RESUMO
1. Salmonella Gallinarum (SG) infections cause fowl typhoid, which leads to important economic losses. Multidrug resistance (MDR) and the capacity for bacteria to form biofilms could play an important role in the persistence of SG in poultry flocks resulting in intermittent disease outbreaks. The aim of the following study was to assess the lytic activity of two new bacteriophages (Salmonella phages UPF_BP1 and UPF_BP2) against MDR and biofilm-forming SG. 2. Forty-six strains of SG, isolated in 2015, were characterised by antimicrobial resistance, biofilm formation profiles and susceptibility to two new bacteriophages. 3. Of these strains, 24% were multidrug resistant and more than 80% formed biofilm, with no statistical difference between incubation temperatures (42°C or 22°C). With regard to the lytic activity of the phages, 85% of strains were susceptible to at least one phage. Of these, 74% were lysed by both phages, including MDR and biofilm producing strains. 4. The use of salmonella phages UPF_BP1 and UPF_BP2 were shown to be promising alternatives for the biological control of fowl typhoid.
Assuntos
Bacteriófagos , Doenças das Aves Domésticas , Salmonelose Animal , Salmonella enterica , Animais , Biofilmes , Galinhas , Aves DomésticasRESUMO
More than 300 species have been described in the genus Hepatozoon, occurring in different vertebrates. Among these, only Hepatozoon canis and Hepatozoon americanum are seen in dogs. Different methods may be used for laboratory diagnosis. The most common of these is direct parasitological examination of parasite stages in blood smears. The aim of this investigation was to conduct a phylogenetic study on Hepatozoon isolates from symptomatic dogs in the city of Goiânia, Goiás, Brazil. Blood samples were obtained from 40 symptomatic dogs that had been referred to the Veterinary Hospital of the Federal University of Goiás. Among these, only two samples were positive for Hepatozoon spp. using the direct parasitological method. These samples were then subjected to a DNA extraction process and amplification of a fragment of the 18S rRNA by means of PCR. Subsequently, the PCR products from each sample were purified and sequenced. The sequences obtained were then analyzed using the BLASTn algorithm, which identified both sequences of this study as Hepatozoon canis. By applying the Mega4 software, it was confirmed that these isolates of H. canis from dogs in Goiânia are similar to other reference isolates of the same species from other regions of Brazil and worldwide.(AU)
São descritas mais de 300 espécies do gênero Hepatozoon que acometem diferentes vertebrados. Entre estas, apenas Hepatozoon canis e Hepatozoon americanum são descritas em cães. Diferentes métodos podem ser utilizados para o diagnóstico laboratorial. O mais empregado é o exame parasitológico direto do parasito em esfregaços sanguíneos. O objetivo deste trabalho foi realizar um estudo filogenético em Hepatozoon isolados de cães sintomáticos de Goiânia, Goiás. As amostras de sangue foram obtidas de 40 cães sintomáticos encaminhados ao Hospital Veterinário da Universidade Federal de Goiás. Entre essas, duas únicas amostras foram positivas para Hepatozoon spp. pelo método parasitológico direto. Estas amostras foram, então, submetidas ao processo de extração de DNA e de amplificação de um fragmento de 18S rRNA por PCR. Ambas as amostras foram positivas na PCR. Posteriormente, os produtos de PCR de cada amostra foram purificados e sequenciados. As sequências obtidas foram analisadas pelo algoritmo BLASTn, sendo identificadas como Hepatozoon canis. Por meio do software Mega4 foi confirmado que estes isolados de H. canis de cães de Goiânia são semelhantes a outros isolados de referência da mesma espécie de outras regiões do Brasil e do mundo.(AU)
Assuntos
Animais , Cães , Eucoccidiida/isolamento & purificação , Parasitologia , Filogenia , Epidemiologia Molecular , Reação em Cadeia da Polimerase/veterináriaRESUMO
More than 300 species have been described in the genus Hepatozoon, occurring in different vertebrates. Among these, only Hepatozoon canis and Hepatozoon americanum are seen in dogs. Different methods may be used for laboratory diagnosis. The most common of these is direct parasitological examination of parasite stages in blood smears. The aim of this investigation was to conduct a phylogenetic study on Hepatozoon isolates from symptomatic dogs in the city of Goiânia, Goiás, Brazil. Blood samples were obtained from 40 symptomatic dogs that had been referred to the Veterinary Hospital of the Federal University of Goiás. Among these, only two samples were positive for Hepatozoon spp. using the direct parasitological method. These samples were then subjected to a DNA extraction process and amplification of a fragment of the 18S rRNA by means of PCR. Subsequently, the PCR products from each sample were purified and sequenced. The sequences obtained were then analyzed using the BLASTn algorithm, which identified both sequences of this study as Hepatozoon canis. By applying the Mega4 software, it was confirmed that these isolates of H. canis from dogs in Goiânia are similar to other reference isolates of the same species from other regions of Brazil and worldwide.(AU)
São descritas mais de 300 espécies do gênero Hepatozoon que acometem diferentes vertebrados. Entre estas, apenas Hepatozoon canis e Hepatozoon americanum são descritas em cães. Diferentes métodos podem ser utilizados para o diagnóstico laboratorial. O mais empregado é o exame parasitológico direto do parasito em esfregaços sanguíneos. O objetivo deste trabalho foi realizar um estudo filogenético em Hepatozoon isolados de cães sintomáticos de Goiânia, Goiás. As amostras de sangue foram obtidas de 40 cães sintomáticos encaminhados ao Hospital Veterinário da Universidade Federal de Goiás. Entre essas, duas únicas amostras foram positivas para Hepatozoon spp. pelo método parasitológico direto. Estas amostras foram, então, submetidas ao processo de extração de DNA e de amplificação de um fragmento de 18S rRNA por PCR. Ambas as amostras foram positivas na PCR. Posteriormente, os produtos de PCR de cada amostra foram purificados e sequenciados. As sequências obtidas foram analisadas pelo algoritmo BLASTn, sendo identificadas como Hepatozoon canis. Por meio do software Mega4 foi confirmado que estes isolados de H. canis de cães de Goiânia são semelhantes a outros isolados de referência da mesma espécie de outras regiões do Brasil e do mundo.(AU)
Assuntos
Animais , Cães , Eucoccidiida/isolamento & purificação , Parasitologia , Filogenia , Epidemiologia Molecular , Reação em Cadeia da Polimerase/veterináriaRESUMO
Conventional bacteriology techniques and quantitative polymerase-chain reaction (qPCR) were applied to the eggshell, albumen, and yolk of washed and unwashed commercial white and brown eggs, to detect Salmonella spp. Pooled samples of eggshells, albumen, and yolk of white and brown eggs were collected at the poultry house and at the egg-storage room. Salmonella spp. was detected by conventional bacteriology in 5.4% (21/387) of analyzed samples and in 16% (68/387) by qPCR. In the 114 unwashed white eggs samples of eggshell, albumen and yolk, the bacterium was identified in 2.6% of the eggs (3/114) by conventional bacteriology and in 13.2% (15/114) by qPCR. In the 90 samples of washed eggs, 6.7% (6/90) were contaminated as detected by conventional bacteriology and 10.0% (9/90) by qPCR. In the 81 samples of unwashed brown eggs, Salmonella spp. was detected in 6.1% of the eggs (5/81) by conventional bacteriology and 27.2% (22/81) by qPCR. In the 102 samples of brown washed eggs, 6.9% (7/102) where positive by conventional bacteriology and 35.3% (16/102) by qPCR. All samples detected as positive by conventional bacteriology were also positive by qPCR. Salmonella Agona represented 18.2% (4/22) of identified serovars, Salmonella enterica subs. enterica O: 4.5 18.2% (4/22), Salmonella Schwarzengrund 18.2% (4/22), Salmonella Cerro 13.6% (3/22), Salmonella Anatum 13.6% (3/22), Salmonella Enteritidis 9.1% (2/22), Salmonella Johannesburg 4.5% (1/22), and Salmonella Corvallis 4.5% (1/22). The qPCR method provided better detection of Salmonella spp. in commercial eggs than conventional bacteriology. The conventional egg washing and disinfection procedures are not efficient to eliminate Salmonella.
Assuntos
Animais , Casca de Ovo/química , Gema de Ovo/efeitos adversos , Gema de Ovo/química , Reação em Cadeia da Polimerase , Reação em Cadeia da Polimerase/veterinária , SalmonellaRESUMO
Conventional bacteriology techniques and quantitative polymerase-chain reaction (qPCR) were applied to the eggshell, albumen, and yolk of washed and unwashed commercial white and brown eggs, to detect Salmonella spp. Pooled samples of eggshells, albumen, and yolk of white and brown eggs were collected at the poultry house and at the egg-storage room. Salmonella spp. was detected by conventional bacteriology in 5.4% (21/387) of analyzed samples and in 16% (68/387) by qPCR. In the 114 unwashed white eggs samples of eggshell, albumen and yolk, the bacterium was identified in 2.6% of the eggs (3/114) by conventional bacteriology and in 13.2% (15/114) by qPCR. In the 90 samples of washed eggs, 6.7% (6/90) were contaminated as detected by conventional bacteriology and 10.0% (9/90) by qPCR. In the 81 samples of unwashed brown eggs, Salmonella spp. was detected in 6.1% of the eggs (5/81) by conventional bacteriology and 27.2% (22/81) by qPCR. In the 102 samples of brown washed eggs, 6.9% (7/102) where positive by conventional bacteriology and 35.3% (16/102) by qPCR. All samples detected as positive by conventional bacteriology were also positive by qPCR. Salmonella Agona represented 18.2% (4/22) of identified serovars, Salmonella enterica subs. enterica O: 4.5 18.2% (4/22), Salmonella Schwarzengrund 18.2% (4/22), Salmonella Cerro 13.6% (3/22), Salmonella Anatum 13.6% (3/22), Salmonella Enteritidis 9.1% (2/22), Salmonella Johannesburg 4.5% (1/22), and Salmonella Corvallis 4.5% (1/22). The qPCR method provided better detection of Salmonella spp. in commercial eggs than conventional bacteriology. The conventional egg washing and disinfection procedures are not efficient to eliminate Salmonella.(AU)