Influence of Dietary Metformin on the Growth Performance and Plasma Concentrations of Amino Acids and Advanced Glycation End Products in Two Types of Chickens

Glycation is a non-enzymatic reaction inducing the bonding of glucose to amino acids and proteins. Glycated amino acids are not useful for protein synthesis, suggesting that glycation reduces the utilization of amino acids. Metformin (MF) is well known as a therapeutic drug for type II diabetes that inhibits glycation. It is possible that treatment with MF raises the utilization of amino acids by the inhibition of glycation, thereby improving the growth performance of chickens. In the present study, therefore, we investigated the influence of dietary MF on the growth performance, and plasma concentrations of free amino acids and N^ε-(Carboxymethyl)lysine (CML), which is an advanced glycation end product, in layer (Experiment 1) and broiler (Experiment 2) chickens. From 7 d of age, chicks were allowed free access to one of the experimental diets containing MF at 3 supplementation levels (0, 150, and 300 mg/kg diet) for 14 days. Body weight and feed intake were measured every week. At the end of the experiments, blood and breast muscle (M. pectoralis major) were collected for further analysis. Dietary MF did not affect weight gain, feed intake, or feed efficiency in both layer and broiler chickens. Dietary MF at the level of 150 mg/kg diet increased breast muscle weight in both layer and broiler chickens. Dietary MF increased plasma concentrations of branched chain amino acids and decreased concentrations of CML in layer chickens, although it did not affect plasma concentrations of glucose. The present study suggested that dietary MF might have the potency to increase breast muscle weight of layer chickens with an increment in plasma concentrations of branched-chain amino acids.     

Advanced glycation end products and sorbitol in blood from differently compensated diabetic dogs

Canine diabetes mellitus (DM) is a common metabolic disorder with long term complications, most of which are caused by glycosylation of structural proteins, decreases in antioxidant concentrations, altered osmotic balance and hypoxia due to impaired oxygen transport. Previous studies have demonstrated that under hyperglycemic conditions canine erythrocytes undergo swelling, probably due to activation of the polyol pathway. The present work aimed to assess the plasma concentration of advanced glycation end (AGE) products, stable Amadori-products generated by non-enzymatic glycosylation of proteins and the intracellular concentration of sorbitol, produced by the activation of polyol pathway in 34 blood samples from diabetic dogs and in 14 controls. AGE products were significantly higher (p<0.01) in plasma from diabetic dogs compared with control animals. The sorbitol concentration in erythrocytes was also significantly higher in diabetic dogs and, in particular, in poorly compensated animals and in dogs with ketonuria. In five cases that were analysed before and after clinical improvement, sorbitol concentration was found to correlate with improvement. These results suggest that non-specific glycosylation is increased and that the polyol pathway is activated in diabetic dogs in a manner that is proportionate to the severity of disease. Moreover, the concentration of AGE products and sorbitol may be useful for monitoring the onset of diabetic complications and assessing the most appropriate therapeutic approaches for management of canine DM.     

Estimation of concentration of radionuclides in skeletal muscle from blood,based on the data from abandoned animals in Fukushima

The damage caused by the earthquake on 11 March, 2011 resulted in a serious nuclear accident in Japan. Due to the damage to the Fukushima Daiichi Nuclear Power Plant (FNPP), large amounts of radioactive substances were released into the environment. In particular, one of the largest safety concerns is radioactive cesium (¹³⁴Cs and ¹³⁷Cs). Due to the FNPP nuclear accident, a 20 km area was restricted from human activity, and various types of domestic animals were left in the zone. We collected the organs and tissues from sacrificed animals to obtain scientific data to evaluate the internal deposition of radioactive compounds. At first, we found there is a strong correlation between blood ¹³⁷Cs and organ ¹³⁷Cs with data from 44 cattle, indicating that skeletal muscle is the target organ of deposition of radioactive cesium. Second, we analyzed the relationship between blood ¹³⁷Cs and muscle ¹³⁷Cs within relatively lower radioactive concentration, suggesting that estimation of concentration of ¹³⁷Cs is possible from blood concentration of ¹³⁷Cs. Finally, we developed computer software to estimate the muscle ¹³⁷Cs concentration from blood samples. Our study contributes to the food safety of livestock products.     

Antioxidation status and histidine dipeptides content in broiler blood and muscles depending on protein sources in feed

One‐day‐old chickens were fed mixtures containing different raw materials (fish by‐products meal, porcine blood cells meal, blood meal, wheat gluten, fodder yeast), as a source of histidine and β‐alanine – components of carnosine. Control birds were administered a feed mixture, in which soy bean meal was the main protein source. The bodyweight, feed consumption and conversion, antioxidant characteristics and histidine dipeptides content in blood and muscles, and also amino acid composition of chicken meat on day 34 post‐hatch were recorded. The best (p < 0.05) performance and feed conversion were observed in chickens fed mixture containing porcine blood cells meal. In blood plasma of control chickens, a significantly (p < 0.01) higher ability to scavenge DPPH radicals was found. However, the highest catalase activity in erythrocytes was determined in chickens fed mixtures with blood by‐products. Insignificant differences in both carnosine and anserine levels in plasma between treatments were noted. Breast muscles from control birds were characterized by lower activity of glutathione peroxidase (GPx), superoxide dismutase (SOD) and catalase (CAT) (p < 0.05; p < 0.01), than those from chickens fed blood by‐products. Improved ability to reduce ferric ions (FRAP) (p < 0.01) and carnosine content in meat from chickens fed blood cell meal were recorded. No direct relations between amino acids content in feed mixtures and in meat were observed.     

Assessing serum metabolite profiles as predictors for feed efficiency in broiler chickens reared at geographically distant locations

1. The objective of this study was to investigate differences in growth performance, serum intermediary metabolites, acute-phase proteins and white blood cells in low, medium and high–residual feed intake (RFI) chickens. It was also assessed if the environment affects the feed efficiency (FE) and FE-related performance and serum profiles of chickens.

2. Individual body weight (BW) and feed intake (FI) were recorded from d 7 of life. At 5 weeks of age, female and male broiler chickens (Cobb 500) were selected according to their RFI (L1: Austria; L2: UK; n = 9/RFI group, sex and locatity -45on) and blood samples were collected.

3. Chickens at L1 had similar FI but a 15% higher BW gain compared to chickens at L2. The RFI values of female chickens were ?231, 8 and 215 g and those of male chickens ?197, 0 and 267 g for low, medium and high RFI, respectively.

4. Location affected serum glucose, urea, cholesterol, non-esterified fatty acids (NEFA) and ovotransferrin in females, and serum glucose and triglycerides in male chickens. Serum uric acid and NEFA linearly increased from low to high RFI in females, whereas in males, cholesterol showed the same linear response from low to high RFI. Serum alpha-1-acid glycoprotein and blood heterophil-to-lymphocyte ratio linearly increased by 35% and 68%, respectively, from low to high RFI but only in male chickens at L1.

5. Regression analysis showed significant positive relationships between RFI and serum uric acid (R² = 0.49) and cholesterol (R² = 0.13).

6. It was concluded that RFI-related variation in serum metabolites of chickens was largely similar for the two environments and that serum metabolite patterns could be used to predict RFI in chickens.     

Glycated Tryptophan PHP-THβC Incorporated into Various Chicken Embryonic Cells Constitutes Cellular Proteins

Glycation is a non-enzymatic reaction, and amino acids are glycated by glucose in vivo. Tryptophan is glycated with glucose to form two types of glycated compounds, tryptophan-Amadori product and (1R, 3S)-1-(D-gluco-1, 2, 3, 4, 5-pentahydroxypentyl)-1, 2, 3, 4-tetrahydro-β-carboline-3-carboxylic acid (PHP-THβC). Although PHP-THβC can be incorporated into various chicken embryonic cells, the mechanism of its incorporation into intracellular fluids has not been clarified. In this study, we examined whether PHP-THβC once incorporated into various chicken embryonic cells can combine with proteins. Embryonic cells from the breast muscle, liver, spleen, kidney, proventriculus, gizzard, and skin were prepared and ³H-PHP-THβC was added to the culture medium at final concentrations of 0, 200, 400, 600, and 800 µM to examine the incorporation of PHP-THβC. After 18 h of incubation, radioactivity was measured in the whole-cell and protein fractions of the chicken embryonic cells. As PHP-THβC concentration increased from 0 to 600 µM, its accumulation in the whole-cell fractions of all types of chicken embryonic cells linearly increased and reached the maximum level. The saturated PHP-THβC accumulation in the whole-cell fractions suggests that PHP-THβC could be incorporated into intracellular fluids across cellular membranes by some transporter proteins. As PHP-THβC concentration increased from 0 to 800 µM, its accumulation in the protein fractions of all types of chicken embryonic cells increased in a linear manner and reached a maximum level in the 800 µM PHPTHβC treatment group. This is the first study to indicate that a part of PHP-THβC incorporated into the whole-cell fraction was detected in the protein fraction of various chicken embryonic cells.     

STATIC AND DYNAMIC 18FDG‐PET IN NORMAL HISPANIOLAN AMAZON PARROTS (AMAZONA VENTRALIS)

Positron emission tomography (PET) is often used to stage and monitor human cancer and has recently been used in a similar fashion in veterinary medicine. The most commonly used radiopharmaceutical is 2‐Deoxy‐2‐[¹⁸F]‐Fluoro‐d ‐glucose (¹⁸F‐FDG), which is concentrated and trapped within cells that use glucose as their energy substrate. We characterized the normal distribution of ¹⁸F‐FDG in 10 healthy Hispaniolan Amazon parrots (Amazona ventralis) by performing whole body PET scans at steady state, 60 min after injection. Significant variability was found in the intestinal activity. Avian species are known to reflux fluid and electrolytes from their cloaca into their colon. To evaluate reflux as the cause of variability in intestinal distribution of ¹⁸F‐FDG, dynamic PET scans were performed on the coelomic cavity of six Hispaniolan Amazon parrots from time 0 to 60 min postinjection of radiotracer. Reflux of radioactive material from the cloaca into the colon occurred in all birds to varying degrees and occurred before 60 min. To evaluate the intestinal tract of clinical avian patients, dynamic scans must be performed starting immediately after injection so that increased radioactivity due to metabolism or hypermetabolic lesions such as cancer can be differentiated from increased radioactivity due to reflux of fluid from the cloaca.     

Turnover of Se in adequately fed chickens using Se‐75 as a tracer

Inorganic selenium (Se) in the form of selenite is applied to livestock to avoid Se deficiency. Selenite is, however, an artificial Se source in diets of unsupplemented chickens. It is therefore hypothesized that organic Se sources, such as Se‐enriched yeast and wheat, could be a more suitable Se supply in animal nutrition, although information on the transition of Se from organic Se sources in fast‐growing chickens is scarce. In this work, chickens were fed a low Se diet (0.27 ± 0.01 mg Se/kg, Se‐enriched yeast) until 20 days of age, after which the Se concentration was increased to maximum concentration allowed by the poultry industry in Europe (0.5 p.p.m. Se). At the same time, a daily contribution of carrier‐free ⁷⁵Se tracer from labelled wheat was administered from day 20 to 27. The chickens showed S and Se homeostasis, as the concentration of S and Se in liver, blood or kidney remained about constant, and steady state of S and Se in the other organs was reached 1 day after the diet shift. The uptake of ⁷⁵Se was readily seen in all organs. After 1 week, the depuration of the ⁷⁵Se tracer was followed, and biological half‐lives and retention in individual organs were determined. The shortest biological half‐lives were observed in major metabolic organs, the liver, kidney and pancreas with half‐lives close to 4 days. There was a significant (p < 0.05) uptake in lung, brain and muscle that reached steady state when the administration of ⁷⁵Se was terminated. The half‐life of ⁷⁵Se in heart was 9 days and 7 days in blood. The longest half‐lives were observed in muscle (12 days), brain and lungs (13 days). All half‐lives were shorter than in Se deplete animals.     

The effects of the supplementation of multi‐strain probiotics on intestinal microbiota,metabolites and inflammation of young SPF chickens challenged with Salmonella enterica subsp. enterica

This study assessed the effect of probiotics on cecal microbiota, cecal short‐chain fatty acids (SCFAs), and the gene expression of cytokines in young specific‐pathogen‐free (SPF) chickens infected with S. enterica subsp. enterica. One‐day‐old SPF chickens (n = 105) were randomly assigned to one of the three treatment groups: control (Cont) group, Salmonella‐infected (Sal) group, and a Salmonella‐infected group treated with multi‐strain probiotics (ProSal group). All chickens except those in the Cont group were challenged orally with 1 × 10⁸ cfu/ml of Salmonella 4 days after hatching. Chickens in the Sal group exhibited more abundance of Proteobacteria than those in the Cont and ProSal groups. At the genus level, chickens in ProSal group exhibited increased numbers of Lactobacillus and Oscillospira compared with those in the other groups. Chickens in the ProSal group exhibited a significant increase of cecal SCFAs compared with chickens in the Sal group. Chickens in the ProSal group exhibited increased gene expression of anti‐inflammatory cytokines, IL‐10 and TGF‐β4, and decreased expression of the proinflammatory cytokine, IFN‐γ, in the cecal tonsil compared with those in the Sal group. The results of this study indicated that the administration of probiotics can modulate microbiota, SCFAs, and immunomodulatory activity in SPF chickens.     

Urinary excretion of advanced glycation end products in dogs and cats

The present study was conducted with privately owned dogs and cats to investigate whether a relationship exists between the dietary AGEs and the urinary excretion of AGEs, as indication of possible effective absorption of those compounds in the intestinal tract of pet carnivores. For this purpose, data were collected from both raw fed and dry processed food (DPF) fed to dogs and cats, through spot urine sampling and questionnaires. Raw pet food (RF, low in AGE diets) was fed as a primary food source to 29 dogs and DPF to 28 dogs. Cats were categorized into 3 groups, which were RF (n = 15), DPF (n = 14) and dry and wet processed pet food (DWF, n = 25). Urinary-free carboxymethyllysine (CML), carboxyethyllysine (CEL) and lysinoalanine (LAL) were analysed using ultrahigh-performance liquid chromatography (UHPLC)—mass spectrometry, and were standardized for variable urine concentration by expressing the AGE concentrations as a ratio to urine creatinine (Ucr) concentration (µg/µmol Ucr). Urinary excretion of CML, CEL and LAL in dogs fed with DPF was 2.03, 2.14 and 3 times higher compared to dogs fed with RF (p < .005). Similar to the dogs, a significant difference in CML:Ucr, CEL:Ucr and LAL:Ucr between the three diet groups was observed in cats (p-overall < 0.005, ANOVA), in which the RF fed group excreted less AGEs than the other groups. Linear regression coefficients and SE of CML:Ucr, CEL:Ucr and LAL:Ucr showed that body weight and neuter status were significantly correlated with CML and CEL excretion, but not to LAL excretion. Our results revealed a significant correlation between dietary AGEs and urinary excretion of free CML, CEL and LAL, and also showed that endogenous formation of these AGEs occurs in both dogs and cats under physiological conditions.     

Variability in nutrient composition and in vitro crude protein digestibility of 16 microalgae products

The chemical composition of 16 microalgae products of four genera, Arthrospira (n = 2), Chlorella (n = 8), Nannochloropsis (n = 4) and Phaeodactylum (n = 2), was assayed to evaluate the intra‐ and inter‐genera variation of nutrient profiles of commercial microalgae products. Crude protein was the main component in all genera, followed by ether extract and crude ash. Mean crude protein concentrations were 690, 502, 431 and 446 g/kg dry matter, and mean ether extract concentrations were 63, 157, 188 and 113 g/kg dry matter for Arthrospira, Chlorella, Nannochloropsis and Phaeodactylum respectively. However, there was considerable inter‐ and intra‐genera variation. The concentration of α‐linked glucose was low (0–143 g/kg dry matter). There was high variation between and within genera in the crude ash concentration (22–237 g/kg dry matter), which was also observed for the mineral composition. In contrast to the crude protein concentration, the amino acid composition of the protein (g amino acid/16 g N) was less variable. The investigated samples possessed high concentrations of Glx, Asx and Leu, and low concentrations of Cys and Met. The mean concentration of non‐protein nitrogen compounds was highest in Phaeodactylum (110 g/kg dry matter) and lowest in Nannochloropsis (47 g/kg dry matter) products, and as with proximate nutrients, high variability between and within genera was observed. In vitro crude protein digestibility varied between 54% (non‐cell‐disrupted Nannochloropsis) and 84% (cell‐disrupted Chlorella). Inositol phosphate isomers were not detectable in any sample (concentration <1 μmol/g dry matter). The predominant fatty acids were C16:0 in Arthrospira products, C18:2 n‐6+ C19:1 t7 and C18:3 n‐3 in Chlorella products, and C20:5 n‐3 in Nannochloropsis and Phaeodactylum products; however, the relative proportions of fatty acids varied within genera. Commercially available microalgae products appear to be valuable alternative food and feed products. However, because of the high variability in nutrient profiles, attention should be given to the analytical characterization of the products.     

Binding of glycated ovocystatin to rat renal brush border membranes

Glycated proteins are considered as one of the factors involved in the pathogenesis of diabetic complications, including nephropathy. These proteins are formed endogenously under conditions of hyperglycemia, as well as being provided with food containing sugars, which was subjected to high temperature. Examples are egg products. One of the proteins found in eggs in a relatively high concentration is chicken cystatin (ovocystatin). It is now believed that some proteins can passage the intestinal epithelium by transcytosis directly into the bloodstream. Thus, glycated protein present in food can be an additional source of glycotoxins. The aim of this study was to compare the affinity of native and glycated cystatin to the brush border membranes of rat kidney. Kinetic analysis was performed with surface plasmon resonance technique using sensor chip L1. Dissociation constants for native and glycated cystatin (K_d) were 2.76 μmol/L and 3.82 μmol/L, respectively. The results of our study indicate that glycation only slightly affects binding of cystatin to brush border membranes. This suggests that glycated cystatin and other glycated proteins may also be efficiently taken up in the kidney proximal tubule. The observation may be important for understanding the mechanisms involved in the development of diabetic nephropathy.     

Cesium radioactivity in peripheral blood is linearly correlated to that in skeletal muscle: Analyses of cattle within the evacuation zone of the Fukushima Daiichi Nuclear Power Plant

The accident at the Fukushima Daiichi Nuclear Power Plant (FNPP) released a large amount of radioactive substances into the environment. Furthermore, beef contaminated with radioactive cesium above the 500 Bq/kg safety standard was circulated in the food chain in 2011. Japanese consumers remain concerned about the safety of radioactively contaminated food. In our previous study, we detected a linear correlation between radioactive cesium (¹³⁷Cs) activity in blood and muscle around 500 to 2500 Bq/kg in cattle. However, it was unclear whether the correlation was maintained at a lower radioactivity close to the current safety standard of 100 Bq/kg. In this study, we evaluated 17 cattle in the FNPP evacuation zone that had a ¹³⁷Cs blood level less than 10 Bq/kg. The results showed a linear correlation between blood ¹³⁷Cs and muscle ¹³⁷Cs (Y = 28.0X, R² = 0.590) at low radioactivity concentration, indicating that cesium radioactivity in the muscle can be estimated from blood radioactivity. This technique would be useful in detecting high‐risk cattle before they enter the market, and will contribute to food safety.     

Ileal amino-acid digestibility of wheat, autoclaved wheat and spaghetti by-products for broiler chicks

An experiment was conducted to determine true and apparent ileal amino‐acid digestibility of a native cultivar of wheat (Mahdavi), autoclaved wheat (120°C for 30 min) and spaghetti by‐products available in Iran. One hundred 21‐day‐old broiler chickens were fed a standard corn–soybean meal starter diet from day 0 to 28 post hatch. At 28 days 80 chicks were distributed according to possessing very nearly the same average bodyweight to 20 wire cages. The experimental units were allocated at random to five dietary treatments with four replicates per treatment. The basal diet contained corn and soybean meal as the major ingredients. Three test diets were formulated containing wheat, autoclaved wheat and spaghetti by‐products as the sole source of dietary protein and each test diet was combined with a basal diet 50:50 on a weight basis to form three assay diets. The apparent and true digestibility of amino acid in the test ingredients were estimated from those in the assay diets basal/test diet mixtures, using the difference method. The apparent and true amino‐acid digestibility of the test ingredients were significantly different (P < 0.01). Autoclaving of wheat increase its amino‐acid digestibility (P < 0.01). Among the test ingredients, the average ileal amino‐acid digestibility of spaghetti by‐products was higher than wheat and autoclaved wheat but it was lower than values for the basal diet (P < 0.01).     

Technical Note: In vivo estimation of lipogenesis using a bolus injection of [U-13C]glucose in pigs

The use of radioactive isotopes to measure de novo lipogenesis in pigs has been well established. Different from radioactive isotopes, stable isotopes present little or no risk to human and animal subjects. Therefore, the objective of this study was to adapt the method of bolus injection of radioactive glucose (¹⁴C) to use ¹³C-labeled glucose to estimate de novo lipogenesis in finishing pigs. Five vein-catheterized gilts received 3.0 kg/d of a commercial diet for 2 wk. On the last day, the pigs received a bolus injection of [U-¹³C]glucose (12 mg/kg body weight). A serial of blood samples was taken for 4 h to determine the glucose rate of disappearance (R_d) from plasma glucose isotopic enrichment (IE). The ¹³C IE of lipids was determined from adipose tissue biopsies collected at 1, 2, and 3 h after the bolus injection and from adipose tissue collected after pig euthanasia 4 h after the bolus. Lipogenesis was estimated from the incorporation of ¹³C from glucose into adipose tissue lipids. Glucose R_d, estimated using a double-exponential function, averaged 5.4 ± 1.4 mmol/min. The IE of lipids increased linearly during the 4 h following the bolus injection (P < 0.05). The rate of incorporation of glucose into lipids, estimating lipogenesis, averaged 9.0 µg glucose/(min × g of lipids) 4 h after the bolus injection. In conclusion, the in vivo method using a bolus injection of [U-¹³C]glucose allows a successful estimation of de novo lipogenesis in finishing pigs.     

Effect of nutrient intake on intramuscular glucose metabolism during the early growth stage in cross‐bred steers (Japanese Black male × Holstein female)

The objective was to investigate the impact of nutrient intake during the early growth period on the expression of glucose metabolism‐related genes in skeletal muscle of cross‐bred cattle. From 1.5 to 5 months of age, group H (n = 7) animals were intensively fed a high‐protein and low‐fat milk replacer [crude protein (CP) 28%; ether extracts (EE) 18%; max: 2.0 kg, 12 l/day], and group R (n = 7) animals were fed a restricted amount of normal milk replacer (CP 25%; EE 23%; max 0.5 kg, 4 l/day). From 6 to 10 months of age, group H cattle were fed a high‐nutrition total mixed ration mainly prepared from grain feed, and group R cattle were fed only roughage. Blood samples were taken from each animal at three biopsy times (1.5, 5 and 10 months of age), and the blood plasma concentration of glucose and insulin was analysed. In glucose concentration, there were no significant differences; however, the concentrations of insulin were higher in group H than in group R at 5 and 10 months of age. Muscle samples were taken by biopsy from longissimus thoracis muscle (LT) at 1.5, 5 and 10 months of age. We analysed mRNA expression levels using the quantitative real‐time polymerase chain reaction (PCR) assay for glucose transporters (GLUT1 and GLUT4), insulin receptor, phosphatidylinositol 3‐kinase (PI‐3K), protein kinase B (PKB, also known as Akt), hexokinase 1 (HK1) and tumour necrosis factor alpha (TNFα). Although no differences were detected at 1.5 and 5 months of age, at 10 months of age, GLUT1, HK1 and TNFα mRNA expression levels were significantly higher in group H than in group R. These results suggested Glut1 that affects insulin‐independently mediated glucose uptake was more responsive to improved nutrition during early growth stage than GLUT4 that insulin‐dependently mediated glucose uptake in LT of cattle.     

Advanced glycation endproducts in horses with insulin-induced laminitis

Advanced glycation endproducts (AGEs) have been implicated in the pathogenesis of cancer, inflammatory conditions and diabetic complications. An interaction of AGEs with their receptor (RAGE) results in increased release of pro-inflammatory cytokines and reactive oxygen species (ROS), causing damage to susceptible tissues. Laminitis, a debilitating foot condition of horses, occurs in association with endocrine dysfunction and the potential involvement of AGE and RAGE in the pathogenesis of the disease has not been previously investigated. Glucose transport in lamellar tissue is thought to be largely insulin-independent (GLUT-1), which may make the lamellae susceptible to protein glycosylation and oxidative stress during periods of increased glucose metabolism. Archived lamellar tissue from horses with insulin-induced laminitis (n=4), normal control horses (n=4) and horses in the developmental stages (6h, 12h and 24h) of the disease (n=12) was assessed for AGE accumulation and the presence of oxidative protein damage and cellular lipid peroxidation. The equine-specific RAGE gene was identified in lamellar tissue, sequenced and is now available on GenBank. Lamellar glucose transporter (GLUT-1 and GLUT-4) gene expression was assessed quantitatively with qRT-PCR in laminitic and control horses and horses in the mid-developmental time-point (24 h) of the disease. Significant AGE accumulation had occurred by the onset of insulin-induced laminitis (48 h) but not at earlier time-points, or in control horses. Evidence of oxidative stress was not found in any group. The equine-specific RAGE gene was not expressed differently in treated and control animals, nor was the insulin-dependent glucose transporter GLUT-4. However, the glucose transporter GLUT-1 was increased in lamellar tissue in the developmental stages of insulin-induced laminitis compared to control horses and the insulin-independent nature of the lamellae may facilitate AGE formation. However, due to the lack of AGE accumulation during disease development and a failure to detect an increase in ROS or upregulation of RAGE, it appears unlikely that oxidative stress and protein glycosylation play a central role in the pathogenesis of acute, insulin-induced laminitis.     

Comparative development of digestive organs,intestinal disaccharidases and some blood metabolites in broiler and layer‐type chicks after hatching

1. Body weight, digestive organ weights, and activities of disaccharidases (maltase and saccharase) activities were determined from day of hatch to 21 d of age in meat‐ and egg‐type chickens. Blood plasma was analysed for enzyme activities and metabolite concentration.

2. In meat‐type chickens food intake and growth rate were about 3‐fold those in egg‐type chickens. Food efficiency was superior in meat‐type chickens throughout the experimental period.

3. Meat‐type chickens hatched with disaccharidase activities exceeding those found in their egg‐type counterparts 2‐ to 5‐fold. From 7 d of age on, this trend reversed, i.e. activity was much higher in egg‐type than in meat‐type chickens.

4. Blood plasma amylase activity increased gradually in meat‐type chickens and was higher than in egg‐type chickens to 14 d of age. No breed differences were observed for alkaline phosphatase or lactate dehydrogenase activities during the experimental period.

5. Blood plasma concentrations of total protein, albumin, glucose, and calcium, were lower in meat than in egg‐type chickens.     

H5N1亚型禽流感病毒感染海兰白鸡模型的建立

为建立H5N1亚型禽流感病毒感染海兰白鸡模型,本研究选取1株鹅源H5N1高致病性禽流感病毒A/goose/guangdong/1/96(H5N1)(简称GD1/96),测定其对4周龄海兰白鸡的半数致死量.感染模型试验中,将30只4周龄海兰白鸡随机分成3组,每组10只,5只直接感染,5只同居,试验组设置一个重复,将病毒液稀释至10^4.5EID₅₀,滴鼻、点眼各0.1 mL,对照组接种PBS,感染后24 h放入同居鸡;感染后连续观察14 d,记录死亡时间,每天采集咽喉拭子和泄殖腔拭子;感染组和同居组第3、5 天各剖解3只鸡,采集气管、肺脏、脑、脾脏、肾脏和十二指肠,进行病毒分离;qRT-PCR法分析感染组和同居组第3、5 天鸡肺组织中IFN-α和TNF-α的相对表达量.结果显示,GD1/96株的鸡胚半数感染量(EID₅₀)为10^-8.167/0.1 mL,对4周龄海兰白鸡的半数致死量为10^4.5 EID₅₀.感染模型试验结果显示,以10^4.5EID₅₀的攻毒剂量感染海兰白鸡,感染组鸡在感染后8 d全部死亡;在感染和同居3 d后,各组鸡的咽喉拭子和泄殖腔拭子均可检测到病毒;感染和同居后第3、5 天,各组鸡的6种组织中均可分离到高滴度的病毒;IFN-α和TNF-α在感染组和同居组的鸡肺脏组织中的表达量均显著增加(P <0.05).本试验建立了海兰白鸡的H5N1亚型禽流感病毒感染模型,为H5N1亚型禽流感病毒的致病机理及表达抗流感基因转基因鸡的研究奠定了基础.