Introduction and comparison of two methods of assessment of coronary lesions in Atlantic salmon, Salmo salar L

This study investigated and compared quantitative and semi-quantitative coronary lesion evaluation in Atlantic salmon, Salmo salar. A total of 121 immature farmed and 47 sexually mature wild Atlantic salmon were included. Coronary arteries from all fish were morphometrically evaluated using a semi-quantitative method. A subsample of 76 salmon was additionally evaluated using a quantitative method. Another subsample of 71 salmon was used for reliability testing of the semi-quantitative method. Ten cross-sections of coronary arteries located on the ventral surface of the bulbus arteriosus were semi-quantitatively scored with regard to myointimal coronary lesions. Quantitative measurements comprised maximal intimal thickness and area lesion (%) while semi-quantitative measurements comprised maximal intimal thickness and lesion degree (%). Scores for individual fish were expressed as the average of lesion score, denoted as mean range lesion. One person performed all evaluations blindly. Two pathologists tested repeatability of the semi-quantitative lesion score measurements blindly. Both semi-quantitative and quantitative variables increased significantly with increasing lesion score. Semi-quantitative estimation gave significantly higher maximal intimal thickness values than quantitative measurement. Semi-quantitative measurements were found to explain 63.2% of variation in quantitatively measured maximal intimal thickness. Due to lack of agreement, the two methods are not directly comparable and one method cannot replace the other. Repeatability within and between observers with regard to semi-quantitative classification was very good with Kappa values larger than 61.5%. The semi-quantitative method was a valid and reliable method for coronary lesion evaluation in Atlantic salmon.     

ImproWin? in the treatment of gastric ulceration of the squamous mucosa in trotting racehorses

Background

Gastric ulceration is highly prevalent in horses, and there is a large commercial market for feed-additives and non-licenced products that claim effect for prevention and treatment of gastric ulceration. ImproWin® has been used as a feed additive in horses with anecdotal evidence that it may have some positive effects on gastric ulceration.The aim of this study was to investigate the effect of ImproWin® treatment on spontaneously occurring gastric ulcers of the squamous mucosa in Standardbred and Coldblooded trotting racehorses.The study was performed as a randomised, double-blinded, single centre study with stratified semi cross-over design with breed as stratification factors. The horses were clinically and endoscopically examined prior to start and after three weeks of treatment. The ulcerations were scored in accordance with Equine Gastric Ulcer Council (EGUC) recommendations on a 5 point scale and on a 10 cm Visual Analogue Scale (VAS). The patients were responder-classified after 3 weeks. Responders in need of ulcer treatment were randomly allocated to 2 or 4 weeks of additional treatment. Non-responders to placebo were crossed to ImproWin®.

Results

The 5-point EGUC score and VAS recorded score was significantly reduced (P ≤ 0.01) in both groups after 3 weeks of treatment. From 3 weeks to the end of treatment the score was further significantly reduced in the ImproWin® group (P ≤ 0.05).At the end of treatment, 78% in the ImproWin® group and 54.8% in the placebo group were classified as responders. The difference was significant (P = 0.04).

Conclusions

ImproWin® may aid the healing process of ulcers of the gastric squamous mucosa of trotters.     

The effect of feeding three anabolic steroids in different combinations on the growth,food conversion efficiency and protein and nucleic acid levels of liver,kidney, brain and muscle of mirror carp (Cyprinus carpio)

A study on the effect of three different anabolic-androgenic steroids on the growth, food conversion efficiency and nucleic acid contents of liver, kidney, brain and muscle of carp,Cyprinus carpio was undertaken. The three steroids, methyltestosterone (MT), ethylestrenol (EE) and oxandrolone (ON), were fed in different combinations at final concentrations of 5 or 6 mg/kg diet for 60 days. No effect on the growth was observed in any of the experimental groups. A decrease in the specific growth rate (11–21%) and food conversion efficiency (20–29%) was noted. Feeding of drugs increased the cranio-somatic and reno-somatic index in all except one group. Hepatosomatic (ON+MT) and viscero-somatic (ON+MT; EE+MT+ON) indices decreased. Protein increased and RNA/DNA decreased in only one group while a decrease in protein/DNA was observed in the liver of all experimental groups. RNA/DNA increased and protein/RNA decreased only in one group while no effect was seen in protein and protein/DNA contents in any of the treated kidneys. Proteins, protein/RNA and protein/DNA decreased in certain groups in brain tissue. In muscle, no effect was seen in proteins or protein/DNA. Protein/RNA decreased in all but one group while RNA/DNA was higher only in the group fed all the three steroids together.     

Changes in various metabolic parameters in blood and milk during experimental Escherichia coli mastitis for primiparous Holstein dairy cows during early lactation

Background

The objective of this study was to characterize the changes in various metabolic parameters in blood and milk during IMI challenge with Escherichia coli (E. coli) for dairy cows during early lactation. Thirty, healthy primiparous Holstein cows were infused (h = 0) with ~20-40 cfu of live E. coli into one front mammary quarter at ~4-6 wk in lactation. Daily feed intake and milk yield were recorded. At –12, 0, 3, 6, 12, 18, 24, 36, 48, 60, 72, 96, 108, 120, 132, 144, 156, 168, 180 and 192 h relative to challenge rectal temperatures were recorded and quarter foremilk was collected for analysis of shedding of E. coli. Composite milk samples were collected at -180, -132, -84, -36, -12, 12, 24, 36, 48, 60, 72, 84, 96, 132 and 180 h relative to challenge (h = 0) and analyzed for lactate dehydrogenase (LDH), somatic cell count, fat, protein, lactose, citrate, beta-hydroxybutyrate (BHBA), free glucose (fglu), and glucose-6-phosphate (G6P). Blood was collected at -12, 0, 3, 6, 12, 18, 24, 36, 60, 72, 84, 132 and 180 h relative to challenge and analyzed for plasma non-esterified fatty acids (NEFA), BHBA and glucose concentration. A generalized linear mixed model was used to determine the effect of IMI challenge on metabolic responses of cows during early lactation.

Results

By 12 h, E. coli was recovered from challenged quarters and shedding continued through 72 h. Rectal temperature peaked by 12 h post-challenge and returned to pre-challenge values by 36 h post-IMI challenge. Daily feed intake and milk yield decreased (P <0.05) by 1 and 2 d, respectively, after mastitis challenge. Plasma BHBA decreased (12 h; P <0.05) from 0.96 ± 1.1 at 0 h to 0.57 ± 0.64 mmol/L by 18 h whereas concentration of plasma NEFA (18 h) and glucose (24 h) were significantly greater, 11 and 27%, respectively, after challenge. In milk, fglu, lactose, citrate, fat and protein yield were lower whereas yield of BHBA and G6P were higher after challenge when compared to pre-challenge values.

Conclusions

Changes in metabolites in blood and milk were most likely associated with drops in feed intake and milk yield. However, the early rise in plasma NEFA may also signify enhanced adipose tissue lipolysis. Lower concentrations of plasma BHBA may be attributed to an increase transfer into milk after IMI. Decreases in both milk lactose yield and % after challenge may be partly attributed to reduced conversion of fglu to lactose. Rises in G6P yield and concentration in milk after challenge (24 h) may signify increased conversion of fglu to G6P. Results identify changes in various metabolic parameters in blood and milk after IMI challenge with E. coli in dairy cows that may partly explain the partitioning of nutrients and changes in milk components after IMI for cows during early lactation.     

The cytochrome P450 system of Atlantic salmon (Salmo salar): II. Variations in hepatic catalytic activities and isozyme patterns during an annual reproductive cycle

A group of Atlantic salmon (Salmo salar) was followed through their first year of maturation and spawning. At monthly intervals, starting with juvenile fish in December, 5–7 fish of each sex were killed, and liver and plasma were sampled. The last sampling point was of spawning fish in November a year later. Variables in the cytochrome P450 (P450) system were studied in hepatic microsomes, and estradiol 17 was measured in the plasma of females to assess the maturational status. The P450 1A1-mediated 7-ethoxyresorufin O-deethylase (EROD) started at high levels in winter, but decreased to non-detectable activities in pre-spawning females. Decreases, but not to the same extent, were also observed during this period in total cytochrome P450, cytochrome b₅, NADPH-cytochrome P450 reductase, and in the content of two immunochemically determined P450 isozymes. At the same time, LSI levels increased in maturing females (starting in July), and GSI levels increased in both sexes (starting in May). Sex specific differences were observed in pre-spawning fish in September and October, with levels of total P450, b₅, NADPH-cytochrome P450 reductase, EROD and P450 isozymes significantly lower in females. At the same time, plasma estradiol-17 levels reached peak values in females. The results point to the important role of sex steroids such as estradiol-17 as major factors in the regulation of final sexual maturation. However, this study also indicates that there may be estradiol-17 independent events of equal importance in the early stages of gonadal maturation that may involve the P450 system. The changes observed in the P450 system (as a major drug and steroid metabolizing system) of Atlantic salmon during sexual maturation may be of importance both in the endogenous transduction of hormonal signals, and as a pharmacological basis for designing therapeutic treatment of diseases in the aquaculture industry.Parts of this work were presented at the 5th International Symposium on Responses of Marine Organisms to Pollutants, April 1989 in Plymouth, United Kingdom (Larsen and Goks\/oyr 1989).     

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.     

Macronutrient (N, P, K) and Redoximorphic Metal (Fe, Mn) Allocation in Leersia oryzoides (Rice Cutgrass) Grown Under Different Flood Regimes

Vegetated drainages are an effective method for removal of pollutants associated with agricultural runoff. Leersia oryzoides, a plant common to agricultural ditches, may be particularly effective in remediation; however, research characterizing responses of L. oryzoides to flooding are limited. Soil reduction resulting from flooding can change availability of nutrients to plants via changes in chemical species (e.g., increasing solubility of Fe). Additionally, plant metabolic stresses resulting from reduced soils can decrease nutrient uptake and translocation. The objective of this study was to characterize belowground and aboveground nutrient allocation of L. oryzoides subjected to various soil moisture regimes. Treatments included: a well-watered and well-drained control; a continuously saturated treatment; a 48-h pulse-flood treatment; and a partially flooded treatment in which water level was maintained at 15 cm below the soil surface and flooded to the soil surface for 48 h once a week. Soil redox potential (Eh, mV) was measured periodically over the course of the 8-week experiment. At experiment termination, concentrations of Kjeldahl nitrogen, phosphorus (P), potassium (K), iron (Fe), and manganese (Mn) were measured in plant tissues. All flooded treatments demonstrated moderately reduced soil conditions (Eh < 350 mV). Plant Kjeldahl nitrogen concentrations demonstrated no treatment effect, whereas P and K concentrations decreased in aboveground portions of the plant. Belowground concentrations of P, Mn, and Fe were significantly higher in flooded plants, likely due to the increased solubility of these nutrients resulting from the reductive decomposition of metal–phosphate complexes in the soil and subsequent precipitation in the rhizosphere. These results indicate that wetland plants may indirectly affect P, Mn, and Fe concentrations in surface waters by altering local trends in soil oxidation–reduction chemistry.     

Estimating genetic relationships among historical sources of alfalfa germplasm and selected cultivars with sequence related amplified polymorphisms

Fifteen alfalfa populations consisting of six public cultivars and nine historically recognized sources of alfalfa germplasm in North American cultivars were examined using sequence related amplified polymorphisms (SRAPs). Three bulk DNA samples from each population were evaluated with fourteen different SRAP primer pairs. This resulted in 249 different amplicons, of which over 90% were polymorphic. A dendrogram from the analysis suggests that the public cultivars are quite diverse from all the historical sources of germplasm. The highest mean genetic similarity among the nine original sources of Medicago germplasm was 0.85 between PI 536535 (Peruvian) and 536536 (Indian), while the lowest (0.47) was between PI 560333 (M. falcata) and 536539 (African). The highest mean genetic similarity among the nine original sources of Medicago germplasm and the public alfalfa cultivars was 0.78 between PI 536532 (Ladak) and Vernal, while the lowest (0.59) was between PI 536539 (African) and Oneida. Relationships based on SRAP analysis appear to generally concur with expected relationships based on fall dormancy. This report demonstrates that SRAPs are a promising marker system for detecting polymorphisms in alfalfa.