Hepatic lipid accumulation is associated with multiple metabolic pathway alterations but not dyslipidemia and insulin resistance in central bearded dragons (Pogona vitticeps).

OBJECTIVE: To investigate associations between hepatic fat accumulation, fibrosis, and plasma values of primary metabolites, biochemical measurands, insulin, and lipoproteins in bearded dragons. ANIMALS: 48 adult central bearded dragons (Pogona vitticeps). METHODS: Dragons were sedated with alfaxalone, and a blood sample was collected. Plasma was submitted for untargeted primary metabolomics using gas chromatography time-of-flight mass spectrometry, a biochemistry panel, and a lipoprotein panel determined by PAGE. Hepatic lipid content was quantified by liver attenuation measurements from CT images and digital image analysis of standardized histologic sections of the liver. Fibrosis was quantified by digital image analysis on Masson's trichrome-stained histologic sections. Severity was determined from pathologic review of liver sections according to a standardized grading system. Statistical associations were investigated using serial linear models adjusted for false discovery rate and multivariate statistics. RESULTS: Both hepatic fat and fibrosis had a significant effect on CT liver attenuation values. Several oligosaccharides (maltotriose, maltose, ribose, trehalose) and alkaline phosphatase were significantly and linearly increased with hepatic lipid content (all q < .05). On partial least square-discriminant analysis, ß-hydroxybutyric acid was the most important discriminatory variable between fatty liver severity grades on histology. No significant associations were found with insulin, lipoproteins, and succinic acid. CLINICAL RELEVANCE: Bearded dragons with hepatic lipid accumulation experienced multiple metabolic pathway disruptions, some being compatible with mitochondrial dysfunction. No evidence of insulin resistance or dyslipidemia was found. Hepatic biopsy and histopathology remain recommended for reliably diagnosing and staging fatty liver disease in bearded dragons.

Climbazole causes cell apoptosis and lipidosis in the liver of grass carp.

Climbazole, an azole, is widely used in personal care products, pharmaceuticals, and pesticides and is frequently detected in surface water. Climbazole has showed endocrine-disrupting effects. However, the effects of climbazole in fish are still largely unclear. In this study, grass carp (Ctenopharyngodon idella) and liver cell lines (L8824 cells) were treated with climbazole at concentrations ranging from 0.2 to 20 µg/L for 42 days in vivo and 24 h in vitro to evaluate the effects on the liver, respectively. Pathological, biochemical, and gene transcription and expression analyses were conducted to examine the hepatotoxicity. Our results showed that climbazole significantly decreased the hepatosomatic index, caused cell apoptosis in vivo and in vitro, and finally accumulated lipids in the liver. Beside, climbazole increased ROS levels, reduced Nrf2 and Keap1 mRNA and protein levels, and further decreased transcription of Nrf2-dependent downstream antioxidant enzyme genes, causing oxidative stress. Moreover, climbazole increased transcription and protein levels of apoptosis-related genes. Finally, climbazole damaged mitochondrial function and structure, disrupted liver lipid metabolism. Overall, climbazole caused hepatotoxicity, leading to a high ecological risk for aquatic organisms.

Rumen-protected choline reduces hepatic lipidosis by increasing hepatic triacylglycerol-rich lipoprotein secretion in dairy cows.

Objectives were to determine the effects of supplementing rumen-protected choline (RPC) on hepatic composition and secretion of triacylglycerol-rich lipoprotein when cows were subjected to feed restriction to develop fatty liver. It was hypothesized that RPC reduces hepatic triacylglycerol by enhancing secretion of hepatic lipoprotein. Pregnant, nonlactating parous Holstein cows (n = 33) at mean (± standard deviation) 234 ± 2.2 d of gestation were blocked by body condition (3.79 ± 0.49) and assigned to receive 0 g/d (CON), 25.8 g/d choline ion from a RPC product containing 28.8% choline chloride (CC; treatment L25.8), or 25.8 g/d of choline ion from a RPC product containing 60.0% CC (H25.8). Cows were fed for ad libitum intake for the first 5 d and restricted to 41% of the net energy for lactation required for maintenance and pregnancy from d 6 to 13. Intake of metabolizable methionine was maintained at 18 g/d during feed restriction by supplying rumen-protected methionine. Hepatic tissue was sampled on d 6 and 13 and analyzed for triacylglycerol and glycogen, and mRNA expression of hepatic tissue was investigated. On d 14, cows were not fed and received a 10% solution of tyloxapol intravenously at 120 mg/kg of body weight to block hydrolysis of triacylglycerols in very low density lipoprotein (VLDL). Blood was sampled sequentially for 720 min and analyzed for concentration of triacylglycerol and total cholesterol. Lymph was sampled 6 h after tyloxapol infusion, and analyzed for concentrations of fatty acids, ß-hydroxybutyrate, glucose, triacylglycerol, and total cholesterol. A sample of serum collected at 720 min after tyloxapol was assayed for the metabolome composition. The area under the curve (AUC) of serum triacylglycerol, VLDL cholesterol, and total cholesterol were calculated. Orthogonal contrasts evaluated the effect of supplementing RPC (CON vs. [1/2 L25.8 + 1/2 H25.8]) and source of RPC (L25.8 vs. H25.8). Least squares means and standard errors of the means are presented in sequence as CON, L25.8, H25.8. During feed restriction, supplementation of RPC reduced hepatic triacylglycerol (9.0 vs. 4.1 vs. 4.5 ± 0.6%) and increased glycogen contents (1.9 vs. 3.5 vs. 4.1 ± 0.2%). Similarly, supplementation of RPC increased the expression of transcripts involved in the synthesis and assembly of lipoproteins (MTTP), cellular autophagy (ATG3), and inflammation (TNFA), and reduced the expression of transcripts associated with mitochondrial oxidation of fatty acids (HADHA, MLYCD) and stabilization of lipid droplets (PLIN2). After infusion of tyloxapol, RPC increased the AUC for serum triacylglycerol (21,741 vs. 32,323 vs. 28,699 ± 3,706 mg/dL × min) and VLDL cholesterol (4,348 vs. 6,465 vs. 5,740 ± 741 mg/dL × min) but tended to reduce the concentrations of triacylglycerol in lymph (16.7 vs. 13.8 vs. 11.9 ± 1.9 mg/dL). Feeding RPC tended to increase the concentrations of 89 metabolites in serum, after adjusting for false discovery, including 3 acylcarnitines, 1 AA-related metabolite, 11 bile acids, 1 ceramide, 6 diacylglycerols, 2 dihydroceramides, 1 glycerophospholipid, and 64 triacylglycerols compared with CON. Feeding 25.8 g/d of choline ion as RPC mediated increased hepatic triacylglycerol secretion to promote lipotropic effects that reduced hepatic lipidosis in dairy cows.

Hepatic Lipidosis in Ruminants.

Hepatic lipidosis (ie, fatty liver) occurs primarily during the first weeks of lactation in dairy cows because of excessive lipolysis overwhelming the concomitant capacity for beta-oxidation and hepatic export of triglycerides. Besides economic losses due to reduced lactational and reproductive performance, close associations with concomitantly occurring infectious and metabolic health disorders, in particular ketosis, exist. Hepatic lipidosis is not only a consequence from the postpartal negative energy balance but also acts as a disease component for further health disorders.

Morphological features of hepatic lipid changes in bearded dragons (Pogona vitticeps), and a proposed grading system.

Hepatic lipidosis is commonly diagnosed in pet bearded dragons (Pogona vitticeps). However, there are no studies detailing the histological features of hepatic lipid changes in this species. This study describes the microscopic features of lipid change and details an associated scoring system. Histologic hepatic sections were retrospectively evaluated from 252 bearded dragons submitted for necropsy. Pathologic assessment was used to develop a grading scheme with 2 qualitative, 1 quantitative, and 6 semi-quantitative microscopic parameters, which were refined based on variability. The final grading system developed for diffuse and panlobular lipid accumulation included 2 semi-quantitative and 1 quantitative categories: percentage of hepatocellular vacuolation, fibrosis, and hepatocellular swelling, respectively. Hepatocellular swelling was indirectly quantified by counting the number of nuclei per unit area. There was a strong positive correlation (P < .001) between the percentage of hepatocellular vacuolation and lipid content, a strong negative correlation (P < .001) between nuclear count and lipid content, and a moderate correlation (P < .001) between fibrosis and lipid content. Each category was given a numerical value ranging from 0 to 4, with the sum of each representing the final grade. Cutoff values stratified microscopic changes into mild (final grade 1-4), moderate (5-7), and severe (≥8). There was strong interrater agreement for assessment of vacuolization, fibrosis, and severity classification and moderate for hepatocellular swelling. This study documents the features of hepatic lipid changes in bearded dragons. Although a cutoff to differentiate pathologic from nonpathologic lipid accumulation could not be estimated, the proposed grading scheme can be used to inform future studies.

Hemato-biochemical and ultrasonographic evaluation of hepatic lipidosis in dairy buffaloes.

Around 60% dairy animals developed moderate to severe hepatic lipidosis at the time of parturition or during early lactation stage. Most of clinician suspect the hepatic lipidosis during above time window only. However, negative energy balance or feeding of high concentrate diet can lead to hepatic lipidosis at any phase of life. The aim of the present study was to evaluate the potential for diagnosis of hepatic lipidosis by means of hemato-biochemical parameters and ultrasonography of the liver at any stage of life. Here, ultrasonographic back fat thickness measurement was correlated with ultrasonographic features of hepatic lipidosis. A total 60 buffaloes were included under the study and sampled for hematological and biochemical parameters. Hematological parameters did not exhibit any significant difference between healthy and hepatic lipidosis-affected buffaloes. Biochemical parameters like beta hydroxy butyric acid, non esterified fatty acid, aspartate amino transferase, gamma glutamyl transferase and alkaline phosphatase revealed a significant increase, while triglyceride, cholesterol, and glucose declined significantly in hepatic lipidosis-affected buffaloes. Total protein, albumin, and total bilirubin levels did not exhibit any significant difference. Based on ultrasonographic findings, the hepatic lipidosis-affected buffaloes were further sub divided into mild, moderate, and severe groups. Portal vein diameter and depth of portal vein were also estimated in current study. Ultrasonographic examination could diagnose 53.33% hepatic lipidosis cases in buffaloes. Among it, 37.50% buffalo had mild hepatic lipidosis, 33.33% had moderate hepatic lipidosis, and 29.16% had severe hepatic lipidosis. Depth of portal vein significantly increased in hepatic lipidosis cases. However, portal vein diameter exhibited a non-significant difference in mild, moderate, and severe groups of hepatic lipidosis. Back fat thickness also revealed a non-significant difference in mild, moderate, and severe hepatic lipidosis. Above study indicate that B mode ultrasonography of the liver can be employed to differentiate various grades of hepatic lipidosis in buffaloes. Biochemical parameters like NEFA, BHBA, AST, GGT, ALP, TG, cholesterol, and glucose can be helpful to screen the hepatic lipidosis at farm level.

Correlation between liver lipidosis, body condition score variation, and hepatic analytes in dairy cows.

Liver lipidosis is a metabolic disorder mostly observed in high yielding dairy cattle, especially during the transition period. The aim of this study was to determine the correlation between hepatic lipid infiltration, biochemical indicators of liver function, and body condition score (BCS) variation in dairy cows. Fifty-one multiparous Holstein cows raised in a confined system were evaluated. Liver biopsies and blood samples were collected, and BCS was measured on days 3 and 28 postpartum. Lipid infiltration was determined by histologic examination. The plasma activity of aspartate aminotransferase, alkaline phosphatase, and gamma-glutamyl transferase and concentration of beta-hydroxybutyrate, non-esterified fatty acids, albumin, total bilirubin, and cholesterol were determined. BCS was measured using objective (camera) and subjective (visual) methods. Mild lipid infiltration was found in 3.92% of cows sampled on day 3 and 5.88% on day 28. Bilirubin was significantly higher on day 3 than on day 28 postpartum, and cholesterol was significantly higher on day 28 than on day 3 in all cows. There was no difference in biochemical analytes between cows with and without lipidosis. On day 3, mean subjective BCS was 3.10 and objective BCS was 3.16, while on day 28, these scores were 2.91 and 2.99, respectively. The calculated liver function index (LFI) was found to be a more sensitive indicator of liver function than the hepatic analytes evaluated. No correlation between BCS variation and lipid infiltration was found. Cholesterol and bilirubin levels showed the most remarkable changes during the early postpartum period. LFI is a potential indicator of postpartum liver function.

A lipidose hepática é um distúrbio metabólico principalmente observado nos bovinos de leite de alto rendimento, especialmente no período de transição. O objetivo deste estudo foi determinar a correlação entre infiltração lipídica hepática, indicadores bioquímicos de função hepática e variação da condição corporal em bovinos leiteiros. Foram avaliadas cinquenta e um vacas multíparas de raça Holandesa em confinamento. Coletaram-se biopsias de fígado e amostras de sangue, e a condição corporal (BCS) aferiu-se nos dias 3 e 28 pós-parto. A infiltração lipídica determinou-se mediante avaliação histológica. Mensuraram-se a atividade da aspartato aminotransferase, fosfatase alcalina e da gama-glutamil transferase, concentração de beta-hidroxibutirato, ácidos graxos não esterificados, albumina, bilirrubina total e colesterol. A BCS mediu-se utilizando método objetivo (câmera) e subjetivo (visualmente). Observou-se discreta infiltração lipídica em 3,92% dos bovinos amostrados no dia 3 e 5,88% no dia 28. Em todos os bovinos a bilirrubina foi significativamente mais alta no dia 3 do que no dia 28 e o colesterol foi superior no dia 28 do que no dia 3. Não houve diferença nos analitos bioquímicos dos bovinos com e sem lipidose. No dia 3, a média subjetiva da BCS foi 3,10 e a objetiva 3,16, enquanto no dia 28, obtiveram-se valores de 2,91, e 2,99 respectivamente. O índice calculado de função hepática mostrou ser um indicador mais sensível da função hepática do que os analitos avaliados individualmente. Não houve correlação entre a variação do BCS e infiltração lipídica.

Anatomopathological findings of Testudines necropsied in the Distrito Federal, Brazil / Achados anatomopatológicos de Testudines necropsiados no Distrito Federal, Brasil

In order to determine the main anatomopathological findings of Testudines necropsied in the Distrito Federal, all necropsy records performed at the "Laboratório de Patologia Veterinária" of the "Universidade de Brasília" (LPV-UnB) on Testudines during the period from January 2008 to July 2020 were reviewed. The 72 cases reviewed were grouped and classified according to species, sex, origin, season of occurrence, and diagnosis. In 69.44% of the cases the species was informed in the necropsy protocols, which included Phrynops geoffroanus (38%), Trachemys dorbigni (36%), Chelonoidis carbonaria (14%), Chelonoidis denticulata (10%) and Podocnemis expansa (2%). In 30.55% of the cases this parameter was not informed and were classified only as Testudines. In 41.66% of the cases the sex was informed, being female 22.22%, male 19.44%, and 58.33% were not informed. Of these animals 79.16% were from environmental agencies and 20.84% from zoos and/or guardians. In 70.83% of the animals analyzed they were directly related to the autumn and winter seasons, with June being the most frequent month (29.17%). The conclusive diagnosis was possible in 68.05% of the cases. The category of disorders caused by injurious agents (48.97%) was the most prevalent, followed by inflammatory disorders (32.65%) and nutritional and metabolic disorders (28.57%). The main diagnoses were carapace and/or plastron fracture with 30.61%, hepatic steatosis (20.40%) and pneumonia (10.22%). Most cases of carapace or plastron fracture and hepatic steatosis occurred in animals from environmental agencies.

Com o objetivo de determinar os principais achados anatomopatológicos de Testudines necropsiados no Distrito Federal, foram revisadas todas as fichas de necropsia realizadas no Laboratório de Patologia Veterinária da Universidade de Brasília (LPV-UnB) em Testudines, durante o período de janeiro de 2008 a julho de 2020. Os 72 casos revisados foram agrupados e classificados quanto à espécie, sexo, procedência, estação do ano de ocorrência e diagnóstico. Em 69,44% dos casos havia a espécie informada nos protocolos de necropsia, que incluíam Phrynops geoffroanus (38%), Trachemys dorbigni (36%), Chelonoidis carbonaria (14%), Chelonoidis denticulata (10%) e Podocnemis expansa (2%). Em 30,55% dos casos não tiveram esse parâmetro informado e foram classificados apenas como Testudines. Em 41,66% casos foi informado o sexo, sendo fêmea 22,22%, macho 19,44% e não informados 58,33%. Destes animais 79,16% eram de órgão ambiental e 20,84% de zoológicos e ou tutores. Em 70,83% dos animais analisados tiveram direta relação com as estações de outono e inverno, sendo o mês de junho o mais frequente (29,17%). O diagnóstico conclusivo foi possível em 68,05% dos casos. A categoria de distúrbios causados por agentes lesivos (48,97%) foi a mais prevalente, seguido por distúrbios inflamatórios (32,65%) e dos distúrbios nutricionais e metabólicos (28,57%). Os principais diagnósticos foram fratura de carapaça e ou plastrão com 30,61%, esteatose hepática (20,40%) e pneumonia (10,22%). A maior parte dos casos de fratura de carapaça ou plastrão e de esteatose hepática ocorreram em animais provenientes de órgão ambiental.

Anatomopathological findings of Testudines necropsied in the Distrito Federal, Brazil

ABSTRACT: In order to determine the main anatomopathological findings of Testudines necropsied in the Distrito Federal, all necropsy records performed at the Laboratório de Patologia Veterinária of the Universidade de Brasília (LPV-UnB) on Testudines during the period from January 2008 to July 2020 were reviewed. The 72 cases reviewed were grouped and classified according to species, sex, origin, season of occurrence, and diagnosis. In 69.44% of the cases the species was informed in the necropsy protocols, which included Phrynops geoffroanus (38%), Trachemys dorbigni (36%), Chelonoidis carbonaria (14%), Chelonoidis denticulata (10%) and Podocnemis expansa (2%). In 30.55% of the cases this parameter was not informed and were classified only as Testudines. In 41.66% of the cases the sex was informed, being female 22.22%, male 19.44%, and 58.33% were not informed. Of these animals 79.16% were from environmental agencies and 20.84% from zoos and/or guardians. In 70.83% of the animals analyzed they were directly related to the autumn and winter seasons, with June being the most frequent month (29.17%). The conclusive diagnosis was possible in 68.05% of the cases. The category of disorders caused by injurious agents (48.97%) was the most prevalent, followed by inflammatory disorders (32.65%) and nutritional and metabolic disorders (28.57%). The main diagnoses were carapace and/or plastron fracture with 30.61%, hepatic steatosis (20.40%) and pneumonia (10.22%). Most cases of carapace or plastron fracture and hepatic steatosis occurred in animals from environmental agencies.

RESUMO: Com o objetivo de determinar os principais achados anatomopatológicos de Testudines necropsiados no Distrito Federal, foram revisadas todas as fichas de necropsia realizadas no Laboratório de Patologia Veterinária da Universidade de Brasília (LPV-UnB) em Testudines, durante o período de janeiro de 2008 a julho de 2020. Os 72 casos revisados foram agrupados e classificados quanto à espécie, sexo, procedência, estação do ano de ocorrência e diagnóstico. Em 69,44% dos casos havia a espécie informada nos protocolos de necropsia, que incluíam Phrynops geoffroanus (38%), Trachemys dorbigni (36%), Chelonoidis carbonaria (14%), Chelonoidis denticulata (10%) e Podocnemis expansa (2%). Em 30,55% dos casos não tiveram esse parâmetro informado e foram classificados apenas como Testudines. Em 41,66% casos foi informado o sexo, sendo fêmea 22,22%, macho 19,44% e não informados 58,33%. Destes animais 79,16% eram de órgão ambiental e 20,84% de zoológicos e ou tutores. Em 70,83% dos animais analisados tiveram direta relação com as estações de outono e inverno, sendo o mês de junho o mais frequente (29,17%). O diagnóstico conclusivo foi possível em 68,05% dos casos. A categoria de distúrbios causados por agentes lesivos (48,97%) foi a mais prevalente, seguido por distúrbios inflamatórios (32,65%) e dos distúrbios nutricionais e metabólicos (28,57%). Os principais diagnósticos foram fratura de carapaça e ou plastrão com 30,61%, esteatose hepática (20,40%) e pneumonia (10,22%). A maior parte dos casos de fratura de carapaça ou plastrão e de esteatose hepática ocorreram em animais provenientes de órgão ambiental.

Anatomopathological findings of Testudines necropsied in the Distrito Federal, Brazil / Achados anatomopatológicos de Testudines necropsiados no Distrito Federal, Brasil

In order to determine the main anatomopathological findings of Testudines necropsied in the Distrito Federal, all necropsy records performed at the "Laboratório de Patologia Veterinária" of the "Universidade de Brasília" (LPV-UnB) on Testudines during the period from January 2008 to July 2020 were reviewed. The 72 cases reviewed were grouped and classified according to species, sex, origin, season of occurrence, and diagnosis. In 69.44% of the cases the species was informed in the necropsy protocols, which included Phrynops geoffroanus (38%), Trachemys dorbigni (36%), Chelonoidis carbonaria (14%), Chelonoidis denticulata (10%) and Podocnemis expansa (2%). In 30.55% of the cases this parameter was not informed and were classified only as Testudines. In 41.66% of the cases the sex was informed, being female 22.22%, male 19.44%, and 58.33% were not informed. Of these animals 79.16% were from environmental agencies and 20.84% from zoos and/or guardians. In 70.83% of the animals analyzed they were directly related to the autumn and winter seasons, with June being the most frequent month (29.17%). The conclusive diagnosis was possible in 68.05% of the cases. The category of disorders caused by injurious agents (48.97%) was the most prevalent, followed by inflammatory disorders (32.65%) and nutritional and metabolic disorders (28.57%). The main diagnoses were carapace and/or plastron fracture with 30.61%, hepatic steatosis (20.40%) and pneumonia (10.22%). Most cases of carapace or plastron fracture and hepatic steatosis occurred in animals from environmental agencies.(AU)

Com o objetivo de determinar os principais achados anatomopatológicos de Testudines necropsiados no Distrito Federal, foram revisadas todas as fichas de necropsia realizadas no Laboratório de Patologia Veterinária da Universidade de Brasília (LPV-UnB) em Testudines, durante o período de janeiro de 2008 a julho de 2020. Os 72 casos revisados foram agrupados e classificados quanto à espécie, sexo, procedência, estação do ano de ocorrência e diagnóstico. Em 69,44% dos casos havia a espécie informada nos protocolos de necropsia, que incluíam Phrynops geoffroanus (38%), Trachemys dorbigni (36%), Chelonoidis carbonaria (14%), Chelonoidis denticulata (10%) e Podocnemis expansa (2%). Em 30,55% dos casos não tiveram esse parâmetro informado e foram classificados apenas como Testudines. Em 41,66% casos foi informado o sexo, sendo fêmea 22,22%, macho 19,44% e não informados 58,33%. Destes animais 79,16% eram de órgão ambiental e 20,84% de zoológicos e ou tutores. Em 70,83% dos animais analisados tiveram direta relação com as estações de outono e inverno, sendo o mês de junho o mais frequente (29,17%). O diagnóstico conclusivo foi possível em 68,05% dos casos. A categoria de distúrbios causados por agentes lesivos (48,97%) foi a mais prevalente, seguido por distúrbios inflamatórios (32,65%) e dos distúrbios nutricionais e metabólicos (28,57%). Os principais diagnósticos foram fratura de carapaça e ou plastrão com 30,61%, esteatose hepática (20,40%) e pneumonia (10,22%). A maior parte dos casos de fratura de carapaça ou plastrão e de esteatose hepática ocorreram em animais provenientes de órgão ambiental.(AU)

Hepatic effects of rumen-protected branched-chain amino acids with or without propylene glycol supplementation in dairy cows during early lactation.

Essential amino acids (EAA) are critical for multiple physiological processes. Branched-chain amino acid (BCAA) supplementation provides energy substrates, promotes protein synthesis, and stimulates insulin secretion in rodents and humans. Most dairy cows face a protein and energy deficit during the first weeks postpartum and utilize body reserves to counteract this shortage. The objective was to evaluate the effect of rumen-protected BCAA (RP-BCAA; 375 g of 27% l-leucine, 85 g of 48% l-isoleucine, and 91 g of 67% l-valine) with or without oral propylene glycol (PG) administration on markers of liver health status, concentrations of nonesterified fatty acids (NEFA) and ß-hydroxybutyrate (BHB) in plasma, and liver triglycerides (TG) during the early postpartum period in dairy cows. Multiparous Holstein cows were enrolled in blocks of 3 and randomly assigned to either the control group or 1 of the 2 treatments from calving until 35 d postpartum. The control group (n = 16) received 200 g of dry molasses per cow/d; the RP-BCAA group (n = 14) received RP-BCAA mixed with 200 g of dry molasses per cow/d; the RP-BCAA plus PG (RP-BCAAPG) group (n = 16) received RP-BCAA mixed with 200 g of dry molasses per cow/d, plus 300 mL of PG, once daily from calving until 7 d in milk (DIM). The RP-BCAA and RP-BCAAGP groups, on average (± standard deviation), were predicted to receive a greater supply of metabolizable protein in the form of l-Leu 27.4 ± 3.5 g/d, l-Ile 15.2 ± 1.8 g/d, and l-Val 24.2 ± 2.4 g/d compared with the control cows. Liver biopsies were collected at d 9 ± 4 prepartum and at 5 ± 1 and 21 ± 1 DIM. Blood was sampled 3 times per week from calving until 21 DIM. Milk yield, dry matter intake, NEFA, BHB, EAA blood concentration, serum chemistry, insulin, glucagon, and liver TG and protein abundance of total and phosphorylated branched-chain ketoacid dehydrogenase E1α (p-BCKDH-E1α) were analyzed using repeated measures ANOVA. Cows in the RP-BCAA and RP-BCAAPG groups had lower liver TG and lower activities of aspartate aminotransferase and glutamate dehydrogenase during the first 21 DIM, compared with control. All cows, regardless of treatment, showed an upregulation of p-BCKDH-E1α at d 5 postpartum, compared with levels at 21 d postpartum. Insulin, Met, and Glu blood concentration were greater in RP-BCAA and RP-BCAAPG compared with control during the first 35 DIM. Therefore, the use of RP-BCAA in combination with PG might be a feasible option to reduce hepatic lipidosis in dairy cows during early lactation.

Hepatic lipidosis: Liver characteristics and acute phase proteins in affected turkeys.

The hepatic lipidosis (HL) in fattening turkeys is a disease has been known for a long time, but the cause and pathogenesis is still not clarified. A recent study reported unexplained high levels of iron in liver tissue of fattening turkeys suffering from HL. In this study, the iron status, possible infectious or inflammatory influences in form of an acute phase reaction and the analysis of fatty acid pattern in liver tissue of turkeys affected by HL were examined. Three cases of HL on three different fattening turkey farms were investigated during the outbreak of the disease. Clinically affected and non-affected animals were subjected to a pathological examination, where the diagnosis HL or non-affected was made. In total, 70 birds were examined (40 with HL, 30 without HL) and blood and liver samples were taken. Additionally, samples from 15 slaughtered birds were taken as a further control group. In liver tissue, the iron content and the content of long-chain fatty acids were determined; in blood samples, ferritin and transferrin were measured. The iron content in liver tissue was more than three times higher for animals with HL than among non-affected animals and the control group. The transferrin levels were lowest for animals with HL, highest in the control group and in between for non-affected animals. The fatty acid pattern in liver tissue of affected animals indicated a shift from polyunsaturated fatty acids to saturated fatty acids and monounsaturated fatty acids compared to the control group and the non-affected animals. Overall, the non-affected animals of a flock affected by HL were similar to the healthy animals of the abattoir. The low acute phase protein levels for animals with HL together with high iron contents could indicate a previous malnutrition/starvation period and/or severe liver damage for those animals suffering from HL.

Hepatogenous Photosensitivity Outbreak after Coccidiosis in Grazing Holstein Steers.

Clinical signs of photosensitivity in cattle can occur sporadically and unpredictably. It is believed that cases of photosensitivity may be underreported, causing inaccurate and inflated reports of mortality. Additionally, because secondary photosensitization in grazing cattle occurs with liver damage or dysfunction, photosensitivity can have many potential or associated causes. This case links a previous occurrence of coccidiosis to an outbreak of photosensitivity in grazing Holstein steers. Grazing management staff first observed clinical signs of photosensitivity 17 days after an outbreak of coccidiosis and subsequent turnout to spring pastures. Clinical signs were observed in 25% of the population. The severity of photosensitivity was variable and ranged from blistered skin on the muzzle to sloughing of unpigmented epidermis and thinly haired regions. Severely affected cattle were removed from pasture, housed under shade, monitored for infection, and recovered without treatment. Mild cases remained on pasture and recovered without treatment. Photosensitivity did not reoccur in the cattle that remained on pasture or in mildly affected cattle returned to pasture. Photosensitivity did not appear to be associated with pasture weeds, a specific forage species, or variable or extreme weather conditions that could have resulted in mycotoxin production. The occurrence appears to have been a result of a previous and concurrent coccidiosis outbreak that caused secondary photosensitization through hepatic lipidosis caused by anorexia and dehydration associated with the severe coccidiosis. Although clinical signs appeared suddenly, cattle recovered quickly and without treatment.

Physical exercise prepartum to support metabolic adaptation in the transition period of dairy cattle: A proof of concept.

In dairy cattle, the hormonal changes around calving induce large metabolic changes to support milk production. Mobilization of adipose reserves is one of the changes involved, imposing a metabolic load on the liver. We hypothesized that the risk for excessive lipolysis and hepatic lipidosis postpartum can be reduced by starting fat mobilization and processing during the prepartum period by physical exercise, especially in cows with a high body condition score (BCS). As a proof of concept, 32 pregnant Holstein-Friesian dairy cows were selected for a 2 × 2 experimental design. Sixteen cows had a BCS < 3.25 (group LOW) and 16 cows a BCS ≥ 3.25 (group HIGH). Cows within each group were randomly allocated to one of two treatments: group STEP was walked twice daily for 45 min during the dry period while group CON remained indoors. Treatment was stopped at calving and cows were monitored until 6 weeks after calving. Liver biopsies were taken in a subset of 16 cows to determine liver triglyceride (TG) concentration. We found that calculated energy balance was more negative for group STEP prepartum, resulting in higher plasma non-esterified fatty acids and ß-hydroxybutyrate concentrations. During the first 6 weeks postpartum, neither dry matter intake nor milk yield was affected by exercise. As expected, the cows in group HIGH had increased liver TG concentrations postpartum relative to group LOW with increased plasma non-esterified fatty acids directly after calving. Exercise during the dry period mitigated postpartal liver TG accumulation, but this did not seem to be related to increased plasma lipoprotein transport. We conclude that substantial physical activity prepartum can induce lipolysis and lipid utilization, thereby starting an early adaptation to lactation. This may be instrumental to reduce the risk for excessive liver TG accumulation postpartum, especially in cows with a high BCS at dry-off.

Serum vitamin D status in sick cats with and without cholestatic liver disease.

OBJECTIVES: Vitamin D deficiency accompanies chronic cholestatic liver disease (CLD) in humans. The vitamin D status of cats with CLD is unknown. The objectives of this study were to describe serum vitamin D concentrations in cats with CLD and to determine if they correlated with indices of liver disease severity. METHODS: Thirty-six cats with CLD, defined by increases in serum bilirubin and serum alanine aminotransferase, and 23 sick cats with non-hepatobiliary diseases were prospectively enrolled. Serum 25-hydroxyvitamin D (25[OH]D), parathyroid hormone (PTH) and ionized calcium were measured. Signalment, clinical signs, comorbidities, diet history, serum bilirubin, liver enzyme activity, albumin, phosphorus, white blood cell count, prothrombin time and final hepatic cytologic/histopathologic diagnosis, when available, were recorded. RESULTS: Median serum 25(OH)D levels were similar in cats with CLD (89.5 nmol/l; range 21-112 nmol/l) and sick cats (89.0 nmol/l; range 49-115 nmol/l). Overall 12/36 (33%) cats with CLD and 4/23 (17%) sick cats had 25(OH)D levels below the lower limit of the reference interval (<65 nmol/l). Median PTH concentrations in cats with CLD were significantly higher (0.95 pmol/l; range 0-11.3 pmol/l) than in sick cats (0.70 pmol/l; range 0.5-6 pmol/l). In cats with CLD, 6/36 (17%) had high PTH levels in contrast to only 1/23 (4%) sick cats. In cats with CLD, 25(OH)D concentrations did not correlate with serum bilirubin, albumin or serum liver enzymes but were moderately negatively correlated with white blood cell count (r = - 0.402, P = 0.013). Cats with hepatic lipidosis had the highest prevalence of 25(OH)D concentrations that fell below the reference interval. CONCLUSIONS AND RELEVANCE: Many cats with CLD have serum 25(OH)D concentrations below the lower limit of the reference interval. Further study is warranted to determine the clinical relevance and whether supplementation would provide benefits.

Gene expressions of de novo hepatic lipogenesis in feline hepatic lipidosis.

OBJECTIVES: The aim of this study was to evaluate if de novo hepatic lipid synthesis contributes to fatty acid overload in the liver of cats with feline hepatic lipidosis (FHL). METHODS: Lipogenic gene expression of peroxisome proliferator-activated receptor-alpha (PPAR-α), peroxisome proliferator-activated receptor-gamma (PPAR-γ), fatty acid synthase (FASN) and sterol regulatory element-binding factor (SREBF1) were evaluated using quantitative RT-PCR in liver tissue of six cats with FHL and compared with the liver tissue of eight healthy cats. RESULTS: In liver tissue, PPAR-α, PPAR-γ and FASN mRNA expression levels were not significantly different (P >0.12, P >0.89 and P >0.5, respectively) in the FHL group compared with the control group. SREBF1 gene expression was downregulated around 10-fold in the FHL group vs the control group (P = 0.039). CONCLUSIONS AND RELEVANCE: The downregulation of SREBF1 in the liver tissue of cats with FHL does not support the hypothesis that de novo lipogenesis in the liver is an important pathway of fatty acid accumulation in FHL.

Hepatic CT attenuation differs in three species of freshwater turtles and hepatic Hounsfield units increase with folliculogenesis in wild Blanding's turtles (Emydoidea blandingii).

Freshwater turtle species are suffering from anthropocentric-caused population declines, making preservation of professionally managed populations increasingly important. Turtles under professional care have an increased risk to develop hepatic lipidosis, potentially resulting in early death. Computed tomography can provide an antemortem screening for increased fatty liver composition. A goal of this prospective analytical cross-sectional study was to assess the hepatic attenuation measured as Hounsfield units (HU) in a wild population of a freshwater turtle species, and then compare hepatic HU to freshwater turtles under professional care. Ninety-five wild Blanding's turtles (BT; Emydoidea blandingii) as well as 10 Vietnamese Pond turtles (VPT; Mauremys annamensis) and six Northern Snake-Necked turtles (NSNT; Chelodina oblonga) under professional care underwent CT with quantification of hepatic HU. Hepatic HU were correlated to serum chemistry findings and the presence of follicles was recorded in BT. The mean (±SD) hepatic attenuation for 95 wild BT was 97.5 HU ±9.6. There were significant differences in hepatic attenuation among VPT, NSNT, and BT, with median HU values (range) of 5.39 HU (-6.45 to 61.50), 71.74 HU (59.44-94.49), and 95.43 HU (74.41-124.56), respectively. Aspartate aminotransferase (AST) values had a weak negative correlation to HU with a coefficient of -0.85 (P < .001). The hepatic attenuation was significantly higher for individuals undergoing folliculogenesis (P = .007). The decreased HU values were significantly negatively correlated with AST values. Findings supported the use of CT as an aid for guiding future management practices in freshwater turtles.

Retrospective study of the relative frequency of feline hepatobiliary disease in New Zealand based on 10 years of hepatic biopsy samples.

AIMS: To retrospectively determine the relative frequency of feline hepatobiliary diseases from biopsy specimens submitted to a single laboratory across a 10-year period and to establish whether age, sex or breed associations exist. METHODS: Histopathological data from 154 liver biopsies of New Zealand cats sampled between 2008 and 2018 were analysed. The samples were allocated to primary, secondary and tertiary disease categories using criteria established by the World Small Animal Veterinary Association. Breed associations were derived using ORs and 95% CIs. Gender and age associations were also evaluated. RESULTS: The most frequently diagnosed hepatobiliary diseases were lymphocytic cholangitis (20 per cent), hepatitis (16.9 per cent), reversible hepatocellular injury (16.4 per cent), neutrophilic cholangitis (9.7 per cent), haematopoietic neoplasia (9.7 per cent), hepatocellular neoplasia (5.6 per cent) and cholangiocellular neoplasia (4.1 per cent). Burmese cats were found to be at significantly increased risk of both biliary and parenchymal diseases and Birman cats to be at significantly increased risk of parenchymal disease. Domestic longhair cats were at significantly increased risk of hepatobiliary neoplasia. Birman cats were at significantly increased risk of hepatitis while domestic shorthair cats were at significantly decreased risk of neutrophilic cholangitis, reversible hepatocellular injury and hepatitis. CONCLUSIONS: This study is the first retrospective examination of the relative frequency of hepatobiliary disease in biopsy specimens from New Zealand cats. Some breeds were associated with specific histopathology.

Amino acid pattern in the liver and blood of fattening turkeys suffering from hepatic lipidosis.

Hepatic lipidosis (HL) is a well-known disease in fattening and in parent turkey flocks. Among others, dietary effects like (a lack of) essential amino acids (AA) as lipotrophic factors (e.g., methionine) have been considered as potentially predispositing for HL. Several studies have reported abnormal AA profiles in hepatic diseases of humans and other livestock. The ratio of branched-chain amino acids (BCAA) to aromatic amino acids (AAA) in plasma is used to predict hepatic cirrhosis. In this study, the state of supply of AA was investigated by comparing non-affected (NA) animals and those affected by HL. The AA pattern in the liver and blood can provide potential indications of pathogenesis of HL. In cooperation with German poultry veterinarians, 3 cases of HL on 3 different fattening turkey farms were visited (13/14 wk old, "B.U.T. Big 6" and "TP7"). Overall, 73 birds were examined, of which 42 birds suffered from HL and 31 were not affected. Feeding samples of the respective actual feed were taken and analyzed. The selection of animals was carried out (NA randomly) by clinical signs such as apathy and dyspnea and the diagnosis was made at necropsy, which could be confirmed by crude fat content in liver tissue (HL: 309, NA: 155). In liver tissue, the CP and AA contents were lower among animals with HL than among NA (P < 0.05). In blood samples, the sum of AA, ammonia, and urea was more than 3 times higher among animals with HL (431 mg/dL serum) than among NA (114 mg/dL serum; P < 0.01). The ratio of BCAA to AAA was also significantly different between the groups (HL: 0.85, NA: 1.42; P < 0.05). In the case of HL, entire herds were not affected and the "non-affected" ones were comparable with healthy slaughtered animals. There seems to be a clear change in protein and AA metabolism of HL animals, which could lead to an optimization in feeding practice in repeated cases of HL.

Lipid-Related Lesions in Quaker Parrots ( Myiopsitta monachus).

The Quaker parrot has been used as a psittacine model to study clinical lipidology and lipid-related disorders. However, while Quaker parrots appear to be anecdotally susceptible to a variety of spontaneous dyslipidemic disorders and lesions caused by excess lipid accumulation, epidemiologic data are lacking. A multicenter retrospective study on 652 pathology submissions (411 necropsies and 243 biopsies) from Quaker parrots was performed by recording the final pathological diagnoses, age, and sex for each bird. The prevalence of lesions associated with lipid metabolism, such as hepatic lipidosis, atherosclerosis, xanthomas, adipose tumors, coelomic steatitis/steatonecrosis, endogenous lipid pneumonia, and acute pancreatic necrosis/pancreatitis, was reported. Multiple logistic regression models were used to characterize the effects of sex and age on these lesions, and the prevalence of hepatic lipidosis and atherosclerosis was compared to those in a random sample of control psittacine birds. The raw prevalence of atherosclerosis and hepatic lipidosis was 5.6% (95% confidence interval [CI], 3.4%-7.8%) and 21.2% (95% CI, 17.2%-25.1%), respectively. While the prevalence of atherosclerosis was similar to other psittacine species, hepatic lipidosis was more common in Quaker parrots. Quaker parrots also showed a unique susceptibility to acute pancreatic necrosis with a prevalence of 12.9% (95% CI, 9.7%-16.1%). Male parrots were found to be more susceptible than females to lipid accumulation lesions ( P = .0024), including atherosclerosis ( P = .018) and hepatic lipidosis ( P < .001). This retrospective study confirms the high susceptibility of Quaker parrots to lipid-related disorders and presents epidemiological data that may be useful to avian clinicians, pathologists, and researchers using Quaker parrots.