Imunidade celular em vacas Holandesas soropositivas para o Vírus da Leucose Bovina (BLV) durante o período de transição

ABSTRACT This research evaluated the dynamics of leukocytes and lymphocytes subsets in seropositive Holstein cows for BLV during the transition period. Blood samples (n=72) from 12 cows were harvested from week -2 up to week +3 to perform leukogram, immunophenotyping, cortisol and haptoglobin (Hp). Leukocytes pattern was characterized by leukocytosis, neutrophilia, monocytosis and eosinopenia around calving. Lymphocytosis and high proportions of B cells CD21+ were a constant finding between week -2 and +3, thus cows were tested and confirmed seropositive for BLV. The values of T lymphocytes subsets were low during the transition period, observing two peaks that coincided with high levels of cortisol at delivery (2.11µg/dL) and week +3 (1.97µg/dL). Hp had gradual increase from week -2 (166µg/mL) until week +3 (576g/mL) probably due to high rate of uterine infection detected between week +2 and +3. The seropositive cows for BLV presented stress leukogram around delivery, except for lymphocytes. Lymphocytosis and the high proportions of B cells, associated with the low proportions of T lymphocytes, can be indicative of immunosuppression and predisposition to the inflammatory process observed in the post-partum period.

RESUMO Esta pesquisa avaliou a dinâmica dos leucócitos e das subpopulações de linfócitos em vacas Holandesas soropositivas para o BLV no período de transição. Amostras de sangue (n=72) provenientes de 12 vacas foram coletadas entre as semanas -2 e +3 para a realização do leucograma, imunofenotipagem, dosagem de cortisol e haptoglobina (Hp). O perfil leucocitário foi caracterizado por leucocitose, neutrofilia, monocitose e eosinopenia próximo ao parto. Linfocitose e elevada proporção de linfócitos B CD21+ foram achados constantes entre as semanas -2 e +3; assim, as vacas foram testadas e confirmadas soropositivas para o BLV. Os valores das subpopulações de linfócitos T apresentaram-se baixos durante o período de transição, observando-se dois picos máximos que coincidiram com as elevações nas concentrações de cortisol no parto (2,11µg/dL) e semana +3 (1,97µg/dL). Hp apresentou aumento crescente de -2 (166µg/mL) a +3 (576µg/mL), provavelmente associada à elevada taxa de infecções uterinas observadas nas semanas +2 e +3. As vacas soropositivas para o BLV apresentaram leucograma de estresse próximo ao parto, exceto para linfócitos. A linfocitose e as elevadas proporções de células B CD21+, associadas com as baixas proporções de células T, podem ser indicativo de imunossupressão e predisposição aos processos inflamatórios no período pós-parto.

Experimental infection of commercial layers with wild or attenuated Salmonella Gallinarum mutant strains: anatomic pathology, total blood cell count and serum protein levels

The aim of the present study was to comparatively evaluate hemogram, blood serum components and anatomopathologic alterations in commercial layers experimentally challenged with an attenuated vaccine candidate strain (SGcobScbiA) and other two pathogenic strains (SGDcobS and SGNalr) of Gallinarum (SG). In total, 280 commercial layers were randomly divided into 4 groups (G1, G2, G3 and G4). At five days of age, birds from groups G1 received approximately 107 colony forming units (CFU) of SGDcobS; meanwhile birds from group G2 and G3 received the same dose of SGNalr and SGcobScbiA, respectively. Birds from G4 were not infected. At 24 hours before (DBI) and 24 hours after (1 DAI), and three (3 DAI), five (5 DAI), seven (7 DAI) ten (10 DAI), and fifteen (15 DAI) days after the infection, 10 birds of each group were humanely killed and blood samples collected to hematological and serum tests. Samples of liver, spleen, thymus, bursa of Fabricius, kidney and heart were also collected for the histological examination. Birds inoculated with SGDcobS and SGNalr showed similar alterations in hemogram, blood serum components and anatomopathologic exams. On the other hand, the exams of birds inoculated with SGcobScbiA strain were similar to those of the uninfected birds. However, changes could be noticed in levels of uric acid and cholesterol during the course of the infection of birds from G3. Decrease in levels of light IgG 3 DAI was also observed in birds from this group. Pyknosis in kidney cells was a microscopic alteration found in birds from G3. Further studies must be done to verify if these alterations will not interfere in the performance of the vaccinate birds with SGcobScbiA strain.

Experimental infection of commercial layers using a Salmonella enterica serovar Gallinarum strain: Leukogram and serum acute-phase protein concentrations

The aim of the present study was to evaluate white blood cell counts and serum protein profiles of commercial layers experimentally infected with Salmonella Gallinarum (SG) in order to better understand the pathophysiology of the disease caused by this bacterium. 180 five-day-old commercial layers were divided into 3 groups (G); G1 and G2 received 0.2 mL of inoculate containing 3.3x10(8) CFU or 3.3x10(5) CFU SG resistant to nalidix acid (Nal r)/mL, respectively, directly into their crops. G3 group did not receive the inoculum. Birds were sacrificed 24 hours before (T1) and 24 hours after the infection (T2), and three (T3), five (T4), seven (T5), and ten (T6) days after the administration of the inoculum. White blood cell counts were carried out in a Neubauer hemocytometer and in blood smears. Serum protein concentrations, including acute-phase proteins, were determined using sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE). Data were submitted to analysis of variance, and means were compared by Tukey's test (P 0.05). G1 and G2 groups presented higher leukocyte counts on T4 and T5, respectively, due to the increase of circulating lymphocytes and heterophils, with a significant difference relative to G3. In electrophoresis, an increase in the serum levels of ceruloplasmin, haptoglobin, and hemopexin and a decrease in transferrin, which are acute-phase proteins, was verified. IgA serum levels did not change; however, IgG concentration increased during the infection. In conclusion, the results provide information for the better understanding of the pathophysiology of fowl typhoid.

Experimental infection of commercial layers using a Salmonella enterica serovar Gallinarum strain: Leukogram and serum acute-phase protein concentrations

The aim of the present study was to evaluate white blood cell counts and serum protein profiles of commercial layers experimentally infected with Salmonella Gallinarum (SG) in order to better understand the pathophysiology of the disease caused by this bacterium. 180 five-day-old commercial layers were divided into 3 groups (G); G1 and G2 received 0.2 mL of inoculate containing 3.3x10(8) CFU or 3.3x10(5) CFU SG resistant to nalidix acid (Nal r)/mL, respectively, directly into their crops. G3 group did not receive the inoculum. Birds were sacrificed 24 hours before (T1) and 24 hours after the infection (T2), and three (T3), five (T4), seven (T5), and ten (T6) days after the administration of the inoculum. White blood cell counts were carried out in a Neubauer hemocytometer and in blood smears. Serum protein concentrations, including acute-phase proteins, were determined using sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE). Data were submitted to analysis of variance, and means were compared by Tukey's test (P 0.05). G1 and G2 groups presented higher leukocyte counts on T4 and T5, respectively, due to the increase of circulating lymphocytes and heterophils, with a significant difference relative to G3. In electrophoresis, an increase in the serum levels of ceruloplasmin, haptoglobin, and hemopexin and a decrease in transferrin, which are acute-phase proteins, was verified. IgA serum levels did not change; however, IgG concentration increased during the infection. In conclusion, the results provide information for the better understanding of the pathophysiology of fowl typhoid.

Blood serum components and serum protein test of Hybro-PG broilers of different ages

Blood serum samples of HYBRO PG broilers were analyzed, with 30 samples collected from 21-day-old broilers (G1), 30 from 35-day-old birds (G2), and 30 from 42-day-old birds (G3), with the aim of establishing normal values of some blood serum parameters. The activities of the enzymes gamma-glutamyl-transferase (GGT), aspartate aminotransferase (AST), creatine kinase (CK), alkaline phosphatase (ALP), and lactate dehydrogenase (LDH), serum levels of total calcium, calcium ion, phosphorus, sodium, potassium, magnesium, chlorides, creatinine, uric acid, triglycerides, cholesterol, total protein, albumin, total and indirect and direct bilirubin, and electrophoretic profile of serum proteins in acrylamide (SDS-PAGE) and agarose gel were determined. There was no influence of age on total bilirubin and albumin levels. All the other evaluated parameters presented differences in at least one age group. Protein electrophoretic profile also changed as a function of age. The obtained results can be considered as normal for the studied ages, and therefore be used as references for the interpretation of laboratory exams of broilers of this genetic line in the evaluated ages.

Infection of commercial laying hens with Salmonella Gallinarum: clinical, anatomopathological and haematological studies

This study aimed at evaluating the susceptibility of commercial laying hens to Salmonella Gallinarum (SG). Two experiments were carried using a mutant strain of Salmonella Gallinarum resistant to nalidix acid (SGNALr). In the first trial, the resistance of birds was evaluated based on clinical signs, faecal shedding, and mortality. It was carried out with six lines of commercial layers being three light white layers, considered to be resistant to SG (W1, W2, W3), and three semi-heavy brown varieties (B1, B2, B3), considered susceptible to SG. Each group contained 15 one-day-old birds. Hens were inoculated in the crop at 5 days of age with 0.2 mL of SGNALr neat culture. In addition, to each brown variety, a new group of 15 birds was challenged with 0.2mL of the same SGNALr culture diluted at 10-3. At the end of the first experiment, the surviving birds were sacrificed, and microbiological culture of liver and spleen was performed. In the second experiment, white and brown birds were inoculated with neat culture at five days of age. Samples were collected for evaluation of blood parameters and histopathology assessment at 1, 3, 5, 7, 9, 12, and 14 days post-infection. The results of the first experiment showed higher resistance of white birds (p 0.05), although there was no uniformity in the responses against fowl typhoid among the birds within these groups. In the second experiment, there were differences between white and brown birds both in blood parameters and in organ lesion intensity.

Infection of commercial laying hens with Salmonella Gallinarum: clinical, anatomopathological and haematological studies

This study aimed at evaluating the susceptibility of commercial laying hens to Salmonella Gallinarum (SG). Two experiments were carried using a mutant strain of Salmonella Gallinarum resistant to nalidix acid (SGNALr). In the first trial, the resistance of birds was evaluated based on clinical signs, faecal shedding, and mortality. It was carried out with six lines of commercial layers being three light white layers, considered to be resistant to SG (W1, W2, W3), and three semi-heavy brown varieties (B1, B2, B3), considered susceptible to SG. Each group contained 15 one-day-old birds. Hens were inoculated in the crop at 5 days of age with 0.2 mL of SGNALr neat culture. In addition, to each brown variety, a new group of 15 birds was challenged with 0.2mL of the same SGNALr culture diluted at 10-3. At the end of the first experiment, the surviving birds were sacrificed, and microbiological culture of liver and spleen was performed. In the second experiment, white and brown birds were inoculated with neat culture at five days of age. Samples were collected for evaluation of blood parameters and histopathology assessment at 1, 3, 5, 7, 9, 12, and 14 days post-infection. The results of the first experiment showed higher resistance of white birds (p 0.05), although there was no uniformity in the responses against fowl typhoid among the birds within these groups. In the second experiment, there were differences between white and brown birds both in blood parameters and in organ lesion intensity.

Blood serum components and serum protein test of Hybro-PG broilers of different ages

Blood serum samples of HYBRO PG broilers were analyzed, with 30 samples collected from 21-day-old broilers (G1), 30 from 35-day-old birds (G2), and 30 from 42-day-old birds (G3), with the aim of establishing normal values of some blood serum parameters. The activities of the enzymes gamma-glutamyl-transferase (GGT), aspartate aminotransferase (AST), creatine kinase (CK), alkaline phosphatase (ALP), and lactate dehydrogenase (LDH), serum levels of total calcium, calcium ion, phosphorus, sodium, potassium, magnesium, chlorides, creatinine, uric acid, triglycerides, cholesterol, total protein, albumin, total and indirect and direct bilirubin, and electrophoretic profile of serum proteins in acrylamide (SDS-PAGE) and agarose gel were determined. There was no influence of age on total bilirubin and albumin levels. All the other evaluated parameters presented differences in at least one age group. Protein electrophoretic profile also changed as a function of age. The obtained results can be considered as normal for the studied ages, and therefore be used as references for the interpretation of laboratory exams of broilers of this genetic line in the evaluated ages.

Hemograma e proteinograma plasmático de eqüinos hígidos e de eqüinos acometidos por abdômem agudo, antes e após laparotomia

Hemogram and plasma protein concentrations of healthy horses and horses affected with acute abdomen before and after laparotomy were determined to investigate if these determinations can be of help on the diagnosis and prognosis of the post-operative intercurrence. The body temperature, respiratory and heart rates, red blood cell, leukocyte and neutrophil counts, packed cell volume, and plasma protein concentrations increased after laparotomy, mainly in sick horses. Acute phase protein concentrations were higher in the horses affected with acute abdomen than in the healthy horses with the highest values detected at about 48 hours after surgery. The results suggest that determining the level of these proteins can be useful for the diagnosis and prognosis of post-operative recovery after laparotomy in healthy horses and in horses affected with acute abdomen.

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.

Ocorrência de poliúria-polidpsia em eqüinos estabulados - descrição de dois casos de polidipsia psicogênica

Two four-year-old male Brasileiro de Hipismo breed horses presenting signs of polyuria-polydipsia, were submitted to a complete clinical evaluation as well as having blood and urine samples collected for laboratory analysis. No clinical, hematological or biochemical abnormalities were observed, but a significant decrease in urine specific gravity was detected in both horses. One of the horses presented 1,009 and the other 1,008. A 24-hour water deprivation test was carried out and, during this period, the urine specific gravity gradually increased reaching the normal values of 1,028 and 1,026. The abnormal behavior occured as a consequence of an excessive confinement. The clinical and laboratory results associated to the 24-hour water deprivation test indicated a diagnosis of psychogenic polydipsia.

Dois eqüinos da raça Brasileira de Hipismo, machos, quatro anos de idade, apresentando sinais de poliúria-polidipsia, foram submetidos a completa avaliação clínica e a exames de sangue e urina. Não foram observadas alterações clínicas, hematológicas ou bioquímicas, a não ser um decréscimo na gravidade específica da urina de ambos os animais (1,009 e 1,008). Realizou-se um teste de privação de água de 24 horas e durante esse período a gravidade especifica da urina subiu gradualmente até atingir a normalidade (1,028 e 1,026, respectivamente). O comportamento anormal ocorreu por confinamento excessivo. Os resultados clínicos e de laboratório e o teste de privação de água indicam o diagnóstico de polidipsia psicogênica.