Identification of Brucella sp. isolated in Brazil from 1976 to 2013 by Bruce-Ladder PCR

Background: Brucella sp. are the causative agents of brucellosis, an infectious disease that affects various species of animals and can be transmitted to humans through direct contact with infected animals, indirectly by the ingestion of rawmilk products, and during the handling of strains or infected material in the laboratory. Being a zoonosis, the detection of Brucella species in animals is essential for the prevention of the disease in humans and to perform a good program of control in infected herds. This study aimed at identifying Brucella field strains isolated from 1976 to 2013 in Brazil, using the modified Bruce-Ladder method, to evaluate the performance of this technique. Materials, Methods & Results: Eighty-three strains of Brucella sp. were included in the study, i.e. 21 reference strains (nine B. abortus, one B. canis, four B. melitensis, two B. ovis and five B. suis) and 62 field strains (six B. canis, one B. suis and 55 B. abortus). For the identification of the genus and/or species of Brucella, biochemical and physiological tests, including MacConkey-agar growth, glucose fermentation, haemolysis, catalase, oxidase and urease tests, nitrate reduction, citrate utilization, H2 S production and CO2 requirement, were performed. Genomic DNA was extracted from pure cultures through heat-lysis of bacterial cultures and the genus was confirmed by a genus-specific PCR (bcsp31 target gene), before performing the modified Bruce-Ladder PCR for the confirmation of the Brucella species. No problems of specificity were observed with the Bruce-Ladder PCR. However, the 1,682 bp fragment was not systematically amplified, even after several modifications such as the concentration of mix components, annealing temperatures and time. Therefore, an individual PCR using primers specific to this fragment was needed for complete identification of some strains. Also, only one kind of Polymerase gave the best results. All Brucella reference strains and negative controls gave the expected results. All field strains previously identified as B. abortus, B. canis and B. suis by biochemical and physiological tests were confirmed by the modified Bruce-Ladder PCR. All isolated Brucella abortus presented a Bruce-Ladder PCR profile expected for field strains, excluding the vaccine strains. Discussion: The modified Bruce-Ladder PCR identified properly all Brucella species (reference and field strains) and proved to be a reliable technique, thus facilitating the identification of the species in the laboratory, reducing the manipulation of these bacteria and the associated danger. Albeit the difficulties of amplification of one fragment for some strains, when using the multiplex technique, this method is fast and without risks after inactivation of the strains. Most studies on animal brucellosis in Brazil were only based on serological tests without identification of the pathogen; while the knowledge of the particular species and/or biovars that occur in Brazil, as well as their distribution, is important to monitor the spread of Brucella among sensitive species and among farms. Our results showed also that B. abortus is still the predominant species isolated in cattle in Brazil. The knowledge of the species that occur in Brazil can help to identify the source of infection and the measures of control to be applied, while it is also very important to trace the dispersion of strains among farms

Catálogo Taxonômico da Fauna do Brasil: setting the baseline knowledge on the animal diversity in Brazil

The limited temporal completeness and taxonomic accuracy of species lists, made available in a traditional manner in scientific publications, has always represented a problem. These lists are invariably limited to a few taxonomic groups and do not represent up-to-date knowledge of all species and classifications. In this context, the Brazilian megadiverse fauna is no exception, and the Catálogo Taxonômico da Fauna do Brasil (CTFB) (http://fauna.jbrj.gov.br/), made public in 2015, represents a database on biodiversity anchored on a list of valid and expertly recognized scientific names of animals in Brazil. The CTFB is updated in near real time by a team of more than 800 specialists. By January 1, 2024, the CTFB compiled 133,691 nominal species, with 125,138 that were considered valid. Most of the valid species were arthropods (82.3%, with more than 102,000 species) and chordates (7.69%, with over 11,000 species). These taxa were followed by a cluster composed of Mollusca (3,567 species), Platyhelminthes (2,292 species), Annelida (1,833 species), and Nematoda (1,447 species). All remaining groups had less than 1,000 species reported in Brazil, with Cnidaria (831 species), Porifera (628 species), Rotifera (606 species), and Bryozoa (520 species) representing those with more than 500 species. Analysis of the CTFB database can facilitate and direct efforts towards the discovery of new species in Brazil, but it is also fundamental in providing the best available list of valid nominal species to users, including those in science, health, conservation efforts, and any initiative involving animals. The importance of the CTFB is evidenced by the elevated number of citations in the scientific literature in diverse areas of biology, law, anthropology, education, forensic science, and veterinary science, among others

Ocorrência de Staphylococcus pseudintermedius resistente à meticilina (MRSP) em cães atendidos no hospital de clínicas veterinárias da UFRGS (HCV-UFRGS)

Staphylococcus (S.) pseudintermedius é o principal agente causador de piodermite e otite crônica canina e o isolamento desta espécie em infecções humanas sugere possível caráter zoonótico deste agente. Nos últimos anos houve crescente número de relatos de cepas de S. pseudintermedius resistentes à meticilina (MRSP), as quais frequentemente pertencem a grupos clonais disseminados no ambiente hospitalar e na comunidade. O objetivo deste estudo foi investigar a presença de MRSP em amostras clínicas provenientes de pacientes atendidos entre 2011-2012 no Hospial de Clínicas Veterinárias da Universidade Federal do Rio Grande do Sul (HCV-UFRGS). Noventa e dois isolados de Staphylococcus sp. coagulase positiva foram submetidos ao teste de triagem para identificar resistentes resistência à meticilina, através do teste de disco-difusão em ágar utilizando disco de oxacilina. Dos isolados analisados, 21,7% (20/92) apresentaram perfil de resistência fenotípica à meticilina. Os isolados resistentes à meticilina foram submetidos a PCR-RFLP para identificação da espécie e investigou-se a presença do gene mecA. A amplificação e análise do gene spa foi utilizada para identificar possíveis clones. Dentre estes, 100% (20/20) pertenciam à espécie S. pseudintermedius e tiveram a presença do gene mecA confirmada. Através da tipificação do gene spa seis diferentes repetições foram identificadas e sete tipos de spa typing, sugerindo possíveis clonalidades. Conclui-se que cepas MRSP estão presentes em pacientes atendidos no HCV-UFRGS, um fato preocupante pelo potencial zoonótico e pela dificuldade de tratamento destes pacientes. A tipificação do gene spa revelou haver um grupo clonal predominante na região.Staphylococcus pseudintermedius is the main cause of pyoderma and chronic otitis in dogs. The isolation of this specie in human infections suggests that it may be a zoonotic agent as well. Recently reports of methicillin-resistant S. pseudintermedius (MRSP) clonal groups, which have spread in hospitals and community, have been increasing. The objective of this study was to assess the occurrence of MRSP isolates in patients treated between 2011-2012 at the Veterinary Hospital of the Federal University of Rio Grande do Sul (UFRGS-HCV). Ninety-two isolates of Staphylococcus sp. coagulase positive were subjected to agar disk diffusion test to assess resistance to methicillin. From the isolates analyzed, 21.7% (20/92) presented a phenotypic resistance profile to methicillin. These isolates were then submitted to PCR-RFLP for species identification and for investigation of the mecA gene presence. Amplification and analysis of the spa gene was used to identify clones. Among these isolates, 100% (20/20) was identified as S. pseudintermedius and presented the mecA gene. In spa gene analysis six different repetitions, and seven spa types were identified. It was concluded that MRSP isolates are present in patients treated at the HCV-UFRGS, a matter of concern taking into account the difficulty of treating these patients and their zoonotic hazard. The spa gene classification showed that a MRSP clonal group is present in the region

Padronização dos critérios de avaliação adotados pelos auditores sanitários internos e educação continuada de colaboradores em uma rede de supermercados, no Estado do Rio Grande do Sul

Uma equipe de auditores em Segurança de Alimentos necessita estar padronizada em relação às suas exigências diárias. Além das auditorias, esses profissionais devem executar e avaliar a educação continuada de manipuladores de alimentos. Considerando isso, o presente estudo teve por objetivos i) a padronização dos critérios de avaliação de auditores sanitários de uma rede de supermercados e ii) a padronização da educação continuada em Procedimentos Operacionais Padronizados (POPs) de higienização com manipuladores de alimentos utilizando a leitura residual de ATP (luminômetro) como instrumento educativo. Utilizou-se amostragem do tipo não probabilística por conveniência e intencional, sendo uma parte selecionada de acordo com o acesso do pesquisador (Conveniência) e a outra parte por seleção de subgrupo (Intencional). A abordagem para a coleta dos dados junto aos auditores foi qualitativa de Grupo Focal. A primeira etapa consistiu na padronização dos critérios de avaliação com o preenchimento de um check-list de auditoria interna, baseando-se na severidade e probabilidade da não conformidade. Foram selecionadas 20 questões do check-list que apresentavam discordância de interpretação entre os auditores. Cruzando os critérios de severidade e probabilidade foram elaborados quadros descrevendo cada questão, não conformidade, probabilidade, severidade e resposta padrão esperada. Consolidados os critérios foi realizada uma simulação de check-list com os auditores, utilizando-se fotos de um banco de dados de auditorias passadas. Através do coeficiente Kappa, os gabaritos foram analisados para medir o grau de concordância entre todos. As concordâncias resultantes foram substantivas e uma categorizada como quase perfeita. Para a segunda etapa, os auditores identificaram a necessidade de criação de novas abordagens para a educação continuada em POPs de higienização com os colaboradores e acordou-se o uso do luminômetro como ferramenta educativa. Um treinamento foi aplicado em 28 unidades da rede, com cerca de 140 colaboradores entre março e junho de 2022, onde 217 leituras de ATP foram realizadas em equipamentos e utensílios. Antes das capacitações, a média de conformidades observadas em leituras de ATP era de 91,3%. Durante as capacitações, o percentual de conformidade foi de 81,6%. As hipóteses levantadas pela equipe para tal diminuição foram: i) o uso do luminômetro era visto anteriormente pelos colaboradores como um teste e, pensando desta forma, a higienização ocorria de forma diferenciada no dia da leitura de ATP; ii) alguns fornecedores de utensílios frequentemente testados foram trocados e notou-se uma variação na qualidade dos materiais; iii) a mudança no procedimento junto aos colaboradores despertou na equipe técnica novas abordagens como, por exemplo, a utilização das falhas como meio de educação. O método de educação de colaboradores acabou por se apresentar como um processo de reflexão de ensino-aprendizagem para a equipe. Assim, o estudo concluiu que: i) o teste de concordância Kappa permitiu comprovar que o padrão de avaliação está sendo seguido por todos os auditores; ii) apesar da média de conformidades ter diminuído, o processo de alinhar a educação continuada dos colaboradores foi importante para equipe refletir sobre seus métodos educativos e sobre a necessidade da mensuração de resultados para observar se o objetivo está sendo alcançado.A team of Food Safety auditors needs to be standardized in relation to their daily requirements. In addition to audits, these professionals must perform and evaluate the education of food handlers. Considering this, the present study aims to i) standardize the evaluation criteria of Food Safety auditors in a supermarket chain and ii) standardized education in hygiene Standard Operating Procedures (SOPs) of food handlers using the residual reading of ATP as an educational tool. Non-probabilistic sampling was used for convenience and intentionally, with a part selected according to the researcher's access (Convenience) and the other part by subgroup selection (Intentional). The approach for collecting data with the auditors was a qualitative Focus Group. The first step consisted of standardizing the evaluation criteria by completing an internal audit checklist, based on the severity and probability of non-compliance. Twenty questions were selected from the checklist that presented disagreement in interpretation between the auditors. Crossing the severity and probability criteria, tables were set up describing each question, non-compliance, probability, severity and the expected standard response. Once the criteria were consolidated, a checklist simulation was carried out with the auditors, using photos from a database of past audits. Using the Kappa coefficient, the responses were analyzed to measure the degree of agreement among all. The resulting agreements were substantive and one was categorized as almost perfect. For the second stage, the auditors identified the need to create new approaches for education in hygiene SOPs with employees and it was agreed to use the residual reading of ATP as an educational tool. Training was applied in 28 supermarket units, with around 140 employees between March and June 2022, where 217 ATP readings were performed on equipment and utensils. Before training, the average agreement observed in ATP readings was 91.3%. During training, the percentage of agreement was 81.6%. The hypotheses raised by the team for this reduction were: i) the use of the residual reading of ATP was previously seen by the collaborators as a test and, in this way, the hygiene occurred in a different way on the day of the ATP reading; ii) some suppliers of frequently tested utensils were changed and a variation in the quality of materials was noted; iii) the change in the procedure with the collaborators induced new approaches in the technical team, such as, for example, the use of failures as a means of education. The employee education method turned out to be a teaching-learning reflection process for the team. Thus, the study concluded that: i) the Kappa concordance test allowed proving that the evaluation standard is being followed by all auditors; ii) despite the average of agreement decrease, the process of aligning the education of employees was important for the team to reflect on its educational methods and on the need to measure results to observe whether the objective is being achieved

Piglet colibacillosis diagnosis based on multiplex polymerase chain reaction and immunohistochemistry of paraffin-embedded tissues

Enterotoxigenic Escherichia coli (ETEC) causes diarrhea in pigs, referred to as colibacillosis. The aim of this study was to optimize multiplex polymerase chain reaction (PCR) and immunohistochemistry (IHC) analyses of paraffin-embedded material to detect pathogenic E. coli strains causing colibacillosis in pigs. Multiplex PCR was optimized for fimbriae (F18, F4, F6, F5, and F41) and toxins (types A and B heat-stable toxins [STaP and STb], heat-labile toxin [LT], and type 2 Shiga toxin [STx2e]), and IHC was optimized for an anti-E. coli polyclonal antibody. Samples (132) from pigs received between 2006 and 2014 with clinical and histopathological diagnoses of colibacillosis were analyzed. E. coli was detected by IHC in 78.7%, and at least one virulence factor gene was detected in 71.2%. Pathogenic strains of ETEC with at least one fimbria and one toxin were detected in 40% of the samples in multiplex PCR. The most frequent virulence types were F18-STaP (7.5%), F18-STaP-STb (5.7%), and F4-STaP (3.8%). A statistically significant association was noted between virulence factors F4, F18, STaP, and STb and positive immunostaining results. Colibacillosis diagnosis through multiplex PCR and IHC of paraffin-embedded tissues is a practical approach, as samples can be fixed and stored for long periods before analysis

NEOTROPICAL ALIEN MAMMALS: a data set of occurrence and abundance of alien mammals in the Neotropics

Biological invasion is one of the main threats to native biodiversity. For a species to become invasive, it must be voluntarily or involuntarily introduced by humans into a nonnative habitat. Mammals were among first taxa to be introduced worldwide for game, meat, and labor, yet the number of species introduced in the Neotropics remains unknown. In this data set, we make available occurrence and abundance data on mammal species that (1) transposed a geographical barrier and (2) were voluntarily or involuntarily introduced by humans into the Neotropics. Our data set is composed of 73,738 historical and current georeferenced records on alien mammal species of which around 96% correspond to occurrence data on 77 species belonging to eight orders and 26 families. Data cover 26 continental countries in the Neotropics, ranging from Mexico and its frontier regions (southern Florida and coastal-central Florida in the southeast United States) to Argentina, Paraguay, Chile, and Uruguay, and the 13 countries of Caribbean islands. Our data set also includes neotropical species (e.g., Callithrix sp., Myocastor coypus, Nasua nasua) considered alien in particular areas of Neotropics. The most numerous species in terms of records are from Bos sp. (n = 37,782), Sus scrofa (n = 6,730), and Canis familiaris (n = 10,084); 17 species were represented by only one record (e.g., Syncerus caffer, Cervus timorensis, Cervus unicolor, Canis latrans). Primates have the highest number of species in the data set (n = 20 species), partly because of uncertainties regarding taxonomic identification of the genera Callithrix, which includes the species Callithrix aurita, Callithrix flaviceps, Callithrix geoffroyi, Callithrix jacchus, Callithrix kuhlii, Callithrix penicillata, and their hybrids. This unique data set will be a valuable source of information on invasion risk assessments, biodiversity redistribution and conservation-related research. There are no copyright restrictions. Please cite this data paper when using the data in publications. We also request that researchers and teachers inform us on how they are using the data