Haemoglobin types of old Norwegian short tail land race sheep

Thirty years have passed since it was shown that sheep have two types of haemoglobin (Hb), A and B. Since then a large number of studies have been undertaken; gene frequencies, relations and origins of breeds, biochemistry, correlations with diseases, to mention the most important fields for which these 2 haemoglobins have been utilized. A few years ago, in a number of studies, Vestri and coworkers (Vestri et al. 1983) using chromatographic and biochemical methods together with isoelectric focusing showed that in Italian sheep additional Hb types occurred. These new types are caused by substitutions in the alpha chains, in contrast to the A and B difference which is caused by 7 amino acid substitutions in the beta chain (Huisman et al. 1965). Vestri et al (1983) also proved that the alpha chains in sheep, as in man, horse and goat among others, are controlled by 2 closely linked alpha globin genes.     

Haemoglobin oxygen affinity and regulating factors of the blood oxygen transport in canine and feline blood

Complete dynamic oxygen equilibrium curves (OEC) on dogs and cats whole blood were measured at 33, 37 and 41 degrees C. OEC were also run at three partial carbon dioxide pressures (20, 40 and 80 mmHg) as well as at five pH levels (7.2, 7.3, 7.4, 7.5 and 7.6). 2,3- diphosphoglycerate (DPG) concentrations were determined. Results were compared to those previously published in humans, using the same experimental method [Comp. Biochem. Physiol. 106 (1993) 687]. In standard conditions (pH 7.4, pCO2 40 mmHg and temperature 37 degrees C), the partial oxygen pressure at half-saturation of haemoglobin (p50) was 30.0+/-1.3 mmHg for dogs and 34.1+/-1.8 mmHg for cats. Cat's OEC was thus rightshifted compared to dog's OEC, itself rightshifted compared to human OEC. 2,3-DPG concentrations were higher in dogs than in men until they were very low in cats. Contrary to that observed in human medicine, no significant correlation was identified between standard p50 and canine 2,3-DPG values. Influence of pH, pCO2 and temperature on the OEC was saturation dependent. In dogs, Delta log p50/Delta pH was equal to -0.370, Delta log p50/Delta log pCO2 was 0.093 and Delta log p50/Delta T was 0.020. In cats, Delta log p50/Delta pH was equal to -0.405, Delta log p50/Delta log pCO2 was 0.080 and Delta log p50/Delta T was 0.016. Practically, temperature and pH variations exert a lesser influence in domestic carnivores than in humans, effect of pCO2 being similar in both.     

Analysis of relationships between German heavy horse breeds based on pedigree information

We analysed the relationship coefficients (R) between the four German heavy horse breeds South German Coldblood, Rhenish German Draught Horse, Schleswig Draught Horse and Black Forest Draught Horse. The relationship coefficient makes it possible to ascertain crossbreeding between the breeds over time, or autonomous developments of the breeds, respectively. The investigation revealed that the relationship coefficients between the German draught horse breeds were very low. The mean relationship coefficients between the four German heavy horse breeds were largest between the South German Coldblood and Schleswig Draught Horse (0.103%), whereas mean relationship coefficients were lowest between the Rhenish German and Black Forest Draught Horse (0.001%). The Rhenish German Draught Horse showed largest relationship coefficients with the Schleswig Draught Horse (0.09%), while the Black Forest Draught Horse was mostly related to the South German Coldblood (0.06%). The results reveal the presence of very few common progenitors of the breeds. The gene flow between the breeds is primarily due to crossbreeding of stallions and, especially, in the Rhenish German Draught Horse population breeding with a few mares from other German draught horse breeds.     

Amyloid in the horse: a report of nine cases

Out of approximately 16,000 horses referred for clinical examination, nine had amyloidosis. Six of these horses had localised amyloid deposits in the wall of the nasal meatus and ventral turbinates associated with epistaxis. Horse 1 also developed malignant histiolymphocytic lymphosarcomas. The amyloid deposits were potassium permanganate-resistant and tryptophan-positive. Gel filtration of solubilised amyloid fibrils from Horse 1 revealed a major retarded fraction with an apparent molecular weight of 20 kD. This protein had an amino acid composition similar to human AL-amyloid proteins and horse immunoglobulin light chains. On Western blot a strong cross-reaction was observed between horse 1gG2a light chains and the Horse 1 amyloid. Horses 7 to 9 had suppurative verminous aneurysm, tuberculosis and an adrenal cortical adenoma, respectively, and had generalised amyloid deposits in liver and spleen. These amyloid deposits were found to be potassium permanganate-sensitive and positive for tryptophan. Gel filtration of solubilised amyloid fibrils from Horse 8 revealed a major retarded fraction (protein AA) with an apparent molecular weight of 10 kD. Immunoperoxidase-antiperoxidase staining showed the localised deposits to be negative or only weakly positive with antisera against bovine, hamster, dog and human protein AA and to be positive with anti-horse-one amyloid protein. The generalised deposits were found to be positive with the antisera against allogenic protein AA. The results of the potassium permanganate incubation, biochemistry, immunoblotting and immunochemistry, indicate that the localised amyloid of Horse 1 and most likely the amyloid of Horses 2 to 6, is of the AL-type. The generalised amyloid deposits were found to be of the AA type.     

Oxygen dissociation curves of whole blood from the Egyptian free-tailed bat,Tadarida aegyptiaca E. Geoffroy,using a thin-layer optical cell

Tadarida aegyptiaca (mean body mass 13.5 g) is a fast flying insectivorous bat that hunts in open areas for extended periods, covering extensive distances during its foraging bouts. Whole blood samples taken from the wing arteries were analysed for 2,3-diphosphoglyceric acid, oxygen affinity and pH. The mean oxygen affinity of T. aegyptiaca blood with a P₅₀-value 38.3 mm Hg (5.1 kPa) is lower than that of similar-sized bats but not as low as the P₅₀-values of 48.5 mm Hg (6.4 kPa) in two mice species. This indicates that oxygen affinity of the blood in T. aegyptiaca and other bats is designed to facilitate both effective binding of oxygen with haemoglobin at the alveolar/blood capillary interface and oxygen offloading in the tissues. Haematocrits (54–60 %) and levels of 2,3 DPG (2.36–3.30 mM l^–1) were higher, and blood pH (7.106–7.229) lower, than those reported for terrestrial species of a similar size, but within the range of values reported for other bats. We conclude that adequate delivery of oxygen to tissues with high metabolic demands during flight is facilitated primarily by an elevated haematocrit and a reduced blood pH in response to elevated levels of metabolic by-products such as 2,3-DPG (from the citrate acid cycle) and CO₂ (leading to increased concentrations of H⁺ ions).     

An in vitro evaluation of storage media for the preservation of canine packed red blood cells

The effect of four different red blood cell storage media on in vitro parameters of stored canine red blood cells was studied. The storage media included citrate-phosphate-dextrose-adenine (CPDA-1), two additive solutions, and an additive solution modified by the addition of plasma. Biochemical and hematologic parameters, including red cell adenosine triphosphate (ATP); 2,3-diphosphoglycerate (2,3-DPG); pH; percent hemolysis; and supernatant sodium, potassium, and glucose were assessed immediately following preparation of the red cell concentrate and after 35 and 42 days of storage at 4 degrees C. All parameters changed significantly (p < 0.05) during storage. Significant differences due to effect of the storage media were also seen at each time period. After 35 days and 42 days of storage, CPDA-1 maintained the highest pH, potassium, and sodium values, and had the lowest 2,3-DPG, ATP (p=0.052), and glucose values. No differences were seen in hemolysis after 35 days of storage. No additional benefit was noted from the addition of plasma to the additive solution. The additive solutions compared favorably with CPDA-1.     

Acquisition of haemoglobin-bound iron by Histophilus somni

Histophilus somni is an important pathogen of cattle and sheep. H. somni requires iron and can use ruminant transferrins as iron sources for growth. Here, we investigated the abilities of bovine (strains 649 and 2,336) and ovine (strains 9L and 3384Y) isolates of H. somni to acquire iron from haemoglobins. Using growth assays, the bovine isolates were shown to acquire iron from bovine haemoglobin, but not from ovine, porcine or human haemoglobins; the ovine isolates, however, failed to use any of these haemoglobins as iron sources for growth. In solid phase binding assays, the bovine isolates, grown under iron-restricted conditions in the presence of bovine haemoglobin, bound not only bovine but also ovine and human haemoglobins. Competition binding assays indicated that all three haemoglobins were bound by the same receptor(s) and SDS-PAGE of membrane fractions revealed that expression of haemoglobin-binding activity was associated with the production of an approximately 120-kDa outer membrane protein. PCR approaches allowed the amplification and sequencing of hgbA, and also hugX and hugZ homologues from strains 649, 9L and 3384Y. While hgbA of strain 649 was predicted to encode an HgbA precursor that is processed to yield a mature, 123.9-kDa haemoglobin-binding protein, the hgbA genes of strains 9L and 3384Y were predicted to give rise to truncated products. RT-PCR experiments revealed that in strain 649, hugX, hugZ and hgbA are co-transcribed and iron-regulated and additional sequencing suggested that in strain 2336, expression of HgbA is subject to phase variation involving a poly C tract within hgbA.     

Calculation of bovine haemoglobin oxygen saturation by algorithms integrating age,haemoglobin content,blood pH,partial pressures of oxygen and carbon dioxide in the blood,and temperature

In human and veterinary medicine, arterial and venous haemoglobin oxygen saturations are often used to estimate the severity of a disease and to guide therapeutic decisions. In veterinary medicine, haemoglobin oxygen saturation (SO(2)) is usually calculated using a blood gas analyser and algorithms developed for humans. It is possible, therefore, that the values obtained in animals may be distorted, particularly in animals with a high haemoglobin oxygen affinity, like young calves. In order to verify this hypothesis, we compared the arterial (SaO(2)) and venous (SvO(2)) haemoglobin oxygen saturations calculated using three different algorithms, and the oxygen exchange fraction (OEF) at the tissue level, which is the degree of haemoglobin desaturation between arterial and venous blood (SaO(2)-SvO(2)), with the values obtained from the whole bovine oxygen equilibrium curve (OEC) determined by a reference method. The blood gas analysers underestimated SvO(2) values; consequently, the OEF was overestimated (by about 10%). Two methods of reducing these errors were assessed. As the haemoglobin oxygen affinity decreases during the first month of life in calves a relationship between PO(2) at 50% haemoglobin saturation (P50) and age was established in order to correct the calculated values of venous and arterial SO(2), taking into account the estimated position of the OEC. This method markedly reduced the error for SvO(2) and OEF. Secondly, the SO(2) was calculated using a mathematical model taking into account the age of the animal and the specific effects of pH, PCO(2), and temperature on the bovine OEC. Using this method, the mean difference between the OEF values calculated using the mathematical model and those calculated by the reference method was close to zero. The errors produced by blood gas analysers can thus be minimised in two ways: firstly, by simply introducing a P50 estimated from the age of the calf into the analyser before the measurement; and secondly, by calculating the SO(2) using a mathematical model applied to the bovine OEC.     

Research Progress on Prevalence of Horse Gastric Myiasis

Horse gastric myiasis is a chronic wasting parasitosis caused by larvae of Gasterophilus spp. parasitics on the gastrointestinal tract of the equids. Horse gastric myiasis leads to host highly anaemic, angular, poisoning even death. This disease is distributes all over the world, each country has a slightly different epidemic law, because of different geographical position and climatic environment, and whether using insect repellent or not. Our country is mainly popular in the Northeast, Northwest and Inner Mongolia, which of 6 kinds of popular horse Gastrophilus, and the dominant species are G. intestinalis and G. nasalis. In this paper,the author described the infections of horse gastric myiasis at home and abroad in recent years, summarized epidemic law and put forward the research direction, so as to provide reference for the disease prevention and control.     

马胃蝇蛆病流行病学研究进展

马胃蝇蛆病是由马胃蝇幼虫寄生于马属动物胃肠道内所引起的一种慢性消耗性寄生虫病,导致宿主高度贫血、消瘦、中毒,严重时衰竭死亡。该病呈世界性分布,不同国家因地理位置、气候环境不同及是否广泛使用驱虫药,呈现出不同的流行规律。在中国主要流行于东北、西北及内蒙古等地,流行6种马胃蝇,以肠胃蝇、红尾胃蝇为优势虫种。文章主要对近些年国内外马胃蝇蛆病感染情况进行描述,总结流行规律,提出研究方向,以期为该病的防制提供参考。     

Comparison of 4 blood storage methods in a protocol for equine pre-operative autologous donation

OBJECTIVE: To compare viability of equine whole blood stored by 4 different methods, and to establish optimal storage protocols for an equine autologous blood donation program. STUDY DESIGN: In vitro study of stored equine whole blood. Animals- Six healthy adult horses. METHODS: Blood from each horse was collected into 4 different containers: glass bottles containing acid-citrate-dextrose solution (ACD), plastic bags containing ACD, citrate-phosphate-dextrose (CPD), and CPD with supplemental adenine (CPDA-1). Blood was stored for 5 weeks and sampled at 2-day intervals. Standard hematologic and biochemical variables were evaluated, and adenosine-5-triphosphate (ATP) and 2,3-diphosphoglycerate (2,3-DPG) concentrations were measured and normalized to total hemoglobin content. RESULTS: Plasma hemoglobin, % hemolysis, lactate, potassium, ammonia, and lactate dehydrogenase (LDH) increased, whereas glucose concentration and pH decreased in all stored blood over 5 weeks. There was a temporal increase in hemolysis with all storage methods, but the increase was greatest in glass bottles. Lactate and ammonia were highest in CPD and CPDA-1 samples, indicating more active red blood cell (RBC) metabolism. 2,3-DPG concentrations decreased during storage, but were optimally preserved with CPDA-1. ATP concentrations were significantly higher for blood stored in CPDA-1, and were lowest in glass bottles. CONCLUSIONS: Hematologic and biochemical values measured for blood stored in CPDA-1 are suggestive of improved RBC viability compared with other storage methods. With the exception of ATP, results from stored equine blood were similar to those reported for other species. CLINICAL RELEVANCE: Commercial CPDA-1 bags appear to be the optimal storage method for equine whole blood.     

中国鸭源H5N6 AIV血凝素和神经氨酸酶基因特征及遗传进化分析

为明确中国鸭源H5N6 AIV的HA和NA基因特征及其遗传变异情况,研究选取了NCBI流感病毒资源库里的32株具有完整ORF编码区的中国鸭源H5N6 AIV的HA和NA基因序列,利用分子生物学软件分析其HA和NA基因特征及其遗传进化情况。结果显示,中国鸭源H5N6 AIV的HA裂解位点有REKRRKR↓G、RERRRKR↓G和KEKRRKR↓G三种类型,HA有6个潜在的N-糖基化位点,其中第182、222、224位氨基酸依次为N、Q、G,具有与禽源唾液酸α-2,3受体结合的特性,然而HA发生了S123P/T、T156A、S223R氨基酸位点突变,部分毒株发生了I151T氨基酸位点突变;HA基因核苷酸遗传进化分析显示,中国鸭源H5N6 AIV遗传进化上均处于clade2.3.4.4分支,并进一步进化出Ⅰ和Ⅱ两个亚分支。部分中国鸭源H5N6 AIV NA第59~69位缺失11个氨基酸,中国鸭源H5N6 AIV NA基因遗传进化上分为两个大的分支,分别对应NA基因颈部缺失型和完整型两种类型;NA颈部完整型的NA有6个潜在的N-糖基化位点,NA颈部缺失型的NA由于第59~69位氨基酸的缺失,导致其第67位丢失了一个N-糖基化位点。研究为中国鸭源H5N6 AIV的生物学特性和遗传进化特征研究奠定基础。     

Evaluating the Technique of Using Nitrogen Retention as a Response Criterion for Amino Acid Studies in the Horse

Six Quarter Horse yearling fillies were used in a duplicated 3 × 3 Latin square experiment to evaluate the technique of using nitrogen retention as a response criterion for amino acid studies in the horse. Yearlings were paired by age and randomly assigned to one of three concentrates fed with medium-quality Coastal Bermudagrass hay. Diet A, a soybean meal–based concentrate, was amino acid sufficient. Diet B, a cottonseed hull–based concentrate containing 2% urea, was amino acid deficient. Horses fed diet C, identical to diet B, were orally dosed with synthetic essential amino acids and cysteine, immediately before eating, to match amino acid levels in diet A. Nitrogen retention was not different between diets A and B. There was a trend for horses fed diet C to have higher nitrogen retention, as a result of higher nitrogen intake. Nitrogen retained as a percent absorbed was lower (P < .05) for diet B than for diet A, whereas values for diet C were not different from diets A or B. Nitrogen retention was not an effective response criterion in this study. Differences in nitrogen retained as a percentage absorbed were observed between amino acid–sufficient and deficient diets. However, horses fed amino acid–deficient diets and dosed with synthetic amino acids likely require some modified dosage level to achieve the same or higher values as those for amino acid–sufficient diets.     

Effects of dietary horse beans (Vicia faba) on colostrum and milk composition and milk yield in sows

In an experiment sows received 0, 17 and 34% horse beans in the diet. Horse beans replaced the protein mixture (soybean meal, fishmeal, and meat and bone meal) by one half or in full.The composition of colostrum was altered by the dietary horse beans. An increase in dietary horse beans significantly reduced dry matter contents (P ≤ 0.05) and protein contents (P ≤ 0.001). As to the various protein fractions, total whey protein and albumin showed the greatest decline (P ≤ 0.001), followed by immunoglobulin (P ≤ 0.01) and beta lactoglobulins (P ≤ 0.05). Casein and alpha lactalbumin contents were not altered significantly.The total fat content of colostrum was not influenced by dietary horse beans, but the percentages of C 18:1 increased (P ≤ 0.001) and C 18:2 decreased (P ≤ 0.001) with increasing amounts of horse beans in the diet.The effect of horse beans on the composition of sow milk was insignificant, although there was a tendency to higher dry matter and protein content in milk from sows fed diets without horse beans.Horse beans at both levels had a negative effect on milk yield (linear trend: P ≤ 0.05).     

Bilateral arytenoid cartilage paralysis after inhalation anesthesia in a horse

An 8-year-old Quarter Horse gelding was anesthetized for surgical exploration and debridement of a chronic draining wound in the intermandibular space. Anesthesia was without complication other than persistently low PaO2. Severe airway obstruction was evident immediately after extubation, requiring tracheostomy. Endoscopic diagnosis was bilateral arytenoid paralysis, which gradually resolved over the next 7 days. Compression, trauma, or tension of the recurrent laryngeal nerves are the postulated causes of idiopathic laryngeal hemiplegia in horses. The extremely extended position of the head and neck during anesthesia, perhaps compounded by low arterial oxygen content, may have resulted in a hypoxemic insult to the recurrent laryngeal nerves and caused bilateral arytenoid paralysis in this horse.     

Evaluation of Citrate-Phosphate- Dextrose-Adenine as a Storage Medium for Packed Canine Erythrocytes

Packed canine red blood cells (RBCs) stored in the anticoagulant-preservative solution citrate-phosphate-dextrose-adenine (CPDA-1) were studied at 1, 10, 20, 30, and 40 days. The extracellular concentrations of potassium and sodium, erythrocyte mean corpuscular volume, and osmotic fragility increased during storage (P less than 0.05). There was a decrease in the pH, plasma concentration of glucose, and erythrocyte concentrations of 2,3-diphosphoglycerate (2,3-DPG) and adenosine-5'-triphosphate (P less than 0.05). Erythrocyte 2,3-DPG concentration decreased by 54% within the first 24 hours of storage (P less than 0.001). Posttransfusion viability (PTV) decreased from 90% on day 1 to 46% on day 40 (P less than 0.05). The PTV of the RBCs stored for 10 and 20 days complied with the Food and Drug Administration (FDA) standard. Although there are marked biochemical and hematologic changes in stored packed red blood cells (pRBCs), 20-day-old units may be expected to be of acceptable quality. The sharp decrease in 2,3-DPG concentration suggests a reduction in oxygen carrying capacity in erythrocytes stored as pRBCs. Hyperkalemia occurs during storage of pRBCs and does not appear to be associated with high intraerythrocytic potassium concentrations.     

Heart rate variability after horse trekking in leading and following horses

Horse trekking (HT) is having a stroll on a horse along a walking trail in a forest, field, and/or sandy beach. Generally in HT, horses exercise in tandem line outside the riding facilities. Because the leading horse will be confronted with stressors in the forefront, we hypothesized that the leading horse shows higher stress responses than the following one. In order to verify the hypothesis, we compared short‐term stress responses between each position in six horses. Exercise consisted of 15 min of ground riding and 45 min of HT with walking and trotting. Heart rate variability was analyzed for 5 min at 30, 60, and 90 min after the exercising period. There was no significant difference in heart rate during exercise between leading and following positions. The high frequency / low frequency power band of heart rate variability, an index of sympathetic nervous activity, after exercise, tended to be higher in the leading position than following one (P < 0.1). The result in this study can suggest that the leading horse was in a higher stressed state than the following horse after HT.     

Characterization of the cDNA and genomic DNA sequence encoding for the platelet integrin alpha IIB and beta III in a horse with Glanzmann thrombasthenia

Glanzmann thrombasthenia (GT) is characterized by a defect of platelet aggregation. This autosomal recessive genetic disorder is caused by an abnormality of the platelet glycoprotein receptors alpha IIb or beta III. Recently, we identified a horse with clinical and pathological features of GT. The aim of this study was to describe this case of GT at the molecular level. A point mutation from G to C in exon 2 of ITGA2B causing a substitution of the expected amino acid arginine 72 (Arg(72)) by a proline (Pro(72)) was encountered. This amino acid change may result in abnormal structural conformations that yield an inactive alpha IIb subunit. The genomic DNA analysis showed that this horse was homozygous for the missense mutation.     

Osteochondrosis-like lesion of the anconeal process in two horses

A suspected lesion of osteochondrosis dissecans involving the anconeal process of the humeroradial joint (elbow) was found in 2 horses. In horse 1, the lesion was found during routine lameness examination, during which alleviation of the lameness was achieved after intra-articular anesthesia of the elbow. In horse 2, the lesion was found at necropsy. Horse 1 responded satisfactorily to intra-articular treatment with hyaluronic acid. Both horses also had other joints affected with osteochondrosis dissecans.