Deeper exploration of inflammatory cell populations in milk to monitor udder health in dairy cows

The present study explored the predictive values of milk leukocyte differentials (MLD) as a basis for improving the diagnosis of intramammary infections (IMIs) and subclinical mastitis. Quarter milk samples were collected for bacteriological analysis, quarter somatic cell count (qSCC), and MLD. The MLD were assessed using the cytospin technique, direct microscopic smears, and flow cytometry. The predictive values of each single leukocyte population and useful potential indices that could better reflect immune complexity were also calculated. Changes in the percentage of any leukocyte alone failed to substantially improve the predictive value of qSCC in diagnosing IMIs. Although certain parameters increased the area under the receiver operating characteristic curve (ROC curve) as a result of increased specificity values, a slight reduction in sensitivity was observed. The so-called CD8 complex was a unique parameter which improved both the sensitivity (78.79 %) and the specificity (80.77 %) in IMI diagnosis, resulting in the highest area under the ROC curve (0.87). To diagnose subclinical mastitis, the percentage of macrophages and the sum of the percentage PMNLs and T CD8+ cells divided by the percentage of macrophages showed the highest predictive values (sensitivity = 79.63, specificity = 73.68, and area under the ROC curve = 0.83) in the differentiation of the inflammatory condition status of cows. In conclusion, this study provides further insights into using T CD8+ lymphocytes in diagnosing bovine IMIs, combined with PMNLs and macrophages. The antidromic trend of macrophages vs. PMNLs and T CD8+ lymphocytes due to the increasing qSCCs was crucial to differentiating quarters under both inflammatory and non-inflammatory conditions.

The correlation between body weight, body weight gain and blood parameters in pigs at birth and weaning / A correlação entre peso corporal, ganho de peso corporal e parâmetros sanguíneos em suínos ao nascimento e desmame

The aim of study was to determine the relationship between metabolic parameters, body weight and body growth of piglets at birth and weaning. The experiment included 80 piglets obtained from F1 generation sows. Body weight was measured at birth (BW0), 24h (BW1) after birth and on day 25 (at weaning, BW2). Blood sampling was performed at the beginning of life (3rd day after birth) and at weaning (21st day after birth). BW0 and BW1 positively correlated with cholesterol and negatively with urea values at the beginning of life and RBC values at weaning. BW2 positively correlates with albumin and cortisol values at the beginning of life, total proteins, and globulins at weaning, and negatively correlates with erythrocyte values at weaning. Piglet growth from birth to weaning (BWG2-0) correlates positively with total proteins, albumin, and cortisol at the beginning of life and total proteins at weaning. ROC analysis shows that MCHC, TPROT, GLOB, CHOL and AST at the beginning of life can distinguish fast-growing from slow-growing piglets from birth to weaning period. The use of blood parameters enables early recognition of growth rate in piglets, which can help to optimize all further steps to achieve the best possible growth.

O objetivo do estudo foi determinar a relação entre parâmetros metabólicos, peso corporal e crescimento corporal de leitões ao nascimento e desmame. A experiência incluiu 80 leitões obtidos de porcas da geração F1. O peso corporal foi medido ao nascimento (BW0), 24h (BW1) após o nascimento e no dia 25 (no desmame, BW2). A amostragem de sangue foi realizada no início da vida (3º dia após o nascimento) e no desmame (21º dia após o nascimento). BW0 e BW1 correlacionaram-se positivamente com o colesterol e negativamente com os valores de uréia no início da vida e com os valores de hemácias no desmame. BW2 correlaciona-se positivamente com os valores de albumina e cortisol no início da vida, proteínas totais e globulinas no desmame, e negativamente com os valores de eritrócitos no desmame. O crescimento do leitão desde o nascimento até o desmame (BWG2-0) se correlaciona positivamente com as proteínas totais, albumina e cortisol no início da vida e com as proteínas totais no desmame. A análise ROC mostra que MCHC, TPROT, GLOB, CHOL e AST no início da vida pode distinguir leitões de crescimento rápido de leitões de crescimento lento desde o nascimento até o desmame. O uso de parâmetros sanguíneos permite o reconhecimento precoce da taxa de crescimento em leitões, o que pode ajudar a otimizar todas as etapas posteriores para alcançar o melhor crescimento possível.

CA 15-3, CRP, and LDH correlates with prognostic parameters in canine mammary neoplasms

The identification of putative prognostic factors in canine mammary neoplasms (CMNs) has been focused on tissue-specific biomarkers, but the serum biomarkers, including cancer antigen 15-3 (CA 15-3), c-reactive protein (CRP), and lactate dehydrogenase (LDH) have been demonstrated to display clinical application in cases of CMNs. The aim of the study was to evaluate the levels of these serum biomarkers and their association with well-established prognostic factors in CMNs. Samples from 15 female canines with CMNs and 15 clinically healthy ones were collected. The results were evaluated using the Tukey's, Pearson, or Spearman tests. The cut-off point, sensitivity, specificity, and area under curve (AUC) were evaluated using the receiver operating characteristic (ROC) curve analysis in a logistic regression model (P<0.05). The levels of CA 15-3, CRP and LDH were significantly higher in the serum of female dogs with CMNs compared to the healthy ones. Moreover, these factors were positively correlated with ulceration, tumor size, histopathological grade, metastatic lymph node, and clinical staging. Female dogs with CMNs were found to exhibit highest serum levels of CA 15-3, CRP, and LDH. Therefore, they can be applied to improve the efficacy of the diagnosis and prognostic evaluation in casas of CMNs.(AU)

Psychometric properties of the Brazilian version of the Canine Brief Pain Inventory (CBPI) for dogs with clinical signs of osteoarthritis and preliminary evidence of its clinical utility / Propriedades psicométricas da versão brasileira do Breve Inventário de Dor Canina (BIDC) em cães com sinais clínicos de osteoartrite e evidências preliminares da sua utilização clínica

This study aimed to determine the psychometric properties and clinical use of the Brazilian version of the Canine Brief Pain Inventory (CBPI) in dogs with hip dysplasia (HD). Forty-three dogs with HD and 16 clinically normal dogs were enrolled. The HD dogs were treated daily with 4.4mg/kg carprofen (GT = 21) or placebo (GP = 19), for four weeks. Owners completed the CBPI at two weeks (W-2) and immediately before the start of the treatment (W0), two (W2) and four (W4) weeks during treatment, and two weeks (W6) after the end of treatment. The internal structure was accessed, and the Cronbach's alpha coefficient was 0.97, indicating the high internal consistency of the instrument. Principal Component Analysis (PCA) suggested the retention of one component, which accounted for 78% of the variability. The ROC curve analysis concluded that the score 3 has an excellent performance to discriminate between normal and possible HD dogs (AUC of 0.973). There was no difference between dogs treated with carprofen versus placebo. The instrument in Portuguese showed construct and criterion validity and reliability to be used in dogs with HD.

Objetivou-se determinar as propriedades psicométricas e a utilidade clínica do Breve Inventário de Dor Canina na língua portuguesa, em cães com displasia coxofemoral (DCF). O inventário foi preenchido por tutores de 43 animais com DCF e por 16 tutores de cães saudáveis. Os animais com DCF foram tratados com carprofeno 4,4mg/kg (GT = 21) ou placebo (GP =19), administrados uma vez ao dia durante quatro semanas. As avaliações foram realizadas duas semanas e imediatamente antes do tratamento, duas e quatro semanas durante o tratamento e após duas semanas do término do tratamento. A estrutura interna calculada pelo alfa de Cronbach = 0,97 indicou alta consistência dos dados. A análise dos componentes principais identificou a retenção de apenas um componente responsável por 78% da variabilidade dos dados. A análise da curva ROC indicou que o escore 3 discrimina cães saudáveis de cães com possível DCF (ASC de 0,973). Não houve diferença entre os cães tratados com carprofeno daqueles que receberam placebo. O questionário apresentou validade de constructo e critério e confiabilidade e pode-se empregá-lo para avaliar a dor crônica em cães com osteoartrite em países de língua portuguesa.

Results of a sample-to-cutoff ratio using Abbott Architect rHTLV-I/II assay allow to predict detection of HTLV-1 and HTLV-2 proviral DNA by real-time PCR / Resultados da razão sinal da amostra sobre o cutoff da reação no ensaio Abbott Architect rHTLV-I/II permite prever a detecção de DNA proviral por PCR em tempo real

The present study aims to correlate the sample-to-cutoff ratios (S/CO) distributions of reactive results for HTLV-1/2 antibodies with the detection of proviral DNA in a population of blood donor candidates. It was carried out a retrospective data search of 632 HTLV-1/2 reactive samples, submitted to confirmatory testing from January 2015 to December 2019. Serological screening was performed by chemiluminescent microparticle immunoassay Architect rHTLV-I/II, whereas confirmatory testing was performed by in-house real-time polymerase chain reaction method. 496 out of 632 samples (78%) had undetectable HTLV-1/2 proviral DNA and 136 (22%) had detectable proviral DNA. HTLV infection was not confirmed in any individual for whom the S/CO ratio value was <4, and proviral DNA detection rates gradually escalated as S/CO ratio values increased. The sensitivity and predictive positive value found for the Architect rHTLV-I/II was 100% and 22%, respectively. The receiver operating characteristic (ROC) curve analysis showed that the optimal S/CO ratio value for predicting the presence of HTLV-1/2 was 18.11. High S/CO ratios were more associated with the detection of proviral DNA. The S/CO ratio value <4 suggests excluding true HTLV infection and the risk of blood transmission (AU).

O estudo tem como objetivo correlacionar às distribuições das razões sample-to-cutoff (S/CO) de resultados reagentes para anticorpos HTLV-1/2 com a detecção de DNA proviral em uma população de candidatos à doação de sangue. Realizou-se uma busca retrospectiva de dados de 632 amostras reagentes para HTLV-1/2 submetidas à testagem confirmatória entre janeiro de 2015 a dezembro de 2019. A triagem sorológica foi realizada pelo imunoensaio quimioluminescente de micropartículas Architect rHTLV-I/II, enquanto o teste confirmatório foi realizado pelo método de PCR em tempo real in-house. 496 de 632 amostras (78%) apresentaram DNA proviral indetectável e 136 (22%) apresentaram DNA proviral detectável. A infecção por HTLV não foi confirmada em nenhum indivíduo com valor de S/CO <4 e as taxas de detecção de DNA proviral escalonaram gradualmente à medida que as razões S/CO aumentaram. A sensibilidade e valor preditivo positivo encontrados para o Architect rHTLV-I/II foram 100% e 22%, respectivamente. Utilizando análise de curva ROC, o valor de razão S/CO ideal para predizer a presença de DNA proviral foi de 18,11. Razões S/CO elevadas foram mais associadas à detecção de DNA proviral. Em suma, o valor de S/CO <4 sugere a exclusão de infecção por HTLV e o risco de transmissão pelo sangue (AU).

Ketosis in dairy cows during early lactation - detection in pooled blood serum samples

Background: Ketosis is the most important metabolic disease with prevalence from 15 to 45%. Ketosis is diagnosed using a metabolic profile. Due to the high prevalence, it is necessary to determine a large number of metabolic profiles within farm, which represents an additional cost, so the implementation of pooled serum in assessing the metabolic status of cows was examined. The aim of this study was to validate and evaluate the influence of the relative position (Z-score) of the value of pooled sample metabolic parameters within the known reference value of healthy cows in the detection of ketosis in herd during early lactation. Materials, Methods & Results: The experiment has been carried out using blood samples collected by puncture of coccygeal vein from 50 ketotic and 50 healthy cows. Laboratory analysis includes determination of beta-hydroxybutyrate-BHB, non-esterified fatty acids-NEFA, cholesterol-CHOL, triglycerides-TGC, glucose-GLU, albumin-ALB, total protein-TPROT, UREA, Ca, P, total bilirubin-TBIL and aspartat aminotransferase-AST. The pooled serum was made from 10 individual samples originating from 10 different cows. A serum aliquot of 0.1 mL was taken from each sample, and a 1 mL volume of pooled serum was finally formed. Three types of serum pools were made: 1) 30 pooled sample were from ketosis; 2) 30 pooled sample were from healthy cows and 3) 60 pooled samples containing mixed sera of healthy cows and cows with ketosis were made as follows: 10 pools contain 10% to 60% of ketotic cows (1/10 to 6/10 samples). Statistical analysis includes: a) difference in metabolite concentration and Z-score in pooled sample and arithmetic mean individual sample in healthy and ketotic cow, b) correlation between Z-score of pooled sample and arithmetic mean of individual sample, c) ability of Z-score of metabolite to divide ketotic from healthy cow, d) correlation between Z-score and % of ketotic cow in pooled sample; and e) calculation of 95%CI of pooled sample Z-scores for each % of ketotic cow in pools. Z-score and all analysis were calculated for each metabolic parameter. The results of the study show that the mean values and Zscores of the pool and the calculated average value of the individual samples participating in that pool differ significantly in healthy cows and cows in ketosis, except for TPROT and Ca. A higher value and a higher Z-score were found for BHB, NEFA, UREA, TBIL and AST, and a lower value and a lower Z-score for TGC, CHOL, GLU, ALB and P in ketotic cows compared to healthy cows. The value of the Z-score of the pooled sample and the calculated mean values of individual samples participating in the pool are highly correlated with each other (coefficient of determination over 99%). Z-score of metabolites in the pooled sample can be used to distinguish healthy from ketotic cows (ROC AUC= 0.711 to 0.989), except for TPROT and Ca. The Z-score value of the pooled sample shows a linear correlation with the percentage of ketotic cows in the pool and the reference ranges of Z-scores change significantly as a function of the percentage of ketosis cows. Discussion: Modern research on the metabolic profile in cows requires obtaining a large amount of information from as few samples as possible. The advantages of using the Z-score are reflected in the following: this score does not depend on the absolute value of the metabolite, but on the position within the known population reference value, Z-score of sample and the arithmetic mean of individual samples included in the same pool are almost identical, the Z-score of these 2 groups of results is ideally correlated, the Z-score significantly correlates with the % of ketosis samples in the pooled sample. The use of pooled sample Z-score can be a useful in a herd level assessment of metabolic status and detection of ketosis as most important metabolic disease in dairy cows.

Effects of blood pathological changes before TAI on pregnancy of dairy cows with anestrus and estrus / [Efeitos de mudanças patológicas de plasma antes de (TAI) na gestação de vacas em anestro e estro]

The objective of this study was to investigate the influence of plasma pathological changes before timed artificial insemination (TAI) on pregnancy of cows. The contents of estrogen (E2), progesterone (P4), glucose (Glu), selenium (Se), brain-derived neurotrophic factor (BDNF), and histamine (HIS) in plasma of 48 Holstein cows were measured before TAI. According to the estrus detection, the cows were divided into estrus (E) and anestrus (A) groups. After pregnancy testing at 28 d after TAI, two groups of E and A were divided into positive pregnancy of E group (EP+), negative pregnancy of E group (EP-), positive pregnancy of A group (AP+), and negative pregnancy of A group (AP-). The contents of E2, P4, Glu, Se, BDNF and hIS significantly differed among the four groups (P<0.01). The ROC analysis was used to determine the risk of negative pregnancy test (-) after TAI was increased when plasma E2 was less than 46.45 pmol/L in cows before TAI. The changes in E2, P4,hIS, Glu, and BDNF in the blood of natural estrus and natural anestrus cows affected the pregnancy after TAI. the level of E2 in plasma may be used to assess the risk of negative pregnancy after TAI.(AU)

O objetivo do presente estudo foi investigar a influência de mudanças patológicas de plasma antes de inseminação artificial (TAI) na gestação de vacas. O conteúdo de estrogênio (E2), progesterona (P4), glucose (Glu), selênio (Se), fator neurotrófico derivado do cérebro (BDNF), e histamina (HIS) no plasma de 48 vacas Holstein foi medido antes de TAI. De acordo com a detecção de estro, as vacas foram divididas em dois grupos: estro (E) e anestro (A). Após teste de gestação 28 d após TAI, dois grupos de E e A foram formados em gestação positiva do grupo E (EP+), gestação negativa do grupo E (EP-), gestação positiva do grupo A (AP+), e gestação negativa do grupo A (AP-). Os valores de E2, P4, Glu, Se, BDNF e hIS foram significativamente diferentes entre os quatro grupos (P<0,01). A análise ROC foi utilizada para determinar o risco de teste de gestação negativo (-) após aumento de TAI quando plasma E2 estava abaixo de 46,45 pmol/L em vacas antes de TAI. Alterações em E2, P4,hIS, Glu e BDNF no sangue de estro natural e anestro natural em vacas afetou a gestação após TAI. O nível de E2 no plasma pode ser usado para avaliar o risco de gestação negativa após TAI.(AU)

Fluorescent polarization assay, two versions of indirect and competitive enzyme- linked immunoassay for brucellosis diagnosis in vaccinated buffaloes (Bubalus bubalis) / Teste de polarização fluorescente, duas versões de imunoensaio enzimático indireto e imunoensaio competitivo para diagnóstico de brucelose em búfalos vacinados (Bubalus bubalis)

ABSTRACT: The fluorescence polarization assay (FPA), two variants (V) of the indirect enzyme-linked immunosorbent assay (I-ELISA) and the competitive enzyme-linked immunosorbent assay (C-ELISA) were evaluated in buffaloes to detect antibodies against Brucella spp. The V1 of I-ELISA identifies them through the monoclonal (M23) anti-bovine IgG (I-ELISAM23) and the V2 through the ProteinA / G (I-ELISA-A/G). Serum samples of 862 buffaloes (Bubalus bubalis) from the Northeast of Argentina (NEA) were analyzed using the complement fixation test (CFT) as the reference. Receiving Operator Characteristic (ROC) analysis defined for the area under the curve (AUC) determined the cutoff points, sensitivity (Se) and specificity (Sp) for each test. CFT identified 107 positive and 755 negative sera. The best AUC (0.986), Concordance with CFT (96.3%) and kappa value (0.843) was obtained by I-ELISA A/G test. This assay showed the highest Se (95.33%) and C-ELISA the highest Sp (97%). FPA failed to measure the antibodies in 23 (2.65%) serum samples due to unsuccessful reading. I-ELISA M23 proved to be ineffective to diagnose brucellosis in bubaline sera. The four serological tests showed cutoff points lower than those standardized for bovines. As conclusion, I-ELISA A/G, C-ELISA and FPA with its limitations would be effective techniques for the diagnosis of brucellosis in buffaloes in the NEA, requiring an appropriate cut-off point to guarantee their maximum performance in this species.

RESUMO: O ensaio de polarização de fluorescência (FPA), duas variantes (V) do ensaio imunoenzimático indireto (I-ELISA) e o ensaio imunoenzimático competitivo (C-ELISA), foram avaliados em búfalos para detectar anticorpos contra Brucella spp. O V1 do I-ELISA os identifica através do IgG monoclonal (M23) anti-bovino (I-ELISAM23) e o V2 ​​através da Proteína A / G (I-ELISA-A / G). Amostras de soro de 862 búfalos (Bubalus bubalis) do Nordeste da Argentina (NEA) foram analisadas usando o teste de fixação do complemento (CFT) como referência. A análise Receiving Operator Characteristic (ROC) definida pela área sob a curva (AUC) determinou os pontos de corte, sensibilidade (Se) e especificidade (Sp) de cada teste. A CFT identificou 107 soros positivos e 755 soros negativos. Os melhores valores de AUC (0.986), concordância com CFT (96.3%) e kappa (0.843) foram obtidos pelo teste I-ELISA A / G. Este ensaio mostrou a maior Se (95.33%) e C-ELISA a maior Sp (97%). O FPA falhou em medir os anticorpos em 23 (2,65%) amostras de soro devido à falha na leitura. O I-ELISA M23 provou ser ineficaz para o diagnóstico de brucelose em soros bubalinos. Os quatro testes sorológicos mostraram pontos de corte inferiores aos padronizados para bovinos. Em conclusão, I-ELISA A / G, C-ELISA e FPA com suas limitações seriam técnicas eficazes para o diagnóstico de brucelose em búfalos no NEA, exigindo um ponto de corte adequado para garantir seu desempenho máximo nesta espécie.

Correlation of maternal concentrations of plasma testosterone with fetal sex in horses / Correlação das concentrações plasmáticas de testosterona materna com o sexo fetal em equinos

The objectives of this study were to evaluate the correlation of fetal sex and plasma testosterone concentrations between the 5th and 8th months of pregnancy in mares and to verify the applicability of this test to predict fetal sex. Blood samples were collected from 21 mares at 30-day intervals of between 150 and 240 days of pregnancy. Plasma testosterone was determined by radioimmunoassay and the sex of the foals confirmed at birth. The levels of maternal testosterone were higher in mares carrying female fetuses at months 5 and 8 (P 0.05). Limit values were determined by analyzing the receiver operating characteristic (ROC) estimates: 35.5 pg/mL and 40 pg/mL for the 5th and 8th month, respectively. For the mares with plasma testosterone values equal to or above the threshold, gestation of female foals was predicted, and for those with plasma testosterone below the threshold values pregnancy of male foals was predicted. In the 5th month, the predictive values for male and female fetuses were 70% and 88.9%, respectively; the detection rates were 87.5% and 72.7%, and the total accuracy of the examination was 78.9%. In the 8th month, the predictive values for male and female fetuses were 80% and 90%, respectively; the detection rates were 88.9% and 81.8%, and the total accuracy of the examination was 85%. It was concluded that there was a correlation between fetal sex and plasma testosterone concentrations in pregnant mares. Prediction of fetal sex based on plasma concentrations of maternal testosterone can be performed in months 5 and 8 with 78.9% and 85% accuracy, respectively.(AU)

Os objetivos do estudo foram avaliar a correlação do sexo fetal com as concentrações plasmáticas de testosterona entre o 5° e o 8º mês de gestação na égua e verificar a aplicabilidade deste exame para a predição do sexo fetal. Amostras de sangue foram coletadas de 21 éguas, com intervalos de 30 dias, entre 150 e 240 dias de gestação. A testosterona plasmática foi determinada por radioimunoensaio e o sexo dos potros foi confirmado ao nascimento. Os valores de testosterona materna foram superiores nas éguas gestando fetos fêmeas aos cinco e oito meses (P 0.05). Através da análise da curva ROC (receiver operating characteristic) foram determinados valores limites de 35,5 pg/mL e 40 pg/mL para o 5º e o 8° mês, respectivamente. Éguas com testosterona plasmática igual ou acima dos valores limites foram preditas como gestando fêmeas e éguas com testosterona plasmática abaixo dos valores limites foram preditas como gestando machos. Aos cinco meses, os valores preditivos para fetos machos e fêmeas foram 70% e 88,9%, respectivamente; as taxas de detecção foram 87,5% e 72,7% e a acurácia total do exame foi de 78,9%. Aos oito meses, os valores preditivos para fetos machos e fêmeas foram 80% e 90%, respectivamente; as taxas de detecção foram 88,9% e 81,8% e a acurácia total do exame foi de 85%. Conclui-se que houve correlação entre o sexo fetal e as concentrações de testosterona plasmática em éguas prenhes. A predição do sexo fetal baseada nas concentrações plasmáticas de testosterona materna pode ser realizada aos cinco e oito meses de gestação com 78,9% e 85% de acurácia, respectivamente.(AU)

Subclinical ketosis risk prediction in dairy cows based on prepartum metabolic indices / [Previsão do risco de cetose subclínica em vacas de leite, baseado nos índices metabólicos pré-parto]

Ketosis can seriously impair cow performance. This study detected changes in prepartum blood metabolic parameters for predicting postpartum ketosis occurrence in dairy cows. Body condition score (BCS) was assessed before and after delivery. Blood samples of 63 cows were collected from 10 days before calving to 10 days after calving to measure metabolic parameters including ß-hydroxybutyric acid (BHBA), non-esterified fatty acid (NEFA), glucose (GLU), total bilirubin (TBIL), direct bilirubin (DBIL), indirect bilirubin (IBIL), total protein (TP), albumin (ALB), globulin (GLO), alanine aminotransferase (ALT), and aspartate aminotransferase (AST). There was a postpartum subclinical ketosis incidence of 42.25%. Compared with prepartum, plasma, levels of BHBA, AST, and NEFA significantly increased postpartum, and prepartum AST (R=0.57) and NEFA (R=0.45) showed a significant positive correlation with ketosis postpartum. Plasma GLU level significantly decreased postpartum and was significantly negatively correlated with ketosis (R=-0.21). Receiver operating characteristic curve analysis revealed prepartum BSC < 2.88, and prepartum plasma AST > 68.0 U/L, GLU < 3.97mmol/L, NEFA > 0.27mmol/L, and BHBA > 0.43mmol/L, indicating a high risk of subclinical ketosis postpartum. These levels can be used as risk indicators to predict the occurrence of subclinical ketosis in postpartum cows.(AU)

A cetose pode trazer sérios prejuízos ao desempenho da vaca. Este estudo detectou alterações nos parâmetros metabólicos do sangue pré-parto para prever a cetose pós-parto que ocorre em vacas leiteiras. O escore de condição corporal (ECC) foi avaliado antes e após o parto. Foram coletadas amostras de sangue de 63 vacas entre 10 dias antes e 10 dias após o parto para medir os parâmetros metabólicos, incluindo ácido ß-hidroxibutírico (BHBA), ácido graxo não esterificado (NEFA), glicose (GLU), bilirrubina total (TBIL), bilirrubina direta (DBIL), bilirrubina indireta (IBIL), proteína total (TP), albumina (ALB), globulina (GLO), alanina aminotransferase (ALT) e aspartato aminotransferase (AST). Houve uma incidência de cetose subclínica pós-parto de 42,25%. Em comparação com o pré-parto, o plasma, os níveis de BHBA, AST e NEFA aumentaram significativamente no pós-parto, e AST no pré-parto (R = 0,57) e NEFA (R = 0,45) mostraram uma correlação significativa positiva com cetose pós-parto. O nível plasmático de GLU diminuiu significativamente no pós-parto e foi negativamente correlacionado com a cetose de forma significativa (R = -0,21). A análise da curva característica de operação do receptor revelou BSC pré-parto <2,88 e AST plasmático pré-parto> 68,0 U / L, GLU <3,97mmol / L, NEFA> 0,27mmol / L e BHBA> 0,43mmol / L, indicando um alto risco de cetose subclínica pós-parto. Esses níveis podem ser usados ​​como indicadores de risco para prever a ocorrência de cetose subclínica em vacas no pós-parto.(AU)

Fluorescent polarization assay, two versions of indirect and competitive enzyme- linked immunoassay for brucellosis diagnosis in vaccinated buffaloes (Bubalus bubalis) / Teste de polarização fluorescente, duas versões de imunoensaio enzimático indireto e imunoensaio competitivo para diagnóstico de brucelose em búfalos vacinados (Bubalus bubalis)

The fluorescence polarization assay (FPA), two variants (V) of the indirect enzyme-linked immunosorbent assay (I-ELISA) and the competitive enzyme-linked immunosorbent assay (C-ELISA) were evaluated in buffaloes to detect antibodies against Brucella spp. The V1 of I-ELISA identifies them through the monoclonal (M23) anti-bovine IgG (I-ELISAM23) and the V2 through the ProteinA / G (I-ELISA-A/G). Serum samples of 862 buffaloes (Bubalus bubalis) from the Northeast of Argentina (NEA) were analyzed using the complement fixation test (CFT) as the reference. Receiving Operator Characteristic (ROC) analysis defined for the area under the curve (AUC) determined the cutoff points, sensitivity (Se) and specificity (Sp) for each test. CFT identified 107 positive and 755 negative sera. The best AUC (0.986), Concordance with CFT (96.3%) and kappa value (0.843) was obtained by I-ELISA A/G test. This assay showed the highest Se (95.33%) and C-ELISA the highest Sp (97%). FPA failed to measure the antibodies in 23 (2.65%) serum samples due to unsuccessful reading. I-ELISA M23 proved to be ineffective to diagnose brucellosis in bubaline sera. The four serological tests showed cutoff points lower than those standardized for bovines. As conclusion, I-ELISA A/G, C-ELISA and FPA with its limitations would be effective techniques for the diagnosis of brucellosis in buffaloes in the NEA, requiring an appropriate cut-off point to guarantee their maximum performance in this species.

O ensaio de polarização de fluorescência (FPA), duas variantes (V) do ensaio imunoenzimático indireto (I-ELISA) e o ensaio imunoenzimático competitivo (C-ELISA), foram avaliados em búfalos para detectar anticorpos contra Brucella spp. O V1 do I-ELISA os identifica através do IgG monoclonal (M23) anti-bovino (I-ELISAM23) e o V2 ​​através da Proteína A / G (I-ELISA-A / G). Amostras de soro de 862 búfalos (Bubalus bubalis) do Nordeste da Argentina (NEA) foram analisadas usando o teste de fixação do complemento (CFT) como referência. A análise Receiving Operator Characteristic (ROC) definida pela área sob a curva (AUC) determinou os pontos de corte, sensibilidade (Se) e especificidade (Sp) de cada teste. A CFT identificou 107 soros positivos e 755 soros negativos. Os melhores valores de AUC (0.986), concordância com CFT (96.3%) e kappa (0.843) foram obtidos pelo teste I-ELISA A / G. Este ensaio mostrou a maior Se (95.33%) e C-ELISA a maior Sp (97%). O FPA falhou em medir os anticorpos em 23 (2,65%) amostras de soro devido à falha na leitura. O I-ELISA M23 provou ser ineficaz para o diagnóstico de brucelose em soros bubalinos. Os quatro testes sorológicos mostraram pontos de corte inferiores aos padronizados para bovinos. Em conclusão, I-ELISA A / G, C-ELISA e FPA com suas limitações seriam técnicas eficazes para o diagnóstico de brucelose em búfalos no NEA, exigindo um ponto de corte adequado para garantir seu desempenho máximo nesta espécie.

Fluorescent polarization assay, two versions of indirect and competitive enzyme- linked immunoassay for brucellosis diagnosis in vaccinated buffaloes (Bubalus bubalis) / Teste de polarização fluorescente, duas versões de imunoensaio enzimático indireto e imunoensaio competitivo para diagnóstico de brucelose em búfalos vacinados (Bubalus bubalis)

The fluorescence polarization assay (FPA), two variants (V) of the indirect enzyme-linked immunosorbent assay (I-ELISA) and the competitive enzyme-linked immunosorbent assay (C-ELISA) were evaluated in buffaloes to detect antibodies against Brucella spp. The V1 of I-ELISA identifies them through the monoclonal (M23) anti-bovine IgG (I-ELISAM23) and the V2 through the ProteinA / G (I-ELISA-A/G). Serum samples of 862 buffaloes (Bubalus bubalis) from the Northeast of Argentina (NEA) were analyzed using the complement fixation test (CFT) as the reference. Receiving Operator Characteristic (ROC) analysis defined for the area under the curve (AUC) determined the cutoff points, sensitivity (Se) and specificity (Sp) for each test. CFT identified 107 positive and 755 negative sera. The best AUC (0.986), Concordance with CFT (96.3%) and kappa value (0.843) was obtained by I-ELISA A/G test. This assay showed the highest Se (95.33%) and C-ELISA the highest Sp (97%). FPA failed to measure the antibodies in 23 (2.65%) serum samples due to unsuccessful reading. I-ELISA M23 proved to be ineffective to diagnose brucellosis in bubaline sera. The four serological tests showed cutoff points lower than those standardized for bovines. As conclusion, I-ELISA A/G, C-ELISA and FPA with its limitations would be effective techniques for the diagnosis of brucellosis in buffaloes in the NEA, requiring an appropriate cut-off point to guarantee their maximum performance in this species.(AU)

O ensaio de polarização de fluorescência (FPA), duas variantes (V) do ensaio imunoenzimático indireto (I-ELISA) e o ensaio imunoenzimático competitivo (C-ELISA), foram avaliados em búfalos para detectar anticorpos contra Brucella spp. O V1 do I-ELISA os identifica através do IgG monoclonal (M23) anti-bovino (I-ELISAM23) e o V2 ​​através da Proteína A / G (I-ELISA-A / G). Amostras de soro de 862 búfalos (Bubalus bubalis) do Nordeste da Argentina (NEA) foram analisadas usando o teste de fixação do complemento (CFT) como referência. A análise Receiving Operator Characteristic (ROC) definida pela área sob a curva (AUC) determinou os pontos de corte, sensibilidade (Se) e especificidade (Sp) de cada teste. A CFT identificou 107 soros positivos e 755 soros negativos. Os melhores valores de AUC (0.986), concordância com CFT (96.3%) e kappa (0.843) foram obtidos pelo teste I-ELISA A / G. Este ensaio mostrou a maior Se (95.33%) e C-ELISA a maior Sp (97%). O FPA falhou em medir os anticorpos em 23 (2,65%) amostras de soro devido à falha na leitura. O I-ELISA M23 provou ser ineficaz para o diagnóstico de brucelose em soros bubalinos. Os quatro testes sorológicos mostraram pontos de corte inferiores aos padronizados para bovinos. Em conclusão, I-ELISA A / G, C-ELISA e FPA com suas limitações seriam técnicas eficazes para o diagnóstico de brucelose em búfalos no NEA, exigindo um ponto de corte adequado para garantir seu desempenho máximo nesta espécie.(AU)

Diagnostic accuracy of the electrocardiogram for detection of atrial and ventricular overloads in dogs

Background: Analysis of the electrocardiogram may suggest atrial and ventricular overloads. However, it has a lowsensitivity and specificity for diagnosis of cardiac chamber overload. The accuracy of electrocardiographic interpretation can be improve using new cut-offs for the duration and amplitude of the electrocardiographic waves. Ourobjective was to evaluate the use of the electrocardiogram in the diagnosis of atrial and ventricular overload, usingechocardiography as the gold standard test for the diagnosis of atrioventricular overload. We aimed to define newcut-off values that would increase the sensitivity and specificity of the electrocardiogram for diagnosis of chamberoverload in dogs.Materials, Methods & Results: Eletrocardiogram records were obtained in 81 dogs divided into 3 groups: Group1A (healthy dogs ≤ 10 kg); Group 1B (dogs ≤ 10 kg with mitral or tricuspid valve disease); Group 2 (dogs weighingbetween 10.1 and 20 kg) and Group 3 (dogs > 20.1 kg). Duration in milliseconds (ms) and amplitude in millivolts(mV) of P waves and QRS complexes, PR and QT segment, T wave amplitude and ST segment were evaluated in leadDII. Using leads I and III, the mean cardiac electrical axis in the frontal plane, expressed in degrees, was determinedas the mean of three consecutive measurements. Values considered normal in Group 1A and 1B for the durantion ofP wave was ≤ 45 ms and QRS duration ≤ 55 ms. In Group 2 the duration of P wave was ≤ 47 ms and QRS duration ≤61 ms. In Group 3 the duration of P wave was ≤ 50 ms and duration QRS ≤ 64 ms. These values (duration of P waveand QRS duration) were compared with echocardiographic measurements of the left atrium, considering the referencevalue AE/Ao ≤ 1.4 and measurements of the left ventricle in M-mode according to the body weight, respectively. AP wave amplitude ≤ 0.4 mV suggested that the right atrium size was normal and this was...

Diagnostic accuracy of the electrocardiogram for detection of atrial and ventricular overloads in dogs

Background: Analysis of the electrocardiogram may suggest atrial and ventricular overloads. However, it has a lowsensitivity and specificity for diagnosis of cardiac chamber overload. The accuracy of electrocardiographic interpretation can be improve using new cut-offs for the duration and amplitude of the electrocardiographic waves. Ourobjective was to evaluate the use of the electrocardiogram in the diagnosis of atrial and ventricular overload, usingechocardiography as the gold standard test for the diagnosis of atrioventricular overload. We aimed to define newcut-off values that would increase the sensitivity and specificity of the electrocardiogram for diagnosis of chamberoverload in dogs.Materials, Methods & Results: Eletrocardiogram records were obtained in 81 dogs divided into 3 groups: Group1A (healthy dogs ≤ 10 kg); Group 1B (dogs ≤ 10 kg with mitral or tricuspid valve disease); Group 2 (dogs weighingbetween 10.1 and 20 kg) and Group 3 (dogs > 20.1 kg). Duration in milliseconds (ms) and amplitude in millivolts(mV) of P waves and QRS complexes, PR and QT segment, T wave amplitude and ST segment were evaluated in leadDII. Using leads I and III, the mean cardiac electrical axis in the frontal plane, expressed in degrees, was determinedas the mean of three consecutive measurements. Values considered normal in Group 1A and 1B for the durantion ofP wave was ≤ 45 ms and QRS duration ≤ 55 ms. In Group 2 the duration of P wave was ≤ 47 ms and QRS duration ≤61 ms. In Group 3 the duration of P wave was ≤ 50 ms and duration QRS ≤ 64 ms. These values (duration of P waveand QRS duration) were compared with echocardiographic measurements of the left atrium, considering the referencevalue AE/Ao ≤ 1.4 and measurements of the left ventricle in M-mode according to the body weight, respectively. AP wave amplitude ≤ 0.4 mV suggested that the right atrium size was normal and this was...(AU)

The risk assessment of oxidative stress in postpartum dairy cattle

Background: Negative energy balance in cows will induce catabolism especially of lipids and the resulting rapid increase of free fatty acids (NEFA) in the blood stream increases the production of reactive oxygen metabolites (ROMs) and promote oxidative stress. Once the animal body is in an oxidative stress state, many molecules in the body may be peroxidized, accelerating the destruction of cells and causing damage to tissues or organs. This study conducted risk assessment of the oxidative stress caused by negative postpartum energy balance in cows by exploring the relationship between negative energy balance and oxidative stress. Materials, Methods & Results: This experiment randomly selected 120 cows at 14 to 21 days postpartum, from an intensive cattle farm in Heilongjiang province, China. Using a negative energy balance standard of β-hydroxybutyric acid (BHBA) greater than 1.2 mmol/L, nonesterified fatty acid (NEFA) greater than 0.4 mmol/L and glucose (GLU) less than three mmol/L, the cattle were divided into a healthy group of 74 cows and a diseased group of 46 cows. The oxidative stress indices of the experimental cows were measured and analyzed using the independent sample t test. Spearman correlation analysis and regression analysis were performed and by using the binary Logistic regression analysis to predict disease, the receiver-operating characteristic curve (ROC) analysis established diagnosis effect and boundary values. Compared with the healthy group, the levels of glutathione peroxidase (GSH-Px), catalase (CAT), vitamin E (VE), selenium (Se) and total nitric oxide synthetase (T-NOS) in the blood of dairy cows in the diseased group were significantly reduced and the level of malondialdehyde (MDA) was significantly increased. The study concluded that negative energy balance is associated with oxidative stress in cows and the blood levels of GSH-Px, CAT and Se can be used...

Inflammatory lesions and brain tumors: is it possible to differentiate them based on texture features in magnetic resonance imaging?

Neuroimaging strategies are essential to locate, to elucidate the etiology, and to the follow up of brain disease patients. Magnetic resonance imaging (MRI) provides good cerebral soft-tissue contrast detection and diagnostic sensitivity. Inflammatory lesions and tumors are common brain diseases that may present a similar pattern of a cerebral ring enhancing lesion on MRI, and non-enhancing core (which may reflect cystic components or necrosis) leading to misdiagnosis. Texture analysis (TA) and machine learning approaches are computer-aided diagnostic tools that can be used to assist radiologists in such decisions. Methods: In this study, we combined texture features with machine learning (ML) methods aiming to differentiate brain tumors from inflammatory lesions in magnetic resonance imaging. Retrospective examination of 67 patients, with a pattern of a cerebral ring enhancing lesion, 30 with inflammatory, and 37 with tumoral lesions were selected. Three different MRI sequences and textural features were extracted using gray level co-occurrence matrix and gray level run length. All diagnoses were confirmed by histopathology, laboratorial analysis or MRI. Results: The features extracted were processed for the application of ML methods that performed the classification. T1-weighted images proved to be the best sequence for classification, in which the differentiation between inflammatory and tumoral lesions presented high accuracy (0.827), area under ROC curve (0.906), precision (0.837), and recall (0.912). Conclusion: The algorithm obtained textures capable of differentiating brain tumors from inflammatory lesions, on T1-weghted images without contrast medium using the Random Forest machine learning classifier.(AU)

Inflammatory lesions and brain tumors: is it possible to differentiate them based on texture features in magnetic resonance imaging?

Neuroimaging strategies are essential to locate, to elucidate the etiology, and to the follow up of brain disease patients. Magnetic resonance imaging (MRI) provides good cerebral soft-tissue contrast detection and diagnostic sensitivity. Inflammatory lesions and tumors are common brain diseases that may present a similar pattern of a cerebral ring enhancing lesion on MRI, and non-enhancing core (which may reflect cystic components or necrosis) leading to misdiagnosis. Texture analysis (TA) and machine learning approaches are computer-aided diagnostic tools that can be used to assist radiologists in such decisions. Methods: In this study, we combined texture features with machine learning (ML) methods aiming to differentiate brain tumors from inflammatory lesions in magnetic resonance imaging. Retrospective examination of 67 patients, with a pattern of a cerebral ring enhancing lesion, 30 with inflammatory, and 37 with tumoral lesions were selected. Three different MRI sequences and textural features were extracted using gray level co-occurrence matrix and gray level run length. All diagnoses were confirmed by histopathology, laboratorial analysis or MRI. Results: The features extracted were processed for the application of ML methods that performed the classification. T1-weighted images proved to be the best sequence for classification, in which the differentiation between inflammatory and tumoral lesions presented high accuracy (0.827), area under ROC curve (0.906), precision (0.837), and recall (0.912). Conclusion: The algorithm obtained textures capable of differentiating brain tumors from inflammatory lesions, on T1-weghted images without contrast medium using the Random Forest machine learning classifier.(AU)

Relationship between serum MMP-9 level and prognosis after radical resection for Hilar cholangiocarcinoma patients

Purpose: To analyze the preoperative serum matrix metalloproteinase-9 (MMP-9) levels and prognosis of patients with hilar cholangiocarcinoma (HC) undergoing radical resection. Methods: Preoperative serum MMP-9 levels in patients with HC undergoing radical resection were detected by enzyme-linked immunosorbent assay (ELISA). The ROC curve assay was used to analyze the preoperative serum MMP-9 level to determine the most valuable cut-off point. The relationship between MMP-9 and clinicopathological features of HC patients was analyzed. Kaplan-Meier method was used to analyze the prognostic factors, and COX regression model was used to analyze the independent risk factors affecting prognosis. Results: Preoperative serum MMP-9 levels were significantly elevated in the death patients compared with the survival patients. The most valuable cut-off point for preoperative serum MMP-9 for prognosis was 201.93 ng/mL. Preoperative serum MMP-9 was associated with Bismuth-Corlette classification) and lymph node metastasis. Kaplan-Meier analysis showed that MMP-9, Bismuth-Corlette classification, Lymph node metastasis, Portal vein invasion, Hepatic artery invasion, Liver invasion, Incised margin, and Preoperative biliary drainage were related to prognosis. Cox regression model confirmed that hepatic artery invasion, liver invasion, incised margin, and MMP-9 have the potential to independence predicate prognosis in HC patients. Conclusion: Preoperative serum MMP-9 has high predictive value for prognosis and is an independent influencing factor for the prognosis of patients with hilar cholangiocarcinoma.(AU)

Agreement between postmortem endometrial cytology, biopsy and bacteriology in culled dairy cows

The objectives of this study were to determine: a) the agreement between postmortem endometrial cytology, uterine biopsy and presence of intrauterine fluid in apparently normal genital tracts collected from culled dairy cows; b) the agreement between results obtained from paired uterine horns using the different diagnosis methods; and c) the microorganisms isolated from uteri with cytological endometritis. One hundred grossly normal genital tracts from Holstein cows were collected at a local slaughterhouse. Samples for cytology, histopathology and bacteriology were obtained from both uterine horns, and presence of any intrauterine content was determined after opening the tracts. Cut-off values used for positive cytology were ≥5% PMN (polymorphonuclear neutrophils). Data were analyzed by ROC curve analysis, Cohen’s Kappa method and Chi-square tests. Presence of inflammatory infiltrate and its endometrial distribution were the only histological parameters significantly correlated with cytology results (AUC: 0.780 and 0.694, respectively). Agreement between histology and cytology was moderately high for presence of infiltrate (kappa = 0.55) and low for infiltrate distribution (kappa = 0.33). Other histological parameters (status of endometrial epithelium, endometrial glands or fibrosis), intrauterine content or bacteriology were not correlated with cytology. Presence of intrauterine content was significantly (P = 0.036) related with biopsy results. Moderately high agreement was observed between paired uterine horns for cytology (kappa = 0.65) and biopsy (kappa = 0.74). Bacteriology was not related with cytology or biopsy, but different bacteria were isolated from cytology negative or positive uteri. It was concluded that endometrial cytology, although less sensitive than biopsy, is a useful tool for diagnosing subclinical endometrial inflammation.

Agreement between postmortem endometrial cytology, biopsy and bacteriology in culled dairy cows

The objectives of this study were to determine: a) the agreement between postmortem endometrial cytology, uterine biopsy and presence of intrauterine fluid in apparently normal genital tracts collected from culled dairy cows; b) the agreement between results obtained from paired uterine horns using the different diagnosis methods; and c) the microorganisms isolated from uteri with cytological endometritis. One hundred grossly normal genital tracts from Holstein cows were collected at a local slaughterhouse. Samples for cytology, histopathology and bacteriology were obtained from both uterine horns, and presence of any intrauterine content was determined after opening the tracts. Cut-off values used for positive cytology were ≥5% PMN (polymorphonuclear neutrophils). Data were analyzed by ROC curve analysis, Cohens Kappa method and Chi-square tests. Presence of inflammatory infiltrate and its endometrial distribution were the only histological parameters significantly correlated with cytology results (AUC: 0.780 and 0.694, respectively). Agreement between histology and cytology was moderately high for presence of infiltrate (kappa = 0.55) and low for infiltrate distribution (kappa = 0.33). Other histological parameters (status of endometrial epithelium, endometrial glands or fibrosis), intrauterine content or bacteriology were not correlated with cytology. Presence of intrauterine content was significantly (P = 0.036) related with biopsy results. Moderately high agreement was observed between paired uterine horns for cytology (kappa = 0.65) and biopsy (kappa = 0.74). Bacteriology was not related with cytology or biopsy, but different bacteria were isolated from cytology negative or positive uteri. It was concluded that endometrial cytology, although less sensitive than biopsy, is a useful tool for diagnosing subclinical endometrial inflammation.(AU)