Periparturient blood T-lymphocyte PD-1 and CTLA-4 expression as potential predictors of new intramammary infections in dairy cows during early lactation (short communication).

BACKGROUND: The periparturient period in dairy cows is marked by immunosuppression which increases the likelihood of infectious disorders, particularly also mastitis. An in-depth understanding of peripartum leukocyte biology is vital for the implementation of highly successful post-partum disease prevention measures. Immune checkpoint molecules, such as programmed death 1 (PD-1) and cytotoxic T-lymphocyte antigen 4 (CTLA-4), are critical inhibitory receptors expressed on immune cells, particularly T cells, that drive immunosuppressive signaling pathways. However, the potential role of immune checkpoint molecules expression in T-cells on udder health has never been explored. Thus, the association between the occurrence of new postpartum intramammary infections (IMIs) and the expression of programmed cell death protein 1 (PD-1) and cytotoxic T-lymphocyte-associated antigen-4 (CTLA-4) on blood T-cells during the peripartum period was investigated. RESULTS: In this study, the incidence of IMIs by any pathogen in early lactation was not associated with a higher expression of PD-1 and CTLA-4 in the periparturient period. However, the incidence of IMIs by major pathogens throughout the first month of lactation was significantly associated with higher expression of PD-1 at 14 days before calving (P = 0.03) and CTLA-4 at parturition (P = 0.03) by blood T-cells. Also, the expression of CTLA-4 at D0 (P = 0.012) by T-cells was associated with the occurrence of persistent IMIs during the first month of lactation. CONCLUSIONS: To our knowledge, this is the first report to investigate the expression of PD-1 and CTLA-4 by blood T-lymphocytes during the periparturient period in dairy cows and to explore their relationship with the incidence of new IMIs in the postpartum period. Thus, a comprehensive understanding of leukocyte biology during peripartum would appear to be a prerequisite for the identification of resilient dairy cows or targets innovative (immunological) non-antibiotic approaches in the transition period.

Estudo retrospectivo das afecções locomotoras em ruminantes atendidos na Faculdade de Medicina Veterinária e Zootecnia da Universidade de São Paulo entre 2000 e 2012 / Retrospective study of locomotor disorders in ruminants attended in the School of Veterinary Medicine and Animal Science, University of São Paulo from 2000 to 2012

Foi realizado o estudo retrospectivo das afecções do sistema locomotor de ruminantes atendidos no Serviço de Clínica de Bovinos e Pequenos Ruminantes da Faculdade de Medicina Veterinária e Zootecnia da Universidade de São Paulo (FMVZ-USP), no período de 2000 a 2012. Nesse período, foram atendidos 209 casos de animais com problemas lo- comotores, dos quais 62,7% localizados na região distal dos membros e 37,3% na região proximal. Na espécie bovina, com 121 (57,9%) casos atendidos, o comprometimento da região distal dos membros foi observado em 86 (71,07%) e da região proximal em 35 (28,93%) dos casos, respectivamente. A afecção mais frequente observada em bovinos foi a hiperplasia interdigital com 26,74% (23) dos casos atendidos. Entre as afecções na região proximal, houve maior ocor- rência de fraturas, com 48,6% (17) dos casos. Foram atendidos 88 (42,1%) pequenos ruminantes, apresentando lesões na região distal em 51,1% (45) dos casos e 48,9% (43) com lesões na região proximal. Nessas espécies, as lesões de maior ocorrência nas regiões distais e proximais foram, respectivamente, o foot-rot (60%) e as fraturas (77,4%). As afecções do sistema locomotor dos ruminantes foram pouco frequentes entre os animais atendidos no período estudado. Quanto ao prognóstico, foi bom nos animais acometidos com doenças podais, diferentemente das afecções proximais dos bovinos, principalmente fraturas, que apresentaram prognóstico mau.

This study aimed to perform a retrospective study of ruminants attended at the Clinic for Cattle and Small Ruminants (CBPR) of the Faculty of Veterinary Medicine (FMVZ), University of São Paulo (USP) with locomotor diseases from 2000 to 2012. During this period 209 cases were treated. It was found that cases located in the distal limb and in the proximal region were 62.7% and 33.7%, respectively. In bovines, 121 (57.9%) cases were treated, with 86 (71.07%) cases presented in the distal limb and 35 (28.93%) cases in the proximal region. The most common disease was interdigital hyperplasia with 23 (26.74%) cases treated. Fractures were the most frequent disease related to the proximal regioncorresponding to 17 (48.6%) occurrences. In small ruminants, 88 animals (42.1%) were treated with 45 (51.1%) cases in the distal region and the other 43 (48.9%) in the proximal region. In these species, the foot-rot (60%) and fractures (77.4%) was the most common diseases found in the distal and the proximal region, respectively. The disorders of the locomotor system of ruminants were uncommon in the CBPR. While the affected animals with claw diseases have a good prognosis, disorders affecting the upper limb in cattle, mainly fracture, have a poor prognosis.

Factors affecting immunoglobulin concentration in colostrum of healthy Holstein cows immediately after delivery / Fatores que afetam o nível de imunoglobulina no colostro de vacas holandesas sadias imediatamente após o parto

This study analyzed the influence of the number of milkings, number of births, and udder quarter in immunoglobulin (Ig) concentration in the colostrum of healthy Holstein cows. It was collected two samples of colostrum by manual milking, getting the first jets to completion of bacteriological examination and immunoglobulin levels by radial immunodiffusion test in agar gel. Positive samples for bacteriological examination were excluded from this investigation. Medians of immunoglobulin's G, A and M in the colostrum collected before the first and second milking were respectively 9,200 and 6,400mg/dL (p=0.0029); 400 and 200mg/dL (p=0.0018); 800 and 400mg/dL (p=0.0001). Median immunoglobulin concentration in animals that calved once, twice or three times or in cows that calved 4 to 6 times were 6,400; 6,400; 3,200 and 11,200mg/dL IgG; 100, 200, 100 and 800mg/dL IgA ; and 400, 400, 100 and 800mg/dL IgM, respectively. Concentrations of IgG, IgA and IgM were greater in animals that calved more than 4 times (p<0.05). Medians of IgG, IgA and IgM in the right fore quarter (RF), right hind quarter (RH), left fore quarter (LF) and left hind quarter (LH) were, respectively, 7,800; 6,400; 7,800 and 6,400mg/dL; 200, 200, 200 and 200mg/dL; and 400, 400, 400 and 400mg/dL. Ig concentrations in the colostrum of Holstein cows were influenced by the number of milkings after delivery and number of lactations. These variations may be considered risk factors to passive immunity transfer to newborn calves, predisposing them to diseases and causing economic losses to dairy production.

A pesquisa avaliou a influência do número de ordenhas, número de parições e quarto mamário na concentração de imunoglobulinas (Ig) do colostro de vacas hígidas da raça Holandesa. Foram colhidas duas amostras de colostro por ordenha manual, obtendo-se os primeiros jatos para a realização do exame bacteriológico e determinação dos níveis de imunoglobulinas pelo teste de imunodifusão em gel de ágar. As amostras positivas ao exame bacteriológico foram eliminadas desta investigação. Os valores medianos obtidos para a concentração de imunoglobulinas das classes G, A e M do colostro colhido antes da primeira e segunda ordenha foram, respectivamente de 9.200 e 6.400mg/dL (p=0,0029); 400 e 200mg/ dL (p=0,0018); 800 e 400mg/dL (p=0,0001), respectivamente. Os valores medianos da concentração de imunglobulinas, nos animais com apenas 1 parto, 2, 3 ou nas vacas com 4 a 6 partos foram de 6.400, 6.400, 3.200 e 11.200mg/ dL para a IgG; 100, 200, 100 e 800mg/dL para a IgA; e 400, 400, 100 e 800mg/dL para a IgM, respectivamente. As concentrações de IgG, IgA e IgM foram superiores nos animais com mais de 4 partos (p<0,05). Os valores medianos de IgG, IgA e IgM obtidos nos quartos mamários anterior direito (AD), posterior direito (PD), anterior esquerdo (AE) e posterior esquerdo (PE) foram respectivamente 7.800, 6.400, 7.800, 6.400mg/dL; 200, 200, 200, 200mg/dL; e 400, 400, 400 e 400mg/dL, não observando-se diferenças estatísticas (p>0,05) entre os quartos mamários. Os teores de Igs do colostro de vacas Holandesas sofrem influência do número de ordenhas pós-parto e número de lactações. Estas variações podem ser consideradas fatores de risco associados à falha na transferência de imunidade passiva em bezerros neonatos, predispondo-os às doenças e ocasionando perdas produtivas á pecuária.

Macrófagos lácteos de búfalas hígidas: avaliações da fagocitose, espraiamento e liberação de H2O2 / Milk macrophages from healthy female buffaloes: evaluation of phagocytosis test, spreading and hydrogen peroxide release assays

O presente estudo teve por objetivo avaliar funcionalmente os macrófagos lácteos “nonelicited” presentes por meio de testes de fagocitose, espraiamento e liberação de peróxido de hidrogênio. Foram colhidas 56 amostras de leite de 15 búfalas hígidas e mensuradas acontagem de células somáticas total e diferencial, a viabilidade celular,os testes de fagocitose, de espraiamento e a liberação de peróxido de hidrogênio. Dessas variáveis obteve-se respectivamente média de14.500 cél/mL de leite; com mediana de 4,33 de linfócitos e médias e desvios padrão de 50,77 + 18,28 de células da série monócito/macrófago e 32,13 + 19,27 de polimorfonucleares. A viabilidadedas células na suspensão foi 66,8 +15,8 e os índices de fagocitose e espraiamento foram 30,1 + 16,9 e 58,5 +13,3. Não houve diferençaentre a liberação de H2O2 espontânea e induzida por PMA. Concluiu-se que os macrófagos presentes no leite de búfalas hígidas e espraiaram significativamente, além de apresentarem correlação com outro marcador de ativação celular, no caso, a liberação de peróxido de hidrogênio; mais da metade dos macrófagos aderidos fagocitaram partículas de zymosan; os fagócitos mantêm sua capacidade de liberar peróxido de hidrogênio, espontaneamente ou não, em grau máximo,com uma significativa variação entre amostras.

The objective of the present study was to evaluate the functioning ofnonelicited dairy macrophages present in phagocytosis, spreading and hydrogen peroxide release tests. Fifty-six samples of milk were collected from 15 healthy buffaloes. Total and differential somatic cell counts, cell viability, and indexes of phagocytosis, spreading and hydrogen peroxide release were assessed. The following values were obtained: mean of 14,500 cells/mL of milk, with median equal to 4.33 of lymphocytes; mean and standard deviation equal to 50.77 + 18.28 of monocytes / macrophages and 32.13 + 19.27 polymorphouclear cells. Mean viability of cells in suspension was 66.8 +15.8; phagocytosis and spreading indexes were equal to 30.1 + 16.9 and58.5 +13.3, respectively. There was no difference between the spontaneous release of H2O2 and the one induced by PMA. It was concluded that nonelicited macrophages present in the milk of healthy buffaloes were significantly capable to spread and phagocyte. Phagocytes presented the ability to release hydrogen peroxide either spontaneously or not, in a maximum level and with a significant variation between samples.