In vitro tests to establish LC(50) and discriminating concentrations for fipronil against Rhipicephalus (Boophilus) microplus (Acari: Ixodidae) and their standardization

Laboratory test was carried out on larvae and adults of the cattle tick, Rhipicephalus (Boophilus) microplus, to determine fipronil toxicity. Adult immersion test (AIT, N = 26), larval immersion test (LIT, N = 71) and larval packet test (LPT, N = 41) were standardized using susceptible strain (Mozo). Dose-response curves were compared with a fipronil resistant strain. Four variables were analyzed from AIT results: mortality, weight of eggs on day 7 and on day 14, index of fertility, and index of fecundity. For larval test, dose mortality curves were analyzed. In spite of the high LC(50) variability, all variables determined for AIT were appropriate to discriminate both strains. AIT and LIT had more sensitivity than LPT, with larger resistance factors. It was used two times LC(99.9) as discriminating doses (DCs) following FAO suggestion. For mortality by AIT, LIT and LPT the DCs were estimated: 4.98 ppm, 7.64 ppm and 2365.8 ppm, respectively, for Mozo strain. DCs mortality values estimated for resistant strain by AIT, LIT and LPT were: 6.96 x 10(5) ppm, 343.26 ppm and 5.7 x 10(3) ppm, respectively and their respective resistant factors were: 202.4, 5.36 and 1.52. Protocols for AIT, LIT and LPT have been presented in this paper. (C) 2009 Elsevier B.V. All rights reserved.

Inheritance and relative dominance, expressed as toxicity response and delayed development, of phosphine resistance in immature stages of Rhyzopertha dominica (F.) (Coleoptera: Bostrichidae)

Fumigation of stored grain with phosphine (PH 3) is used widely to control the lesser grain borer Rhyzopertha dominica. However, development of high level resistance to phosphine in this species threatens control. Effective resistance management relies on knowledge of the expression of resistance in relation to dosage at all life stages. Therefore, we determined the mode of inheritance of phosphine resistance and strength of the resistance phenotype at each developmental stage. We achieved this by comparing mortality and developmental delay between a strongly resistant strain (R-strain), a susceptible strain (S-strain) and their F 1 progenies. Resistance was a maternally inherited, semi-dominant trait in the egg stage but was inherited as an autosomal, incompletely recessive trait in larvae and pupae. The rank order of developmental tolerance in both the sensitive and resistant strains was eggs > pupae > larvae. Comparison of published values for the response of adult R. dominica relative to our results from immature stages reveals that the adult stage of the S-strain is more sensitive to phosphine than are larvae. This situation is reversed in the R-strain as the adult stage is much more resistant to phosphine than even the most tolerant immature stage. Phosphine resistance factors at LC 50 were eggs 400×, larvae 87× and pupae 181× with respect to reference susceptible strain (S-strain) adults indicating that tolerance conferred by a particular immature stage neither strongly nor reliably interacts with the genetic resistance element. Developmental delay relative to unfumigated control insects was observed in 93% of resistant pupae, 86% of resistant larvae and 41% of resistant eggs. Increased delay in development and the toxicity response to phosphine exposure were both incompletely recessive. We show that resistance to phosphine has pleiotropic effects and that the expression of these effects varies with genotype and throughout the life history of the insect. © 2012.

Acute toxicity of atrazine to Oreochromis niloticus fingerlings

Acute toxicity of atrazine to Oreochromis niloticus was undertaken to find the lethal concentration (LC) 50 of atrazine using fingerlings. Different concentrations were prepared in mg/L. There were six different concentrations with a control and each treatment was replicated three times. A total number of twenty-one aquaria were used. The highest concentration was 30mg/L. Ten test organisms were used in each aquarium. At 24, 48, 72 and 96 hours there were LC 50 of 15.6mg/L, 14mg/L, 11mg/L and 9.4, respectively. At 24, 48, 72 and 96 hours there were mean survivals of 49.0%, 34.3%, 28.6% and 28.1%, respectively

Toxicity of methyl amine on Catla catla (Ham) fingerlings

The toxicity of methyl amine was studied by finding out its LC 50 values for Catla catla fingerlings. On the basis of LC 50 values, the harmless concentration of methyl amine was found to be 12.8 ppm. This indicates that methyl amine is fairly toxic to C. catla fingerlings and needs care for its disposal in aquatic environment.

Toxicity of four therapeutic compounds to fry of Indian major carps

Toxicity of four therapeutic compounds: sodium chloride, potassium permanganate, potassium dichromate and acriflavine to fry of Catla calla, Labeo rohita and Cirrhinus mrigala was tested. Bioassays were conducted in 5 and 10 liters glass jars in the laboratory for periods of 24-48 hrs. at temperatures of 26° and 32°C. LC 50 values of the chemicals were in the ranges of 5500 to 7500 ppm, 37.5 to 48 ppm, 92.5 to 125 ppm and 47.5 to 80 ppm respectively. The results obtained are of value in evolving cheap control methods against some common fish diseases. Sodium chloride and acriflavine can be widely used for chemotherapy and potassium dichromate is preferable to potassium permanganate for prophylaxis.

Studies on the toxicological effects of Asulox-40 and Emisan-6 to eggs and early life history stages of Sarotherodon mossambicus

Toxicological effects of Asulox-40 and Emisan-6 to eggs and early life history stages of Sarotherodon mossambicus were reported. 80% of egg hatching occurred in the controls, 1 p.p.m and 5 p.p.m concentrations of Asulox-40. 10 p.p.m. and 50 p.p.m. concentrations of the same toxicants had 70% and 60% hatchings while in Emisan-6 in the same concentrations the hatching were 70% and! 40%. In 100 p.p.m. concentration of both toxicants 20% incomplete hatching occurred. In Emisan-6 Lc 50 and Lc 100 values were recorded at 32 hand 96h respectively in 10 p.p.m. concentrations. In Asulox-40 the same values were recorded in 24h and 40h respectively at 50 p.p.m. concentration. The fish activity during the experimental period showed initial hyper activity. It was established that the Emisan-6 is more harmful to S. mossambicus than Asulox-40. The harmless concentrations of these chemicals were 1.2 p.p.m. for Asulox-40 and 0.6 p.p.m. for Emisan-6.

Acute toxicity of unionized ammonia to milkfish (Chanos chanos Forsskal) fry

The study was conducted to determine the effects of varying concentrations of ammonia to milkfish fry. Two runs of static 96h bioassays were conducted to determine the median lethal concentration (LC 50) of unionized ammonia (NH₃) to milkfish fry. Test concentrations were based on exploratory 24h and 48h bioassays and were made in three replicates. Reagent grade ammonium chloride (NH₄Cl) was used to adjust the level of unionized ammonia. The 96h median lethal concentration, determined by the Reed Muench method was calculated at 28.029 ppm NH₃ 29.69 ppm. Even at high concentrations of unionized ammonia, most of the fry mortality occurred after 48 to 96 hours exposure. Severe gill damage occurs only at concentrations above 20 ppm, especially above the LC 50. The high LC 50 value obtain shows that milkfish fry has great tolerance to ammonia, that even fry with severely-damaged gills can still recover days after it is returned to favorable culture condition. The result suggest that observed mortalities of milkfish fry under culture conditions are not due to ammonia toxicity.

Cadmium toxicity to embryonic-larval development and survival in red sea bream Pagrus major

At 18 degrees C and 33 psu, 24 and 48 h LC50 values of cadmium (Cd) for red sea bream Pagrus major embryos were 9.8 and 6.6 mg l(-1), respectively, while 24,48, 72, and 96 h LC50 values for larvae were 18.9,16.2, 8.0, and 5.6 mg l(-1), respectively, indicating that embryos were more sensitive to Cd toxicity than larvae. Cd concentrations at >= 0.8 mg l(-1) led to low hatchability (0-90% in >= 0.8 mg l(-1) solutions vs. 97-100% in lower ones), delay in time to hatch, high mortality (38-100% vs. 1-10%), morphological abnormality (42-100% vs. 1-10%), reduced length (3.55-3.60 vs. 3.71-3.72 mm) in the embryos and larvae. They were Cd concentration dependent and potential biological significant endpoints for assessing the risk of Cd to aquatic organisms. Heart beat and yolk absorption of the larvae were significantly inhibited at some high concentrations but they were not as sensitive as other endpoints to Cd exposure. (C) 2008 Elsevier Inc. All rights reserved.

对虾暴发性病害的流行病学研究及养殖扇贝大规模死亡的病原病理分析

本文主要从病理病原的角度研究了中国对虾、栉孔扇贝和海湾扇贝大规模死亡的原因，同时，对中国对虾暴发性病害的流行病学特点及传播机制进行了探讨。中国对虾暴发性病害主要由对虾白斑综合症病毒（WSSV）引起。利用病虾的血淋巴，进行病毒的分离纯化，可以最大限度地降低对虾组织中有机物的干扰，提高WSSV的纯度，同时减少了提取步骤和处理时间，病毒粒子更完整、活力更高。带病毒的组织液经过TritonX-100处理后，可得到较完整，纯度较高的病毒核酸。对虾白斑综合症病毒在灭菌海水中经过15天（22 ℃）便失去感染力；WSSV对紫外线比较敏感，经过紫外线（15W，波长253.7nm，20cm距离）照射20min后，全部失活。在对虾病毒人工感染试验中，采用LC＿（50）法和紫外分光光度计法对感染病毒进行定量，其中，LC＿（50）能够较准确地反映病毒粒子的活力，而紫外分光光度计测得的病毒悬液浓度易受病毒悬液中杂质的干扰，误差相对较大。环境温度对对虾白斑病的暴发有重要影响，水温在9－13 ℃时，感染病毒的健康虾可以成为病毒的携带者，但不发病；18－22 ℃时，人工感染的健康虾无论采取投喂、浸泡还是注射的侵染方式，均发病死亡。WSSV对不同种类对虾具有不同的致病力，通过试验发现，在常见的对虾种类中，南美白对虾对WSSV的抵抗力大于中国对虾和鹰爪虾。本文利用PCR及电镜技术，对中国对虾染病亲虾的卵巢和各期卵及其刚孵化出的无节幼体进行了病毒检测，染病亲虾刚产的卵及未产出的卵均没有发现病毒存在，在卵巢的结缔组织中检测到病毒存在，刚孵化出的无节幼体PCR检测结果呈阳性，电镜下观测到有病毒样颗粒侵染无节幼体的上层表皮组织。在卵黄内及卵膜的内外层均未检测到病毒，没有发现病毒经卵内及卵膜传播的证据。病毒很可能是在无节幼体期进入对虾体内的。通过PCR及电镜检测发现，对虾养殖环境中的脊尾白虾、日本美人虾、野生螺类、小杂蟹体内均有WSSV存在，说明这些环境生物可能是对虾病毒的中间宿主或保存宿主，在虾病的暴发中起重要传播作用。通过对对虾的养殖海水进行不同级别的过滤发现，虾池中微型生物的数量对WSSV的传播也有一定的影响，在对虾养殖过程中，利用20cm绢网过滤海水，可有效降低WSSV的传播。通过组织病理分析和电镜观察，发现在濒死的栉孔扇贝和海湾扇贝体内有寄生原核生物－类立克次氏体（RLO），同时还发现少量的病毒粒子。病理学研究表明，类立克次氏体（RLO），造成栉孔扇贝组织细胞严重病变，可能是引起栉孔扇贝大规模死亡的重要原因。

Toxicity of lead, cadmium and mercury on embryogenesis, survival, growth and metamorphosis of Meretrix meretrix larvae

In order to assess the toxicity of heavy metals on the early development of Meretrix meretrix, the effects of mercury (Hg), cadmium (Cd) and lead (Pb) on embryogenesis, survival, growth and metamorphosis of larvae were investigated. The EC50 for embryogenesis was 5.4 mu g l(-1) for Hg, 1014 mu g l(-1) for Cd and 297 mu g l(-1) for Pb, respectively. The 96 h LC50 for D-shaped larvae was 14.0 mu g l(-1) for Hg, 68 mu g l(-1) for Cd and 353 mu g l(-1) for Pb, respectively. Growth was significantly retarded at 18.5 mu g l(-1) (0.1 mu M) for Hg, 104 mu g l(-1) (1 mu M) for Cd and 197 mu g l(-1) (1 mu M) for Pb, respectively. The EC50 for metamorphosis, similar to 48 h LC50, was higher than 96 h LC50. Our results indicate that the early development of M. meretrix is highly sensitive to heavy metals and can be used as a test organism for ecotoxicology bioassays in temperate and subtropical regions.

Application of rotifer Brachionus plicatilis in detecting the toxicity of harmful algae

The toxicity of seven major HAB (harmful algal bloom) species/strains, Prorocentrum donghaiense, Phaeocystis globosa, Prorocentrum micans, Alexandrium tamarense (AT-6, non-PSP producer), Alexandrium lusitanicum, Alexandrum tamarense (ATHK) and Heterosigma akashiwo were studied against rotifer Brachionus plicatilis under laboratory conditions. The results show that P. donghaiense, P. globosa, P. micans, A. tamarense (AT-6), or A. lusitanicum could maintain the individual survival and reproduction, as well as the population increase of the rotifer, but the individual reproduction would decrease when exposed to these five algae at higher densities for nine days; H. akashiwo could decrease the individual survival and reproduction, as well as population increase of the rotifer, which is similar to that of the starvation group, indicating that starvation might be its one lethal factor except for the algal toxins; A. tamarense (ATHK) has strong lethal effect on the rotifer with 48h LC50 at 800 cells/mL. The experiment on ingestion ability indicated by gut pigment change shows that P. donghaiense, P. globosa, P. micans, A. tamarense (AT-6) and A. lusitanicum can be taken by the rotifers as food, but A. tamarense (ATHK) or H. akashiwo can be ingested by the rotifers. The results indicate that all the indexes of individual survival and reproduction, population increase, gut pigment change of the rotifers are good and convenient to be used to reflect the toxicities of HAB species. Therefore, rotifer is suggested as one of the toxicity testing organisms in detecting the toxicity of harmful algae.

Insecticidal activity of proteinous venom from tentacle of jellyfish Rhopilema esculentum Kishinouye

Insecticidal activity of proteinous venom from tentacle of jellyfish Rhopilema esculentum Kishinouye was determined against three pest species, Stephanitis pyri Fabriciusa, Aphis medicaginis Koch, and Myzus persicae Sulzer. R. esculentum full proteinous venom had different insecticidal activity against S. pyri Fabriciusa, A. niedicaginis Koch, and M. persicae Sulzer. The 48 It LC50 values were 123.1, 581.6, and 716.3 mu g/mL, respectively. Of the three pests, R. esculentuin full proteinous venom had the most potent toxicity against S. pyri Fabriciusa, and the corrected mortality recorded at 48 It was 97.86%. So, S. pyri Fabriciusa could be a potential target pest of R. esculentum full proteinous venom. (c) 2005 Elsevier Ltd. All rights reserved.

Pruebas preliminares sobre toxicidad aguda del cobre en la Concha de Abanico (Argopecten purpuratus)

Se realizaron siete bioensayos estáticos con la Concha de Abanico (Argopecten purpuratus) con concentraciones de cobre que variaron de 0.007 a 0.74 ppm. El agua de mar de donde provinieron los individuos usados en el experimento tenía concentraciones de cobre entre 0.005-0.007 ppm. La bioacumulación inicial en los animales varó de 1.69 a 6.50 ppm. Como resultado preliminar se determinó que 0.13 ppm es la concentración letal media (LC 50) en 96 horas.

The influence of organophosphorus pesticide-methylparathion on protein, lipid metabolism and detoxifying enzymes in rohu (Labeo rohita)

Methylparathion (MP) is an organophosphorus insecticide used world wide in agriculture due to its high activity against a broad spectrum of insect pests. The aim of the study is to understand the effect of methylparathion on the lipid peroxidation, detoxifying and antioxidant enzymes namely catalase (CAT), glutathione peroxidase (GPx), superoxide dismutase (SOD), glutathione Stransferase (GST), total reduced glutathione (GSH), lipid peroxidation (LPO), acetylcholinesterase (AChE) and disease diagnostic marker enzymes in liver, sarcoplasmic (SP) and myofirbirllar (MF) proteins in muscles, lipids and histopathlogical changes in various organs of Labeo rohita of size 75 i 6g at lethal and sublethal level of exposure. The probit analysis showed that the lethal concentration (LC 50%) for 24, 48, 72 and 96h were 15.5mg/L, 12.3mg/L, 11.4mg/L and 10.2mg/L respectively which is much higher compared to the LC50 for juvenile fish. The LPO level and GST activity increased five folds and two folds respectively on exposure to methylparathion at 10.2 mg/L and the level of the enzymes increased, on sub lethal exposure beyond 0.25mg/L. AChE activity was inhibited by 74% at a concentration of 1.8mg/L and 90% at 5.4mg/L. The disease diagnostic marker enzymes AST, ALT, ALP and LDH increased by about 2, 3 ,3 and 2 folds respectively at pesticide concentration of 10.2mg/L when compared to control. On sub lethal exposure, however the enzymes did not show any significant changes up to 0.5mg/L. At a concentration of 10.2 mg/L, there was a three fold increase in myofibrillar proteins while the increase in sarcoplasmic protein was above 1.5 fold. On sub lethal exposure, significant alteration was noticed up to 30 days up to 1mg/L of methylparathion concentration. Further exposure up to 45 days increased sarcoplasmic proteins (upto 0.5mg/L). ln the case of myofibrillar proteins, noticeable changes were observed at 1mg/L concentration right from 15th day. The cholesterol content in brain tissues increased by about 27% at methylparathion concentration of 5.4 mglL. However at 0.25mg/L sub lethal concentration, no significant alteration was observed in enzyme activity, muscle proteins, lipids and histopathology of the tissues. The results suggest that methylparathion has the potential to induce oxidative stress in fish, and that liver, muscle and brains are more sensitive organs of Labeo rohita, with poor antioxidant potentials at higher concentrations of the pesticide. The various parameters studied in this investigation can also be used as biomarkers of methylparathion exposure.

Toxicological effects of Copper and Mercury on the fish Macrones gulio (Hamilton-Buchanan)

Asha M. R This thesis Entitled Toxicological effects of copper and mercury on the fish macerones gulio (hamiloton – buchanan).Chapter 1. In this chapter, a broad outline of heavy metal uptake, requirement of a suitable bio — monitoring organism, criteria for a standard test fish, and suitability of Macrones gulig for the toxicological study are given. Chapter 2. This chapter deals with the lethal toxicity bioassays to find the 96 hr LC 50 of copper and mercury for the fish Macrones gglig. The experimental results indicated that of the two metals tested, copper was more toxic than mercury.Chapter 3. The effect of copper and mercury on the haemoglobin, haematocrit, erythrocyte count, MCV, MCH and MCHC was studied.Chapter 4. The glycogen and protein contents of liver and muscle after exposure to copper and mercury were studied. There was a significant decrease of glycogen in the liver and muscle of metal treated fishes.Chapter 5. The histopathological changes of the tissues like liver, kidney and gill after exposure to copper and mercury were studied.