Publications by authors named "Pedro Panzenhagen"

13 Publications

  • Page 1 of 1

Antimicrobial Resistance Gene Detection Methods for Bacteria in Animal-Based Foods: A Brief Review of Highlights and Advantages.

Microorganisms 2021 Apr 26;9(5). Epub 2021 Apr 26.

Molecular & Analytical Laboratory Center, Faculty of Veterinary, Department of Food Technology, Universidade Federal Fluminense, Niterói 24220-900, Brazil.

Antimicrobial resistance is a major public health problem and is mainly due to the indiscriminate use of antimicrobials in human and veterinary medicine. The consumption of animal-based foods can contribute to the transfer of these genes between animal and human bacteria. Resistant and multi-resistant bacteria such as spp. and spp. have been detected both in animal-based foods and in production environments such as farms, industries and slaughterhouses. This review aims to compile the techniques for detecting antimicrobial resistance using traditional and molecular methods, highlighting their advantages and disadvantages as well as the effectiveness and confidence of their results.
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http://dx.doi.org/10.3390/microorganisms9050923DOI Listing
April 2021

Virulence genes identification and characterization revealed the presence of the Yersinia High Pathogenicity Island (HPI) in Salmonella from Brazil.

Gene 2021 Jun 20;787:145646. Epub 2021 Apr 20.

Molecular & Analytical Laboratory Center, Faculty of Veterinary, Department of Food Technology, Universidade Federal Fluminense, Niterói, Brazil; Chemistry Institute, Food Science Program, Universidade Federal do Rio de Janeiro, Rio de Janeiro, Brazil; Center for Food Analysis (NAL-LADETEC), Universidade Federal do Rio de Janeiro, Rio de Janeiro, Brazil; National Institute of Health Quality Control, Fundação Oswaldo Cruz, Rio de Janeiro, Brazil.

Salmonella spp. is one of the major agents of foodborne disease worldwide, and its virulence genes are responsible for the main pathogenic mechanisms of this micro-organism. The whole-genome sequencing (WGS) of pathogens has become a lower-cost and more accessible genotyping tool providing many gene analysis possibilities. This study provided an in silico investigation of 129 virulence genes, including plasmidial and bacteriophage genes from Brazilian strains' public Salmonella genomes. The frequency analysis of the four most sequenced serovars and a temporal analysis over the past four decades was also performed. The NCBI sequence reads archive (SRA) database comprised 1077 Salmonella public whole-genome sequences of strains isolated in Brazil between 1968 and 2018. Among the 1077 genomes, 775 passed in Salmonella in silico Typing (SISTR) quality control, which also identified 41 different serovars in which the four most prevalent were S. Enteritidis, S. Typhimurium, S. Dublin, and S. Heidelberg. Among these, S. Heidelberg presented the most distinct virulence profile, besides presenting Yersinia High Pathogenicity Island (HPI), rare and first reported in Salmonella from Brazil. The genes mgtC, csgC, ssaI and ssaS were the most prevalent within the 775 genomes with more than 99% prevalence. On the other hand, the less frequent genes were astA, iucBCD, tptC and shdA, with less than 1% frequency. All of the plasmids and bacteriophages virulence genes presented a decreasing trend between the 2000 s and 2010 s decades, except for the phage gene grvA, which increased in this period. This study provides insights into Salmonella virulence genes distribution in Brazil using freely available bioinformatics tools. This approach could guide in vivo and in vitro studies besides being an interesting method for the investigation and surveillance of Salmonella virulence. Moreover, here we propose the genes mgtC, csgC, ssaI and ssaS as additional targets for PCR identification of Salmonella in Brazil due to their very high frequency in the studied genomes.
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http://dx.doi.org/10.1016/j.gene.2021.145646DOI Listing
June 2021

Genetic diversity and multidrug-resistance among Salmonella Typhimurium isolated from swine carcasses and slaughterhouses in Rio de Janeiro, Brazil.

Vet Ital 2020 Dec 1;56(4):245-250. Epub 2020 Dec 1.

Universidade Federal Fluminense.

This study, conducted in the State of Rio de Janeiro, aimed to genetically distinguish 29 isolates of S. Typhimurium isolated from 344 samples of swine carcasses by PFGE (pulse‑field gel electrophoresis) and to evaluate their antimicrobial resistance profile. Out of the 29 isolates, 26 (90%) were resistant to at least one antimicrobial. Sulfonamides (66%), nalidixic acid (66%), trimethoprim (66%) and tetracycline (52%) were the most frequent resistant drugs. Multidrug‑resistance (MDR) profile was frequent (60% of isolates). The profile Eno‑Na‑Nxn‑Fc‑C‑S‑Gm‑G‑T‑Te (14%), Cp‑Eno‑Na‑Fc‑C‑S‑G‑T‑Te (10%) and Na‑G‑T (7%) were the most frequent. Five isolates within the predominant PFGE pulsetype came from lymph nodes of distinct animals from multiple slaughterhouses indicating that this particular clone might be widespread in the Rio de Janeiro State. This paper reveals a threat to the population in the entire State and highlights the necessity of the strict control in the use of antimicrobials in swine production in the entire country.
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http://dx.doi.org/10.12834/VetIt.1688.8954.1DOI Listing
December 2020

Frequency of Antimicrobial Resistance Genes in From Brazil by Whole-Genome Sequencing Analysis: An Overview of the Last Four Decades.

Front Microbiol 2020 7;11:1864. Epub 2020 Aug 7.

Nucleus of Food Analysis (NAL), Laboratory for the Support of Technological Development (LADETEC), Chemistry Institute, Department of Chemistry, Federal University of Rio de Janeiro, Rio de Janeiro, Brazil.

is a leading human pathogen and a significant public health concern worldwide. Massive food production and distribution have contributed to this pathogen dissemination, which, combined with antimicrobial resistance (AMR), creates new control challenges in food safety. The development of AMR is a natural phenomenon and can occur in the bacterial evolutionary process. However, the overuse and the misuse of antimicrobial drugs in humans and in animals have increased AMR selective pressure. In Brazil, there is an accuracy lack in AMR frequency in because too many isolates are under-investigated for genetic and phenotypic AMR by the Brazilian health authorities and the research community. This underreporting situation makes the comprehension of the real level of AMR in the country difficult. The present study aimed to use bioinformatics tools for a rapid screening of the genetic antimicrobial resistance profile of through whole-genome sequences (WGS). A total of 930 whole-genome sequences of were retrieved from the public database of the National Biotechnology Information Center (NCBI). A total of 65 distinct resistance genes were detected, and the most frequent ones were (A), , and . Nine point mutations were detected in total, and at the 57 position (threonine → serine) was the highest frequent substitution (26.7%, 249/930), followed by at the 83 position (serine → phenylalanine) (20.0%, 186/930) and at the 87 position (aspartic acid → asparagine) (15.7%, 146/930). The prediction of resistance phenotype showed that 58.0% (540/930) of the strains can display a multidrug resistance (MDR) profile. Ciprofloxacin and nalidixic acid were the antimicrobial drugs with the highest frequency rates of the predicted phenotype resistance among the strains. The temporal analysis through the last four decades showed increased frequency rates of antimicrobial resistance genes and predicted resistance phenotypes in the 2000s and the 2010s when compared with the 1980s and 1990s. The results presented herein contributed significantly to the understanding of the strategic use of WGS associated with analysis and the predictions for the determination of AMR in from Brazil.
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http://dx.doi.org/10.3389/fmicb.2020.01864DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7426471PMC
August 2020

Antimicrobial resistance genes in bacteria from animal-based foods.

Adv Appl Microbiol 2020 23;112:143-183. Epub 2020 May 23.

Molecular and Analytical Laboratory Center, Department of Food Technology, Faculty of Veterinary, Universidade Federal Fluminense, Niterói, Brazil; Chemistry Institute, Food Science Program, Universidade Federal do Rio de Janeiro, Rio de Janeiro, Brazil; National Institute of Health Quality Control, Fundação Oswaldo Cruz, Rio de Janeiro, Brazil.

Antimicrobial resistance is a worldwide public health threat. Farm animals are important sources of bacteria containing antimicrobial resistance genes (ARGs). Although the use of antimicrobials in aquaculture and livestock has been reduced in several countries, these compounds are still routinely applied in animal production, and contribute to ARGs emergence and spread among bacteria. ARGs are transmitted to humans mainly through the consumption of products of animal origin (PAO). Bacteria can present intrinsic resistance, and once antimicrobials are administered, this resistance may be selected and multiply. The exchange of genetic material is another mechanism used by bacteria to acquire resistance. Some of the main ARGs found in bacteria present in PAO are the bla, mcr-1, cfr and tet genes, which are directly associated to antibiotic resistance in the human clinic.
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http://dx.doi.org/10.1016/bs.aambs.2020.03.001DOI Listing
April 2021

Antimicrobial Resistance in Nontyphoidal Isolates from Human and Swine Sources in Brazil: A Systematic Review of the Past Three Decades.

Microb Drug Resist 2020 Oct 15;26(10):1260-1270. Epub 2020 May 15.

Institute of Chemistry, Federal University of Rio de Janeiro, Cidade Universitária, Rio de Janeiro, Brazil.

is the leading cause of foodborne illnesses worldwide. The widespread use of antimicrobials as prophylactic, therapeutic, and growth promoters in both livestock and human medicine has resulted in selective pressure regarding antimicrobial-resistant (AMR) bacteria. This systematic review summarizes phenotypic antimicrobial resistance profiles in isolates from human and swine sources between 1990 and 2018 in Brazil. The 20 studies that matched the eligibility criteria-isolates from pigs and humans from Brazil, between 1990 and 2016, containing information on the number of isolates, and applying the disk diffusion susceptibility method-were included. During the assessed period, strains isolated from swine sources displayed the highest resistance rates for tetracycline (20.3%) and sulfonamides (17.4%). In contrast, human isolates displayed the highest resistance rates against ampicillin (19.8%) and tetracycline (17%). Typhimurium was the most frequent AMR isolate from both swine and human sources, corresponding to 67% of all isolates. From 2001 to 2005, tetracycline and ampicillin were the top antimicrobial resistance compounds, and the most frequently detected in swine and human sources, respectively. A total of 63 and 58 multiple drug resistance profiles were identified in swine and human isolates, respectively. Antimicrobial resistance has decreased throughout the 1990-2016 period, except for gentamicin and nalidixic acid in swine and human isolates, respectively. The results indicate that isolated from human and swine display resistance against clinically important antimicrobials, indicating that swine are possibly one of the main vectors for spreading human salmonellosis in Brazil.
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http://dx.doi.org/10.1089/mdr.2019.0475DOI Listing
October 2020

Draft Genome Sequences of Leptospira interrogans Serovar Copenhageni Strains Isolated from Patients with Weil's Disease in Brazil.

Microbiol Resour Announc 2020 Feb 13;9(7). Epub 2020 Feb 13.

Bacterial Zoonosis Laboratory, Oswaldo Cruz Institute, Oswaldo Cruz Foundation, Rio de Janeiro, Brazil

Leptospirosis is a worldwide zoonosis caused by pathogenic species of In Brazil, this disease is endemic, presenting epidemic potential in rainy seasons. Here, we announce the whole-genome sequences of two serovar Copenhageni strains isolated from blood samples from two icteric patients associated with severe leptospirosis in Brazil.
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http://dx.doi.org/10.1128/MRA.01538-19DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7019067PMC
February 2020

Draft Genome Sequences of 11 Serovar Typhimurium Strains Isolated from Human Systemic and Nonsystemic Sites in Brazil.

Genome Announc 2018 Feb 1;6(5). Epub 2018 Feb 1.

Department of Veterinary Microbiology and Pathology, Washington State University, Pullman, Washington, USA

serovar Typhimurium strains isolated from systemic sites outside sub-Saharan Africa have been rarely sequenced. Here, we report the draft genome sequences of  Typhimurium sequence type 19 (ST19) ( = 9), ST1649 ( = 1), and ST313 ( = 1) strains isolated from human systemic (e.g., blood) and nonsystemic (e.g., stool and wounds) sites in Brazil.
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http://dx.doi.org/10.1128/genomeA.01223-17DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5794934PMC
February 2018

Genetically distinct lineages of Salmonella Typhimurium ST313 and ST19 are present in Brazil.

Int J Med Microbiol 2018 Mar 31;308(2):306-316. Epub 2018 Jan 31.

Department of Veterinary Microbiology and Pathology, Washington State University, Pullman, WA 99164, USA. Electronic address:

In sub-Saharan Africa, two genetically distinct lineages of multi-drug resistant non-typhoidal Salmonella (NTS) serovar Typhimurium sequence type 313 (ST313) are known to cause invasive disease among people. S. Typhimurium ST313 has evolved to become more human-adapted and is commonly isolated from systemic sites (eg., blood) from febrile patients in sub-Saharan Africa. Epidemiological studies indicate that S. Typhimurium is frequently isolated from systemic sites from human patients in Brazil, however, it is currently unknown if this pathogen has also evolved to become more invasive and human-adapted in this country. Here we determined genotypic and phenotypic divergence among clinical S. Typhimurium strains isolated from systemic and non-systemic sites from human patients in Brazil. We report that a subset (8/38, 20%) of epidemiologically diverse human clinical strains of S. Typhimurium recovered from systemic sites in Brazil show significantly higher intra-macrophage survival, indicating that this subset is likely more invasive. Using the whole genome sequencing and phylogenetic approaches, we identified S. Typhimurium ST313-lineage in Brazil that is genetically and phenotypically distinct from the known African ST313-lineages. We also report the identification of S. Typhimurium ST19-lineage in Brazil that is evolving similar to ST313 lineages from Africa but is genetically and phenotypically distinct from ST19-lineage commonly associated with the gastrointestinal disease worldwide. The identification of new S. Typhimurium ST313 and ST19 lineages responsible for human illnesses in Brazil warrants further epidemiological investigations to determine the incidence and spread of a genetically divergent population of this important human pathogen.
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http://dx.doi.org/10.1016/j.ijmm.2018.01.005DOI Listing
March 2018

Phenotypic and Genotypic Eligible Methods for Typhimurium Source Tracking.

Front Microbiol 2017 22;8:2587. Epub 2017 Dec 22.

Molecular and Analytical Laboratory Center, Department of Food Technology, Faculty of Veterinary, Universidade Federal Fluminense, Niterói, Brazil.

Salmonellosis is one of the most common causes of foodborne infection and a leading cause of human gastroenteritis. Throughout the last decade, serotype Typhimurium (ST) has shown an increase report with the simultaneous emergence of multidrug-resistant isolates, as phage type DT104. Therefore, to successfully control this microorganism, it is important to attribute salmonellosis to the exact source. Studies of source attribution have been performed to determine the main food/food-production animals involved, toward which, control efforts should be correctly directed. Hence, the election of a ST subtyping method depends on the particular problem that efforts must be directed, the resources and the data available. Generally, before choosing a molecular subtyping, phenotyping approaches such as serotyping, phage typing, and antimicrobial resistance profiling are implemented as a screening of an investigation, and the results are computed using frequency-matching models (i.e., Dutch, Hald and Asymmetric Island models). Actually, due to the advancement of molecular tools as PFGE, MLVA, MLST, CRISPR, and WGS more precise results have been obtained, but even with these technologies, there are still gaps to be elucidated. To address this issue, an important question needs to be answered: what are the currently suitable subtyping methods to source attribute ST. This review presents the most frequently applied subtyping methods used to characterize ST, analyses the major available microbial subtyping attribution models and ponders the use of conventional phenotyping methods, as well as, the most applied genotypic tools in the context of their potential applicability to investigates ST source tracking.
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http://dx.doi.org/10.3389/fmicb.2017.02587DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5744012PMC
December 2017

Contamination of Carcasses and Utensils in Small Swine Slaughterhouses by Salmonella in the Northwestern Region of the State of Rio de Janeiro, Brazil.

J Food Prot 2017 07;80(7):1128-1132

1 Departamento de Tecnologia de Alimentos, Faculdade de Veterinária, Universidade Federal Fluminense, Rua Vital Brasil Filho, 62, 24230-340, Niterói, Rio de Janeiro, Brazil.

Salmonella is a major foodborne pathogen that constantly threatens food safety in developed countries and underdeveloped countries such as Brazil, where it is responsible for 38% of notified cases of foodborne illness. Swine are one of the main meat-producing species that may asymptomatically carry Salmonella, periodically shedding the bacteria through feces. The state of Rio de Janeiro is not a major producer of swine meat, but small slaughterhouses are operational and produce meat for consumption within the state, although few studies have been conducted in the region. In this context, this study was designed to evaluate Salmonella contamination in carcasses, lymph nodes, feces, utensils, and the environment of three small slaughterhouses in the state of Rio de Janeiro. A total of 344 samples from carcasses, utensils, equipment, water, and the environment were collected from these slaughterhouses in the northwestern region of Rio de Janeiro. Salmonella was isolated from 36 (10.5%) samples: 10 of 48 carcasses, 19 of 96 lymph nodes, 4 of 48 fecal samples, 2 of 6 water samples, and 1 of 6 bleeding knives; 55 isolates were recovered. Serotyping revealed the predominance of Salmonella Typhimurium (20 isolates) followed by Salmonella Abony (10 isolates), Salmonella Give (7 isolates), Salmonella Heidelberg (4 isolates), and Salmonella Infantis (1 isolate). Ten isolates were only partially typeable, with only their O antigen identified, and three isolates had rough, nontypeable colonies. Despite the overall low prevalence of Salmonella in this study, all three slaughterhouses had poor hygienic and sanitary conditions, providing easy routes for carcass and, consequently, meat contamination. Thus, it is imperative to enforce sanitary inspections in these establishments and to apply good manufacture practices to assure the safety of the produced pork.
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http://dx.doi.org/10.4315/0362-028X.JFP-16-387DOI Listing
July 2017

Draft Genome Sequences of subsp. Serovar Typhimurium Strains Isolated from Chicken and Swine Carcasses in Two Distinct Geographical Regions from Rio de Janeiro State, Brazil.

Genome Announc 2017 Apr 27;5(17). Epub 2017 Apr 27.

Food Science Program, Chemistry Institute, University Federal of Rio de Janeiro (UFRJ), Rio de Janeiro, Brazil.

subsp. serovar Typhimurium is a surveyed worldwide serotype with well-characterized genomes for several different strains. In Brazil, very few studies have submitted whole-genome sequences to GenBank. This genome may be useful to analyze the genetic mechanisms comparable to those of other related studies conducted in Brazil and globally.
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http://dx.doi.org/10.1128/genomeA.00197-17DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5408100PMC
April 2017

Investigation of enrofloxacin residues in broiler tissues using ELISA and LC-MS/MS.

Food Addit Contam Part A Chem Anal Control Expo Risk Assess 2016 1;33(4):639-43. Epub 2016 Mar 1.

a Department of Veterinary Public Health and Public Health , Federal Fluminense University (UFF) , Niterói , Brazil.

This study investigated the efficiency of an enrofloxacin ELISA test kit to detect the presence of enrofloxacin residues in broiler tissues compared with LC-MS/MS. Broiler tissues from 72 samples consisting of 60 breast muscle, six pools of livers (500 g each) and six pools of kidneys (500 g each) were obtained from six different slaughterhouses. Breast muscle from 10 carcasses and pools of livers and kidneys from approximately 200 carcasses of the same flock were collected from each slaughterhouse. ELISA and HPLC were used to identify and quantify the contamination of the samples with enrofloxacin. A total of 72% of the analysed samples contained enrofloxacin residues detected by the ELISA and 22.2% were detected by LC-MS/MS. The mean values of enrofloxacin contamination found in chicken breast by ELISA and HPLC were 8.63 and 12.25 μg kg(-1), respectively. None of the samples exceeded the maximum limit of 100 μg kg(-1) by both methods set by the European Union as well as the Brazilian Agriculture Ministry. All positive samples for enrofloxacin residues detected by LC-MS/MS were also positive by ELISA. These data confirm the efficiency of the ELISA test, and suggest its use as a screening method for enrofloxacin residues in poultry tissues due to its quick results, low price and ease of applicability.
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http://dx.doi.org/10.1080/19440049.2016.1143566DOI Listing
December 2016