A realistic model for vasectomy reversal training using swine testicles

Purpose: To evaluate the viability of the porcine vas deferens as a realistic microsurgical training model for vasectomy reversal Methods: The model uses swine testicles (vas deferent), which are usually discarded in large street markets since they are not part of Brazilian cuisine. The spermatic cord was carefully dissected, and the vas deferens were isolated, measuring 10 cm in length. A paper quadrilateral with 5 cm2 was built to delimit the surgical training field. The objective of the model is to simulate only the microsurgical step when the vas deferens are already isolated. The parameters analyzed were: feasibility for reproducing the technique, patency before and after performing the vasovasostomy, cost of the model, ease of acquisition, ease of handling, execution time, and model reproducibility. Results: The simulator presented low cost. All models made were viable with a texture similar to human, with positive patency obtained in 100% of the procedures. The internal and external diameters of the vas deferens varied between 0.2-0.4 mm and 2-3 mm, respectively, with a mean length of 9 ± 1.2 cm. The total procedure time was 43.28 ± 3.22 minutes. Conclusions: The realistic model presented proved to be viable for carrying out vasectomy reversal training, due to its low cost, easy acquisition, and easy handling, and providing similar tissue characteristics to humans.

Reproducibility of double agar gel immunodiffusion test using stored serum and plasma from paracoccidioidomycosis patients

Background: Serological evaluation performed by double agar gel immunodiffusion test (DID) is used for diagnosis, evaluation of severity, management of paracoccidioidomycosis patients, and development of new clinical studies. For these reasons, the Botucatu Medical School of UNESP maintains a serum bank at the Experimental Research Unit with patient clinical data. This study aimed to evaluate the influence of the freeze-thaw cycle and different blood matrices on the titration of circulating antibodies. Methods: The study included 207 patients with confirmed (etiology-demonstrated) or probable (serology-demonstrated) paracoccidioidomycosis, and DID was performed with culture filtrate from Paracoccidioides brasiliensis B339 as antigen. First experiment: the antibody levels were determined in serum samples from 160 patients with the chronic form and 20 with the acute/subacute form, stored at ­80o C for more than six months. Second experiment: titers of 81 samples of serum and plasma with ethylenediaminetetraacetic acid (EDTA) or heparin, from 27 patients, were compared according to matrix and effect of storage at ­20o C for up to six months. Differences of titers higher than one dilution were considered discordant. Results: First experiment: test and retest presented concordant results in serum stored for up to three years, and discordant titers in low incidence in storage for four to six years but high incidence when stored for more than six years, including conversion from reagent test to non-reagent retest. Second experiment: serum, plasma-EDTA and plasma-heparin samples showed concordant titers, presenting direct correlation, with no interference of storage for up to six months. Conclusions: Storage at ­80o C for up to six years has no or little influence on the serum titers determined by DID, permitting its safe use in studies depending on this parameter. The concordant titrations in different blood matrices demonstrated that the plasma can be used for immunodiffusion test in paracoccidioidomycosis, with stability for at least six months after storage at ­20o C.(AU)

Validation of a one-tube nested real-time PCR assay for the detection of Cryptosporidium spp. in avian fecal samples / Validação da nested PCR em tempo real em um tubo para detecção de Cryptosporidium spp. em amostras fecais de aves

Abstract The aim of this study was to validate a one-tube nested real-time PCR assay followed by genetic sequencing to detect and identify Cryptosporidium species and genotypes in birds. A total of 443 genomic DNA extracted from avian fecal samples were analyzed by one-tube nested real-time PCR and conventional nested PCR. By one-tube nested real-time PCR, 90/443 (20.3%) samples were positive for Cryptosporidium spp. In contrast, 36/443 (8.1%) samples were positive for Cryptosporidium spp. by conventional nested PCR. The analytical sensitivity test showed that one-tube nested real-time PCR detects approximately 0.5 oocyst (2 sporozoites) per reaction. An evaluation of analytical specificity did not reveal amplification of microorganisms that commonly present nonspecific amplification with primers used for the diagnosis of Cryptosporidium spp. The repeatability analysis showed the same result in 27 out of 30 samples (90%). As for the reproducibility of one-tube nested real-time PCR, 24 of the 30 samples examined (80%) showed the same result. All the 90 samples amplified by one-tube real-time nested PCR were successfully sequenced, leading to the identification of C. baileyi, C. galli, C. meleagridis, C. proventriculi, and Cryptosporidium avian genotype I. Genetic sequencing of conventional nested PCR amplicons was successful in 10/36 (27.8%) of positive samples.

Resumo O objetivo deste trabalho foi validar um protocolo de nested PCR em tempo real em um tubo (nPCR-TR-1T) seguida de sequenciamento genético para detectar e caracterizar as espécies e genótipos de Cryptosporidium em aves. Um total de 443 amostras de DNA genômico, extraído de amostras fecais de aves, foi analisado pela nPCR-TR-1T e pela nested PCR convencional. Pela nPCR-TR-1T, foi observada positividade para Cryptosporidium spp. de 20,3% (90/443), em contraste com a nested PCR convencional, que apresentou positividade de 8,1% (36/443). O teste de sensibilidade analítica mostrou que a nPCR-TR-1T detecta aproximadamente 0,5 oocisto (2 esporozoítos) por reação. A avaliação da especificidade analítica não revelou amplificação de microrganismos que comumente apresentam amplificação inespecífica com primers utilizados para o diagnóstico de Cryptosporidium spp. O cálculo da repetibilidade evidenciou o mesmo resultado em 27 de 30 amostras (90%). Em relação à reprodutibilidade da nPCR-TR-1T, foi observado o mesmo resultado em 80% (24/30) das amostras examinadas. Foi possível realizar o sequenciamento em todas as 90 amostras amplificadas pela nPCR-TR-1T, com identificação de C. baileyi, C. galli, C. meleagridis, C. proventriculi e Cryptosporidium genótipo I em aves. O sequenciamento dos fragmentos amplificados pela nested PCR convencional foi possível em 10/36 (27,8%) das amostras positivas.

Interobserver variability in the diagnosis of canine hepatoid gland tumors

The reported prevalence of canine hepatoid gland tumor (HGT) varies widely owing mainly to the lack of well-defined diagnostic criteria and poor interobserver agreement. The aim of the present study was to improve the level of diagnostic agreement among pathologists in canine HGT. Five pathologists diagnosed and classified morphological features in 57 cases of canine HGT and, based on their reports, diagnostic algorithms were devised using recursive partitioning analysis. The proportion of diagnoses of malignant hepatoid neoplasia among the five pathologists ranged from 26.3 to 50.9%. Interobserver diagnostic agreement was classified as fair (κ = 0.54) but improved to good (κ ~ 0.65) following application of two novel diagnostic algorithms based on histomorphological features as sebaceous differentiation, mitotic count, atypical mitosis and cellular atypia. This study has demonstrated that interobserver agreement in the diagnosis of canine HGT could be improved using novel algorithms. Further analyses are warranted to validate the proposed classification systems applying a higher sampling of canine HGTs.(AU)

Zebrafish (Danio rerio) meets bioethics: the 10Rs ethical principles in research / O peixe-zebra (Danio rerio) encontra a bioética: os princípios éticos dos 10Rs na pesquisa

Zebrafish (Danio rerio) is a tropical fish species widely used in research, worldwide. The development of genetically modified animals and the increasing number of zebrafish breeding facilities due to their emerging use in several research fields, opened room for new ethical challenges for research carried out with this species. It is necessary to raise the scientific community's awareness of the ethical standards and laws in force, on animal research. Thus, the aim of the current study is to describe 10 Rs ethical principles by using zebrafish as model system in research. The classical 3 Rs concerning animal welfare, namely replacement, reduction and refinement; and the added 7 Rs related to scientific (registration, reporting, robustness, reproducibility and relevance) and conduct principles (responsibility, and respect) in zebrafish research are herein presented and critically discussed. The use of these 10 Rs by researchers, institutions and the Animal Ethics Committee is recommended to support regulations, decision-making about and the promotion of zebrafish health and welfare in research.

O peixe-zebra (Danio rerio) é um peixe tropical amplamente usado em pesquisas em todo o mundo. Devido ao seu uso emergente em várias áreas de pesquisa, o desenvolvimento de animais geneticamente modificados e o aumento das instalações de peixe-zebra, novos desafos éticos surgem na pesquisa com o peixe-zebra. Além disso, é necessária a conscientização da comunidade científica sobre as normas éticas e leis vigentes na pesquisa com animais. Assim, o presente estudo teve como objetivo descrever os princípios éticos de 10 Rs usando o peixe-zebra como sistema modelo em pesquisa. Os 3 Rs clássicos relativos ao bem-estar animal (substituição, redução e refinamento) e 7 Rs adicionais relacionados aos princípios científicos (registro, relatório, robustez, reprodutibilidade e relevância) e de conduta (responsabilidade e respeito) na pesquisa do peixe-zebra são apresentados e discutido criticamente. Recomendamos o uso desses 10 Rs pelos pesquisadores, instituições e Comitê de Ética Animal na regulamentação, decisão e promoção da saúde e bem-estar do peixe-zebra em pesquisas.

Multiparametric assay of screening for the diagnosis of mycoses of interest in Public Health: standardization of methodology / Ensaio multiparamétrico de triagem para diagnóstico de micoses de interesse em Saúde Pública: padronização da metodologia

The standardization and validation of a multiplex assay requires the combination of important parameters such as sensitivity and specificity, acceptable levels of performance, robustness, and reproducibility. We standardized a multiparametric Dot-blot aimed at the serological screening of paracoccidioidomycosis, histoplasmosis, and aspergillosis. A total of 148 serum were evaluated: 10 from healthy subjects, 36 from patients with paracoccidioidomycosis, 62 from patients with histoplasmosis, and 40 from patients with aspergillosis. It was found that the multiparametric Dot-blot showed a high percentage of cross-reactivity. However, when evaluated individually, in the serological screening of histoplasmosis, a good performance was observed when compared to the double immunodiffusion assay, considered the gold standard test, with 100% co-positivity and 83.3% co-negativity. The performance of serological screening for aspergillosis was not satisfactory when compared to double immunodiffusion, showing 71.4% co-positivity and 100% co-negativity. The evaluation of the stability of nitrocellulose membranes showed that membranes sensitized with H. capsulatum antigen remained stable for 90 days and those sensitized with A. fumigatus antigen for 30 days. We conclude that the use of crude antigens was not suitable for the standardization of the multiparametric Dot-blot assay, due to the high cross-reactivity, and that further tests should be performed with purified proteins (AU).

A padronização e validação de um ensaio multiplex requer a combinação de parâmetros importantes, como sensibilidade e especificidade, níveis aceitáveis de desempenho, robustez e reprodutibilidade. Este trabalho padronizou um Dot-blot multiparamétrico visando a triagem sorológica da paracoccidioidomicose, histoplasmose e aspergilose. Foram avaliadas 148 amostras de soro: 10 de indivíduos saudáveis, 36 de pacientes com paracoccidioidomicose, 62 de pacientes com histoplasmose e 40 de pacientes com aspergilose. Verificou-se que o Dot-blot multiparamétrico apresentou elevado percentual de reatividade cruzada. Entretanto, quando avaliado individualmente, na triagem sorológica da histoplasmose observou-se bom desempenho quando comparado ao ensaio de imunodifusão dupla, considerado o teste padrão ouro, com 100% de co-positividade e 83,3% de co-negatividade. O desempenho da triagem sorológica da aspergilose não foi satisfatório quando comparado a imunodifusão dupla, apresentando 71,4% de co-positividade e 100% de co-negatividade. A avaliação da estabilidade das membranas de nitrocelulose mostrou que membranas sensibilizadas com antígeno de H. capsulatum permaneceram estáveis por 90 dias e as sensibilizadas com antígeno de A. fumigatus, por 30 dias. Concluímos que o uso de antígenos brutos não foi adequado para a padronização do ensaio de Dot-blot multiparamétrico, devido ao alto índice de reatividade cruzada, e que novos testes devem ser realizados com proteínas purificadas (AU).

Lateral and medial musculatures of pelvic limbs in dogs - normal ultrasonographic anatomy

Background: Pelvic limb claudication is a common problem in dogs. However, determining the location of the lesion can be challenging. Diagnostic imaging is essential for identifying and evaluating muscle injuries. Ultrasonographic evaluation is frequently performed to identify muscle injuries. Although some muscle changes have already been described as ultrasonography findings in veterinary studies, little is known about the standardization of these muscle structures, especially in hip muscles. The objective of this study was to describe the normal ultrasound anatomy of the lateral and medial muscles of the pelvic limb and establish an unprecedented protocol for the evaluation of these structures. Materials, Methods & Results: A total of 4 lateral and 4 medial musculatures of the pelvic limb of dogs were evaluated using ultrasound. An initial ex vivo study (1st phase) was performed to delineate anatomical and ultrasonographic assessments. The 2nd experimental phase proceeded with the application of in vivo scanning. The same operator performed all anatomical and ultrasonographic assessments. The gluteus medius, vastus lateralis, tensor fasciae latae, sartorius, pectineus, gracilis, adductor, and rectus femoris muscles were evaluated in longitudinal and transverse planes. Ten paired hips from 5 cadavers consisting of 3 females (60%) and 2 males (40%) were evaluated during the exvivo phase. The average age and weight of the dogs were 3.6 ± 0.87 years and 21.9 ± 5.2 kg, respectively. The invivo study included 4 females (40%) and 6 males (60%), for a total of 10 healthy animals consisting of 20 hips; the mean weight was 30.11 ± 7.45 kg and the average age was 4.5 ± 2.75 years. All muscles maintained the same ultrasound pattern when compared with each other and between different groups in the invivo study. The longitudinal section presented a hypoechoic structure permeated by thin hyperechoic longitudinal striations. The hypoechoic aspect was maintained, with striae identified as hyperechogenic points in the cross-sectional view. Discussion: B-mode ultrasonographic evaluation with a high-frequency transducer has facilitated the identification of the lateral and medial muscles of the pelvic limbs of medium and large dogs exvivo and invivo. This approach is an effective and safe way to access these muscles, and aids in the establishment of an unprecedented protocol for ultrasound evaluation of this region. Understanding the normal ultrasonographic anatomy is essential to identify muscle injuries such as contractures, tumors, ruptures, and stretch, which can lead to loss of muscle echogenicity and echotexture. Furthermore, ultrasound standardization reduces inter-observer error, facilitates technique reproducibility, assists in a more assertive assessment, and directs the patient's most appropriate therapy. In addition, anatomical dissection with the aid of acoustic gel injection into the evaluated musculature is essential to guide the examination because it allows the confirmation of the structures. In this study, the ultrasound scans of 6 fundamental muscles (gluteus medius, tensor fasciae latae, gracilis, adductor, pectineus, and rectus femoris) for hip mobility and 2 knee muscles (sartorius and vastus lateralis) were standardized. Exvivo evaluations facilitated the application of invivo techniques, and no difficulties were found during these evaluations.

Development and validation of a simulator for teaching minimally invasive thoracic surgery in Brazil

ABSTRACT Purpose To develop and validate a chest cavity simulator for teaching video-assited thoracic surgery (VATS). Methods The first phase of the study consisted of developing a chest cavity simulator. A quasi-experimental study was performed in the second phase, and 25 surgeons and residents participated in a three-stage pulmonary suture experiment. The videos were recorded and timed. Generalized linear regression models for repeated measures were used to analyze the outcome change over time. Results The chest cavity simulator consists of a console simulating the left hemithorax. Among the participants, 96% rated the design, visual aspect, positioning ergonomics, and triangulation of the portals as very good or excellent (face validity). There was a decrease in suturing time in step 1 from 435.7 ± 105 to 355.6 ± 76.8 seconds compared to step 3 (p = 0.001). The evaluation of the simulation effectiveness and performance (content validity) was rated as very good or excellent by 96% ofparticipants. The most experienced surgeon showed significant reduction in procedure time (p = 0.021) (construct validity). Conclusions The thoracic cavity simulator is realistic, showing content and construct validity, and can be used in VATS training. The simulation model allowed skill gain in the endoscopic suture.

Integrative taxonomy: combining molecular and morphological characteristics to identify Lymnaea (Galba) cubensis, intermediate host of Fasciola hepatica / Taxonomia integrativa: combinando características moleculares e morfológicas para identificar Lymnaea (Galba) cubensis, hospedeiro intermediário de Fasciola hepatica

Abstract Despite the epidemiological importance of the Lymnaeidae family regarding transmission of Fasciola hepatica, knowledge about the diversity and distribution of these molluscs and the role of each species in the expansion of fasciolosis remains sparse. Classical morphological (n=10) identification was performed in lymneids from Lagoa Santa, a municipality in the state of Minas Gerais, Brazil, along with molecular and phylogenetic analysis (n=05) based on the partial nucleotide sequences of the mitochondrial cytochrome c oxidase subunit I gene (COI mtDNA) and ribosomal internal transcribed spacer II (ITS-2 rDNA). The shell morphology made it possible to distinguish the lymneids of Lagoa Santa from Pseudosuccinea columella. Differences found in the penile complex and prostate shape allowed this species to be distinguished from Galba truncatula. However, the homogeneity of reproductive tract characteristics among Lymnaea (Galba) cubensis, L. viator and L. neotropica confirmed that these characteristics show low taxonomic reliability for identifying cryptic species. Genetic divergence analysis for the COI mtDNA gene and ITS-2 region of rDNA revealed greater similarity to Lymnaea (Galba) cubensis. Thus, correct species differentiation is important for monitoring the epidemiological risk of fasciolosis in the state of Minas Gerais, where cases of the disease have increased over recent years.

Resumo Apesar da importância epidemiológica da família Lymnaeidae na transmissão de Fasciola hepatica, o conhecimento sobre a diversidade e a distribuição desses moluscos e o papel de cada espécie, na expansão da fasciolose, ainda é escasso. Realizou-se a identificação morfológica clássica (n=10) em limneídeos de Lagoa Santa, município do estado de Minas Gerais, Brasil, juntamente com a análise molecular e filogenética (n=05), baseada nas sequências parciais de nucleotídeos do gene mitocondrial da subunidade I do citocromo c oxidase (COI mtDNA) e espaçador interno, transcrito do DNA ribossomal II (ITS-2 rDNA). A morfologia da concha possibilitou distinguir os limneídeos de Lagoa Santa de Pseudosuccinea columella. As diferenças encontradas no complexo peniano e na forma da próstata permitiram que essa espécie fosse distinta de Galba truncatula. No entanto, a homogeneidade das características do trato reprodutivo entre Lymnaea (Galba) cubensis, L. viator e L. neotropica confirmou que essas características apresentam baixa confiabilidade taxonômica para a identificação de espécies crípticas. A análise da divergência genética para o gene COI mtDNA e região ITS-2 do rDNA revelou maior similaridade entre os limneídeos de Lagoa Santa com Lymnaea (Galba) cubensis.

Diagrammatic scales for the estimation of black node disease severity in common bean / Escalas diagramáticas para a estimativa da gravidade da doença do nó negro no feijão comum

The objective of this study was to design two diagrammatic scales for the evaluation of black node disease (Stagonosporopsis hortensis and Boeremia spp.) in common bean (Phaseolus vulgaris L.). The developed scales corresponded to logarithmic intervals (SADL) of seven levels and equal intervals (SADE) of nine levels. These scales were compared to the scale developed by the International Center for Tropical Agriculture (CIAT), which corresponds to a descriptive scale (NoSAD). In the scale validation the accuracy, precision, and concordance of repeatability were determined using Lins concordance correlation coefficient, whereas inter-rater reproducibility was determined by the overall concordance correlation coefficient (OCCC). It was observed that with the use of the proposed scales, reproducibility and repeatability were significantly improved for inexperienced raters, and reproducibility was improved in the case of experienced raters. Thus, the designed standard area diagrams with equal and logarithmic intervals are a useful tool for estimating severity under field and experimental conditions as part of the study of this patho-system.(AU)

O objetivo deste estudo foi delinear duas escalas diagramáticas para avaliação da doença do nó preto (Stagonosporopsis hortensis e Boeremia spp.) em feijoeiro (Phaseolus vulgaris L.). As escalas desenvolvidas corresponderam a intervalos logarítmicos (SADL) de sete níveis e intervalos iguais (SADE) de nove níveis. Essas escalas foram comparadas com a escala desenvolvida pelo Centro Internacional de Agricultura Tropical (CIAT), que corresponde a uma escala descritiva (NoSAD). Na validação da escala, a exatidão, precisão e concordância da repetibilidade foram determinadas usando o coeficiente de correlação de concordância de Lin, enquanto a reprodutibilidade entre avaliadores foi determinada pelo coeficiente de correlação de concordância geral (OCCC). Observou-se que, com o uso das escalas propostas, a reprodutibilidade e a repetibilidade foram significativamente melhoradas para os avaliadores inexperientes, e a reprodutibilidade foi melhorada no caso dos avaliadores experientes. Assim, as escalas diagramáticas projetadas são uma ferramenta útil para estimar a severidade em condições experimentais e de campo como parte do estudo desse sistema patológico.(AU)

Standardization of blood meal production process at a rendering plant using design of experiments / Padronização do processo de produção de farinha de sangue em uma usina de processamento por meio do planejamento de experimentos

This research focuses on the standardization of the blood meal production process at a Colombian rendering plant through a design of experiments. Initially, 108 samples of blood meal were taken where only 23% achieved the moisture target (7.5% to 8.5%). Therefore, an analysis of the measurement system was performed using a repeatability and reproducibility (R&R) study. Results showed that 39.96% of the observed variability was caused by the measurement system that was out of control. So, it was necessary to improve the method of sampling reducing the participation of the measurement system in the variability of the process to only 3.79%. Later, several experiments were accomplished with a 2k factorial design. Each experiment consisted of a response variable (blood meal moisture), two controllable factors (drying chamber temperature and percentage of rotation of the metering screw), and an uncontrollable factor (initial blood meal moisture). Finally, experiments were carried out and validated observing that, with a drying chamber temperature of 160 °C and a percentage of screw rotation of 29%, more than 97% of the blood meal was according to the moisture target. In conclusion, is confirmed that the design of experiments is a tool that allows a clear path towards optimization and standardization of processes.(AU)

Esta pesquisa se concentra na padronização do processo de produção de farinha de sangue de uma usina de beneficiamento colombiana por meio de um planejamento de experimentos. Em primeiro lugar, foi realizado um estudo de 108 amostras de farinha de sangue onde apenas 23% atingiram a meta de umidade (7.5% a 8.5%). Portanto, uma análise do sistema de medição foi realizada usando um estudo de repetibilidade e reprodutibilidade (R&R). Os resultados mostraram que 39.96% da variabilidade observada foi causada pelo sistema. Assim, foi necessário aprimorar o método de amostragem, reduzindo a participação do sistema na variabilidade para apenas 3.79%. Posteriormente, vários experimentos foram realizados com um planejamento fatorial 2k. Cada experimento consistiu em uma variável de resposta (umidade da farinha de sangue), dois fatores controláveis (temperatura da câmara de secagem e porcentagem de rotação da rosca dosadora) e um fator incontrolável (umidade inicial da farinha de sangue). Finalmente, experimentos foram realizados e validados observando que, com uma temperatura da câmara de secagem de 160 ° C e um percentual de rotação da rosca de 29%, mais de 97% da farinha de sangue estava de acordo com a umidade desejada. Em conclusão, confirma-se que o planejamento de experimentos é uma ferramenta que permite um caminho claro para a otimização e padronização de processos.(AU)

Validation of Alternative Methods of Detection of Salmonella Spp. in Experimentally Contaminated Poultry Environmental Samples

The analysis of Salmonella in the feces and the birds environment is a way of monitoring the colonization in the flocks and verifying the need for the introduction of stricter controls, in such a way that the results of the tests should be known before being sent for slaughter. The polymerase chain reaction (PCR), as well as other rapid methods represent alternatives increasingly used to detect enteric pathogens, but they need proof of effectiveness for their wide use. The aim of this study was to evaluate the equivalence between the results obtained by the methods: real-time PCR (BAX® System), Modified Rappaport-Vassiliadis Semi-solid Medium (MSRV) (ISO 6579) and the traditional method of official reference in Brazil for research of S. Typhimurium and S. Enteritidis in poultry samples. Two hundred and fifty-two samples of disposable shoe covers (DSC) and 252 samples of feces were infected with an average of 2 to 3 log CFU/g of each serovar, and the same samples without fortification were evaluated by the three methods. Five hundred and four diagnoses were obtained with satisfactory results in terms of repeatability (greater than 80%), reproducibility (mean 83,1%), sensitivity (81% to 100%), specificity (95% to 100%), and accuracy (90% to 100%). The compliance test verified that there was not a significant difference between the alternative and the official methods, allowing us to state that the methodologies have had equivalent performances.(AU)

Validation of Alternative Methods of Detection of Salmonella Spp. in Experimentally Contaminated Poultry Environmental Samples

The analysis of Salmonella in the feces and the birds environment is a way of monitoring the colonization in the flocks and verifying the need for the introduction of stricter controls, in such a way that the results of the tests should be known before being sent for slaughter. The polymerase chain reaction (PCR), as well as other rapid methods represent alternatives increasingly used to detect enteric pathogens, but they need proof of effectiveness for their wide use. The aim of this study was to evaluate the equivalence between the results obtained by the methods: real-time PCR (BAX® System), Modified Rappaport-Vassiliadis Semi-solid Medium (MSRV) (ISO 6579) and the traditional method of official reference in Brazil for research of S. Typhimurium and S. Enteritidis in poultry samples. Two hundred and fifty-two samples of disposable shoe covers (DSC) and 252 samples of feces were infected with an average of 2 to 3 log CFU/g of each serovar, and the same samples without fortification were evaluated by the three methods. Five hundred and four diagnoses were obtained with satisfactory results in terms of repeatability (greater than 80%), reproducibility (mean 83,1%), sensitivity (81% to 100%), specificity (95% to 100%), and accuracy (90% to 100%). The compliance test verified that there was not a significant difference between the alternative and the official methods, allowing us to state that the methodologies have had equivalent performances.

Canal flare index evaluation for different dog breeds / Avaliação do canal flare index femoral de diferentes raças de cães

This study determined the canal flare index (CFI) of four dog breeds using two distinct femoral regions as a reference. Thirty-five radiographs of the hip joints of Golden Retrievers (GRG), German Shepherds (GSG), Labrador Retrievers (LRG), and Rottweilers (RG) of both sexes were used. Seventy experimental units were submitted to CFI calculation. Objective (CFIob) and subjective (CFIsub) values of the CFI of each experimental unit were determined according to the anatomical reference used for the calculation. A significant difference in the CFIob between the Golden Retriever and German Shepherd breeds (1.68 ± 0.16 and 1.49 ± 0.08), and in the CFIsub between Golden Retriever, German Shepherd, and Rottweiler breeds (2.09 ± 0.31, 1.86 ± 0.11, and 1.84 ± 0.18) was reported. The subjective form of measurement showed higher values than the objective form (GRG: 2.09 ± 0.31; GSG: 1.86 ± 0.11; LRG: 2.07 ± 0.12; RG: 1.84 ± 0.18). The CFI values of each breed were similar, suggesting a certain racial pattern. A significant difference in the interobserver assessment for both CFIsub and CFIob, in all races was observed. The CFI analysis identified morphological patterns of the proximal femur in the different races. Results indicated the need for standardization of the anatomical references used to calculate the CFI because there were statistical differences among the measurements among the observers.(AU)

Objetivou-se determinar o canal flare index (CFI) de quatro raças específicas de cães, utilizando-se duas regiões femorais distintas como referência. Foram analisadas 35 radiografias de articulações coxofemorais de cães das raças Golden Retriever (GGR), Pastor Alemão (GPA), Labrador Retriever (GLR) e Rottweiler (GR), de ambos os sexos, configurando 70 unidades experimentais submetidas ao cálculo do CFI. Determinaram-se os valores objetivo (CFIob) e subjetivo (CFIsub) do CFI de cada unidade experimental de acordo com a referência anatômica utilizada para o cálculo. Houve diferença significativa do CFIob entre as raças Golden Retriever e Pastor Alemão (1,68 ± 0,16 e 1,49 ± 0,08), e do CFIsub entre as raças Golden Retriever, Pastor Alemão e Rottweiler (2,09 ± 0,31, 1,86 ± 0,11 e 1,84 ± 0,18). A forma subjetiva de mensuração apresentou valores maiores que a forma objetiva (GGR: 2,09 ± 0,31; GPA: 1,86 ± 0,11; GLR: 2,07 ± 0,12; GR: 1,84 ± 0,18). Os valores de CFI de cada raça se apresentaram similares, sugerindo haver determinado padrão racial. Houve diferença significativa na avaliação interobservadores, tanto do CFIsub quanto CFIob, em todas as raças. A análise do CFI identificou padrões morfológicos do fêmur proximal nas diferentes raças. Os resultados indicaram a necessidade de padronização dos referenciais anatômicos utilizados para o cálculo do CFI, uma vez que houve relevância estatística entre as diferentes mensurações entre os observadores.(AU)

A standard area diagram set for assessing severity of eucalyptus bacterial blight caused by Erwinia psidii / Uma escala diagramática para a avaliação da severidade da seca-de-ponteiros do eucalipto causada por Erwinia psidii

This study aimed to develop and validate a standard area diagram (SAD) set to estimate the severity of bacterial blight of eucalyptus caused by Erwinia psidii. For this purpose, an eight-level SAD was developed and validated by ten inexperienced raters. Accuracy and precision of the estimates by each rater, with and without the SAD, were determined based on Lin's concordance correlation coefficient. The proposed SAD improved the accuracy and precision of the estimates. The SAD set studied here is a useful tool in assessments of bacterial blight of eucalyptus for epidemiological research and breeding programs.(AU)

Este trabalho objetivou o desenvolvimento de uma escala para estimar a severidade da seca-de-ponteiros do eucalipto causada por Erwinia psidii. Para isso, uma escala de oito níveis foi desenvolvida e validada por dez avaliadores inexperientes. A acurácia e precisão das estimativas de cada avaliador, com e sem a escala, foram determinadas baseadas no coeficiente de correlação concordante de Lin. A escala proposta melhorou a acurácia e a precisão das estimativas. A escala estudada se mostrou uma ferramenta útil na avaliação da seca-de-ponteiros do eucalipto para estudos epidemiológicos e em programas de melhoramento.(AU)

A standard area diagram set for assessing severity of eucalyptus bacterial blight caused by Erwinia psidii / Uma escala diagramática para a avaliação da severidade da seca-de-ponteiros do eucalipto causada por Erwinia psidii

This study aimed to develop and validate a standard area diagram (SAD) set to estimate the severity of bacterial blight of eucalyptus caused by Erwinia psidii. For this purpose, an eight-level SAD was developed and validated by ten inexperienced raters. Accuracy and precision of the estimates by each rater, with and without the SAD, were determined based on Lin's concordance correlation coefficient. The proposed SAD improved the accuracy and precision of the estimates. The SAD set studied here is a useful tool in assessments of bacterial blight of eucalyptus for epidemiological research and breeding programs.(AU)

Este trabalho objetivou o desenvolvimento de uma escala para estimar a severidade da seca-de-ponteiros do eucalipto causada por Erwinia psidii. Para isso, uma escala de oito níveis foi desenvolvida e validada por dez avaliadores inexperientes. A acurácia e precisão das estimativas de cada avaliador, com e sem a escala, foram determinadas baseadas no coeficiente de correlação concordante de Lin. A escala proposta melhorou a acurácia e a precisão das estimativas. A escala estudada se mostrou uma ferramenta útil na avaliação da seca-de-ponteiros do eucalipto para estudos epidemiológicos e em programas de melhoramento.(AU)

Standard area diagram set for scab quantification in trilobed leaves of the sour passion fruit / Escala diagramática para a quantificação da verrugose em folhas trilobadas do maracujazeiro azedo

The study aimed to develop and validate a standard area diagram set (SADs) to improve accuracy, precision, and reproducibility in the assessment of scab severity in trilobed leaves of the sour passion fruit. The SADs comprised six severity levels (1%; 3%; 6%; 11%; 21%; and 33%). The SADs increased the accuracy of the estimates by reducing the constant and systematic errors, and the tendency to overestimate the actual severity, limited the number of evaluators who presented this trend, in addition to increasing the bias correction factor. The inexperienced evaluators were the ones that benefited the most from the SADs use. Once the SADs was introduced, 80% and 100% evaluators gave estimates free from constant and systematic errors, respectively. The improvement in precision was verified by the increase in the coefficient of determination and Lins concordance correlation coefficient, by reducing absolute errors and increasing the reproducibility of the estimates.(AU)

O objetivo deste estudo foi desenvolver e validar uma escala diagramática para melhorar a acurácia, precisão e reprodutibilidade na avaliação da severidade da verrugose em folhas trilobadas do maracujazeiro azedo. A escala apresentou seis níveis de severidade (1; 3; 6; 11; 21 e 33%) e aumentou a acurácia das estimativas, reduzindo os erros constantes e sistemáticos, a tendência de superestimação em relação aos valores de severidade real e o número de avaliadores que apresentavam essa tendência, além de aumentar o fator de correção do desvio. Os avaliadores inexperientes foram os mais beneficiados pela utilização da escala, sendo que 80% e 100% destes avaliadores apresentaram estimativas livres de erros constantes e sistemáticos, respectivamente, quando a escala foi empregada. O aumento da precisão foi verificado pelo incremento dos coeficientes de determinação e de concordância de Lin, redução dos erros absolutos e aumento da reprodutibilidade das estimativas.(AU)

Reprodutibilidade e repetibilidade de mensuração radiográfica da silhueta cardíaca em cães da raça poodle / Reproducibility and repeatability of radiographic measurement of the cardiac silhouette in poodles

Avaliou-se o potencial de reprodutibilidade e repetibilidade de um sistema semiautomático de mensuração radiográfica. Para isto foram mensurados, por intermédio de um software o vertebral heart score (VHS) e a área cardíaca normalizada (ACN) em imagens radiográficas de cães da raça poodle clinicamente normais e portadores de insuficiência valvar mitral. Dada a subjetividade da análise da silhueta cardíaca por diferentes observadores, acredita-se que mesmo com um sistema semiautomático essa avaliação poderia ser afetada pela experiência prévia do observador em radiodiagnóstico. Foram analisadas cinquenta e sete imagens radiográficas torácicas de cães atendidos em um hospital veterinário. As imagens foram analisadas por dois grupos de observadores com diferentes níveis de experiência em radiologia: (A) sem experiência (n = 2) e (B) com experiência (n = 2). Demonstrou-se boa repetibilidade e reprodutibilidade para o VHS independente da experiência em radiodiagnóstico, e para a ACN em observadores experientes e com familiaridade com o software.

The potential of reproducibility and repeatability of a semi-automatic system of radiographic measurement was evaluated. For this purpose, the vertebral heart score (VHS) and normalized cardiac area (NCA) were evaluated in radiographs of healthy poodles and of patients with mitral regurgitation. Considering the subjective aspect of the cardiac silhouette analysis by different observers, it is believed that even with a semi-automatic system this evaluation could be affected by the previous experience of the observer in radiodiagnosis. Fifty seven thoracic images were analyzed by two groups of observers with different levels of experience in radiology: (A) without experience (n = 2) and (B) with experience (n = 2). Regardless of radiodiagnostic experience it was observed good repeatability and reproducibility for VHS, and also for ACN in observers with experience in radiodiagnostics and previous familiarity with the software.

Reprodutibilidade e repetibilidade de mensuração radiográfica da silhueta cardíaca em cães da raça poodle / Reproducibility and repeatability of radiographic measurement of the cardiac silhouette in poodles

Avaliou-se o potencial de reprodutibilidade e repetibilidade de um sistema semiautomático de mensuração radiográfica. Para isto foram mensurados, por intermédio de um software o vertebral heart score (VHS) e a área cardíaca normalizada (ACN) em imagens radiográficas de cães da raça poodle clinicamente normais e portadores de insuficiência valvar mitral. Dada a subjetividade da análise da silhueta cardíaca por diferentes observadores, acredita-se que mesmo com um sistema semiautomático essa avaliação poderia ser afetada pela experiência prévia do observador em radiodiagnóstico. Foram analisadas cinquenta e sete imagens radiográficas torácicas de cães atendidos em um hospital veterinário. As imagens foram analisadas por dois grupos de observadores com diferentes níveis de experiência em radiologia: (A) sem experiência (n = 2) e (B) com experiência (n = 2). Demonstrou-se boa repetibilidade e reprodutibilidade para o VHS independente da experiência em radiodiagnóstico, e para a ACN em observadores experientes e com familiaridade com o software.(AU)

The potential of reproducibility and repeatability of a semi-automatic system of radiographic measurement was evaluated. For this purpose, the vertebral heart score (VHS) and normalized cardiac area (NCA) were evaluated in radiographs of healthy poodles and of patients with mitral regurgitation. Considering the subjective aspect of the cardiac silhouette analysis by different observers, it is believed that even with a semi-automatic system this evaluation could be affected by the previous experience of the observer in radiodiagnosis. Fifty seven thoracic images were analyzed by two groups of observers with different levels of experience in radiology: (A) without experience (n = 2) and (B) with experience (n = 2). Regardless of radiodiagnostic experience it was observed good repeatability and reproducibility for VHS, and also for ACN in observers with experience in radiodiagnostics and previous familiarity with the software.(AU)

Use of computed radiology as a screening test for the identification of congenital heart disease in dogs / Utilização da radiologia computadorizada como exame de triagem na identificação de cardiopatias congênitas em cães

The objectives of this study were to evaluate the accuracy of thoracic radiology as a screening test for congenital heart diseases in dogs, to identify the main contributions and limitations of this modality, and to verify the reproducibility of the evaluations by three observers with different levels of training. An interobserver, observational, retrospective and prospective study was carried out, including ninety dogs: thirty healthy animals, thirty with acquired heart diseases and thirty with congenital heart diseases, which all had thoracic radiographs and a confirmed echocardiographic diagnosis. The cases were separated and randomized by a mediator who did not participate in the reading of the radiographic examinations, and no evaluator had access to the patients' data. Sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV) and accuracy of each observer were calculated in relation to the correct classification of dogs to groups of normal or acquired and congenital heart diseases, as well as identification of enlargement of the cardiac silhouette and large vessels of dogs with congenital heart diseases. Finally, the Kappa coefficient was obtained between the observers to verify the reproducibility of the radiological evaluations performed. In general, sensitivity, PPV and accuracy were unsatisfactory ( 70%), and the agreement ranged from poor to reasonable (between 0 and 0.39). Although greater accuracy was achieved in the differentiation of healthy dogs from those with acquired and congenital heart diseases by thoracic radiography, when compared to the other studies, the modality was able only to identify healthy patients, and could not differentiate the individuals with different forms of heart disease or define the cardiac malformations. In addition, there was low reproducibility between observers, therefore, this technique should not be used as a sole screening method in dogs with suspected congenital heart diseases.(AU)

Os objetivos do estudo foram avaliar a acurácia da técnica radiográfica de tórax como exame de triagem nas cardiopatias congênitas em cães, identificar as principais contribuições e limitações dessa modalidade, e verificar a reprodutibilidade das avaliações realizadas por três observadores com diferentes graus de treinamento. Realizou-se um estudo interobservador, observacional, retrospectivo e prospectivo, o qual foram selecionados 90 cães, sendo 30 saudáveis, 30 com cardiopatias adquiridas e 30 com cardiopatias congênitas, que possuíam radiografia de tórax e diagnóstico ecocardiográfico confirmado. Os casos foram separados e randomizados por um mediador que não participou da leitura dos exames radiográficos, sendo que nenhum avaliador obteve acesso aos dados dos pacientes. Calculou-se os índices de sensibilidade, especificidade, valor preditivo positivo (VPP), valor preditivo negativo (VPN) e acurácia de cada observador em relação a identificação dos pacientes saudáveis, com cardiopatias adquiridas e congênitas, bem como para identificação de aumento da silhueta cardíaca e região de grandes vasos dos cães com cardiopatias congênitas. Por fim, foi obtido o coeficiente de Kappa entre os observadores a fim de verificar a reprodutibilidade das avaliações radiográficas realizadas. De modo geral, a sensibilidade, VPP e acurácia foram insatisfatórios (< 70%), enquanto a especificidade e VPN foram satisfatórios (> 70%), sendo que a concordância variou de ruim a razoável (entre 0 e 0,39). Apesar de ter sido alcançado acurácia maior na identificação de cães saudáveis, com cardiopatias adquiridas e congênitas pela radiografia de tórax quando comparado aos demais estudos, confirma-se que essa modalidade se mostrou capaz apenas de identificar o paciente saudável, não de diferenciar os indivíduos cardiopatas entre si ou de definir com maior detalhe as malformações cardíacas especificamente. Além disso, o exame radiográfico de tórax apresentou baixa reprodutibilidade entre os observadores, portanto, essa técnica não deve ser considerada como método único de triagem na suspeita de cães com cardiopatias congênitas.(AU)