Dietary restriction during murine development provides protection against MNU-induced mutations

The developmental stage is the most rapid period for the accumulation of somatic mutations. Epidemiological studies have also suggested a significant role of early life for cancer susceptibility, showing a protective effect of modest dietary restriction early in life. To determine if mutation rate, diet, and cancer risk are related, we have investigated the effect of dietary restriction on somatic mutations early in life. The diet of mouse dams was restricted during pregnancy and lactation by 10% from ad libitum control. F(1) pups (SWRxMutaMouse) were weaned at 3 weeks of age. Pups from dams that were on a restricted diet were kept under dietary restriction (40% until 5 weeks of age and then 20% until sacrifice). Only females from litters of seven or eight were used in this study. A portion of pups from both groups were treated with N-methyl-N-nitrosourea (MNU, 50mg/kg, i.p.) at 5 weeks of age and all mice were sacrificed at 10 weeks of age. The frequency of induced mutations was reduced by about 30% at the three loci studied, lacZ (P=0.028) and cII (P=0.042) and Dlb-1 (P=0.032) in the small intestine in the restricted group. A similar decrease in the lacZ mutant frequency was observed in the bone marrow, but the results did not reach statistical significance (P=0.074). Few differences in the lacZ mutant frequency were observed in the colon and the mammary epithelium, but variability of the mutant frequencies was such that an effect of similar magnitude could not be excluded statistically. Analysis of 47 cII mutants revealed that the majority of MNU-induced mutations were G:C to A:T transition at non-CpG sites, with no difference in the mutation spectrum between the two dietary groups.     

Mutations induced by 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK) in the lacZ and cII genes of Muta™ Mouse

4-(Methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK) found in chewing tobacco, snuff, cigarettes, and cigars is a tobacco-specific nitrosamine and classified as a possible human carcinogen (Class 2B) by the International Agency for Research on Cancer (IARC). NNK given intraperitoneally was seen to induce lung and liver adenomas.To evaluate the genotoxicity of NNK in vivo, NNK was intraperitoneally administered to Muta™ Mouse at two concentrations (125 and 250 mg/kg, once a week for 4 weeks) followed by the measurement of mutant frequencies in the lacZ and cII genes from lung and liver in the same mice. Characterization of the types of the mutation was determined by sequencing the cII genes from mutant plaques. The mutant frequencies in both target genes from both organs dose-dependently increased up to 10 times compared to those of the control group. For the types of mutations, the ratio of the G:C to A:T mutation in the total number of mutants was less than the ratio of A:T to T:A and A:T to C:G transversion, contrary to a previous report [Cancer Res, 49 (1989) 5305]. The A:T to T:A transversion was the most highly induced mutation both in the lung and liver cII genes. The increasing rate of mutant frequencies in lung and liver over the vehicle control was 55 and 56 times, respectively, while the increasing rate of G:C to A:T transition was only 1.9 and 2.8 times, respectively.These observations show that NNK predominantly induces DNA adducts leading to A:T to T:A and/or A:T to C:G mutations in the transgene.     

Maternal treatment with oleoyl-estrone induces resistance to lipid accrual in their descendants

Objective: To determine whether treatment of rat dams with oleoyl‐estrone (OE) has an effect on the offspring's long‐term response to diet restriction during lactation. Methods and Procedures: Control, OE‐treated, and diet‐restricted dams were treated up to day 15 of lactation. Changes in food intake and body weight were recorded for dams and their pups. After weaning, pups received a 4‐week standard diet followed by a 4‐week period of high‐fat diet. Lipid, protein, and energy content of pups plus energy intake and efficiency. Serum metabolites (glucose, urea, and cholesterol) and serum hormones (adiponectin, leptin, insulin, and sexual hormones). Results: Neither pups from dams in the OE‐treated nor in the diet‐restricted group showed significant changes in weight, though these two groups ingested 79% of food ingested by controls. At weaning, the pups from OE‐treated rats were smaller than those of the control or diet‐restricted groups. These pups maintained the differences in size and lipid content during the 4‐week standard‐diet period, whereas pups from diet‐restricted dams showed a sharp decrease in their lipid content. During the 4 weeks of high‐fat diet, the male offspring from OE‐treated dams increased the difference in lipid content in relation to the pups from control dams whereas in females the differences decreased. Female offspring from diet‐restricted dams showed the most marked changes in metabolite and hormone levels in relation to controls. Discussion: Treatment of lactating dams with OE programs the metabolic response of their offspring to resist the challenge of a high‐fat diet that would lead to obesity in adulthood.     

日粮添加褐藻糖胶对断奶仔猪抗炎能力和肠道微生物多样性的影响

【目的】本试验旨在研究日粮添加海带提取物褐藻糖胶对断奶仔猪生长性能、营养物质消化率、机体免疫力和肠道微生物多样性的影响。【方法】试验选用36头初始体重为(7.43±0.12) kg的健康仔猪,按照随机区组设计分为3组,每组12头。日粮处理组分别为不含抗生素的基础日粮组、抗生素组和褐藻糖胶组;试验期为28d。评价褐藻糖胶对仔猪生长性能和营养物质消化率的影响;通过比色法和酶联免疫吸附法检测血清中与免疫相关的指标;通过16S rRNA扩增子高通量测序检测试验第0、14和28天肠道微生物多样性。【结果】日粮添加褐藻糖胶可降低试验0–14 d仔猪耗料增重比(P0.05),但对试验全期仔猪平均日增重和平均日采量无显著影响(P0.05)。与对照组相比,饲喂褐藻糖胶日粮后,仔猪的中性洗涤纤维和酸性洗涤纤维表观消化率显著提高(P0.05);仔猪饲喂抗生素和褐藻糖胶日粮后,血清IL-22含量显著降低。试验第14天,抗生素组和褐藻糖胶处理组中Bacteroidetes数量呈上升趋势(P=0.07);试验第28天,抗生素组和褐藻糖胶处理组Actinobacteria丰度显著高于对照组(P0.05),且褐藻糖胶处理组Bacteroides属的菌群丰度显著高于对照组和抗生素组。【结论】日粮添加褐藻糖胶提高了断奶仔猪纤维养分消化率和拟杆菌属的丰富度和多样性,并且降低了促炎性细胞因子IL-22含量,这有助于缓解仔猪的断奶应激反应,建立稳定健康肠道菌群。     

Dietary protein or energy restriction influences age and weight at puberty in ewe lambs

The working hypothesis was that restricted protein content in diets of prepubertal ewe lambs would delay time of onset of puberty to a greater extent than restricted energy content. Restriction of dietary protein (metabolizable protein, MP) or energy (ME) intake on age and weight at puberty was studied in ewe lambs in two experiments. In Experiment 1, 85 crossbred ewe lambs were assigned by weight to one of five dietary treatment groups: control (CTL; 18 mg ME day⁻¹ and 173 g MP day⁻¹); energy restricted (ER; 10.2 mg ME day⁻¹ and 96 g MP day⁻¹); protein restricted (PR; 11 mg ME day⁻¹ and 66 g MP day⁻¹); ER for 9 weeks then switched to CTL (ER/CTL); PR for 9 weeks then switched to CTL (PR/CTL). Ewe lambs were weighed biweekly, bled weekly, and serum was assayed for progesterone to determine time of puberty. Average age at puberty differed among lambs fed the various diets (P < 0.01). Ewe lambs in the CTL group were younger and weighed more at puberty. Lambs fed diets restricted in protein or energy content were similar in age at puberty. Dietary switching, ER/CTL and PR/CTL, affected body weight and gain (P < 0.05), but did not result in changed age at puberty compared with lambs fed ER or PR diets. In Experiment 2, two breed types (maternal type and meat type) were used to compare responses of dietary protein restriction to dietary energy restriction. In Experiment 2, lambs (n = 60) were assigned to either the CTL, PR or ER diets. Lambs in the CTL group gained more weight than lambs fed the PR diet and lambs fed the PR diet gained more weight than lambs fed the ER diet (P < 0.05). Diet affected age (P < 0.001), but not weight at puberty (P = 0.13). Dietary energy and protein restriction influenced age at puberty in ewe lambs with energy restriction having a greater influence on delaying onset of puberty than protein restriction. Realimentation did not overcome effects of previous feed restriction by inducing an earlier age at puberty than ewe lambs fed the ER or PR diet throughout the experimental period. Body weight gain prepuberty is more important than type of dietary restriction (protein or energy) with regard to impacting age at puberty. We, therefore, reject the working hypothesis that dietary protein restriction prepuberty would have a greater impact in delaying onset of puberty than dietary energy restriction.     

苏云金芽胞杆菌BkdR和CcpA对bkd基因簇的转录调控

【目的】通过分析苏云金芽胞杆菌(Bacillus thuringiensis,Bt)转录调控因子BkdR和多效调控因子CcpA对亮氨酸、异亮氨酸、缬氨酸代谢基因簇bkd的转录调控,明确bkd基因簇的转录调控机制。【方法】通过β-半乳糖苷酶活性测定分析bkd基因簇启动子的诱导转录活性,采用同源重组技术敲除Bt HD73菌株的ccpA基因,通过融合His标签的方法在大肠杆菌中表达纯化BkdR和CcpA蛋白,通过凝胶阻滞实验明确BkdR和CcpA蛋白与bkd基因簇启动子的结合作用。【结果】亮氨酸、异亮氨酸、缬氨酸可诱导bkd基因簇启动子Pptb的转录活性。Pptb的诱导活性在bkdR突变体中明显降低,而在ccpA突变体中明显上升。BkdR和CcpA蛋白与Pptb均有结合作用。【结论】bkd基因簇的转录活性受BkdR正调控,而受CcpA负调控。     

A comparison of the genotoxicity of ethylnitrosourea and ethyl methanesulfonate in lacZ transgenic mice (Muta™Mouse)

We compared the induction of gene mutations and chromosomal aberrations by ethylating agents in lacZ transgenic mice (Muta™Mouse). Chromosomal aberrations were detected by the peripheral blood micronucleus assay. Gene mutations were detected in the lacZ transgene. A small amount of blood was sampled from a tail vessel during the expression time for fixation of gene mutations in vivo; this enabled us to detect and compare clastogenicity and gene mutations in the identical mouse. Single intraperitoneal injections of ENU (50–200 mg/kg) and EMS (100–400 mg/kg) strongly induced micronucleated reticulocytes (MN) detectable in peripheral blood 48 h after treatment. The maximum MN frequencies induced were 6.6% and 3.3% for ENU (100 mg/kg) and EMS (400 mg/kg), respectively (the control value was 0.3%). lacZ mutant frequency (MF) was analyzed in bone marrow and liver 7 days after treatment. Spontaneous MFs were 2.0–4.6x10⁻⁶. MF in bone marrow was increased by ENU to 3.4x10⁻⁵ at 200 mg/kg and induced by EMS to 1.8x10⁻⁵ at 400 mg/kg. In liver, however, both chemicals at their highest doses induced only slight increases in MF. The induction of both micronuclei and lacZ mutations in bone marrow by both ENU and EMS correlated better with O⁶-ethylguanine adducts than with N7-ethylguanine adducts. The mutants (19 for ENU and 12 for EMS) were subjected to DNA sequence analysis. Among EMS-induced mutations, 75% were GC to AT transitions, which were probably caused by O⁶-ethylguanine. Among ENU-induced mutations, in contrast, 40% occurred as AT base pair substitutions (6 AT to TA transversions and 2 AT to GC transitions) (no such mutations were induced by EMS). These results, together with the known reactivity of ENU to thymine suggest that thymine adducts play a significant role in the ENU mutagenesis.     

Chronic Maternal Vitamin B12 Restriction Induced Changes in Body Composition & Glucose Metabolism in the Wistar Rat Offspring Are Partly Correctable by Rehabilitation

Maternal under-nutrition increases the risk of developing metabolic diseases. We studied the effects of chronic maternal dietary vitamin B12 restriction on lean body mass (LBM), fat free mass (FFM), muscle function, glucose tolerance and metabolism in Wistar rat offspring. Prevention/reversibility of changes by rehabilitating restricted mothers from conception or parturition and their offspring from weaning was assessed. Female weaning Wistar rats (n = 30) were fed ad libitum for 12 weeks, a control diet (n = 6) or the same with 40% restriction of vitamin B12 (B12R) (n = 24); after confirming deficiency, were mated with control males. Six each of pregnant B12R dams were rehabilitated from conception and parturition and their offspring weaned to control diet. While offspring of six B12R dams were weaned to control diet, those of the remaining six B12R dams continued on B12R diet. Biochemical parameters and body composition were determined in dams before mating and in male offspring at 3, 6, 9 and 12 months of their age. Dietary vitamin B12 restriction increased body weight but decreased LBM% and FFM% but not the percent of tissue associated fat (TAF%) in dams. Maternal B12R decreased LBM% and FFM% in the male offspring, but their TAF%, basal and insulin stimulated glucose uptake by diaphragm were unaltered. At 12 months age, B12R offspring had higher (than controls) fasting plasma glucose, insulin, HOMA-IR and impaired glucose tolerance. Their hepatic gluconeogenic enzyme activities were increased. B12R offspring had increased oxidative stress and decreased antioxidant status. Changes in body composition, glucose metabolism and stress were reversed by rehabilitating B12R dams from conception, whereas rehabilitation from parturition and weaning corrected them partially, highlighting the importance of vitamin B12 during pregnancy and lactation on growth, muscle development, glucose tolerance and metabolism in the offspring.     

ERKs activation and calcium signaling are both required for VEGF induction by vanadium in mouse epidermal Cl41 cells

Studies were conducted to determine the immunomodulatory effects of high dietary ascorbic acid (vitamin C) on growth, serum concentration, non-specific immune response and disease resistance of a commercially important Asian catfish, Clarias batrachus. Four practical diets were formulated to contain 0, 500, 1000 and 2000 mg ascorbic acid (AA) equivalent/kg diet, supplied as L-ascorbyl-2-polyphosphate (LAPP) and were fed for 2, 4, 6 and 8 weeks. Each diet was fed to triplicate groups of catfish with initial body weight of 15.47± 0.59 g. After 2, 4, 6 and 8 weeks, growth, serum concentration of AA, oxidative respiratory burst, lysozyme and natural hemolytic complement activities, myeloperoxidase (MPO) content and natural haemagglutination titre were measured. Ten numbers of fish in duplicate were challenged with Aeromonas hydrophila to measure the level of protection against aeromoniasis at each one of the assayed times. The results showed that AA concentration in serum correlated positively with those in the diets and reached its saturation level after the time period directly proportional to the increase in dose level. Fish fed AA-supplemented diets showed significantly (p < 0.05) higher specific growth rate after 2 weeks of feeding. The superoxide production was enhanced after 8 weeks of feeding fish at a supplemented dose level of 2000 mg/kg. Similarly, MPO content, haemagglutination titre and alternative complement activity in serum enhanced with the increase of dietary AA levels at different duration of feeding. The lysozyme activity was not affected by the dietary AA treatment. On the other hand, feeding of AA at all concentrations significantly increased percent survival against A. hydrophila challenge after 4 weeks compared to control. The non-specific immune parameters as well as percent survival were enhanced as a result of high AA supply particularly at 500 mg/kg diet, although the increase was not maintained but returned to the initial levels after 4 weeks. These results support the possible use of AA as an immunostimulant at a dose of 500 mg/kg diet for a period of 4 weeks in catfish farming. (Mol Cell Biochem xxx: 25–33, 2005)     

Dietary low linolenic acid compared with docosahexaenoic acid alter synaptic plasma membrane phospholipid fatty acid composition and sodium-potassium ATPase kinetics in developing rats

The objective of this study was to investigate if maternal dietary 20:4n-6 arachidonic acid (AA) and 22:6n-3 compared with adequate or low levels of 18:3n-3 linolenic acid (LNA) increases synaptic plasma membrane (SPM) cholesterol and phospholipid content, phospholipid 20:4n-6 and 22:6n-3 content, and Na,K-ATPase kinetics in rat pups at two and five weeks of age. At parturition, Sprague-Dawley rats were fed semi-purified diets containing either AA + docosahexaenoic acid (DHA), adequate LNA (control; 18:2n-6 : 18:3n-3 ratio of 7.1 : 1) or low LNA (18:2n-6 : 18:3n-39 ratio of 835 : 1). During the first two weeks of life, the rat pups received only their dams' milk. After weaning, pups received the same diet as their respective dams to five weeks of age. No significant difference was observed among rat pups fed the diet treatments for SPM cholesterol or total and individual phospholipid content at two and five weeks of age. Fatty acid analysis revealed that maternal dietary AA + DHA, compared with feeding the dams the control diet or the low LNA diet, increased 20:4n-6 in phosphatidylserine and 22:6n-3 content of SPM phospholipids. Rats fed dietary AA + DHA or the control diet exhibited a significantly increased Vmax for SPM Na,K-ATPase. Diet treatment did not alter the Km (affinity) of SPM Na,K-ATPase in rat pups at two and five weeks of age. It is concluded that dietary AA + DHA does not alter SPM cholesterol and phospholipid content but increases the 22:6n-3 content of SPM phospholipids modulating activity of Na,K-ATPase.     

油菜类型和品种对外来入侵杂草小子虉草的替代控制作用

外来入侵植物小子虉草（Phalaris minor Retz.）是世界公认的冬季农田恶性杂草,掌握农作物对其替代控制作用具有重要的研究价值。前期研究表明,油菜是替代控制小子虉草的优良农作物,然而,目前尚不清楚油菜类型与品种对其控制能力的影响。为此选取与小子虉草同域发生的不同类型（白菜型油菜、芥菜型油菜和甘蓝型油菜）油菜品种各3种,通过田间小区实验和室内化感作用测定,对比研究其对小子虉草的生长、繁殖、表型以及化感作用的影响。田间实验显示：竞争方式（种内或种间竞争）和油菜类型对小子虉草的地上生物量、种子数、株高、分枝数、叶面积和比叶面积存在极显著（P=0.0001）影响;而油菜品种对小子虉草的地上生物量（P=0.6064）、种子数（P=0.3577）、株高（P=0.4279）、分枝数（P=0.6357）、叶面积（P=0.8839）和比叶面积（P=0.3424）均无显著影响。3种类型油菜对小子虉草生长、繁殖以及表型的影响存在明显差异,其中芥菜型油菜对小子虉草的上述指标的影响最强,而白菜型油菜的影响最弱。室内生物测定显示,油菜对小子虉草具有化感抑制作用,当供试油菜叶片水提液浓度为0.1 g/mL时,小子虉草种子的萌发和幼苗的株高、根长、生物量均被显著抑制;研究也表明不同类型油菜对小子虉草的化感作用显著不同,同等条件下,芥菜型油菜对小子虉草的化感抑制作用最强。综上所述,油菜类型对外来入侵小子虉草的控制作用存在显著差异,其中芥菜型油菜对植物小子虉草的替代控制作用明显优于白菜型油菜和甘蓝型油菜,而其强的化感抑草特性或许是其强控草能力的原因之一。另外,本研究也为进一步利用油菜替代控制入侵植物小子虉草提供了参考。     

aubergine mutations in Drosophila melanogaster impair P cytotype determination by telomeric P elements inserted in heterochromatin

Transposable P elements inserted in the heterochromatic Telomeric Associated Sequences on the X chromosome (1A site) of Drosophila melanogaster have a very strong capacity to elicit the P cytotype, a maternally transmitted condition which represses P element transposition and P-induced hybrid dysgenesis. This repressive capacity has previously been shown to be sensitive to mutant alleles of the gene Su(var)205, which encodes HP1 (Heterochromatin Protein 1), thus suggesting a role for chromatin structure in repression. Since an interaction between heterochromatin formation and RNA interference has been reported in various organisms, we tested the effect of mutant alleles of aubergine, a gene that has been shown to play a role in RNA interference in Drosophila, on the repressive properties of telomeric P elements. Seven out of the eight mutant alleles tested clearly impaired the repressive capacities of the two independent telomeric P insertions at 1A analyzed. P repression by P strains whose repressive capacities are not linked to the presence of P copies at 1A were previously found to be insensitive to Su(var)205; here, we show that they are also insensitive to aubergine mutations. These results strongly suggest that both RNA interference and heterochromatin structure are involved in the establishment of the P cytotype elicited by telomeric P elements, and reinforce the hypothesis that different mechanisms for repression of P elements exist which depend on the chromosomal location of the regulatory copies of P.Communicated by G. Reuter     

Does Perinatal ω‐3 Polyunsaturated Fatty Acid Deficiency Increase Appetite Signaling?

Objective: To investigate the effect of maternal dietary ω‐3 polyunsaturated fatty acid (PUFA) deficiency and repletion on food appetite signaling. Research Methods and Procedures: Sprague‐Dawley rat dams were maintained on diets either supplemented with (CON) or deficient in (DEF) ω‐3 PUFA. All offspring were raised on the maternal diet until weaning. After weaning, two groups remained on the respective maternal diet (CON and DEF groups), whereas a third group, born of dams fed the DEF diet, were switched to the CON diet (REC). Experiments on food intake began when the male rats reached 16 weeks of age. Food intake was stimulated either by a period of food restriction, by blocking glucose utilization (by 2‐deoxyglucose injection), or by blocking β‐oxidation of fatty acids (by β‐mercaptoacetate injection). Results: DEF animals consumed more than CON animals in response to all stimuli, with the greatest difference (1.9‐fold) demonstrated following administration of 2‐deoxyglucose. REC animals also consumed more than CON animals in response to food restriction and 2‐deoxyglucose but not to β‐mercaptoacetate. Discussion: These findings indicate that supply of ω‐3 PUFA, particularly during the perinatal period, plays a role in the normal development of mechanisms controlling food intake, especially glucoprivic (i.e. reduced glucose availability) appetite signaling. Dietary repletion of ω‐3 PUFA from 3 weeks of age restored intake responses to fatty acid metabolite signaling but did not reverse those in response to food restriction or glucoprivic stimuli.     

Population Genetic Structure of Two Columnar Cacti with a Patchy Distribution in Eastern Brazil

The genetic variability and population genetic structure of six populations of Praecereus euchlorus and Pilosocereus machrisii were investigated. The genetic variability in single populations of Pilosocereus vilaboensis, Pilosocereus aureispinus, and Facheiroa squamosa was also examined. All of these cacti species have a patchy geographic distribution in which they are restricted to small areas of xeric habitats in eastern Brazil. An analysis of genetic structure was used to gain insights into the historical mechanisms responsible for the patchy distribution of P. euchlorus and P. machrisii. High genetic variability was found at the populational level in all species (P=58.9–92.8%, A_p=2.34–3.33, H_e=0.266–0.401), and did not support our expectations of low variability based on the small population size. Substantial inbreeding was detected within populations (F_IS=0.370–0.623). In agreement with their insular distribution patterns, P. euchlorus and P. machrisii had a high genetic differentiation (F_ST=0.484 and F_ST=0.281, respectively), with no evidence of isolation by distance. Accordingly, estimates of gene flow (N_m) calculated from F_ST and private alleles were below the level of N_m=1 in P. machrisii and P. euchlorus. These results favored historical fragmentation as the mechanism responsible for the patchy distribution of these two species. The genetic distance between P. machrisii and P. vilaboensis was not compatible with their taxonomic distinction, indicating a possible local speciation event in this genus, or the occurrence of introgression events.     

Ribosomal and chloroplast DNA restriction site mutations and the radiation ofRobinsonia (Asteraceae: Senecioneae) on the Juan Fernandez Islands

Restriction site mutations in the chloroplast (cpDNA) and ribosomal DNA (rDNA) were examined in 41 populations representing five of the seven recognized species of the genusRobinsonia, which is endemic to the Juan Fernandez Islands. No intraspecific variation was detected for cpDNA but one population of one of the species (R. evenia) had a restriction site mutation in rDNA not detected elsewhere. No restriction site mutations were unique to all species ofRobinsonia relative to the species ofSenecio used as outgroups. All 13 mutations (eight from cpDNA and five from rDNA) are restricted to single species, and thus provide no cladistically useful information within the genus. The distribution of mutations is concordant with the hypothesis of a rapid adaptive radiation ofRobinsonia subsequent to the dispersal of its ancestor to Masatierra.     

Emergence of a mutagenic ochre suppressor mutation under lactose selection in appm mutant ofEscherichia coli harbouring the F′lacZU118 episome

WhenEscherichia coli harbouring theppm (earlier calledadi) mutation and the F′lacZU118 episome is subjected to lactose selection in the presence of suboptimal concentrations of glycerol, Lac⁺ colonies emerge after 5–6 days. They are shown to harbour an ochre suppressor mutation at 15.15 min. Inactivation ofrecA results in approximately four-fold reduction in the response. In theppm — ochre suppressor double mutant background the leakiness of thelacZ allele carried by F′ CC105 is enhanced, suggesting misreading of a valine codon (GUG) as glutamic acid codon (GAG). This is accompanied by reversion of thelacZ mutation tolacZ ⁺ (GTG → GAG). In LB medium both the leakiness and reversion are inhibited by streptomycin. Inactivation ofrecA did not affect leakiness but abolished reversion. These data are discussed in relation to the importance of allele leakiness and restricted growth in stationary-phase (adaptive) mutagenesis.     

The effect of Euglena viridis on immune response of rohu, Labeo rohita (Ham.)

The study evaluated the effect of dietary doses of Euglena viridis on the immune response and disease resistance of Labeo rohita fingerlings against infection with the bacterial pathogen Aeromonas hydrophila. L. rohita fingerlings were fed with diet containing 0 (Control), 0.1 g, 0.5 g, 1.0 g Euglena powder kg⁻¹ dry diet for 90 days. Biochemical (serum total protein, albumin, globulin, albumin:globulin ratio), haematological (WBC, RBC, haemoglobin content) and immunological (superoxide anion production, lysozyme, serum bactericidal activity) parameters of fish were examined after 30, 60 and 90 days of feeding. Fish were challenged with A. hydrophila 90 days post-feeding and mortalities were recorded over 10 days post-infection. The results demonstrate that fish fed with Euglena showed increased levels of superoxide anion production, lysozyme, serum bactericidal activity, serum protein and albumin (P < 0.05) compared with the control group. Following challenge with A. hydrophila less survivability was observed in the control group (56.65%) than the group fed the experimental diets. The group fed 0.5 g Euglena kg⁻¹ dry diet showed the highest percentage survival (75%). These results indicate that Euglena stimulates the immunity and makes L. rohita more resistant to A. hydrophila infection.     

Expression of the<Emphasis Type="Italic"> lacZ</Emphasis> reporter gene in sporophytes of the seaweed<Emphasis Type="Italic"> Laminaria japonica</Emphasis> (Phaeophyceae) by gametophyte-targeted transformation

The seaweed Laminaria japonica (Phaeophyceae) has a two-generation life cycle consisting of haploid gametophytes and diploid sporophytes. Female and/or male gametophytes were transformed using particle bombardment and the histological LacZ assay was performed on sporophytes generated by either parthenogenesis or inbreeding. Female gametophyte-targeted transformation resulted in similar lower efficiencies in both parthenogenetic and zygotic sporophytes, and only a chimeric expression pattern was observed. Male gametophyte-targeted transformation led to a higher efficiency, with 3.5% of the zygotic sporophytes stained completely blue (all-blue), implying the integration of lacZ at the one-cell stage. Polymerase chain reaction analysis using primers specific for a lacZ-vector juncture fragment and subsequent blotting indicated the presence of the introduced gene in the sporophytes. The method reported here has a potential for seaweed transformation using spore-based bombardment followed by the developmental process.Abbreviations DPR Detected positive rate - ER Expression rateCommunicated by F. Sato     

Effect of dietary selenium supplementation on resistance to baculovirus infection

We have examined the effects of dietary selenium (Se) supplementation on larval growth and immunocompetence of the lepidopteran pest, the cabbage looper, Trichoplusia ni. Supplementation of the diet of T. ni larvae with 10–20 ppm Se resulted in a 1 day delay in pupation. The effects of the addition and/or removal of dietary Se on total Se bioaccumulation and sequestration were determined by neutron activation analysis of pupae. Early penultimate instar larvae moved from selenium containing diet to basal diet lost total pupal Se content down to the level of those fed basal diet. Conversely, larvae moved from basal diet to diet containing additional Se rapidly attained pupal Se levels comparable to larvae fed Se throughout larval development. Therefore, dietary Se is rapidly accumulated or lost during larval development, but significant amounts are sequestered from diet into pupae. Larvae were reared on diet supplemented with 5 or 10 ppm Se until the onset of the penultimate instar then infected per os with increasing concentrations of the fatal baculovirus Autographa californica multiple nucleopolyhedrovirus (AcMNPV). Larvae fed Se in the penultimate and ultimate instars were more resistant to viral infection than larvae not fed Se in the final instars. This study indicates that dietary Se levels rapidly impact Se assimilation and sequestration and that tissue Se levels are an important factor in resistance to AcMNPV infection in larval T. ni.     

3种无脊椎动物对近江牡蛎Crassostrea ariakensis和熊本牡蛎C. sikamea的捕食研究

捕食是影响牡蛎种群建立和牡蛎礁发育的重要生物因子之一。通过室内受控实验测定了日本蟳（Charybdis japonica）、脉红螺（Rapana venosa）和黄口荔枝螺（Thais luteostoma）对4组规格（W1：壳高10-20mm;W2：壳高20-30mm;W3：壳高30-40mm;W4：壳高>40mm）近江牡蛎（Crassostrea ariakensis）和熊本牡蛎（C.sikamea）的捕食偏好性和捕食效率。双因子方差分析结果表明,日本蟳对2种牡蛎的捕食效率没有显著性差异（P>0.05）,但牡蛎规格大小显著影响着日本蟳的捕食效率（P<0.05）,即日本蟳对W1组近江牡蛎的捕食效率显著高于W2和W4组（P<0.05）,W3组的被捕食效率介于中间（P>0.05）;日本蟳对W1组熊本牡蛎的捕食效率显著高于W2和W3组（P<0.05）,W4组的被捕食效率与其他处理组均没有显著性差异（P>0.05）。牡蛎种类（P=0.590）和规格大小（P=0.357）对脉红螺的捕食效率均无显著性影响,不同规格的两种牡蛎均呈现较低的被捕食效率。黄口荔枝螺对2种牡蛎的捕食效率无显著性差异（P=0.917）,但牡蛎规格大小显著影响其捕食效率（P=0.035）,即对W1组熊本牡蛎捕食效率显著高于其他3个规格组（P<0.05）,但其对不同规格近江牡蛎的捕食效率没有显著性差异（P>0.05）。2种牡蛎的壳厚与其壳高之间均存在极显著的正相关关系（P<0.001）。研究结果表明,3种无脊椎动物捕食者对近江牡蛎和熊本牡蛎并未表现出差异性的捕食偏好,但对不同规格牡蛎的捕食效率具有种间差异。