Persistent hypoglycemia is induced by tolbutamide administration in broiler chickens fed a low-carbohydrate diet

With a view to gaining an insight into the regulatory mechanism of blood glucose concentrations specific to the chicken, an experimental induction of hypoglycemia was conducted by single or sequential administration of tolbutamide in broiler chickens fed a standard or low-carbohydrate diet. A single dosing of tolbutamide at levels of 25-200 mg/kg body weight decreased plasma glucose concentrations for 2 to 8 h after the dosing in chickens fed either diet. No significant rise in plasma insulin concentration was observed for 2 to 24 h after the single dosing of tolbutamide in chickens on either diet, with the exception of a significant rise when chickens on the standard diet received 100 mg tolbutamide. However, a transient increase of plasma insulin concentration was observed only in the 20 min immediately after the single dosing. Persistent hypoglycemia that was sustained for 5 days, with no significant changes in plasma insulin concentration, was induced by sequential dosing (3 times per day for 5 days, every 8 h) of tolbutamide (100 and 200 mg/kg body weight) in chickens fed the low-carbohydrate diet. In these chickens, the consistently low concentration of plasma glucose, with small diurnal variations, was evidenced by the determination of plasma glucose every 3 h in day 4/5 of the tolbutamide dosing. In chickens fed the standard diet, on the other hand, the low plasma glucose concentrations for 5 days were accompanied by significant diurnal fluctuations. Chickens with persistent hypoglycemia showed slight decreases in plasma non-esterified fatty acids (NEFA) concentration and only slight changes in blood D-3-hydroxybutyrate (3HB) concentration. The present study shows that the persistent hypoglycemia with normoinsulinemia, in the main, is induced by tolbutamide dosing in chickens fed a low-carbohydrate diet, and that the blood concentrations of NEFA and 3HB, alternatives of energy source in animals, are only slightly changed or not at all in hypoglycemic chickens.     

Kisspeptin,c‐Fos and CRFR Type 2 Co‐expression in the Hypothalamus after Insulin‐Induced Hypoglycaemia

Normal reproductive function is dependent upon availability of glucose and insulin‐induced hypoglycaemia is a metabolic stressor known to disrupt the ovine oestrous cycle. We have recently shown that IIH has the ability to delay the LH surge of intact ewes. In the present study, we examined brain tissue to determine: (i) which hypothalamic regions are activated with respect to IIH and (ii) the effect of IIH on kisspeptin cell activation and CRFR type 2 immunoreactivity, all of which may be involved in disruptive mechanisms. Follicular phases were synchronized with progesterone vaginal pessaries and at 28 h after progesterone withdrawal (PW), animals received saline (n = 6) or insulin (4 IU/kg; n = 5) and were subsequently killed at 31 h after PW (i.e., 3 h after insulin administration). Peripheral hormone concentrations were evaluated, and hypothalamic sections were immunostained for either kisspeptin and c‐Fos (a marker of neuronal activation) or CRFR type 2. Within 3 h of treatment, cortisol concentrations had increased whereas plasma oestradiol concentrations decreased in peripheral plasma (p < 0.05 for both). In the arcuate nucleus (ARC), insulin‐treated ewes had an increased expression of c‐Fos. Furthermore, the percentage of kisspeptin cells co‐expressing c‐Fos increased in the ARC (from 11 to 51%; p < 0.05), but there was no change in the medial pre‐optic area (mPOA; 14 vs 19%). CRFR type 2 expression in the lower part of the ARC and the median eminence was not altered by insulin treatment. Thus, disruption of the LH surge after IIH in the follicular phase is not associated with decreased kisspeptin cell activation or an increase in CRFR type 2 in the ARC but may involve other cell types located in the ARC nucleus which are activated in response to IIH.     

Time-action profiles of insulin detemir in normal and diabetic dogs

Insulin detemir is the first member of a new class of long-acting soluble insulin analogues capable of maintaining the basal level of insulin in humans. In this preliminary study, we investigated the time-action profiles of insulin detemir in normal and diabetic dogs since the use of insulin detemir in canines has yet to be determined. Eight animals were used in our study (three normal and five insulin dependent diabetic dogs). Time-action profiles of insulin detemir were monitored in normal dogs using an artificial pancreas apparatus under euglycemic condition. Blood sampling was performed at 2 h intervals post feeding, with insulin administration, in insulin dependent diabetic dogs. Time-action profiles of insulin detemir, in normal dogs, demonstrated that insulin detemir is a long-lasting preparation similar to what has been observed in humans. A pronounced peak was detected at 8–10 h while the glucose-lowering effect lasted for over 24 h after insulin injection, thus illustrating its longer prolonged peak activity time. Furthermore, intensive glycemic control was achieved with insulin detemir in insulin dependent diabetic dogs, using a lower dosage than NPH insulin and insulin glargine therapeutic doses. Our results indicate that insulin detemir has a greater effect than either NPH insulin or insulin glargine in canines, requiring a lower dose than either insulin preparation. However, using insulin detemir also carries a higher risk of inducing hypoglycemia as compared to either NPH insulin or insulin glargine.     

Feed efficiency and body composition are related to cortisol response to adrenocorticotropin hormone and insulin-induced hypoglycemia in rams

Metabolic rate and energy consumption increase through the activation of the hypothalamic-pituitary-adrenal axis when an animal is exposed to a stressor. Residual feed intake (RFI) as a measure of efficiency has been shown to be related to exogenous adrenocorticotropin hormone (ACTH)-stimulated cortisol concentrations, which is indicative of the relationship between an animal's response to stress and the efficiency with which the energy is used for growth and production. In this study, we tested the hypothesis that sheep with low post-ACTH serum cortisol concentration relative to the other sheep in the flock have lower RFI values and lower cortisol concentrations following insulin-induced hypoglycemia. Adrenocorticotropin hormone (2.0 μg/kg body weight)-stimulated cortisol concentrations were measured in 100 sheep. The extreme responders were selected (n = 12 high cortisol, n = 12 low cortisol), and feed efficiency and body composition parameters were measured. A second ACTH challenge and an insulin challenge were administered. More efficient sheep (more negative RFI value) were found to have lower (P < 0.05) cortisol concentrations following both an ACTH challenge and an insulin challenge. Low-cortisol sheep (low response to ACTH or insulin) were found to have a lower (P < 0.05) proportion of fat tissue in comparison to the high-cortisol animals. These data clearly indicate that an animal's response to exogenous ACTH or insulin-induced hypoglycemia as a stressor is related (P < 0.05) to efficiency of energy use when measured as RFI. These data have important implications in enabling identification of animals that are superior in terms of feed efficiency and for understanding the physiological mechanisms underlying efficiency of energy use.     

Management of hyperkalemia and associated complications in a ferret (Mustela putorius furo) with urinary obstruction requiring surgery

Urethral obstruction is always an emergency for male ferrets and can lead to hyperkalemia. Surgical treatment under general anesthesia can be required. Untreated hyperkalemia before anesthetic induction can be fatal. A 2-year-old, 1.01 kg, neutered male ferret (Mustela putorius furo) was presented for lethargy, hematuria, and stranguria. Radiographs revealed urolithiasis involving the bladder and urethra. When presented for anesthesia, the ferret had hyperkalemia. Before anesthesia, the potassium was normalized rapidly through a combination of saline, dextrose, insulin, and terbutaline. Terbutaline was added to the therapeutic regime as a result of persistent hyperkalemia. Severe hypotension developed during anesthesia which can be partially attributed to the peripheral vasodilatory effect of terbutaline. Hypoglycemia developed as a result of failure to monitor blood glucose level frequently during surgery and suboptimal regime used for insulin and dextrose. Potassium level in a hyperkalemic ferret can be normalized before anesthesia with a combination of therapeutic agents like saline, sodium bicarbonate, insulin, and dextrose. In a case of persistent hyperkalemia, terbutaline can be added to the therapeutic plan. It is important that blood pressure and blood glucose level are monitored closely to prevent hypotension possibly due to terbutaline and hypoglycemia as a result of insulin. The purpose of this report is to describe a case of a ferret with hyperkalemia that needed rapid correction before anesthesia.     

桑叶多糖MLPⅡ对糖尿病模型大鼠肝脏胰岛素抵抗的改善作用

从桑叶中分离纯化获得一种具有降血糖活性的均一多糖组分MLPⅡ,通过检测分析MLPⅡ治疗组糖尿病模型大鼠的胰岛素敏感指数和肝脏生理指标,探究桑叶多糖MLPⅡ对糖尿病模型大鼠肝脏胰岛素抵抗的改善作用。与糖尿病对照组模型大鼠相比,150 mg/kg MLPⅡ治疗组糖尿病模型大鼠肝脏组织中的总胆固醇(TC)、甘油三酯(TG)和丙二醛(MDA)含量分别降低了40.59%、38.58%和52.81%,而肝脏组织中的超氧化物歧化酶(SOD)活性、肝糖原含量和肝脏葡萄糖激酶活性则分别升高了15.88%、198.48%和169.77%,均达到极显著水平。与糖尿病对照组模型大鼠相比,桑叶多糖MLPⅡ治疗组糖尿病模型大鼠的胰岛素敏感指数均明显提高,肝脏的脂肪变性程度明显减轻,其中150 mg/kg MLPⅡ治疗组均达到极显著水平。研究结果提示,桑叶多糖MLPⅡ可显著改善糖尿病模型大鼠的肝脏糖脂代谢和胰岛素抵抗,其作用可能与调节胰岛素的释放和肝脏的氧化应激水平有关。     

Pancreatic insulin-secreting neoplasm (insulinoma) in a West Highland white terrier.

A West Highland white terrier was evaluated because of persistent hypoglycemia and an acute episode of collapse. A pancreatic insulin-secreting neoplasm (insulinoma) was diagnosed on the basis of clinical signs, serum glucose levels, serum insulin levels, abdominal ultrasonography, and exploratory laparotomy with histologic evaluation of neoplastic tissue.     

Toxicological Studies of Phenoxyacetic Herbicides in Animals

The main purpose of the present work was to study the long-term effects of 2,4-dichlorophenoxyacetic acid (2,4-D) in swine, rats and chickens.In preliminary short-term experiments with calves and pigs, definite although reversible toxic effects were seen after single doses of 200 and 100 mg/kg, respectively. Rats and chickens seemed to tolerate 100 and 300 mg/kg, respectively, without ill-effects. On repeated administration daily doses of 50 mg/kg could be toxic to pigs, whereas chickens tolerated 300 mg/kg/day for several weeks without visible effects. Symptoms of acute poisoning in calves were dysphagia, anorexia, tympanites and muscular weakness. Anorexia was apparent also in acutely or subacutely poisoned pigs together with locomotory disturbances, transient diarrhoea and, in severe cases, vomiting, muscular weakness and general depression. In all animals showing symptoms of poisoning a reduced disappearance rate of 2,4-D from plasma was apparent. On autopsy the pigs showed signs of gastro-intestinal irritation and pneumonia and renal degeneration. In the rats and chickens no gross pathological changes were seen.In the long-term studies 5 young pigs were fed 2,4-D (500 p.p.m.) for up to 12 months. Main clinical signs were growth depression, locomotory disturbances, anaemia and albuminuria. Morphological changes included moderate hepatic and renal degeneration. In another experiment 2,4-D was fed to a pregnant sow throughout the gestation period and for 6 further weeks. The sow exhibited no characteristic signs, and on autopsy no changes attributable to 2,4-D were noted. The newborn piglets, however, were underdeveloped and apathetic. Ten out of 15 died within 24 hours. On continued feeding of 2,4-D to the survivors until 7–8 months of age the main effects were a marked growth depression, persistent anaemia and moderate degenerative changes of liver and kidneys.Pregnant rats were given 2,4-D (1000 p.p.m.) in the drinking water during the gestation and further for up to 10 months. The administration of 2,4-D was continued to the second generation rats for up to 2 years. Except for a retarded growth and an increased mortality in the second generation no unequivocal clinical or morphological changes were seen.In chickens continued administration of 2,4-D (500 p.p.m. in the feed or 1000 p.p.m. in the drinking water) caused a reduced eggproduction and pronounced kidney enlargement due to epithelial proliferations, this latter lesion appearing only when very young chicks were used as experimental animals.The experimental results indicate the chronic toxicity of 2,4-D for the species examined to be moderate. Apart from the nephrotoxicity demonstrated in chicks the long-term effects were non-specific. Of particular interest, however, are the high mortality in the newborn piglets and the reduced egg production in the chickens as indications of a possible interference with reproduction.     

Individual heterogeneity in erythrocyte susceptibility to Babesia divergens is a critical factor for the outcome of experimental spleen-intact sheep infections

Susceptibility of sheep erythrocytes to Babesia divergens was investigated in vitro and a high inter-individual variability in their ability to support parasite population development was demonstrated, with some individuals having refractory red blood cells (RBC). As neither changes in growth conditions nor the use of different B. divergens strains influenced the level of susceptibility, the main factor postulated for this variability is the erythrocyte itself. Sheep therefore represent an excellent in vitro model to study the parasite-erythrocyte interaction. In addition, the existence of refractory RBC should help in the identification of the erythrocyte components required for B. divergens development. Experimental infections were carried out on spleen-intact sheep characterized by refractory or fully susceptible erythrocyte types. These differences translated into the successful infection of only those animals with susceptible erythrocytes: infected animals showed no clinical signs, but maintained an asymptomatic persistent infection, as usually observed in the natural bovine host. Sheep therefore represent model organisms that can allow us to study interactions between B. divergens and its vertebrate host at different levels of biological organisation, from the target cell to the intact animal, and represent an experimental infection model of concomitant immunity. Only a low percentage (13%) of the sheep population tested possessed susceptible erythrocytes and the potential role of sheep as a natural host or reservoir of B. divergens is discussed.     

Appropriate use of recovery groups in nonclinical toxicity studies: value in a science-driven case-by-case approach

A recovery phase--a nondosing period that follows the main dosing phase of a study--is sometimes included in nonclinical toxicity studies, and it is designed to understand whether toxicities observed at the end of the dosing phase are partially or completely reversible. For biopharmaceuticals with long half-lives, the inclusion of recovery arms can be helpful in understanding effects of prolonged exposure and assessing antidrug antibodies. This commentary discusses when to include recovery groups in nonclinical toxicity studies, the number of recovery groups to include in a given study, the number of animals to include in each recovery group, and the duration of the recovery phase. In general, the inclusion of recovery arms should follow a case-by-case approach that values rational scientific design and reflects the development needs and regulatory requirements applicable to individual nonclinical programs to ensure appropriate guidance for human studies while minimizing laboratory animal use.     

Effect of inulin and oligofructose enrichment of the diet on rats suffering thiamine deficiency

Thiamine deficiency resulted in inhibition of two main pathways supplying energy to the tissues: glycolysis and β-oxidation. Glycolysis was found to be inhibited to 40% of initial value calculated on the basis of RBC trans-membrane transport of glucose. Prolongation of experiment cause lowering of uptake of this sugar. In rats, energy production from fatty acids (FA) seems to be less sensitive to thiamine deficiency than glycolysis. After 30 days of feeding, utilization of FA in rats was depressed to the 61% of initial value. Thiamine deficiency suppressed insulin secretion, and the changes were statistically significant. Feeding of rats with thiamine restricted diet for 1 month caused the reduction of serum insulin by 14%. In the same animals, trans-membrane glucose transport was reduced over two-times, what might suggest a decreased efficiency of insulin action in such conditions. The kind and concentration of non-digestible fructo-oligosaccharides (FOS) did not affect significantly serum insulin concentration in animals fed thiamine restricted diet. Substitution of a part of wheat starch with FOS has only insignificant compensatory effect on the uptake of glucose. A partial amelioration of the β-oxidation inhibition caused by feeding rats with thiamine deficient diet was observed in animals supplemented with FOS. However, this effect was statistically significant only in rats receiving diet containing 10% of inulin. The effect of supplemented FOS and their concentration on trans-membrane glucose transport in RBC was statistically significant, when pulled supplementation groups were used for statistical evaluation.     

Hepatocellular carcinoma associated with erythrocytosis and hypoglycemia in a yearling filly

A yearling Arabian-type filly with a history of poor growth, erythrocytosis, hypoglycemia, and high liver enzyme activities was admitted to the hospital for evaluation. Three days after admission, the filly collapsed, deteriorated rapidly despite treatment, and was euthanatized. A metastatic hepatocellular carcinoma with capsular rupture and hemoperitoneum were found at necropsy. Primary liver tumors are rare in horses, and hepatocellular carcinoma has been reported in only 1 other horse. The systemic manifestations of the tumor in this filly included weakness, weight loss, inappetence, erythrocytosis with tumor production of erythropoietin, persistent hypoglycemia with normal serum insulin concentrations, serum alpha-fetoprotein (normally present only during fetal life), and terminal massive hemoperitoneum, all features of the syndrome in man.     

Comparison of time-action profiles of insulin Glargine and NPH insulin in normal and diabetic dogs

Intermediate insulin injections are commonly used for glycemic control in insulin dependent diabetic dogs acting as a replacement for natural insulin. Neutral Protamin Hagedorn (NPH) insulin and insulin glargine are two types of injectable insulin preparations commonly used in humans. In our study, we investigated the time-action profiles of both aforementioned insulin preparations in normal dogs in order to determine whether co-administration of NPH and glargine would be of benefit to insulin dependent diabetic dogs as it is for humans suffering from insulin dependent diabetes. Time-action profiles of NPH insulin and insulin glargine in normal dogs demonstrated a clear difference between both insulin preparations confirming that NPH insulin is an intermediate-acting preparation whereas insulin glargine is a long-lasting preparation. In addition, co-administration of NPH insulin and insulin glargine resulted in tight glycemic control as compared to NPH insulin alone in insulin dependent diabetic dogs. However, co-administration result in hypoglycemia at the dosages tested.     

Plasma glucose and cortisol responses to exogenous insulin in fasted donkeys

Susceptibility to equine hyperlipaemia is increased by poor food intake. To assess the contribution of changes in insulin sensitivity, plasma glucose and cortisol responses to an intravenous insulin challenge (0·4 kg⁻¹ bodyweight) were compared with those observed after saline administration in six donkeys fasted either overnight or for three days. Three days of fasting decreased both the rate of insulin-induced hypoglycemia and the maximal hypoglycemic response. A transitory increase in plasma cortisol which peaked within one to four hours of insulin administration was observed in three of the six overnight-fasted donkeys and in all of the three-day fasted donkeys; inter-animal variation appeared to exist in the responsiveness of the hypothalamic-pituitary-adrenocortical ( ) axis to stimulation by insulin-induced hypoglycemia. Fasting is likely to present a risk of equine hyperlipaemia, at least in part, by the reduction in tissue sensitivity to the glucoregulatory action of insulin.     

Obesity induced changes to plasma adiponectin concentration and cholesterol lipoprotein composition profile in cats

Feline obesity generally results in aberrations to plasma metabolite levels, such as lipid concentrations and lipoprotein composition. This study sought to investigate the resultant effect of obesity on cholesterol lipoprotein composition and circulating adiponectin concentrations in cats. Plasma glucose, lipids (triglyceride, cholesterol and free fatty acid), insulin and adiponectin concentrations, and cholesterol lipoprotein composition were measured and compared between body condition score (BCS) determined normal healthy control and obese cats. Although the obese group demonstrated higher levels of plasma cholesterol, glucose, and triglycerides, as compared to healthy controls, the difference was insignificant thus indicating that the BCS determined obese cats may have been overweight and not morbidly obese. Plasma insulin levels were significantly higher (25–30%) versus healthy control animals thereby possibly hinting at the ensuing emergence of obesity induced insulin resistance. However, the BCS determined obese cat demonstrated a significant reduction (p < 0.05) in plasma adiponectin concentration and a significant increase (p < 0.05) in LDL-cholesterol % as compared to age matched healthy control animals. This would indicate that changes in plasma adiponectin concentration and cholesterol lipoprotein composition may be good early indicators of obesity in cats.     

Characteristics of structures and lesions of the eye in laboratory animals used in toxicity studies

Histopathology of the eye is an essential part of ocular toxicity evaluation. There are structural variations of the eye among several laboratory animals commonly used in toxicity studies, and many cases of ocular lesions in these animals are related to anatomical and physiological characteristics of the eye. Since albino rats have no melanin in the eye, findings of the fundus can be observed clearly by ophthalmoscopy. Retinal atrophy is observed as a hyper-reflective lesion in the fundus and is usually observed as degeneration of the retina in histopathology. Albino rats are sensitive to light, and light-induced retinal degeneration is commonly observed because there is no melanin in the eye. Therefore, it is important to differentiate the causes of retinal degeneration because the lesion occurs spontaneously and is induced by several drugs or by lighting. In dogs, the tapetum lucidum, a multilayered reflective tissue of the choroid, is one of unique structures of the eye. Since tapetal cells contain reflecting crystals in which a high level of zinc has been demonstrated chemically, drug-induced tapetum degeneration is possibly related to zinc chelation. The eye of the monkey has a macula similar to that of humans. The macula consists only of cones with a high density, and light falls directly on the macula that plays an important role in visual acuity. Macular degeneration occurring in monkeys resembles histopathologically that of humans. Hence, the eye of the monkey is a suitable model to investigate macular degeneration and to assess drug-induced macular lesions.     

Evaluation of bovine viral diarrhea virus control using a mathematical model of infection dynamics

A mathematical model for infection with bovine viral diarrhea virus (BVDV) was created comprising a series of coupled differential equations. The model architecture is a development of the traditional model framework using susceptible, infectious and removed animals (the SIR model). The model predicts 1.2% persistent infection (within the range of field estimates) and is fairly insensitive to alterations of structure or parameter values. This model allows us to draw important conclusions regarding the control of BVD, particularly with respect to the importance of persistently infected (PI) animals in maintaining BVD as an endemic entity in the herd. Herds without PI animals are likely to experience episodic reproductive losses at intervals of two to three years, unlike herds with PI animals which will not see such marked episodic manifestations of infection. Instead, these herds will experience an initial peak of disease which will settle to low-level chronic reproductive losses. The model indicates that vaccine coverage for herd immunity (to avoid episodic manifestations of disease) need be only 57% without PI animals, although 97% coverage is required when PI animals are present. Analysis of model behavior suggests, a program of detection and removal of PI animals may enhance the effectiveness of a vaccine program provided these animals are in the herd for 10 days or less. The best results would be seen with PI animals in the herd for 5 or fewer days.     

Comparison of a 2-step insulin-response test to conventional insulin-sensitivity testing in horses

Equine insulin resistance is important because of its association with laminitis. The insulin-response test is described to diagnose insulin resistance in clinical settings. Practitioners may be reluctant to perform this test because of the time needed for the test and the fear of inducing hypoglycemia. The objective of the study was to compare a 2-step insulin-response test with a complete insulin-response test. A complete insulin-response test was performed on 6 insulin-resistant horses and 6 controls. A 2-step insulin-response test consisting of an intravenous injection of 0.1 IU/kg human insulin and blood glucose determination at 0 and 30 min after injection was performed on the same horses. Times to reach a 50% reduction of glucose baseline were compared between tests and horses. All the horses tolerated both tests well. No significant difference was observed between baseline glucose concentrations of insulin-resistant horses and controls (P = 0.09). Time to reach 50% reduction of glucose baseline for controls was not significantly different with the use of the complete insulin-response test or the 2-step test (P = 0.98). For insulin-resistant horses, the time to reach 50% reduction of glucose baseline with the use of the 2-step test was significantly longer than for controls (P = 0.004). With a cut-off time of 30 min, the 2-step test had the same characteristics as the complete test. The 2-step test provided a safe, rapid, and low-cost method to diagnose insulin resistance in horses in a clinical setting.