Developmental and reproductive effects in grass shrimp (Palaemon pugio) following acute larval exposure to a thin oil sheen and ultraviolet light.

Many early stages of estuarine species congregate at the surface or in the upper mixing layer making them prone to UV light exposure and oil sheens. Laboratory testing was used to assess UV-oil sheen interactions with grass shrimp (Palaemon pugio). Newly hatched grass shrimp larvae were exposed to a 1-µm thick oil sheen for 24 h with or without an 8-h pulse of UV light. Grass shrimp were then transferred to clean seawater and non-UV conditions to measure development, growth, and reproductive fitness. Minimal toxicity was observed after the initial exposure but larval development was significantly delayed in shrimp exposed to the UV enhanced sheen. After reaching sexual maturity, shrimp were paired to evaluate effects on reproduction. Shrimp initially exposed to the UV enhanced sheen as larvae had a significant reduction in fecundity compared to controls. This demonstrates the importance of examining interactions between UV light and oil since negative effects to aquatic organisms may be underestimated if based on standard laboratory fluorescent lighting. Acute exposures of early life stages to thin oil sheens and UV light may lead to long-term impacts to individuals and ultimately to grass shrimp populations.

Ultraviolet-B radiation induces transcriptional modulation of components associated with the extracellular matrix in embryos of decapod Macrobrachium olfersii.

The extracellular matrix (ECM) is a non-cellular and three-dimensional structure, constituted by a macromolecular dynamic network that involves the cells in all animal tissues, including embryonic ones. Several studies with vertebrates and cell cultures have reported deleterious effects of ultraviolet-B (UVB) radiation on the components associated with the ECM. However, studies focusing on the UVB radiation effects on ECM components of crustaceans during embryonic development are very scarce. Thus, the aim of this study was to identify the coding sequences of components associated with the ECM and to evaluate the effect of UVB radiation on embryos of the ecologically-important decapod Macrobrachium olfersii. To evaluate the modulation of these ECM components during embryonic development, the transcript levels of Col4α1, Itgß, Lamα, Mmp1 and Timp in M. olfersii embryos were analyzed at early developmental stages (E1, E3 and E4), intermediate developmental stage (E7) and late developmental stages (E10 and E14). In addition, embryos at E7, which correspond to a landmark of crustacean development, were analyzed after 12 h of UVB exposure to verify UVB effects on the ECM components. The ECM component sequences were similar to other decapods, suggesting conservation of these genes among crustaceans. The results showed modulations of the ECM components of M. olfersii embryos that reflect the need for each component in the cellular mechanisms, necessary for normal embryonic development. After UVB exposure, embryos showed opacity of embryonic tissues and it was found the overexpression of Col4α1, Itgß, Mmp1 and Timp transcript levels (1.82-, 1.52-, 2.34- and 6.27-fold, respectively). These impairments can compromise important events for normal embryonic development, such as growth of optic lobes, caudal papilla, ramification of appendages and differentiation of organic systems. The results presented here, together with the effects on morphology, cell proliferation, differentiation, and apoptosis demonstrated previously, strengthen the knowledge of the complex impacts of UVB radiation on freshwater embryos. Nevertheless, our results encourage further investigations focusing on the assessment of UVB effects on different organisms in order to better understand the myriad of UVB effects on ECM components.

Effect of cobalt-60 gamma radiation on total hemocyte content and biochemical parameters in Macrobrachium rosenbergii (De Man, 1879).

Purpose: The effect of low level cobalt-60 (60Co) gamma radiation on the freshwater prawn Macrobrachium rosenbergii was evaluated by observing their hemocyte counts and biochemical parameters. Materials and methods: Prawns were exposed to 3, 30, 300 and 3000 milligray (mGy) dose levels and their tissues of gills, hepatopancreas and muscle were analyzed. Results: The results showed that the number of hemocytes in the hemolymph and concentrations of protein and carbohydrate were significantly reduced in irradiated groups than compared to the control prawn. Increased aspartate transaminase (AST), alanine transaminase (ALT), lactate dehydrogenase (LDH), alkaline phosphatase (ALP), Acetyl choline esterase (AChE) in the irradiated groups reflects tissue damage. Conclusions: Hence, this study concludes that even low level of ionizing radiation (60Co gamma) can cause acute damages in gills, hepatopancreas and muscles in irradiated groups. Highlights 60Co exposures effect the THC and biochemical of prawn M. rosenbergii. Different dose levels such as 3, 30, 300 and 3000 mGy. Biochemical parameters serve as reliable indicators of physical status of organism. Self-regulating mechanisms might be the reason for preventing from the lethality. Suggested that nuclear industries should manage below 3 mGy.

Effect of UVB radiation exposure in the expression of genes and proteins related to apoptosis in freshwater prawn embryos.

Our previous studies showed that embryos of the freshwater prawn Macrobrachium olfersii exposed to ultraviolet B (UVB) radiation exhibited DNA damage, excessive ROS production, mitochondrial dysfunction and increased hsp70 expression, which are able, independently or together, to induce apoptosis. Thus, we attempted to elucidate some key apoptosis-related genes (ARG) and apoptosis-related proteins (ARP) and their expression during different stages of embryonic development, as well as to characterize the chronology of ARG expression and ARP contents after UVB radiation insult. We demonstrate that p53, Bax and Caspase3 genes are active in the embryonic cells at early embryonic developmental stages, and that the Bcl2 gene is active from the mid-embryonic stage. After UVB radiation exposure, we found an increase in ARP such as p53 and Bak after 3h of exposure. Moreover, an increase in ARG transcript levels for p53, Bax, Bcl2 and Caspase3 was observed at 6h after UVB exposure. Then, after 12h of UVB radiation exposure, an increase in Caspase3 gene expression and protein was observed, concomitantly with an increased number of apoptotic cells. Our data reveal that ARG and ARP are developmentally regulated in embryonic cells of M. olfersii and that UVB radiation causes apoptosis after 12h of exposure. Overall, we demonstrate that embryonic cells of M. olfersii are able to active the cell machinery against environmental changes, such as increased incidence of UVB radiation in aquatic ecosystems.

Developmental effects of exposure to ultraviolet B radiation on the freshwater prawn Macrobrachium olfersi: Mitochondria as a target of environmental UVB radiation.

In South America, increased UVB radiation has become an important environmental issue that is potentially threatening aquatic ecosystems. Considering that species exhibit different degrees of sensitivity to UVB radiation and that embryos are more sensitive than organisms at later life stages, the aim of this study was to characterize the effects of UVB radiation on subcellular compartments of embryos of the freshwater prawn Macrobrachium olfersi. This species lives and reproduces in clear and shallow waters, where UV radiation can fully penetrates. Embryos were irradiated with a UVB 6W lamp for 30min and examined after 1h, 12h, 24h and 48h of exposure. The irradiance of the UVB used simulates the UV radiation that embryos receive in the natural environment. The subcellular compartment most affected by the UVB radiation was the mitochondria, which exhibited a circular shape, a decrease in mitochondrial cristae, rupture of membranes and a morphology compatible with fission. These impairments were observed simultaneously with increased ROS production, just after 1h of UVB exposure. Thus, we investigated proteins related to mitochondrial fission (Drp-1) and fusion (Mfn-1), which are essential to cell maintenance. We found a significant increase in Drp-1 expression at all analyzed time-points and a significant decrease in Mfn-1 expression only after 24h of UVB exposure. Additionally, a decrease in embryonic cell viability was verified via the mitochondrial integrity assay. To conclude, we observed important mitochondrial dysfunctions against the environmental stress caused by UVB radiation. Moreover, the cellular responses found are critical and should not be disregarded, because they impact embryos that can potentially compromise the aquatic ecosystems.

Attenuation coefficients and absorbed gamma radiation energy of different varieties of potato, mango and prawn at different storage time and physiological condition.

Attenuation coefficients of different varieties of gamma irradiated potato (Kufri Chandramukhi, Kufri Jyoti, and Kufri Sindhuri), mango (Himsagar, Langra, Dashehri and Fazli) and prawn (Tiger prawn and Fresh water prawn) of different storage time and physiological stages were determined. After six months storage attenuation coefficient of Kufri Chandramukhi was decreased by 30.8% with decrease of density and moisture content. Decreasing trend of attenuation coefficient during storage was more prominent (almost 50%) in other two varieties of potato. On the other hand in all four varieties, unripe mango consisted of significantly less (p ≤ 0.05) attenuation coefficient (around 11-14%) than the ripe one due to changes in physiological properties and density. Different varieties of prawn had different attenuation coefficients due to subtle differences in their proximate composition. Due to having different attenuation coefficients, different food components, even different varieties of same food component absorbed different gamma radiation energy though exposed to same radiation dose.

Morphological and histological studies on freshwater prawn Macrobrachium rosenbergii (de man) irradiated with (60)Co gamma radiation.

This study was framed to investigate the (60)Co gamma radiation induced morphological and histological variations in freshwater prawn Macrobrachium rosenbergii. The LD50 value of (60)Co gamma irradiated M. rosenbergii observed (by probit analysis) at 30 Gy. Prawns were irradiated to four different dose levels (3 mGy, 30 mGy, 300 mGy and 3,000 mGy) using Theratron Phoenix TeleCobalt Unit [P-33] and one control group (without irradiation) maintained separately. Irradiated groups exhibited several morphological variations such as discoloration; damaged rostrum; opaque coloration in cephalothorax; black bands and dot formation in abdomen; deformed uropods and telson in tail regions when compared with control group. The Hepato Somatic Index reflected the severity of radiation on hepatopancreas. Histological variations in gills, hepatopancreas and muscles of irradiated groups were observed. In gills, structural changes such as swollen and fused lamellae, abnormal gill tips, hyperplasic, necrotic and clavate-globate lamellae were observed in gamma irradiated prawns. Accumulation of hemocytes in hemocoelic space, interstitial sinuses filled with abnormal infiltrated hemocytes, the tubular epithelium with ruptured basal laminae, abnormal and coagulated lumen, necrotic tubules, thickened basal laminae, tissue debris, necrotic hepatocytes were observed in irradiated prawn hepatopancreas. In muscle, shrinkage of muscular fiber and necrotic musculature were observed in irradiated prawns. These structural alterations of the organs it is felt could affect the vital physiological functions such as respiration, osmotic and ionic regulation in gills and muscles; absorption, storage and secretion of the hepatopancreas which in turn could adversely affect the growth and survival of freshwater prawn M. rosenbergii.

Effects of (60)Co gamma irradiation on behavior and gill histoarchitecture of giant fresh water prawn Macrobrachium rosenbergii (DE MAN).

Present study was designed to observe the effects of (60)Co gamma radiation in behavioral and histological changes in the gills of giant fresh water prawn Macrobrachium rosenbergii. The adult prawns were irradiated with four different dose levels (3mGy, 30mGy, 300mGy and 3000mGy) and the control group (without irradiation) was maintained separately. Behavioral changes like hyperactivity, loss of balance, reduced swimming rate, slower rate of food intake and convulsions were observed in higher dose levels of 300mGy and 3000mGy. The histological alterations such as accumulated haemocytes in haemocoelic spaces, abnormal gill tips, lifted lamellar epithelium, swollen and fused lamellae, hyperplasic, necrotic, clavate-globate and complete disorganization of lamellae were observed in (60)Co gamma irradiated prawns. Significantly more considerable histological alterations were observed in the highest dose level of 3000mGy, but no mortality was evidenced. This study serves as biomonitoring tool to assess the radiation pollution in the aquatic environment.

Effects of environmental and artificial UV-B radiation on freshwater prawn Macrobrachium olfersi embryos.

The recent decrease of the stratospheric ozone has resulted in an increase of ultraviolet-B (UV-B) radiation reaching the Earth's surface. In freshwater ecosystems with transparent water, UV-B rays easily penetrate and potentially cause harmful effects to organisms. In this study, embryos of the prawn Macrobrachium olfersi were used to evaluate the impact of UV-B rays in freshwater environments. We observed three groups of embryos: the first was to assess whether UV-B radiation produced morphological defects and/or biochemical impairments in the laboratory. The second was to check whether embryos with the same impairments as those observed in the laboratory were found in their environment, under natural solar radiation. The third group was the non-irradiated control. The embryos irradiated with 310 mW cm(-2) UV-B for 30 min showed morphological alterations similar to those observed in embryos from the environmental control group. The most important effects of the UV-B radiation observed in M. olfersi embryos were morphological (1.2% of the total number of embryos from the environment and 2.8% of the total number of irradiated embryos), pigmentation changes in the eyes (78.0% of the total number of embryos from the environment and 98.9% of the total number of irradiated embryos), and disruption of the chromatophores (46.9% of the total number of embryos from the environment and 95.5% of the total number of irradiated embryos). We also observed an increase in egg volume, which was accompanied by a significant increase in water content in UV-B irradiated groups when compared with aquaria control embryos. In addition, a significant decrease in the mitotic index in eggs exposed to UV-B radiation was detected (0.17 for the embryos from the aquaria control, 0.10 for the embryos of the environmental control, and 0.04 for the irradiated groups). The low levels of NPSH and high levels of TBARS indicated that UV-B rays directly compromised the antioxidant function of the embryonic cells, leading to oxidative stress. Our combined morphological and biochemical analyses revealed important effects induced by UV-B on M. olfersi embryos, and the results suggest that the recent changes in global conditions may have injurious effects, at least on the embryos of freshwater prawns.

Endocrine-mediated effects of UV-A irradiation on grass shrimp (Palaemonetes pugio) reproduction.

Although much is known regarding photoperiodic effects on crustacean egg production, the effects of ultraviolet (UV) light on reproduction has not been investigated. Likewise, little is known concerning the interaction between UV and xenobiotic exposure on crustacean reproductive cycles. In this study, male and female grass shrimp, Palaemonetes pugio, were exposed to sublethal concentrations of endosulfan (200 ng/l and 400 ng/l ES) under both white fluorescent (WF) and UV-A (315-400 nm) light conditions for 50 days in laboratory bioassays. Female endocrine (vitellogenin, ecdysteroids, and cholesterol), reproductive (percent gravid, clutch size), and embryo (days to hatch, hatching success, and hatching survival) responses were assessed. UV-exposure alone caused a significant (>4-fold) increase in total Palaemonetes pugio female egg production over the course of 50 days. Exposure to ES and UV significantly lowered the percentage of gravid females relative to UV controls, whereas ES-exposed shrimp under WF lighting did not exhibit these trends. Although higher vitellogenin concentrations and lower ecdysteroid titers were correlated with increased female egg production, cholesterol titers only exhibited a dose-dependent change when exposed to ES. Embryos from females exposed to UV had significantly lower ecdysteroid titers and shorter hatching times but there were no differences in embryo vitellogenin concentrations, hatching success, or hatching survival. These results indicate that UV-A exposure has a pronounced effect on grass shrimp (Palaemonetes pugio) reproduction and is likely mediated through 5-hydroxytrptamine (5-HT)-related neuroendocrine pathways. The implications for decapod aquaculture and evaluating chronic contaminant effects are discussed.