Anemia in canine chronic kidney disease is multifactorial and associated with decreased erythroid precursor cells, gastrointestinal bleeding, and systemic inflammation.

OBJECTIVE: Compare erythropoiesis-related factors between different stages of canine chronic kidney disease (CKD). ANIMALS: 8 healthy adult dogs (controls), and 24 dogs with CKD, equally divided into 3 groups based on International Renal Interest Society-CKD Guidelines (stage 2, 3, and 4) were recruited between December 2012 and December 2014. METHODS: The following were assessed in all dogs and then compared between groups: bone marrow cytology, CBC, reticulocyte count, urinalysis, serum biochemistry, blood pressure, occult gastrointestinal bleeding, and serum concentrations of parathyroid hormone (PTH), erythropoietin, interleukin-1ß, interleukin-3, tumor necrosis factor-α (TNFα), and interferon-γ. RESULTS: Erythropoiesis inducing and suppressing factors and the results of the bone marrow cytology of dogs in stage 2 CKD did not differ from the control group. The presence of reticulocytosis in CKD stage 2 suggests that blood loss or erythrocyte destruction might be contributing to developing anemia. Anemia in dogs with progressive CKD was associated with increasing PTH and TNFα and with elevation of the ratio of myeloid to erythroid precursor cells caused by hypoplasia of the erythroid series. The latter was represented mainly by a decrease in the population of polychromatophilic rubricytes and metarubricytes. CLINICAL RELEVANCE: Increased PTH and TNFα seem to contribute to the reduced percentage of polychromatophilic rubricytes and erythroid population, thereby aggravating the anemia of dogs with advanced CKD. Gastrointestinal blood loss contributes to anemia in all canine CKD stages.

Serum proteinogram and biochemistry of Holstein cows in the peripartum period / Proteinograma e bioquímica séricas de vacas da raça Holandesa no período periparto

In face of the few reports found in national literature analyzing the potential influence of parturition number in serum proteinogram and biochemical profile in the peripartum period of high yielding dairy cows, the aim of the present study was to comparatively evaluate the dynamics of these serum constituents' concentrations in blood samples obtained from primiparous and multiparous Holstein cows, 60 and 30 days prepartum and in the day of parturition. Data were analyzed by repeated measures variance analysis (ANOVA) and differences between groups and moments were analyzed by Tukey's test. Results were considered significant when P<0.05. Parity influenced levels of total protein, albumin, globulins, magnesium, cholesterol, which were higher in multiparous cows, as well as concentrations of ceruloplasmin, total calcium, chloride and alkaline phosphatase activity, which were higher in primiparous cows. Parturition influenced serum concentrations of ceruloplasmin (+58%), transferrin (-25%), haptoglobin (+33%), total protein (-17%), globulins (-25%), immunoglobulin A (-43%), immunoglobulin G (-24%), total calcium (-12%), inorganic phosphorus (-10%), chloride (+5%), sodium (+4%), cholesterol (-23%), triglycerides (-38.6%), as well as activities of aspartate aminotransferase (+14%) and alkaline phosphatase (+28%). A decrease in serum levels of total calcium, inorganic phosphorus, cholesterol and triglycerides was more pronounced in multiparous than in primiparous cows. These results demonstrate that the interpretation of proteinogram and serum constituents should take into consideration lactation number and the moment of parturition as relevant factors in high yielding dairy cows in the transition period.(AU)

Diante da escassez de relatos encontrados na literatura nacional quanto à potencial influência do número de parições sobre o proteinograma sérico e perfil bioquímico no período periparto de vacas leiteiras de alta produção, o objetivo do presente estudo foi avaliar comparativamente a dinâmica de constituintes séricos em amostras de sangue obtidas de vacas da raça Holandesa primíparas e pluríparas, 60 e 30 dias pré-parto e no dia do parto. Os resultados foram avaliados por análise de variância (ANOVA) com medidas repetidas no tempo e as diferenças entre grupos e entre momentos foram analisadas pelo teste de Tukey, sendo os resultados considerados significativos quando P<0,05. O número de parições influenciou os teores de proteína total, albumina, globulinas, magnésio e colesterol, que foram maiores em vacas pluríparas, bem como as concentrações de ceruloplasmina, cálcio total, cloreto e atividade de fosfatase alcalina, que foram maiores em vacas primíparas. O número de parições influenciou as concentrações séricas de ceruloplasmina (+58%), transferrina (-25%), haptoglobina (+33%), proteína total (-17%), globulinas (-25%), imunoglobulina A (-43%), imunoglobulina G (-24%), cálcio total (-12%), fósforo (-10%), cloretos (+5%), sódio (+4%), colesterol (-23%), triglicérides (-38.6%), bem como as atividades de aspartato aminotransferase (+14%) e fosfatase alcalina (+28%). A diminuição do teor sérico de cálcio total, fósforo, colesterol e triglicérides foi mais acentuada em vacas pluríparas do que em vacas primíparas. Esses resultados mostram que a interpretação do proteinograma e dos constituintes séricos deve levar em consideração o número de lactações e a ocorrência do parto como fatores relevantes em vacas leiteiras de alta produção no período de transição.(AU)

Blood serum acute phase proteins and iron dynamics during acute phase response of Salmonella enterica serotype Dublin experimentally infected buffalo calves.

The study aimed to evaluate clinical signs, blood serum acute phase proteins (APP) and iron dynamics during the acute phase response (APR) of Salmonella Dublin experimentally infected Murrah buffalo calves. Six buffalo calves constituted the control group (CNT) and six were orally inoculate with 108 CFU of S. Dublin (INF). Clinical evaluation was performed, rectal swabs to detect S. Dublin strains were collected and venous blood was sampled before and throughout seven days after inoculation. The APP fractions ß-haptoglobin, α-haptoglobin, ceruloplasmin and transferrin were analyzed by 1-D and 2-D electrophoresis. Proteins were identified using LC/ESI-MS/MS and NCBI database. Plasma fibrinogen, serum iron and serum haptoglobin concentrations were measured. The inoculation of 108 CFU of S. Dublin was effective in inducing clinical signs of Salmonellosis, such as hyperthermia and diarrhea. 1-DE showed that ß and α-haptoglobin increased 204% (p = 0.008) and 184% (p = 0.022) 48 h after inoculation (HAI), respectively, with highest concentrations 120 HAI (498% increased, p = 0.012; 431% increased, p = 0.011) and 168 HAI (492% increased, p = 0.019; 523% increased, p = 0.028). 2-DE showed that the expression of two spots, identified as ß-haptoglobin, were increased 693% (p = 0.0006) and 580% (p = 0.0003) 168 HAI, respectively, while one spot, identified as α-haptoglobin, increased 714% (p = 0.040). Haptoglobin concentrations increased 1339% (p < 0.0001) 168 HAI. 1-DE showed that ceruloplasmin increased 42% (p = 0.034) 48 HAI, with highest concentration 120 HAI (133% increased, p = 0.022). 2-DE showed that the expression of two spots, identified as ceruloplasmin, were increased 218% (p = 0.0153) and 85% (p = 0.0143) 168 HAI, respectively. Fibrinogen increased 78% (p = 0.012) 96 HAI, with highest concentration 120 HAI (increased 114%, p = 0.002). Iron decreased 33% 24 HAI (p = 0.015) and 37% 72 HAI (p = 0.029), and began to be restored 96 HAI. 1-DE showed that transferrin decreased 23% 120 HAI (p = 0.047), and that values were restored 168 HAI. 2-DE showed that expression patterns of transferrin comparing 0 h and 168 HAI were similar, evidencing that values were restored 168 HAI. In conclusion, the inoculation of 108 CFU was effective in inducing hyperthermia and diahrrea. ß and α-haptoglobin, ceruloplasmin and fibrinogen worked as positive APP during the APR to S. Dublin infection and are potential biomarker candidates. Concentrations of iron and transferrin decreased during the infection, highlighting the fact that mechanisms for restricting iron availability are part of the APR triggered against S. Dublin infection in buffalo calves.

Reference 1D and 2D electrophoresis maps for potential disease related proteins in milk whey from lactating buffaloes and blood serum from buffalo calves (Water buffalo, Bubalus bubalis).

The aim of this study was to identify potential disease related proteins in milk whey of lactating buffaloes and blood serum of buffalo calves, in order to define a reference electrophoresis map for 1-DE and 2-DE. Additionally, changes in some protein patterns from buffalo calves during salmonellosis and lactating buffaloes during mastitis are presented. Milk samples were collected and distributed into groups: Milk samples from healthy buffaloes (SCC < 100.000 cells/ml, negative microbiology and CMT) (G1, n = 5) and buffaloes with subclinical mastitis (SCC > 500.000 cells/ml, positive microbiology and CMT) (G2, n = 5). Blood samples from buffalo calves (n = 6) were collected, and three calves were experimentally infected with Salmonella Dublin and samples analyzed before (M0) and 72 h after inoculation (M1). 1-DE was accomplished by loading 10 µg of TP into SDS-PAGE, stained with Coomassie blue. 2-DE was accomplished by loading 200 µg of TP into 11 cm, pH 3-10 non-linear IPG strips, followed by SDS-PAGE, stained with Coomassie blue. Protein bands/spots were excised, subjected to tryptic in-gel digestion and analyzed by LC/ESI-MS/MS. Protein identity was assigned using NCBI databases. After bands/spots from 1-DE and 2-DE were analyzed, a protein map with 35 and 40 different identified proteins in blood serum and milk whey, respectively, was generated. Significant changes in patterns of haptoglobin were observed in buffalo calves with salmonellosis and in patterns of IgLC, ß-lactoglobulin and α-lactalbumin of lactating buffaloes during mastitis. The establishment of a protein map for 1-DE and 2-DE, identifying potential disease related proteins, can help to address alterations during diseases in buffaloes.

Blood gas and serum biochemical RIs for healthy newborn Murrah buffaloes (Bubalus bubalis).

BACKGROUND: There is a lack of published work on RIs for newborn buffaloes. Establishing blood gas and serum biochemical RIs for newborn buffaloes is important for monitoring health. OBJECTIVES: This study establishes blood gas and serum biochemical RIs of newborn buffaloes. METHODS: Twenty-eight newborn buffaloes, 10-30 days old, were selected. Thirty blood biochemical variables were analyzed. The Anderson-Darling test was used to assess the normality of the distribution. The Dixon test and the Tukey test were used to identify outliers. The RI and 90% CI were determined using standard and robust methods and the Box-Cox transformation. RESULTS: A total of 30 RIs for healthy buffalo calves have been reported in this study. RIs for blood gas variables were reported for pH, partial pressure of oxygen (pO2 ), partial pressure of carbon dioxide (pCO2 ), saturation of O2 (SO2 ), bicarbonate (cHCO3- ), base excess (BE), total carbon dioxide (ctCO2 ), and anion gap (AG). RIs for serum biochemical variables were reported for glucose (GLU), direct bilirubin (DB), total bilirubin (TB), AST, ALP, GGT, CK, LDH, creatinine (CREA), urea, cholesterol (CHOL), triglycerides (TG), Ca, P, Mg, Na, K, iCa, Cl, iron, total protein (TP), and albumin (ALB). CONCLUSIONS: This is the first reported study covering complete serum chemistry and blood gas RIs for healthy 1-month-old Murrah buffaloes.

Perfil bioquímico, inclusive proteinograma, do soro lácteo de búfalas primíparas e pluríparas sadias ao longo da lactação / Biochemical profile, including protein concentration, of whey from healthy primiparous and pluriparous buffaloes during lactation

Para avaliar o perfil bioquímico, inclusive proteínas, do soro lácteo de búfalas Murrah primíparas e pluríparas sadias foram analisadas amostras de leite de 30 fêmeas bubalinas durante uma lactação completa. Os animais foram distribuídos em três grupos: G1 - 10 búfalas primíparas, G2 - 10 búfalas pluríparas com duas a três lactações e G3 - 10 búfalas pluríparas com mais de três lactações. O período de lactação foi dividido em: fase inicial (I: primeiro ao terceiro mês de lactação), fase intermediária (T: quarto ao sexto mês de lactação) e fase final (F: sétimo ao nono mês de lactação). Antes da colheita das amostras de leite foram realizados o exame físico da glândula mamária, o teste da caneca de fundo escuro e o California Mastitis Test (CMT). Após a assepsia dos quartos mamários, foram colhidas mensalmente, durante uma lactação completa, amostras de 20mL de leite de cada quarto mamário, em frascos plásticos esterilizados e sem conservante, para a realização do isolamento microbiológico, determinação do perfil bioquímico e fracionamento proteico por meio de eletroforese em gel de poliacrilamida contendo dodecil sulfato de sódio (SDS-PAGE), e amostras de 30mL de leite de cada quarto mamário, em frascos plásticos esterilizados contendo conservante bronopol, para contagem de células somáticas (CCS). Das 1.042 amostras de leite colhidas dos três grupos experimentais durante a lactação, 923 amostras de leite apresentaram reação negativa ao CMT e isolamento microbiológico negativo e foram selecionadas para as análises do perfil bioquímico e fracionamento proteico em SDS-PAGE. Notou-se influência da ordem de parto e da fase da lactação no perfil bioquímico e no proteinograma do soro lácteo de búfalas da raça Murrah sadias. As búfalas primíparas (G1) apresentaram maior atividade das enzimas gamaglutamiltransferase (GGT: 2.346U/L) e fosfatase alcalina (ALP: 181U/L) e maiores concentrações de fósforo (P: 56,6mg/dL), potássio (K: 32,0mg/dL) e α-lactoalbumina (458mg/dL). As fêmeas com duas a três lactações (G2) apresentaram maior CCS (70.700 células/mL) e maiores concentrações de proteína total (1,55g/dL), albumina (100mg/dL), magnésio (Mg: 8,80mg/dL), cloretos (Cl: 176mg/dL), ferro (Fe: 10,7µg/dL), sódio (Na: 178mMol/L) e lactoferrina (59,5mg/dL). As fêmeas com mais de três lactações (G3) apresentaram maiores concentrações de cálcio total (Ca: 41,8mg/dL), cálcio ionizado (Cai: 2,92mMol/L), imunoglobulina A (IgA: 1,32mg/dL), albumina sérica (99,1mg/dL), imunoglobulina G (IgG: 49,7mg/dL) e b-lactoglobulina (1.068mg/dL). Durante a lactação foi observado aumento da CCS, aumento das atividades das enzimas GGT e ALP, aumento das concentrações de proteína total, albumina, P, Mg, Cl, Na, lactoferrina, albumina sérica, IgG, α-lactoalbumina e redução das concentrações de Ca, Fe, Cai, K, IgA e b-lactoglobulina no soro lácteo das búfalas. Os resultados obtidos podem ser utilizados como referências para a espécie bubalina e auxiliar no diagnóstico e no prognóstico de doenças de ocorrência comum na fase de lactação.(AU)

To evaluate the biochemical profile and protein concentration of whey from milk samples of healthy Murrah primiparous and pluriparous buffaloes, 30 female buffaloes were analyzed during a complete lactation. The animals were divided into three groups: G1 = 10 primiparous buffaloes, G2 = 10 pluriparous buffaloes with 2-3 lactations and G3 = 10 pluriparous buffaloes with >3 lactations. The lactation period was divided into: early stage (I: 1-3 months of lactation), intermediate stage (T: 4-6 months of lactation) and final stage (F: 7-9 months of lactation). Before milk sampling, physical examination of the mammary gland, strip cup test and California Mastitis Test (CMT) were performed. After mammary quarters asepsis, 20mL of milk were collected monthly from each mammary quarter, during a complete lactation, in sterilized plastic bottles without preservative, in order to perform microbiological isolation, biochemical profile and protein electrophoresis in sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), and 30mL of milk from each mammary quarter were collect, in sterilized plastic bottles containing preservative bronopol to perform the somatic cell count (SCC). A total of 1,042 milk samples were collected from the experimental groups during lactation, of which 923 samples showed negative reaction to CMT and negative microbiological isolation and were selected to biochemical profile analysis and protein electrophoresis in SDS-PAGE. There were influence of parity order and stage of lactation in biochemical profile and protein concentration of healthy Murrah buffaloes' whey. Primiparous buffaloes (G1) showed higher gamma-glutamyltransferase (GGT: 2,346 U/L), alkaline phosphatase (ALP: 181 U/L), phosphorus (P; 56.6mg/dL), potassium (K; 32.0mg/dL) and α-lactalbumin (458mg/dL). Buffaloes with 2-3 lactations (G2) showed higher SCC (70,700 cells/mL) and higher concentrations of total protein (1.55g/dL), albumin (100mg/dL), magnesium (Mg; 8.80mg/dL), chlorides (Cl; 176mg/dL), iron (Fe; 10.7µg/dL), sodium (Na; 178mMol/L) and lactoferrin (59.5mg/dL). Bufalloes with >3 lactations (G3) showed higher concentrations of total calcium (Ca; 41.8mg/dL), ionized calcium (iCa; 2.92mMol/L), immunoglobulin A (IgA; 1.32mg/dL), serum albumin (99.1mg/dL), immunoglobulin G (IgG; 49.7mg/dL) and ß-lactoglobulin (1,068mg/dL). During lactation it was observed increase in SCC, GGT, ALP, total protein, albumin, P, Mg, Cl, Na, lactoferrin, serum albumin, IgG and α-lactalbumin, as well as decrease in concentrations of Ca, Fe, iCa, K, IgA and ß-lactoglobulin in buffaloes' whey. The results may be used as reference for buffaloes and to support diagnosis and prognosis of diseases common to lactation periods.(AU)

Latent tuberculosis in rheumatoid arthritis: evaluating cellular response and high-resolution computed tomography.

INTRODUCTION: The diagnosis of latent tuberculosis (LTB) in patients with rheumatoid arthritis (RA) has become important with the introduction of anti-tumor necrosis factor (anti-TNF-α) agents and the appearance of active tuberculosis cases in these patients. The tuberculin skin test (TST) has limited value in patients with RA. Tests based on the release of interferon-gamma (IFN-γ) are being studied, but their role has not been well established for this group of patients. OBJECTIVES: To compare the diagnosis of LTB in patients with RA by using cellular immune response to the TST and T.SPOT-TB. Additionally, findings of tomography studies compatible with LTB were used. METHODS: Clinical evaluation, TST, T.SPOT-TB and high-resolution computed tomography (HRCT) in a group of patients with RA at the University Hospital of the Federal University of Goiás. RESULTS: Response to the TST was lower in patients with RA (13.5%) compared to the predicted values of the general population. T.SPOT-TB identified a higher number of patients with LTB than the TST (36.8%). HRCT showed changes compatible with LTB in 52.9% of the patients, including 8 of the 11 patients with negative TST and T.SPOT-TB. CONCLUSIONS: The TST by itself is insufficient to diagnose LTB. A higher number of positive results were obtained with T.SPOT-TB when compared to the TST. Nevertheless, it was negative in a large percentage of patients with tomography findings consistent with LTB. HRCT is readily available in most large health-care centers and it could be incorporated into the diagnostic strategy for LTB in patients with RA.