystB e yadA) foram caracterizados entre 41 isolados de Y. enterocolitica com frequências variáveis. Entre as cepas testadas, o gene ystB foi detectado com uma alta porcentagem (78,1%), seguido pelo gene inv (70,7%), ail genes (14,6%), gene ystA (12,2%) e gene yadA (2,4%). Foi estimada uma alta taxa de resistência ao ácido amoxicilina-clavulânico (100%), seguida de cefazolina (95%), ampicilina (65,9%) e doxiciclina (51,2%), enquanto uma alta taxa de sensibilidade foi observada para gentamicina e ciprofloxacina (97,6% cada). Curiosamente, a resistência a múltiplas drogas foi especificada em 70,7% das cepas e mostrando 13 padrões de resistência. Com base na análise da sequência nucleotídica do gene rRNA 16s, a árvore filogenética mostrou a relação genética entre isolados de Y. enterocolitica. Esses achados destacaram o surgimento de Y. entrocolitica patogênica virulenta e multirresistente em carnes e produtos à base de carne no Egito.

Virulence genotyping and antimicrobial resistance profiles of Yersinia enterocolitica isolated from meat and meat products in Egypt

Abstract Pathogenic Yersinia enterocolitica (Y. enterocolitica) is one of the food-borne entero-pathogen responsible for yersiniosis in humans. The purpose of this research was to survey the prevalence, virulence-associated genes, and antimicrobial resistance of Y. enterocolitica isolated from meat and meat product samples in Egypt. Forty-one (5.9%) out of 700- samples of chicken meat, beef, ground beef, and sausage were positive Y. enterocolitica with a high prevalence in chicken meat (12%). Five virulence genes (ail, inv, ystA, ystB, and yadA) were characterized among 41 Y. enterocolitica isolates with variable frequencies. Among the strains tested, the ystB gene was detected with a high percentage (78.1%), followed by inv gene (70.7%), ail gene (14.6%), ystA gene (12.2%), and yadA gene (2.4%). A high resistance rate was estimated to amoxicillin-clavulanic acid (100%), followed by cefazolin (95%), ampicillin (65.9%), and doxycycline (51.2%), whilst a high sensitivity rate was observed to gentamicin and ciprofloxacin (97.6% each). Interestingly, the multidrug resistance was specified in the 70.7% of strains and showing 13 resistance patterns. Based on nucleotide sequence analysis of the 16s rRNA gene, the phylogenetic tree showed the genetic relatedness amongst Y. enterocolitica isolates. These findings highlighted the emergence of virulent and multidrug-resistant pathogenic Y. entrocolitica in retailed meat and meat products in Egypt.

Resumo A Yersinia enterocolitica patogênica (Y. enterocolitica) é um dos enteropatógenos de origem alimentar responsáveis pela yersiniose no ser humano. O objetivo desta pesquisa foi avaliar a prevalência, genes associados à virulência e resistência antimicrobiana de Y. enterocolitica isolada de amostras de carne e produtos à base de carne no Egito. Quarenta e um (5,9%) de 700 amostras de carne de frango, carne bovina, moída e linguiça foram Y. enterocolitica positivas, com alta prevalência em carne de frango (12%). Cinco genes de virulência (ail, inv, ystA, ystB e yadA) foram caracterizados entre 41 isolados de Y. enterocolitica com frequências variáveis. Entre as cepas testadas, o gene ystB foi detectado com uma alta porcentagem (78,1%), seguido pelo gene inv (70,7%), ail genes (14,6%), gene ystA (12,2%) e gene yadA (2,4%). Foi estimada uma alta taxa de resistência ao ácido amoxicilina-clavulânico (100%), seguida de cefazolina (95%), ampicilina (65,9%) e doxiciclina (51,2%), enquanto uma alta taxa de sensibilidade foi observada para gentamicina e ciprofloxacina (97,6% cada). Curiosamente, a resistência a múltiplas drogas foi especificada em 70,7% das cepas e mostrando 13 padrões de resistência. Com base na análise da sequência nucleotídica do gene rRNA 16s, a árvore filogenética mostrou a relação genética entre isolados de Y. enterocolitica. Esses achados destacaram o surgimento de Y. entrocolitica patogênica virulenta e multirresistente em carnes e produtos à base de carne no Egito.