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Research Highlights: Supplementation with CuNP in feed and water reduced Salmonella Enteritidis count.Supplementation with CuNP did not affect intestinal integrity of broilers.CuNP did not affect weight gain or total lactic acid bacterial counts.The results demonstrate the potential of CuNP as alternative antimicrobials.

1. Salmonella Gallinarum strains isolated from a southern Brazil fowl typhoid outbreak were subjected to phenotypic and genotypic analyses to identify genetic elements that could improve prevention and control strategies.2. Whole-genome sequencing revealed the presence of the aac(6')-Iaa gene, conferring aminoglycoside resistance, along with novel chromosomal point mutations, including the first detection of parE p.S451F in Salmonella Gallinarum.3. Additionally, IncFII(S) plasmid replicons, Salmonella pathogenicity islands and 105 virulence genes associated with cell adhesion, invasion and antimicrobial peptide resistance were identified.4. These findings shed light on the molecular mechanisms of fowl typhoid and provide crucial insights into emerging antimicrobial resistance and virulence factors.

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.

1. Epidemiological surveillance of Salmonella spp. serves as a primary tool for maintaining the health of poultry flocks. Characterising circulating serotypes is crucial for implementing control and prevention measures. This study conducted phenotypic and molecular characterisation of S. enterica Pullorum, S. enterica Heidelberg, and S. enterica Corvalis isolated from broiler chickens during slaughtering.2. All strains were susceptible to gentamicin, neomycin and norfloxacin. However, resistance rates exceeded 50% for ciprofloxacin and tiamulin, irrespective of the serotype. Approximately 64% of strains were classified as multidrug-resistant, with S. enterica Heidelberg strains exhibiting significantly higher overall resistance. The isolates demonstrated the ability to adhere and produce biofilm at a minimum of three temperatures, with S. enterica Pullorum capable of biofilm production at all temperatures encountered during poultry rearing.3. Each strain possessed between two and seven different virulence-associated genes. Genetic similarity, as indicated by pulsed field gel electrophoresis, exceeded 90% for all three serotypes and strains were classified in the R5 ribotype by PCR, regardless of serotype. Sequencing revealed high similarity among all strains, with homology ranging from 99.61 to 100% and all were classified to a single cluster.4. The results suggested a clonal relationship among the strains, indicating the possible circulation of a unique clonal group of S. enterica Pullorum in the southern region of Brazil.

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 (R 2 ), with the highest R 2 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.

Salmonellosis is one of the most common foodborne diseases worldwide. Surface adherence and biofilm formation are among the main strategies evolved by Salmonella to survive under harsh conditions and are risk factors for its spread through the food chain. Owing to the increase in antimicrobial resistance, there is a growing need to develop other methods to control foodborne pathogens, and bacteriophages have been suggested as a potential alternative for this purpose. The aim of this study was to evaluate bacteriophages as a biological control of Salmonella enterica serotypes to inhibit and remove bacterial biofilms. A total of 12 S. enterica isolates were selected for this study, all of which were biofilm producers. Seven bacteriophages were tested, individually and in a cocktail, for their host range and efficiency of plating (EOP). The phage cocktail was evaluated for its antibiofilm effect against the Salmonella biofilms. Phages UPF_BP1, UPF_BP2, UPF_BP3, UPF_BP6, and 10:2 possessed a broad lytic spectrum and could infect all S. enterica strains. Phages 10:2, UPF_BP6, and UPF_BP3 had high EOP in 10, 9, and 9 out of the 12 S. enterica strains, respectively. The cocktail was able to infect all S. enterica strains and had a high EOP in 10 out of 12 S. enterica isolates, presenting a broader host range than any of the tested single phages. A wide variation of inhibition among strains was observed, ranging from 14.72% to 88.53%. Multidrug-resistant and strong biofilm producer strains showed high biofilm inhibition levels by phage cocktail. Our findings demonstrate the ability of the cocktail to prevent biofilm formation and remove formed biofilms of Salmonella . These results indicate that the phage cocktail is a promising candidate to be used as an alternative for the control of Salmonella biofilms through surface conditioning., Competing Interests: Declaration of conflicting interestsThe author(s) declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article.

Poultry products are recognized as the main source of Salmonella and Campylobacter jejuni infections in humans, while avian pathogenic Escherichia coli may have zoonotic potential and can be transmitted from chicken meat to humans. Biofilm formation contributes to their spread through the food chain. This study aimed to compare the adhesion of Salmonella Enteritidis, E. coli, and C. jejuni strains isolated from poultry, food implicated in outbreaks, and poultry slaughterhouses on three surfaces widely used in poultry production (polystyrene, stainless steel, and polyethylene). S. Enteritidis and E. coli adhesion on the three surfaces tested were not significantly different (p > 0.05). Interestingly, the number of C. jejuni cells on stainless steel (4.51-4.67 log 10  CFU/cm. -2 ) was significantly higher (p = 0.0004) than that on polystyrene (3.80-4.25 log 10  CFU/cm. -2 ), but similar (p > 0.05) to that on polyethylene (4.03-4.36 log 10  CFU/cm. -2 ). However, C. jejuni adhesion was significantly lower (p < 0.05) than S. Enteritidis and E. coli adhesion, regardless of the surface evaluated. In addition, scanning electron microscopy analyses have shown an increased irregularity of the stainless steel surface when compared to polyethylene and polystyrene. These irregularities form small spaces ideal for microbial adhesion., Competing Interests: Declaration of competing interest The authors declare that there is no conflict of interest regarding the publication of this article., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

Owing to its antimicrobial activity, electrochemically activated water (ECAW) is a potential alternative to chemical disinfectants for eliminating foodborne pathogens, including Salmonella Heidelberg, from food processing facilities. However, their antibiofilm activity remains unclear. This study aimed to evaluate the antibiofilm activity of ECAW against S. Heidelberg biofilms formed on stainless steel and polyethylene and to determine its corrosive capacity. ECAW (200 ppm) and a broad-spectrum disinfectant (0.2%) were tested for their antibiofilm activity against S. Heidelberg at 25 °C and 37 °C after 10 and 20 min of contact with stainless steel and polyethylene. Potentiostatic polarization tests were performed to compare the corrosive capacity of both compounds. Both compounds were effective in removing S. Heidelberg biofilms. Bacterial counts were significantly lower with ECAW than with disinfectant in polyethylene, regardless the time of contact. The time of contact and the surface significantly influenced the bacterial counts of S. Heidelberg. Temperature was not an important factor affecting the antibiofilm activities of the compounds. ECAW was less corrosive than the disinfectant. ECAW demonstrated a similar or even superior effect in the control of S. Heidelberg biofilms, when compared to disinfectants, reducing bacterial counts by up to 5 log 10 CFU cm -2 . The corrosion of stainless steel with ECAW was similar to that of commercial disinfectants. This technology is a possible alternative for controlling S. Heidelberg in the food production chain., (© 2023. The Author(s) under exclusive licence to Sociedade Brasileira de Microbiologia.)

Escherichia coli is a part of both animal and human commensal microbiota. Avian pathogenic E. coli (APEC) is responsible for colibacillosis in poultry, an economically important disease. However, the close similarities among APEC isolates make it difficult to differentiate between pathogenic and commensal bacteria. The aim of this study was to determine phenotypic and molecular characteristics of APEC isolates and to compare them with their in vivo pathogenicity indices. A total of 198 APEC isolates were evaluated for their biofilm-producing ability and extended-spectrum β-lactamase (ESBL) production phenotypes. In addition, 36 virulence-associated genes were detected, and the isolates were classified into seven phylogenetic groups using polymerase chain reaction. The sources of the isolates were not associated with biofilms, ESBL, genes, or phylogroups. Biofilm and ESBL production were not associated with pathogenicity. Group B2 had the highest pathogenicity index. Groups B2 and E were positively associated with high-pathogenicity isolates and negatively associated with low-pathogenicity isolates. In contrast, groups A and C were positively associated with apathogenic isolates, and group B1 was positively associated with low-pathogenicity isolates. Some virulence-associated genes showed positive or negative associations with specific phylogenetic groups. None of the individual techniques produced results that correlated with the in vivo pathogenicity index. However, the combination of two techniques, namely, detection of virulence-associated genes and the phylogenetic groups, could help the classification of the isolates as pathogenic or commensal., (© 2023. The Author(s) under exclusive licence to Sociedade Brasileira de Microbiologia.)

Owing to concerns about the antimicrobial resistance of agents that can prevent the growth of Listeria monocytogenes in meat, researchers have investigated natural preservatives with antilisterial effects. However, in vivo application of essential oils and plant extracts usually results in reduced antimicrobial activity in meat products when compared to in vitro studies. This study aimed to evaluate the in vivo antimicrobial activity of cinnamon essential oil, pomegranate, and strawberry tree extracts in slices of dry-cured ham and pork loin against L. monocytogenes . Fragments of sterile dry-cured ham were inoculated with 100 μL cinnamon oil 0.5%, pomegranate, or strawberry crude extract. After 10 min, 100 μL of L. monocytogenes serotype 4b (10 4 colony-forming unit [CFU]/mL) was inoculated, and samples were incubated at 7 °C for 7 d to simulate the processing and storage temperature conditions of dry-cured meat products. L. monocytogenes was detected and quantified. Only strawberry extract presented significant differences ( P  < 0.05) from the control; thus, it was selected for the assay with 2% and 4% salt-treated pork loin. The strawberry tree extract significantly ( P  < 0.05) reduced the growth of L. monocytogenes in dry-cured ham. However, it could not reduce L. monocytogenes growth in pork loin, regardless of the salt concentration. This is the first report on the antimicrobial effect of strawberry tree leaf extract against L. monocytogenes in dry-cured ham., Competing Interests: DECLARATION OF CONFLICTING INTERESTSThe author(s) declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article.

Background: Atherosclerotic cardiovascular disease is prevalent among patients with chronic kidney disease (CKD). In this study, we initially aimed to test whether vascular calcification associated with CKD can worsen atherosclerosis. However, a paradoxical finding emerged from attempting to test this hypothesis in a mouse model of adenine-induced CKD., Methods: We combined adenine-induced CKD and diet-induced atherosclerosis in mice with a mutation in the low-density lipoprotein receptor gene. In the first study, mice were co-treated with 0.2% adenine in a western diet for 8 weeks to induce CKD and atherosclerosis simultaneously. In the second study, mice were pre-treated with adenine in a regular diet for 8 weeks, followed by a western diet for another 8 weeks., Results: Co-treatment with adenine and a western diet resulted in a reduction of plasma triglycerides and cholesterol, liver lipid contents, and atherosclerosis in co-treated mice when compared with the western-only group, despite a fully penetrant CKD phenotype developed in response to adenine. In the two-step model, renal tubulointerstitial damage and polyuria persisted after the discontinuation of adenine in the adenine-pre-treated mice. The mice, however, had similar plasma triglycerides, cholesterol, liver lipid contents, and aortic root atherosclerosis after being fed a western diet, irrespective of adenine pre-treatment. Unexpectedly, adenine pre-treated mice consumed twice the calories from the diet as those not pre-treated without showing an increase in body weight., Conclusion: The adenine-induced CKD model does not recapitulate accelerated atherosclerosis, limiting its use in pre-clinical studies. The results indicate that excessive adenine intake impacts lipid metabolism., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Padalkar, Tsivitis, Gelfman, Kasiyanyk, Kaungumpillil, Ma, Gao, Borges, Dhaliwal, Nasruddin, Saji, Gilani, Schram, Singh, Plummer and Savinova.)

Bovines are carriers of Salmonella spp., a relevant foodborne pathogen, acting as contamination sources in slaughterhouses. Calves are prone to infection, and antimicrobial resistance may occur in such bacteria. This study aimed to determine the prevalence and virulence determinants of Salmonella spp. recovered from calves in the Rio Grande do Sul state, Brazil. Eighty-five calves' carcasses were evaluated (leather and veal meat). Thirteen Salmonella spp. isolates (8%) from 11 animals (13%) were obtained only from leather, indicating that contamination occurred before slaughter and that the meat was safe regarding this aspect. The serotypes S. Minnesota, S. Abony, S. Cerro, and S. Gafsa were identified, and all isolates were multidrug-resistant. The isolates had at least 19 virulence-related genes, and the bla OXA-48 resistance gene was detected in three (23%). The data suggest that treating infections caused by these bacteria may be difficult in animals from these farms and can also be an extended human health problem., (© 2022. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

Campylobacteriosis is one of the most common types of bacterial gastroenteritis affecting humans, and poultry is considered a major source of the causative organism, Campylobacter spp. Broilers may arrive contaminated at slaughterhouses, and transport crates could be considered a potential source of contamination. Thus, cleaning and disinfection procedures are crucial to avoid cross-contamination among flocks. Despite its public health importance in Latin American countries, virulence factors of Campylobacter jejuni remain poorly studied in this region. Thus, this study aimed to: 1) determine the occurrence of contaminated crates at a poultry slaughterhouse, 2) compare the contamination before and after the cleaning and disinfection procedures, and 3) detect virulence-associated genes in C. jejuni strains by PCR. Campylobacter spp. were recovered from 8 of the 10 flocks evaluated, and C. jejuni was detected as the main species. There was no significant difference in the Campylobacter detection or quantification between crates at the reception platform and crates after the cleaning/disinfection processes. However, crates after 24 h of natural drying, presented a significant (P < 0.05) lower amount of Campylobacter cells than before the cleaning and disinfection processes. A negative relationship (R 2  = 0.210, P = 0.045) between environmental conditions and Campylobacter quantification was found for transport crates after 24 h of natural drying. There was no significant difference (P > 0.05) in the detection of two C. jejuni virulence genes, flaA (encode a major flagellin protein) and cadF (encode an adhesion and fibronectin-binding protein), among various stages of the cleaning and disinfection processes. Our results demonstrate the high contamination levels of Campylobacter strains in broiler flocks and the potential involvement of poultry transport crates in transmitting these bacteria. This study also suggests that ineffective cleaning and disinfection procedures can increase Campylobacter contamination and facilitate the spread of bacteria in poultry establishments., (Copyright © 2022 The Authors. Published by Elsevier Inc. All rights reserved.)

Biofilm formation has been suggested to play a significant role in the survival of pathogens in food production. Interest in evaluating alternative products of natural origin for disinfectant use has increased. However, there is a lack of information regarding the effects of biosurfactants and organic acids on Salmonella enterica serotype Enteritidis, Escherichia coli, and Campylobacter jejuni biofilms, mainly considering temperatures found in environments of poultry processing, as well as simulating the contact times used for disinfection. The aim of this study was to evaluate the antibiofilm activity of rhamnolipid, malic acid, and citric acid on the adhesion of S. Enteritidis, E. coli, and C. jejuni on polystyrene surfaces at different temperatures (4, 12, and 25 °C), compound concentrations, and times of contact (5 and 10 min), and to analyze the potential use of these compounds to disrupt formed biofilms. All three compounds exhibited antibiofilm activity under all analyzed conditions, both in the prevention and removal of formed biofilms. Contact time was less important than temperature and concentration. The antibiofilm activity of the compounds also varied according to the pathogens involved. In the food industry, compound selection must consider the temperature found in each stage of product processing and the target pathogens to be controlled., (© 2022. The Author(s) under exclusive licence to Sociedade Brasileira de Microbiologia.)

Background: Listeria monocytogenes is a widespread common contaminant in food production facilities during preparation, storage, and distribution, and minimally processed ready-to-eat products are considered at high risk of contamination by this bacterium. Increased antibiotic resistance has led researchers to search for plant-based natural alternatives to control pathogenic microorganisms. Among these products, essential oils and plant extracts have previously shown antimicrobial activity and are possible alternatives to manage food pathogens. In this study, commercial essential oils (cinnamon, clove, oregano, ginger, and thyme) and plant extracts (pomegranate, acorn, olive, strawberry tree, and dog rose) were tested against L. monocytogenes in a dry-cured ham-based model., Results: Essential oils and plant extracts were screened by agar diffusion and minimum inhibitory concentration for anti-L. monocytogenes activity. Cinnamon, pomegranate, and strawberry trees returned the strongest results and were therefore evaluated in a dry-cured ham-based medium assay with water activity of 0.93 or 0.95. The 10% essential oil of cinnamon was capable of completely inhibiting bacterial growth, while strawberry tree and pomegranate extract also showed antilisterial activity (P > 0.05). Water activity influenced the bacterial count of L. monocytogenes in a dry-cured ham-based medium., Conclusions: There was a reduction in L. monocytogenes with the application of cinnamon essential oil but, because of the negative sensory impact of this particular compound in meat products, we suggest the use of pomegranate or strawberry tree for the biocontrol of Listeria in ready-to-eat products. © 2021 Society of Chemical Industry., (© 2021 Society of Chemical Industry.)

Poultry products are susceptible to contamination by pathogenic and spoilage bacteria during the slaughtering process. Molecular techniques have been used to assist in the identification of microorganisms in various microbiomes. The aim of this study was to identify bacterial components of the microbiome in poultry carcasses during the slaughter process, using high-throughput next generation sequencing (HT-NGS). Samples were collected from three slaughterhouses (A, B, and C) located in southern Brazil and included those taken from three points (initial, middle, and end) in the chiller tanks and two carcass pools (at the entrance to the clean area and after the final carcass packaging) at each establishment. A total of 104 carcasses were collected from each slaughterhouse. For this study, HT-NGS allows for a precise, quantitative and culture-independent microbiome assessment in poultry products. Three phyla (Firmicutes, Bacteroidetes, and Proteobacteria) were found in all establishments, and one phylum (Verrucomicrobia) was found only in Establishment A. Common set of genera (Anaerotruncus, Bacteroides, Campylobacter, Erysipelatoclostridium, Faecalibacterium, Lachnoclostridium, and Subdoligranulum) was identified in processing establishments along with the groups unique to a particular site. Pathogenic and spoilage bacteria, as well as other microorganisms that were not expected in poultry products, were detected by HT-NGS technique. The Shannon diversity index was the highest in Establishment B (2.40), followed by establishments C (1.98) and A (1.43). As we progressed through sample analysis, from the entrance of the clean area to the final carcass packaging area, we found significant reductions (p < 0.05) in the quantities of sequences of all phyla in establishments A and B. Significant differences (p < 0.05) in the quantities of sequences of all phyla were found between different stages in the slaughtering process. More stringent control procedures in establishments A and B were associated with reduced contamination even though all establishments followed the official sanitary standards. Our findings provide new insight into the chicken meat microbiome, and can be used in future studies to help ensure food safety in slaughterhouses., (Copyright © 2022 Elsevier B.V. All rights reserved.)

Competing Interests: We declare no competing interests. PVJ, AG, and KAB contributed equally to this Comment. The Yale University institutional review board approved the study (2000027792) and waived the need for informed consent.

As a consequence of developing antimicrobial resistance to disinfectants, copper, which exhibits antimicrobial activity, has been studied as a possible alternative to the use of stainless steel surfaces. The aim was to evaluate the antimicrobial activity of copper surfaces in preventing biofilm formation by Salmonella Enteritidis and to determine their corrosive capacity. Strains of S. Enteritidis were incubated at 4 °C, 12 °C, and 25 °C with 1 cm 2 coupons of electrolytic copper (99.9% Cu), brass (70% Cu), copper coated with tin, and stainless steel (control). A planktonic cell-suspension assay was used, followed by serial dilutions and bacterial counts. The corrosion test was performed with two disinfectants: benzalkonium chloride and sodium hypochlorite (100, 200, and 400 ppm). There was a significant reduction in biofilm production (log 10  CFU cm -2 ) on the copper (2.64 at 4 °C, 4.20 at 12 °C, 4.56 at 25 °C) and brass (2.79 at 4 °C, 3.49 at 12 °C, 4.55 at 25 °C) surfaces compared to the control (5.68 at 4 °C, 5.89 at 12 °C, 6.01 at 25 °C). The antimicrobial surfaces showed uniform corrosion similar to that of surfaces generally used. These results demonstrated the effectiveness of copper surfaces in reducing S. Enteritidis and suggest they can be used as a complementary antimicrobial to control for this pathogen., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

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&#95;BP1 and UPF&#95;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&#95;BP1 and UPF&#95;BP2 were shown to be promising alternatives for the biological control of fowl typhoid.

Brazil is the number one exporter of chicken meat, and this industry maintains constant microbiological vigilance. The objective of this study was to characterize the pathogenicity, antimicrobial resistance (AMR) and the profile of biofilm production of Escherchia coli strains isolated from raw refrigerated cuts of chicken meat sold in retail markets of the four largest poultry companies in Brazil. We collected 150 samples of chicken meat, in order to isolate E. coli and performed susceptibility tests (to amoxicillin associated with clavulanic acid, ceftiofur, enrofloxacin, gentamicin, and trimethoprim + sulfamethoxazole). In addition, the disc approximation test to detect extended spectrum beta-lactamases enzymes (ESBLs) producers was performed. E. coli ability to form biofilm was checked using polystyrene microplates. We also searched for ESBLs genes (blaCTY-M2, blaSHV-1, blaTEM-1, blaCTX-M2, blaOXA-1, blaPSE-1 and AmpC) and adhesion genes (sfa/foc, afa/draB, iha, hrla, fimC, tsh, papC, mat, cr1, felA, fimH and papG) in ESBL-E. coli producers and in those E. coli classified as strongly biofilm formers, respectively. The overall percentage of E. coli isolation was 58.66%, with brand A having the highest percentage (70%), followed by brands D, B and C (60, 53.3 and 50%, respectively). The highest resistance profile was observed for beta-lactams (39.5%), followed by sulfonamide associated to trimethoprim (36.9%) and polymyxin (33.4%). Of the isolates obtained, 77% were non-susceptible to at least one antimicrobial. Brand A showed the highest overall percentage of resistance with 95.23%, followed by brands C (80%), B (75%) and D (69.44%). Overall, 73.86% of the isolates were non susceptible to at least one antibiotic and 36.3% were multiresistants. A total of 17.04% of E. coli strains were identified as ESBLs producers and 70.44% were able to form biofilms (moderate-to-strong). The blaTEM-1 gene was the most prevalent (73.33%), followed by blaSHV-1 (46.66%) and blaCMY-2 (6%). Of the 31 strongly biofilm-forming strains, 26 (83.87%), 24 (77.41%) and 20 (64.51%) expressed fimC, papG and crl genes, respectively. Taken together, our results show that Brazilian chicken meat can be contaminated with E. coli that are non-susceptible to multiple antibiotics, able to form biofilm and showing a diverse repertoire of adhesins linked to pathogenicity depending on the brand evaluated., Competing Interests: Declaration of Competing Interest The authors declare that they have no conflict of interest., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Overpopulation of domestic pigeons is considered to be one of the major problems of urban centers, as these birds are responsible for the dissemination of relevant pathogens to animal and human health. The aim of this study was to detect potentially pathogenic Escherichia coli and Salmonella spp. in domestic pigeons captured in areas near silos used for grain and feed storage, analyzing the antimicrobial sensitivity and the presence of virulence-associated genes. We evaluated 41 pigeons. From each bird, cecal contents and a pool of viscera (heart, spleen, and liver) were collected. Fifty strains of E. coli and three strains of S. Typhimurium were isolated. The antimicrobial susceptibility assay showed that 2% of the isolates of E. coli were resistant to chloramphenicol and the combination of sulfamethoxazole + trimethoprim and 4% to tetracycline, doxycycline, and sulfonamide. The three S. Typhimurium strains were sensitive to all antimicrobials tested. The pathogenicity profile demonstrated that no E. coli isolates showed a STEC compatible profile. Regarding the APEC pathotype, all genes were observed in 8% of E. coli, 6% had only the iss gene and 4% presented ompT, hlyF, and iutA genes. invA, hilA, avrA, and lpfA genes were detected in 100% of Salmonella isolates. The sitC and pefA genes were only present in one strain and the remaining genes were detected in two. In conclusion, it was found that pigeons living in the vicinity of silos are carriers of important pathogens, and control measures should be taken to minimize animal and human health risks.

Salmonella remains one of the most common foodborne pathogens worldwide, and its resistance to antimicrobials and disinfectants has increased considerably over the years. Thus, monitoring its resistance to products commonly used in swine production is indispensable for the development of strategies to reduce the occurrence of bacterial resistance. In this context, our aim was to detect Salmonella at different points in swine slaughterhouses, identify the main serotypes, and evaluate their resistance to disinfectants and antimicrobials used in swine production. Salmonella at the processing plants was detected by conventional microbiology. Salmonella strains were tested for susceptibility to peracetic acid (0.5% and 1%), quaternary ammonium (0.5%), and seven antimicrobials. Twenty-eight percent of the samples were positive for Salmonella, with the most identified serotypes being Salmonella Derby, Salmonella Typhimurium, and monophasic Salmonella Typhimurium. All tested strains were susceptible to both concentrations of peracetic acid, but only 28% were susceptible to quaternary ammonium. Sixteen percent of the strains were susceptible to all tested antimicrobials. Only enrofloxacin was efficient in inhibiting the growth of all strains. The highest number of non-susceptible strains was to amoxicillin, followed by chloramphenicol, florfenicol, and doxycycline. Thirty-six percent of the strains were classified as multidrug-resistant. Salmonella were detected in all slaughtering processes, and important serotypes were recovered, including Salmonella Typhimurium, Salmonella Derby, monophasic Salmonella Typhimurium, and Salmonella Infantis. We observed high rates of resistance to quaternary ammonium and to important antimicrobial agents. Salmonella Typhimurium and Salmonella Infantis were the most resistant serotypes.

Introduction: Campylobacteriosis is considered the most common bacteria-caused human gastroenteritis in the world. Poultry is a major reservoir of Campylobacter. Human infection may occur by consumption of raw and undercooked poultry or by contamination of other foods by these items. The aim of this study was to assess the prevalence of Campylobacter spp. in poultry processing plants with conventional culture method and real-time PCR., Methodology: A total of 108 poultry processing plant samples were collected to test with conventional microbiology and qPCR. Sampling included cloacal swabs, swabs of transport crates (before and after the cleaning and disinfection process) and carcasses (after the chiller, cooled at 4°C and frozen at -12°C)., Results: Positivity in cloacal swabs indicated that poultry arrived contaminated at the slaughterhouse. Contamination in transport cages was substantially increased after the cleaning process, indicating that the process was ineffective. The detection of Campylobacter on carcasses was higher than that on cloacal swabs, which could indicate cross-contamination during the slaughtering process. Conventional microbiology and molecular methods revealed a prevalence of 69.4% and 43.5%, respectively. Lower detection by qPCR can be attributed to the high specificity of the kit and to biological components that could inhibit PCR reactions., Conclusions: Our results indicate that poultry arrive contaminated at the slaughterhouse and that contamination can increase during the slaughtering process due to cross-contamination. The isolation of Campylobacter in cooled and frozen carcasses corroborates the bacterial survival even at temperatures considered limiting to bacterial growth which are routinely used for food preservation., Competing Interests: No Conflict of Interest is declared, (Copyright (c) 2020 Karen Apellanis Borges, Isabel Cristina Cisco, Thales Quedi Furian, Denise Cristina Tedesco, Raíssa Canova, Laura Beatriz Rodrigues, Vladimir Pinheiro do Nascimento, Luciana Ruschel dos Santos.)

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.

Introduction: Salmonella is a major cause of foodborne illness throughout the world. The use of quantitative techniques is important for assessing the risk and determining the capacity of each step of the slaughtering process to decrease or increase bacterial contamination. We aimed to detect and to quantify the presence of Salmonella in Brazilian processing plants by real-time quantitative polymerase chain reaction (qPCR)., Methodology: A total of 139 poultry slaughterhouses samples were collected in order to detect to and quantify Salmonella by qPCR., Results: Almost all collection points (3/18), except water from pre-chiller tank, carcasses after pre-chiller, and carcasses frozen at -12ºC for 60 days, and 49% (68/139) of samples were positive for Salmonella. Quantification means varied equally among all of the tested sources, and we could not establish any pattern of variation. A large proportion (52.6%) of cloacal swabs was Salmonella-positive. Also, contamination in transport cages was increased after the cleaning process, indicating that the process was ineffective. The overall prevalence in samples obtained during the slaughtering process was 48.9%, and on the whole rinsed carcasses, this proportion was 50%. The detection of Salmonella in frozen carcasses, even after long periods of storage, indicates that the carcasses are a potential source of infection for consumers., Conclusions: We found that contamination levels remain similar throughout the slaughtering. qPCR proved to be an efficient method for the detection of Salmonella., Competing Interests: No Conflict of Interest is declared, (Copyright (c) 2019 Karen Apellanis Borges, Eduarda Boff Martelo, Lilian Andriva dos Santos, Thales Quedi Furian, Isabel Cristina Cisco, Luciane Manto, Luciana Ruschel dos Santos.)

Salmonella spp. are the main pathogens responsible for foodborne disease worldwide. Bacterial communities use the quorum sensing system to control biofilm formation. These systems function through the secretion of substances, called auto-inducers (AI), into the environment. AI-3 is structurally similar to epinephrine (EPI) and norepinephrine (NOR) -catecholamines secreted by eukaryotic cells to communicate with each other. In this context, this work aimed to evaluate the effect of EPI and NOR on biofilm formation by S. Enteritidis at 12 °C and 25 °C. Also, we detected the presence of the csgD, adrA, and fimA genes in these strains. Biofilm formation was investigated at two temperatures (12 °C and 25 °C) using a microtiter plate assay, under four different treatments (50 mM EPI, 100 mM EPI, 50 mM NOR; 100 mM NOR) and a control group. PCR was used to detect the virulence genes associated with biofilm production. A greater number of biofilm producer isolates were observed at 25 °C than at 12 °C, regardless of the treatment. The number of biofilms forming strains at 12 °C was significantly higher in the treatment with norepinephrine at 100 μM. The proportion of non-producer and biofilm producer strains at 25 °C did not differ significantly among the treatments. All strains presented the three genes (csgD, adrA, and fimA). The approach carried out in this work is a precursor in veterinary medicine, focusing on both public and poultry health, and evaluates the influence of catecholamines on the formation of biofilms with S. Enteritidis, an important pathogen with zoonotic potential. Norepinephrine seems to be more efficient at stimulating biofilm formation by S. Enteritidis strains at 12 °C. csgD, fimA, and adrA were detected in all strains., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Avian pathogenic Escherichia coli (APEC) is one of the pathogens that most concerns the poultry industry worldwide due to the economic losses it can cause. APEC persistence and survival, both in the environment and in the host, may be a consequence of biofilm-producing capabilities. The aim of this study was to evaluate APEC strains' biofilm production and its relationship to in vivo pathogenicity. Two hundred thirty-eight APEC isolates from three different origins (broiler bedding material, cellulite lesions, and respiratory diseases) were selected. The in vivo pathogenicity index (PI) was determined. Biofilm formation was evaluated using a microplate assay with analysis of colony morphology in Congo Red agar in order to detect the phenotypic expression of curli fimbriae and cellulose. Regarding biofilm production, it was observed that 55.8% of the strains produced biofilms. In the morphological test, 88.2% of the isolates expressed one or both components at one of the temperatures at least, and 11.8% of the isolates did not express curli or cellulose. Cellulose production was significantly higher at 25 °C. On the other hand, curli production was significantly higher at 37 °C. The study data indicate that there is no association between biofilm production and in vivo pathogenicity.

Background: Hepatocellular carcinoma (HCC) is a common malignancy with a steadily rising incidence and associated morbidity and mortality. Cirrhosis of the liver is presently the leading risk factor for developing HCC. Abdominal imaging, with or without alpha-fetoprotein (AFP) testing, every 6 months is the current surveillance strategy for patients at risk. The available biomarkers for detecting this cancer at an early stage have inadequate sensitivity and specificity., Methods: The Hepatocellular carcinoma Early Detection Strategy (HEDS) study, a multi-center initiative of the National Cancer Institutes' (NCI) Early Detection Research Network (EDRN), launched an effort to establish what has become the nation's largest comprehensive biorepository and database on patients at high risk of developing HCC. The cohort has been developed in seven clinical centers across the USA. Subjects are enrolled for a five-year period involving data and specimen collection every six months in accordance with standard surveillance for HCC. Extensive clinical data are collected and specimens are stored at a central repository., Results: The database and biorepository contain longitudinally collected clinical data and serum and plasma samples from 1482 participants with cirrhosis and without evidence of HCC at baseline. Fifty-six percent are male, 85% Caucasian, 30% have a history of chronic HCV and 71% have compensated cirrhosis., Conclusions: The HEDS cohort provides opportunities for the continued study of the incidence and course of HCC in a comprehensively followed population of patients at high risk for this malignancy. Further, the EDRN biorepository provides a distinct opportunity for the development of novel biomarkers. Trial registry URL: https://edrn.nci.nih.gov/protocols/316-hepatocellular-carcinoma-early-detection-strategy., (Copyright © 2018 Elsevier Inc. All rights reserved.)

Fowlpox is one of the oldest diseases reported in birds. The causative genus Avipoxvirus affects ~232 domestic and wild species. We present herein the history, clinical findings, and macroscopic and histologic lesions caused by a clade C poxvirus in an exotic psittacine breeding colony in southern Brazil. Clinical signs included yellow nodular lesions at the commissure of the beak and on the periocular skin, loss of appetite, and death. Fifty birds were autopsied, and fragments of periocular skin, tongue, and trachea were examined histologically, which revealed hyperkeratosis associated with eosinophilic intracytoplasmic inclusion bodies. Tracheal fragments and periocular skin were subjected to nested PCR and phylogenetic analyses. The sequenced strain showed 99.58% identity with the nucleotide sequences of Avipoxvirus strains AY53011, KC018069, AM050383, and AM05382 isolated from birds in Germany, United States, and United Kingdom. The strain was grouped under clade C, which represents isolates exclusively from the Psittacidae family. The infection caused by clade C Avipoxvirus in the exotic psittacines examined ( Platycercus sp. and Psephotus haematonotus) demonstrates the circulation of this clade in this breeding colony.

Salmonella Enteritidis and Salmonella Typhimurium are among the most prevalent serotypes isolated from salmonellosis outbreaks and poultry. Salmonella spp. have the capacity to form biofilms on several surfaces, which can favour survival in hostile environments, such as slaughterhouses. Salmonella strains present differences in pathogenicity. However, there is little information regarding the pathogenicity of S. Enteritidis and S. Typhimurium isolated from avian sources and their relationship to biofilm production. The aim of this study was to use a novel pathogenicity index and a biofilm production assay to evaluate their relationships within these serotypes. In addition, we detected the presence of the spiA and agfA genes in these strains. Biofilm formation was investigated at two temperatures (37 °C and 28 °C) using microtiter plate assay, and the results were compared with the individual pathogenicity index of each strain. PCR was used to detect spiA and agfA, virulence genes associated with biofilm production. S. Enteritidis and S. Typhimurium strains were capable of producing biofilm at 37 °C and 28 °C. Sixty-two percent and 59.5% of S. Enteritidis and 73.8% and 46.2% of S. Typhimurium produced biofilm at 37 °C and 28 °C, respectively. Biofilm production at 37 °C was significantly higher in both serotypes. Only S. Enteritidis was capable of adhering strongly at both temperatures. Biofilm production was related to pathogenicity index only at 28 °C for S. Enteritidis. spiA and agfA were found in almost all strains and were not statistically associated with biofilm production., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

Salmonella Enteritidis remains a standout among the leading causes of foodborne diseases worldwide. Previous studies have demonstrated that a unique clonal group of Salmonella Enteritidis, named SE86, is involved in foodborne outbreaks in southern Brazil and is frequently identified among strains isolated from poultry. The aim of this study was to determine the influence of the isolation source (food products involved in salmonellosis outbreaks and poultry sources) on the phenotypic and molecular characteristics of Salmonella Enteritidis SE86. A biofilm formation assay, antimicrobial susceptibility test, polymerase chain reaction identification of virulence-associated genes, and phage type 4 (PT4) assessment were performed to characterize Salmonella Enteritidis SE86. The human strains presented less antimicrobial resistance than the poultry strains. Resistance to some substances was related to the isolation source of the strain. Strains of the same clonal group presented different biofilm production abilities. Biofilm formation was independent of the isolation source at all temperatures. Temperature influenced biofilm formation only by the poultry strains. Most of the investigated genes presented a high frequency and a regular distribution, regardless of the isolation source. The spvB, spiA, pagC, sipB, prgH, spaN, sitC, and lpfC genes were associated with the avian strains, whereas iroN was associated with the strains isolated from food products involved in salmonellosis outbreaks. Most strains belonged to PT4. No relationship was found between biofilm production and antimicrobial resistance or between the virulence profile and biofilm production or antimicrobial resistance.

Rational and Objectives: Low intensity vibration (LIV) may represent a nondrug strategy to mitigate bone deficits in patients with end-stage renal disease., Materials and Methods: Thirty end-stage renal patients on maintenance hemodialysis were randomized to stand for 20 minutes each day on either an active or placebo LIV device. Analysis at baseline and completion of 6-month intervention included magnetic resonance imaging (tibia and fibula stiffness; trabecular thickness, number, separation, bone volume fraction, plate-to-rod ratio; and cortical bone porosity), dual-energy X-ray absorptiometry (hip and spine bone mineral density [BMD]), and peripheral quantitative computed tomography (tibia trabecular and cortical BMD; calf muscle cross-sectional area)., Results: Intention-to-treat analysis did not show any significant changes in outcomes associated with LIV. Subjects using the active device and with greater than the median adherence (70%) demonstrated an increase in distal tibia stiffness (5.3%), trabecular number (1.7%), BMD (2.3%), and plate-to-rod ratio (6.5%), and a decrease in trabecular separation (-1.8%). Changes in calf muscle cross-sectional area were associated with changes in distal tibia stiffness (R = 0.85), trabecular bone volume/total volume (R = 0.91), number (R = 0.92), and separation (R = -0.94) in the active group but not in the placebo group. Baseline parathyroid hormone levels were positively associated with increased cortical bone porosity over the 6-month study period in the placebo group (R = 0.55) but not in the active group (R = 0.01). No changes were observed in the nondistal tibia locations for either group except a decrease in hip BMD in the placebo group (-1.7%)., Conclusion: Outcomes and adherence thresholds identified from this pilot study could guide future longitudinal studies involving vibration therapy., (Copyright © 2017 The Association of University Radiologists. Published by Elsevier Inc. All rights reserved.)

Salmonella spp. are among the most important agents of foodborne diseases all over the world. Human Salmonella outbreaks are often associated with the consumption of poultry products (meat and eggs), and one of the most prevalent serotypes associated with these products is Salmonella Enteritidis. Brazil is one of the most important poultry exporters in the world. In southern Brazil, three closely related clones of Salmonella Enteritidis have been responsible for the majority of foodborne Salmonella outbreaks over the past decade. However, until now, there has been little information regarding the clonal relationship among the Brazilian Salmonella strains of avian origin and those involved in foodborne outbreaks. Therefore, the aim of the present study was to complete the molecular characterization of Salmonella Enteritidis strains isolated from poultry and food sources involved in Salmonella outbreaks. PCR ribotyping was performed to discriminate the strains into different ribotype profiles according to the banding pattern amplification. This technique was able to differentiate the Salmonella Enteritidis strains into two banding patterns: R2 and R4. R2 accounted for 98.7% of the strains. DNA sequencing of the 600-bp fragment, present in all ribotypes, was applied to confirm this result. The sequences generated showed high levels of similarity, ranging from 99.7 to 100%, and were grouped into a single cluster. These results suggest that there is a clonal relationship among the Salmonella Enteritidis strains responsible for several salmonellosis outbreaks and the strains collected from poultry sources.

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 .

Pasteurella multocida causes atrophic rhinitis in swine and fowl cholera in birds, and is a secondary agent in respiratory syndromes. Pathogenesis and virulence factors involved are still poorly understood. The aim of this study was to detect 22 virulence-associated genes by PCR, including capsular serogroups A, B and D genes and to evaluate the antimicrobial susceptibility of P. multocida strains from poultry and swine. ompH, oma87, plpB, psl, exbD-tonB, fur, hgbA, nanB, sodA, sodC, ptfA were detected in more than 90% of the strains of both hosts. 91% and 92% of avian and swine strains, respectively, were classified in serogroup A. toxA and hsf-1 showed a significant association to serogroup D; pmHAS and pfhA to serogroup A. Gentamicin and amoxicillin were the most effective drugs with susceptibility higher than 97%; however, 76.79% of poultry strains and 85% of swine strains were resistant to sulphonamides. Furthermore, 19.64% and 36.58% of avian and swine strains, respectively, were multi-resistant. Virulence genes studied were not specific to a host and may be the result of horizontal transmission throughout evolution. High multidrug resistance demonstrates the need for responsible use of antimicrobials in animals intended for human consumption, in addition to antimicrobial susceptibility testing to P. multocida., (Copyright © 2015 Sociedade Brasileira de Microbiologia. Published by Elsevier Editora Ltda. All rights reserved.)

Background: The most common hereditary hemoglobin disorder, affecting 20 million individuals worldwide, is sickle cell disease. The vascular obstruction resulting from the sickling of cells in this disease can produce local hypoxemia, pain crises and infarction in several tissues, including the bones, spleen, kidneys and lungs., Objective: To determine red blood group genes in a Brazilian populations., Methods: The present study is characterized as a case control study, with the aim of identifying the baseline blood lactate concentration in individuals with hemoglobin SS and SC diseases. One-way ANOVA with the Tukey post-test was used to analyze the results and a p-value < 0.05 was considered significant. Calculations were made using the INSTAT statistical program. The graphs were generated using the ORING program. The study sample was composed of 31 men and women residing in the city of Santo Antônio de Jesus, Bahia, Brazil. The individuals were divided into two groups: Group GC of 16 subjects who did not present with any type of structural hemoglobinopathy; and Group GE composed of 15 individuals with ages between 2 and 35 years old, who had the SS and SC genotypes. Sample analyses were performed with 3 mL of blood during fasting., Results: The baseline blood lactate concentration of the SS and SC individuals was higher than that of the control group (p<0.001) with means of 4.86 ± 0.95; 3.30 ± 0.33; 1.31 ± 0.08 IU/L for SS, SC and controls, respectively. This corroborates the initial research hypothesis., Conclusion: The baseline blood lactate of SS and SC individuals is 3 to 4 times higher than that of healthy subjects, probably due to the fact that these patients have a metabolic deviation to the anaerobic pathway.

Rabbit proactivator is a neutral metalloproteinase that activates another metalloproteinase, procollagenase, and degrades noncollagenous matrix. We describe the construction of an activator complementary DNA (cDNA) clone, which is 1.9 kb, that selects a 2.1-kb messenger RNA (mRNA) in Northern blot hybridizations. Nucleic acid sequence studies of the activator cDNA indicate 1) that it encodes protein Mr 53,881, 2) that this protein exhibits approximately 80% homology with rat transin, an oncogene-induced protein with a previously unknown function, and 3) that, in the first 172 residues, it is virtually identical to the rabbit metalloproteinase, stromelysin. Homology between rabbit activator and human skin collagenase is approximately 50%. Activator and collagenase mRNA are coordinately regulated; untreated cultures of rabbit synovial fibroblasts produce low levels of each protein, but addition of phorbol myristate acetate (10(-8)M) results in an increase in mRNA for both proteins by 2.5-5 hours. Adding all-trans-retinoic acid (10(-6)M) or dexamethasone (10(-7)M) to phorbol-stimulated cells coordinately suppresses both activator and collagenase mRNA. Our data suggest the existence of coordinately regulated metalloproteinases that are important in the modulation of connective tissue metabolism.