2-Amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP) level in human hair as biomarkers for dietary grilled/stir-fried meat and fish intake

Several case-control studies have reported possible associations between heterocyclic amine (HCA) intake and the risk of cancer. However, the validity of a questionnaire to assess HCA intake has hardly been examined. In particular, no biomarker which could serve as an independent measure of habitual HCA intake has been established. Therefore, the validity of a questionnaire to assess HCA intake by means of a biomarker remains to be investigated. In this study, we examined the availability of hair HCAs as a biochemical indicator of dietary intake of HCAs. Study subjects were 20 volunteers (7 men and 13 women) aged 25–57 years, either residents of Tokyo or the neighboring cities in Japan. We collected individual weighed dietary records (DR) over 28 consecutive days. Approximately 3–5 g of hair was collected twice from all subjects before and after DR at intervals of 1–3 months. The mean (S.D.) 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP) level of hair was 1376.0 pg/g hair (928.9) and 16.6 ng/g melanin (12.3). A steady increase in the mean PhIP level in hair from the lowest to the highest tertile of the grilled/stir-fried meat intake was observed (P = 0.009), but not in the grilled/stir-fried fish intake (P = 0.461). The PhIP level in hair was highly correlated with the grilled/stir-fried meat intake (r = 0.68) but not with the grilled/stir-fried fish intake (r = 0.28). These observations were made of hair with and without melanin adjustment. The present study indicates that the PhIP level in hair can be used as a biological indicator of dietary intake of HCAs.     

Validity of a self-administered food frequency questionnaire in the assessment of heterocyclic amine intake using 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP) levels in hair

Several case-control studies have reported possible associations between heterocyclic amine (HCA) intake and the risk of cancer. The validity of questionnaires used to assess HCA intake has hardly been examined, however; in particular, no biomarker able to serve as an independent measure of habitual HCA intake has been established. In this study, we examined the validity of HCA intake estimated from a food frequency questionnaire (FFQ) using 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP) level in hair as a reference method. Study subjects were 20 volunteers (7 men and 13 women) aged 25-57 years residing in Tokyo or neighboring cities in Japan. The subjects completed the FFQ, and gave 3-5g of hair twice at an interval of 1-3 months for use in establishing validity. Results showed that intakes of PhIP, MeIQ, Trp-P-1, and total HCA by the FFQ were significantly correlated with PhIP levels in hair when adjustment was made for melanin content (r=0.47, r=0.50, r=0.55, and r=0.51, respectively). The present study indicates that HCA intake estimated from this FFQ provides a reasonable ranking of individuals to allow the analysis of associations between HCA intake and risk of cancer in large-scale epidemiological studies.     

2-Amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP) level in human hair as biomarkers for dietary grilled/stir-fried meat and fish intake

Several case-control studies have reported possible associations between heterocyclic amine (HCA) intake and the risk of cancer. However, the validity of a questionnaire to assess HCA intake has hardly been examined. In particular, no biomarker which could serve as an independent measure of habitual HCA intake has been established. Therefore, the validity of a questionnaire to assess HCA intake by means of a biomarker remains to be investigated. In this study, we examined the availability of hair HCAs as a biochemical indicator of dietary intake of HCAs. Study subjects were 20 volunteers (7 men and 13 women) aged 25-57 years, either residents of Tokyo or the neighboring cities in Japan. We collected individual weighed dietary records (DR) over 28 consecutive days. Approximately 3-5 g of hair was collected twice from all subjects before and after DR at intervals of 1-3 months. The mean (S.D.) 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP) level of hair was 1376.0 pg/g hair (928.9) and 16.6 ng/g melanin (12.3). A steady increase in the mean PhIP level in hair from the lowest to the highest tertile of the grilled/stir-fried meat intake was observed (P = 0.009), but not in the grilled/stir-fried fish intake (P = 0.461). The PhIP level in hair was highly correlated with the grilled/stir-fried meat intake (r = 0.68) but not with the grilled/stir-fried fish intake (r = 0.28). These observations were made of hair with and without melanin adjustment. The present study indicates that the PhIP level in hair can be used as a biological indicator of dietary intake of HCAs.     

Associations of maternal prenatal dietary intake of n-3 and n-6 fatty acids with maternal and umbilical cord blood levels

Maternal n-3 and n-6 polyunsaturated fatty acid (PUFA) status may influence birth outcomes and child health. We assessed second trimester maternal diet with food frequency questionnaires (FFQs) (n=1666), mid-pregnancy maternal erythrocyte PUFA concentrations (n=1550), and umbilical cord plasma PUFA concentrations (n=449). Mean (SD) maternal intake of total n-3 PUFA was 1.17 g/d (0.43), docosahexaenoic and eicosapentaenoic acids (DHA+EPA) 0.16 g/d (0.17), and total n-6 PUFA 12.25 g/d (3.25). Mean maternal erythrocyte and cord plasma PUFA concentrations were 7.0% and 5.2% (total n-3), 5.0% and 4.6% (DHA+EPA), and 27.9% and 31.4% (total n-6). Mid-pregnancy diet–blood and blood–blood correlations were strongest for DHA+EPA (r=0.38 for diet with maternal blood, r=0.34 for diet with cord blood, r=0.36 for maternal blood with cord blood), and less strong for n-6 PUFA. The FFQ is a reliable measure of elongated PUFA intake, although inter-individual variation is present     

Serum selenium and selenoprotein P status in adult Danes – 8-year followup

Selenium is an essential micronutrient important to human health. The main objective of this study is to describe serum selenium and selenoprotein P status in two samples of the Danish population. In addition, the influence of various factors potentially associated with selenium status was investigated.Blood samples from a total of 817 randomly selected subjects from two cities in Denmark were analyzed. Half of the samples were collected in 1997–1998 and the other half in 2004–2005. Samples from women aged 18–22, 40–45 and 60–65 years, and men aged 60–65 years were selected for this study. All subjects had filled in a food frequency questionnaire (FFQ) and a questionnaire with information about smoking habits, alcohol consumption and exercise habits.Mean serum selenium level was 98.7±19.8 μg/L and median selenoprotein P level was 2.72 (2.18–3.49) mg/L. Serum selenium and selenoprotein P increased with age, and selenoprotein P was higher in men than in women. Serum selenium levels decreased by 5% on average from 1997–98 to 2004–05 (P<0.001), whereas selenoprotein P level increased (P<0.001). The intake of fish correlated weakly with serum selenium level (r=0.14, P<0.001) but not with selenoprotein P level. Smoking status, alcohol intake, exercise habits, BMI and medicine use did not influence selenium status.It is concluded that selenium status in this Danish population is at an acceptable level. No major groups with regard to age, sex or lifestyle factors could be identified as being in risk for selenium deficiency.     

Corpuscular oxidative stress in desert sheep naturally deficient in copper

Oxidative stress arises when there is an imbalance between radical-generating and radical-scavenging activity; it may therefore cause an increase in oxidation products and cell damage. This study aimed to determine antioxidant status, lipid peroxidation, and their relation to anemia of grazing sheep deficient in copper (Cu). For this purpose, 39 male lambs of native (Balady) breed, aged 6–7 months and reared in El-Dakhla oasis (in the western Egyptian desert), were divided according to plasma Cu (pCu) concentration into three groups, marginally deficient (MD, pCu = 4–8 μmol/l, n = 12), functionally deficient (FD, pCu < 3 μmol/l, n = 12) and control (pCu > 9 μmol/l, n = 15). Jugular blood was sampled for determination of red blood cell count (RBC), packed cell volume (PCV), hemoglobin concentration (Hb), plasma ceruloplasmin activity (pCp), antioxidant activities of erythrocytic superoxide dismutase (eSOD), catalase (eCAT), glutathione peroxidase (eGSH-Px), and levels of erythrocytic malondialdehyde (eMDA, as a biomarker of lipid peroxidation). The Cu-deficient lambs were characterized by microcytic hypochronic anemia accompanied by decreased pCp, eSOD, eCAT and eGSH-Px activities and increased eMDA level when compared to the controls. The indices of anemia, pCp and eSOD were lower and eMDA was higher in FD compared to MD lambs. The enhanced eMDA was strongly correlated (P < 0.01) with the inhibited activity of pCu (r = −0.79), pCp (r = −0.65) and eSOD (r = −0.71) and to a lesser extent (P < 0.05) with eGSH-Px (r = −0.38) and eCAT (r = −0.41). In addition, eMDA was negatively correlated (P < 0.01) with RBC (r = −0.75), PCV (r = −0.69) and Hb (r = −0.72). This study suggests that Cu-deficient lambs incur an erythrocytic oxidative damage secondary to impaired oxidant defenses, which may be one of the mechanisms underlying Cu deficiency-induced anemia in grazing sheep.     

Measurement of body surface area in children with liver disease by a novel three-dimensional body scanning device

Body surface area (BSA) is used in paediatrics to assess fluid requirement, drug doses, cardiac output and glomerular filtration rate. The aim of this study was to examine, in children with liver disease, the relationship between BSA determined by a traditional nomogram and BSA measured by a novel three-dimensional technique — Loughborough Anthropometric Shadow Scanner (LASS). Subjects were 16 children, mean age 8.1 (range 3.6–14.9) years, with a variety of liver diseases. Twenty-eight controls had a mean age of 7.1 (3.1–10.5) years. All had LASS scans performed as well as 21 anthropometric measurements taken by a single observer. There was a significant relationship between BSA (LASS) and BSA nomogram for liver-diseased children (r=0.99) and controls (r=0.96). The BSA nomogram values were significantly greater (P < 0.05) than BSA (LASS) for liver-diseased subjects by 10.1% (–0.35 to + 20.6; 95% confidence interval), and for controls by 9.6% (4.1–23.2). Best prediction of BSA (LASS) for liver-disease subjects used height, body weight and gluteal furrow circumference [r ²=0.997; standard estimated error (SEE) = 0.015 m²] and for controls used body weight alone (r ²=0.907; SEE=0.048 m²). BSA nomogram has no additional error in children with liver disease, but may overestimate BSA by 10% compared with a novel three-dimensional body surface scanning technique.     

The relationship between micronuclei in human lymphocytes and selected micronutrients in vegetarians and non-vegetarians

A vegetarian diet results in higher intake of vitamins and micronutrients, which – although providing antioxidant defence – may lead to deficiency in other micronutrients involved in DNA metabolism and stability (such as vitamins belonging to the B group). The principal difference among various vegetarian diets is the extent to which animal products are avoided. We have performed a pilot study to determine the relationship between the micronucleus frequency in lymphocytes and diet, and we compared the levels of Vitamins C and E, β-carotene, B₁₂, folic acid, homocysteine and total antioxidant capacity in healthy vegetarians and non-vegetarians. The vegetarian group, consisting of 24 volunteers (13 women and 11 men), were matched for age and sex with 24 volunteers (12 women and 12 men) with a traditional dietary habit. Among the vegetarians were 13 lacto-ovo-vegetarians with average duration of vegetarian diet 10.8 years (ranging from 5 to 26 years) and 11 lacto-vegetarians with average duration of vegetarian diet 8.2 years (ranging from 3 to 15 years). Homocysteine, Vitamins C and E and β-carotene levels in plasma were assayed by HPLC, and serum folate and Vitamin B₁₂ were determined with Elecsys Immunoassay tests. The total antioxidant capacity of plasma was estimated by measuring the ferric-reducing activity in a spectrophotometric assay. Micronuclei were measured in cytokinesis-blocked lymphocytes. Vegetarians had significantly higher levels of Vitamin C and β-carotene (but not Vitamin E) in plasma compared with non-vegetarians (P < 0.001). There were no significant differences in serum levels of folic acid and Vitamin B₁₂ between the monitored groups. Levels of folic acid in vegetarians correlated with length of vegetarianism (r = 0.62, P = 0.001, N = 24). Vegetarians had elevated levels of homocysteine compared with non-vegetarians (P = 0.007), as did vegetarian women compared with non-vegetarian women (P = 0.031). We did not find any differences in total antioxidant capacity or in micronucleus frequency between the groups. Micronuclei correlated with age (r = 0.62, P < 0.001, N = 48), women having higher frequencies than men. Multifactorial regression analysis showed significant effects of age, sex and total antioxidant capacity on micronucleus frequency (N = 48, P < 0.001).     

Determinants of anti-benzo[a]pyrene diol epoxide–DNA adduct formation in lymphomonocytes of the general population

We evaluated determinants of anti-benzo[a]pyrenediolepoxide-(B[a]PDE)–DNA adduct formation (adduct induced by the ultimate carcinogenic metabolite of B[a]P) in lymphomonocytes of subjects environmentally exposed to low doses of polycyclic aromatic hydrocarbons (PAHs) (B[a]P). Our study population consisted of 585 Caucasian subjects, all municipal workers living in North-East Italy and recruited during their periodic check-ups after informed consent. PAH (B[a]P) exposure was assessed by questionnaire. Anti-B[a]PDE–DNA levels were measured by HPLC fluorescence analysis.We found that cigarette smoking (smokers (22%) versus non-smokers, p < 0.0001), dietary intake of PAH-rich meals (≥52 (38%) versus <52 times/year, p < 0.0001), and outdoor exposure (≥4 (19%) versus <4 h/day; p = 0.0115) significantly influenced adduct levels. Indoor exposure significantly increased the frequency of positive subjects (≥0.5 adducts/10⁸ nucleotides; χ² for linear trend, p = 0.051). In linear multiple regression analysis the major determinants of increased DNA adduct levels (ln values) were smoking (t = 6.362, p < 0.0001) and diet (t = 4.035, p < 0.0001). In this statistical analysis, indoor and outdoor exposure like other factors of PAH exposure had no influence. In non-smokers, the influence of diet (p < 0.0001) and high indoor exposure (p = 0.016) on anti-B[a]PDE–DNA adduct formation became more evident, but not that of outdoor exposure, as was confirmed by linear multiple regression analysis (diet, t = 3.997, p < 0.0001 and high indoor exposure, t = 2.522, p = 0.012).This study indicates that anti-B[a]PDE–DNA adducts can be detected in the general population and are modulated by PAH (B[a]P) exposure not only with smoking – information already known from studies with limited number of subjects – but also with dietary habits and high indoor exposure. In non-smokers, these two factors are the principal determinants of DNA adduct formation. The information provided here seems to be important, since DNA adduct formation in surrogate tissue is an index of genotoxic exposure also in target organs (e.g., lung) and their increase may also be predictive of higher risk for PAH-related cancers.     

Blood reflects tissue oxidative stress depending on biomarker and tissue studied

This study investigated whether selected oxidative stress markers measured in blood adequately reflect redox status in skeletal muscle, heart, and liver. Several markers were determined after implementing two treatments known to affect redox status, namely exercise and allopurinol administration. Xanthine oxidase, thiobarbituric acid-reactive substances (TBARS), protein carbonyls (PC), reduced glutathione (GSH), oxidized glutathione (GSSG), catalase, and total antioxidant capacity were determined in blood, skeletal muscle, heart, and liver. Correlation between blood and tissues in each marker was performed through the Spearman rank correlation coefficient. GSSG in erythrocytes was correlated with all tissues, ranging in the five experimental groups as follows: skeletal muscle r_s = 0.656–0.874, heart r_s = 0.742–0.981, liver r_s = 0.646–0.855. Xanthine oxidase and TBARS measured in blood satisfactorily described the redox status of the heart (0.753–0.964 and 0.705–1.000, respectively) and liver (0.755–0.902 and 0.656–1.000, respectively). Skeletal muscle and heart redox status can be adequately described by PC (0.652–1.000 and 0.656–0.964, respectively), GSH (0.693–1.000 and 0.656–1.000, respectively), and catalase (0.745–1.000 and 0.656–1.000, respectively) measured in blood. In conclusion, this study suggests that a combination of markers measured in blood provides a reliable indication about the redox status in skeletal muscle, heart, and liver.     

Decomposition and nutrient release by green manures in a tropical hillside agroecosystem

The decomposition and nutrient release of 12 plant materials were assessed in a 20-week litterbag field study in hillsides from Cauca, Colombia. Leaves of Tithonia diversifolia (TTH) and Indigofera constricta (IND) decomposed quickly (k=0.035±0.002 d^–1), while those of Cratylia argentea (CRA) and the stems evaluated decomposed slowly (k=0.007±0.002 d^–1). Potassium presented the highest release rates (k>0.085 d^–1). Rates of N and P release were high for all leaf materials evaluated (k>0.028 d^–1) with the exception of CRA (N and P), TTH and IND (P). While Mg release rates ranged from 0.013 to 0.122 d^–1, Ca release was generally slower (k=0.008–0.041 d^–1). Initial quality parameters that best correlated with decomposition (P>0.001) were neutral detergent fibre, NDF (r=–0.96) and in vitro dry matter digestibility, IVDMD (r=0.87). It is argued that NDF or IVDMD could be useful lab-based tests during screening of plant materials as green manures. Significant correlations (P>0.05) were also found for initial quality parameters and nutrient release, being most important the lignin/N ratio (r=–0.71) and (lignin+polyphenol)/N ratios (r=–0.70) for N release, the C/N (r=0.70) and N/P ratios (r=–0.66) for P release, the hemicellulose content (r=–0.75) for K release, the Ca content (r=0.82) for Ca release, and the C/P ratio (r=0.65) for Mg release. After 20 weeks, the leaves of Mucuna deerengianum released the highest amounts of N and P (144.5 and 11.4 kg ha^–1, respectively), while TTH released the highest amounts of K, Ca and Mg (129.3, 112.6 and 25.9 kg ha^–1, respectively). These results show the potential of some plant materials studied as sources of nutrients in tropical hillside agroecosystems.     

Advantages of the Use of Deciduous Teeth,Hair, and Blood Analysis for Lead and Cadmium Bio-Monitoring in Children. A Study of 6-Year-Old Children from Krakow (Poland)

The population of about 300 6-year-old preschool children was studied for lead (Pb) and cadmium (Cd) in deciduous teeth, scalp hair, and capillary blood. Zinc (Zn) content in hair was also measured due to its possible interferences with the elements studied. The Pb and Cd contents were measured by graphite furnace atomic absorption spectrometry with Zeeman background correction, and Zn, by flame method. Metal levels found were comparable to those in European countries. Positive correlations between Pb and Cd in the same and different tissues were widespread (r = 0.23–0.68). A positive correlation was found between Pb in teeth and blood (r = 0.65, p < 0.001). Pb blood threshold 100 μg L⁻¹ referred to 2.6 μg g⁻¹ Pb in teeth. Predictability of this Pb teeth limit was 14% vs. 5% for Pb blood. Strong negative correlation was found between hair Pb and Zn (r = −0.68, p < 0.001). Enhanced Cd and Zn levels in hair were associated with the household drinking water hardness. The associations with some other environmental and behavioral factors were also presented and discussed. The results proved the advantage of analysis of primary teeth for Pb exposure screening of preschool children.     

Factors regulating circulating vascular endothelial growth factor (VEGF): Association with bone mineral density (BMD) in post-menopausal osteoporosis

Vascular endothelial growth factor (VEGF) plays an important role in bone health. We investigated the factors which influence circulating VEGF and their association with bone mineral density (BMD). Two hundred and fifty two post-menopausal women aged 64.5 [9.2] years were studied. BMD was determined at the lumbar spine (LS), femoral neck (FN) and total hip (TH). Serum oestradiol and VEGF were measured. Subjects were genotyped for two polymorphic variants in the 5′ untranslated region of the VEGF gene; G(634)C and C(936)T. Positive correlations were seen between circulating VEGF and BMI (r = 0.2, p < 0.02) and oestradiol (r = 0.25, p < 0.001). Following multi-linear regression analysis, serum VEGF was associated with the G(634) polymorphism (p = 0.08) and dietary calcium intake (p = 0.02). The association with calcium intake may be mediated by PTH as suggested by the in vitro studies. Following correction for confounders, there was no association between circulating VEGF and BMD at any site. Both VEGF polymorphisms were significant predictors of LS BMD G(634)C: p = 0.017 and C(936)T: p = 0.05. Circulating VEGF may be influenced by genetic, environmental and endocrine factors. Polymorphic variants in the VEGF gene are associated with spine BMD. Further larger studies are needed.     

A radioenzymatic assay of catechol-O-methyltransferase in hair root cells: Comparison with erythrocyte activity

Summary A sensitive radioenzymatic assay of catechol-O-methyltransferase (COMT) in hair root cells is presented. Only five hair roots with intact bulb and sheath are needed for one assay. By pulling 15–20 hairs, 3–4 parallel assays can be performed. As in erythrocytes the COMT activity in hair root cells is constant for each individual. Nevertheless, there is no high correlation between the enzyme activities in erythrocyte and in hair root cells (r=0.26, 0.1> P>0.05, N=46).The determination of COMT in hair root cells offers a further application of this source in genetic research, as in the study of a correlation between COMT activity and various endogenous psychiatric disorders.Part of the thesis of T. Strohmeyer, Faculty of Medicine, University of HamburgDedicated to Prof. H. Holzer on the occasion of his 60th birthday     

Effects of sodium gluconate and phytase on performance and bone characteristics in broiler chickers

An experiment was conducted to evaluate the effects of corn soybean meal diets with added sodium gluconate (0 and 20 g/kg) and phytase (0, 500 and 1000 U/kg) on performance and bone characteristics of broiler chicks. A total of 350 eight-day-old Arbor Acre male chicks were used with a 2 × 3 plus 1 factorial arrangement. A positive control diet, adequate in non-phytate-phosphorus and calcium without sodium gluconate and phytase, was used. Chickens were randomly allocated to seven treatments with each treatment having five replicates. The experiment lasted from 8 to 42 d posthatching. The outcomes of the study indicated that the low-non-phytate-phosphorus diet caused a negative effect (P<0.05) on the average daily gain, average daily feed intake, tibia weight and tibia ash of birds compared to the positive control diet. The supplementation of 20 g sodium gluconate/kg increased average daily gain during days 22–42 (P=0.005), 8–42 (P=0.013) and tibia ash at 21 d (P=0.002). Phytase addition improved (P<0.05) average daily gain and average daily feed intake during the whole experiment and tibia weight, tibia ash at 21 and 42 d and calcium content in tibia ash at 42 d. Compared with diets supplemented with 500 U phytase/kg, diets supplemented with 1000 U phytase/kg had significantly higher average daily feed intake during the whole experiment and average daily gain during days 22–42 and 8–42 and tibia ash at days 21 and 42. There was a significant interaction between sodium gluconate and phytase for average daily gain (P=0.027) from 8 to 21 d and tibia weight (P=0.020) at 42 d. These results demonstrated that sodium gluconate and phytase supplementation to low-phosphorus diets improved performance and phytate-phosphorus utilization by chicks during the whole growing periods.     

A note on alarm cues in juvenile African catfish, Clarias gariepinus Burchell: Indications for opposing behavioural strategies

Following exposure of African catfish to alarm cues, a relationship (r_p = 0.60, N = 10, P = 0.07) between feed efficiency (measured as residual feed intake) and the change in percentage of time spent swimming in response to damage-released alarm cues was observed. Feed-efficient animals responded with a decrease in percentage of time spent swimming (P < 0.05) whereas feed-inefficient animals responded with an increase in percentage of time spent swimming (P < 0.05). This study shows opposing behavioural strategies in African catfish in response to damage-released alarm cues being related to feeding efficiency.     

Seasonal influence on the reproductive performance of swine in the humid zone of Ghana

Relationships between month and climatic factors (ambient temperature, relative humidity, RH, and rainfall) with litter size, conception rate, days from weaning to conception and mortalities at birth and preweaning were studied in large white pigs under research station (RS) and commercial farm (CF) conditions. In RS and CF 868 and 572 farrowing records, respectively, were involved. Litter size was almost evenly distributed in all months of the year, at both farms, despite significant (P<0.05) negative but low correlations between litter size and maximum (r=–0.272, RS) and minimum (r=–0.233, CR) temperatures in the month after conception. There were no significant differences in conception rate between months. However under RS conditions there were significant (P<0.05) correlations between minimum temperature and conception rate in the months prior to (r=–0.362) and at conception (r=–0.221). Days from weaning to conception were bimodally distributed with peak values occurring between January and June and October and December. Significant mortality at birth (P<0.05) and preweaning was generally highest in the cold rainy months of May to October. However, only under CF conditions were there significant correlations between minimum temperature in the month of birth and percentage mortalities at birth (r=–0.217) and preweaning (r=–0.250). Rainfall and RH had no significant (P>0.05) correlation with birth and preweaning mortalities.     

Leaf surface traits: overlooked determinants of birch resistance to herbivores and foliar micro-fungi?

Previous studies on the mechanisms of birch resistance to herbivores and foliar micro-fungi (both pathogenic and endophytic) have focused mainly on the role of internal leaf chemistry. In the present study, we examined genetic correlations between leaf surface traits (glandular trichome density and total concentrations of surface flavonoid aglycones) and occurrence of three species of foliar micro-fungi, one pathogenic rust (Melampsoridium betulinum) and two endophytic fungi (Fusicladium sp. and Melanconium sp.), and performance of autumnal moth larvae (Epirrita autumnata) in two birch species, Betula pubescens ssp. czerepanovii and B. pendula. The performance of autumnal moth larvae on B. pubescens ssp. czerepanovii was negatively correlated with density of glandular trichomes (RGR: r=–0.855; pupal mass: r=–0.709). In addition, rust infection was negatively correlated with trichome density in B. pendula (r=–0.675) and with epicuticular flavonoid aglycones in B. pubescens ssp. czerepanovii (r=–0.855). The frequency of the endophytic fungus Fusicladium sp., was related to epicuticular flavonoid aglycones (r=–0.782), while another endophytic fungus, Melanconium sp., showed no associations with any of the studied variables in B. pubescens ssp. czerepanovii. Our results indicate that leaf surface traits may be at least as important determinants of herbivore performance and micro-fungi abundance in birch as leaf internal chemistry.     

Root health evaluation of three alfalfa varieties in Kerquin sandy land

Without a robust and healthy root system, establishment, productivity, and persistence are compromised. Consequently, research on alfalfa root morphology and health is very important in development of technology for efficient improvement and production of alfalfa. The objectives of this study were to evaluate the root morphology and health of three alfalfa varieties, Algonquin, Golden Queen, and Yellow Flower and to determine relationships among root morphology traits and root health. Yields from these varieties ranged from 5.83 to 43.93 t/ha, total root length ranged from 215.17 to 708.89 mm, root surface area from 124.95 to 468.37 cm², volume from 3.24 to 57.72 cm³, and forks from 1.25 × 10³ to 10.54 × 10³, and tips from 0.65 × 10³ to 3.17 × 10³. Root infestation score was negatively correlated with yield (r = ?0.997, P < 0.01), and was positively correlated with all root morphology traits (r = 0.466–0.997, P < 0.01), and yield was negatively associated with root morphology traits (r = ?0.755 to ?0.998, p < 0.01) with the exception of root tips (r = 0.448, P < 0.01). Results from these analyses indicated that root infestation score was the lowest averaged over age of alfalfa stand in Algonquin. Yield in 2-year old stands was greater in Golden Queen compared to the other two cultivars.     

The chlorophenoxy herbicide dicamba and its commercial formulation banvel induce genotoxicity and cytotoxicity in Chinese hamster ovary (CHO) cells

The sister chromatid exchange (SCE) frequency, the cell-cycle progression analysis, and the single cell gel electrophoresis technique (SCGE, comet assay) were employed as genetic end-points to investigate the geno- and citotoxicity exerted by dicamba and one of its commercial formulation banvel^® (dicamba 57.71%) on Chinese hamster ovary (CHO) cells. Log-phase cells were treated with 1.0–500.0 μg/ml of the herbicides and harvested 24 h later for SCE and cell-cycle progression analyses. All concentrations assessed of both test compounds induced higher SCE frequencies over control values. SCEs increased in a non-dose-dependent manner neither for the pure compound (r = 0.48; P > 0.05) nor for the commercial formulation (r = 0.58, P > 0.05). For the 200.0 μg/ml and 500.0 μg/ml dicamba doses and the 500.0 μg/ml banvel^® dose, a significant delay in the cell-cycle progression was found. A regression test showed that the proliferation rate index decreased as a function of either the concentration of dicamba (r = −0.98, P < 0.05) or banvel^® (r = −0.88, P < 0.01) titrated into cultures in the 1.0–500.0 μg/ml dose-range. SCGE performed on CHO cells after a 90 min pulse-treatment of dicamba and banvel^® within a 50.0–500.0 μg/ml dose-range revealed a clear increase in dicamba-induced DNA damage as an enhancement of the proportion of slightly damaged and damaged cells for all concentrations used (P < 0.01); concomitantly, a decrease of undamaged cells was found over control values (P < 0.01). In banvel^®-treated cells, a similar overall result was registered. Dicamba induced a significant increase both in comet length and width over control values (P < 0.01) regardless of its concentration whereas banvel^® induced the same effect only within 100.0–500.0 μg/ml dose range (P < 0.01). As detected by three highly sensitive bioassays, the present results clearly showed the capability of dicamba and banvel^® to induce DNA and cellular damage on CHO cells.