Presence of Panulirus argus virus 1 (PaV1) in juvenile spiny lobsters Panulirus argus from the Caribbean coast of Mexico

Macroscopic evidence, histological sections, transmission electron microscopy (TEM) evaluation, and PCR analyses of 25 apparently diseased juvenile spiny lobsters Panulirus argus from the reef lagoon of Puerto Morelos, Mexico, showed the presence of Panulirus argus Virus 1 (PaV1). Cowdry Type A intranuclear viral inclusions were observed in histological analyses, icosahedral viral particles were observed by TEM, and PCR using specific primers for PaV1 amplified a fragment of 499 bp. This is the first report of PaV1 infecting P. argus outside the Florida Keys, USA.     

California Condors and DDE: a re-evaluation

Eggshells of wild California Condors Gymnogyps californianus were much thinner in the 1960s, when DDT was used heavily, than during earlier pre-DDT and later reduced-DDT periods. However, eggshell thickness was more strongly linked to egg size (mass) than to measured levels of p,p'DDE (the primary metabolite of DDT). Egg size was consistent within individual females and yielded correlation coefficients with shell thickness ranging from 0.49 to 0.97, depending on the period and the analysis assumptions used. Measured DDE levels, although often substantial, provided only a weak correlation ( r  = −0.33) with shell thickness. In part, the absence of a strong DDE/thickness correlation may have been an artefact of losses of DDE from fragment membranes over time. Nevertheless, the extreme (28–29%) shell thinning of the 1960s was not linked with clearly increased egg-breakage or nest-failure rates, and one female of the 1980s with 25.6% shell thinning was the most productive female of her era. Some eggs with over 30% shell thinning hatched successfully, and broken eggs closely resembled hatched eggs in shell thickness, strongly suggesting that shell thinning was not an important cause of breakage. The apparent absence of harmful effects from the extreme shell thinning of the 1960s may have resulted from (1) the fact that historic pre-DDT condor eggs were on average 16.7% thicker shelled for their mass than predicted by the overall egg mass/shell thickness curve for birds, and (2) a possible egg-size decline or sampling bias toward small-egged females in the 1960s. That DDE was an important cause of the Condor's decline appears unlikely from overall available data.     

Effect of cytochalasins B and D on the developmental competence of somatic cell nuclear transfer embryos in miniature pigs

In many animals, cytochalasins have generally been used as cytoskeletal inhibitors for the diploid complement retention of somatic cell nuclear transfer (SCNT) embryos. However, limited information is available on the effects of cytochalasins on the in vitro development of SCNT embryos. Hence, we compared the effects of cytochalasin B (CB) and cytochalasin D (CD) on pseudo-polar body (pPB) extrusion, cortical actin filament (F-actin) distribution in porcine parthenogenetic oocytes and in vitro development of SCNT embryos that were reconstructed using foetal fibroblasts in the G0/G1 phase derived from miniature pigs. CB (7.5 microg/ml) and CD (2.5 microg/ml) treatments effectively inhibited pPB extrusion in SCNT embryos. CB (2.5 microg/ml) treatment could not inhibit pPB extrusion and insufficiently destabilized F-actin immediately following artificial activation. In parthenogenetic oocytes treated with 2.5 microg/ml CD, normal reorganization and uniform distribution of cortical F-actin at the cytoplasmic membrane were observed at 8 h after artificial activation; this finding was similar to that of control oocytes. In contrast, parthenogenetic oocytes treated with 7.5 microg/ml CB showed non-uniform distribution of F-actin at 8 h after artificial activation. On day 5 after in vitro cultivation, the blastocyst formation rate of SCNT embryos treated with 2.5 microg/ml CD was significantly higher than that of SCNT embryos treated with 2.5 and 7.5 microg/ml CB (p < 0.05). Hence, the present findings suggest that CD is more effective than CB as the cytoskeletal inhibitor for the production of SCNT embryos in miniature pigs.     

Influence of estrogen administration on the growth response to growth hormone (GH) in GH-deficient mice

In women who are growth hormone (GH) deficient, exogenous estrogens increase the dosage of GH that is needed to normalize circulating levels of insulin-like growth factor (IGF-1). Serum IGF-1 derives mostly from the liver, and it is unknown whether the peripheral effects of GH are also impaired by estrogens. Because the ultimate effect of GH is longitudinal growth, we have investigated the influence of estrogen administration on the growth response to recombinant mouse GH therapy in prepubertal GH-deficient (GHD) GHRH knockout (GHRHKO) female mice. Twenty-four GHRHKO female mice (4 animals/group) were treated for 4 weeks (from the second to sixth week of age) with the following schedules: Group I, GH only (25 microg/day); Group II, subcutaneous (sc) ethynil estradiol (EE) (0.035 ES01247g/day); Group III, GH + scEE; Group IV, oral (po) EE (0.035 microg/day); Group V, GH + poEE; Group VI, placebo. At the end of the treatment period, we measured uterine weight, total body weight (TBW), body length (nose-anus, N-A), and femur length. In addition, serum IGF-1 levels were measured. Uteri of mice treated with oral or scEE showed similar increases in weight. There was no difference in the increase in longitudinal growth parameters between mice treated with GH alone or with GH in association with oral or scEE. Serum IGF-1 decreased in animals treated with GH + scEE, compared with GH group, but no group was significantly different from placebo. These results show that subcutaneous or oral EE does not reduce the growth response to GH in female GHD mice.     

金钱鱼毒腺cDNA表达文库的构建及EST序列分析

以金钱鱼 (Scatophagusargus)毒腺为出发材料 ,构建以真核表达载体pcDNA3 0为基础的金钱鱼毒腺cDNA表达文库 .应用SMARTTM cDNALibraryConstruction技术和生物信息学分析等方法 ,通过对文库克隆的序列测定和初步生物信息学分析 ,获得了 2 0 1个金钱鱼新表达序列标签 (ESTs) ,其中已确定全长cDNA的克隆有 2 7个 ,包括多个核糖体大小亚基蛋白 (ribosomalprotein)、细胞凋亡相关蛋白 (programmedcelldeath 10 ) ,G蛋白信号调控子 (Gproteinsignalingregulator)、胸腺素(thymosinbeta 4 )、延伸因子 (translationelongationfactor 1 alpha)和泛素 (ubiquitin)等 .金钱鱼毒腺cDNA表达文库的成功构建 ,为研究金钱鱼毒腺的活性组分及其作用机制奠定了基础 ,也是分离新基因不可多得的重要资源     

Dispersal of the lycaenid Plebejus argus in response to patches of its mutualist ant Lasius niger

We examined the movement rates of individuals of an obligate myrmecophilous butterfly, Plebejus argus (Lycaenidae: Lepidoptera), in relation to a patch of its mutualist ant, Lasius niger . Marked male and female P. argus from local (resident), fragmented and continuous populations were released into and away from (>100m) a patch of L. niger , and subsequently searched for in a 500×500 m area. Recaptures of males and females from all populations were strongly associated with the target patch of mutualist ants. First day recapture displacements were significantly higher for releases away from the target patch for males, but not for females. Mutualist ant presence was shown to be more important than nectar source density in determining persistence of P. argus in release squares. There was no significant difference in displacement distances or recapture frequencies (standardised for search effort) in the target patch of individuals from local (resident of target patch), fragmented and continuous populations. The recapture frequency in the target patch of males from the local population released outside the target patch was greater than expected from a null model of random straight line movement, suggesting non-random (systematic or orientated) search behaviour. The number of males remaining in the target patch following release into the target patch was greater than expected from a simple diffusion model, suggesting a behavioural response to patch boundaries or other cues related to the patch.     

DDT对多刺裸腹溞的急性毒性和生命表参数的影响

采用48 h急性毒性试验研究了DDT对多刺裸腹溞的48小时LC₅₀值,采用生命表试验方法研究了暴露于不同浓度（1、8、16、24、32和40 μg·L^-1）DDT溶液中的多刺裸腹溞的生命表统计学参数.结果表明：DDT对多刺裸腹溞48小时LC₅₀值为324 μg·L^-1;浓度在1～40 μg·L^-1的 DDT对多刺裸腹溞的生命期望、净生殖率、世代时间和总繁殖率均没有显著影响(P>0.05),但对种群内禀增长率(r_m)影响显著 (P<0.05).与空白对照组相比,浓度为8、16和40 μg·L^-1的DDT显著地提高了多刺裸腹溞种群r_m.在使用多刺裸腹溞的生命表统计学参数监测亚致死浓度的DDT的生态影响时,r_m是较敏感的指标.     

Androgen and estrogen stimulation of ornithine decarboxylase activity in mouse kidney

In the present work, the activity of mouse renal ornithine decarboxylase (ODC) from CBA female mice was used as a biological marker to detect (anti)androgenic activity of different groups of endocrine disruptors and steroids. Daily injections of testosterone or dihydrotestosterone (DHT) into 60 day old female mice for 4 days increased renal ODC activity in a dose-dependent manner that reached up to 100-fold (testosterone) or 250-fold (DHT) above the baseline when the highest dose, 200 microg/mouse, was used. Administration of flutamide concurrently with testosterone (75 microg/mouse) caused a potent decrease of ODC induction in a dose-dependent manner, suppressing the enzyme activity at the doses of 0.1 and 0.5 mg/mouse by about 88 and 95%, respectively. In contrast, estradiol at the doses of 0.5 and 1 mg/mouse induced a significant stimulation of renal ODC activity in a dose-dependent manner when it was given alone or in combination with testosterone. Using a sensitive increase in ODC activity in response to androgens as an end point, we did not detect an antiandrogenic effect of several antiandrogens, such as cyproterone acetate, spironolactone, p,p'DDE and vinclozolin. Also, none of these antiandrogens were able to change the basal level of renal ODC activity, with the exception of cyproterone acetate that at a dose of 0.1 mg/mouse stimulated ODC activity. The data obtained suggest that mouse renal ODC from CBA females is not strictly androgen-specific and cannot be used for estimation of antiandrogenic effects of compounds having an affinity to different types of receptors.     

Phenoloxidase activity in the hemolymph of the spiny lobster Panulirus argus

The prophenoloxidase activating system plays a major role in the defense mechanism of arthropods. In the present study, the phenoloxidase activity and its location in the hemolymph of the spiny lobster Panulirus argus is presented. Phenoloxidase activity was observed in the hemocyte lysate supernatant (HLS) and plasma after their incubation with trypsin. Higher amounts of trypsin were required to activate the HLS prophenoloxidase, due to the presence of a trypsin inhibitor in this fraction. Activation of prophenoloxidase was found when HLS was incubated with calcium, with an optimal pH between 7.5 and 8. This spontaneous activity is due to the prophenoloxidase activating enzyme, a serine proteinase that activates the prophenoloxidase once calcium ions were available. SDS was able to induce phenoloxidase activity in plasma and hemocyte fractions. Prophenoloxidase from HLS occurs as an aggregate of 300kDa. Electrophoretic studies combining SDS-PAGE and native PAGE indicate that different proteins produced the phenoloxidase activity found in HLS and plasma. Thus, as in most crustaceans, Panulirus argus contains a prophenoloxidase activating system in its hemocyte, comprising at least the prophenoloxidase activating enzyme and the prophenoloxidase. Finally, it is suggested that phenoloxidase activity found in plasma is produced by hemocyanin.     

Primary culture of hemocytes from the Caribbean spiny lobster, Panulirus argus, and their susceptibility to Panulirus argus Virus 1 (PaV1)

Primary cultures of hemocytes from the Caribbean spiny lobster Panulirus argus were developed for studies on the in vitro propagation of Panulirus argus Virus 1 (PaV1). A modified Leibovitz L-15 medium supported the best survival of hemocytes in in vitro primary cultures. However, degradation of the cultures occurred rapidly in the presence of granulocytes. A Percoll step gradient was used to separate hemocytes into three subpopulations enriched in hyalinocytes, semigranulocytes, and granulocytes, respectively. When cultured separately, hyalinocytes and semigranulocytes maintained higher viability ( approximately 80%) after 18 days incubation compared with granulocytes, which degraded over 2-3 days. Susceptibility of the cell types was investigated in challenge studies with PaV1. Hyalinocytes and semigranulocytes were susceptible to PaV1. Cytopathic effects (CPE) were observed as early as 12h post-inoculation, and as the infection progressed, CPE became more apparent, with cell debris and cellular exudates present in inoculated cultures. Cell lysis was noticeable within 24h of infection. The presence of virus within cells was further confirmed by in situ hybridization using a specific DNA probe. The probe gave a unique staining pattern to cells infected with PaV1 24-h post-inoculation. Cells in the control treatment were intact and negative to hybridization. This assay was further applied to the quantification of infectious virus in hemolymph using a 50% tissue culture infectious dose assay (TCID(50)) based on CPE. These tools will now allow the quantification of PaV1 using established culture-based methods.     

The Effect of Environmental Processes on the Isomeric Composition of DDT-related Compounds

The pesticide p,p’-DDT (1-chloro-4-[2,2,2-trichloro-1-(4-chlorophenyl)ethyl]benzene) has been used in agriculture, forestry, and vector control for decades. Technical DDT, the purity commonly used for insecticidal applications, contained p,p’-DDT as well as its constitution isomer o,p’-DDT (1-chloro-2-(2,2,2-trichloro-1-(4-chlorophenyl)ethyl]benzene) and several closely related compounds. To investigate the effect of environmental processes on the isomer-specific composition of DDT and its degradation products, we examined previously published and newly acquired quantitative data with regard to o,p’-/p,p’-DDX ratios in different environmental matrices. Isomeric shifts related to the depositional regime, transfer between and within compartments, and microbial transformation were clearly observed. Shifts of the positional isomer ratios potentially can serve as indictors to track the environmental fate of DDX. For an unambiguous assignment of these shifts to specific processes, further systematic studies and complementary lab experiments are necessary.     

黄斑篮子鱼和金钱鱼鳃的扫描电镜观察

对两种鲈形目鱼类黄斑篮子鱼(Siganus oramin)和金钱鱼(Scatophagus argus)的鳃结构进行扫描电镜观察。结果表明,黄斑篮子鱼和金钱鱼鳃的表面结构及微细结构与其他硬骨鱼类基本相似,鳃丝表面都具有规则或不规则分布的环形微嵴、沟、坑、孔等结构。黄斑篮子鱼的鳃片中部鳃丝表皮有大量凸起,而端部鳃丝表皮的凹凸程度明显较低,黄斑篮子鱼的鳃小片高度较金钱鱼鳃小片高。黄斑篮子鱼和金钱鱼鳃上皮的扁平上皮细胞、氯细胞和黏液细胞的形态结构及数量分布存在细微的差异。黄斑篮子鱼鳃片鳃丝的端部和中部表面有黏液细胞,金钱鱼鳃丝表面的黏液细胞很难观察到,与大多数淡水鱼类相似。黄斑篮子鱼鳃丝表面分布的氯细胞数量多于金钱鱼,这可能与两种鱼生活环境、生活习性的长期演变相关。     

Specializations and polyphagy of Plebejus argus (Lepidoptera: Lycaenidae) in North Wales

Abstract. 1. The polyphagous butterfly, Plebejus argus L., was found to have specialized requirements: eggs are laid along vegetation/bare ground margins, larvae specialize on tender meristematic and reproductive growth, larvae and pupae are tended by Lasius ants, and warm microclimates are favoured.
2. These requirements are met in early successional habitats within several bio topes. These habitats are locally distributed, as is P. argus.
3. Populations on limestone, heathland and mossland biotopes did not differ greatly in ecologies. Given a choice of foodplants in captivity, larvae from heathland and limestone did show slight differences in preference, selecting some of the hostplants they normally encounter in the wild. Regardless of biotope, all these larvae were plant taxa generalists (on three plant families), but specialists on plant parts.
4. The decline of P. argus in Britain is attributed to loss of biotopes and to declining traditional management of them. Fewer habitats are available, and the distances between them are greater than formerly. As some of these habitats are short lived, and P. argus rarely flies far, the butterfly's decline is a compound effect of reduced areas of habitat and inability to colonize them.     

Comparative population genetics and phylogeography of two lacertid lizards (Eremias argus and E. brenchleyi) from China

Eremias argus and Eremias brenchleyi are lacertid lizards that are sympatric throughout the distribution of E. brenchleyi. We sequenced partial mitochondrial DNA from cytochrome (cyt) b gene for 106 individuals of E. argus from nine localities, and for 45 individuals of E. brenchleyi from five localities, in central and northern parts of North China. We determined 53 cyt b haplotypes from the E. argus samples, and 27 cyt b haplotypes from the E. brenchleyi samples. Only E. brenchleyi had followed a stepping-stone model of dispersal. Partitioned Bayesian phylogenetic analysis reveals that E. argus and E. brenchleyi are reciprocally monophyletic, and the divergence time between the two species was dated to about 4.1±1.2 million years ago. Geographical structuring of haplotypes is more significant in E. brenchleyi than in E. argus. Haplotypes of E. brenchleyi could be divided into four groups by the Yellow River and Taihang Mountains. Within-population genetic diversity indices are correlated neither with latitude nor with longitude. We calculated significant among-population structure for both species (E. argus: Φ(ST)=0.608, P<0.001; E. brenchleyi: Φ(ST)=0.925, P<0.001). Eremias brenchleyi has four independent management units, while E. argus has a more homogeneous genetic structure across its range. Our data show that: (1) the pattern seen in North American and European species that southern populations have higher genetic diversity as consequence of post-glaciation dispersal is absent in the two Chinese lizards; (2) the Yellow River and Taihang Mountains may have acted as important barriers to gene flow only in E. brenchleyi; and (3) genetic structure differs between the two lizards that differ in habitat preference and dispersal ability.     

Dichlorodiphenyldichloroethylene potentiates the effect of protein kinase A pathway activators on progesterone synthesis in cultured porcine granulosa cells.

The insecticide dichlorodiphenyltrichloroethane (DDT) and its major metabolite p,p'-dichlorodiphenyldichloroethylene (DDE) have been implicated as endocrine-modulating chemicals. The DDT metabolite p, p'-DDE has been found contaminating human tissues and follicular fluid because of dietary exposure. We investigated the effects of DDE on progesterone synthesis in a stable porcine granulosa cell line, JC-410, and in primary cultures of porcine granulosa cells. Progesterone synthesis was not affected by 0.1-100 ng/ml DDE in the JC-410 cells. However, 10 ng/ml DDE increased 8-bromo-cAMP (8-Br-cAMP)-stimulated progesterone synthesis 0.4-fold (P < 0.05) over the levels observed with 1 mM 8-Br-cAMP alone. The effect of cholera toxin (CT) on progesterone synthesis was increased 0.7-fold (P < 0.05) by 10 ng/ml DDE over the value observed with 30 ng/ml CT alone. In primary cultures of porcine granulosa cells, 10 ng/ml DDE potentiated CT-stimulated progesterone synthesis 1.2-fold over the value observed with CT alone. In the JC-410 cells, 1 and 10 ng/ml DDE increased CT-stimulated cytochrome P450-cholesterol side-chain cleavage (P450(scc)) mRNA levels 0.3- and 0.4-fold, respectively, over the values obtained with CT alone. Neither basal nor CT-stimulated cAMP levels were changed by DDE. We conclude that DDE affects granulosa cell response to protein kinase A activators by altering the expression of the P450(scc) gene.     

Review of Panulirus argus virus 1--a decade after its discovery

In 2000, a pathogenic virus was discovered in juvenile Caribbean spiny lobsters Panulirus argus from the Florida Keys, U.S.A. Panulirus argus virus 1 (PaV1) is the first naturally occurring pathogenic virus reported from lobsters, and it profoundly affects their ecology and physiology. PaV1 is widespread in the Caribbean with infections reported in Florida (U.S.A.), St. Croix, St. Kitts, Yucatan (Mexico), Belize, and Cuba. It is most prevalent and nearly always lethal in the smallest juvenile lobsters, but this declines with increasing lobster size; adults harbor the virus, but do not present the characteristic signs of the disease. No other PaV1 hosts are known. The prevalence of PaV1 in juvenile lobsters from the Florida Keys has been stable since 1999, but has risen to nearly 11% in the eastern Yucatan since 2001. Heavily infected lobsters become sedentary, cease feeding, and die of metabolic exhaustion. Experimental routes of viral transmission include ingestion, contact, and for newly settled juveniles, free virus particles in seawater. Prior to infectiousness, healthy lobsters tend to avoid diseased lobsters and so infected juvenile lobsters mostly dwell alone, which appears to reduce disease transmission. However, avoidance of diseased individuals may result in increased shelter competition between healthy and diseased lobsters, and greater predation on infected lobsters. Little is known about PaV1 outside of Mexico and the USA, but it poses a potential threat to P. argus fisheries throughout the Caribbean.     

Vasodilatory and hypoglycaemic effects of two pyrano-isoflavone extractives from Eriosema kraussianum N. E. Br. [Fabaceae] rootstock in experimental rat models

Zulu traditional health practitioners have claimed that the roots of Eriosema kraussianum N. E. Br. (Fabaceae) and other Eriosema species (Zulu indigenous umbrella name of "uBangalala") are effective remedies for the treatment of erectile dysfunction (ED) and/or impotence. In order to scientifically appraise the significance and contribution of Eriosema kraussianum to its ethnomedical use as "uBangalala" and "VIAGRA substitute", the present study was undertaken to investigate the vasodilatory and hypoglycaemic properties of the two main bioactive chemical compounds obtained from E. kraussianum, in experimental rat models, using sildenafil citrate (VIAGRA) as the reference drug for comparison. The two E. kraussianum rootstock constituents (K1 and K2, 20-80 mg/kg p.o.) caused dose-dependent and significant (P<0.05-0.001) hypoglycaemia in rats. Relatively low to high concentrations of the plant's extracts (K1 and K2, 100-2000 microg/ml) always produced biphasic effects on rat isolated portal veins. K1- and K2-provoked responses of the isolated portal veins always consisted of concentration-related initial transient, but significant (P<0.05), contractions of the venous muscle preparations, followed by secondary, longer-lasting, highly significant (P<0.01-0.001) relaxations of the venous muscle strips. Sildenafil citrate (VIAGRA, 5-100 microg/ml) always produced concentration-related and highly significant relaxations of the rat isolated portal veins. Unlike K1 and K2 (20-80 mg/kg p.o.), however, sildenafil citrate (VIAGRA, 100 mg/kg p. o.) only caused slight and insignificant (P>0.05) reductions in the blood glucose levels of the experimental animals used. On the other hand, glibenclamide (10 mg/kg p.o.) induced highly significant (P<0.05-0.001), marked reductions in the blood glucose concentrations of the rats. The findings of this laboratory animal study indicate that the two hydro-ethanol extractives of E. kraussianum (K1 and K2) possess hypoglycaemic and secondary, vasorelaxant effects in the experimental paradigms used.