Efficacy of a pour-on formulation of doramectin against lice, mites, and grubs of cattle

OBJECTIVE: To determine effectiveness of a pour-on formulation of doramectin against Damalinia bovis, Haematopinus eurysternus, Linognathus vituli, Solenopotes capillatus, Chorioptes bovis, Sarcoptes scabiei, Hypoderma bovis, and Hypoderma lineatum. ANIMALS: Cattle of various ages with naturally acquired or artificial infestations with 1 or more species of lice, mites, or grubs. PROCEDURE: In 10 louse and 6 mite studies, cattle were treated with doramectin (500 microg/kg, topically) on day 0, and parasite counts were performed approximately weekly from days 0 to 35. In 6 grub studies, cattle expected to harbor Hypoderma spp were treated before emergence of warbles. After warbles began to emerge, they were counted every 2 weeks, and grubs were collected and identified by species. RESULTS: Burdens of D bovis, H eurystemus, L vituli, and S capillatus on doramectin-treated cattle were 0 by 28 days after treatment. Burdens of C bovis and S scabiei decreased to 0 in naturally infested cattle and approximately 0 in artificially infested cattle by day 14 to 15. In grub studies, 107 of 136 control cattle had warbles, whereas 2 of 136 doramectin-treated cattle had 1 warble each, which represented a cure rate of 98.5%. CONCLUSION AND CLINICAL RELEVANCE: One topical application of doramectin was highly efficacious against common species of lice, mites, and grubs known to affect performance, health, and appearance of cattle.     

Persistent efficacy of doramectin topical solution against induced infestations of Bovicola bovis and Solenopotes capillatus.

A study was conducted in two locations, Wyoming and Wisconsin, USA, to evaluate the persistent efficacy of doramectin topical solution at a dose rate of 500 microg/kg body weight against artificially induced infestations of Bovicola bovis and Solenopotes capillatus on cattle. At each location, lice-free beef calves were individually housed and randomly allocated to treatment. Both B. bovis and S. capillatus were transferred from untreated donor animals to doramectin-treated cattle at the end of 35, 63, 91 or 126 day post-treatment periods. Cattle treated with a saline pour-on served as the control. Based on the geometric means of lice counts 2 weeks following transfer, the persistent efficacy of a single treatment with doramectin topical solution against induced infestations of B. bovis was 100.0, 100.0, 99.5, and 100.0% at post-treatment days of 35, 63, 91, and 126, respectively. Persistent efficacy against induced infestations of S. capillatus, for the same intervals, were 100.0, 94.9, 86.3, and 74.9%.     

Prevalence of sucking and chewing lice on cattle entering feedlots in southern Alberta

Beef calves from 2 sources entering southern Alberta feedlots in the winters of 1997-98 and 1998-99, were surveyed for the presence of lice. A random sample of multiple source (MS), that is, auction market-derived, calves entering commercial feedlots and single source (SS) calves entering a backgrounding feedlot were examined for the presence of lice at entry to the feedlot. A standardized examination, which involved hair-part examination of 8 louse predilection sites, was conducted on each selected calf to determine prevalence and intensity of infestation. The long-nosed sucking louse, Linognathus vituli, was the most commonly encountered species. This species infested from 57.8% to 95.6% of the calves selected from both MS and SS calves during both winters. Louse index values, indicating intensity of infestation, for L. vituli ranged from 1 to 243 lice per animal. The chewing louse, Bovicola bovis, was present on MS and SS calves only in the winter of 1998-99. The louse index values for B. bovis ranged from 1 to 230 lice per animal. Mixed infestations of the L. vituli and B. bovis were common. The little blue cattle louse, Solenopotes capillatus, was present only on the SS calves in the winter of 1997-98. The short-nosed sucking louse, Haematopinus eurysternus, was present at very low intensities, 1-2 lice per animal, on 2.6% to 4.4% of the MS calves during both winters. Comparison of results from the current study with published literature suggests that efforts to determine the economic impact of louse infestations are confounded by the lack of a uniform method to assess louse population levels.     

Efficacy of a 1% lambdacyhalothrin cattle Pour-on (Saber) against sucking and biting lice infesting beef cattle

Four studies were conducted to determine the efficacy of a 1% lambdacyhalothrin pour-on (Saber Pour-on, Schering-Plough Animal Health) for control of sucking lice (Linognathus vituli, Haematopinus eurysternus, Solenopotes capillatus) and biting lice (Damalinia bovis) on beef cattle. Seventy-four mixed-breed cattle naturally infested with one or more species of lice at locations in Wisconsin, Nebraska, Oklahoma, and North Dakota were included in the study. Pretreatment lice samples were taken and identified by genus with the exception that sucking lice were not identified by genus at the North Dakota site. In January or February, half of the cattle at each location received a single application of 1% lambdacyhalothrin pour-on at 10 ml per head for cattle weighing less than 273 kg (600 lb) or 15 ml per head for cattle weighing 273 kg or more. The other cattle at each site served as untreated controls. Lice on designated body areas were counted 2, 4, 6, and 8 weeks after treatment, and the sum of all lice observed on each counting area was reported for each animal. The collective efficacy 6 weeks after treatment against three species of sucking lice (L. vituli, H. eurysternus, S. capillatus) was 88.4% at the Wisconsin site, 92.0% at the North Dakota site, and 100% at the Nebraska and Oklahoma sites. The 1% lambdacyhalothrin pour-on eliminated all biting lice within 2 weeks after treatment, and no biting lice were detected 8 weeks after treatment. A single treatment of 1% lambdacyhalothrin pour-on administered when lice populations were highest (January or February) provided effective season-long control of both biting and sucking lice on cattle.     

Persistent activity of moxidectin pour-on and injectable against sucking and biting louse infestations of cattle

To evaluate the persistent activity of pour-on and injectable moxidectin against natural challenge by sucking (predominantly Linognathus vituli) and chewing (Bovicola bovis) cattle lice, 96 mixed-breed calves that had been treated to remove all lice were blocked by body weight and randomly allocated to three treatments: untreated control, moxidectin at 500 microg/kg by topical application and moxidectin at 200 microg/kg by subcutaneous injection. Twelve pens were blocked into groups of four and randomly allocated to four challenge times: 14, 21, 28 and 35 days post-treatment. Treatment groups were assigned to challenge pens randomly. Two donor calves, with demonstrated infestations of both sucking and chewing lice, were introduced into each pen containing eight principal calves at the start of each challenge time. Donors remained in the challenge pen for 7 days. Principal calves were examined for lice, 7, 14, 21 and 28 days after donor removal using a standardized hair-parting technique. Moxidectin injectable prevented re-infestation with L. vituli for up to 42 days, but did not provide persistent activity against B. bovis longer than 35 days post-treatment. Moxidectin pour-on demonstrated persistent activity against both B. bovis and L. vituli for 42 days.     

Persistent activity of moxidectin long-acting injectable formulations against natural and experimentally enhanced populations of lice infesting cattle

A study was conducted under a common protocol in Wisconsin and Wyoming, USA, to evaluate therapeutic and persistent efficacy of two long-acting injectable formulations of moxidectin against lice populations infesting cattle. At each site, 30 beef calves were blocked into groups of three based on naturally acquired Linognathus vituli populations, then randomly assigned to treatments within blocks. Treatments, injected subcutaneously into the proximal third of the ear on Day 0, included saline, a long-acting oil-based formulation containing 10% moxidectin given at the rate of 1 mg moxidectin/kg body weight (M10/1.0), or a long-acting oil-based formulation containing 15% moxidectin given at the rate of 0.75 mg moxidectin/kg b.w. (M15/0.75). Species of sucking and chewing lice were quantified on nine predilection sites before treatment, then 28, 63, 98, 133 and 168 days after treatment. During intervals between lice counts after Day 28, study animals from the three treatment groups were commingled for 32 days with two lice-free sentinels plus four to six seeder calves with infestations of both sucking and chewing lice. Following each 32-day commingling interval, seeder and sentinel animals were removed, and principal animals were sorted into pens by treatment. Lice were quantified on sentinel animals on the day of removal, and lice were quantified on principal study animals 3 days after removal of sentinel and seeders. Moxidectin was generally not efficacious against Bovicola bovis in the injectable formulations tested, whereas Haematopinus eurysternus infestations were inadequate to judge product effectiveness. Based on geometric means, both M15/0.75 and M10/1.0 provided statistically significant therapeutic efficacy against existing infestations of L. vituli and Solenopotes capillatus (100% efficacy on Day 28), and provided persistent protection against reinfestation with L. vituli and S. capillatus (efficacy >97%) for at least 133 days following treatment.     

Therapeutic and persistent efficacy of spinosad applied as a pour-on or a topical spray against natural infestations of chewing and sucking lice on cattle

Studies were conducted in Wisconsin and Illinois, USA, to assess and compare the therapeutic and persistent efficacy of spinosad when applied as either a pour-on or topical spray and compared with cyfluthrin pour-on and coumaphos topical spray for controlling natural infestations of chewing (Bovicola bovis) and sucking (Linognathus vituli, Solenopotes capillatus and Haematopinus eurysternus) lice on cattle. Thirty-five animals at each trial site were blocked according to pre-treatment lice counts and randomly allocated to one of five treatment groups: single treatments of spinosad (25 g/L), diluted with water to 0.04% active ingredient and applied as a whole-body topical spray; spinosad (25 g/L) applied as a neat pour-on at 2 mg/kg body weight; Co-Ral Emulsifiable Livestock Insecticide (5.8% coumaphos), diluted to 0.03% active ingredient and applied as a whole-body topical spray; CyLence Pour-on Insecticide (1% cyfluthrin), applied as a neat pour-on at the manufacturer's recommended use rate for lice; and untreated control. Both spinosad treatments and cyfluthrin provided > or =96% control of B. bovis for up to 7 weeks, whereas the efficacy of coumaphos dropped to <90% after week 5 at one site. Spinosad spray had the best therapeutic and residual control of all treatments against L. vituli, > or =98% for at least 5 weeks at both sites, compared with 3 weeks for coumaphos at one site. Spinosad and coumaphos sprays provided 100% control of S. capillatus for at least 8 weeks compared with > or =97% control for spinosad and cyfluthrin pour-on treatments over the same interval. While H. eurysternus burden was low and limited to one study site, all four treatments provided 100% control for at least 6 weeks. These studies showed that topically applied spinosad provided a high degree of therapeutic and residual control against both sucking and chewing lice.     

Therapeutic efficacy of zeta-cypermethrin pour-on for the treatment of biting and sucking lice in cattle under field conditions

OBJECTIVE: To assess the efficacy of zeta-cypermethrin pour-on to control cattle lice. DESIGN: Five field trials in south-eastern Australia. PROCEDURE: Zeta-cypermethrin pour-on, deltamethrin pour-on and pour-on vehicle were applied to groups of 10 cattle. Lice were counted before treatment and 14, 28, 42 and 56 days after treatment. RESULTS: Zeta-cypermethrin pour-on given at 2.5 mg/kg was equivalent to, or marginally more effective than a deltamethrin pour-on at 0.75 mg/kg. It eliminated B bovis and H eurysternus and gave good control of L vituli and S capillatus. Zeta-cypermethrin at 1 mg/kg gave good control of B bovis and H eurysternus but was not satisfactory against L vituli and S capillatus. CONCLUSION: Zeta-cypermethrin pour-on, given at 2.5 mg/kg, is an effective treatment for cattle lice control. Zeta-cypermethrin, and other synthetic pyrethroid pour-ons, are the treatment of choice to control B bovis.     

Anaemia and mortality in calves infested with the long-nosed sucking louse (Linognathus vituli)

Young calves which died on three dairy farms in England and Wales had generalised pallor and heavy infestations of the long-nosed sucking louse, Linognothus vituli. Surviving calves had packed-cell volumes, haemoglobin concentrations and red blood cell counts below the reference ranges, consistent with anaemia, and they were also heavily infested with lice. No other causes of anaemia were identified. It is proposed that heavy infestations with L. vituli should be considered when investigating the cause of anaemia in calves.     

Impact of the duration of control of cattle lice with eprinomectin on leather quality

The relationship between lice infestation in calves during their first winter and damage to the leather produced was investigated in a trial involving 500 calves, 100 of which were treated with a pour-on endectocide during the first winter. All the calves received routine lice treatment in the second winter and were reared to slaughter weight. The hides were removed at the abattoir, tanned, inspected for lice-related damage, and graded according to their suitability for the production of high quality leather. The untreated group developed natural infestations of the chewing louse (Bovicola bovis) during the first winter but none was observed in the second winter. Hides from cattle infested with lice in their first winter had higher levels of lice damage than hides from those treated with eprinomectin, at both the chrome-tanned and dried dyed-crust stages of leather production.     

The efficacy of injectable and pour-on formulations of moxidectin against lice on cattle

SUMMARY The efficacy of one administration of moxidectin against natural infestations of the 3 common species of cattle lice in Australia: Linognathus vitull, Damallnla bovls and Haematoplnus eurysternus, was determined. A high degree of control of L vituli (too long-nosed sucking louse) was achieved with both the cattle Injection (96.7% and 100%) at 0.2 mg/kg and the cattle pour-on formulation (94.6% and 100%) at 0.5 mg/kg on the 2 farms where this louse occurred. An Injection of moxidectin at 0.2 mg/kg did not eliminate D bovis. In these trials efficacy ranged from nll to 85.2%. In contrast the pour-on formulation at 0.5 mg/kg provided consistently high efficacy (83.6 to 100%) against D bovis on 3 farms. Infestations of H eurysternus (the short-nosed sucking louse) were low and no significant difference was recorded between treatment groups In a single trial, however, lice persisted on 3 to 7 cattle In the untreated group at each Inspection, whereas none were found on any animals in the 2 groups treated with moxidectin on Inspection after treatment.     

Eradication of lice in cattle.

The purpose of this field study was to develop and evaluate eradication as a strategy to control lice in cattle. Thirty-three herds of cattle were selected and observed during a period of two and a half years. Before eradication, biting lice (Damalinia bovis) were present in 94% of the herds and 27% of the animals. Sucking lice (Linognathus vituli) were present in 42% of the herds and 5% of the animals. These levels were very similar to those reported from other countries in Northern Europe. The eradication strategy was successful in 28 of 33 herds, but lice were still present in 5 herds 3 to 6 months after treatment. Biting lice were present in all these 5 herds, sucking lice were present in 3 herds. During the next 12 months, nine of the 28 herds were reinfected with lice. Six herds were reinfected with just biting lice, 2 herds with just sucking lice and one herd was reinfected with both. There was no significant difference between the 2 louse species regarding the risk of unsuccessful eradication or reinfection. The only significant risk factor for reinfection was either purchase of livestock or use of common pasture, combined with failure in pre-treatment of newly introduced animals.     

EFFECTS OF ARTIFICIALLY INDUCED INFESTATIONS OF THE CATTLE LOUSE, LINOGNATHUS VITULI

SUMMARY Groups of White Shorthorn calves, stanchioned to prevent self-grooming and given high or low levels of nutrition, were exposed to artificial infestations of the cattle louse, Linognathus vituli. Undernutrition caused a significant (P = 0.03) increase in lice numbers. Lice did not significantly affect percentages of packed cells, lymphocytes, polymorphs and eosinophils or the number of red and white cells. Calves were always able to control lice infestations without self-grooming.     

Comparative efficacy of several insecticides for control of cattle lice (Mallophaga: Trichodectidae and Anoplura: Haematopinidae)

Several insecticides were applied to cattle at various rates, mixtures, application methods and numbers of treatments and evaluated for control of cattle lice. The insecticides included permethrin at various rates as a pour-on with and without the synergist piperonyl butoxide (PBO), and also at various rates as a spray with and without PBO. Pirimiphos-methyl was evaluated as a pour-on at various rates and with one or two applications. Lambda cyhalothrin in microencapsulated formulation was compared to a non-microencapsulated formulation. Rabon(R) was applied as a pour-on at two rates and at one or two treatments for each rate. Dimilin(R) was applied as a pour-on at two rates and mixed with permethrin and applied at two rates. Five endectocides, Eprinex(R), Ivomec(R), Dectomax(R), Cydectin(R) and Phoenectin(R) were all applied at the same rate. The cattle utilized in this research were all infested with a mixture of lice species. The species included: Bovicola (Damalinia) bovis (L.), Haematopinus eurysternus (Nitzsch), Linognthus vituli (L.) and Solenoptes capillatus (Enderlein). Most of the cattle were infested with at least two species and some had three or all four species present. All of the treatments except permethrin 1.0%+5% PBO, at a rate of 3ml/45.4kgwt. provided acceptable lice control with one application. Data indicated that applying the insecticides in early January should provide enough control to render the need for a second treatment unnecessary.     

Survey on blood-sucking lice (Phthiraptera: Anoplura) of ruminants and pigs with molecular detection of Anaplasma and Rickettsia spp

Lice may serve as biological or mechanical vectors for various infectious agents. To investigate louse infestation of ruminants and pigs, and pathogens potentially transmitted by them, anopluran lice (n=1182) were collected in Hungary, and evaluated for the presence of anaplasma, rickettsia and haemotropic mycoplasma DNA. On cattle the following species were found: Linognathus vituli (57%), Haematopinus eurysternus (38%) and Solenopotes capillatus (5%). L. vituli had a lower mean individual count/host when compared to H. eurysternus. On calves only L. vituli was observed, with a higher louse burden than on full-grown cattle. H. eurysternus and S. capillatus were more likely to occur simultaneously with another species on the same host, than L. vituli. Goats infested with Linognathus stenopsis had the overall highest prevalence (68%), while pigs harbouring Haematopinus suis showed the lowest (<1%). Anaplasma DNA was detected in 50% of pools analysed. In L. vituli Anaplasma ovis (or a closely related novel Anaplasma marginale genotype) was identified. Anaplasma-positivity of H. suis suggests that pigs may extend the reservoir and/or host spectrum of relevant species. Anaplasma-infected L. stenopsis pools show for the first time that caprine anaplasmosis is endemic in Hungary. Rickettsia spp. were demonstrated from Linognathus spp. and H. eurysternus. No haemotropic mycoplasmas were detected in any samples. In conclusion, this is the first molecularly confirmed report of bovine and ovine Anaplasma spp. in L. vituli, L. stenopsis and H. suis. The present results suggest that phthirapterosis of domestic animals deserves more attention, and lice should be evaluated among the broad range of potential vectors of arthropod-borne pathogens.     

The effect of simulated rainfall on the efficacy of doramectin pour-on against nematode parasites of cattle.

Two studies were conducted with doramectin topically administered at 500 microg/kg body weight to assess retention of therapeutic efficacy against nematode infections of cattle before, and after, simulated rainfall. In the first study, 50 heifers, with patent nematode infections, were allocated to one of five treatment groups. An untreated control group and one doramectin-treated group were not exposed to simulated rainfall. Simulated rainfall was applied at a rate of 25.4 mm of water in 35 min to three of the five groups: one group immediately before treatment, the second group 90 min after treatment, and the third group 24 h after treatment. Fecal samples were collected for determining egg counts 14 days after treatment. Percentage efficacy ranged from 97.3% to 100% in all treated calves, regardless of exposure to simulated rainfall. The second study involved 40 mixed-sex cattle that were allocated to one of four treatment groups (one saline control and three doramectin-treated groups). All cattle were confirmed to be free of nematode infections prior to oral gavage with infective larvae of Dictyocaulus viviparus, Cooperia oncophora, and Ostertagia ostertagi. Twenty-six days after infection, three groups were treated with doramectin pour-on and exposed to 20 mm of simulated rainfall over 40 min: one group 60 min before treatment, the second 20 min after treatment, and the third 40 min after treatment. Approximately two weeks after treatment, all cattle were necropsied for worm counts. In all treated groups, the percentage efficacy against O. ostertagi and D. viviparus was >99% to 100%. Percentage efficacy against Cooperia ranged from 97% to 98%. Results indicated that doramectin pour-on remains efficacious against nematodes of cattle when administered immediately before or after rainfall.     

Arthropod parasites of Nubian ibexes (Capra ibex nubiana) and gazelles (Gazella gazella) in Israel.

In a 20-year survey the following ectoparasites were collected from Nubian ibexes: larvae of an unidentified Oestrus sp. collected from the nasal cavities, sinuses and horns, hippoboscid flies (Lipoptena chalcomelaena) specific to the Nubian ibex, blood sucking lice (Linognathus africanus) and unidentified biting lice (Damalinia sp.). Ibexes were severely infested with the cattle tick, Boophilus annulatus; a few Hyalomma anatolicum excavatum ticks were also collected. In five ibexes kept in two zoos, showing otitis, Psoroptes cuniculi, was identified, and from skin scrpaings of nine animals with severe dermatitis in three other zoos, Sarcoptes scabiei was isolated. Infestation of Nubian ibexes with sucking and biting lice as well as H. anatolicum excavatum is reported for the first time. The ectoparasites collected from gazelles were: hippoboscid flies (Lipoptena capreoli), calliphorid flies (Lucilia sericata and Calliphora sp.), sucking lice (Linognathus africanus and Solenopotes capillatus) and unidentified biting lice (Damalinia sp.), fleas (Ctenocephalides felis felis), and ticks, B. annulatus, Rhipicepahlus bursa, Rhipicephalus turanicus, H. anatolicum excavatum and H. marginatum rufipes. In skin scrapings of four gazelles with local dermatitis in the fetlocks Chorioptes bovis was identified. Neoschoengastia sp. was found in craters between the claws in three gazelles.     

The effectiveness of a single dose of doramectin pour-on in the control of gastrointestinal nematodes in yearling stocker cattle

Three field studies were conducted to determine the efficacy of a single dose of doramectin pour-on in the control of gastrointestinal nematode infections in yearling stocker calves on pasture. These 140-day studies were carried out between October 1995 and March 1996 in Tennessee (TN), between January and June 1997 in Louisiana (LA), and between May and September 1997 in Wisconsin (WI). Calves with patent nematode infections were equally allocated to treatments (doramectin pour-on, at 500 microg/kg body weight or untreated control) and pastures as randomized complete-block designs (LA and TN studies) or completely at random (WI study). There were six pasture replicates per treatment at each site, with each pasture replicate accommodating six calves at the TN site (36 calves per treatment), five calves at the LA site (30 calves per treatment), and seven calves at the WI site (42 calves per treatment). Fecal samples for nematode egg counts were collected on Day 0, and at 28-day intervals thereafter. Body weights were recorded on Day 0 and at 28-day intervals until study termination. Nematode egg output of the doramectin-treated groups was reduced over the entire grazing period compared to those in the untreated control groups, resulting in average daily weight-gain advantages of 0.055 kg (p < or = 0.05) for the TN study, 0.208 kg (p < or = 0.05) for the LA study, and 0.116 kg (p < or = 0.05) for the WI study.     

Propetamphos pour-on formulation for the control of lice on sheep: effect of lice on weight gain and wool production

Infestation of sheep with the biting louse Damalinia ovis is a potentially serious problem which can effect the quality and quantity of wool produced and may reduce weight gains. In this trial treated sheep produced 34 per cent more wool than louse-infested controls and the wool from the treated sheep was of a better quality. The mean liveweight gain in the treated groups was 18 per cent more than that in the untreated group. Treatment with a pour-on formulation of propetamphos at 25 to 50 mg kg-1 gave more than 99 per cent control of lice and protected against reinfestation for four months.     

EFFICIENCY AND SAFETY OF METHIDATHION APPLIED AS A POUR-ON SYSTEMIC INSECTICIDE FOR CONTROL OF CATTLE LICE

Methidathion applied to cattle as a pour-on insecticide for control of lice (predominantly Linognathus vituli, but also Haematopinus eurysternus and Damalinia bovis) caused a reduction in the lice population of approximately 98% to 99% in laboratory trials at the minimum recommended dose of 4 mg/kg and a 98.8% to 100% efficiency was achieved in field trials. In a comparative efficiency trial in the laboratory methidathion at 3.5 to 5 mg/kg reduced the lice population by 98.8% to 99%, fenthion by 98.5% at 4.5 mg/kg and famphur by 99.7% at 16.5 mg/kg. Methidathion was tolerated by calves aged 15 to 20 weeks at dose rates up to 40 mg/kg indicating an approximate 7 fold safety margin, but 1 of 4 calves treated at 50 mg/kg died following treatment. Treatment with fenthion at 50 mg/kg, 7.4 times the average recommended rate, famphur at 75 mg/kg, 3 times the average rate and chlorpyrifos at 85 mg/kg, 5 times average rate, caused reductions in whole blood cholinesterase activity of 52%, 27% and 47% respectively which were similar to the reductions in cholinesterase activity found in calves treated with methidathion at similar levels above the recommended commercial dose rates. It was found that 2 day old calves were more sensitive to treatment with methidathion than calves 9 or 16 days old. A further 11,900 cattle of varying age, breed and sex were treated with methidathion under field conditions at the recommended rate of 4 to 8 mg/kg, and 534 cattle were treated at 24 mg/kg without any signs of toxicity.