Crystallization, preliminary X-ray diffraction analysis and Patterson search of oxyhaemoglobin I from the wolf (Chrysocyon brachiurus)

Oxyhaemoglobin I isolated from the Brazilian wolf Chrysocyon brachiurus has been crystallized and X-ray diffraction data has been collected to 2.06 A resolution using a synchrotron-radiation source. Crystals were determined to belong to the space group P2(1)2(1)2(1) and preliminary structural analysis revealed the presence of one tetramer in the asymmetric unit. The structure was determined using standard molecular-replacement techniques and is currently being refined using maximum-likelihood protocols. This is the first haemoglobin isolated from a member of the Canidae family to be crystallized and it will provide further insights in the comparative biochemistry of vertebrate haemoglobins.

Ablação por cateter versus tratamento farmacológico para pacientes com síndrome de Wolf-Parkinson-White: revisão sistemática

Pós-graduação em Bases Gerais da Cirurgia - FMB

Wolf presence in the ranch of origin: Impacts on temperament and physiological responses of beef cattle following a simulated wolf encounter

This experiment evaluated temperament, vaginal temperature, and plasma cortisol in beef cows from wolf-naive and wolf-experienced origins that were subjected to a simulated wolf encounter. Multiparous, pregnant, nonlactating Angus-crossbreed cows from the Eastern Oregon Agricultural Research Center located near Burns, OR (CON; n = 50), and from a commercial operation near Council, ID (WLF; n = 50), were used. To date, grey wolves are not present around Burns, OR, and thus CON were naive to wolves. Conversely, wolves are present around Council, ID, and WLF cows were selected from a herd that had experienced multiple confirmed wolf-predation episodes from 2008 to 2012. Following a 50-d commingling and adaptation period, CON and WLF cows were ranked by temperament, BW, and BCS and allocated to 5 groups (d 0; 10 CON and 10 WLF cows/group). Groups were individually subjected to the experimental procedures on d 2 (n = 3) and d 3 (n = 2). Before the simulated wolf encounter, cow temperament was assessed and blood samples and vaginal temperatures (using intravaginal data loggers) were collected (presimulation assessments). Cows were then sorted by origin, moved to 2 adjacent drylot pens (10 WLF and 10 CON cows/pen), and subjected to a simulated wolf encounter event for 20 min, which consisted of 1) cotton plugs saturated with wolf urine attached to the drylot fence, 2) continuous reproduction of wolf howls, and 3) 3 leashed dogs that were walked along the fence perimeter. Thereafter, WLF and CON cows were commingled and returned to the handling facility for postsimulation assessments, which were conducted immediately after exposure to wolf-urine-saturated cotton plugs, wolf howl reproduction, and 20-s exposure to the 3 dogs while being restrained in a squeeze chute. Chute score, temperament score, and plasma cortisol concentration increased (P <= 0.01) from pre- to postsimulation assessment in WLF but did not change in CON cows (P >= 0.19). Exit velocity decreased (P = 0.01) from pre-to postsimulation assessment in CON but did not change (P = 0.79) in WLF cows. In addition, WLF cows had a greater (P = 0.03) increase in temperature from pre-to postsimulation assessments compared with CON cows. In conclusion, the simulated wolf encounter increased excitability and fear-related physiological stress responses in cows that originated from a wolf-experienced herd but not in cows that originated from a wolf-naive herd.

Genetic differentiation among distinct karyomorphs of the wolf fish Hoplias malabaricus species complex (Characiformes, Erythrinidae) and report of unusual hybridization with natural triploidy

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

A transitoriedade entre o mundo primitivo e o moderno nas obras The Sea-Wolf e White Fang de Jack London

The American author Jack London wrote the books The Sea-Wolf and White Fang in 1904 and 1906, respectively. The former portrays the life of a literary critic, Humphrey Van Weyden, who after the wreckage of the ship in which he was is rescued by a seal-hunting schooner, where he is obliged to work and to live with the brutal captain Wolf Larsen, and in doing so he develops a more primitive profile. The latter work portrays the life of the eponymous character, a wolf that leaves the wild world in which it was born to follow its last master in the city, thus developing features of the modern world. This paper aims to conduct an analysis of the aspects that allow the transition between the primitive and the modern world in those works

Management of Wolf-Livestock Conflicts in Minnesota

In 1975, the gray wolf (Canis lupus) population in Minnesota was protected by the federal Endangered Species Act (USA). At that time, there were 500-750 wolves. By 2004, the population had grown to an estimated 3,020 wolves. Over time, conflicts between wolves and livestock increased. Wolf depredation control programs have been conducted by the U.S. Fish and Wildlife Service (1975-1986) and by the U.S. Department of Agriculture’s Wildlife Services program (1986 to present). In 1978, Minnesota’s wolves were reclassified from endangered to threatened which allowed authorized federal agents to lethally remove wolves that had depredated on livestock or pets. A State funded wolf compensation program was also established in 1978. Wildlife Services’ wolf damage management approach utilizes both nonlethal and lethal methods of control. Currently, wolf depredations are verified at 60-85 farms annually and 125-175 wolves are taken each year. Wolf compensation payments to livestock producers have averaged $67,111 per year during the past five years. Most livestock losses occur during spring and summer. Selective removal of depredating wolves, coupled with improvements in animal husbandry practices, has potential for reducing wolf-livestock conflicts. Minnesota’s wolf population is currently considered to be fully recovered and federal delisting is expected to occur in the near future.

The Effects of Ornamentation on Wolf Spider Foraging

ABSTRACT: As a secondary sexual trait that can increase reproductive success, ornamentation is believed to be costly for organisms to acquire and possess. This study investigates possible costs of ornamentation on wolf spider foraging by comparing foraging abilities of two male forms that differ in ornamentation upon maturation. The two male forms, found syntopically in a mixed population in Mississippi, USA, resemble two sibling species: Schizocosa ocreata, in which males develop large black brushes on their forelegs upon maturation, and Schizocosa rovneri, in which males lack ornamentation following maturation. Individuals of both forms participated in foraging trials as penultimates (juveniles) and as matures. Analyses were conducted to compare behaviors and determine changes in foraging abilities between male forms (non-ornamented vs. brush-legged) and between age groups (penultimate vs. mature). Most foraging behaviors of the two male forms during immaturity were similar with the exception that brush-legged males attacked more frequently than non-ornamented males. Brush-legged males attacked less, spent more time moving, and improved capture abilities as matures, while non-ornamented males retained similar trends for these behaviors with age. Additionally, while capture abilities improved with age among brush-legged males, killing abilities remained constant. This disparity was due to increased escapes made by captured prey items, possibly due to hindrances caused by brush presence preventing secure holds onto crickets. In summary, differences in foraging exist between brush-legged and non-ornamented males prior to sexual maturation, and the development of/presence of brushes appears to influence adult male foraging efficiency.

WOLF CONTROL IN BRITISH COLUMBIA, CANADA

Up to 1949, the Fish and Game Branch employed personnel, some of whom were temporary, to attempt control of the extremely high wolf pop¬ulations of the central and northern portions of British Columbia. Coyotes were also very numerous in the central and southern regions and had to be considered because of their depredations. The field men were keen and conscientious but their efforts were not co-ordinated. Control areas were severely restricted in size as techniques were not adaptable enough and because of a lack of manpower. Eventually, sheepmen went out of business entirely over wide areas, cattlemen were subjected to huge annual losses, and sportsmen were very concerned. However, stock losses constituted the major complaint and resulted in ranchers demanding action* Two major changes came out of this. First, the bounty on wolves was raised and second, the present Predator Control Division was formed. The administration was convinced that a force of experienced, fully-trained field staff under a single supervision would be far more effective than bounty payments. Unfortunately, bounties were in vogue during that time and forced the necessity of proving the worth of organized controls before any consideration could be given to the elimination of the bounty system.

Genomic characterization of the italian wolf (Canis lupus): the genes involved in black coat colour determination and application of microarray technique for snps detection.

This study provides a comprehensive genetic overview on the endangered Italian wolf population. In particular, it focuses on two research lines. On one hand, we focalised on melanism in wolf in order to isolate a mutation related with black coat colour in canids. With several reported black individuals (an exception at European level), the Italian wolf population constituted a challenging research field posing many unanswered questions. As found in North American wolf, we reported that melanism in the Italian population is caused by a different melanocortin pathway component, the K locus, in which a beta-defensin protein acts as an alternative ligand for the Mc1r. This research project was conducted in collaboration with Prof. Gregory Barsh, Department of Genetics and Paediatrics, Stanford University. On the other hand, we performed analysis on a high number of SNPs thanks to a customized Canine microarray useful to integrate or substitute the STR markers for genotyping individuals and detecting wolf-dog hybrids. Thanks to DNA microchip technology, we obtained an impressive amount of genetic data which provides a solid base for future functional genomic studies. This study was undertaken in collaboration with Prof. Robert K. Wayne, Department of Ecology and Evolutionary Biology, University of California, Los Angeles (UCLA).

Molekulare Untersuchung des Wolf-Hirschhorn-Syndroms

Das Wolf-Hirschhorn-Syndrom (WHS) ist ein komplexes und variables Fehlbildungs- Retardierungssyndrom, das durch Deletion in der distalen Chromosomenregion 4p16.3 hervorgerufen wird und dessen Ätiologie und Pathogenese bisher weitgehend unverstanden sind. Die Zielsetzung in der vorliegenden Arbeit bestand in der Identifizierung und vorläufigen Charakterisierung neuer Gene, die an der Entstehung des Syndroms beteiligt sein könnten. Die Wolf-Hirschhorn-Syndrom-kritische Region (WHSCR) konnte zu Beginn der vorliegenden Arbeit auf einen ca. 2 Mb großen Bereich zwischen den Markern D4S43 und D4S142 eingegrenzt werden. Für die Identifizierung neuer Gene wurden zunächst drei größere genomische Cosmid-/PAC-Contigs (I-III) im Bereich der Marker D4S114 bis D4S142 erstellt und mittels Exonamplifikation auf transkribierte Bereiche (Exons) untersucht. Es konnten insgesamt 67 putative 'Exons' isoliert werden, von denen einige bereits bekannten Genen (ZNF141, PDEB, MYL5, GAK, DAGK4 und FGFR3) entsprechen. Zwei dieser Gene konnten im Rahmen dieser Arbeit erstmals (DAGK4) bzw. genauer (GAK) in die distale Region 4p16.3 kartiert werden. Die restlichen Exons können aufgrund von Homologievergleichen und/oder EST-cDNA-Homologien vermutlich neuen Genen oder auch Pseudogenen (z. B. YWEE1hu) zugeordnet werden. Durch die im Verlaufe der vorliegenden Arbeit publizierte weitere Eingrenzung der WHSCR auf einen 165 Kb-großen Bereich proximal des FGFR3-Gens konzentrierten sich weitere Untersuchungen auf die detaillierte Analyse der WHSCR zwischen dem Marker D4S43 und FGFR3. Mit Hilfe von Exonamplifikation bzw. computergestützter Auswertung vorliegender Sequenzdaten aus diesem Bereich ('GRAIL', 'GENSCAN' und Homologievergleiche in den EST-Datenbanken des NCBI) konnten mehrere neue Gene identifiziert werden. In distaler-proximaler Reihenfolge handelt es sich dabei um die Gene LETM1, 51, 43, 45, 57 und POL4P. LETM1 kodiert für ein putatives Transmembran-Protein mit einem Leucin-Zipper- und zwei EF-Hand-Motiven und könnte aufgrund seiner möglichen Beteiligung an der Ca2+-Homeostase und/oder der Signal-transduktion zu Merkmalen des WHS (Krampfanfällen, mentale Retardierung und muskuläre Hypotonie) beitragen. Das Gen 51 entspricht einem in etwa zeitgleich durch Stec et al. (1998) und Chesi et al. (1998) als WHSC1 bzw. MMSET bezeichnetem Gen und wurde daher nicht weiter charakterisiert. Es wird genauso wie das Gen 43, das zeitgleich von Wright et al. (1999b) als WHSC2 beschrieben werden konnte und eine mögliche Rolle bei der Transkriptionselongation spielt, ubiquitär exprimiert. Das in der vorliegenden Arbeit identifizierte Gen 45 zeigt demgegenüber ein ausgesprochen spezifisches Expressionsmuster (in Nervenzellen des Gehirns sowie in Spermatiden). Dies stellt zusammen mit der strukturellen Ähnlichkeit des putativen Genprodukts zu Signalmolekülen einen interessanten Zusammenhang zu Merkmalen des WHS (beispielsweise Kryptorchismus, Uterusfehlbildungen oder auch neurologische Defekte) her. Demgegenüber handelt es sich bei dem Gen 57 möglicherweise um ein trunkiertes Pseudogen des eRFS-Gens auf Chromosom 6q24 (Wallrapp et al., 1998). Das POL4P-Gen schließlich stellt allein aufgrund seiner genomischen Lokalisation sowie seiner möglichen Funktion (als DNA-Polymerase-ähnliches Gen) kein gutes Kandidatengen für spezifische Merkmale des Syndroms dar und wurde daher nicht im Detail charakterisiert. Um die Beteiligung der Gene an der Ätiologie und Pathogenese des Syndroms zu verstehen, ist die Entwicklung eines Mausmodells (über das Einfügen gezielter Deletionen in das Mausgenom) geplant. Um dies zu ermöglichen, wurde in der vorliegenden Arbeit die Charakterisierung der orthologen Region bei der Maus vorgenommen. Zunächst wurden die orthologen Gene der Maus (Letm1, Whsc1, Gen 43 (Whsc2h), Gen 45 und Pol4p) identifiziert. Durch die Erstellung sowie die genaue Kartierung eines murinen genomischen P1/PAC-Klon-Contigs konnte gezeigt werden, daß die murinen Gene Fgfr3, Letm1, Whsc1, Gen 43 (Whsc2h), Gen 45 und Pol4p sowie einige weitere der überprüften EST-cDNA-Klone der Maus in einem durchgehenden Syntänieblock zwischen Mensch (POL4P bis FGFR3) und Maus (Mmu 5.20) enthalten sind, der in seiner genomischen Ausdehnung in etwa den Verhältnissen beim Menschen (zwischen POL4P und FGFR3) entspricht.

Ecological genetics and conservation genomics of wolf (Canis lupus)

In the present work, we apply both traditional and Next Generation Sequencing (NGS) tools to investigate some of the most important adaptive traits of wolves (Canis lupus). In the first part, we analyze the variability of three Major Histocompatibility Complex (MHC) class II genes in the Italian wolf population, also studying their possible role in mating choice and their influence on fitness traits. In the second section, as part of a larger canid genome project, we will exploit NGS data to investigate the transcript-level differences between the wolf and the dog genome that can be correlated to domestication.

Landscape ecology and genetics of the wolf in Italy

This PhD Thesis includes five main parts on diverse topics. The first two parts deal with the trophic ecology of wolves in Italy consequently to a recent increase of wild ungulates abundance. Data on wolf diet across time highlighted how wild ungulates are important food resource for wolves in Italy. Increasing wolf population, increasing numbers of wild ungulates and decreasing livestock consume are mitigating wolf-man conflicts in Italy in the near future. In the third part, non-invasive genetic sampling techniques were used to obtain genotypes and genders of about 400 wolves. Thus, wolf packs were genetically reconstructed using diverse population genetic and parentage software. Combining the results on pack structure and genetic relatedness with sampling locations, home ranges of wolf packs and dispersal patterns were identified. These results, particularly important for the conservation management of wolves in Italy, illustrated detailed information that can be retrieved from genetic identification of individuals. In the fourth part, wolf locations were combined with environmental information obtained as GIS-layers. Modern species distribution models (niche models) were applied to infer potential wolf distribution and predation risk. From the resulting distribution maps, information pastures with the highest risk of depredation were derived. This is particularly relevant as it allows identifying those areas under danger of carnivore attack on livestock. Finally, in the fifth part, habitat suitability models were combined with landscape genetic analysis. On one side landscape genetic analyses on the Italian wolves provided new information on the dynamics and connectivity of the population and, on the other side, a profound analysis of the effects that habitat suitability methods had on the parameterization of landscape genetic analyses was carried out to contributed significantly to landscape genetic theory.