Analysis of moniliformin in maize plants using hydrophilic interaction chromatography

A novel HPLC method was developed for detection of the Fusarium mycotoxin, moniliformin in whole maize plants. The method is based on hydrophilic interaction chromatography (HILIC) on a ZIC zwitterion column combined with diode array detection and negative electrospray mass spectrometry (ESI(-)-MS). Samples were extracted using acetonitrile-water (85:15), and the extracts were cleaned up on strong anion exchange columns. By this procedure we obtained a recovery rate of 57-74% moniliformin with a limit of detection at 48 ng/g and a limit of quantification at 96 ng/g using UV detection at 229 nm, which is comparable to current methods used. Limit of detection and quantification using ESI(-)-MS detection was 1 and 12 ng/g, respectively. Screening of maize samples infected with the moniliformin producing fungi F. avenaceum, F. tricinctum, or F. subglutinans detected moniliformin levels of 1-12 ng/g in 15 of 28 samples using ESI(-)-MS detection. To our knowledge this is the first example of HILIC separation in mycotoxin analysis.     

After nasopharyngeal infection,foot-and-mouth disease virus serotype A RNA is shed in bovine milk without associated mastitis

Foot-and-mouth disease (FMD) is a highly contagious aphthoviral infection of cloven-hoofed animals, inducing vesiculopustular stomatitis, pododermatitis, and thelitis. Vesicular fluid represents a major pathway of virus excretion, but bovine milk is another important source of virus shedding. We describe here the time course of FMD virus (FMDV) excretion in the milk and characterize associated lesions in the mammary gland. Three dairy cows were infected by nasopharyngeal instillation of FMDV and monitored over 12 d. Autopsy was performed at the end of the study, and specimens were collected for histopathology, IHC, and RT-qPCR. All 3 cows developed fever, drooling, vesiculopustular stomatitis, interdigital dermatitis, and thelitis. FMDV RNA was detectable in whole milk until the end of the trial, but only transiently in saliva, nasal secretions, and blood serum. Although histology confirmed vesiculopustular lesions in the oral and epidermal specimens, the mammary glands did not have unequivocal evidence of FMDV-induced inflammation. FMDV antigen was detectable in skin and oral mucosa, but not in the mammary gland, and FMDV RNA was detectable in 9 of 29 samples of squamous epithelia but only in 1 of 12 samples of mammary gland.     

The apparent prevalence of skin lesions suspected to be human-inflicted in Danish finishing pigs at slaughter

Skin lesions on pigs inflicted by humans compromise animal welfare and are the subject of increased public and political attention in Denmark. Systematic surveillance of such skin lesions was enforced in April 2010 at all Danish pig abattoirs, through the recording of meat inspection Code 904 for the presence of skin lesions suspected to be human inflicted. The objectives of the present study were to (a) estimate the apparent prevalence of Code 904s at the pig and herd owner level; (b) characterise the distribution of deliveries with pigs demonstrating a Code 904; (c) characterise the distribution of herd owners with repeated Code 904 recordings; and (d) determine the developments in Code 904 prevalence over time in Danish finishing pigs in the period from May 1, 2010 to September 30, 2013.     

Prevalence and risk factors of Coxiella burnetii seropositivity in Danish beef and dairy cattle at slaughter adjusted for test uncertainty

Antibodies to Coxiella burnetii have been found in the Danish dairy cattle population with high levels of herd and within herd seroprevalences. However, the prevalence of antibodies to C. burnetii in Danish beef cattle remains unknown. The objectives of this study were to (1) estimate the prevalence and (2) identify risk factors associated with C. burnetii seropositivity in Danish beef and dairy cattle based on sampling at slaughter.     

Herd diagnosis of low pathogen diarrhoea in growing pigs – a pilot study

Background

The major indication for antibiotic use in Danish pigs is treatment of intestinal diseases post weaning. Clinical decisions on antibiotic batch medication are often based on inspection of diarrhoeic pools on the pen floor. In some of these treated diarrhoea outbreaks, intestinal pathogens can only be demonstrated in a small number of pigs within the treated group (low pathogen diarrhoea). Termination of antibiotic batch medication in herds suffering from such diarrhoea could potentially reduce the consumption of antibiotics in the pig industry. The objective of the present pilot study was to suggest criteria for herd diagnosis of low pathogen diarrhoea in growing pigs.Data previously collected from 20 Danish herds were used to create a case series of clinical diarrhoea outbreaks normally subjected to antibiotic treatment. In the present study, these diarrhoea outbreaks were classified as low pathogen (<15% of the pigs having bacterial intestinal disease) (n =5 outbreaks) or high pathogen (≥15% of the pigs having bacterial intestinal disease) (n =15 outbreaks). Based on the case series, different diagnostic procedures were explored, and criteria for herd diagnosis of low pathogen diarrhoea were suggested. The effect of sampling variation was explored by simulation.

Results

The diagnostic procedure with the highest combined herd-level sensitivity and specificity was qPCR testing of a pooled sample containing 20 randomly selected faecal samples. The criteria for a positive test result (high pathogen diarrhoea outbreak) were an average of 1.5 diarrhoeic faecal pools on the floor of each pen in the room under investigation and a pathogenic bacterial load ≥35,000 per gram in the faecal pool tested by qPCR. The bacterial load was the sum of Lawsonia intracellularis, Brachyspira pilosicoli and Escherichia coli F4 and F18 bacteria per gram faeces. The herd-diagnostic performance was (herd-level) diagnostic sensitivity =0.99, diagnostic specificity =0.80, positive predictive value =0.94 and negative predictive value =0.96.

Conclusions

The pilot study suggests criteria for herd diagnosis of low pathogen diarrhoea in growing pigs. The suggested criteria should now be evaluated, and the effect of terminating antibiotic batch medication in herds identified as suffering from low pathogen diarrhoea should be explored.

Electronic supplementary material

The online version of this article (doi:10.1186/2046-0481-67-24) contains supplementary material, which is available to authorized users.     

When fish shoals meet: outcomes for evolution and fisheries

The mechanisms underlying the social structure of free‐ranging fish shoals have received little attention in comparison to functional studies on shoaling. Recently, however, a number of investigations, both in the marine and in the freshwater environment, have begun to address the underlying mechanisms by concentrating on interactions between free‐ranging shoals. The rates of shoal encounters can influence the opportunities for individual assortment by phenotype and selection of shoal size; act as a constraint on the observed patterns of shoal structure by restricting individual choice behaviour and were found to be high in several small freshwater species (where intershoal distances were small), but lower in marine species where shoals were more dispersed. The duration of encounters may play a role in that it affects both the time available for assessment (of the encountered shoal) as well as that for exchange of individuals. Scarce published information on the outcome of shoal encounters suggests that the outcome of shoal encounters is influenced by shoal composition but not by shoal size. Individual behaviour may have evolved to maintain shoal size when shoals encounter, but when fish populations are depleted by fishing, this trait can exacerbate range and stock collapse. Furthermore, an understanding of the dynamics of shoal encounters has important consequences for the evolution of reciprocal altruism and the transmission of information through social learning within populations. Finally, information on encounter rates between shoals and the number of individuals that are exchanged on such occasions could be important for making predictions about the spread of disease through fish populations.     

Marennine,Promising Blue Pigments from a Widespread Haslea Diatom Species Complex

In diatoms, the main photosynthetic pigments are chlorophylls a and c, fucoxanthin, diadinoxanthin and diatoxanthin. The marine pennate diatom Haslea ostrearia has long been known for producing, in addition to these generic pigments, a water-soluble blue pigment, marennine. This pigment, responsible for the greening of oysters in western France, presents different biological activities: allelopathic, antioxidant, antibacterial, antiviral, and growth-inhibiting. A method to extract and purify marennine has been developed, but its chemical structure could hitherto not be resolved. For decades, H. ostrearia was the only organism known to produce marennine, and can be found worldwide. Our knowledge about H. ostrearia-like diatom biodiversity has recently been extended with the discovery of several new species of blue diatoms, the recently described H. karadagensis, H. silbo sp. inedit. and H. provincialis sp. inedit. These blue diatoms produce different marennine-like pigments, which belong to the same chemical family and present similar biological activities. Aside from being a potential source of natural blue pigments, H. ostrearia-like diatoms thus present a commercial potential for aquaculture, cosmetics, food and health industries.     

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.     

Biomass equations for seven different tree species growing in coppice-with-standards forests in Central Germany

With an increasing demand of sustainable raw materials for bioenergy use, coppicing as management approach to increase the biomass production of forests is becoming of greater importance. This study describes the parameterization of biomass equations for six tree species traditionally used in coppices forests, namely sycamore maple (Acer pseudoplatanus L.), field maple (Acer campestre L.), European ash (Fraxinus excelsior L.), European hornbeam (Carpinus betulus L.), downy birch (Betula pubescens Ehrh.), and common hazel (Corylus avellana L.) growing in coppice-with-standard systems in Lower Saxony, Germany. The parameterization was based on measurements of over 950 trees sampled from two forest sites. The sampled trees were felled and separated into three biomass compartments (stem, coarse branches, and fine brushwood) and weighed on site. The dry weight of sub samples from each compartment was measured. Equations were derived for total aboveground biomass, stem biomass, and crown biomass using regression analyses. We either used diameter at breast height as single independent explanatory variable or in combination with tree height. Biomass production of stump sprouts and generatively grown stems was compared for ash and sycamore maple. In the same age classes, it was found that ash stump sprouts had a slightly higher production than seed-grown stems. For sycamore maple, no difference was detected.     

Allele distributions of two novel SNPs within the sheep Cyp19 gene

Oestrogen is an important regulator of reproduction and growth. The key enzyme of oestrogen biosynthesis, aromatase cytochrome P450, is encoded by the Cyp19 gene. In order to generate genetic markers for the sheep Cyp19 gene, two novel single nucleotide polymorphisms (SNPs), one located in promoter 2 (P2), the other one in intron 9 (I9), were identified by a comparative sequencing approach. The allele distributions of both SNPs were investigated by means of Polymerase chain reaction‐restriction fragment length polymorphism analysis (PCR‐RFLP) in five economically relevant sheep breeds (British Milk Sheep, Carranzana, Latxa Black Face, Latxa White Face, Merino) and three ancient Hungarian breeds kept as gene reserves (Cikta, Racka, Tsigai). In British Milk Sheep, only the intronic SNP was present whereas in Merino, Cikta, Racka, Tsigai, Carranzana, Latxa Black Face and Latxa White Face, both SNPs could be found. This indicates that the newly identified SNPs can be used as markers for the Cyp19 locus in various sheep breeds.