scholarly journals 190 A PROCEDURE TO OBTAIN FIBROBLASTS FROM WILD ANIMALS

2005 ◽  
Vol 17 (2) ◽  
pp. 245 ◽  
Author(s):  
J.N. Caamaño ◽  
M. Hermsen ◽  
J. Marcos ◽  
A. Meana ◽  
C. Alonso ◽  
...  

The number of wild animals threatened or at risk of extinction constantly increases and, as a result, biodiversity decreases. Therefore, it is necessary to set in motion proceedings to preserve the genetic variability and to overcome factors leading to this situation. Banks of cryopreserved cells, such as skin fibroblasts, from wild animals could be used both for genetic studies and as a cell source for technologies able to reproduce individuals. In this study we report that a single basic protocol provides a method for obtaining, culturing, and cryopreserving skin fibroblasts from a wide range of wild animals. Skin biopsies were taken in Cantabria (Spain) from live, anesthetized brown bear (Ursus arctos, n = 4), and in Asturias (Spain) from carcasses of accidentally killed individuals from the following species: grey wolf (Canis lupus, n = 1), red fox (Vulpes vulpes, n = 1), eurasian badger (Meles meles, n = 1), ermine (Mustela erminea, n = 1), fallow deer (Dama dama, n = 7), pyrenean chamois (Rupicapra pyrenaica, n = 1), western roe deer (Capreolus capreolus, n = 1), wild boar (Sus scrofa, n = 1), striped dolphin (Stenella coeruleoalba, n = 1) and one avian species, capercaillie (Tetrao urogallus, n = 1). Once obtained and transported to the laboratory, all samples were processed identically. Tissue samples were manually cut into small pieces and enzymatically digested with 0.05% trypsin-EDTA (Sigma-Aldrich, Madrid, Spain) for 30 min, and then with collagenase (300 units/mL) (Sigma) for 14 h at 38°C. Disaggregated cells were centrifuged at 1600 rpm for 10 min and the pellet was diluted with D-MEM (Sigma) containing 10% FBS. Cells were counted and a minimum of 3 × 105 cells were placed in a 25 cm2 flask for culture under 5% CO2 in air and high humidity at 37°C (red fox, badger, ermine, western roe deer) or 38°C (brown bear, grey wolf, fallow deer, pyrenean chamois, wild boar, striped dolphin, capercaillie). Confluent fibroblast monolayers were obtained after 5–7 days in culture except for the striped dolphin whose sample was contaminated. Two to three passages were performed using 75 cm2 flasks before freezing fibroblasts in D-MEM containing 10% DMSO (Sigma) and 10% FBS. Total cell yield from a confluent flask was 5–11 × 106 cells, depending on the animal species. Fibroblasts recovered from all thawed samples grew to confluence, and karyotypes were performed at this step. It was possible to obtain, culture, freeze, and thaw skin fibroblasts successfully from all the above-mentioned land species. Although more research is needed, preliminary data suggest that the above-described procedure can be applied to mammals and birds, thus helping to preserve genetic material of wild animals. Grant Support: Consejería de Medio Ambiente, Ordenación del Territorios e Infraestructuras, Asturias, SPAIN. The authors acknowledge CEPESMA, Luarca, Asturias, SPAIN.

2021 ◽  
Vol 75 (2) ◽  
Author(s):  
Mariana Rossa ◽  
Sandro Lovari ◽  
Francesco Ferretti

Abstract Spatial and temporal occurrence can mediate behavioural interactions between apex predators, mesocarnivores and herbivores. Predators should adapt their activity to that of prey, whereas predator avoidance would be expected to influence activity patterns and space use of prey and smaller competitors. We evaluated interspecific spatiotemporal relationships in a prey-rich community including an apex predator (the wolf), three wild ungulates and several smaller herbivores/mesocarnivores, through camera trapping. All considered species (i.e. wolves and potential prey/smaller competitors: wild boar, fallow deer, roe deer, crested porcupine, red fox and European badger) were active especially at night and/or twilight. Among wild ungulates, the wolf showed the greatest temporal overlap with the wild boar and the lowest one with the least abundant and used of them, i.e. the roe deer. The main prey (i.e. the fallow deer) showed more diurnal activity and a lower temporal overlap with the predator in sites with high wolf activity than in low-activity ones. Among mesocarnivores, the red fox showed extensive temporal overlap with the wolf: the overlap between the two canids was greater in sites intensively used by this apex predator than in sites with low wolf activity, supporting a concurrent study which suggested a potential for facilitative—rather than competitive—interactions. Spatiotemporal relationships suggest complex interactions between the apex predator, prey and smaller carnivores, for which a substantial temporal or spatial association was often supported. Significance statement There is a growing interest in the influence of apex predators on ecosystems through their effects on the behaviour of prey and smaller carnivores, especially in the light of the ongoing recovery of large carnivores in temperate areas. Predators should synchronise their activity to that of prey; conversely, prey and smaller carnivores would be expected to avoid predators. In a rich community including the wolf, three wild ungulates and several mesomammals, we detected (i) a substantial temporal overlap between wolves and wild boar, porcupines and mesocarnivores; (ii) a negative temporal association between the predator and its main prey (i.e. the fallow deer) and (iii) a great temporal overlap between the wolf and the red fox. We provide a baseline to evaluate temporal changes of predator-prey-mesocarnivore behavioural interactions along with variations of carnivore-prey densities.


2020 ◽  
Vol 41 (2) ◽  
pp. 347-358 ◽  
Author(s):  
Joso Vrkljan ◽  
Dubravka Hozjan ◽  
Danijela Barić ◽  
Damir Ugarković ◽  
Krešimir Krapinec

The purpose of this study was to determine the frequency of wildlife-vehicle collisions (WVC) based on the animal species, and to deepen the knowledge of temporal patterns of vehicle collisions with roe deer and wild boar. The study analyses the data from police reports on vehicle collisions with animals on state roads, by date and time, section of road, and animal species over a 5-year period (2012–2016). These data were analysed to determine the temporal dynamics of vehicle collisions with roe deer and wild boar by month, time of day, and moon phase. On the state roads in the Dinaric area, roe deer are most commonly involved in vehicle collisions (70.1% of all collisions), followed by wild boar (11.0%). Other large species involved in collisions were fallow deer (4.8%), brown bear (1.8%), red deer (0.9%), grey wolf (0.7%), and European mouflon (0.5%), respectively. Most collisions with roe deer occurred in the period April–August, with reduced frequency during autumn and winter. For wild boar, there was no association between month and frequency of collisions. At the annual level, collisions with roe deer were significantly higher during night (37%) and twilight (41%) than during the day (22%). For wild boar, most collisions occurred during twilight (26%) and night (72%), although the difference between these two periods was not statistically significant. For roe deer, collisions had no association with lunar phase, though wild boar collisions during twilight (dawn or dusk) were more common during twilight periods on days with less moonlight. Since vehicle collisions with wildlife showed certain temporal patterns, these should be taken into consideration in developing statistical models of spatial WVC patterns, and also in planning strategies and countermeasures to mitigate WVC issues.


Animals ◽  
2020 ◽  
Vol 10 (6) ◽  
pp. 1014
Author(s):  
Sayantani M. Basak ◽  
Izabela A. Wierzbowska ◽  
Agnieszka Gajda ◽  
Marcin Czarnoleski ◽  
Maciej Lesiak ◽  
...  

Efforts to reduce human-wildlife-conflict are integral to wildlife management and conservation in urban habitats. In our study, we identified the HWC situations in urban areas of Krakow city, based on animal-vehicle collisions, intrusion to property, and damages. Hot spot analysis and Moran’s Index were used to identify the location of maximum potential conflict. We analysed 2512 incidents in which animals (of which 85% included mammals and 15% birds) were involved in conflict situations between 2007 and 2013. A significant seasonal variation was observed among the animals. We also identified roe deer (50.23%), red fox (22.80%) and wild boar (11.40%), as the three prominent conflicted animals. Getis–Ord Gi* analysis was used to identify spatial clusters of conflict. A significant spatial association was found in the location of clusters of hot spots in specific land-use based on Moran’s Index. Hot spots of roe deer and wild boar were high in grasslands and in forest and for red fox in built-up area. The results underscore the notion that conservation and wildlife management efforts must take into account differences in the seasonality of HWC among species. This information can be used to inform mitigation strategies.


2019 ◽  
Vol 88 (1) ◽  
pp. 103-111 ◽  
Author(s):  
Michaela Nesvadbová ◽  
Gabriela Bořilová ◽  
Radka Hulánková

Meat authentication is currently a key topic in relation to the quality and safety of food of animal origin at all levels of production and the global distribution chain. New polymerase chain reaction-restriction fragment length polymorphisms (PCR-RFLP) based on digestion of PCR products with two restriction enzymes, MboII and AciI, have been developed for the specific identification of raw and heat-processed meat from red deer (Cervus elaphus), sika deer (Cervus nippon), roe deer (Capreolus capreolus), fallow deer (Dama dama), mouflon (Ovis musimon), wild boar (Sus scrofa), hare (Lepus europaeus) and cattle (Bos taurus). The PCR primers were targeted in a well-conserved region of the cytochrome b (CYTB) gene to amplify a 378 bp region of all the analysed species. This simple, rapid and cost-effective method is suitable for identification of the meat of game species and their possible substitution by beef.


Animals ◽  
2021 ◽  
Vol 11 (9) ◽  
pp. 2722
Author(s):  
Diana Žele-Vengušt ◽  
Renata Lindtner-Knific ◽  
Nina Mlakar-Hrženjak ◽  
Klemen Jerina ◽  
Gorazd Vengušt

A total of 249 serum samples from 13 wild animal species namely fallow deer (Dama dama, n = 1), roe deer (Capreolus capreolus, n = 80), red deer (Cervus elaphus, n = 22), chamois (Rupicapra rupicapra, n = 21), mouflon (Ovis musimon, n = 4), brown hare (Lepus europaeus, n = 2), nutria (Myocastor coypus, n = 1), red fox (Vulpes vulpes, n = 97), stone marten (Martes foina, n = 12), European badger (Meles meles, n = 2), golden jackal (Canis aureus, n = 2) Eurasian lynx (Lynx lynx, n = 2) and grey wolf (Canis lupus, n = 3) were analysed for the presence of antibodies against Leptospira interrogans sensu stricto. Serum samples were examined via the microscopic agglutination test for the presence of specific antibodies against Leptospira serovars Icterohaemorrhagiae, Bratislava, Pomona, Grippotyphosa, Hardjo, Sejroe, Australis, Autumnalis, Canicola, Saxkoebing and Tarassovi. Antibodies to at least one of the pathogenic serovars were detected in 77 (30.9%; CI = 25–37%) sera. The proportion of positive samples varied intraspecifically and was the biggest in large carnivores (lynx, wolf and jackal; 86%), followed by mezzo predators: stone marten (67%) and red fox (34%), and large herbivores: red deer (32%), roe deer (25%), alpine chamois (10%) and mouflon (0%). Out of the 77 positive samples, 42 samples (53.8%) had positive titres against a single serovar, while 35 (45.4%) samples had positive titres against two or more serovars. The most frequently detected antibodies were those against the serovar Icterohaemorrhagiae. The present study confirmed the presence of multiple pathogenic serovars in wildlife throughout Slovenia. It can be concluded that wild animals are reservoirs for at least some of the leptospiral serovars and are a potential source of leptospirosis for other wild and domestic animals, as well as for humans.


2020 ◽  
Author(s):  
Sorour Karimi ◽  
Mahmoud-Reza Hemami ◽  
Mostafa Tarkesh Esfahani ◽  
Christophe Baltzinger

Abstract Background: Vertebrate-mediated seed dispersal is probably the main long distance dispersal mode. Through endozoochory, large mammals act as mobile links between habitats within and among forest patches. Along with other factors, their feeding regimes do affect their contribution as dispersal vectors. We conducted a cross-species comparative experiment involving two herbivores, red deer and roe deer; and two opportunistic omnivores, wild boar and brown bear, all occurring in the forest and steppe-forest ecotone habitats of the south-eastern Caspian region. We compared their role as endozoochorous seed dispersal agents by monitoring seedling emergence in their dungs under greenhouse and natural conditions. Results: In total, 3078 seedlings, corresponding to 136 plant taxa sprouted from 445 paired dung sub-samples, under greenhouse and natural conditions. Only 336 seedlings, corresponding to 36 plant taxa, emerged under natural conditions, among which five taxa did not appear under greenhouse conditions. Graminoids and forbs composed 91% of the seedlings in the greenhouse whereas shrubs were more abundant under natural conditions, representing 55% of the emerged seedlings. Under greenhouse conditions, first red deer and then wild boar dispersed more species than the other two mammals, while under natural conditions brown bear was the most effective vector. We observed remarkably higher species richness and seedling abundance per dung sub-sample under buffered greenhouse conditions than we did under natural conditions. Conclusions: The four sympatric mammals studied provided different seed dispersal services, both in terms of seedling abundance and species richness and may therefore be regarded as complementary. Our results highlight a positive bias when only considering germination under buffered greenhouse conditions. This must be taken into account when planning management options to benefit plant biodiversity based on the dispersal services concluded from greenhouse experiments.


2012 ◽  
Vol 60 (4) ◽  
pp. 441-448 ◽  
Author(s):  
Diana Žele ◽  
Jana Avberšek ◽  
Igor Gruntar ◽  
Matjaž Ocepek ◽  
Gorazd Vengušt

Anaplasma phagocytophilumis a tick-borne rickettsial pathogen responsible for granulocytic anaplasmosis in mammalian hosts including humans. Wild animals may play an important role in the epidemiology of this disease. The aim of this study was to estimate the prevalence of infection withA. phagocytophilumamong wildlife in Slovenia. Serum samples (n = 376) from the most important game species [red deer (Cervus elaphus), roe deer (Capreolus capreolus), wild boar (Sus scrofa), chamois (Rupicapra rupicapra) and brown bear (Ursus arctos)] were examined byA. phagocytophilum-specific indirect fluorescent-antibody assay (IFA) and wild boar spleen samples (n = 160) were tested by polymerase chain reaction (PCR).A. phagocytophilum-specific antibodies were found in 72% of sera andA. phagocytophilumDNA was present in 6.2% of spleens. The data indicate thatA. phagocytophilumis present and widespread in Slovenian game animals and that game species are involved in the natural life cycle ofA. phagocytophilum.


2021 ◽  
Vol 14 (1) ◽  
Author(s):  
Nannet D. Fabri ◽  
Hein Sprong ◽  
Tim R. Hofmeester ◽  
Hans Heesterbeek ◽  
Björn F. Donnars ◽  
...  

Abstract Background Several ungulate species are feeding and propagation hosts for the tick Ixodes ricinus as well as hosts to a wide range of zoonotic pathogens. Here, we focus on Anaplasma phagocytophilum and Borrelia burgdorferi (s.l.), two important pathogens for which ungulates are amplifying and dilution hosts, respectively. Ungulate management is one of the main tools to mitigate human health risks associated with these tick-borne pathogens. Across Europe, different species of ungulates are expanding their ranges and increasing in numbers. It is currently unclear if and how the relative contribution to the life-cycle of I. ricinus and the transmission cycles of tick-borne pathogens differ among these species. In this study, we aimed to identify these relative contributions for five European ungulate species. Methods We quantified the tick load and collected ticks and spleen samples from hunted fallow deer (Dama dama, n = 131), moose (Alces alces, n = 15), red deer (Cervus elaphus, n = 61), roe deer (Capreolus capreolus, n = 30) and wild boar (Sus scrofa, n = 87) in south-central Sweden. We investigated the presence of tick-borne pathogens in ticks and spleen samples using real-time PCR. We determined if ungulate species differed in tick load (prevalence and intensity) and in infection prevalence in their tissue as well as in the ticks feeding on them. Results Wild boar hosted fewer adult female ticks than any of the deer species, indicating that deer are more important as propagation hosts. Among the deer species, moose had the lowest number of female ticks, while there was no difference among the other deer species. Given the low number of infected nymphs, the relative contribution of all ungulate species to the transmission of B. burgdorferi (s.l.) was low. Fallow deer, red deer and roe deer contributed more to the transmission of A. phagocytophilum than wild boar. Conclusions The ungulate species clearly differed in their role as a propagation host and in the transmission of B. burgdorferi and A. phagocytophilum. This study provides crucial information for ungulate management as a tool to mitigate zoonotic disease risk and argues for adapting management approaches to the local ungulate species composition and the pathogen(s) of concern. Graphic abstract


2019 ◽  
Author(s):  
Sorour Karimi ◽  
Mahmoud-Reza Hemami ◽  
Mostafa Tarkesh Esfahani ◽  
Christophe Baltzinge

Abstract Background: Among other factors, feeding regimes of different mammals affect their roles as endozoochorous seed dispersal vectors. We thus conducted a cross-species comparative approach involving two herbivores, red and roe deer; and two omnivores and opportunistic frugivores, wild boar and brown bear, which occupy the forest and prairie-forest ecotone habitats of the south-eastern Caspian region. To compare their potential and realised endozoochorous seed dispersal, we used two different seed germination methods: controlled greenhouse and natural conditions. Results: Of the 136 plant taxa (3078 seedlings) recorded from the 445 paired dung sub-samples of both methods, only 36 taxa (336 seedlings) were recorded under natural conditions, among which five taxa showed no emergence in the greenhouse. Graminoids and forbs composed 91% of the seedlings in greenhouse but shrub species were more abundant under natural conditions (55% of emerged seedlings). Considering greenhouse condition, red deer and then wild boar dispersed more species than the other animals, while in natural condition brown bear was the most effective vector. We observed remarkably higher species richness and seedling abundance in the controlled greenhouse method than natural condition (P≤0.001). This difference may be related to dung sample preparation method or meteorological conditions in the field during our experiment. Conclusions: The studied sympatric vectors were shown to provide different seed dispersal services and may, therefore, be regarded as complementary. Our results provide evidence for the potential positive bias arising when only considering germination under controlled greenhouse conditions. Relativising the potential versus the realised endozoochorous dispersal of seeds by different mammals would help determine management options to maintain plant biodiversity. Keywords : Different germination methods, Mammal functional groups, Potential endozoochory, Realised endozoochory


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