Quantitative proteomic analysis in symptomatic and asymptomatic apical periodontitis.

AIM: To quantitatively and qualitatively compare the host proteomic profile in samples of symptomatic and asymptomatic apical periodontitis (AP) using nano-liquid chromatography-electron spray tandem mass spectrometry. METHODOLOGY: Samples were obtained from 18 patients with radiographically evident AP, divided into symptomatic and asymptomatic groups (nine per group) according to clinical characteristics. After sample collection, protein extraction, purification and quantification of the samples were performed, which were analysed by reverse-phase liquid chromatography coupled to mass spectrometry. Label-free quantitative proteomic analysis was performed by Protein Lynx Global Service software. Differences in expression of proteins between the groups were calculated using the Monte Carlo algorithm, considering P < 0.05 for down-regulated proteins and 1 - P > 0.95 for up-regulated proteins. Proteins were identified with the embedded ion accounting algorithm in the software and a search of the Homo sapiens UniProt database. RESULTS: A total of 853 individual human proteins were identified. In the quantitative analysis, common proteins to both groups accounted for 143 proteins. Differences in expression between groups resulted in 51 up-regulated proteins (1 - P > 0.95) in the symptomatic group, including alpha-1-antitrypsin, protein S100-A8, myeloperoxidase, peroxiredoxin and lactotransferrin. This group also had 43 down-regulated proteins (P < 0.05), comprising immunoglobulin, neutrophil defensin, pyruvate kinase and alpha-enolase. The qualitative analysis considered only the exclusive proteins of each group. For the symptomatic group, 318 complete proteins and 29 fragments were identified, such as dedicator of cytokinesis protein, intersectin, prostaglandin, phospholipase DDHD2 and superoxide dismutase. For the asymptomatic group, 326 complete proteins and 37 fragments were identified, including azurocidin, C-reactive protein, collagen alpha, cathepsin, heat shock and laminin. CONCLUSIONS: Quantitative differences in the expression of common proteins in cases of symptomatic and asymptomatic AP were found, which were mostly related to host immune response in both groups. Exclusive proteins in the symptomatic group were mainly related to the host response to the presence of viruses in endodontic infections, oxidative stress and proteolytic enzymes. The results provide a basis for a better understanding of cellular and molecular pathways involved in AP, establishing specific proteomic profiles for symptomatic and asymptomatic conditions.

Haematological response of curimbas Prochilodus lineatus, naturally infected with Neoechinorhynchus curemai.

This study evaluated the haematological response of curimbas Prochilodus lineatus, naturally infected with Neoechinorhynchus curemai (Acanthocephala: Neoechinorhynchidae). Thirty-seven fish were captured in October 2010 from the Mogi Guaçu River, Porto Ferreira, SP, Brazil. Infected fish presented increased mean corpuscular volume of erythrocytes, and lower thrombocyte and higher monocyte counts than uninfected fish.

Transplacental transmission of Toxoplasma gondii in reinfected pregnant female canines.

Twelve pregnant female canines, naturally infected with Toxoplasma gondii, were reinfected with T. gondii: three (GI) received tachyzoites subcutaneously (1.0 x 107), three (GII) were orally inoculated with oocysts (1.5 x 104), and six (GIII) were kept as a nonreinfected control group. All the reinfected female canines (GI and GII) miscarried or presented fetal death, while only one GIII female presented a stillborn in a litter of four pups (P < 0.01). Fever, lymphoadenopathy, miscarriage, and fetal death were the main clinical alterations observed. The highest serological titers detected through the indirect fluorescence antibody test (IFAT) were 1,024 (GI) and 4,096 (GII). In group III, the titers ranged between 64 and 256. By bioassays in mice, T. gondii was isolated in 17 organs of the reinfected adult canines, in 11 of the control group, and in 20 of the neonates. Positive immunostaining of cysts and/or tachyzoites were observed in 26 canine tissues (14 from GI and GII and ten from GIII). The agent was detected by immunohistochemistry in the encephalon of a neonate and in the spinal cord of a stillborn, thus, confirming that T. gondii infected canine fetuses, provoking miscarriages, even in bitches that presented primoinfection.

Effects of glycosaminoglycan polysulphate on the organisation of collagen fibres in experimentally induced tendonitis in horses.

An inflammatory process was induced by intratendinous injection of bacterial collagenase into the superficial digital flexor tendon (SDFT) of the left thoracic limb of 10 horses. One week later, the tendons in five of the horses (group 1) were treated with glycosaminoglycan polysulphate (GAGPS), and the tendons of the other five (group 2) were treated with saline solution. The horses were euthanased 150 days after the collagenase injections, and samples of the SDFTs were frozen at -14 degrees C, sectioned at 5 to 7 mum longitudinally and transversely, and stained by the picrosirius red method. Morphometric analysis was used to quantify the organised and disorganised bundles of collagen in the samples from groups 1 and 2. Significantly more organised bundles of collagen were observed in the tendons treated with GAGPS.

Fluoride concentration and amount of dentifrice influence enamel demineralization in situ.

OBJECTIVES: This study evaluated the effect of conventional (CD, 1100ppm F) and low-fluoride (LFD, 550ppm F) dentifrices, applied in different quantities, on enamel demineralization, and on fluoride (F) concentrations in the dental biofilm formed in situ. METHODS: Five combinations of dentifrices and quantities were tested: placebo (P-F-free) applied on all brush bristles; LFD applied by the transversal technique (0.3g-T1) or on all bristles (0.6g-T2); and CD applied in a pea-sized amount (0.15g-T3) or by the transversal technique (0.3g-T4), in order to produce comparable intensities (F concentration in the dentifrice×amount applied to the brush). Volunteers (n=13, 20-36 years old) wore palatal devices containing 4 bovine enamel blocks, and performed cariogenic challenges (30% sucrose solution) 6×/day, and brushing 3×/day, following a double-blind, cross-over and randomized protocol. On the 8th day, biofilm was collected 5 and 60min after brushing. The percentage of surface hardness loss (%SH), integrated loss of subsurface hardness (ΔKHN) and biofilm F concentrations (solid and fluid phases) were determined. Data were analyzed by repeated-measures ANOVA, Student-Newman-Keuls test, and Pearson's correlation coefficient (p<0.05). RESULTS: Significantly lower ΔKHN was observed for treatments with higher intensity (T2 and T4) in comparison with the lower intensity (T1 and T3). A strong correlation was observed between ΔKHN and F concentrations in total biofilm (r=-0.71) and biofilm fluid (r=-0.72) 5min after brushing. CONCLUSIONS: The treatment intensity has a significant influence on the development of caries lesions in situ. CLINICAL SIGNIFICANCE: The intensity of treatment (amount of dentifrice×concentration) during brushing seems to be a more relevant parameter of clinical efficacy than simply observing the F concentration of the product. The use of a small amount of CD significantly reduced the protective effects against enamel demineralization.

Morphological, cytochemical, and ultrastructural study of thrombocytes and leukocytes in neotropical fish, Brycon orbignyanus Valenciennes, 1850 (Characidae, Bryconinae).

Morphological, cytochemical and ultrastructural studies are important to demonstrate the function of the blood cells, which is very little understood in teleosts. In peripheral blood of 'piracanjuba' Brycon orbignyanus, thrombocytes, lymphocytes, monocytes, neutrophils and heterophils were studied and characterized. Thrombocytes had a fusiform or oval shape with PAS-positive granules. Lymphocytes presented small size with sparse basophilic cytoplasm. Monocytes were large in size, presented basophilic cytoplasm that may be foamy or vacuolated, with non-specific esterase staining. The neutrophils presented lightly neutrophilic granule cytoplasm, with positivity for PAS and peroxidase. The heterophils were large in size, with eosinophilic and basophilic granules cytoplasm and PAS-positive. Transmission electron microscopy study demonstrated that the thrombocytes, lymphocytes and monocytes features were similar to other teleosts. In ultrastructural study only one type of neutrophils was observed. Cytochemical findings indicated that neutrophils and monocytes of B. orbignyanus may be involved in phagocytosis, and neutrophils play an important microbicidal role.

Effect of dietary supplementation with vitamin E and stocking density on macrophage recruitment and giant cell formation in the teleost fish, Piaractus mesopotamicus.

The effect of dietary supplementation with 0, 100 and 450 mg of vitamin E (DL-alpha tocopheryl acetate)/kg of a dry diet on the kinetics of macrophage recruitment and giant cell formation in the pacu, maintained at different stocking densities (5 kg/m(3) and 20 kg/m(3)), was investigated by insertion of round glass coverslips into the subcutaneous connective tissue. After a feeding period of 18 weeks, the coverslips were implanted and later removed for examination at 2, 7 and 15 days post-implantation. Fish fed diets supplemented with 450 mg of vitamin E showed an increase (P<0.05) in the accumulation of macrophages, foreign body giant cells and Langhans type cells. The kinetics of macrophage recruitment and giant cell formation on the glass coverslips appeared to be strongly influenced by vitamin E supplementation, since fish fed a basal diet and held at high stocking densities showed low numbers of adhering cells on the coverslips, and high concentrations of plasma corticosteroids. On the other hand, fish given a diet supplemented with 450 mg of vitamin E did not show a similar difference in plasma cortisol concentrations related to stocking density. The effect of cortisol concentrations on carbohydrate metabolism, analysed by assessment of plasma glycaemia, was not clear. Blood glucose concentrations did not vary substantially with the different treatments examined. These results suggest that vitamin E may contribute to the efficiency of the fish's inflammatory response by increasing macrophage recruitment and giant cell formation in the foreign body granulomatous reaction. Vitamin E appeared to act on the stress response of pacus by preventing a stress-related immunosuppression.

Effect of a persistent inflammatory process on experimental heterotopic ossification. The influence of macrophages.

1. We investigated the effect of a persistent carrageenin- or nystatin-induced inflammatory reaction on heterotopic ossification produced by the subcutaneous implant of a demineralized bone matrix in female Swiss mice (25 to 35 g). 2. Subcutaneous carrageenin injection (0.3 ml of a 2% solution in saline) into mice induced an inflammatory reaction characterized by a mature granuloma predominantly of macrophages containing particles of the irritant in their cytoplasm and which remained unchanged until the end of the experiment (40th day). 3. Subcutaneous nystatin inoculation (30,000 IU in 0.3 ml saline) induced an inflammatory reaction consisting initially of macrophages (4th day) but later turning into an epithelioid granuloma (7th day) consisting predominantly of epithelioid cells and which was present up to the 21st day when it was gradually replaced by adipocytes up to the 30th day. 4. An intramuscular implant of demineralized bone matrix (DBM, approximately 10 mg) induced the formation of cartilage and bone tissue and of hemopoietic bone marrow (heterotopic ossification) in 100% of the control animals (N = 5). An intramuscular DBM implant in animals that received carrageenin (N = 19) or nystatin (N = 21) induced heterotopic ossification in 100 and 57% (P < 0.01) of the animals, respectively. 5. The response to a dorsal subcutaneous DBM implant was essentially negative in control animals (N = 5), whereas implants performed near the site injected with carrageenin (N = 28) or nystatin (N = 31) produced a response in 71 (P < 0.01) and 36% (P < 0.01) of the animals, respectively. A DBM implant into the contralateral (control) dorsal subcutaneous tissue of the same animals that received carrageenin (N = 25) or nystatin (N = 29) resulted in heterotopic ossification in 64 (P < 0.01) and 7% of the animals, respectively. 6. The results suggest that the macrophages present in the mature granuloma induced by carrageenin somehow favored the development of metaplastic plates after subcutaneous DBM implant and that this effect may be systemic since the same response was observed in contralateral subcutaneous tissue.

Participation of macrophages in chloramphenicol-potentiated carrageenin-induced peritonitis in rats.

1. We investigated the possible potentiating effect of chloramphenicol succinate (30 mg/kg, every 12 h for 4 days, ip) on the response of polymorphonuclear neutrophils to carrageenin (150 micrograms, ip) or dextran (100 micrograms, ip) in the peritoneal cavity of male Wistar rats (180-230 g; N = 12 in each group). 2. Chloramphenicol potentiated the cell migration induced by carrageenin (35%) but not that induced by dextran. Previous macrophage depletion in the peritoneal cavity by washing with sterile saline abolished the cell response, whereas a previous thioglycollate-induced increase in macrophage numbers enhanced the potentiating effect (60%). 3. These results suggest that the potentiating effect on polymorphonuclear neutrophil migration induced by chloramphenicol may be related to chemotactic factors released by macrophages.

Effect of alloxan diabetes and adrenalectomy on carrageenin-induced pleurisy in the rat.

1. The effect of alloxan diabetes and adrenalectomy on the acute inflammatory response was investigated in rats using the carrageenin-induced pleurisy model. 2. Diabetic rats had reduced exudate volume (-85%) and inflammatory cell accumulation (-50%), especially polymorphonuclear leucocytes (-60%) in the pleural cavity 24 h after stimulation in relation to control values. Subcutaneous insulin administration 2 h after the inflammatory stimulus restored these parameters to normal levels. In contrast, all three measures of the response were potentiated 1.5 to 4-fold in adrenalectomized animals. 3. Results obtained by other investigators using different experimental models point to a possible pro-inflammatory function of insulin for the vascular component of inflammation. The present data show that insulin also has a significant effect on the cellular component of inflammation, especially the polymorphonuclear one, by increasing its accumulation at the affected site.

Experimental toxoplasmosis in broiler chicks.

To evaluate chicken toxoplasmosis both as an economic and a public health subject, 84 broiler chicks of a commercial strain, 30 days old, were distributed into seven groups of 12 birds (three replications of four chicks) experimentally infected with three developing T. gondii stages of the P strain as follows: tachyzoites, intravenous (two groups: 5.0 x 10(5) and 5.0 x 10(6)), cysts, per os (two groups: 1.0 x 10(2) and 1.0 x 10(3)) and oocysts, per os (three groups: 5.0 x 10(2), 5.0 x 10(3) and 5.0 x 10(4)). Twelve chicks received only a placebo (control group). During the next 30 days the following parameters were estimated: productivity (weight gain and feed conversion), clinical signs, including rectal temperature and parasitemia (bioassay). No clinical signs suggesting toxoplasmosis were seen and no statistical differences on productivity standards were found in comparison between inoculated and control chicks. However, fowls inoculated with tachyzoites and oocysts occasionally showed hyperthermia. Some haematological changes were detected in fowls inoculated with T. gondii. Anatomo-histopathological changes were not observed. From 14 parasitemias detected, 35.7% appeared on the 5th day after inoculation and 57.1% of them resulted from oocysts inoculation. After 30-35 days all birds were slaughtered: fragments from 12 organs or tissues from each of them were subjected to artificial peptic digestion and after that injected into T. gondii antibody-free mice (IIFR). T. gondii was detected in brain (12), pancreas (five), spleen (five), retina (five), kidney (two), heart (four), proventriculus (three), liver (two), intestine (two), lung (one), and skeletal muscle (one). Similar to observations with parasitemia, from 42 T. gondii isolations, 59.5% came from chicks which had received oocysts. It can thus be inferred that the developing form, expelled by cats, is the most important for T. gondii chicken infection and that brain is the most infected organ in birds. Attention must be paid to the potential importance of chicken meat in public health, since T. gondii was isolated from skeletal and heart muscles.

Experimental toxoplasmosis in pregnant bitches.

Three Toxoplasma gondii free bitches (No. 3-5) were fed 15000 sporulated T. gondii oocysts at 56, 40 and 32 day of gestation and the outcome of the pregnancy was monitored. Two of the three dogs infected during pregnancy showed evidence of congenital infection and one aborted. Two control bitches not fed oocysts delivered eight uninfected healthy pups. This study demonstrated that T. gondii can be congenitally transmitted in dogs when bitches are infected during pregnancy.

Comparative efficacy evaluation of moxidectin gel and ivermectin paste against internal parasites of equines in Brazil.

A total of 24 male and female equines of mixed breed, 10-20 months of age and naturally infected with internal parasites was utilized in a controlled test to evaluate the efficacy of a moxidectin 2% gel formulation at the dosage of 0.4 mg moxidectin per kg of live weight and an ivermectin 1.87% commercial paste formulation at the dosage 0.2 mg ivermectin per kg applied orally. Animals were allocated into three groups of eight horses each based on pre-treatment eggs per gram (EPG) counts and treatments were randomized among the groups. One group was kept as untreated controls. One animal in the moxidectin-treated group died before the end of the trial from a cause unrelated to treatment leaving a total of seven animals in this group. Fecal egg counts were performed three times post-treatment and the number of parasites remaining in each animal was determined. Statistical analyses using geometric means were performed at the 1% level of significance. Both moxidectin and ivermectin preparations reduced initial EPG from a mean of 1600 to 0 on Days 5, 7 and at the end of the trial on Day 14. Efficacy percentages of moxidectin and ivermectin against immature and adult nematodes were as follows: Trichostrongylus axei, Parascaris equorum, Strongylus edentatus, S. vulgaris, Triodontophorus spp. and Gyalocephalus capitatus, 100% for both products; Habronema muscae 99.5 and 99.6%, respectively, Strongyloides westeri, 100 and 99.2%, respectively; Oxyuris equi, 99.6 and 100%, respectively; small strongyles, 99.7% for both products. Of the latter, the most numerous were: Cylicocyclus insigne, Cylicostephanus longibursatus and Cyathostomum catinatum. No Gasterophilus nasalis were found in horses from either treated group, while two of eight control horses had infections with thisparasite. Moxidectin showed greater efficacy (84.9%) than ivermectin (67.8%) against Strongylus vulgaris larvae found in the mesenteric artery aneurisms, but the difference was not statistically significant. Total parasite counts for both treated groups were significantly lower (p<0.01) than in the non-treated group. No significant differences were noted between moxidectin and ivermectin. Efficacy against the 30 nematode species found in this study was very evident for both products. As expected, neither moxidectin nor ivermectin was effective in controlling the tapeworm Anoplocephala perfoliata. No adverse reactions were observed during the experimental period.

Alternative treatment for Anacanthorus penilabiatus (Monogenea: Dactylogyridae) infection in cultivated pacu, Piaractus mesopotamicus (Osteichthyes: Characidae) in Brazil and its haematological effects.

The present work, studied the effect of 0, 1,000, 1,500 and 2,000 mg of garlic powder/kg dry ration for Piaractus mesopotamicus (Osteichthyes: Characidae), weighting 73.6 +/- 39.4 g and measuring 15.0 +/- 2.7 cm, fed for a period of 15, 30 and 45 days. Fifteen days after treatment with 1,000 and 2,000 mg of garlic/kg dry ration, significant reduction of Anacanthorus penilabiatus (Monogenea: Dactylogyridae) in the gills was related. Nevertheless, the addition of garlic to the ration caused significant increase in the erythrocyte number and in the thrombocyte percentage in the circulating blood. However, a decrease in the lymphocyte percentage was also observed. After 45 days, fish fed with garlic showed significant increase in the erythrocyte number, leucocyte, haemoglobin rate, hematocrit and thrombocyte.

Piscinoodinium pillulare (Schäperclaus, 1954) Lom, 1981 (Dinoflagellida) infection in cultivated freshwater fish from the northeast region of Sao Paulo State, Brazil. Parasitological and pathological aspects.

The Aquaculture Center of Unesp, Jaboticabal, SP, Brazil, received fishes for diagnosis from fish farmers reporting fish crowding at pond edges and in water inlets. Fifty-three out of 194 cases showed round to oval, immobile whitish structures, measuring up to 162 microm diameter, identified as the dinoflagellate Piscinoodinium pillulare. In 34 diagnosed cases the parasites were present in the gills, in 2 on body surface, and 9 in both. Thirty-one out of 53 were tambacu hybrids hosts; 7, Piaractus mesopotamicus; 6, Colossoma macropomum; 5, Leporinus macrocephalus; 3, Oreochromis niloticus; and 1, Prochilodus lineatus. Fish showed increased mucous production on body surface and gills, while ecchymosis in the caudal peduncle and operculum was present. The gills also showed paleness, congestion, and petechiae. Histopathology presented a great number of trophonts situated between secondary lamellae, fixed to or detached from the epithelium. Primary lamellae presented interstitial hemorrhages, severe hyperplasia of the epithelium, goblet cells, and mononuclear infiltrates. The present work is the first report of P pillulare in Brazil and emphasizes the importance of this dinoflagellate which caused significant economic losses from 1995 through 1997.

Prevalence and histopathology of Neoechinorhynchus curemai Noronha, 1973 (Acanthocephala: Neoechinorhynchidae) in Prochilodus lineatus Valenciennes, 1836 from Volta Grande Reservoir, MG, Brazil.

The present work studied the prevalence and histopathology of Neoechinorhynchus curemai Noronha, 1973 (Acanthocephala; Neoechinorhynchidae) from curimbatá, Prochilodus lineatus Valenciennes, 1836. Eighteen fishes with averages of 46.7 +/- 1.1 cm length and 1,674.8 +/- 75.6 g weight were collected with net, bimonthly from December 1995 thru December 1996 in the hydroelectric power station of Volta Grande Reservoir (Cemig), Minas Gerais, Brazil. From analysed fishes, 15 were infected with acanthocephalans in the intestine (prevalence 83.3%). The greatest mean intensity occurred in August 1996 with 66.5 (16 to 208) parasites. Histopathological analysis showed complete desquamation of the intestinal epithelium with severe hyperplasia and hypertrofia of the goblet cells. Severe inflammatory reaction at the submucosa, displacement of their sheaf, associated with oedema and mononuclear and eosinophilic infiltration were observed.

Deformities in reared cobia, Rachycentron canadum L. and grouper, Epinephelus marginatus, in São Paulo state coast, Brazil: case report / Deformidades em cobias, Rachycentron canadum L, e garoupas, Epinephelus marginatus, cultivadas na costa do estado de São Paulo, Brasil: relato de caso

Skeletal deformities are frequent problems in cultured fish populations due mainly to unfavorable abiotic conditions, inappropriate nutrition, and genetic factors. These may cause erratic swimming, decrease of conversion rate, growth, and market value. The aim of this case report was to present the occurrence of deformities in juvenile cobia and groupers from an offshore marine farm in Ilhabela, São Paulo, Brazil. Nine cobias and seven juvenile groupers, that presented deformities, were euthanized and fixed in 10% formaldehyde. They were tagged, processed, and stained with "Alcian Blue" (AB) and "Alizarin Red-S" (ARS) for visualization of cartilage and bone tissue, respectively. After evisceration, radiographic examination was performed. The deformities observed in cobia were atrophy of the operculum, disorganization of the gill lamellae, deformity of the lower jaw, kyphosis, and scoliosis. Groupers presented lower jaw deformity and saddleback syndrome. The farm studied in this case report had poor management standards, fed commercial feed for freshwater herbivorous fish and cooked sardines, high inbreeding and absence of adequately trained professionals to manage these species. The sum of these factors must have directly influenced the appearance of these deformities. It is necessary to control this situation to minimize the incidence of malformations that have economic impacts to farmers.(AU)

As deformidades esqueléticas são problemas frequentes em populações de peixes cultivados devido, principalmente, a condições abióticas desfavoráveis, nutrição inapropriada e fatores genéticos. Estas podem causar natação errática, diminuição do índice de conversão, do crescimento e do valor no mercado. O objetivo deste relato de caso foi apresentar a ocorrência de deformidades em cobias e garoupas juvenis provenientes de uma fazenda marinha em sistema offshore, em Ilhabela, São Paulo, Brasil. Nove cobias e sete garoupas juvenis, que apresentavam deformidades, foram sujeitos à eutanásia e fixados em formol a 10%. Os espécimes foram registrados, processados e corados com "Alcian Blue" (AB) e "Alizarin Red-S" (ARS) para visualização de tecido cartilaginoso e ósseo, respectivamente. Os peixes foram eviscerados para o exame radiográfico. As deformidades observadas em cobia foram: atrofia do opérculo, desorganização das lamelas branquiais, deformidade do maxilar inferior, cifose e escoliose. As garoupas apresentaram deformidade do maxilar inferior e síndrome de Saddleback. Os peixes deste relato de caso estiveram sob manejo deficiente, alimentados com ração comercial para peixes herbívoros de água doce e sardinhas cozinhadas, alta consanguinidade e ausência de profissionais adequadamente treinados para o manejo dessas espécies. A soma desses fatores influenciou diretamente na aparição dessas deformidades, precisando ser controlados para minimizar a incidência de malformações que tenham impacto econômico para os produtores.(AU)

Ativação de células de memória na produção de anticorpos e na expressão de células IgM positivas no baço de tilápias-do-nilo / Activation of memory cells in the antibody production and the expression of IgM positive cells in Nile tilapia spleen

O presente trabalho avaliou o papel do baço no armazenamento e na reativação das linhagens de células B, representadas por células IgM positivas imunomarcadas no tecido esplênico, bem como a funcionalidade dessas células, sobre a cinética dos linfócitos e na produção sistêmica de anticorpos em tilápias-do-nilo (Oreochromis niloticus). Foram separados dois grupos: grupo memória, constituído por peixes previamente imunizados com hemácia de carneiro a 2,5%, para a geração da memória imune, e o grupo naive, que recebeu o mesmo volume de solução salina a 0,65%. Após 32 dias, os dois grupos foram submetidos a uma nova dose do antígeno na mesma concentração, volume e via de inoculação. A reativação dos clones de memória foi evidenciada pelo aumento do número de células IgM positivas no baço do grupo memória no dia zero/pré-imune. Além disso, o mesmo grupo apresentou aumento dos títulos de anticorpos séricos no 14º dia e no número absoluto de linfócitos no 21º dia em relação ao grupo naive. Esses resultados sugerem que o baço não seja apenas um local de armazenamento, mas também de reativação de células B de memória em tilápia-do-nilo.(AU)

This work aimed to evaluate the role of the spleen in storage and reactivation of the memory B cells, represented by IgM positive cells and the systemic production of sheep antibodies anti-red cell in Nile tilapia (Oreochromis niloticus). Two groups were established: the memory group, containing fish previously immunized with a 2,5% sheep anti-red cell, to generate the immune memory; and the naive group, containing fish that received a 0,65% saline solution. After 32 days, both groups were subjected to a new dose of the same antigen at the same concentration, volume, and inoculation via. The memory clones reactivation was correlated to the increase of the IgM positive cells in the spleen in the memory group at 0 day. The memory group showed an increase in the absolute number of lymphocytes at 21 days and an increase in the antibodies at 14 days after inoculation when compared to the naive group. The results suggest that the spleen may be a storage and reactivation place of memory B cells in Nile tilapia.(AU)

Transferência passiva de soro hiperimune anti-Streptococcus agalactiae e seu efeito profilático em tilápias-do-nilo infectadas experimentalmente: sobrevivência e títulos de anticorpos / Passive transfer of hyperimmune serum anti Streptococcus agalactiae and its prophylactic effect on Nile tilapia experimentally infected

A bactéria Streptococcus agalactiae é um potente agente causador de surtos por doenças bacterianas em peixes. O estresse provocado pelo manejo zootécnico e pela má qualidade ambiental torna a tilápia susceptível às infecções por essa bactéria. O objetivo do presente trabalho foi avaliar a resistência de tilápias-do-nilo imunizadas com soro hiperimune anti-S. agalactiae, posteriormente desafiadas com cepa homóloga da mesma bactéria. Após determinação da DL 50 de S. agalactiae, 36 tilápias foram distribuídas em quatro aquários, dois para o grupo controle e dois para inoculação celomática para produção de anticorpos anti-S. agalactiae. No 21° e 28° dias, foi coletado sangue para obtenção de soro hiperimune utilizado na transferência passiva. Em seguida, 30 tilápias foram distribuídas em três aquários e submetidas a três tratamentos: GI: controle; GII: imunizadas com o soro inativado; GIII: imunizadas com soro ativo...

The Streptococcus agalactiae bacteria is a potent agent which causes outbreaks of bacterial diseases in fish. The stress caused by management and poor environmental quality makes tilapia susceptible to infections, including by bacterium. The aim of this study was to evaluate the resistance of the Nile tilapia immunized with hyperimmune serum against S. agalactiae subsequently challenged with homologous strain of the same bacteria. After determining the DL 50 of S. agalactiae, 36 tilapias were distributed in 4 aquariums, 2 for the control group and 2 for the group via coelomic, inoculated with the DL 50 for anti-S. agalactiae antibodies production. On the 21st and 28th day blood was collected for the obtainment of hiperimmune serum used in passive transference. Then, 30 tilapias were distributed in 3 aquariums submitted to 3 treatments (GI: control; GII: immunized with inactivated-serum; GIII: immunized with non-inactivated serum)...