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This research communication presents an automatic method for the counting of somatic cells in buffalo milk, which includes the application of a fuzzy clustering method and image processing techniques (somatic cell count with fuzzy clustering and image processing|, SCCFCI). Somatic cell count (SCC) in milk is the main biomarker for assessing milk quality and it is traditionally performed by exhaustive methods consisting of the visual observation of cells in milk smears through a microscope, which generates uncertainties associated with human interpretation. Unlike other similar works, the proposed method applies the Fuzzy C-Means (FCM) method as a preprocessing step in order to separate the images (objects) of the cells into clusters according to the color intensity. This contributes signficantly to the performance of the subsequent processing steps (thresholding, segmentation and recognition/identification). Two methods of thresholding were evaluated and the Watershed Transform was used for the identification and separation of nearby cells. A detailed statistical analysis of the results showed that the SCCFCI method is able to provide results which are consistent with those obtained by conventional counting. This method therefore represents a viable alternative for quality control in buffalo milk production.

This paper reports a case of abomasal bloat in a Holstein heifer, after an episode of the complete fracture of the left thoracic limb, which occurred on the farm. The animal, with 15 days of age, was donated to Veterinary Teaching Hospital (Hovet) of Faculty of Veterinary Medicine and Animal Science of University of Sao Paulo (FMVZ/USP) where it was operated, resulting in a complete fracture consolidation and no post-surgical complications. At 52 days of age, the heifer manifested acute apathy, anorexia, abdominal discomfort, and diarrhea. After physical and complementary (i.e., blood count and abdominal ultrasound) examinations and the assessment of nutritional management, the calf was diagnosed with abomasal bloat. Therefore, the following treatment was performed: correction of dehydration, antibiotic therapy, administration of a non-steroidal anti-inflammatory drug, and attempt of gas release through the esophageal tube. Moreover, dietary adjustments were made and consisted in interrupting the supply of concentrate, decreasing the volume of milk replacer, as well as supplying ruminal liquid, water, and green grass. Despite the acute and fatal characteristics seen in abomasal tympanism, the rapid identification of the initial clinical manifestations presented by the heifer allowed immediate therapeutic intervention, culminating in the survival of the animal and complete recovery at the end of the treatment.

RESUMO Objetivou-se avaliar o efeito dos anticorpos (ACs) maternos sobre resposta imune humoral induzida pela vacinação em bezerros Holandeses. Bezerros foram distribuídos aleatoriamente em quatro grupos: G1 - vacinados no D14 e D44 (n=6); G2 - vacinados no D90 e D120 (n=5); G3 - vacinados no D180 e D210 (n=8); controle: não vacinado (n=5). Utilizaram-se 5mL de vacina comercial (Cattle Master Gold FP5+L5® - Zoetis, Brasil), por via subcutânea. Foi realizada vírus neutralização (VN) no momento da vacinação, booster e 30 dias após a revacinação. Não foram observadas diferenças entre controle e G1 ou G2 para a frequência de soropositivos ou títulos de ACs contra os vírus respiratórios (P≥0,05). G3 apresentou maior produção de ACs em relação ao controle para BoHV-1 (P

This study aimed to examine the effects of selenium (Se) and vitamin E (vitE) supplementation on blood cell counts and blood metabolite concentrations in goats and their kids. Fifteen Saanen goats (average age 6 years of age; average initial body weight of 70 ± 10 kg) and 21 ½ Saanen × ½ Pardo Alpine crossbred goat kids (average body weight of 3.70 ± 0.64 kg) were used. Animals were distributed in a completely randomized design with five replicates per diet for mother goats and seven for goat kids and randomly assigned into three groups in the following diets: CON, control basal diet; Se, inclusion of 3.2 mg of Se/kg DM; SevitE, inclusion of 3.2 mg Se/kg DM and 1145 IU/day vitE/kg DM. Effects of time were observed on red blood cells, hemoglobin, hematocrit, mean corpuscular volume, and mean corpuscular hemoglobin in goats and goat kids. Effects of time were observed on differential counts of leucocytes, lymphocytes, and monocytes in goat kids. Interaction was observed for high-density lipoprotein and total protein in goats and for triglycerides, beta-hydroxybutyrate (BHBA), and gamma-glutamyltransferase (GGT) in goat kids. Effects of time were observed on low-density lipoprotein, triglycerides, glucose, lactate, BHBA, non-esterified fatty acids (NEFA), creatinine, aspartate-aminotransferase, and GGT in goats and all blood metabolites in goat kids. Selenium, vitE, or association in the evaluated levels are not sufficient to change blood cell counts when supplied in diets for goats or goat kids. However, the effect of time or interaction between time and diets change the blood metabolite concentrations in the animals.

The development and state of innate immune cell function during the first 90 days of life in dairy calves have not been fully described.This transversal study attempted to examine the changes that occur in circulating blood cells and the innate immune response in healthy calves from birth to 89 days of age.Healthy Holstein calves represent three windows of development, G1 from 1 to 7 days old (n = 26), G2 from 30 to 40 days old (n = 28), and G3 from 60 to 89 days old (n = 36) were sampled once each from a single herd. A few biomarkers of the general health and innate and inflammatory immune responses were measured.The youngest calves had the lowest red blood cell (RBC) counts, cell hemoglobin concentration means (CHCMs), red cell distribution widths (RDWs), and cell hemoglobin contents of mature red blood cells (CHm) compared with the other groups. They also had the lowest iron concentrations and highest intracellular myeloperoxidase indices. However, white blood cell (WBC) and lymphocyte concentrations gradually increased from G1 to G3. G2 calves had the lowest serum protein concentrations and highest number of innate immune cells compared with the other groups. Calves were able to mount phagocytic and ROS responses from birth.The physiologic responses of circulating blood cells and innate immune responses in dairy calves are shown according to age. Neonates had limitations in several RBC and WBC indices and immunologic responses that would likely impact overall vigor and health. Fortunately, these limitations resolve by 90 days of age.

Bovine respiratory disease (BRD) is a multifactorial disease which causes short- and long-term negative effects. Early detection is crucial for a prompt response to therapy, as well as to decrease mortality risk. Clinical scoring systems have been developed mostly in North America for screening calves at risk or suspected of having BRD, and these tools have also been applied in subtropical and tropical countries. However, it has been unknown whether these scoring systems had the same accuracy in tropical environmental conditions. Therefore, this study evaluated the accuracy of 4 different field techniques, as well as serum haptoglobin (HAP), to diagnose BRD in Holstein dairy calves in subtropical conditions. The tests used to diagnose BRD were thoracic ultrasound (TUS; positive if consolidation depth ≥1 cm), thoracic auscultation (AUSC; positive if crackles, wheezes, or silent areas were present), Wisconsin score (WISC; ≥2 categories with scores of ≥2), and California score (CALIF; positive if total score ≥5). Also, HAP was measured and classified as positive if ≥15 mg/dL. Heifers between 30 d of age and weaning (n = 482), residing on 17 commercial dairies in São Paulo state, were enrolled in this study. Bayesian latent class models were used with informative priors to evaluate the accuracy of TUS, AUSC, and HAP, and noninformative priors for the accuracy of WISC and CALIF. The percentage of calves positive for each test on each farm ranged from 0 to 56% for WISC, 11-51% for CALIF, 0-72% for TUS, 0-32% for AUSC, and 0-100% for HAP. The sensitivity (Se; 95% credible interval) and specificity (Sp) for WISC were 77.9% (64.8-90.2) and 81.9% (76.3-88.2). For CALIF, the Se was 67.1% (53.6-80.1) and Sp 79.1% (73.9-84.6). For TUS Se was 59.8% (46.5-73.1) and Sp was 84.8% (80.0-89.5), and for AUSC, Se was 58.8% (41.3-79.8) and Sp was 98.6% (95.7-99.9). The Se and Sp of HAP was 67.6% (55.3-78.8) and 46.7% (41.4-52.2), respectively. The performance of the scoring systems was similar to, or better than, the performance found in North American studies, despite the fact that calves were in a tropical environment.

Vaccination is a strategy to the prevention and control of reproductive diseases caused by bovine viral diarrhea virus (BVDV) and bovine herpesvirus type 1 (BoHV-1), however the various compositions of commercial vaccines should be evaluated for their ability to induce protection mediated by antibodies. The objective of this research was to evaluate the production of specific neutralizing Abs against BVDV-1 and 2, and BoHV-1 induced by commercial vaccines composed by different adjuvants. Holstein heifers were vaccinated and distributed in three experimental groups: Group I (G1) was vaccinated with a commercial vaccine containing inactivated BVDV-1, BVDV-2 and BoHV-1 diluted in alum hydroxide as adjuvant (n=9); Group II (G2) was vaccinated with an product containing inactivated strains of BVDV-1, BVDV-2, BoHV-1 and BoHV-5 diluted in oil emulsion as adjuvant (n=10); Group III (G3) was vaccinated with a commercial vaccine containing inactivated BVDV-1 and BVDV-2, besides live modified thermosensitive BoHV-1, diluted in Quil A, amphigen and cholesterol (n=10); A control, non-vaccinated group (n=6) was mock vaccinated with saline. Heifers received two subcutaneous doses of 5mL of each commercial vaccine on the right side of the neck, with 21 days interval. Humoral immune response was assessed by the virus neutralization test (VN) against BVDV-1 (NADL and Singer strains), BVDV-2 (SV253 strain) and BoHV-1 (Los Angeles strain) in serum samples collected on vaccination days zero (D0), 21 (D21) and 42 (D42; 21 days after boosting). Neutralizing Abs against BVDV-1 NADL was detected only in D42, regardless of the vaccine used. Similar geometric mean titers (GMT) for BVDV-1 NADL were observed between G1 (log2=5.1) and G3 (log2=5.1). The seroconversion rate (%) was higher in G1 (78%) when compared to G2 (10%) and G3 (40%). For BVDV-1 Singer, it was also possible to detect Abs production in G1 (log2=5.8, 100% seroconversion rate) and G3 (log2=3.5, seroconversion rate = 60%), only after the booster dose (D42). Neutralizing Abs to BVDV-2 (SV253) were detected only in G3, observing 90% seroconversion associated with high titers of Abs (log2=6.7) after the 2nd dose of vaccine (D42). Heifers from G1 and G3 responded to BoHV-1 after the first dose (D21): G1 (log2=2.5, seroconversion rate = 67%) and G3 (log2=0.7, seroconversion rate = 80%). In D42, a higher magnitude response was observed in the heifers from G3 (log2=6.1, 100%) compared with G1 (log2=4.3, 100%) and G2 (log2=2.7, 60%). Based on the data obtained, it can be concluded that the commercial vaccine contained aluminum hydroxide (G1) was most effective in the induction of antibodies against BVDV-1. On the other hand, this vaccine did not induce the production of neutralizing Abs against BVDV-2. Only the heifers from G3 (Quil A, amphigen and cholesterol) generated neutralizing Abs against BVDV-2. The animals that received commercial vaccine containing oil emulsion as adjuvant (G2) had a weak/undetectable response against BVDV-1 and BVDV-2. The best protective response against BoHV-1 was observed in heifers vaccinated with the live modified thermosensitive virus. RESUMO: A vacinação é utilizada como estratégia para a prevenção e controle das doenças reprodutivas, causadas pelos vírus da diarreia viral bovina (BVDV) e herpesvírus bovino tipo 1 (BoHV-1), entretanto, as diversas composições de vacinas comerciais devem ser avaliadas quanto a sua eficiência protetiva mediada por anticorpos (Acs). O objetivo desta pesquisa foi avaliar a produção Acs neutralizantes específicos para cepas de BVDV-1 e 2, e BoHV-1 induzida por vacinas comerciais contendo diferentes tipos de adjuvantes. Para tal, novilhas Holandesas foram vacinadas e distribuídas em três grupos experimentais: Grupo I (G1) foi vacinado com uma vacina comercial composta por cepas inativadas de BVDV-1, BVDV-2 e BoHV-1 diluídas em hidróxido de alumínio como adjuvante (n=9); Grupo II (G2) foi vacinado com produto contendo as cepas inativadas de BVDV-1, BVDV-2, BoHV-1 e BoHV-5 em uma emulsão oleosa como adjuvante (n=10); O Grupo III (G3) foi vacinado com uma vacina comercial contendo BVDV-1 e BVDV-2 inativado, além do BoHV-1 vivo modificado e termosensivel, diluídos em adjuvante contendo Quil A, Amphigem e colesterol (n=10); O Grupo Controle não vacinado (n=6) foi inoculado com solução salina. As novilhas receberam duas doses das respectivas vacinas ou solução salina (5mL), com intervalo de 21 dias, por via subcutânea, na tábua do pescoço do lado direito. A resposta imune humoral foi avaliada pelo teste de vírus neutralização (VN) contra o BVDV-1 (cepas NADL e Singer), BVDV-2 (cepa SV253) e BoHV-1 (cepa Los Angeles) em amostras de soro coletadas nos dias (D) de vacinação zero (D0), 21 dias após 1ª dose (D21)e 42 (D42; 21 dias após A 2ª dose). Os anticorpos neutralizantes contra o BVDV-1 NADL foram detectados apenas em D42, independentemente da vacina utilizada. Os títulos médios geométricos (GMT) de anticorpos foram semelhantes entre G1 (log2=5,1) e G3 (log2=5,1). A taxa de soroconversão foi maior no G1 (78%) quando comparado ao G2 (10%) e G3 (40%). Para o BVDV-1 Singer, somente após D42 foi observada a produção de Acs no G1 (log2=5,8; taxa de soroconversão de 100%) e G3 (log2=3,5; taxa de soroconversão = 60%). Os anticorpos contra BVDV-2 (SV253) foram detectados apenas nas novilhas do G3, observando-se taxa de soroconversão de 90% com altos títulos de anticorpos neutralizantes (log2=6,7) em D42. Novilhas G1 e G3 responderam ao BoHV-1 após a primeira dose (D21): G1 (log2=2,5; taxa de seroconversão = 67%) e G3 (log2=0,7; taxa de seroconversão = 80%). Em contrapartida, foi observada uma maior magnitude de resposta para as novilhas G3 (log2=6,1; 100%) em D42, em relação aos animais G1 (log2=4,3; 100%) e G2 (log2=2,7; 60%). Com base nos dados obtidos, foi possível concluir que a vacina composta por hidróxido de alumínio (G1) foi mais eficaz na produção de anticorpos contra o BVDV-1, em contrapartida esse produto não induziu anticorpos contra o BVDV-2. Apenas as novilhas do G3 (Quil A, amphigen e colesterol) geraram Acs neutralizantes contra o BVDV-2. Os animais que receberam a vacina em emulsão oleosa (G2) como adjuvante apresentaram uma resposta fraca/indetectável contra o BVDV-1 e BVDV-2. A melhor resposta protetiva contra o BoHV-1 foi observada nas novilhas vacinadas com a vacina viva modificada termosensível.

This objective of this study was to evaluate the use of tulathromycin on the timing of appearance and number of four indicator organisms representing the gastrointestinal microbial community, the incidence of diarrhea and a measure of the systemic inflammatory profile in Holstein heifers. Twenty-six Holstein heifer calves were distributed between receiving (ATB+) or not receiving (ATB-) tulathromycin at a dose of 2.5 mg/kg by 12 h of age. Samples from the calves were collected at six times during the neonatal period. Stool samples were used to determine the dry matter content and quantitative analysis of specific indicator bacterial populations. Samples of whole blood and serum were collected to determine the total number of neutrophils, the number of CD62L+ neutrophils, quantity of haptoglobin, and to allow for ex vivo measurement of reactive oxygen species. A higher frequency of diarrhea was detected in the ATB+ calves (84.6%) than ATB- (53.8%) on days 13–15 (P = 0.084). ATB- calves had a greater number of Bifidobacterium in stool on day 3–5 (P = 0.002), and on days 7–9 (P = 0.018). The ATB+ calves tended to have a higher number of Escherichia coli in stool on days 20–23 and days 27–30 (P = 0.052 and P = 0.072). Both the total number of neutrophils (P = 0.013) and the capacity for ROS production was higher in ATB- (P = 0.038) than ATB+ calves at all points tested. ATB+ calves had higher levels of haptoglobin (P = 0.032) on days 13–15. Administration of tulathromycin appeared to negatively impact the establishment of a normal microbiome and to modulate the development of innate immune function.

Background : Physiological and immunological adaptations occurred in peripartum period may be responsible for susceptibility to intramammary infections during colostrogenesis. Increase in blood flow and hormones cause physiological edema by accumulation of lymphatic fluid in interstitial space. Specific composition of mammary secretion increases the susceptibility to mastitis. So, the study of physical, cellular and microbiological changes in the mammary gland in this period are fundamental for understanding, diagnosing and adopting prophylactic measures for bovine mastitis. This study aimed to characterize physiological and pathological changes that occur in the mammary gland during transition period. Materials, Methods & Results : Thirteen cows, in second to fourth lactation were followed from 3 weeks pre-calving to 3 weeks post-calving. Cows were submitted weekly to physical examination of mammary gland, strip cup test, California Mastitis Test (CMT), Microscopic Somatic Cell Count (MSCC), and bacteriological examination of mammary secretion. Mammary quarters were divided into two groups: BAC - : samples that did not present bacteriological growth throughout transition period; BAC + : samples that presented bacteriological growth in at least two isolates of the same bacterial genus. Regarding physical examination of mammary gland, the parameters localized or generalized alteration in volume, asymmetry, reddish color, reduced skin elasticity, increased temperature, and swollen consistency presented higher frequencies in the BAC - group. On the other hand, infection in mammary quarters of cows from BAC + group promoted a decrease in frequencies of increased volume, decreased elasticity, and elevated temperature in the mammary gland, in addition to a higher positivity to CMT and higher cellularity. Overall rate of intramammary infection was 25.82%, with non-aureus Staphylococci (NAS) being the predominant agent (69.2%). Maximum peak of bacterial isolates occurred immediately after calving (33.3%). Differences were observed between groups ( P = 0.021) for CMT, showing a higher frequency of positive tests in BAC + group (36.5%) compared to BAC - group (18%). MSCC varied between weeks ( P = 0.001), groups ( P = 0.001), and between the interaction of the two variables ( P = 0.019). Highest median value of MSCC was observed at calving (1.5 × 10 6 cells/mL), being different ( P = 0.001) from the +1, +2, and +3 weeks and the lowest median value of MSCC (0.08×10 6 cells/mL) was observed in +3 compared to previous weeks ( P < 0.05). Discussion : NAS was predominantly isolated in mammary secretions of cows during peripartum, it is commonly found in the skin and canal of the teat and are associated with subclinical mastitis. Peak of mammary infection was observed on calving, probably by the cortisol peak resulting in a stress leucogram and decrease in the functional activity of immune cells. Changes observed in the physical examination of mammary gland of cows from BAC - group are compatible with physiological edema caused by circulatory changes that occurs with the proximity of calving, increasing blood flow necessary for production of colostrum. Alterations in the mammary quarters of cows from BAC + group may be associated with functional loss of mammary parenchyma or persistent infectious process, characterizing clinical findings of pathological changes during transition period. Mammary quarters of the BAC + group presented higher percentages of positive reactions to CMT in weeks +1 and +2, and higher MSCC values and are associated with infection of the mammary gland.

Background: Newborn calves are born immunosuppressed, hypogammaglobulinemic, immunologically immature, and therefore more vulnerable to many infectious diseases. During pregnancy, the fetal-placental environment is regulated by Th2-type cytokines that neutralize Th1 responses, an important factor for immune defense against viral agents. The ingestion and absorption of colostral immunoglobulins enhance the immunity of the neonate. However, the presence of maternal antibodies might negatively affect the success of parental vaccination in the first two months of life. This study aimed to evaluate the effecacy of parenteral vaccination in newborn calves with high titers of maternal antibodies against respiratory viruses. Materials, Methods & Results: Twenty-eight Holstein calves were allocated to the vaccinated group (VAC, n = 18) or an unvaccinated control group (NVAC, n = 10). The initial vaccination with 5 mL of a commercial vaccine occurred around the 14th day of life (D14) and the booster at D35. Respiratory and diarrhea symptoms were evaluated at D12, D14, D16, D20, D31, D36, D45, D53, and D60. Blood samples were taken for leukogram, haptoglobin, and seroneutralization of BVDV, BoHV-1, BRSV, and BPI3V, at the time of vaccination at D14 (T1), at booster (D35, T2), and 21 days after the booster (D56, T3). Despite the increased prevalence of BRD during the period of the study, no calves from either group exhibited respiratory disease at D12 or D14. In subsequent assessments, the frequency of BRD increased over time in the VAC group until it reached a maximum prevalence of 38.9% (7/18) at D31. In the NVAC group, the maximum prevalence observed was 40% at D45 and D60. A comparison of the frequencies for BRD cases showed a statistical trend at D36 (P = 0.07), with a higher prevalence for the NVAC group (30%) in relation to the VAC group (5.6%). For the NVAC group, a greater number of total leukocytes was observed at T3 (P = 0.038) and granulocytes (trend) at T2 (P = 0.066). Time analysis demonstrated a decrease in haptoglobin concentration in both groups (NVAC, P = 0.005 and VAC, P = 0.006), with a reduction in the values at T1 and T3 (NVAC = 0.005 and VAC = 0.024). Antibody titers were similar between groups for BVDV, BoHV-1, and BRSV. Higher titers for BPI3V were observed for the VAC group at D56 (P = 0.045). Discussion: The early-onset BRD was present in both groups between 30 and 60 days of age, based on the higher prevalence observed. These data reinforce the importance of early immunization programmes before infection. Factors such as management practices and facilities may have also contributed to the higher prevalence of BRD. The increased number of leukocytes and neutrophils at T2 (D35) and T3 (D56) in the NVAC group points to a stronger inflammatory response to various types of potential challenges. The inflammatory leukocyte profile explains the higher haptoglobin values observed in the NVAC group at T2 (D35). The similarity of titers of antibodies against BVDV, BoHV-1, and BRSV between groups may indicate that vaccinating the calves at 14 days of age did not induce a humoral immune response to these viruses, likely due to interference from the maternal antibodies. Early vaccination prevented a drop in specific viral antibodies and promoted partial protection for vaccinated calves around 14 days of age, with a decrease in the intensity of the inflammatory response at the peak of the disease and a higher concentration of antibodies against BPI3V after the booster. Fil: Gomes, Viviani. Universidade de Sao Paulo; Brasil Fil: Gonzalez, Diego Daniel. Instituto Nacional de Tecnologia Agropecuaria. Centro de Investigacion En Ciencias Veterinarias y Agronomicas. Instituto de Virologia E Innovaciones Tecnologicas. - Consejo Nacional de Investigaciones Cientificas y Tecnicas. Oficina de Coordinacion Administrativa Pque. Centenario. Instituto de Virologia E Innovaciones Tecnologicas.; Argentina Fil: Medici Madureira, Karina. Universidade Federal da Bahia; Brasil Fil: Mozgovoj, Marina Valeria. Instituto Nacional de Tecnologia Agropecuaria. Centro de Investigacion En Ciencias Veterinarias y Agronomicas. Instituto de Virologia E Innovaciones Tecnologicas. - Consejo Nacional de Investigaciones Cientificas y Tecnicas. Oficina de Coordinacion Administrativa Pque. Centenario. Instituto de Virologia E Innovaciones Tecnologicas.; Argentina Fil: Costa Baccili, Camila. Universidade de Sao Paulo; Brasil Fil: Toledo Silva, Bruno. Universidade de Sao Paulo; Brasil Fil: Silva Ramos, Jean. Universidade de Sao Paulo; Brasil Fil: Nascimento Silva, Karen. Universidade de Sao Paulo; Brasil Fil: Maristella Pituco, Edviges. Instituto Biológico; Brasil

A Brucelose Bovina é uma doença de importância econômica e de caráter zoonótico, considerada de notificação obrigatória, causada pela Brucella abortus, caracterizada principalmente por abortamento em terço final de gestação e afecções reprodutivas. O presente artigo relata o trabalho conjunto da Universidade, da Defesa Agropecuária e da Vigilância Epidemiológica em um caso de brucelose ocorrido na Clínica de Bovinos e Pequenos Ruminantes da Universidade de São Paulo. Um bovino com afecção locomotora foi diagnosticado com brucelose ao ser testado juntamente com outro animal da propriedade que apresentava afecção reprodutiva. A Defesa Agropecuária foi acionada e o animal foi encaminhado para abate sanitário. Foi realizada ação conjunta para monitoramento da brucelose na propriedade e notificação ao Serviço de Saúde do Município. Seis animais da propriedade e três pessoas apresentaram resultado positivo, ressaltando o potencial zoonótico da doença e a importância do médico veterinário no controle da mesma nos animais e no homem.

This research evaluated the effect of milking on the physical exam of the mammary gland (MG) and milk examination in Saanen goats. Six properties were selected, four farms were used after evaluation of criteria selection and distributed in two groups according to the milking method: manual or mechanical. The physical examination of MG was performed using the techniques of inspection and palpation, in addition to the examination of the milk by strip cup, California Mastitis Test, somatic cell count, and bacteriological exam. It was possible to detect an association between the milking method with the most of physical exam of the MG: inspection of the MG (p = 0.001), inspection of the teat (p = 0.002), palpation of the MG (p = 0.054) and palpation of the teat (p = 0.036). The machine milking had an association with the reduction of the volume of MG (OR = 6.00), reduction of the teat size (OR = 16.19), and firm consistency of mammary parenchyma (OR = 2.39). The use of machine milking represented a less risk for an increase of the volume of the MG volume (OR = 0.288) and the presence of cisternitis (OR = 0.490). The association between the SCC and the type of milking was also detected (p = 0.002). Machine milking was associated with changesin the physical exam of MG and milk examination compatible with traumatic lesions. This information indicates inappropriate use of milking equipment in the dairy goat’s properties.

The objective was to evaluate the effects of injectable trace minerals (ITM) concurrent with modified-live virus (MLV) vaccination on protection from bovine viral diarrhea virus (BVDV) infection in dairy calves. In a previous study (Palomares et al., 2016), thirty dairy calves received two doses of a MLV vaccine subcutaneously (SC), concurrently with ITM (n = 15) or saline (n = 15), SC. Five months later, 20 of these calves received ITM (G1, n = 10) or saline (G2, n = 10) according to their previous groups and were challenged intranasally with BVDV2. Five unvaccinated calves were also challenged with BVDV2 (G3). Blood samples were collected on days 0 (BVDV challenge), 3, 5, 6, 7, 8, 9, 11, 14, 18, 21, 32 and 61 for leukocyte count, virus isolation and BVDV serum neutralizing antibodies (SNA). Mild-moderate clinical signs were observed in G3 after BVDV challenge. Group 1 showed lower sum health score and nasal score on d5 and fecal score on d8 compared to G2. Rectal temperature and leukocyte counts were not different between G1 and G2. In contrast, G3 calves had significant leukopenia and lymphopenia from d3 to d7 (P .05) and higher rectal temperatures on d6 to d8, compared to values on d0 (P .05). All unvaccinated calves became viremic, while viremia was not detected in G1 or G2. Average daily gain was not different between vaccinated groups, however, only G1 calves had significantly greater (P = .04) ADG compared to non-vaccinated calves during the first 14 days post challenge. Vaccinated calves treated or not with ITM were protected from BVDV2 infection five months post-vaccination.

The aim of this research was to evaluate the influence of maternal cells from colostrum on the development and function of the innate immune response in Holstein calves. Calves were divided into 2 groups: COL + (n = 10) received fresh colostrum; and COL − (n = 10) which received frozen colostrum containing no viable cells. The calves were assessed before colostrum intake (D0), 48 h of age (D2), and weekly from D7 up to D28. Blood samples were collected for analysis of the distribution of leukocytes, cellular phenotype and in vitro granulocyte function. COL + calves tended to have a high number of neutrophils on D7 (p = 0.073). COL − calves took up significantly more Escherichia coli (measured as MFI) on D7 (p = 0.034). Endogenous production of radicals (as percentage of cells) tended to be higher in COL − calves on D14 (p = 0.061). The intensity of endogenous reactive oxygen species (ROS) produced by granulocytes tended to be higher in COL + calves on D21 (p = 0.094). Overall, ROS production (percent of cells, and MFI) induced by Staphylococcus aureus and Escherichia coli were higher in COL + calves than COL − calves. It was our observation that COL + calves developed an innate immune response more quickly and efficiently after natural exposure to pathogens after birth. In contrast, COL − calves mounted an innate response more slowly that yielded a persistent inflammatory response after natural exposure to these bacteria agents. This research provides evidence of an advantage to the calf of receiving fresh colostrum on the development and function of the innate immune system., Highlights • Calves fed whole fresh colostrum (COL +) developed a fast and efficient innate immune response. • Calves fed frozen colostrum (COL −) colostrum had a slow and persistent inflammatory response after natural challenge. • COL + calves demonstrates advantages in relation to innate immune response against pathogens causing diarrhea.

The objective of this research was to evaluate the influence of cells from colostrum on the health of neonate calves. Animals were distributed in 2 groups: COL + (n = 9) which received fresh colostrum from their own damns; and COL − (n = 10) which received frozen colostrums from donors. Heifers were assessed before colostrum intake – D0; D2; D7; D14; D21 and D28. Heifers were monitored by clinical examination, hematological profile and serum iron. COL − had a higher diarrhea intensity score (typically 3) on D7. Moreover, a single case each of bronchopneumonia and navel inflammation were observed in COL − calves. COL − had fewer red blood cells (RBC) (6.5 ± 0.8 × 106/μL) and less hemoglobin (Hgb) (8.3 ± 1.4 g/dL) than COL + (RBC = 7.2 ± 0.8 × 106/μL; Hgb = 9.6 ± 1.3 g/dL) at D14 (P ≤ 0.05). COL − had more anemia on D21 (P = 0.03) and on D28 (P = 0.02). Iron was lower in COL − (5.6 ± 2.7 μM/L) than COL + (10.7 ± 6.2 μM/L) (P = 0.03) on D7. Lymphocytes was lower in COL − than COL + on D7 (3.8 ± 1.0 × 103/μL COL + and 5.4 ± 2.2 × 103/μL COL −, P = 0.02). COL − calves had more anemia and lower serum iron concomitant with diarrhea on D7. The number of leukocytes was relatively consistent in the COL + calves, while COL − calves showed an increasing number of of lymphocytes starting on D7., Highlights • Calves fed with frozen colostrum (COL −) had high intensity of disease. • COL − had anemia and low serum iron. • COL − had higher number of lymphocytes (D7) due to natural exposure to pathogens.

After vaccination, vaccine components must activate the immune response, but the ideal vaccine should not result in undesirable effects in cattle. The aim of this study was to evaluate the inflammatory and humoral responses and adverse reactions induced by three adjuvanted commercial vaccines against bovine viral diarrhea virus (BVDV) and bovine herpesvirus 1 (BHV-1). Holstein heifers (n = 35) were divided into four groups by adjuvant compounds: Vaccine A (Alum; n = 9), Vaccine B (Oil-in-water; n = 10), Vaccine C (Amphigen/Quil A cholesterol and dimethyl-dioctadecyl ammonium (DDA) bromide (QAD; n = 10), and Control (n = 6). Heifers were assessed at 0 h, 6, 24, 48, 72 and 168 h post-vaccination; serology was evaluated at first dose (D0), booster (D21) and D42. Heifers vaccinated with Vaccine B (p = 0.0001) and C (p = 0.0001) had a more intense local reaction, while there was a higher rectal temperature detected in heifers vaccinated with Vaccine C (p = 0.020). There was greater systemic reaction observed for heifers vaccinated with Vaccines B and C at 48 h (p = 0.002) after a second dose. Clinical pathology parameters [white blood count (WBC) (p = 0.001), neutrophils (p = 0.0001) and haptoglobin concentrations (p = 0.0001)] were higher in animals vaccinated with Vaccine C. Neutralizing Abs against BVDV type 1 strains, NADL and Singer, were detected in animals vaccinated with Vaccines A or C at D42, while BVDV-2 antibodies were detected only in animals vaccinated with Vaccine C. A BHV-1 antibody was detected in all three vaccine groups (Vaccines A, B or C) at day 42 (21 days post booster vaccination). The findings of this research were based on three different commercial laboratory formulations and also according to the conditions which the study was conducted. In this context, vaccine containing mineral oil or Amphigen/QAD presented greater local reactivity and induced a significant systemic inflammatory response. Vaccinated heifers with Alum and Amphigen/QAD commercial vaccines enhanced humoral immune response against BVDV and BHV-1.

O objetivo deste estudo foi a comparação das metodo-logias mais comuns empregadas em diferentes fazendas para a avaliação da qualidade do colostro e identificação de falha na transferência de imunidade passiva (FTIP) em bezerras Holandesas. Sessenta e oito bezerras Holandesas recém-nascidas foram distribuídas em dois grupos: G1 - provenientes de propriedade com manejo de colostro adequado; G2 - oriundas de propriedade com falhas no manejo do colostro. Foram colhidas amostras de colostro fornecido para cada bezerra de cada grupo (250 ml) para a avaliação de IgG, sólidos totais, PT e albumina. Amostras sanguíneas foram obtidas às 48 horas de vida para a determinação de PT e de globulina e para a estimativa indireta de IgG por meio dos testes de turvação pelo sulfato de zinco, precipitação pelo sulfito de sódio e coagulação do glutaraldeído. Os teores de IgG observados no colostro pelo colostrômetro (G1: 98,63 ±1 5,58 mg/ml; G2: 85,66 ± 29,81 mg/ml; p = 0,09), índice Brix (G1: 27,61 ± 2,96%; G2: 22,40 ± 4,58%; p = 0,00), PT (G1: 10,42 ± 2,29g/dL; G2: 9,04 ±2 ,29g/dL; p = 0,06) e albumina (G1: 1,25 ± 0,37g/dL; G2: 0,91 ± 0,38g/dL; p = 0,00) foram superiores no G1 quando comparados ao G2. A determinação de PT no soro das bezerras foi realizada por um analisador bioquímico automático, revelando taxas de FTIP (< 5,2 g/dL) somente no G2 (46,67%). Os resultados dos testes qualitativos (turvação pelo sulfato de zinco, precipitação pelo sulfito de sódio e coagulação do glutaraldeído) foram diferentes dos resultados de PT (p = 0,00) e não se mostraram eficientes para a detecção da FTIP. Esta pesquisa demonstrou que além do uso do colostrômetro para a avaliação da qualidade do colostro, a utilização do refratrômetro Brix e níveis de PT do colostro devem ser consideradas. A identificação de FTIP no rebanho pode ser realizada seguramente por meio dos níveis séricos de PT e globulina.

RESUMO O objetivo deste trabalho foi avaliar, de forma longitudinal, o perfil sérico proteico de 13 vacas Holandesas durante o período de transição. Amostras de sangue (n=78) foram coletadas semanalmente, da segunda semana pré-parto (M-2) até a terceira semana pós-parto (M3), para determinação do perfil sérico proteico, por meio de teste bioquímico (proteínas séricas totais - PT) e eletroforese em gel de poliacrilamida SDS-PAGE, para as outras proteínas analisadas. Os valores de PT diminuíram de forma gradativa (P=0,000) de M-2 (6,4g/dL) a M0 (6,2g/dL), aumentando nos momentos subsequentes (M3=7,3g/dL). As concentrações da IgG de cadeia pesada (M-2=919,4; M-1=1074,5mg/dL) e de cadeia leve (M-2=393,9; M-1=466,7mg/dL) foram menores no pré-parto em relação ao pós-parto (M1=1283,3; M2=1374,2 e M3=1630,3 mg/dL para IgG pesada e M1=463,4; M2=573,7; M3=651,8mg/dL para IgG leve). Para a IgA, houve diminuição nos valores (P=0,001), de M-2 (51,9mg/dL) a M1 (34,0mg/dL), e aumento em M2 (45,4g/dL) e M3 (62,6g/dL). Os valores de haptoglobina (Hp) e ceruloplasmina (Cp) aumentaram (P=0,000) de M-2 (Hp=16,6mg/dL; Cp=8,6mg/dL) a M3 (Hp=60,9mg/dL; Cp=127,1mg/dL). A albumina apresentou ligeiras variações durante o período de transição (P=0,000), enquanto a transferrina sérica (P=0,101) e a glicoproteína ácida (P=0,105) foram estáveis. O escore de condição corporal (ECC) também foi analisado durante o período de transição, verificando-se diferença (P=0,003) entre M-2 (ECC=4,0) e M1 (ECC=3,0). Foi relatada ainda a ocorrência de distocias (4/13), retenção de placenta (1/13) e hipocalcemia (1/13) no dia da parição (M0) e infecções uterinas (5/13) e cetose (1/13) ocorridas no pós-parto. Concluiu-se, portanto, que houve aumento nas concentrações séricas de Hp e Cp e diminuição nos valores de imunoglobulina e transferrina em vacas Holandesas no período de transição, relacionados às doenças ocorridas nesse período e ao elevado ECC, promovendo modificações metabólicas e imunossupressão. ABSTRACT The aim of this study was to evaluate longitudinally the serum protein profile of 13 Holstein cows during the transition period. Blood samples (n=78) were taken weekly, from the second week before parturition (M-2) to the third week after parturition (M3) for determination of serum protein profile by biochemical tests (total serum protein - PT) and SDS-PAGE electrophoresis, for the other proteins analyzed. PT values decreased gradually (P = 0.000) from M-2 (6.4g/dL) to M0 (6.2g/dL), increasing in subsequent moments (M3 = 7.3g/dL). The concentrations of the heavy chain (M-2=919.4; M-1=1074.5mg/dL) and the light chain of IgG (M-2=393.9; M-1=466.7mg/dL) were lower in pre-calving compared to post calving (M1= 1,283.3; M2=1,374.2 and M3=1,630.3 mg/dL for the heavy chain, and M1=463.4; M2=573.7 and M3=651,8 mg/dL for the light chain of IgG). For IgA there was a decrease in the values (P=0.001), from M-2 (51.9mg/dL) to M1 (34.0mg/dL), and increase in M2 (45,4g/dL) and M3 (62,6g/dL). The values of haptoglobin (Hp) and ceruloplasmin (Cp) increased (P=0,000) from M-2 (Hp=16.6mg/dL; Cp=8.6 mg/dL) to M3 (Hp=60.9mg/dL; Cp=127.1mg/dl). Albumin showed slight variations during the transition period (P=0.000), while the serum transferrin (P=0.101) and acid glycoprotein (P=0.105) were stable. Body condition score (BCS) was also analyzed during the transition period, checking the difference (P=0.003) between M-2 (ECC= 4.0) and M1 (ECC=3.0). It was also reported the occurrence of dystocia (4/13), retained placenta (1/13) and hypocalcemia (1/13) on the day of calving (M0) and uterine infections (5/13) and ketosis (1/13) occurring in the post calving. In conclusion, there was an increase in serum Hp and Cp, and a decrease in the immunoglobulins and transferrin amounts in Holstein cows during the transition period, related to diseases occurring during this period and the high BCS, promoting metabolic changes and immunosuppression.

RESUMO Esta pesquisa avaliou a TIP e a dinâmica de anticorpos (ACs) específicos em bezerros naturalmente expostos aos agentes causadores da doença respiratória bovina (DRB). Foram selecionados 19 bezerros Holandeses alimentados com colostro proveniente de doadoras vacinadas para DRB. Amostras de soro foram obtidas antes e após a ingestão do colostro (48h) para a soroneutralização (SN). Os valores médios (log2) detectados após colostragem foram de 11,5±1,6 (BVDV), 8,8±1,3 (BoHV-1), 5,5±1,6 (BRSV) e 8,4±1,5 (BPIV-3). Cinco bezerros foram criados do nascimento aos 240 dias de vida, observando-se decréscimo nos títulos de ACs para BVDV, BoHV-1 e BPIV-3 ao longo do tempo (P≤0,001). As taxas de infecções detectadas entre o D14 e o D240 foram de 40% (2/5), 20% (1/5), 80% (4/5), e 60% (3/5), respectivamente, para BVDV, BoHV-1, BRSV e BPIV-3. A maioria dos bezerros manifestou broncopneumonia após as infecções virais. Os bezerros apresentaram ACs para todas as viroses às 48 horas de vida, porém os títulos adquiridos para o BRSV foram baixos. A susceptibilidade para as infecções variou de acordo com os níveis e a duração dos títulos de ACs maternos. ABSTRACT This research evaluated the PIT and the dynamics of specific antibody (Ab) for calves naturally exposed to the viral agents involved in Bovine Respiratory Disease (BRD). Nineteen Holstein calves fed colostrum from vaccinated donors for DRB. Serum samples were obtained before and after colostrum intake (48h) for serum neutralization (SN). Mean values (log2) detected after colostrum feeding were 11.5±1.6 (BVDV), 8.8 ±1.3 (BoHV-1) 5.5±1.6 (BRSV) and 8.4±1.5 (BPIV-3). Five calves were raised from birth to 240 days of life and presented a decrease in Ab titers for BVDV, BoHV-1 and BPIV-3 over time (P≤ 0.001). Infection rates from D14 to D240 were of 40% (2/5), 20% (1/5), 80% (4/5) and 60% (3/5), respectively for BVDV, BoHV-1, BRSV and BPIV-3. Most of the calves presented bronchopneumonia after seroconversion to the virus. Calves presented Ab for all viruses at 48 hours of life, however BRSV Ab titer were low. Levels and persistence of maternal antibody titers determined the susceptibility to viral infections.

RESUMO: O periparto caracteriza-se por mudanças imunes e fisiológicas na glândula mamária das fêmeas bovinas, tornando-as mais susceptiveis às mastites. Este trabalho teve o objetivo de avaliar a fagocitose da bactéria Escherichia coli pelos fagócitos do sangue e do colostro de vacas hígidas da raça Holandesa. Amostras provenientes de 22 fêmeas, entre novilhas e vacas, foram obtidas durante a primeira ordenha pós-parto, para avaliação da fagocitose, em lamínula de vidro, após estímulo com E. coli. A proporção de fagócitos espraiados do sangue foi maior quando as células foram estimuladas com bactérias (p=0,003), no entanto a taxa de fagocitose foi baixa e semelhante à observada quando as células não foram estimuladas (p=0,299). No colostro, foram constatados maiores índices de fagocitose (p=0,001) e espraiamento (p=0,000) quando os fagócitos foram estimulados com E. coli. Comparando-se os resultados obtidos para o sangue e o colostro, verificou-se, no sangue, maior proporção de fagócitos não espraiados e que não apresentavam bactérias no seu interior, estimulados (p=0,000) ou não (p=0,002) com bactéria. Houve maior taxa de espraiamento nos fagócitos do colostro quando estimulados com E. coli (p=0,002). Não se viu diferença nos percentuais de fagocitose entre as amostras de sangue e colostro incubadas (p=0,478) ou não (p=0,071) com bactérias. Os resultados permitiram concluir que E. coli influenciou no espraiamento dos fagócitos do sangue e do colostro de fêmeas bovinas da raça Holandesa no período do pós-parto imediato e auxiliarão na compreensão dos mecanismos imunológicos ocorridos nesse período, considerado o de maior susceptibilidade das fêmeas bovinas às infecções. ABSTRACT: The peripartum is characterized by immune and physiological changes in the mammary glands of cows, making them more susceptive to mastitis. This work aimed to evaluate the phagocytosis of Escherichia coli by phagocytes from blood and colostrum from healthy Holstein cows. Samples from 22 female, heifers and cows, were obtained during the first postpartum milking for evaluation of phagocytosis in glass cover slip, after stimulation with E. coli. The proportion of spreading blood phagocytes was higher when cells were stimulated with bacteria (p=0.003). However, the phagocytosis rate was low and similar to that observed when blood cells were unstimulated (p=0.299). Colostrum showed higher rates of spreading (p=0.000) and phagocytosis (p=0.001) when phagocytes were stimulated with E. coli. Comparing the results obtained for blood and colostrum, a higher proportion of not spreading phagocytes without bacteria inside, when stimulated (p=0.000) or not (p=0.002) with bacteria, was observed in blood. There was a higher spreading rate in colostrum when phagocytes were stimulated with E. coli (p=0.002). There was no difference in the percentage of phagocytosis in blood or colostrum samples incubated (p=0.478) or not (p=0.071) with bacteria. The results showed that E. coli influenced the activity of blood and colostrum phagocytes from Holstein cows in the immediate postpartum period, and will assist in the understanding of the immunological mechanisms that occurs in this period, considered as the most susceptible to infections in cows.

RESUMO: Este trabalho avaliou a influência da infecção bacteriana da glândula mamária (GM) sobre a transferência de imunidade passiva (TIP) em bezerros recém-nascidos. Vacas holandesas (n=13) foram observadas no momento da parição e ordenhadas para a obtenção de forma asséptica das amostras de colostro (n=52) para os testes microbiológicos. Os recém-nascidos receberam 6 litros de colostro de uma ordenha nas primeiras 12 horas de vida, proveniente de suas respectivas mães. Amostras de sangue foram colhidas antes (D0) e após (D2) o manejo do colostro. A TIP foi avaliada por meio de testes bioquímicos, eletroforese e leucograma. Os bezerros foram distribuídos conforme a ausência (IB-) ou presença (IB+) de infecção mamária em pelo menos uma GM de suas respectivas mães. Todas as amostras de colostro (n=52) foram negativas ao cultivo fúngico. Das 13 fêmeas, 8 (61%) apresentaram crescimento bacteriano em ≥1 quartos mamários. Considerando-se os quartos mamários, foi obtido isolamento bacteriano em 21,15% (11/52), observando-se predomínio de espécies bacterianas do grupo Staphylococcus coagulase negativa. Não foram encontradas diferenças entre os parâmetros de acordo com os grupos experimentais. Em relação aos momentos, foi possível verificar aumento nos valores de proteína total, globulinas, atividade sérica da gama glutamiltransferase e frações eletroforéticas beta e gamaglobulina após a ingestão do colostro materno. A mastite subclínica não influencia a transferência de imunidade passiva em bezerros recém-nascidos da raça holandesa, avaliados por teste bioquímicos, eletroforese e leucograma. ABSTRACT: This study evaluated the influence of intramammary infection under the passive immunity transfer in newborn calves. Holstein cows (n=13) were monitored at delivery, and milked to obtain sterile colostrum samples (n=52) for microbiological examination. The newborn calves received 6 liters of colostrum from their dams up to 12 hours after birth. Blood samples were harvested before (D0) and after (D2) colostrum feeding. The passive immune transfer was evaluated by biochemical tests, electrophoresis and leukogram. Calves were distributed according to absence (IB-) or presence (IB+) of mammary gland infection in one or more quarters. All colostrum samples (n=52) were negative for fungal culture. Eight dams (8; 61%) had bacterial growth in one or more mammary quarter. In relation to mammary gland unit, bacterial growth was observed in 21.15% (11/52) of the colostrum samples, with predominance of coagulase negative staphylococcal infections. Statistical differences were not found among variables according to the groups. According to the experimental moments, the evaluated parameters (total protein, albumin, globulins, GGT and beta and alpha electrophoretic) increased after colostrum intake. Subclinical mastitis did not influence the passive immune transfer in Holstein calves evaluated by biochemical tests, electrophoresis or leukogram.

The objective of this study was to evaluate the influence of viable cells from colostrum on immune development in dairy heifer calves during the first 28 days of life. The animals were distributed between 2 groups: COL+ (n=9) receiving fresh whole colostrum from their own damns; and COL- (n=10) receiving pooled frozen colostrum, containing no viable cells, from a pool of donor cows. These calves were assessed before colostrum intake (D0), 48 hours of age (D2), and weekly from D7 to D28. The development of immunity was evaluated by assessment of the phenotype of blood leukocyte subsets, and induced cytokine production after 72 hours of stimulation in culture with concanavalin A (ConA), killed Staphylococcus aureus (S.aureus) and killed Escherichia coli (E. coli) by peripheral blood mononuclear cell (PBMC). The clinical history of these calves was marked by a high frequency of diarrhea in both groups. However, COL- had greater diarrhea intensity scores (fecal score~3 of 4), and rectal temperature on D7 than COL+ calves. Moreover, bronchopneumonia (n=1) and navel inflammation were observed only in COL- calves. COL- had a lower concentration of serum iron, and a higher absolute number of lymphocytes on D7 than COL+. COL- also had a higher percentage of anemic calves than the COL+ calves on D21 and D28. In general, the percent of cells within each subset of leukocytes was similar between the groups over the experiment, except on week 1 when COL- calves had a higher percentage of lymphocytes expressing CD45RO+ (P=0.07). A steady increase in CD45RO+ and concomitant decline in CD45RO- leukocytes was observed over the course of the study, indicating the development of immune memory. The proportion of CD14MHCII+ leukocytes increased with age (P≤0.05). The median background cytokine production by PBMC that were not stimulated was below the level of detection of the assays used for both groups. The PBMC from COL+ calves stimulated with ConA secreted a larger quantity of IL-17 week 2 (COL+=2060.0pg/mL and COL-=0.0pg/mL, P=0.00). PBMC from COL+ calves stimulated with killed S. aureus whole cell antigen (P=0.05) and killed E. coli whole cell antigen (P=0.05) also secreted higher levels of IL17 than COL- calves at week 4. Clear production of IL17 was observed in PBML from COL+ calves at week 2, but the difference was not statistical different between groups. In conclusion, calves fed fresh and frozen colostrum showed no difference in cells subset profile overall. The increased percentage of leukocytes expressing the memory CD45RO+ or CD14MHCII+ over the course of the experiment indicated a maturation of the adaptive immune response after natural exposure to pathogens in the environment of the calf. The enhanced IL17 secretion by COL+ calves indicated that viable maternal cells modulated T-cell Th17 production that was primed by bacterial antigens. This mechanism could be responsible for quick and efficient activation of neutrophils for bacterial clearance. The differences in cytokine production observed between groups may help to explain the different clinical pictures observed for calves COL+ and COL- calves.

O objetivo deste trabalho é apresentar um método de segmentação e contagem de células somáticas de leite bovino para imagens obtidas diretamente do microscópio óptico por meio de uma proposta de limiarização automática. Para fins de verificar eficiência da técnica, o método de limiarização proposto foi comparado com dois métodos de limiarização reconhecidos pela literatura. A correlação obtida entre a contagem manual de referência e os métodos de limiarização testados foi de 0,98, 0,71 e 0,12, respectivamente. O método proposto apresentou correlação aceitável demonstrando eficiência deste algoritmo para limiarização e contagem de células somáticas e outras aplicações similares.

During 24 months 96 marsupials and 64 rodents were captured, inspected and set free in the same space of the Pedra Branca State Park, Rio de Janeiro. From them, 105 ticks from 10 species in two families were manually collected. The dominant tick species on the rodents was Amblyomma longirostre and on the marsupials was Ixodes loricatus. There was a significant correlation of the temperature in relation to intensity of parasitism for ticks, demonstrating that the higher the temperature, greater is the number of ticks.

Bovine colostrum contains immune factors, like leukocytes and cytokines that are absorbed by the intes tinal mucosa of the neonate, however, not much is known about the function of some of these components in th e active immune response of calves. The aim of this s tudy was to determine the influence of colostrum in take and development of the innate immune response of calves from birth to two months of age, by evaluating the proportion of neutrophils CH138 + and monocytes CD14 + , using flow citometry and measurement of proinflammatory cytokines IL-1 beta, IL-6, TNF-alpha and IFN-gamma, using comercial ELISA kits. According to the obtained results we can determine that colos trum intake had influenced the IFN-gamma concentration and the monocytes CD14 + and neutrophils CH138 + proportions in the circulation of calves and that these components seem to play a key role in the development of immune response of these animals, providing protection against pathogens, enhancing the phagocytic activity and an tigen presentation by monocytes.

Changes in the composition of colostrum in the peripartum period focused on the neonate’s immunity can minimize the response of the mammary gland before natural exposure to mastitis pathogens. This research evaluated the oxidative activity of mononuclear and polymorphonuclear phagocytes of bovine colostrum. Bacteriologically negative (n = 171) bovine colostrum samples of first and second milking postpartum were evaluated. The oxidative activity was assessed by the production of hydrogen peroxide (H2O2) released either spontaneously or upon stimulation by enterotoxigenic Escherichia coli (ETEC). For this, three treatments were used: control (C) – only cells; non-opsonized (NO) - cells and ETEC non-opsonized; and opsonized (O) – cells and ETEC opsonized. Median values of concentrations (nmol/106) of H2O2 produced by cells obtained from the first and second milking treatments C, NO and O were 149.1 and 39.9 (P = 0.0007); 125.5 and 49.1 (P = 0.0007); 102.4 and 54.4 (P = 0.008), respectively. No differences were observed between the treatments at each milking. The amount of H2O2 produced at first milking exceeded the values found thereafter. Furthermore, the presence of bacteria did not determine the increased amount of H2O2 released by phagocytes. Thus, we concluded that the mammary gland’s events during the peripartum period have an effect on the proportion and activity of phagocytes, which can cause injuries to the breast parenchyma by the large amount of free radicals produced. The high frequency of bovine mastitis during this period indicates the need for studies of the immunity of the mammary gland and research of susceptibility factors for bacterial infections.

A fim de estabelecer os valores hematologicos e bioquimicos de ovinos da raca Dorper e avaliar a influencia do sexo e idade nestas determinacoes, 72 animais foram distribuidos em tres grupos experimentais de acordo com a faixa etaria, sendo o Grupo 1 (G1) composto por animais com ate 12 meses de idade (n=28), o Grupo 2 (G2) composto por animais de 13 a 36 meses de idade (n=37) e o Grupo 3 (G3) por animais de 37 a 60 meses de idade (n=7). Os animais foram distribuidos ainda em outros dois grupos exprimentais, de acordo com o sexo, sendo o Grupo 4 (G4) composto por femeas (n=57) e o grupo 5 (G5) por machos (n=15). Avaliando-se os resultados obtidos para os exames hematologicos e bioquimicos verificou-se que os valores medios de hemacias (13,4 ± 2,5x106/mL), hemoglobina (12,6 ± 1,3 g/dL), volume globular (37,5 ± 3,7%), leucocitos totais (7.116,0 ± 1.967,0/mL) e neutrofilos segmentados (4.168,0 ± 1.501,0/mL) foram maiores nos animais do G1, assim como a ureia, GGT e FA (52,0 ± 17,0 mg/dL, 93,0 ± 19,0 UI / L e 257,0 ± 125,0 UI / L, respectivamente). Comparando-se os sexos, verificou-se que os valores de leucocitos totais (7.576,0 ± 2.082,0/mL), neutrofilos segmentados (4.296,0 ± 1.510,0/mL) e linfocitos (2.428,0 ± 1.084,0/mL) foram maiores nos machos (G5). Os valores de creatinina (1,3 ± 0,4 mg/dL), proteinas totais sericas (6,3 ± 0.9 g/dL) e globulinas (3,7 ± 1.0 g/dL) foram maiores nas femeas (G4). A analise estatistica dos resultados encontrados permitiu determinar os valores de referencia para os constituintes hematologicos e bioquimicos de ovinos da raca Dorper, alem de concluir que ocorreram variacoes nos referidos parâmetros, influenciados pelo sexo e por diferentes faixas etarias.

In order to establish hematological and biochemical Dorper sheep values and evaluate the influence of sex and age in these determinations, 72 animals were assigned to three experimental groups according to age: Group 1 (G1) consisting of animals up to 12 months of age (n=28), Group 2 (G2) composed of animals from 13 to 36 months (n=37) and Group 3 (G3) for animals from 37 to 60 months (n=7). The animals were still divided in another two groups according to gender: Group 4 (G4) – females (n=57) and Group 5 (G5) – males (n=15). Evaluating the results obtained for hematological and biochemical examinations it was found that the mean values of RBC (13.4 ± 2.5x106/mL), hemoglobin (12.6 ± 1.3 g/dL), packed cell volume (37.5 ± 3.7%), total leukocytes (7116.0 ± 1967.0/mL) and segmented neutrophils (4168.0 ± 1501.0/mL) were higher in the animals of G1, as well as urea, GGT and ALP (52.0 ± 17.0 mg/dL, 93.0 ± 19.0 IU/L, 257.0 ± 125.0 IU/L, respectively). Comparing the gender, it was found that the values of total leukocytes (7576.0 ± 2082.0/mL), segmented neutrophils (4296.0 ± 1510.0/mL) and lymphocytes (2428.0 ± 1084.0/mL) were greater in males (G5). The values of creatinine (1.3 ± 0.4 mg/dL), serum total protein (6.3 ± 0.9 g/dL) and globulins (3.7 ± 1.0 g/dL) were higher in females (G4). Statistical analysis of the results allowed to determine reference values for hematological and biochemical constituents of Dorper sheep breed, and concluded that there were variations in these parameters influenced by gender and different age groups. A fim de estabelecer os valores hematológicos e bioquímicos de ovinos da raça Dorper e avaliar a influência do sexo e idade nestas determinações, 72 animais foram distribuídos em três grupos experimentais de acordo com a faixa etária, sendo o Grupo 1 (G1) composto por animais com até 12 meses de idade (n=28), o Grupo 2 (G2) composto por animais de 13 a 36 meses de idade (n=37) e o Grupo 3 (G3) por animais de 37 a 60 meses de idade (n=7). Os animais foram distribuídos ainda em outros dois grupos exprimentais, de acordo com o sexo, sendo o Grupo 4 (G4) composto por fêmeas (n=57) e o grupo 5 (G5) por machos (n=15). Avaliando-se os resultados obtidos para os exames hematológicos e bioquímicos verificou-se que os valores médios de hemácias (13,4 ± 2,5x106/mL), hemoglobina (12,6 ± 1,3 g/dL), volume globular (37,5 ± 3,7%), leucócitos totais (7.116,0 ± 1.967,0/mL) e neutrófilos segmentados (4.168,0 ± 1.501,0/mL) foram maiores nos animais do G1, assim como a ureia, GGT e FA (52,0 ± 17,0 mg/dL, 93,0 ± 19,0 UI / L e 257,0 ± 125,0 UI / L, respectivamente). Comparando-se os sexos, verificou-se que os valores de leucócitos totais (7.576,0 ± 2.082,0/mL), neutrófilos segmentados (4.296,0 ± 1.510,0/mL) e linfócitos (2.428,0 ± 1.084,0/mL) foram maiores nos machos (G5). Os valores de creatinina (1,3 ± 0,4 mg/dL), proteínas totais séricas (6,3 ± 0.9 g/dL) e globulinas (3,7 ± 1.0 g/dL) foram maiores nas fêmeas (G4). A análise estatística dos resultados encontrados permitiu determinar os valores de referência para os constituintes hematológicos e bioquímicos de ovinos da raça Dorper, além de concluir que ocorreram variações nos referidos parâmetros, influenciados pelo sexo e por diferentes faixas etárias.

The somatic cell count (SCC) is a screening test for the evaluation of intramammary infection; however, changes in mammary secretion during colostrogenesis can promote a physiological increase in the SCC, potentially reducing its reliability in the diagnosis of mastitis. Therefore, the aim of this study was to evaluate bovine colostrum SCC as an indicative parameter of breast infection in Holstein periparturient cows. A total of 80 samples were harvested from the first milking colostrum of 20 cows and were subjected to manual SCC and bacteriological examination. Bacterial growth was present in 36.62% of the crops; coagulase-negative Staphylococcus (CNS) was the predominant microorganism (76.92%). The median SCC in infected cows (1.8 × 106 cells/mL) was significantly higher than in uninfected cows (0.9 × 106 cells/mL) (p = 0.0451). The sensitivity (100–15%), specificity (100–2.2%), and false positive (100–2.2%) of the SCC decreased gradually when thresholds of 0.2–10.0 × 106 cells/mL were adopted. In contrast, the proportion of false negatives (0–84.6%) revealed an opposite trend. The threshold of greatest concordance between SCC and bacteriological examination was 10.0 × 106 cells/mL; however, the sensitivity rates (15.4%), specificity (2.2%), and false positive (2.2%) were very low. Based on these results, we conclude that SCCs increased prior to the infectious processes of the mammary gland, particularly in the CNS group. However, physiological changes caused by colostrogenesis resulted in poor concordance between the SCC and bacteriological examination of the colostrum. A contagem de células somáticas (CCS) é um teste de triagem para avaliação de infecção intramamária, porém, as alterações na composição do colostro observadas durante a colostrogênese podem promover o aumento fisiológico da CCS e inviabilizar o uso desta prova para diagnóstico da mastite. Assim, o objetivo desta pesquisa foi avaliar a CCS do colostro como parâmetro indicativo de infecção mamária em vacas Holandesas periparturientes. Para tanto, foram colhidas 80 amostras de colostro de primeira ordenha, provenientes de 20 vacas, para a CCS manual e exame bacteriológico (EB). Observou-se crescimento bacteriano em 36,62% dos cultivos, com predomínio de Staphylococcus coagulase-negativa (SCN) (76,92%). O valor mediano da CCS de vacas infectadas (1,8 x106 células/mL) foi maior do que o obtido para as vacas sadias (0,9 x106 células/mL) (P=0,0451). A sensibilidade (100 a 15%), especificidade (100 a 2,2%) e falso-positivo (100 a 2,2%) diminuíram gradativamente quando os limiares de 0,2 a 10,0 x106 células/mL foram adotados. Em contrapartida, a proporção de falso-negativo (0 a 84,6%) apresentou perfil inverso. O limiar de maior concordância entre a CCS e EB foi de 10,0 x106 células/mL, porém os índices de sensibilidade (15,4%), especificidade (2,2%) e falso-positivo (2,2%) foram muito baixos. Com base nos resultados encontrados, conclui-se que a CCS apresentou elevação diante dos processos infecciosos da glândula mamária (GM), causados especialmente por bactérias do grupo SCN. No entanto, as alterações fisiológicas decorrentes da colostrogênese resultaram em baixa concordância entre a CCS e o exame bacteriológico do colostro.

To establish reference values and to assess the influence of age on the leukograms of healthy Holstein calves, blood samples were obtained from 300 animals. These samples were distributed equally (n=20) among 15 experimental groups according to age: birth to 8 hours, 9 to 16 hours, 17 to 24 hours, 2 days, 3 days, 4 days, 5 days, 6 to 7 days, 8 to 9 days, 10 to 11 days, 12 to 13 days, 14 to 15 days, 16 to 20 days, 21 to 25 days and 26 to 30 days of age. The maximum numbers of leukocytes (9,305.0/mL), segmented neutrophils (6,551.2/mL) and total neutrophils (6,678.3/mL) were noted within the first 8 hours of life, while band neutrophils peaked in number (133.3/mL) between 9 and 16 hours after birth. Meanwhile, the maximum total lymphocyte (4,992.1/µL) and typical lymphocyte (4,686.1/µL) counts occurred between 21 and 25 days, whereas atypical lymphocytes (388.5/µL) reached their maximum number between 26 and 30 days, demonstrating an inversion of the neutrophil:lymphocyte ratio over time. Thus, the influence of age on the leukocyte count of the evaluated calves was verified. The release of endogenous corticosteroids during labor or at birth may contribute to this variation in leukograms with age. Com a finalidade de estabelecer os valores de referência e de avaliar a influência do fator etário sobre o leucograma de bezerras sadias, da raça Holandesa, utilizaram-se amostras de sangue de 300 animais, distribuídos com igual número (n=20) por 15 grupos experimentais, de acordo com a idade: do nascimento até 8 horas de vida; de 9 até 16 horas; de 17 até 24 horas; 2 dias de idade; 3 dias; 4 dias; 5 dias; 6 a 7 dias; 8 a 9 dias; 10 a 11 dias; 12 a 13 dias; 14 a 15 dias; de 16 a 20 dias; de 2l a 25 dias; e de 26 a 30 dias de vida. Puderam-se observar valores máximos nas primeiras oito horas de vida para leucócitos (9305,0/µL), neutrófilos segmentados (6551,2/µL) e neutrófilos totais (6678,3/µL); entre 9 a 16 horas pós-nascimento para os neutrófilos bastonetes (133,3/µL). Para os linfócitos totais (4992,1/µL) e os linfócitos típicos (4686,1/µL) os valores máximos ocorreram entre 21 e 25 dias, e para os linfócitos atípicos (388,5/µL), entre 26 e 30 dias, demonstrando-se uma inversão na relação neutrófilo: linfócito, com o decorrer dos dias de vida. Constatou-se a da influência do fator etário no leucograma dos bezerros avaliados, no entanto acredita-se que a liberação de corticoides endógenos no momento do parto/nascimento, contribuíram para esta variação.

The purpose of the present trial was to compare the percentages of necrotic and apoptotic polymorphonuclear leukocytes (PMNL) in goat milk with low and high somatic cell count (SCC). Twenty eight milk samples were collected from 20 lactating goats, determined to be negative in bacteriological examination, and divided in three groups, according to their SCC: samples with SCC lower than 500 × 10 3 cells/mL; between 500 and 1500 × 10 3 cells/mL; and higher than 1500 × 10 3 cells/mL. SCC was performed in an automatic somatic cell counter. Apoptosis and necrosis were quantified using dual-color flow cytometry with fluorescein labeled annexin-V and propidium iodide (PI). Results of the present study showed a significant positive correlation between the percentage of the viable PMNL and milk SCC ( r = 0.495, P = 0.008), as well as a significant negative correlation between apoptotic PMNL and milk SCC ( r = −0.486, P = 0.009). Results also pointed out lower PMNL viability rates due to higher apoptosis rates in milk samples with SCC lower than 5 × 10 5 cells/mL.

Background: Physiological events occur in the transition period marked by negative energetic balance, where the energetic demand is higher due to fetal nutrition and lactogenesis and intensified by the decrease of the dry matter intake in the pre-partum period. The adaptation of cows is dependent of homeostatic and homeorhetic mechanisms regulated by catecholamine, cortisol, GH, IGF-I, insulin and glucagon but the priorization of homeorhetic mechanisms could result in metabolic diseases. Considering the scarce longitudinal studies about the Brazilian conditions the aim of this project was to evaluate the influence of transition period on metabolic indicators, cortisol and IGF-I in Holstein cows.Materials, Methods & Results: Thirteen cows, from 2nd to 4th lactation were evaluated weakly from week -2 to week +3 in relation to calving to determine the body condition score (BCS), metabolic and endocrine profile. The BCS decreased between week -2 (BCS = 4.0) and +1 (BCS = 3.0), followed by and slight increase on weeks +2 and +3 (P = 0.000). The most of metabolic indicators and hormones have changed during the transition period (P ≤ 0.05), especially around calving. It was possible to detect the peak of cortisol, glucose, non-esterified fatty acids (NEFA), ß-hydroxibutyrate (BHB), and aspartate aminotransferase (AST) on delivery and week +1. On the other hand, total calcium, triglycerides (TG) and cholesterol (CHOL) decreased was observed around parturition. Insulin like growth factor type I (IGF-I) showed marked reduction between pre and postpartum, and the lowest value was observed in the week +1. TP and GLOB had the lowest value on calving and globulin has a gradual increase from delivery to week +3.Discussion: Cows included in this experiment had dystocia (4/13, 30.8%), retention of placenta (1/13, 7.7%) and clinical hypocalcemia (1/13, 7.7%). One of the animals with episodes of dystocia was also aggravated by ketosis in week +1 and +2 (1/13, 7.7%). Uterine infections were observed in five animals (5/13, 38.4%). Of this total of animals with uterine infection (5/13), three had already expressed earlier disturbances: two cows had dystocia and one cow with retention of placenta. The weight loss observed between pre and postpartum may be due to an increase of energy demand for fetal maintenance and the concomitant initiation of lactation, in conjunction with an approximately 30% reduction in dry matter intake during the transitional period. The hyperglycemia concomitant with the peak of cortisol at delivery may be explaining by the activation of the somatotropic axis, due to the stress of calving. The increase of NEFA, BHB and AST around calving associated with decreased of TG and CHOL could be associated with homeorhetic mechanisms, especially lipolysis and gluconeogenesis. The IGF-I decrease in postpartum may be by the decreased expression of receptors for growth hormone (GH) in the liver, and consequently reduced synthesis and/or IGF-I hepatic clearance, which reduce of the negative feedback of IGF-I on GH secretion. The total calcium concentrations were below the threshold (8.50 mg/dL) established by Goff [10] at parturition, week +1 and +2 and probably is due to the secretion of 20-30 g of calcium per day during the colostrum synthesis. Total protein and albumin have a slight variation, except for the peak observed on week +3 due to uterine infection.Keywords: body condition score, non-esterified fatty acids, ß-hydroxibutyrate, total calcium, hormones.

O eritrograma é uma avaliação que representa um auxílio ao clínico no diagnóstico de afecções que acometem os animais domésticos. No entanto, estudos hematológicos em bubalinos são escassos, havendo poucas informações na literatura a respeito de valores de referência dos constituintes sanguíneos desta espécie. Assim, este trabalho teve como objetivo a análise do eritrograma de búfalos de diferentes faixas etárias da raça Murrah, machos e fêmeas. Os animais em estudo foram distribuídos em quatro grupos experimentais, de acordo com as idades: Grupo 1: animais com idade entre o nascimento e 3 meses (n=15); Grupo 2: animais com idade entre 4 e 6 meses (n=50); Grupo 3: animais com idade entre 7 e 12 meses (n=50); e Grupo 4: animais com idade superior a 12 meses (n=50). Como resultados das análises foram encontrados os seguintes valores médios, no Grupo 1: 7,9x106 hemácias/mL de sangue (He); 13,0 g/dL de hemoglobina (Hb); hematócrito (Ht) de 38,9%; Volume Corpuscular Médio (VCM) de 49,0 fl; Concentração Hemoglobínica Corpuscular Média (CHCM) de 33,6 %, e Hemoglobina Corpuscular Média (HCM) de 16,4 pg; no Grupo 2: He: 7,1 x10(6)/mL; Hb: 12,5 g/dL; Ht: 36,8%; VCM: 52,4 fl; CHCM: 33,9%, e HCM: 17,8 pg; no Grupo 3: He: 7,9 x10(6)/mL; Hb: 12,0 g/dL; Ht: 33,8%; VCM: 43,1 fl; CHCM: 35,4%, e HCM: 15,34 pg; e no Grupo 4: He: 6,7 x10(6)/mL; Hb: 11,7 g/dL; Ht: 34,4%; VCM: 53,4 fl; CHCM: 34,4%. e HCM: 17,4 pg. A análise estatística dos resultados encontrados para as diferentes faixas etárias permitiu concluir que ocorreram variações determinadas pela evolução da idade, caracterizadas por diminuição do número médio de hemácias (He), da concentração de hemoglobina (Hb) e do valor do hematócrito (Ht). Os índices hematimétricos apresentaram variações significativas no G3.

Dissertação (mestrado) - Universidade Federal de Santa Catarina, Centro de Ciências Biológicas, Programa de Pós-Graduação em Ecologia, Florianópolis, 2015. Áreas recifais marginais estão sendo consideradas em todo mundo como áreas prioritárias para conservação, já que essas áreas possuem um grande potencial evolutivo das espécies. Tem sido vista também uma ampliação de distribuição de espécies tropicais nessas áreas, em função das mudanças ambientais. Recentemente, foi encontrado o único banco de corais rolados (banco de coralitos) do Atlântico Subtropical na Reserva Biológica Marinha do Arvoredo, no litoral de Santa Catarina ? Brasil. Os recifes rochosos de Santa Catarina são considerados o limite sul de distribuição de várias espécies tropicais de peixes, ascídias, esponjas e corais, entre outros. O banco é formado por colônias livres roladas do coral Madracis decactis, que estruturam um ambiente diferente dos costões rochosos adjacentes. O presente estudo avalia o recrutamento dos organismos sésseis associados ao banco de coralitos e sua composição. O recrutamento dos organismos foi avaliado com experimento ?in situ? ao longo de 12 meses com placas de recrutamento em inclinações, distâncias do costão e tempos de submersão das placas de recrutamento distintos. Conjuntos de placas foram retirados com 3, 6 e 12 meses de submersão e analisados para investigar a composição e variação no recrutamento. Observamos que há padrão na distribuição das espécies principalmente em relação ao tempo de submersão das placas, com as maiores variações na composição da comunidade entre as placas com 3 e 6 meses submersão, se estabilizando a partir das placas com 6 meses de submersão, sem grandes alterações quando comparadas as placas com 12 meses de submersão. O fator inclinação e a distância do costão também apresentaram valores significativos com relação a variação na taxa de recrutamento. Encontramos 14 grupos taxonômicose/ou funcionais durante o experimento sendo que desses grupos, cirripedia, biofilme e macroalga coriácea obtiveram maior correlação com o tempo de submersão em que as placas foram expostas. Os grupos ascidiacea, bryozoa e macroalga crostosa obtiveram maior correlação com relação a inclinação, onde a macroalga crostosa obteve as maiores porcentagens de cobertura nas placas com inclinação horizontal sendo que os outros grupos citados na inclinação vertical. Os grupos cirripedia e hydrozoa apresentaram maior correlação com o fator distância do costão, sendo encontrados em maior número nas placas mais afastadas do costão. A composição da comunidade se altera em função das condições ambientais e das relações interespecíficas, sendo independente da composição inicial. Os resultados vão de encontro com a teoria de sucessão, onde o tempo é o principal fator explicando as mudanças na composição da comunidade. Não foi observado nenhum recruta do coral M. decactis nas placas de recrutamento.. O estudo revela um ambiente vulnerável já que a espécie principal estruturadora do ambiente está em situação frágil no ponto de vista ecológico, com uma população pequena, de lento crescimento e aparentemente com pouca ou nenhuma contribuição da reprodução sexuada. Novas medidas de manejo devem ser adotadas para que haja efetividade na conservação desse ambiente único. Abstract : Marginal reef areas are now considered priority areas for conservation around the world because of the great evolutionary potential of associated species. Tropical species distributions have recently expanded in these areas as a result of environmental change. Recently, the only Atlantic subtropical coralith bed was found in the Arvoredo Marine Biological Reserve on the coast of Santa Catarina, Brazil. The rocky reefs of Santa Catarina are considered the southern boundary for distribution of various species of tropical fish, sea squirts, sponges, and corals, among others. The bank is formed by free colonies of the coral Madracis decactis, which structure an environment quite different from the adjacent rocky shores. This study evaluates the recruitment of sessile organisms associated with the coralith bed, and species composition. Recruitment was assessed via "in situ" experiments in which recruitment plates were placed on different slopes, distances from shore, and submersion times; results were evaluated over 12 months. Plate sets were removed at 3, 6 and 12 months post-submergence for investigation of variation in community composition and recruitment. We found the greatest differences in plate community composition between 3 and 6 months post-immersion; this difference stabilized at 6 months of submersion without further major differences. Different slope angles and distances from shore produced significant variation in recruitment rate. We found 14 taxonomic and/or functional groups during the experiment and of these groups, cirripeds, biofilm and coriaceous macroalgae showed the greatest correlations with plate submergence time. Ascidiacea, bryozoa and crustose algae were the groups most correlated with slope, and crustose macroalgae had the highest percentages of coverage on the plates with horizontal slopes, with the other groups were found in greater abundances on vertical slopes. Cirripeds and hydrozoa showed strong correlation with distance from the shore, with greater numbers on plates further from the rocky shore. Changes in community composition depended on environmental conditions and interspecific relationships, independent of initial composition. These results are in accordance with the succession theory,in which time is the primary explanatory factor for changes in community composition. New M. decactis recruits were not found on experimental plates. The study reveals a vulnerable environment as the main structuring species seems to be in an ecologic fragile situation, with a slow-growing, small population and poor or lack of sexual reproduction. . New management measures must be applied in order to promote effective conservation of this unique environment.

A two-step methodology for background removal of bovine milk images is proposed.A well-tuned k-means algorithm removes all sort of debris from background.A new thresholding removes the debris-free and statistically varying background.The background-removed cells under Watershed transform are robust-segmented.High accuracy somatic cell counting algorithm for bovine milk quality control. Mastitis is an infectious disease associated with the increased number of somatic cells in cow's milk, and it is one of the most relevant cause of economic losses in dairy farming industries. In this paper, we propose a method capable of determine, with 99.7% accuracy, the number of these cells in microscope slide images. This level of accuracy is achieved by changing the image original RGB format to Lab color space and applying k-means clustering algorithm to remove debris and other background features. A new gray level thresholding is proposed, and the remaining bound cells are separated in the final segmentation step applying Watershed transform. Many microscope slide images with debris, contrast, and hue variation were used to validate the experimental results. Comparison between the proposed method and manual counting indicates that this new approach is a robust and promising solution to be incorporated in a future automated somatic cell counting system using optical microscopy.

The objective of the present trial was to evaluate the effect of the stage of lactation on somatic cell counts (SCC) in milk produced by healthy goats. Forty goats that presented no clinical signs of udder inflammatory process, and whose milk was negative in bacteriological examination were used. All animals were kept in the same management system and received the same diet. They were submitted to monthly collections of milk for 8 months. Milk was analyzed by means of the California Mastitis Test (CMT), automatic SCC and bacteriological examination. The following values were recorded from the first to the eighth month of lactation: 2.56, 4.42, 3.45, 8.52, 5.96, 5.84, 6.41 and 6.51 × 10 5 somatic cells/mL of milk, respectively. When the first trimester and the period between the fifth and the eighth month of lactation were compared, there was an increase in cell counts. Percentage of 1+, 2+ and 3+ CMT reactions increased from the sixth month of lactation on. Results showed that there was an increase in the number of somatic cells in milk as lactation went by, which was demonstrated by both counting methods. © 2005 Elsevier B.V. All rights reserved.

Background: Infections are caused by Bovine Viral Diarrhea Virus and still continue to be a worldwide plague in cattle industry. It is responsible for sudden death syndromes in adult cattle with high mortality rates, abortions, acute gastrointestinal and respiratory diseases. The BVDV infection occurs in early pregnancy (40-142 days), in immunosuppressed females or cows results in 100% of persistently infected (PI) calves that are seronegative and asymptomatic at birth. Evidences suggests that BVDV contributes to BRD complex potentiating secondary infections caused by Mannheimia haemolytica e Pasteurella multocida due to its immunosuppressive action. However, the farmers have often associated the respiratory syndrome with other infectious agents. This paper reports the attendance of dairy calves manifesting clinical signs of bronchopneumonia, which led to the screening of the persistently infected animals to control of the BVDV infection in the herd.Materials, Methods & Results: During the technical assistance, ten calves manifesting bronchopneumonia were selected to trans-tracheal lavage (TL) in order to identify possible infectious agents. Reverse transcription polymerase chain reaction (RT-PCR) detected the presence of BVDV in two heifers. Pasteurella multocida was the unique bacterial agent isolated from TL (5/10, 50%). These data motivated the technical team and producers to investigate the PI screening by direct enzyme-linked immunosorbent assay from biopsies of the ear edge. The screening of PI’s detected 29 positive within of 2,342 animals tested (1.23%). The re-test of positive was performed only in 24 animals due to the cull of five bovine with severe bronchopneumonia and diarrhea, confirming 18 persistently infected calves (18/24; 75%). Finally, in all PI’s live dams were tested. It was observed four positive adult animals. One grand dam was live and tested, but it had negative result for direct enzyme-linked immunosorbent test. The rate of PI’s considering the whole herd was 0.81% (22/3,700 animals).Discussion: The involvement of BVDV in the etiology of bronchopneumonia was confirmed by detection of the virus in trans-tracheal lavages in two calves by RT-PCR. The susceptibility for Pasteurella multocida infection could be promoted by BVDV prime-infection, considering that immunossupressive nature of BVDV is a critical factor in the interaction with others viruses and bacteria. At this time, we are aware about any report about the detection of BVDV in trans-tracheal lavages. These findings culminated with the screening of PI animals in the herd, detecting rates of 0.81%. The intensive vaccination and colostrum management of this farm could protected the herd against BVDV, however others facts facilitated the introduction of the virus in the herd. This research was conduced in a high-production dairy farm with around 3,700 animals raised in an open herd, in which some of cows with high genetic potential were transferred for embryo collection in the state of Paraná, Brazil; resulting in the addition of the calves to the herd by others routes. Moreover, the farm used for many years vaccine containing only BVDV-1, which may have favored the entry and spread of BVDV-2 or BVDV-3 in the herd. This research showed the presence of BVDV in trans-tracheal lavage of heifers with bronchopneumonia by RT-PCR. This fact points to the need of BRD control programs that include detection of PI animals.

O objetivo desta revisão é trazer informações atualizadas sobre o papel dos fatores imunes do colostro, além das imunoglobulinas (Igs), na proteção direta ou indireta dos bezerros. Os recém-nascidos são hipogamaglobulinêmicos e imunossuprimidos, além de possuírem sistema imune adaptativo näive. Desta forma, é consensual que a colostragem é prioridade no sistema de criação de bezerras, entretanto as novas descobertas sobre a atuação dos leucócitos maternos e seus subprodutos (citocinas), amplificando os mecanismos da resposta imunes inata e adaptativa, tem gerado muitas discussões no meio científico internacional. Além disso, as recentes descobertas sobre o microbioma humano têm sugerido que o processo de amamentação é fundamental para a transferência vertical de Lactobacillus e Bifidobacterium, microrganismos que desempenham um importante papel no desenvolvimento da imunidade de mucosas e saúde intestinal. Em geral, estas descobertas possuem aplicabilidade e trazem reflexões sobre o manejo tradicionalmente empregado na criação de bezerras, considerando-se o contato entre a vaca e recém-nascido, colostragem, sistema de aleitamento, higienização e desinfecção ambiental.

Para a avaliação da transferência de citocinas para o sangue de bezerros neonatos via ingestão de colostro de fêmeas bovinas holandesas, foram utilizados 15 bezerros nascidos de parto eutócico, distribuídos igualmente por três grupos experimentais (n=5): G1- receberam dois litros de colostro fresco provenientes de suas próprias mães; G2- receberam dois litros de colostro provenientes de "pool" de colostro congelado e o G3- foram alimentados apenas com leite. Nestes grupos foram coletadas amostras de sangue em cinco tempos durante os primeiros quinze dias de vida e mensuradas as concentrações das citocinas Interleucina-1 β (IL-1b), Interleucina-6 (IL-6), Fator de necrose tumoral- α (TNF-a) e Interferon-γ (IFN-γ). Também se mensurou tais citocinas (IL-1 β, IL-6 e TNF-α) nos sobrenadantes do colostro de do "pool" de colostro fornecidos aos bezerros dos grupos G1 e G2 respectivamente. Verificou-se a transferência das citocinas IL-1b, IL-6, TNF-a e IFN-γ pela presença no soro dos bezerros do grupo G1, enquanto que nos demais grupos (G2 e G3) não foram detectadas. To evaluate the transfer of cytokines into the blood of neonates via colostrum intake of Holstein cows were used 15 calves born from eutocic, shared equally by three experimental groups (n=5) G1-received 2 liters of fresh colostrum from their own mothers, G2-received 2 liters of colostrum from the "pool" of frozen colostrum and G3-were fed only milk. These group blood samples were collected at five times during the first fifteen days and measured the concentrations of IL-1b, IL-6, TNF-a and IFN-γ. Also measured are the cytokines IL-1 β, IL-6 and TNF-α in the supernatant of colostrum fed to calves from G1 , and cytokines IL-1β , IL-6 , TNF-α and IFN-γ in the supernatant the "pool " of colostrum fed to calves in G2 . It is the transfer of IL - 1b , IL- 6, TNF-a and IFN-γ in the serum of calves from G1 whereas the other groups G2 and G3 were not detected.

O presente trabalho objetivou realizar um estudo retrospectivo sobre os prolapsos vaginal e uterino em ovelhas atendidas no Serviço de Clínica de Bovinos e Pequenos Ruminantes (CBPR) da FMVZ/USP no período compreendido entre 2000 a 2010, no qual, foram atendidas 56 ovinos com problemas inerentes ao sistema reprodutivo, dessas, 25 apresentaram prolapso vaginal ou uterino (44,6%). O prolapso vaginal total foi o de maior frequência (72%). As ovelhas acometidas, em sua maioria, possuíam idade superior a quatro anos (64%), eram sem raça definida (44%) ou da raça Ile de France (40%). As manifestações clínicas observadas durante a maioria dos atendimentos foram: taquipnéia, taquicardia, mucosas oculares avermelhadas indicando estado de toxemia, decúbito esternal ou lateral, apatia e anorexia. O tratamento instituído para todos os casos foi a limpeza, desinfecção e reintrodução do órgão prolapsado. A sutura de Bühner foi feita em 84% dos casos e a histeropexia em um caso (4%). A evolução foi satisfatória em 80% dos casos atendidos, nos demais casos (20%) observou-se óbito da fêmea acometida. Do total de óbitos, os prolapsos vaginais foram responsáveis por 60% (3/5) e os prolapsos uterinos por 40% (2/5). A etiologia dos prolapsos não foi definida nos casos atendidos, sendo esses associados com o período pós-parto em sua maioria (56%), provavelmente associados com quadros de hipocalcemia, altas concentrações séricas de estrógeno e hipertonia uterina. Além disso, a predisposição genética não pode ser descartada.

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 (p0,05) 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.

Milk samples from 45 halves were collected from lactating ewes to evaluate the effect of freezing and incubation of the whole milk on the qualitative results of bacteriologic culture. Thus, these milk samples were submitted to the following treatments: standard culture technique (T1), incubation for 18h at 37°C (T2) and thawing at -20°C for 24h (T3). After these periods, the milk samples from T2 and T3 were submitted to standard culture technique. The T2 showed an increase in the proportion of coagulase-negative staphylococci (CNS) recovery compared to T1, although the same cannot be predicted by T3. Conversely, the T2 allows the growth of some bacteria present in clinically healthy teats ducts as Bacillus spp.. The results of the present study indicated that the incubation of the whole milk can be applied to detection of CNS in ewes’ milk in attempt to reduce the false-negative culture milk samples that can lead to a more efficient use of laboratory resources and reduce costs to herd owners.

A particularidade da secreção láctea caprina, do tipo apócrina, diferente da secreção merócrina da vaca, leva a erros de interpretação durante a realização de técnicas de avaliação da celularidade do leite de fêmeas desta espécie. Portanto, o presente trabalho teve o objetivo de determinar a contagem de células somáticas pelo método indireto California Mastitis Test (CMT), e por métodos diretos, incluindo a contagem por citometria de fluxo e a contagem microscópica direta, através da coloração de verde de metil e pironina-Y, além de comparar os métodos de contagem celular. Foram analisadas 102 amostras de 51 fêmeas caprinas, das raças Saanen, Parda Alpina e Toggenburg, criadas no Estado de São Paulo. Os animais foram categorizados segundo a fase da lactação, exame físico da glândula mamária e exame do leite. As amostras foram colhidas, após a realização do exame Califórnia Mastitis Test, em duas alíquotas, uma destinada à contagem celular automática e a outra, a contagem microscópica direta, utilizando-se o corante verde de metil e pironina- Y. De acordo com os diferentes escores do CMT, observou- se 74,5% de amostras negativas, 8,8% de amostras com escore traços, 8,8% de amostras ligeiramente positivas (+), 6,8% de amostras fracamente positivas (++) e 0,9% de amostras fortemente positivas (+++). Os valores medianos das contagens de células somáticas presentes no leite de cabras, avaliadas através de contador automático e microscopia direta, e analisadas de acordo com os diferentes escores do CMT, foram, respectivamente, 181.000, 578.000, 628.000, 1.421.500 e 5.542.000 células/mL de leite e 74.991, 271.396, 71.420, 640.995 e 5.049.394 células/ mL de leite, nos escores negativo, traços, +, ++ e +++. Os valores medianos obtidos através da contagem de células somáticas pelo método automático e microscópico direto, de acordo com as fases de lactação foram de 159.500, 508.000 e 277.500 células/mL de leite, e 62.493, 89.275 e 146.411. A correlação obtida entre a contagem celular automática e microscópica direta foi de 88%. A partir dos resultados observados pode-se concluir que existe diferença na contagem celular determinada através do método automático e microscópico sendo este último o mais adequado para a determinação da celularidade no leite de cabras. The particular apocrine secretion of goat milk different from the merocrine one observed in cows, may lead to errors in interpreting cellularity evaluations in the milk of this species. Thus, the objective of the present trial was to determine Somatic Cell Counts by means of one indirect methods, the California Mastitis Test (CMT), and direct methods, flow cytometry and direct microscopic count using methyl green-pyronine-Y stain, beyond comparing the methods of cellular counting. A total of 102 samples from 51 Saanen, Brown Alpine and Toggenburg female goats, bred in the state of São Paulo, were analyzed. Goats were separated in groups according to the phase of lactation and to physical examination of the mammary gland, and milk examination. Samples were divided into two aliquots, and were collected after California Mastitis Test evaluation. One aliquot was used in automatic cell counts, and the other, in direct microscopic count using methyl green-pyronine-Y stain. CMT results were as follows: 74.5% of the samples were negative, 8.8% yielded traces, 8.8% were weak positive (1), 6.8% were distinct positive (2) and 0.9% were strong positive (3). Medians of somatic cell counts in goat milk as evaluated by means automatic cell counter and direct microscopy, and grouped according to the different CMT scores, were as follows: 181,000, 578,000, 628,000, 1,421,500, and 5,542,000 cells/mL of milk and 74,991, 271,396, 71,420, 640,995, and 5,049,394 cells/mL of milk in scores negative, traces, 1, 2 and 3, respectively. Medians obtained in automatic cell counts and direct microscopic counts, grouped according to the phase of lactation were 159,500; 508,000; and 277,500 cells/mL of milk and 62,493; 89,275; and 146,411 cells/ml of milk, respectively. The correlation between the automatic and microscopic methods for somatic cell counts was 88%. Based on the results obtained, it could be concluded that there were differences between the automatic and microscopic methods for somatic cell counts, being this most adequate for the determination of the celularidade in the cellularity of goat milk.

Changes in the composition of milk secretion that ocorremdurante the early stages of involution indicate rapid structural changes in the gland and the mechanisms involved in milk synthesis and secretion. Thus, this study sought to elucidate the physicochemical characteristics and cell of milk from sheep of the Santa Ines during the initial period of mammary involution, evaluating the hydrogen ion concentration, somatic cell count (SCC) and the lactose , protein, fat and total solids, in addition to bacteriological examination. In the post-weaning period significant changes were observed in the composition of milk secretion, as increasing pH and protein content, reduced levels of lactose, total solids and fat, plus a trend in increasing CCS. In the positive bacteriological examination animals were also found changes similar to those that occur in the active involution, except fat. Both at the end of breastfeeding period as the period of active involution was exclusively isolated genus Staphylococcus. Changes in the composition of milk secretion that occur during the early stages of involution indicate rapid structural changes in the gland and the mechanisms involved in milk synthesis and secretion. Thus, this study sought to elucidate the physicochemical characteristics and cell of milk from sheep of the Santa Ines during the initial period of mammary involution, evaluating the hydrogen ion concentration, somatic cell count (SCC) and the lactose , protein, fat and total solids, in addition to bacteriological examination. In the post-weaning period significant changes were observed in the composition of milk secretion, as increasing pH and protein content, reduced levels of lactose, total solids and fat, plus a trend in increasing CCS. In the positive bacteriological examination animals were also found changes similar to those that occur in the active involution, except fat. Both at the end of breastfeeding period as the period of active involution was exclusively isolated genus Staphylococcus.

The objectives of the present study were the analysis of cell counts, the comparison between different methods for counting cells and the quantification of different types of cells observed in the milk of Lacaune sheep, using automatic somatic cell counts (SCC) and direct microscopic SCC. Milk smears were stained with methyl green-pyronin-Y and Rosenfeld stain. Mean of automatic cell counts was 0.145 x106 cells/mL of milk, and mean of direct microscopic counts using methyl green-pyronin-Y and Rosenfeld were respectively 0.157 and 0.193 x106 cells /mL of milk. Samples used in direct microscopic counts were evaluated according to the different types of cells and the following results were recorded: 0.094 and 0.14 x106 mononuclear cells /mL, 0.063 and 0.051 x106 polymorphonuclear cells /mL of milk using methyl green-pyronin-Y and Rosenfeld stain, respectively. It may be concluded that there were differences in the methods used for counting cells. The number of cells recorded was lower when methyl green-pyronin-Y and automatic counts were used, compared with direct microscopy using Rosenfeld stain. KEY WORDS: somatic cells and cytology, lacaune, milk, sheep.