Genotyping of news variants of the avian infectious bronchitis virus from Tolima department, Colombia / Genotipificación de variantes del virus de bronquitis infecciosa aviar en el departamento del Tolima, Colombia

ABSTRACT Objective. The aim of this study was identify the different genotypes of infectious bronchitis virus (IVB) present in commercial poultry farms from different localities of the Tolima Department, Colombia. Materials and methods. 105 samples of tracheal swabs of poultry of 21 farms were collected. Poultry had been vaccinated against IVB. An screen to identify positive samples and posteriorly the sequencing of the partial region of the S1 subunit and phylogenetic analysis of the isolates with the reference strains, including the vaccine currently used in the country was performed. Results . Poultry all farms had respiratory signs, but only four farms was confirmed the disease. Positive samples of the IBV (HT6, HT9, HT10 and HT11) were pathogenic for embryos 9-days-old. The HT6 sample was grouped in the same cluster that the Massachusetts strains. The HT9 and HT11 samples showed 99% similarity and were grouped genetically distant from the reference strains and other isolated. The HT10 sample showed low similarities with the isolates and reference strains, grouping alone in another cluster. Conclusions . New genotypes are circulating in the Tolima Department, where there is a risk of genetic recombination. Is believed that vaccines used were not providing cross-protection against the new genotypes.

RESUMEN Objetivo. El objetivo de este estudio fue identificar los diferentes genotipos del virus de bronquitis infecciosa (VBI) presente en granjas avícolas comerciales de diferentes localidades del departamento de Tolima, Colombia. Materiales y métodos . Se recolectaron 105 muestras de hisopados traqueales provenientes de aves de 21 granjas. Las aves habían sido vacunadas contra el VBI. Se realizó un "screen" para identificar muestras positivas y posteriormente la secuenciación de la región parcial de la subunidad S1 y el análisis filogenético de los aislamientos con las cepas de referencia, incluidas las vacunas utilizadas actualmente en el país. Resultados. Aves de todas las granjas tenían signos respiratorios, pero sólo cuatro granjas confirmaron la enfermedad. Las muestras positivas de VBI (HT6, HT9, HT10 y HT11) fueron patogénicas para embriones de 9 días de edad. La muestra HT6 se agrupó en el mismo clúster que las cepas de la vacuna Massachusetts. Las muestras HT9 y HT11 mostraron 99% de similitud y agrupan genéticamente distantes de las cepas de referencia y de los aislados. El HT10 mostró baja similitud con aislamientos y cepas de referencia, agrupándose separadamente en otro clúster. Conclusiones. Nuevos genotipos circulan en el departamento del Tolima, donde hay un riesgo de recombinación genética. Se estima que las vacunas utilizadas no están ofreciendo una protección cruzada contra los nuevos genotipos.

Analysis of the Bone Marrow Aspirates with Automated Hematology Analyzer for Assessment of the Bone Marrow Cellularity and Effective Hematopoiesis / 대한진단검사의학회지

BACKGROUND: Microscopic examination of the bone marrow (BM) smear has been a major method for the diagnostic and post-therapeutic evaluation of hematologic disease but is laborious and imprecise due to small number of cells counted. Recently, automated reticulocyte counting is available by the automated hematology analyzer. We analyzed the bone marrow aspirates using Coulter GEN S (GEN S) automated hematology analyzer and compared the results with those by the microscopic examination. METHODS: Total nucleated cells (TNC), leukocyte subpopulations, red cell count, hemoglobin and reticulocyte indices of the peripheral blood (PB) and the BM aspirates, were measured by GEN S in 392 samples including 142 normal control samples. Differential counts by microscopic examination of Wright stained BM films were used as a reference. RESULTS: TNC of the BM obtained by automated hematology analyzer correlated with the BM cel-lularity estimated by microscopic examination (r=0.587, P=0.000). The differential counts of neutrophils and monocytes correlated between these two methods (r=0.582, P=0.000, r=0.309, P=0.000). In acute leukemia, TNC of the PB and the BM, and the BM lymphocyte fraction were increased and the BM neutrophil fraction was decreased. In chronic myelogenous leukemia, TNC of the PB and the BM were high but distribution of leukocyte subpopulations was normal. In normal control group, the number of erythroid precusors correlated with the percentages of reticulocyte in the PB (r=0.425, P=0.000), and in patients with increased erythropoiesis, it showed strong correlation with immature reticulocyte fraction (IRF) of the PB (r=0.708, P=0.033). In aplastic anemia, IRF of the PB was reversely correlated to hemoglobin level, but in myelodysplastic syndrome, reticulocyte indices of the PB and the BM had no correlation with hemoglobin level. In patients with increased erythropoiesis, the percentages of reticulocyte in the PB were increased and those of the BM were decreased in proportion to reduction of hemoglobin level in the PB. CONCLUSIONS: Analysis of the BM aspirates using automated hematology analyzer will be useful in screening of pathological hematologic diseases and in estimating the bone marrow cellularity objectively before those by the microscopic examination. In anemia, this study could provide an additional information to evaluate the ineffective hematopoiesis using reticulocyte indices of the PB and the BM.

Performance Analysis of the Beckman Coulter GEN-S Automated Hematology Analyzer / 대한임상병리학회지

BACKGROUND: The complete blood cell count (CBC) and leukocyte differential counts are useful tools for making a diagnosis, treating and monitoring a disease. This study evaluated the perfor-mance of Beckman-Coulter's new model, the Coulter GEN-S system (Beckman Coulter Corpora-tion, Miami, USA; GEN-S), and compared it with the Sysmex NE-8000 (Sysmex Corporation, Kobe, Japan; NE-8000) and Sysmex R-3000 (Sysmex Corporation, Kobe, Japan; R-3000). METHODS: One hundred and four blood samples and 120 blood samples were randomly chosen for a comparison analysis from various in-patients and healthy persons, who visited Samsung med-ical center, respectively. The GEN-S system was evaluated according to the linearity and how well it compared with the NE-8000 in terms of the CBC, and the efficiency of the leukocyte suspect flags. RESULTS: The GEN-S showed that the determination coefficients (R(2)) of the WBC, RBC, platelet count and hemoglobin level were more than 0.94 (P 0.95 (P < 0.001) when compared with the NE-8000. In addition, the correlations of the leukocyte differential counts for neutrophils, eosinophils, and lymphocytes were good, but those for monocytes and basophils were poor. The sensitivity, specificity and positive predictive value of the leukocyte suspect flags of the GEN-S system were 89%, 73%, and 43%, respectively. CONCLUSIONS: These results demonstrate the comparable performance of the GEN-S system in clinical laboratories. However, a separate microscopic differential count is required due to the low positive predictive value for the leukocyte suspect flags.