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IBV variants belonging to the GI-23 lineage have circulated since 1998 in the Middle East and have spread to several countries over time. In Brazil, the first report of GI-23 occurred in 2022. The study aimed to evaluate the in vivo pathogenicity of exotic variant GI-23 isolates. Biological samples were screening by real-time RT-PCR and classified in to GI-1 or G1-11 lineages. Interestingly, 47.77% were not classified in these lineages. Nine of the unclassified strains were sequenced and showed a high similarity to the GI-23 strain. All nine were isolated and three, were studied for pathogenicity. At necropsy, the main observations were the presence of mucus in the trachea and congestion in the tracheal mucosa. In addition, lesions on the tracheas showed marked ciliostasis, and the ciliary activity confirmed the high pathogenicity of isolates. This variant is highly pathogenic to the upper respiratory tract and can cause severe kidney lesions. This study confirm a circulation of GI-23 strain in the country and report, to first time, the isolation of an exotic variant of IBV in Brazil.

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Campylobacter is not routinely tested in foodborne disease investigations in Brazil. Here, we studied the occurrence of Campylobacter among other food-related bacteria commonly found in foodborne disease outbreaks reported in Rio Grande do Sul State, Southern Brazil. Seventy-two food samples were analyzed by using culture-based detection methods during the 18-month investigation of 36 foodborne disease outbreaks. The sampled foods from the foodborne disease outbreaks were all negative for Campylobacter . However, at least one of other routinely investigated foodborne-related bacteria was detected in 29.17% (21/72) of the samples. Taken together, these results suggest the need to monitor Campylobacter in foodborne diseases to detect sporadic cases caused by Campylobacter that might go unnoticed in Rio Grande do Sul.

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The COVID-19 infection, caused by SARS-CoV-2, is inequitably distributed and more lethal among populations with lower socioeconomic status. Direct contact with contaminated surfaces has been among the virus sources, as it remains infective up to days. Several disinfectants have been shown to inactivate SARS-CoV-2, but they rapidly evaporate, are flammable or toxic and may be scarce or inexistent for vulnerable populations. Therefore, we are proposing simple, easy to prepare, low-cost and efficient antiviral films, made with a widely available dishwashing detergent, which can be spread on hands and inanimate surfaces and is expected to maintain virucidal activity for longer periods than the current sanitizers. Avian coronavirus (ACoV) was used as model of the challenge to test the antivirus efficacy of the proposed films. Polystyrene petri dishes were covered with a thin layer of detergent formula. After drying, the films were exposed to different virus doses for 10 min and virus infectivity was determined using embryonated chicken eggs, and RNA virus quantification in allantoic fluids by RT-qPCR. The films inactivated the ACoV (ranging from 103.7 to 106.7 EID50), which is chemically and morphologically similar to SARS-CoV-2, and may constitute an excellent alternative to minimize the spread of COVID-19.

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Chicken is a leading source of thermotolerant Campylobacter, which triggers human foodborne enteritis. This study evaluated thermotolerant Campylobacter contamination of retail chicken in southern Brazil, using qualitative and quantitative analyses. Selective enrichment in Bolton broth for 24 and 48 h after plating onto modified charcoal-cefoperazone-deoxycholate (mCCD) agar and Preston agar was assessed. The combined results of the detection and enumeration methods revealed a frequency of 70% occurrence of thermotolerant Campylobacter in chicken samples. Campylobacter was enumerated in 60% of the samples, whereas 46% of the samples were positive in the qualitative analysis. Quantitative analysis showed average counts of 3.10 ± 0.15 log10 CFU/sample. Higher numbers of Campylobacter-positive samples were found using 24-h enrichment before plating onto Preston agar (46%) than onto mCCD agar (2%). The majority of isolated strains were identified as Campylobacter jejuni, and Campylobacter coli was also found but to a lesser extent. Subtyping revealed a clear distinction between strains isolated from different chicken sources. The enriched samples plated onto mCCD agar showed extensive spreading of nonproducing extended-spectrum ß-lactamases Proteus mirabilis that hampered the identification of Campylobacter colonies. P. mirabilis strains showed resistance to cefoperazone, trimethoprim, and polymyxin B present in broth and plate media used and were inhibited by rifampicin present in Preston agar. The results underline the effect of the spread of contaminant strains on Campylobacter cultures, which might be prevented using a recently revised International Organization for Standardization method for qualitative analysis of chicken.

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Infectious bursal disease (IBD) is an immunosuppressive viral disease of chickens, associated with severe economic losses and major threats to poultry production worldwide. Disease prevention programs rely on unequivocal identification of the pathogen, as well as vaccination programs. This study developed a sensitive, one-step, real-time, quantitative reverse transcription polymerase chain reaction (RT-qPCR) assay using a hydrolysis probe system for infectious bursal disease virus (IBDV, VP1 gene) detection and quantification, which was compared to other routinely used diagnostic methods. The assay successfully detected IBD reference viruses and field isolates. The absence of cross-reactivity was detected with negative samples or with other avian viruses in the analytical specificity test. The detection limit of this assay was 70 RNA copies. RT-qPCR was more sensitive in the detection of serially diluted IBDV isolates compared to virus isolation. For clinical samples, the sensitivity and specificity values of RT-qPCR compared to enzyme-linked immunosorbent assay (ELISA) were 97.5% and 100%, respectively, and compared to histopathology, these values were 100% and 93.94%, respectively. RT-qPCR can provide a simple and reliable assay for IBDV surveillance programs and for evaluation of control strategies.

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The vast capacity for maintenance and dissemination in the environment are major challenges for the control of Salmonella spp. in poultry farms. The aim of this study was to assess environmental contamination by non-typhoidal Salmonella in successive broiler flocks in nine commercial broiler farms integrated with three companies in the south of Brazil, for a twelve-month production period. Recycled broiler litter, feed and swabs from the evaporative cooling system pads were analyzed, and the total enterobacteria count in the litter samples was ascertained. Positive broiler houses were identified in two of the three broiler companies studied, in which non-typhoidal Salmonella were detected for the first time in the first or second flock, and recurred in the recycled litter of subsequent flocks. Feed and evaporative cooling pad swab samples were also positive in at least one of the assessed flocks. The majority of the isolates (87.5%) originating from different flocks, broiler houses and companies that were sampled were identified as S. Heidelberg, with the prevalence of one single genotype. The total enterobacteria levels in the litter diminished as the flocks progressed, but the presence of Salmonella spp. was constant over the course of time, indicating that the litter management procedures were not capable of interrupting the cycle of residual contamination. The predominance of S. Heidelberg highlights its emergence and dissemination in this region, as well as its resistance and maintenance in the environment, and reinforces the need to improve prevention and recycled litter management measures.

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A study using sentinel broiler chickens was performed to address Campylobacter persistence in litter that was reused for successive flocks. Cloacal swabs, litter, drag swabs, darkling beetles, feed, and drinking water were weekly sampled and analyzed by standard microbiological procedures. Thermotolerant Campylobacter isolated strains were confirmed by polymerase chain reaction and subtyped by pulsed-field gel electrophoresis analysis. Campylobacter was not detected in samples collected immediately after downtime between broiler flocks. However, Campylobacter-positive samples were first detected at 21 d. After Campylobacter was initially isolated from the cloacal swabs, reused litter, drag swabs, or darkling beetles, these samples remained Campylobacter positive in the following weeks until the end of the rearing period. Campylobacter-positive cloacal swabs obtained from sentinel broilers ranged from 97.3% to 100% at 42 d. All isolated strains were identified as Campylobacter jejuni. Among the subtypes identified, an indistinguishable C. jejuni strain was predominant in sentinel broilers and was also detected in the other environmental samples analyzed, suggesting a common and persistent contamination source within the flocks. Sentinel broilers may have contributed to amplify the Campylobacter level, maintaining flock and broiler house contamination until the end of the production cycle.

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Nontyphoidal Salmonella are one of the leading causes of foodborne diseases in the world. As poultry products are recognized as main sources of human salmonellosis, nontyphoidal Salmonella control has become a global issue for the poultry industry. The increasing antimicrobial resistance in poultry-related nontyphoidal Salmonella serovars is a global matter of concern. By monitoring the evolution of antimicrobial resistance, alternative treatments can be identified and possible restrictions in the treatment of systemic human salmonellosis foreseen. A meta-analysis was conducted to assess the profile and temporal evolution of the antimicrobial resistance of nontyphoidal Salmonella of poultry and human origin in Brazil, isolated in the period from 1995 to 2014. Four databases were researched; twenty-nine articles met the eligibility criteria and were included in the meta-analysis. In the nontyphoidal isolates of poultry origin, the highest levels of antimicrobial resistance were verified for sulfonamides (44.3%), nalidixic acid (42.5%), and tetracycline (35.5%). In the human-origin isolates, the resistance occurred mainly for sulfonamides (46.4%), tetracycline (36.9%), and ampicillin (23.6%). Twenty-two articles described results of antimicrobial resistance specifically for Salmonella Enteritidis, also enabling the individual meta-analysis of this serovar. For most antimicrobials, the resistance levels of Salmonella Enteritidis were lower than those found when considering all the nontyphoidal serovars. In the poultry-origin isolates, a quadratic temporal distribution was observed, with reduced resistance to streptomycin in Salmonella Enteritidis and in all nontyphoidal serovars, and a linear increase of resistance to nalidixic acid in Salmonella Enteritidis. In the human-origin isolates, a linear increase was identified in the resistance to nalidixic acid in Salmonella Enteritidis and in all the nontyphoidal isolates, and to gentamicin in Salmonella Enteritidis. Continuous monitoring of the development and spread of antimicrobial resistance could support the measurement of the consequences on poultry and human health.

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The present study analyzes the characteristics of Salmonella spp. from broiler chicken farms in Brazil. In total, 82 Salmonella spp. strains were characterized by serotyping, determining susceptibility to antimicrobials, and using pulsed-field gel electrophoresis (PFGE). Fifteen Salmonella serotypes were identified, among which Minnesota (40.24%), Infantis (14.63%), Heidelberg (7.31%), Senftenberg (6.09%), and Mbandaka (6.09%) were the most frequent. Salmonella Minnesota occurred mostly in the state of Mato Grosso do Sul and in one of the broiler companies surveyed. Approximately 60% of the strains were resistant to at least one of the antimicrobials tested. From these isolates, 17.07% were resistant to only one antimicrobial (tetracycline or streptomycin), and 9.75% were resistant to 3 or more antimicrobial classes. Thirteen resistance profiles were characterized, the most frequent of which were the resistance to tetracycline (15.85%); to the combination of trimethroprim with sulfamethoxazole, and tetracycline (10.97%); and to the combination of streptomycin and tetracycline (9.75%). Multiple correspondence analysis revealed that susceptibility or resistance of the analyzed strains and also particular Salmonella serotypes were associated with broiler-producing companies where the samples were collected. Strains presented high intraserotype genetic variability, as shown by the 64 PFGE profiles, suggesting the existence of several contamination sources in the surveyed farms.

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Mycoplasma synoviae is a Gram positive bacteria lacking of cell wall that affects chickens and turkeys causing infection in the upper respiratory tract and in some cases arthritis, with economical impact to broiler breeders. Treatment and prevention of avian synovitis depend on knowledge of the infectious process. Secreted or surface-exposed proteins play a critical role in disease because they often mediate interactions between host and pathogen. In the present work, we sought to identify possible M. synoviae secreted proteins by cultivating the bacteria in a modified protein-free Frey medium. Using this approach, we were able to detect in the cell-free fraction a number of proteins that have been shown in other organisms to be secreted, suggesting that they may also be secreted by M. synoviae.

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Background: Broilers are a reservoir of Campylobacter (C.), an important causal agent of gastroenteritis mostly associated to handling and consumption of broiler meat. The majority of broiler fl ocks are colonized by thermophilic Campylobacter at the slaughter age, and carcasses might be contaminated throughout the processing line. Since surveillance is crucial to evaluate and improve approaches to reduce Campylobacter spread during broiler processing, a cross-sectional study was carried out to detect the level of Campylobacter contamination in a broiler at slaughterMaterials, Methods & Results: Cloacal swabs, caeca and whole carcasses were taken from a broiler fl ock slaughtered in Southern Brazil. Samples were individually inoculated in Bolton Broth (BB) and incubated at 41.5C in a microaerobic atmosphere for 44 h, when the enriched culture was inoculated onto modifi ed Charcoal Cefoperazone Deoxycholate Agar (mCCDA) and Campy-Cefex Agar (CCA) plates. All plates were incubated at 41.5C in the microaerobic atmosphere for 44 h. Aliquots of each enriched BB were collected and submitted to polymerase chain reaction (PCR), while the genetic relatedness of isolates was analyzed by pulsed-fi eld gel electrophoresis (PFGE). A total of 3 (9.4%) cloacal swabs were positive for C. jejuni. No Campylobacter was isolated from any of the caecal contents or broiler carcasses analyzed.

Background: Broilers are a reservoir of Campylobacter (C.), an important causal agent of gastroenteritis mostly associated to handling and consumption of broiler meat. The majority of broiler fl ocks are colonized by thermophilic Campylobacter at the slaughter age, and carcasses might be contaminated throughout the processing line. Since surveillance is crucial to evaluate and improve approaches to reduce Campylobacter spread during broiler processing, a cross-sectional study was carried out to detect the level of Campylobacter contamination in a broiler at slaughter.Materials, Methods & Results: Cloacal swabs, caeca and whole carcasses were taken from a broiler fl ock slaughtered in Southern Brazil. Samples were individually inoculated in Bolton Broth (BB) and incubated at 41.5C in a microaerobic atmosphere for 44 h, when the enriched culture was inoculated onto modifi ed Charcoal Cefoperazone Deoxycholate Agar (mCCDA) and Campy-Cefex Agar (CCA) plates. All plates were incubated at 41.5C in the microaerobic atmosphere for 44 h. Aliquots of each enriched BB were collected and submitted to polymerase chain reaction (PCR), while the genetic relatedness of isolates was analyzed by pulsed-fi eld gel electrophoresis (PFGE). A total of 3 (9.4%) cloacal swabs were positive for C. jejuni. No Campylobacter was isolated from any of the caecal contents or broiler carcasses analyzed.

RESUMO

Background: Broilers are a reservoir of Campylobacter (C.), an important causal agent of gastroenteritis mostly associated to handling and consumption of broiler meat. The majority of broiler flocks are colonized by thermophilic Campylobacter at the slaughter age, and carcasses might be contaminated throughout the processing line. Since surveillance is crucial to evaluate and improve approaches to reduce Campylobacter spread during broiler processing, a cross-sectional study was carried out to detect the level of Campylobacter contamination in a broiler at slaughter. Materials, Methods & Results: Cloacal swabs, caeca and whole carcasses were taken from a broiler flock slaughtered in Southern Brazil. Samples were individually inoculated in Bolton Broth (BB) and incubated at 41.5°C in a microaerobic atmosphere for 44 h, when the enriched culture was inoculated onto modified Charcoal Cefoperazone Deoxycholate Agar (mCCDA) and Campy-Cefex Agar (CCA) plates. All plates were incubated at 41.5°C in the microaerobic atmosphere for 44 h. Aliquots of each enriched BB were collected and submitted to polymerase chain reaction (PCR), while the genetic relatedness of isolates was analyzed by pulsed-field gel electrophoresis (PFGE). A total of 3 (9.4%) cloacal swabs were positive for C. jejuni. No Campylobacter was isolated from any of the caecal contents or broiler carcasses analyzed. In addition, negative mCCDA and CCA plates showed an abundant growth of contaminant cells. The PCR assay detected all thermophilic Campylobacter reference strains tested and also the Arcobacter species. No amplified product was obtained from the non-related bacterial species analyzed. It was possible to identify 29 (90.6%) cloacal swabs, 32 (97.0%) caecal contents and 31 (100%) broiler carcasses Campylobacter-positive by PCR analysis. PFGE typing of the C. jejuni isolated resulted in two clearly distinguished genotypes which were grouped into different clusters. Discussion: The detection of C. jejuni in only few cloacal swabs sampled contrasts with higher frequencies of Campylobacter previously described in broilers. However, the enrichment culture of fecal samples might be compromised by the many competing non-target bacteria present, which may have prevented the detection of Campylobacter-positive samples. In addition, the BB and selective media containing cefoperazone might have allowed the growth of cefoperazone-resistant contaminant cells from fecal and carcasses samples, which masked Campylobacter cells onto mCCDA and CCA. To improve the detection of Campylobacter in broiler samples, alternative antimicrobial supplements or reduction of the time of enrichment has already been suggested. PCR showed a higher number of positive samples, which might reflect the increased ability of the PCR assay to detect either injured cells in conventional enrichment culture or Campylobacter that were masked by the proliferation of competing cells onto selective media used. The PCR assay was able to detect all the reference strains of thermophilic Campylobacter, but also the related Arcobacter species. However, the temperature of incubation of the enriched cultures associated to the selective pressure of the antimicrobials present in the BB restricts the growth of Arcobacter and the false-positive results observed using PCR. The subtypes of the C. jejuni strains isolated showed that the target broiler flock was simultaneously colonized by more than one C. jejuni strain which might be the result of introduction of Campylobacter from different sources at farm. PCR analysis showed high Campylobacter contamination level of the target flock at slaughter, pointing to the need for additional studies to investigate Campylobacter sources at broiler processing.

Assuntos

RESUMO

Background: Broilers are a reservoir of Campylobacter (C.), an important causal agent of gastroenteritis mostly associated to handling and consumption of broiler meat. The majority of broiler fl ocks are colonized by thermophilic Campylobacter at the slaughter age, and carcasses might be contaminated throughout the processing line. Since surveillance is crucial to evaluate and improve approaches to reduce Campylobacter spread during broiler processing, a cross-sectional study was carried out to detect the level of Campylobacter contamination in a broiler at slaughterMaterials, Methods & Results: Cloacal swabs, caeca and whole carcasses were taken from a broiler fl ock slaughtered in Southern Brazil. Samples were individually inoculated in Bolton Broth (BB) and incubated at 41.5C in a microaerobic atmosphere for 44 h, when the enriched culture was inoculated onto modifi ed Charcoal Cefoperazone Deoxycholate Agar (mCCDA) and Campy-Cefex Agar (CCA) plates. All plates were incubated at 41.5C in the microaerobic atmosphere for 44 h. Aliquots of each enriched BB were collected and submitted to polymerase chain reaction (PCR), while the genetic relatedness of isolates was analyzed by pulsed-fi eld gel electrophoresis (PFGE). A total of 3 (9.4%) cloacal swabs were positive for C. jejuni. No Campylobacter was isolated from any of the caecal contents or broiler carcasses analyzed.

Background: Broilers are a reservoir of Campylobacter (C.), an important causal agent of gastroenteritis mostly associated to handling and consumption of broiler meat. The majority of broiler fl ocks are colonized by thermophilic Campylobacter at the slaughter age, and carcasses might be contaminated throughout the processing line. Since surveillance is crucial to evaluate and improve approaches to reduce Campylobacter spread during broiler processing, a cross-sectional study was carried out to detect the level of Campylobacter contamination in a broiler at slaughter.Materials, Methods & Results: Cloacal swabs, caeca and whole carcasses were taken from a broiler fl ock slaughtered in Southern Brazil. Samples were individually inoculated in Bolton Broth (BB) and incubated at 41.5C in a microaerobic atmosphere for 44 h, when the enriched culture was inoculated onto modifi ed Charcoal Cefoperazone Deoxycholate Agar (mCCDA) and Campy-Cefex Agar (CCA) plates. All plates were incubated at 41.5C in the microaerobic atmosphere for 44 h. Aliquots of each enriched BB were collected and submitted to polymerase chain reaction (PCR), while the genetic relatedness of isolates was analyzed by pulsed-fi eld gel electrophoresis (PFGE). A total of 3 (9.4%) cloacal swabs were positive for C. jejuni. No Campylobacter was isolated from any of the caecal contents or broiler carcasses analyzed.

RESUMO

in vitro antibacterial activity of 21 hydroethanolic vegetal extracts was assessed against 20 serovars of Salmonella. Regarding the tested extracts, 85.7 percent of them presented antibacterial activity. The six active extracts which showed activity on the largest number of serovars and the extract of Eucalyptus sp. were submitted to the determination of Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC). Of these, six extracts showed bacteriostatic and bactericidal activity with MIC and MBC for Punica granatum (pomegranate) from 20 and 60mg mL-1, for Eugenia jambolana (rose apple) from 40 and 240mg mL-1, Eugenia uniflora (surinam cherry) from 80 and 240mg mL-1, Caryophyllus aromaticus (clove) from 10 and 60mg mL-1, Psidium araca from 30 and 320mg mL-1 and Eucalyptus sp. from 40 and 160mg mL-1. Achyrocline satureioides (macela) presented only bacteriostatic potential and MIC from 160mg mL-1. Caryophyllus aromaticus, Eucalyptus sp., and Psidium araca presented the best results for bactericidal activity, inhibiting, respectively, 84.2 percent, 42.1 percent, and 17.6 percent of Salmonella's serovars. The activity of each extract varied for different serovars; S. London presented resistance to the six extracts in MBC, while S. Pullorum was the most susceptible serovar.

A atividade antibacteriana de 21 extratos hidroetanólicos vegetais foi avaliada in vitro frente a 20 sorovares de Salmonella. Dos extratos testados, 85,7 por cento apresentaram atividade antibacteriana. Os seis extratos que evidenciaram atividade sobre o maior número de sorovares e Eucalyptus sp. foram submetidos à determinação da Concentração Inibitória Mínima (CIM) e Concentração Bactericida Mínima (CBM). Destes, seis extratos apresentaram atividade bacteriostática e bactericida com MIC para Punica granatum (romã) a partir de 20 e 60mg mL-1, Eugenia jambolana (jambolão) de 40 e 240mg mL-1, Eugenia uniflora (pitanga) de 80 e 240mg mL-1, Caryophyllus aromaticus (cravo) de 10 e 60mg mL-1, Psidium araca (araçá) 30 e 320mg mL-1 e Eucalyptus sp. (eucalipto) de 40 e 160mg mL-1. Achyrocline satureioides (macela) apresentou apenas atividade bacteriostática e MIC a partir de 160mg mL-1. Caryophyllus aromaticus, Eucalyptus sp. e Psidium araca apresentaram os melhores resultados para a atividade bactericida, inativando, respectivamente, 84,21 por cento, 42,1 por cento e 17,64 por cento dos sorovares de Salmonella. A atividade de cada extrato variou para diferentes sorovares. Nenhum dos seis extratos avaliados evidenciou atividade bactericida frente a S. London, enquanto S. Pullorum foi o sorovar mais sensível.

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Background: Human campylobacteriosis has become a major foodborne disease, although little information is available about the role it plays in the scenario of foodborne diseases in Brazil. Since thermophilic Campylobacter (C.) species are often found in the intestinal tract of broiler chickens, consumption of contaminated poultry meat has been considered a risk factor for Campylobacter human infection. Campylobacter has been described as extremely susceptible to a variety of environmental stresses; hence the difficulty to establish cultures of the microorganism in the laboratory. In addition, it has been shown to decline in refrigerated and frozen foods. Currently there is a need for data on Campylobacter contamination level in Brazilian poultry meat. This work describes a survey performed in chilled and frozen poultry meat obtained from different retailers in Concórdia, Santa Catarina, Brazil. Materials, Methods & Results: This study analyzed 24 samples of fresh (chilled or frozen) poultry meat portions (thighs and drumsticks) produced by three different Brazilian broiler chicken processors, which were purchased from three different retailers in Concórdia, Santa Catarina. Campylobacter isolation was performed according to ISO 10272-1:2006. Individual samples (25g) were enriched in 225 mL of Bolton Broth in microaerobic atmosphere at 37°C for 4h to 6h, then at 41.5°C for 44h (+/- 4h). Aliquots were streaked in Modif ed Charcoal Cefoperazone Deoxycholate Agar (mCCDA) and Campy-Cefex Agar at 41.5°C in a microaerobic atmosphere for 44h (+/- 4h). Suspected colonies were subcultured in Blood Agar no 2 plates for confirmation by morphology, Gram staining, tests for catalase, oxidase and hippurate and indoxyl acetate hydrolysis test. Enriched Bolton Broth aliquots were also analyzed by Polymerase Chain Reaction (PCR) to amplify a 287 bp sequence of the 16S rRNA gene from thermophilic Campylobacter. Although C. jejuni positive control was isolated and confirmed by the morphological and biochemical tests described, all samples analyzed were negative for the presence of thermophilic Campylobacter. However, either mCCDA or Campy-Cefex Agar showed an abundant growth of non-Campylobacter cells. Moreover, all samples analyzed were negative in PCR analysis, although the C. jejuni reference strain used as PCR positive control showed the expected DNA fragment amplified. Discussion: Higher levels of Campylobacter contamination in poultry meat have been found by other studies, which used different isolation protocols in comparison to the present work. This study shows that Campylobacter isolation procedure according to ISO 10272-1:2006 allowed the growth of contaminants in both selective media used. It might be result of enrichment for 48 h or proliferation of ESBL producing Escherichia coli, able to hydrolyze cefoperazone in the selective medium used, which could underestimate the presence of Campylobacter in samples. In this sense, the PCR assay was essential to corroborate the negative result in Campylobacter isolation. Because poultry meat analyzed was negative for the presence of thermophilic Campylobacter, it was not possible to assess any difference between chilled or frozen storage on the bacteria survival. The present study might reflect low rates of Campylobacter contamination in poultry meat available at retail in Concórdia, Santa Catarina, Brazil, although it cannot replace good hygienic practices as well as consumer education.

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Background: Human campylobacteriosis has become a major foodborne disease, although little information is available about the role it plays in the scenario of foodborne diseases in Brazil. Since thermophilic Campylobacter (C.) species are often found in the intestinal tract of broiler chickens, consumption of contaminated poultry meat has been considered a risk factor for Campylobacter human infection. Campylobacter has been described as extremely susceptible to a variety of environmental stresses; hence the difficulty to establish cultures of the microorganism in the laboratory. In addition, it has been shown to decline in refrigerated and frozen foods. Currently there is a need for data on Campylobacter contamination level in Brazilian poultry meat. This work describes a survey performed in chilled and frozen poultry meat obtained from different retailers in Concórdia, Santa Catarina, Brazil. Materials, Methods & Results: This study analyzed 24 samples of fresh (chilled or frozen) poultry meat portions (thighs and drumsticks) produced by three different Brazilian broiler chicken processors, which were purchased from three different retailers in Concórdia, Santa Catarina. Campylobacter isolation was performed according to ISO 10272-1:2006. Individual samples (25g) were enriched in 225 mL of Bolton Broth in microaerobic atmosphere at 37°C for 4h to 6h, then at 41.5°C for 44h (+/- 4h). Aliquots were streaked in Modif ed Charcoal Cefoperazone Deoxycholate Agar (mCCDA) and Campy-Cefex Agar at 41.5°C in a microaerobic atmosphere for 44h (+/- 4h). Suspected colonies were subcultured in Blood Agar no 2 plates for confirmation by morphology, Gram staining, tests for catalase, oxidase and hippurate and indoxyl acetate hydrolysis test. Enriched Bolton Broth aliquots were also analyzed by Polymerase Chain Reaction (PCR) to amplify a 287 bp sequence of the 16S rRNA gene from thermophilic Campylobacter. Although C. jejuni positive control was isolated and confirmed by the morphological and biochemical tests described, all samples analyzed were negative for the presence of thermophilic Campylobacter. However, either mCCDA or Campy-Cefex Agar showed an abundant growth of non-Campylobacter cells. Moreover, all samples analyzed were negative in PCR analysis, although the C. jejuni reference strain used as PCR positive control showed the expected DNA fragment amplified. Discussion: Higher levels of Campylobacter contamination in poultry meat have been found by other studies, which used different isolation protocols in comparison to the present work. This study shows that Campylobacter isolation procedure according to ISO 10272-1:2006 allowed the growth of contaminants in both selective media used. It might be result of enrichment for 48 h or proliferation of ESBL producing Escherichia coli, able to hydrolyze cefoperazone in the selective medium used, which could underestimate the presence of Campylobacter in samples. In this sense, the PCR assay was essential to corroborate the negative result in Campylobacter isolation. Because poultry meat analyzed was negative for the presence of thermophilic Campylobacter, it was not possible to assess any difference between chilled or frozen storage on the bacteria survival. The present study might reflect low rates of Campylobacter contamination in poultry meat available at retail in Concórdia, Santa Catarina, Brazil, although it cannot replace good hygienic practices as well as consumer education.(AU)

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in vitro antibacterial activity of 21 hydroethanolic vegetal extracts was assessed against 20 serovars of Salmonella. Regarding the tested extracts, 85.7% of them presented antibacterial activity. The six active extracts which showed activity on the largest number of serovars and the extract of Eucalyptus sp. were submitted to the determination of Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC). Of these, six extracts showed bacteriostatic and bactericidal activity with MIC and MBC for Punica granatum (pomegranate) from 20 and 60mg mL-1, for Eugenia jambolana (rose apple) from 40 and 240mg mL-1, Eugenia uniflora (surinam cherry) from 80 and 240mg mL-1, Caryophyllus aromaticus (clove) from 10 and 60mg mL-1, Psidium araca from 30 and 320mg mL-1 and Eucalyptus sp. from 40 and 160mg mL-1. Achyrocline satureioides (macela) presented only bacteriostatic potential and MIC from 160mg mL-1. Caryophyllus aromaticus, Eucalyptus sp., and Psidium araca presented the best results for bactericidal activity, inhibiting, respectively, 84.2%, 42.1%, and 17.6% of Salmonella's serovars. The activity of each extract varied for different serovars; S. London presented resistance to the six extracts in MBC, while S. Pullorum was the most susceptible serovar.

A atividade antibacteriana de 21 extratos hidroetanólicos vegetais foi avaliada in vitro frente a 20 sorovares de Salmonella. Dos extratos testados, 85,7% apresentaram atividade antibacteriana. Os seis extratos que evidenciaram atividade sobre o maior número de sorovares e Eucalyptus sp. foram submetidos à determinação da Concentração Inibitória Mínima (CIM) e Concentração Bactericida Mínima (CBM). Destes, seis extratos apresentaram atividade bacteriostática e bactericida com MIC para Punica granatum (romã) a partir de 20 e 60mg mL-1, Eugenia jambolana (jambolão) de 40 e 240mg mL-1, Eugenia uniflora (pitanga) de 80 e 240mg mL-1, Caryophyllus aromaticus (cravo) de 10 e 60mg mL-1, Psidium araca (araçá) 30 e 320mg mL-1 e Eucalyptus sp. (eucalipto) de 40 e 160mg mL-1. Achyrocline satureioides (macela) apresentou apenas atividade bacteriostática e MIC a partir de 160mg mL-1. Caryophyllus aromaticus, Eucalyptus sp. e Psidium araca apresentaram os melhores resultados para a atividade bactericida, inativando, respectivamente, 84,21%, 42,1% e 17,64% dos sorovares de Salmonella. A atividade de cada extrato variou para diferentes sorovares. Nenhum dos seis extratos avaliados evidenciou atividade bactericida frente a S. London, enquanto S. Pullorum foi o sorovar mais sensível.

RESUMO

in vitro antibacterial activity of 21 hydroethanolic vegetal extracts was assessed against 20 serovars of Salmonella. Regarding the tested extracts, 85.7% of them presented antibacterial activity. The six active extracts which showed activity on the largest number of serovars and the extract of Eucalyptus sp. were submitted to the determination of Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC). Of these, six extracts showed bacteriostatic and bactericidal activity with MIC and MBC for Punica granatum (pomegranate) from 20 and 60mg mL-1, for Eugenia jambolana (rose apple) from 40 and 240mg mL-1, Eugenia uniflora (surinam cherry) from 80 and 240mg mL-1, Caryophyllus aromaticus (clove) from 10 and 60mg mL-1, Psidium araca from 30 and 320mg mL-1 and Eucalyptus sp. from 40 and 160mg mL-1. Achyrocline satureioides (macela) presented only bacteriostatic potential and MIC from 160mg mL-1. Caryophyllus aromaticus, Eucalyptus sp., and Psidium araca presented the best results for bactericidal activity, inhibiting, respectively, 84.2%, 42.1%, and 17.6% of Salmonella's serovars. The activity of each extract varied for different serovars; S. London presented resistance to the six extracts in MBC, while S. Pullorum was the most susceptible serovar.

A atividade antibacteriana de 21 extratos hidroetanólicos vegetais foi avaliada in vitro frente a 20 sorovares de Salmonella. Dos extratos testados, 85,7% apresentaram atividade antibacteriana. Os seis extratos que evidenciaram atividade sobre o maior número de sorovares e Eucalyptus sp. foram submetidos à determinação da Concentração Inibitória Mínima (CIM) e Concentração Bactericida Mínima (CBM). Destes, seis extratos apresentaram atividade bacteriostática e bactericida com MIC para Punica granatum (romã) a partir de 20 e 60mg mL-1, Eugenia jambolana (jambolão) de 40 e 240mg mL-1, Eugenia uniflora (pitanga) de 80 e 240mg mL-1, Caryophyllus aromaticus (cravo) de 10 e 60mg mL-1, Psidium araca (araçá) 30 e 320mg mL-1 e Eucalyptus sp. (eucalipto) de 40 e 160mg mL-1. Achyrocline satureioides (macela) apresentou apenas atividade bacteriostática e MIC a partir de 160mg mL-1. Caryophyllus aromaticus, Eucalyptus sp. e Psidium araca apresentaram os melhores resultados para a atividade bactericida, inativando, respectivamente, 84,21%, 42,1% e 17,64% dos sorovares de Salmonella. A atividade de cada extrato variou para diferentes sorovares. Nenhum dos seis extratos avaliados evidenciou atividade bactericida frente a S. London, enquanto S. Pullorum foi o sorovar mais sensível.