Sublethal Exposure to Phosphine Decreases Offspring Production in Strongly Phosphine Resistant Female Red Flour Beetles, Tribolium castaneum (Herbst)

The red flour beetle is a cosmopolitan pest of stored grain and stored grain products. The pest has developed resistance to phosphine, the primary chemical used for its control. The reproductive output of survivors from a phosphine treatment is an important element of resistance development but experimental data are lacking. We exposed mated resistant female beetles to 0.135 mg/L of phosphine for 48 h at 25°C. Following one week of recovery we provided two non-exposed males to half of the phosphine exposed females and to half of the non-exposed control females. Females that had been exposed produced significantly fewer offspring than non-exposed females. Females that remained isolated produced significantly fewer offspring than both exposed females with access to males and non-exposed controls (P<0.05). Some females were permanently damaged from exposure to phosphine and did not reproduce even when given access to males. We also examined the additional effects of starvation prior to phosphine exposure on offspring production. Non-exposed starved females experienced a small reduction in mean offspring production in the week following starvation, followed by a recovery in the second week. Females that were starved and exposed to phosphine demonstrated a very significant reduction in offspring production in the first week following exposure which remained significantly lower than that of starved non-exposed females (P<0.05). These results demonstrate a clear sublethal effect of phosphine acting on the female reproductive system and in some individuals this can lead to permanent reproductive damage. Pest population rebound after a fumigation may be slower than expected which may reduce the rate of phosphine resistance development. The results presented strongly suggest that phosphine resistance models should include sublethal effects. © 2012 Ridley et al.

Effects of sublethal dose of fipronil on neuron metabolic activity of africanized honeybees

Fipronil is a neurotoxic insecticide that inhibits the gamma-aminobutyric acid receptor and can affect gustative perception, olfactory learning, and motor activity of the honeybee Apis mellifera. This study determined the lethal dose (LD50) and the lethal concentration (LC50) for Africanized honeybee and evaluated the toxicity of a sublethal dose of fipronil on neuron metabolic activity by way of histochemical analysis using cytochrome oxidase detection in brains from worker bees of different ages. In addition, the present study investigated the recovery mechanism by discontinuing the oral exposure to fipronil. The results showed that mushroom bodies of aged Africanized honeybees are affected by fipronil, which causes changes in metabolism by increasing the respiratory activity of mitochondria. In antennal lobes, the sublethal dose of fipronil did not cause an increase in metabolic activity. The recovery experiments showed that discontinued exposure to a diet contaminated with fipronil did not lead to recovery of neural activity. Our results show that even at very low concentrations, fipronil is harmful to honeybees and can induce several types of injuries to honeybee physiology. © 2012 Springer Science+Business Media New York.

Brain morphophysiology of Africanized bee Apis mellifera exposed to sublethal doses of imidacloprid

Several synthetic substances are used in agricultural areas to combat insect pests; however, the indiscriminate use of these products may affect nontarget insects, such as bees. In Brazil, one of the most widely used insecticides is imidacloprid, which targets the nervous system of insects. Therefore, the aim of this study was to evaluate the effects of chronic exposure to sublethal doses of imidacloprid on the brain of the Africanized Apis mellifera. The organs of both control bees and bees exposed to insecticide were subjected to morphological, histochemical and immunocytochemical analysis after exposure to imidacloprid, respectively, for 1, 3, 5, 7, and 10 days. In mushroom bodies of bees exposed to imidacloprid concentrations of LD50/10 and in optic lobes of bees exposed to imidacloprid concentrations of LD 50/10, LD50/100, and LD50/50, we observed the presence of condensed cells. The Feulgen reaction revealed the presence of some cells with pyknotic nuclei, whereas Xylidine Ponceau stain revealed strongly stained cells. These characteristics can indicate the occurrence of cell death. Furthermore, cells in mushroom bodies of bees exposed to imidacloprid concentrations of LD50/10 appeared to be swollen. Cell death was confirmed by immunocytochemical technique. Therefore, it was concluded that sublethal doses of imidacloprid have cytotoxic effects on exposed bee brains and that optic lobes are more sensitive to the insecticide than other regions of the brain. © 2013 Springer Science+Business Media New York.

Zebrafish (Danio rerio) neuromast: promising biological endpoint linking developmental and toxicological studies

Aquatic toxicology is facing the challenge to assess the impact of complex mixtures of compounds on diverse biological endpoints. So far, ecotoxicology focuses mainly on apical endpoints such as growth, lethality and reproduction, but does not consider sublethal toxic effects that may indirectly cause ecological effects. One such sublethal effect is toxicant-induced impairment of neurosensory functions which will affect important behavioural traits of exposed organisms. Here, we critically review the mechanosensory lateral line (LL) system of zebrafish as a model to screen for chemical effects on neurosensory function of fish in particular and vertebrates in general. The LL system consists of so-called neuromasts, composed of centrally located sensory hair cells, and surrounding supporting cells. The function of neuromasts is the detection of water movements that is essential for the fish's ability to detect prey, to escape predator, to socially interact or to show rheotactic behaviour. Recent advances in the study of these organs provided researchers with a broad area of molecular tools for easy and rapid detection of neuromasts dysfunction and/or disturbed development. Further, genes involved in neuromasts differentiation have been identified using auditory/mechanosensory mutants and morphants. A number of environmental toxicants including metals and pharmaceuticals have been shown to affect neuromasts development and/or function. The use of the LL organ for toxicological studies offers the advantage to integrate the available profound knowledge on developmental biology of the neuromasts with the study of chemical toxicity. This combination may provide a powerful tool in environmental risk assessment.

The effect of sublethal concentration of DimilinR (TH6040) on the larval development of the mud-crab Rhithropanopeus harrisii (Gould)

Sublethal DimilinR (insecticide) concentration significantly decreased the percentage survival of larval R. harrisii . This decrease was marked at low salinities, when it may have become toxic. This was true for both 25 and 30 C. Duration of larval development did not seem to be affected by sublethal DimilinR concentrations even in extremely low or high salinities. High temperature, however, shortened the time of development. No anatomical abnormality was observed. These findings are only tentative as more replicates of the experiments are needed before definite conclusions can be reached. It will be desirable to work with more salinity-temperature combinations to get a complete picture of sublethal effects under different environmental conditions.

Effect of sublethal concentration of artificial seawater on oxygen uptake and opercular frequency in an Indian major carp, Labeo rohita (Ham.)

The present study reports the effect of artificial seawater on oxygen uptake and opercular frequency in an Indian major carp, Labeo rohita. Whereas a control fish of 7.34 g average body weight consumed 1.538 ml O sub(2.) hˉ¹, the 24h and 96h exposed fish of the same body weight consumed 1.07 4 and 0.897 ml O sub(2.) hˉ¹, respectively. The oxygen uptake per unit body weight under controlled condition was 0.219 ml. gˉ¹. hˉ¹, whereas in 24h and 96h exposed fish, it was 0.152 and 0.124 ml. gˉ¹, hˉ¹, respectively. The change in opercular movement in 24h exposed fish was 7.67% higher, whereas in 96h exposed fish, it was 22.43% higher as compared to the control one. All changes are highly significant (p<0.001).

Effect of sublethal doses of the insecticide imidacloprid on adaptive traits of Drosophila melanogaster: Response to treatment over and after 15 consecutive generations

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

The effect of organotin and copper sulfate on the late development and presettlement behavior of the hard clam Mercenaria mercenaria

The effect of organotin compounds and copper, commonly used as antifouling agent, were studied on Mercenaria mercernaria larvae. They were reared under usual hatchery conditions until they reached 190 um in diameter. The larvae were subjected to four compounds, tributylin chloride (TBT), monobutyltin chloride (MBT), trimethyltin chloride (TMT), cupric sulfate (CuSo4) plus control. Mortality was measured at 24, 48 h, and 96h. Behavioral and/or metamorphic changes were recorded in triplicate at 24-48 and 96 h. The appearance in swimming larvae of a functional foot was considered a sign of competence to set and was recorded as a "pediveliger". Swimming larvae were considered as larvae that have not yet reached their total development and they were recorded as "swimming". Larvae that did not show foot or swimming activity and were static but alive on the bottom were recorded as "bottom". TBT was found to completely inhibit swimming activity at sublethal concentrations throughout the period of observation. Copper and MBT inhibited swimming from 48 h, TMT did not inhibit swimming activity at any of the times recorded. The four compounds ranked in order of decreasing toxicity were TBT>TMT>CU>MBT.

Effect of diazinon 60 EC on Anabas testudineus, Channa punctatus and Barbodes gonionotus

Anabas testudineus, Channa punctatus and Barbodes gonionotus were exposed to 5.62, 6.25, 6.87, 7.50, 8.12 and 8.75 ppm; 1.13, 2.26, 3.39, 4.52, 5.65 and 6.78 ppm; and 2.00, 2.50, 3.00, 3.50, 4.00 and 4.50 ppm of Diazinon 60 EC, respectively. The median lethal concentration (LC50) values of Diazinon 60 EC on A. testudineus, C. punctatus and B. gonionotus were 6.55, 3.09 and 2.72 ppm for 96 hrs of exposure. The fish species showed several abnormal behaviors which included restlessness, arena movements, loss of equilibrium, increased opercular activities, strong spasm, paralysis and sudden quick movements during the exposure. For histopathological studies, A. testudineus, C. punctatus and B. gonionotus were exposed for 7 days to sublethal concentrations of 1.13 and 3.75 ppm; 1.13 and 2.26 ppm; and 1.13 and 2.26 ppm of Diazinon 60 EC, respectively. Hypertrophy, necrosis and pyknosis of hepatocytes, pyknosis and degenerative changes such as necrosis of tubular and haematopoietic cells of kidney were the major histopathological effects.

Effect of Neemta 2100 toxicity on acetylcholinesterase and serum glutamate oxaloacetate transaminase enzymes in serum of fish, Oreochromis mossambicus

Acetylcholinesterase and serum glutamate oxaloacetate transaminase enzymes have been used as marker monitoring the effect of neem seed based pesticide Neemta 2100 on the fish, Oreochromis mossambicus. Fishes exposed to sublethal concentrations of Neemta 2100 for acute periods of 24 and 48 hours were sacrificed to determine enzyme activities in serum affected due to toxicity. Laboratory studies of in vivo exposure of this pesticide showed synergistic inhibitory effect during acute period of toxicity. Acetylcholinesterase was noticed as 6.25 µm substrate hydrolyzed/mg protein/hour and serum glutamate oxaloacetate transaminase was noticed as 36.71 µm substrate hydrolyzed/mg protein/hour in control fish serum. Significant decrease in GOT level in Neemta 2100 treated fishes after short term exposure indicated its severe toxicity to fish.

Effect of copper, cadmium and Cu.Cd mixture on amino acid content in the poslarvae of penaeid shrimp, Penaeus monodon and P. penicillatus

This study document effects of short-term (96h) sublethal levels of copper, cadmium and their mixture on the amino acid composition of postlarvae of the penaeid shrimp, P.monodon and P.penicillatus . All experimental conditions were kept constant, temperature between 25-27•C and salinity 21-22 ppt. The estimated LD50 for Cu was 200 ug/L, for Cd 177.5 ug/L and for Cu.Cd mixture 250ug/L. In P. penicillatus at the same concentration of each metal, there was significant reduction in amino acid content, which was 8.01% higher than the control. Almost similar reduction in some amino acids was observed in P.monodon. At the maximum concentration of 400 ug/L, cadmium caused higher reduction in amino acid composition than did copper. Thus, amino acid composition may be regarded as a sensitive biochemical indicator of Cu and Cd toxicity because of the effect of these metals on protein synthesis, a signal of physiological stress in marine organisms subjected to heavy metal pollution.

Genotoxic effect of nickel chloride and zinc sulphate on fish Hypophthalmichthys molitrix

The present investigation is to assess the genotoxic potential of nickel chloride and zinc sulphate on gill cells of silver carp Hypophthalmichthys molitrix. Fishes were exposed in sublethal concentration of nickel chloride 5. 7 mg/1 and zinc sulphate 6.8 mg/1, and sampled at 10, 20 and 30 days. Nickel chloride and zinc sulphate treated fishes exhibited an apparent increase in the aberration frequency and a decrease in the mitotic index as compared to control. Acentric fragment, chromatid break, endoreduplication, chromatid gap, centromeric fusion, ploidy, sticky plate, dicentric chromosome, clumping and partial sticky plates were some of the abnormalities observed. The chromosomal aberrations in the treated fishes were significant compared to control.

Effect of long-term exposure to endosulfan on blood cell count in Channa gachua

In the present investigation, live specimens of Channa gachua were exposed to sublethal concentrations, i.e., 0.0017 and 0.00087 ppm of endosulfan for a period of 60 days. After the completion of 60 days, red and white blood corpuscles were counted from control as well as experimental fishes.