Induced ovulation and embryonic development of ocellated puffer, Takifugu ocellatus

Acute injections of different hormones to induce ovulation in mature ocellated puffer, Takifugu ocellatus, collected from natural waters during the spawning season, were carried out to develop a reliable protocol for mass production of seed in this species. All experimental fish were divided into seven groups treated with: a saline injection (control), single or two injections of luteinizing hormone‐releasing hormone analog (LHRH‐a; single injection: 50 μg kg^?1, two injections: 10 and 40 μg kg^?1), single or two injections of pituitary (single injection: 6 mg kg^?1, two injections: 1 and 5 mg kg^?1) and single or two injections of human chorionic gonadotropin (hCG; single injection: 2500 IU kg^?1, two injections: 500 and 2000 IU kg^?1), respectively. The percentage of fish that ovulated in six hormonal treatments reached 100%, either with a single injection or with two injections whereas the fish in control group failed to spawn. There were no significant differences among all hormonal treatments in egg production, fertilization rate, or hatch rate (P > 0.05) except time to ovulation between a single injection group and the two‐injection group (P < 0.05). The fertilized eggs of ocellated puffer were spherical, demersal, and adhesive. They had a mean oocyte diameter of 1.487 ± 0.106 mm (range: 1.404–1.560). The egg membrane was transparent and yolk was buff in color, containing a cluster of small oil globules. Thirty‐four successive stages of embryonic development were identified and characterized. Fertilized eggs incubated at 18–20°C generally commenced hatching at 144 h after fertilization. Newly hatched larvae were about 3.26–3.45 mm in length. The induced ovulation technique using acute injections of hormones is an important step in the development of the culture of the ocellated puffer.     

Feed intake and tissue distribution of florfenicol in cod (Gadus morhua) administered feed with different fat contents

This study was performed to examine the appetite and the corresponding plasma and tissue distribution of florfenicol when administered to healthy groups of cod using medicated and non‐medicated salmonid and marine feeds. Marine feed contains approximately 18% fat whereas salmonid feed contains approx. 30% fat. Two groups of fish were medicated with florfenicol at a dosage of 10 mg kg^?1 day^?1 for 10 consecutive days when the drug was administered either via marine or salmonid pellets. Two groups of fish also received either non‐medicated marine or salmonid pellets. Twenty‐four hours after giving the medicated marine feed, 14 out of 20 fish contained detectable concentrations of florfenicol with mean values (n = 14) of 4.67 ± 4.02 μg ml^?1 in plasma, 2.29 ± 2.11 μg g^?1 in muscle and 0.79 ± 0.69 μg g^?1 in the liver. In the fish given medicated salmonid feed, 18 of 20 fish contained detectable concentrations of florfenicol with mean values (n = 18) of 1.77 ± 1.84 μg ml^?1 in plasma, 0.75 ± 0.66 μg g^?1 in muscle and 0.30 ± 0.25 μg g^?1 in the liver. Decreased feed intake of the salmonid feed, both medicated and non‐medicated, was noted when compared to medicated and non‐medicated marine feed. No difference in feed consumption was registered between medicated and non‐medicated marine feed, however a difference was noted between the medicated and non‐medicated salmonid feed.     

Experimental treatments with diflubenzuron and deltamethrin of sea bass, Dicentrarchus labrax L., infected with the isopod, Ceratothoa oestroides

Sea bass with approximate average weights of 5 and 20 g were treated against Ceratothoa oestroides infection with: (i) medicated pellets of diflubenzuron PC90 at a dosage of 3 mg kg^?1 body weight (BW) per day for 14 days. Lice were counted at the beginning of treatment and 19 days after treatment. The drug cleared all lice in the treated group; in the control group, infection remained high 30 days after beginning the experiment. It was concluded that medicated pellets containing 3 mg kg^?1 BW diflubenzuron effectively cleared pre‐adult and adult stages of the isopod parasite over a 14‐day period. No adverse effects were recorded in treated sea bass during the trials and no reinfection occurred 15 days after end of the treatment. (ii) Deltamethrin by means of bath treatments in infected sea bass kept in experimental tanks at 20°C. Before treatment, toxicity on healthy fish was preliminarily assessed by testing five fish from each size group at concentrations of 30, 10, 5, 3, 1, 0.1, 0.05 and 0.01 mg L^?1 for 30 min. The therapeutic concentrations tested were: 10, 5, 3, 0.15, 0.1, and 0.05 μg L^?1 and assessed at 1, 24 and 48 h. Best results were achieved with the 10 μg L^?1 (0.01 mg L^?1) dose, where prevalence was reduced from 100 to 0% over 24 h in both large and small fish. No parasite recovery was observed at 48 h. The dose of 5 μg L^?1 reduced prevalence from 100 to 11.7% and to 0% for small and large fish, respectively. Finally, with the 3 μg L^?1 dose, prevalence was reduced from 100 to 37.5% (small fish) and to 13.3% (large fish). Lower doses were ineffective on the parasites at either 24 or 48 h.     

Dietary Exposure to Lead and Cadmium in Sweden

Abstract

A duplicate diet study on male and female pensioners in 1970–71 showed mean daily dietary lead intakes of 30 μg and 19 μg respectively. The corresponding cadmium intakes were 10.5μg and 12.9 μg. Analysis of duplicate diets collected during seven consecutive 24-hour periods from 15 women in Stockholm in 1988 showed a mean daily lead intake of 26 μg (range 13–40 μg). The corresponding cadmium intake was 8.5μg (range 5.7–14 μg). Analysis of faeces samples corresponding to the duplicate diets showed similar lead and cadmium contents (mean lead content 24 μg day^?1, range 10–41 μg day^?1; mean cadmium content 8.9 μg day^?1 range 5.5–12 μg day^?1). The median lead and cadmium concentrations in human milk collected in Uppsala were 2 μg kg^?1 and 0.1 mg kg^?1 respectively. The median weekly intakes of lead and cadmium by the breast-fed infants were calculated to be 2 μg kg^?1 body weight and 0.1 μg kg^?1 body weight. Analysis of seven daily diets, together representing the weekly diet of an adult Swedish male, showed a mean lead content of 26 μg (range 15–45 μg), and a mean cadmium content of 10 μg (range 7–15 μg). The mean daily intakes of lead and cadmium found by analysing market baskets prepared in 1987 were 17μg and 12μg respectively. Calculations based on food balance sheet data and levels of lead and cadmium in individual foods showed mean daily intakes of 30 μg lead and 14 μg cadmium per person.     

Extracts and phenolic compounds from Zuccagnia punctata as fungicide seed protectants for corn

The ethanolic leaf extract (TZP) of Zuccagnia punctata, its ethereal fraction (Eet), 2′,4′‐dihydroxychalcone (DC), 2′,4′‐dihydroxy‐3′‐methoxychalcone (DMC) and 7‐hydroxy‐3′,4′‐dimethoxyflavone (HF) were evaluated as fungicide seed protectants on corn. Microdilution assays on a set of Fusarium strains showed minimum inhibitory concentrations (MICs) of 400–800 μg mL^?1 (TZP), 50–100 μg mL^?1 (Eet), 25–50 μg mL^?1 (DC), 50–100 μg mL^?1 (DMC) and 200–400 μg mL^?1 (HF), with minimum fungicidal concentration (MFC)/MIC = 1. Suspensions of TZP, Eet, DC and DMC at MIC × 20 incorporated to the grains at rates of 1920, 240, 120 and 240 mg dry matter kg^?1 of grain, respectively, increased the elongation of the primary roots (24–44%) and the number of seminal roots (44–50%). TZP also increased the number of secondary roots. HF was phytotoxic. Suspensions of TZP, Eet, DC and DMC suppressed the endogenous grain mycoflora at levels similar to those recorded for a thiram + carbendazim‐based fungicide. Grains treated with TZP (1920 mg kg^?1), Eet (240 mg kg^?1), DC (120 mg kg^?1) and DMC (240 mg kg^?1) stimulated the growth of the seedling root system both because of fungal suppression and hormetic effects in greenhouse curative and preventive assays against Fusarium verticillioides on a sand/soil substrate. Eet and its chalcones also reduced the severity of seedling blight more than the thiram + carbendazim‐based fungicide in preventive assays and led to the same disease severity observed for the fungicide treatment in the curative assays. Our results show that Eet and its chalcones not only were effective seed protectants against F. verticillioides and other seedborne fungi, but also improved the early performance of maize seedlings.     

Immunomodulation property of hexane fraction of leaves of Cinnamomum tamala Linn. in rats

The leaves of Cinnamomum tamala Linn. (CT) (Lauraceae) clinically used in Ayurveda as antidiabetic and diuretic, but no reports are available towards immunomodulating property. Its hexane fraction (CTH) was orally given to rats for 10 days and delayed type of hypersensitivity (DTH), antibody producton against sheep red blood cells (SRBCs), mitotic index in bone marrow cells and concanavalin A (Con A) mediated proliferation of lymphocytes were assessed. Further on 30 days treatment, change in body weight (BW), spleen weight, thymus weight, bone marrow cellularity and hematological changes were observed. It inhibited significantly the DTH response (IC₅₀ 1475 ± 57.19 mg kg^?1 BW), antibody production, suppressed mitotic index in bone marrow cells along with the suppression of lymphocyte proliferation against Con A (IC₅₀ 63.33 ± 1.95 µg mL^?1). In all experiments, cyclophasphamide and dexamethasone had been used as reference drug for in vivo and in vitro studies, respectively. On 30 days treatment, the CTH (800 mg kg^?1 BW and above) significantly suppressed growth rate, increase of spleen and thymus weight and low bone marrow cellularity. In hematological examination, it inhibited total white blood cell and lymphocytes count and increased per cent of polymorphs. Thus, it could be suggested that the fraction possesses immunosuppressive property at doses, higher than 800 mg kg^?1 BW in rats. Copyright © 2010 John Wiley & Sons, Ltd.     

Effect of GnRHa, pimozide and Ovaprim on ovulation and plasma sex steroid hormones in African catfish Clarias gariepinus

Nine groups each of four fish were injected with a single intramuscular dose of the following preparations: Physiological saline (0.9% NaCl) as a control group, 0.5 ml kg⁻¹ Ovaprim, 20 and 40 μg kg⁻¹ BW of GnRHa, 8 and 16 mL kg⁻¹ pimozide tablets and the following combination of GnRHa with pimozide (GP): 20 μg + 4 mg, 30 μg + 8 mg and 40 μg + 16 mg kg⁻¹ BW. The primary oocyte diameter (POD) before hormone administration ranged from 943.3 to 1071.0 μm. The latency periods (LP) were in the range of 9.0 to 12.0 h after injection. The highest ovulation ratio (OR) was observed in groups Ovaprim, GP(30 + 8) and GP(40 + 16). Other treatments were effective for ovulation, the ovulation ratio in Groups G(40) and GP(20 + 4) were significantly higher than G(20) treatment. The ovulation index (OI) was in the range 62 to 77% and showed significant differences among groups. There was no significant difference in fertilization ratio (FR) among Ovaprim, GP(30 + 8) and GP(40 + 16) groups, while there were significant difference between the previous group and G(20) and G(40) groups. Control, P8, P16 showed negative results in all the parameters LP, OED, OR, OI and FR. Levels of sex steroids were analyzed on 6 and 12 h after initiation of treatments. A significant increase in plasma E₂ with GP(30 + 8) injection was observed 6 and 12 h after injection, while there were no significant increase between all the other groups 6 h after injection. Treatments with GP(20 + 4) resulted in a significant increase in plasma T concentration in females compared with control after 6 h. In contrast, plasma T and E₂ concentrations were lower during the combined GP(20 + 4), GP(30 + 8) and GP(40 + 16) after 12 h than after 16 h of injection. The combined treatments (GnRHa + PIM) are better compared with Ovaprim which gave the same results, they have some advantages, such as reliable response and low cost. Ovaprim is more than 3 to 5-fold of the cost of (GnRH + PIM). Therefore, this method could be useful tool for commercial catfish breeders to ensure spawning success.     

Leydig cell receptors for luteinizing hormone releasing hormone and its agonists and their modulation by administration or deprivation of the releasing hormone

Leydig cells isolated from adult rat testes bound ¹²⁵I-labelled luteinizing hormone releasing hormone (LHRH) agonist with high affinity (K_A=1.2 × 10⁹M) and specificity. LHRH and the 3–9 and 4–9 fragments of LHRH agonist competed for binding sites with ¹²⁵I-LHRH agonist but with reduced affinities, whereas fragments of LHRH, and oxytocin and TRH were largely inactive. Somatostatin inhibited binding at high (10^?4M) concentrations but was inactive at 10^?6M and less. Pretreatment of rats for 7 days with 5 μg/day of LHRH agonist reduced binding of ¹²⁵I-LHRH agonist to Leydig cells $\text{in vitro}$ by 25%, whilst inhibition of endogenous LHRH by antibodies for 7 days caused a 40% decrease.     

Mycotoxigenic fungi and mycotoxins associated with stored maize from different regions of Lesotho

Samples of stored maize from villages located in five different agroecological zones (southern lowlands, northern lowlands, Senqu river valley, foothills and mountains) of Lesotho were collected in 2009/10 and 2010/11 and assessed for contamination with toxigenic fungi. The water activity of all samples collected during the two seasons was <0.70. The total fungal populations of the maize from different regions in the two seasons was not significantly different (p?>?0.05). Fusarium verticillioides, F. proliferatum and F. subglutinans predominated in different regions in both seasons based on molecular analyses. In the 2009/10 season, the isolates of these species all produced FB₁, while in the 2010/11 season, very few produced FB₁. A. flavus isolates (2009/10) were recovered from mountains and Senqu river valley samples while the 2010/11 isolates were predominantly from the foothills and northern lowlands. The mountain isolates of Aspergillus section Flavi produced the highest levels of AFB₁ (20 mg kg^?1). Aspergillus parasiticus was only isolated from the foothills, Senqu river valley and southern lowlands samples, and the AFB₁ levels produced ranged from ‘none detected’ to 3.5 mg kg^?1. The Aspergillus ochraceous isolates were least frequently encountered in both seasons. In the 2009/10 season, the isolates from the northern lowlands produced ochratoxin A (OTA) in culture. No isolates of A. niger from different regions in both seasons produced any OTA. Multi-mycotoxin analyses of the maize samples were done for a range of mycotoxins. At least one sample from each region in both seasons was FB₁-positive. FB₁ levels for 2010/11 samples (7–936 μg kg^?1) were higher than in the 2009/10 season (2–3 μg kg^?1). In both seasons, the mountains registered the highest levels of FB₁. Deoxynivalenol (DON) was recovered from all the samples analysed, with the highest mean contamination of 1,469 μg kg^?1 in samples from the northern lowlands. Moniliformin (MON) was detected from all agroecological zones in the two seasons (5–320 μg kg^?1 in 2009/10; 15–1,205 μg kg^?1 in 2010/11). Emerging toxins such as fusaproliferin (FUS) and beauvericin (BEA) were also detected. OTA was not detected in any of the samples analysed. Only one 2009/10 sample in the Senqu river valley was positive for AFB₁. This is the first report on toxigenic fungi and multi-mycotoxin contamination of maize samples from subsistence farmers’ stores in different agroecological zones of Lesotho.     

Effects of the Crotalus durissus terrificus snake venom on hepatic metabolism and oxidative stress

Snake venoms present different action mechanisms because of their complex composition, represented mainly by toxins and enzymes. This work aimed to investigate the effects of the Crotalus durissus terrificus(Cdt) venom in the liver. Wistar rats were inoculated intraperitoneally with saline (control) or Cdt venom. After 3, 4, or 6 h, the following parameters were analyzed: (a) hepatic function, (b) oxidative stress parameters, and (c) the metabolism of alanine in the isolated perfused liver. Plasma activities of alanine aminotransferase and aspartate aminotransferase and hepatic glutathione S‐transferase and catalase presented significant elevation in rats inoculated with 300 μg ? kg^?1 Cdt venom. Liver lipoperoxidation was enormously increased by venom doses of 100, 200, and 300 μg ?kg^?1, whereas glutathione S‐transferase was not changed. Perfused livers from rats inoculated with 1500 μg ?kg^?1 venom showed increased production of lactate, pyruvate, and ammonia when alanine was the metabolic substrate. These results demonstrate that the Cdt venom can produce several changes in hepatocytes. The causes of the changes are possibly related to the disequilibrium in the redox homeostasis but also to specific needs of the poisoned organism, for example, an increased supply of lactate and pyruvate in response to an increased activity of the Cori cycle. © 2010 Wiley Periodicals, Inc. J Biochem Mol Toxicol 25:195–203, 2011; View this article online at wileyonlinelibrary.com . DOI 10.1002/jbt.20376     

The effects of water temperature and hormone treatments on circulating LH and ovulation in tench (Tinca tinca)

The effectiveness of three hormone treatments commonly used for artificial reproduction of tench was evaluated under three thermal regimes, with focus on the need for dopamine antagonist. Mature females were divided into nine groups (n?=?8) and gradually exposed, over a 24?h period, to three temperature regimes: cold (18.1?±?0.02?°C), optimal (22.05?±?0.03?°C), and warm (26.3?±?0.01?°C). Each temperature regime comprised three experimental groups injected with one of three hormone treatments: carp pituitary extract (CPE; 3?mg?kg^?1); [D-Arg⁶, Pro⁹, NEt]-sGnRH (10???g?kg^?1); and [D-Arg⁶, Pro⁹, NEt]-sGnRH (10???g?kg^?1)?+?metoclopramide (20?mg?kg^?1) (combined treatment). No differences were found between ovulation induction (ovulation rate????75?%) with sGnRHa alone and with the combined treatment; whereas CPE at cold and warm water temperatures was significantly less effective (P?<?0.05) than above mentioned treatments. Administration of sGnRHa alone induced a gradual increase in luteinizing hormone (LH) levels with LH peaks close to ovulation, in contrast to the immediate LH surge with high LH levels throughout the entire study observed with the combined treatment under all thermal regimes. LH levels induced by GnRHa alone were significantly lower (P?<?0.05) at all temperatures compared to the combined treatment, with the exception of the final sample at 26?°C, when no difference was recorded. Based on these results we recommend the application of sGnRHa without the addition of dopamine antagonist as a reliable method for inducing ovulation in tench under suboptimal temperature conditions.     

Stereological estimation of the surface area and oxygen diffusing capacity of the respiratory stomach of the air‐breathing armored catfish Pterygoplichthys anisitsi (Teleostei: Loricariidae)

The stomach of Pterygoplichthys anisitsi has a thin, translucent wall and a simple squamous epithelium with an underlying dense capillary network. In the cardiac and pyloric regions, most cells have short microvilli distributed throughout the cell surface and their edges are characterized by short, densely packed microvilli. The mucosal layer of the stomach has two types of pavement epithelial cells that are similar to those in the aerial respiratory organs. Type 1 pavement epithelial cells, resembling the Type I pneumocyte in mammal lungs, are flat, with a large nucleus, and extend a thin sheet of cytoplasm on the underlying capillary. Type 2 cells, resembling the Type II pneumocyte, possess numerous mitochondria, a well‐developed Golgi complex, rough endoplasmic reticulum, and numerous lamellar bodies in different stages of maturation. The gastric glands, distributed throughout the mucosal layer, also have several cells with many lamellar bodies. The total volume (air + tissue), tissue, and air capacity of the stomach when inflated, increase along with body mass. The surface‐to‐tissue‐volume ratio of stomach varies from 108 cm^?1 in the smallest fish (0.084 kg) to 59 cm^?1 in the largest fish (0.60 kg). The total stomach surface area shows a low correlation to body mass. Nevertheless, the body‐mass‐specific surface area varied from 281.40 cm² kg^?1 in the smallest fish to 68.08 cm² kg^?1 in the largest fish, indicating a negative correlation to body mass (b = ?0.76). The arithmetic mean barrier thickness between air and blood was 1.52 ± 0.07 μm, whereas the harmonic mean thickness (τ_h) of the diffusion barrier ranged from 0.40 to 0.74 μm. The anatomical diffusion factor (ADF = cm² μm^?1 kg^?1) and the morphological O₂ diffusion capacity (D_morpholO₂ = cm³ min^?1 mmHg^?1 kg^?1) are higher in the smallest specimen and lower in the largest one. In conclusion, the structure and morphometric data of P. anisitsi stomach indicate that this organ is adapted for oxygen uptake from air. J. Morphol. 2009. © 2008 Wiley‐Liss, Inc.     

Dietary total aromatic amino acid requirement and tyrosine replacement value for phenylalanine in Indian major carp: Cirrhinus mrigala (Hamilton) fingerlings

Two 8‐week growth trials were conducted to determine total aromatic amino acid requirement and tyrosine replacement value for phenylalanine in Cirrhinus mrigala fingerlings. To determine the phenylalanine requirement, 20 fish were randomly stocked in triplicate groups in 55‐L indoor polyvinyl flow‐through circular tanks and fed six experimental diets containing graded levels of phenylalanine (5.0, 7.5, 10.0, 12.5, 15.0 and 17.5 g kg^?1, dry diet) with 10 g kg^?1 tyrosine. Maximum weight gain (287%), best FCR (1.44) and PER (1.74) occurred at 12.5 g kg^?1 dietary phenylalanine. Quadratic regression analysis of weight gain, FCR and PER data indicated phenylalanine requirement at 13.5, 12.9 and 12.7 g kg^?1 of dry diet, respectively. Protein deposition was significantly (P < 0.05) higher at 12.5 g kg^?1 dietary phenylalanine. Based on the above results, phenylalanine requirement of C. mrigala is recommended at 13.0 g kg^?1 of dry diet, corresponding to 32.5 g kg^?1 of protein. On the basis of the above requirement, a second experiment with a similar design was conducted using six diets containing graded levels of tyrosine (2.1, 4.0, 6.0, 8.0, 10.0 and 12.0 g kg^?1) with 13.0 g kg^?1 phenylalanine fixed in all diets to determine the phenylalanine replacement value with that of tyrosine. Maximum weight gain (315%), best FCR (1.47) and PER (1.69) was at 8.0 g kg^?1 dietary tyrosine. Quadratic regression analysis of weight gain, FCR and PER data indicated tyrosine requirement at 9.0, 8.4 and 8.2 g kg^?1 of dry diet, respectively. Protein deposition was significantly (P < 0.05) higher at 8.0 g kg^?1 dietary tyrosine. On the basis of the above results, 8.5 g kg^?1 tyrosine, corresponding to 21.3 g kg^?1 of protein, is taken as the optimum requirement and the replacement value is 39.53% on a weight and 36% on a molar basis. Thus, the total aromatic amino acid requirement is 21.5 g kg^?1 of diet, corresponding to 53.8 g kg^?1 of protein for optimum C. mrigala growth.     

Multi-Residue Analysis of Herbicides in Soil with an UPLC-ESI-MS Method

A rapid and sensitive method was developed for the determination of 51 herbicides in soil by ultra-performance liquid chromatography-electrospray ionization-mass spectrometry (UPLC–ESI–MS). Using acetonitrile effectively extracted 22 kinds of triazine and other basic herbicides, and using 90:10 v/v acetonitrile-phosphate buffer (pH = 7.5) effectively extracted another 29 herbicides. The extract has not cleaned up further. Chromatographic separation was achieved within 10 min using gradient elution with acetonitrile–water as a mobile phase for 22 triazine and phenylurea herbicides, and with 5 mM ammonium acetate containing 0.1% formic acid aqueous solution–acetonitrile as a mobile phase for another 29 herbicides. The response was linear over two orders of magnitude with correlation coefficients (r²) higher than 0.99. The limits of quantification for the herbicides varied from 0.2 to 20 µg kg^?1. The intra- and inter-day precisions (relative standard deviation, RSD) were 2.2–9.3% and 5.7–17.1%, respectively. The average recovery varied from 61.6 to 112% with the RSD of 1.6–11.3%. Analyzing 51 soil samples from 17 counties formed the basis of this method. Three herbicide residues were found in four counties. Atrazine residue in soil for 17 counties was found; its content was 0.4–9.8 μg kg^?1. Nicosulfuron residue in soil for two counties was found, with a high up to 133 or 1317 μg kg^?1. Propazine (0.3 and 1.34 μg kg^?1), atratone (2.14 and 3.93 μg kg^?1), and cynanazine (0.34 μg kg^?1) in soils for some counties were also found. The validated method can ensure the rapid multi-class, multi-residue analysis at low μg kg^?1 level for 47 herbicides in soil. The developed method provides an effective analytical basis for controlling herbicide dosage, investigating their distribution and degradation, and evaluating their hazards on the environment and human health.     

Simultaneous real‐time assay of copper and cadmium ions by infrared photo diode electrode implanted in the muscle of live fish

The electrical circuit of an infrared photodiode electrode (IPE) was used in the simultaneous assay of copper and cadmium ions. The electrode's cyclic voltammetry (CV), chronoamperometry and square‐wave (SW) stripping voltammetric optimum conditions were examined. Results for 0–160 mg L^?1 and 50–400 μg L^?1 SW Cu(II) Cd(II), the relative standard deviation of 0.158 Cu(II), 0.077 Cd(II) (n = 15) using 20.0 mg L^?1 have been obtained at optimum conditions. The low detection limit (S/N) was attained to be at 14.71 μg L^?1(2.31 × 10^?7 mol L^?1) Cu(II) and 18.42 μg L^?1(1.63 × 10^?7 mol L^?1) Cd(II). The handmade electrode was implanted deep in the muscle of live fish and interfaced with an electrochemical workstation. Real‐time analytical application was performed on the online assay of living tissue as the specimen was moving. The methods are deemed useful in interfaced assay for physiological control, nanodiode fabrication, and in the production of laboratory on a biochip. © 2009 Wiley Periodicals, Inc. J Biochem Mol Toxicol 23:256–262, 2009; Published online in Wiley InterScience ( www.interscience.wiley.com ). DOI 10.1002/jbt.20287     

The geographic variation and potential risk of DDT in the blood of Pied Kingfishers from northern KwaZulu-Natal,South Africa

Evans, S.W. & Bouwman, H. 2000. The geographic variation and potential risk of DDT in the blood of Pied Kingfishers from northern KwaZulu-Natal, South Africa. Ostrich 71 (1 & 2): 351–354.

DDT has, since 1946 been used in the intradomicilliary control of malaria in northern KwaZulu-Natal. The Pied Kingfisher was selected as representative for organisms in relatively high trophic levels. Blood was obtained from Pied Kingfishers at Kosi Bay (n = 5), Pongolo Floodplain (n = 13), Mkuzi Nature Reserve (n = 4), Ndumu Nature Reserve (n = 4) and St Lucia (n = 3), extracted and analysed [SWEl] via gas chromatography. The highest blood DDE and σDDT concentrations were obtained for the birds from the Pongolo Floodplain (means of 95.92 μg 1^?1/107.01 μg μg 1^?1) and Kosi Bay Nature Reserve (means of 189.09 μg 1^?l/241.8 μg 1^?1). DDT was detected in the blood of Pied Kingfishers from Kosi Bay (mean 47.14 μg 1^?1) and Pongolo Floodplain (mean 44.34 pg 1^?1) only. This indicated their proximity to DDT application and the greater influx of DDT and its metabolites into the water component of these systems. The EDDT plasma concentrations in the Pied Kingfisher blood were calculated by multiplying the blood values of σDDT by 1.8. Using the regression, log₁₀Y = 0.7785 + 0.8593 (log₁₀X), relating the σDDT in eggs to σDDT in plasma of American Kestrel Falco sparverius it was possible to calculate the mean Pied Kingfisher egg σDDT concentration. The approximate mean Pied Kingfisher egg concentration of σDDT was calculated at 2.26 mg kg^?1 for Kosi Bay and 1.24 mg kg^?1 for the Pongolo Floodplain. Using the highest calculated plasma value of σDDT for Kosi Bay and the Pongolo Floodplain indicated that egg σDDT concentrations could be as high as 4.01 mg kg^?1 and 4.17 mg kg^?1 respectively. These calculated levels may be significant when compared to levels of DDE, known to have a detrimental effect, in the eggs of the Brown Pelican Pelecanus occidentalis, where a concentration of 2.5 to 3 mg kg^?1 was associated with substantially impaired reproductive success. The highest calculated egg concentration was nearing this level and it is therefore possible that the Pied Kingfisher population may be at risk.     

Resistance of the mouse to the antifertility effects of LHRH agonists

In the mouse, the LH-releasing activity of the LHRH agonist, D-Trp⁶-N^α-Meleu⁷-DesGly¹⁰-Pro⁹-NHEt-LHRH (Wy-40, 972), was established by its ability both to induce ovulation when administered at 1600 hours on the second day of diestrus and to elevate serum LH in adult males. While Wy-40, 972 was only slightly less active in terms of these end points than it was in the rat, the predictive and possibly causal association between LH-releasing and antifertility activity established for this LHRH analog in the rat could not be clearly identified in the mouse. A total daily dose of 1000 μg Wy-40, 972/mouse was required to completely inhibit pregnancy during days 1–7 of pregnancy and produced only partial inhibition during days 7–12. This dose represents, on a body weight basis, 8250 times the 100 percent effective pregnancy-terminating dose for the rat during equivalent intervals. The resistance of the mouse to the antifertility activity of Wy-40, 972 was found not to be restricted to this particular LHRH analog or to the reproductive state. Administration of another potent LHRH analog, D-Ala⁶-DesGly¹⁰-Pro⁹-NHEt-LHRH (Wy-18,481), to adult male mice at a dose of 100 μg/mouse/day for up to 14 days had no inhibitory effect on the weights of the testes or sex accesory organs. This dose of Wy-18,481 is 7500 times that necessary for significant reduction of these reproductive organ weights in rats within 7 days of treatment. Investigations as to the nature of the mouse's apparently divergent response to the LHRH agonists may further elucidate the antifertility mechanism of such compounds in susceptible species.     

Testosterone treatment augments LHRH‐a induced ovulation vin greenback flounder

Greenback flounder were treated with either an injection of des Gly¹⁰ [D‐Ala⁶] LHRH ethylamide (LHRH‐a), a silastic pellet implant of testosterone (T) or LHRH‐a+T. LHRH‐a and LHRH‐a+T induced more ovulatory events than in control fish or fish treated with T alone. T treatment did not significantly increase the number of ovulatory events above control levels. Treatment with LHRH‐a+T stimulated more ovulatory events than treatment with LHRH‐a alone, suggesting T increases pituitary responsiveness to LHRH‐a. Plasma levels of 17β‐estradiol (E ₂) were significantly higher in fish treated with LHRH‐a and LHRH‐a+T than in control fish or fish treated with T. Plasma levels of 17,20β‐dihydroxy‐4‐pregnen‐3‐one were not elevated above control levels. Daily ovulations, elevated E ₂ and T levels and presumably elevated GtH‐II levels, persisted well after the expected clearance time of exogenous LHRH‐a. Positive steroid feedback may have contributed to the processes of repeated final oocyte maturation and ovulation.     

Changes in amino acid composition in the tissues of African catfish (Clarias gariepinus) as a consequence of dietary L-carnitine supplements

A study was undertaken to examine the effect of different amounts of dietary lysine (13 and 21 g kg^?1 diet), lipid (80 and 160 g kg^?1 diet) and L ‐carnitine (0.2 and 1.0 g kg^?1 diet) on growth performance, proximate composition and amino acid metabolism of the African catfish (Clarias gariepinus). Juvenile African catfish (23 ± 1.5 g/fish) were stocked into 70‐L aquaria (16 aquaria, 28 fish/aquarium) connected to a recirculation system during a maximum period of 74 days. All groups were fed at a level of 24 g kg^?0.8 day^?1 in an experiment run at pair feeding. Animals receiving 1.0 g carnitine accumulated up to six times more carnitine in their tissues than animals receiving 0.2 g (P < 0.05). Acyl‐carnitine and free L ‐carnitine levels increased in the whole body and in tissues. Dietary L ‐carnitine supplements increased protein‐to‐fat ratios in the body, but did not affect growth rate. Protein‐to‐fat ratios were only affected when the biosynthesis capacity of L ‐carnitine was restricted due to low lysine levels and when there was a shortage of dietary fat. When lysine was offered at 21 g kg^?1 feed, dietary L ‐carnitine supplements did not affect the amino acid concentrations of body tissues. Dietary L ‐carnitine supplements raised the concentration of glutamic acid > aspartic acid > glycine > alanine > arginine > serine > threonine in skeletal muscle tissue (P < 0.05). Total amino acid concentration in muscle and liver tissues (dry‐matter basis) increased from 506 to 564 and from 138 to 166 mg g^?1, respectively, when diets were offered with high L ‐carnitine, low lysine and low fat levels. These data suggest that dietary L ‐carnitine supplementation may increase fatty acid oxidation and possibly decrease amino acid combustion for energy.     

Paper‐Based Energy‐Storage Devices Comprising Carbon Fiber‐Reinforced Polypyrrole‐Cladophora Nanocellulose Composite Electrodes

Composites of polypyrrole (PPy) and Cladophora nanocellulose, reinforced with 8 μm‐thick chopped carbon filaments, can be used as electrode materials to obtain paper‐based energy‐storage devices with unprecedented performance at high charge and discharge rates. Charge capacities of more than 200 C g^?1 (PPy) are obtained for paper‐based electrodes at potential scan rates as high as 500 mV s^?1, whereas cell capacitances of ～60–70 F g^?1 (PPy) are reached for symmetric supercapacitor cells with capacitances up to 3.0 F (i.e.,0.48 F cm^?2) when charged to 0.6 V using current densities as high as 31 A g^?1 based on the PPy weight (i.e., 99 mA cm^?2). Energy and power densities of 1.75 Wh kg^?1 and 2.7 kW kg^?1, respectively, are obtained when normalized with respect to twice the PPy weight of the smaller electrode. No loss in cell capacitance is seen during charging/discharging at 7.7 A g^?1 (PPy) over 1500 cycles. It is proposed that the nonelectroactive carbon filaments decrease the contact resistances and the resistance of the reduced PPy composite. The present straightforward approach represents significant progress in the development of low‐cost and environmentally friendly paper‐based energy‐storage devices for high‐power applications.