Bovine postparturient hemoglobinemia: hypophosphatemia and metabolic disorder in red blood cells

Mechanism of hemolysis in postparturient hemoglobinemia was studied in 7 cows. Cows 1 to 5 had a history of hemoglobinemia at a previous calving, but hemoglobinemia did not occur during the present parturition. Cow 6, a daughter of cow 4, and cow 7 from another farm, developed postparturient hemoglobinemia and had hemoglobinuria on days 20 and 21 and 10 to 17 after calving, respectively. During the time cows 6 and 7 had hemoglobinuria, both cows had a marked decrease in serum inorganic phosphorus, RBC adenosine 5'-triphosphate (ATP), and reduced glutathione, and a significant (P less than 0.01) increase in methemoglobin concentration. In cow 6, these changes were observed before the onset of hemoglobinuria, indicating metabolic disorder of RBC. After phosphate administration IV, serum inorganic phosphorus was corrected, and RBC ATP was increased above base-line value. During the time cows 6 and 7 had hemoglobinuria, PCV, hemoglobin concentration, and RBC count decreased progressively and reached nadir values, 40% to 50% of baseline values, on day 22 in cow 6 and on day 19 in cow 7. Cows 6 and 7 were anemic, even after serum inorganic phosphorus and RBC ATP values returned to acceptable values. Glycolytic disorder and depletion of ATP, resulting from phosphorus deficiency, appeared to be a primary and essential step leading to hemolysis in postparturient hemoglobinemia in cows 6 and 7.     

Influence of low phosphorus intake during early lactation on apparent digestibility of phosphorus and bone metabolism in dairy cows

In this study, the effects of reducing the dietary phosphorus (P) supply during the first 4 months of lactation on P balance and bone metabolism were investigated in dairy cows during a full lactation, including the dry period. Twenty-two multiparous cows of the Swedish Red and White Breed were included in the experiment. Eleven cows received a dietary P concentration of 0.43% of DM during the whole lactation (NP), and 11 cows received a dietary P concentration of 0.32% of DM during the first 4 months of lactation, followed by 0.43% of DM during the rest of lactation (LP). Total collection of faeces was carried out during five different stages of lactation, for 5 consecutive days at each occasion. The dynamics of bone metabolism was investigated by monitoring one bone-formation marker, identifying osteocalcin (OC), and one bone-resorption marker identifying C-telopeptide fragments of collagen type I (CTx) in blood plasma. At the two first collection periods, 3–7 and 11–15 weeks after parturition, the apparent digestibility of P was higher in the LP cows (52%) than in the NP cows (42%). During the following collection periods, no difference in the apparent digestibility could be noted between the two groups of cows. Phosphorus retention did not differ between treatments at any collection period. The highest retention was observed during late lactation and during the dry period. The profiles of the bone metabolism markers indicated a net resorption of bone during early lactation, but there were no differences in marker concentrations between the groups during early lactation, indicating that the LP diet did not induce a further elevated net bone resorption.     

Assessment of the red blood cell proteome in a dog with unexplained hemolytic anemia

A 7‐year‐old female neutered Jack Russell Terrier was presented to Langford Vets, the University of Bristol, with a history of chronic intermittent lethargy. Investigations and clinical course were compatible with hereditary hemolysis due to a red blood cell membrane defect. Proteomics was used to explore protein alterations in the presence of a hypothesized red blood cell membrane protein deficiency. Proteomic analysis revealed downregulation of the band 3, and alpha‐ and beta‐adducin proteins, and alterations in the red blood cell proteome consistent with previous reports of changes due to the presence of reticulocytosis and ongoing hemolysis. The spectrum of protein alterations identified in the affected dog may be homologous to a band 3 protein deficiency secondary to hereditary spherocytosis, as described in people.     

Genetic markers in the blood and their relation to metabolic parameters in dairy cows

A relationship between the genetic markers of blood (blood groups, serum polymorphic proteins) and the clinico-chemical parameters was studied in the dairy cows of the Slovak Pied breed. Antigens belonging to systems A, B, C, F, S, R, T, Z and the polymorphic traits genetically controlled from loci Tf, Cp, Am and Hb were identified in all the animals subjected to testing. The values of the parameters of acid-base balance and concentration of 13 metabolic components were repeatedly determined in the experimental period. The results of the F-test indicated that there were no significant differences in the values of any of the tested parameters between the phenotypes of the A, J, Am, Tf and Cp systems. Of the 21 parameters tested, statistically significant differences were found in 11 parameters between some alleles of the C, FV, T, Z and Hb systems.     

Biological characteristics for assessing low phosphorus intake in finishing swine

Three replicates of a randomized complete-block design in a 3 x 3 factorial arrangement of treatments involving 141 pigs were conducted to evaluate the effect of low P intake on various response criteria in swine from 65 to 95 kg body weight. Total dietary P levels of .09, .21 and .31% were each fed in a Ca:P ratio of 1.1, 1.5:1 or 2:1 for a 35-d experimental period. The basal diet was formulated using corn starch, dextrin and a soy protein concentrate with monosodium phosphate and limestone supplying desired treatment levels of Ca and P. Blood and bones were collected for mineral analysis. Gain and feed performance responses were not affected by Ca:P ration or dietary P level for the 35-d period. Serum inorganic P concentrations declined as Ca:P ratio increased, but increased as dietary P increased. Serum alkaline phosphatase activity increased with higher Ca:P ratios, and decreased as dietary P level increased. Percent bone ash was not influenced by Ca:P ratio, but increased as dietary P level increased. Bone-bending moment was influenced by Ca:P ratio, and increased linearly as dietary P increased. Dietary P level resulted in a linear increase in total bone and its ash component weight, and in the accretion of the ash and organic matrix component. These data suggest that between 65 to 95 kg body weight, percent bone ash was affected by dietary P level, while bone-bending moment, bone component weights, their accretion, both dietary p level and Ca:P ratio.     

Autoantibodies and red blood cell membrane proteins in a case of canine autoimmune hemolytic anemia.

Autoantibodies and erythrocyte membrane proteins were analyzed in a case of a dog with autoimmune hemolytic anemia (AIHA). In crisis phase, antiglobulin test was positive. The eluate from the erythrocytes of the dog with AIHA gave agglutination against autologus erythrocytes. Immunoglobulin subclasses in the eluate were revealed to be IgG and IgA by the double diffusion test. Comparing the SDS polyacrylamide gel electrophoretic patterns of erythrocyte membrane proteins between the crisis and remission phases, there was a change in the protein on protein 4.1 region. However, there were no changes in blood group typings in two phases.     

Biological characteristics for assessing low phosphorus intake in growing swine

A total of 195 swine from 18 to 35 kg were used to evaluate various biological responses of growing swine fed low P diets at different Ca:P ratios. Three replicates of a randomized complete-block design in a 3 X 3 factorial arrangement of treatments were used. Total dietary P levels of .12, .31 and .50% were fed in 1:1, 1.5:1 or 3:1 Ca:P ratios. Diets were formulated using corn starch, dextrin and soy protein concentrate, with monosodium phosphate and limestone used to supply appropriate dietary treatment levels of Ca and P. Gain and feed performance were generally influenced by both Ca:P ratio and dietary P level. Serum inorganic P decreased linearly as Ca:P ratio widened and increased linearly as dietary P increased. Serum Ca concentrations responded inversely to serum inorganic P in response to both Ca:P ratio and dietary P level. Serum alkaline phosphatase was not affected by Ca:P ratio, but declined quadratically with increasing dietary P level. Bone bending moment was influenced by both dietary P level and Ca:P ratio. Bone component weights (ash, organic matrix), percentage bone ash and net accretion of bone ash and organic component weights were not influenced by Ca:P ratio but increased linearly as dietary P level increased. These results suggest that bone component weights, net ash accretion and percentage bone ash were more sensitive criteria than bone bending moment or serum alkaline phosphatase, which in turn were more sensitive than serum inorganic P, Ca, Mg and growth performance characteristics in response to dietary P level.     

牧草磷素营养及其耐低磷特性

磷是牧草重要的营养元素之一, 土壤的磷肥力往往成为牧草正常生长发育的限制因子。本文综述了磷肥对牧草生长的影响, 比较了磷肥与其他营养元素对牧草的综合影响, 阐述了牧草对低磷胁迫的适应性, 介绍了接种真菌和培育磷高效牧草品种可提高牧草对低磷环境的适应力, 指出利用分子生物学技术开展磷高效牧草育种和种质资源的筛选是解决我国牧草有效利用磷素问题的关键。     

Mortality attributable to bluetongue virus serotype 8 infection in Dutch dairy cows

In 2007, bluetongue virus serotype 8 (BTV-8) re-emerged in the Netherlands and a large number of farmers notified morbidity and mortality associated with BTV-8 to the authorities. All dead cows in the Netherlands are registered in one of the three age classes: newborn calves <3 days, calves 3 days to 1 year, and cows >1 year. These registrations result in a complete data set of dead cattle per herd per day from 2003 until 2007. In this study, the mortality associated with BTV-8 for the Dutch dairy industry was estimated, based on this census data. Default, mortality associated with BTV-8 was estimated for the confirmed notification herds. Moreover, an additional analysis was performed to determine if mortality associated with BTV-8 infection occurred in non-notification herds located in BTV-8 infected compartments. A multivariable population-averaged model with a log link function was used for analyses. Separate analyses were conducted for the three different age groups. Confirmed notification herds had an increased cow mortality rate ratio (MRR) (1.4 (95% CI: 1.2-1.6)); calf MRR (1.3 (95% CI: 1.1-1.4)); and newborn calf MRR (1.2 (95% CI: 1.1-1.3)). Furthermore, in non-notification herds in BTV-8 infected compartments, mortality significantly increased 1.1 times (95% CI: 1.1-1.1) in cows, 1.2 times (95% CI: 1.2-1.2) in calves and 1.1 times (1.1-1.1) in newborn calves compared with BTV-8 non-infected months. Using objective census data over a 5-year period, the MRRs indicated increased mortality associated with BTV-8 infection not only in herds of which the farmer notified clinical signs but also in non-notification herds in infected compartments.     

低磷血红蛋白尿症奶牛红细胞膜溶破的分子机理

利用自然病例 ,采取分组 (血尿组、低磷组、对照组 )对比的方法 ,从分子水平上检测了红细胞膜的磷脂组分、膜骨架蛋白组分及红细胞形态扫描电镜观察。结果表明 ,血尿组奶牛红细胞膜的磷脂组分和膜骨架蛋白及红细胞形态均发生明显地改变 :血尿组奶牛红细胞膜磷脂酰胆碱 磷脂酰丝氨酸 (PC PS)及神经鞘磷脂 (SM)显著高于低磷组和对照组 ,而磷脂酰乙醇胺 (PE)含量显著低于低磷组和对照组 ,血清磷与红细胞膜 PE含量呈显著正相关 ,与 SM呈显著负相关 ;血尿组奶牛红细胞膜膜收缩蛋白 、 及区带 - 2蛋白含量均显著低于低磷组和对照组 (P<0 .0 1)而区带 蛋白含量明显高于低磷组和对照组(P<0 .0 1) ;从扫描电镜下观察发现 ,由低磷发展至溶血 ,红细胞形态从正常的圆盘状→棘形→球形→溶破的演变过程 ,发生血管内溶血时 ,其红细胞膜磷脂组分、骨架蛋白及红细胞形态发生显著改变 ,是导致溶血的最关键性因素     

Controversies related to red blood cell transfusion in critically ill patients

Objective – To review the evolution of and controversies associated with allogenic blood transfusion in critically ill patients. Data sources – Veterinary and human literature review. Human Data Synthesis – RBC transfusion practices for ICU patients have come under scrutiny in the last 2 decades. Human trials have demonstrated relative tolerance to severe, euvolemic anemia and a significant outcome advantage following implementation of more restricted transfusion therapy. Investigators question the ability of RBCs stored longer than 2 weeks to improve tissue oxygenation, and theorize that both age and proinflammatory or immunomodulating effects of transfused cells may limit efficacy and contribute to increased patient morbidity and mortality. Also controversial is the ability of pre‐ and post‐storage leukoreduction of RBCs to mitigate adverse transfusion‐related events. Veterinary Data Synthesis – While there are several studies evaluating the transfusion trigger, the RBC storage lesion and transfusion‐related immunomodulation in experimental animal models, there is little research pertaining to clinical veterinary patients. Conclusions – RBC transfusion is unequivocally indicated for treatment of anemic hypoxia. However, critical hemoglobin or Hct below which all critically ill patients require transfusion has not been established and there are inherent risks associated with allogenic blood transfusion. Clinical trials designed to evaluate the effects of RBC age and leukoreduction on veterinary patient outcome are warranted. Implementation of evidence‐based transfusion guidelines and consideration of alternatives to allogenic blood transfusion are advisable.     

Osmotic stress in red blood cells from beagles with hemolytic anemia.

Red blood cell populations separated by density centrifugation were compared in a dynamic assay of osmotic stress. Red blood cells from Beagles genotypically normal and nonanemic (nonaffected), Beagles with inherited hemolytic anemia (anemic), and Beagles presumed to be carriers of the anemia trait (trait carriers) were examined for rate and extent of swelling after exposure to the ionophore A23187 in a medium containing calcium and potassium chloride. Comparisons were made between RBC populations separated on the basis of density. Significant differences were observed in the rates of cell swelling in RBC populations separated by density between nonaffected and anemic Beagles. The response of RBC from Beagles presumed to carry the anemia trait was similar to that of RBC from nonaffected dogs. One phenotypic expression of this inherited abnormality of RBC in Beagles was an accelerated rate of RBC swelling under osmotic stress, and this swelling response diminished with increasing RBC density.     

Assessment of magnesium intake according to requirement in dairy cows

To date, no specific hormonal regulation system has been identified for homoeostatic control of the essential mineral Mg. In cattle, the maintenance of physiological plasma Mg concentration depends on gastrointestinal absorption, primarily from the rumen, which serves as a pool for covering the requirement. Whereas a possible surplus (absorption greater than requirement) is rapidly excreted by the kidneys, a shortage (absorption lower than requirement) cannot be compensated for by mobilization from the large Mg pool in bones or soft tissue, so that the maintenance of the necessary physiological Mg concentration in plasma relies on continuous and sufficient absorption. Our knowledge concerning the site and mechanisms of Mg absorption has improved during the last few decades, and meta‐analyses of the absorption of Mg in dairy cows have shown that the K content has a pronounced negative effect on Mg digestibility. The current recommendations of Mg intake propose a constant percentage of Mg and emphasize the depressive effect of high potassium (K) intake on Mg absorption. The current knowledge about the antagonism between K intake and Mg absorption allows a more flexible solution which includes the K content of the diet. An assessment of Mg intake is proposed that incorporates the improved knowledge of Mg absorption, metabolism and requirement. Within this framework, an equation is derived that allows a prediction of the amount of Mg required to compensate for dietary K content, the goal being to avoid both possible undernutrition or an unnecessary surplus of dietary Mg.     

Natural antibodies related to metabolic and mammary health in dairy cows

Natural antibodies (NAb) are defined as antibodies that circulate in normal healthy individuals under the absence of deliberate antigenic stimulation. Two types of NAb are distinguished: NAb towards exogenous antigens and NAb towards autoantigens (N(A)Ab). The objectives of the current study were threefold. First, we studied the relation between metabolic health and concentrations of NAb binding keyhole limpet hemocyanin (KLH) or lipopolysaccharide (LPS) in milk and plasma of dairy cows in early lactation. Second, we determined the presence of N(A)Ab binding transferrin, myosin and thyroglobulin in bovine milk. Third, we studied the relation between N(A)Ab in bovine milk and mammary health. For the first objective, dairy cows were either fed a control (C) (n = 8) or a diet where 2 kg of concentrates were replaced by an iso-energetic concentrate containing marine algae (ALG) from week −3 till 8 postpartum (experiment 1). Plasma and milk samples were analyzed weekly for NAb binding either KLH or LPS. Plasma was analyzed for glucose, non-esterified fatty acids (NEFA) and β-hydroxybutyrate (BHBA). For the second and third objective, milk samples were collected weekly from 96 dairy cows from week 2 till 9 postpartum and analyzed for milk composition and N(A)Ab binding myosin, transferrin and thyroglobulin (experiment 2). For both datasets, N(A)Ab titers are expressed as ²log values of the highest dilution giving a positive reaction. Data are expressed as means ± SEM. Repeated observations were analyzed in a mixed model. In experiment 1, no diet effect (P > 0.05) was observed on NAb binding LPS in plasma or milk, NAb binding KLH in milk was greater (P = 0.05) for cows fed the control diet. Concentration of NAb binding KLH and LPS in plasma was negatively related to plasma NEFA concentration (P < 0.05). In experiment 2, NAb binding myosin (5.66 ± 0.06), thyroglobulin (4.85 ± 0.06), and transferrin (5.76 ± 0.07) were identified in milk. Clinical mastitis incidence (9%) tended to be positively related to concentration of NAb binding myosin (P = 0.06) and negatively related to Nab binding transferrin (P = 0.08). In conclusion, NAb binding KLH and LPS in plasma and milk are related to metabolic health, as indicated by plasma NEFA concentration. Furthermore, this study demonstrates the presence of N(A)Ab in bovine milk and shows trends for a relation between N(A)Ab binding auto-antigens and mastitis. Future studies should confirm these trends and shed light on the predictive value of N(A)Ab in bovine milk for mammary health.     

Macrohematuria and life-threatening anemia attributable to subepithelial vascular ectasia of the urinary bladder in a dog

Hematuria leading to life-threatening anemia was evaluated in a 9-year-old male dog. At surgery, an abnormal vessel resembling an ulcerated varicose vein was found on the surface of the bladder mucosa. After removal of the lesion, hematuria ceased and had not recurred 22 months later.