Portable near-infrared spectroscopy: A rapid and accurate blood test for diagnosis of Haemonchus contortus infection and for targeted selective treatment of sheep

Haemonchus contortus is the most prevalent and important gastrointestinal nematode (GIN) in small ruminants. Since it reduces the packed cell volume (PCV), causing anemia, early diagnosis can be used for targeted selective treatment (TST) of sheep, reducing antiparasitic drug use and anthelmintic resistance. This study aimed to predict PCV values through near-infrared spectroscopy (NIRS) and to develop a classification and diagnosis model of H. contortus infection using PCV values, eggs per gram of feces (EPG) counts and mean daily weight gain (DWG). A total of 1728 spectra were collected from blood samples of 216 lambs with a portable NIR spectroscope. In parallel, other parameters indicative of infection were measured: PCV by hematocrit, FAffa MAlan CHArt (FAMACHA) scores, EPG and DWG. To evaluate the relationship between NIRS spectra and the evaluated parameters, principal component analysis (PCA) was used for an exploratory analysis, regression by the partial least squares method (PLS) for the prediction of PCV values via NIRS, and PCA linear discriminant analysis (PCA-LDA) as a classification model for diagnosis. The absorption peaks in the NIRS region associated with the excitation of overtones of nitrogen-hydrogen (N-H) functional groups of proteins had a strong impact on the principal components (PCs), indicating that blood proteins, especially hemoglobin, can be estimated by the NIRS technique. The model for predicting PCV by PLS presented a standard error of prediction of 2.53, root-mean-square error of 2.48, and coefficient of determination of 0.84, indicating good correlation between the PCV values predicted by the model and the PCV obtained by hematocrit. The PCA-LDA model presented 93.33% sensitivity and 82.18% accuracy, both higher than those of the FAMACHA method, as was expected for resilient Morada Nova lambs. The multivariate models associated with the NIRS technique reported here can be used in the future as a quick and versatile tool for H. contortus infection diagnosis and TST application in lambs.     

An improved culture system for virus isolation and detection

Cell culture plays an important role in virology. It provides a platform for the detection and isolation of viruses as well as for the biochemistry and molecular biology based studies of viruses. In the present work, a new system that could permits multiple (different) cell lines to be simultaneously cultured in one dish was developed. In the system, each cell line was cultured in an isolated zone in the same dish or well and the system is therefore called an isolated co-culture system. The usefulness of this novel approach for virus isolation was demonstrated using a model system based on adenovirus.     

Analysis of the value of copper erythrocyte concentration measurement in the diagnosis of copper deficiency in bovines

BackgroundA reliable and practical method for assessing Cu status in live animals is not available. Blood Cu levels may not accurately reflect the true Cu status of the herd, and can over-predict Cu status during stress and inflammation. On the other hand, assessment of liver Cu is the most reliable indicator of Cu stores, but it is an invasive procedure that requires specialized training. The aim of this study was to evaluate the usefulness of Cu levels in red blood cells to determine the Cu status, with special emphasis in their correlation with erythrocyte Cu, Zn superoxide dismutase enzyme activity (ESOD), in bovines with Cu deficiency induced by high molybdenum and sulfur levels in the diet.MethodsThree similar assays were performed, with a total of twenty eight calves. The Cu-deficient group (n = 15) received a basal diet supplemented with 11 mg of Mo/kg DM as sodium molybdate, and S as sodium sulfate. The control group (n = 13) received a basal diet supplemented with 9 mg of Cu/kg DM as copper sulfate.Samples of blood and liver were taken every 28–35 days. Cu levels were measured in liver (expressed as µg/g DM), plasma (expressed as µg/dl), and erythrocytes (expressed as µg/g Hb) by flame atomic absorption spectroscopy. Superoxide dismutase (SOD1) activity was determined in red blood cells and was expressed as IU/mg hemoglobin.InfoStat Statistical Software 2020 was used for the statistical analysis. Cu levels in plasma, red blood cells and liver, and ESOD activity were analyzed by ANOVA. The correlation between erythrocyte Cu levels and the rest of the parameters were analyzed by Pearson Correlation test. Unweighted Least Squares Linear Regression of SOD1 was developed. The autocorrelation between the monthly measurements was also determined by Durbin-Watson test and autocorrelation function.ResultsThe assays lasted 314–341 days, approximately. Levels indicative of Cu deficiency for bovines were detected at 224 days (23 ± 11.6 µg/g DM) for liver Cu concentration; and at 198 days (55 ± 10.4 µg/dl) for plasma Cu concentration, in Cu-deficient animals. Liver and plasma Cu values indicative of Cu deficiency were not observed in the control group.Pearson Correlation test indicated that all indices of Cu status used in this study were significantly correlated. The highest value was obtained between ESOD and red blood Cu (0.74). There was a significant correlation between red blood Cu and plasma Cu (0.65), and with hepatic Cu (0.57). ESOD activity showed a similar significant positive correlation with liver Cu concentrations and with plasma Cu (0.59 and 0.58, respectively).ConclusionThe extremely low levels of liver and plasma Cu, the ESOD activity, erythrocyte Cu levels, and the periocular achromotrichia observed in the Cu-deficient animals showed that the clinic phase of Cu deficiency was reached in this group. The ESOD activity and erythrocyte Cu levels showed a strong association, indicating that the values of erythrocyte Cu may serve as an effective tool in assessing Cu status and diagnose a long-term Cu deficiency in cattle.     

乳腺良恶性病变剪切波弹性成像的典型表现及其鉴别诊断价值

目的:观察乳腺良恶性病变的剪切波弹性成像(SWE)的典型表现,探讨SWE对乳腺良恶性病变的鉴别诊断价值。方法:选取2017年6月～2019年6月我院收治的162例行SWE检查的乳腺肿块患者,经组织活检或病理证实良性肿块105例(良性组)、恶性肿块57例(恶性组)。对比良、恶性组SWE的典型表现、SWE参数[最大值(Emax)、最小值(Emin)、平均值(Emean)、标准差(SD)、病灶与邻近脂肪弹性比值(SWE-Ratio)]的差异,分析SWE鉴别诊断乳腺良恶性病变的价值。结果:恶性组乳腺肿块"硬边征"检出率、Ⅲ型～Ⅴ型弹性图像检出率、Emax、Emean、SD、SWE-Ratio均高于良性组(P0.05),Emin低于良性组(P0.05)。Logistic多元回归分析结果显示,"硬边征"、Emax、Emean、SWE-Ratio与病理诊断乳腺肿块性质独立相关(P0.05)。受试者工作特征(ROC)曲线分析结果显示,"硬边征"、Emax、Emean、SWE-Ratio鉴别诊断乳腺良恶性病变的曲线下面积(AUC)分别为0.923、0.686、0.873、0.879。结论:SWE是诊断乳腺良恶性病变的有效影像手段,SWE的"硬边征"、SWE-Ratio、Emean对乳腺良恶性病变具有较高的鉴别价值。     

Relative Efficiencies of Plasma Catechol Levels and Ratios for Neonatal Diagnosis of Menkes Disease

Background Menkes disease is an X-linked recessive neurodevelopmental disorder resulting from mutation in a copper-transporting ATPase gene. Menkes disease can be detected by relatively high concentrations of dopamine (DA) and its metabolites compared to norepinephrine (NE) and its metabolites, presumably because dopamine-beta-hydroxylase (DBH) requires copper as a co-factor. The relative diagnostic efficiencies of levels of catechol analytes, alone or in combination, in neonates at genetic risk of Menkes disease have been unknown. Methods Plasma from 44 at-risk neonates less than 30 days old were assayed for DA, NE, and other catechols. Of the 44, 19 were diagnosed subsequently with Menkes disease, and 25 were unaffected. Results Compared to unaffected at-risk infants, those with Menkes disease had high plasma DA (P < 10⁻⁶) and low NE (P < 10⁻⁶) levels. Considered alone, neither DA nor NE levels had perfect sensitivity, whereas the ratio of DA:NE was higher in all affected than in all unaffected subjects (P = 2 × 10⁻⁸). Analogously, levels of the DA metabolite, dihydroxyphenylacetic acid (DOPAC), and the NE metabolite, dihydroxyphenylglycol (DHPG), were imperfectly sensitive, whereas the DOPAC:DHPG ratio was higher in all affected than in all unaffected subjects (P = 2 × 10⁻⁴). Plasma dihydroxyphenylalanine (DOPA) and the ratio of epinephrine (EPI):NE levels were higher in affected than in unaffected neonates (P = 0.0015; P = 0.013). Conclusions Plasma DA:NE and DOPAC:DHPG ratios are remarkably sensitive and specific for diagnosing Menkes disease in at-risk newborns. Affected newborns also have elevated DOPA and EPI:NE ratios, which decreased DBH activity alone cannot explain.     

Schistosoma mansoni: Immunohistoperoxidase procedure in Defined Antigen Substrate Spheres (DASS) system as serologic field test

A promising serological field test for schistosomiasis could be developed by means of an immunohistoperoxidase procedure in the Defined Antigen Substrate Spheres (DASS) system. Sera of individuals with Schistosoma mansoni infections were examined with the DASS-system, using peroxidase labeled conjugates. The results were compared with those obtained with the Indirect Fluorescent Antibody (IFA) technique on frozen sections of the adult parasite. The DASS-system proved to give specific positive reactions at higher serum dilutions than the IFA-technique. The results of the test could readily be observed with the naked eye.     

A deep learning classifier for digital breast tomosynthesis

PurposeTo develop a computerized detection system for the automatic classification of the presence/absence of mass lesions in digital breast tomosynthesis (DBT) annotated exams, based on a deep convolutional neural network (DCNN).Materials and MethodsThree DCNN architectures working at image-level (DBT slice) were compared: two state-of-the-art pre-trained DCNN architectures (AlexNet and VGG19) customized through transfer learning, and one developed from scratch (DBT-DCNN). To evaluate these DCNN-based architectures we analysed their classification performance on two different datasets provided by two hospital radiology departments. DBT slice images were processed following normalization, background correction and data augmentation procedures. The accuracy, sensitivity, and area-under-the-curve (AUC) values were evaluated on both datasets, using receiver operating characteristic curves. A Grad-CAM technique was also implemented providing an indication of the lesion position in the DBT slice.Results Accuracy, sensitivity and AUC for the investigated DCNN are in-line with the best performance reported in the field. The DBT-DCNN network developed in this work showed an accuracy and a sensitivity of (90% ± 4%) and (96% ± 3%), respectively, with an AUC as good as 0.89 ± 0.04. A k-fold cross validation test (with k = 4) showed an accuracy of 94.0% ± 0.2%, and a F1-score test provided a value as good as 0.93 ± 0.03. Grad-CAM maps show high activation in correspondence of pixels within the tumour regions.Conclusions We developed a deep learning-based framework (DBT-DCNN) to classify DBT images from clinical exams. We investigated also a possible application of the Grad-CAM technique to identify the lesion position.     

Leptospirosis: Public health perspectives

Leptospirosis, caused by a spirochete of genus Leptospira, is considered the most widespread zoonosis in the world. It has a global distribution with a higher incidence in the tropics and subtropics, ranging from 10 to 100 human cases per 100,000 individuals. Leptospirosis is considered an “emerging” zoonosis due to increased contact between animals and humans and the resulting human encroachment into wildlife habitat. Climate change and its associated environmental shifts can affect the degree of transmission of leptospirosis. Surveillance for leptospirosis is important for early detection of cases because early treatment is crucial to decrease morbidity and mortality. In June 2012, the Council of State and Territorial Epidemiologists approved reinstatement of leptospirosis as a Nationally Notifiable Condition. Reinstatement of national surveillance will facilitate the assessment of the incidence, geographic distribution, trends, and risk factors associated with human cases and the identification of outbreaks and potential new animal reservoirs.