Global and Phylogenetic Distribution of Quorum Sensing Signals,Acyl Homoserine Lactones,in the Family of Vibrionaceae

Bacterial quorum sensing (QS) and the corresponding signals, acyl homoserine lactones (AHLs), were first described for a luminescent Vibrio species. Since then, detailed knowledge has been gained on the functional level of QS; however, the abundance of AHLs in the family of Vibrionaceae in the environment has remained unclear. Three hundred and one Vibrionaceae strains were collected on a global research cruise and the prevalence and profile of AHL signals in this global collection were determined. AHLs were detected in 32 of the 301 strains using Agrobacterium tumefaciens and Chromobacterium violaceum reporter strains. Ethyl acetate extracts of the cultures were analysed by ultra-high performance liquid chromatography-high resolution mass spectrometry (MS) with automated tandem MS confirmation for AHLs. N-(3-hydroxy-hexanoyl) (OH-C6) and N-(3-hydroxy-decanoyl) (OH-C10) homoserine lactones were the most common AHLs found in 17 and 12 strains, respectively. Several strains produced a diversity of different AHLs, including N-heptanoyl (C7) HL. AHL-producing Vibrionaceae were found in polar, temperate and tropical waters. The AHL profiles correlated with strain phylogeny based on gene sequence homology, however not with geographical location. In conclusion, a wide range of AHL signals are produced by a number of clades in the Vibrionaceae family and these results will allow future investigations of inter- and intra-species interactions within this cosmopolitan family of marine bacteria.     

Accurate Dereplication of Bioactive Secondary Metabolites from Marine-Derived Fungi by UHPLC-DAD-QTOFMS and a MS/HRMS Library

In drug discovery, reliable and fast dereplication of known compounds is essential for identification of novel bioactive compounds. Here, we show an integrated approach using ultra-high performance liquid chromatography-diode array detection-quadrupole time of flight mass spectrometry (UHPLC-DAD-QTOFMS) providing both accurate mass full-scan mass spectrometry (MS) and tandem high resolution MS (MS/HRMS) data. The methodology was demonstrated on compounds from bioactive marine-derived strains of Aspergillus, Penicillium, and Emericellopsis, including small polyketides, non-ribosomal peptides, terpenes, and meroterpenoids. The MS/HRMS data were then searched against an in-house MS/HRMS library of ~1300 compounds for unambiguous identification. The full scan MS data was used for dereplication of compounds not in the MS/HRMS library, combined with ultraviolet/visual (UV/Vis) and MS/HRMS data for faster exclusion of database search results. This led to the identification of four novel isomers of the known anticancer compound, asperphenamate. Except for very low intensity peaks, no false negatives were found using the MS/HRMS approach, which proved to be robust against poor data quality caused by system overload or loss of lock-mass. Only for small polyketides, like patulin, were both retention time and UV/Vis spectra necessary for unambiguous identification. For the ophiobolin family with many structurally similar analogues partly co-eluting, the peaks could be assigned correctly by combining MS/HRMS data and m/z of the [M + Na]⁺ ions.     

A model ensemble approach to determine the humus building efficiency of organic amendments in incubation experiments

Organic amendments are important to sustain soil organic matter (SOM) and soil functions in agricultural soils. Information about the contribution of organic amendments to SOM can be derived from incubation experiments. In this study, data from 72 incubated organic amendments including plant residues, digestates and manure were analysed. The incubation data was compiled from three experimental setups with varying incubation times, soils and incubation temperatures, in which CO₂ release was measured continuously. The analysis of the incubation data was performed with an approach relying on conceptual parts of C-TOOL, CCB, Century, ICBM, RothC and Yasso which are all well-approved first-order carbon models that differ in structure and abstraction level. All models are an approximation of reality, whereby each model differs in understanding of the processes involved in soil carbon dynamics. To accumulate the advantages from each model a model ensemble was performed for each substrate. With the ability of each carbon model to compute the distribution of carbon into specific SOM pools a new approach for evaluating organic amendments in terms of humus building efficiency is presented that, depends on the weighted model fit of each ensemble member. Depending on the organic substrate added to the soil, the time course of CO₂ release in the incubation studies was predicted with different accuracy by the individual model concepts. Averaging the output of the individual models leads to more robust prediction of SOM dynamics. The E_HUM value is easy to interpret and the results are in accordance with the literature.     

The occurrence of summer mastitis in Jutland (Denmark) in relation to meteorological factors

Data for incidence of summer mastitis in 14 veterinary practices in Jutland (Denmark) were related to meteorological factors using statistical methods. A correlation and regression analysis showed that summer mastitis occurred earlier in years with high temperature and precipitation in early summer. The annual number of cases markedly increased over the years 1953 to 1980. This increase was described by applying a linear increase with with year to a logarithmic transformation of the annual incidence.Analysis of 10-day incidences from the veterinary practices and daily incidence from a test site at St. Vildmose (Jutland) showed that incidence of summer mastitis could be related to temperature and wind velocity within the previous 10 days. High temperature and low wind velocity in this period increase the incidence. This indicates that the incubation period of summer mastitis is at maximum about 10 days. The activity of the fly (Hydrotaea irritans Fall), which acts as mechanical vector for the bacteria is influenced by temperature and wind velocity. This may explain the variation in incidence of summer mastitis during the season and from year to year.     

Effects of breeding season length and calving season on range beef cow productivity

A 5-yr study was conducted beginning in 1983 with 460 cows to evaluate the effects of three breeding seasons (30, 45, and 70 d in length) and two times of spring calving, March (early) and April (late), on cattle production under Nebraska Sandhills range conditions. Criteria evaluated included pregnancy and weaning percentages, calving date and distribution, cow weights and body condition at four intervals, calf birth and weaning weights, and cow productivity. The 30-d breeding season included a 10-d estrus synchronization and AI period; in the other breeding seasons only natural breeding was used. The same sires were used over the entire study period. Percentage of cows pregnant and percentage of calves weaned were lower (P less than .01) for cows bred for 30 d than for cows bred for 45 or 70 d. Average calving dates were similar among the breeding groups within the early and late calving herds. Pregnancy rates from AI were higher (P less than .01) for the cows calving in April (64%) than for the cows calving in March (41%). Cows calving in April lost less weight between precalving and prebreeding and were heavier (P less than .05) at prebreeding time than the cows calving in March. Calf weaning weights were not different (P greater than .10) among any of the breeding season groups or between the two calving herds when calves were weaned at a similar age. Cow productivity (calf weaning weight per breeding female) was highest (P less than .05) for the cows bred for 70 d (186 kg), intermediate for the cows bred for 45 d (172 kg), and lowest for cows bred for 30 d (162 kg).(ABSTRACT TRUNCATED AT 250 WORDS)     

The seal death in Danish waters 1988. 2. Virological studies.

Mass abortions and high mortality were observed in harbour seals in Danish waters during 1988. Severe pneumonia and emphysema were typical clinical and post-mortem findings. Virological studies were carried out to identify the cause of the epidemic. Although seal herpesvirus (SeHV) was isolated in 23 of 114 animals this virus was subsequently found not to be the primary cause of the disease. Following the observation of seroconversion against canine distemper virus (CDV) in diseased seals (Osterhaus & Vedder 1988) a CDV-like morbillivirus (phocine distemper virus, PDV) was identified in organs of diseased animals. It is concluded that the epidemic was caused by introduction of PDV into a highly susceptible population presumably free from morbillivirus infection. The origin of PDV remains unknown but evidence of prior morbillivirus infection has been found in arctic and antarctic seal populations.     

Ecosystem health as a clinical rotation for senior students in Canadian veterinary schools.

We describe 4 years of an experimental rotation in ecosystem health offered to senior veterinary students in Canada. Faculty from the 4 Canadian veterinary colleges collaborated in offering the rotation once annually at 1 of the colleges. The 1st rotation was held in Guelph in 1993, followed in successive years by rotations at Saskatoon, Saint-Hyacinthe, and Charlottetown. The rotation is a predominantly field-based experience that allows students to work with veterinary and other role models who are actively engaged in clinical research related to ecosystem health. Five specific field studies that worked particularly well during the rotations are presented. These studies involved investigating mortality in wildlife due to botulism, designing an environmental surveillance system around herds of beef cattle, using belugas to evaluate the health of the St. Lawrence River, dealing with competition for water use by aquaculture and agriculture, and exploring the role of veterinarians during major coastal oil spills. The experience has resulted in our developing the subject matter, field examples, teaching approach, and confidence necessary to make ecosystem health the focus of a productive clinical rotation for senior year veterinary students.     

Benefit-cost analysis of the current African swine fever eradication program in Spain and of an accelerated program

We predicted the rate of spread and economic losses due to African swine fever (ASF) infections in Spanish swine for a 20 year period using current epidemiological data. Two possibilities for eradication were examined: continuation of the eradication program presently in use, assuming continued funding at present levels, and an accelerated eradication program using more frequent testing to identify seropositive carrier animals. Both programs theoretically would result in the eradication of African swine fever. The calculated benefit-cost ratios were 1.23 and 1.47 respectively. A reduction in current yearly funding would result in a benefit-cost ratio of 0.97, making the program unprofitable according to our model.

The projected year for the eradication of outbreaks is 1996 using provinces as units in the model. Similarly, the projected year for the eradication of seropositive animals is 2001.     

Variance Components of an Enzyme‐linked Immunosorbent Assay for Detection of IgG Antibodies in Milk Samples to Mycobacterium avium subspecies paratuberculosis in Dairy Cattle

Milk samples from 120 cows were tested up to 10 times in an enzyme‐linked immunosorbent assay (ELISA) for detection of antibodies to Mycobacterium avium ssp. paratuberculosis. The purpose of the study was to estimate variance components of the assay attributable to laboratory factors using mixed model theory. Because of significant interaction between the between‐run, between‐day and between‐plate variables, the ELISA‐plate variable was nested in run‐number and run‐number was nested in day‐number. The nested variable accounted for 68% of the laboratory variability (P<0.001), whereas the intra‐plate variability accounted for only 0.04% of the laboratory variability (P>0.9). Therefore, it was concluded that the intra‐plate variability could be ignored whereas the variability from the combined run‐day‐plate variable should be considered in any analyses based on the ELISA.