Patterns of fecal gonadal hormone metabolites in the maned wolf (Chrysocyon brachyurus)

Ex situ populations of maned wolves are not viable due to low reproductive efficiency. The objective of this study was to increase knowledge regarding the reproductive physiology of maned wolves to improve captive management. Fecal samples were collected 3-5 d/wk from 12 females of various reproductive age classes (young, prime breeding and aged) and reproductive histories (conceived and raised pups, conceived but lost pups, pseudo-pregnant and unpaired). Ovarian steroids were extracted from feces and assessed by enzyme immunoassay. Concentrations of estrogen metabolites gradually increased, beginning 2-5 d before breeding, and declined to baseline on the day of lordosis and copulation. Fecal progestin metabolite concentrations increased steadily during the periovulatory period, when sexual receptivity was observed, and remained elevated during pregnancy and pseudo-pregnancy. During the luteal phase, young and prime breeding-age females excreted larger amounts of progestins than those of older age classes. Furthermore, progestin concentrations were higher during the luteal phase of pregnant versus pseudo-pregnant bitches. Profiles of fecal progestin metabolites for three singleton females were unchanged throughout the breeding season, suggesting ovulation is induced in this species. However, this finding could be confounded by age, as these females were either young or aged.     

Diet of the maned wolf, Chrysocyon brachyurus, in central Brazil

Analysis of 105 scats of the maned wolf ( Chrysocyon brachyurus ) collected in central Brazil yielded 304 occurrences of food items of which fruits of Solanum lycocarpum , rodents, and birds accounted for 61.5%. By analysis of minimum number of individual animals, rodents and birds were 75% of a total of 156 prey. On the other hand, fruits of S. lycocarpum and armadillos ( Dasypus spp.) were the bulk of the total estimated biomass consumption, yielding 63.7% in a total of 73.5 kg. Rodents were mostly captured during the dry season, while miscellaneous fruits were consumed mostly in the wet season. The consumption of S. lycocarpum fruits and armadillos was aseasonal. Small rodents were taken in about the ranks of abundance in the study area, but S. lycocarpum fruits were actively searched by maned wolves, for its occurrence is limited to secondary savanna. Prey and fruits typical of savanna ('cerrado') and grassland ('campo') were the bulk of species, occurrences, prey numbers, and biomass consumed. These findings reinforce the importance of conservation of cerrado and campo in central Brazil. Despite maned wolf being a vulnerable species, its feeding habits are opportunistic, and the main reasons for its decline are likely to be habitat destruction and human disturbance.     

Diet of the maned wolf (Chrysocyon brachyurus) and its role in seed dispersal on a cattle ranch in Brazil

The maned wolf Chrysocyon brachyurus is the largest canid inhabiting South America. Its geographic distribution includes the open fields of Brazil's central area, which is currently undergoing agricultural expansion. The diet of the maned wolf and its seasonal variation was determined on a dairy cattle ranch (São Luís farm, 566 ha) in the State of Minas Gerais, Brazil. From January to December faeces of the maned wolf were collected monthly ( n = 150 scats; 397 food item occurrences). Twenty-nine taxa were identified from scats, 18 of animal origin (46% or 183 occurrences) and 11 of plants (54% or 214 occurrences). The fruits of Solanum lycocarpum were the dominant food item in our study (29%). Mammals contributed 13%, arthropods 12%, birds 11% and reptiles 2% of the food items. Arthropods and fruits were prevalent in the rainy season and mammals in the dry season. As expected for a heavily farmed region, frugivory results were at the lower end of the diversity scale (9–33 species) and included four old garden species. No previous study of the diet of maned wolf has registered as many species of Solanaceae as this one. Although dietary richness was lower, the main food items (wolf fruit, armadillos, rodents, birds) were the same as study sites in 'cerrado' and upland meadows. In this region, the open habitats occupied by the maned wolf were previously covered by Atlantic forest, suggesting that landscape modification such as cattle ranching has opened new frontiers for distribution expansion of the maned wolf. The impact of loss of dietary richness and the increase in Solanaceae on the survival of the maned wolf need to be evaluated.     

Use of behavioral measures to assess reproductive status in maned wolves (Chrysocyon brachyurus)

Captive breeding efforts of the threatened maned wolf (Chrysocyon brachyurus) have been plagued by a lack of reliable reproduction and a high rate of neonatal mortality. A particular problem for animal managers has been the ability to detect pregnancy in a noninvasive manner. Pseudopregnancies are common, and many staff hours are expended preparing for a birth that may not occur. The objectives of our study were to document changes in behavior during the breeding season in captive maned wolves in order to determine if behaviors other than sexual (i.e., copulation) could be used to distinguish nonbreeding from breeding pairs and, further, whether successful breeding pairs (young born) could be distinguished behaviorally from breeding pairs that did not produce young (pseudopregnant). Between 1988 and 1994, behavioral data were collected during the annual reproductive season from 52 maned wolf pairings (27 males, 26 females) housed at 17 North American institutions. Breeding animals showed significant increases in rates of affiliative behaviors (approach, friendly) and the amount of time spent in close association (social) during the estrous period, compared to pre- and post-estrous periods. In contrast, the behavior of nonbreeding pairs varied little throughout the breeding season. Discriminant function analysis (DFA) showed that several behavioral measurements during certain reproductive periods were useful in discriminating between nonbreeding and breeding animals (rates of friendly and social behavior for males, marking and social behavior for females). Unsuccessful (pseudopregnant) and successful breeding pairs could be further distinguished on the basis of agonistic and approach behavior frequencies (successful females showed higher rates of agonistic and lower rates of approach behavior during certain periods; successful males had higher rates of both behaviors). Discriminant equations for key behavioral measurements and examples of their practical application are presented. DFA provides animal managers with an effective, noninvasive technique for assessing the reproductive status of maned wolf pairs. © 1996 Wiley-Liss, Inc.     

Secret lives of maned wolves (Chrysocyon brachyurus Illiger 1815): as revealed by GPS tracking collars

The maned wolf Chrysocyon brachyurus is a nocturnal and shy species, which has proven difficult to study in the field; consequently, data about its behavioural biology are almost absent from the scientific literature. However, recent advances in global positioning system (GPS) technology mean that it is now possible to study the socio-spatial dynamics of this species in the field. In the present study three related maned wolves (an adult pair and their juvenile female offspring) were monitored by GPS tracking collars for a 6-month period, which included the birth of a litter of pups to the adult pair. The three GPS collars were programmed to record the position of the wearer every 2 h (simultaneously for all three individuals). Analyses of the data from the three individuals showed that the female had the largest home range, then the male and finally the juvenile. Furthermore, there was considerable overlap in home-range use by all three individuals. The home ranges of the adult pair also varied in relation to the birth of pups (decreasing dramatically and then showing a slow increase). During the night, which is the normal period of activity for this species, the three individuals avoided contact with each other. However, during the day, the adult pair usually slept together and their juvenile slept within a few hundred metres of their location. The distance travelled at night was greatest for the female, then the male and finally the juvenile. These data suggest that (1) the social bond between the male and female is strong, (2) maned wolves are highly tolerant of their juvenile offspring, even in the case of new pups being born, and (3) that maned wolves do not hunt together and, in fact, avoid each other when hunting.     

Evaluation of homologous, heterologous, and affinity conjugates for the serodiagnosis of Toxoplasma gondii and Neospora caninum in maned wolves (Chrysocyon brachyurus)

Use of serological tests in the diagnosis of infectious diseases in wild animals has several limitations, primarily the difficulty of obtaining species-specific reagents. Wild canids, such as maned wolves (Chrysocyon brachyurus), are highly predisposed to infection by Toxoplasma gondii and, to a lesser extent, to Neospora caninum. The aim of the present study was to evaluate homologous, heterologous, and affinity conjugates in enzyme-linked immunosorbent assays (ELISAs) and indirect fluorescent antibody tests (IFATs) for detecting immunoglobulin (Ig) G antibodies against T. gondii and N. caninum in maned wolves. Serum samples were obtained from 59 captive animals in Brazil and tested by ELISA for T. gondii serology and IFAT for N. caninum serology using 3 different enzymatic and fluorescent conjugates: homologous (guinea pig anti-maned wolf IgG-peroxidase and -fluorescein isothiocyanate [FITC]), heterologous (rabbit anti-dog IgG-peroxidase and -FITC), and affinity (protein A-peroxidase and -FITC). Seropositivity to T. gondii was comparable among the homologous (69.5%), heterologous (74.6%), and affinity (71.2%) enzymatic conjugates. A significant positive correlation was found between the antibody levels determined by the 3 enzymatic conjugates. The highest mean antibody levels (ELISA index = 4.5) were observed with the protein A-peroxidase conjugate. The same seropositivity to N. caninum (8.5%) was found with the homologous and heterologous fluorescent conjugates, but protein A-FITC was not able to detect or confirm any positive samples with homologous or heterologous conjugates. Our results demonstrate that homologous, heterologous, and affinity conjugates might be used in ELISA for serological assays of T. gondii in wild canids, whereas for N. caninum infection, only the homologous or heterologous fluorescent conjugates have been shown to be useful.     

Crystallographic structure analysis of lamprey hemoglobin from anomalous dispersion of synchrotron radiation

The molecular structure of lamprey hemoglobin was previously determined and refined by conventional crystallographic analysis. In this study, the structural analysis has been repeated in the course of developing the method of multiwavelength anomalous diffraction (MAD) for phase determination. New experimental and analytical procedures that were devised to perform this determination should have general applicability. These include an experimental design to optimize signal strength and reduce systematic errors, experimental evaluation of anomalous scattering factors, and a least-squares procedure for analyzing the MAD data. MAD phases for the structure at 3 A resolution are as accurate overall as the multiple isomorphous replacement (MIR) phases determined previously.     

Modelling potential impacts of climate change on the bioclimatic envelope and conservation of the Maned Wolf (Chrysocyon brachyurus)

Forecasting the influence of climatic changes on the distribution of the Maned Wolf (Chrysocyon brachyurus) is important for the conservation of the species. We explored the environmental characteristics than best explain the current distribution of the species, modelled the past and present distribution, projected the niche model into the future, and identified suitable areas for conservation. Niche modelling was performed using Maxent and 21 environmental variables. For past conditions, we considered the Last Glacial Maximum (LGM) and the mid-Holocene (MH) climates. For future conditions, we used the A2a greenhouse gas emission scenario for 2050. Four General Circulation Models (FGOALS 1.0, HADCM3, IPSL-CM4 and MIROC 3.2) were used. The resulting niche model (AUC = 0.89 ± 0.02) predicts maximum probability of presence at precipitation of 106 mm during the coldest quarter, of 396 mm during the warmest quarter, and in totally flat areas. The suitable area for the Maned Wolf currently covers 4,320,364 km². For the LGM, there were inter-model differences in predicted areas (from 819,324 km² to 6,395,886 km²) and in geographic location. The MH models showed drastic changes with respect to the present and considerable inter-model variation. Predictions for 2050 show significant (at least 33%) reductions in distribution. Only a minor fraction (39%) of the current distribution can be considered stable for the period LGM-2050. The FGOALS model was the best option for projecting species occurrence into the future because it included the three localities known for the Maned Wolf from the late Pleistocene and predicts stable areas that coincide with spatial patterns of genetic diversity. The FGOALS projection for 2050 predicts a 33% reduction in suitable habitats, indicating some stable areas (central South America) that will probably be key sites for the conservation of the species.     

Capillariidae eggs found in the urine of a free ranging maned wolf from Argentina

The first finding of a Capillariid in the urinary tract of a free ranging maned wolf (Chrysocyon brachyurus) is described. The individual was an adult male attacked by dogs in the locality of Cayastacito (Santa Fe, Argentina, 31 degrees 05' S, 60 degrees 34' W). Eggs found in urine measured 64.6-66.9 micrometer (mean 65.4 micrometer) x 26.9-31 micrometer (mean 29 micrometer). Further studies are needed to determine whether this finding corresponds to a new Capillariid species, related to C. brachyurus, or it is an already described species that has been introduced by domestic dogs.     

Medical physics aspects of the synchrotron radiation therapies: Microbeam radiation therapy (MRT) and synchrotron stereotactic radiotherapy (SSRT)

Stereotactic Synchrotron Radiotherapy (SSRT) and Microbeam Radiation Therapy (MRT) are both novel approaches to treat brain tumor and potentially other tumors using synchrotron radiation. Although the techniques differ by their principles, SSRT and MRT share certain common aspects with the possibility of combining their advantages in the future. For MRT, the technique uses highly collimated, quasi-parallel arrays of X-ray microbeams between 50 and 600 keV. Important features of highly brilliant Synchrotron sources are a very small beam divergence and an extremely high dose rate. The minimal beam divergence allows the insertion of so called Multi Slit Collimators (MSC) to produce spatially fractionated beams of typically ∼25–75 micron-wide microplanar beams separated by wider (100–400 microns center-to-center(ctc)) spaces with a very sharp penumbra. Peak entrance doses of several hundreds of Gy are extremely well tolerated by normal tissues and at the same time provide a higher therapeutic index for various tumor models in rodents. The hypothesis of a selective radio-vulnerability of the tumor vasculature versus normal blood vessels by MRT was recently more solidified.SSRT (Synchrotron Stereotactic Radiotherapy) is based on a local drug uptake of high-Z elements in tumors followed by stereotactic irradiation with 80 keV photons to enhance the dose deposition only within the tumor. With SSRT already in its clinical trial stage at the ESRF, most medical physics problems are already solved and the implemented solutions are briefly described, while the medical physics aspects in MRT will be discussed in more detail in this paper.     

The complete primary structure of late lactation protein from quokka (Setonix brachyurus)

The complete primary structure of the late lactation protein from the milk of quokka (Setonix brachyurus) is presented. The amino acid sequence was established by N-terminal sequence analysis of high-performance liquid chromatography purified intact protein and peptides isolated from chemical and enzymatic digests of the protein. The protein contains 158 residues including four cysteines. The sequence comparison with the tamar wallaby (Macropus eugenii) late lactation protein shows only five differences. The protein is identified as a new member of a novel late lactation protein family present in the milk of marsupials.     

Camera-trap studies of maned wolf density in the Cerrado and the Pantanal of Brazil

The maned wolf (Chrysocyon brachyurus) is threatened by large-scale habitat loss, in particular due to conversion to agricultural land. This is the first published study on maned wolf density and the first test of individual identification from camera-trap photographs. We present results from two Brazilian regions: the Cerrado and the Pantanal. Using capture–recapture analysis of camera-trap data, we estimated densities per 100 square kilometers of 3.64 ± 0.77 individuals at the Cerrado site and 1.56 ± 0.77 individuals at the Pantanal site. Parallel radio-telemetry studies at the Pantanal site showed that maned wolves occupied home ranges of 39–58 km² (mean = 50.3 ± 7.67 km²). Our study in the Cerrado took place in a private farm with a mixture of agricultural land and native habitats, representative of the majority of the present-day Cerrado. Whereas many other mammalian species have suffered in the region, our results show that the maned wolf may cope better with this highly fragmented landscape than one might have feared. Finally, the paper briefly compares maned wolf density with density of puma (Puma concolor) in the Pantanal site.     

The tertiary structure of azurin from Pseudomonas denitrificans as determined by Cu resonant diffraction using synchrotron radiation

The structure of Pseudomonas denitrificans azurin has been solved at 3.0 A resolution by film data collection methods using synchrotron radiation. Bijvoet pairs of reflections were collected using 8.98 keV radiation where both delta f' and delta f", the real and imaginary corrections to azurin's Cu prosthetic group scattering, respectively, are large, and using 8.00 keV radiation where these corrections are smaller. The Cu atom was located by difference Patterson syntheses and used to phase the observed protein structure-factor moduli of selected reflections. An atomic model of the protein was built using a restricted data set and phases for all observed reflections to 3.0 A resolution were subsequently calculated from the initial model. The atomic model was subsequently rebuilt using all observed data. The results of this work are presented here and illustrate the utility of synchrotron radiation phasing techniques in solving the structures of metalloproteins without recourse to multiple isomorphous replacement.     

Crystal structure of a nonsymbiotic plant hemoglobin

BACKGROUND: Nonsymbiotic hemoglobins (nsHbs) form a new class of plant proteins that is distinct genetically and structurally from leghemoglobins. They are found ubiquitously in plants and are expressed in low concentrations in a variety of tissues including roots and leaves. Their function involves a biochemical response to growth under limited O(2) conditions. RESULTS: The first X-ray crystal structure of a member of this class of proteins, riceHb1, has been determined to 2.4 A resolution using a combination of phasing techniques. The active site of ferric riceHb1 differs significantly from those of traditional hemoglobins and myoglobins. The proximal and distal histidine sidechains coordinate directly to the heme iron, forming a hemichrome with spectral properties similar to those of cytochrome b(5). The crystal structure also shows that riceHb1 is a dimer with a novel interface formed by close contacts between the G helix and the region between the B and C helices of the partner subunit. CONCLUSIONS: The bis-histidyl heme coordination found in riceHb1 is unusual for a protein that binds O(2) reversibly. However, the distal His73 is rapidly displaced by ferrous ligands, and the overall O(2) affinity is ultra-high (K(D) approximately 1 nM). Our crystallographic model suggests that ligand binding occurs by an upward and outward movement of the E helix, concomitant dissociation of the distal histidine, possible repacking of the CD corner and folding of the D helix. Although the functional relevance of quaternary structure in nsHbs is unclear, the role of two conserved residues in stabilizing the dimer interface has been identified.     

Crystal structure analysis of omega-amino acid:pyruvate aminotransferase with a newly developed Weissenberg camera and an imaging plate using synchrotron radiation

The three-dimensional structure of omega-amino acid:pyruvate aminotransferase from Pseudomonas sp. F-126, an isologous alpha 4 tetramer containing pyridoxal 5'-phosphate (PLP) as a cofactor, has been determined at 2.0 A resolution. The diffraction data were collected with a newly developed Weissenberg camera with a Fuji Imaging Plate, using synchrotron radiation. The mean figure-of-merit was 0.57. The subunit is rich in secondary structure and comprises two domains. PLP is located in the large domain. The high homology in the secondary structure between this enzyme and aspartate aminotransferase strongly indicates that these two types of enzymes have evolved from a common ancestor.     

Crystal structure and ligand binding properties of the truncated hemoglobin from Geobacillus stearothermophilus

A novel truncated hemoglobin has been identified in the thermophilic bacterium Geobacillus stearothermophilus (Gs-trHb). The protein has been expressed in Escherichia coli, the 3D crystal structure (at 1.5 Angstroms resolution) and the ligand binding properties have been determined. The distal heme pocket displays an array of hydrogen bonding donors to the iron-bound ligands, including Tyr-B10 on one side of the heme pocket and Trp-G8 indole nitrogen on the opposite side. At variance with the highly similar Bacillus subtilis hemoglobin, Gs-trHb is dimeric both in the crystal and in solution and displays several unique structural properties. In the crystal cell, the iron-bound ligand is not homogeneously distributed within each distal site such that oxygen and an acetate anion can be resolved with relative occupancies of 50% each. Accordingly, equilibrium titrations of the oxygenated derivative in solution with acetate anion yield a partially saturated ferric acetate adduct. Moreover, the asymmetric unit contains two subunits and sedimentation velocity ultracentrifugation data confirm that the protein is dimeric.