Eruption patterns of permanent front teeth as an indicator of performance in roe deer

In most species of vertebrates, teeth play a central role in the long-term performance of individuals. However, patterns of tooth development have been little investigated as an indicator of animal performance. We filled this gap using data collected during long-term capture-mark-recapture monitoring of 1152 roe deer fawns at Chizé, western France. This population fluctuated greatly in size during the 27 years of monitoring, offering a unique opportunity to assess how the eruption patterns of front teeth perform as indicator of animal performance. We used three indices of the eruption of permanent front teeth, the simplest being whether or not incisor I₂ has erupted, and the most complex being a 12-level factor distinguishing the different stages of tooth eruption. We also assessed the relevance of these indices as compared to fawn body mass, a widely used indicator of animal performance of deer populations. Dental indices and body mass were positively correlated (all r > 0.62). Similarly to body mass, all indices based on tooth eruption patterns responded to changes of population size and can be reliably used to assess the relationship between roe deer and their environment. We found a linear decrease in body mass with increasing population size (r² = 0.54) and a simultaneous delay in tooth development (r² = 0.48–0.55 from the least to the most accurate indicator). However, tooth development would be not further delayed in years with the highest densities (>15 adult roe deer/100 ha). A path analysis supported the population density effect on tooth eruption patterns being mainly determined by the effect of population size on body mass. Our study provides managers with simple indices (e.g., presence-absence of I₂) that provide a technically more easy way to standardize measurements of deer density-dependent responses over large geographical and temporal scales than would be possible with body mass.     

Estimating dung decay rates of roe deer (<Emphasis Type="Italic">Capreolus capreolus</Emphasis>) in different habitat types of a Mediterranean ecosystem: an information theory approach

For elusive species living in concealing habitats (e.g. deer in a forest habitat), indirect methods such as faecal pellet counts are considered more practical means of estimating population density and abundance. Accurate estimation of deer density using the faecal standing crop (FSC) method necessitates the reliable estimation of the mean time to decay of pellet groups present during the survey. Mean time to decay is generally habitat specific, and separate estimations should be made for each habitat type in the study area. In a confined mountainous area of Greece, the habitat-specific mean time to decay of roe deer pellet groups was estimated by locating and marking fresh pellet groups on several dates in the lead up to an FSC survey and returning to the marked signs at the time of the survey to record whether or not each pellet group had survived. Several logistic models were fitted to the data, and estimations were based on a multi-model inference (MMI) approach according to information theory. The highest mean time to decay was estimated in coniferous forests, while mid-ranged values were found in maquis shrubs, and the lowest mean time to decay was observed in open areas. MMI by model averaging, based on Akaike weights, is recommended for making robust parameter estimations and for dealing with uncertainty in model selection.     

Trace mineral status and liver and blood parameters in sheep without mineral supply compared to local roe deer (Capreolus capreolus) populations

General health, clinical-chemical blood analysis and copper (Cu), zinc (Zn), selenium (Se) and vitamin E concentrations in plasma and liver tissue (wet weight, ww) of two extensive grazing sheep flocks without mineral supply were compared to the status of local roe deer (Capreolus capreolus) populations (liver samples). Both sheep flocks were classified as healthy except for a remarkable variation in body weight and a slight foot rot infection in one flock. Hematology of sheep was normal, and total protein and creatinine as well as activities of creatin kinase, aspartat-amino-transferase, alkaline phosphatase and gamma-glutamyl-transferase in plasma were within reference levels. The mean of glutamate dehydrogenase (13.8 U/l) was slightly elevated in one flock. Mean liver concentrations of Zn (38.9 and 43.5 mg/kg ww) and Cu (111 and 87.5 mg/kg ww) in sheep flocks were higher compared to the respective roe deer populations (27.5 and 36.3 mg Zn/kg ww; 18.3 and 28.6 mg Cu/kg ww). This is supposed to be caused by differences in Cu and Zn metabolism in sheep and roe deer. Selenium deficiency was diagnosed in liver samples of both sheep flocks (0.21 and 0.23 mg/kg ww). There were neither significant differences compared to roe deer (0.21 and 0.27 mg Se/kg ww) nor differences depending on location. Correlations between plasma and liver concentrations of Cu, Zn and Se were not significant in sheep. Means of vitamin E in liver samples (30.6 and 41.8 mg/kg ww) were higher in roe deer populations. This may be caused by the opportunity of selective browsing for wild ruminants, which allows access to younger plants which are higher in vitamin E.     

Population density and abundance of sympatric large carnivores in the lowland tropical evergreen forest of Indian Eastern Himalayas

Low density occurrence of large carnivore species and direct hunting of predators and prey make carnivore conservation complex. Vital baseline information on population status of large carnivores is still deficient in most forests of eastern Himalaya, which are known to be the biodiversity hotspots. To fill this information gap, we estimated the large carnivore population status and abundance in an intricate eastern Himalayan lowland tropical forest in Pakke Tiger Reserve, Arunachal Pradesh. Population status and abundance estimates of tigers and leopards were made through individual identification using closed capture-recapture sampling. To estimate the dhole abundance photographic encounter rate was used. For individually non-identifiable species photographic rate seemed to correlate well with animal abundance. The estimated tiger and leopard density through 1/2 MMDM was 2.14 ± 0.04/100 km² and 2.99 ± 1.13/100 km² respectively. Maximum likelihood estimates shows density of tiger 1.86 ± 0.7 and for leopard 2.82 ± 1.2.The estimated dhole abundance was (N) 10.6 ± 0.94, and density 6.62 ± 0.58 individuals in 100 km². Further, occupancy estimation of large carnivores may be tried along with assessing the comparative efficacy of other population estimation methods to establish better monitoring methods for this region.     

Towards a standardized index of European rabbit abundance in Iberian Mediterranean habitats

European rabbits Oryctolagus cuniculus are a keystone species in Iberian Mediterranean ecosystems. However, the reliability of methods for estimating rabbit abundance, particularly when at low numbers, is not well understood. Further, better standardization of these methodologies would allow abundance estimates to be more reliably compared between areas and periods. Consequently, we compared several frequently used methods of estimating rabbit abundance and assessed their advantages and disadvantages. During the summers of 2008 and 2009, in 11 localities of central-southern Spain we undertook (a) driving transect counts of rabbits, either at dusk or at night, (b) linear transects on foot recording rabbit signs, (c) cleared-plot pellet counts at permanent plots, and (d) standing crop counts, both with and without habitat stratification. Density estimated at night from driving transects using the Distance Sampling method (the reference method against which all other indices were compared) varied from 0 to 2.69 rabbits ha⁻¹. Most pellet-count indices were significantly related to the reference method. In particular, cleared-plot pellet counts in permanent plots corrected for pellet persistence showed the best correlation with the reference method. In contrast, latrine counts were not related to the reference method index, and we recommend against their use. A standard methodology based on cleared-plot pellets counts could be used to monitor rabbit abundance on a large scale.     

The return of the Iberian wild goat Capra pyrenaica to Portugal: From reintroduction to recolonization

The Iberian wild goat (Capra pyrenaica) is an endemic species of the Iberian Peninsula. Of the four generally accepted subspecies (Capra pyrenaica victoriae, C.p. hispanica, C.p. pyrenaica, C.p. lusitanica) only two subsist (C.p. victoriae, C p. hispanica). The subspecies once found in Portugal, C. p. lusitanica, became extinct in the 19th century. However, the reintroduction of the C. p. victoriae in north-west Spain, led to the natural recolonization of this subspecies into Portugal. Knowledge of the Iberian wild goat in Portugal is still limited, making it difficult to evaluate the major conservation/management needs. To fill this gap, here we describe the historical distribution of the Iberian wild goat in Portugal and summarize the available information on the reintroduction and recolonization process of this species. Additionally, we used line itinerary survey (2011–2012), coupled with Distance Sampling, to estimate current densities, range and population structure of the Iberian wild goat distribution in Portugal. The Iberian wild goat density is 2.78/100 ha (95% CI: 1,72–4,50; CV: 18,36%) and the abundance is of 576 (CI 95%: 356–930; CV: 18,36%). The distribution data shows that the population is divided in three nuclei: 1- Serra do Gerês (13,840 ha); 2–Serra Amarela (1235 ha); 3–Castro Laboreiro (343 ha), in a total area of 15,418 ha. Iberian wild goat population in Portugal has greatly increased in the last years, both in number and distribution range. The demographic data shows a potential for increase in the next years but our current ecological background knowledge still remains limited. Iberian wild goat management will benefit from a long-term project including public awareness, scientific research and management solutions.     

Ecological analysis of nematodes associated with vegetable-maize cropping system at Dhapa,Kolkata (India)

The study focused on the ecological analysis of plant and soil nematode communities associated with vegetables-maize crop sequences in the eastern fringe of Kolkata at Dhapa from December 2011 to April 2014 at a monthly interval. Plant-parasitic nematodes like Pratylenchus zeae, Meloidogyne incognita, Hoplolaimus indicus, Helicotylenchus dihystera, Tylenchorhynchus brevilineatus, Rotylenchulus reniformis, and Criconemoides onoensis were observed in two major cropping sequences of chilli-amaranthus-maize-radish-cauliflower and amaranthus-maize-cabbage-chilli. Among them, R. reniformis was found to be the most prevalent genera with the highest relative prominence value (RPV) of 27.29 followed by P. zeae (20.30) and T. brevilineatus (16.70). The population dynamics of P. zeae revealed the lowest density of the nematode was recorded (26 per 200 cc of soil + 5 g of the root) in March and the highest (634) in September. The variation in the abundances of other PPNs was also noted, and that attributed to the crops in the crop sequence and weather conditions. Low numbers of R. reniformis was found in the maize rhizosphere. The saprozoic nematodes (SNs) maintained higher mean densities (1734/200 cc soil); they were recorded in high numbers in the rainy season as compared to other seasons. No significant differences in the Shannon diversity index (H′), evenness (J) were observed across the growing period. A significant relationship was found between nematode abundance and climatic variables observed. This study provides baseline information on nematodes associated with maize-based cropping system, population abundance, Mylonchulus-SNs relationships, and population fluctuation in response to environmental factors.     

Crossing the border? Structure of the red deer (Cervus elaphus) population from the Bavarian–Bohemian forest ecosystem

Anthropogenic impact such as overhunting and habitat fragmentation has reduced the total red deer population (Cervus elaphus) across Europe. In Germany remaining subpopulations are even confined to designated areas with limited or no gene flow among them. Red deer populations inhabiting the Bavarian–Bohemian forest ecosystem had been divided by a fortified State border between Germany and former Czechoslovakia. To assess red deer genetic diversity more than two decades after the removal of the fortifications, we analysed a population from the Bavarian Forest National Park, Germany, and one from the National Park ?umava, Czech Republic, using 11 microsatellite loci and a 910 bp long section of the mitochondrial control region (mtDNA). Bayesian analyses of microsatellite allele frequencies favoured the presence of a single population in the Bavarian-Bohemian forest ecosystem over other population genetic structures. This admixture was supported by a lack of population pairwise differentiation between German and Czech red deer microsatellite genotypes in the analysis of molecular variance (AMOVA, F_ST = 0.009, p = 0.383). Contrastingly, AMOVA revealed a highly significant matrilinear differentiation of mtDNA between the two samples (Φ_ST = 0.285, p = 0.002), whereby German red deer belonged predominantly to haplogroup A (western Europe) and Czech red deer predominantly to haplogroup C (eastern Europe). In combination, these findings indicated a high degree of philopatry by does and extensive gene flow across the former border mediated by stags. They also identified the Bavarian–Bohemian forest ecosystem as part of a suture zone between western and eastern European red deer matrilines.     

Estimating red deer abundance in a wide range of management situations in Mediterranean habitats

When human interventions interfere with the natural regulation of wildlife populations by favouring some species, overabundance can emerge. We evaluated different methods of estimating red deer abundance in a wide range of population densities from southern Spain. Distance sampling estimates were used as the reference method across 22 localities and were compared with two kilometric abundance indices (KAIs), four indices based on pellet group counts and two browsing indices (BWIs). The average red deer density estimated by distance sampling was 19.51±3.19 deer per 100 ha, showing a wide range across the study area (0.04–66.77). Distance sampling estimates correlated with the KAIs, pellet group-based index and the BWI. The agreement with distance sampling improved when groups were used instead of individuals in the KAIs, when the minimum pellet group size was fixed at 20 pellets in the dropping counts, and when only highly palatable species were used in the BWI. Thus, several direct and indirect methods can estimate red deer abundance in Mediterranean habitats from Southern Spain with appropriate modifications.     

Habitat selection by endangered Himalayan musk deer (Moschus chrysogaster) and impacts of livestock grazing in Nepal Himalaya: Implications for conservation

Habitat management within and outside protected areas is a key to effective conservation of wildlife. This is particularly vital for declining wildlife populations within the boundary of conservation areas, while sharing their potential habitat range with foraging livestock. In an effort to understand the habitat selection by Himalayan musk deer (Moschus chrysogaster) and explore any potential impacts of livestock grazing, we conducted the present study in a conservation area of central Nepal Himalaya. We recorded data on musk deer and livestock presence and absence (based on signs of fecal pellet, footprint, and resting site) along the elevational transect with associated topographic features (elevation, slope, aspect, distance to water, and vantage point distance) and vegetation features (tree spp., shrub spp., herb spp., and canopy-cover). Using logistic regression model we found that elevation, aspect, canopy-cover, and tree spp. in the area significantly affect the likelihood of habitat selection by musk deer. In particular, they selected the southern aspect of the area with elevation ≥ 3529 m, canopy-cover ≥ 42%, and with stands of Pinus spp. and Abies spp. Slope and canopy-cover significantly affected the foraging area selection by livestock. They selected the gentler slopes in the northern aspect of the area with altitude < 3529 m and canopy-cover < 42%. Also, presence of one group of herbivore (i.e. musk deer and livestock) was not found to affect the likelihood of habitat selection by the other group. These independent habitat selections are possibly the responses to morphological and behavioral adaptations than to impacts and interactions between these two groups of herbivores. We suggest to avoid any disturbances and livestock grazing on the area that disrupt the resources and conditions likely selected and occupied by musk deer population.     

Effects of diets of differing fiber contents on digestibility,passage rate of digesta and heat production in lesser mouse deer (Tragulus javanicus)

The effects of different levels of dietary fiber on feed intake, digestibility, mean retention times of solute and particle and heat production were studied in twelve male lesser mouse deer (Tragulus javanicus). The animals were randomly assigned into four groups of three mouse deer each and fed on individual basis. Four diets, namely A, B, C and D were prepared in pellet forms. Each diet contained ～14% crude protein and ～18 kJ/g gross energy. The crude fiber contents of diet A, B, C and D were 4.2, 11.3, 16.8 and 22.5%, respectively. The results showed that the crude fiber intake of mouse deer was 1.4 ± 0.04, 3.8 ± 0.4, 5.8 ± 0.7 and 6.0 ± 0.6 g/kg W^0.75/d fed diet A, B, C and D, respectively. Mouse deer fed diet D had significantly lower DM intake than those fed lower levels of fiber. The digestibility values of DM decreased gradually with increasing levels of fiber in the diet. The mean retention times (MRTs) of particles (～1.5 mm) in the reticulorumen were in the range of (19.0–22.9 h), with the shortest time for mouse deer fed diet D. However mouse deer fed diet C and D showed significant shorter solute MRT in the reticulorumen (17.1–18.5 h) when compared to mouse deer fed lower fiber diets (21.4–21.9 h). The selectivity factor in the reticulorumen was in the range of 1.04–1.18, indicating the mouse deer to be a ‘moose-type’ ruminant. Fiber levels had no significant effect on water intake of mouse deer. Heat production tended to decrease with increasing levels of fiber (448.3–435.7 kJ/kg W^0.75/d) but differences among the mouse deer fed the four diets were not significant.     

Population densities,group size and biomass of ungulates in a lowland tropical rainforest forest of the eastern Himalayas

Large ungulate population monitoring is a crucial wildlife management tool as ungulates help in structuring and maintaining the large carnivore populations. Reliable data on population status of major ungulate prey species are still non-existent for most of the protected areas in the Indian part of the eastern Himalayan biodiversity hotspot. Twenty transects were monitored over a period of three years (2009–2011) totaling 600 km with an average length of 2 km. The estimated mean density of ungulates was 17.5 km⁻² with overall density of 48.7 km⁻². The wild pig Sus scrofa had the highest density (6.7 ± 1.2 km⁻²) among all the prey species followed by barking deer Muntiacus muntjak (3.9 ± 0.6 km⁻²), sambar Rusa unicolor (3.8 ± 0.5) and gaur Bos gaurus (3.5 ± 0.9 km⁻²). The estimated total ungulate biomass density was 2182.56 kg km⁻². This prey biomass can support up to 7.2 tigers per 100 km⁻². However, with two other sympatric carnivores sharing the same resources, the actual tiger numbers that can be supported will be lower. The estimated minor prey species was 31 km⁻² significantly 30.6% crop damages were reported by wild pig (p = 0.01) and 35.4% was elephant (p = 0.004). This data on ungulate densities and biomass will be crucial for carnivore conservation in this understudied globally significant biodiversity hotspot.     

The disappearance of muntjac (Muntiacus reevesi) and roe deer (Capreolus capreolus) pellet groups in a pine forest of lowland England

Reliable knowledge of the disappearance rate of faecal pellets is essential for converting pellet density to deer density when using standing-crop pellet-group counts. Disappearance of muntjac and roe deer pellet groups was monitored in four growth stages of a pine forest of lowland England over a 15-month period. Time to disappearance of the pellet groups (days) of both species significantly differed between habitats and months; it was shorter in late summer to early autumn and in habitats with more ground vegetation. Muntjac pellet groups disappeared more quickly than roe deer pellet groups. Time to disappearance of roe deer pellet groups was negatively correlated with air and grass temperature in pre-thicket and pre-fell habitats, while time to disappearance of muntjac pellet groups was negatively correlated with frequency of rainfall and positively correlated with the run of wind (air passage over a site within a 24-h period measured in km) in pre-thicket habitats. It is the time of the standing-crop pellet-group counts and the disappearance rate of pellet groups deposited in different months and habitats that determine the appropriate method for conversion of pellet-group density to deer density.     

At-sea abundance and spatial distribution of South American sea lion (Otaria byronia) in Chilean Northern Patagonia: How many are there?

The South American sea lion (Otaria byronia) (SSL) is a widespread opportunistic predator that inhabits waters ranging from Southern Ecuador to Southern Chile in the Pacific Ocean and from Southern Brazil to Southern Argentina in the Atlantic. SSL abundance estimates, as for many pinniped species, have relied on shore censuses, with the uncertainty that an indeterminate number of individuals at sea might remain uncounted. The proportion of the population that remains at sea during censuses and their distribution patterns are not clear and has been scarcely assessed. We used line transect sampling to gather information about at-sea abundance, density and spatial distribution of SSL in coastal waters off the Chilean Northern Patagonia. A total of 123 groups were sighted while on-effort, with an estimated density of 0.393 ind./km² (95%CI = 0.262–0.591) and a total abundance, for the surveyed area, of 13,721 individuals (95%CI = 9127–20,627). Even when our survey was rather restricted in spatial range, generalized additive model results showed that depth, distance to SSL rockeries, coastline complexity and geographical coordinates had a significant influence on SSL spatial distribution. The results on abundance and spatial distribution of SSL at sea are discussed in terms of current population estimates. Our substantial at-sea abundance estimate suggests that shore censuses might have been historically biased. Several aspects should be taken into account in further research on SSL abundance, such as including pelagic waters and undertaking simultaneously on-rockery and marine SSL abundance estimates. The results presented here provide valuable insights for revisiting and possibly improve population estimates of SSL and other pinnipeds species worldwide. This is particularly relevant when management and conservation actions need to be taken on those species that have strong interactions with fisheries and aquaculture.     

Country,Cover or Protection: What Shapes the Distribution of Red Deer and Roe Deer in the Bohemian Forest Ecosystem?

The Bohemian Forest Ecosystem encompasses various wildlife management systems. Two large, contiguous national parks (one in Germany and one in the Czech Republic) form the centre of the area, are surrounded by private hunting grounds, and hunting regulations in each country differ. Here we aimed at unravelling the influence of management-related and environmental factors on the distribution of red deer (Cervus elaphus) and roe deer (Capreolus capreolus) in this ecosystem. We used the standing crop method based on counts of pellet groups, with point counts every 100 m along 218 randomly distributed transects. Our analysis, which accounted for overdispersion as well as zero inflation and spatial autocorrelation, corroborated the view that both human management and the physical and biological environment drive ungulate distribution in mountainous areas in Central Europe. In contrast to our expectations, protection by national parks was the least important variable for red deer and the third important out of four variables for roe deer; protection negatively influenced roe deer distribution in both parks and positively influenced red deer distribution in Germany. Country was the most influential variable for both red and roe deer, with higher counts of pellet groups in the Czech Republic than in Germany. Elevation, which indicates increasing environmental harshness, was the second most important variable for both species. Forest cover was the least important variable for roe deer and the third important variable for red deer; the relationship for roe deer was positive and linear, and optimal forest cover for red deer was about 70% within a 500 m radius. Our results have direct implications for the future conservation management of deer in protected areas in Central Europe and show in particular that large non-intervention zones may not cause agglomerations of deer that could lead to conflicts along the border of protected, mountainous areas.     

Responses of wolf feeding habits after adverse climatic events in central-western Belarus

Many studies have investigated the ecology of wolf populations of Eurasia, showing that although wolves are mostly opportunistic in seeking meso-large enough mammalian prey, they can also be selective, depending on local availability of prey and their population biomass. Yet prey preferences of the wolf have been poorly evaluated in situations of complex predator/prey systems because such ecological situations are extremely rare in Europe. In particular, the role of beaver is poorly known due to the extreme decline in its range and population over the last few centuries.We conducted a 15-year study (1999–2014) of wolf Canis lupus diet in the Naliboki forest of central-western Belarus to determine the dietary responses of the wolf population in a context of a rich prey supply (beaver 650 inds/100 km², elk 47 inds/100 km², red deer 98 inds/100 km, roe deer 398 inds/100 km², wild boar 234 inds/100 km²). The bison, although present, is not preyed on. We compared the seasonal and annual diet variations of both wolf adults and pups, by scat analysis and hair identification. In winter 2012–2013, the winter was quite harsh with a long period of snow, which severely affected the roe deer and wild boar populations. Five severe summer droughts also occurred (1999, 2001, 2002, 2004 and 2013), greatly decreasing the water level in rivers and canals. We took advantage of these stressful events to evaluate the diet responses of the wolves.In “normal” years, we identified 11 food categories, essentially beaver and medium-sized ungulates (66%), and large ungulates to a lesser extent (9% in summer, 20% in winter). The adults were found to selectively supply pups with beaver, probably because of its easy transportability. Beaver consumption also increased during summer droughts when water levels were very low. After the harsh winter of 2012–2013, which was followed by a sharp decline in medium-sized prey, we observed a shift in the winter diet breadth of the wolves towards greater consumption of both large wild ungulates and small carnivores. We concluded that:1. Beaver is a functional element in wolf ecology, as a primary food for adults and pups;2. A large range of available prey species is important to maintaining a viable wolf population in cases of extreme climatic events.     

Persistent efficacy of long-acting moxidectin for control of trichostrongylid infections of sheep

The objective was to evaluate long-acting moxidectin (Cydectin^® 2% LA for sheep) against trichostrongylids in sheep. We performed a blocked, parallel, controlled clinical trial. We included 45 ewe-lambs, allocated into treated (n = 30, 1.0 mg moxidectin per kg bw, subcutaneously at base of ear on D 0) or controls (n = 15). Animals had been naturally infected (pre-treatment geometric mean epg counts: 233 and 249, respectively) and throughout the study grazed at the same pasture. Fortnightly, we collected faecal samples and performed established parasitological techniques (epg counts, coprocultures). We used analysis of covariance for post-treatment results, using pre-treatment counts as covariate, treatment as fixed effect, block as random effect. Geometric mean epg counts in treated animals were 0 up to D 112, 1 on D 126, 4 on D 140 and 13 on D 161. Respective figures for controls were 369–820 up to D 140 and 168 on D 161 (p < 0.05). Persistent efficacy (>90%) was 140 d for Teladorsagia, 119 d for Haemonchus and 115 d for Trichostrongylus. This is the first reported clinical trial of efficacy of long-acting moxidectin in sheep in Europe. Cydectin 2% LA was found to be effective against trichostrongylids of sheep. The particularly long persistent efficacy can provide new possibilities in formulating anthelmintic programs for sheep.     

Noninvasive genetic assessment of brown bear population structure in Bulgarian mountain regions

The Balkans are one of the last large refugia for brown bear (Ursus arctos) populations in Europe, and Bulgaria, in particular, contains relatively large areas of suitable brown bear habitat and a potential population of more than 600 individuals. Despite this, the majority of brown bear research remains focused on bear populations in Central and Western Europe. We provide the first assessment of genetic population structure of brown bears in Bulgaria by analysing tissue samples (n = 16) as well as samples collected with noninvasive genetic methods, including hair and faecal samples (n = 189 and n = 163, respectively). Sequence analysis of a 248 base pair fragment of the mitochondrial control region showed that two highly divergent mitochondrial European brown bear lineages form a contact zone in central Bulgaria. Furthermore, the analysis of 13 polymorphic microsatellite markers identified 136 individuals and found substantial genetic variability (H_e = 0.74; N_A = 8.9). The combination of both genetic markers revealed the presence of weak genetic substructure in the study area with considerable degrees of genetic admixture and the likely presence of migration corridors between the two subpopulation in the Rhodope Mountains and Stara Planina as evidenced from the genetic detection of two male long-distance dispersers. A detailed assessment from densely collected samples in the Rhodope Mountains resulted in a population size estimate of 315 (95% CI = 206–334) individuals, indicating that not all available habitat is presently occupied by bears in this region. Efficient management plans should focus on preserving connectivity of suitable habitats in order to maintain gene flow between the two Bulgarian brown bear subpopulations.     

Alginate embedding and subsequent sporulation of in vitro-produced Conidiobolus thromboides hyphae using a pressurised air-extrusion method

Conidiobolus thromboides is an entomophthoralean fungus with potential as a biological control agent of aphids. However, its application in biological control is limited due to its formulation requirements. The objective of this study was to develop and optimise a novel air-extrusion method to embed C. thromboides hyphae at high density in alginate pellets. An orthogonal experimental design was used to investigate selected combinations of parameters known to affect hyphal density within pellets. The diameter of pellets produced, and the calculated density of hyphae within them, ranged from 0.18 ± 0.09 to 3.17 ± 0.06 mm and from 0.02 to 350.56 mg/mm³ respectively. These data were used to predict the optimal parameter combination to deliver the greatest density of hyphae of C. thromboides per pellet: 1% sodium alginate, a 1:2 ratio of hyphae to sodium alginate, an orifice diameter of 0.232 mm and an air pressure of 0.05 MPa. Pellets made under the optimal conditions predicted produced a mean total of 4.3 ± 0.6 × 10⁵ conidia per pellet at 100% relative humidity which was significantly greater than the mean total number of conidia produced from infected aphid cadavers of comparable size (9.35 ± 0.85 × 10⁴) (p < 0.001). In conclusion, air-extrusion embedding appears to be a promising method for formulating in vitro-produced hyphae of C. thromboides for use in biological control.     

Plasma gossypol dynamics in white-tailed deer: Implications for whole cottonseed as a supplemental feed

Whole cottonseed (WCS) is a potential supplemental feed for white-tailed deer (Odocoileus virginianus) in rangeland conditions because of its high digestible energy and protein content, moderate fiber content, and resistance to degradation in moist conditions. WCS also contains the polyphenolic secondary metabolite gossypol, which reduces palatability to non-target monogastric species but may be of concern for deer nutrition. Plasma gossypol stabilization when fed a constant dry matter intake, plasma gossypol depletion after WCS was removed from the diet, and the relationship between WCS consumption and plasma gossypol concentration was studied in 10 mature male (N = 5) and female (N = 5) captive white-tailed deer. Consumption of WCS by 73 free-ranging white-tailed deer (59 males, 14 females) was estimated using results of the captive study. Plasma gossypol concentrations declined exponentially and averaged 0.74 μg/mL 35 days after WCS was removed from the diet. Plasma gossypol concentration was linearly related to WCS consumption (P < 0.001), with females having 0.35 μg/mL greater (P = 0.04) plasma gossypol than males for a given rate of dry matter consumption. All female and 93% of male white-tailed deer captured in WCS supplemented pastures had detectable plasma gossypol. Female averaged 1.88 μg/mL of plasma gossypol and males averaged 4.84 μg/mL of plasma gossypol. Based on the captive deer data, these plasma values suggest an average WCS consumption of ∼2.6 g/kg BW/day for female free-ranging deer and ∼5.6 g/kg BW/day for male free-ranging deer. Inferentially, a large proportion of free-ranging white-tailed deer in rangeland conditions will consume WCS, with females consuming 125 g WCS/day and males consuming 428 g WCS/day. That plasma gossypol levels decrease rapidly after cottonseed is removed from the diet suggests that the long withdrawal periods often used prior to breeding season may not be needed. However, although 93% of gossypol was eliminated from the animals after a five-week withdrawal period, a small amount of gossypol can still be detected. While our preliminary data on these animals suggests that these levels are not detriment to animal health or reproduction, ranch managers may want to take a conservative approach to the feeding of WCS until these questions are answered.