Doses Of Fipronil Research Articles

English

Pesticide use in the modern agriculture, animal husbandry and public health practices has increased enormously. Pesticides affect man, environment and wild life including birds. Pesticides including fungicides, herbicides and insecticides are the major class of chemicals deliberately released into the environment because of their indiscriminate use in various agricultural practices. Pesticides are present in air, soil and water which leave their residue in food chain causing deleterious effect on the health status of man and animals including poultry. After ingestion, they get into blood stream lodging themselves in various tissues causing adverse effects on health and production of livestock and poultry resulting in substantial economic losses. Fipronil was discovered and developed by Rhone-Poulenc between 1985- 1987 and placed in the market in 1993. It is the member of new and relatively small class of pesticides, the phenyl pyrazoles, which are principally chemicals with an herbicidal effect. According to data obtained from metabolic studies showed that there was potential for bioaccumulation of the Fipronil in fatty tissues, fish, fibre crops and food crops. Limited investigations have been carried on effect of fipronil in health of animals and man. Therefore present study was designed to study the effect of fipronil toxicity in white leghorn cockerels. Twenty white leg horn male chickens of one month old weighing about 150 to 200 g were procured and kept at Instructional Poultry Farm Nagla, Pantnagar. The birds in poultry shed in deep litter system under standard managemental conditions. They were randomly divided into four groups of 5 birds each. After one week of adaptation period, different doses of Fipronil were administered orally through feed at 1, 5 and 10 ppm levels to groups G1, G2 and G3 respectively for 100 days daily. The birds of group “C” served as control. After every 20 days blood was taken from each bird for study of different parameters. The experiment was conducted for a period of 100 days. All the hematological parameters viz. hemoglobin, packed cell volume, total erythrocyte count and differential leucocyte count were performed on the day of blood collection as per the standard method. Hematological parameters showed significant decrease leading to anemia and significant decrease in total leucocyte count leading to suppression in cell mediated immune response in different doses of fipronil treated birds in comparison to birds of control group.   Key words: Fipronil toxicity, white leghorn cockerels, haematological parameters.

Modification of the brain proteome of Africanized honeybees (Apis mellifera) exposed to a sub-lethal doses of the insecticide fipronil.

Fipronil is a phenylpyrazole insecticide that is widely used in Brazilian agriculture for pest control. Although honeybees are not targets of fipronil, studies indicate that this pesticide can be harmful to honeybees. To assess the effects of fipronil in the brain of Africanized Apis mellifera workers, this study focused on the toxico-proteome profiling of the brain of newly emerged and aged honeybee workers that were exposed to a sub-lethal dose (10pg fipronil per day. i.e. (1)/100 of LD50/bee/day during 5days) of the insecticide. Proteomic analysis identified 25 proteins that were differentially up-regulated or down-regulated when the fipronil-exposed and non-exposed groups were compared. These proteins are potentially related to pathogen susceptibility, neuronal chemical stress, neuronal protein misfolding, and occurrence of apoptosis, ischemia, visual impairment, damaged synapse formation, brain degeneration, memory and learning impairment. The exposure of honeybees to a very low dose of fipronil, even for a short period of time (5days), was sufficient to cause a series of important neuroproteomic changes in the brains of honeybees.

Prenatal exposure to a low fipronil dose disturbs maternal behavior and reflex development in rats

Fipronil (FPN) is a phenylpyrazole insecticide used in veterinary services and agriculture, and it is of considerable concern to public health. It inhibits the chloride channels associated with gamma-amino butyric acid (GABA) receptors in mammals and also inhibits the chloride channels associated with GABA and glutamate (Glu) receptors in insects. In this study, a commercial product containing fipronil was orally administered to pregnant Wistar rats at dose levels of 0.1, 1.0, or 10.0mg/kg/day from the sixth to twentieth day of gestation (n=10 pregnant rats/group). Its toxicity was evaluated based on maternal toxicity, reproductive quality, maternal behavior, and offspring physical as well as reflex development. All parameters observed in the observed offspring were assigned to one ink-marked couple in each litter (n=20 animals/group — 10 males and 10 females). The offspring couple represented the litter. Slight maternal toxicity presented during the second week of gestation for each fipronil dose and during the third gestational week at the highest dose due to lower chow intake. However, no effects were observed for gestational weight gain or gestation time, and the reproductive quality was not impaired, which suggests no adverse maternal effects from the doses during pregnancy. Moreover, the lowest fipronil dose compromised the active and reflexive maternal responses, but the highest dose induced a stereotyped active response without interfering in the reflexive reaction. For offspring development, no differences in physical growth parameters were observed between the groups. However, considering reflex development, our results showed that negative geotaxis reflex development was delayed in the offspring at the lowest fipronil dose, and palmar grasp was lost earlier at the lowest and intermediate fipronil doses. These results suggest that the alterations observed herein may be due to either the GABAergic system or endocrine disruption, considering that fipronil also acts as an endocrine disruptor.

Impact of fipronil on the mushroom bodies of the stingless beeScaptotrigona postica

Studies on stingless bees are scarce, and little is known about these insects, especially regarding the effects of contamination by neurotoxic insecticides, which can cause damage to important structures of the insect brain. This study evaluated the morphological changes in the intrinsic neurons of the protocerebral mushroom bodies (Kenyon cells) of the stingless bee Scaptotrigona postica after exposure to different doses of fipronil, using light microscopy and transmission electron microscopy. This region of the brain was selected for analysis because of its importance as a sensory integration centre. In both oral and topical treatments, Kenyon cells presented pyknotic profiles, suggesting cell death. Statistical analysis showed significant differences among doses and exposure times. Transmission electron microscopy revealed changes in the nucleus and cellular organelles. Depending on the dose, the characteristics observed suggested apoptotosis or necrosis. This study demonstrates the toxic effects of fipronil. An increase in the number of pyknotic profiles of Kenyon cells of mushroom bodies was observed even at the sublethal doses of 0.27 ng AI bee(-1) and 0.24 ng AI µL(-1) in the topical and oral treatments respectively. Also, differences in the number of pyknotic profiles were dose and time dependent.

In Vitro Genotoxicity of Fipronil Sister Chromatid Exchange, Cytokinesis Block Micronucleus Test, and Comet Assay

Fipronil (FP) is a phenylpyrazole pesticide developed by the transnational company Rhône-Poulenc Agro in 1987. Data on the genotoxicity and toxicity of FP are rather inadequate. In this study, we aimed to evaluate the potential genotoxic activity of FP using the single-cell microgel electrophoresis or comet assay, sister chromatid exchanges (SCEs), and micronuclei (MN) in human peripheral blood lymphocytes. In addition, the cytokinesis block proliferation index (CBPI) and proliferation index (PRI) were measured for cytotoxicity. In this study, three different doses of FP were used (0.7, 0.3, 0.1 μg/mL). Mitomycin C (2 μg/mL) and hydrogen peroxide were used as positive controls for SCE MN test systems, and comet assay, respectively. FP induced a statistically significant increase in the MN and SCE frequency and DNA damage in a dose-dependent manner in human peripheral blood lymphocytes (p<0.01, p<0.05, for 0.7 and 0.3 μg/mL, respectively) compared with a negative control. There is no significant difference between 0.1 μg/mL and the negative control for MN frequency, but there is significant difference between all the doses of FP and negative control for SCE frequency, mitotic index, CBPI, and PRI values (p<0.01). Using the alkaline comet assay, we showed that all the doses of the FP induced DNA damage in human peripheral blood lymphocytes in vitro (p<0.05).

Bioremediation of fipronil by a Bacillus firmus isolate from soil

Persistence of fipronil, a new molecule in extensive use against various insect pests is causing serious problems to the environment. Bacillus firmus was isolated by selective enrichment from soil samples collected from sugar fields with known history of pesticide usage and evaluated for metabolization of fipronil in clay loam soil. Soil samples in 50g aliquotes were fortified with fipronil @ 0.50–1.50mgkg−1 and inoculated with B. firmus cells (45×107CFUmL−1) and incubated at 25°C. Each sample in triplicates was drawn periodically up to 56d and residual fipronil contents analyzed by gas liquid chromatograph. Fipronil residues were not detected after 35d at lower doses of fipronil (@ 0.50, 0.75 and 1.00mgkg−1). However, at higher doses (@ 1.25 and 1.50mgkg−1) than this total metabolization of fipronil could be observed after 35 and 42d, respectively. Thus whereas, B. firmus proved its potential in efficient metabolization of fipronil, the period required for the same was dose dependent. Amongst various metabolites of fipronil degradation, fipronil sulfide was found to be the main metabolite followed by fipronil sulfone and fipronil amide. Though, desulfinyl metabolite earlier reported as one of the main metabolite of fipronil degradation, the existence of the same was not detected in any of the treatment.

Evaluation of Fipronil Oral Dosing to Cattle for Control of Adult and Larval Sand Flies Under Controlled Conditions

Visceral Leishmaniasis (VL) is a vector-borne disease endemic to the Indian subcontinent. The Phlebotomus genus of sand flies is the vector for VL in the Old World, with the vector on the Indian Subcontinent being Phlebotomus argentipes. Cattle are a commodity in this region and a frequent host source of P. argentipes bloodmeals. The purpose of this study was to determine the efficacy of a single oral dose fipronil against adult and larval P. argentipes. Ten Bos indicus cattle were used during the study in a controlled environment. The study was conducted in Bihar, India, and involved adult and larval bioassays using laboratory-reared P. argentipes. The results were positive in that they led to up to 100% mortality in both adult and larval sand flies over a 21-d period after a single dose of fipronil.

Cellular responses in the Malpighian tubules of Scaptotrigona postica (Latreille, 1807) exposed to low doses of fipronil and boric acid

Studies of sub-lethal effects of pesticide residues on stingless bees are scarce and morphological analysis of organs would add information to toxicological analysis in order to clarify the continuous exposure of Scaptotrigona postica to insecticides. The aim of this study was to evaluate the morphology and histochemistry of the Malpighian tubules (excretory organ) of S. postica exposed to fipronil or boric acid to detect cellular responses that indicate toxicity or adaptative mechanisms to stress induced by exposure of worker bees to low doses of these chemical compounds. Newly emerged bees were submitted to toxicological bioassays and morphological analyses by optical microscopy and Transmission Electron Microscopy, as well as histochemical methods, were performed to detect proteins and glycoconjugates. Additionally, immunohistochemical detection of DNA fragmentation and HSP70 (70-kDa Heat shock protein) were performed to detect cell death and stress response, respectively. Statistical analysis, for the bioassays conducted with ingestion of contaminated diet with boric acid at 0.75% (w/w) or with fipronil at 0.1μg/kg of food, showed that the survival of bees that ingested the contaminated diets were significantly different to the survival rate presented by the control group (P<0.0001). Although some characteristics indicative of initiation of cell death were observed, the cells remained metabolically active in the processes of excretion and inactivation of chemical compounds. The data from this study reinforce the importance of research on sublethal effects of low doses of pesticides on bees in an attempt to assess a possible realistic dose and evaluate the risk assessment of stingless bee S. postica foraging in the vicinity of cultivated fields and/or in green urban areas.

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.

Outbreak of Lice in Horses: Epidemiology, Diagnosis, and Treatment

The aim of this study is to report an outbreak of lice (Bovicola equi) in horses in southern Brazil, focusing on epidemiology, diagnosis, and treatment. Twenty-three horses in a stud farm presented with intense itching, crusted lesions, focal alopecia, and scaly skin. The outbreak occurred during the winter and probably due to the absence of sanitary barriers on the property. The diagnosis was carried out based on clinical signs associated with macroscopic and microscopic visualization of lice identified as B. equi. The treatment was performed with a single dose of Fipronil (Topline® Merial Brasil, Campinas, São Paulo, Brazil) spray (0.5%), eliminating immature and adult stages of lice. The clinical signs caused by the lice in horses disappeared few days after treatment.

Cytotoxicity of fipronil on mice liver cells

In the present study, mice livers were examined following exposure to different doses of fipronil (15, 25, and 50 mg/kg). Histological and histochemical techniques were used to determine the cytotoxic potential of this compound and to assess the damage it caused to livers. Mice were divided into four groups: control group and groups I, II, and III were exposed to 15, 25, and 50 mg/kg fipronil, respectively. Our findings revealed cytological, morphohistological, and histochemical alterations in liver cells of animals from groups I, II, and III compared to group control animals. These changes included Kupffer-cell proliferation, hepatocyte hypertrophy, accumulation and distribution of proteins, polysaccharides, lipids, and vacuoles in the cytoplasm of hepatocytes, and congestion of blood vessels. These phenotypes mainly characterize the following: (a) autophagic processes, (b) steatosis, and (c) cell death by necrosis, which demonstrate the damage caused by fipronil on nontarget organisms in artificial conditions.

Fipronil (active ingredient of acaricide frontline®) acting on the mice thyroid

Fipronil, active ingredient of Frontline® acaricide, is a phenyl-pyrazolic derivative, and its efficacy in the elimination of several plagues, even in lower concentrations, has already been demonstrated; however, its effect on nontarget organisms has not been thoroughly explained. In this sense, the present study had the objective to evaluate the effects of different doses of fipronil in mice and determine its cytotoxic potential and possible compromises on nontarget organisms in artificial conditions. The results showed that animals exposed to fipronil presented significant alterations in the thyroidean tissue with evident follicular disorganization, decreased in the size of most follicles culminating with the rupture of smaller follicles and still with posterior fusion in a big and irregular "follicle" filled with colloid. The present study also revealed that the action of fipronil not only causes disorganization in the thyroidean tissue but alsoalters the chemical composition of the colloid itself (prehormone), modifying its proteic basis, which was confirmed by histochemical test for protein detection, making it clear that, in thyroids exposed to fipronil, the colloid was weakly positive or even negative for the presence of proteins in its constitution.

Exposure to Sublethal Doses of Fipronil and Thiacloprid Highly Increases Mortality of Honeybees Previously Infected by Nosema ceranae

BackgroundThe honeybee, Apis mellifera, is undergoing a worldwide decline whose origin is still in debate. Studies performed for twenty years suggest that this decline may involve both infectious diseases and exposure to pesticides. Joint action of pathogens and chemicals are known to threaten several organisms but the combined effects of these stressors were poorly investigated in honeybees. Our study was designed to explore the effect of Nosema ceranae infection on honeybee sensitivity to sublethal doses of the insecticides fipronil and thiacloprid.Methodology/FindingFive days after their emergence, honeybees were divided in 6 experimental groups: (i) uninfected controls, (ii) infected with N. ceranae, (iii) uninfected and exposed to fipronil, (iv) uninfected and exposed to thiacloprid, (v) infected with N. ceranae and exposed 10 days post-infection (p.i.) to fipronil, and (vi) infected with N. ceranae and exposed 10 days p.i. to thiacloprid. Honeybee mortality and insecticide consumption were analyzed daily and the intestinal spore content was evaluated 20 days after infection. A significant increase in honeybee mortality was observed when N. ceranae-infected honeybees were exposed to sublethal doses of insecticides. Surprisingly, exposures to fipronil and thiacloprid had opposite effects on microsporidian spore production. Analysis of the honeybee detoxification system 10 days p.i. showed that N. ceranae infection induced an increase in glutathione-S-transferase activity in midgut and fat body but not in 7-ethoxycoumarin-O-deethylase activity.Conclusions/SignificanceAfter exposure to sublethal doses of fipronil or thiacloprid a higher mortality was observed in N. ceranae-infected honeybees than in uninfected ones. The synergistic effect of N. ceranae and insecticide on honeybee mortality, however, did not appear strongly linked to a decrease of the insect detoxification system. These data support the hypothesis that the combination of the increasing prevalence of N. ceranae with high pesticide content in beehives may contribute to colony depopulation.

Selective control of Rhipicephalus (Boophilus) microplus in fipronil-treated cattle raised on natural pastures in Lages, State of Santa Catarina, Brazil

An examination of a selective control of Rhipicephalus (Boophilus) microplus and consequent cost reduction was carried out in two areas of native pasture in the municipality of Lages, State of Santa Catarina, Southern Brazil, from May 2007 to April 2009. Forty cattle were divided into two groups of 20 animals each (conventional and selective control groups). At 14-day intervals female ticks larger than 4.5 mm found on the right flank of cattle bodies were counted, and these results multiplied by two. Fipronil 1 mg.kg(-1) pour on was then administered for tick control. In the conventional group all the cattle were treated when tick count averaged more than 40 female ticks per animal while in the selective group only animals parasitized by more than 40 ticks were treated with acaricide. Every 28 days the cattle's weight was checked for cost-effective analysis. The conventional group required an additional 20 doses of fipronil compared with the selective one. There was no statistically significant differences regarding weight gain in both groups.

Fipronil toxicity in northern bobwhite quail Colinus virginianus: Reduced feeding behaviour and sulfone metabolite formation

Fipronil is a phenyl pyrazole insecticide registered for agricultural use in many countries. Avian exposure to fipronil occurs mainly by ingesting contaminated insects or seeds. There is little information regarding the toxicological effects of fipronil in avian species and even less research documenting avian behavioural responses to fipronil ingestion. We examined the effects of a single oral dose of fipronil in northern bobwhite quail, the most fipronil-sensitive species tested to date, in respect to signs of intoxication and the metabolic fate of fipronil. Fipronil-treated birds did not eat or drink following pesticide administration, and as a result lost a significant amount of body mass. Treated birds also appeared withdrawn and did not respond to disturbance within the first hour after treatment. Identifiable signs of fipronil toxicity were not observed until at least 2 d after treatment. Chemical analyses indicated a difference between fipronil and fipronil–sulfone residue distribution and bioaccumulation, with significantly higher (30- to 1000-fold) tissue concentrations of the sulfone detected at all time points from 8 to 96 h post-dose in brain, liver and adipose tissues. Tissue sulfone concentrations increased significantly in fipronil-treated birds, peaking at 72 h post-dose. Body mass decreased at all time points in dosed birds. The coincidence of the particular intoxication symptoms with the time course of rise in brain sulfone levels after fipronil dosing gives insight into possible mechanisms of toxicity in this highly sensitive species.

Glutamatergic and GABAergic effects of fipronil on olfactory learning and memory in the honeybee

We investigated here the role of transmissions mediated by GABA and glutamate-gated chloride channels (GluCls) in olfactory learning and memory in honeybees, both of these channels being a target for fipronil. To do so, we combined olfactory conditioning with injections of either the GABA- and glutamate-interfering fipronil alone, or in combination with the blocker of glutamate transporter L-trans-Pyrrolidine-2,4-Dicarboxylicacid (L-trans-PDC), or the GABA analog Trans-4-Aminocrotonic Acid (TACA). Our results show that a low dose of fipronil (0.1 ng/bee) impaired olfactory memory, while a higher dose (0.5 ng/bee) had no effect. The detrimental effect induced by the low dose of fipronil was rescued by the coinjection of L-trans-PDC but was rather increased by the coinjection of TACA. Moreover, using whole-cell patch-clamp recordings, we observed that L-trans-PDC reduced glutamate-induced chloride currents in antennal lobe cells. We interpret these results as reflecting the involvement of both GluCl and GABA receptors in the impairment of olfactory memory induced by fipronil.

Horizontal Transfer of Insecticides in Laboratory Colonies of the Argentine Ant (Hymenoptera: Formicidae)

Five insecticides used by urban pest management professionals for ant control and three experimental insecticides were tested to determine whether these insecticides were horizontally transferred among individuals in colonies of Argentine ants, Linepithema humile (Mayr) (Hymenoptera: Formicidae). Ants were exposed to insecticide-treated sand for 1 min and then placed in a colony of untreated ants. Ants exposed to 20 and 40 ppm fipronil readily transferred the insecticide to other individuals in the colony, resulting in high mortality. Most of the transfer and subsequent mortality occurred within 4 d after exposure to treated ants. The other insecticides were not transferred, and ants exhibited mortality rates similar to that of the controls. Experiments in large foraging arenas demonstrated that necrophoresis was an important behavior facilitating the horizontal transfer of fipronil. When ants contacted contaminated corpses in the process of removing them to refuse piles, they received a lethal dose of fipronil and subsequently died. Fipronil-contaminated dead ants that were placed in the vicinity of the nest resulted in significantly higher mortality than did corpses placed in a distant foraging arena (30 cm away). Most of the dead ants accumulated in the vicinity of the nest rather than in the foraging arena, workers retrieving dead ants to refuse piles from the foraging arena. The position effect of insecticide-contaminated corpses relative to the nest and its implication for Argentine ant control are discussed.

Horizontal Transfer of Insecticides in Laboratory Colonies of the Argentine Ant (Hymenoptera: Formicidae)

Five insecticides used by urban pest management professionals for ant control and three experimental insecticides were tested to determine whether these insecticides were horizontally transferred among individuals in colonies of Argentine ants, Linepithema humile (Mayr) (Hymenoptera: Formicidae). Ants were exposed to insecticide-treated sand for 1 min and then placed in a colony of untreated ants. Ants exposed to 20 and 40 ppm fipronil readily transferred the insecticide to other individuals in the colony, resulting in high mortality. Most of the transfer and subsequent mortality occurred within 4 d after exposure to treated ants. The other insecticides were not transferred, and ants exhibited mortality rates similar to that of the controls. Experiments in large foraging arenas demonstrated that necrophoresis was an important behavior facilitating the horizontal transfer of fipronil. When ants contacted contaminated corpses in the process of removing them to refuse piles, they received a lethal dose of fipronil and subsequently died. Fipronil-contaminated dead ants that were placed in the vicinity of the nest resulted in significantly higher mortality than did corpses placed in a distant foraging arena (30 cm away). Most of the dead ants accumulated in the vicinity of the nest rather than in the foraging arena, workers retrieving dead ants to refuse piles from the foraging arena. The position effect of insecticide-contaminated corpses relative to the nest and its implication for Argentine ant control are discussed.

An in vitro feeding assay to test acaricides for control of hard ticks

Animal husbandry could not be practised over large areas of the planet without acaricides. The prevention of tick bite and the transmission of diseases requires the use of pesticides, but this contributes to the development of tick resistance against acaricides. This drives the quest for new molecules that target physiological processes crucial to tick survival. In vivo trials involve multiple repetitions because of inherent variations between host animals, requiring large amounts of test products and ticks. An in vitro alternative should permit the testing of the ability of a product to restrict attachment and feeding by ticks at precise doses. In this paper an in vitro feeding system is described where the European tick Ixodes ricinus L. feeds on blood through a cellulose rayon-reinforced silicone membrane. The membrane Shore hardness is modified to imitate the elastic retraction forces of skin that ensure the closing of tick penetration sites on the membrane to prevent bleeding. Tick attachment (75-100%) is achieved by adding chemical and mechanical stimuli to the membrane. Survival curves for different doses of fipronil and ivermectin tested with the method showed highly reproducible acaricide effects within 5-7 days. Significant effects are recorded down to ppb levels in blood. Standardised tests can be made with blood from the same donor animal or culture medium under the membrane.

Evaluation of black vine weevil (Otiorhynchus sulcatus) control strategies using Metarhizium anisopliae with sublethal doses of insecticides in disparate horticultural growing media

Abstract Metarhizium anisopliae V275 was efficacious against black vine weevil when used alone or in combination with sublethal doses of imidacloprid or fipronil. Both control strategies were effective in peat, coir, bark and peat blends with composted green waste. M. anisopliae can be applied as drench or premixed into growing media. The former gave marginally better control. Over 98% control was achieved when M. anisopliae was applied with 1/10 dilution (sublethal rates) of fipronil. Marginally lower control was achieved when M. anisopliae was used with 1/100 dilution (sublethal rate) of imidacloprid. However, the latter combination gave better control than imidacloprid used at the recommended rate. More conidia were recovered from larvae exposed to sublethal doses of fipronil than imidacloprid and may explain the higher pest control since mortality is dose-related.