Efficacy of 1% Permethrin and 2% Permethrin Combined with 1% Piperonyl Butoxide against Rhipicephalus sanguineus Ticks in Naturally Infested Dogs

The aim of the present study was to compare the ixodicide efficacy of 1% permethrin applied as a shampoo-bath in single treatment and a combined formulation of 2% permethrin + 1% piperonyl butoxide applied as powder in a single treatment against Rhipicephalus sanguineus ticks, in naturally infested dogs. Eighteen adult cross-mixed dogs, harboring to at least 20 ticks/animal, were selected on pre-treatment live tick counts. On day 0, the dogs were then divided in 3 groups of 6 animals each, for treatment: Group 1 (G1) received 1% permethrin applied as shampoo-bath (approximately 10 ml/kg bw), according to label instructions. G2 was treated with 2% permethrin + 1% piperonyl butoxide applied as powder, and rubbed as massage on the body, as recommended by the producer. G3 served as a non-treated control. After the treatment, the dogs were checked out for live ticks counting on days 0, 1, 7, and 14. The ixodicide efficacy was assessed based on the percentage reductions of ticks on the days analyzed post-treatment, with respect to the untreated control. Results showed that permethrin alone removed ticks on 81%, 93.1% and 89.6% for days 1, 7 and 14, respectively. The combined formulation of 2% permethrin + 1% piperonyl butoxide exerted a better efficacy of 98.8, 97.7 and 97.0%, respectively. It is concluded that this combined formulation was highly effective at rapidly repelling and killing Riphicephalus sanguineus ticks on naturally infected dogs, showing a significant effect on days 1, 7 and 14 after tick exposure. The sustained high and quick level of efficacy of this combination may well interfere with the transmission of Riphicephalus sanguineus ticks. It is concluded that the sustained high level of efficacy of the permethrin + piperonyl butoxide combination may well exert an additive effect and could interfere with the transmission of Riphicephalus sanguineus ticks.

The Entomological Efficacy of Piperonyl Butoxide (PBO) Combined with a Pyrethroid in Insecticide-Treated Nets for Malaria Prevention: A Village-Based Cohort Study Prior to Large-Scale Deployment of New Generation Mosquito Nets in Burkina Faso

The efficacy of insecticide-treated nets (ITNs) is increasingly compromised by the prevalence of malaria vectors resistant to pyrethroids. In response to this issue, a new generation of ITNs has been developed that incorporate synergistic components, such as piperonyl butoxide (PBO). The purpose of this study is to provide entomological evidence for the efficacy of a PBO-based ITN brand at the village level, serving as a basis for decision-making before large-scale net deployment. During the high malaria transmission period, ITNs were distributed in each group and vector sampling was conducted biweekly in selected households. Bionomic data were collected to assess the resistance of wild An. gambiae populations to various chemical insecticides. There was a significant disparity in total An. gambiae sl. collected between the ITN arms, the intervention arm (ITN arms), and the control arm (P = 0.003). An. coluzzi was identified as the predominant species in the study area, as confirmed by PCR analysis. Analysis of the blood-feeding inhibition rate revealed that 100% permethrin + PBO ITN exhibited significantly greater inhibition than 66.81% permethrin only ITN. According to the log-time probit regression analysis, permethrin exhibited a knockdown time of 256 min without synergists, while it decreased to 139 min (P = 0.001) when pre-exposed to PBO. The evidence from this trial supports the use of PBO ITNs over standard ITNs for pyrethroids to combat pyrethroid resistance and improve protection against malaria for both individuals and communities, particularly in areas with high pyrethroid resistance.

Susceptibility to temephos,permethrin and deltamethrin of Aedes aegypti(Diptera:Culicidae) from Muang district,Phitsanulok Province,Thailand

Objective:To investigate the susceptibility to temephos.permethrin and deltamethrin of Aedes aegypti Ae.aegypti),collected from areas with high incidence of dengue hemorrhagic fever cases in Phitsanulok Province.Thailand.Methods:The F1 progenies of Ae.aegypti colony,originated from five sub-districts including Aranyik.Hua Ro,Nai Muang.Ban Krang and Tha Pho,were used in the bioassays following the procedures of World Heath Organization.For larval bioassay.the late third or early fourth-instar lanae were tested with different concentrations of temephos.For adult bioassay.the females were exposed to 0.75%permethrin or 0.05%deltamethrin.LC_(50)value and mortality rate were analyzed to compare the insecticide susceptibility of the larvae and the adults in each area,respectively.Results:The LC_(50) value of temephos for the larvae from Aranyik.Hua Ro.Nai Muang,Ban Krang and Tha Pho sub-districts was 0.017.0.017.0.026.0.061.and 0.113 ppm,respectively.For permethrin,the highest morlalitv rate(86.84%) was found in the mosquitoes from Aranyik but the others were more resistant with the lower morlality rates(16.00-42.67%).The adult morlality rates after exposing to dellaiiielhrin were higher(82.34-98.67%) in all areas.Conclusions:Ae.acgypti larvae were still susceptible to temephos.Conversely,most tested adults tended to resist the penmethrin and deltamethrin.

Comparative susceptibility to permethrin of two Anopheles gambiae s.l.populations from Southern Benin,regarding mosquito sex,physiological status,and mosquito age

Objective:To investigate what kind of mosquito sample is necessary for the determination of insecticide susceptibility in malaria vectors.Methods:Larvae and pupae of Anopheles gambiae s.l.(An.gambiae)mosquitoes were collected from the breeding sites in Littoral and Oueme departments.The Centers for Disease Control and Prevention(CDC)susceptibility tests were conducted on unfed male and female mosquitoes aged 2-5 days old.CDC susceptibility tests were also conducted on unfed,blood fed and gravid female mosquitoes aged2-5 days old.These susceptibility tests were also conducted on unfed and blood fed female mosquitoes aged 2-5 days old and 20 days old.CDC biochemical assay using synergist was also carried out to detect any increase in the activity of enzyme typically involved in insecticide metabolism.Results:Female An,gambiae Ladji and Sekandji populations were more susceptible than the males when they were unfed and aged 2-5 days old.The mortality rates of blood fed female An.gambiae Ladji and Sekandji populations aged 2-5 days old were lower than those obtained when females were unfed.In addition,the mortality rales of gravid female An.gambiae Ladji and Sekandji populations aged2-5 days old were lower than those obtained when they were unfed.The mortality rate obtained when female An.gambiae Sekandji populations were unfed and aged 20 days old was higher than the one obtained when these populations were unfed and aged 2-5 days old.The results obtained after effects of synergist penicillin in beeswax on F1 progeny of An.gambiae Ladji populations resistant to permethrin showed that mono-oxygenases were involved in permethrin resistant F1 progeny from Ladji.Conclusions:The resistance is a hereditary and dynamic phenomenon which can be due to metabolic mechanisms like overproduction of detoxifying enzymes activity.Many factors influence vector susceptibility to insecticide.Among these factors,there are mosquito sex,mosquito age,its physiological status.Therefore,it is useful to respect the World Health Organization criteria in the assessment of insecticide susceptibility tests in malaria vectors.Otherwise,susceptibility testing is conducted using unfed female mosquitoes aged 3-5 days old.Tests should also be carried out at(25±2)°C and(80±10)%relative humidity.

The genetic architecture of insecticide resistance within a natural population of <i>Drosophila melanogaster</i>

The dynamics of genetic variation in susceptibility to insecticides within a natural population of Drosophila melanogaster (Meigen) at Katsunuma (Yamanashi Prefecture, Japan) was examined. Two resistance factors for three organophosphate insecticides (OPs), a resistant-type acetylcholinesterase (AChE) and a cytochrome P450 monooxygenase (cytochrome P450), have already been suggested to be involved within the Katsunuma population. In this study, genetic variances were estimated for susceptibility to other classes of chemicals than OPs, permethrin (a pyrethroid) and dichloro-diphenyl-trichloroethane (DDT;an organo-chlorine), which existed simultaneously with genetic variances for susceptibility to OPs. Analyses of variance for susceptibility to permethrin and DDT showed highly significant variation among isofemale lines from the Katsunuma population, and the genetic variances for susceptibility to each insecticide fluctuated differently during this period. The impacts of fluctuations of genetic variation in susceptibility to one class of insecticides on genetic variation in susceptibility to other classes of insecticides existing simultaneously within the natural population were discussed.

Phomalactone as the Active Constituent against Mosquitoes from <i>Nigrospora spherica</i>

The culture filtrate of a plant pathogenic fungus that infected Zinnia elegans and Hydrangea macrophylla was investigated for mosquitocidal constituents by bioassay guided isolation. The fungus responsible for the pathogenic effects on Zinnia elegans and Hydrangea macrophylla plants had been identified as Nigrospora spherica by molecular techniques. The mosquito adulticidal constituent in the culture filtrate was identified as phomalactone by spectroscopic techniques. Laboratory bioassays showed that phomalactone had larvicidal activity against permethrin susceptible and resistant Aedes aegypti larvae and topical adulticide activities on permethrin susceptible and resistant Aedes aegypti and Anopheles quadrimaculatus mosquitoes. Phomalactone was effective as a topical adulticide against the standard Orlando reference strain of Ae. aegypti with an LD50 of 0.64 μg/org. Activity against An. quadrimaculatus was 0.20 μg/org.

Changes in Myelinated Nerve Fibers and Skeletal Muscle of Rats Exposed to High Doses of Permethrin

Neurological signs and segmcntal demyelination in a cervical nerve were observed in rats treated orally with permethrin (300 mg/kg/day) for 5 days. Inflammatory and degenerative signals were recorded in the diaphragm muscle. These effects were more intense with the trade grade than with the technical grade product. The possible influence of the percentage of cisitrans isomers on the intensity of the observed effects is discussed. 5 imi Academic Press.Inc.

Wash resistance and bio-efficacy of Olyset~? Plus, a long-lasting insecticide-treated mosquito net with synergist against malaria vector, Anopheles stephensi

Objective: To determine the wash resistance of Olyset~? Plus using World Health Organization Pesticide Evaluation Scheme standard washing procedure and to assess the value of knock down and mortality rates of Anopheles stephensi at different regimens of long lasting insecticide treated nets washings.Methods: The study was conducted at the Bioassay Laboratory of Culicidae Insectary,School of Public Health, Tehran University of Medical Sciences, Iran. The net was made of polyester impregnated with permethrin and piperonyl butoxide at a ratio of 2:1. The washing resistance was assessed using Le Chat?soap and a shaker incubator set at a speed of 155 r/min, 30℃ for 10 min. The cone bioassay test was carried out according to World Health Organization recommended guideline with tolerant field strain of female Anopheles stephensi to pyrethroids.Results: The knockdown and mortality rates of female mosquitoes exposed to Olyset~? Plus from un-washed nets to 2 washings were 79.7% and 88.8% respectively. Mortality was dropped to zero while active ingredient estimated 0.532 mg/100 cm^2 to 0.481 mg/100 cm^2 after 15 washings. A positive correlation was seen between residues of permethrin on nets, knockdown rate and mortality rate of female Anopheles stephensi exposed to different regimes of washed Olyset~? Plus(r = 0.954, P = 0.001).Conclusions: It is recommended that a preliminary survey conducted on resistance level of Anopheles vectors before the distribution of Olyset~? Plus in malaria endemic communities.

Effect of Particle Size and Pesticide Contamination on Preference and Ingestion Rates by the Tropical Freshwater Shrimp Xiphocaris elongata

In tropical streams, freshwater shrimp are essential to preserve the structure and function of lotic ecosystems. Shredder shrimp play a fundamental role in organic matter decomposition because these feed on detritus. In addition, they are especially important organisms as they connect all trophic levels at food webs. In natural environments, decaying leaf material may accumulate contaminants, including insecticides and herbicides. At these, shredder shrimp can be exposed to these contaminants through ingestion of leaf litter material. The objectives of this study were to evaluate if the shredder shrimp Xiphocaris elongata display a preference for feeding on different plant species and leaf size areas while also assessing their consumption of leaves contaminated with pesticides. We also evaluated acetylcholinesterase (AChE) activity as a possible biomarker of pesticide contamination using an immunofluorescence and microscopy imaging approach. Our results revealed that the leaf area and plant species more appropriate for future toxicological studies is Spathodea campanulata leaves, with a leaf area of 0.65 cm2. This study also showed that sublethal concentrations of malathion and permethrin in leaves seem to have a significant effect on the ingestion rates of X. elongata, which suggests that the presence of these contaminants influenced feeding behavior. Immunofluorescence in cephalothorax ganglia showed a decline in AChE activity when the sublethal dose of malathion and permethrin increased. The observed results suggest that AChE activity can be used as a biomarker to detect and assess permethrin and malathion exposure on shredder shrimp.

Comparative Efficacy of Three Commercial Ectoparasiticides against Fleas in Naturally Infested Dogs

Fleas that infest pets are considered important parasites of both animals and humans. These insects cause irritation and can also transmit zoonotic diseases. Research has led to a rapid expansion in the development of flea control products. In the face of a market that offers dozens of commercial ectoparasiticides for dogs and cats, pet owners and veterinarians must be provided with evidence to support their decision to select a product to control fleas. To compare the efficacy of three commercially available products against companion animal fleas, a trial was conducted on naturally-infested dogs in order to validate their pulicidal activity. Thirty-two flea-infested dogs with fleas were divided into 4 groups (n = 8) for each treatment. Group 1 received one pipette of permethrin as a spot-on dose of 650 mg/ml. The second group received 9.7% fipronil as a spot-on formulation. Group 3 was treated with a spot-on formulation of permethrin 7.40% plus piperonyl butoxide at 7.40%. Group 4 remained as the untreated control. Fleas of all experimental dogs were examined and counted on days 0, 3, 7, 14, 21 and 28 to determine the percentage of flea reduction. Results showed a 100% efficacy for all tested products. Identified species were: Ctenocephalides felis (75.7%), Ctenocephalides canis (15.9%) and Pulex irritans (9.5%). Based on these results, it was concluded that the three anti-flea products evaluated under the conditions of this study, produced an excellent efficacy as from the third day after treatment.

Evaluation of the Efficacy and Safety of 1%Sodium Chloride(LiceFreee Spray)against 1%Permethrin Creme Rinse on Head Lice Infested Individuals

Head lice are a public health issue, and resistance to available over-the-counter pediculicides is a concern. The objective of this randomized study was to evaluate the pediculicidal activity and safety of 1% Sodium Chloride spray (0.1709 M), (LiceFreee Spray&#174) compared to the current recommended treatment for head lice with 1% Permethrin Crème Rinse. Forty-two subjects were randomized equally into Sodium Chloride or Permethrin group. Products were applied to hair according to the label instructions. After application of the products at Day 1 and Day 8, pediculicidal efficacy and safety were assessed at Day 1, Day 8 and Day 15. Second treatment was only applied on Day 8 to individuals with observed live lice using the same products and protocols as Day 1. Proportion of lice free subjects per group, the reduction in number of live lice per head and adverse effects were recorded after each visit. The results showed significantly higher pediculicidal activity for Sodium Chloride spray (85%) as compared to Permethrin (45%) at Day 15 (p 0.05). Similar numbers of lice per head (21.76 range 10 to 68 versus 21.29 range 10 to 60 for Sodium Chloride and Permethrin groups, respectively) were observed for individuals at Day 1. At Day 15, lice per head infested reduced to 0.55 ± 1.50 in Sodium Chloride spray group compared to 5.45 ± 7.91 in the Permethrin group (p 0.01). No serious adverse effects were observed in both groups. Sodium Chloride spray had superior efficacy to 1% Permethrin Crème Rinse in treating head lice and is a safe and excellent alternative to the current recommended treatment.

Negative cross-resistance of a pyrethroid-resistant Drosophila mutant to Phryma leptostachya-derived haedoxan A

Voltage-gated sodium channels are the primary target of pyrethroid insecticides.Mutations in sodium channel confer knockdown resistance(kdr)to pyrethroids in various arthropod pests.Haedoxan A(HA)is the major insecticidal component from Phryma leptostachya.It has been shown that HA alters electrical responses at the Drosophila neuromuscular junction and modifies the gating properties of cockroach sodium channels expressed in Xenopus oocytes.However,whether sodium channel mutations that confer pyrethroid resistance also affect the action of HA is unknown.In this study,we conducted bioassays using HA and permethrin in two Drosophila melanogaster strains:w^(1118),an insecticide-susceptible strain,and para^(tsl),a pyrethroid-resistant strain due to a I265N mutation in the sodium channel,and identified a new case of negative cross-resistance(NCR)between permethrin and HA.Both para^(tsl) larvae and adults were more resistant to permethrin,as expected.However,both para^(tsl) larvae and adults were more sensitive to HA compared to w^(1118).We confirmed that the I265N mutation reduced the sensitivity to permethrin of a Drosophila sodium channel variant,DmNa_(v)22,expressed in Xenopus oocytes.Interestingly,the I265N mutation also abolished the effect of HA on sodium channels.Further characterization showed that I265 on the sodium channels is critical for the action of both pyrethroids and HA on sodium channels,pointing to an overlapping mode of action between pyrethroids and HA on the sodium channel.Overall,our results suggest an I265N-independnt mechanism(s)in para^(tsl) flies that is responsible for the NCR between permethrin and HA at the whole insect level.

Molecular and functional characterization of three novel carboxylesterases in the detoxification of permethrin in the mosquito, Culex quinquefasciatus

Carboxylesterases (CarEs) belong to a super family of multifunctional enzymes associated with the degradation of endogenous and exogenous compounds. Many insect CarEs are known to play important roles in catalyzing the hydrolysis of organophosphates (OPs), carbamates, and synthetic pyrethroids (SPs). The elevation of esterase activity through gene amplification and overexpression of estα2 and estβ2 genes contributes to the development of resistance to OP insecticides in the mosquito Culex quinquefasciatus. Three additional CarE genes are upregulated in permethrin-resistant Cx. quinquefasciatus according to an RNA-seq analysis, but their function remains unknown. In this study, we, for the first time, characterized the function of these three novel genes using in vitro protein expression, an insecticide metabolism study and molecular docking analysis. All three CarE genes were significantly overexpressed in resistant mosquito larvae, but not adults, compared to susceptible strain. No gene copy differences in these three genes were found in the mosquitoes tested. In vitro high-performance liquid chromatography (HPLC) revealed that CPIJ018231, CPIJ018232, and CPIJ018233 metabolized 30.4% ± 2.9%, 34.7% ± 6.8%, and 23.2% ± 2.2% of the permethrin, respectively. No mutations in resistant strains might significantly affect their CarE hydrolysis ability. A docking analysis further confirmed that these three CarEs from resistant strain all potentially metabolize permethrin. Taken together, these three carboxylesterase genes could play important roles in the development of permethrin resistance in Cx. quinquefasciatus larvae through transcriptional overexpression, metabolism, and detoxification.

Dynamics and mechanisms of permethrin resistance in a field population of the horn fly, Haematobia irritans irritans

A study was conducted at the Pressler ranch, near Kerrville, Texas, USA between 2002 and 2006 to determine the dynamics and mechanisms of resistance to permethrin in a field population of the horn fly, Haematobia irritans irritans （L.）. Changes of resistance to pyrethroid insecticide associated with use of a pour-on formulation of cyfluthrin in 2002 and use of diazinon ear tags in subsequent years were studied using a filter paper bioassay technique and a polymerase chain reaction assay that detects two sodium channel mutations, kdr and super-kdr resistance alleles. A maximum of 294-fold resistance to permethrin was observed in the summer of 2002. A significant decrease in the resistance level was observed in spring 2003, and resistance continued to decline after animals were treated with diazinon ear tags. In response to pyrethroid treatments, the allelic kdr and super-kdr frequency increased from 56.3% to 93.8% and from 7.5% to 43.8%, respectively in 2002, and decreased significantly in 2003 when the pyrethroid insecticide was no longer used to treat animals. Females were found to have a higher allelic super-kdr frequency than males in 2002, while no difference was detected between males and females in the allelic kdr frequency. There was a significant positive correlation between frequencies of the sodium channel mutations and levels ofpermethrin resistance, suggesting that the sodium channel mutations, kdr and super-kdr, are the major mechanisms of resistance to pyrethroids in this horn fly population. Results of synergist bioassays also indicated possible contributions of two metabolic detoxification mechanisms, the mixed function oxidases （MFO） and glutathione S-trans- ferases （GST）. Compared to a horn fly infestation of an untreated herd, treatments with the pyrethroid pour-on formulation failed to control horn flies at the Pressler ranch in 2002. Sustained control of horn flies was achieved with the use of diazinon ear tags in 2003 and subsequent years.

Bioefcacy and durability of Olyset® Plus, a permethrin and piperonyl butoxide-treated insecticidal net in a 3-year long trial in Kenya

Background:Long-lasting insecticide nets(LLINs)are a core malaria intervention.LLINs should retain efficacy against mosquito vectors for a minimum of three years.Efficacy and durability of Olyset®Plus,a permethrin and piperonyl butoxide(PBO)treated LLIN,was evaluated versus permethrin treated Olyset®Net.In the absence of WHO guidelines of how to evaluate PBO nets,and considering the manufacturer’s product claim,Olyset®Plus was evaluated as a pyrethroid LLIN.Methods:This was a household randomized controlled trial in a malaria endemic rice cultivation zone of Kirinyaga County,Kenya between 2014 and 2017.Cone bioassays and tunnel tests were done against Anopheles gambiae Kisumu.The chemical content,fabric integrity and LLIN survivorship were monitored.Comparisons between nets were tested for signifcance using the Chi-square test.Exact binomial distribution with 95%confdence intervals(95%CI)was used for percentages.The WHO efcacy criteria used were≥95%knockdown and/or≥80%mortality rate in cone bioassays and≥80%mortality and/or≥90%blood-feeding inhibition in tunnel tests.Results:At 36 months,Olyset®Plus lost 52%permethrin and 87%PBO content;Olyset®Net lost 24%permethrin.Over 80%of Olyset®Plus and Olyset®Net passed the WHO efcacy criteria for LLINs up to 18 and 12 months,respec‑tively.At month 36,91.2%Olyset®Plus and 86.4%Olyset®Net survived,while 72%and 63%developed at least one hole.The proportionate Hole Index(pHI)values representing nets in good,serviceable and torn condition were 49.6%,27.1%and 23.2%,respectively for Olyset®Plus,and 44.9%,32.8%and 22.2%,respectively for Olyset®Net but were not signifcantly diferent.Conclusions:Olyset®Plus retained efcacy above or close to the WHO efcacy criteria for about 2 years than Olyset®Net(1–1.5 years).Both nets did not meet the 3-year WHO efcacy criteria,and showed little attrition,comparable physical durability and survivorship,with 50%of Olyset®Plus having good and serviceable condition after 3 years.Better community education on appropriate use and upkeep of LLINs is essential to ensure efectiveness of LLIN based malaria interventions.

Permethrin resistance variation and susceptible reference line isolation in a field population of the mosquito, Culex quinquefasciatus （Diptera： Culicidae）

This study examines the genetic variations and mechanisms involved in the development of permethrin resistance in individual mosquitoes from a field population of Culex quinquefasciatus, HAmCq^G0, and characterizes susceptible reference lines of mosquitoes with a similar genetic background to the field HAmCq^G0 strain. Six upregulated cytochrome P450 genes, CYP9M10, CYP9J34, CYP6P14, CYP9J40, CYP6AA7, and CYP4C52v1, previously identified as being upregulated in the larvae of resistant HAmCq68 mosquitoes were examined in the larvae of 3 strains （susceptible S-Lab, parental HAmCq^G0 and permethrin-selected highly resistant HAmCq68） and 8 HAmCq^G0 single- egg raft colonies, covering a range of levels of susceptibility/resistance to perrnethrin and exhibiting different variations in the expression of A and/or T alleles at the L-to-F kdr locus of the sodium channel. The 2 lines with the lowest tolerance to permethrin and bearing solely the susceptible A allele at the L-to-F kdr locus of the sodium channels, from colonies Cx_SERC5 and Cx_SERC8, showed lower or similar levels of all 6 of the P450 genes tested compared with the S-Lab strain, suggesting that these 2 lines could be used as the reference mosquitoes in future studies characterizing insecticide resistance in HAmCq mosquitoes. This study also provides a detailed investigation of the mechanisms involved in insecticide resistance in individuals within a population： individuals with elevated levels of resistance to permethrin all displayed one or more potential resistance mechanisms-either elevated levels of P450 gene expression, or L-to-F mutations in the sodium channel, or both.

Toxicological and histopathological effects of Dennettia tripetala seed used as grain protectant,food,and medicine

The study revealed the safety of Dennettia tripetala seed on man and the environment.Adult male rats weighing 0.158-0.168 kg housed in standard cages with free access to food and water were used for the experiments.The median lethal dose(LD50)was estimated using revised up and down procedure.The LD50 for D.tripetala seed extract was 5785 mg/kg and this evoked paralysis in rats for 4 days coupled with discharge from the eyes and eventual death.The least weight gain by the animals administered 75%seed powder of D.tripetala(4338.75 mg/kg)was an index of high powder concentration,whereas the weight loss experienced by group V animals is strongly attributed to chemical assault by permethrin designated as a standard insecticide.The high values of alanine aminotransferase(ALT),aspartate aminotransferase(AST),and urea in the groups administered 75%permethrin powder(4338.75 mg/kg)and 75%D.tripetala seed powder is an index of liver and kidney injury and dysfunction.The presence of normal serum levels of ALT,AST,alkaline phosphatase,and creatinine;normal liver and kidney structures;and normal weight gains in the animal groups fed basal diet(control)and basal diet plus 25%D.tripetala is a strong indication that 25%D.tripetala seed powder(1446.25 mg/kg)supplementation is not toxic to the liver and kidney and therefore supports normal organ functions.The LD50 recorded strongly indicates that D.tripetala has a moderately high safety margin.Supplementation of less than 50%(2892.5 mg/kg)is recommended in the safe use of the plant material as grain protectant,food,and medicine.The botanical insecticide,D.tripetala,is safer than the conventional synthetic insecticide,permethrin,on account of the latter showing evidence of kidney damage.