Hematological changes in sheep inoculated with natural and Cobalt60-irradiated Crotalus durissus terrificus venom (Laurenti, 1768)

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.

Ocorrência de lesões hepáticas provocadas por larvas de Strongylus spp diagnosticadas no exame post mortem de eqüídeos abatidos em Apucarana, Paraná

The objective of this study was to determinate the occurrence of hepatic lesions caused by migratory large Strongyle larvaes in horses slaughtered in city of Apucarana in state of Parana. The lesions were diagnostic in post mortem exam by macro and microscopic analysis. From April 2003 to April 2004,38,363 animals, coming from different regions of Brazil, were examined. The occurrence of granulomas in liver was observed in 14,443 (37.64%), with adhesions and spot in 6,645 (17,32%), and 17,275 (45.03%) without macroscopic lesions. Macroscopic analysis revealed the presence of livers with calcified nodules, the presence of whitish spots, and adhesions in the format of "lines" over the hepatic capsule. No larvae were found in the livers. Hepatic fragments were histologically processed and revealed, by optical microscopy, inflammatory cells with predominance of eosinophils around the granulomas with a moderate amount of macrophages and the presence of fibroblasts.

Contaminação da pastagem com larvas infectantes de nematoides gastrintestinais após o pastejo de ovelhas resistentes ou susceptíveis à verminose

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.

Leukotriene B(4) is essential for selective eosinophil recruitment following allergen challenge of CD4(+) cells in a model of chronic eosinophilic inflammation

Subcutaneous heat-coagulated egg white implants (EWI) induce chronic, intense local eosinophilia in mice, followed by asthma-like responses to airway ovalbumin challenge. Our goal was to define the mechanisms of selective eosinophil accumulation in the EWI model. EWI carriers were challenged i.p. with ovalbumin and the contributions of cellular immunity and inflammatory mediators to the resulting leukocyte accumulation were defined through cell transfer and pharmacological inhibition protocols. Eosinophil recruitment required Major Histocompatibility Complex Class It expression, and was abolished by the leukotriene B4 (LTB4) receptor antagonist CP 105.696, the 5-lipoxygenase inhibitor BWA4C and the 5-lipoxygenase activating protein inhibitor MK886. Eosinophil recruitment in EWI carriers followed transfer of: a) CD4(+) (but not CD4(-)) cells, harvested from EWI donors and restimulated ex vivo; b) their cell-free supernatants, containing LTB4. Restimulation in the presence of MK886 was ineffective. CC chemokine receptor ligand (CCL)5 and CCL2 were induced by ovalbumin challenge in vivo. mRNA for CCL17 and CCL11 was induced in ovalbumin-restimulated CD4(+) cells ex vivo. MK886 blocked induction of CCL17 Pretreatment of EWI carriers with MK886 eliminated the effectiveness of exogenously administered CCL11, CCL2 and CCL5. In conclusion, chemokine-producing, ovalburnin-restimulated CD4(+) cells initiate eosinophil recruitment which is strictly dependent on LTB4 production. (C) 2008 Elsevier B.V. All rights reserved.

The role of chemokines and chemokine receptors in eosinophil activation during inflammatory allergic reactions

Chemokines are important chemotactic cytokines that play a fundamental role in the trafficking of leukocytes to sites of inflammation. They are also potent cell-activating factors, inducing cytokine and histamine release and free radical production, a fact that makes them particularly important in the pathogenesis of allergic inflammation. The action of chemokines is regulated at the level of agonist production and processing as well as at the level of receptor expression and coupling. Therefore, an analysis of the ligands must necessarily consider receptors. Eosinophils are target cells involved in the allergic inflammatory response since they are able to release a wide variety of mediators including CC and CXC chemokines and express their receptors. These mediators could damage the airway epithelial cells and might be important to stimulate other cells inducing an amplification of the allergic response. This review focuses on recently emerging data pertaining to the importance of chemokines and chemokine receptors in promoting eosinophil activation and migration during the allergic inflammatory process. The analysis of the function of eosinophils and their chemokine receptors during allergic inflammation might be a good approach to understanding the determinants of asthma severity and to developing novel therapies.

Biocompatibility Evaluation of 3 Facial Silicone Elastomers

The failure of facial prostheses is caused by limitations in the properties of existing materials, especially the biocompatibility. This study aimed to evaluate the biocompatibility of maxillofacial silicones in subcutaneous tissue of rats. Thirty Wistar rats received subcutaneous implants of 3 maxillofacial silicone elastomers (LIM 6050, MDX 4-4210, and industrial Silastic 732 RTV). A histomorphometric evaluation was conducted to analyze the biocompatibility of the implants. Eight areas of 60.11 mm(2) from the surgical pieces were analyzed. Mesenchymal cells, eosinophils, and foreign-body giant cells were counted. Data were submitted to analysis of variance and Tukey test. Initially, all implanted materials exhibited an acceptable tissue inflammatory response, with tissue reactions varying from light to moderate. Afterward, a fibrous capsule around the silicone was observed. The silicones used in the current study presented biocompatibility and can be used for implantation in both medical and dental areas. Their prosthetic indication is conditioned to their physical properties. Solid silicone is easier to adapt and does not suffer apparent modifications inside the tissues.

Relationship of abomasal histology and parasite-specific immunoglobulin A with the resistance to Haemonchus contortus infection in three breeds of sheep

This study was carried out to evaluate the relationship of abomasal inflammatory cells and parasite-specific immunoglobulin A (IgA) in mucus, with the resistance to Haemonchus contortus infection in three breeds of sheep naturally infected with gastrointestinal nematodes. The breeds were the native Santa Ines sheep, and the European Suffolk and Ile de France breeds. Mast cells, eosinophils and globule leucocytes were enumerated in abomasal mucosa. Eosinophils within the sub-mucosa also were counted separately. Histamine concentration was estimated in abomasal tissue samples. Enzyme-linked immunosorbent assay was carried out in mucus samples to determine the level of IgA anti-H. contortus third and fifth instar. There were no significant differences among group means of these variables (P > 0.05). The correlation coefficients between fecal egg counts (FEC) x mast cells (r = -0.490; P < 0.05) and FEC x eosinophils in sub-mucosa (r = -0.714; P < 0.01) was significant in the Santa Ines sheep. In the Ile de France group, the correlation coefficients between globule leucocytes x FEC (r = -0.879; P < 0.001) and histamine x worm burden (r = -0.833; P < 0.01) were also significant. In the Santa Ines and Ile de France sheep, correlation coefficients between IgA anti-L3 x worm burden and IgA anti-L3 x FEC were negative. In general, inflammatory cells and IgA-parasite-specific in abomasum were inversely associated with H. contortus worm burden and FEC indicating that they may impair parasite development or fecundity in the three breeds of sheep. However, similar mean values of inflammatory cells and IgA were found in the resistant (Santa Ines) and in the susceptible (Suffolk and Ile de France) breeds of sheep. The enumeration of cells by histological assessment does not provide information on their functional activity, which may be different among breeds. Thus, the effect of breed on the functional activity of these and other inflammatory cells is an important area for further study. (c) 2004 Elsevier B.V. All rights reserved.

Cytochemical characterization of eosinophilic leukocytes circulating in the blood of the turtle (Chrysemys dorbignih)

Eosinophils and neutrophils are granulocytic leukocytes that are present in the blood of most vertebrates. Studies have been performed on lower vertebrates to understand the biological roles of the cells in defense mechanisms and to establish phylogenetic studies and new experimental models. Whether these 2 cell types exist in reptiles is a matter of controversy. In the blood of turtles there are 2 types of granulocytes that exhibit eosinophilia, one of them with round cytoplasmic granules and the other with elongated cytoplasmic granules. It has been suggested that these cells may be eosinophils in different stages of maturation but they also may be distinct cell types, i.e. eosinophils and neutrophils. In the present study, we characterized the 2 types of granulocytes that are present in the blood of Chrysemys dorbignih, using cytochemical techniques. Type I eosinophils showed activity of nonspecific esterase, peroxidase activity that is resistant to KCN, and basic proteins. Type II eosinophils exhibited activity of trimetaphosphatase, alkaline phosphatase, nonspecific esterase, peroxidase that is sensitive to KCN, and basic proteins. These observations indicate the existence of 2 distinct cell types in the blood of Chrysemys dorbignih, type I and type II eosinophils, that correspond to eosinophils and heterophils (neutrophils) of mammals and other vertebrates.

Inflammatory reaction to the human bot-fly, Dermatobia hominis, in infested and reinfested mice

Two groups of mice were infested with first stage larvae of the human bot-fly, Dermatobia hominis (Linnaeus Jr) (Diptera: Oestridae). In the first group, skin biopsies were carried out 1, 3, 5, 7, 10 and 18 days after infestation. The second group was also infested but had all the larvae removed 5 days after infestation. The mice in the latter group were reinfested 4 weeks later and skin biopsies were carried out 1, 3, 5, 7, 10 and 18 days after reinfestation. In the first group, an inflammatory reaction began slowly, the neutrophils being the main inflammatory cells, eosinophils being scarce. The reaction progressed with time, developing a necrotic halo around the larvae containing inflammatory cells surrounded by fibroblasts. The inflammation invaded the adjacent tissue. In the second group, the inflammatory reaction was intense on the day immediately after reinfestation, the pattern being changed by the presence of a large number of eosinophils. Activated fibroblasts surrounding the necrotic area around the larvae appeared 3 days after reinfestation in the second group and 7 days after infestation in the first group. The results demonstrated that the previous contact with the antigens elicited the early arrival of eosinophils, probably through the chemotactic factors liberated by mast cells in the anaphylactic reaction.

Comparison of the susceptibility of Santa Ines and Ile de France ewes to nematode parasitism around parturition and during lactation

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.

Nematode burdens and cellular responses in the abomasal mucosa and blood of Florida Native, Rambouillet and crossbreed lambs

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.

Relationship of intestinal histology with the resistance to Trichostrongylus colubriformis infection in three breeds of sheep

Avaliaram-se a associação entre o número de células inflamatórias no intestino delgado e a resistência à infecção por Trichostrongylus colubriformis em ovinos de três raças (Santa Inês, Suffolk e Ile de France), naturalmente infectados. Mastócitos, eosinófilos e leucócitos globulares foram quantificados na mucosa intestinal. A concentração de histamina foi estimada em amostras teciduais do intestino, bem como foi determinado o comprimento de machos e fêmeas de T. colubriformis. A resposta celular foi similar na mucosa intestinal das três raças ovinas (P>0,05). Houve grande variação entre os ovinos em relação aos resultados parasitológicos e celulares, mesmo nos animais de mesma raça. em geral, os animais que apresentaram número menor de células inflamatórias tiveram cargas parasitárias maiores, contagens de ovos por grama de fezes mais altas e exemplares de T. colubriformis maiores. Os resultados indicaram que mastócitos, eosinófilos e leucócitos globulares prejudicaram o estabelecimento, o desenvolvimento e a sobrevivência dos parasitas.

Parasitological characteristics and tissue response in the abomasum of sheep infected with Haemonchus spp.

Realizou-se um estudo histopatológico da mucosa do abomaso de 40 cordeiros da raça Corriedale, expostos à infecção natural por Haemonchus spp. Os cordeiros foram colocados em pastagens contaminadas por 14 dias e após 28 dias de estabulação foram necropsiados. Por ocasião da necropsia, foram colhidas amostras de fezes para a realização da contagem de ovos por grama de fezes (OPG) e amostras do abomaso para exame histopatológico e contagem do número de eosinófilos, mastócitos e leucócitos globulares. O número de Haemonchus spp. presente no abomaso foi estimado a partir de uma alíquota de 10% do conteúdo. O número de ovos por fêmea foi estimado em 10 fêmeas de Haemonchus spp. colhidas de cada animal. Coeficientes de correlação significativos entre as características analisadas foram: número de Haemonchus e contagem de OPG (r = 0,86); comprimento de fêmeas e número de ovos por fêmea (r = 0,60); comprimento de fêmeas e contagem de OPG (r = 0,53); número de eosinófilos e número de Haemonchus (r = 0,48); número de eosinófilos e número de leucócitos globulares (r = 0,54); número de mastócitos e comprimento de fêmeas (r = -0,39) e número de mastócitos e número de leucócitos globulares (r = 0,34). Das características analisadas a que apresentou maior correlação com a carga parasitária dos animais foi a contagem de OPG.

Cytokine gene expression in response to Haemonchus placei infections in Nelore cattle

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

Parasitism by Oestrus ovis: Influence of sheep breed and nematode infections

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