Galleria mellonella larvae as an alternative model to determine the pathogenicity of avian pathogenic Escherichia coli.

RESEARCH HIGHLIGHTS: Galleria mellonella larvae are a viable model for determining APEC pathogenicity.Larval disease score is the main variable for determining APEC pathogenicity.Response variables should be evaluated up to 24 h post-inoculation.

Development of Predictive Modeling for Removal of Multispecies Biofilms of Salmonella Enteritidis, Escherichia coli, and Campylobacter jejuni from Poultry Slaughterhouse Surfaces.

Salmonella Enteritidis, Escherichia coli, and Campylobacter jejuni are among the most common foodborne pathogens worldwide, and poultry products are strongly associated with foodborne pathogen outbreaks. These pathogens are capable of producing biofilms on several surfaces used in the food processing industry, including polyethylene and stainless steel. However, studies on multi-species biofilms are rare. Therefore, this study aimed to develop predictive mathematical models to simulate the adhesion and removal of multispecies biofilms. All combinations of microorganisms resulted in biofilm formation with differences in bacterial counts. E. coli showed the greatest ability to adhere to both surfaces, followed by S. Enteritidis and C. jejuni. The incubation time and temperature did not influence adhesion. Biofilm removal was effective with citric acid and benzalkonium chloride but not with rhamnolipid. Among the generated models, 46 presented a significant coefficient of determination (R2), with the highest R2 being 0.88. These results provide support for the poultry industry in creating biofilm control and eradication programs to avoid the risk of contamination of poultry meat.

Cutaneous myxoma in broiler chicken detected during slaughtering

Background: A cutaneous or superficial myxoma is a benign neoplasm of dermal or subcutaneous fibroblast origin. Although rare, it has been previously described in several species, including poultry. It presents as a single node or soft mass with a gelatinous cut surface. Histopathological analysis is essential for diagnosis and to differentiate it from other mesenchymal neoplasms and inflammatory or degenerative processes. Microscopically, it consists of dermal or subcutaneous lobules of plump, stellate, or spindle-shaped, bland-looking cells embedded in a basophilic myxoid matrix. This report describes the pathological findings in a rare case of cutaneous myxoma in a 42-day-old broiler flock. Cases: During ante mortem inspection of a 42-day-old broiler flock at a slaughterhouse under the authority of the Federal Inspection Service (southern Brazil), nodular lesions or encrusted areas with yellow and black areas were observed in the head skin of less than 1% of animals. These lesions, approximately 0.5 cm in diameter, were observed on the comb, in the periocular skin region, and close to the animals' nostrils. During the breeding period, no health or epidemiological events were observed. Fragments of the lesions in the comb and periocular skin were collected and fixed in buffered 10% formalin. The samples were sent to the laboratory, routinely processed, and stained with hematoxylin and eosin and Alcian blue. Microscopically, the lesions consisted of irregular multifocal proliferation of connective tissue showing spindle cells with poorly demarcated borders and scarce cytoplasm in a slightly basophilic myxoid aspect matrix. The adjacent epidermis is compressed due to neoplastic proliferation. No areas of epithelial hyperplasia or inclusion bodies were observed. According to the pathologic description and considering its descriptive epidemiology, our main clinical suspicion was cutaneous fowl pox, a pathology characterized by the appearance of nodules in regions devoid of feathers. However, the microscopic changes observed were compatible with those described for cutaneous myxomas. In addition, the extracellular matrix was positive for Alcian Blue staining, which is an indicator of myxoma. In the present case, the SIF did not report the same macroscopic lesions in other flocks of the same origin. Discussion: Connective tissue tumors, including myxomas, occur considerably less frequently under field conditions. In addition, these neoplasms are more frequent in mature birds and are not usually described in broilers, as observed in this report. The cutaneous myxoma described in broilers is usually a sporadic neoplasm that does not cause zootechnical losses, as observed in the case report. Its etiology is unknown and has been associated with various factors, such as local trauma and foreign bodies. Some fragments of plant material from the breeding environment were microscopically detected in the encrusted areas, which may indicate previous trauma or a foreign body. Myxoma has been associated with avian leukosis virus (ALV) subgroup A, but SIF did not report the same macroscopic lesions in other flocks of the same breeder hen's origin in the present case. Furthermore, sporadic connective tissue tumors associated with the virus occur in mature chickens but not in broilers. Myxoma lesions should be considered in the differential diagnosis of other connective tissue tumors and infectious agents that cause lesions in the skin and subcutaneous tissue.

Artificial neural networks for the management of poultry industry: a simulation based on the broiler production chain / Redes neurais artificiais para o gerenciamento da indústria avícola: uma simulação baseada na cadeia de produção de frangos de corte

The aim of this study was to predict production indicators and to determine their potential economic impact on a poultry integration system using artificial neural networks (ANN) models. Forty zootechnical and production parameters from broiler breeder farms, one hatchery, broiler production flocks, and one slaughterhouse were selected as variables. The ANN models were established for four output variables: "saleable hatching", "weight at the end of week 5," "partial condemnation," and "total condemnation" and were analyzed in relation to the coefficient of multiple determination (R2), correlation coefficient (R), mean error (E), mean squared error (MSE), and root mean square error (RMSE). The production scenarios were simulated and the economic impacts were estimated. The ANN models were suitable for simulating production scenarios after validation. For "saleable hatching", incubator and egg storage period are likely to increase the financial gains. For "weight at the end of the week 5" the lineage (A) is important to increase revenues. However, broiler weight at the end of the first week may not have a significant influence. Flock sex (female) may influence the "partial condemnation" rates, while chick weight at first day may not. For "total condemnation", flock sex and type of chick may not influence condemnation rates, but mortality rates and broiler weight may have a significant impact.

O objetivo deste trabalho foi predizer os indicadores de produção e determinar o seu potencial impacto econômico em um sistema de integração utilizando as redes neurais artificiais (RNA). Quarenta parâmetros zootécnicos e de produção de granjas de matrizes e de frango de corte, um incubatório e um abatedouro foram selecionados como variáveis. Os modelos de RNA foram estabelecidos para quatro variáveis de saída ("eclosão vendável", "peso ao final da quinta semana", "condenações parciais" e "condenações totais") e foram analisados em relação ao coeficiente de determinação múltipla (R2), coeficiente de correlação (R), erro médio (E), erro quadrático médio (EQM) e raiz do erro quadrático médio (REQM). Os cenários produtivos foram simulados e os impactos foram estimados. Os modelos de RNA gerados foram adequados para simular diferentes cenários produtivos após o treinamento. Para "eclosão vendável", o modelo de incubadora e o período de incubação aumentaram os ganhos financeiros. Para "peso ao final da quinta semana", a linhagem também demonstrou influencia no retorno financeiro, o que não aconteceu com o peso ao final da primeira semana. O sexo do lote possui influência nas taxas de "condenação parcial", ao contrário do peso do frango no primeiro dia. As taxas de mortalidade e o peso do frango apresentaram influência na "condenação total", mas o sexo do lote e o tipo de pinto não tiverem influência.

Identification of Virulence-Associated Markers in Escherichia Coli Isolated from Captive Red-Browed Amazon Parrot (Amazona Rhodocorytha)

Due to the genetic similarity of pathogenic Escherichia coli isolated from birds and pathotypes of human origin, it is suggested that they have a common ancestor and may exchange virulence-associated genes. This study aimed to detect virulence-associated genes in E. coli strains isolated from the Red-browed Amazon parrot (Amazona rhodocorytha) kept at a conservation institute in Brazil. High genetic variability in virulence was observed, since 12 virulence profiles were found among 14 strains. The number of virulence-associated genes of single strains ranged from 5 to 22 out of 33 genes tested, and only one strain did not present any virulence genes. Regarding adhesion genes, most strains presented from two to five genes, and crlA (85.7%) and fimC (85.7%) were the most frequent. Frequencies were similar for invasion and iron acquisition genes. Variations among genes were observed for serum resistance and toxin-related genes. Some of the E. coli strains isolated from parrots presented virulence genes that are commonly associated with pathotypes of human origin, including newborn meningitis E. coli, uropathogenic E. coli, and sepsis-associated E. coli. It is noteworthy that some of these genes were present in the majority of the analyzed strains. Our results indicate that these strains detected in clinically healthy parrots can be potential reservoirs of several virulence-associated genes. These genes can be transmitted to other E. coli strains, including those that affect humans. These E. coli strains present a high pathogenic potential of virulence-associated genes in extraintestinal pathogenic E. coli strains.(AU)

Development of copper hatch baskets and evaluation of the productive indexes of a commercial hatchery

Background: The dissemination of pathogenic microorganisms in hatcheries leads to a higher number of contaminated eggs, causing reduction in hatchability and increase of discarded chicks. Sanitation programs are crucial for maximum hatchability and chick quality. Efforts have been made to find alternative approaches to the conventional disinfectants, and surfaces with copper, which have antimicrobial properties, could assist in this process. However, the possible adverse effects of copper surfaces on chicks in hatcheries have not yet been evaluated. The present study aimed at developing hatch baskets composed of copper and evaluating the effect of these baskets on the productive indexes of a hatchery. Materials, Methods & Results: For this experiment, 3.15 kg hatch tray prototypes with 99.9% Cu (Cu11000) were developed to fit inside conventional polypropylene hatch baskets (580 × 755 × 83 mm). Six polypropylene hatch baskets (control group) and six polypropylene hatch baskets covered by 99.9% copper (Cu11000) hatch trays (test group) were evaluated during 5 hatchings. Hatched eggs and chicks remained in contact with the hatch basket surfaces for at least 72 h, corresponding to the entire period in which they were located in the hatcher. Cleaning and disinfection programs of the hatchery were not modified. The level of microbial contamination on the hatch baskets was evaluated at 6 different periods: 0 h (initial contamination after disinfection and egg transfer to the trays); 24 h, 30 h, 45 h and 60 h after the first sampling; and at the moment when chicks were removed from the hatching cabinet and transferred to the chick-holding room (> 60 h). Counting of total moulds and yeasts, mesophilic microorganisms, Enterobacteria and Escherichia coli colonies was performed. The number of hatched chicks, non-hatched eggs, and chicks discarded were registered for each hatching. Microbiologic analyses showed no growth on hatch baskets neither of the..

Detection of virulence genes in Salmonella Heidelberg isolated from chicken carcasses.

During the last years, Brazilian government control programs have detected an increase of Salmonella Heidelberg in poultry slaughterhouses a condition that poses a threat to human health However, the reasons remain unclear. Differences in genetic virulence profiles may be a possible justification. In addition, effective control of Salmonella is related to an efficient epidemiological surveillance system through genotyping techniques. In this context, the aim of this study was the detection of 24 virulence-associated genes in 126 S. Heidelberg isolates. We classified the isolates into 56 different genetic profiles. None of the isolates presented all the virulence genes. The prevalence of these genes was high in all tested samples as the lowest number of genes detected in one isolate was 10/24. The lpfA and csgA (fimbriae), invA and sivH (TTSS), and msgA and tolC (intracellular survival) genes were present in 100% of the isolates analyzed. Genes encoding effector proteins were detected in the majority of SH isolates. No single isolate had the sefA gene. The pefA gene was found in only four isolates. We have also performed a screening of genes associated with iron metabolism: 88.9% of isolates had the iroN geneand 79.4% the sitC gene . Although all the isolates belong to the same serotype, several genotypic profiles were observed. These findings suggest that there is a diversity of S. Heidelberg isolates in poultry products. The fact that a single predominant profile was not found in this study indicates the presence of variable sources of contamination caused by SH. The detection of genetic profiles of Salmonella strains can be used to determine the virulence patterns of SH isolates.

Biofilm formation by Salmonella Enteritidis at different incubation temperatures

Background: The genus Salmonella, associated with poultry products, is considered the leading cause of foodborne outbreaks in humans in many countries. In Brazil, Salmonella Enteritidis (SE) is the serovar remains as one most frequentlyisolated from humans, and it is also a major serovar found in animals, food, animal feed, and environmental samples, despiteall the efforts to control this pathogen. Also, the bacterium is able to form biofilms on different surfaces, protecting cellsfrom both cleaning and sanitizing procedures in the food industries. This study aimed to verify the ability of SalmonellaEnteritidis isolates to form biofilm on polystyrene at different incubation temperatures.Materials, Methods & Results: A total of 171 SE samples were isolated from foodborne outbreaks (foods and stool cultures) and poultry products between 2003 and 2010. The biofilm-forming ability of samples was measured at four differenttemperatures (3°C, 9ºC, 25ºC, and 36ºC), for 24 h, simulating temperatures usually found in poultry slaughterhouses. Later,200 μL of each bacterial suspension was inoculated, in triplicate, onto 96-well, flat-bottomed sterile polystyrene microtiterplates, washed, after that, the biofilm was fixed with methanol. The plates were dried at ambient temperature, stained with2% Huckers crystal violet. Afterwards, absorbance was read using an ELISA plate reader and the optical density (OD)of each isolate was obtained by the arithmetic mean of the absorbance of three wells and this value was compared withthe mean absorbance of negative controls (ODnc). The following classification was used for the determination of biofilmformation: no biofilm production, weak biofilm production, moderate biofilm production and strong biofilm production.Results demonstrated all isolates from stool cultures and foods involved in foodborne outbreaks, at least one of the fourtemperatures tested, were able to form biofilm...(AU)

Salmonella Enteritidis forms biofilm under low temperatures on different food industry surfaces / Salmonella Enteritidis forma biofilme sob baixas temperaturas em diferentes superfícies da indústria de alimentos

We evaluated the influence of temperature on the ability of Salmonella Enteritidis (SE) to form biofilms on stainless steel, polyethylene, and polyurethane surfaces under different hygiene procedures. These materials were placed on SE culture and incubated at 42±1 ºC, 36±1 ºC, 25±1 ºC, 9±1 ºC, and 3±1 ºC for 4, 8, 12, and 24 h. Hot water at 45 ºC and 85 ºC, 0.5% peracetic acid solution, and 1% quaternary ammonia were used for hygienization. Biofilm formation occurred at all temperatures evaluated, highlighting at 3 ºC which has not been reported as an ideal temperature for the adhesion of SE to these materials. The SE adhered more often to polyethylene surfaces than to polyurethane and stainless steel surfaces (P<0.05). Peracetic acid and water at 85 ºC had similar hygienization efficiency (P<0.05) followed by quaternary ammonia whereas water at 45 ºC was not effective. SE adhered to these materials under low temperatures which to date have been deemed safe for food preservation.(AU)

Avaliou-se o efeito da temperatura na capacidade de Salmonella Enteritidis (SE) formar biofilme em superfícies de aço inoxidável, polietileno e poliuretano e diferentes processos de higienização. Corpos de prova destes materiais foram postos frente a culturas de SE e incubados a 42±1 ºC, 36±1 ºC, 25±1 ºC, 9±1 ºC e 3±1 ºC por 4, 8, 12 e 24 horas. Para a higienização foram testados água aquecida a 45ºC e 85 ºC e soluções de ácido peracético 0,5% e amônia quaternária 1%. Verificou-se a formação de biofilmes em todas as temperaturas avaliadas, ressaltando-se a 3 ºC, ainda não citada como propícia para adesão de SE. Houve maior adesão ao polietileno do que ao poliuretano e ao aço inoxidável (P<0.05). Para higienização, o ácido peracético e a água a 85 ºC tiveram ação semelhante (P<0.05), seguidos por amônia quaternária, enquanto que a água a 45 ºC não foi eficaz. Todos os materiais avaliados propiciaram a aderência de SE, mesmo sob temperaturas baixas, consideradas até então seguras para a conservação dos alimentos.(AU)

Biofilm formation by Salmonella Enteritidis at different incubation temperatures

Background: The genus Salmonella, associated with poultry products, is considered the leading cause of foodborne outbreaks in humans in many countries. In Brazil, Salmonella Enteritidis (SE) is the serovar remains as one most frequentlyisolated from humans, and it is also a major serovar found in animals, food, animal feed, and environmental samples, despiteall the efforts to control this pathogen. Also, the bacterium is able to form biofilms on different surfaces, protecting cellsfrom both cleaning and sanitizing procedures in the food industries. This study aimed to verify the ability of SalmonellaEnteritidis isolates to form biofilm on polystyrene at different incubation temperatures.Materials, Methods & Results: A total of 171 SE samples were isolated from foodborne outbreaks (foods and stool cultures) and poultry products between 2003 and 2010. The biofilm-forming ability of samples was measured at four differenttemperatures (3°C, 9ºC, 25ºC, and 36ºC), for 24 h, simulating temperatures usually found in poultry slaughterhouses. Later,200 μL of each bacterial suspension was inoculated, in triplicate, onto 96-well, flat-bottomed sterile polystyrene microtiterplates, washed, after that, the biofilm was fixed with methanol. The plates were dried at ambient temperature, stained with2% Hucker’s crystal violet. Afterwards, absorbance was read using an ELISA plate reader and the optical density (OD)of each isolate was obtained by the arithmetic mean of the absorbance of three wells and this value was compared withthe mean absorbance of negative controls (ODnc). The following classification was used for the determination of biofilmformation: no biofilm production, weak biofilm production, moderate biofilm production and strong biofilm production.Results demonstrated all isolates from stool cultures and foods involved in foodborne outbreaks, at least one of the fourtemperatures tested, were able to form biofilm...

Salmonella Enteritidis forms biofilm under low temperatures on different food industry surfaces / Salmonella Enteritidis forma biofilme sob baixas temperaturas em diferentes superfícies da indústria de alimentos

ABSTRACT: We evaluated the influence of temperature on the ability of Salmonella Enteritidis (SE) to form biofilms on stainless steel, polyethylene, and polyurethane surfaces under different hygiene procedures. These materials were placed on SE culture and incubated at 42±1 ºC, 36±1 ºC, 25±1 ºC, 9±1 ºC, and 3±1 ºC for 4, 8, 12, and 24 h. Hot water at 45 ºC and 85 ºC, 0.5% peracetic acid solution, and 1% quaternary ammonia were used for hygienization. Biofilm formation occurred at all temperatures evaluated, highlighting at 3 ºC which has not been reported as an ideal temperature for the adhesion of SE to these materials. The SE adhered more often to polyethylene surfaces than to polyurethane and stainless steel surfaces (P<0.05). Peracetic acid and water at 85 ºC had similar hygienization efficiency (P<0.05) followed by quaternary ammonia whereas water at 45 ºC was not effective. SE adhered to these materials under low temperatures which to date have been deemed safe for food preservation.

RESUMO: Avaliou-se o efeito da temperatura na capacidade de Salmonella Enteritidis (SE) formar biofilme em superfícies de aço inoxidável, polietileno e poliuretano e diferentes processos de higienização. Corpos de prova destes materiais foram postos frente a culturas de SE e incubados a 42±1 ºC, 36±1 ºC, 25±1 ºC, 9±1 ºC e 3±1 ºC por 4, 8, 12 e 24 horas. Para a higienização foram testados água aquecida a 45ºC e 85 ºC e soluções de ácido peracético 0,5% e amônia quaternária 1%. Verificou-se a formação de biofilmes em todas as temperaturas avaliadas, ressaltando-se a 3 ºC, ainda não citada como propícia para adesão de SE. Houve maior adesão ao polietileno do que ao poliuretano e ao aço inoxidável (P<0.05). Para higienização, o ácido peracético e a água a 85 ºC tiveram ação semelhante (P<0.05), seguidos por amônia quaternária, enquanto que a água a 45 ºC não foi eficaz. Todos os materiais avaliados propiciaram a aderência de SE, mesmo sob temperaturas baixas, consideradas até então seguras para a conservação dos alimentos.

ATP-Bioluminescence and conventional microbiology for hygiene evaluation of cutting room surfaces in poultry slaughterhouse

Background: The hygiene procedures in poultry slaughterhouses consist in the use of hot water, detergent and sanitizing, configuring Sanitation Standard Operating Procedure (SSOP). These actions control contamination in food processing environments, especially by pathogenic microorganisms, which cause diseases with impact on public health and economic losses. The microbiological control of aerobic mesophiles, Staphylococcus aureus and Escherichia coli, are used as indicators of contamination. The hygienic-sanitary conditions on the surfaces of the poultry slaughterhouse cuttting room were evaluated, before and after cleaning and sanitizing procedures.Materials, Methods & Results: Conventional microbiology (Rodac plates and sponge for quantification of aerobic mesophiles, Staphylococcus aureus and Escherichia coli) and ATP-Bioluminescence were used to analyze the action of hot water and the active principles peracetic acid, quaternary ammonia and biguanide in the standard pre-operational hygiene procedure in the cutting room of the poultry slaughterhouse under Federal Inspection with slaughter capacity of more than 20.000 birds/h. The evaluations were performed on three lines of chicken thigh cuts at the same time and in a completely randomized manner on stainless steel surfaces, polyurethane belts and polyethylene boards. Samples were made in four replicates at the three surface totaling 108 assay for each microorganism. The samples were collected at the end of the cutting process, before and after washing the surfaces with hot water (between 45 and 50ºC) and after sanitization with 0.5% peracetic acid, 2% quaternary ammonia and 1% biguanide. The ATP-Bioluminescence method detected organic matter at all collected points and Rodac plates allowed a better recovery of microorganisms than sponges for quantification of aerobic mesophiles, E. coli and S. aureus.[...](AU)

ATP-Bioluminescence and conventional microbiology for hygiene evaluation of cutting room surfaces in poultry slaughterhouse

Background: The hygiene procedures in poultry slaughterhouses consist in the use of hot water, detergent and sanitizing, configuring Sanitation Standard Operating Procedure (SSOP). These actions control contamination in food processing environments, especially by pathogenic microorganisms, which cause diseases with impact on public health and economic losses. The microbiological control of aerobic mesophiles, Staphylococcus aureus and Escherichia coli, are used as indicators of contamination. The hygienic-sanitary conditions on the surfaces of the poultry slaughterhouse cuttting room were evaluated, before and after cleaning and sanitizing procedures.Materials, Methods & Results: Conventional microbiology (Rodac plates and sponge for quantification of aerobic mesophiles, Staphylococcus aureus and Escherichia coli) and ATP-Bioluminescence were used to analyze the action of hot water and the active principles peracetic acid, quaternary ammonia and biguanide in the standard pre-operational hygiene procedure in the cutting room of the poultry slaughterhouse under Federal Inspection with slaughter capacity of more than 20.000 birds/h. The evaluations were performed on three lines of chicken thigh cuts at the same time and in a completely randomized manner on stainless steel surfaces, polyurethane belts and polyethylene boards. Samples were made in four replicates at the three surface totaling 108 assay for each microorganism. The samples were collected at the end of the cutting process, before and after washing the surfaces with hot water (between 45 and 50ºC) and after sanitization with 0.5% peracetic acid, 2% quaternary ammonia and 1% biguanide. The ATP-Bioluminescence method detected organic matter at all collected points and Rodac plates allowed a better recovery of microorganisms than sponges for quantification of aerobic mesophiles, E. coli and S. aureus.[...]

Evaluation of the effectiveness and safety of a genetically modified live vaccine in broilers challenged with Salmonella Heidelberg.

Salmonellosis ranks among the major diseases of commercial poultry, and its presence in poultry flocks is responsible for economic losses and risks related to public health. Vaccines are an important tool within integrated programmes to control salmonellosis. The purpose of this study was to assess cross-protection provided by the Poulvac® ST vaccine in the control of Salmonella Heidelberg in experimentally challenged 3- and 21-day-old birds. Eighty birds were identified and separated into four treatments (T1: vaccinated and challenged at 3 days of age, T2: unvaccinated and challenged at 3 days of age, T3: vaccinated and challenged at 21 days of age, and T4: unvaccinated and challenged at 21 days of age). The inoculum was produced from a Brazilian field strain of SH. At the end of the experiment, caecum and liver/spleen samples were collected for quantitative and qualitative analysis of SH, respectively. Analysis of the liver/spleen showed that Poulvac® ST significantly (P ≤ 0.05) reduced the percentage of SH positivity in the group challenged at 3 days of age, while in the group challenged at 21 days this difference was almost considered significant (P = 0.1818). On the other hand, there was no statistically significant difference in SH count in the caecum (CFU/g) in the group challenged at 3 days, but for the group challenged at 21 days the SH counts were significantly (P ≤ 0.05) lower in the vaccinated group when compared to the positive control.

Classificação de Escherichia coli patogênica aviária (APEC)e de Escherichia coli uropatogênica (UPEC) em grupos filogenéticose associação com a patogenicidade in vivo / Classification of Avian Pathogenic Escherichia coli (APEC) and Human Uropathogenic Escherichia coli (UPEC) in phylogenetic groups and association with pathogenicity in vivo

Background: Avian pathogenic E. coli (APEC) and uropathogenic E. coli (UPEC) are responsible, respectively, for avian colibacillosis and for 80% of urinary tract infections in humans. E. coli control is difficult due to the absence of a reliable method to differentiate pathogenic and commensal strains. Genetic similarity between APEC and UPEC suggests a common ancestral origin and the capability of potentially pathogenic strains to affect human health. The classification in phylogenetic groups facilitates the identification of pathogenic strains. The objective of this work was to classify APEC and UPEC E. coli strains into phylogenetic groups and to associate it with in vivo pathogenicity. Materials, Methods & Results: 460 APEC and 450 UPEC strains, stored in BHI with glycerol at -80°C, were selected. APEC strains were isolated from cellulitis, respiratory tract and poultry litter of broiler flocks from Southern Brazil. The UPEC strains from urinary tract infection were provided by a hospital in Porto Alegre. After DNA extraction, APEC and UPEC strains were classified into four phylogenetic groups (A, B1, B2 and D) by a multiplex-PCR protocol for the detection of the chuA and yjaA genes and the TspE4.C2 DNA fragment. Phylogenetic groups were associated with pathogenicity indexes (PI), presented on a scale of 0 to 10, which were previously obtained through the inoculation of APEC strains in one-day-old chicks. Phylogenetic groups were also associated with the presence of 38 virulence-associated genes. The multiplex-PCR protocol was able to differentiate 100% of the APEC and UPEC strains in the four phylogenetic groups. The majority of APEC strains were classified into phylogenetic groups D (31.1%) and B2 (24.1%). On the other hand, the majority of UPEC strains were classified into B2 (53.6%). Among APEC strains, five genes (crl, mat, ompA, fimC and fimH) […](AU)

Classificação de Escherichia coli patogênica aviária (APEC)e de Escherichia coli uropatogênica (UPEC) em grupos filogenéticose associação com a patogenicidade in vivo / Classification of Avian Pathogenic Escherichia coli (APEC) and Human Uropathogenic Escherichia coli (UPEC) in phylogenetic groups and association with pathogenicity in vivo

Background: Avian pathogenic E. coli (APEC) and uropathogenic E. coli (UPEC) are responsible, respectively, for avian colibacillosis and for 80% of urinary tract infections in humans. E. coli control is difficult due to the absence of a reliable method to differentiate pathogenic and commensal strains. Genetic similarity between APEC and UPEC suggests a common ancestral origin and the capability of potentially pathogenic strains to affect human health. The classification in phylogenetic groups facilitates the identification of pathogenic strains. The objective of this work was to classify APEC and UPEC E. coli strains into phylogenetic groups and to associate it with in vivo pathogenicity. Materials, Methods & Results: 460 APEC and 450 UPEC strains, stored in BHI with glycerol at -80°C, were selected. APEC strains were isolated from cellulitis, respiratory tract and poultry litter of broiler flocks from Southern Brazil. The UPEC strains from urinary tract infection were provided by a hospital in Porto Alegre. After DNA extraction, APEC and UPEC strains were classified into four phylogenetic groups (A, B1, B2 and D) by a multiplex-PCR protocol for the detection of the chuA and yjaA genes and the TspE4.C2 DNA fragment. Phylogenetic groups were associated with pathogenicity indexes (PI), presented on a scale of 0 to 10, which were previously obtained through the inoculation of APEC strains in one-day-old chicks. Phylogenetic groups were also associated with the presence of 38 virulence-associated genes. The multiplex-PCR protocol was able to differentiate 100% of the APEC and UPEC strains in the four phylogenetic groups. The majority of APEC strains were classified into phylogenetic groups D (31.1%) and B2 (24.1%). On the other hand, the majority of UPEC strains were classified into B2 (53.6%). Among APEC strains, five genes (crl, mat, ompA, fimC and fimH) […]

Use of Molecular Pathogenicity Indices to Identify Pathogenic Strains of Pasteurella multocida.

In addition to being the causative agent of fowl cholera (FC), Pasteurella multocida is also one of the most prevalent opportunistic pathogens associated with respiratory diseases in various hosts. However, understanding of the traits that distinguish the virulent isolates that cause FC is still limited. The objective of this study was to characterize P. multocida isolates of Brazil by PCR-restriction fragment length polymorphism (PCR-RFLP) analysis in order to determine if strain-type correlates with virulence or with 22 previously studied virulence genes. The PCR-RFLP was used to classify the isolates into seven strain types, and the isolates in Profile II had a higher pathogenicity index (P < 0.05) than did those in Profiles I, V, and VI. The overall identity among the nucleotide sequences of the ompH was 89.8%. Furthermore, strains available in GenBank showed a high level of homology of the different bacterial serotypes with the groupings resulting from the PCR-RFLP. Strain Types I and II showed the highest identity with Serotypes 3 (100%) and 3-4 (99.1%), respectively. Detection of the pfhA gene indicated the presence of strains that are highly pathogenic. The screening detection of 22 virulence genes and inference through the decision tree models comparing the results of pathogenicity indices permitted the identification of the most highly pathogenic strains of P. multocida .

Campylobacter in broiler slaughter samples assessed by direct count on mCCDA and Campy-Cefex agar

ABSTRACT Campylobacter spp. cause foodborne illnesses in humans primarily through the consumption of contaminated chicken. The aim of this study was to evaluate the United States Department of Agriculture's (USDA) recommended methodology, protocol MLG 41.02, for the isolation, identification and direct plate counting of Campylobacter jejuni and C. coli samples from the broiler slaughtering process. A plating method using both mCCDA and Campy-Cefex agars is recommended to recover Campylobacter cells. It is also possible to use this method in different matrices (cloacal swabs and water samples). Cloacal swabs, samples from pre-chiller and post-chiller carcasses and samples of pre-chiller, chiller and direct supply water were collected each week for four weeks from the same flock at a slaughterhouse located in an abattoir in southern Brazil. Samples were analyzed to directly count Campylobacter spp., and the results showed a high frequency of Campylobacter spp. on Campy-Cefex agar. For the isolated species, 72% were identified as Campylobacter jejuni and 38% as Campylobacter coli. It was possible to count Campylobacter jejuni and Campylobacter coli from different samples, including the water supply samples, using the two-agar method. These results suggest that slaughterhouses can use direct counting methods with both agars and different matrices as a monitoring tool to assess the presence of Campylobacter bacteria in their products.

Campylobacter in broiler slaughter samples assessed by direct count on mCCDA and Campy-Cefex agar

Campylobacter spp. cause foodborne illnesses in humans primarily through the consumption of contaminated chicken. The aim of this study was to evaluate the United States Department of Agriculture's (USDA) recommended methodology, protocol MLG 41.02, for the isolation, identification and direct plate counting of Campylobacter jejuni and C. coli samples from the broiler slaughtering process. A plating method using both mCCDA and Campy-Cefex agars is recommended to recover Campylobacter cells. It is also possible to use this method in different matrices (cloacal swabs and water samples). Cloacal swabs, samples from pre-chiller and post-chiller carcasses and samples of pre-chiller, chiller and direct supply water were collected each week for four weeks from the same flock at a slaughterhouse located in an abattoir in southern Brazil. Samples were analyzed to directly count Campylobacter spp., and the results showed a high frequency of Campylobacter spp. on Campy-Cefex agar. For the isolated species, 72% were identified as Campylobacter jejuni and 38% as Campylobacter coli. It was possible to count Campylobacter jejuni and Campylobacter coli from different samples, including the water supply samples, using the two-agar method. These results suggest that slaughterhouses can use direct counting methods with both agars and different matrices as a monitoring tool to assess the presence of Campylobacter bacteria in their products.(AU)

SEROVARS AND ANTIMICROBIAL RESISTANCE OF Salmonella spp. ISOLATED FROM TURKEY AND BROILER CARCASSES IN SOUTHERN BRAZIL BETWEEN 2004 AND 2006.

Salmonella spp. causes diseases in fowls, when species-specific serovars (Salmonella Pullorum and S.Gallinarum) are present in flocks, and public health problems, when non-typhoid serovars are isolated, as well as possible bacterial resistance induced by the preventive and therapeutic use of antimicrobials in animal production. This study describes the serovars and bacterial resistance of 280 Salmonella spp. strains isolated from turkey and broiler carcasses in Southern Brazil between 2004 and 2006. Salmonella Enteritidis was the most prevalent serovar (55.7%), followed by Heidelberg (5.0%), Agona (4.3%), Bredeney (3.9%), Hadar (3.2%), and Typhimurium (2.9%). Tennessee and S. Enterica subspecies enterica(O: 4.5) were isolated only in turkeys, and Hadar (18.6%) was the most prevalent serovar in this species. Antimicrobial susceptibility tests were performed in 178 isolates (43 from turkeys and 135 from broilers). All isolates were sensitive to amoxicillin + clavulanic acid, polymyxin B, ciprofloxacin, and norfloxacin, and were resistant to bacitracin and penicillin. Broiler carcass isolates showed resistance to nalidixic acid (48.9%), nitrofurantoin (34.3%), neomycin (9.6%), tetracycline (5.2%), and kanamycin (8.9%); and turkey carcass isolates were resistant to nalidixic acid (62.8%), tetracycline (34.9%), and neomycin (30.2%), with a significant difference in turkeys when compared to broiler carcass isolates. These results indicate the need for judicious use of antimicrobials in livestock production, given that the serovars identified are potential causes of food poisoning.