The primary structure of the hemoglobin of Malayan sun bear (Helarctos malayanus, Carnivora) and structural comparison to other hemoglobin sequences

The complete primary structure of the alpha- and beta-chains of the hemoglobin of Malayan Sun Bear (Helarctos malayanus) is presented. After cleavage of the heme-protein link and chain separation by RP-HPLC, amino-acid sequences were determined by Edman degradation in liquid- and gas-phase sequenators. An interesting result of this work is the demonstration that the hemoglobin of Malayan Sun Bear is identical to the hemoglobins of Polar Bear (Ursus maritimus) and Asiatic Black Bear (Ursus tibetanus). The paper gives an updated table of identical hemoglobin chains from different species. This paper may be considered as a compilation of work on the genetic relationship of Pandas.     

Comparative Analysis and Molecular Characterization of a Gene BANF1 Encoded a DNA-Binding Protein During Mitosis from the Giant Panda and Black Bear

Barrier to autointegration factor 1 (BANF1) is a DNA-binding protein found in the nucleus and cytoplasm of eukaryotic cells that functions to establish nuclear architecture during mitosis. The cDNA and the genomic sequence of BANF1 were cloned from the Giant Panda (Ailuropoda melanoleuca) and Black Bear (Ursus thibetanus mupinensis) using RT-PCR technology and Touchdown-PCR, respectively. The cDNA of the BANF1 cloned from Giant Panda and Black Bear is 297 bp in size, containing an open reading frame of 270 bp encoding 89 amino acids. The length of the genomic sequence from Giant Panda is 521 bp, from Black Bear is 536 bp, which were found both to possess 2 exons. Alignment analysis indicated that the nucleotide sequence and the deduced amino acid sequence are highly conserved to some mammalian species studied. Topology prediction showed there is one Protein kinase C phosphorylation site, one Casein kinase II phosphorylation site, one Tyrosine kinase phosphorylation site, one N-myristoylation site, and one Amidation site in the BANF1 protein of the Giant Panda, and there is one Protein kinase C phosphorylation site, one Tyrosine kinase phosphorylation site, one N-myristoylation site, and one Amidation site in the BANF1 protein of the Black Bear. The BANF1 gene can be readily expressed in E. coli. Results showed that the protein BANF1 fusion with the N-terminally His-tagged form gave rise to the accumulation of an expected 14 kD polypeptide that formed inclusion bodies. The expression products obtained could be used to purify the proteins and study their function further.     

Primary structure of hemoglobin of the polar bear (Ursus maritimus, Carnivora) and the Asiatic black bear (Ursus tibetanus, Carnivora

The adult Polar and Asiatic Black Bear have one hemoglobin component each. The complete amino-acid sequences of their alpha- and beta-chains are presented. Their primary structures were determined by sequencing the tryptic and prolyl peptides. The alignment of these peptides was deduced from homology to human hemoglobin chains. The hemoglobin sequences of the two species proved to be identical. The evolutionary aspects of this result are discussed. A table of identical hemoglobin sequences from different species is given.     

Comparative endocrine investigations in three bear species based on urinary steroid metabolites and volatiles

In order to improve breeding of in situ populations of bears, a comprehensive study of reproductive physiology in Brown (Ursus arctos), Spectacled (Tremarctos ornatus) and Giant panda bears (Ailuropoda melanoleuca) was performed. The objective was to perform non-invasive analyses of urinary and fecal steroid metabolites. In addition, we investigated the presence of reproduction-related urinary volatile substances of these bears that might trigger the reproductive behavior. Urinary estrogen concentrations, routinely used to monitor follicular activity in Giant panda, were inappropriate for monitoring follicular activity in Spectacled bear. In addition, no estrogen peak related to mating activity was observed in Brown bear. Further contrasting Giant panda, although urinary pregnanediol analyses failed to indicate luteal activity in either Spectacled or Brown bears, urinary (Spectacled bear) and fecal (Brown bear) concentrations of progesterone were an appropriate indicator of luteal activity. The Giant panda had volatile components (medium-chain fatty acids) in their urine that increased simultaneously with the seasonal increase of estrogens. These fatty acids were also detected in the Brown during estrus and Spectacled bear. Further studies on the behavioral relevance of these fatty acids are required to determine if they are pheromones.     

Carnivora: the primary structure of the beach marten (Martes foina, Mustelidae) hemoglobin

The primary structures of alpha- and beta-chains from the hemoglobin of the Beach Marten (Martes foina, Carnivora) are presented. The globin chains were separated on CM-cellulose in 8M urea buffer. The amino-acid sequences were established by automatic liquid- and gas-phase Edman degradation of the intact chains and the tryptic peptides from oxidized chains. Comparison of the sequences with human hemoglobin shows 21 exchanges in the alpha- and 12 in the beta-chains. The differences concerning heme and interchain contact sites as well as the substitution alpha 77 (EF6)Pro----Ala are discussed. The latter is observed for the first time in a mammalian hemoglobin. The sequences are compared with those of other Carnivora. The beta-chains of Martes foina and Pteronura brasiliensis (Giant Otter) are found to be identical, but their alpha-chains differ in 7 positions. The surprising small numbers of exchanges between the hemoglobin from Beach marten and that from Lesser and Greater Panda are discussed.     

佛坪自然保护区食竹鸟兽种类的初步调查

1991~2004年采用检查死亡个体的胃或嗉囊内容物、新鲜粪便和在野外直接观察动物采食的3种方法,在陕西佛坪国家级自然保护区对食竹的鸟兽种类进行了初步调查。巴山木竹和秦岭箭竹是大熊猫采食的主要食物资源。除大熊猫外,采食这两种竹的鸟兽有38种,隶属6目14科,其中鸟类9种,兽类29种。大量采食、中等采食和少量采食竹类的物种分别为1种、5种和33种,所占比例分别为2.6%、12.8%和84.6%。39种鸟兽均采食竹笋,其中32种只采食竹笋。黑熊和野猪是食竹笋量比较大的大型兽类,羚牛、鬣羚和斑羚3种牛科动物全年都采食竹类,其中以羚牛对竹叶的采食量最大。这些食竹鸟兽一方面直接与大熊猫竞争食物资源,特别是竹笋,另外也会间接地影响大熊猫对竹类的利用。     

长白山自然保护区黑熊和棕熊种群数量动态分析

1986～2010年冬季采用路线调查方法,对吉林省长白山国家级自然保护区阔叶红松林和针叶林中的黑熊(Ursus thibetanus)及棕熊(U.arctos)的相对种群数量和幼年个体比例进行了长期调查。调查面积分别为4万和3万hm2。调查期间累计遇到黑熊65头次(2.6头/年)、棕熊46头次(1.8头/年)。各年度黑熊和棕熊的数量变动很大,总体呈下降趋势。黑熊的数量较20世纪80年代下降了93.4%,幼体所占比率从17.2%下降为0%;棕熊种群数量下降了38.8%,幼体所占比率仅为7.1%。黑熊和棕熊的种群幼体所占比例极低,说明长白山自然保护区黑熊和棕熊自然繁殖力很低,2种熊种群均处于极度濒危状态。栖息地减少和盗猎可能是导致熊类数量急剧下降的主要因素。     

Seasonal distribution and food habits of spectacled Bears Tremarctos ornatus in the highlands of Ecuador

The seasonal distribution and food habits of the Spectacled Bear, Tremarctos ornatus, were studied in the subalpine paramo of the Antisana volcano, in Ecuador. The location and age of bear signs indicated that bears occupy paramo areas seasonally up to a maximum of 4,000 m from February to July. Feeding evidence and scat analysis confirmed that bromeliads (Puya sp.) are important bear's food in the highlands. Because the importance of paramo and montane forest habitats to the spectacled bear, large, contiguous areas of these ecotypes need to be preserved.     

Hemoglobin of tree sparrows (Passer montanus, Passeriformes): Sequence of the major (Hb A) and minor (Hb D) components

Blood of the adult Tree Sparrow (Passer montanus) contains two hemoglobin components, Hb A (ca. 85%), Hb D (ca. 15%). They differ in their alpha-chains (alpha A, alpha D), the beta-chains are identical. The complete primary structures of alpha A-, alpha D- and beta-chains are presented. Comparison with the Greylag Goose (Anser anser) hemoglobin (Hb A) showed that the alpha A-chains differ by 22 amino-acid exchanges, the beta-chains by 16. Comparison with the minor component of the Pheasant (Phasianus colchicus colchicus) hemoglobin (Hb D) showed that the alpha D-chains differ by 34 amino-acid exchanges. Proline is found incorporated in an internal position of an alpha-helix (pos. 124, H7). In comparison to that of the Starling (Sturnus vulgaris) the ratio of amino-acid exchanges for beta: alpha A: alpha D chains is 1 : 7 : 4; in comparison to other birds this ratio is found to be 1 : 2 (1.4-2.2):3 (2.2-4).     

Tubulopapillary mammary carcinoma in a brown bear (Ursus arctos)

A 28-yr-old, nulliparous female brown bear (Ursus arctos) at the Karacabey Ovakurusu Bear Sanctuary presented with an enlargement of the mammary gland. Three other nodules were also noted in the proximity of the mammary gland and over the vulva. Clinical, hematologic, ultrasonographic, and radiologic examinations were performed; the enlarged mammary gland was removed and the other masses were also excised. Histopathologic examination revealed tubulopapillary carcinoma of the mammary gland, and the other masses were diagnosed as epidermoid cysts. This is the first reported case of tubulopapillary mammary carcinoma accompanied by epidermoid cysts in a bear.     

The primary structure of pale-throated three-toed sloth (Bradypus tridactylus, Xenarthra) hemoglobin

The hemoglobin of the Pale-Throated Three-Toed Sloth (Bradypus tridactylus, Xenarthra) was separated into two components (ratio 4:1) with identical amino-acid analyses for the alpha- and beta-chains. The primary structures of both chains from the major component are given. They could be isolated by chromatography on carboxymethyl cellulose CM-52. The sequences have been determined by automatic Edman degradation of the native chains and their tryptic peptides. The comparison with human hemoglobin showed 27 substitutions in the alpha-chains and 33 in the beta-chains. In the alpha-chains one amino-acid exchange involves an alpha 1/beta 1-contact. In the beta-chains two heme- and four alpha 1/beta 1-contacts are substituted. The hemoglobin of the Sloth is compared to that of the Nine-Banded Armadillo (Dasypus novemcinctus), another representative of the order Xenerthra.     

Hemoglobin of the adult white stork (Ciconia ciconia, ciconiiformes). The primary structure of alpha A- and beta-chains from the only present hemoglobin component

The hemolysate obtained from erythrocytes of the adult White Stork (Ciconia ciconia) contains only one hemoglobin component, identified to be HbA. The complete primary structures of alpha A- and beta-chains are presented. The minor hemoglobin component HbD with alpha D-chains usually present in adult avian species was not detected by the White Stork. The sequence was determined by automatic Edman degradation of tryptic peptides and in the case of beta-chains additionally of the C-terminal peptide obtained by chemical cleavage at the Asp-Pro bond. Homologous comparison with the Greylag Goose (Anser anser) hemoglobin showed that the alpha A-chains differ by 23 amino-acid exchanges, the beta-chains by 17. Four of the substitutions in the alpha A-chains are in the alpha 1 beta 1-contact points, one in the alpha 1 beta 2-contacts and one in the amino acids near the heme. The amino-acid substitutions of the White Stork hemoglobin as compared to the other avian hemoglobins are discussed. We suggest that alpha D-chain is persistence of an embryonic gene.     

The primary structure of alpha A- and beta-chains from blue-and-yellow macaw (Ara ararauna, Psittaci) hemoglobin. No evidence for expression of alpha D-chains

The hemoglobin A of the Blue-and-Yellow Macaw (Ara ararauna) was isolated and characterized. The complete amino-acid sequence of alpha A- and beta-chains is presented. In contrast to some adult avian hemoglobins already investigated, Blue-and-Yellow Macaw hemoglobin is homogenous and contains only one component: HbA. The minor component, HbD, which is usually present in the hemolysate of avian erythrocytes, could not be detected. There is no evidence for the expression of the alpha D-globin gene. Comparison of alpha A- and beta-chains from Blue-and-Yellow Macaw hemoglobin with corresponding chains from Greylag Goose hemoglobin shows 19 amino-acid exchanges between alpha A-chains and 6 between beta-chains. The structure-function relationships of hemoglobin chains and the evolutionary aspects are discussed in view of these results.     

Carnivora: the primary structure of hemoglobin from the Masked Palm Civet (Paguma larvata, Viverridae)

The primary structure of the alpha- and beta-chains of hemoglobin from the Masked Palm Civet (Paguma larvata, Viverridae) is described. The chains were separated directly from hemoglobin by RP-HPLC. After tryptic digestion of the chains, the peptides were separated by RP-HPLC. Amino acid sequences were determined by Edman degradation in liquid and gas-phase sequencers. The alignment of the tryptic peptides was made by homology with human and other Carnivora hemoglobins. Paguma and human hemoglobin differ with respect to 23 amino-acid residues. Some of these amino-acid substitutions, which occur in both the alpha- and beta-chains, occur at contact sites between the subunits, and at the binding sites of heme and of organic phosphate, as well as involving residues responsible for the alkaline Bohr effect.     

The first sequenced normal hemoglobin lacking histidine in position 146 of the beta-chains. The primary structures of the major and minor hemoglobin components of the great crested newt (Triturus cristatus, Urodela, Amphibia)

The hemoglobin of the Great Crested Newt (Triturus cristatus), an animal maintaining the gas exchange to about 85% through the skin, consists of a major (HbM = 65%) and a minor (Hbm = 35%) component. The primary structures of the four chains are presented. They could be separated by reversed-phase HPLC and were cleaved with trypsin and additionally by acid hydrolysis. Both the native chains and their peptides were sequenced by liquid and gas phase sequenators. At the N-terminus the alpha M-chains are by one amino-acid residue longer and the beta M-chains by one residue shorter, resulting in a chain length of 142 and 145, respectively. The alpha m-chains are of normal length whereas in the beta m-chains the C-terminal histidine in position 146 is missing. Both alpha-chains differ by 50 residues (35.2%) and the beta-chains by 63 (43.2%). The alpha-chains were compared with those of other salamandroid hemoglobins. The difference to human hemoglobin is marked by 61 (43.3%) amino-acid substitutions in both alpha-chains and by 78 (53.4%) in both beta-chains. Numerous heme contacts and positions involved in the subunit interface are affected by replacements. The most interesting of them were studied by molecular modeling. The importance of the missing beta m-146(HC3)His and of the substitution of several amino-acid residues involved in the binding of organic phosphates is discussed with respect to the reduced Bohr effect of Triturus cristatus hemoglobin.     

The primary structure of the hemoglobin of the dogfish shark (Squalus acanthias). Antagonistic effects of ATP and urea on oxygen affinity of an elasmobranch hemoglobin

The amino-acid sequence of the hemoglobin of the Dogfish Shark (Squalus acanthias) is presented. The alpha-chains consist of 141 residues and show a Thr/Ser ambiguity at position 3. The beta-chains consist of 142 residues and evidently have no D-helix; they show an Asn/Tyr ambiguity at position 104. Both chains have free N-terminal amino acids. The phylogenetic distance from the human alpha- and beta-chains is indicated by 49.3% and 56.2% amino-acid exchanges. The primary structure is discussed in relation to the oxygen-binding properties of elasmobranch hemoglobin, particularly as regards the antagonistic effects of urea and ATP, and the effects of proton concentration (the alkaline and acid Bohr effects, and the Root effect).     

Efficacy of Bear Deterrent Spray in Alaska

Abstract: We present a comprehensive look at a sample of bear spray incidents that occurred in Alaska, USA, from 1985 to 2006. We analyzed 83 bear spray incidents involving brown bears (Ursus arctos; 61 cases, 74%), black bears (Ursus americanus; 20 cases, 24%), and polar bears (Ursus maritimus; 2 cases, 2%). Of the 72 cases where persons sprayed bears to defend themselves, 50 (69%) involved brown bears, 20 (28%) black bears, and 2 (3%) polar bears. Red pepper spray stopped bears' undesirable behavior 92% of the time when used on brown bears, 90% for black bears, and 100% for polar bears. Of all persons carrying sprays, 98% were uninjured by bears in close-range encounters. All bear—inflicted injuries (n = 3) associated with defensive spraying involved brown bears and were relatively minor (i.e., no hospitalization required). In 7% (5 of 71) of bear spray incidents, wind was reported to have interfered with spray accuracy, although it reached the bear in all cases. In 14% (10 of 71) of bear spray incidents, users reported the spray having had negative side effects upon themselves, ranging from minor irritation (11%, 8 of 71) to near incapacitation (3%, 2 of 71). Bear spray represents an effective alternative to lethal force and should be considered as an option for personal safety for those recreating and working in bear country. (JOURNAL OF WILDLIFE MANAGEMENT 72(3):640–645; 2008)     

The amino-acid sequence of Northern Mallard (Anas platyrhynchos platyrhynchos) hemoglobin

The complete amino-acid sequence of the major hemoglobin component (HbA) of the adult Northern Mallard (Anas platyrhynchos platyrhynchos) is presented. A minor component HbD was also detected but in low concentrations. The sequences of alpha A- and beta-chains were established by automatic Edman degradation on tryptic peptides and peptides obtained by specific chemical cleavages. The alignment of the peptides was performed by comparison with the alpha A- and beta-chains of Greylag Goose hemoglobin (Anser anser). Thereby an exchange of five positions in the alpha A-chains and three in the beta-chains was observed. No exchanges were found in the surroundings of the heme, in alpha 1 beta 2-contact points, or allosteric regulatory sites. In the alpha 1 beta 1-subunit interface one amino-acid residue in alpha A-chains and one in beta-chains are exchanged. By comparison with the amino-acid sequence derived from globin genes of Domestic Duck (Anas platyrhynchos), the alpha A-chains differ by two exchanges in the N-terminal region and the beta-chains by five exchanges the in C-terminal region. The comparison of the amino-acid sequence derived from alpha A-globin gene of the Moscovy Duck (Cairina moschata) and alpha A-chains of the Northern Mallard, shows only one replacement.     

The primary structure of the hemoglobin of the Cormorant (Phalacrocorax carbo, Pelecaniformes)

The erythrocytes of the adult Cormorant contain two hemoglobin components in a ratio of 83% Hb A to 17% Hb D. The primary structures of the alpha A-, alpha D- and beta-chains are presented. The globin chains were separated by high-performance liquid chromatography and cleaved enzymatically and/or chemically. The native chains and their fragments were sequenced using liquid- or gas-phase sequencers, and the peptides aligned using the homology to human and to avian hemoglobin sequences. Compared to human hemoglobin, there are 46 amino-acid replacements in the alpha A-chains (67.4% homology), 65 replacements in the alpha D-chains (53.9% homology) and 45 replacements in the beta-chains (69.2% homology). In the functionally important regions, the percentage of amino-acid substitutions, as compared to human hemoglobin, is 13.2% in the alpha A-, 19.0% in the alpha D - and 16.0% in the beta-chains. The importance of the replacement beta 135 arginine (other birds)----glycine (Cormorant) in the phosphate-binding pocket and its effect on phosphate binding will be discussed.     

Carnivora: the primary structure of the common otter (Lutra lutra, Mustelidae) hemoglobin

The hemoglobin of the Common Otter (Lutra lutra, Carnivora) contains only one component. The complete primary structures of the alpha- and beta-chains are presented. They were separated by high-performance liquid chromatography and the sequences determined by automatic liquid and gas-phase Edman degradation of the chains and their tryptic peptides. The alpha-chains show 18 and the beta-chains 13 substitutions compared to human alpha- and beta-chains, respectively. In the alpha-chains one heme- and two alpha 1/beta 1-contacts are exchanged. In the beta-chains the replacements involve one heme-, one alpha 1/beta 1-, and one alpha 1/beta 2-contact. The alpha- and beta-chains of the Common Otter are compared to those of other Carnivora hemoglobins. The unexpected low number of substitutions between Common Otter hemoglobin and that of Lesser Panda as well as of Harbor Seal is discussed.