Erythrocyte osmotic fragility testing and the prediction of canine malignant hyperthermia susceptibility

A Doberman-German Shepherd cross-bred male dog, previously diagnosed as malignant hyperthermia susceptible, was mated to an unrelated nonsusceptible German Shepherd cross-bred female. The resultant litter was subjected to hematological, biochemical and erythrocyte osmotic fragility testing in an endeavor to predict the susceptibility of individuals to malignant hyperthermia. Laboratory evaluations were repeated at one year of age and the litter subjected to the halothane challenge test. No significant difference in erythrocyte osmotic fragility was found between malignant hyperthermia susceptible and nonsusceptible siblings at six weeks or at one year of age. Erythrocyte osmotic fragility, in both malignant hyperthermia susceptible and nonsusceptible animals, increased between six weeks and one year of age. Dantrolene sodium was an effective treatment for malignant hyperthermia in the dog when administered early in an episode and in adequate dosage. The initial sign of a malignant hyperthermia episode was a very rapid increase in end tidal partial pressure of carbon dioxide. This finding reinforces the value of capnographic monitoring in anesthesia.     

The use of the rapid osmotic fragility test as an additional test to diagnose canine immune-mediated haemolytic anaemia

Background

Diagnosing canine immune-mediated haemolytic anaemia (IMHA) is often challenging because all currently available tests have their limitations. Dogs with IMHA often have an increased erythrocyte osmotic fragility (OF), a characteristic that is sometimes used in the diagnosis of IMHA. Since the classic osmotic fragility test (COFT) is time-consuming and requires specialized equipment, an easy and less labour-intensive rapid osmotic fragility test (ROFT) has been used in some countries, but its diagnostic value has not yet been investigated.This study aimed to evaluate erythrocyte osmotic fragility in dogs with and without IMHA, to compare results of the classic (COFT) and rapid (ROFT) test and to assess the value of the ROFT as diagnostic test for canine IMHA.Nineteen dogs with IMHA (group 1a), 21 anaemic dogs without IMHA (group 1b), 8 dogs with microcytosis (group 2), 13 hyperlipemic dogs (group 3), 10 dogs with lymphoma (group 4), 8 dogs with an infection (group 5) and 13 healthy dogs (group 6) were included.In all dogs, blood smear examination, in-saline auto-agglutination test, Coombs’ test, COFT and ROFT were performed. In the COFT, OF5, OF50 and OF90 were defined as the NaCl concentrations at which respectively 5, 50 and 90% of erythrocytes were haemolysed.

Results

Compared with healthy dogs, OF5 and OF50 were significantly higher in group 1a (P < 0.001) and OF5 was significantly higher in group 3 (P = 0.0266). The ROFT was positive in 17 dogs with IMHA, 10 hyperlipemic dogs, one anaemic dog without IMHA and one healthy dog.

Conclusions

Osmotic fragility was increased in the majority of dogs with IMHA and in dogs with hyperlipidemia, but not in dogs with microcytosis, lymphoma or an infection. Although more detailed information was obtained about the osmotic fragility by using the COFT, the COFT and ROFT gave similar results. The ROFT does not require specialized equipment, is rapid and easy to perform and can be used easily in daily practice. Although, the ROFT cannot replace other diagnostic tests, it may be a valuable additional tool to diagnose canine IMHA.     

Training-induced modifications in some biochemical defences against free radicals in equine erythrocytes

Oxidative stress develops when the generation of free radicals exceeds the antioxidant capacity of cells or extracellular fluids. It can also occur as a result of physical exercise, and the pathogenesis of exercise-induced myopathies and haemolysis in horses may be related to changes in lipid peroxidation caused by free radicals. Cells have developed biochemical protection against oxidative stress and, as tissues seem to increase their antioxidant defences under chronic activation, training may be one of the ways of increasing antioxidant defences. Accordingly, we tested some enzymatic antioxidant activities as well as nonenzymatic antioxidants in horses undergoing special training. The results indicated a decrease in both chemical and biochemical defences against free radicals during training. It was deduced that the horses' diet may have been unable to provide the increased need for antioxidant defences resulting from training.Abbreviations DPH 1,6-diphenyl-1,3,5-hexatriene - EU enzymic unit - GSH-Px glutathione peroxidase - P degree of fluorescence polarization - Phospholipid P phosphorous content in phospholipids - RBC red blood cell - t-ButOOH t-butylhydroperoxide     

Effect of Dictyocaulus filaria infection on the osmotic fragility of sheep erythrocytes

The osmotic fragility of erythrocytes in lambs experimentally infected with Dictyocaulus filaria was studied weekly for 71 weeks. In acute infection, the erythrocytic fragility increased from the third week of infection onwards, reached its peak by the eleventh week and declined thereafter. However, in the chronic immune-carrier stage, this increase in the fragility did not return to normal until the end of the experiment. This enhanced fragility showed a positive correlation with the faecal larval count, worm burden and the extent of lung damage in lambs.     

The effect of Eperythrozoon suis infection on the osmotic fragility of erythrocytes]

Osmotic fragility of erythrocytes was tested in weaned pigs experimentally infected with Eperythrozoon (E.) suis. Acute eperythrozoonosis of splenectomized pigs led to an increase of osmotic fragility. It is supposed that E. suis infection causes a structural change in erythrocyte membrane. Possible mechanisms of this cell membrane injury are discussed.     

Erythrocyte osmotic fragility and general health status of adolescent Sprague Dawley rats supplemented with Hibiscus sabdariffa aqueous calyx extracts as neonates followed by a high‐fructose diet post‐weaning

High‐fructose diets (HFD) can cause oxidative damage to tissues including erythrocyte cell membranes. Hibiscus sabdariffa (HS) has protective antioxidant properties. Rats were used to investigate whether the consumption of HS by neonates would result in long‐term effects on their erythrocyte osmotic fragility (EOF) and general health when later fed a high‐fructose diet post‐weaning through adolescence. Eighty of four‐day‐old Sprague Dawley rat pups were divided randomly into three treatment groups. The controls (n = 27) received distilled water at 10 ml/kg b. w, while the other groups received either 50 mg/kg (n = 28) or 500 mg/kg (n = 25) of an HS aqueous calyx extract orally till post‐natal day 14. The rats in each group were weaned and divided into two subgroups; one continued on normal rat chow, and the other received fructose (20% w/v) in their drinking water for 30 days. Blood was collected in heparinised tubes and added to serially diluted (0.0–0.85%) phosphate‐buffered saline to determine the EOF. Clinical markers of health status were determined with an automated chemical analyser. HS extracts did not programme metabolism in the growing rats to alter their general health and EOF in response to the HFD.     

铜胁迫对山羊红细胞渗透脆性和溶血度的影响

通过体外培养红细胞加入不同浓度硫酸铜溶液,采用改良的渗透法测定红细胞的渗透脆性和酸化甘油试验测定其溶血度。结果显示,铜显著增加山羊红细胞渗透脆性（MCF）并呈现明显的时间剂量效应;铜引起红细胞半数溶血度（AGLT50）明显降低,且呈明显的时间剂量效应。过量铜能引起红细胞渗透脆性增加,红细胞溶血度增加,使红细胞损伤加重。     

Anemia, splenomegaly, and increased osmotic fragility of erythrocytes in Abyssinian and Somali cats

OBJECTIVE: To determine clinical and clinicopathologic features of a chronic intermittent severe hemolytic anemia characterized by erythrocyte osmotic fragility in Abyssinian and Somali cats. DESIGN: Case series. ANIMALS: 13 Abyssinian and 5 Somali cats. PROCEDURES: History, pedigree information, and results of routine laboratory tests, special erythrocyte studies, and histologic evaluation of splenic and hepatic specimens were analyzed. RESULTS: Age at which clinical signs of anemia were first apparent ranged from 6 months to 5 years. Ten cats had splenomegaly. Most often, the PCV was between 15 and 25%, but it was as low as 5% at some times. The anemia was characterized by macrocytosis and mild to moderate reticulocytosis, but no poikilocytosis. Hyperglobulinemia, lymphocytosis, mild hyperbilirubinemia, and high hepatic enzyme activities were common findings. Results of Coombs tests and tests for infectious diseases were negative. The erythrocytic osmotic fragility was high in affected cats (mean osmotic fragility, 0.66 to 0.78%), compared with healthy cats (0.48 to 0.58). No specific membrane protein abnormality, erythrocyte enzyme deficiency, or hemoglobinopathy was identified. Histologic evaluation of splenic and hepatic specimens revealed extramedullary hematopoiesis and hemosiderosis. Four of the 5 Somali cats were closely related. CONCLUSIONS AND CLINICAL RELEVANCE: On the basis of results of pedigree analyses, the apparent breed predilection, and the exclusion of other known causes of anemia in cats, we believe that the hemolytic anemia in these cats was likely a result of a novel hereditary erythrocyte defect. A genetic predisposition to immune-mediated destruction of erythrocytes could not be ruled out.     

Effect of ascorbic acid administration on erythrocyte osmotic fragility of pigs transported by road during the hot-dry season

The experiments were performed in order to determine the effect of 8-h road transportation of pigs on erythrocytes osmotic fragility during the hot-dry season, and the ameliorative role of ascorbic acid. Twenty-three adult pigs comprising of both sexes served as subjects for the study. Thirteen pigs administered ascorbic acid (AA) per os 30 min before transportation, at a dose rate of 100 mg/kg served as experimental animals, while ten pigs administered with distilled water per os served as control, and were transported for 8 h during the hot-dry season. EDTA blood samples collected a day before (pre-transportation), immediately after 8-h transportation and 7 days post-transportation were used to determine erythrocyte osmotic fragility. The ambient temperature (AT) and relative humidity (RH) measured within the vehicle ranged between 30.5 – 39.0°C and 40.0 – 71.0% respectively. These values were outside the thermoneutral zone for the pig, indicating that the season was thermally stressful. Results obtained showed a significant difference (p < 0.05) in percent haemolysis recorded at NaCl concentrations of 0.4% and 0.6% immediately after transportation in experimental pigs and at 0.5, 0.6, 0.8 and 0.9% NaCl concentrations in experimental pigs 7 days post-transportation. In conclusion, result from the present study indicated that 8-h road transportation during the hot-dry season could induce stress resulting in haemolysis of erythrocytes and AA administration ameliorated the stress.     

铜缺乏对乳牛自由基代谢的影响

为阐明铜缺乏对乳牛体内自由基代谢的影响,本实验选择缺铜地区全血铜低于NRC下限(0.8mg/kg)但无明显摇摆症状的犊牛25头作为亚临床铜缺乏组;并对其中15头进行补铜试验(饲料中添加硫酸铜100mg/kg干重),以15头临床健康乳牛作为对照组,分别于40天及80天采集样并检测全血中自由基的含量及血清中NO(一氧化氮)的含量;同时检测了发病乳牛与对照乳牛(各9头)组织细胞浆中NO的含量。结果表明:发病组及亚临床组乳牛全血自由基含量均显著高于对照组(P〈0.05),各组乳牛血清中NO含量差异均显著(P〈0.05),且发病乳牛大脑、小脑及脊髓NO的含量显著或极显著高于对照组,补铜能够显著降低亚临床缺乏组乳牛全血中的自由基或血清中NO含量。结论:铜缺乏能够影响响乳牛体内的自由基代谢过程。     

Evaluation of the Schirmer tear test in clinically normal rabbits

Mean (+/- SD) value for results of the Schirmer tear test without use of topical anesthesia on 142 rabbit eyes was 5.30 (+/- 2.96) mm/min; range was 15 mm/min, with low value of 0 and high value of 15 mm/min. Two SD from the mean provided a normal clinical range of 0 to 11.2 mm/min for inclusion of 95% of the population. Significant differences were not evident between eye (right vs left) or gender or among breeds; interaction among eye, gender, and breed also was not observed. Although mean values for 2 breeds were significantly different from values in the other 10 breeds and from each other, they were still within the proposed normal range of 2 SD. The Schirmer tear test without topical anesthesia may be useful in rabbits for evaluation of increased values correlated with ocular irritation, rather than for determination of decreased values associated with keratoconjunctivitis sicca.     

NO自由基在镉致育成鸡卵巢损伤中的作用

近年来,随着镉消费的增加和消费形式的改变,全国每年由工业废弃物排放到环境中的镉总量约680余吨,镉污染区覆盖畜禽约10亿头(只)、人口近1亿[1].镉对生殖系统的毒性较为敏感,已受到国内外的普遍关注,并进行了一系列的动物实验和人群流行病学研究[2-3].但有关镉对禽类生殖功能,尤其是对雌性生殖系统的影响,目前尚未见报道,本试验以健康育成鸡为实验动物,通过观察卵巢形态结构变化,检测血清和卵巢中NO含量及NOS活性,探讨镉对禽类雌性生殖毒理机制,为防治禽类镉中毒提供试验依据.     

Erythrocyte selenium-75 uptake as a measure of selenium status in weaner sheep, and its relationship to erythrocyte glutathione peroxidase activity

The relationship between in vitro erythrocyte 75Se uptake (75Se uptake) and erythrocyte glutathione peroxidase (EGSHPx) activity was examined in weaner sheep during periods of selenium depletion and repletion, to determine whether 75Se uptake was better correlated than EGSHPx activity to the development of weaner nutritional myopathy. In the 2 trials conducted, only 3 of 45 Merino wether weaners developed clinical myopathy and histological lesions in skeletal muscles. The 75Se uptake values and EGSHPx activities in these 3 sheep were no different from those in the unaffected sheep. There was a significant negative correlation between 75Se uptake values and EGSHPx activities over the entire period of the trials. It could not be demonstrated that 75Se uptake was any better correlated than EGSHPx activity to the development of nutritional myopathy, and it was concluded that EGSHPx activity indicated selenium status better than 75Se uptake in weaner sheep.     

Effect of different kinds of anoxia/reoxygenation on the mitochondrial function and the free radicals production of cultured primary equine skeletal myoblasts

Horses are outstanding athletes, performing in many different disciplines involving different kinds of efforts and metabolic responses. Depending on exercise intensity, their skeletal muscle oxygenation decreases, and the reperfusion at cessation of the exercise can cause excessive production of free radicals. This study on cultured primary equine myoblasts investigated the effect of different kinds of anoxia/reoxygenation (A/R) on routine respiration, mitochondrial complex I specific activity and free radicals production. Our data revealed that short cycles of A/R caused a decrease of all the parameters, opposite to what a single long period of anoxia did. A preconditioning-like effect could explain our first pattern of results whereas mild uncoupling could be more appropriate for the second one. Anyway, it seems that mitochondrial complex I could play a major role in the regulation of the balance between metabolic and antioxidant protection of the muscular function of athletic horses.