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We wished to determine if Mycoplasma bovis infection can negatively impact milk quality and production in Holstein dairy cows. For this Research Communication, milk samples (271) from Holstein cows from 3 herds were screened for M. bovis by real-time PCR. Positive (n = 21) and negative animals (n = 21) were matched by herd, age, lactations and days in milk (DIM). Pairs were evaluated in 7 stages of lactation: D1–50, D51–100, D101–150, D151–200, D201–250, D251–300, and D ≥ 301. A mixed model was used to assess the effect of groups (M. bovis+ × M.bovis−), time (lactation) and groups × time interaction. Cows positive for M. bovis had lower average milk production per day and high somatic cells count (SCC).

The aim of the study was to evaluate the impact of Bovine Leukemia Virus (BLV) on the hematological, biochemical, and acute phase proteins in naturally infected dairy heifers through the transition period. Pregnant Holstein and Jersey heifers (n=24) were distributed into 2 experimental groups: (BLV+) and (BLV-), and BLV+ group was divided into two subgroups presenting high and low proviral load. Animals were assessed at weeks -3, -2, -1, parturition (0), +1, +2, +3 weeks. Blood samples were harvested for RBC, WBC, biochemical exams, including acute phase proteins biomarkers. Herd prevalence of serum positive animals for BLV was 57.25%, and 38.7% in pregnant heifers group. BLV has effect on hematological parameter, in addition to the increase of AST and high concentration of fibrinogen. Also, BLV+ heifers had high risk of inflammation according to the HP cut-off ≥ 2.0 mg/dL (OR= 4.959). Leukocyte counts from animals with high viral load were higher in the high viral load group (P=0.018), especially by lymphocytosis (P=0.036). This is the first publication presenting the impact of BLV on the health of dairy heifers through transition period, reporting a high risk of inflammation. Viral load can impact leukocyte counts, especially by predisposing lymphocytosis.

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The aim of this study was to evaluate the capacity of cells from colostrum to modulate the intestinal microbial colonization, the activity of the inflammatory response, and for their influence on the development of diarrheal disease in calves. Twenty calves were distributed into two groups: COL+ (n = 10) receiving fresh whole colostrum; COL- (n = 10) receiving pooled frozen colostrum, containing no viable cells. All assessments were made before colostrum intake (D0), the next day (D2), and weekly on the 7th (D7), 14th (D14), 21st (D21) and 28th (D28) day of age. Diarrhea was assessed using a fecal score, and the systemic inflammatory status was assessed using a combination of temperature, anemia, total serum iron level, total haptoglobin concentration and the need for systemic antimicrobial treatment. The number of indicator bacteria present in the fecal population was estimated using qPCR. However, COL- calves presented more frequent signs of systemic inflammatory response including, fever at D7 (P = 0.011); indicator haptoglobin levels on D7 and D14, and lower levels of iron on D7, D14. Anemia was detected more often in the COL- calves on D21 (P = 0.043) and D28 (P = 0.016). COL- calves had a 1.66 greater chance of having elevated haptoglobin and a 1.8 greater chance of needing treatment with antimicrobials than COL+. A lower number of DNA copies of Clostridium perfringens were detected in COL+ calves on D2 (P = 0.088) and D7 (P = 0.040). Similarly, a low number of DNA copies was observed for Escherichia coli and Lactobacillus spp. (P = 0.012) in the fecal samples of COL+ calves on D7.

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.

The early use of antimicrobial therapy has been introduced in many farms to prevent diarrhea and respiratory disease in young calves; however, there is controversy about whether this practice has a beneficial effect on the health of these animals. This study evaluated the influence of the early use of antimicrobials on the health and performance of neonatal Holstein calves. Twenty-six Holstein calves were screened and divided into two groups, according to the administration (ATB+), or not (ATB-) of tulathromycin (2.5mg/kg, subcutaneously) within the first 12 hours of life. Calves were evaluated by general clinical examination, fecal score, respiratory score, and external palpation of the umbilical region, besides fecal output of dry matter. Anemia was determined by using an automatic system and, also, using a commercial kit for iron dosage. Diarrhea was diagnosed by a centrifuge-flotation technique using a sugar solution (Cryptosporidium) and multiplex semi-nested RT-PCR (rotavirus/coronavirus). The performance of the calves was estimated by Daily Weight Gain (DWG). The young dairy calves were evaluated within 12 hours of birth (≤12h) and at 3-5th (D3-5), 7-9th (D7-9), 13-15th (D13-15), 20-23rd (D20-23), and 27-30th (D27-30) days of life. No difference was noted between the ATB+ and ATB- groups concerning heart rate, respiratory frequency, and rectal temperature. Erythrogram showed a higher frequency of anemia in ATB- group (P=0.016) at the D3-5 check-up; lower values of serum iron were also observed simultaneously (P=0.051). Thirteen cases of respiratory disease were detected during this study; however, no significant difference was observed between the groups in this regard. The frequency of diarrhea (fecal score 2-3) was high in both groups, peaking at D13-D15. No differences were noted between the groups regarding the frequency of diarrhea when considering the dry fecal matter. The predominant etiological agent for diarrhea was Cryptosporidium spp.. The DWG was similar between groups, with maximum weight reduction on D13-15. The administration of tulathromycin in prophylactic dose (2.5mg/kg) at birth decreased the frequency of anemia but did not influence weight gain or the prevalence of diarrhea.

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.

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

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.

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: 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.

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.

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.

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.

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.

O presente trabalho teve como objetivo investigar a influência do estágio da lactação sobre as características físico-químicas e a celularidade do leite em ovelhas da raça Santa Inês. Desse modo, 143 amostras de leite negativas no exame bacteriológico e sem alterações na prova da caneca de fundo escuro provenientes de 33 ovelhas Santa Inês foram utilizadas para determinar a contagem automática e microscópica de células somáticas, os teores de extrato seco total, gordura, lactose e proteína. Os resultados do presente estudo apontaram para aumento da celularidade do leite e dos teores de gordura e proteína e redução do teor de lactose no final da lactação, embora não tenham sido observadas alterações no extrato seco total no leite desses animais durante a lactação. O presente estudo apontou ainda para correlação significativa entre a contagem automática e a microscópica de células somáticas (r = 0,80), a porcentagem de leucócitos mononucleares (r = -0,45), a porcentagem de leucócitos polimorfonucleares (r = 0,45), os teores de lactose (r = - 0,44), proteína (r = 0,37) e gordura (r = 0,30). No entanto, não se observou correlação significativa entre a CCS automática e o teor de extrato seco total (r = 0,007). The purpose of the present trial was to investigate the influence of lactation stage on milk cellularity and physicochemical characteristics. Thus, 143 milk samples from 33 Santa Ines ewes negative in the bacteriological examination and without any alteration in the strip cup test were used to determine the automatic and microscopic somatic cell count (SCC), and the fat, lactose, protein and total solids content. The results of the present study showed an augment in the SCC and in the fat and protein content, besides a reduction in lactose content at the last lactation stage. Otherwise, no alteration was observed in the total solids content during the lactation period. This study pointed out to a significant correlation between the automatic SCC and the microscopic SCC (r = 0.80), percentage of polymorphonuclear leukocytes (r = 0.45), percentage of mononuclear leukocytes (r = -0.45), lactose (r = - 0.44), protein (r = 0.37) and fat (r = 0.30) content. Nevertheless, significant correlation between the automatic SCC and the total solids content was not observed.

The detection of Bovine Viral Diarrhea virus (BVDV) infection, especially among persistently infected calves (PI), should be performed earlier in order to eliminate the source of the infection and to prevent the spread of the disease in the herd. However, colostrum intake can influence the results of some of the tests used to diagnose the BVDV infection. Therefore, this study evaluated the efficacy of serum neutralization (SN) test in conjunction with reverse transcription-polymerase chain reaction (RT-PCR) for the diagnosis of BVDV infection before colostrum intake. The deliveries of the animals were assisted to select 52 newborn Holstein calves for inclusion in the study. Initially, the whole blood and serum samples were collected from the calves before (T0) and after (T1) the colostrum intake. The calves that were RT-PCR positive at any of the time-points were retested on the 30th day post birth (T2). The presence of specific antibodies for BVDV was evaluated by SN, and that of viral RNA by the RT-PCR. The BVDV-specific antibodies were observed in the serum of 13.46% (7/52) of the calves at T0 because of fetal infection. At T1, seroconversion was observed in 100% (52/52) of the calves. The geometric mean titers (GMT) of the antibodies for BVDV increased significantly from T0 (14.52) to T1 (2490) (P = 0.0001). Of the four calves that were RT-PCR positive before colostrum intake (T0), two were seronegative and two, seropositive. Of the forty-eight RT-PCR negative calves, five were seropositive. After 30 days post birth, all of the animals tested negative by RT-PCR, thus excluding the possibility of persistent infection. The association observed between the results of the SN and RT-PCR assays at T0 (P = 0.025) could not be observed at T1 (P > 0.05). The SN test before the colostrum intake allowed the detection of fetal infection in the herd; however, this test was ineffective as a diagnostic method after the transfer of passive immunity. The confirmation of the results of the SN assay by those of the RT-PCR was essential for the identification of the infected calves before colostrum intake.

Brazilian spotted fever (BSF) is a rickettsiosis very similar to the Rocky Mountain spotted fever also caused by Rickettsia rickettsii, which is sustained in nature in a cycle involving vectors and mammals [1]. The state of Rio de Janeiro, with one of the highest population densities in Brazil, has one of the highest numbers of BSF cases. However, information about the epidemiology of this disease is lacking. The objectives of the present work were to analyse Rickettsia spp. in the host and vectors collected in municipalities of the Vale do Paraiba, where BSF fatal cases were recently recorded.

Foi avaliado o perfil hematológico de bezerros da raça Holandesa durante o primeiro mês de vida. Foram colhidas 208 amostras de sangue total de 26 bezerras(os) do nascimento aos trinta dias de vida. Os valores hematológicos foram determinados por sistema automatizado associado à contagem diferencial dos leucócitos por metodologia manual. Foram detectadas variações nos componentes do hemograma do nascimento aos 30 dias de vida, exceto para os teores de hemoglobina (Hb) e concentração hemoglobínica corpuscular média (CHCM). Os maiores valores do hematócrito, volume corpuscular médio (VCM) e hemoglobina corpuscular média (HCM) foram observados ao nascimento, com decréscimo nos momentos subsequentes. Nos primeiros dias de vida foi observada leucocitose por neutrofilia e eosinopenia e com o avançar da idade houve aumento gradativo dos linfócitos. Com base nos resultados obtidos pode-se concluir que a adaptação dos bezerros no período pós-neonatal foi caracterizada por variações nos componentes do hemograma, observando-se hemoconcentração e padrão leucocitário compatível com resposta aos glicocorticoides até o 4º dia de vida, responsável pela relação neutrófilo-linfócito > 1,0 ao nascimento. The aim of this study was to evaluate the hematological profile of Holstein calves during the first month of life. Blood samples were harvested (n = 208) from 26 calves, from birth until 30 days of life. Hematologic values were determined by an automatic system associated with differential leukocyte count by manual methods. Variations in the erythrogram components were detected from birth up to the 30th day of life, except for hemoglobin (Hb) concentration and Mean Corpuscular Hemoglobin Concentration (MCHC). At birth, higher values were observed for hematocrit, Mean Corpuscular Volume (MCV) and Mean Corpuscular Hemoglobin (MCH) that decreased in subsequent moments. During the first days of life, leukocytosis was found due to neutrophilia and eosinopenia. A gradual increase of lymphocytes with the increase of age was also observed. Finally, the present research showed that the first month of life is a hematological adaptation period. Based on the results, it detected that blood component variations, characterized by hemoconcentration and leukocyte prolife compatible with glucocorticoids response up to the 4th day of life, were responsible for neutrophil lymphocyte ratio > 1.0 at birth.

Objetivo do trabalho foi identificar os agentes etiologicos e fatores de risco para a mastite de cabras leiteiras. Para tanto, foram colhidas duas aliquotas de 257 amostras de leite provenientes de 136 cabras da raca Saanen, oriundas de seis propriedades com sistema de ordenha manual e mecânica, criadas em sistema intensivo, localizadas no estado de Sao Paulo. A primeira aliquota das amostras de leite foi obtida em frascos contendo o conservante bronopol para a Contagem de Celula Somatica (CCS) eletronica, e a segunda em frascos estereis destinada ao exame bacteriologico. Para avaliacao dos fatores de risco, foram aplicados questionarios epidemiologicos preenchidos pela equipe de pesquisa mediante observacoes visuais e inquerito com tratadores e proprietarios. Foram realizadas analises univariada e multivariada, utilizando-se a regressao logistica multipla. Observou-se prevalencia de 28,79% de infeccao mamaria com predominio de Staphylococcus coagulase negativo (SCN). A mediana da CCS obtida foi de 0,47x10 6 celulas/mL de leite. Na analise univariada considerando-se as variaveis relacionadas no quadro 03 que apresentaram P

A eficiência na transferência de imunidade passiva (TIP) está diretamente associada aos índices de morbidade e mortalidade de bezerros. Quando o colostro materno fresco (CF) não está disponível o colostro congelado (CC) ou o substituto de colostro (SC) são alternativas para evitar falhas na TIP (FTIP). O objetivo do presente estudo foi avaliar a utilização dessas diferentes fontes de colostro e seus efeitos na TIP, saúde, desempenho e metabolismo de bezerros Holandês. Foram utilizados 39 bezerros (machos e fêmeas) em delineamento de blocos casualizados de acordo com sexo, peso e data de nascimento e foram distribuídos em três grupos experimentais: 1) 10% PV em CF de alta qualidade; 2) 10% PV em CC de alta qualidade; 3) 1,5 doses de SC (705 g). Foram colhidas amostras de sangue ao nascimento, antes do fornecimento do colostro (0h) e com 12, 24 e 48h de vida para avaliar TIP e metabólitos. O período experimental foi de 63 dias. Os animais tiveram acesso ad libitum à água e concentrado durante todo o período e até os 56 dias receberam 6L/d de leite. Diariamente foi mensurado o consumo de concentrado, monitorada a saúde dos animais e a incidência de diarreia por meio do escore fecal. Semanalmente avaliou-se peso e medidas corporais, além de coleta de sangue para determinação de proteína total, albumina e glicose. A ingestão de imunoglobulinas foi maior (P < 0,01) para os animais que receberam colostro materno (CF: 304,77 g e CC: 292,57 g) quando comparados com aqueles que receberam SC (150 g). Isso influenciou (P < 0,01) a concentração sérica de IgG com 24 horas. Entretanto, apesar das diferenças apenas um bezerro apresentou FTIP (IgG sérica < 10 g/L). Na contagem diferencial de leucócitos foi observado efeito de interação da fonte de colostro e horário da coleta para os valores de linfócitos (P = 0,01) e eosinófilos (P = 0,03). Dos parâmetros sanguíneos avaliados nas primeiras 48 horas de vida, apenas os valores de insulina foram afetados (P = 0,01) pelas fontes de colostro, apresentando-se elevado nos animais alimentados com SC. O número médio de episódios de doença também foi influenciado (P = 0,02) pelas diferentes fontes de colostro de maneira que os animais que receberam SC apresentaram maior número de quadros de doença do que os animais que receberam CF ou CC (1,77 vs 0,65 e 0,59 episódios respectivamente). O GMD (CF: 613 CC: 551 e SC: 473 g/dia; P = 0,01) e a eficiência alimentar (CF: 0,688 CC: 0,626 e SC: 0,553 kg ganho/kg consumo; P = 0,005) diferiram entre os grupos experimentais, consequentemente, influenciando (P = 0,04) também o peso corporal ao final do experimento (CF: 71,17 CC: 65,03 e SC:64,36 kg). Parâmetros metabólicos como proteína sérica total (P < 0,01) e glicose (P = 0,02) também diferiram de acordo com o colostro ofertado. Concluímos que todas as fontes de colostro utilizadas neste experimento foram eficientes na TIP, entretanto protocolos de colostragem que possibilitem a ingestão de maiores quantidades de imunoglobulinas, proporcionam concentrações séricas de IgG mais elevados, melhores condições de saúde e consequentemente maiores ganho e desempenho. Efficiency in passive immunity transfer (PIT) is directly associated with calf morbidity and mortality rates. When fresh maternal colostrum (FMC) is not available, frozen colostrum (FC) or colostrum replacer (CR) is an alternative to avoid failures in PIT (FPIT). The objective of the present study was to evaluate the use of these different sources of colostrum and their effects in PIT, health, performance and metabolism of Holstein calves. Therefore, 39 calves (males and females) were allocated in randomized blocks according to sex, weight and birth date, and were distributed into three experimental groups: 1) 10% of body weight (BW) in high quality FMC; 2) 10% of BW in high quality FC; 3) 1.5 doses of CR (705 g). Blood samples were collected at birth, before colostrum fed (0h), and with 12, 24 and 48h of life to assess PIT and metabolites. The experimental period was 63 days. All animals had ad libitum access to water and received starter throughout the experimental period and received 6L/d of milk until 56 days of age. Starter intake was measured daily and animals\' health, including incidence of diarrhea, was monitored through fecal score. Weight and body measurements were measured weekly, and blood was collected to determine concentration of total protein, albumin and glucose. Immunoglobulins\' intake was higher (P < 0.01) for the animals that received maternal colostrum (FMC: 304.77 g and FC: 292.57 g) when compared to those that received CR (150 g). This influenced (P < 0.01) the serum IgG concentration at 24 hours. However, despite the differences, only one calf had FPIT (serum IgG < 10 g/L). The differential leukocyte count showed an interaction effect of the colostrum source and time of collection for the values of lymphocytes (P = 0.01) and eosinophils (P = 0.03). Considering blood parameters in the first 48 hours of life, only insulin values were affected (P = 0.01) by the sources of colostrum, being elevated in animals fed with CR. The average number of disease episodes was also influenced (P = 0.02) by the different sources of colostrum so that the animals that received CR had a higher number of illnesses than the animals that received FMC or FC (1.77 vs 0.65 and 0.59 episodes respectively). ADG (FMC: 613 FC: 551 and CR: 473 g/day; P = 0.01) and feed efficiency (FMC: 0.688 FC: 0.626 and CR: 0.553 kg gain/kg consumption; P = 0.005) differed between the experimental groups, consequently, influencing (P = 0.04) also the body weight at the end of the experiment (FMC: 71.17 FC: 65.03 and CR: 64.36 kg). Metabolic parameters such as total serum protein (P < 0.01) and glucose (P = 0.02) also differed according to the colostrum offered. In conclusion, although all sources of colostrum resulted in efficient PIT, those that allowed ingestion of greater amounts of immunoglobulins provided higher serum IgG concentrations, better health conditions and, consequently, greater gain and performance.

Em teoria, bezerros leiteiros nascem com o trato gastrintestinal estéril e durante o nascimento, inicia-se a colonização microbiana. A partir desse momento, a dieta torna-se um importante fator de manutenção da população microbiana, além de servir de substrato para a mesma. No entanto, pouco se sabe como diferentes dietas podem modificar o microbioma gastrintestinal em bezerros leiteiros durante a fase de aleitamento e desaleitamento. Com base nisso, dois estudos foram realizados. O primeiro estudou avaliou diferentes níveis e fontes de FDN no concentrado inicial. Foram utilizados 18 bezerros Holandeses, alimentados com um dos três tratamentos: 22FDN - concentrado convencional contendo 22% de FDN; 31FDN - concentrado com 31% de FDN, substituindo parte do milho por casca de soja; e 22Feno - concentrado convencional com 22% de FDN e feno coast cross a vontade. Foram alojados em gaiolas suspensas, sem cama até a 8ª semana de vida, desaleitados e depois alojados em abrigos do tipo tropical até a 10ª semana de vida. Foram avaliados o microbioma ruminal, nas semanas 2, 4, 6, 8 e 10, e o microbioma fecal nas semanas 0, 1, 2, 4, 8 e 10. O microbioma ruminal não mostrou diferenças em função da dieta, idade ou interação de ambos os fatores. O microbioma fecal foi afetado tanto pela idade quanto pela dieta. A inclusão de feno na dieta pode promover maior diversidade microbiana fecal. O segundo estudou avaliou diferentes dietas líquidas durante o aleitamento. Foram utilizados 15 bezerros Holandeses alimentados com uma das três dietas líquidas: leite integral, sucedâneo lácteo, e leite integral acidificado a pH 4,5 com ácido fórmico. Os animais foram alojados em abrigos tropicais, recebendo 6 litros de dieta líquida por dia, sendo desaleitados a partir da 8ª semana de vida, e alojados em abrigo coletivo a partir do desaleitamento. O microbioma fecal foi avaliado nas semanas 0, 1, 2, 4, 8 e 10. A diversidade microbiana foi afetada pela idade e dieta. A diversidade foi crescente a partir da semana 1. O leite promoveu maior diversidade microbiana fecal em comparação as demais dietas, além de promover o crescimento de bacterias benéficas e comensais no instetino. No entanto, após os animais deixarem de receber as dietas líquidas e serem alojados em conjunto, o microbioma fecal desses animais convergiu para uma maior similaridade. In theory, dairy calves are born with a sterile gastrointestinal tract and at birth, microbial colonization begins. From that moment on, the diet becomes an important factor for maintaining the microbial population, in addition to serving as a substrate for growth. However, it is necessary to know how different diets can modify the gastrointestinal microbiome in dairy calves during the pre-weaning and weaning phase. Based on that, two studies were performed. The first study evaluated different levels and sources of NDF in the starter concentrate. Eighteen Holstein calves were used, fed with one of three treatments: 22NDF - conventional starter concentrate containing 22% NDF; 31NDF - starter concentrated with 31% NDF, replacing part of the corn with soybean hull; and 22Hay - conventional concentrate with 22% NDF and coast cross hay at will. The animals were housed in suspended not-bedded cages, until the 8th week of life, weaned and then housed in tropical shelters until the 10th week of life. The rumen microbiome was evaluated at weeks 2, 4, 6, 8 and 10, and the fecal microbiome at weeks 0, 1, 2, 4, 8, and 10. The ruminal microbiome had no differences depending on diet, age, or interaction of both factors. The fecal microbiome was affected by both age and diet. The supply of hay in the diet can promote greater fecal microbial diversity. The second studied evaluated different liquid diets during the pre-weaning phase. Fifteen Holstein calves fed one of the three liquid diets were used: whole milk, milk replacer, and whole milk acidified to pH 4.5 with formic acid. The animals were housed in tropical shelters, receiving 6 liters of liquid diet per day, being weaned after the 8th week of life, and housed in a collective shelter from weaning. The fecal microbiome was evaluated at weeks 0, 1, 2, 4, 8, and 10. Microbial diversity was affected by age and diet. Diversity increased from week 1. Whole milk promoted greater fecal microbial diversity compared to other diets, in addition to promoting the growth of beneficial and commensal bacteria in the feces. However, after the animals stopped receiving liquid diets and were housed together, the fecal microbiome of these animals converged towards greater similarity.

O objetivo do presente estudo foi avaliar a transferência de imunidade passiva, saúde e o desempenho de bezerros colostrados com diferentes doses de imunoglobulinas provenientes do substituto de colostro SCCL® ou colostro materno. Os animais foram separados das mães imediatamente após o nascimento. Cinquenta bezerros da raça holandesa (machos e fêmeas) foram alocados em blocos casualizados de acordo com sexo, peso ao nascer e data de nascimento e distribuídos em diferentes protocolos de colostragem: 1) 2L de colostro materno (2CM); 2) 4L de colostro materno (4CM); 3) 2L de colostro materno + uma dose do substituto de colostro (2MC1SC); 4) 2 doses substituto de colostro ao nascer (2SC); e 5) 2 doses substituto de colostro ao nascer + uma dose substituto de colostro entre 6 a 8 horas após nascimento (3SC). Foram colhidas amostras de sangue ao nascimento, antes do fornecimento do colostro (0h) e com 48h após a ingestão do colostro para avaliar a transferência de imunidade passiva. Durante o período de aleitamento os bezerros receberam 6L/d de sucedâneo, tiveram acesso ad libitum à água e concentrado e foram alojados em abrigos individuais, tipo tropical. O consumo de concentrado foi avaliado diariamente, e semanalmente avaliou-se o peso e medidas corporais. A saúde dos animais foi monitorada diariamente e a incidência de diarreia foi avaliada através do escore fecal. Durante o aleitamento as amostras de sangue foram colhidas a cada 14 dias, para determinação de proteína total, albumina, glicose, BHBA, AGNE e insulina. A transferência de imunidade passiva não foi afetada pelos protocolos de colostragem (P>0,05). No entanto, doses mais elevadas de colostro resultaram em maior ingestão de imunoglobulinas, porém reduz a eficiência aparente de absorção (P 0,005). No entanto, afetou o número de tratamentos contra tristeza parasitária bovina (P=0,0042). O peso médio e o GMD, assim como os parâmetros sanguíneos avaliados durante o aleitamento, não foram afetados pelos protocolos de colostragem (P> 0,05). O substituto de colostro avaliado no presente estudo pode ser uma alternativa ao colostro materno, uma vez que, tanto o colostro materno como o substituto de colostro foram eficientes na transferência de imunidade passiva, resultando em desempenho similar mesmo em condições de alto desafio sanitário. The objective of the present study was to evaluate the passive transfer of immunity (PTI), health and performance of calves fed different doses of immunoglobulins from the SCCL® colostrum replacer or maternal colostrum. The animals were separated from the mothers immediately after birth. Fifty holstein calves (male and female) were allocated in randomized blocks according to sex, birth weight and date of birth and were distributed among different colostrum feeding protocols: 1) 2L of maternal colostrum (2MC); 2) 4L of maternal colostrum (4MC); 3) 2L of maternal colostrum + one dose of colostrum replacer (1MC+1CR); 4) 2 doses of colostrum replacer (2CR) and 5) 2 doses of colostrum replacer given at birth + one dose of colostrum replacer given between 6 and 8 hours after birth (3CR). Blood samples were taken at birth, before colostrum fed (0h) and at 48h after colostrum intake to assess passive immunity transfer. Calves were individually housed and bucket fed 6L/d of milk replacer, and had ad libitum access to water and starter. Starter intake was measured daily, and weekly assessed weight and body measurements. Animal health was monitored daily and the incidence of diarrhea was assessed by fecal score. During the pre-weaning, blood samples were harvested every 14 days to for determination of total protein, albumin, glucose, BHBA, NEFA and insulin. The passive transfer of immunity was not affected by colostrum feeding protocols (P>0.05). However, higher doses of colostrum resulted in higher immunoglobulins intake, but lower apparent efficiency absorption (P0.05), such as fecal score (P=0,5873), days with diarrhea (P=0,3685), days with fever (P=0,7072), number of treatments for diarrhea (P=0,8499) and number of treatments for pneumonia (P>0,005). However, affected the number of treatments for cattle tick fever (P=0.0042). The mean weight and ADG, as well as the blood parameters evaluated during pre-weaning were not affected by the colostrum feeding protocols (P> 0.05).The colostrum replacer evaluated in this study may be an alternative to maternal colostrum, once the, both maternal colostrum and colostrum replacer were efficient in the transfer of passive immunity, resulting in similar performance even under conditions of high sanitary challenge.

O objetivo deste trabalho foi identificar o padrão de vigor e status metabólico através da determinação da frequência dos sinais objetivos e metabólitos sanguíneos após o nascimento, possivelmente associados à transferência de imunidade passiva, além de determinar o melhor horário para o uso do refratômetro de Brix na avaliação da transferência de imunidade passiva em bezerros neonatos após a ingestão de colostro. No primeiro experimento foram utilizados 46 bezerros neonatos de ambos os sexos, nascidos entre setembro de 2013 e julho de 2014. A avaliação do vigor dos animais foi realizada entre 15 e 30 minutos após o nascimento com o auxílio da escala APGAR modificada pela Universidade de Guelph. A colheita de sangue para análise de metabólitos seguiu os horários de 0h (antes da ingestão), 1h, 2h, 4h, 6h, 12h, 24h, 48h contadas a partir da ingestão do colostro. Os parâmetros sanguíneos foram determinados por kit enzimático específico para cada parâmetro em Sistema Automático para Bioquímica. Todos os animais utilizados neste estudo foram oriundos de partos eutócicos e apresentaram, de acordo com a escala APGAR, vigor satisfatório. As concentrações de metabólicos sanguíneos dos neonatos mostraram que o consumo de colostro aumentou a disponibilidade de energia e a concentração de frações proteicas, comumente utilizadas como indicativos da transferência de imunidade passiva. No segundo experimento foram utilizados 47 bezerros neonatos de ambos os sexos, nascidos entre março e julho de 2014. O colostro foi ordenhado após o parto e a qualidade determinada com auxílio do colostrômetro e do refratômetro digital. A colheita do sangue para a análise dos parâmetros sanguíneos seguiu os horários de 0h (antes da ingestão), 1h, 2h, 4h, 6h, 12h, 24h, 48h, 72h, 96 e 120h, contadas a partir da ingestão do colostro. A concentração de proteína total foi determinada por refratômetro de Brix e por kit enzimático em Sistema Automático para Bioquímica. Os demais parâmetros sanguíneos foram determinados por kit enzimático específico para cada parâmetro em Sistema Automático para Bioquímica. A contagem global e diferenciada das células sanguíneas foi realizada nos tempos 0, 12 e 24h após o fornecimento do colostro. O colostro se manteve na faixa de boa qualidade, segundo os critérios de classificação de medidas pelo colostrômetro e refratômetro digital de Brix. O consumo de colostro foi o principal fator de variação dos parâmetros estudados neste trabalho. A avaliação da transferência de imunidade passiva em bezerros recém-nascidos deve ser realizada por volta das 24 horas após a ingestão do colostro, pois neste momento a absorção de macromoléculas colostrais já está encerrada e a PT alcança estabilidade. A partir deste período, não é possível determinar se as frações proteicas avaliadas são de origem colostral ou endógena, o que não garante avaliação segura sobre a transferência de imunidade passiva. The objective of this study was to identify the vigor and metabolic status through the determination of objective signs and blood metabolites just after birth of dairy calves, possibly associated with the passive immunity transfer. Also, determine the best time after colostrum intake to estimate the passive immunity transfer using a Brix refractometer. The first trial evaluated 46 newborn calves, males and females, born from September 2014 to July 2014. Animals\' vigor evaluations were carried out between 15 and 30 minutes after birth using the APGAR scale modified by the University of Guelph. The blood sample collection for metabolites analysis were done at 0h (before intake), 1h, 2h, 4h, 6h, 12h, 24h, and 48h after colostrum intake. Blood parameters were analyzed by enzymatic kits using an automatic biochemical system. All animals were born from eutocic births and presented, according to the APGAR scale, satisfactory vigor. The newborns metabolites blood concentrations revealed that colostrum intake increased the energy availability and the concentration of protein fractions, commonly used as indicative of passive immunity transfer. For the second trial 47 newborn calves of both sexes, born from March and July 2014, were used. The colostrum was milked after parturition and quality was determined using a colostrometer and digital Brix refractometer. Blood samples were taken from calves at 0h (before intake), 1h, 2h, 4h, 6h, 12h, 24h, 48h, 72h, 120h and 96, after colostrum intake. The total protein (TP) concentration was measured by a Brix refractometer and an enzymatic kit using an automatic biochemical system. Other blood parameters were determined by specific enzyme kit for each parameter in Automatic System for Biochemistry. The global and differential blood cell count were performed at times 0, 12 and 24h after colostrum feeding. Colostrum has remained in the range of good quality according to the criteria of classification measures by colostrometer and digital Brix refractometer. Colostrum intake was the main factor of variation of the studied parameters. The evaluation of the passive transfer of immunity to newborn calves should be performed at about 24 hours after colostrum intake, because at this time, the absorption of colostral macromolecules is already finished and TP reaches stability. From this period, it is not possible to determine whether the assessed protein fractions are from colostrum or endogenous origin, which does not guarantee reliable assessment on the transfer of passive immunity.

O objetivo geral deste trabalho foi avaliar a influência dos leucócitos do colostro bovino na imunidade inata, desenvolvimento da microbiota intestinal e ocorrência de diarreias em bezerras Holandesas recém-nascidas. Para isso, 20 bezerras Holandesas foram acompanhadas nos seguintes momentos: antes da mamada do colostro (D0); 1-2 (D2); 7 (D7); 14 (D14); 21 (D21) e 28 (D28) dias após nascimento e foram distribuídas em dois grupos experimentais: grupo COL+ recebeu colostro fresco (4L) proveniente de suas respectivas mães; e grupo COL- recebeu colostro congelado e acelular (4L) de vacas doadoras. Capítulo 1 O objetivo deste capítulo foi avaliar a presença e influência dos leucócitos do colostro na colonização do trato gastrintestinal. Para isso, amostras de colostro foram semeadas em TSA e MacConkey para CBT e CCT; e amostras de fezes em agar sangue, MacConkey e Salmonella-Shiguella. Bezerras apresentaram maior frequência de diarreia no D14 (COL+=78%; COL-=60%). O grupo COL- apresentou maior CBT e CCT/mL (3,93x106 ufc/mL e 3,01x105 ufc/mL) em relação ao COL+ (0,94x106 ufc/mL e 0,78x105ufc/mL). Sobre as espécies bacterianas isoladas, as que foram mais frequentes no COL+ foram Proteus mirabilis (29,91%), Escherichia coli (28,04%), Citrobacter freundii (5,61%) e Staphylococcus spp (5,61%), no COL- foram Escherichia coli (28,70%), Proteus mirabilis (27,78%), Klebsiella pneumoniae (11,32%) e Morganella morganii (5,66%). O momento com maior isolamento em ambos os grupos foi o D2 (COL+ = 26 cepas; COL- = 27 cepas), período em que iniciou a diarreia nas bezerras. Não foi possível detectar diferenças entre as frequências de microrganismos entre os grupos COL+ e COL-. A administração de colostro COL+ e COL- não influenciou na proporção de bactérias aeróbias presentes nas fezes e na ocorrência de diarreia das bezerras durante o período neonatal. Capítulo 2 O objetivo deste capítulo foi avaliar a influência dos leucócitos do colostro na resposta imune inata em bezerras Holandesas. Os animais foram submetidos ao exame clínico, seguido da colheita das amostras sanguíneas para realização das provas laboratoriais. As concentrações de cortisol sérico e haptoglobina não apresentaram diferenças estatísticas, enquanto a concentração de ferro foi diferente entre os grupos no D7. No leucograma não foi detectada diferença para os leucócitos totais e neutrófilos absolutos, mas houve tendência entre os grupos no D7 para os valores de neutrófilos relativos. O marcador CH138+CD62L+ não apresentou diferença entre os grupos, apesar da maior expressão ter sido detectada no COL-. A expressão de CH138+CD62L- foi maior no COL+ no D14. A fagocitose e produção de H2O2 pelos neutrófilos sanguíneos apresentou perfil semelhante em relação ao S. aureus e E. coli entre os grupos, apesar do COL- apresentar maior intensidade de fagocitose do D7 ao D14. Houve tendência para o aumento da proporção de granulócitos liberando H2O2 basal no D14 para o COL-, entretanto o COL + apresentou tendência para maior intensidade da fagocitose no D21. A proporção de granulócitos (%), estimulados com S. aureus, liberando H2O2 foi maior no COL+ em D7, enquanto o estímulo com a E. coli não resultou em diferenças entre os grupos. A intensidade de fluorescência foi maior e gradual no COL+ quando as células foram estimuladas, porém não foram encontradas diferenças estatísticas. As células do colostro influenciaram no perfil sanitário apresentado pelas bezerras, observando-se maior intensidade de diarreias, menores teores de ferro sérico e anemias no grupo que recebeu colostro congelado. A migração dos granulócitos sanguíneos foi mais rápida e intensa no COL+ em relação ao COL-, após exposição natural aos patógenos causadores de diarreia. O índice de fagocitose (%) dos granulócitos apresentou semelhança entre os grupos, entretanto a intensidade da fluorescência foi mais intensa no COL-. Em contrapartida, o COL+ demonstrou maior habilidade em produzir H2O2 The aim of this study was to evaluate the influence of bovine colostrum leukocytes in innate immunity, in the development of the intestinal microbiota and the occurrence of diarrhea in newborn Holstein calves. For this, 20 Holstein calves were followed in the following moments: before colostrum intake (D0); 1-2 (D2); 7 (D7); 14 (D14); 21 (D21) and 28 (D28) days after birth. They were divided into two experimental groups: COL+ group, which received fresh colostrum (4L) from their mothers; and COL-group which received frozen and acellular colostrum (4L) of donor cows. Chapter 1 -The aim of this chapter was to evaluate the presence and influence of colostrum leukocytes in the colonization of the gastrointestinal tract. For this, colostrum samples were seeded in TSA and MacConkey for TPC and TCC; and stool samples on blood agar, MacConkey and Salmonella-Shiguella. Calves presented higher frequency of diarrhea in D14 (COL+ = 78%; COL- = 60%). The COL- group showed higher TPC and TCC/mL (3,93x106 cfu/mL and 3,01x105 cfu/mL) compared to COL+ (0,94x106 cfu/mL and 0,78x105 cfu/mL). Regarding the isolated bacterial species, those that were more frequent in the COL+ were Proteus mirabilis (29.91%), Escherichia coli (28.04%), Citrobacter freundii (5.61%) and Staphylococcus spp (5.61%); in the COL-were Escherichia coli (28.70%), Proteus mirabilis (27.78%), Klebsiella pneumoniae (11.32%) and Morganella morganii (5.66%). The moment with more insolation in both groups was the D2 (COL+ = 26 strains; COL- = 27 strains), period in which started the diarrhea in calves. It was not possible to detect differences between the frequencies of microorganisms between the COL+ and COL- groups. The COL+ and COL- colostrum management did not influence the proportion of aerobic bacteria present in feces and the occurrence of diarrhea in calves during the neonatal period. Chapter 2 The aim of this chapter was to evaluate the influence of colostrum leukocytes in the innate immune response in Holstein calves. The animals were submitted to clinical examination, followed by blood samples collection to perform laboratory tests. The serum cortisol and haptoglobin concentrations did not show statistical differences while the iron concentration was different between groups at D7. The leukogram did not show difference to total leukocytes and absolute neutrophils, but there was a trend between groups at D7 for relative values of neutrophils. The CH138+CD62L+ marker showed no difference between groups, despite an increased expression was detected in COL-. The expression of CH138+CD62L- was greater in COL+ at D14. The phagocytosis and production of H2O2 by blood neutrophils showed similar profile in relation to S. aureus and E. coli between groups, although COL- presented greater phagocytosis intensity from D7 to D14. There was a trend for the increase of granulocyte proportion, releasing basal H2O2 at D14 for COL-, however COL+ presented trend to a higher intensity of phagocytosis at D21. The proportion of granulocytes (%) stimulated with S. aureus releasing H2O2 was greater in COL+ at D7, while stimulation with E. coli resulted in no differences between the groups. The fluorescence intensity was higher and gradual in COL+ when cells were stimulated; however, it was not detected statistical differences. The colostrum cells influenced in the health profile presented by the calves, with a higher intensity of diarrhea, lower levels of serum iron and anemia in the group that received frozen colostrum. The migration of blood granulocytes was faster and more intense in COL+ in relation to COL- after natural exposure to pathogens that cause diarrhea. The phagocytosis rate (%) of granulocytes showed similarity between groups, although the fluorescence intensity of COL- was more intense. In contrast, the COL+ demonstrated greater ability in producing H2O2

O objetivo deste estudo foi avaliar o efeito do suplemento de colostro (SC) associado ao colostro materno (CM) de alta ou média qualidade na saúde e desempenho de bezerros leiteiros. No primeiro experimento foram utilizados 44 machos da raça Holandesa, distribuídos nos tratamentos: 1) CM de alta qualidade fornecido em volume correspondente a 15% do peso ao nascer (PN); 2) CM de média qualidade (15%PN) e 3) CM de média qualidade (15% PN) + SC. O protocolo de colostragem afetou a concentração de proteína total nas primeiras 48 horas de vida e durante a fase de aleitamento (P0,05); no entanto, os animais que receberam CM de alta qualidade foram medicados por um menor número de dias (P0,05), embora tenha ocorrido efeito de idade (P0,05). Os parâmetros sanguíneos nas primeiras 24 horas não foram afetados pelos tratamentos (P>0,05). O consumo e o desempenho não foram afetados pela administração do suplemento de colostro (P>0,05), no entanto houve efeito de idade para ambos os parâmetros (P0,05). A taxa de mortalidade observada durante o experimento foi de 2,5%. O consumo adicional de Ig através do fornecimento de suplemento de colostro em associação ao colostro materno de alta qualidade não melhorou a imunidade, bem como não afetou o desempenho ou a saúde de bezerros leiteiros. No terceiro experimento, foram utilizadas 67 fêmeas da raça Holandesa ou mestiças Girolando nos seguintes tratamentos: 1) CM de alta qualidade fornecido em volume correspondente a 10% PN; e 2) CM de alta qualidade (10% PN) + SC. O consumo e o desempenho não foram afetados pelo SC (P>0,05). O escore fecal também não foi afetado (P>0,05), mas observou-se maior frequência de animais com escore de desidratação nas 2ª e 3ª semana de vida, quando os escores fecais foram mais altos para todos os tratamentos. Houve uma maior porcentagem de animais suplementados sendo medicados devido a diarreias na segunda semana de vida. Houve maior frequência dos animais do grupo não suplementado com escore 1 de descarga nasal nas semanas 3 a 7. O consumo adicional de imunoglobulinas através do fornecimento de suplemento de colostro em associação ao colostro materno de alta qualidade não afetou o desempenho ou a saúde dos animais. The aim of this study was to evaluate the effect of colostrum supplement (CS) associated with maternal colostrum (MC) of high and medium quality on health and performance of calves. In the first experiment 44 Holstein males were used in the following treatments: 1) high quality MC provided in a volume corresponding to 15% of birth weight (BW); 2) medium quality MC (15% BW) and 3) medium quality MC (15% BW) + CS. Colostrum feeding protocol affected the total protein concentration in the first 48 hours of life (P0.05); however, animals that received high quality colostrum were treated for a shorter number of days (P0.05) and were increased over the weeks (P 0.05). Blood parameters during the first 24h were not affected by treatments (P>0.05). The concentrate intake and performance were not affected by the administration of CS (P> 0.05), however there was an age effect (P 0.05). The mortality rate observed during the experiment was 2.5%. The additional intake of Ig via CS in combination with high quality MC have not improved immunity or affected the performance and health of dairy calves. In the third trial, 67 female Holstein or Holstein x Gir were assigned to two treatments: 1) high quality MC supplied in c volume corresponding to 10% BW; and 2) high quality MC (10% BW) + CS. Concentrate intake and performance were not affected by feeding a CS (P> 0.05). The fecal score was not affected by the CS (P> 0.05), but there was a higher frequency of animals with dehydration score during the 2nd and 3rd week of life, when the fecal scores were higher for both treatments. There was a higher percentage of supplemented animals being medicated because of diarrhea during the second week of life. There was a higher frequency of animals in the non-supplemented group with nasal discharge score 1 in weeks 3 to 7. The additional consumption of immunoglobulins through the colostrum supplement feeding in combination with high quality colostrum did not affect the performance or the health of animals.