Normal temperature variation in New Zealand white rabbits during restraint for preliminary pyrogen test.

The in vivo pyrogen test is the main toxicological assay used in the quality control of injectable products, especially immunobiologicals. Pharmacopoeias state that, before the main test, a preliminary test must be conducted on all animals, and must follow the same conditions as the main test. The aim of this study was to determine the normal temperature variation in New Zealand white rabbits during restraint and propose a limit value for considering an animal as suitable for testing. Results of the temperature variation in 4,689 rabbits during preliminary tests were obtained from the routine database of the Pharmacology and Toxicology Department of the National Institute of Quality Control in Health (INCQS/FIOCRUZ), Brazil. From these preliminary tests, 3,364 rabbits were considered suitable for testing according to the Brazilian Pharmacopoeia criteria (temperature variation < 0.5 °C). Results showed that about 92 per cent of the rabbits presented a normal individual temperature variation equal to or below 0.30 °C. Animals presenting a temperature variation close to the fever temperature must not be included in the main test, since they can be stressed or sick. Consequently, the temperature variation of 0.30 °C could be adopted by pharmacopoeias as a limit temperature to be considered in the preliminary test to determine which animals can be used in the main rabbit pyrogen test. Animals can be pre-tested until presenting this safe variation, in order to ensure they are healthy and minimise interference in the result.

[Influence of temperature and humidity on the nymphal development of Rhodnius robustus]. / Influência da temperatura e umidade no desenvolvimento ninfal de Rhodnius robustus.

OBJECTIVE: To assess the significant differences in the nymphal development of the Rhodnius robustus Larrousse, 1927 under different temperatures and humidity conditions. This is a species found in the northern region of Brazil (states of Acre, Amazonas, and Pará), Colombia, Equator, Peru and Venezuela. METHODS: Three groups of triatominae were kept under the following laboratory conditions: 33/40 (33+1 degree C and 40+/-5% of relative humidity - RH), 33/70 (33+/-1 degree C and 70+/-5% RH), and 28/70 (28+/-1 degree C and 70+/-5% RH). The incubation period of the eggs, developmental time of each stage, mortality percentage, number of bloodmeals, and the total amount of time from the egg hatching to adult ecdysis were observed. RESULTS: The shortest average time of nymphal development was observed in the 28/70 group, with following averages: 14.4, 17.3, 20.3, 22.8, and 40. Significant differences were observed in the embryonic development between the groups (p<0.01). For all groups, the number of bloodmeals had a gradual increase near the adult phase, except for the 3rd instar of the 33/70 group. The smallest mortality percentages were seen in the 28/70 group. CONCLUSIONS: High temperatures, regardless of the humidity, can accelerate the biological development cycle of R. robustus. However, these temperatures can impair the colonies' survival, preventing their maintenance in the laboratory.

Biodistribution study of the anaesthetic sodium phenobarbital labelled with technetium-99m in Swiss mice infected with Schistosoma mansoni Sambon, 1907.

Technetium-99m (99m Tc) is a radionuclide that has negligible environmental impact, is easily available, inexpensive and can be used as a radioactive tracer in biological experiences. In order to know the mode of action of sodium phenobarbital in moving adult Schistosoma mansoni worms from mesenteric veins to the liver, we labelled sodium phenobarbital (PBBT) with 99mTc and a biodistribution study in infected and non-infected Swiss mice was performed. The PBBT was incubated with stannous chloride used as reducing agent and with 99mTc, as sodium pertechnetate. The radioactivity labelling (%) was determined by paper ascending chromatography performed with acetone (solvent). The 99mTc-PBBT was administered by intraperitoneal route to Swiss mice infected eight weeks before. The animals were perfused after different periods of time (0,1,2,3,4 hr) when blood, spleen, liver, portal vein, mesenteric veins, stomach, kidneys and adult worms were isolated. The radioactivity present in these samples was counted in a well counter and the percentage was determined. The radioactivity was mainly taken up by the blood, kidney, liver and spleen. No radioactivity was found on the adult worms. We concluded that the worm shift was due to an action on the host of the sodium phenobarbital.

Schistosoma mansoni Sambon, 1907: comparative morphological studies of some Brazilian strains.

The morphology of Schistosoma mansoni adult male worms from three strains which have been maintained in albino mice for several generations, was compared to a strain that has been isolated from the natural host Nectomys squamipes (Rodentia:Muridae) captured in Sumidouro (Rio de Janeiro State) and have been maintained in the same sylvatic rodent under laboratory conditions. Total length of specimens, distance between suckers, the number of testes and extention of testes grouping were the taxonomic characters analysed. The worms recovered from N. squamipes showed expressive differences (p < 0.01) compared to the other strains regarding the considered morphological characters. The strains that were maintained in mice presented statistical differences (p < 0.01) in several characters. Some adult worms besides the normal position of the testes also showed an atypical arrangement of these glands. It can be concluded that the morphology of adult worms may be used to distinguish S. mansoni strains and that morphological changes in adult worms are not induced by successive inoculations of a strain in mice.

Host-induced morphological changes of Schistosoma mansoni Sambon, 1907 male worms.

In order to evaluate the permissiveness of Nectomys squamipes to Schistosoma mansoni and the influence of the albino mice on the morphological aspects of adult worms derived from a population isolated from N. squamipes, the morphology of adult S. mansoni Sambon, 1907 male worms was studied using a digital image analyser (MOP VIDEOPLAN) and light microscopy. Their sources were as follows: (1) recovered from the wild rodent N. squamipes Brants naturally infected from Sumidouro, RJ, Brazil; (2) recovered from albino mice experimentally infected with the strain derived from N. squamipes; (3) recovered after the isolation of a strain derived from aboriginal human infections in Sumidouro. Worms recovered from N. squamipes (group 1) showed body length and distance between suckers significantly bigger than those of the specimens maintained in mice (groups 2 and 3). The number of testes in group 1 was statistically less than that of groups 2 and 3. Group 2 strains which were maintained in mice, presented the length of the worms as the only significant different character. Data show that: (1) N. squamipes is a more suitable host for the development of S. mansoni when compared to the albino mice; (2) a strain of S. mansoni isolated from a natural host undergoes morphological changes after its passage in the white mouse.

Comparative study on the localization of adult Schistosoma mansoni worms in albino mice anesthetized with pentobarbital sodium, ether or chloroform.

The effect of anesthetic drugs on the localization of adult worms in albino mice was compared. The animals with 56 days of infection were anesthetized with pentobarbital sodium, ether or chloroform. Perfusion was carried out immediately after, recovering the worms and classifying them in relation to their localization on the liver or portal vein and the mesenteric veins. Our results showed that pentobarbital sodium produced a greater displacement of the worms to the liver (89%) than ether (76%) and chloroform (34%) did, when compared to the control group (22%). The difference between pentobarbital sodium and ether was significant (p < 0.05). We suggest that anesthetic drugs may not be used in studies on the distribution of adult worms in several hosts.

Effect of beta-myrcene on pentobarbital sleeping time.

Evidence that beta-myrcene (MYR) interferes with the metabolic activation of premutagens has been provided by in vitro studies. In order to determine whether MYR also interferes with the in vivo metabolism of xenobiotics, thereby modifying pharmacological responses to drugs, we investigated the effects of this monoterpene on pentobarbital (PT) sleeping time in rats. Two experiments were carried out. In the first, a single dose of MYR (0.25, 0.5 or 1.0 g/kg po) was given 1 h before PT (40 mg/kg ip). No effect was observed with the two lowest doses, but the highest MYR dose given 1 h before PT increased the PT-induced sleeping time (131 +/- 15 min vs 64 +/- 15 min for controls, mean +/- SD). In the second experiment, male rats were treated with MYR (1.0 g/kg po once a day) for 14 days and injected with PT (40 mg/kg ip) 24 h after the last dose of MYR. Repeated treatment with MYR markedly reduced PT sleeping time compared to the vehicle-treated control group (21 +/- 13 min vs 35 +/- 19 min for controls, mean +/- SD). These results indicate that MYR interferes with the in vivo barbiturate metabolism and support the view that MYR induces the phenobarbital-inducible cytochrome P-450 (P-450 2B subfamily) enzymes in the rat.

Effect of beta-myrcene on pentobarbital sleeping time

Evidence that beta-myrcene (MYR) interferes with the metabolic activation of premutagens has been provided by in vitro studies. In order to determine whether MYR also interferes with the in vivo metabolism of xenobiotics, thereby modifying pharmacological responses to drugs, we investigated the effects of this monoterpene on pentobarbital (PT) sleeping time in rats. Two experiments were carried out. In the first, a single dose of MYR (0.25, 0.5 or 1.0 g/kg po) was given 1 h before PT (40 mg/kg ip). No effect was observed with the two lowest doses, but the highest MYR dose given 1 h before PT increased the PT-induced sleeping time (131 +/- 15 min vs 64 +/- 15 min for controls, mean +/- SD). In the second experiment, male rats were treated with MYR (1.0 g/kg po once a day) for 14 days and injected with PT (40 mg/kg ip) 24 h after the last dose of MYR. Repeated treatment with MYR markedly reduced PT sleeping time compared to the vehicle-treated control group (21 +/- 13 min vs 35 +/- 19 min for controls, mean +/- SD). These results indicate that MYR interferes with the in vivo barbiturate metabolism and support the view that MYR induces the phenobarbital-inducible cytochrome P-450 (P-450 2B subfamily) enzymes in the rat

Neurobehavioral study of the effect of beta-myrcene on rodents.

Tea prepared from lemongrass (Cymbopogon citratus) is used for its supposed anxiolytic, hypnotic and analgesic properties in Brazilian folk medicine. beta-Myrcene, a major constituent of lemongrass, produces analgesia in rodents but there is some controversy about whether this action is central or peripheral or both. Rats and mice received beta-myrcene, 1 g/kg po in corn oil, or corn oil alone 1 h before being evaluated for a series of responses which included exploratory and emotional behavior, anxiolytic activity in a plus maze and inhibition of conditioned avoidance. No evidence was demonstrable for an effect of beta-myrcene on any of these behaviors. Similarly, beta-myrcene had no protective effect on pentylenetetrazol (PTZ)-induced seizures in mice. These data suggest that beta-myrcene has no benzodiazepine-like anxiolytic activity and that an activity on the central nervous system (antidepressive or antipsychotic) is unlikely. Despite the negative results of this study, folk use of lemongrass tea may still be justified by its analgesic properties.

Toxicological study of the molluscicidal latex of Euphorbia splendens: irritant action on skin and eye.

Aqueous solutions of the molluscicidal latex of Euphorbia splendens are irritant to the rabbit eye in concentrations higher than 0.35% and to the rabbit skin in concentrations higher than 0.5%. Although this irritant potential does not preclude its use as a molluscicide, special precautions are recommended for handling and application of the product and the hazard of skin tumor-promoting potential should be carefully investigated before its use for schistosomiasis vector control.

Toxicological study of the molluscicidal latex of Euphorbia splendens: irritant action on skin and eye

Aqueous solutions of the molluscicidal latex of Euphorbia splendens are irritant to the rabbit eye in concentrations higher than 0.35% and to the rabbit skin in concentrations higher than 0.5%. Although this irritant potential does not proclude its use as a molluscicide, special precautions are recommended for hanbdling and application of the product and the hazard of skin tumor-promoting potencial should be carefully investigated before its use for schistosomiasis vector control.

Neurobehavioral study of the effect of beta-myrcene on rodents

Tea prepared from lemongrass (Cymbopogon citratus) is used for its supposed anxiolytic, hypnotic and analgesic properties in Brazilian folk medicine. beta-Myrcene, a major constituent of lemongrass, produces analgesia in rodents but there is some controversy about whether this actions is central or peripheral or both. Rats and mice received beta-myrcene, 1 g/Kg po in corn oil alone 1 h before being evaluated for a series of responses which included exploratory and emotional behavior, anxiolytic activity in a plus maze and inhibition of conditioned avoidance. No evidence was demonstrable for an effect of beta-myrcene on any f these behaviors. Similarly, beta-myrcene had no protective effect on pentylenetetrazol (PTZ)-induced seizures in mice. These data suggest that beta-myrcene has no benzodiazepine-like anxiolytic activity and that an activity on the central nervous system (antidepressive or antipsychotic) is unlikely. Despite the negative results of this study, folk use of lemongrass tea may still be justified by its analgesic properties

Comparison of five methods for the determination of lethal dose in acute toxicity studies.

The aim of the present study was to compare the reliability of LD50 determination using the traditional Litchfield and Wilcoxon method with that obtained by four alternative tests requiring smaller numbers of animals, for the purpose of classifying chemicals according to their acute toxicity. Acute lethal dose determinations were carried out in mice for oral and intraperitoneal administration of hexachlorophene, lidocaine, methanol, phenobarbital and physostigmine. The Molinengo method proved not to be as reliable as suggested by its author. Determination of LD50 using the Thompson and Weil method or, alternatively, the maximal non-lethal dose and the approximate lethal dose permitted the classification of the chemicals in essentially the same order. The approximate lethal dose method, in particular, seems to be a very suitable alternative method to the classical LD50 test since it requires only about 6 animals, provides enough information to order chemicals according to their toxicities, and provides useful information for planning subsequent repeated-dose studies.

Comparison of five methods for the determination of lethal dose in acute toxicity studies

The aim of the prsent study was to compare the realibility of LD50 determination using the traditional Litchfield and Wilcoxon method with that obtained by forur alternative tests requiring smaller numbers of animals, for the purpose of classifyng chemicals according to their acute toxicity. Acute lethal dose determinations were carried out in mice for oral and intraperitoneal administration of hexachlorophene, lidocaine, methanol, phenobarbital and physostigmine. The Molinengo method proved not to be as reliable as suggested by its author. Determination of LD50 using the Thompson and Weil method or, alternatively, the maximal non-lethal dose and the approximate lethal dose permitted the classification of the chemicals in essentially the same order. The approximate lethal dose method, in particular, seems to be a very suitable alternative method to the classical LD50 test since it requires only about 6 animals, provides enough information to order chemicals according to their toxicities, and provides useful information for planning subsequent repeated-dose studies