Antibiofilm activity of electrochemically activated water (ECAW) in the control of Salmonella Heidelberg biofilms on industrial surfaces.

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 log10 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.

Antimicrobial and antibiofilm activity of silver nanoparticles against Salmonella Enteritidis.

Salmonella enterica serotype Enteritidis is one of the main pathogens associated with foodborne illnesses worldwide. Biofilm formation plays a significant role in the persistence of pathogens in food production environments. Owing to an increase in antimicrobial resistance, there is a growing need to identify alternative methods to control pathogenic microorganisms in poultry environments. Thus, this study aimed to synthesize silver nanoparticles (AgNPs) and evaluate their antibiofilm activity against poultry-origin Salmonella Enteritidis in comparison to a chemical disinfectant. AgNPs were synthesized, characterized, and tested for their minimum inhibitory concentration, minimum bactericidal concentration, and antibiofilm activity against S. Enteritidis strains on polyethylene surfaces. The synthesized AgNPs, dispersed in a liquid medium, were spherical in shape with a mean diameter of 6.2 nm. AgNPs exhibited concentration-dependent bactericidal action. The bacterial reduction was significantly higher with AgNPs (3.91 log10 CFU [Formula: see text] cm-2) than that with sanitizer (2.57 log10 CFU âˆ™ cm-2). Regarding the time of contact, the bacterial count after a contact time of 30 min was significantly lower than that after 10 min. The AgNPs exhibited antimicrobial and antibiofilm activity for the removal of biofilms produced by S. Enteritidis, demonstrating its potential as an alternative antimicrobial agent. The bactericidal mechanisms of AgNPs are complex; hence, the risk of bacterial resistance is minimal, making nanoparticles a potential alternative for microbial control in the poultry chain.

Bacteriophages as an alternative for biological control of biofilm-forming Salmonella enterica.

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.

Prevalence and distribution of pathogenic genes in Campylobacter jejuni isolated from poultry and human sources.

INTRODUCTION: Campylobacter jejuni is one of the most common bacterial causes of human gastroenteritis. Despite its public health importance, the virulence factors and mechanisms underlying C. jejuni pathogenesis remain poorly understood and the relationships between these genes and the sources of the strains are not clear. We aimed to determine the virulence profiles of C. jejuni isolated from poultry and human cases of Campylobacteriosis. METHODOLOGY: A total of 50 strains of C. jejuni isolated from poultry and human cases of Campylobacteriosis were screened by polymerase chain reaction (PCR) for the presence of six pathogenic genes (flaA, iam, wlaN, cdtA, cdtB, cdtC). RESULTS: A total of 40% (10/25) of the human isolates presented only one virulence marker. In contrast, 64% (16/25) of the poultry-derived strains showed four or five virulence markers. cdtA and flaA occurred more frequently in poultry-derived strains than in human strains. Ten different virulence profiles were observed among the human isolates and 11 among the poultry strains. Only four profiles were common to both sources: profiles 3 (flaA, cdtA, cdtB, and cdtC), 5 (cdtA and cdtB), 7 (flaA and cdtB), and 10 (iam, flaA, cdtA, cdtB, and cdtC). The human-derived strains had a higher Shannon diversity index (1.9396) and Simpson index (0.8367), indicating a more diversified population than found in poultry (1.7742 and 0.7333, respectively). CONCLUSIONS: We found variations in the genetic profiles of the circulating strains based on the isolation source and genes that are potentially pathogenic to humans were detected in poultry-derived strains.

Detection and Quantification of Campylobacter in Poultry Slaughterhouses Using Conventional Microbiological Technique, Most Probable Number, and Real-Time PCR.

Campylobacteriosis is one of the most common bacteria causing human gastroenteritis. Poultry is a major reservoir of Campylobacter spp. as well as the main source of transmission. Due to the increased occurrence of campylobacteriosis, poultry slaughterhouses are under pressure to deliver carcasses with low contamination. However, a few studies have been carried out to evaluate Campylobacter contamination of broiler carcasses in Brazilian slaughter lines. Therefore, in this study, we aimed at detecting and quantifying the thermotolerant Campylobacter spp. at different stages of the poultry slaughtering process. The samples were collected from 12 points in three slaughterhouses in southern Brazil, at an interval of 12 months, and were tested for Campylobacter spp. by conventional microbiological technique, the most probable number, and real-time PCR. A total of 432 samples were analyzed. The majority of strains belonged to Campylobacter jejuni (92%), and the flock positivity among the three techniques was similar in most cases. Campylobacter was detected in all slaughtering stages. Although contamination has remained similar (p > 0.05) throughout almost all the slaughter process, evisceration seemed to be an important source of contamination. Our results reinforce the idea that the final carcass quality after the slaughtering process is directly influenced by the level of contamination of the broiler flocks on arrival at the processing plant.

Identification of pathogenic genes in Campylobacter jejuni isolated from broiler carcasses and broiler slaughterhouses.

Campylobacter jejuni is one of the most common causes of foodborne diseases worldwide. There are few reports on Campylobacter strains isolated from Latin-American countries. Here, 140 C. jejuni strains isolated from cloacal and transport boxes swabs, water from chiller tanks, and broiler carcasses of five poultry companies in Southern Brazil were identified using phenotypic and genotypic methods. Polymerase chain reaction (PCR) was used to analyze eight C. jejuni virulence markers: flaA, cadF, and invasion-associated (iam) genes, cdtABC operon (associated with the cytolethal distending toxin), and plasmidial virB11 and wlaN genes were present in 78.5%, 77.8%, 0%, 74.2%, 22.1%, and 10.7% of samples, respectively. There were 25 different virulence profiles: 1 (cdtA, cdtB, cdtC, flaA, and cadF), 2 (cdtA, cdtB, cdtC, flaA, cadF, and virB11), and 3 (cdtA, cdtB, cdtC, flaA, cadF, and wlaN) were the most common (> 60% of strains). We provide insight into factors related to the occurrence of this pathogen and their epidemiology.

Influence of the norepinephrine and medium acidification in the growth and adhesion of Salmonella Heidelberg isolated from poultry.

Salmonella spp. are among the leading pathogens responsible for foodborne illnesses worldwide. Bacterial communities use a quorum sensing (QS) system to control biofilm formation. QS is a cell-to-cell signaling mechanism involving compounds called auto-inducers (AI). Norepinephrine utilizes the same bacterial signaling of AI-3 and serves as a signal of QS. Acid stress is a challenge encountered by microorganisms in food processing environments and in the gastrointestinal tracts of hosts. Thus, adaptation to acidic environments may increase the pathogenicity of the strain. The aim of this study was to evaluate the influence of two concentrations of norepinephrine (100 µM and 250 µM) and acidification (pH 3.0) of the medium on the growth and adhesion of Salmonella Heidelberg strains isolated from poultry sources at 12 °C and 25 °C. Furthermore, three genes associated with the biofilm formation process were detected (adrA, csgD, and sidA). Norepinephrine stimulation did not influence the growth or adhesion of Salmonella Heidelberg strains, regardless of the catecholamine concentration and temperature. On the other hand, the use of acidified medium (pH 3.0) resulted in a significant reduction of growth and a significant increase of S. Heidelberg adhesion at both temperatures, indicating that the acidified medium favors the biofilm formation process. The adrA and sidA genes showed higher detection frequencies than csgD. Experiments analyzing the biofilm production process by S. Heidelberg strains are not common, and further studies are necessary to understand this complex process.

Phenotypic and Molecular Characterization of Salmonella Enteritidis SE86 Isolated from Poultry and Salmonellosis Outbreaks.

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.

Spread of a Major Clone of Salmonella enterica Serotype Enteritidis in Poultry and in Salmonellosis Outbreaks in Southern Brazil.

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.

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 .