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This study aimed to analyze 19 isolates of Salmonella spp., from broiler litter swabs in the State of São Paulo, by typing and analyzing the detection of resistance genes associated with ESBL's (Extended Spectrum Beta Lactamase), AMPC (C-type cephalosporinases) and carbapenemases by molecular and phenotypic techniques. A PCR microarray platform (Check and Trace by Check-Points) was used to identify the isolated serotype. The isolates were also evaluated for identification of carbapenemase genes, MCR 1-2 (colistin resistance), AmpC (C-type cephalosporins), and ESBLs (ß-lactamases resistance). To identify phenotypic antibiotic resistance, the minimal inhibitory concentration (MIC) was evaluated with the antibiotics meropenem, amoxicillin, and ceftriaxone. The most prevalent serotypes identified were S. Infantis and S. Saintpaul, with a prevalence of 15.07% (3/19). Other strains identified were S. Cerro, S. Sandiego, S. Kentucky, S. Alachua, S. Javiana, S. Livingstone, S. Typhimurium, S. Heidelberg, non-enteric Salmonella, and a Salmonella not typifiable by the typing kit. All samples were negative for identifying carba resistance genes, MCR, ESBL, and AmpC. In the phenotypic profile, meropenem was the least resistant, while amoxicillin and ceftriaxone showed a high resistance pattern. The results show that phenotypic resistance is not associated with the presence of resistance genes studied here. In addition, the resistant bacteria found in MIC have resistance mechanisms not associated with the genes studied here. Additional measures must be implemented to prevent the indiscriminate use of antimicrobial agents therapeutically or as growth promoters.(AU)

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The use of chicken embryos (CEs) as an in vivo experimental model for different pharmaceutical purposes is not a novelty. However, in recent years, the number of reports employing CE to evaluate several parameters, such as the toxicity and efficacy of drugs and/or nanosystems, has increased. Therefore, this review discusses the relevance of CE for drug testing, emphasizing the inoculation routes and the embryonic stages. The challenges to be overcome, as well as some practical recommendations to allow CE to be more explored as a promising in vivo model in drug analyses, are also highlighted.

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Raising backyard birds is a common practice in Brazil, mainly in the countryside or suburban areas. However, the level of respiratory pathogens in these animals is unknown. We sampled two hundred chickens from 19 backyard flocks near commercial poultry farms and performed ELISA to Infectious Bronchitis Virus, avian Metapneumovirus, Mycoplasma synoviae and Mycoplasma gallisepticum. We evaluated the association between the predictive ability of ELISA and Hemagglutination-inhibition (HI)by comparing results from eight flocks positive to Mycoplasma gallisepticum on ELISA. Besides, we assessed essential biosecurity measures in the properties (multiple species birds, rodent control, hygienic conditions, and water quality for the bird`s consumption). We could access the vaccination program only on four properties; in three of them, the birds were supposedly vaccinated for IBV. Overall the properties had a poor score for the biosecurity measures, and the seroprevalence in backyard poultry flocks for IBV, a MPV, MS, and MG were respectively 87.5% (14/16), 89.5% (17/19), 100 (19/19) and MG 84.21% (16/19). We found low specificity and predictive value between ELISA and HI in MG analysis and a positive correlation between the presence of clinical symptoms and mean MG titers. Backyard chicken are pathogens reservoirs and pose a risk for the commercial poultry farms in the region, and further efforts of the governmental entities and private sector of poultry production should consider these information to avoid future economic losses.(AU)

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Raising backyard birds is a common practice in Brazil, mainly in the countryside or suburban areas. However, the level of respiratory pathogens in these animals is unknown. We sampled two hundred chickens from 19 backyard flocks near commercial poultry farms and performed ELISA to Infectious Bronchitis Virus, avian Metapneumovirus, Mycoplasma synoviae and Mycoplasma gallisepticum. We evaluated the association between the predictive ability of ELISA and Hemagglutination-inhibition (HI)by comparing results from eight flocks positive to Mycoplasma gallisepticum on ELISA. Besides, we assessed essential biosecurity measures in the properties (multiple species birds, rodent control, hygienic conditions, and water quality for the bird`s consumption). We could access the vaccination program only on four properties; in three of them, the birds were supposedly vaccinated for IBV. Overall the properties had a poor score for the biosecurity measures, and the seroprevalence in backyard poultry flocks for IBV, a MPV, MS, and MG were respectively 87.5% (14/16), 89.5% (17/19), 100 (19/19) and MG 84.21% (16/19). We found low specificity and predictive value between ELISA and HI in MG analysis and a positive correlation between the presence of clinical symptoms and mean MG titers. Backyard chicken are pathogens reservoirs and pose a risk for the commercial poultry farms in the region, and further efforts of the governmental entities and private sector of poultry production should consider these information to avoid future economic losses.

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1. The aim of this study was to analyse the association between Escherichia coli isolates recovered from turkeys and the expression of beta-lactamase genes, such as extended spectrum beta-lactamase (ESBL) and ampicillin class C (AmpC). The phenotype of the resistance profile was examined using the association between amoxicillin and ceftiofur resistance. 2. Results showed that 84% from the turkey isolates harboured 4 or 5 genes associated with the CoIV plasmid. In an antibiogram test, 82% of the isolates were multidrug-resistant, the highest levels of resistance being against erythromycin (99%) and amoxicillin (76.1%). ESBL-positive groups were 31% positive for the ctx-m-2 gene, 6.8% were positive for ctx-m-8 and 70% harboured the tem wild gene. 3. All positive isolates from the AmpC beta-lactamase-positive group harboured the cmy-2 gene. The presence of the cmy-2 gene was associated with both the CTX-group genes and resistance to ceftiofur. 4. There was a high prevalence of avian pathogenic E. coli in suspected cases of colibacillosis in turkeys and a high antimicrobial resistance index. The results highlighted the risk of ceftiofur resistance and the presence of both ESBL and AmpC beta-lactamase E. coli in the turkey production chain.

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1. The aim of the present study was to determine if the 9R-strain of the Salmonella Gallinarum live vaccine was responsible for having fowl typhoid outbreaks in chicken flocks from both chicken and turkey breeders as well as to verify the antimicrobial resistance of the isolates from the outbreaks. 2. The triplex polymerase chain reaction, standard antimicrobial test, beta-lactamase genes identification and Ion Torrent PMG whole-genome sequence were used in the field isolates and in the vaccine strain of S. Gallinarum. 3. The 60 tested isolates were not from vaccine origin and manifested high resistance to drugs from macrolide and quinolone groups. Whole-genome sequencing (WGS) and single nucleotide polymorphism analysis on selected isolates for core genes from Salmonella enterica confirmed the wild origin of these isolates and showed two possible sources of S. Gallinarum in the studied outbreaks. 4. S. Gallinarum isolated from fowl typhoid outbreaks in the studied period were not caused by the use of the SG9R live vaccine. The source of strains sequenced was diverse.

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Biofilms are frequently related to invasive fungal infections and are reported to be more resistant to antifungal drugs than planktonic cells. The structural complexity of the biofilm as well as the presence of a polymeric extracellular matrix (ECM) is thought to be associated with this resistant behavior. Scanning electron microscopy (SEM) after room temperature glutaraldehyde-based fixation, have been used to study fungal biofilm structure and drug susceptibility but they usually fail to preserve the ECM and, therefore, are not an optimised methodology to understand the complexity of the fungal biofilm. Thus, in this work, we propose a comparative analysis of room-temperature and cryofixation/freeze substitution of Candida albicans biofilms for SEM observation. Our experiments showed that room-temperature fixative protocols using glutaraldehyde and osmium tetroxide prior to alcohol dehydration led to a complete extraction of the polymeric ECM of biofilms. ECM from fixative and alcohol solutions were recovered after all processing steps and these structures were characterised by biochemistry assays, transmission electron microscopy and mass spectrometry. Cryofixation techniques followed by freeze-substitution lead to a great preservation of both ECM structure and C. albicans biofilm cells, allowing the visualisation of a more reliable biofilm structure. These findings reinforce that cryofixation should be the indicated method for SEM sample preparation to study fungal biofilms as it allows the visualisation of the EMC and the exploration of the biofilm structure to its fullest, as its structural/functional role in interaction with host cells, other pathogens and for drug resistance assays.

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1. The aim was to determine the importance of a contaminated diet as a possible cause of Campylobacter jejuni infection in broilers. 2. This study evaluated the viability of C. jejuni in both starter and finisher diets and the interference from other mesophilic bacteria in this viability. 3. Starter and finisher samples of broiler diet were deliberately contaminated with 3 or 5 log CFU·g-1 of C. jejuni (NCTC 11351) and then maintained at two different storage temperatures (25°C or 37°C) for 3 or 5 d. 4. C. jejuni survived during this period and, when inoculated at 103 CFU·g-1, multiplied with greater proliferation at a storage temperature of 37°C. There was no relationship between the amount of mesophilic bacteria and C. jejuni viability. 5. This study highlights the importance of the diet in the epidemiology of C. jejuni in broilers.

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Campylobacteriosis is a worldwide foodborne zoonosis disease caused by Campylobacter jejuni. This microorganism is considered a commensal bacterium in chicken hosts. C. jejuni produces epithelial cell modifications and induces a cytokine gene transcription innate immunity repertoire. In the present study, we describe the invasiveness, morphological cellular modifications, and transcript level expressions of innate immune cytokines from C. jejuni-inoculated chicken ileum explants. C. jejuni was internalized by epithelial ileum cells at 15 minutes postinoculation (p.i.) and was detected intracellularly for 4hs (p.i.). Inoculated explants displayed significant increases in cell height. C. jejuni induced a significant elevation of Transforming Growth Factor Beta 3 (TGF-b3) and Interleukin-1b (IL-1b) transcripts. In conclusion, C. jejuni is internalized in explanted epithelial ileum cells, produces morphological cell modifications, and induces gene transcription of both anti-inflammatory and pro-inflammatory cytokines.

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Campylobacteriosis is a worldwide foodborne zoonosis disease caused by Campylobacter jejuni. This microorganism is considered a commensal bacterium in chicken hosts. C. jejuni produces epithelial cell modifications and induces a cytokine gene transcription innate immunity repertoire. In the present study, we describe the invasiveness, morphological cellular modifications, and transcript level expressions of innate immune cytokines from C. jejuni-inoculated chicken ileum explants. C. jejuni was internalized by epithelial ileum cells at 15 minutes postinoculation (p.i.) and was detected intracellularly for 4hs (p.i.). Inoculated explants displayed significant increases in cell height. C. jejuni induced a significant elevation of Transforming Growth Factor Beta 3 (TGF-b3) and Interleukin-1b (IL-1b) transcripts. In conclusion, C. jejuni is internalized in explanted epithelial ileum cells, produces morphological cell modifications, and induces gene transcription of both anti-inflammatory and pro-inflammatory cytokines.(AU)

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The serum biochemical profiles, thyroid hormones, body weights and the production and quality of eggs subsequent to moulting, were compared in laying hens subjected to conventional forced moulting or forced moulting with a diet high in zinc. A total of 200 Dekalb White laying hens in their second production cycle were studied. Blood sampling was conducted in a factorial experimental design (2 × 3) with two methods of moulting (fasting or zinc) and three sampling periods (pre-moult, moult and subsequent peak). Total egg protein content, including globulins, was greater with the zinc diet, whereas egg weight and albumen percentage were greater after fasting. The zinc method resulted in an increased shell thickness and calcium percentage but lower percentage of phosphorus. During the moulting period, the hens in the zinc group had heavier mean body weights. It was concluded that moulting with a high-zinc diet could replace fasting, without negative effects on body weight, biochemical variables or subsequent egg quality and production. The zinc method was also better for the birds' welfare.

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A diversificação da produção industrial de alimentos de origem suína e o intercâmbio comercial de animais e seus derivados destinados ao consumo humano podem ser importantes disseminadores de sorovares de Salmonella spp. na cadeia alimentar. Objetivou-se avaliar em 86 cepas de Salmonella spp., isoladas em granja de terminação e no abate de suínos, a ocorrência de três genes de virulência (invA, agfA e lpfA), bem como a similaridade genética entre elas. A ocorrência do gene invA foi verificada em 100% das amostras. O gene lpfA foi detectado em 80,23% (69/86) das cepas, não foi detectado em S. Panama e estava presente em todas as cepas de S. Infantis. O gene agfA foi detectado em 63,95% (55/86) das amostras. S. Agona apresentou positividade para todos os genes de virulência estudados. A análise de homologia entre as cepas agrupou os diferentes sorovares em clusters. A similaridade foi independente do local de isolamento, o que demonstra a presença de clones ao longo da cadeia de produção e a existência de multiplicidade de fontes para a infecção dos animais, como a ração, e a contaminação cruzada das carcaças. A pesquisa de genes de virulência e a avaliação da proximidade gênica permitem a caracterização e um maior entendimento sobre cepas de Salmonella circulantes na cadeia produtiva de suínos e, assim, podem subsidiar medidas de controle durante o processo produtivo com o objetivo de garantir a saúde do consumidor...

The diversification of industrial food production of swine origin and trade of animals and their derivatives for human consumption may be important disseminators of serovars of Salmonella spp. in the food chain. This study aimed to evaluate 86 strains of Salmonella spp. isolated form in the finishing and slaughter of pigs, the occurrence of three virulence genes (invA, agfa and lpfA), as well as the genetic similarity between them. The occurrence of gene invA was observed in 100% of the samples. The gene lpfA was detected in 80.23% (69/86) strains and is not detected in S. Panama, but present in all strains of S. Infantis. The gene agfA was detected in 63.95% (55/86). S. Agona was positive for all virulence genes studied. The analysis of homology between the different serovars grouped the isolates in clusters. The similarity was regardless of the location of isolation, demonstrating the presence of clones along the production chain and that there are multiple sources for the infection of animals, such as feed, and cross-contamination of carcasses. A survey of virulence genes and evaluation of gene proximity allow characterization and better understanding of Salmonella strains circulating in the pig production chain, thus being able to support control measures during the production process in order to ensure consumer health...

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A diversificação da produção industrial de alimentos de origem suína e o intercâmbio comercial de animais e seus derivados destinados ao consumo humano podem ser importantes disseminadores de sorovares de Salmonella spp. na cadeia alimentar. Objetivou-se avaliar em 86 cepas de Salmonella spp., isoladas em granja de terminação e no abate de suínos, a ocorrência de três genes de virulência (invA, agfA e lpfA), bem como a similaridade genética entre elas. A ocorrência do gene invA foi verificada em 100% das amostras. O gene lpfA foi detectado em 80,23% (69/86) das cepas, não foi detectado em S. Panama e estava presente em todas as cepas de S. Infantis. O gene agfA foi detectado em 63,95% (55/86) das amostras. S. Agona apresentou positividade para todos os genes de virulência estudados. A análise de homologia entre as cepas agrupou os diferentes sorovares em clusters. A similaridade foi independente do local de isolamento, o que demonstra a presença de clones ao longo da cadeia de produção e a existência de multiplicidade de fontes para a infecção dos animais, como a ração, e a contaminação cruzada das carcaças. A pesquisa de genes de virulência e a avaliação da proximidade gênica permitem a caracterização e um maior entendimento sobre cepas de Salmonella circulantes na cadeia produtiva de suínos e, assim, podem subsidiar medidas de controle durante o processo produtivo com o objetivo de garantir a saúde do consumidor.(AU)

The diversification of industrial food production of swine origin and trade of animals and their derivatives for human consumption may be important disseminators of serovars of Salmonella spp. in the food chain. This study aimed to evaluate 86 strains of Salmonella spp. isolated form in the finishing and slaughter of pigs, the occurrence of three virulence genes (invA, agfa and lpfA), as well as the genetic similarity between them. The occurrence of gene invA was observed in 100% of the samples. The gene lpfA was detected in 80.23% (69/86) strains and is not detected in S. Panama, but present in all strains of S. Infantis. The gene agfA was detected in 63.95% (55/86). S. Agona was positive for all virulence genes studied. The analysis of homology between the different serovars grouped the isolates in clusters. The similarity was regardless of the location of isolation, demonstrating the presence of clones along the production chain and that there are multiple sources for the infection of animals, such as feed, and cross-contamination of carcasses. A survey of virulence genes and evaluation of gene proximity allow characterization and better understanding of Salmonella strains circulating in the pig production chain, thus being able to support control measures during the production process in order to ensure consumer health.(AU)

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The objective of this study was to characterize the C. jejuni IAL2383 strain isolated from humans in Brazil. Transcripts for the racR, dnaJ and ciaB genes were found and flaA, plda and cadF genes were present in the genome and bacteria was sensitive to most of the important antimicrobials used to treat humans. C. jejuni IAL2383 is a good experimental model to analyze the interactions with cells.

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The objective of this study was to characterize the C. jejuni IAL2383 strain isolated from humans in Brazil. Transcripts for the racR, dnaJ and ciaB genes were found and flaA, plda and cadF genes were present in the genome and bacteria was sensitive to most of the important antimicrobials used to treat humans. C. jejuni IAL2383 is a good experimental model to analyze the interactions with cells.

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The objective of this study was to characterize the C. jejuni IAL2383 strain isolated from humans in Brazil. Transcripts for the racR, dnaJ and ciaB genes were found and flaA, plda and cadF genes were present in the genome and bacteria was sensitive to most of the important antimicrobials used to treat humans. C. jejuni IAL2383 is a good experimental model to analyze the interactions with cells.(AU)

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Storing hatchable eggs is a common practice in commercial hatcheries. However, storage time may negative effects on several performance parameters. An experiment was carried out to evaluate inverting egg position during storage of eggs laid by young and old layer breeders. Fertile eggs of 32 and 58-week-old breeders were stored for seven, 14, and 21 days at 18ºC ± 2ºC and 80% ± 10% relative humidity (RU). The following parameters were evaluated: egg weight loss, hatchability and hatchling weight, and embryodiagnosis results. Eggs stored with the small end up lost less weight during storage compared with the control eggs. Storing eggs for 14 days with the small end up reduced early embryo mortality, improving hatchability. In addition, hatchling weight increased. These results show that the detrimental effects of long storage periods may be alleviated when eggs are stored with the small end up to 14 days of storage.(AU)

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1. The objective of this study was to evaluate the ability of Campylobacter jejuni to penetrate and colonise eggs from specific-pathogen-free (SPF) and heavy breeder hens, and to determine its effects on the viability of SPF embryos. 2. We detected C. jejuni in 10% of breeder hens and 20% of SPF eggs, which demonstrates the ability of the bacteria to go through the pores of eggs and contaminate the vitellus after 3 h of contact. These results indicate that there is a risk of contamination under commercial production conditions, where, after oviposition, there is contact between the egg and organic material such as faeces and blood. 3. We observed that in 80% of SPF eggs analysed, C. jejuni survived the 21-d incubation period. This positive result suggests that this microorganism was also responsible for early embryonic mortality. 4. The ability of C. jejuni to penetrate the eggs in this study suggests that serious problems may occur under natural field conditions, which may cause significant problems for producers.

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