Acephate-induced shortening of developmental duration and early adult emergence in a nontarget insect Drosophila melanogaster

Similar to several environmental monitoring studies, the present study used Drosophila melanogaster as a model nontarget organism to explore the interfering effects of an organophosphate (OP) insecticide acephate on insect life cycle parameters. Acephate, a common OP, is readily available in nature from agricultural sources as an environmental contaminant. Along with target pests, nontarget fruit flies also suffer exposure to such environmental chemical. To evaluate the effects of such exposure, initially, acute LC₅₀ of acephate for third instar larvae was investigated and found to be between 14 and 16 μg/ml. This information yielded the following experimental concentrations (0.5, 1.5, 2.5, 3.5, 4.5, and 5.5 μg/ml) of test chemical for evaluation of effect, if any, on the insect model. Results showed that mean larval duration of insect significantly decreased on treatment with acephate, whereas the mean pupal duration remained unaffected. Interestingly the decreasing trend was seen to persist in case of mean adult emergence, where treated flies emerged significantly earlier in comparison to controls. Thus, the study demonstrated that acephate-induced shortening of developmental time and early emergence in Drosophila melanogaster.     

Kinetics of osmoregulation in the crab Carcinus maenas

When shore crabs Carcinus maenas are transferred from 11 to 38 S at 11°C, new constant levels of hemolymph freezing points and of concentrations of Na, K, Ca, and Mg in the hemolymph are accomplished within 24h. From a decrease in serum protein and in serum free amino acids and an increase in the relative amounts of individual essential free amino acids in the body fluids of whole crabs, a participation of proteolytic activities and a transport of amino acids from hemolymph to cell is deduced; the stationary concentration of total intracellular free amino acids increases up to a nearly two-fold value, compared with the concentration in crabs remaining in diluted sea water. Also, the low molecular neutral sugars increase in whole crabs after high salinity stress, reaching values of more than two-fold initial concentrations within a period of 10 days. This increase is fully accounted for by a 6.7-fold increase in trehalose levels. The processes of increasing the concentrations of low molecular organic material seem to be slower than those of establishing new osmotic pressures in the hemolymph. The oxygen consumption decreases by 30 to 45% to new constant values within 8 to 12 h after the salinity change from 11 to 38, reflecting similar kinetics as the establishment of new osmotic pressures in the hemolymph.     

Morphological and behavioral alterations induced by endocrine disrupters in amphibian tadpoles

Developmental toxicity effects of endocrine disrupter chemicals, acephate and cypermethrin were studied in Bufo melanostictus tadpoles. Thirty developing eggs of B. melanostictus were exposed to each concentration (0.01, 0.05, 0.1, 0.5, and 1?µg?L^?1) of acephate or cypermethrin in the laboratory (temperature: 23?±?1°C; photoperiod: 11.5–12.5?h). Eggs maintained in conditioned water alone served as controls. After hatching, larvae were fed on boiled spinach until the completion of metamorphosis. In control group, larvae that hatched on 3rd day were heavily pigmented, voracious feeders, and active swimmers; in these tadpoles, hind limb and forelimb-buds emerged on 16th and 24th day and metamorphosis was complete on 32nd day. Eggs exposed to acephate also hatched on 3rd day but larvae exhibited deformities such as, (i) tail distortions, (ii) laterally crooked trunk, (iii) decreased pigmentation, (iv) inactivity, (v) peeling of the skin, and (vi) delay in emergence of limbs and completion of metamorphosis. Cypermethrin-exposed eggs exhibited a delay (4–8 days) in hatching, there was no mortality, deformities in tail, trunk and head region, delay in the emergence of limbs, and completion of metamorphosis were evident. The demonstrated data indicate that these pesticides interfere with amphibian development when present in the aquatic system.     

Evaluation of hemopoietic responses in Labeo rohita Hamilton following acute copper toxicity

Fish are often exposed to highly contaminated water, especially in areas where the dilution rate of waste water is low. Environmental pollutants or other stress may affect one or more of the immunological functions in fish. Effective control of toxic substances is essential for the success of fish culture, which requires fish health monitoring using biomarkers. This study evaluated the effects of heavy metal copper (Cu) at environmentally relevant concentrations on the hematopoiesis in Labeo rohita. L. rohita fingerlings were exposed to different Cu concentrations for 6, 24, 48, or 72 hr. Following exposure, percentage blast cells was found to be increased after 6 and decreased after 24, 48, and 72 hr both at sublethal and LC₅₀ concentrations, whereas percentage mature erythrocytes decreased after 6 and increased after 24, 48, and 72 hr. Erythropoietic and leukopoietic efficiencies were also affected upon exposure. Erythropoietic and leucopoietic efficiencies increased markedly after 6, 24, and 48 hr but decreased after 72 hr in sublethal-treated fish. Erythropoietic efficiency decreased significantly after 6, 24 and 48 hr but increased after 72 hr in LC₅₀-treated fish. Leukopoietic efficiency decreased significantly after 6 hr exposure and increased after 24 and 48 hr but decreased after 72 hr. Flow cytometric studies of head kidney cell cycle phase distribution revealed cell death and DNA synthesis. The percentage cell death increased in fish exposed to sublethal concentrations and rose further at LC₅₀ during the earlier hours of exposure. Synthesis of DNA was increased at sublethal concentrations but was significantly reduced at LC₅₀ dose.     

Effects of the juvenile hormone mimic ZR-515 (Altosid®) on larval development of the mud-crab Rhithropanopeus harrisii in various salinities and cyclic temperatures

Effects of 0.01, 0.1 and 1.0 ppm methoprene (Altosid^®: ZR-515), a juvenile hormone (JH) mimic which shows high activity against some economically important insect pests, especially Diptera, were tested on larvae of the mud-crab Rhithropanopeus harrisii (Gould) (Brachyura: Xanthidae) from hatching to the first crab stage under optimum and stress conditions of a number of salinities and cyclic temperatures. There was a significant reduction in survival of zoeal larvae with increasing concentrations of methoprene in nearly all combinations of salinity and temperature. On the average there was 9% less survival in the 0.01 ppm concentration of methoprene than in the control, and in the 0.1 ppm concentration the survival was further reduced by another 16%. At 1.0 ppm methoprene no larvae survived beyond the first zoeal stage under optimum conditions or under stressful combinations of salinity and temperature. Except at 0.2 ppm in 27.5% S, survival of the megalopa was not significantly reduced in 0.01 or 0.1 ppm methoprene in any salinity or temperature, although the percentage of abnormal megalopa increased under stress conditions. The first zoeal stage was the most sensitive of the larval stages to methoprene as well as to salinity and temperature stress. The duration of zoeal development was significantly lengthened with an increase in concentration of methoprene under nearly all conditions of salinity and temperature. The JH mimic had, however, no significant effect on the duration of megalopa development. A significant synergism between methoprene, salinity and temperature was not observed. It can be concluded from the results that methoprene does not inhibit metamophosis of R. harrisii larvae at the 0.1 ppm level or lower. Reduction in survival of zoeal stages and increased duration of zoeal development with increasing concentrations of methoprene are presumably related to stress.     

Live algae as a vector candidate for hydrophobic polychlorinated biphenyls translocation to bivalve filter feeders for laboratory toxicity test

Live algae carrying hydrophobic xenobiotics can be an effective vector candidate for the chemical translocation to filter feeders in the laboratory toxicity test, but information on their application is lacking. Time-course uptake and elimination of polychlorinated biphenyls (PCBs) (0, 50, 100, and 500 ng g(-1) by two key algal foods, Isochrysis galbana and Tetraselmis suecica, were measured. Both of the algae achieved maximum concentration in an hourafter PCBs exposure regardless the chemical concentrations in our time-course measurements (0, 1, 5, 10, 24, 48 and 72 hrs). Once achieved the maximum concentration, the algae shortly exhibited elimination or eliminating tendency depending on the chemical concentrations. Algae exposed to the chemical for 1 and 24 hrs (hereafter 1 and 24 hr vectors, respectively) were then evaluated as a chemical translocation vector by feeding test to larval and spat Crassostrea gigas. In the feeding test the 24 hr vector, which contained lower chemicals than the 1-hr vector, appeared to be more damaging the early lives of the oyster. This was particularly significant for vectors of higher PCBs (p<0.05), probably due to algal reduction in food value by the prolonged chemical stress. These findings imply that 1 hr exposure is long enough for a generation of algal vector for laboratory toxicity test, minimizing data error resulted from reduction in food value by longer chemical stress.     

吡唑醚菌酯对不同阶段斑马鱼的毒性效应评价

为评价吡唑醚菌酯对鱼类不同生命阶段的毒性效应,以斑马鱼(Brachydanio rerio)为测试生物,测定了其对斑马鱼胚胎、仔鱼(孵化3 d和孵化12 d)和成鱼的急性毒性及对胚胎和仔鱼(孵化3 d)的发育毒性。结果表明,吡唑醚菌酯对斑马鱼胚胎、仔鱼(孵化3 d)、仔鱼(孵化12 d)和成鱼的96 h-LC50分别为0.0578(0.0540~0.0631)、0.0299(0.0282~0.0320)、0.0330(0.0310~0.0350)和0.0756(0.0692~0.0862)mg·L-1,其毒性大小依次为仔鱼(孵化3 d)仔鱼(孵化12 d)胚胎成鱼。与对照组相比,吡唑醚菌酯0.101 mg·L-1处理,可对斑马鱼胚胎24 h自主运动产生显著抑制作用;0.0609、0.0777和0.101 mg·L-1处理,可对斑马鱼胚胎48h心跳速率产生显著抑制作用;0.0609和0.0777 mg·L-1处理,可对斑马鱼胚胎96 h孵化仔鱼体长产生显著抑制作用。在供试浓度下吡唑醚菌酯暴露96 h后,均可对斑马鱼仔鱼(孵化3 d)体重和体长产生明显的抑制效应,对斑马鱼仔鱼(孵化3 d)心跳未产生明显影响。在高浓度下,吡唑醚菌酯可诱导斑马鱼胚胎产生畸形,主要包括心包水肿、卵黄囊水肿等;而对仔鱼(孵化3 d)未观察到明显的致畸效应。     

Influence of the polychlorinated biphenyl preparation Aroclor 1242® on color changes of the fiddler crab Uca pugilator

The melanin in the melanophores of specimens of the fiddler crab, Uca pugilator, exposed to 2, 4, and 8 ppm solutions of the polychlorinated biphenyl (PCB) preparation, Aroclor 1242^®, became less dispersed than in untreated specimens. This effect was dose-dependent. It was probably due to the PCB itself and not to polychlorinated dibenzofurans which are contaminants of commercial PCBs. The decreased melanin dispersion appeared to be related to a decrease in the rate of release of melanin-dispersing hormone from eyestalk neuroendocrine cells into the hemolymph. Eyestalks of crabs kept for 4 days in Aroclor 1242 contained 4 times as much melanin-dispersing hormone as did control crabs. When injected into isolated legs, did hemolymph from control crabs. Aroclor 1242 did not appear to have a direct effect on the pigment-dispersing mechanism of the melanophores.     

Adaptive behaviour of chironomid larvae (Chironomus riparius) in response to chemical stimuli from predators and resource density

Chemical-mediated effects of predatory fish on chironomid larvae behaviour have been ignored so far. Sediment-dwelling chironomid larvae inhabit protective burrows from which they extend their bodies only to feed on deposited detritus and microalgae from the surrounding sediment. Here, we performed factorial laboratory experiments to study whether fish-borne chemical cues (kairomones) are responsible for behavioural trait changes of chironomid larvae, and whether chironomid larvae are able to assess the densities of fish predators and food resources and the trade-off between them. We exposed naïve Chironomus riparius larvae to the chemical presence of zero, one, and ten predator fish (Rutilus rutilus) and offered two resource levels (low food, high food) for each treatment. Kairomones induced significant inherent behavioural trait changes in chironomid larvae. During the first 120 min after exposing chironomids to fish-conditioned water, we found a significant increase in digging activity with increasing predator density. After 3 days of exposure, the deepest chironomid burrows were found in treatments with the highest predator density. Chironomid larvae were significantly able to adjust their foraging behaviour to different predator densities and food concentrations and trade off between them; that is, when fish predators were more abundant or when more food resources were available, the foraging activities of larvae were significantly reduced. Our data suggest that chemically mediated trait changes (burrowing and foraging behaviour) may cascade through the littoral food web.     

Acute toxicity of ammonia to a freshwater teleost,Labeo bata larvae

Ammonia toxicity tests were performed with Labeo bata (bata) larvae of three different size groups. One hundred percent survival of larvae (500.0?±?4.0?mg) was recorded when exposed to ammonia concentrations of 1.0–13.56?mg?L^?1 at 96?h of exposure. Bata larvae exposed to ammonia concentrations of 15.8–25?mg?L^?1 showed 10–74% mortalities. The 96?h LC₅₀ value for 200 (±5), 250 (±2) and 500 (±4) mg bata larvae were 11.5, 16.8 and 22.5?mg?L^?1 un-ionised ammonia concentrations, respectively. When fish were exposed to different doses of ammonia, behavioural changes immediately occurred even at the lowest dose. At first, the fish became hyperexcitable, the skin darkened and they showed an increased ventilation frequency, fish behaviour became normal, 24?h after exposure. A 96?h LC₅₀ value of un-ionised ammonia showed direct relationship with the increasing size of bata larvae.     

Variation in UVB sensitivity of planula larvae of the coral Agaricia agaricites along a depth gradient

The effects of natural intensities of ultraviolet A (UVA, 320 to 400 nm) and B (UVB, 280 to 320 nm) radiation on planktonic planula larvae of the reef-building coral Agaricia agaricites (Linnaeus) were investigated through field experiments. Survival, chlorophyll concentrations, and solubilized protein concentrations were determined for larvae spawned from colonies at 3 and 24 m depth and subjected to one of three light regimes at 3, 10, or 24 m depth for 72 h: PAR (photosynthetically active radiation, 40- to 700 nm) only, PAR+UVA, or PAR+UVA+UVB. At 3 m depth, larvae in the PAR+UVA+UVB treatment showed lower survivorship than larvae exposed to either PAR alone or PAR+UVA. Within the PAR+UVA+UVB treatment at 3 m depth, larvae from colonies at 24 m depth suffered higher mortality than those from 3 m. Differences in survivorship between larvae originating from 3 and 24 m depth corresponded with tissue concentrations of UVB-protective mycosporine-like amino acids: larvae from 3 m had higher concentrations of mycosporine-glycine (_max=310nm) and palythine (_max=320nm) than those from 24 m depth. Chlorophyll concentrations were inversely correlated with PAR intensities, but variation in this parameter did not appear to be detrimental. These results show that sensitivity to high intensities of UVB radiation may affect survival of A. agaricites larvae in shallow reef-waters.     

Furanocoumarins and their detoxification in a tri-trophic interaction

The parsnip webworm, Depressaria pastinacella, specializes on wild parsnip, Pastinaca sativa, and several species of Heracleum, hostplants rich in toxic furanocoumarins. Rates of furanocoumarin metabolism in this species are among the highest known for any insect. Within its native range in Europe, webworms are heavily parasitized by the polyembryonic parasitoid wasp Copidosoma sosares. In this study, we determined whether these parasitoids are exposed to furanocoumarins in host hemolymph, whether they can metabolize furanocoumarins, and whether parasitism influences the ability of webworms to detoxify furanocoumarins. Hemolymph of webworms fed artificial diet containing 0.3 % fresh weight xanthotoxin, a furanocoumarin prevalent in wild parsnip hosts, contained trace amounts of this toxin; as well, hemolymph of webworms consuming P. sativa flowers and fruits contained trace amounts of six of seven furanocoumarins present in the hostplant. Thus, parasitoids likely encounter furanocoumarins in host hemolymph. Assays of xanthotoxin metabolism in C. sosares failed to show any ability to metabolize this compound. Parasitized webworms, collected from populations of Heracleum sphondylium in the Netherlands in 2004, were on average 55 % larger by weight than unparasitized individuals. This weight is inclusive of host and parasitoid masses. Absolute rates of detoxification (nmoles min⁻¹) of five different furanocoumarins were indistinguishable between parasitized and unparasitized ultimate instars, suggesting that the intrinsic rates of metabolism are fixed. Thus, although parasitized larvae are larger, detoxification rates are not commensurate with size; rates in parasitized larvae expressed per gram of larval mass were 25 % lower than in unparasitized larvae.     

Responses of echinoid larvae to food patches of different algal densities

High densities of larvae have been found in areas of high primary production, but it remains unclear whether this is the result of hydrodynamics or of larval aggregative behaviour in the presence of food. In this study, we examined changes in the vertical distribution and swimming patterns of four-armed larvae of the sea-urchin Echinometra lucunter (Linnaeus) around food patches of a range of microalgal densities. We reared larvae in the laboratory in a high or low concentration of either single (Isochrysis galbana) or mixed (I. galbana, Dunaliella tertiolecta, Thalassiosira weissflogii) microalgal species. In Plexiglas cylinders, we experimentally constructed haloclines in which the salinity of the bottom water-layer was 33‰ and that of the top water-layer was 24‰. In a thin layer in the middle of the halocline, we inserted a food patch that consisted of 0, 2500, 5000 or 10 000 T. weissflogii cells ml⁻¹. The presence of a food patch had a pronounced effect on the vertical distribution of larvae. This effect depended upon the algal density of the food patch and varied with dietary conditioning. The number of larvae that were above or within the patch decreased with increasing algal density, and was greater if larvae were reared in low-ration or single-species diets than in high-ration or mixed-species diets. Tracking of individual vertical swimming paths showed that within a few minutes, larvae swam into the patches of low algal density, and to positions just below the patches of the two higher algal densities, and remained there until the end of the experimental period. The greater number of algal cells in the digestive tracts of larvae from treatments with a food patch than in those without a patch confirmed that larvae were feeding on the microalgal cells of the patch. To our knowledge, this is the first study to experimentally show an aggregative behavioural response of invertebrate larvae to a food patch. Such a response may reduce the probability of food limitation and therefore enhance larval survival. Received: 14 February 1997 / Accepted: 24 September 1997     

Potential toxicity of flubendiamide in Drosophila melanogaster and associated structural alterations of its compound eye

Flubendiamide, a comparatively new insecticide, is used against lepidopteran insect pests and presumed safe for non-target Drosophila melanogaster (D. melanogaster). In this study, treatment concentrations (0.5, 1, 2 5, 10, and 20 μg/ml) of flubendiamide, lower than agricultural concentrations (rice 50μg/ml or cotton 100 μg/ml), were able to inhibit acetylcholinesterase activity in third instar larvae of D. melanogaster indicating a neurotoxic potential. In addition, larvae exposed to flubendiamide also manifested increased amounts of stress protein hsp70. The larvae expressing such stress response when allowed to emerge as adults displayed severe eye structure deformities found by scanning electron microscopy. These findings indicate a toxic potential for flubendiamide in D. melanogaster.     

Larval hemolymph feeding in the ant Leptanilla japonica by use of a specialized duct organ,the “larval hemolymph tap” (Hymenoptera: Formicidae)

Summary The larvae of the migratory Japanese ant Leptanilla japonica Baroni Urbani have a specialized duct organ on each side of the 4th abdominal segment. Behavioral and histological studies have shown that the adult ants are able to imbibe hemolymph directly from the larval body cavity through these organs, each of which is referred to here as a larval hemolymph tap. Laboratory observations further confirm that larval hemolymph feeding (LHF) is the sole source of nutrient for the queens. L. japonica is cyclical in its brood production. All larvae in a colony develop in concert, and when they are mature, the queen performs much active LHF on them. Nourished in this way, she achieves full physogastry within a period of only a few days and then lays a batch of 100–200 eggs. LHF thus facilitates synchronization of brood maturation and concentrated production of eggs by the queen. The larval hemolymph tap has probably evolved de novo in Leptanilla, in relation to cyclical brood production by species with small colony populations. Obviously, the function of this larval organ is non-selfish, contributing to overall colony-level functioning. The larvae of Leptanilla are thus properly qualified as a distinct class of caste in ant societies.     

Effect of salinity fluctuation on the osmotic pressure and Na+, Ca2+ and Mg2+ ion concentrations in the hemolymph of bivalve molluscs

Specimens of Chlamys opercularis, Modiolus modiolus, Mytilus edulis, Crassostrea gigas, Scrobicularia plana and Mya arenaria were exposed to both gradual (sinusoidal) and abrupt (square-wave) salinity fluctuations and measurements made of osmotic, Na⁺, Mg²⁺ and Ca²⁺ concentrations in the hemolymph and where applicable in the mantle fluid. In both sinusoidal and square-wave regimes fluctuating between 100 and 50% seawater (100%=ca. 32 S), the hemolymph Na⁺, Mg²⁺, Ca²⁺ and osmotic concentrations followed the concentrations of the external medium in Chlamys opercularis. The hemolymph and mantle fluid osmotic Na⁺, Mg²⁺ and Ca²⁺ concentrations of Modiolus modiolus, Mytilus edulis, Crassostrea gigas and S. plana followed those of the external medium as long as the molluscs' shell valves remained open. There were no changes in the ionic or osmotic concentrations of the hemolymph or mantle fluid of any of these species during periods of shell-valve closure. The hemolymph osmotic, Na⁺ and Mg²⁺ concentrations of wedged-open Modiolus modiolus, Mytilus edulis, C. gigas and S. plana followed those of the external medium. Hemolymph Ca²⁺ concentrations showed a damped response in C. gigas and Mytilus edulis. The hemolymph osmotic, Na⁺, Ca²⁺ and Mg²⁺ concentrations of Mya arenaria fluctuated in a similar manner to the external medium, but were damped. Wedged-open Mytilus edulis exposed to fluctuating salinity and supplied with a constant supply of 10 mM Ca²⁺ showed greater changes in hemolymph ionic and osmotic concentrations than M. edulis exposed to the same salinity fluctuation without a constant Ca²⁺ supply. Chlamys opercularis and Modiolus modiolus survived in a 50% seawater minimum sinusoidal salinity fluctuation for 10 days; wedged-open M. modiolus survived only 3 days. Burrowing had no effect on the osmotic, Na⁺, Mg²⁺ or Ca²⁺ concentrations of the hemolymph of Mya arenaria or S. plana exposed to fluctuating salinities. All of the species studied were shown to be osmoconformers.     

Carbofuran induced changes in breeding of a freshwater fish,Labeo rohita (Hamilton)

The effect of carbofuran, a carbamate pesticide, on the breeding performance of the freshwater fish Labeo rohita (Hamilton) has been investigated in the present study. Breeding of L. rohita was conducted after treatment with three sublethal concentrations, i.e. 0.06, 0.15 and 0.30 mg L^?1 of carbofuran for 96 h. The present investigation showed that the number of total eggs and total amount of eggs (litre per kg body weight) decreased significantly (p < 0.01) at all concentrations of carbofuran in comparison to controls, while the reduction in fertilization percentage at all concentrations of carbofuran was not significantly different from controls. The reduction in hatching percentage, the expected number of hatchlings, and the expected number of hatched larvae were significantly (p < 0.01) different between treated and control groups at all carbofuran concentrations. No significant differences for the 96 h survivability of hatched larvae were reported at all concentrations of carbofuran.     

The effect of queen-worker conflict on caste determination in the bumblebee Bombus terrestris

Endocrine analyses were used to investigate the well-known association between queen production and the onset of worker reproduction (termed the competition phase, CPh) in Bombus terrestris. Larvae that reached the age of 5 days before the CPh had a worker-like profile: low juvenile hormone (JH) biosynthesis rates and low JH hemolymph titers. In contrast, larvae that reached the age of 5 days during the CPh had a queen-like profile: high JH biosynthesis rates and high hemolymph JH levels. Larval fate could be manipulated by transplanting egg cells into host colonies with different social structures. There was a steep rise in JH production in larvae transplanted into colonies near or during the CPh. This indicates that during colony development, larvae switch from the ”worker developmental pathway” to the ”queen developmental pathway,” and that the switch is socially regulated. In small rearing groups, larvae reared with queens before the CPh developed into workers, whereas those reared with queens after the CPh developed into queens. Variation in worker type (naive or experienced) did not affect caste determination. Therefore, we hypothesize that queens produce a pheromone that directly inhibits queen differentiation by larvae. We also present two alternative scenarios that explain the timing of gyne production in B. terrestris, one based on ecological constraints and the other based on queen-worker competition. Received: 20 October 1999 / Received in revised form: 18 December 1999 / Accepted: 23 January 2000     

Antibacterial effects of Anodonta cygnea fluids on Escherichia coli and enterococci multi-drug-resistant strains: environmental implications

The persistence of bacteria in Anodonta cygnea largely depends on the hemolymph bactericidal activity against aquatic micro-organisms.

The aim of this study was to assess the in vitro bactericidal activity of whole hemolymph against multi-drug-resistant Escherichia coli and enterococci strains. Four mussels were injected with multi-resistant strains of E. coli in order to determine their in vivo bactericidal reactivity. In vitro experiments showed that the hemocytes viability decreased almost 70% in 4 h, the same happened in the control hemolymph. Enterococci tested in in vitro experiments were more susceptible than E. coli to whole hemolymph and hemocytes fractions. None of the enterococci bacteria was detected in the hemolymph fractions from 24 to 72 h, while E. coli was still detectable 72 h after inoculation both in the control and respective plasma fractions. The microbial inhibition on both hemolymph and cell pellets suggest that hemocytes may be the main responsibles for that process.

In vivo experiments showed that hemolymph inhibited the E. coli load injected and it was correlated with a high tendency for the increase in hemocytes counts, after 72 h. High adherence of E. coli and an intense pseudopods manifestation, mainly to the granulocytes, suggest a clear phagocytosis process.     

Biochemical and histopathological changes of Biomphalaria alexandrina snails exposed to bisphenol A

The effects of sub-lethal concentrations of bisphenol A on some biochemical parameters in Biomphalaria alexandrina snail's hemolymph and tissues as well as on histological changes of the hermaphrodite gland were evaluated. Adult snails were exposed to sub-lethal concentrations for four weeks and samples of snails were investigated after two and four weeks. Total protein and albumin levels in hemolymph, as well as lipid peroxidation and glutathione contents of snails’ tissues, were measured. After two weeks of exposure, total protein content and albumin levels decreased in all groups. Elevation of lipid peroxidation levels was correlated with decreasing glutathione content in the treated snails. Histological examination of the hermaphrodite gland revealed deformations in eggs and affected sperm production.