Beta-globin chain hemoglobin polymorphism and hemoglobin stability in black rhinoceroses (Diceros bicornis)

To evaluate the syndrome of acute intravascular hemolytic anemia in black rhinoceroses (Diceros bicornis), the hemoglobin of this species was evaluated by use of isopropanol- and heat-stability tests and was further characterized by electrophoretic studies. Samples were obtained from 22 apparently healthy captive North American black rhinoceroses, though 3 of the study animals had survived previous hemolytic events, and 3 others were parents of 3 offspring that had suffered hemolysis. The eastern African (Diceros bicornis michaeli) and the southern African subspecies (D b minor) were represented. Comparative samples were also obtained from 2 white (Ceratotherium simum) and 1 Indian (Rhinoceros unicornis) rhinoceroses. The hemoglobin of all 3 species appeared stable when tested by use of the heat and isopropanol methods. Thus, an unstable hemoglobin does not appear to be involved in the hemolytic crises of captive black rhinoceroses. Black rhinoceros hemoglobin had a striking polymorphism. Thirteen of the samples from black rhinoceroses had a single hemoglobin band, based on results of alkaline electrophoresis. Nine had, in addition to this major band, a slow (more cathodic) minor band that comprised about 10% of the total hemoglobin. Further studies indicated that the major band and the slower minor band may contain globin chains analogous to human beta- and delta-chains respectively; these bands have been tentatively designated B and C. Phenotypes B and BC are common, in a ratio of 4:3. A genetic mechanism is proposed that assumes beta b and beta c gene loci and that beta c-locus-expressed (beta c+) and beta c-locus-inhibited (beta c degrees) are common alleles for the beta c-locus.(ABSTRACT TRUNCATED AT 250 WORDS)     

Effect of hemoglobin polymorphism on performance traits in indigenous chicken genotypes in Nigeria

Tropical Animal Health and Production - The effect of hemoglobin polymorphism on performance traits in Nigerian indigenous chicken types was investigated in this research. The chickens were...     

Genetic polymorphism of bovine beta-casein gene in Japanese dairy farm herds

The aim of this study was to investigate beta-casein polymorphism among 320 Japanese cows sampled from eight dairy farms. We used a newly-developed genotyping method that involved collecting DNA from hairs and a Cycleave polymerase chain reaction (PCR) assay to detect the A1, A2, and B variants. Results revealed the presence of five genotypes (A1A1, A2A2, A1A2, A1B, and A2B). We found that the most common genotype was A2A2 (0.42), followed by A1A2 (0.39) and A1A1 (0.11). The A1B and A2B genotypes were less frequent (<0.05). The frequencies of alleles A1, A2, and B were calculated to be 0.32, 0.64, and 0.04, respectively. Our study is the first to show the current status of beta-casein polymorphisms in Japanese dairy farms. Given the adverse effects of A1 beta-casein on human health, attempts have been made to develop herds consisting solely of A2A2 cows. Our study provides a reference for improving cow populations in Japanese dairy farms. The Cycleave PCR-based assay we developed here can be used for rapid and reliable genotyping of bovine beta-casein.     

牛体外受精胚的发育速度和染色体研究

利用不同个体的牛冷冻精液对屠宰场废弃牛卵巢内卵母细胞进行体外受精,对体外受精胚胎的发育速度和染色体的异常发生进行了研究。结果显示,冷冻精液A组体外受精48h后的卵裂率显著高于冷冻精液B组(P<0.05),2组之间的囊胚形成率差异显著(P<0.05);冷冻精液A体外受精后的胚胎发育速度显著快于冷冻精液B组(P<0.05);染色体分析结果表明,2组胚胎染色体异常发生率无显著差异,异常胚胎均表现为多倍体的发生率较高。     

Prevalence of coagulase gene polymorphism in Staphylococcus aureus isolates causing bovine mastitis.

This study was conducted to investigate polymorphism of the coagulase gene of Staphylococcus aureus causing bovine mastitis. One hundred eighty-seven strains of S. aureus were isolated from bovine mastitic milk samples obtained from 187 different Danish dairy farms. The isolates were characterised for restriction fragment length polymorphism (RFLP) of the coagulase gene. A variable region of the coagulase gene was amplified using the polymerase chain reaction (PCR) followed by AluI restriction enzyme digestion. A total of 15 different RFLP patterns were observed. The predominant pattern was found in 35% of the isolates. The ease of analysing coagulase gene polymorphisms among a large number of strains, and the multiple distinct polymorphic patterns generated, supports the use of this technique in epidemiological investigations of bovine mastitis. The predominating variants may have predelection for causing intramammary infections.     

Developmental competence of frozen-thawed blastocysts from fair-quality bovine embryos cultured with beta-mercaptoethanol

Two-hundred-and-thirty-one fair-quality embryos at the compacted morula stage collected from 89 superovulated cows were cultured in TCM199 or Brinster's BMOC-3 medium with or without 100 microM beta-mercaptoethanol (beta-ME). After 24 h culture, a total of 142 fair-quality embryos developed to the blastocyst stage, of which 106 were subsequently frozen with 1.8 M ethylene glycol. The mean cell number and development rates of frozen-thawed blastocysts from the fair-quality embryos cultured in TCM199 containing beta-ME were higher than those of the fair-quality embryos directly frozen without culture. The pregnancy rates obtained with frozen blastocysts from fair-quality embryos tended to be lower than those of non-cultured fresh fair-quality embryos and cultured fresh blastocysts. These results indicate that the inclusion of beta-ME in pre-freezing culture media improve the development of frozen-thawed blastocysts from fair-quality embryos, but not the pregnancy rate.     

奶牛乳腺乳脂肪形成相关基因的遗传多态性

牛奶营养品质低下已成为我国奶业健康发展所面临的严峻挑战。乳脂肪是决定牛奶营养品质的主要物质之一。调控乳腺内脂肪酸转运、从头合成、酯化和脂滴形成的相关基因的遗传多态性对乳脂肪的成分及含量具有重要影响。本文综述了这些基因的遗传多态性及其对乳脂肪生成的影响,旨在为明确奶牛乳腺脂肪酸形成调控网络提供理论基础和研究思路。     

奶牛乳房炎金黄色葡萄球菌基因多态性分型试验

利用随机引物AP-7,建立引物随机多态性扩增(RAPD)体系对71株引起内蒙古和贵州地区奶牛乳房炎的金黄色葡萄球菌分离株进行基因分型研究。结果表明,71株金黄色葡萄球菌均得到清晰的RAPD指纹图谱,扩增产物为2～9条带,产物大小为240～4 500bp。菌株共分为6个基因型,其中Ⅰ型17株(占23.9%)、Ⅱ型3株(占4.2%)、Ⅲ型33株(占46.5%)、Ⅳ型15株(占21.1%)、Ⅴ型2株(占2.8%)、Ⅵ型2株(占2.8%)。Ⅰ型为内蒙古地区的流行优势菌群,Ⅲ型为贵州地区的流行优势菌群。两地区各基因型菌株比例有明显差异,这可能与奶牛养殖业水平和环境差异有关。     

Polymerized bovine hemoglobin (Oxyglobin Solution) administration in two river otters (Lutra canadensis)

Polymerized bovine hemoglobin (Oxyglobin Solution^®) was successfully administered to two river otters (Lutra canadensis) that required general anesthesia and surgery for trap‐related injuries. In both animals, blood oxygen content was maintained at presurgical levels despite a 47–70% decrease in their hematocrit. Otter 1 received a dose of 19 mL kg^?1, given at a rate of 29 mL kg^?1 h^?1. Otter 2 received a dose of 20 mL kg^?1, given at a rate of 5 mL kg^?1 h^?1. For many of the uncommon species, there are limited resources in the form of blood products to treat anemia, blood loss or hypovolemia. Successful use of polymerized bovine hemoglobin in these two otters suggests that it is useful for acute treatment of anemia and blood loss in such species.     

Analysis of methanogens in the bovine rumen by polymerase chain reaction single-strand conformation polymorphism

The polymerase chain reaction single‐strand conformation polymorphism (PCR‐SSCP) method reported by Schwieger and Tebbe (1998) was used to analyze the diversity of methanogens inhabiting the rumen. Partial 16S rRNA gene fragments were amplified from DNA extracted from rumen contents by PCR with archaea‐specific primers, Ar1000F and Ar1500R, or methanogen‐specific primers, M301F and M915R, with one primer phosphorylated at the 5′ end. The amplified DNA fragments were analyzed by SSCP gel electrophoresis after the phosphorylated strands of the PCR products were digested with λ exonuclease. When we analyzed samples collected from the six Holstein cows used in a previous study, in which cows were given feed with or without α‐cyclodextrin‐horseradish oil complex (CD‐HR), nine and six bands were identified in the profiles generated by PCR products amplified with archaea‐specific and methanogen‐specific primers, respectively. While dendrogram analysis based on SSCP gel profiles found that the methanogens from each rumen showed a particular composition of methanogens, the profiles of the methanogens isolated from two of three cows fed with CD‐HR fell into the same branch in the dendrogram constructed from the profiles. Therefore, this study demonstrates the potential of the PCR‐SSCP method in the methanogenic community analysis of the rumen and in investigating changes in the methanogenic community due to the addition of CD‐HR to the rumen.     

Use of bovine single nucleotide polymorphism markers to verify sample tracking in beef processing

OBJECTIVE: To determine whether a selected set of 20 single nucleotide polymorphism (SNP) markers derived from beef cattle populations can be used to verify sample tracking in a commercial slaughter facility that processes primarily market (ie, culled) dairy cows. DESIGN: Prospective, blinded validation study. ANIMALS: 165 cows and 3 bulls from 18 states (82% Holstein, 8% other dairy breeds, and 10% beef breeds). PROCEDURE: Blood was collected by venipuncture from randomly chosen animals just prior to slaughter. The purported corresponding liver samples were collected during beef processing, and genotype profiles were obtained for each sample. RESULTS: On the basis of SNP allele frequencies in these cattle, the mean probability that 2 randomly selected individuals would possess identical genotypes at all 20 loci was 4.3 x 10(-8). Thus, the chance of a coincidental genotype match between 2 animals was 1 in 23 million. Genotype profiles confirmed appropriate matching for 152 of the 168 (90.5%) purported blood-liver sample pairs and revealed mismatching for 16 (9.5%) pairs. For the 16 mismatched sample pairs, 33% to 76% of the 20 SNP genotypes did not match (mean, 52%). Discordance that could be attributed to genotyping error was estimated to be < 1% on the basis of results for split samples. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that this selected set of 20 bovine SNP markers is sufficiently informative to verify accuracy of sample tracking in slaughter plants that process beef or dairy cattle. These or similar SNP markers may facilitate high-throughput, DNA-based, traceback programs designed to detect drug residues in tissues, control of animal diseases, and enhance food safety.     

Inadequacy of low-volume resuscitation with hemoglobin-based oxygen carrier hemoglobin glutamer-200 (bovine) in canine hypovolemia

Stroma-free hemoglobin-based oxygen carriers (HBOC) have been developed to overcome problems associated with transfusion of allogeneic blood. We have studied the efficacy of the first licensed veterinary blood substitute, hemoglobin glutamer-200 bovine (Oxyglobin; Biopure, Cambridge, MA, USA, Hb-200), in a canine model of acute hypovolemia and examined whether clinically commonly used criteria are adequate to guide fluid resuscitation with this product. Twelve anesthetized dogs were instrumented for measurements of physiological variables including hemodynamic, oxygenation, and blood gas and acid-base parameters. Dogs were bled to a mean arterial pressure (MAP) of 50 mmHg for 1 h followed by resuscitation with either shed blood (controls) or Hb-200 until heart rate (HR), MAP and central venous pressure (CVP) returned to baseline. Recordings were repeated immediately and 3 h after termination of fluid resuscitation. Hemorrhage (average 32 mL/kg) caused significant decreases in total hemoglobin (Hb), mean pulmonary arterial pressure (PAP), cardiac output (CO) and oxygen delivery (DO2I), increases in HR and systemic vascular resistance (SVRI), and lactic acidosis. In controls, only re-transfusion of all shed blood returned HR, MAP and CVP to prehemorrhage values, whereas in other dogs this endpoint was reached with infusion of 10 mL/kg Hb-200. Unlike blood transfusion, Hb-200 infusion failed to return CI and DO2I to baseline and to increase arterial oxygen content (CaO2) and total Hb; SVRI further increased. Thus, commonly used criteria (HR, MAP, CVP) to guide transfusion therapy in patients posthemorrhage prove insufficient when HBOCs with pronounced vasoconstrictive action are used and lead to inadequate volume repletion.     

Effects of a polymerized ultrapurified bovine hemoglobin blood substitute administered to ponies with normovolemic anemia

The development of ultrapurified hemoglobin-based oxygen carriers has eliminated many problems associated with whole-blood transfusions in other species. We hypothesized that the administration of polymerized ultrapurified bovine hemoglobin (PUBH) would result in improved hemodynamic parameters in ponies with normovolemic anemia without adverse effects on renal function or coagulation times. Normovolemic anemia was induced in 6 healthy adult ponies. Over a 3-day period, at least 45 mL/kg of whole blood was withdrawn from each pony until a target PCV of <12% was attained. Plasma was separated from the red blood cells via centrifugation and readministered to the ponies on each day. After the final plasma transfusion, 15 mL/kg of hetastarch (control, n = 6) or 15 mL/kg of PUBH (treatment, n = 6) was administered at 10 mL/kg/h IV. Administration of PUBH at a rate of 10 mL/kg/h was not associated with any adverse effects in 5 of the 6 ponies. One pony experienced an anaphylactoid reaction during infusion of PUBH. The reaction, characterized by intense pruritus, tachycardia, and tachypnea resolved shortly after stopping the infusion. Ponies receiving PUBH had significantly lower cardiac indices (P = .03) and heart rates (P = .002) than control animals. A significantly greater increase in central venous pressure was observed in the PUBH group compared to the hetastarch group (P = .02). No adverse renal or coagulation effects were observed with PUBH infusion. These results suggest that PUBH improves hemodynamics and oxygen transport parameters in horses experiencing normovolemic anemia. Patients should be monitored closely during infusion for any adverse reactions.     

Phylogenetic relationships of Staphylococcus aureus from bovine mastitis based on coagulase gene polymorphism

Coagulase gene restriction fragment length polymorphism (RFLP) patterns were analyzed to determine the phylogenetic relationship among isolates of Staphylococcus aureus from the Czech Republic (n = 27), France (n = 48), Korea (n = 115) and the United States (n = 278). A total of 468 isolates of S. aureus were subtyped into 41 coagulase genotypes. Cluster analysis placed the 41 types into nine clusters. Eighteen API Staph profiles were determined for 102 S. aureus isolates representing 1 to 4 isolates of each coagulase type. The results of the study suggest that based on coagulase gene RFLP analysis, several genetic variants of S. aureus are prevalent. Comparison of coagulase and API Staph profiles indicated that the two identification system were independent of each other.     

Prion protein gene polymorphism and blood lymphocyte profile in cows naturally infected with bovine leukemia virus

The polymorphic loci of the bovine prion protein (PRNP) gene, comprising 23-bp insertion/deletion (23-bp indel) within the promoter sequence and 12-bp insertion/deletion (12-bp indel) within the intron 1 sequence, are located in regions which play a key role in gene expression. The objective of this study was to determine whether the 23-bp and 12-bp insertion/deletion polymorphism within the PRNP gene leads to significant differences in the blood lymphocyte profile and to investigate changes in the composition of these cells in cattle naturally infected with Bovine Leukemia Virus. An analysis of the effect of the bovine PRNP gene polymorphism on the blood lymphocyte profile revealed considerable differences between animals with the 23-bp indel genotypes, and small and statistically non-significant differences between those with the 12-bp indel genotypes. 23-bp del/del homozygotes had a significantly lower percentage of T lymphocytes with the phenotypes CD2 (P < 0.01), CD8 (P < 0.01) and WC1-N2 (P < 0.05), and a higher ratio of CD4 to CD8 T lymphocytes, compared to animals with the 23-bp ins/ins genotype. The obtained results indicate that the 23-bp indel polymorphism, in contrast to the 12-bp indel polymorphism, has a significant effect on changes in the blood lymphocyte profile. The size of blood lymphocyte subpopulations was also found to change under the influence of enzootic bovine leukosis. The direction of those changes in EBL-positive animals is consistent with that observed in 23-bp del/del homozygotes, which may testify to the adverse effect of this genotype on immunological efficiency.