Animal health in the 21st century-a global challenge

On the occasion of the centenary of the Friedrich-Loeffler-Institut, a conference entitled 'Animal Health in the 21st Century' was held in Greifswald, Germany, on 11-13 October 2010 to discuss current and future challenges regarding the global situation regarding infectious animal diseases and zoonoses, animal breeding, animal nutrition and animal welfare. Particular attention was paid to the impact of recent developments and anticipated future trends on livestock production.     

Isolation and characterisation of equine influenza viruses (H3N8) from Europe and North America from 2008 to 2009

Like other influenza A viruses, equine influenza virus undergoes antigenic drift. It is therefore essential that surveillance is carried out to ensure that recommended strains for inclusion in vaccines are kept up to date. Here we report antigenic and genetic characterisation carried out on equine influenza virus strains isolated in North America and Europe over a 2-year period from 2008 to 2009. Nasopharyngeal swabs were taken from equines showing acute clinical signs and submitted to diagnostic laboratories for testing and virus isolation in eggs. The sequence of the HA1 portion of the viral haemagglutinin was determined for each strain. Where possible, sequence was determined directly from swab material as well as from virus isolated in eggs. In Europe, 20 viruses were isolated from 15 sporadic outbreaks and 5 viruses were isolated from North America. All of the European and North American viruses were characterised as members of the Florida sublineage, with similarity to A/eq/Lincolnshire/1/07 (clade 1) or A/eq/Richmond/1/07 (clade 2). Antigenic characterisation by haemagglutination inhibition assay indicated that the two clades could be readily distinguished and there were also at least seven amino acid differences between them. The selection of vaccine strains for 2010 by the expert surveillance panel have taken these differences into account and it is now recommended that representatives of both Florida clade 1 and clade 2 are included in vaccines.     

Carry-over of thermophilic Campylobacter spp. between sequential and adjacent poultry flocks

Nineteen flocks of four poultry species were monitored at a veterinary field station to investigate the distribution and spread of Campylobacter genotypes between sequential and adjacent flocks. Caecal and liver samples were obtained at frequent intervals from birds of all flocks and examined for Campylobacter. Amplified fragment length polymorphism (AFLP) analysis was performed to genotype Campylobacter isolates. Of the 1643 caecal and liver samples investigated, 452 (27.5%) caecal samples and 11 (0.7%) liver samples contained Campylobacter. Of the caecal isolates 76.3% were identified as Campylobacter jejuni and 23.7% were identified as Campylobacter coli. Poultry flocks were largely colonized by more than one AFLP type and an intense exchange of Campylobacter genotypes between different poultry flocks occurred. These findings indicate that multiple genotypes can constitute the Campylobacter population within single poultry flocks, hinting to different sources of exposure and/or genetic drifts within the Campylobacter population. Nevertheless, in most flocks single Campylobacter genotypes predominated. Some strains superseded others resulting in colonization by successive Campylobacter genotypes during the observation period. In conclusion, the data demonstrate that the large genetic diversity of Campylobacter must be considered in epidemiological evaluations and microbial risk assessments of Campylobacter in poultry.     

Genetic diversity of diploid Japanese strawberry species based on microsatellite markers

The United States Department of Agriculture (USDA)—Agricultural Research Service (ARS)—National Clonal Germplasm Repository (NCGR) in Corvallis, Oregon, is a genebank that preserves strawberry genetic resources. Representatives of two Japanese diploid species, Fragaria iinumae Makino and F. nipponica Makino were collected for conservation by the NCGR during an expedition to Hokkaido, Japan. Fragaria iinumae may be a genome contributor to the cultivated octoploid strawberries. The objective of this study was to evaluate the genetic diversity of these two species by using simple sequence repeat (SSR) markers. Twenty of 82 Fragaria-derived SSRs, polymorphic among and within the two species, were selected for genetic analysis of 137 accessions. Genetic diversity, based on the proportion of shared alleles between the two species, in F. nipponica (0.4542) and F. iinumae (0.1808) was significantly different. Three wild interspecific hybrids were identified from intermediate memberships in the two diploid species groups revealed by using the clustering program, Structure. Principal coordinate analysis followed by non-parametric modal clustering (PCO-MC) grouped accessions into two clusters representing the two diploid species. Further clustering within the species groups generated with the program, STRUCTURAMA^TM, resulted in seven subclusters in F. iinumae and three in F. nipponica, which may represent breeding populations appropriate for clonal conservation. Long-term preservation of the species populations and the limited number of hybrids on the island is discussed relative to their geographical distribution and the geological history of Hokkaido Island.     

Systematic studies on the chemical structure and umami enhancing activity of Maillard-modified guanosine 5'-monophosphates

Recent investigations on taste active principles in nucleotide rich yeast extracts led to the discovery of (R)- and (S)-N(2)-(1-carboxyethyl)-guanosine 5'-monophosphate as previously not reported umami enhancing molecules formed upon the Maillard reaction of guanosine 5'-monophosphate (5'-GMP) with dihydroxyacetone and glyceraldehyde, respectively. In the present study, systematic Maillard-type model reactions were performed with 5'-GMP and a homologous series of monosaccharides exhibiting a C(3)- to C(6)-carbon skeleton as well as with the reducing disaccharide maltose in the presence of an amino acid. By preparative RP-HPLC, various (R)- and (S)-N(2)-(1-carboxyalkyl)-guanosine 5'-monophosphates and (R)- and (S)-N(2)-(1-alkylamino)carbonylalkyl)guanosine 5'-monophosphates were isolated and identified by means of LC-MS, LC-TOF-MS, and 1D/2D-NMR spectroscopy. Sensory evaluation of these Maillard products revealed β-values for umami enhancement between 0.06 and 7.0 and identified a strong influence of the stereochemistry as well as the chain length of the N(2)-substituent on the umami enhancing activity. For all of the compounds evaluated, the (S)-configured isomers showed higher taste impact, whereas the (R)-isomers showed only marginal β-values, thus underlining the stereospecifity of the umami taste receptor binding site.     

Investigation of the heating rate dependency associated with the loss of crystalline structure in sucrose, glucose, and fructose using a thermal analysis approach (part I)

Thermodynamic melting occurs at a single, time-independent temperature with a constant enthalpy value. However, substantial variation in the melting parameters (T(m onset), T(m peak), and ΔH) for sucrose, glucose, and fructose has been reported in the literature. Although a number of explanations have been put forth, they do not completely account for the observed variation. Thus, this research was performed to elucidate the fundamental mechanism underlying the loss of crystalline structure in the sugars using both thermal (Part I) and chemical (Part II) analysis approaches. A strong heating rate dependency observed in the melting parameters for the sugars implies the occurrence of a kinetic process during the loss of crystalline structure. The difference in heat capacity and modulated heat flow amplitude in the stepwise quasi-isothermal modulated differential scanning calorimetry experiments for the sugars compared to indium and mannitol (thermodynamic melting comparison materials) strongly suggests thermal decomposition as the kinetic process responsible for the loss of crystalline structure, which is the critical difference between our conclusion and others. We propose the term "apparent melting" to distinguish the loss of crystalline structure due to a kinetic process, such as thermal decomposition, from thermodynamic melting.     

Antifungal activity and fungal metabolism of steroidal glycosides of Easter lily (Lilium longiflorum Thunb.) by the plant pathogenic fungus, Botrytis cinerea

Botrytis cinerea Pers. Fr. is a plant pathogenic fungus and the causal organism of blossom blight of Easter lily (Lilium longiflorum Thunb.). Easter lily is a rich source of steroidal glycosides, compounds which may play a role in the plant-pathogen interaction of Easter lily. Five steroidal glycosides, including two steroidal glycoalkaloids and three furostanol saponins, were isolated from L. longiflorum and evaluated for fungal growth inhibition activity against B. cinerea, using an in vitro plate assay. All of the compounds showed fungal growth inhibition activity; however, the natural acetylation of C-6' of the terminal glucose in the steroidal glycoalkaloid, (22R,25R)-spirosol-5-en-3β-yl O-α-L-rhamnopyranosyl-(1→2)-[6-O-acetyl-β-D-glucopyranosyl-(1→4)]-β-D-glucopyranoside (2), increased antifungal activity by inhibiting the rate of metabolism of the compound by B. cinerea. Acetylation of the glycoalkaloid may be a plant defense response to the evolution of detoxifying mechanisms by the pathogen. The biotransformation of the steroidal glycoalkaloids by B. cinerea led to the isolation and characterization of several fungal metabolites. The fungal metabolites that were generated in the model system were also identified in Easter lily tissues infected with the fungus by LC-MS. In addition, a steroidal glycoalkaloid, (22R,25R)-spirosol-5-en-3β-yl O-α-L-rhamnopyranosyl-(1→2)-β-D-glucopyranoside (6), was identified as both a fungal metabolite of the steroidal glycoalkaloids and as a natural product in L. longiflorum for the first time.