Liver Tissue Concentrations Research Articles

Protective Effects of Fucoxanthin against Alcoholic Liver Injury by Activation of Nrf2-Mediated Antioxidant Defense and Inhibition of TLR4-Mediated Inflammation.

Fucoxanthin (Fx) is a natural extract from marine seaweed that has strong antioxidant activity and a variety of other bioactive effects. This study elucidated the protective mechanism of Fx on alcoholic liver injury. Administration of Fx was associated with lower pathological effects in liver tissue and lower serum marker concentrations for liver damage induced by alcohol. Fx also alleviated oxidative stress, and lowered the level of oxides and inflammation in liver tissue. Results indicate that Fx attenuated alcohol-induced oxidative lesions and inflammatory responses by activating the nuclear factor erythrocyte-2-related factor 2 (Nrf2)-mediated signaling pathway and down-regulating the expression of the toll-like receptor 4 (TLR4)-mediated nuclear factor-kappa B (NF-κB) signaling pathway, respectively. Our findings suggest that Fx can be developed as a potential nutraceutical for preventing alcohol-induced liver injury in the future.

Bile acid-polymer-probucol microparticles: protective effect on pancreatic β-cells and decrease in type 1 diabetes development in a murine model

Studies in our laboratory have shown potential applications of the anti-atherosclerotic drug probucol (PB) in diabetes due to anti-inflammatory and β-cell protective effects. The anti-inflammatory effects were optimized by incorporation of the anti-inflammatory bile acid, ursodeoxycholic acid (UDCA). This study aimed to test PB absorption, tissue accumulation profiles, effects on inflammation and type 1 diabetes prevention when combined with UDCA. Balb/c mice were divided into three equal groups and gavaged daily PB powder, PB microcapsules or PB-UDCA microcapsules for one week, at a constant dose. Mice were injected with a single dose of intraperitoneal/subcutaneous alloxan to induce type-1 diabetes and once diabetes was confirmed, treatments were continued for 3 days. Mice were euthanized and blood and tissues collected for analysis of PB and cytokine levels. The PB-UDCA group showed the highest PB concentrations in blood, gut, liver, spleen, brain, and white adipose tissues, with no significant increase in pancreas, heart, skeletal muscles, kidneys, urine or feces. Interferon gamma in plasma was significantly reduced by PB-UDCA suggesting potent anti-inflammatory effects. Blood glucose levels remained similar after treatments, while survival was highest among the PB-UDCA group. Our findings suggest that PB-UDCA resulted in best PB blood and tissue absorption and reduced inflammation.

Characteristics of Changes in Inflammatory Cytokines as a Function of Hepatic Ischemia-Reperfusion Injury Stage in Mice.

Liver ischemia-reperfusion injury (IRI) can severely compromise the prognosis of patients receiving liver surgery. While inflammation contributes to the damage resulting from IRI, only a limited number of inflammation biomarkers have been identified as being associated with the different stages of hepatic IRI. As an approach to identify some of these inflammatory cytokines and the molecular mechanisms involved within different stages of hepatic IRI, we used an advanced antibody array assay to detect multiple proteins. With this technology, we observed specific differences in the content of inflammatory cytokines between ischemic and sham controls, as well as a function of the different reperfusion stages in a hepatic IRI mouse model. For example, while liver tissue content of IL-12p40/p70 was significantly increased in the ischemic stage, it was significantly decreased in the reperfusion stage as compared with that of the sham group. For other inflammatory cytokines, no changes were obtained between the ischemic and reperfusion stages with levels of IL-17, Eotaxin-2, Eotaxin, and sTNF RII all being consistently increased, while those of TIMP-1, TIMP-2, BLC, and MCSF consistently decreased as compared with that of the sham group at all reperfusion stages examined. Results from protein function annotation Gene Ontology and the KEGG pathway revealed that inflammatory cytokines are enriched in a network associated with activation of inflammatory response signaling pathways such as TLR, TNF, and IL-17 when comparing responses of the IR versus sham groups. The identification of cytokines along with their roles at specific stages of IRI may reveal important new biological markers for the diagnosis and prognosis of hepatic IRI.

Indole-3-Acetic Acid Alleviates Nonalcoholic Fatty Liver Disease in Mice via Attenuation of Hepatic Lipogenesis, and Oxidative and Inflammatory Stress.

Recent evidences have linked indole-3-acetic acid (IAA), a gut microbiota-derived metabolite from dietary tryptophan, with the resistance to liver diseases. However, data supporting IAA-mediated protection against nonalcoholic fatty liver disease (NAFLD) from an in vivo study is lacking. In this study, we assessed the role of IAA in attenuating high-fat diet (HFD)-induced NAFLD in male C57BL/6 mice. Administration of IAA (50 mg/kg body weight) by intraperitoneal injection was found to alleviate HFD-induced elevation in fasting blood glucose and homeostasis model assessment of insulin resistance (HOMA-IR) index as well as plasma total cholesterol, low-density lipoprotein cholesterol (LDL-C), and glutamic-pyruvic transaminase (GPT) activity. Histological examination further presented the protective effect of IAA on liver damage induced by HFD feeding. HFD-induced an increase in liver total triglycerides and cholesterol, together with the upregulation of genes related to lipogenesis including sterol regulatory element binding-protein 1 (Srebf1), steraroyl coenzyme decarboxylase 1 (Scd1), peroxisome proliferator-activated receptor gamma (PPARγ), acetyl-CoA carboxylase 1 (Acaca), and glycerol-3-phosphate acyltransferase, mitochondrial (Gpam), which were mitigated by IAA treatment. The results of reactive oxygen species (ROS) and malonaldehyde (MDA) level along with superoxide dismutase (SOD) activity and glutathione (GSH) content in liver tissue evidenced the protection of IAA against HFD-induced oxidative stress. Additionally, IAA attenuated the inflammatory response of liver in mice exposed to HFD as shown by the reduction in the F4/80-positive macrophage infiltration and the expression of monocyte chemoattractant protein-1 (MCP-1) and tumor necrosis factor-α (TNF-α). In conclusion, our findings uncover that IAA alleviates HFD-induced hepatotoxicity in mice, which proves to be associated with the amelioration in insulin resistance, lipid metabolism, and oxidative and inflammatory stress.

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.

Panax ginseng is superior to vitamin E as a hepatoprotector against cyclophosphamide-induced liver damage

Background and aimsCyclophosphamide (CPh) is a frequently used drug, in human and animals for its immunosuppressive and anticancer potential. However, it is metabolized by the liver yielding damaging toxicants (to the liver itself and other non-target vital organs) via oxidative stress, apoptosis induction and finally necrosis. Since there is no escaping of using such harmful medications, we focused on alleviating its side-effects. Panax ginseng Meyer is a potent candidate, and we still lack adequate information on its hepatoprotective role against cyclophosphamide-induced liver-damage. MethodsHere, we used P. ginseng (Korean Red Ginseng) compared to vitamin-E (natural antioxidant) in combating CPh-induced liver damage. Forty-eight albino rats were divided into 6 groups, Control, Ginseng, Vitamin E, Cyclophosphamide (CPh), CPh + Ginseng or CPh + Vitamin-E. Blood samples were taken for biochemical analyses and liver samples were collected for histopathology, oxidative stress evaluation, and gene expression analyses. ResultsIn CPh group, typical CPh-liver damage was evident (higher levels of AST, ALT, ALP; lower albumin and total proteins levels; lower liver tissue concentrations of SOD, GPX and CAT and higher MDA; injured liver histopathological picture; and finally increased TNF-α, IL-1β and Caspase3 and decreased BCL-2 genes expression). All these were abolished with either P. ginseng or vitamin-E administration. However, P. ginseng was overall superior to vitamin-E, especially in restoring blood biochemical findings and damaged histopathological picture. ConclusionsTherefore, P. ginseng is a potent hepatoprotector (vitamin-E to a lesser extent) and should be considered where liver damage is expected secondary to damaging medications; as cyclophosphamide.

The Protective Effect of Aesculus hippocastanum (Venoplant®) Against Concanavalin A-Induced Liver Injury

Aim: The present study was performed to investigate the effect of Aesculus hippocastanum (AH; Venoplant®) on concanavalin A (ConA)-induced acute liver injury and explore the mechanism in mice. Methods: ConA (20 mg/kg) was administered via tail vein injection to induce hepatic damage. The groups of AH (Venoplant®) were given at 65.8, 131.6, and 263.2 mg/kg by oral gavages for 20 days. The serum levels of aspartate transaminase (AST), alanine aminotransferase (ALT), total protein (TP), and albumin (Alb) were determined by automatic biochemical analyzer, and the Alb/globulin (A/G) ratio was calculated. Tumor necrosis factor-α (TNF-α) and IFN-γ levels were assayed by enzyme-linked immunosorbent assay. The liver tissue was attained by hematoxylin and eosin, and the histopathological changes were calculated. The cell apoptosis was assayed by terminal dUTP nick-end labeling. The malondialdehyde (MDA), superoxide dismutase (SOD), and glutathione (GSH) content of liver tissue were assayed by related kits. The activity of caspase-3 was detected by spectrophotometry. The expressions of cytochrome c, Bax, Bcl-2, c-Jun N-terminal kinase (JNK), and p-JNK were detected by western blot. Results: The results showed that the levels of ALT, AST, IFN-γ, and TNF-α in AH (Venoplant®) groups were significantly lower than those in ConA-injured group, while the levels of TP, Alb, and A/G were significantly higher. The SOD and GSH levels were significantly increased, and the MDA level was decreased; liver histopathology was changed consistently with the serological indicators, AH (Venoplant®) treatment significantly reduced the pathological damage and cell apoptosis; while in AH (Venoplant®) group, the expressions of cytochrome c, caspase-3, Bax/Bcl-2 ratio, and p-JNK were significantly decreased. Conclusion: AH (Venoplant®) could significantly protect the ConA-induced acute liver injury in mice via inhibition of reactive oxygen species and JNK pathway.

Hepatic betaine-homocysteine methyltransferase and methionine synthase activity and intermediates of the methionine cycle are altered by choline supply during negative energy balance in Holstein cows

Although choline requirements are unknown, enhanced postruminal supply may decrease liver triacylglycerol (TAG) storage and increase flux through the methionine cycle, helping cows during a negative energy balance (NEB). The objective was to investigate effects of postruminal choline supply during NEB on hepatic activity of betaine-homocysteine methyltransferase (BHMT), methionine synthase (MTR), methionine adenosyltransferase, transcription of enzymes, and metabolite concentrations in the methionine cycle. Ten primiparous rumen-cannulated Holstein cows (158 ± 24 d postpartum) were used in a replicated 5 × 5 Latin square design with 4-d treatment periods and 10 d of recovery (14 d/period). Treatments were unrestricted intake with abomasal infusion of water (A0), restricted intake (R; 60% of net energy for lactation requirements to induce NEB) with abomasal infusion of water (R0) or R plus abomasal infusion of 6.25, 12.5, or 25 g/d of choline ion. Liver tissue was collected on d 5 after the infusions ended, blood on d 1 to 5, and milk on d 1 to 4. Statistical contrasts were A0 versus R0 (CONT1) and tests of linear (L), quadratic (Q), and cubic (C) effects of choline dose. Plasma choline increased with R (CONT1) and choline (L). Although R decreased milk yield (CONT1), choline increased milk yield and liver phosphatidylcholine (PC), but decreased TAG (L). No differences were observed in plasma PC or very-low-density lipoprotein concentrations with R or choline. Activity and mRNA abundance of BHMT were greater with R (CONT1) and increased with choline (L). Although activity of MTR was lower with R (CONT1), it tended to increase with choline (L). No effect of R was detected for activity of methionine adenosyltransferase, but it changed cubically across dose of choline. Those responses were associated with linear increases in the concentrations of liver tissue (+13%) and plasma methionine concentrations. The mRNA abundance of CPT1A, SLC22A5, APOA5, and APOB, genes associated with fatty acid oxidation and lipoprotein metabolism, was upregulated by choline (Q). Overall, enhanced supply of choline during NEB increases hepatic activity of BHMT and MTR to regenerate methionine and PC, partly to help clear TAG. The relevance of these effects during the periparturient period merits further research.

Hepatoprotective Activity of Caspian Saffron (Crocus caspius Fisch and Mey) Flowers against CCl4- Induced Acute Liver Injury in Mice

Background and objectivesFree radicals and other oxidants have important roles in liver cell toxicity. Some natural products are potent scavengers of oxidative agents. Crocus caspius is an endemic species of Caspian forest. Flowers of Crocus genus contain phenolic compound and carotenoid as antioxidant agent. This study investigated the antioxidant and hepatoprotective activity of Crocus caspius Fisch and Mey flowers (Caspian saffron) hydro-alcoholic extract (CCFE) against acute oxidative hepatotoxicity induced by CCl4 in mice.MethodsThe antioxidant activity of the extract was evaluated by 2, 2-diphenyl-1-picryl hydrazyl (DPPH) radical scavenging assay. Crocus caspius flowers were investigated for presence of certain phytochemicals and total phenol and flavonoid contents were determined. For evaluation of hepatoprotective activity, the BALB/c mice were pre-treated with 50, 100, 200, and 500 mg/kg, of the extract intraperitoneally for 5 days and then received CCl4 (0.5 mL/kg, in olive oil). Liver injury was determined by serum biochemical markers such as alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), glutathione content of liver tissue and histopathological studies.ResultsThe phytochemical screening in Caspian saffron flowers indicated the presence of carotenoids, saponins, sterols, flavonoids, and tannins. The extract exhibited antioxidant activity in DPPH radical scavenging assay (IC50 171.5 µg/mL). Pre-treatment groups with the extract demonstrated significant decrease in serum levels of ALT, AST, and ALP (p<0.05) and concomitant increase in GSH content (p<0.05). Histopathological observation determined hepatocellular protective effects of the extract.ConclusionThe results represented the protective activity of the extract against oxidative hepatotoxicity.

HEPATOPROTECTIVE ROLE OF LOVASTATIN - PREGNANE X RECEPTOR AGONIST IN LITHOCHOLIC ACID INDUCED LIVER CHOLESTASIS DISEASED RATS

Objective: Pregnane X receptor (PXR), member of nuclear receptor family, an integral component of the body defence mechanism against chemical insult are expressed in the liver, gastrointestinal system &amp; lungs. Some studies have shown that the lovastatin is pregnane X receptor (PXR) activation effect.&#x0D; Methods: In the present study the hepatoprotective effect of lovastatin was investigated against lithocholic acid induced liver toxicity. Liver markers in serum and antioxidant enzymes in liver tissue were assessed by using standard procedures.&#x0D; Results: The level of liver marker (such as SGOT &amp; SGPT) and bilirubin were increased significantly (p&lt;0.05) and antioxidant enzyme (i.e. SOD, GSH and CAT) were significantly (p&lt;0.05) decrease in lithocholic acid treated groups as compared to control group. Lovastatin at doses of 0.1, 0.2, 0.3 mg/kg showed significantly (p&lt;0.05) decrease in the levels of liver marker (SGOT &amp; SGPT) and bilirubin as compared to the positive control group in both pre &amp; post treated models. The antioxidant enzymes such as Superoxide dismutase (SOD), Glutathione (GSH) and Catalase (CAT) content in liver tissue were significantly (p&lt;0.05) increase after administration of lovastatin at dose dependent manner in both pre &amp; post treated models.&#x0D; Conclusions: The results of the present study indicates that under the present experimental conditions, lovastatin showed hepatoprotective abilities against lithocholic acid induced hepatotoxicity in albino rat.&#x0D; Keywords: Hepatoprotection, lovastatin, Lithocholic Acid, Pregnane X Receptor

Chronic alcohol abuse may lead to high skin iron content, but not to hepatic siderosis

IntroductionIn forensic cases, ante mortem chronic alcohol abuse can be of central importance in clarifying circumstances of death. However, reliable markers of alcohol consumption, which are still available postmortem, are needed. In addition to medical history data which may not be always authentic, the determination of ethyl glucuronide (EtG) in hair as a promising parameter is of no value in cases of missing or cosmetically treated hair. On the other hand, there exist reports that iron ions accumulate in liver tissue (siderosis) during chronic, excessive alcohol consumption, which, therefore, may be useful to serve as alcohol abuse correlate. However, the influence of ethanol on iron stored in the liver has not been adequately investigated and the study situation appears to be inconsistent. AimsThe aim of the present study was to assess the suitability of assaying iron concentrations in liver and other tissues as postmortem alcoholism marker. MethodsThe iron concentration in tissue samples (liver, brain, skin, pancreas, spleen), vitreous fluid and blood taken during autopsy was analyzed by atomic absorption spectroscopy. The analytical method has been validated before. Cases were divided into two groups: chronic alcohol abusers and non-chronic alcohol consumers including total abstainers using ethyl glucuronide levels in hair as well as anamnestic data as criteria. ResultsNo elevated iron concentrations in the liver of chronic alcohol abusers were detected. Surprisingly, the iron concentration in skin tissue was found to be significantly higher in cases of chronic alcohol abuse, independent on whether fatty liver or liver cirrhosis was present (as diagnosedduring autopsy).In 18.5% of the cases, chronic alcohol abuse was not confirmed by the EtG concentration in hair. Thus, anamnestic data should not be overestimated. ConclusionThe general assumption that chronic alcohol abuse induces hepatic siderosis, i.e. high iron concentrations in liver tissue, has not been supported by results of the present study. However, there seems to exist a correlation between chronic alcohol abuse and high iron concentrations in the skin.

Medium-term dynamics of element concentrations in a sparid fish and its isopod parasite after the Prestige oil-spill: Shifting baselines?

Historically, the European Atlantic is probably the most important oil-spill hotspot worldwide. One of the most recent accidents occurred in 2002 when the oil-tanker Prestige sank over the Galician Bank causing two major oil-spills followed by several small leaks until March 2003. This resulted in contamination of virtually all types of marine habitat. Considering that parasites have proved to be good effect and accumulation bioindicators, the present study addresses the medium-term changes in trace element content after the Prestige oil-spill in a model host-parasite system, the bogue, Boops boops (Sparidae) and the isopod Ceratothoa oestroides. To our knowledge, this study is the first to address trace element concentrations in natural fish and parasite populations associated with the effects of an oil-spill. We observed that both test organisms examined, the host and the parasite, indicate a detectable change in the relative composition of trace element concentrations before and after the Prestige oil-spill. Multivariate analyses also indicated a differential response of the different tissues to the temporal sampling sequence. However, analyses of both host and parasite tissues supported the pattern of a gradual temporal transition to a state of relative trace element content distinctly departing from the pre-spill situation. Moreover, the parasite-host element accumulation ratios better depicted this temporal pattern. Additionally, changes in V concentrations in fish liver tissues and Ni concentrations in the parasite tissues suggest that this host-parasite system may be a useful tool to assess these two element contaminations linked to heavy fuel oil-spill.

The Impact of DJOS Surgery, a High Fat Diet and a Control Diet on the Enzymes of Glucose Metabolism in the Liver and Muscles of Sprague-Dawley Rats.

The prevalence of diabetes type 2 (T2DM) and obesity is growing exponentially and becoming a global public health problem. The enzymes of glucose metabolism play a role in the pathogenesis of insulin resistance and T2DM. A pathophysiological link between different dietary patterns, HFD, obesity, T2DM and the enzymes of glucose metabolism can be used as a potential target in therapeutic strategies for the treatment of obesity, and T2DM. The aim of this study was to measure the impact of DJOS bariatric surgery and different types of dietary patterns on glycogen synthase kinase 3 α (GSK-3α), glycogen phosphorylase (PYGM, PYGL), and phosphofructokinase (PFK-1) concentrations in liver and soleus muscle tissues of rats. After 8 weeks on a high-fat diet (HF) or control diet (CD), rats underwent duodenal-jejunal omega switch (DJOS) or SHAM (control) surgery. After surgery, for the next 8 weeks, half of DJOS/SHAM animals were kept on the same diet as before, and half had a changed diet. The concentrations of GSK-3α, PYGM, PYGL and PFK-1 were measured in the soleus muscles and livers of the Sprague-Dawley rats. The type of diet applied before/after surgery had stronger impact on levels of selected metabolic enzymes than DJOS or SHAM surgery. The impact of DJOS surgery was visible for GSK-3α and PYGL concentration in the liver but not in the soleus muscle tissue. The type of bariatric surgery had an impact on liver GSK-3α concentration in all studied groups except the CD/CD group, where the impact of diet was stronger. DJOS bariatric surgery influenced the level of PYGL in the livers of rats maintained on the CD/CD diet but not from other groups. The dietary patterns applied before and after bariatric surgery, had a stronger impact on enzymes’ concentrations than DJOS surgery, and the strong, deleterious effect of an HF was observed. A change of the diet per se showed a negative impact on the enzymes’ tissue concentration.

Ecotoxicoparasitology of mercury and trace elements in semi-aquatic mammals and their endoparasite communities

Many contaminants persist in the environment for decades or more, influencing ecosystem health. Environmental contamination with mercury (Hg) is a particular concern due to its ability to biomagnify in food webs and its lethal and sub-lethal effects in exposed organisms. Despite the known impacts of anthropogenic contamination, there remains a need for data on wildlife exposure to Hg and other contaminants, and the effects of exposure on wildlife health. The objectives of this study were to: 1) quantify differences in concentrations of mercury and other trace elements among three sympatric semiaquatic mammals of different assumed trophic position: North American river otter (Lontra canadensis), raccoon (Procyon lotor), and North American beaver (Castor canadensis), 2) compare trace element concentrations between animals captured on the Savannah River Site (SRS) in South Carolina, USA, where known inputs of Hg and other trace elements have occurred, and reference sites in South Carolina (SC) and Georgia (GA), USA, and 3) investigate the relationship between host trace element concentrations and endoparasite communities. River otters, beavers, and raccoons were sampled from the SRS, SC, and GA to quantify trace element concentrations in liver tissue and quantify endoparasite communities. Both species and sampling location were important factors determining hepatic trace element concentration, however, there was no consistent trend of elevated trace element concentrations among animals sampled on the SRS. Only Hg demonstrated biomagnification based on assumed trophic position, with river otters having the highest Hg concentrations among the sampled species. Additionally, the results suggest a possible relationship between host hepatic mercury concentration and endoparasite abundance, while hepatic selenium concentration may be related to endoparasite diversity. These findings further demonstrate how wildlife can accumulate anthropogenic contamination, although future research is needed to determine the mechanisms contributing to patterns observed between endoparasite communities and the contaminant concentrations of their mammalian hosts.

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

ABSTRACTHepatic 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.

LAYING PERFORMANCE, FAT DIGESTIBILITY AND LIVER CONDITION OF LAYING HENS SUPPLEMENTED WITH VITAMIN B12 OR BIOTIN AND/ OR BILE ACIDS IN DIET

For 8 weeks feeding trial, two hundred and seventy, 53 weeks old laying hens were used to investigate the effects of dietary supplementation of vitamin B12 or biotin and/or bile acids on performance, egg quality, fat digestibility and liver composition and histopathology. Birds were randomly divided into 6 groups (three replicates) and fed the experimental diets; group1 (G1) fed on the basal diet without additives as control while G2 and G3 supplemented with 0.02 ppm vitamin B12 and 0.3 mg biotin/kg diet respectively, groups 4-6 fed as the previous detailed design of G1- G3 with the addition of 400 g of dried bile acid (DBA) /tone feed. Biotin supplementation significantly (P˂0.05) increased body weight losses of laying hens, both vitamins significantly (P˂0.05) decreased daily feed intake (FI) and improved FCR. DBA addition alone or with biotin reduced these weight losses along with significant (P˂0.05) increase in daily FI. Vitamin B12 supplementation alone or with DBA increased average egg production (P˂0.05) while was reduced with biotin supplementation. Fat digestibility was non-significantly improved (P≥0.05) with both vitamins supplementation without or with DBA. Biotin significantly (P˂0.05) reduced the average yolk relative weight, which was increased when mixed with DBA, while significantly increased average egg albumin relative weight. Vitamin B12 or biotin addition without or with DBA non-significantly increased blood serum GOT and GPT activities, non-significantly reduced (P≥0.05) fat content of liver tissue (on dry matter or fresh basis) and serum lipid profile parameters except serum HDL concentration, was increased, with no histopathological changes in hepatic tissue. It could be summarized that vitamin B12 supplementation without or with DBA is recommended in layer diet as it gave the best performance, reduced serum lipid profile and improved fat digestibility and the hepatic health.Key words: layers; egg quality; liver; vitamin B12; biotin; bile acids

A multibiomarker approach for evaluating environmental contamination: Common carp (Cyprinus carpio) transplanted along a gradient of metal pollution

Environmental monitoring and risk assessment approaches which include a more holistic view on the effects of pollutants on biota are increasingly sought by regulators and policy makers. Therefore, caged carp juveniles (Cyprinus carpio) were transplanted for 7 weeks along a known Cd and Zn pollution gradient. Metal (Cu, Cd and Zn) accumulation in gill and liver tissue and effect biomarkers (growth, condition factor (CF), hepatosomatic index (HSI), oxygen consumption, swimming capacity, Na+/K+-ATPase activity (NKA) and metallothionein (MT) levels) were compared.Up to 10-fold higher cadmium concentrations were measured in the gills of the fish at the most polluted locations compared to the laboratory control fish. Similarly, cadmium concentrations in liver tissues of field-exposed fish were significantly higher than those measured in laboratory control fish. Cu and Zn concentrations in the gills were not significantly different between field-exposed and control organisms, whereas higher levels in liver tissues were measured in carps deployed in some locations. Effects on liver MT levels were up to 10 times greater for organisms exposed to the field, whereas no clear effect of the metal exposure on NKA in the gill tissue was observed. A decrease in muscle glycogen stores was observed for all organisms deployed in the field, while liver glycogen levels decreased only in fish exposed to two of the 5 sites compared to the laboratory control fish. Additionally, significant drops in liver protein- and lipid stores were observed. No effect on oxygen consumption rates and swimming capacity was observed. The CF and HSI of caged fish reflected the pollution gradient in the river and considerable loss of weight was observed for fish transplanted in the most polluted site. Overall, this active biomonitoring study successfully revealed differences in metal accumulation, physiological and organismal endpoints as a direct consequence of field exposure.

Protective effect of Schisandra chinensis total lignans on acute alcoholic-induced liver injury related to inhibiting CYP2E1 activation and activating the Nrf2/ARE signaling pathway

Schisandra chinensis (Turcz.) Baill., Schisandraceae, is a well-known traditional Chinese medicine used mainly as a recipe for hepatoprotection. Modern researches have revealed that the hepatoprotection is related to its lignans and crude polysaccharide. In this study, we examined the effect and mechanism of S. chinensis total lignans on the liver injury induced by alcoholic. S. chinensis total lignans were extracted with ethanol extraction. The liver index, alanine aminotransferase and aspartate aminotransferase levels in serum of the rat culture supernatant were examined. The malondialdehyde level and superoxide dismutase activities in serum and liver tissue, and triacylglyceride content in liver tissue were tested. Western blot was conducted to determine cytochrome P450 2E1 expression in liver tissue of rats. The results showed that S. chinensis total lignans administration significantly inhibited alcohol-induced liver injury. In exploring the underlying mechanisms of S. chinensis total lignans action, we found that it significantly decreased Alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (AKP), Glutamyl transpeptidase (γ-GT), reactive oxygen species (ROS) and malondialdehyde (MDA) level in livers/serum and inhibited the gene expression level of CYP2E1 in rat livers. The Nuclear factor erythroid-2 related factor 2 (Nrf2) gene expression and Nuclear factor erythroid-2 related factor 2 (Nrf2) protein nuclear transfer increased significantly, and significantly increased the expression of downstream target gene and protein heme oxygenase-1 (HO-1), Glutamate--cysteine ligase regulatory subunit (GCLM), NAD(P)H:quinine oxidoreductase 1 (NQO1). Moreover, S. chinensis total lignans decreased production of pro-inflammatory markers including Tumor Necrosis Factor-α (TNF-α), Interleukin-1beta (IL-1β) and Interleukin-6 (IL-6). In conclusion, these results suggested that the inhibition of alcohol-induced liver injury by S. chinensis total lignans is associated with its ability to inhibiting CYP2E1 activation and activating the Nrf2/Antioxidant response element(ARE) signaling pathway.

Quantification and genetic diversity of Hepatitis E virus in wild boar (Sus scrofa) hunted for domestic consumption in Central Italy

Hepatitis E is an emerging disease in industrialized countries. The food-borne transmission of hepatitis E virus (HEV) is associated principally with products derived from the domestic pig, the wild boar, and deer; however, few quantitative data are available on HEV loads in animals used in food production. This study assessed HEV occurrence, viral load and genetic variability in wild boar hunted for domestic consumption in the district of Viterbo (Central Italy) where high anti-HEV IgG seroprevalence values are reported in humans. A total of 332 liver and 69 intestine samples were obtained from wild boar hunted between 2011 and 2014. The liver tissue in 54 of the animals (16.3%) was HEV-positive. Viral loads in quantifiable liver samples (n = 29) ranged between 3.2 × 102 and 3.8 × 105 genome copies (g.c.)/g with a mean value of 1.85 × 104 g.c./g. A statistically significant positive correlation was found between viral concentration in liver and intestinal tissue, though mean viral load in the intestines was lower (3.13 × 103 g.c./g). Twenty-six samples were characterized molecularly as genotype 3 (G3) and four subtypes (a, c, f and l) were detected. Finally, twelve samples with near identical sequences were identified as G3 but could not be assigned to any of the known subtypes, and could therefore represent a potentially new subtype.

Selenium deficiency induced an inflammatory response by the HSP60 - TLR2-MAPKs signalling pathway in the liver of carp

Selenium (Se) is one of the essential trace elements for immune regulation and antioxidant systems in fish growth. The dietary Se plays an important role in immune regulation and inflammation by regulating HSPs and TLRs in liver of many animals. The liver is an important digestive organ in carp. Liver damage can seriously affect the growth and survival of carp. This study was conducted to determine whether Se regulated liver inflammation by affecting HSPs-TLR2 signalling and the potential mechanisms of action in common carp. The gene was analysed by qPCR. The proteins of inflammatory factors were detected by ELISA. The others proteins were analysed by Western blot. The results indicated the Se concentrations in blood and liver tissues were significantly influenced by dietary Se. The Se deficiency increased the expression of HSP60 and TLR2 and the secretion of the proinflammatory factor TNF-α, IL-1β and IL-6, induced a low secretion of the anti-inflammatory TGF-β, but the Se supplements could transform these events. Further research showed that with the dose-dependently decrease of Se, the HSP60 expressions were increased, and the MAPKs pathway were significantly activated by the phosphorylation of p38, JNK and ERK in liver tissue and cell. The results provide evidence that Se deficiency induced and exacerbated inflammatory injury to the liver through the HSP60 and TLR2-MAPKs signalling pathways in carp.