Combined effect of tributyltin and benzo[a] pyrene on the levels of sex hormone and vitellogenin in female Sebastiscus marmoratus

Tributyltin（TBT）, an organometal used as an antifouling biocide, has been reported to induce masculinization of fish. Benzo [a]pyrene （BaP）, a widespread carcinogenic polycyclic aromatic hydrocarbon, has been reported that its microsomal metabolites can produce an estrogenic response when tested in vitro. This study was therefore designed to examine the potential in vivo influence of TBT, BaP and their mixture on sex hormone levels in serum of Sebastiscus marmoratus, which were given 2 separate intraperitoneally （ip） injections（a single injection every 7 d） of TBT（0.5, 1, 5 and 10 mg/kg）, BaP（0.5, 1, 5 and 10 mg/kg）, or both in combination（0.5, 1, 5 and 10 mg/kg）; control fish received olive oil vehicle only. Six days after the 2nd injection, serum samples were collected and analyzed for sex hormone levels and alkali labile protein phosphorus （ALPP）, which is related to the yolk precursor protein vitellogenin. The pollutants at all doses significantly reduced serum testosterone, estradiol and ALPP content after 2 injections compared with the corresponding controls. The reduction of the estradiol levels should be response for the decrease of the vitellogenin levels. The results in the present study suggested that aromatase seems not the major target acted by TBT and BaP in fish. This study demonstrated that TBT or BaP exposure both inhibit the reproductive potential in female Sebastiscus marmoratus. Combined effect of TBT and BaP on the serum testosterone, estradiol and ALPP was not antagonism from the anticipation.     

Antioxidant responses to benzo[a]pyrene, tributyltin and their mixture in the spleen of Sebasticus marmoratus

It has been reported that there is an interaction between Benzo[a]pyrene （BaP）, a widespread carcinogenic polycyclic aromatic hydrocarbon, and tributyltin （TBT）, an organometal used as an antifouling biocide. This study was therefore designed to examine the potential in vivo influence of BaP, TBT and their mixture on splenic antioxidant defense systems of Sebastiscus marmoratus. The fish were exposed to water containing environmentally relevant concentrations of BaP, TBT and their mixture. Spleens were collected for biochemical analysis after exposure for 7, 25, 50 d and after recovery for 7, 20 d. Cotreatment with BaP and TBT for 7 d potentiated the induction of glutathione peroxidase （GPx） activity by BaP or TBT alone. The cotreatment for 25 and 50 d resulted in inhibition of GPx activity, which was similar to the effect of TBT. Splenic glutathione S-transferase （GST） activities were significantly elevated in S. marmoratus exposed to BaP starting from 7 d and remained high up to 25 d. However, no further activity change was found with prolonged exposure. Cotreatment of BaP and TBT primarily inhibited the GST activity, which was similar to the effect of TBT. Cotreatment with BaP and TBT for 25 or 50 d potentiated the depletion of GSH （glutathione） by BaP or TBT alone. MDA （malondialdehyde） contents in spleen of S. marmoratus were not significantly altered compared with the control during the test period. Spleen, as an immune organ, is sensitive to exposure of BaP or TBT. It should have an effective mechanism to counteract oxidative damage. Antioxidative defense systems in spleen of S. marmoratus should be considered as potential biomarkers. Short-term exposure of BaP or TBT could result in induction of antioxidant defense system. A significant decrease of these indices, such as GSH, GST, GPx might indicate more severe contamination.     

重金属Cd、Pb对褐菖鲉肝脏组织中HSPs基因表达的影响

为了解重金属污染对海洋鱼类热休克蛋白(HSPs)基因表达的影响,将褐菖鲉(Sebastiscus marmoratus)分别暴露于1.6、8、40、200、500μg·L~(-1)Cd、Pb溶液中,用环介导等温扩增技术(loop-mediated isothermal amplification,LAM P)定量检测褐菖鲉肝脏HSP60、HSP70、HSP90、HSC70 mRNA表达量。结果表明:Pb仅在40μg·L-1时显著抑制HSP60、HSP90、HSC70 mRNA表达量,8μg·L~(-1)时即可显著抑制HSP70 m RNA表达量,并在40μg·L~(-1)时达到最小值;Cd对HSP60、HSP90、HSC70的诱导不明显,但能显著诱导HSP70,并在500μg·L~(-1)时达到最大值。相比之下,褐菖鲉肝HSP70基因对重金属Cd、Pb污染较为敏感,有潜力成为监测海洋重金属污染的预警分子。     

Relation of hepatic EROD activity and cytochrome P4501A level in Sebastiscus marmoratus exposed to benzo[a]pyrene

This study was designed to investigate the in vivo effects of benzo[a]pyrene （BaP） on hepatic ethoxyresorufin-O-deethylase （EROD） activity and its correlation with cytochrome P4501A （CYP1A） protein levels in Sebastiscus marmoratus, which were exposed through a water column to BaP （10, 100, 1000 ng/L, respectively） or were treated with intraperitoneal injections of BaP （0.5, 1, 5, 10 mg/kg, respectively） every 7 d. The results showed that after 25 d of waterborne exposure to 1000 ng/L BaP, fish hepatic CYP1A levels and EROD activity were significantly induced. In contrast, EROD activity was not altered 7 d after second intraperitoneal injections, whereas, CYP1A protein levels were increased. Dose-dependent increase of biliary BaP metabolites demonstrated that the catalytic activity of CYP1A was induced by treatment with BaP. The lowest observable effect concentration with regard to biliary BaP metabolites （100 ng/L） was much lower than that with reference to EROD activity （1000 ng/L）. The results suggest that biliary polycyclic aromatic hydrocarbon （PAH） metabolites were shown to better reflect the contamination gradients of PAHs than EROD activity. It appeared to be necessary to measure CYP1A protein levels to complement the EROD activity in relevant toxicological assessments.     

Conservation of the Marbled Murrelet under the Northwest Forest Plan

Abstract:  The Marbled Murrelet ( Brachyramphus marmoratus ) was listed as threatened in 1992, primarily because of loss of its old-forest nesting habitat. Monitoring conducted over the first 10 years following implementation of the Northwest Forest Plan shows at-sea murrelet populations appear to be stationary, but recruitment is very low and demographic models project a 4–6% annual rate of decline. Monitoring of nesting habitat indicated there were about 1.6 million ha of higher-suitability nesting habitat on all lands at the start of the plan, about half of which occurred on federal lands. Most (88%) of higher-suitability habitat on federal lands was protected within reserves. Over the past 10 years, losses of habitat due primarily to fire have totaled about 2% on federal lands. Losses have been much greater (12%) on nonfederal lands, due primarily to timber harvest. Habitat is expected to accrue within reserves as younger forest matures and attains sufficient diameter to support nesting sites. At-sea estimates of population size are strongly and positively correlated with amounts of adjacent nesting habitat at a broad scale, supporting the idea that amounts of nesting habitat are a primary driver in wide-scale murrelet population distribution. Conditions at sea, however, such as temperature regimes, prey availability, and pollutants, continue to affect murrelet populations. The system of large reserves seems to have achieved the short-term objective of conserving much of the remaining nesting habitat on federal lands. These reserves are also likely to contribute to the long-term objective of creating large, contiguous blocks of nesting habitat. The plan has a primary role in conserving and restoring nesting habitat on federal land but will succeed in this role only if land allocations calling for such protection are in place for many decades.     

苯并(a)芘、三丁基锡及其混合物对褐菖鲉酚氧化酶活力的影响

以褐菖鲉为材料,每周一次腹腔注射浓度为0.5×10-6、1.5×10-6和10 ×10-6的苯并(a)芘(BaP)、三丁基锡(TBT)及两者的等比例混合物,观察鱼体肾脏、脾脏组织中酚氧化酶活力的变化.结果表明,实验浓度的这些持续有机污染物在短期内对褐菖鲉肾脏、脾脏中的酚氧化酶活力有一定程度的诱导激活作用,随着暴露时间的延长,酶活力下降,联合暴露可能会产生协同作用,毒性增强,诱发免疫毒性.     

Relation of hepatic EROD activity and CYP1A level in Sebastiscus marmoratus exposed to benzo[a]pyrene

This study was designed to investigate the in vivo effects of benzo[a]pyrene (BaP) on hepatic ethoxyresorufin-O-deethylase (EROD) activity and its correlation with cytochrome P450 1A (CYP1A) protein levels in Sebastiscus marmoratus, which were exposed through a water column to BaP (10, 100, 1000 ng/L, respectively) or were treated with intraperitoneal injections of BaP (0.5, 1, 5, 10 mg/kg, respectively) every 7 d. The results showed that after 25 d of waterborne exposure to 1000 ng/L BaP, fish hepatic CYP1A levels and EROD activity were significantly induced. In contrast, EROD activity was not altered 7 d after second ip injections, whereas, CYP1A protein levels were increased. Dose-dependent increase of biliary BaP metabolites demonstrated that the catalytic activity of CYP1A was induced by treatment with BaP. The lowest observable effect concentration with regard to biliary BaP metabolites (100 ng/L) was much lower than that with reference to EROD activity (1000 ng/L). The results suggest that biliary polycyclic aromatic hydrocarbon (PAH) metabolites were shown to better reflect the contamination gradients of PAHs than EROD activity. It appeared to be necessary to measure CYP1A protein levels to complement the EROD activity in relevant toxicological assessments.     

联苯菊酯暴露对雌性褐菖鲉卵巢发育的影响及其毒性机制

联苯菊酯是农业、园林等领域应用广泛的拟除虫菊酯类杀虫剂,然而有关联苯菊酯对鱼类生殖毒性的研究却很少。以常见的近海鱼类褐菖鲉为受试对象,探讨联苯菊酯对鱼类卵巢发育的影响及机制。褐菖鲉在以环境相关浓度(1、10、100 ng·L~(-1))的联苯菊酯暴露50 d后,各暴露组卵巢生殖细胞发育均受到显著抑制,卵巢17β-雌二醇与睾酮含量均出现下降,caspase-3活性呈剂量依赖性上升。相对定量PCR分析显示,卵巢雌激素受体(ERβ)mRNA水平下降。结果表明,联苯菊酯诱导细胞产生凋亡,导致雌、雄激素水平和雌激素受体表达下降,环境剂量作用下对褐菖鲉卵巢发育具有显著影响。     

褐菖鲇肝HSC70基因的环介导等温扩增技术的建立及在石油污染剂量效应研究中的应用

从褐菖鲉肝脏中克隆了热休克蛋白HSC70基因,利用环介导等温扩增技术(loop-mediated isothermal amplification,LAMP)建立HSC70基因的定量检测方法.为检测该方法的可行性,将褐菖鲉分别暴露于石油水溶性成分(water-soluble fraction,WSF) 20、60、180 μg·L-11d后,利用real-time PCR及LAMP技术同时测定褐菖鲉肝HSC70 mRNA表达量,两种方法测定结果基本一致,证实LAMP技术可用于褐菖鲉肝HSC70基因的定量.为更细致了解石油WSF影响褐菖鲉肝脏HSC70基因表达的剂量-效应关系,将褐菖鲉分别暴露于25、50、75、100、125、150、175 μg·L-1 WSF中,5d后采样,用LAMP技术定量检测HSC70 mRNA.结果表明,HSC70 mRNA表达量在50 μg· L-1浓度组即被显著诱导,在75μg·L-1浓度下达到最大值,这说明褐菖鲉肝HSC70基因对石油污染较敏感,有潜力作为海洋石油污染的生物标志物.     

Fragmentation effects on an endangered species across a gradient from the interior to edge of its range

Understanding how habitat fragmentation affects individual species is complicated by challenges associated with quantifying species-specific habitat and spatial variability in fragmentation effects within a species’ range. We aggregated a 29-year breeding survey data set for the endangered marbled murrelet (Brachyramphus marmoratus) from >42,000 forest sites throughout the Pacific Northwest (Oregon, Washington, and northern California) of the United States. We built a species distribution model (SDM) in which occupied sites were linked with Landsat imagery to quantify murrelet-specific habitat and then used occupancy models to test the hypotheses that fragmentation negatively affects murrelet breeding distribution and that these effects are amplified with distance from the marine foraging habitat toward the edge of the species’ nesting range. Murrelet habitat declined in the Pacific Northwest by 20% since 1988, whereas the proportion of habitat comprising edges increased by 17%, indicating increased fragmentation. Furthermore, fragmentation of murrelet habitat at landscape scales (within 2 km of survey stations) negatively affected occupancy of potential breeding sites, and these effects were amplified near the range edge. On the coast, the odds of occupancy decreased by 37% (95% confidence interval [CI] –54 to 12) for each 10% increase in edge habitat (i.e., fragmentation), but at the range edge (88 km inland) these odds decreased by 99% (95% CI 98 to 99). Conversely, odds of murrelet occupancy increased by 31% (95% CI 14 to 52) for each 10% increase in local edge habitat (within 100 m of survey stations). Avoidance of fragmentation at broad scales but use of locally fragmented habitat with reduced quality may help explain the lack of murrelet population recovery. Further, our results emphasize that fragmentation effects can be nuanced, scale dependent, and geographically variable. Awareness of these nuances is critical for developing landscape-level conservation strategies for species experiencing broad-scale habitat loss and fragmentation.