A labeling study of dentin formation rates during crown and root growth of porcine mandibular first molars.

We used fluorochrome labeling to study spatiotemporal variation of dentin apposition (DAR) and extension (DER) rates during crown and root formation of mandibular first molars from wild boar and domestic pigs. DAR was reconstructed along the course of dentinal tubules in four zones of the crown and in the upper root area. In all five zones, mean DAR increased during the first 30% to 40% of apposition, reaching highest values (22-23 µm/day) in the upper-lateral crown zone. Lowest values were recorded near the dentin-pulp interface (DPI). Typically, DARs in contemporaneously formed dentin areas were higher in more cuspally compared to more cervically/apically located zones. DER was high (>200 µm/day) in early postnatal crown dentin and then decreased markedly in cervical direction, with lowest values in the cervical crown zone. After this nadir, DER sharply increased in the upper 30% to 40% of the root extension, reaching values equaling (wild boar) or even surpassing (domestic pigs) those recorded in the upper lateral crown. After this peak, DER again decreased. While DAR did not differ markedly between wild boar and domestic pigs, the DER showed marked differences, both regarding maximum values (208.1 µm/day in wild boar, 272.2 µm/day in domestic pigs) and the timing of the root growth spurt, which occurred earlier in the domestic pigs. We consider the more rapid recruitment of secretory odontoblasts in domestic pigs (reflected by higher DER) a side effect of selection for rapid body growth during pig domestication.

Dose-dependent in vivo effects of formulated moxidectin on seedling emergence of temperate grassland species.

Sheep function as effective endozoochorous seed vectors in grasslands. Recent laboratory-based studies showed that this important function can be impaired by macrocyclic lactone anthelmintics, which are used to control parasites and enter into the environment mainly via faeces; however, there is a lack of in vivo studies. We conducted a seed-feeding experiment with sheep that included four temperate grassland species from four different families (Achillea ptarmica, Asteraceae; Agrostis capillaris, Poaceae; Dianthus deltoides, Caryophyllaceae; Plantago lanceolata, Plantaginaceae). A series of three feeding trials was carried out after one of two groups of sheep received a single administration of a common oral formulation of the macrocyclic lactone moxidectin. Faeces were collected to determine seedling emergence rate and emergence timing as well as moxidectin concentration via HPLC. Seedling emergence differed significantly between the anthelmintic-treated sheep and the control group. This impact depended on time of seed uptake after anthelmintic administration. Number of emerging seedlings was significantly reduced (27.1 %) when faeces moxidectin concentrations were high (on average 3153 ng g-1; 1 d post treatment) and significantly increased (up to 68.8 %) when moxidectin concentrations were low (≤86 ng g-1; 7, 14 d pt). Mean emergence time was significantly lowered at low moxidectin concentrations. These results demonstrate dose-related effects of deworming on seedling emergence which might affect endozoochory and eventually plant population dynamics in grasslands.

First Insights into the Occurrence of Circular Single-Stranded DNA Genomes in Asian and African Cattle.

Circular replicase-encoding single-stranded (CRESS) DNA viruses and other circular DNA agents are increasingly found in various samples and animals. A specific class of these agents-termed bovine meat and milk factors (BMMF)-has been supposed to act as a factor in indirect carcinogenesis in humans. Initial observations attributed the BMMF to European cattle breeds and foodstuffs produced thereof. In the present study, blood and fecal samples from African and Asian cattle were examined. BMMF molecules and genomoviruses were detected in all bovids under study. The majority (79%) of the 29 circular elements could be assigned to BMMF groups 1 and 2, whereas CRESS viruses of the family Genomoviridae accounted for the smaller part (21%). Two genomoviruses belong to the genus Gemykibivirus and one to the genus Gemykrogvirus. The remaining three might be considered as novel species within the genus Gemycircularvirus. The majority of all isolated molecules originated from fecal samples, whereas only three derived from blood. The results from this study expand our knowledge on the diversity and presence of circular DNA in different ruminants that serve for food production in many countries over the world.

Wild boar versus domestic pig-Deciphering of crown growth in porcine second molars.

Based on the previously established periodicity of enamel growth marks, we reconstructed crown growth parameters of mandibular second molars from two wild boar and two domestic pigs of the Linderöd breed. Body weight gain and progression of dental development were markedly faster in the domestic pigs than the wild boar. While the final crown dimensions of the M2 did not differ between domestic pigs and wild boar, mean crown formation time (CFT) of this tooth was considerably shorter in the domestic pigs (162 days) than in the wild boar (205 days). The difference in CFT was mainly attributable to a higher enamel extension rate (EER) in the domestic pig. Generally, EER was highest in the cuspalmost deciles of the length of the enamel-dentine-junction and markedly dropped in cervical direction, with lowest values occurring in the cervicalmost decile. In consequence, the cuspal half of the M2 crown was formed about three times faster than the cervical half. In contrast to the EER, no marked difference in daily enamel secretion rate (DSR) was recorded between domestic pigs and wild boar. The duration of enamel matrix apposition as well as linear enamel thickness in corresponding crown portions was only slightly lower in the domestic pigs than the wild boar. Thus, the earlier completion of M2 crown growth in the domestic pig was mainly achieved by a higher EER and not by an increased DSR. The more rapid recruitment of secretory ameloblasts in the course of molar crown formation of domestic pigs compared to wild boar is considered a side-effect of the selection for rapid body growth during pig domestication.

Ecology impacts the decrease of Spirochaetes and Prevotella in the fecal gut microbiota of urban humans.

Compared to the huge microbial diversity in most mammals, human gut microbiomes have lost diversity while becoming specialized for animal-based diets - especially compared to chimps, their genetically closest ancestors. The lowered microbial diversity within the gut of westernized populations has also been associated with different kinds of chronic inflammatory diseases in humans. To further deepen our knowledge on phylogenetic and ecologic impacts on human health and fitness, we established the herein presented biobank as well as its comprehensive microbiota analysis. In total, 368 stool samples from 38 different animal species, including Homo sapiens, belonging to four diverse mammalian orders were collected at seven different locations and analyzed by 16S rRNA gene amplicon sequencing. Comprehensive data analysis was performed to (i) determine the overall impact of host phylogeny vs. diet, location, and ecology and to (ii) examine the general pattern of fecal bacterial diversity across captive mammals and humans.By using a controlled study design with captive mammals we could verify that host phylogeny is the most dominant driver of mammalian gut microbiota composition. However, the effect of ecology appears to be able to overcome host phylogeny and should therefore be studied in more detail in future studies. Most importantly, our study could observe a remarkable decrease of Spirochaetes and Prevotella in westernized humans and platyrrhines, which is probably not only due to diet, but also to the social behavior and structure in these communities.Our study highlights the importance of phylogenetic relationship and ecology within the evolution of mammalian fecal microbiota composition. Particularly, the observed decrease of Spirochaetes and Prevotella in westernized communities might be associated to lifestyle dependent rapid evolutionary changes, potentially involved in the establishment of dysbiotic microbiomes, which promote the etiology of chronic diseases.

Characterization of short-period and long-period incremental markings in porcine enamel and dentine-Results of a fluorochrome labelling study in wild boar and domestic pigs.

Mammalian dental hard tissues exhibit incremental markings that reflect the periodic variation of appositional growth rates. In order to use these markings to characterize dental growth processes and to infer life-history traits, an unequivocal identification of their periodicities is required. We performed a fluorochrome labelling study on forming enamel and dentine in molar teeth of wild boar and domestic pigs to establish the periodicity and temporal correspondence of incremental markings in enamel and dentine. The dominant incremental markings in enamel (laminations) and dentine (von Ebner lines) recorded in the pig teeth are of a daily nature. In addition, long-period incremental markings with a periodicity of 2 days were recorded in enamel (striae of Retzius) and dentine (Andresen lines). The 2-day growth rhythm was also expressed at the lateral crown surface, as evidenced by the pattern of perikymata. In enamel, also markings with a sub-daily periodicity, representing an ultradian growth rhythm, were observed. Our study provides experimental evidence for the periodicity of incremental markings in porcine enamel and dentine. The findings correct previous misconceptions on incremental markings in dental hard tissues of pigs and other ungulates that had led to erroneous conclusions regarding crown formation parameters.

Ontogenetic changes of tissue compartmentalization and bone type distribution in the humerus of Soay sheep.

We studied ontogenetic changes of histomorphological features and bone type distribution in the humeral midshaft region of Soay sheep from three postnatal age classes (13, 25, and 33 months). Our study demonstrated a marked change of bone type distribution in the humeri with age. In the cortical midshaft region of 13-month-old individuals, periosteal fibrolamellar bone was the dominating bone type. This indicates a rapid bone growth during the first year of life, which was only interrupted by a seasonal growth arrest in the animals' first winter. In individuals from the two older age classes, periosteal lamellar-zonal bone and intermediate fibrolamellar bone had been formed at the periosteal surface, and endosteal lamellar-zonal bone at the endosteal surface. These bone types are indicative of a reduced bone growth rate. A marked reduction in radial growth was already recorded in the 25-month-old individuals. Distribution and extent of secondary bone showed a marked bilateral symmetry in the humeri of individual sheep. The presence of secondary bone was largely restricted to the anterior (cranial) and the medial cortical areas. This characteristic distribution of remodeling activity within the humeral cortex of sheep is consistent with the view that remodeling activity is largely caused by compressive stress. Our study further demonstrated the presence of a considerable cortical drift in the sheep humeri over the study period, with endosteal resorption occurring predominantly in the posterior (caudal) quadrant and formation of a prominent endosteal lamellar pocket in the anterior (cranial) and medial cortical quadrants.

The pay-off of hypsodonty - timing and dynamics of crown growth and wear in molars of Soay sheep.

BACKGROUND: Several lineages of herbivorous mammals have evolved hypsodont cheek teeth to increase the functional lifespan of their dentition. While the selective drivers of this trend and the developmental processes involved have been studied in greater detail, thus far no quantitative information is available on the relationship between additional investment into tooth growth and the resulting extension of the functional period of these teeth. To achieve this, we performed a detailed analysis of molar crown growth in known-age Soay sheep repeatedly injected with different fluorochromes. RESULTS: Our study revealed that in sheep molars especially the formation of the crown base portion is prolonged in comparison with other herbivorous artiodactyl species. Our results demonstrate that growth of the crown base accounted for more than half of the total crown formation time (CFT) of the anterior lobes of the first (approx. 220 days of total CFT of 300 days), second (approx. 260 of 460 days) and third (approx. 300 of at least 520 days) molars, and that the formation of this crown portion occurred largely after the teeth had already reached functional occlusion. By combining data on wear-related changes in crown morphology from the literature with the reconstructed additional investment into the crown base portion, it was possible to relate this additional investment to a prolongation of the functional periods of the molars ranging from 4 years in the M1 to 6 years in the M3. CONCLUSIONS: Our results allow to establish a quantitative link between an additional investment into molar crown growth of sheep and the extension of the functional period of these teeth. The reported findings enable an assessment of the adaptive value, in terms of increased longevity, of an additional investment into crown elongation in a mammalian herbivore.

Mineral Apposition Rates in Coronal Dentine of Mandibular First Molars in Soay Sheep: Results of a Fluorochrome Labeling Study.

We studied the spatio-temporal variation of mineral apposition rate (MAR) in postnatally formed coronal dentine of mandibular first molars from Soay sheep repeatedly injected with different fluorochromes. MAR declined along the cuspal to cervical crown axis, and from early to late formed dentine, that is, from the dentine at the enamel-dentine-junction (EDJ) to the dentine adjacent to the dentine-pulp-interface (DPI). Highest mean MARs (about 21 µm/day) were recorded in cuspal dentine formed in the period of 28-42 days after birth. Lowest values (<2 µm/day) were recorded in late-formed (secondary) dentine close to the DPI. The high MARs recorded in the dentine of the cuspal crown portions enable the formation of a large tooth crown within a relatively short period of less than one year. The established MARs in the dentine of the different crown portions of sheep molars will allow a precise determination of the timing of stress events affecting dentine formation. They are also helpful for devising sampling protocols in studies of trace element or stable isotope distributions in sheep dentine aimed at assessing temporal variation of incorporation into forming dentine. Such data are useful in a variety of contexts, including, for example, the exposure to pollutants and the reconstruction of husbandry practices or feeding regimes. Anat Rec, 301:902-912, 2018. © 2017 Wiley Periodicals, Inc.

Endogenous Retroviral Insertions Indicate a Secondary Introduction of Domestic Sheep Lineages to the Caucasus and Central Asia between the Bronze and Iron Age.

Sheep were one of the first livestock species domesticated by humans. After initial domestication in the Middle East they were spread across Eurasia. The modern distribution of endogenous Jaagsiekte sheep retrovirus insertions in domestic sheep breeds suggests that over the course of millennia, successive introductions of improved lineages and selection for wool quality occurred in the Mediterranean region and most of Asia. Here we present a novel ancient DNA approach using data of endogenous retroviral insertions in Bronze and Iron Age domestic sheep from the Caucasus and Pamir mountain areas. Our findings support a secondary introduction of wool sheep from the Middle East between the Late Bronze Age and Iron Age into most areas of Eurasia.

[The importance of conservation of rare domestic animal breeds using the example of Arc Warder]. / Die Bedeutung der Erhaltung vom Aussterben bedrohter Nutztierrassen am Beispiel der Arche Warder.

The domestic animals descend from various wild ancestors. Thus, for example, of the wolf, the dog (15  000 BC), of the Bezoar goat the goat (10  000 BC), of the Asian mouflon the sheep (10  000 BC), of the wild boar the pig (8000 BC) and of the aurochs the cattle (7000 BC). Domestication has dramatically changed our culture and led to a great diversity of animal breeds. This is a unique cultural and historical treasure, which we have to preserve for future generations. The zoological park Arc Warder is Europe's largest center for rare and endangered domestic animal breeds. Arc Warder is more than a zoo; it is a landscape park, a conservation venture for genetic resources and furthermore a research project. Five principles guide the conservation efforts of Arc Warder: 1. Conservation through preservation. The breeding program will improve the quality of the genetic reservoir of breeds. 2. Conservation through the establishment of satellite stations. These pastures outside the park allow to increase the gene pool and ensure the protection of breeds against epidemics. 3. Conservation through high level education. 4. Protection by networking with national and international institutions. 5. Protection through research. Arc Warder is actively involved in close scientific cooperation with various universities and other research institutions on a number of scientific projects concerning the biological characteristics of old breeds.

Lines of evidence-incremental markings in molar enamel of Soay sheep as revealed by a fluorochrome labeling and backscattered electron imaging study.

We studied the structural characteristics and periodicities of regular incremental markings in sheep enamel using fluorochrome injections for vital labeling of forming enamel and backscattered electron imaging in the scanning electron microscope. Microscopic analysis of mandibular first molars revealed the presence of incremental markings with a daily periodicity (laminations) that indicated successive positions of the forming front of interprismatic enamel. In addition to the laminations, incremental markings with a sub-daily periodicity were discernible both in interprismatic enamel and in enamel prisms. Five sub-daily increments were present between two consecutive laminations. Backscattered electron imaging revealed that each sub-daily growth increment consisted of a broader and more highly mineralized band and a narrower and less mineralized band (line). The sub-daily markings in the prisms of sheep enamel morphologically resembled the (daily) prisms cross striations seen in primate enamel. Incremental markings with a supra-daily periodicity were not observed in sheep enamel. Based on the periodicity of the incremental markings, maximum mean daily apposition rates of 17.0 µm in buccal enamel and of 13.4 µm in lingual enamel were recorded. Enamel extension rates were also high, with maximum means of 180 µm/day and 217 µm/day in upper crown areas of buccal and lingual enamel, respectively. Values in more cervical crown portions were markedly lower. Our results are in accordance with previous findings in other ungulate species. Using the incremental markings present in primate enamel as a reference could result in a misinterpretation of the incremental markings in ungulate enamel. Thus, the sub-daily growth increments in the prisms of ungulate enamel might be mistaken as prism cross striations with a daily periodicity, and the laminations misidentified as striae of Retzius with a supra-daily periodicity. This would lead to a considerable overestimation of crown formation times in ungulate teeth.

Mycobacterial infections in free-living cervids in Germany (2002-2006).

We examined 1,022 free-living roe deer, red deer, and fallow deer for mycobacteria in Germany, 2002-2006. Retropharyngeal lymph nodes and other tissues were processed for culture and isolates were identified with the use of polymerase chain reaction and DNA sequencing. Mycobacteria were found in 18.3% of deer, with Mycobacterium avium in 14.8%. Other atypical mycobacteria were detected in 5.3%. Members of the Mycobacterium tuberculosis complex were not detected.

Experimental superinfection of a Lesser Malayan mousedeer (Tragulus javanicus) persistently infected with bovine viral diarrhea virus.

A Lesser Malayan mousedeer (Tragulus javanicus), persistently infected with noncytopathogenic bovine viral diarrhea virus (BVDV) type 1f, was experimentally superinfected with a cytopathogenic isolate of BVDV type 1c, which antigenically partially matched the endogenous strain. Within the observational period of 125 days after superinfection, the animal did not demonstrate any clinical signs of the disease and/or significant changes in blood values. Neutralizing antibodies were detected at 35 and 42 days postinfection. The isolate causing the superinfection was found in feces, nasal swabs, and saliva starting from day 29 and at various times postchallenge. Macroscopic or histologic examination did not reveal mucosal disease-like lesions, despite the detection of the cytopathogenic isolate in the salivary gland, rumen, abomasum, kidney, and superficial prescapular lymph node. Results indicate that the cytopathogenic BVDV strain, which was used in the superinfection, persisted in the viremic animal without causing disease within the observation period.

Pinching off syndrome in free-ranging white-tailed sea eagles (Haliaeetus albicilla) in Europe: frequency and geographic distribution of a generalized feather abnormality.

Generalized feather abnormalities are rarely documented in free-living birds. Pinching off syndrome (POS) is a feather abnormality in which all remiges and retrices become malformed and are lost during the nestling stage, rendering the bird unable to fly. To determine the frequency of occurrence and geographic distribution of this syndrome in white-tailed sea eagles (Haliaeetus albicilla) in Europe, we sent questionnaires to ornithologists in 19 countries within the European range of this species. We also searched for reported cases of sea eagles with feather abnormalities that met the criteria of POS. Overall, 32 nestlings or fledglings with POS were identified between 1975 and 2006. The geographic distribution of cases was primarily restricted to 4 European countries: Germany (17 cases), Poland (11 cases), the Czech Republic (3 cases), and Great Britain (1 case). Eleven eagles from Germany and 2 eagles from the Czech Republic were examined clinically. In 15 birds in which sex was determined, 8 were female and 7 were male. From 2000 to 2005, the 5-year incidence of POS in white-tailed sea eagles in Germany was 3.5 cases per 1000 birds. Although the etiology of this syndrome in wild sea eagles is unknown, our results support a possible genetic cause.

Investigations on the aetiology of pinching off syndrome in four white-tailed sea eagles (Haliaeetus albicilla) from Germany.

The purpose of this study was to investigate the aetiology of the pinching off syndrome (POS), a generalized feather abnormality affecting free-living nestling of the white-tailed sea eagle (Haliaeetus albicilla) in Europe. For the first time, extensive clinical, haematological, biochemical, virological, bacteriological, nutritional, histopathological, parasitological and electron microscopical examinations were performed on three females and one male suffering from POS. Early and increased cytokeratin formation at the base of regenerating feathers and their follicle was observed in affected birds. Ultrathin sections of the feather papillae revealed an extended stratum transitivum and a compact, thickened keratinized stratum corneum. The transitional cells in POS feathers contained vacuoles often associated with the nucleus. Lipofuscin accumulations in neurons, glial cells and islet cells of the pancreas were found in all examined birds. It was not clear whether there is an association between the occurrence of lipofuscin and POS. No evidence was found to suggest that infectious agents (parasites, bacteria, fungi or viruses), malnutrition or hormonal imbalances are involved in the aetiology of POS in white-tailed sea eagles. It remains unclear whether there is a genetic background of POS.

New variants of European brown hare syndrome virus strains in free-ranging European brown hares (Lepus europaeus) from Slovakia.

Investigations regarding European brown hare syndrome virus (EBHSV) in European brown hares (Lepus europaeus) in Slovakia were undertaken in order to detect the possible presence of EBHSV and to evaluate its phylogenetic position. Liver and/or serum samples were obtained from 135 European brown hares shot by hunters in eight regional hunting areas. From 36 animals corresponding liver and serum samples were available; from the remaining 49 and 50 animals only liver or serum samples were examined, respectively. Samples were tested for antibodies against EBHSV and for viral RNA by reverse transcriptase-polymerase chain reaction (RT-PCR) and RT-PCR products were subsequently sequenced. Additionally, matrilinear hare haplotypes were analyzed in order to detect potential familial susceptibility to EBHSV. Sixty-three of 86 sera contained antibodies against EBHSV, whereas 15 of 85 liver samples were PCR positive. Of the latter, 14 were sequenced, revealing three new strains of EBHSV. Fifteen different matrilinear haplotypes were identified, but no correlation was found between haplotype and susceptibility to EBHSV infection. Our findings confirmed the existence of EBHSV in Slovakia and reinforce the need for determining EBHSV status when importing hares for restocking.

Progressive ataxia in a captive North American river otter (Lontra canadensis) associated with brain stem spheroid formation.

Progressive ataxia, particularly of the hind limbs, developed in a paretic female juvenile captive North American river otter (Lontra canadensis). Severe bilaterally symmetrical axonal degeneration in the medulla oblongata (nucleus gracilis and nucleus cuneatus) and multiple scattered spheroids in the region of the pons were detected microscopically. This is the first description of ataxia associated with axonal degeneration in otters.

Zoo and wildlife medical education: a European perspective.

Europe has a long and distinguished history in veterinary science and education, and it was here that the first professional investigations of pathological conditions in zoo animals took place. However, despite an increasing number of veterinarians working with wildlife, education in zoological and wildlife medicine has only recently become part of formal veterinary training at the undergraduate level. Consequently, current educational opportunities in zoological and wildlife medicine vary widely throughout Europe, both in availability and in composition. The need to establish agreed standards in education across Europe and to foster the mobility of students and teaching staff are reflected by international agreements such as the Bologna Treaty and the ERASMUS-SOCRATES program. Europe is also home to a number of voluntary professional organizations, such as the European Wildlife Disease Association and the European Association of Zoo and Wildlife Veterinarians, that actively promote the inclusion of zoological and wildlife medicine in veterinary education. Zoo and wildlife medicine is currently a veterinary specialization in Europe, but educational opportunities are likely to increase in availability in the future.

Characterization of equid herpesvirus 1 (EHV-1) related viruses from captive Grevy's zebra and blackbuck.

Equid herpes virus 1 (EHV-1) related isolates from a captive blackbuck (strain Ro-1) and Grevy's zebra (strain T965) behaved similarly to EHV-1 and EHV-9 in respect to their host cell range. Restriction enzyme analysis and a phylogenetic tree confirmed that Ro-1 and T965 were identical and more closely related to EHV-1 than to EHV-9. Differences from EHV-1 became obvious firstly, by amino acid alignments revealing two unique substitutions in the gB protein of Ro-1 and T965. Secondly, an EHV-1 type-specific monoclonal antibody did not detect its antigen on Ro-1, T965 or EHV-9 infected cells by immunohistochemistry. The results support the view that Ro-1 and T965 isolates represent a distinct, previously unrecognized species of equid herpesviruses.