Relationship of arsenic and chromium availability with carbon functional groups, aluminum and iron in Little Washita River Experimental Watershed Reservoirs, Oklahoma, USA.

Sediment from three reservoirs located in the Little Washita River Experimental Watershed (LWREW) in Oklahoma, USA with contrasting dominant land uses were analyzed for total and extractable concentrations of arsenic (As) and chromium (Cr), and the potential ecologic risk to benthic organisms. Extractable As ranged from 0.24 to 1.21 mg kg-1, in the order grazing>cropland>forest and 0.13-0.58 mg kg-1 for extractable Cr, in the order of forest>grazing>cropland. However, only approximately < 1.5% of total As and < 4% of total Cr were extractable. Total As ranged from 16.2 to 141 mg kg-1 and total Cr ranged from 5.06 to 40.1 mg kg-1 both in the order of cropland>grazing>forest. The sediment exhibited an alkaline pH (8.0-8.7). As sorption exhibited a positive relationship with Al (r = 0.9995; P = 0.0001), Fe (r = 0.9829; P = 0.0001), and C (r = 0.4090; P = 0.0017) and Cr correlated positively with Al (r = 0.9676 P = 0.0001), Fe (r = 0.9818; P = 0.0001), and C (r = 0.3368; P = 0.0111). In addition, both As and Cr exhibited positive relationships with carbon (C) functional groups in the order of O-alkyl C> methoxyl C> alkyl C> aromatic C> carboxyl C> phenolic C. The sediment concentration analysis results illustrated that As in all reservoirs exceeded their respective Threshold Effect Level (TEL) and/or Probable Effect Level (PEL) indicating that existing concentrations of metals in these sediments were sufficiently high to cause adverse effects. However, Cr concentrations in all reservoirs evaluated was lower compared to the TEL and PEL.

Fusion of Na-ASP-2 with human immunoglobulin Fcγ abrogates histamine release from basophils sensitized with anti-Na-ASP-2 IgE.

Na-ASP-2 is a major protein secreted by infective third-stage larvae (L3) of the human hookworm Necator americanus upon host entry. It was chosen as a lead vaccine candidate for its ability to elicit protective immune responses. However, clinical development of this antigen as a recombinant vaccine was halted because it caused allergic reactions among some of human volunteers previously infected with N. americanus. To prevent IgE-mediated allergic reactions induced by Na-ASP-2 but keep its immunogenicity as a vaccine antigen, we designed and tested a genetically engineered fusion protein, Fcγ/Na-ASP-2, composed of full-length Na-ASP-2 and truncated human IgG Fcγ1 that targets the negative signalling receptor FcγRIIb expressed on pro-allergic cells. The chimeric recombinant Fcγ/Na-ASP-2 protein was expressed in Pichia pastoris and shared the similar antigenicity as native Na-ASP-2. Compared to Na-ASP-2, the chimeric fusion protein efficiently reduced the release of histamine in human basophils sensitized with anti-Na-ASP-2 IgE obtained from individuals living in a hookworm-endemic area. In dogs infected with canine hookworm, Fcγ/Na-ASP-2 resulted in significantly reduced immediate-type skin reactivity when injected intradermally compared with Na-ASP-2. Hamsters vaccinated with Fcγ/Na-ASP-2 formulated with Alhydrogel(®) produced specific IgG that recognized Na-ASP-2 and elicited similar protection level against N. americanus L3 challenge as native Na-ASP-2.

Tumorigenicity of sodium ascorbate in male rats.

Sodium ascorbate, like other sodium salts such as saccharin, glutamate, and bicarbonate, produces urinary alterations when fed at high doses to rats, which results in mild superficial urothelial cytotoxicity and regeneration but not tumors in a standard 2-year bioassay. Sodium saccharin was shown to produce a low incidence of bladder tumors in rats if administered in a two-generation bioassay. In the present study, we evaluated sodium ascorbate in a two-generation bioassay that involved feeding to the male and female parental F344 rats for 4 weeks before mating, feeding the dams during gestation and lactation, and then feeding the weaned (at 28 days of age) male F1 generation rats for the remainder of their lifetime (up to 128 weeks of the experiment). Dietary levels of 1.0, 5.0, and 7.0% sodium ascorbate were tested. At 5.0 and 7.0% sodium ascorbate, there was an increase in urinary bladder urothelial papillary and nodular hyperplasia and the induction of a few papillomas and carcinomas. There was a dose-responsive increase in renal pelvic calcification and hyperplasia and inhibition of the aging nephropathy of rats even at the level of 1% sodium ascorbate. Because the short-term urothelial effects of sodium ascorbate in rats are inhibited by treatments producing urinary acidification to pH < 6.0, we coadministered high doses of long-term NH4Cl to groups of rats with 5.0 or 7.0% sodium ascorbate to evaluate the long-term effects. The combination of 7.0% sodium ascorbate plus 2.78% NH4Cl in the diet was toxic, and the group was terminated early during the course of the experiment. The group fed 5.0% sodium ascorbate plus 2.04% NH4Cl showed complete inhibition of the urothelial effects of sodium ascorbate and significant inhibition of its renal effects. We also demonstrated the presence of a calcium phosphate-containing urinary precipitate in rats fed sodium ascorbate at all doses, in a dose-responsive manner. The formation of the precipitate was inhibited by coadministration with NH4Cl. The proliferative effects of sodium ascorbate on the male rat urinary tract in this study are similar to those seen with sodium saccharin when administered in a two-generation bioassay. Mechanistic information suggests that this is a high-dose, rat-specific phenomenon.

Potential arsenic exposures in 25 species of zoo animals living in CCA-wood enclosures.

Animal enclosures are often constructed from wood treated with the pesticide chromated copper arsenate (CCA), which leaches arsenic (As) into adjacent soil during normal weathering. This study evaluated potential pathways of As exposure in 25 species of zoo animals living in CCA-wood enclosures. We analyzed As speciation in complete animal foods, dislodgeable As from CCA-wood, and As levels in enclosure soils, as well as As levels in biomarkers of 9 species of crocodilians (eggs), 4 species of birds (feathers), 1 primate species (hair), and 1 porcupine species (quills). Elevated soil As in samples from 17 enclosures was observed at 1.0-110mg/kg, and enclosures housing threatened and endangered species had As levels higher than USEPA's risk-based Eco-SSL for birds and mammals of 43 and 46mg/kg. Wipe samples of CCA-wood on which primates sit had dislodgeable As residues of 4.6-111µg/100cm(2), typical of unsealed CCA-wood. Inorganic As doses from animal foods were estimated at 0.22-7.8µg/kg bw/d. Some As levels in bird feathers and crocodilian eggs were higher than prior studies on wild species. However, hair from marmosets had 6.37mg/kg As, 30-fold greater than the reference value, possibly due to their inability to methylate inorganic As. Our data suggested that elevated As in soils and dislodgeable As from CCA-wood could be important sources of As exposure for zoo animals.

Cleaning-induced arsenic mobilization and chromium oxidation from CCA-wood deck: Potential risk to children.

Concern about children's exposure to arsenic (As) from wood treated with chromated-copper-arsenate (CCA) led to its withdrawal from residential use in 2004. However, due to its effectiveness, millions of American homes still have CCA-wood decks on which children play. This study evaluated the effects of three deck-cleaning methods on formation of dislodgeable As and hexavalent chromium (CrVI) on CCA-wood surfaces and in leachate. Initial wipes from CCA-wood wetted with water showed 3-4 times more dislodgeable As than on dry wood. After cleaning with a bleach solution, 9.8-40.3µg/100cm(2) of CrVI was found on the wood surface, with up to 170µg/L CrVI in the leachate. Depending on the cleaning method, 699-2473mg of As would be released into the environment from cleaning a 18.6-m(2)-deck. Estimated As doses in children aged 1-6 after 1h of playing on a wet CCA-wood deck were 0.25-0.41µg/kg. This is the first study to identify increased dislodgeable As on wet CCA-wood and to evaluate dislodgeable CrVI after bleach application. Our data suggest that As and CrVI in 25-year old CCA-wood still show exposure risks for children and potential for soil contamination.

Early life protein malnutrition changes exploration of the elevated plus-maze and reactivity to anxiolytics.

In order to investigate whether protein malnutrition in early life causes lasting changes in reactivity to anxiolytic drugs, exploration of the elevated plus-maze was used. Rat dams during lactation (21 days) and pups after weaning until day 49 of life were fed on 8% casein diet (M rats), while their well-nourished controls received 25% casein (W rats). From day 50 on all animals ate the same balanced diet. Experiments started on day 70. Under the non-drug condition, M rats tended to explore the open arms of the maze relatively more than W rats. Diazepam (0.5-5 mg/kg, IP) dose-dependently increased the percentage of open/total arm entries without significantly affecting the total number of arm entries in W rats. This selective anxiolytic effect of diazepam was considerably smaller in M rats. Ipsapirone (0.5-5 mg/kg) caused a similar though less pronounced anxiolytic effect in W rats, whereas the drug decreased both the % open/total and total arm entries in M rats. In contrast, ritanserin (0.05-1 mg/kg) significantly increased the % open/total arm entries in M rats only, though not in a dose-dependent way. Isamoltane (2.5-20 mg/kg) was ineffective on both M and W rats. These results indicate that early protein malnutrition causes long-lasting alterations in brain systems regulating emotional behaviour.

Blockade of a motor response by cholinergic stimulation of the toad midbrain tegmentum.

Application of an acid solution to the dorsal skin of conscious toads having intact nervous system induces a scratching reflex and escape movements, as well as autonomic alterations (hypertension and tachycardia) that are part of the defense response. The motor components of this response are abolished or reduced by microinjection of 60, 30, 15 or 7.5 ng carbachol into the midbrain tegmentum. The cardiovascular components, however, continue to be present, although their amplitude is reduced. The depression of the motor response is statistically significant up to 15 minutes for the 60 ng dose, up to 10 minutes for the 15 and 30 ng doses, and only up to 5 minutes for the 7.5 ng dose. The data suggest that the midbrain tegmentum may modulate the reflex motor response triggered by a noxious stimulus and also participate in the organization of the escape movements. The importance of cholinergic agents in this modulation is discussed. The persistence of the cardiovascular component of the response shows the importance of this parameter as an indicator of alert situations.

Changes in electric activity (EEG) of the telencephalon of conscious toads (Bufo paracnemis) caused by cholinergic stimulation of the mesencephalic tegmentum.

Studies were carried out to investigate the effect of microinjection of cholinergic agonist carbamylcholine (carbachol) into the mesencephalic tegmentum of conscious toads (Bufo paracnemis), subjected or not to section of the olfactory nerves, on the electrical activity (EEG) of the telencephalon. The response depended on the type of preexisting EEG activity. Marked desynchronization was observed both in animals with intact nerves and in animals with sectioned nerves, whose previous basal activity was already desynchronized (high frequency and low amplitude). In toads with synchronized basal activity and with intact nerves, an increase, a fall or even abolition of preexisting spindle-like activity occurred, whereas in animals with sectioned nerves the effect was reduction or abolition of spindle-like activity. The data show that stimulation of the mesencephalic tegmentum can alter the electrical activity of the telencephalon and that this effect is not only secondary to a change in nasal air flow alterations since it occurs in toads with sectioned olfactory nerves.

Effects of malnutrition and environment on the acquisition and extinction of avoidance behavior in rats.

Twelve newborn rats were fed by mothers maintained on a protein-deficient diet (12% casein, M) during lactation, and 12 rats fed by mothers maintained on a diet containing 25% casein were used as controls (C). After weaning, all animals were fed standard lab ration. Half of each group was housed individually (MI and CI), while the other half was allowed to live in pairs (MP and CP). When adult, all animals were trained to avoid footshocks by jumping onto a platform. Training sessions consisted of 40 trials starting with a 20 sec light stimulus (CS) and followed by a 2 sec, 0.6 mA shock (US) with an average intertrial interval of 54 sec. When all animals displayed consistent avoidance behavior, the extinction phase was initiated. The procedure was the same as for the training sessions except that the shock generator was disconnected. Extinction continued until each animal showed a 50% reduction in avoidance performance. During acquisition, MI learned faster than CI and CI showed greater avoidance performance than CP, but no differences were observed between MP and CP. During extinction, group M responded more persistently than group C. The present acquisition results may explain the contradictory data reported in the literature with respect to the effects of malnutrition on avoidance performance, since environmental stimulation was shown to reduce the effects of early malnutrition. Individually housed animals showed greater avoidance performance during both phases.

Effects of early protein malnutrition and repeated testing upon locomotor and exploratory behaviors in the elevated plus-maze.

An elevated plus-maze was used to investigate the effects of repeated testing on the locomotor and exploratory behaviors of malnourished rats. Pup malnutrition was induced during the lactation period (0 to 21 days of age) by feeding the dams a protein-deficient diet (6% protein) and the animals were allowed to recover from weaning to 70 days of age by eating a commercial lab chow diet. Control animals were suckled by dams receiving a normal protein diet (16% protein) during the lactation period and were fed a commercial lab chow diet after weaning. At 70 days, malnourished and control animals were placed on the central platform of the elevated plus-maze facing an enclosed arm and allowed to explore for 5 min. This procedure was repeated at 24-h intervals for 6 days. The repeated testing in the elevated plus-maze did not change the total number of arm entries and attempts to enter open arms, but decreased the percentage of open arm entries, time spent in open arms, and total time spent on the central platform. These data suggest an increase in anxiety with repeated testing in the elevated plus-maze. In addition, the malnourished animals showed a larger number of both rearings and attempts to enter the open arms, suggesting a high level of exploration and/or high impulsiveness of these animals as compared to control. The elevated plus-maze proved to be a useful animal model to evaluate exploratory behaviors in early protein malnourished animals.

Selection of an intact casein or casein hydrolysate diet by rats submitted to protein deprivation and bowel resection.

The feeding preference of normal rats (n = 14), malnourished rats (n = 14), and enterectomized rats (n = 16) was determined in a situation of free choice of three complete solid diets which only differed in extent of protein polymerization: intact casein, casein hydrolysate, and an amino acid mixture with a composition similar to that of casein. The animals were housed in metabolic cages for 30 days and allowed to freely choose among the three diets presented simultaneously. All three groups showed an initial preference for the intact casein diet. The control group maintained this preference, whereas the malnourished and enterectomized groups reduced the ingestion of the intact casein diet and increased the ingestion of the amino acid diet. The nitrogen balance, which was always positive in all three groups, was constant in the control group (1.51 +/- 0.26 g) initially higher (1.77 +/- 0.19 g) in the malnourished group, with a subsequent fall (1.13 +/- 0.24 g), and lower in the enterectomized group (0.83 +/- 0.32 g). Although total intake was similar for the control and malnourished groups, the malnourished group presented a higher weight recovery (130.2%). In contrast, the food intake of the enterectomized group was much lower, with a small weight gain. After treatment that impairs the digestive tract, intact casein was the initially preferred nitrogen source, which later tended to be replaced with free amino acids. This change was accompanied by an improvement in nitrogen balance and body weight, especially after protein malnutrition. These data may suggest that, in clinical practice, the use of enteral diets containing fully hydrolyzed protein may be of benefit in terms of the recovery of malnourished patients and of patients with short bowel syndrome.

Effects of early postnatal malnutrition and chlordiazepoxide on experimental aversive situations.

In order to study the lasting consequences of brain changes caused by early malnutrition, rats were fed a protein-deficient diet from birth until 49 days of age and a balanced diet from day 50 to day 70. At 49 and 70 days of age, independent groups of animals were tested in the locomotor activity, step-down inhibitory avoidance, and flinch-jump nociceptive tests. Also, at 49 days of age, malnourished and control rats were sacrificed in order to evaluate the weight of brain regions. Malnourished rats had lower body and brain weights (telencephalon and brain stem) than control rats. Malnourished rats also showed less locomotor activity at the beginning of the test session, lower flinch and jump thresholds, and longer step-down latencies than control animals. Chlordiazepoxide (5 mg/kg, IP) shortened step-down latency of well-nourished rats, but was ineffective in malnourished rats. These and previously reported results indicate that early protein malnutrition causes long-lasting impairment of neuronal systems underlying emotional behavior.

Effects of malnutrition during early lactation on development and feeding behavior under the self-selection paradigm.

Selection of food can be affected by several factors, and with the method of self-selection, qualitative changes in nutritional balance may be detected. The goal of the present study was to evaluate feeding preferences in weaning rats using three macronutrients (protein, carbohydrate, and fat), through a free-choice method, evaluating the alteration in their feeding patterns as compared with the previous nutritional status during the early lactation period. We analyzed the effects of protein restriction during lactation over the nitrogen balance after the weaning. The dams were assigned to one of two diet conditions (nourished or malnourished). At weaning, two pups from each litter were housed individually in metabolic cages, and they were maintained on self-selection under a free-choice paradigm and were provided with separate sources of macronutrients. The parameter for evaluating the nutritional effectiveness of the diets was nitrogen balance. We observed that protein intake tended to increase and consumption of carbohydrate and fat tended to decrease progressively during the 3 wk of experiment. In selecting their own food, growing rats and malnourished rats consumed a larger amount of protein than the other rats. Nourished rats selecting their diet had a larger nitrogen balance than nourished rats receiving a composed diet; no nitrogen balance difference was found between the self-selecting groups. Rats can choose an adaptive form when recovering from protein malnutrition.

Decreased reactivity to anxiolytics caused by early protein malnutrition in rats.

In order to investigate whether early malnutrition causes lasting changes in the reactivity to anxiolytic drugs, rat dams during lactation (21 days) and pups after weaning until the 49th day of life were fed on 8% casein diet (M rats), while their well-nourished controls received 25% casein (W rats). From day 50 on all animals ate the same balanced diet. Experiments started on the 91st day. Rats deprived for 22 hours drank water containing either 1.8% or 2.7% sodium chloride for 30 min in a test chamber, total intake being measured. Dose-effect curves for diazepam (0.5-5.0 mg/kg, IP), as well as for the nonbenzodiazepine anxiolytics ipsapirone (0.5-5.0 mg/kg), ritanserin (0.05-1.0 mg/kg) and isamoltane (2.5-20.0 mg/kg) were determined in M as well as in W rats. Diazepam and ipsapirone dose-dependently released drinking suppressed by either salt concentration in W rats, but caused little or no effect in M rats. Ritanserin and isamoltane were ineffective in both groups. These and previously reported results show that early protein malnutrition markedly reduces anticonflict effects of anxiolytics, indicating long-lasting impairment of neuronal systems underlying emotional behavior.

Effects of early protein malnutrition and environmental stimulation upon the reactivity to diazepam in two animal models of anxiety.

In order to investigate the effects of early protein malnutrition and environmental stimulation upon the response to the anxiolytic properties of diazepam, two animal models of anxiety (elevated plus-maze and light-dark transition tests) were used. Rats were malnourished by feeding their dams a 6% protein diet during the lactation period (0-21 days of age) while well-nourished controls received a 16% protein diet. From 21 to 70 days of age all rats received a balanced lab chow diet. Environmental stimulation consisted of 3-min daily handling from birth to 70 days of age. Additional stimulation was provided from 21 to 70 days of age by rearing the rats in an enriched living cage. Eight groups of rats were studied in a 2 (malnourished or well-nourished) x 2 (stimulated or nonstimulated) x 2 (diazepam or vehicle) design. At 70 days of age, independent groups of rats treated with diazepam (2.5 mg/kg, IP) or vehicle were submitted to testing in the elevated plus-maze or light-dark transition procedures. The results showed that both diazepam and environmental stimulation reduced anxiety in the elevated plus-maze; stimulation changed the anxiolytic response to diazepam and the two diet conditions altered differentially the response to both pharmacological and stimulation procedures. These results suggest that environmental stimulation can affect differentially the behavioral response of malnourished and well-nourished rats treated with diazepam.

Acquisition and extinction of jumping, two-way shuttle-box and bar press avoidance responses in malnourished rats: effects of shock intensity.

1. In order to investigate the role of avoidance response and shock intensity in avoidance learning in malnourished rats, three avoidance responses (jumping, two-way shuttle-box and bar press) and three shock intensities (0.4, 0.6 and 1.0 mA) were used. Independent groups of 6 rats were used for each response topography and shock intensity. 2. Malnourished male Wistar rats were suckled by mothers fed a 12% casein diet during the lactation period (0-21 days of age) while the mothers of well-nourished controls received a 25% casein diet. After weaning (21st day), all animals received a commercial lab chow diet until 70 days of age, when the avoidance training started. 3. Malnutrition did not affect the acquisition of the avoidance response, but malnourished groups required more trials to extinguish jumping and two-way shuttle-box. During the acquisition phase all animals learned the jump response faster in comparison to bar press and shuttle-box avoidance responses. Both groups in the acquisition phase responded faster with 1.0 mA when compared to lower intensities (0.6 and 0.4 mA). The malnourished animals showed lower latency of avoidance in the jumping response when compared with well-nourished animals. During the extinction phase there was a significant effect of diet, response topography and shock intensity in the latency to respond and trials to criterion. The increased resistance to extinction in malnourished rats was particularly evident with 1.0 mA in the two-way shuttle-box response. 4. These results suggest that contradictory data related to the acquisition of the avoidance response in malnourished animals cannot be attributed to response topography or variations in shock intensity. Furthermore, our results also indicate that resistance to extinction and latency to respond are appropriate parameters for detecting differences between well-nourished and malnourished animals.

Response threshold to aversive stimuli in stimulated early protein-malnourished rats.

Two animal models of pain were used to study the effects of short-term protein malnutrition and environmental stimulation on the response threshold to aversive stimuli. Eighty male Wistar rats were used. Half of the pups were submitted to malnutrition by feeding their mothers a 6% protein diet from 0 to 21 days of age while the mothers of the other half (controls) were well nourished, receiving 16% protein. From 22 to 70 days all rats were fed commercial lab chow. Half of the animals in the malnourished and control groups were maintained under stimulating conditions, including a 3-min daily handling from 0 to 70 days and an enriched living cage after weaning. The other half was reared in a standard living cage. At 70 days, independent groups of rats were exposed to the shock threshold or to the tail-flick test. The results showed lower body and brain weights in malnourished rats when compared with controls at weaning and testing. In the shock threshold test the malnourished animals were more sensitive to electric shock and environmental stimulation increased the shock threshold. No differences due to diet or environmental stimulation were found in the tail-flick procedure. These results demonstrate that protein malnutrition imposed only during the lactation period is efficient in inducing hyperreactivity to electric shock and that environmental stimulation attenuates the differences in shock threshold produced by protein malnutrition.

Effect of zinc deficiency induced before and during pregnancy on the survival of female rats and their pups.

The objective of the present study was to determine the consequences of Zn2+ deficiency on the gestational process. The study was conducted on adult Wistar virgin female rats fed isocaloric diets containing 16% protein and different Zn2+ concentrations, i.e., 2 ppm (severe restriction), 6 ppm (moderate restriction), and 20 ppm (control). Rats received the diets and water ad libitum for 7, 14 or 21 days before mating and throughout pregnancy. Survival of dams and pups decreased with increasing Zn2+ restriction and with time of exposure to the deficient diet. Mean survival rate for control dams and pups was 100%, whereas severe restriction (2 ppm for 21 days premating and during pregnancy) resulted in survival rates of 25% and 0 for dams and pups, respectively. Dam and pup survival rates for moderate restriction (6 ppm) for the same period were 83% and 72%, respectively. These results indicate the importance of Zn2+ before and during pregnancy and show that Zn2+ deficiency causes metabolic alterations which impair normal reproductive processes.

Food intake and weight of lactating rats maintained on different protein-calorie diets, and pup growth.

Studies on rats maintained on low-protein-calorie diets during the lactation period show that food intake decreases. This process results in weight loss and a delay in litter development. The purpose of the present study was to determine the alterations in food intake, maternal weight and litter growth during lactation when dams were exposed to diets with different levels of protein and carbohydrate. Female Wistar rats receiving one of 4 different diets, A (N = 14), B (N = 14), C (N = 9) and D (N = 9), were used. Diet A contained 16% protein and 66% carbohydrate; diet B, 6% protein and 77% carbohydrate; diet C, 6% protein and 66% carbohydrate; diet D, 16% protein and 56% carbohydrate. Thus, C and D diets were hypocaloric, while A and B were isocaloric. The intake of a low-protein diet in groups B and C affected the weight of dams and litters during the last two weeks of lactation, while the low-calorie diets limited the growth of D litters at 21 days compared with A litters, but had no effect on the weight of D dams. Group B showed an increase in intake during the first five days of lactation, resulting in a behavioral calorie compensation due to the increase in carbohydrate content, but the intake decreased during the last part of lactation. Food intake regulation predominantly involves the recruitment of a variety of peripheral satiety systems that attempt to decrease the central feeding command system.

Early malnutrition alters the effect of chlordiazepoxide on inhibitory avoidance.

In order to study the functional consequences of brain changes caused by early malnutrition, rats were fed a protein-deficient diet from birth until 49 days of age and a balanced diet from day 50 to day 70. The animals were submitted to a step-down inhibitory avoidance task and to the flinch-jump nociceptive test at 49 and 70 days of age. Malnourished rats showed longer step-down latencies and lower flinch and jump thresholds than eutrophic animals. Chlordiazepoxide (5 mg/kg, ip) shortened step-down latency of well-nourished rats, whereas it failed to do so in malnourished rats. Since well-nourished animals also became resistant to chlordiazepoxide when tested with a higher shock intensity, generating avoidance latencies comparable to those of malnourished animals, we conclude that the drug resistance induced by malnutrition may be secondary to enhanced pain sensitivity and/or reactivity.