Ecology of uncultivated Oscillospira species in the rumen of cattle, sheep, and reindeer as assessed by microscopy and molecular approaches.
ABSTRACT: The ecology of the uncultured, but large and morphologically conspicuous, rumen bacterium Oscillospira spp. was studied. Oscillospira-specific 16S rRNA gene sequences were detected in North American domestic cattle, sheep from Australia and Japan, and Norwegian reindeer. Phylogenetic analysis of the sequences obtained allowed definition of three operational taxonomic units within the Oscillospira clade. Consistent with this genetic diversity, we observed atypical smaller morphotypes by using an Oscillospira-specific fluorescence in situ hybridization probe. Despite the visual disappearance of typical large Oscillospira morphotypes, the presence of Oscillospira spp. was still detected by Oscillospira-specific PCR in the rumen of cattle and sheep. These observations suggest the broad presence of Oscillospira species in various rumen ecosystems with the level, and most likely the morphological form, dependent on diet. An ecological analysis based on enumeration of the morphologically conspicuous, large-septate form confirms that the highest counts are associated with the feeding of fresh forage diets to cattle and sheep and in two different subspecies of reindeer investigated.
Project description:BACKGROUND: Two outbreaks of the disease contagious ecthyma were reported in 1999 and 2000 in Norwegian semi-domesticated reindeer (Rangifer tarandus tarandus). Contagious ecthyma is an epidermal disease of sheep and goats worldwide, which is caused by the zoonotic parapoxvirus orf virus. Characterisation of clinical samples from the two outbreaks in semi-domesticated reindeer in Norway by electron microscopy and PCR (B2L) revealed typical parapoxvirus particles and partial gene sequences corresponding to parapoxvirus, respectively. If contagious ecthyma in reindeer is caused by orf virus, the virus may be transferred from sheep and goats, via people, equipment and common use of pastures and corrals, to reindeer. Another possibility is that contagious ecthyma in reindeer is caused by a hitherto unclassified member of the parapoxvirus genus that circulates among reindeer herds and remains endemic in Norway. RESULTS: Genomic comparisons of one standard orf strain (orf NZ2) and the reindeer isolates, employing restriction fragment length polymorphism (RFLP) and random amplified polymorphic DNA (RAPD) analysis, demonstrated high similarity between the reindeer viruses and known orf virus strains. Partial DNA sequences of two different viral genes were determined for the different isolates and compared with corresponding parapoxvirus genebank sequences. The comparison/alignment and construction of phylogenetic trees also point to an affiliation of the reindeer viruses to the species orf virus. CONCLUSION: The results of this work imply that the parapoxvirus causing contagious ecthyma in Norwegian semi-domesticated reindeer belongs to the species orf virus and that the orf virus crosses the host species barrier from sheep and goat to semi-domesticated reindeer.
Project description:Reindeer (Rangifer tarandus tarandus) are large Holarctic herbivores whose heterogeneous diet has led to the development of a unique gastrointestinal microbiota, essential for the digestion of arctic flora, which may include a large proportion of lichens during winter. Lichens are rich in plant secondary metabolites, which may affect members of the gut microbial consortium, such as the methane-producing methanogenic archaea. Little is known about the effect of lichen consumption on the rumen and cecum microbiotas and how this may affect methanogenesis in reindeer. Here, we examined the effects of dietary lichens on the reindeer gut microbiota, especially methanogens. Samples from the rumen and cecum were collected from two groups of reindeer, fed either lichens (Ld: n = 4), or a standard pelleted feed (Pd: n = 3). Microbial densities (methanogens, bacteria and protozoa) were quantified using quantitative real-time PCR and methanogen and bacterial diversities were determined by 454 pyrosequencing of the 16S rRNA genes. In general, the density of methanogens were not significantly affected (p>0.05) by the intake of lichens. Methanobrevibacter constituted the main archaeal genus (>95% of reads), with Mbr. thaueri CW as the dominant species in both groups of reindeer. Bacteria belonging to the uncharacterized Ruminococcaceae and the genus Prevotella were the dominant phylotypes in the rumen and cecum, in both diets (ranging between 16-38% total sequences). Bacteria belonging to the genus Ruminococcus (3.5% to 0.6%; p = 0.001) and uncharacterized phylotypes within the order Bacteroidales (8.4% to 1.3%; p = 0.027), were significantly decreased in the rumen of lichen-fed reindeer, but not in the cecum (p = 0.2 and p = 0.087, respectively). UniFrac-based analyses showed archaeal and bacterial libraries were significantly different between diets, in both the cecum and the rumen (vegan::Adonis: pseudo-F<0.05). Based upon previous literature, we suggest that the altered methanogen and bacterial profiles may account for expected lower methane emissions from lichen-fed reindeer.
Project description:Studies of locomotor activity in Svalbard reindeer reported the temporary absence of diel rhythms under Arctic photic conditions. However, using Lomb-Scargle periodogram analyses with high statistical power we found diel or circadian rhythmicity throughout the entire year in measures of behaviour, temperature in the rumen and heart rate in free-living Svalbard reindeer. Significant diel rhythmicity was only lacking during some of the 15-day intervals analysed in the less frequently measured heart rate. During Polar Night these rhythms were free-running and attenuated. During continual daylight in summer, rhythms where entrained to 24?hours corresponding with the daily variation in the intensity of solar radiation, but attenuated when continuous daylight coincided with the period of growing forage. Diel rhythmicity was reduced during this short period of peak foraging activity, which coincided with peak heart rate and temperature in the rumen, most likely to facilitate fattening when food is abundant. For the rest of the year, heart rate and temperature showed the most pronounced and long-lasting suppression ever found in ungulates. The profound seasonal changes in foraging, metabolic activity, and power of diel and circadian rhythmicity of Svalbard reindeer can be viewed as adaptations to the extreme living conditions in the High Arctic.
Project description:Members of the viral subfamily Alphaherpesvirinae establish latency from which they can be reactivated. Bovine herpesvirus 1 causes infectious bovine rhinotracheitis and infectious pustular vulvovaginitis in cattle, as well as abortion and weak calves. Serological evidence of alphaherpesvirus infection has been reported for wild and semidomesticated reindeer (Rangifer tarandus tarandus) in Norway. To address the possibility that reindeer alphaherpesvirus (cervid herpesvirus 2 [CvHV-2]) infection might affect the respiratory system and in part explain the relatively high mortality of reindeer calves during their first year, tissue samples were obtained from reindeer and reindeer fetuses at slaughterhouses in Finnmark County, Norway. A nested pan-alphaherpesvirus PCR amplification targeting the highly conserved UL27 gene (encoding glycoprotein B) was used. Sequencing of amplicons revealed the presence of CvHV-2 DNA. The detection of CvHV-2 DNA in trigeminal ganglia (27 of 143 samples), nasal swabs (5 of 75 samples), and fetal tissues (12 of 48 samples) indicates that CvHV-2 infection is endemic in this reindeer population. Moreover, the virus is transmitted horizontally by the respiratory route, establishing latency in the trigeminal ganglion, and vertically to the fetus through the placenta. Further studies should focus on the reproductive impact of CvHV-2 infection in reindeer.
Project description:Nematodes of the genera Elaphostrongylus and Dictyocaulus are associated with disease in semi-domesticated tundra reindeer and farmed red deer whereas less knowledge exists in the wild. Their first stage larvae (L1) develop to the infective third stage (L3) in the environment; Elaphostrongylus spp. within intermediate gastropod hosts and Dictyocaulus spp. as free-living larvae. Larval development of Elaphostrongylus is highly temperature dependent with a developmental minimum of 9-10?°C. Larval development of Dictyocaulus spp. may occur at low temperatures (5?°C) but the larvae are sensitive to desiccation. We examined the prevalence and intensity of Elaphostrongylus spp. and Dictyocaulus spp. infections in six wild reindeer and two wild red deer populations in relation to altitude, temperature and rainfall in their respective main summer pasture area over the 5 summers prior to sampling. The parasitological examination was based upon morphological identification of L1 in the faeces of hunted animals. Altitude was calculated from animal position data and temperature and precipitation by means of a nationwide gridded data set. Temperature decreased with increasing altitude, from 13.3?°C for the lowest located red deer population (300?m) to 6.1?°C for the highest located reindeer population (1400?m). No significant relationship between altitude and rainfall was identified. Elaphostrongylus spp. infection decreased in prevalence with increasing altitude, being identified in 89% of investigated samples from the lowest located population and in 3% of samples from the highest. The prevalence of Dictyocaulus spp. infection varied between 28 and 80% and no relationship with altitude was found. The intensity of Elaphostrongylus spp. infection was low in reindeer and moderate in red deer whereas the intensity of Dictyocaulus spp. infection was moderate in both species. Our results indicated that the climatic conditions in all areas studied were suitable for Dictyocaulus spp., whereas summer temperature was a restrictive factor for Elaphostrongylus sp. in reindeer.
Project description:We evaluated the genetic diversity of samples identified morphologically as Fasciola spp. from sheep, cattle and goat from Kermanshah Province, western Iran using PCR-RFLP method.We used PCR-RFLP analysis of ribosomal ITS1 fragment using RsaI restriction enzyme to investigate the genetic characteristics of Fasciola species obtained from different hosts (16 sheep, 28 cattle, 4 goats). The species of Fasciola were confirmed by sequencing the 700 bp region of ribosomal ITS1 gene.In Kermanshah, F. hepatica was present in 96% of the samples, F. gigantica was found only in two cattle sample. No hybrid forms were detected in the present study.Our results contribute to clarify the dark spots of Fasciola genotyping in different parts of Iran.
Project description:Captive reindeer in German zoos and wildlife parks live outside their natural geographic range and are exposed to a variety of viral, bacterial and protozoan pathogens, some host-specific and some which they are not exposed to in their native habitat. Reindeer blood samples and ticks collected in 2013 from 123 reindeer at 16 different zoological facilities were available from a previous study. The aims of this study were to assess the serological status of these animals with regards to various microorganisms as well as to test ticks (Ixodes ricinus) and blood samples for the presence of Anaplasma spp. DNA in order to evaluate the exposure of captive reindeer in Germany to a variety of pathogens. A total of 119 or 118 serum samples were screened (ELISA) and antibodies were detected (seropositive/tested, prevalence, confidence interval) against alphaherpesvirus (24/119, 20.3%, CI: 13.9-28.3), bluetongue virus (BTV; 4/119, 3.4%, CI: 1.0-8.7), malignant catarrhal fever related gammaherpesvirus (MCFV-related gammaherpesvirus; 7/119, 5.9%, CI: 2.7-11.9), pestivirus (5/118, 4.2%, CI: 1.6-9.8), Schmallenberg virus (SBV; 70/118, 59.3%, CI: 50.3-67.8), smooth Brucella spp. (1/118; 0.9%, CI: 0-5.1), Neospora caninum (5/118, 4.2%, CI: 1.6-9.8), and Toxoplasma gondii (62/119, 52.1%, CI: 43.2-60.9). These results suggested the exposure of reindeer to all tested pathogens. Moreover, real-time PCR for Anaplasma phagocytophilum targeting the partial msp2 gene was performed on DNA extracted from whole blood samples from reindeer (n = 123) and from ticks (n = 49) collected from 22 reindeer in seven different facilities. In addition to the real-time PCR, a semi-nested PCR for the partial groEL gene, and a nested PCR targeting the partial 16S rRNA gene were performed. DNA of A. phagocytophilum was detected in 17 reindeer (13.8%) and 15 ticks (30.6%). Three of the five reindeer with ticks having A. phagocytophilum DNA also had such DNA in blood. These results indicate that captive reindeer can be exposed to several ruminant pathogens that they hitherto had no known exposure to through their natural geographical distribution and habitats as shown for Culicoides-borne BTV and SBV. Further, captive reindeer may serve as reservoir hosts for pathogens circulating in local domestic, captive, and wild ruminant species and populations and arthropod vectors.
Project description:Supplementary winter feeding of semi-domesticated reindeer (Rangifer tarandus tarandus) has become more common in Sweden and Norway due to reindeer pasture fragmentation and climatic conditions. With increased corralling and feeding, often associated with animal stress, increased animal-to-animal contact, and poor hygienic conditions, an altered range of health challenges and diseases may emerge. An outbreak of three different infectious diseases appeared simultaneously in a reindeer herd in Norrbotten County, Sweden. The animals were corralled and fed silage. Several animals in poor body condition stopped eating, with drool and discoloration of the hair coat around the mouth. There were large, black, necrotic lesions on the tongue and gingiva, with holes perforating the chin, indicative of oral necrobacillosis and Fusobacterium spp. infection. Simultaneously, animals were seen with proliferative lesions in the oral mucosa and on the lips, characteristic of contagious ecthyma and Orf virus infection. Furthermore, three animals had keratoconjunctivitis suggesting exposure to cervid herpesvirus 2 (CvHV2) and possibly secondary bacterial infections. DNA specific for Fusobacterium necrophorum and ORFV was detected in relevant tissue samples. Antibodies against CvHV2 were detected in 10 of 13 diseased and in four of 11 apparently healthy reindeer. Nine animals were found dead or were euthanized during the outbreak. Health risk factors associated with feeding and corralling may severely impact animal welfare and the herder's economy, and may represent an underestimated cost when replacing natural grazing with feeding.
Project description:This study demonstrated the potential effects of the rumen microbiota on the deposition of intramuscular fat, known as marbling. Previous studies on fatty acid metabolism in beef cattle have mostly focused on biohydrogenating rumen bacteria, whereas those on the overall rumen microbiota-to understand their roles in marbling-have not been systematically performed. The rumen microbiota of 14 Korean beef cattle (Hanwoo), which showed similar carcass characteristics and blood metabolites but different marbling scores, were analyzed by 16S rRNA gene sequencing. The rumen samples were grouped into two extreme marbling score groups of host animals as follows: LMS, marbling score? 4 or HMS, marbling score ?7. Species richness tended to be higher in the HMS group, whereas the overall microbiota differed between LMS and HMS groups. RFP12, Verrucomicrobia, Oscillospira, Porphyromonadaceae, and Paludibacter were differentially abundant in the HMS group, whereas Olsenella was abundant in the LMS group. Some marbling-associated bacterial taxa also contributed to the enrichment of two lipid metabolic pathways including "alpha-linolenic acid metabolism" and "fatty acid biosynthesis" in the HMS microbiome. Taxonomic drivers of fatty acid biosynthesis, particularly in the rumen microbiome of high-marbled meat, could thus be further studied to increase the intramuscular fat content.
Project description:Reindeer species (Rangifer tarandus, Linnaeus, 1758) includes wild and semi-domesticated ruminants belonging to Capreaolinae subfamily of Cervidae family reared in Eurasia (reindeer subspecies) and North America (caribou subspecies). Herding of reindeer has a great historical, socio-economic and ecological importance, especially to indigenous ethnic minorities. Infectious disease threats may therefore impact not solely the animal population driving it to further extinction and irreversible alterations to the wild environments of northern hemisphere, but also add to cultural changes observed as negative impact of globalization. Introduction of new technologies to control of reindeer migration between dwindling pasture areas and intensification of reindeer husbandry may facilitate the intra- and interspecies transmission of pathogens. The role of the reindeer as a potential BVDV reservoir has been studied, however, the number of publications is rather limited. The observed seroprevalences of the virus varied significantly between different geographical regions with different epidemiological situation. Most frequently limited number of animals studied and the differences in the sensitivities and specificities of the diagnostic test used could have also influenced on the differences between the studies. No pestivirus has been ever detected in free-ranging reindeer, however, a putative pestivirus strain named V60-Krefeld has been isolated from reindeer kept at a German Zoo in the 1990's. The virus was characterized as border disease virus type 2 (BDV-2) closely related to German ovine strains. The cross-neutralization studies of the semi-domesticated reindeer sera from Sweden suggested infection with a strain related to BDV-1 or BDV-2. The available data indicates that reindeer might be infected by a endemic species-specific BDV-like strain. However, the interspecies transmission of BVDV from domestic animals should not be excluded, since the susceptibility of reindeer to BVDV-1 has been confirmed under experimental conditions.