Verificación del desempeño general del método Elecsys® Chagas en la detección de anticuerpos específicos contra Trypanosoma cruzi / Verification of the general performance of the Elecsys® Chagas method in the detection of specific antibodies against Trypanosoma cruzi

Resumen La enfermedad de Chagas es una parasitosis producida por Trypanosoma cruzi, prevalente principalmente en el continente americano, y observada en regiones no endémicas, producto de viajes y migraciones. El objetivo de este estudio fue comparar el desempeño del ensayo Elecsys® Chagas (Roche Diagnostics Alemania) (ECLIA) para el diagnóstico de la infección chagásica crónica con el método estándar y evaluar su posible empleo en reemplazo del método automatizado existente. Se estudiaron 77 muestras de sueros pertenecientes a pacientes con diagnóstico presuntivo de enfermedad de Chagas, procesadas por los distintos métodos disponibles en la Sección Parasitología del Hospital Muñiz: inmunoensayo quimioluminiscente de micropartículas (CMIA) (Abbott), enzimoinmunoanálisis de adsorción (ELISA) (Wiener) y hemaglutinación indirecta (HAI) (Lab. Lemos S.R.L.). Los resultados de los métodos ELISA y HAI fueron comparados con los obtenidos en la prueba ECLIA, y estos a su vez con el método automatizado disponible. De las muestras analizadas, 22 (28,57%) presentaron IgG anti-T. cruzi y 55 (71,43%) resultaron negativas. Con el método ECLIA se logró un 100% en los parámetros de desempeño, con diferencias en los intervalos de confianza. La razón de verosimilitud positiva y la razón de verosimilitud negativa clasificaron al ensayo como excelente y la potencia global del test apoyó esa afirmación. Los métodos inmunológicos automatizados ayudan a la performance diagnóstica en la etapa crónica de la enfermedad de Chagas, permiten minimizar errores, favorecen la velocidad de emisión de los resultados y, debido a su alta sensibilidad y especificidad, en ciertos escenarios podrían proponerse para usar como única técnica.

Abstract Chagas disease is a parasitosis caused by Trypanosoma cruzi, prevalent mainly in the American continent, and observed in non-endemic regions as a result of travel and migration. The objective of this study was to compare the performance of the Elecsys® Chagas (Roche Diagnostics Alemania) (ECLIA) assay for the diagnosis of chronic Chagas infection with the diagnostic standard, and to evaluate its possible use as a replacement for the existing automated method. A total of 77 serum samples belonging to patients with a presumptive diagnosis of Chagas disease were evaluated, processed by the different methods available in the Parasitology Section of Hospital Muñiz: microparticle chemiluminescent immunoassay (CMIA) (Abbott), enzyme-linked immunosorbent assay (ELISA) (Wiener) and indirect hemagglutination (HAI) (Lab. Lemos S.R.L). The results of the ELISA and HAI methods were compared with those obtained in the ECLIA test, and these in turn with the available automated method. Of the samples analysed, 22 (28.57%) presented IgG anti-T. cruzi and 55 (71.43%) were negative. With the ECLIA method, 100% was achieved in the performance parameters, with differences in the confidence intervals. The positive likelihood ratio and the negative likelihood ratio classify the essay as excellent, and the overall power of the test supports this statement. Automated immunological methods help diagnostic performance in the chronic stage of Chagas disease, allow minimising errors, favour the speed of issuance of results, and due to the high sensitivity and specificity, in certain scenarios, they could be proposed for use as single technique.

Resumo A doença de Chagas é uma parasitose causada pelo Trypanosoma cruzi, prevalente principalmente no continente americano, e observada em regiões não endêmicas em decorrência de viagens e migrações. O objetivo deste estudo foi comparar o desempenho do ensaio Elecsys® Chagas (Roche Diagnostics Alemanha) (ECLIA) para o diagnóstico da infecção crônica de Chagas com o método padrão e avaliar seu possível uso em substituição do método automatizado existente. Foram avaliadas 77 amostras de soro pertencentes a pacientes com diagnóstico presuntivo de doença de Chagas, processadas pelos diferentes métodos disponíveis na Seção de Parasitologia do Hospital Muñiz: imunoensaio quimioluminescente de micropartículas (CMIA) (Abbott), ensaio imunoenzimático de adsorção (ELISA) (Wiener) e hemaglutinação indireta (HAI) (Lab. Lemos S.R.L). Os resultados dos métodos ELISA e HAI foram comparados com os obtidos no teste ECLIA, e estes por sua vez com o método automatizado disponível. Das amostras analisadas, 22 (28,57%) apresentaram IgG anti-T. cruzi e 55 (71,43%) foram negativos. Com o método ECLIA, foram obtidos 100% nos parâmetros de desempenho, com diferenças nos intervalos de confiança. A razão de verossimilhança positiva e a razão de verossimilhança negativa classificam o ensaio como excelente, e a potencia geral do teste conformou essa afirmação. Os métodos imunológicos automatizados auxiliam no desempenho diagnóstico na fase crônica da doença de Chagas, permitem minimizar erros, favorecem a rapidez na emissão dos resultados e, devido à alta sensibilidade e especificidade, em determinados cenários, poderiam ser propostos para uso como técnica única.

Hemagglutination in gill capillaries of sheepshead, Archosargus probatocephalus (Perciformes: Sparidae), infected by a myxosporidean / Hemaglutinação em capilares branquiais de sargo-de-dente, Archosargus probatocephalus (Perciformes: Sparidae) infectado por um mixosporídeo

Abstract During a survey Myxozoa, four specimens of the sheepshead (18 ± 1.5 cm and 59 ± 2.5 g) (Archosargus probatocephalus) were collected in the Ipioquinha river (Maceió/AL). Transmission electron microscopy observations revealed erythrocyte agglutinations in gill capillaries located near spherical cysts containing myxospores of the genus Henneguya. This hemagglutination partially or totally obstructed the gill capillaries. Erythrocytes occurred in close adherence to each other, with a closed intercellular space. A few lysed erythrocytes were observed among agglutinated cells. The reduced lumen of the capillaries was partially filled with amorphous dense homogenous material adhering to the erythrocytes. In addition, heterogeneous masses of irregular lower electron density were observed in the reduced channel of the capillary. The agglutinated erythrocytes appeared dense and homogenous, lacking cytoplasmic organelles. The nuclei had the appearance of normal condensed chromatin masses, generally without visible nucleoli. This occurrence of hemagglutination only in the capillaries located in close proximity to the developing myxozoan cysts suggests that parasite development may be a factor triggering erythrocyte agglutination. This is supported by previous experimental studies that showed a probable correlation between parasitic infections and hemagglutination. Nonetheless, further studies are necessary in order to better understand the physicochemical processes involved in this phenomenon.

Resumo Durante pesquisa de mixozoários foram coletados quatro espécimes do peixes sargo-de-dente (18 ± 1.5 cm e 59 ± 2.5 g) (Archosargus probatocephalus), no rio Ipioquinha (Maceió/AL). Observações por microscopia eletrônica de transmissão revelaram aglutinação de eritrócitos em capilares branquiais localizados próximos a cistos esféricos, contendo mixosporos do gênero Henneguya. Essa hemaglutinação obstruiu parcial ou totalmente os capilares branquiais. Os eritrócitos apareceram em forte aderência entre si, com espaço intercelular fechado. Foram observados poucos eritrócitos lisados entre as células aglutinadas. O lúmen reduzido dos capilares foi parcialmente preenchido com material homogêneo denso amorfo aderido aos eritrócitos, além de massas livres heterogêneas de densidade eletrônica baixa e irregular observadas no canal reduzido dos capilares. Os eritrócitos aglutinados pareciam densos e homogêneos, sem organelas citoplasmáticas. Os núcleos apareceram como massas normais de cromatina condensada, geralmente sem nucléolos visíveis. A ocorrência de hemaglutinação apenas nos capilares, localizados nas proximidades dos cistos mixozoários, sugere que o desenvolvimento parasitário pode ser um fator desencadeante da aglutinação eritrocitária. Isso é corroborado por estudos experimentais anteriores que mostraram uma provável correlação entre infecções parasitárias e hemaglutinação. No entanto, novos estudos são necessários para melhor compreender os processos físico-químicos envolvidos neste fenômeno.

Diagnostic efficacy of indirect haemagglutination assay for detection of Schistosoma japonicum infections among boatmen and fishermen in the Dongting Lake region / 中国血吸虫病防治杂志

OBJECTIVE@#To evaluate the diagnostic efficacy of indirect haemagglutination assay (IHA) for detection of Schistosoma japonicum infections among boatmen and fishermen in Dongting Lake region, so as to provide insights into improving the schistosomiasis surveillance program among boatmen and fishermen.@*METHODS@#The boatmen and fishermen were detected for S. japonicum infections using IHA and Kato-Katz technique or miracidium hatching test nylon gauze simultaneously at schistosomiasis testing sites in the anchor sites for boatmen and fishermen in the Dongting Lake region during the period from 2014 to 2016, and using IHA for serological screening followed by parasitological testing of seropositives during the period from 2017 to 2019. The sensitivity and specificity of IHA were evaluated for detection of S. japonicum infections among boatmen and fishermen, with the 2014-2016 parasitological testing results as a gold standard. In addition, the seroprevalence of S. japonicum infections was compared among boatmen and fishermen with different characteristics and among years.@*RESULTS@#A total of 306 schistosomiasis testing sites were assigned for boatmen and fishermen, and a total of 143 360 person-time boatmen and fishermen were tested for S. japonicum infections in the Dongting Lake region from 2014 to 2019. The sensitivity and specificity of IHA were 69.9%, 97.3% and 96.1% (χ2 = 74.6, P < 0.05), and 70.9%, 74.5% and 71.9% for detection of S. japonicum infections from 2014 to 2016 (χ2 = 29.4, P < 0.05), respectively. The seroprevalence of S. japonicum infections reduced from 30.3% in 2014 to 1.8% in 2019 among boatmen and fishermen, appearing an overall tendency towards a decline (Z = 1 552.4, P < 0.05). In addition, male, individuals at ages of 45 to 60 years, full-time boatmen and fishermen were more likely to be seropositive for S. japonicum infections (all P values < 0.05).@*CONCLUSIONS@#The seroprevalence of S. japonicum infections appeared a tendency towards a decline among boatmen and fishermen in the Dongting Lake region year by year from 2014 to 2019. IHA presented a high efficacy for screening of S. japonicum infections among boatmen and fishermen in the Dongting Lake region.

Serologic strategy in detecting RHD altered alleles in Brazilian blood donors

ABSTRACT Background: We evaluated different technological approaches and anti-D clones to propose the most appropriate serologic strategy in detecting the largest numbers of D variants in blood donors. Methods: We selected 101 samples from Brazilian blood donors with different expressions of D in our donor routine. The tests were performed in immediate spin (IS) with eleven commercially available anti-D reagents in a tube and microplate. The D confirmatory tests for the presence of weak D included the indirect antiglobulin test (IAT) in a tube, gel and solid-phase red blood cell adherence (SPRCA). All DNA samples were extracted from peripheral blood and the D variants were classified using different molecular assays. Results: The RHD variants identified by molecular analysis included weak D types (1, 2, 3, 11 and 38) and partial Ds (DAR1.2, DAR1, DAR3.1, DAU0, DAU2, DAU4, DAU5, DAU6, DMH and DVII). The monoclonal-monoclonal blend RUM-1/MS26 was the best anti-D reagent used in detecting the D antigen in the IS phase in a tube, reacting with 83.2% of the D variants, while the anti-D blend D175 + 415 was the best monoclonal antibody (MoAb) used in a microplate to minimize the need for an IAT, reacting with 83.2% of the D variants. The D confirmatory tests using SPRCA showed a reactivity (3 - 4+) with 100% of the D variant samples tested. Conclusion: Our results show that, even using sensitive methods and MoAbs to ensure the accurate assignment of the D antigen, at least 17% of our donor samples need a confirmatory D test in order to avoid alloimmunization in D-negative patients.

Effect of nabumetone on humoral immune responses in mice / Efeito da nabumetona nas respostas imunes humorais em camundongos

Nabumetone is used to reduce the pain and inflammation in rheumatoid arthritis. In the current study, immunomodulatory effect of Nabumetone is investigated in mice. The control group was administered normal saline orally as placebo. Nabumetone was administered orally via gavage in two treatment groups at 14mg/kg.b.w. doses and 28mg/kgb.w., respectively. Haemagglutination (HA) assay, Jerne hemolytic plaque and mice lethality assays were applied. In HA assay, the titer was significantly decreased in Nabumetone treatment groups (P< 0.001). In Jerne hemolytic plaque formation assay, there was a significant reduction (P< 0.001) in number of plaques in Nabumetone treated groups when compared with control. In mice lethality assay, there was a significant difference in mortality ratio of mice in control and Nabumetone treated groups (P< 0.001). Therefore, it is concluded that Nabumetone suppresses the humoral immune response in mice.(AU)

A nabumetona é usada na redução da dor e inflamação da artrite reumática. No presente estudo, o efeito imunomodulador é investigado em camundongos. O grupo de controle recebeu solução salina via oral como placebo. Nabumetona foi administrada oralmente via gavagem em dois grupos de tratamentos com doses de 14mg/kg.b.w. e 28mg/kgb.w., respectivamente. Foram realizados ensaios de hemaglutinação (HA), placa hemolítica de Jerne e letalidade dos camundongos. No ensaio HA, o grau foi significativamente menor nos grupos de tratamento com nabumetoma (P< 0.001). No ensaio de formação de placa hemolítica de Jerne houve redução significativa (P< 0.001) no número de placas em grupos tratados com nabumetoma comparado ao controle. No ensaio de letalidade dos camundongos houve diferença significativa no grau de mortalidade de camundongos no grupo de controle e grupos tratados com nabumetoma (P< 0.001). Portanto, conclui-se que a Nabumetoma suprime a resposta imune humoral em camundongos.(AU)

Esophageal manometry in systemic sclerosis: findings and association with clinical manifestations

SUMMARY INTRODUCTION Systemic sclerosis (SSC) is an autoimmune disorder that affects several organs of unknown etiology, characterized by vascular damage and fibrosis of the skin and organs. Among the organs involved are the esophagus and the lung. OBJECTIVES To relate the profile of changes in esophageal electromanometry (EM), the profile of skin involvement, interstitial pneumopathy (ILD), and esophageal symptoms in SSC patients. METHODS This is an observational, cross-sectional study carried out at the SSC outpatient clinic of the Hospital de Clínicas of the Federal University of Uberlândia. After approval by the Ethics Committee and signed the terms of consent, 50 patients were initially enrolled, from 04/12/2014 to 06/25/2015. They were submitted to the usual investigations according to the clinical picture. The statistical analysis was descriptive in percentage, means, and standard deviation. The Chi-square test was used to evaluate the relationship between EM, high-resolution tomography, and esophageal symptoms. RESULTS 91.9% of the patients had some manometric alterations. 37.8% had involvement of the esophageal body and lower esophageal sphincter. 37.8% had ILD. 24.3% presented the diffuse form of SSC. No association was found between manometric changes and clinical manifestations (cutaneous, pulmonary, and gastrointestinal symptoms). CONCLUSION The present study confirms that esophageal motility alterations detected by EM are frequent in SSC patients, but may not be related to cutaneous extension involvement, the presence of ILD, or the gastrointestinal complaints of patients.

RESUMO INTRODUÇÃO A esclerose sistêmica (ES) é uma doença autoimune que afeta vários órgãos de etiologia desconhecida, caracterizada por dano vascular e fibrose da pele e órgãos. Entre os órgãos envolvidos estão o esôfago e o pulmão. OBJETIVOS Relacionar o perfil das alterações na eletromanometria (ME), o perfil de acometimento da pele, a pneumopatia intersticial (PI) e os sintomas esofágicos em pacientes com ES. MÉTODO Trata-se de um estudo observacional, transversal, realizado no ambulatório de SSC do Hospital das Clínicas da Universidade Federal de Uberlândia. Após aprovação pelo Comitê de Ética e assinatura dos termos de consentimento, 50 pacientes foram inicialmente convidados, de 04/12/2014 a 25/06/2015. Eles foram submetidos às investigações usuais de acordo com o quadro clínico. A análise estatística foi descritiva em porcentagem, média e desvio padrão. O teste Qui-quadrado foi utilizado para avaliar a relação entre ME, tomografia de alta resolução e sintomas esofágicos. RESULTADOS 91,9% dos pacientes apresentaram alterações manométricas. 37,8% tinham envolvimento do corpo esofágico e do esfíncter esofágico inferior. 37,8% tinham IP. 24,3% apresentaram a forma difusa da ES. Não há associação entre alterações manométricas e manifestações clínicas (sintomas cutâneos, pulmonares e gastrointestinais). CONCLUSÃO O presente estudo confirma que as alterações da motilidade esofágica detectadas pela EM são frequentes em pacientes com SSC, mas podem não estar relacionadas ao envolvimento cutâneo, à de DPI ou às queixas gastrointestinais dos pacientes.

Valeurs de référence de l'hémogramme chez les donneurs de sang du Centre National de Transfusion Sanguine de Libreville

Introduction : les valeurs de référence peuvent être obtenues à partir des bases de données. Nous nous proposons de produire ces valeurs à partir d'une base de données hématologiques et de souligner les contraintes inhérentes à cette méthodologie. Matériel et méthodes: Il s'agit d'une étude rétrospective, réalisée du 1er janvier 2015 au 30 mars 2017, au Centre National de Transfusion Sanguine de Libreville. Les dossiers des donneurs, dont les sérologies (HIV, AgHBs, Ac HVC, VDRL et TPHA) étaient négatives ont été retenus. Les paramètres de l'hémogramme ont été notés pour chaque dossier. L'identification des valeurs aberrantes a été réalisée à l'aide de la méthode de Cook, la nature de la distribution a été étudiée grâce au test de normalité de Shapiro-Wilk. Les limites de référence à 95% et leurs intervalles de confiance à 90% ont été calculés. La recherche des partitions a été effectuée à l'aide du z-test. Les différences étaient considérées comme statistiquement significatives, pour une valeur de p inférieure à 0,05. Résultats: ce protocole a concerné 27022 dossiers, dont 2013 (7,4%) ont été exploités. L'intervalle de référence à 95% des leucocytes allait de 2,8.103 à 6,1.103/mm3, alors que celui des hématies allait de 4,3.106 à 5,3.106/mm3et celui des plaquettes de 129,7.103 à 258,0.103/mm3. De plus, les hématies allaient de 4,4.106 à 5,3.106/mm3chez les hommes contre 4,4.106 à 5,1.106/mm3pour les femmes de même tranche d'âge.Conclusion: la réalisation d'une telle étude nécessite une collaboration interdisciplinaire, une bonne tenue des bases de données et nous rappelle la primauté d'une différence biologique, par rapport à une différence statistique

Assessment of hemagglutination activity of porcine deltacoronavirus

Comparison of antigenic mutation during egg and cell passage cultivation of H3N2 influenza virus

Blood typing in positive DEA 1 dogs: comparative analysis between immunochromatography, hemagglutination and flow cytometry / Tipificação sanguínea em cães AEC 1 positivo: análise comparativa entre a imunocromatografia, hemoaglutinação e citometria de fluxo

Blood typing techniques have been improved to ensure greater safety for transfusion procedures. Typification for the DEA 1 antigen through flow cytometry should offer more reliability to routine immunohematology in donor and recipient dogs. Currently, the DEA 1 group is starting to be an autosomal dominant allelic system with the DEA 1 negative type and its variations of positivity. The present study investigated the DEA 1 antigen using the techniques of immunochromatography, hemagglutination and flow cytometry. Among the positive animals for the DEA 1 group, typified by flow cytometry, medium intensities of fluorescence were found, which are indicative of weak, moderate and strong antigenicity. This enabled the division of the DEA 1 group into weak positive, moderate positive and strong positive. The blood typing techniques for the DEA 1 group by flow cytometry, agglutination and immunochromatography had positive (Spearman r=0.70) and statistically significant (p>0.0001) correlations.(AU)

As técnicas de tipificação sanguínea vêm sendo aperfeiçoadas para garantir maior segurança aos procedimentos transfusionais. A tipificação para o antígeno AEC 1 com o emprego da citometria de fluxo poderá oferecer mais confiabilidade à rotina da imunohematologia em cães doadores e receptores. Na atualidade, o grupo AEC 1 passou a ser denominado como um sistema alélico autossômico dominante com o tipo AEC 1 negativo e suas variações de positividade. O presente trabalho comparou os resultados de três técnicas utilizadas para a pesquisa do antígeno AEC 1: cromatografia; hemoaglutinação e citometria de fluxo. Dentro dos indivíduos positivos para o grupo AEC 1, tipificados pela citometria de fluxo, foram encontradas intensidades médias de fluorescência indicadoras de antigenicidade fraca, moderada e forte, podendo-se dividir o grupo AEC 1 em positivo fraco, positivo moderado e positivo forte. As técnicas de tipificação sanguínea para o grupo AEC 1 por cromatografia, hemoaglutinação e citometria de fluxo apresentaram correlação positiva (Spearman r=0,70) e estatisticamente significativa (p<0,0001).(AU)

Avaliação das reações de Hemaglutinação Indireta (HI) e Aglutinação Modificada (MAT) na detecção de anticorpos IgG anti-T. gondii em exsudatos cárneos bovinos / Evaluation of indirect Hemagglutination (HI) and Modified Agglutination (MAT) reactions in the detection of anti-T. gondiiIgG antibodies in bovine meat exudates

Toxoplasmose é uma zoonose parasitária com ampla distribuição mundial provocada pelo Toxoplasma gondii, considerado um dos protozoários mais bem sucedidos do planeta, pois infecta cerca de um terço da população mundial. Dentre as formas de transmissão, o consumo de carne mal cozida, contendo cistos, tem sido considerado um fator de risco para aquisição desta zoonose. Uma abordagem alternativa para o controle da toxoplasmose pela ingestão de carne bovina seria a sorologia dos bovinos, já que animais soropositivos albergam cistos teciduais. Contudo, a obtenção de soro para esta avaliação, nem sempre é factível, dada a dificuldade de coleta de sangue durante a linha de abate e sua ausência em cortes comerciais. O exsudato cárneo é uma alternativa para detecção de anticorpos anti - T. gondii em cortes comerciais de carne, que foi a proposta deste estudo para avaliar o desempenho dos testes de Hemaglutinação Indireta (HI) e Aglutinação Modificada (MAT) quando comparados ao ELISA usando exsudato cárneo. Este estudo mostrou que a acurácia dos testes de aglutinação não foi viável devido aos baixos índices de sensibilidade e especificidade quando comparados ao ELISA. Estes dados demonstram a importância da escolha de testes eficientes como ELISA para aplicação no controle da qualidade e inocuidade de cortes comerciais de carne bovina. (AU)

Toxoplasmosis is a parasitic zoonosis with a wide worldwide distribution caused by Toxoplasmagondii, which is considered one of the most successful protozoa on the planet, since it can infect a third of the world population. Among the forms of transmission, consumption of undercooked meat has been considered as a risk factor for the acquisition of this zoonosis. An alternative approach to toxoplasmosis control by beef ingestion could be the serological diagnosis in cattle, since seropositives animals harbor tissue cysts. However, the use of serum for this evaluation is not always feasible due to the difficulty of blood collection during slaughter and its absence in commercial beef cuts. Meat exudate is an alternative for the detection of anti-T. gondii antibodies in commercial beef cuts, which was the propose of this study to evaluate the performance of Indirect Hemagglutination (HI) and Agglutination Modified (MAT) tests compared to ELISA using meat exudates. This study showed that the agglutination tests accuracy was not viable due to low sensitivity and specificity indexes when compared to ELISA. These data demonstrate the importance of choosing accurate tests such as ELISA for application in quality control and safety of commercial beef cuts. (AU)

In vivo and in vitro Pharmacological Evaluation of Gymnosporia Royleana / Evaluación Farmacológica de Gymnosporia Royleana in vivo e in vitro

ABSTRACT Objective: Gymnosporia royleana (G royleana) Wall ex MA Lawson, locally known as (Sur Azghee), is traditionally used for the management of various diseases. In the current investigation, we made an effort to scientifically validate its traditional use in various pathological conditions, such as microbial infections and cancer, and to explore its additional pharmacological activities via random screening against locally accessible pharmacological methods, irrespective of its traditional uses like antidiabetic, haemagglutination and antioxidant assays. Methods: Extraction was carried out using a cold maceration methodology. Dilution method was used for antimicrobial susceptibility testing using different concentrations. Streptozocin (STZ) induced protocol was used to assess antidiabetic activity at a dose level of 200, 400 mg/ kg. Antioxidant activity, haemagglutination activity, and anticancer activities against HepG-2 and MCF-7 cell lines were determined as per established protocols. Similarly, the maximum amount of phenolic content (12.02 mg 100 g) was determined by using Folin Ciocalteu assay. Results: Promising antimicrobial activities in terms of minimum inhibitory concentration (MIC) were noted for crude extract (25-200 µg/mL), n-hexane (100-400 µg/mL), ethyl acetate (50-200 µg/mL) and aqueous (100-400 µg/mL). Antidiabetic potential was significant at a dose level of 200-400 mg/kg bodyweight by reducing the blood glucose level at days 10 and 15. The percentage of 2,2-diphenyl-1-picrylhydrazyl (DPPH) values increase by increasing the concentration of the plant extract (10-100 µg/mL). The methanol extract was found to possess high agglutination activity. Conclusion: It was concluded that this plant species possess significant antimicrobial, antidiabetic, antioxidant, anticancer and haemagglutination activities, which could be attributed to the phenolic content of the extract.

RESUMEN Objetivo: Gymnosporia royleana (G royleana) Wall ex MA Lawson, localmente conocida como "Sur Azghee", se utiliza tradicionalmente para el tratamiento de diversas enfermedades. En la investigación presente, tratamos de validar científicamente su uso tradicional en varias condiciones, tales como las infecciones microbianas y el cáncer, así como explorar sus actividades farmacológicas adicionales mediante el tamizado aleatorio frente a los métodos farmacológicos localmente accesibles, independientemente de sus usos tradicionales como ensayos antidiabéticos, hemaglutinantes, y antioxidantes. Métodos: La extracción se realizó mediante una metodología de maceración en frío. Un método de dilución se utilizó para la prueba de susceptibilidad antimicrobiana utilizando diferentes concentraciones. Se utilizó el protocolo inducido por estreptozotocina (STZ) para evaluar la actividad antidiabética a un nivel de dosis de 200, 400 mg/kg. La actividad antioxidante, la actividad de hemaglutinación, y las actividades anticancerígenas contra las líneas celulares HepG-2 y MCF-7, se determinaron según los protocolos establecidos. De modo similar. la cantidad máxima de contenido fenólico (12.02 mg 100 g) se determinó mediante el uso del ensayo Folin-Ciocalteu. Resultados: Se observaron actividades antimicrobianas prometedoras en términos de la concentración inhibitoria mínima (CIM) para el extracto crudo (25-200 μg/mL), el n-hexano (100-400 μg/mL), el acetato de etilo (50-200 μg/mL), y el extracto acuoso (100-400 μg/mL). El potencial antidiabético fue significativo a un nivel de dosis de 200-400 mg/kg de peso corporal mediante la reducción del nivel de glucosa en sangre a los 10 y 15 días. El porcentaje de los valores de 2,2-difenil-1-picrilhidracilo (DPPH) se incrementa al aumentar la concentración del extracto de la planta (10-100 μg/mL). Se halló que el extracto de metanol posee una alta actividad de aglutinación. Conclusión: Se concluyó que esta especie de planta posee importantes actividades antimicrobianas, antidiabéticas, antioxidantes, anticancerígenas y hemaglutinantes, que podrían atribuirse al contenido fenólico del extracto.

Development of a HA1-specific enzyme-linked immunosorbent assay against pandemic influenza virus A H1N1

PURPOSE: Enzyme-linked immunosorbent assay (ELISA) has been used in the diverse field to evaluate influenza virus infection; for the surveillance, diagnosis, efficacy evaluation, and development of the vaccine. The aim of this study was to establish an ELISA for detecting HA strain-specific antibodies using recombinant pandemic A H1N1 (pH1N1) HA1 (rHA1) protein. MATERIALS AND METHODS: rHA1 was produced in baculovirus system. The clinical performance of the developed ELISA was validated using human serum samples, by comparison with standard methods for detecting a neutralizing antibody; hemagglutination inhibition (HI) assay and microneutralization test (MNT). The ability of the ELISA system to evaluate the efficacy test of an influenza vaccine was explored by measuring antibody levels in the serum of vaccinated mice. RESULTS: Our ELISA could detect anti-rHA1 antibody in influenza-infected patients and vaccinated subjects. Compared to HI assay and MNT as reference methods, our method showed good performance in detection of anti-rHA1 antibody. Detection of the anti-rHA1 antibody in vaccinated mice and its correlation with titers in HI assay was also proved in a mice model. CONCLUSION: An ELISA system using rHA1 of pH1N1 influenza virus was developed, and showed good clinical performance in diagnosis of influenza virus infection and evaluation of the vaccination efficacy in both human and animal models.

Antibody responses after vaccination against equine influenza in Korea in 2016–2018 / 대한수의학회지

Equine influenza (EI) is the main cause of respiratory illness in equines across the globe and is caused by equine influenza A virus (EIV-A), which has impacted the equine industry internationally because of the marginal mortality and high morbidity. In the present study, the immune responses after equine influenza vaccination were evaluated in 4,144 horses in Korea using the hemagglutination inhibition (HI) assay. The equine influenza virus (EIV), A/equine/South Africa/4/03 (H3N8), was used as the antigen in the HI assay. The mean seropositive rates were 89.2% (97.4% in 2016, 77.6% in 2017, and 92.4% in 2018). This paper highlights the advances in understanding the effects of vaccines and control strategies for mitigating the emerging menace by EIV.

Evaluation of Waning Immunity at 6 Months after Both Trivalent and Quadrivalent Influenza Vaccination in Korean Children Aged 6–35 Months

BACKGROUND: The titer of influenza vaccine-induced antibodies declines over time, and younger children have lower immunogenicity and shorter duration of immunity. This study aimed to compare persistence of antibody at 6 months after influenza vaccination according to influenza virus strains, vaccine type, antigen dose, and primed status in children aged 6 to 35 months. METHODS: A total 124 healthy children aged 6 to 35 months were enrolled from September to December 2016 at 10 hospitals in Korea and randomly assigned to either a full dose of quadrivalent influenza vaccine or a half dose of trivalent influenza vaccine with Victoria B strain group. Hemagglutination inhibition antibody titers (that measure the seroprotection rates) were assessed for the recommended influenza strains at 6 months post vaccination. RESULTS: The seroprotection rates at 6 months for strains A (H1N1), A (H3N2), B/Yamagata, and B/Victoria were 88.7%, 97.4%, 36.6%, and 27.6%, respectively. The seroprotection rates for A (H1N1), A (H3N2) and B (Victoria) were 91.4%, 98.7% and 27.5% in a full dose of quadrivalent vaccine vs. 83.7%, 94.6% and 27.9% in a half dose trivalent vaccine, respectively. The seroprotection rate for the B (Yamagata) strain was 23.8% in the quadrivalent group and 14.0% in the trivalent group. CONCLUSION: Persistence of antibodies at 6 months was more favorable against the influenza A strains than against the B strains. Persistence of antibodies to additional B strain at 6 months was superior in the quadrivalent vaccine group. The immunity of primed children with different B strains was not superior to that of the unprimed group with another B strain.

Recharacterization of the Canine Adenovirus Type 1 Vaccine Strain based on the Biological and Molecular Properties

Canine adenovirus type 1 (CAV-1) infection results in hepatitis in dogs. In this study, we investigated the biologic and genetic characteristics of the CAV-1 vaccine strain (CAV1V) to improve quality control about CAV vaccine. The identity of CAV1V as CAV-1 was confirmed based on its cytopathic effects and the results of hemagglutination (HA) and immunofluorescence assays, and electron microscopy. The CAV1V strain reached 10(7.5) TCID(50)/mL in MDCK cells at 4 days post-inoculation and exhibited hemmagglutination activity of 256 U using guinea pig erythrocytes. Intranuclear fluorescence in the infected cells was observed and typical adenoviruses were observed in electon microscope. CAV1V strain was identified as a CAV-1 strain by nucleotide sequence analysis. In a comparison of the nucleotide sequences of the fiber genes of several CAV strains, CAV1V showed the highest similarity (99.8%) with the GLAXO strain, which was isolated in Canada. Our biological characterization of CAV1V will facilitate quality control of the canine hepatitis vaccine.

Recombinant-attenuated Salmonella Pullorum strain expressing the hemagglutinin-neuraminidase protein of Newcastle disease virus (NDV) protects chickens against NDV and Salmonella Pullorum challenge

Newcastle disease virus (NDV) and Salmonella Pullorum have significant damaging effects on the poultry industry, but no previous vaccine can protect poultry effectively. In this study, a recombinant-attenuated S. Pullorum strain secreting the NDV hemagglutinin-neuraminidase (HN) protein, C79-13ΔcrpΔasd (pYA-HN), was constructed by using the suicide plasmid pREasd-mediated bacteria homologous recombination method to form a new bivalent vaccine candidate against Newcastle disease (ND) and S. Pullorum disease (PD). The effect of this vaccine candidate was compared with those of the NDV LaSota and C79-13ΔcrpΔasd (pYA) strains. The serum hemagglutination inhibition antibody titers, serum immunoglobulin G (IgG) antibodies, secretory IgA, and stimulation index in lymphocyte proliferation were increased significantly more (p 0.05). Moreover, the novel strain provides 60% and 80% protective efficacy against the NDV virulent strain F48E9 and the S. Pullorum virulent strain C79-13. In summary, in this study, a recombinant-attenuated S. Pullorum strain secreting NDV HN protein was constructed. The generation of the S. Pullorum C79-13ΔcrpΔasd (pYA-HN) strain provides a foundation for the development of an effective living-vector double vaccine against ND and PD.

Safety and Immunogenicity of an Egg-Cultivated Quadrivalent Inactivated Split-virion Influenza Vaccine (GC3110A) in Healthy Korean Children: a Randomized, Double-blinded, Active-controlled Phase III Study

BACKGROUND: The frequency with which the 2 B lineages have been found to cocirculate in a season has been on the rise, which has spurred the need for a quadrivalent influenza vaccine (QIV) to protect against both B lineages. The World Health Organization (WHO) recommended that QIV include both B lineages beginning in the 2013–2014 flu season. This study was conducted to evaluate the immunogenicity and safety of an egg-cultivated QIV in healthy Korean children and adolescents aged ≥ 6 months to < 19 years. METHODS: A total of 528 subjects were randomized 4:1 to receive either a QIV (GC3110A) or a trivalent influenza vaccine. Hemagglutination inhibition antibody responses were assessed 28 days after the last dose. Safety was also evaluated. RESULTS: The proportion of subjects in the GC3110A group who achieved seroconversion was confirmed to exceed 40% across all age groups. The proportion of subjects aged ≥ 6 months to < 3 years in the GC3110A group who achieved seroprotection failed to meet the Ministry of Food and Drug Safety (MFDS) standard of 70%. Potential causes may include the small number of subjects, as well as the small dosage. However, results pertaining to the other age groups satisfied the MFDS standard. The safety profile was also comparable to that of the control. CONCLUSION: The new quadrivalent split influenza vaccine may offer broader protection to children and adolescents aged ≥ 3 years to < 19 years of age against both influenza B lineages than the existing trivalent influenza vaccines (Registered at the ClinicalTrials.gov NCT02541253).

Isolation and identification of canine adenovirus type 2 from a naturally infected dog in Korea / 대한수의학회지

Canine adenovirus type 2 (CAV-2) infection results in significant respiratory illness in dogs. Isolating and culturing CAV-2 allows for investigations into its pathogenesis and the development of vaccines and diagnostic assays. In this study, we successfully isolated a virus from a naturally infected dog in Gyeonggi-do, Korea. The virus was propagated in Madin-Darby canine kidney (MDCK) and Vero cells and showed a specific cytopathic morphology that appeared similar to a bunch of grapes. The virus was first confirmed as CAV-2 based on these cytopathic effects, an immunofluorescence assay, hemagglutination assay, and electron microscopy. The viral titer of the isolate designated APQA1601 reached 10(6.5) 50% tissue culture infections dose per mL in MDCK cells and exhibited no hemagglutination units with erythrocytes from guinea pig. The virus was also confirmed by polymerase chain reaction and next-generation sequencing. The APQA1601 strain had the highest similarity (~99.9%) with the Toronto A26/61 strain, which was isolated in Canada in 1976 when the nucleotide sequences of the full genome of the APQA1601 strain were compared with those of other CAV strains. Isolating CAV-2 will help elucidate the biological properties of CAV-2 circulating in Korean dogs.

Serologic monitoring of animal welfare-oriented laying-hen farms in South Korea / 대한수의학회지

As animal welfare issue becomes important, the European Union bans conventional cages for laying hens from 2012. So the alternative housing systems like floor pens, aviaries or free range systems have been suggested. From 2011 to 2014, we monitored 20 welfare-oriented laying hen farms in South Korea to figure out serological status of major viral diseases. During this period, total 3,219 blood samples were collected from the randomly selected chickens to test and evaluate the hemagglutination inhibition titers for low pathogenic avian influenza, Newcastle disease and egg drop syndrome '76. A total of 2,926 blood samples were tested through enzyme linked immunosorbent assay (ELISA) to assess the serological status of infectious bronchitis (IB). The distribution of ELISA titers for IB was various from almost 0 to 20,000 through the all weeks of age. Also, the antibody coefficient of variation for most of the diseases in this study was higher than those of typical cage layers. As this study was the first surveillance for major avian viral diseases of the animal welfare-oriented farms in South Korea, the results obtained from this study will help to determine what information and resources are needed to maintain better biosecurity and to improve the health and welfare of laying hen flocks.