Identification of Mycobacterium avium subsp. paratuberculosis in wild cervids ( Cervus elaphus hippelaphus and Capreolus capreolus ) from Northwestern Italy

Seventy-seven red deer (Cervus elaphus hippelaphus), 40 roe deer (Capreolus capreolus) from the Northwestern (NW) Alps (Turin Province, NW Italy) and 29 roe deer from the NW Apennines (Alessandria province, NW Italy) were examined for the presence of Mycobacterium avium subsp. paratuberculosis (MAP) by culture, IS900 nested polymerase chain reaction (PCR) and IS1311 PCR restriction endonuclease analysis for strain characterisation. MAP identification (nested PCR and/or culture) allowed us to detect 32.9% MAP-infected red deer and 22.5% infected roe deer in the NW Alps and 41.4% MAP infected roe deer in the NW Apennines. On the basis of the polymorphism present in the IS1311 sequence, all MAP isolates were characterised as cattle strains. Our results show that MAP circulates widely among populations of wild cervids in NW Italy.     

Selenium concentration in liver and kidney of free living animals (roe and red deer) from West Pomerania (Poland)

The aim of this study was to determine concentrations of selenium in the liver and kidneys of roe deer and red deer from West Pomerania, depending on the season. Altogether, samples from 169 animals were collected (96 from roe deer and 73 from red deer) in 2003–2007. The mean concentration of selenium in the liver of red deer and roe deer was 0.37 μg/g and 0.62 μg/g dry weight, respectively. In kidneys, Se concentration was 2.72 μg/g d.w. in red deer and 2.99 μg/g d.w. in roe deer. In roe deer, liver selenium concentration in autumn was significantly higher than in winter (P < 0.05) and spring (P < 0.01) and significantly lower in spring than in summer (P < 0.05); likewise, kidney selenium concentration was higher in autumn than in summer. In deer, no statistically significant season-related differences were observed for liver selenium concentrations. In red deer kidneys, selenium concentration was the lowest in summer, significantly lower than in autumn and winter. Low selenium concentrations in the analyzed tissues show that the animals live in areas deficient in this element.     

Detection of Bartonella DNA in roe deer (Capreolus capreolus) and in ticks removed from deer

The potential role of roe deer as a sylvatic reservoir of Bartonella in north-west Poland has been assessed. In addition, ticks infesting roe deer were screened to assess their role as a vector and reservoir of Bartonella. Blood and tissue samples of 72 animals from north-western Poland were PCR-screened. Bartonella DNA was detected by using primers complementary to the intergenic spacer between the 16S and 23S rRNA genes, which is used for identification of over a dozen species of this genus. Products of three different sizes were detected: 230 and 290 bp, representative of two strains of Bartonella capreoli, and 190 bp, identified as Bartonella bovis. All the three amplicons were detected in the blood, spleen and liver from the roe deer. All samples from the heart, lungs and kidneys were PCR negative. In ticks (Ixodes ricinus), only the 290 bp fragment from B. capreoli was present. Generally, Bartonella infection rate in Capreolus capreolus amounted to 27.6% of the roe deer, but it was much higher during winter (62%) than in spring (6.9%). The results show that the roe deer may be a reservoir for B. capreoli and B. bovis. The infection detected in I. ricinus ticks (7.7%) suggests that ticks may act as a Bartonella reservoir and vector.     

Seasonal dynamics of digestion in relation to diet quality and intake in European roe deer (Capreolus capreolus)

Three tame, hand-reared adult roe deer (Capreolus capreolus) were fed a low-quality diet (blueberry stems) or high-quality concentrate either restricted or ad libitum in summer and winter. Total mean retention time (TMRT) and rumen retention time (RRT) were significantly longer, indigestible fill was significantly higher and apparent digestibility and intake were significantly lower when fed blueberry stems compared to concentrate. The feeding regimes of concentrate did not affect the digesta kinetic estimates, except that total fill and apprent digestibility were significantly higher when deer were fed ad libitum than when they were fed a restricted ration. TMRT was significantly longer when they were fed a restricted ration of blueberry stems than for ad libitum intake, whereas all other estimates were equal. Intake was significantly higher and TMRT and RRT were significantly shorter in summer compared to winter when they were fed concentrate. These findings support the theory that roe deer, as small-bodied concentrate selectors, are adapted to high intake, rapid turnover, and rapid digestion when fed high-quality concentrate. When fed a low-quality diet the roe deer may be limited by rumen bulk. A strategy based on increased rumen retention accompanied by increased cell-wall digestion has therefore severe limitations in roe deer. This indicates that roe deer are dependent on a selective feeding strategy to enhance winter survival.     

Comparative evaluation of effort, capture and handling effects of drive nets to capture roe deer (Capreolus capreolus), Southern chamois (Rupicapra pyrenaica) and Spanish ibex (Capra pyrenaica)

The objective of this study is to assess the usefulness of drive nets to capture roe deer (Capreolus capreolus), Southern chamois (Rupicapra pyrenaica) and Spanish ibex (Capra pyrenaica), comparing the results obtained with other capture methods and amongst the three species. Sixty-five drive net capture operations using beaters were conducted from January 1998 to September 2004. A total of 161 wild ungulates (31 roe deer, 95 Southern chamois and 35 Spanish ibexes) were captured. The average number of animals captured per operation was 1.07 for roe deer, 3.96 for Southern chamois and 2.92 for Spanish ibex. The average number of person–days per captured animal was 21.5, 7.1 and 10.6 for roe deer, Southern chamois and Spanish ibex, respectively. Specificity was 100% for Southern chamois and Spanish ibex (only the target species captured) and 77.5% for roe deer. Risk for the animals (mortality plus injuries) was 3.23% for roe deer, 5.27% for Southern chamois and 0% for Spanish ibex, whereas injuries to the operators occurred with 3.1% of the handled animals. Sex ratio was skewed towards females in roe deer, towards males in Southern chamois and balanced in Southern chamois. Drive nets showed good performance, although many operators were required. Safety for the animals and specificity were higher than traditionally attributed to this capture method. It is concluded that drive nets are an efficient and safe method to capture many ungulate species.     

Ranging behaviour and excursions of female roe deer during the rut

Anecdotal evidence has suggested that, during the rutting period, female roe deer may undertake short excursions, outside of their normal home range, possibly to mate with a reproductive partner. To address this question, we analysed the ranging behaviour of 27 female roe deer Capreolus capreolus, equipped with GPS collars, inhabiting a fragmented landscape in France. We compared female movements during the rutting period with a non-rutting period over two summers using a recently published approach. Search intensity and home range size were significantly greater during the rutting period. The difference in home range size between the two periods was significantly greater in 2006 compared to 2005 and in open compared to closed habitat. We were not able to identify any influence of body mass on the difference in ranging behaviour between the two periods. Visual analysis of movement trajectories for 11 females revealed that 5 (45%) performed an excursion for a duration of a few hours to several days. We speculatively suggest that female rut excursions provide an opportunity for active mate choice in roe deer, where males are territorial, although we cannot rule out the alternative explanation that these movements are a means to avoid male harassment.     

Analysis of parotid and mixed saliva in Roe deer (Capreolus capreolus L.)

In ruminants, different functions have been ascribed to the different salivary glands according to the feeding type. In this context, possible adaptations of salivary functions were investigated regarding the secretion of various proteins by different types of salivary glands. To yield uncontaminated parotid saliva in large quantities, a non-surgical method has been developed. Parotid gland secretions were collected via endoscopic placement of guide wires into each parotid duct, which were subsequently used for placement of collection catheters. Salivary flow was stimulated by intra-glandular administration of the parasympathomimetic compound pilocarpine-hydrochloride into the parotid gland. Mixed saliva (excluding parotid saliva) was collected into sterile tubes by normal outflow during the sampling of parotid saliva. The total flow volume, flow rate and the content of proteins as well as of several ions (Na⁺, K⁺, Ca²⁺, inorganic phosphate) of both types of saliva were measured in sheep, fallow deer and roe deer. Roe deer secreted the highest amount of total salivary proteins relative to body mass [mg/kg body mass] and the highest relative volume [ml/10 min/kg body mass], both in parotid and mixed saliva, of all ruminant species examined. Additionally, the protein profile and the tannin-binding properties of parotid and mixed saliva in roe deer were investigated. Parotid saliva bound almost twice as much tannin as mixed saliva, underlining the importance of yielding uncontaminated parotid saliva for tannin-binding studies. Accepted: 6 January 1998     

Isolation of cellulose-hydrolytic bacteria and applications of the cellulolytic enzymes for cellulosic biohydrogen production

Nine cellulolytic bacterial strains were isolated from soil sample taken in southern Taiwan. Through 16S rRNA sequence matching; eight of those isolates belong to Cellulomonas sp., while the other one belongs to Cellulosimicrobium cellulans. The activity of cellulolytic enzymes (cellulases and xylanase) produced from those strains was mainly present extracellularly and the enzyme production was dependent on cellulosic substrates (xylan, rice husk and rice straw) used for growth. HPLC analysis confirmed the bacterial hydrolysis of these cellulosic substrates for soluble sugars production. The efficiency of fermentative H₂ production from the enzymatically hydrolyzed rice husk was examined with seven H₂-producing pure bacterial isolates. With an initial reducing sugar concentration of 0.36 g l⁻¹, only Clostridium butyricum CGS5 exhibited efficient H₂ production from the rice husk hydrolysates with a cumulative H₂ production and H₂ yield of 88.1 ml l⁻¹ and 19.15 mmol H₂ (g reducing sugar)⁻¹ (or 17.24 mmol H₂ (g cellulose)⁻¹), respectively.     

Temporal shifts of DNA‐microsatellite allele profiles in roe deer (Capreolus capreolus L.) within three decades

DNA‐microsatellite polymorphism (four loci) was studied in 56 male roe deer (Capreolus capreolus) from a 900‐ha hunting territory in the Vosges du Nord Mountains (France), culled over 34 years (1956–1990). Changed allele frequencies at two loci within this period, and increased allelic diversity, were traced to a phase of reduced population density and subsequent immigration. Decadic population samples collected within 900‐ha were distinguished by higher genetic variability measures than were certain geographical samples across Central Europe (4–900 km). On average, the decadic cohorts were distinguished by a gene diversity index of G_ST = 0.0286, and a genetic distance of D = 0.0938, which reflect 54% (G_ST) and 69% (D) of the respective geographic (350 km) differentiation indices of roe deer in Central Europe. The importance of demography and population ecology effects for microevolution in a large mammal is demonstrated, as is the risk of artefact by composing population samples of deer over several years. Population genetic screening should cover various demes of roe deer from the same general region, and be based on many unlinked polymorphic loci, to minimize the distorting effects of genetic dynamics at the small spatial scale.     

Description and epidemiological implications of <Emphasis Type="Italic">S. scabiei</Emphasis> infection in roe deer (<Emphasis Type="Italic">Capreolus capreolus</Emphasis>) originating from chamois (<Emphasis Type="Italic">Rupicapra rupicapra</Emphasis>)

The authors describe a case of Sarcoptes scabiei transmission from a chamois (Rupicapra rupicapra) kid to three roe deer (Capreolus capreolus) fawns kept in captive cohabitation in a wildlife rehabilitation centre for approximately 4 months. Symptoms in the two male fawns started developing 1 week after skin lesions were noticed in the kid, whereas a delayed (75 days later) appearance of scabies signs was observed in the female fawn. A severe generalised form of scabies rapidly developed in a male fawn, but lesions in the other two healed after triple inoculation of ivermectin at 15 days intervals. These roe deer were monitored for specific anti-S. scabiei antibodies with an immunoblotting technique. The strongest antibody response was exhibited during the clinical phase of the infection and then rapidly declined post-treatment. Interestingly, the female fawn developed immunoglobulin G to S. scabiei proteins more than 2 months before the onset of clinical scabies. Since all captive roe deer appeared sensitive to infection with S. scabiei var. rupicaprae, the authors hypothesise that the sporadic frequency of spontaneous scabies in this Cervid host may be attributed to avoidance of direct contacts with moribund or recently dead scabietic chamois, and/or to a self-limiting (and therefore difficult to detect) course of the disease after infrequent infectious contacts and/or exposure to lower infectious doses than in the described episode.     

Apoptosis is not the cause of seasonal testicular involution in roe deer

Apoptosis is involved in the regulation of spermatogenesis. The involution of testes in seasonal breeders might be expected to involve enhanced apoptotic cell elimination. We have compared seasonally changing testicular apoptosis in roe deer with that in non-seasonally breeding cattle. Apoptotic cells were detected as TUNEL-positive cells by both flow-cytometric analysis and in situ localisation of fragmented DNA in tissue sections. Apoptosis-induced DNA fragments were also assessed by enzyme-linked immunosorbent assay (ELISA) in homogenised testicular parenchyma. As expected, the testis mass and the percentage of haploid cells in roe deer showed a seasonal pattern with a significant maximum during the rut (August), whereas no annual variation of these parameters was found in bulls. All three methods for determining apoptosis showed similar findings. Roe deer exhibited significant seasonal fluctuation of total apoptotic activity (ELISA, apoptotic cells per tubule cross section) with a maximum during the breeding season. However, the seasonal differences in the number of apoptotic cells corresponded to the variable total numbers of spermatogonia and spermatocytes per tubule cross section. Thus, the percentages of TUNEL-positive cells related to the combined number of both germ cell types showed no seasonal variance, as confirmed by percentages of apoptotic cells analysed flow-cytometrically. The maximum level of apoptosis during the rut in roe deer was similar to the values obtained during the invariably high spermatogenic activity in cattle. These results suggest that, in roe deer, apoptosis is not the cause of the seasonal involution of testes. This study was partially supported by grant Bl 319/6-1 from the Deutsche Forschungsgemeinschaft.     

Minimal activity in both proliferation and apoptosis of interstitial cells indicates seasonally persisting Leydig cell population in roe deer

Seasonally regulated breeding is associated with significant changes in testis mass, structure and function. This includes the variation in size, structure and function of the Leydig cells. Recently, interstitial cells have been characterised as a numerically constant population in roe deer. However, no consistent data are available regarding changes in the number of Leydig cells, their differentiation or turnover in seasonally breeding mammals. This study has quantified the numbers of both proliferating and apoptotic cells in roe deer testis bimonthly during a complete annual cycle. Proliferation was detected by immunolocalisation of PCNA and Ki-67 in tissue sections, whereas apoptosis was localised by the TUNEL technique and an antibody to caspase-3. The labelled cells were counted by using a computer-aided image-analysing system. The number of proliferating spermatogenic cells per tubule cross section showed seasonal changes with a maximum in April (14.9±0.6) and a subsequent decline up to December (1.6±0.3). Percentages of positive cells per square millimetre of interstitial area were below 1% throughout the year. The average number of apoptotic cells per tubule cross section was low and varied only between 0.5 and 1.4 (caspase-3) or 0.1 and 2.1 (TUNEL). In the interstitial compartment, only a few apoptotic cells (0.7%) were found sporadically scattered within the intertubular region during all studied seasonal periods. The results suggest that a constant total number of interstitial cells arise from a conserved cell population of changing functional state rather than from a steady-state population with a definite turnover of cells during seasonal changes in testicular activity.The study was supported by a grant from the Deutsche Forschungsgemeinschaft (BL 319/6-2).     

四川梅花鹿春季昼夜活动节律与时间分配

1997年4月,在四川铁布自然保护区用直接观察法对220头次四川梅花鹿群的昼夜活动节律和时间分配进行了观察。结果表明,鹿群春季昼夜活动的规律性较强。白昼,鹿群活动呈现明显的双峰型,2个高峰时段为08：30和19：30前后,活动频率分别为71．9％和94．13％,其他时间多处于休息状态;鹿群夜间活动强于昼间,活动频率均在6l％以上,仅在半夜01：30前后有一个相对不活跃期。同时发现,春季活动高峰期约90％的活动时间被四川梅花鹿用于采食和移动。这可能与光照、食物及人类活动的影响有关。     

Fighting behaviour in territorial male roe deer <Emphasis Type=Italic>Capreolus capreolus</Emphasis>: the effects of antler size and residence

In all areas where they have been studied, male roe deer are believed to have a territorial mating system, although few quantitative studies have been conducted and there remains considerable debate about the function of male roe deer territories. We observed 139 aggressive interactions between male roe deer in Storfosna Island (Norway) during one territorial season (March–August). We recognised seven rank levels of escalation according to the potential danger of the behaviour. On the basis of the number of escalation levels included in the interactions, the complexity of the fights was also scored. We recorded the presence of other individuals during the interaction, the age, the antler size, the territorial status and the residency status of the two contestants and tested how these variables affected escalation, complexity and outcome of the fights. Most of the interactions ended with low levels of escalation, and physical contact occurred only in fights between two territorial bucks. The escalation was also affected by the difference in antler size index (the bucks escalated more when the difference in antler size was smaller) and increased with an increasing number of female deer present during the interaction. The resident buck won in 81% of the fights. When it drew or lost, it was generally both inferior in age and antler size, and the duration and escalation of the interactions were higher. However, even when a fight was lost, no territory loss occurred. These results are consistent with the evolutionary game theory and the proposed low risk–low gain strategy of roe deer bucks.     

Concentrations of insulin-like growth factor-I in adult male white-tailed deer (Odocoileus virginianus): associations with serum testosterone, morphometrics and age during and after the breeding season

Our understanding of insulin-like growth factor-I (IGF-I) in cervids has been limited mostly to its effects on antler development in red deer (Cervus elaphus), roe deer (Capreolus capreolus), fallow deer (Dama dama), and pudu (Pudu puda). Although IGF-I has been found to play a critical role in reproductive function of other mammals, its role in reproduction of deer is unknown. The objectives of the present study were to determine if serum levels of IGF-I change during the breeding season, assess whether age influences serum IGF-I, compare levels of IGF-I measured during and following the breeding season, and determine if IGF-I is associated with body and antler characteristics in free-ranging adult, male white-tailed deer (Odocoileus virginianus). We collected serum and morphometric data from hunter-harvested and captured white-tailed deer to investigate these objectives. Mean level of serum IGF-I during the breeding season was 63.6 ng/ml and was greatest in deer between 2.5 and 5.5 years old (57.4-79.9 ng/ml). Levels of serum IGF-I decreased by approximately 40% as the breeding season progressed, but levels were less in deer following the breeding season (34.6 ng/ml). Both body and antler size were associated positively with IGF-I when controlling for age. Serum testosterone was also associated positively with IGF-I. Levels of serum testosterone during the breeding season generally increased with age from 4.82 (1.5 years old) to 18.79 ng/dl (5.5 years old), but decreased thereafter. These data suggest that IGF-I may be an important hormone in breeding, male white-tailed deer.     

Sucking and allosucking duration in farmed red deer (Cervus elaphus)

Sucking duration in ungulates does not only mean milk transfer, but is also associated with maternal care in general. It seems to be a reflection of offspring demand rather than solely milk transfer rate. Thus, the objective of this study was to discriminate between sucking and allosucking (i.e. sucking non-maternal hind) behaviour in red deer according to the sucking duration.We hypothesized that: (1) calves should suck longer from their mothers than allosuck from non-maternal hinds; (2) sucking duration of calves frequently nursed by a particular non-maternal hind should be longer than that of calves occasionally allonursed; (3) sucking duration should be longer for bouts including one calf than two or more calves sucking simultaneously; (4) male calves should suck and allosuck longer than female calves; and (5) primiparous hinds should nurse and allonurse longer than multiparous hinds. We observed sucking behaviour of 25 hinds and their 38 calves (from birth until the youngest calf reached one month of age) in two seasons. We recorded 1730 sucking bouts, of which 11.62% in the first season and 4.37% in the second season were non-filial. The duration of filial sucking was significantly longer than non-filial sucking. A large individual variance in the incidence of non-filial sucking in both the calves and hinds was found. Therefore, the non-filial hind–calf pairs were categorized in two clusters according to the frequency of nursing non-filial calves for one hind in relationship to all nursing events for this hind by a cluster analysis (PROC CLUSTER, SAS). We used a general linear mixed model, GLMM (PROC MIXED, SAS) to test the influence of hind relationship to the nursed calf (filial, frequently allosucking non-filial, or occasionally allosucking non-filial pair). Sucking duration of occasionally allosucking non-filial calves was only marginally different from that of filial calves. There was no difference between the two groups of non-filial calves. Multiple sucking bouts were shorter than those with one calf. Male calves sucked longer than female calves; however, the greatest difference was recorded between frequently allosucking non-filial pairs of both sexes. Frequently allosucking non-filial males sucked the longest and differently from occasionally allosucking non-filial males. Frequently allosucking non-filial females sucked the shortest and differently from filial calves of both sexes. It is more likely that allosucking seems to be more important for male rather than female calves.Therefore, it is concluded that allosucking calves differ in their sucking behaviour and two types of allosuckers (frequent and occasional) should be taken into account when analyzing allosuckling behaviour.