Ocorrência de sorotipos exóticos de Salmonella encontrados em cães assintomáticos nos distritos do município de Ilhéus / BA - Brasil

Fecal samples from dogs residing in areas of poor living conditions in Ilhéus city / BA were examined for Salmonella spp from June 2001 to April 2002. Eighteen (9.47%) of 190 animals were found to be positive. 66.6% of those samples were taken from one year old puppies, 56% of the positive animals were fed by table scrap, 66% did not drink treated water and 67% had a domiciliary behavior. Symptoms of salmonellosis were not present in 83% of all positive animals. S. Gafsa represented the most prevalent serovar (38.9%), followed by S. Rubislaw (27.8%), S. Carrau (16.7%) and S. enterica subsp. Houtenae (11.1%). One sample could not be identified. Gentamicin was the most potent in vitro drug for the treatment of salmonelosis, followed by ampicilin. Our results confirmed that dogs represent an important reservoir of exotic Salmonella serovars and that there are different patterns of sensitivity to drugs among them.Amostras fecais de cães provenientes de distritos carentes do Município de Ilhéus / BA foram analisadas para a presença de Salmonella spp durante o período de junho de 2001 a abril de 2002. Dezoito (9,47%) dos 190 animais foram considerados positivos sendo que 66,6% destes casos ocorreram com animais até 1 ano de idade, 56% se alimentavam com comida caseira, 66% tinham acesso à fonte de água não tratada e 67% possuía comportamento domiciliar. A sintomatologia da doença não foi percebida em 83,3% dos animais. A S. Gafsa representou o sorotipo mais prevalente (38,9%), seguido pela S. Rubislaw (27,8), S. Carrau (16,7%) e S. enterica subsp. Houtenae (11,1%). Uma cepa não pôde ser identificada. O antibiograma revelou a gentamicina como sendo a droga mais potente, in vitro, para o tratamento das salmonelas encontradas na região de Ilhéus, seguida pela ampicilina. Nossos resultados confirmam que cães representam um importante reservatório de sorotipos exóticos de Salmonella e que ocorre padrões diferentes de sensibilidade a antibióticos entre eles

Panoramic snapshot of serum soluble mediator interplay in pregnant women with convalescent COVID-19: an exploratory study

IntroductionSARS-CoV-2 infection during pregnancy can induce changes in the maternal immune response, with effects on pregnancy outcome and offspring. This is a cross-sectional observational study designed to characterize the immunological status of pregnant women with convalescent COVID-19 at distinct pregnancy trimesters. The study focused on providing a clear snapshot of the interplay among serum soluble mediators.MethodsA sample of 141 pregnant women from all prenatal periods (1st, 2nd and 3rd trimesters) comprised patients with convalescent SARS-CoV-2 infection at 3-20 weeks after symptoms onset (COVID, n=89) and a control group of pre-pandemic non-infected pregnant women (HC, n=52). Chemokine, pro-inflammatory/regulatory cytokine and growth factor levels were quantified by a high-throughput microbeads array.ResultsIn the HC group, most serum soluble mediators progressively decreased towards the 2nd and 3rd trimesters of pregnancy, while higher chemokine, cytokine and growth factor levels were observed in the COVID patient group. Serum soluble mediator signatures and heatmap analysis pointed out that the major increase observed in the COVID group related to pro-inflammatory cytokines (IL-6, TNF-α, IL-12, IFN-γ and IL-17). A larger set of biomarkers displayed an increased COVID/HC ratio towards the 2nd (3x increase) and the 3rd (3x to 15x increase) trimesters. Integrative network analysis demonstrated that HC pregnancy evolves with decreasing connectivity between pairs of serum soluble mediators towards the 3rd trimester. Although the COVID group exhibited a similar profile, the number of connections was remarkably lower throughout the pregnancy. Meanwhile, IL-1Ra, IL-10 and GM-CSF presented a preserved number of correlations (≥5 strong correlations in HC and COVID), IL-17, FGF-basic and VEGF lost connectivity throughout the pregnancy. IL-6 and CXCL8 were included in a set of acquired attributes, named COVID-selective (≥5 strong correlations in COVID and <5 in HC) observed at the 3rd pregnancy trimester.Discussion and conclusionFrom an overall perspective, a pronounced increase in serum levels of soluble mediators with decreased network interplay between them demonstrated an imbalanced immune response in convalescent COVID-19 infection during pregnancy that may contribute to the management of, or indeed recovery from, late complications in the post-symptomatic phase of the SARS-CoV-2 infection in pregnant women

Diagnóstico da infecção por amebídeos através de técnicas morfológicas, imunológicas e moleculares em microrregião de Ilhéus - Itabuna, Bahia, Brasil

The Entamoeba gender possesses several species, hindering the diagnosis of amebiasis for morphologic likeness among protozoa, what carts in erroneous diagnosis and unnecessary treatment of many patients. Because of this, new techniques are being developed with the objective of differentiate those species. The present work has as objectives to evaluate the hygienic and sanitary conditions and the frequency of intestinal parasitosis on the studied population, identify and differentiate through immunological and molecular techniques the species of Entamoeba histolytica/ Entamoeba dispar complex and identify through morphological techniques the specie of Entamoeba hartmanni in residents of a micro region of Ilhéus-Itabuna. Thirty eight families participated in the study, having been collected 97 people's fecal samples. In the parasitological examination of feces, were used direct methods (Paratest) and the sedimentation method for centrifugation (modified Ritchie). The coproantigen research was accomplished through the commercial imunnoenzimactic method (ELISA, E. histolytica II-TECHLAB Inc. Blacksburg, VA, USA). In differentiation of species of E.histolytica/ E. dispar complex, the polymerase chain reaction of Multiplex PCR was accomplished and the E. hartmanni specie was identified through morphometry on slides stained by ferric hematoxylin. The results are suggestive that hygienic and sanitary conditions at the settlement are not satisfactory, being observed through the parasitological examination of feces a global frequency of enteroparasitosis by the order of 65%. The evaluation of specific frequency shown Ascaris lumbricoides (29%); Trichuris trichiura (10%); Ancylostomids (8%); Strongyloides stercoralis (3%), Entamoeba coli (25%); Endolimax nana (25%); Blastocistis hominis (13%); Entamoeba histolytica/Entamoeba dispar (25%); Giardia intestinalis (3%); Iodameba butschlii (1%). The method of floating-centrifugation was most efficient than the direct method on parasites recovering. The coproantigen research was efficient on identification of Entamoeba histolytica, showing that E. dispar was the most frequent specie in the studied population. The method of ferric hematoxylin colour was an important tool to identify E. hartmanni. This study shows that through association of various techniques, it is possible to identify amoebiasis agent, differentiating it from the other enteric amoebas, contributing to epidemiologic studies and avoiding unnecessary treatment of patients infected by other amoebas than E. histolyticaO gênero Entamoeba possui várias espécies, dificultando o diagnóstico da amebíase pela semelhança morfológica entre os protozoários, o que acarreta em diagnósticos errôneos e tratamento desnecessário de muitos pacientes. Por isso várias técnicas têm sido desenvolvidas, com o objetivo de diferenciar essas espécies. O presente trabalho teve como objetivos avaliar as condições higiênico-sanitárias e a freqüência de parasitos intestinais em moradores de uma microrregião de Ilhéus Itabuna, enfatizando a identificação e diferenciação através de técnicas imunológicas e moleculares as espécies do complexo Entamoeba histolytica/Entamoeba dispar e através de técnicas morfológicas a espécie Entamoeba hartmanni. De um total de 38 famílias, participaram do estudo 97 pessoas. No exame parasitológico das fezes cedidas pela população estudada, foram utilizados o métodos direto (Paratest) e o método de sedimentação por centrifugação (Ritchie modificado). A pesquisa de coproantígenos foi realizada através do método imunoenzimático comercial (ELISA, E. histolytica II TECHLAB Inc. Blacksburg, VA, USA). Na diferenciação das espécies do complexo Entamoeba histolytica/Entamoeba dispar, foi realizada a reação em cadeia da polimerase (PCR) - Multiplex e Entamoeba harmanni foi identificada através da morfometria em lâminas coradas por hematoxilina férrica. Os resultados sugerem que as condições higiênico-sanitárias no assentamento não são satisfatórias, tendo sido verificada através do exame parasitológico de fezes, uma frequência global de enteroparasitos de 65%. Na avaliação da frequência específica observou-se: Ascaris lumbricoides (29%), Trichuris trichiura (10%), Ancilostomídeo (8%), Strongyloides stercoralis (3%), Entamoeba coli (25%), Endolimax nana (25%), Blastocistis hominis (13%), Entamoeba histolytica/Entamoeba dispar (25%), Giardia intestinalis (3%), Iodameba butschlii (1%). O método de centrífugo-sedimentação foi mais eficaz que o direto na recuperação dos parasitos e a pesquisa de coproantígenos foi eficaz na identificação de Entamoeba histolytica, mostrando que E. dispar foi a espécie que predominou na população. O método de coloração hematoxilina férrica foi uma importante ferramenta para identificar E. hartmanni. Esse estudo mostrou que através da associação de diferentes técnicas é possível identificar o agente da amebíase, diferenciando-o das outras amebas intestinais, contribuindo para estudos epidemiológicos e evitando tratamento desnecessário de pacientes parasitados por outras amebas que não E. histolytic

Frequency of amoebiasis and other intestinal parasitoses in a settlement in Ilhéus City, State of Bahia, Brazil

Introduction: This study evaluated the frequency of intestinal parasites, emphasizing the identification and differentiation of Entamoeba spp. Methods: Multiplex polymerase chain reaction (PCR), coproantigen tests and morphometric analysis were performed for Entamoeba spp. differentiation. Results: The overall frequency of intestinal parasites was 65%. Entamoeba histolytica was detected by the coproantigen test, and the PCR showed that Entamoeba dispar predominated in the population. In contrast, morphometric analysis was important for identifying Entamoeba hartmanni. Conclusions: It is possible to identify the causative agent of amoebiasis and to differentiate this agent from other species by combining techniques

Monocytes and plasma tissue factor levels in normal individuals and patients with deep venous thrombosis of the lower limbs: potential diagnostic tools?

INTRODUCTION: Tissue factor (TF) is the main physiological initiator of blood coagulation; it is membrane-bound on monocytes (mTF) and free in plasma (pTF). Abnormal expression of TF by monocytes has been implicated in various diseases. We therefore quantified monocytes expressing TF and pTF levels in patients with lower-limb deep venous thrombosis (DVT).MATERIALS AND METHODS: DVT was confirmed by Duplex Scan. Blood mTF levels under resting condition (baseline), after incubation without (unstimulated) and with (stimulated) lipopolysaccharide (LPS), and total mTF levels were determined by flow cytometry using two analytical methods (Histogram and Quadrant-Statistics). Plasma TF levels were measured using an enzyme-linked immunoabsorbent assay (ELISA). Results were compared with age-matched controls.RESULTS: Histogram analysis in patients with DVT showed significantly elevated mTF levels for baseline, unstimulated and total mTF over controls. For Quadrant-Statistics, DVT patients also showed significantly raised baseline, unstimulated, stimulated and total mTF. Similarly, pTF levels were significantly raised in subjects with DVT compared to controls. Baseline mTF levels correlated with pTF levels by Histogram and Quadrant-Statistics analysis. Using the relative operating characteristic (ROC) curve, baseline mTF and pTF assays displayed sensitivity and specificity in detecting DVT. Quadrant-Statistics baseline mTF and pTF gave the best discrimination.CONCLUSIONS: The TF assays used in this study showed acceptable sensitivity and specificity and are cost-effective and practical. Therefore, they should be considered in patients with, or at risk of, DVT

Cryptococcosis in a Cat

Background: Cryptococcosis is a mycosis that primarily affects domestic cats and is caused by fungi of the genus Cryptococcus, resulting in cutaneous, ocular, respiratory, and neurological manifestations. Diagnosis is based on the microscopic evaluation of the lesions and isolation of the causative agent. The aim of this study was to describe a case of feline cryptococcosis diagnosed by cytology and treated at the Veterinary Hospital of the State University of Santa Cruz (HV-UESC). Case:A 5-year-old uncastrated male cat was taken to the HV-UESC small animal clinic, owing to a gradual increase in the volume of the nasal plane spanning over a year, accompanied by respiratory distress. Upon physical examination, the animal was found to have regular nutritional status, dehydration, hypocorous mucosae, and enlarged popliteal lymph nodes. In addition, multifocal areas of ulceration on the body as well as increased volume in the nasal plane (clown nose) were observed. Peripheral blood samples were collected for complete blood count and biochemical analysis. Due to clinical suspicion of fungal lesions, fine needle aspiration of the nasal lesion was performed and cytological slides were sent to the HV-UESC histopathology laboratory. Probable diagnoses included rhinosporidiosis, cryptococcosis, and histoplasmosis. The animal was subjected to thoracic radiography in the imaging division of the HV-UESC. Blood counts revealed thrombocytopenia (130,000/mm3) and biochemical tests presented hypoproteinemia (3.2 g/dL), with hypoalbuminemia (1.52 g/dL); however, chest radiography did not show pulmonary alterations. Results of cytological analysis indicated pyogranulomatous inflammation associated with intracellular organisms, demonstrating a pathogen morphology similar to that of Cryptococcus spp. Before the diagnosis, itraconazole 5 mg/kg (1 capsule, twice a day for 30 days) and fipronil (1 ampoule 0.5 mL, on the back) were prescribed. Follow-up after 40 days showed a slight improvement in cutaneous lesions and respiration, and no other abnormalities were observed. At the follow-up, the owner disclosed that medication could not be administered at the prescribed frequency because of the animal often ran away from home and stayed without prescription. Due to unsatisfactory response to the first treatment, the dosage of itraconazole1 was increased to 10 mg/kg (1 capsule, twice a day, for 30 days) with a new regimen and follow-up after 15 days. However, the owner did not make a follow-up visit. A year later, after several attempts to contact the owner, we were notified that the treatment had been discontinued and the cat still presents with lesions. Discussion: Although cryptococcosis is not endemic, the cat roamed freely on the streets and may have been infected by inhaling the microorganism spores present in the environment. The clinical signs exhibited by the animal were consistent with the findings associated with Cryptococcus spp. infection in cats, especially the "clown nose" lesion. The cat, besides being dehydrated at the time of care, had a regular nutritional status. Cytology, the only diagnostic technique used in this case through which it was possible to visually identify the fungus, was definitive for the diagnosis of cryptococcosis. Although the prescribed treatment is one of the most indicated in such cases, it was not effective, possibly due to inadequate administration. The length of infection, discontinuation of therapy, and absence of follow-ups for clinical reassessment certainly contributed to an unfavorable prognosis

Flow cytometry reticulocyte counting using acridine orange: validation of a new protocol

Submitted by Nuzia Santos ([email protected]) on 2015-03-26T13:19:34Z No. of bitstreams: 1 2014_170.pdf: 266255 bytes, checksum: 231d8aaac025e76936cafbd975f83c8c (MD5)Approved for entry into archive by Nuzia Santos ([email protected]) on 2015-03-26T13:19:49Z (GMT) No. of bitstreams: 1 2014_170.pdf: 266255 bytes, checksum: 231d8aaac025e76936cafbd975f83c8c (MD5)Approved for entry into archive by Nuzia Santos ([email protected]) on 2015-03-26T13:26:11Z (GMT) No. of bitstreams: 1 2014_170.pdf: 266255 bytes, checksum: 231d8aaac025e76936cafbd975f83c8c (MD5)Made available in DSpace on 2015-03-26T13:26:11Z (GMT). No. of bitstreams: 1 2014_170.pdf: 266255 bytes, checksum: 231d8aaac025e76936cafbd975f83c8c (MD5) Previous issue date: 2014Universidade Federal de Minas Gerais. Faculdade de Farmácia. Belo Horizonte, MG, Brasil/Prefeitura de Belo Horizonte. Belo Horizonte, MG, Brasil.Universidade Federal de Minas Gerais. Faculdade de Farmácia. Departamento de Análises Clinicas e Toxicologicas. Belo Horizonte, MG, BrasilUniversidade Federal de Minas Gerais. Faculdade de Farmácia. Departamento de Análises Clinicas e Toxicologicas. Belo Horizonte, MG, BrasilUniversidade Federal de Minas Gerais. Faculdade de Farmácia. Belo Horizonte, MG, BrasilCentro Universitário Newton Paiva. Belo Horizonte, MG, BrasilLaboratório de Patologia Clínica São Paulo. Serviço de Citometria. Belo Horizonte, MG, BrasilFundação Hemominas. Hematologia. Belo Horizonte, MG, BrasilLaboratório de Patologia Clínica São Paulo. Hematologia Clinica. Belo Horizonte, MG, BrasilFundação Oswaldo Cruz. Centro de Pesquisa Rene Rachou. Belo Horizonte, MG, BrasilFundação Oswaldo Cruz. Centro de Pesquisa Rene Rachou. Belo Horizonte, MG, BrasilFundação Oswaldo Cruz. Centro de Pesquisa Rene Rachou. Belo Horizonte, MG, BrasilIntrodução: Atualmente, a contagem de reticulócitos representa um desafio para os laboratórios clínicos no Brasil, principalmente os de pequeno e médio porte, nos quais ainda se utiliza o método manual. Este método apresenta algumas limitações, classificando-se como tedioso, demorado e de baixa precisão. Objetivos: O presente estudo desenvolveu e avaliou o desempenho de um novo protocolo laboratorial para contagem de reticulócitos por citometria de fluxo (CF) utilizando acridine orange (AO) como corante, visando padronizar um protocolo preciso, de fácil e rápida execução e custo acessível. Após a padronização do novo protocolo desenvolvido (CF/AO), fez-se a comparação com o método manual. Os resultados foram analisados de acordo com recomendações do National Committee for Clinical Laboratory Standards (NCCLS), atualmente Clinical and Laboratory Standards Institute (CLSI), para avaliar a intercambialidade entre os métodos, por meio da análise de regressão linear e teste t pareado, além de outros testes de controle de qualidade. Conclusão: Diante dos resultados obtidos referentes à correlação entre os métodos e os testes voltados ao controle de qualidade, pode-se admitir que o CF/AO estabelecido para contagem de reticulócitos possui vantagens inegáveis quando comparado com o método manual.Introduction: Currently, the reticulocyte counting is a challenge for clinical laboratories in Brazil, mainly for the ordinary ones, which still use the manual method. This method has some limitations, since it consists of a laborious method, time consuming, with low accuracy. Objectives: This study has developed and evaluated the performance of a New Laboratory Protocol for flow cytometry (FC) reticulocytes counting using acridine orange (AO) as dye, aiming to standardize a more precise, easy, fast implementation, and low cost protocol. After standardization of the New Protocol (FC/AO), it was compared with the manual method. The results were analyzed according to the recommendations of the National Committee for Clinical Laboratory Standards (NCCLS), now known as Clinical and Laboratory Standards Institute (CLSI), to evaluate the interchangeability of methods in linear regression analysis and paired t test, besides other quality control tests. Conclusion: Based on these results concerning to the correlation between the methods and the tests related to quality control, we can admit that FC/AO for reticulocyte counting shows undeniable advantages when compared to the preexisting manual method