Identification of cortisol metabolites with LC-MS/MS in plasma, skin mucus, bile and faeces for stress evaluation of farmed Atlantic salmon.

As a stress hormone, cortisol and more recently its metabolites are analysed when assessing fish stress and welfare status, although the exact identity of these metabolites is not clearly defined for the Atlantic salmon. LC-MS/MS techniques, owing to their specificity, sensitivity and ability to simultaneously identify and measure several relevant compounds, can be useful tools for this purpose. Using the guidelines provided by the European Decision no. 657/2002/EC for validation, the LC-MS/MS method presented here, can reliably identify and quantify cortisol and five of its metabolites (5ß-THF, cortisone, 5ß-DHE, 5ß-THE and ß-cortolone) in bile and faeces, and cortisol and cortisone in skin mucus and blood plasma of farmed Atlantic salmon within 15 min. Identified as the most predominant compound in faeces and bile, 5ß-THE is proposed as a candidate stress biomarker when using these matrices. A decision limit (CCα) below 5 ng/mL, a detection capability (CCß) and a limit of detection (LOD) below 10 ng/mL and a limit of quantitation (LOQ) below 30 ng/mL were typically obtained for most of the compounds. The concentrations of these compounds measured in either non-stressed or stressed fish were all above the CCα, CCß, LOD and the LOQ of the method. The latter consequently demonstrated significant difference in cortisol metabolites concentrations between the two groups of fish. The present study further demonstrates that pooling of samples from several individuals could provide reliable results for farmed fish stress evaluation, when sample materials are insufficient in terms of quantity.

Effect of abomasal infusions of a mixture of octadecenoic acids on milk fat synthesis in lactating cows.

Diets causing milk fat depression (MFD) are known to alter ruminal lipid metabolism leading to the formation of specific biohydrogenation intermediates that exert antilipogenic effects. Several isomers of conjugated linoleic acid (CLA), namely trans-10, cis-12 CLA, cis-10, trans-12 CLA, and trans-9, cis-11 CLA, inhibit mammary lipogenesis in the lactating cow, but ruminal outflow of these biohydrogenation intermediates does not account entirely for the reductions in milk fat synthesis during diet-induced MFD. Milk fat trans-10 18:1 concentrations are consistently increased on diets that cause MFD, suggesting a possible role in the regulation of milk fat secretion. Three rumen-fistulated cows in mid lactation were used in a 3 x 3 Latin square to evaluate the effects of a mixture of 18:1 fatty acid methyl esters (FAME) on milk fat synthesis. Experimental treatments consisted of abomasal infusions of ethanol (control), 6 g/d of trans-10, cis-12 CLA (positive control; CLA), or 247 g/d of a mixture of 18:1 FAME containing (% fatty acids) cis-9 (9.45), cis-12 (3.35), trans-10 (37.3), trans-11 (37.4), and trans-12 (2.66) as major isomers (T181 treatment). Administration of the T181 treatment supplied 92.1 g/d of trans-10 18:1. Infusions were conducted over a 5-d period with a 9-d interval between treatments. Treatments had no effect on dry matter intake, milk yield, or milk protein. Relative to the control, abomasal infusion of T181 and trans-10, cis-12 CLA treatments reduced milk fat secretion by 19.5 and 41.5%, respectively. Even though a direct cause and effect on mammary lipogenesis could not be established, comparisons with published data and considerations of the relative abundance of constituent FAME in treatment T181 implicated trans-10 18:1 as the isomer responsible. In conclusion, current data suggest that trans-10 18:1 potentially exerts antilipogenic effects and may contribute to the reduction in milk fat synthesis during diet-induced MFD in the lactating cow.

Intensify production, transform biomass to energy and novel goods and protect soils in Europe-A vision how to mobilize marginal lands.

The rapid increase of the world population constantly demands more food production from agricultural soils. This causes conflicts, since at the same time strong interest arises on novel bio-based products from agriculture, and new perspectives for rural landscapes with their valuable ecosystem services. Agriculture is in transition to fulfill these demands. In many countries, conventional farming, influenced by post-war food requirements, has largely been transformed into integrated and sustainable farming. However, since it is estimated that agricultural production systems will have to produce food for a global population that might amount to 9.1 billion by 2050 and over 10 billion by the end of the century, we will require an even smarter use of the available land, including fallow and derelict sites. One of the biggest challenges is to reverse non-sustainable management and land degradation. Innovative technologies and principles have to be applied to characterize marginal lands, explore options for remediation and re-establish productivity. With view to the heterogeneity of agricultural lands, it is more than logical to apply specific crop management and production practices according to soil conditions. Cross-fertilizing with conservation agriculture, such a novel approach will provide (1) increased resource use efficiency by producing more with less (ensuring food security), (2) improved product quality, (3) ameliorated nutritional status in food and feed products, (4) increased sustainability, (5) product traceability and (6) minimized negative environmental impacts notably on biodiversity and ecological functions. A sustainable strategy for future agriculture should concentrate on production of food and fodder, before utilizing bulk fractions for emerging bio-based products and convert residual stage products to compost, biochar and bioenergy. The present position paper discusses recent developments to indicate how to unlock the potentials of marginal land.

Trans-9, cis-11 conjugated linoleic acid reduces milk fat synthesis in lactating dairy cows.

Under certain dietary situations, rumen biohydrogenation results in the production of unique fatty acids that inhibit milk fat synthesis. The first of these to be identified was trans-10, cis-12 conjugated linoleic acid (CLA), but others are postulated to contribute to diet-induced milk fat depression (MFD). Our objective was to examine the potential role of trans-9, cis-11 CLA in the regulation of milk fat. In a preliminary study, we used gas-liquid and high-performance liquid chromatography techniques to examine milk fat samples from a diet-induced MFD study and found that an increase in trans-9, cis-11 CLA corresponded to the decrease in milk fat yield. We investigated this further using a CLA enrichment of 9, 11 isomers to examine the biological effect of trans-9, cis-11 CLA on milk fat synthesis. Four rumen-fistulated Holstein cows were randomly assigned in a 4 x 4 Latin square experiment involving 5-d treatment periods and abomasal infusion of 1) ethanol (control), 2) a 9, 11 CLA mix (containing 32% trans-9, cis-11, 29% cis-9, trans-11, and 17% trans-9, trans-11), 3) a trans-9, trans-11 CLA supplement, and 4) a trans-10, cis-12 CLA supplement (positive control). The trans-9, trans-11 CLA and trans-10, cis-12 CLA supplements were of high purity (>90%), and all supplements were infused at a rate to provide 5 g/d of the CLA isomer of interest. Milk yield and dry matter intake did not differ among treatments. Compared with the control treatment, milk fat yield was reduced by 15% for the 9, 11 CLA mixture and by 27% for the trans-10, cis-12 CLA treatment. We also found that trans-9, trans-11 CLA had no effect on milk fat yield, and previous research has shown that milk fat yield is unaltered when cows are infused with cis-9, trans-11 CLA. When all treatments were considered, results suggested that trans-9, cis-11 was the CLA isomer in the 9, 11 CLA mix responsible for the reduction in milk fat synthesis, although the magnitude was less than that observed for trans-10, cis-12 CLA. Interestingly, trans-9, trans-11 CLA altered the milk fat desaturase index, further demonstrating that alterations in desaturase can occur independently of effects on milk fat synthesis. Overall, our investigations identified that an increase in milk fat content of trans-9, cis-11 CLA was associated with diet-induced MFD and provided evidence of a role for this isomer in MFD based on the 15% reduction in milk fat yield with abomasal infusion of a CLA enrichment that supplied 5 g/d of trans-9, cis-11 CLA.

Milk fat synthesis is unaffected by abomasal infusion of the conjugated diene 18:3 isomers cis-6,trans-10, cis-12 and cis-6,trans-8,cis-12.

It has been previously established that trans-10,cis-12 CLA is a potent inhibitor of milk fat synthesis. Although the mechanism of this action is not completely understood, it has been speculated that eicosanoid-like metabolites of this isomer formed by the activity of tissue desaturases may be responsible for its activity. The objective of this study was to investigate the effects of an enrichment containing an 18:3 conjugated diene, produced in the metabolism of trans-10,cis-12 CLA, on milk fat synthesis. Three rumen-fistulated Holstein cows (210+/-8 d in milk) were randomly assigned in a 3 x 3 Latin square experiment. Treatments were (i) control, (ii) trans-10,cis-12 CLA supplement (2.1 g/d; positive control), (iii) enrichment providing two conjugated diene 18:3 isomers (2.6 g/d of cis-6,trans-10,cis-12 and 4.0 g/d of cis-6,trans-8,cis-12) and trans-10,cis-12 CLA (2.1 g/d). Treatments were abomasally infused for 5 d at 4-h intervals, and there was a 7-d interval between periods. Milk yield, dry matter intake, and milk protein yield were unaffected by treatments. In contrast, the trans-10,cis-12 CLA supplement reduced milk fat yield by 27%, whereas the supplement enriched with conjugated diene 18:3 isomers (treatment iii) had no effect on milk fat yield beyond that attributable to its trans-10,cis-12 CLA content. The transfer efficiency of trans-10,cis-12 CLA into milk fat was 25 and 24% for treatments ii and iii, respectively. At the same time, the abomasally infused conjugated diene 18:3 isomers were transferred to milk fat with an efficiency of 33 and 41% for cis-6,trans-10,cis-12 and cis-6,trans-8,cis-12 18:3, respectively. Overall, short-term abomasal infusion of the conjugated diene 18:3 isomers had no effect on milk fat synthesis, thereby offering no support for an involvement of metabolites of trans-10,cis-12 CLA in the regulation of milk fat synthesis.

Particulate Matter deposition on Quercus ilex leaves in an industrial city of central Italy.

A number of studies have focused on urban trees to understand their mitigation capacity of air pollution. In this study particulate matter (PM) deposition on Quercus ilex leaves was quantitatively analyzed in four districts of the City of Terni (Italy) for three periods of the year. Fine (between 0.2 and 2.5 µm) and Large (between 2.5 and 10 µm) PM fractions were analyzed. Mean PM deposition value on Quercus ilex leaves was 20.6 µg cm(-2). Variations in PM deposition correlated with distance to main roads and downwind position relatively to industrial area. Epicuticular waxes were measured and related to accumulated PM. For Fine PM deposited in waxes we observed a higher value (40% of total Fine PM) than Large PM (4% of total Large PM). Results from this study allow to increase our understanding about air pollution interactions with urban vegetation and could be hopefully taken into account when guidelines for local urban green management are realized.

Chemical composition of vegetation along urbanisation gradients in two European cities.

Accumulation of particulate matter (PM) and metals on leaves of three deciduous woody species was studied along urbanisation gradients in Stavanger and Warsaw. Differences between rural and urban sites explained most of the observed variation in leaf chemistry, followed by differences between regions. Highest leaf accumulation of elements was found in Warsaw, but also composition of elements differed between the cities. Overall, species showed similar patterns of element accumulation, but differed in accumulation of specific elements. These differences could in part be explained by differences in epicuticular waxes and PM accumulation. Expected source of elements and their chemical characteristics did not explain the observed accumulation patterns. A better differentiation between elements taken up from soil and air would be required for his. Species specific accumulation of elements has to be taken into consideration using leaf samples for biomonitoring.

Yersinia enterocolitica infection--an unrecognized cause of acute and chronic neurological disease? A 10-year follow-up study on 458 hospitalized patients.

The purpose of the present study was to obtain further information about nervous system involvement associated with Yersinia enterocolitica infection. During the period 1974-1983, the infection was diagnosed by antibody response (agglutination titer or ELISA) and/or isolation of the microorganism in 458 hospitalized patients. 2 patients presented with acute symptoms of the peripheral nervous system, and 6 with symptoms of the central nervous system. The 458 patients were followed for 4-14 years (until 1987). During the follow-up period another 6 patients developed chronic neurological conditions; 4 with peripheral symptoms, and 2 with central symptoms. 11 of the 14 patients experienced persistent complaints. In 1991 (10-17 years after the diagnosis of yersiniosis), 6 patients still had significant antibody response (ELISA). The possibility that the immunologically competent Yersinia enterocolitica might have initiated the chronic neurological disease in these patients should not be disregarded.

Could development of malignant mesothelioma be induced by Yersinia enterocolitica infection?

Two out of 458 hospitalized patients with Yersinia enterocolitica infection developed malignant mesothelioma of pleura viz. pericard; both died after a few months. Malignant mesothelioma of the pleura is commonly related to asbestos exposure, whereas pericardiac mesothelioma is an extremely uncommon neoplasm. The possible promotion of malignant mesothelioma by the Yersinia enterocolitica infection should not be disregarded, as the infection may launch chronic immunological reactions resembling those observed among asbestos workers.

Yersinia enterocolitica: an inducer of chronic inflammation.

The aim of the present study was to elucidate the connection between yersiniosis and chronic inflammation. During the period 1974-83, Yersinia enterocolitica infection was diagnosed in 458 hospitalized patients by antibody response, or isolation. The patients were followed for 4-14 years (1987); 160 were readmitted with chronic disease. Fifty-three patients had persistent joint complaints, 18 developed ankylosing spondylitis, 14 rheumatoid arthritis, and 17 iridocyclitis. Thirty-eight patients suffered from chronic abdominal pain, and another 28 from chronic diarrhoea. Two who underwent proctocolectomy microscopically had ulcerative colitis. Eleven patients developed neurological disease; others developed conditions such as chronic nephritis, thyroid disease, insulin-dependent diabetes, etc. Chronic hepatitis, found in 22 patients, was significantly correlated with positive test for antinuclear antibody and rheumatoid factor, and with death. Several patients developed chronic multiorgan disease, probably with chronic hepatitis as pivot. Regarding the whole material, the difference between observed and expected cumulative survival rates remained significant for 8 years (0.9189 < 0.9456; p < 0.025), indicating a substantial impact on long-term survival exerted by chronic yersiniosis.

Contralateral inguinal hernial development and ipsilateral recurrence following unilateral hernial repair in infants and children.

The aim of the present study was to evaluate the frequency of contralateral hernial development and ipsilateral recurrence following surgery for unilateral inguinal hernia in infants and children. During the period 1961-1970, 161 infants and children (143 males and 18 females; 0-12 years old) underwent surgery for unilateral inguinal hernia. The patients were followed for 20-29 years (until 1990). 16/143 males (11.2%), and 1/18 females developed a contralateral inguinal hernia; the frequency among patients less than seven years old was significantly higher than the frequency among older children (16/118 > 1/43; p < 0.05). No significant difference was observed between patients primarily operated for rightsided and leftsided hernias. Four/143 boys (2.7%) experienced testicular maldescens. Recurrence after herniotomy was seen in 9/143 males (6.3%), and in 1/18 females. The low frequency of contralateral hernial development, as the risk of surgical complications, indicate that routine contralateral inguinal exploration may not be recommended while operating upon unilateral symptomatic inguinal hernias. Our recurrence rate of 6.2% may mirror the conditions in a general surgical department, where several surgeons and residents operate upon a limited number of paediatric patients.

Small bowel diverticulosis complicated by diverticulitis and paralytic ileus. Report of a case.

Small bowel diverticulosis occurs in 0.3-4.4% of all autopsies. Generally the condition is clinically silent; but acute diverticulitis with peritonitis and paralytic ileus may occur in elderly people. This complication has a high mortality and its successful management requires early diagnosis and surgery.

Accumulation of particulate matter and trace elements on vegetation as affected by pollution level, rainfall and the passage of time.

Particulate matter is harmful to human health. To reduce its concentration in air, plants could be used as biological filters, accumulating particulate matter on their foliage. In a study carried out at three sites with differing pollution levels and exposure to precipitation, the capacity of evergreen species (Taxus baccata L., Hedera helix L. and Pinus sylvestris L.) to accumulate particulate matter and trace elements from ambient air in urban areas was investigated. The effects of rainfall and the passage of time on particulate matter deposition on foliage were also determined. The results showed that foliage accumulated an increasing quantity of particulate matter in successive months, but the actual amount of particulate matter and trace elements accumulated differed considerably between sites and plant species. The greatest accumulation of air pollutants occurred on the foliage of plants protected from the rain at a site exposed to traffic related pollution and the smallest accumulation at a rural site. Among the species analysed, the deposited mass of particulate matter and trace elements was the greatest on P. sylvestris. In all species, precipitation removed a considerable proportion of particles accumulated on foliage. Most of the removed particulate matter was large size fraction, but little belong to the smallest size fraction. These results showed that both, the dynamics of deposition and leaf washing by rain during the season need to be considered when evaluating the total effect of vegetation in pollutant remediation.

Plant species differences in particulate matter accumulation on leaf surfaces.

Particulate matter (PM) accumulation on leaves of 22 trees and 25 shrubs was examined in test fields in Norway and Poland. Leaf PM in different particle size fractions (PM(10), PM(2.5), PM(0.2)) differed among the species, by 10- to 15-folds at both test sites. Pinus mugo and Pinus sylvestris, Taxus media and Taxus baccata, Stephanandra incisa and Betula pendula were efficient species in capturing PM. Less efficient species were Acer platanoides, Prunus avium and Tilia cordata. Differences among species within the same genus were also observed. Important traits for PM accumulation were leaf properties such as hair and wax cover. The ranking presented in terms of capturing PM can be used to select species for air pollution removal in urban areas. Efficient plant species and planting designs that can shield vulnerable areas in urban settings from polluting traffic etc. can be used to decrease human exposure to anthropogenic pollutants.

Liver affection correlated to low antibody titres against Yersinia enterocolitica. A statistical analysis.

I have previously suggested that Yersinia enterocolitica may cause a chronic liver affection. The aim of the present study has been to provide a statistical support for this hypothesis. During a one-year period, 62 consecutive patients admitted to our department, clinically presenting with conditions often associated with Y.ent. infection, were examined for Y.ent. antibody titre and serum concentrations of bilirubin and the enzymes ASAT, LD and AP. The statistical analysis showed that Y.ent. antibody titres (in range negative to 320) were significantly correlated (p = 0.01) to serum bilirubin concentrations. The microorganism Yersinia enterocolitica should hence be considered as possible aetiological agent in cases of hepatic affection.

The Yersinia enterocolitica infection in acute abdominal surgery. A clinical study with a 5-year follow-up period.

By estimation of antibody titres or possibly isolation of the micro-organism, the role of the Yersinia enterocolitica as an etiological agent in mesenterial lymphadenitis, ileal inflammation, or abdominal colics was evaluated in 94 consecutive patients (25 operated upon and 69 treated conservatively) during a 1-year period from 1975-76. Eighty four patients (24 operated upon and 60 treated conservatively) were re-examined 5 years later. Among the 25 patients operated upon, three presented acute terminal ileitis, two of them in association with acute Y. enterocolitica infection. A substantial part of the 22 patients with other peroperative findings also presented trace or positive Y. enterocolitica antibody titres. Some of these patients experienced complaints attributable to the Y. enterocolitica infection over the following years. At re-examination, previously elevated Y. enterocolitica antibody titres had generally normalized, and in the two patients who in 1975 presented acute terminal ileitis associated with acute Y. enterocolitica infection, radiological examination now demonstrated normal terminal ileum. Among 69 patients treated conservatively for abdominal colics, 36 in 1975 presented trace or positive titres. During the 5-year follow-up period, abdominal pain and joint complaints were significantly more commonly experienced by patients with trace or positive titres than by patients with negative titres (p = 0.007 and p = 0.004, respectively). The Y. enterocolitica is an important cause of abdominal disease in our region, and the infection commonly gives symptoms of long duration.

Pneumoperitoneum associated with perforated appendicitis.

Perforated appendicitis is sometimes accompanied by pneumoperitoneum, often localized beneath the diaphragmatic dome. This may lead to very dangerous diagnstoic errors, which every roentgenologist and surgeon should be aware of. A historical review is followed by three case reports.

Liver affection associated with Yersinia enterocolitica infection.

During the last ten years, several clinical manifestations of Yersinia enterocolitica infection have been reported. Surgeons are especially aware of "the right iliac fossa syndrome", caused by mesenterial lymphadenitis and terminal ileitis. We suggest that Yersinia enterocolitica may also cause a clinical condition easily misinterpreted as cholecystitis, and accompanied by slightly elevated serum levels of ASAT, LD, AP and bilirubin. Apparently, this condition may run a chronic relapsing course. A report is given of two cases of liver affection associated wtih positive Y. ent. antibody titre. Case 1 would illustrate the chronic relapsing liver affection with stationary titre. In Case 2 an acute Au-negative hepatitis is accompanied by significant rise and fall in titre.

A survey of acute and chronic disease associated with Yersinia enterocolitica infection. A Norwegian 10-year follow-up study on 458 hospitalized patients.

The aim of the present study was to further elucidate acute and chronic manifestations of Yersinia enterocolitica infection. During the period 1974-83, 458 hospitalized patients were diagnosed by antibody response and/or isolation of the microorganism. 64 patients had suffered from chronic conditions as rheumatic disease, inflammatory bowel disease, hepatitis, nephritis or thyroid disease for some time. Acute hepatic, renal, cardiac, pulmonary, pancreatic or neurologic involvement were observed in a substantial portion of patients; several had multiorgan disease. Acute insulin-dependent diabetes was seen in 2 patients, malignant mesothelioma in 2, and specific lymph node inflammation in 1. The patients were followed for 4-14 years (1987). 36/160 readmitted patients had abdominal pain and 26 had diarrhea; chronic colitis was demonstrated in 4. Some patients developed rheumatic conditions; others developed chronic disease of liver, kidneys, heart, pancreas, thyroid or nervous system. Chronic liver disease, in 22 patients, was correlated with positive tests for antinuclear antibody and rheumatoid factor; and might influence development of malignant disease, and mortality. A variety of acute and chronic clinical pictures may be associated with Y. enterocolitica infection, and further clinical research is required in this field.