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Tributyltin (TBT) is recognized as a major environmental problem at a global scale. Haloalkaliphilic tributyltin (TBT)-degrading bacteria may be a key factor in the remediation of TBT polluted sites. In this work, three haloalkaliphilic bacteria strains were isolated from a TBT-contaminated site in the Mediterranean Sea. After analysis of the 16S rRNA gene sequences the isolates were identified as Sphingobium sp. HS1, Stenotrophomonas chelatiphaga HS2 and Rhizobium borbori HS5. The optimal growth conditions for biodegradation of TBT by the three strains were pH 9 and 7% (w/v) salt concentration. S. chelatiphaga HS2 was the most effective TBT degrader and has the ability to transform most TBT into dibutyltin and monobutyltin (DBT and MBT). A gene was amplified from strain HS2 and identified as TBTB-permease-like, that encodes an ArsB-permease. A reverse transcription polymerase chain reaction analysis in the HS2 strain confirmed that the TBTB-permease-like gene contributes to TBT resistance. The three novel haloalkaliphilic TBT degraders have never been reported previously.
Se considera a la tributiltina (TBT) como un problema medioambiental serio a escala global. Las bacterias haloalcalifílicas degradadoras de TBT pueden constituir un factor clave para remediar áreas contaminadas con dicho xenobiótico. En este estudio se aislaron 3 cepas de bacterias haloalcalifílicas procedentes de un sitio contaminado con TBT en el mar Mediterráneo. Tras analizar las secuencias del gen de 16S del ARNr, se identificaron los aislados como Sphingo-bium sp. HS1, Stenotrophomonas chelatiphaga HS2 y Rhizobium borbori HS5. Las condiciones de crecimiento óptimas para la biodegradación de TBT por parte de las 3 cepas fueron pH 9 y 7% (p/v) de concentración de sal. S. chelatiphaga HS2 fue el degradador de TBT más efectivo, con capacidad de transformar la mayor parte de ese compuesto en dibutiltina y monobutiltina (DBT y MBT). Se amplificó un gen de la cepa HS2, que fue identificado como tipo TBTB-permeasa, que codifica para una ArsB permeasa. Un análisis de la cepa HS2 por reacción en cadena de la polimerasa con transcriptasa inversa (RT PCR) confirmó que el gen TBTB-permeasa contribuye a la resistencia al TBT. Estos 3 nuevos degradadores haloalcalifílicos de TBT no habían sido reportados previamente.
Subject(s)
Bacteria/isolation & purification , Environmental Restoration and Remediation/methods , Biodegradation, Environmental , Mediterranean Sea/epidemiology , Polymerase Chain Reaction/methods , Reverse Transcription/genetics , /analysisABSTRACT
Objective To explore effects of parents exposure to TBT on blood routine of F1 generation mice. Methods 80 mice including 40 males and 40 females, were randomly divided into control groups (CK) , low dose groups (LTBT), middle dose groups (MTBT) and high dose groups (HTBT).They were given dose of TBT (0,0.2,2, 20μg/kg) every day.The experiment lasted 45 days.At 60 days, one female and one male of the same concentration were bred in the same cage according to 1∶1.At postnatal day 60, blood was collected for the determination of blood routine. Results Compared with control group, the number of red blood cells and hemoglobin of F1 generation male mice in LTBT and HTBT groups were significantly increased (P <0.01); Red blood cell volume, mean corpuscular hemoglobin (P <0.01), and the lymphocyte absolute value in F1 generation male LTBT were significantly reduced (P <0.05); HTBT of female mice were significantly increased about the number of red blood cells (P <0.01).A dose-dependent increase of the hemoglobin, red blood cells, and platelet count of F1 generation female experimental groups was observed.Conclusion Parental TBT exposure affects the F1 mice blood routine.There is the greatest influence on LTBT in F1 generation male mice and on HTBT in F1 generation female mice.
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Voluta musica is a dioecious marine gastropod endemic of the South Caribbean. Tributyltin (TBT) and copper (Cu) are potential inducers of imposex, an endocrine disorder by which females develop a penis and/or vas deferens. The goal of this work was to determine the imposex incidence in V. musica populations from Northeastern Península de Araya. For this, we selected three sites (Isla Caribe, Isla Lobos and Bajo Cuspe) and made monthly samplings of 15 snails in each site, during one year, and determined: (1) sizes; (2) sex and imposex incidence and (3) the Relative Penis Length Index (RPLI). We also performed histological analysis of the gonads, and measured TBT and Cu concentrations in sediments from the studied localities. Our results showed that the total number of sampled females affected by imposex was 24.5% at Isla Caribe, 12% at Isla Lobos, and none at Bajo Cuspe. In sediments, Cu was detected mostly in Isla Lobos. The female gonads with imposex did not show any development of male cells in any of the sampled sites. The higher percentage of females with imposex matched with the higher boat traffic locality, and higher TBT level (Isla Caribe). No esterilization was evident in this work, nevertheless, the presence of TBT and Cu in the sediments and females with imposex were considered as a potential threat to V. musica populations in this region. In Venezuela there is no control over this particular issue, possibly because of the lack of information and research in this topic, but certainly, this information will be useful in biodiversity conservation policies.
Voluta musica es un gasterópodo dioico endémico del Caribe sur. El TBT y el Cu, son potenciales causantes del imposex, fenómeno donde las hembras desarrollan un pene y/o vaso deferente. El objetivo fue determinar la incidencia de imposex en V. musica en el noreste de la Península de Araya. Se seleccionaron tres localidades y se captura-ron mensualmente 15 individuos durante un año para determinar: (1) talla de los individuos; (2) sexo y presencia de imposex; (3) índice Largo Relativo del Pene (RPLI). Se realizó histología de la gónada de los individuos. Se determinó TBT y Cu en el sedimento de cada localidad. En Isla Caribe, el 24.5% de las hembras presentó imposex, y se halló 3.9ngSn/g de TBT; en Isla Lobos, el 12% de las hembras desarrollaron imposex; en Bajo del Cuspe no se observó imposex. Se halló Cu en mayor concentración en Isla Lobos. Las gónadas femeninas con imposex no demos-traron masculinización. El mayor porcentaje de imposex coincide con la localidad de mayor tráfico de embarcacio-nes y con mayor nivel de TBT (Isla Caribe). No se eviden-ció esterilización, sin embargo la presencia de TBT, Cu e imposex son potenciales amenazas para las poblaciones de V. musica en la región. Hasta ahora, en Venezuela no se está tomando ninguna medida de control sobre este tema en particular, posiblemente por la escasez de información y orientación de las investigaciones hacia este tema, pero que sin duda se debería tomar en cuenta en las políticas para la conservación de la biodiversidad.
Subject(s)
Animals , Female , Male , Disorders of Sex Development/veterinary , Gastropoda/drug effects , Gonads/drug effects , Trialkyltin Compounds/toxicity , Water Pollutants, Chemical/toxicity , Disorders of Sex Development/chemically induced , Environmental Monitoring , Gonads/abnormalities , VenezuelaABSTRACT
OBJECTIVE: Tributyltin (TBT), an endocrine disrupting chemical, has been reported to decrease ovarian function by causing apoptosis in the ovary, but the mechanism is not fully understood. Therefore, we examined whether TBT increases the expression of adipogenesis-related genes in the ovary and the increased expression of these genes is associated with apoptosis induction. METHODS: Three-week-old Sprague-Dawley rats were orally administered TBT (1 or 10 mg/kg body weight) or sesame oil as a control for 7 days. The ovaries were obtained and weighed on day 8, and then they were fixed for terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) or frozen for RNA extraction. Using the total RNA of the ovaries, adipogenesis- and apoptosis-related genes were analyzed by real-time polymerase chain reaction (PCR). RESULTS: The ovarian weight was significantly decreased in rats administered 10 mg/kg TBT compared to that in control rats. As determined by the TUNEL assay, the number of apoptotic follicles in ovary was significantly increased in rats administered 10 mg/kg TBT. The real-time PCR results showed that the expression of adipogenesis-related genes such as PPARgamma, aP2, CD36, and PEPCK was increased after TBT administration. In addition, apoptosis-related genes such as TNFalpha and TNFR1 were expressed more in the TBT-administered rats compared with the control rats. CONCLUSION: The present study demonstrates that TBT induces the expression of adipogenesis- and apoptosis-related genes in the ovary leading to apoptosis in the ovarian follicles. These results suggest that the increased expression of adipogenesis-related genes in the ovary by TBT exposure might induce apoptosis resulting in a loss of ovarian function.
Subject(s)
Animals , Female , Rats , Adipogenesis , Apoptosis , DNA Nucleotidylexotransferase , In Situ Nick-End Labeling , Ovarian Follicle , Ovary , PPAR gamma , Rats, Sprague-Dawley , Real-Time Polymerase Chain Reaction , Receptors, Tumor Necrosis Factor, Type I , RNA , Sesame Oil , Trialkyltin Compounds , Tumor Necrosis Factor-alphaABSTRACT
OBJECTIVE: Tributyltin (TBT), an endocrine disrupting chemical, has been reported to decrease ovarian function by causing apoptosis in the ovary, but the mechanism is not fully understood. Therefore, we examined whether TBT increases the expression of adipogenesis-related genes in the ovary and the increased expression of these genes is associated with apoptosis induction. METHODS: Three-week-old Sprague-Dawley rats were orally administered TBT (1 or 10 mg/kg body weight) or sesame oil as a control for 7 days. The ovaries were obtained and weighed on day 8, and then they were fixed for terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) or frozen for RNA extraction. Using the total RNA of the ovaries, adipogenesis- and apoptosis-related genes were analyzed by real-time polymerase chain reaction (PCR). RESULTS: The ovarian weight was significantly decreased in rats administered 10 mg/kg TBT compared to that in control rats. As determined by the TUNEL assay, the number of apoptotic follicles in ovary was significantly increased in rats administered 10 mg/kg TBT. The real-time PCR results showed that the expression of adipogenesis-related genes such as PPARgamma, aP2, CD36, and PEPCK was increased after TBT administration. In addition, apoptosis-related genes such as TNFalpha and TNFR1 were expressed more in the TBT-administered rats compared with the control rats. CONCLUSION: The present study demonstrates that TBT induces the expression of adipogenesis- and apoptosis-related genes in the ovary leading to apoptosis in the ovarian follicles. These results suggest that the increased expression of adipogenesis-related genes in the ovary by TBT exposure might induce apoptosis resulting in a loss of ovarian function.
Subject(s)
Animals , Female , Rats , Adipogenesis , Apoptosis , DNA Nucleotidylexotransferase , In Situ Nick-End Labeling , Ovarian Follicle , Ovary , PPAR gamma , Rats, Sprague-Dawley , Real-Time Polymerase Chain Reaction , Receptors, Tumor Necrosis Factor, Type I , RNA , Sesame Oil , Trialkyltin Compounds , Tumor Necrosis Factor-alphaABSTRACT
Organotin compounds are typical environmental contaminants and suspected endocrine-disrupting substances, which cause irreversible sexual abnormality in female mollusks, called "imposex". However, little is known about the capability of triorganotin compounds, such as tributyltin and triphenyltin, to cause disorders in the sexual development and reproductive functions of mammals, including humans and rodents. Moreover, these compounds can act as potential competitive inhibitors of aromatase enzyme and other steroidogenic enzymes, affecting the reproductive capacity of male and female mammals. In this review, we discuss the cellular, biochemical, and molecular mechanisms by which triorganotin compounds induce adverse effects in the mammalian reproductive function.
Subject(s)
Animals , Female , Humans , Male , Genitalia/drug effects , Mammals/physiology , Organotin Compounds/toxicity , Trialkyltin Compounds/toxicity , Aromatase/drug effects , Endocrine System/drug effects , Reproduction/drug effectsABSTRACT
Objective To explore the effects of perinatal exposure to low dose tributyltin (TBT) on development and estrogen level of female offspring KM mice.Methods The CL healthy adult pregnant KM mice were randomly divided into 4 groups,6 in each group,they were given doses of TBT (100,10 and 1 ?g/kg) by gavage from days 6 of gestation to the end of lactation,1 time each day,at a volume of 1 ml/kg.On postnatal day (PND) 49,10 female offspring mice were randomly selected in each group and killed after weighed and the blood was collected.The uterus and ovary were weighed for account of viscera coefficient.The concentrations of estrogen in serum were measured with radioimmunoassay.Results Compared with the control group the time of eye opening was significantly delayed and the vagina opening was significantly ahead in treatment groups (P0.05).The weight of ovary and its coefficient increased significantly in 10 ?g/kg group (P
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Objective To explore the effects of gestation and lactation exposure to low dose tributyltin(TBT) on development of offspring of Kunming mice.Methods The healthy adult pregnant Kunming mice were randomly divided into 4 groups,6 in each group,they were given doses of TBT(100,10 and 1 ?g/kg) by gavage from days 6 of gestation(GD6)to the end of lactation.The number of pups and the number of male,female of each dams were determined,and the body weight of pups on PNDs 1,3,5,7,14,21 was weighed.The eye opening from PND12 and the testes descent from PND13 and the vagina opening from PND20 were examined.Results Compared with the control group,the gestational body weight gain of dams decreased in 100 ?g/kg group(P
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0.05).Compared with the control group,body weights gains from PND21-70 were decreased significantly in the 5 and 2.5 mg/kg group(P0.05).No abnormal structure of testis was observed in all exposure groups.Conclusion Exposure to TBTC during critical period for sex differentiation may decrease body gain and increase spermatogenesis and the activities of LDH and SDH.
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0.05). Compared with the control group,an obvious retarded development was observed from PND21-70 and the viscera coefficient of testis were increased significantly in 2.5 or 5.0 mg/(kg?d) group (P
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<p><b>OBJECTIVE</b>The aim of this study was to investigate the effect of tributyltin (TBT) compound onN-methyl-D: -aspartate (NMDA) receptors in the brains of preweanling mice.</p><p><b>METHODS</b>Pregnant ICR mice were exposed to TBT chloride at concentrations of 0, 15, and 50 ppm in water. Male offspring were sacrificed at 1, 2 and 3 weeks after birth. Mouse brain membranes were prepared from cerebral cortices, and the specific binding of [(3)H]MK-801 to an NMDA receptor was determined by radioligand binding assay.</p><p><b>RESULTS</b>The mean body weight of preweanling mice of the 50 ppm dose group decreased by 17-25% (p<0.01) at 1, 2 and 3 weeks of age, compared with that of preweanling mice of the corresponding control group. The [(3)H]MK-801 binding level significantly decreased (p<0.05) in the 15 ppm F1 group at 1 week and in the 15 ppm and 50 ppm F1 groups at 3 weeks of age, compared with that in the corresponding control F1 group.</p><p><b>CONCLUSIONS</b>The exposure to TBT via placenta and dam's milk seriously affected not only the growth of preweanling mice, but also the F1 cerebral NMDA receptors involved in memory and learning.</p>
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Objective: The aim of this study was to investigate the effect of tributyltin (TBT) compound on N-methyl-D-aspartate (NMDA) receptors in the brains of preweanling mice. Methods: Pregnant ICR mice were exposed to TBT chloride at concentrations of 0, 15, and 50 ppm in water. Male offspring were sacrificed at 1, 2 and 3 weeks after birth. Mouse brain membranes were prepared from cerebral cortices, and the specific binding of [3H]MK-801 to an NMDA receptor was determined by radioligand binding assay. Results: The mean body weight of preweanling mice of the 50 ppm dose group decreased by 17-25% (p<0.01) at 1, 2 and 3 weeks of age, compared with that of preweanling mice of the corresponding control group. The [3H]MK-801 binding level significantly decreased (p<0.05) in the 15 ppm F1 group at 1 week and in the 15 ppm and 50 ppm F1 groups at 3 weeks of age, compared with that in the corresponding control F1 group. Conclusions: The exposure to TBT via placenta and dam’s milk seriously affected not only the growth of preweanling mice, but also the F1 cerebral NMDA receptors involved in memory and learning.
Subject(s)
MiceABSTRACT
PURPOSE: The present study was performed to evaluate the effects of tributyltin acetate(TBTA) on mouse testes. The effects of TBTA on mammalian reproduction are not well known. MATERIALS AND METHODS: Three-week-old male mice(ICR strain) were orally administered TBTA at doses of 0 (control vehicle, CV), 25(T25), 50(T50), and 100 mg/kg(T100). Serum and intratesticular concentrations of testosterone and estradiol were determined by conventional radioimmunoassays. RT-PCR analysis was also performed. RESULTS: Transcriptional activity of 3-hydroxysteroid dehydrogenase (3beta-HSD), 17-hydroxysteroid dehydrogenase(17 beta-HSD) and cytochrome P450 17alpha-hydroxylase/C17,20 lyase(P450 (17 alpha)) were decreased by treatment. whereas mRNA levels of P450 aromatase were unaffected. In addition, TBTA significantly decreased serum testosterone levels in T100, while estradiol levels were not affected significantly. CONCLUSIONS: Administration of TBTA decreases testosterone level in testes, and this effect might be due to the alteration of mRNA levels of steroidogenic enzymes. Taken together, these findings suggest that TBTA, impairs testicular functions in a dose-dependent manner. The present results can be used as basic data in the study of TBTA action on gonads.
Subject(s)
Animals , Humans , Male , Mice , Aromatase , Cytochrome P-450 Enzyme System , Estradiol , Gonads , Oxidoreductases , Radioimmunoassay , Reproduction , RNA, Messenger , Testis , TestosteroneABSTRACT
<p><b>OBJECTIVE</b>Tributyltin (TBT) compounds have been widely used as antifouling agents for shipbottom paint. The immune system is a target of TBT intoxication. We evaluated the effects of TBT chloride in macrophages, which have critical roles in the immune system, using a murine macrophage lineage cell line, J774.1,in vitro.</p><p><b>METHODS</b>We examined tumor necrosis factor α (TNFα), interleukin-1β (IL-1β) andc-jun mRNA expression in J774.1 cells. The effects of TBT on the apoptosis of J774.1 cells were examined by determining the percentage of TUNEL-positive cells and caspase-3 activity.</p><p><b>RESULTS</b>The mean values of the viabilities of J774.1 cells exposed to TBT decreased dose-dependently. The relative mRNA expression of TNFα increased dose-dependently, however, that of IL-1β was not significantly different among the groups. The mean percentage of TUNEL-positive cells increased dose-dependently. Increases in the caspase-3 activities of J774.1 cells were observed in the groups exposed to higher concentrations of TBT. The mean value of relative mRNA expression of c-Jun transcription factor increased dose-dependently.</p><p><b>CONCLUSIONS</b>The increases in the percentage of TUNEL-positive cells and in caspase-3 activity suggested that exposure to TBT induces apoptosis of J774.1 cells. The increases in the mRNA expression of TNFα andc-jun by TBT may be related to apoptosis in macrophages.</p>
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Objective: Tributyltin (TBT) compounds have been widely used as antifouling agents for ship-bottom paint. The immune system is a target of TBT intoxication. We evaluated the effects of TBT chloride in macrophages, which have critical roles in the immune system, using a murine macrophage lineage cell line, J774.1, in vitro. Methods: We examined tumor necrosis factor α (TNF α), interleukin-1β (IL-1β) and c-jun mRNA expression in J774.1 cells. The effects of TBT on the apoptosis of J774.1 cells were examined by determining the percentage of TUNEL-positive cells and caspase-3 activity. Results: The mean values of the viabilities of J774.1 cells exposed to TBT decreased dose-dependently. The relative mRNA expression of TNFα increased dose-dependently, however, that of IL-1β was not significantly different among the groups. The mean percentage of TUNEL-positive cells increased dose-dependently. Increases in the caspase-3 activities of J774.1 cells were observed in the groups exposed to higher concentrations of TBT. The mean value of relative mRNA expression of c-Jun transcription factor increased dose-dependently. Conclusions: The increases in the percentage of TUNEL-positive cells and in caspase-3 activity suggested that exposure to TBT induces apoptosis of J774.1 cells. The increases in the mRNA expression of TNFα and c-jun by TBT may be related to apoptosis in macrophages.
Subject(s)
Tumor Necrosis Factor-alpha , RNA, Messenger , Apoptosis , In Situ Nick-End LabelingABSTRACT
Aim Organotin compounds are important organometallic chemicals with a variety of technical applications.Of these compounds,the trialkyltins,especially tributyltin,triphenyltin and trimethyltin,have been proved to have the distinct ability to induce tumor cell apoptosis and significant antitumor activity.They can induce tumor cell apoptosis through Fas receptor,mitochondrial,Ca 2+,MAPKs,p53,NF-?B and caspase/p38/ROS pathway.This paper reviewed recent progress in the studies on signal transduction in tumor cell apoptosis induced by trialkyltin compounds.
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The signaling pathways leading to cellular protection or cell death following exposure to heavy metals have not been fully clarified. Mitogen-activated protein kinases (MAPKs), i.e., extracellular signal-regulated protein kinase (ERK), c-Jun NH(2)-terminal kinase (JNK) and p38 MAPK transmit extracellular signals into the nucleus, and have been shown to participate in a diverse array of cellular functions such as cell growth, differentiation and apoptosis. Treatment with cadmium, inorganic mercury or tributyltin can activate ERK, JNK and p38 MAPK, and induces the expression of c-fos and c-jun genes prior to the development of apoptosis. However, the members of the MAPK family appear to be differentially activated depending on the heavy metal and the cell type exposed. Consequently, various cellular responses may be caused by the distinct pattern of MAPKs activation. MAPKs may be one of the important cellular signal transduction pathways affected by various environmental pollutants, including heavy metals.
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The signaling pathways leading to cellular protection or cell death following exposure to heavy metals have not been fully clarified. Mitogen-activated protein kinases (MAPKs), i.e., extracellular signal-regulated protein kinase (ERK), c-Jun NH2-terminal kinase (JNK) and p38 MAPK transmit extracellular signals into the nucleus, and have been shown to participate in a diverse array of cellular functions such as cell growth, differentiation and apoptosis. Treatment with cadmium, inorganic mercury or tributyltin can activate ERK, JNK and p38 MAPK, and induces the expression of c-fos and c-jun genes prior to the development of apoptosis. However, the members of the MAPK family appear to be differentially activated depending on the heavy metal and the cell type exposed. Consequently, various cellular responses may be caused by the distinct pattern of MAPKs activation. MAPKs may be one of the important cellular signal transduction pathways affected by various environmental pollutants, including heavy metals.
Subject(s)
Metals, Heavy , Cell Biology , Protein KinasesABSTRACT
Superoxide dismutase(SOD) was purified from Tegillarca granosa and the effects of tributyltin chloride(TBTCl) on the enzyme activity of the SOD were analyzed at different conditios. The results showed that the effects of TBTCl on SOD activity were more ohvious at room temperature than at four degrees centigrade. 58. 1 % of the enzyme activity was reserved after SOD was incubated with 200?g?mL-1 of TBTC1 for 72 hours at room temperature, while 90. 9% of the enzyme activity was reserved at four degrees centigrade. The enzyme activity of SOD was reduced with the increase of TBTC1 concentration and interaction time. TBTCl could reduce heat-resisting, acid-resisting and alkali-resisting abilities of SOD. With TBTCl concentration increase,SOD isozyme bands in the graph of polyacrylamide gel electrophoresis were getting weaker.
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Objective To explore the developmental toxicity of tributyltin chloride and the effects on sex hormone in female offspring rats through maternal gestation exposure. Methods Pregnant Wistar rats were randomly divided into four groups, 4 in each group. They were treated with TBTC by gavage at the dose of 0, 1, 2.5 and 5 mg/kg bw respectively from days 12-20 of gestation. 10 female offspring rats were randomly selected from each group and killed on postnatal day 70. The liver, kidney, uterus and ovary were weighed and the organ indexes were calculated. Pathological examination for liver, kidney, uterus and ovary were performed. Follicle stimulating hormone(FSH), luteotropic hormone(LH), testosterone(T) and estradiol(E2) in serum was determined by radioimmunity method. Results Increase of body weight in 2.5, 5 mg/kg bw groups significantly decreased(P