Zoonotic poxviruses

Poxviruses compromise a group of long known important pathogens including some zoonotic members affecting lifestock animals and humans. While whole genome sequence analysis started to shed light into the molecular mechanisms underlying host cell infection, viral replication as well as virulence, our understanding of poxvirus maintenance in nature and their transmission to humans is still poor. During the last two decades, reports on emerging human monkeypox outbreaks in Africa and North America, the increasing number of cowpox virus infections in cats, exotic animals and humans and cases of vaccinia virus infections in humans in South America and India reminded us that – beside the eradicated smallpox virus – there are other poxviruses that can cause harm to men. We start to learn that the host range of some poxviruses is way broader than initially thought and that mainly rodents seem to function as virus reservoir. The following review is aiming to provide an up-to-date overview on the epidemiology of zoonotic poxviruses, emphasizing orthopoxviruses. By outlining the current knowledge of poxvirus transmission, we hope to raise the awareness about modes of acquisition of infections and their proper diagnosis.     

Fusarium oxysporum f. sp. radicis‐vanillae is the causal agent of root and stem rot of vanilla

Root and stem rot (RSR) is a very detrimental disease of vanilla worldwide. Fusarium oxysporum is frequently associated with the disease but other Fusarium species are also reported. In this international study, 52 vanilla plots were surveyed in three of the most important vanilla producing countries (Madagascar, Reunion Island and French Polynesia) in order to determine the aetiology of RSR disease. Subsets from the 377 single‐spored Fusarium isolates recovered from rotten roots and stems in the surveys were characterized by molecular genotyping (EF1α and IGS gene sequences) and pathogenicity assays on Vanilla planifolia and V. ×tahitensis, the two commercially grown vanilla species. Fusarium oxysporum was shown to be the principal species responsible for the disease, representing 79% of the isolates recovered from the RSR tissues, 40% of which induced severe symptoms on inoculated plantlets. Fusarium oxysporum isolates were highly polyphyletic regardless of geographic origin or pathogenicity. Fusarium solani, found in 15% of the samples and inducing only mild symptoms on plantlets, was considered a secondary pathogen of vanilla. Three additional Fusarium species were occasionally isolated in the study (F. proliferatum, F. concentricum and F. mangiferae) but were nonpathogenic. Histopathological preparations observed in wide field and multiphoton microscopy showed that F. oxysporum penetrated the root hair region of roots, then invaded the cortical cells where it induced necrosis in both V. planifolia and V. ×tahitensis. The hyphae never invaded the root vascular system up to 9 days post‐inoculation. As a whole, the data demonstrated that RSR of vanilla is present worldwide and that its causal agent should be named F. oxysporum f. sp. radicis‐vanillae.     

Die Entwicklung des Obstbaus in Südtirol: I. Die landwirtschaftliche Aus- und Weiterbildung

South Tyrol (northern Italy) harbours the largest continuous apple producing area in Europe, with an annual fruit harvest of around one million of metric tonnes. Every tenth apple produced in EU-28 originates from South Tyrol and this fruit thus represents the most important export product of this region, which contributes to 55 % of the added value from agriculture. The success of apple production depends on several key factors, one of which is education and professional training in agriculture and fruit production, and this topic is elaborated in greater detail in the present work. In 1874, the ‘Agricultural Institute and Experimental Station’ in S. Michele upon Etsch/Adige was inaugurated as an institution for higher agricultural education and research for the entire territory of the historical Tyrol. The first vocational school for agriculture in South Tyrol was founded in 1909 in Sterzing/Vipiteno (later transferred to Dietenheim/Teodone nearby Bruneck/Brunico), but soon, with World War I, the agricultural education in German language collapsed completely and was re-established only after World War II. The work of the vocational school in Dietenheim/Teodone was pursued, and in 1952 opened the Vocational School for Agriculture and Forestry Fürstenburg, in 1962 the Vocational School for Fruit, Viticulture and Horticulture Laimburg and in 1978 the Vocational School for Agriculture and Domestic Economy Salern. Increasing student numbers demonstrate the success of these schools. In 1980, finally, the High School for Agriculture in Auer/Ora nearby Bozen/Bolzano was founded and in 1999 started the Bachelor’s study programme in Agricu­ltural Technology and Economics at the Free University of Bozen/Bolzano. Nowadays, the agricultural education in South Tyrol is covered by a full range of school types ranging from vocational schools to university level doctoral studies.     

The inhibiting effect of nitrate fertilisation on methane uptake of a temperate forest soil is influenced by labile carbon

Upland soils are the most important terrestrial sink for the greenhouse gas CH₄. The oxidation of CH₄ is highly influenced by reactive N which is increasingly added to many ecosystems by atmospheric deposition and thereby also alters the labile C pool in the soils. The interacting effects of soil N availability and the labile C pool on CH₄ oxidation are not well understood. We conducted a laboratory experiment with soil columns consisting of homogenised topsoil material from a temperate broad-leaved forest to study the net CH₄ flux under the combined or isolated addition of NO ₃ ^? and glucose as a labile C source. Addition of NO ₃ ^? and glucose reduced the net CH₄ uptake of the soil by 86% and 83%, respectively. The combined addition of both agents led to a nearly complete inhibition of CH₄ uptake (reduction by 99.4%). Our study demonstrates a close link between the availability of C and N and the rate of CH₄ oxidation in temperate forest soils. Continued deposition of NO ₃ ^? has the potential to reduce the sink strength of temperate forest soils for CH₄.     

Spatial variability of nitrous oxide emissions in an unmanaged old‐growth beech forest

Nitrous oxide (N₂O) is a high‐impact greenhouse gas. Due to the scarcity of unmanaged forests in Central Europe, its long‐term natural background emission level is not entirely clear. We measured soil N₂O emissions in an unmanaged, old‐growth beech forest in the Hainich National Park, Germany, at 15 plots over a 1‐year period. The average annual measured N₂O flux rate was (0.49 ± 0.44) kg N ha^–1 y^–1. The N₂O emissions showed background‐emission patterns with two N₂O peaks. A correlation analysis shows that the distance between plots (up to 380 m) does not control flux correlations. Comparison of measured data with annual N₂O flux rates obtained from a standard model (Forest‐DNDC) without site‐specific recalibration reveals that the model overestimates the actual measured N₂O flux rates mainly in spring. Temporal variability of measured N₂O flux was better depicted by the model at plots with high soil organic C (SOC) content. Modeled N₂O flux rates were increased during freezing only when SOC was > 0.06 kg C kg^–1. The results indicate that the natural background of N₂O emissions may be lower than assumed by most approaches.     

Effect of mineral nitrogen fertilizer forms on N2O emissions from arable soils in winter wheat production

Nitrogen fertilizers are supposed to be a major source of nitrous oxide (N₂O) emissions from arable soils. The objective of this study was to compare the effect of N forms on N₂O emissions from arable fields cropped with winter wheat (Triticum aestivum L.). In three field trials in North‐West Germany (two trials in 2011/2012, one trial in 2012/2013), direct N₂O emissions during a one‐year measurement period, starting after application of either urea, ammonium sulfate (AS) or calcium ammonium nitrate (CAN), were compared at an application rate of 220 kg N ha^?1. During the growth season (March to August) of winter wheat, N₂O emission rates were significantly higher in all three field experiments and in all treatments receiving N fertilizer than from the non‐fertilized treatments (control). At two of the three sites, cumulative N₂O emissions from N fertilizer decreased in the order of urea > AS > CAN, with emissions ranging from 522–617 g N ha^?1 (0.24–0.28% of applied fertilizer) for urea, 368–554 g N ha^?1 (0.17–0.25%) for AS, and 242–264 g N ha^?1 (0.11–0.12%) for CAN during March to August. These results suggest that mineral nitrogen forms can differ in N₂O emissions during the growth period of winter wheat. Strong variations in the seasonal dynamics of N₂O emissions between sites were observed which could partly be related to weather events (e.g., precipitation). Between harvest and the following spring (post‐harvest period) no significant differences in N₂O emissions between fertilized and non‐fertilized treatments were detected on two of three fields. Only on one site post‐harvest emissions from the AS treatment were significantly higher than all other fertilizer forms as well as compared to the control treatment. The cumulative one‐year emissions varied depending on fertilizer form across the three field sites from 0.05% to 0.51% with one exception at one field site (AS: 0.94%). The calculated overall fertilizer induced emission averaged for the three fields was 0.38% which was only about 1/3 of the IPCC default value of 1.0%.     

Stability of vaccinia-vectored recombinant oral rabies vaccine under field conditions: a 3-year study

Rabies is an incurable zoonotic disease caused by rabies virus, a member of the rhabdovirus family. It is transmitted through the bite of an infected animal. Control methods, including oral rabies vaccination (ORV) programs, have led to a reduction in the spread and prevalence of the disease in wildlife. This study evaluated the stability of RABORAL, a recombinant vaccinia virus vaccine that is used in oral rabies vaccination programs. The vaccine was studied in various field microenvironments in order to describe its viability and facilitate effective baiting strategies. Field microenvironments influenced the stability of this vaccine in this study. This study emphasizes the importance of understanding how vaccines perform under varying field conditions in order to plan effective baiting strategies.     

Analysis of bacterial load and prevalence of mixed infections with Lawsonia intracellularis, Brachyspira hyodysenteriae and/or Brachyspira pilosicoli in German pigs with diarrhoea

Lawsonia (L.) intracellularis, Brachyspira (B.) hyodysenteriae and B. pilosicoli are important pathogens in domestic pig production world-wide, responsible for porcine intestinal adenomatosis, swine dysentery, and porcine intestinal spirochetosis, respectively. Conventional PCR is the major diagnostic tool in the detection of the three pathogens, but the sole detection of bacterial DNA might lead to misinterpretations of results with respect to their clinical relevance, especially with mixed infections. Thus, the present study targeted the detection and quantification of the three pathogens in samples from herds with a case history of diarrhoea. Herds and samples were selected by the practitioners on a voluntary basis. Results were based on 1176 individual samples from 95 herds from Southern Germany. The pathogens were detected simultaneously by multiplex real-time PCR. The overall prevalence for L. intracellularis, B. hyodysenteriae and B. pilosicoli was 12.6%, 8.4% and 3.2% in faecal samples and 48.4%, 24.2% and 31.6% in herds, respectively. Sixty one percent, 82.6%, and 73.4% of herds positive for L. intracellularis, B. hyodysenteriae, and B. pilosicoli, respectively, had mixed infections. Median log values of DNA equivalents/g of faeces for L. intracellularis, B. hyodysenteriae and B. pilosicoli were 3.3, 5.9 and 3.2, with maxima of 8.3, 8.0 and 6.3, respectively. Within herd prevalence of B. hyodysenteriae and B. pilosicoli as well as the load of B. hyodysenteriae were significantly associated with the severity of diarrhoea.     

Optimising magnetic resonance image quality of the ear in healthy dogs

The aim of this study was to develop an examination protocol for magnetic resonance imaging, in order to display diagnostically important information of the canine middle and inner ear. To ensure that this protocol could also be used as a basis for determining pathological changes, the anatomical structures of the ear were presented in detail. To minimise stress through anaesthesia in live animals, preliminary examinations were carried out on four dog cadavers. During these initial examinations, three-dimensional (3D) sequences proved to be superior to two-dimensional ones. Therefore, only 3D sequences were applied for the main examinations performed on six clinically healthy Beagles. The anonymised MR images were rated by three experienced reviewers using a five-point scale. The most valuable sequence was a T2-weighted CISS sequence (TR = 16.7 ms, TE = 8.08 ms). This sequence proved to be most suitable for illustrating the inner ear structures and enabled good tissue contrasts. The sequence ranked second best was also a T2-weighted DESS sequence (TR = 19 ms, TE = 6 ms), allowing the imaging of the tympanic cavity and enabling 3D reconstruction due to its isotropic voxels. Due to low contrast and strong noise, the other sequences (TSE, FISP, MP RAGE) were not suitable for anatomical illustration of the middle and inner ear.     

Genotyping of bovine Prototheca mastitis isolates from Poland

Bovine mastitis due to unicellular, achlorophyllous algae of the genus Prototheca is a serious and complex ailment that accounts for high economic losses in the dairy industry. Bovine protothecal mastitis has been almost exclusively associated with only one species Prototheca zopfii. Recently, based on the 18S rDNA sequence analysis, P. zopfii has been divided into three genotypes (1-3), of which P. zopfii genotype 3 has been given the status of a new species Prototheca blaschkeae sp. nov. The aim of this study was to investigate the genotypic composition of the population of P. zopfii bovine mastitis isolates from Poland, according to that threefold classification. Of the 44 isolates tested, 43 (98%) were identified as P. zopfii genotype 2. Only one isolate was assigned to the newly established P. blaschkeae species. These findings clearly show the predominance of the P. zopfii genotype 2 in the etiology of bovine mammary protothecosis in Poland, but also underline the involvement of P. blaschkeae in the disease. The study is the first to provide molecular characterization of Polish P. zopfii mastitis isolates, collected from different regions of Poland.