Bioaccumulation of TBT and Its Cellular Toxic Effects on the Freshwater Prawn Macrobrachium rosenbergii.

To test the toxic effects of tributyltin (TBT), Macrobrachium rosenbergii were exposed to three concentrations of TBT viz. 10 ng/L, 100 ng/L and 1000 ng/L for 90 days. The bioaccumulation of TBT level varied in hepatopancreas based upon dose dependent manner. Histopathological results revealed the reduction in basement membrane thickness, disruption of the hepatopancreatic tubules and abnormal lumen in hepatopancreas of TBT treated prawns. The ultrastructure of the control prawn showed normal architecture of cellular organelles with prominent nuclei in hepatocytes. On the other hand, many vacuoles, irregular arrangements of microvilli, swollen mitochondria, distorted rough endoplasmic reticulum cisternaes and abnormal nucleus were seen in the TBT treated group. Further, the biochemical and vitellogenin content were altered remarkably due to TBT exposure. It directly indicated that TBT had conspicuously inhibited the vitellogenesis. Therefore, it was inferred that the administration of TBT has considerably affected the hepatopancreatic functions in M. rosenbergii.

Detection of metal induced cytopathological alterations and DNA damage in the gills and hepatopancreas of green mussel Perna viridis from Ennore Estuary, Chennai, India.

This study report the impact of heavy metals on cytopathology and DNA damage in the gills and hepatopancreas of Perna viridis collected from Ennore estuary and the Kovalam coastal waters. Principal Component Analysis (PCA) showed significant differences among all variables at the scale of plots. The ultrastructural alterations such as lack of microvilli, distorted mitochondria, electron dense particles and the presence of large mucous droplets were common in the gill and hepatopancreatic cells of mussels from Ennore estuary. However, the gill and hepatopancreatic cells of P. viridis from Kovalam revealed normal compartmentalization of cells. The percentage of tail DNA in the mussels from Ennore estuary was recorded as 12.44 and 10.14% in the gills and hepatopancreas respectively. Overall, it has been demonstrated that the Comet and cytopathological assays are useful biomarkers to assess the level of pollution and it provide reliable information on ecotoxicology and genotoxicology of coastal waters.

Influence of short term exposure of TBT on the male reproductive activity in freshwater prawn Macrobrachium rosenbergii (De Man).

In the present study, the effect of tributyltin (TBT) on the histopathological and hormonal changes during spermatogenesis in freshwater prawn Macrobrachium rosenbergii was documented. Three experimental concentrations such as 10, 100 and 1,000 ng/L were selected and exposed to prawns for 45 days. After TBT exposure, the reproductive activities like sperm count and sperm length were decreased when compared with control. Further, abnormal structure of the seminiferous tubule, decrease in spermatozoa concentration, diminution of the seminiferous tubule membrane and the abundance of spermatocytes in the testis were noticed in treated prawns. Interestingly, radioimmunoassay clearly revealed the reduction of testosterone level in TBT exposed groups. Thus, TBT has considerably reduced the level of testosterone and caused the impairment of spermatogenesis in the freshwater male prawn M. rosenbergii.

Antifouling and toxic properties of the bioactive metabolites from the seagrasses Syringodium isoetifolium and Cymodocea serrulata.

The present study documents the antifouling and toxic properties of seagrasses Syringodium isoetifolium and Cymodocea serrulata. For that, the seagrasses S. isoetifolium and C. serrulata were extracted individually using organic solvents viz. dichloromethane, acetone and methanol. Amongst the extracts, the maximum antimicrofouling and antimacrofouling activities were exhibited by methanol extracts of both the seagrasses. The Minimal Inhibitory Concentration (MIC) of methanolic extracts of seagrasses was ranged from 1.0 to 10µg/ml against test biofilm bacteria and microalgal strains. Similarly, 100% fouling inhibition of limpet Patella vulgata was found at 6.0mg/ml of methanolic extracts of seagrasses. The mussel Perna indica showed 50% of byssal production and attachment inhibition at 21.51±2.03, 17.82±1.07µg/ml and the anticrustaecean activity for 50% mortality of Artemia salina was recorded at 732.14±9.21 and 394.16±5.16µg/ml respectively for methanolic extracts of S. isoetifolium and C. serrulata. The minimal inhibitory and higher lethal concentrations of active methanol extracts shows it׳s less toxic nature. Based on the prolific results, methanol extracts of S. isoetifolium and C. serrulata were subjected to purification using silica gel column and thin layer chromatography. Then the active compounds of the bioassay guided fractions were partially characterized using gas chromatography coupled with mass spectroscopy (GC-MS) and keyed out that fatty acids (C16 to C24) were the major components which responsible for the antifouling properties of the candidate seagrasses.

The application of histo-cytopathological biomarkers in the mud crab Scylla serrata (Forskal) to assess heavy metal toxicity in Pulicat Lake, Chennai.

The concentrations of heavy metals and their associated structural deformities in the gills, muscles and hepatopancreas of Scylla serrata from Pulicat Lake were determined and compared with crabs live along Kovalam coast. The concentrations of metals were high in the hepatopancreas and gills of crab from Pulicat Lake, whereas, low in crab from the Kovalam coast. Data were visualized using a principal component analysis (PCA). Significant differences were found for all variables at the plot scale; however, the overall variation was relatively low for muscle tissues in both stations. The structural deformities observed in the gills, hepatopancreas and muscle was due to metal toxicity, and the degree of damage was correlated with the elevated metal concentration. The results showed significant metal accumulation and histo-cytological lesions in the crabs from Pulicat Lake. The results suggest that these biomarkers are useful for assessing the impact of metal pollution in the coastal environments.

Ultrastructural changes during spermatogenesis, biochemical and hormonal evidences of testicular toxicity caused by TBT in freshwater prawn Macrobrachium rosenbergii (De Man, 1879).

The present investigation documents the impact of tributyltin (TBT) on the ultrastructural variation of spermatogenesis in freshwater prawn Macrobrachium rosenbergii. The environmentally realistic concentration of TBT can cause damages to the endocrine and reproductive physiology of crustaceans. In this context, three concentrations viz. 10, 100, and 1000 ng/L were selected and exposed to prawns for 90 days. The TBT exposed prawn exhibited decrease the reproductive activity as evidenced by sperm count and sperm length compared to control. Histopathological results revealed the retarded testicular development, abnormal structure of seminiferous tubule, decrease in the concentration of spermatozoa, diminution of seminiferous tubule membrane, abundance of spermatocytes and vacuolation in testis of treated prawns. Ultrastructural study also confirmed the impairment of spermatogenesis in treated prawns. Furthermore, radioimmunoassay (RIA) clearly documented the reduction of testosterone level in TBT exposed groups. Thus, TBT substantially reduced the level of male sex hormone as well as biochemical constituents which ultimately led to impairment of spermatogenesis in the freshwater male prawn M. rosenbergii.

Impact of TBT on the vitellogenesis and sex hormones in freshwater prawn Macrobrachium rosenbergii (De Man, 1879).

BACKGROUND: Tributyltin (TBT) is a ubiquitous persistent xenobiotic that can be found in freshwater, estuarine and marine ecosystem. TBT is a strong endocrine disrupting compound (EDC) that can cause toxic threat to aquatic organisms. Imposex, sexual deformities and endocrine dysfunctions are the causes of TBT to most of the aquatic organisms. Effect of TBT on the vitellogenesis and sex hormonal changes in Macrobrachium rosenbergii has never been reported. Hence, the present investigation was undertaken to find out the impact of TBT on histological changes in the different reproductive tissues, sex hormonal alterations and level of biomarkers like vitellogenin and vitellin in M. rosenbergii. RESULTS: The present investigation documents the possible impact of tributyltin (TBT) on the vitellogenesis in freshwater female prawn M. rosenbergii. TBT at 10 ng/l, 100 ng/l and 1000 ng/l concentrations were exposed individually to prawns for a period of three months. At higher concentration of 1000 ng/l, the ovarian development was arrested and ovary remained at spent stage. At lower concentration of TBT (10 ng/l), the development proceeded up to early vitellogenic stage. At intermediate concentration of 100 ng/l TBT, the ovary remained at pre vitellogenic stage and thereafter no development was noticed. Histological results indicated the normal ovarian development with vitellogenic oocytes, filled with yolk globules in control prawn. On the other hand, the TBT treated groups showed reduction in yolk globules, fusion of developing oocytes and abundance of immature oocytes. Immunofluorescence staining denoted the remarkable reduction in vitellin content in ovary of TBT treated prawn. Hence, TBT had conspicuously inhibited the vitellogenesis by causing hormonal imbalance in M. rosenbergii. CONCLUSION: TBT had notably inhibited the vitellogenesis due to hormonal imbalance. This endocrine dysfunction ultimately impaired the oogenesis in the freshwater female prawn M. rosenbergii.

Integrated use of histological and ultrastructural biomarkers in Mugil cephalus for assessing heavy metal pollution in Ennore estuary, Chennai.

Bioaccumulation of heavy metals and its associated histological perturbations were studied in various tissues of Mugil cephalus collected from Ennore estuary and compared with the fish collected from off-shore region. The concentration of copper, lead, zinc, cadmium, manganese and iron were quantified in gills, liver and muscle. The results showed marked differences between the two sites as well as significant variations within the tissues. The decreasing trend of metals in the tissues of fish sampled from both Ennore estuary and off-shore was in the order of Fe>Mn>Zn>Cu>Pb>Cd. Overall, the highest metal concentrations were found in the fish collected from Ennore estuary. The accumulation in the gills and liver of M. cephalus was found to be quite high in comparison with the muscle. These tissues were further investigated by light and electron microscopy and the results were compared with the reference site (off-shore region). The presence of large lipid droplets in liver and increase of mucous cells in gills were some of the most noticeable alterations observed and were related to heavy metal contaminants. It is concluded that histopathological and ultrastructural biomarkers provide reliable and discriminatory data to augment heavy metal pollution in Ennore estuary. Therefore, long-term monitoring is necessary to assess the eco-health of the Ennore estuarine environment by choosing bio-indicator species like M. cephalus, which provide accurate, reliable measurements of environmental quality.

Biomarkers of metal toxicity and histology of Perna viridis from Ennore estuary, Chennai, south east coast of India.

Distribution of heavy metals and its associated histological perturbations were studied in the soft tissues of Perna viridis collected from Ennore estuary and compared with the less polluted Kovalam coast. The concentration of copper, lead, zinc, cadmium, manganese and iron were quantified in gills, digestive gland and adductor muscle. The results showed marked differences between the two sites as well as significant variations within the tissues. Among the heavy metals analyzed, lead and cadmium recorded very low in the soft tissues of mussel. Heavy metal levels in tissues of mussel collected from Ennore estuary were in the order of gills>digestive gland>adductor muscle, while it was digestive gland>gills>adductor muscle in the mussel sampled from Kovalam coast. The decreasing trend of metals in the tissues of mussels sampled from both Ennore estuary and Kovalam coast was in the order of Fe>Mn>Zn>Cu>Pb>Cd. Overall, the highest metal concentrations were found in the mussel collected from Ennore estuary. The metal accumulation in the gills and digestive gland of Perna viridis was found to be quite high in comparison with the adductor muscle. These soft tissues were further investigated by light microscopy and the results were compared with the reference site (Kovalam coast). These results suggest that thickening of the digestive epithelium, hemocytic infiltration in the gills and myodegeneration in the muscle tissue are useful histological biomarkers for heavy metal induced stress, and demonstrate that precautions need to be taken in Ennore estuary in order to prevent heavy metal pollution that can occur in the future.

Effect of cadmium on the ovarian development in the freshwater prawn Macrobrachium rosenbergii (De Man).

In this investigation, effect of cadmium chloride (25 µg/l) on oogenesis of freshwater prawn, Macrobrachium rosenbergii was studied. In vivo experiments were performed with both intact and eyestalk ablated prawns. The intact, cadmium-exposed prawns exhibited decrease in Gonado Somatic Index (GSI) and ovarian development compared to controls. Whereas, ablated treated ovary showed reduction of yolk material and oocyte membrane thickness at the end of 15 days exposure. Interestingly, the control prawn showed normal cellular architecture of gills, hepatopancreas and ovary with mature oocytes. But, the gills of treated prawns showed lamellar hypertrophy, cuticular dystrophy and irregular arrangement of epithelial cells. Hepatopancreas showed reduction in both tubular diameter and basement membrane thickness. Conspicuously, ovary showed hypertrophied primary oocytes with more vacuoles in intact-treated group. Cadmium had increased gonad inhibiting hormone (GIH) secretion and decreased gonad stimulating hormone (GSH) release as evident with the retardation of gonadal maturation in the intact prawns.

Effect of tributyltin on the early embryonic development in the freshwater prawn Macrobrachium rosenbergii (De Man).

Organotin compounds particularly tributyltin (TBT) has been used as an antifouling biocides. In the present investigation, effects of TBT on the embryonic development, growth, hatchability and hatching success of eggs exposed to TBT were studied in both control and treated groups of the freshwater prawn (brooder) Macrobrachium rosenbergii. Three concentrations viz. 0.78ppm, 1.56ppm and 3.12ppm were selected for treatment. TBT at 3.12ppm concentration delayed the embryonic development and significantly reduced the hatchability of eggs as well. However, there were no significant changes obtained in the embryonic development and hatchability at the lower concentration (0.78ppm) of TBT. Whereas TBT at 1.56ppm affected the embryogenesis in toto. Interestingly, the control groups showed normal development such as elongation of body, formation of eyestalk and internal organs. On the other hand, the treated groups showed impaired embryonic development with reduced body growth. Interestingly, the embryo exposed to TBT showed absence of eyestalk and internal organs. Thus, TBT has significantly retarded the embryonic development in the freshwater prawn M. rosenbergii.