940 resultados para plasma protein


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Currently, diagnostic tests for mesenteric ischaemia and infarction are inadequate due to poor sensitivity and specificity. In addition, many potential markers appear too late to be clinically useful. At present, definitive diagnosis can only be made at the time of surgery, which is not ideal as surgery is often to be avoided in critically ill and elderly patients. A clinically useful, minimally invasive test is likely to decrease the currently very high mortality rate and allow monitoring of 'at risk' patients during their hospital stay. A two-dimensional electrophoresis based proteomic approach was undertaken to assess plasma protein differences between patients with surgically confirmed bowel infarction and control Intensive Care patients. The major protein differences were found to be members or variants of acute phase proteins. Serum amyloid A showed the largest difference between the two patient groups, and this protein was investigated in greater depth. An analysis was performed to compare the diagnostic ability of several commonly used indicators of critical illness and bowel infarction with serum amyloid A and phospholipase A2. Although none of the variables were ideal for clinical use, plasma phospholipase A2 activity showed the best discriminatory power, as determined by Receiver Operating Characteristic curves. From a review of the literature, phospholipase AI (PLA2) appeared to be increased in the bowel as a result of ischaemia and infarction. In one patient, matched tissues were obtained, and PLA2 activity was found to be significantly higher in infarcted bowel tissue compared to ischaemic bowel tissue. PLA2 activity was significantly greater in bowel lumen than tissue, suggesting that the protein was being released, and may enter the circulation. PLA2 activity was increased in the plasma of bowel infarction patients compared with control patients, though the difference was not significant. The phospholipase activity exhibited a number of similarities to typical phospholipase A2 proteins, but also showed a number of inconsistent characteristics. For this reason, we wished to identify the protein responsible for the increased phospholipase activity in infarcted human bowel. The PLA2 activity in human bowel could not be abolished by immunoprecipitation of the PLA2 isoforms IIA (well described in bowel) and V (a closely related isoform). To investigate these proteins, a native urea protein gel devised for snake venom phospholipase A2 was modified for use with mammalian phospholipase AI. The modified gel was used to show that the protein with phospholipase activity from infarcted gut was different from normal gut PLA2 and type IIA PLA2. A number of extensions were devised for these native gels and were found to be useful both in this investigation and for venom investigations. Protein purification was undertaken to identify the protein responsible for the increased phospholipase activity in infarcted bowel. Protein was purified from infarcted human bowel using a number of techniques that exploited unusual characteristics of the protein. The purification techniques each retained the native activity of the protein and the purification could therefore be monitored with a phospholipid hydrolysis assay at each stage. The protein identified by mass spectrometry was an excellent match for cyclophilin B, an inflammatory protein that had previously been identified in rat bowel at the mRNA level (Hasel et al, 1991, Kainer & Doris, 2000). As the purification progress had been monitored throughout with a phospholipid hydrolysis assay, cyclophilin B was an unexpected identification, as it is not known to have phospholipase activity. Cyclophilin B was removed from the highly purified samples via immunoprecipitation and this process abolished all phospholipase activity. The addition of cyclosporin A, (the pharmaceutical ligand of cyclophilin B), did not effect the phospholipase activity. Cyclophilin B protein was found in normal and infarcted human bowel using Western blotting. Cyclophilin B protein also appeared to be present in the bowel lumen and plasma of several patients with bowel infarction, but not in control patients. Immunohistochemistry confirmed the ubiquitous nature of cyclophilin B that had been reported by other groups. This project has investigated the use of two dimensional gel electrophoresis based proteomics to identify proteins present in the plasma of patients with confirmed bowel infarction and control intensive care patients. The major protein classes observed were members of the acute phase proteins, which highlights the need for pre-fractionation of plasma to identify lower abundance, disease associated proteins. A series of potential plasma markers were compared using Receiver Operating Characteristic Curves. Although no ideal marker was clear from this analysis, phospholipase activity appeared to warrant further investigation. Phospholipase activity was investigated in human infarcted bowel. Protein purification identified cyclophilin B as a bowel protein that showed unusual phospholipid hydrolysing activity. Cyclophilin B is a ubiquitous protein in intestinal cell types in both normal and infarcted tissue. There appears to be release of cyclophilin B into bowel lumen and plasma under conditions of mesenteric ischaemia and infarction.

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Background: Hereditary angioedema (HAE) is a rare, debilitating, potentially life-threatening condition characterized by recurrent acute attacks of edema of the skin, face/upper airway, and gastrointestinal and urogenital tracts. During a laryngeal attack, people with HAE may be at risk of suffocation, while other attacks are often associated with intense pain, disfigurement, disability, and/or vomiting. The intensity of some symptoms is known only to the person experiencing them. Thus, interview studies are needed to explore such experience and patient-reported outcome measures (PROMs) are required for systematic assessment of symptoms in the clinical setting and in clinical trials of treatments for acute HAE attacks.

Objective: The aim of this interview study was to assess the content validity and suitability of four visual analog scale (VAS) instruments for use in clinical studies. The VAS instruments were designed to assess symptoms at abdominal, oro-facial-pharyngeal-laryngeal, peripheral, and urogenital attack locations. This is the first known study to report qualitative data about the patient's experience of the rare disorder, HAE.

Methods: Semi-structured exploratory and cognitive debriefing interviews were conducted with 27 adults with a confirmed clinical/laboratory diagnosis of HAE (baseline plasma level of functional plasma protein C1 esterase inhibitor [C1INH] <50% of normal without evidence for acquired angioedema). There were 17 participants from the US and 10 from Italy, with mean age 42.5 (SD 14.5) years, range 18–72 years, mean HAE duration 21.3 (SD 14.1) years, range 1–45 years, 67% female, and 44% VAS-naïve. Experience of acute angioedema attacks was first explored, noting spontaneous mentions by participants of HAE symptomatology. Cognitive debriefing of the VAS instruments was undertaken to assess the suitability, comprehensibility, and relevance of the VAS items. Asymptomatic participants completed the VAS instruments relevant to their angioedema experience, reporting as if they were experiencing an acute angioedema attack at the time. Interviews were conducted in the clinic setting in the US and Italy over an 8-month period.

Results: Participants mentioned spontaneously almost all aspects of acute angioedema attacks covered by the four VAS instruments, thus providing strong support for inclusion of nearly all VAS items, with no important symptoms missing. Predominant symptoms found to be associated with acute angioedema attacks were edema and pain, and there was evidence of varying degrees of disruption to everyday activities supporting the inclusion of an overall severity item reflecting the disabling effects of HAE symptoms. VAS item wording was understood by participants.

Conclusion: This interview study explored and reported the patient experience of HAE attacks. It demonstrated the content validity of the four anatomical location HAE VAS instruments and their suitability for use in clinical trials of recombinant human C1INH (rhC1INH) treatment for ascertaining trial participants' assessments of the severity of acute angioedema symptoms.

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Although many studies point to alterations in the organic concentrations of zinc in elderly patients, the mechanisms by which aging might cause changes in the metabolism of this nutrient remain unclear. Thus, we assessed the changes in plasma zinc, Zinc Binding Capacity to Plasma Protein (ZnBCPP) and Saturation Index (SI), comparing elderly individuals and young adults. The zinc analyses were performed by atomic absorption spectrophotometry. A statistically significant difference (p < 0.001) was found between the two groups, in relation to plasma zinc and SI, but the ZnBCPP did not differ between the younger and older subjects. In agreement with this result, it was shown in the young group that 76% (R2 = 0.760) of the ZnBCPP variations are explained by the variations in plasma zinc and SI. In the elderly group this measure decreased to 30.5% (R2 = 0.305). We conclude, therefore, that aging may be a factor associated to changes in control mechanisms and in zinc homeostasis, and could even alter ZnBCPP response patterns and other zinc-related indicators of nutritional status.

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A adição de sal à água tem sido utilizada para a mitigação de estresse e aumento da taxa de sobrevivência em peixes. O presente estudo avaliou o efeito do cloreto de sódio (0,0; 1,0; 3,0 e 6.0 g/l) nas concentrações de cortisol plasmático, glicemia, triglicerídios, proteínas total plasmática, hematócrito, hemoglobina, número de eritrócitos, glicogênio e lipídio hepáticos, e lipídio muscular em matrinxã Brycon amazonicum adultos após quatro horas de transporte e durante período de recuperação de 96 h. Amostras foram coletadas antes e depois do transporte, bem como 24 e 96 h após a chegada. O nível de cortisol plasmático estava mais elevado logo após o transporte quando comparado à condição inicial (pré-transporte), exceto para os peixes transportados com sal nas concentrações 3,0 e 6,0 g/l. Comportamento semelhante foi observado para a glicemia, porém os peixes dos tratamentos 0,0, 1,0 e 3,0 g/l necessitaram de período superior a 24 h para recuperar a condição inicial. Foram registrados níveis mais baixos de glicogênio hepático em peixes do tratamento controle (0,0 g/l). Os parâmetros hemoglobina, número de eritrócitos, proteína plasmática total e lipídio hepático não apresentaram alterações durante o período experimental. Os valores de hematócrito diminuíram logo após o transporte em todos os tratamentos, retornando aos níveis iniciais após 24 h. Todos os tratamentos apresentaram redução nos níveis de lipídio muscular e triglicerídios durante o período de recuperação. Os resultados sugerem que a adição de 6,0 g/l de sal na água de transporte reduz as alterações fisiológicas de estresse e que é necessário período de 96 h após o transporte para a recuperação da condição inicial de matrinxãs transportados sem a adição de sal.

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Foram examinados 20 eqüinos adultos, 10 sadios e 10 acometidos por abdômen agudo, submetidos à laparotomia. O exame clínico e a colheita de amostras de sangue foram realizados antes da laparotomia e diariamente, a partir da cirurgia, até o 10º dia após a intervenção. Constatou-se elevação da temperatura retal, das freqüências cardíaca e respiratória, do número de hemácias e de leucócitos, do volume globular e dos valores das proteínas plasmáticas após a cirurgia, em ambos os grupos, porém com valores mais elevados nos animais enfermos, especialmente do número de neutrófilos. O proteinograma plasmático dos eqüinos com abdômen agudo mostrou que houve elevação significativa nas concentrações de proteínas na fase aguda com maiores valores ao redor de 48 horas após a cirurgia. Os resultados indicaram que o padrão de elevação e decréscimo dessas proteínas pode ser útil na definição do prognóstico do quadro clínico de abdômen agudo e da recuperação cirúrgica dos eqüinos.

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Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

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The aim of this study was to evaluate and characterize phenotypically goats with different levels of resistance to gastrointestinal nematodes. For a period of 93 days, 60 F2 goats originated from ½ Saanen and ½ Anglo- nubian animals were kept in the same area of pasture. Every seven days, feces and blood were collected for eggs per gram counts of feces (EPG) and cultures of feces and to determinate the number of eosinophils, packed cell volume and total plasma protein, respectively. On the same day, the animals were weighed and submitted to body score condition and FAMACHA method to worm control. Based on the average of EPG, the twelve animals with the highest average (susceptible group) and the twelve animals with the lowest average of EPG (resistant group) were selected, slaughtered and necropsied to recovery, counting andparasites identification. The resistant animals present lower EPG mean (P <0.0001) and 4.7 folder less parasites than susceptible animals. The resistant group presented higher mean packed cell volume (26.48%) and total plasma protein (6.24 g / dl) than susceptible one (24,04% e 5,82g/dl, respectively). The average number of eosinophils was similar in both groups The Haemonchus sp. was the most prevalent in the culture of feces, followed by Trischostrongylus sp. and Oesophagostomum sp.. The counting of nematodes in the abomasum of susceptible group was higher than in resistant one. The species identified were H. contortus in abomasums and T. colubriformis in small intestine. It can be concluded that EPG, packed cell volume and total plasma protein were useful phenotypic markers to identify animals as resistant and susceptible to gastrointestinal nematodes infections

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O objetivo do presente estudo foi realizar a comparação entre dois sítios de coleta sanguínea em 24 exemplares de tigre-d'água-americano (Trachemys scripta elegans) oriundos de um criadouro comercial, localizado no município de Antonina, litoral do Paraná, Brasil. Os animais foram submetidos a contenção física e as venopunções foram realizadas no seio supraocciptal e na veia coccígea dorsal. As amostras heparinizadas foram identificadas e refrigeradas para posterior análise laboratorial. A contagem total de eritrócitos e leucócitos foi realizada pela técnica de hemocitometria. O hematócrito (Ht) e a hemoglobina (Hb) foram determinados pelo método de microhematócrito e cianometahemoglobina, respectivamente. A proteína plasmática total (PPT) foi determinada por refratometria e a contagem diferencial de leucócitos foi realizada através da técnica de Shilling. Houve diferença significativa no número de leucócitos e no valor da proteína plasmática total, e em ambos os casos os valores encontrados nas amostras provenientes da veia coccígea dorsal foram inferiores. A diferença encontrada no número de leucócitos provavelmente foi devido à contaminação por linfa, que também justifica o menor valor na concentração da proteína plasmática total. Conclui-se que é mais indicada a venopunção no seio supraocciptal quando comparado a veia coccígea dorsal.

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O presente trabalho teve como objetivos avaliar os sinais clínicos, as concentrações do sulfeto de hidrogênio ruminal e as alterações anatomopatológicas associadas à intoxicação experimental por enxofre em bovinos. Foram utilizados dez bezerros mestiços leiteiros, sendo que quatro bovinos ingeriram ração sem sulfato de sódio (G1) e seis consumiram ração com sulfato de sódio (G2). Exames clínicos (temperatura retal, frequência cardíaca e respiratória e motricidade ruminal) e laboratoriais (hemograma, fibrinogênio, proteína plasmática, pH do fluido ruminal, concentração do sulfeto de hidrogênio ruminal, líquido cerebrospinal e histopatológico) foram realizados. A temperatura retal, frequência cardíaca, hemograma, fibrinogênio, proteína plasmática, pH do fluido ruminal e os valores do líquido cerebrospinal estavam dentro dos valores de referência para a espécie. Taquipnéia, hipomotricidade ruminal e elevados valores de sulfeto de hidrogênio ruminal foram observados nos bezerros do grupo G2. Um bezerro do grupo G2 apresentou sinais neurológicos e lesões histopatológicas de PEM. Dois animais de cada grupo foram eutanasiados. Lesões microscópicas foram observadas nos bezerros do G2. Histologicamente as alterações observadas foram necrose neuronal cortical e lesões hemorrágicas nos núcleos basais, tálamo, mesencéfalo, ponte e bulbo. O protocolo experimental constituído por uma dieta rica em carboidrato de alta fermentação, baixa quantidade de fibra efetiva e altos níveis de enxofre (0,52%) ocasionou alterações clinicas e histológicas e elevadas concentrações de sulfeto de hidrogênio ruminal compatíveis com quadro de intoxicação por enxofre.

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Natural (NV) and Cobalto60-irradiated (IrV) Crotalus durissus terrificus venom were used to evaluate serum production capacity of sheep and possible hematological and biochemical effects. Freeze-dried venom aliquots were diluted in acidified saline solution (NaCl 150 mM, pH 3.0) and irradiated by a Cobalt 60 source at a dose of 5.54 x 102 Gy/h and a concentration of 2.000 Gy. Twelve sheep were divided into two groups of six animals. One group received irradiated venom (IrV) and the other natural venom (NV). Three antigen doses (venom) were administered at monthly intervals. Blood samples were collected weekly for analysis of serum neutralization potency and capacity, complete blood count (CBC), total plasma protein, fibrinogen, albumin, and globulin. At the end of the experiment, the animals were challenged with a LD50 for sheep and showed no signs of envenoming. The two groups did not present clinical alterations. Results of the total leukocyte count did not present interaction or time factor effect for both groups, but there was a different action between them, with the NV group presenting more cells than the IrV group. The leukocyte increase to 13,000/ml indicates that slight leukocytosis occurred in the week after the first inoculation in the NV group. There was no statistically significant difference between groups in the absolute count of segmented neutrophils, eosinophils, and lymphocytes but there were statistically significant oscillations in values at the different collecting times. The NV group presented an increase in the absolute neutrophil count after the first inoculation that persisted for 5 weeks. In the IrV group, the increase in neutrophils occurred only in the first week returning to normal in the following weeks. The alterations in the neutrophil count are indicative of systemic inflammatory response related to cytokine release; response was more marked in the NV group, showing its greater toxicity.

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O grau de contaminação da pastagem por larvas de terceiro estágio (L3) de nematoides gastrintestinais foi avaliado em piquetes pastejados por ovelhas resistentes ou susceptíveis à verminose. O experimento foi realizado de 23 de novembro de 2007 a 22 de julho de 2008. Inicialmente, realizou-se a contagem de ovos por grama de fezes (OPG) de amostras individuais de 130 ovelhas adultas da raça Bergamácia. Dessas, foram selecionadas as nove ovelhas com contagens mais elevadas de OPG (susceptíveis) e as 10 com as menores contagens (resistentes). As ovelhas resistentes apresentaram menor contagem de OPG, maiores valores de volume globular, de proteína plasmática total e de eosinófilos sanguíneos, do que as ovelhas susceptíveis. O peso também foi maior no grupo resistente. Na pastagem, foram identificadas larvas de Haemonchus spp., Trichostrongylus spp. e Oesophagostomum spp. em média, as quantidades de L3 de Haemonchus spp. e de Trichostrongylus spp., na pastagem, foram 2,19 e 2,31 vezes, respectivamente, maiores nos piquetes pastejados pelo grupo susceptível do que nos do grupo resistente. Portanto, os animais susceptíveis devem ser eliminados do rebanho a fim de reduzir a contaminação da pastagem e otimizar a profilaxia das infecções por nematoides gastrintestinais.

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The response to natural infection by gastrointestinal nematodes was evaluated in three groups of lambing mature ewes: 16 Florida Native; 21 Rambouillet; and 18 1/2 Florida Native x 1/2 Rambouillet (F1 generation). Fecal egg counts (FEC),packed cell volume (PCV) and plasma protein levels were determined periodically through a one-year period. The highest FEC was recorded in Rambouillet ewes, with most of the means higher than 1000 eggs per gram (EPG). Florida Native and Fl ewes showed similar trends in FEC, with most of the means below 500 EPG. The highest mean PCV values were recorded in Florida Native ewes, followed in decreasing order by Fl and Rambouillet ewes. Fl ewes showed the lowest plasma protein levels and Florida Natives the highest. Fecal cultures revealed the highest percentages of Haemonchus larvae in Rambouillet ewes (>64%) and the lowest percentages in Florida Native ewes. Florida Native and Fl ewes were more resistant to gastrointestinal nematode infections than were Rambouillet ewes. High negative correlation coefficients between FEC x PCV and FEC x plasma protein levels coincided with peaks in mean FEC of Rambouillet (May and July), Florida Native and Fl ewes (October). This study showed that crossbreeding Rambouillet and Florida Native sheep resulted in the production of Fl ewes resistant against gastrointestinal nematode infections. (C) 1999 Elsevier B.V. B.V. All rights reserved.

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Breeds of sheep with resistance to nematode infection often display a reduced periparturient rise (PPR) in faecal egg counts (FEC) when compared with susceptible sheep. This trial was carried out to compare the resistance of Santa Ines, an indigenous Brazilian breed of sheep, and Ile de France ewes to natural infections by gastrointestinal nematodes, especially in the periparturient period. Twelve Santa lnes and 10 Ile de France ewes were estrus synchronized and then mated (December 2001). Lambing occurred from 30 April to 19 June 2002 and the lambs were weaned at 60 days of age. Faecal egg counts, packed cell volume (PCV), total plasma protein levels and peripheral eosinophils counts were determined from November 2001 to September 2002. To prevent deaths, individual treatment with anthelmintics was provided to ewes with FEC higher than 4000 eggs per gram (EPG) or with PCV lower than 21%. The percentage of Santa Ines ewes treated with anthelmintics was lower than that of Ile de France sheep. Eight of the 10 Ile de France ewes were treated, with one of them requiring four treatments, one requiring three treatments, and two requiring two treatments. The remaining four received a single treatment. In contrast, only 5 of the 12 Santa lnes ewes required treatment and for each this was done only once. The mean values of PCV were in the normal range in the Santa Ines ewes. In contrast, mean PCVs of Ile de France ewes reached a low of 24.5% in the second month after lambing and were significantly lower than Santa Ines 1 month before lambing (P < 0.05) and 2 months after lambing (P < 0.07). The mean plasma protein level was significantly lower in the Ile de France ewes in the second month after lambing (P < 0.01) coinciding with a reduction in PCV. The ewes of both breeds had high mean numbers of eosinophils in blood, which increased even further post-weaning when a marked reduction in FEC occurred. Haemonchus larvae were the most numerous genus in faecal cultures for both breeds followed by Trichostrongylus, Oesophagostomum and Cooperia. A periparturient rise in FEC was observed in both breeds. However, Santa Ines ewes showed a higher capacity to support the infection in an environment contaminated with large number of infective larvae of gastrointestinal nematodes. (C) 2004 Elsevier B.V. All rights reserved.

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This experiment was carried out to compare the worm burden and cellular responses in the abomasal mucosa and blood of Florida Native and Rambouillet lambs and also in animals produced by crosses of these two breeds (generations F1 and F2). Animals were exposed to infection by gastrointestinal nematodes on three different occasions. The first infection was natural, occurring while they were suckling lambs. After weaning, they were kept indoors for 53 days and then were allowed to graze a contaminated pasture for 50 days for a second natural infection. The third infection was an artificial challenge with 6000 Haemonchus contortus infective larvae. The highest mean fecal egg counts (FEC) values were found in Rambouillet lambs followed in decreasing order by F1, F2 and Florida Native lambs. Throughout the trial, most of the high mean packed cell volumes and plasma protein levels were recorded in the F2 lambs; in contrast, most of the low values were found in the Rambouillet lambs. During the natural infection period, the highest percentages of larvae in the fecal cultures of the lambs were Haemonchus. However, high percentages of Trichostrongylus larvae were found particularly in Florida Native lambs. The mean number of blood eosinophils increased after the artificial challenge, reached a peak 21 days after infection and then declined. The highest and lowest blood eosinophil means were recorded in F2 and Florida Native lambs, respectively. The H. contortus burden was significantly higher in Rambouillet and in F1 lambs than in Florida Native and F2 lambs (p < 0.05), while no significant differences were recorded among eosinophil, mast cell and globule leucocyte counts in the abomasal mucosa (p > 0.05). The highest correlation coefficient recorded at the end of this study was between FEC and worm burden (r = 0.7). These two parameters showed a moderate negative correlation with PCV, plasma protein and mast cell counts in the abomasal mucosa. The results obtained in this study indicate that crossbreeding Florida Native and Rambouillet sheep can be a rapid way to combine and improve the characteristics of these two breeds. The parasitological results were promising. however, more studies will be necessary to verify the impact of crossbreeding in other traits. (C) 1999 Elsevier B.V. B.V. All rights reserved.

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A study was conducted to assess the breed resistance against nematode infections in Santa Ines, Ile de France and Suffolk male lambs over a 9-month period in São Paulo state, Brazil. Lambs were born during the winter (year 2000) and were weaned at 2 months of age. The animals were then housed and treated with anthelmintics to eliminate natural infections by gastrointestinal nematodes. In late October 2000, lambs were placed in a paddock, where they stayed until August of the following year. Fecal and blood samples were taken from each animal every 2 weeks. on the same day, a pasture sample was collected to determine the number of infective larvae on the herbage. To prevent deaths, individual treatment with anthelmintics was provided to lambs with fecal egg counts (FEC) higher than 4000 eggs per gram (EPG) or with a packed cell volume (PCV) lower than 21%. In August 2001, all animals were slaughtered and the worms present in samples of the gastrointestinal contents were identified and counted. Most of the Suffolk and Ile de France sheep received three to six anthelmintic treatments over a period of 7 months, while most of the Santa Ines were not treated. Reductions in PCV and plasma protein values associated with high FEC and worm burdens were recorded, particularly, in Suffolk and Ile de France lambs. Haemonchus contortus and Oesophagostomum columbianum burdens and number of nodular lesions caused in the large intestine by O. columbianum larvae were significantly lower in Santa Ines sheep. All three breeds showed similar Trichostrongylus colubriformis worm burdens. The relative resistance of Santa Ines young male sheep was superior to that of Suffolk and Ile de France sheep. (C) 2004 Elsevier B.V. All rights reserved.