Study on prevalence and liver function test enzymes of differently plumaged peafowls (Pavo cristatus) infected with Toxoplasma gondii in captivity / [Estudo sobre enzimas de prevalência e teste da função hepatica de pavões de plumage diferente (Pavo cristatus) infectados com Toxoplasma gondii em cativeiro]

O presente estudo foi realizado para determinar a prevalência geral de toxoplasmose em pavões de plumagem diferente e seu efeito nas enzimas de teste da função hepática dos hospedeiros. Um total de cem pavões de plumas diferenciais, como ombro preto (n = 52), azul (n = 28), branco (n = 10) e arlequim (n = 10) foram estudados no zoológico de Bahawalpur, no Paquistão, usando o Latex Agglutination Test (LAT) e ensaio imunossorvente ligado a enzima (ELISA). A prevalência geral por LAT e ELISA foi de 37% e 30%, respectivamente. Por LAT, observou-se uma prevalência não significativamente maior (P≥0,05) em gênero (37,77%) nos machos do que nas fêmeas (36,36%), enquanto os adultos apresentaram uma prevalência maior (37,97%) em relação aos jovens (33,33%). De acordo com o ELISA, uma prevalência significativamente (P <0,05) maior (35,55%) foi observada nos machos do que nas fêmeas (25,45%) e significativamente (P <0,05) maior prevalência (31,64%) foi registrada nos adultos do que nos jovens (23,80%). A análise do perfil bioquímico sérico mostrou que o nível de bilirrubina não teve elevação significativa nos hospedeiros infectados, em comparação aos não infectados, enquanto a concentração de albumina, alanina aminotransferase (ALT), aspartato aminotransferase (AST), fosfatase alcalina (ALP) foi significativamente (P <0,05) diferente nos hospedeiros infectados. Conclui-se que a toxoplasmose afeta as enzimas do teste da função hepática. Essa é uma pesquisa preliminar e requer mais pesquisas em todo o país, com populações e amostras maiores.(AU)

Avaliação hematológica e hepática de ovinos sob tratamento com torta de neem (Azadirachta indica) / Hematological and liver evaluation of sheep under treatment with neem pie (Azadirachta indica)

Devido aos altos índices de resistência dos parasitas aos diferentes princípios ativos comerciais, novas alternativas de controle vêm sendo estudadas, entre elas a fitoterapia. Essas medidas visam a busca de métodos auxiliares no controle das parasitoses, entretanto, muitos produtos estão disponíveis no mercado e não têm comprovação científica de sua eficácia ou de possíveis efeitos colaterais. O objetivo deste estudo foi avaliar os efeitos hematológicos e hepáticos após a administração de torta de Neem (Azadirachta indica) em ovinos. Foram testadas três dosagens da torta de Neem adicionada ao sal mineral (1, 2 e 4%), administradas por 126 dias para 32 ovinos da raça Lacaune, divididos em quatro grupos sendo três grupos para os diferentes tratamentos e um controle, o qual recebeu somente sal mineral. Amostras de sangue foram colhidas a cada 21 dias para realização do hemograma completo, dosagem de proteína plasmática total e fibrinogênio e avaliação da bioquímica clínica hepática. Nestas mesmas ocasiões, amostras de fezes foram coletadas para a quantificação de ovos por grama de fezes (OPG). Foram observadas diferenças estatísticas entre momentos e grupos para diversas variáveis, porém sem estarem relacionadas à administração de torta de Neem. Os resultados obtidos de hemograma completo, dosagem de proteína plasmática total, fibrinogênio e de bioquímica clínica hepática indicaram que a administração de torta de Neem nas concentrações de 1, 2 e 4%, não interfere nos valores hematológicos, nem sobre a integridade e função hepática de ovinos da raça Lacaune.

Due to high levels of parasite resistance to different commercial active ingredients, new control alternatives are being studied, including the phytotherapy. These measures aim to search for auxiliary methods in the control of parasitic diseases. However, there are many products available in the market and there are no scientific proof of its efficacy. The objective of this study was to evaluate the hematological and hepatic effects following administration of Neem pie (Azadirachta indica) in sheep. Three concentrations of Neem cake was added to the mineral salt (1, 2 and 4%) and administered during 126 days to 32 Lacaune breed sheeps, divided into four groups: three groups for different treatments and a and a control were tested, the ladder receiving only mineral salt. Blood samples were taken every 21 days to perform the complete blood count, serum total plasma protein and fibrinogen and liver biochemical evaluation. In those same times, fecal samples were collected for quantification of eggs per gram of feces (EPG). Statistical differences between times and groups for several variables were observed, but without being related to the administration of Neem pie. The results of complete blood count, measurement of total plasma protein, fibrinogen and hepatic clinical biochemistry indicated that administration of Neem pie at concentrations of 1, 2 and 4%, does not interfere in hematological values, or on the integrity and liver function Lacaune sheep breed.

Blood parameters and hepatic histopathology of broilers fed rations supplemented with essential oils

This study aimed to evaluate the hematological profile, hepatic function, and histopathology of mixed-sex broilers fed rations supplemented with microencapsulated essential oils from Cymbopogon flexuosus (lemon grass) and Lippia rotundifolia (chá-de-pedestre). One hundred and fifty Cobb chicks were housed in cages from 1 to 42 days of age in a completely randomized design, with six replicates with five chickens in each of five treatments (150 total chicks): basal diet without antibiotic growth promoter (negative control), diet with enramicina and salinomycin (positive control), diet with lemon grass essential oil, diet with L. rotundifolia essential oil, and diet with a mixture of lemon grass and L. rotundifolia essential oils. Mean corpuscular volume (MCV) and mean corpuscular hemoglobin (MCH) were significantly lower in untreated broilers (negative control group). Gender was not associated with erythrogram values, but aspartate aminotransferase activity (AST) was higher in females. Gamma-glutamyl transferase (GGT) was higher in chicks in the group that received rations with lemon grass oil. Broilers that received L. rotundifolia oil developed more hepatic lesions, although no effect of sex was observed related to the lesion score. Biliary hyperplasia and fibroplasias were observed in all groups, with higher histopathology scores in broilers that received diets containing L. rotundifolia oil. Mixed-sex broilers fed rations supplemented with lemon grass and L. rotundifolia essential oils have normal complete blood counts and unspecific hepatic lesions and are characterized by lipidosis, hyperplasia of the bile ducts, and fibroplasia.(AU)

Relationship among blood indicators of hepatic function and lipid content in the liver during transitional period in high-yielding dairy cows

Background: Production diseases, such as those associated with improper nutrition or management are common in transitional high-yielding dairy cows. The diseases listed in this include: the fat liver syndrome, ketosis, oxidative stress, laminitis, mastitis, milk fever, retained placenta, metritis and infertility. The diseases occur mainly around calving. They are all interrelated and form the so-called periparturient disease complex. The term transition is to underscore the important physiological, metabolic, nutritional and endocrine changes occurring in this time frame. Diagnosing liver lipidosis and susceptibility of ketosis in dairy cows may include liver biopsy or ecography, but a less invasive and more economical analytical method may be the measurement of blood biochemical metabolites. The objective of the present study was to determine a relationship among blood indicators of hepatic function and lipid content in the liver during transitional period in high-yielding dairy cows. Materials, Methods & Results: Late pregnant and calved cows (n = 40) were selected from a Holstein dairy herd and allocated to four groups: a late pregnant cows (n = 10) from day 15 to day 5 before calving; late pregnant cows (n = 10) from day 4 to day 1 before calving; clinically puerperal healthy cows (n = 10) and clinically ketotic puerperal cows (n = 10). Liver and blood samples were taken from all cows. The blood metabolites concentrations were determined by photometric methods using a Cobas Mira automatic analyzer. Liver specimens were histopathologically analyzed for lipid contents using a freezing microtome. The statistical analysis of the obtained data was carried out by ANOVA-procedure. The results of present investigation have shown that the lipid content in the liver and the blood non-esterificed fatty acids, ß-hydroxybutyrate, total bilirubin concentrations and the AST activities were significantly higher (P < 0.05) as well the blood glucose, total cholesterol, triglycerides and albumin concentrations were significantly lower (P < 0.05) in puerperal ketotic cows as compared to the values of these parameters in the blood of healthy cows in the transition period. Discussion: Liver biopsy is the only reliable method to determine severity of fatty liver in dairy cattle in the transitional period. Blood, urine and milk metabolites or blood enzyme activity have been proposed as diagnostic tools. This investigation demonstrated that in healthy transitional cows a mild fatty infiltration occurred in liver during the late pregnancy and early lactation. The histopathological examination showed a moderate to severe degree of fatty liver in ketotic cows. The lipomobilisation markers, serum BHB and NEFA concentrations, were markedly enhanced in puerperal ketotic cows. However, liver steatosis compromised hepatocyte metabolism, leading to signifi cantly weaker circulating concentrations of glucose, TG and total cholesterol, and induced some cellular lesions as evidenced by significant increases in the serum bilirubin concentrations and in the AST enzyme activities in puerperal ketotic cows. All these biochemical metabolites may be used as important biochemical indicators in the determination of the functional status of the liver in high-yielding dairy cows during the transition period.

Clonagem do cDNA e sequenciamento parcial do gene que codifica a enzima glutamato desidrogenase hepática de ovino / Cloning of the cDNA and partial sequencing of the gene that codifi es the sheep hepatic enzyme glutamate dehydrogenase

A enzima mitocondrial glutamato desidrogenase (GDH: EC 1.4.1.2) catalisa a desaminação reversível do L-glutamato para 2-oxoglutarato (α-cetoglutarato) usando o NAD+ e NADP+ como coenzimas. É uma das mais importantes enzimas hepáticas encontradas em hepatócitos de bovinos, ovinos e caprinos. Infecções por Fasciola spp., intoxicação grave aguda por toxinas de plantas, tais como Xanthium spp. e Senecio spp. e intoxicação por cobre resultam na liberação dessa enzima no sangue. O aumento da GDH indica danos ou necrose hepática em bovinos e ovinos. Esta é a enzima de escolha para avaliar a função hepática dos ruminantes. No presente trabalho o cDNA que codifica a enzima GDH do hepatócito de ovino foi sintetizado por meio de RT-PCR utilizando mRNA extraído do fígado de ovino. Parte da região de codificação do cDNA da GDH de ovino foi amplificada por PCR usando oligonucleotídeos iniciadores sintetizados a partir do alinhamento de sequências de ORFs de Ovis aries, Bos taurus, Homo sapiens, Rattus norvegicus e Mus musculus disponíveis em banco de dados. O cDNA foi clonado no vetor pGEM® -T Easy (Promega) e inserido em células cálcio-competentes de Escherichia coli DH10B através de choque térmico. O DNA plasmidial foi purificado e após o sequenciamento a presença de um inserto de 1292 pb foi confirmado. O alinhamento da sequência deduzida de aminoácidos com outras espécies revelou alta homologia entre as GDH.(AU)

The mitochondrial enzyme Glutamate Dehydrogenase (GDH: EC 1.4.1.2) catalyzes the reversible deamination of the L-glutamate for 2-oxoglutarate (α-ketoglutarate) using NAD+ and NADP+ as coenzymes. It is one of the most important liver enzymes found in hepatocytes of cattle, sheep and goats. Infections by Fasciola spp., severe acute intoxication by toxins of plants such as Xanthium spp. and Senecio spp. as well as intoxication by copper result in the release of this enzyme in blood. The increase of the GDH indicates damage or hepatic necrosis in cattle and sheep. This is an enzyme of choice to evaluate the function of the ruminants. In the present study the cDNA, that codifies the GDH enzyme of the hepatocyte of sheep, was synthesized by means of RT-PCR making use of mRNA extracted from the liver of sheep. Part of the region where the cDNA of the GDH of the ovine is codified was amplified by PCR from primers synthesized through the comparison of the aligned sequences of Ovis aries, Bos taurus, Homo sapiens, Rattus norvegicus and Mus musculus available in the database. The cDNA was cloned in the vector pGEM®-T Easy (Promega) and inserted in Escherichia coli DH10B calcium competent cells by heat shock procedure. The plasmid DNA was purified and after sequencing, the presence of 1292 pb was confirmed. The alignment of the sequence deduced of amino acid with other species revealed high homology among the GDHs.(AU)

Clonagem do cDNA e sequenciamento parcial do gene que codifica a enzima glutamato desidrogenase hepática de ovino / Cloning of the cDNA and partial sequencing of the gene that codifi es the sheep hepatic enzyme glutamate dehydrogenase

A enzima mitocondrial glutamato desidrogenase (GDH: EC 1.4.1.2) catalisa a desaminação reversível do L-glutamato para 2-oxoglutarato (α-cetoglutarato) usando o NAD+ e NADP+ como coenzimas. É uma das mais importantes enzimas hepáticas encontradas em hepatócitos de bovinos, ovinos e caprinos. Infecções por Fasciola spp., intoxicação grave aguda por toxinas de plantas, tais como Xanthium spp. e Senecio spp. e intoxicação por cobre resultam na liberação dessa enzima no sangue. O aumento da GDH indica danos ou necrose hepática em bovinos e ovinos. Esta é a enzima de escolha para avaliar a função hepática dos ruminantes. No presente trabalho o cDNA que codifica a enzima GDH do hepatócito de ovino foi sintetizado por meio de RT-PCR utilizando mRNA extraído do fígado de ovino. Parte da região de codificação do cDNA da GDH de ovino foi amplificada por PCR usando oligonucleotídeos iniciadores sintetizados a partir do alinhamento de sequências de ORFs de Ovis aries, Bos taurus, Homo sapiens, Rattus norvegicus e Mus musculus disponíveis em banco de dados. O cDNA foi clonado no vetor pGEM® -T Easy (Promega) e inserido em células cálcio-competentes de Escherichia coli DH10B através de choque térmico. O DNA plasmidial foi purificado e após o sequenciamento a presença de um inserto de 1292 pb foi confirmado. O alinhamento da sequência deduzida de aminoácidos com outras espécies revelou alta homologia entre as GDH.

The mitochondrial enzyme Glutamate Dehydrogenase (GDH: EC 1.4.1.2) catalyzes the reversible deamination of the L-glutamate for 2-oxoglutarate (α-ketoglutarate) using NAD+ and NADP+ as coenzymes. It is one of the most important liver enzymes found in hepatocytes of cattle, sheep and goats. Infections by Fasciola spp., severe acute intoxication by toxins of plants such as Xanthium spp. and Senecio spp. as well as intoxication by copper result in the release of this enzyme in blood. The increase of the GDH indicates damage or hepatic necrosis in cattle and sheep. This is an enzyme of choice to evaluate the function of the ruminants. In the present study the cDNA, that codifies the GDH enzyme of the hepatocyte of sheep, was synthesized by means of RT-PCR making use of mRNA extracted from the liver of sheep. Part of the region where the cDNA of the GDH of the ovine is codified was amplified by PCR from primers synthesized through the comparison of the aligned sequences of Ovis aries, Bos taurus, Homo sapiens, Rattus norvegicus and Mus musculus available in the database. The cDNA was cloned in the vector pGEM®-T Easy (Promega) and inserted in Escherichia coli DH10B calcium competent cells by heat shock procedure. The plasmid DNA was purified and after sequencing, the presence of 1292 pb was confirmed. The alignment of the sequence deduced of amino acid with other species revealed high homology among the GDHs.

Relationship among blood indicators of lipomobilization and hepatic function during early lactation in high-yielding dairy cows.

Blood indicators are used as a tool to diagnose metabolic disorders. The present work was conducted to study the relationships among blood indicators of lipomobilization and hepatic function in high-yielding dairy cows. Two groups of Holstein cows were studied: 27 early lactation cows and 14 mid lactation cows from four different herds with similar husbandry characteristics in Galicia, Spain. Blood samples were obtained to measure beta-hydroxybutyrate (BHB), non-esterified fatty acids (NEFA), triglycerides (TG), and the activity of aspartate transaminase (AST) and gamma-glutamyl transferase. Cows in early lactation had higher levels of BHB and NEFA than mid lactation cows. High lipomobilization (NEFA > 400 µmol/L) was detected in 67% and 7% of early lactation and mid lactation cows, respectively, while subclinical ketosis (BHB > 1.2 mmol/L) was detected in 41% and 28% of the early lactation and lactation cows, respectively. TG concentrations were low in all cows suffering subclinical ketosis and in 61% of the cows with high lipomobilization. During early lactation, 30% of cows suffered hepatic lipidosis as detected by levels of AST. Compromised hepatic function was observed in early lactation cows as shown by lower concentrations of glucose, total protein, and urea.

Valores de referência e influência do fator etário sobre os parâmetros bioquímicos utilizados para avaliação da função hepática de bubalinos hígidos da raça Murrah / Reference values and influence of age in biochemistry of liver function in healthy Murrah buffaloes

Para o estabelecimento de valores bioquímicos de referência e verificação da influência do fator etário sobre os parâmetros bioquímicos para avaliação da função hepática de bubalinos hígidos da raça Murrah, utilizaram-se amostras de soro sanguíneo de 125 animais, distribuídos em 04 grupos experimentais, de acordo com a idade: 0 a 3 meses de idade (Grupo 1 - G1), 4 a 6 meses de idade (Grupo 2 - G2); 7 a 12 meses de idade (Grupo 3 G3), e com idade superior a 01 ano (Grupo 4 G4). Os constituintes bioquímicos estudados foram: enzimas gama glutamiltransferase (GGT) e aspartato aminotransferase (AST), além da verificação dos teores séricos da proteína total (PT), da albumina e das globulinas. As atividades das enzimas GGT e AST foram determinadas através da utilização de kits cinéticos ultravioletas e a leitura realizada em aparelho analisador bioquímico semi-automático. Os valores da PT e albumina foram determinados através da utilização de métodos colorimétricos e a leitura foi realizada em espectrofotômetro. O teor sérico das globulinas foi determinado pela subtração da albumina sobre os valores séricos de PT. A atividade da GGT foi de 35,9; 19,6; 25,1; e 33,3 UI/L, respectivamente, nos animais dos grupos 1, 2, 3 e 4, sendo significativamente maior nos G1 e G4. A atividade da AST menor nos primeiros meses de vida - G1 (92,3 UI/L), aumentando para 128, 177 e 143 UIL nos animais dos grupos 2, 3 e 4, respectivamente. Os valores médios encontrados foram de 8,2; 7,0; 6,9; 7,7 g/dL de PT; 2,4; 2,2; 2,1; e 2,5 g/dL de albumina; e 5,8; 4,7; 4,7; e 5,1 g/dL de globulinas, respectivamente, nos animais dos grupos 1, 2, 3 e 4. Com base nos resultados obtidos, foi possível verificar a influência dos fatores etários nos parâmetros avaliados.

In order to determine biochemical reference values and to assess the influence of age in the biochemistry of liver function in healthy Murrah buffaloes, serum samples of 125 animals were analyzed. Buffaloes were divided into four experimental groups according to age: 0 to 3 months (Group 1 - G1), 4 to 6 months (Group 2 - G2), 7 to 12 months (Group 3 G3), and over 01 year of age (Group 4 G4). The following biochemical parameters were analyzed: enzymes gamma glutamyl transferase (GGT) and aspartate aminotransferase (AST), total serum protein (PT), albumin and globulin. GGT and AST activities were determined by means of UV assays and reading was carried out in a semi-automatic biochemistry analyzer. PT and albumin were determined by colorimetric methods and reading was carried out in a spectrophotometer. Serum globulin concentration was determined by subtracting albumin from total protein concentration. GGT activity was equal to 35.9; 19.6; 25.1; and 33.3 IU/L in animals of Groups 1, 2, 3 and 4, respectively, and was significantly greater in G1 and G4. AST activity was lower in the first months of life- G1 (92.3 IU/L), increasing to 128, 177 and 143 IU/L in animals of Groups 2, 3 and 4, respectively. Mean values recorded for PT were 8.2; 7.0; 6.9; 7.7 g/dL; albumin, 2.4; 2.2; 2.1; and 2.5 g/dL; and globulins, 5.8; 4.7; 4.7; and 5.1 g/dL in animals of Groups 1, 2, 3 and 4, respectively. Based on these results it was possible to assess the influence of age in the parameters studied.

Intoxication of Crotalaria pallida seeds to growing broiler chicks.

The adverse effects of feeding Crotalaria pallida (CP) seeds to chicks was investigated in a 21-day randomized trial of 4 dietary treatments (control, 1,2 and 3% ground CP seeds). Mortality rates in birds fed 0, 1, 2, and 3% dietary CP were 0, 2.1, 6.2, and 16.7%, respectively. Body weight gain and feed efficiency were adversely affected by all levels of inclusion of CP seeds, but feed intake was decreased only by dietary levels of 2 and 3%. Dietary CP of 2 and 3% increased the relative weight of lung, heart and spleen. Relative liver weight was increased by 2% dietary CP, but decreased by 3% CP. At day 14, serum GGT was increased by 2 and 3% dietary CP; serum ALT was significantly increased by 3% CP. No differences in ALT, AST or GGT were observed at day 21. Dietary levels equal to or greater than 1% CP are toxic for growing broiler chicks.