Search

Simple Summary Parturition is the most important physiological event in the lifecycle of dairy cows; it mediates changes in the microbiota composition. However, the complete picture of the dynamics of these phenomena and how they affect health and metabolism is unknown. This study documents the composition of the microbiota in the mammary gland, on reproductive surfaces and those associated with the rectum immediately after parturition. The microbiomes of different maternal niches were different, as predicted by their different functional roles in cows. Based on the results of this research, the conclusion that the microorganisms that colonize different mucosal tissues of cows were linked to the state of systemic energy metabolism and had an impact on the health of the mammary gland cows following calving was drawn. Abstract During the period called “transition”, from the ceasing of milk production to the reestablishment of full milk production, it is postulated that the microbiota of cows undergo changes in composition driven by the fluxes in systemic energetics and that these changes appear to impact the health of cows. The primary objective of this study was to document the make-up of the microbiota in the mammary gland compared with those in the vagina and in feces in an attempt to determine any correlations between the composition of the microbiota, the impact of blood indicators of energetic metabolites and the health of the mammary gland at the time of calving. Samples were collected from 20 Holstein dairy cows immediately following calving to assess their general health and measure the microbiomes associated with each cow using 16S rRNA sequencing. The results indicated that the microbiomes found within each maternal niche were different. A set of significant negative associations between the blood energetic biomarkers (NEFAs, BHB, triglycerides and cholesterol) and the taxa Pseudomonas, Christensenellaceae and Methanobrevibacter were observed in this study. In contrast, Escherichia and Romboutsia were positively correlated with the same energetic metabolites. Therefore, it was concluded that there appears to be a set of relationships between the microorganisms that colonize several niches of cows and the sufficiency of systemic energy metabolism. Furthermore, both the microbiome and energy dynamics impact the health of the mammary gland of the host.

The present study evaluated the use of haptoglobin (Hp) as an indicator of health and performance in 166 Holstein heifer calves reared in an intensive production system. Calves were evaluated at D6-9; D10-13; D20-23; D35-38 and D65-68, corresponding to the days of life. The absence or presence of diseases was evaluated by physical examination and classification of scores. The performance parameters evaluated were body weight, height at withers and hind width. Hp was measured by spectrophotometric technique. The highest prevalence of diarrhea (59.4%; 98/165) was observed in D10-13, bovine respiratory disease (BRD) was on D35-38 (25.8%; 42/163), and umbilical inflammations in D6-D9 (7.8%; 13/166). Highest values of Hp were observed in animals with diarrhea (P=0.02), and umbilical inflammation (P=0.057), in comparison with the group of healthy calves. A significant negative correlation was observed between Hp and performance index. This protein presented an important relation with diarrhea and performance of the calves, opening perspectives on its utilization as a biomarker of diseases. RESUMO O presente estudo avaliou o uso da haptoglobina (Hp) como indicadora de sanidade e desempenho em 166 bezerras Holandesas criadas em um sistema de produção intensivo. As bezerras foram avaliadas nos momentos D6-9; D10-13; D20-23; D35-38 e D65-68, sendo estes correspondentes aos dias de vida. A ausência ou a presença de doenças foi avaliada por meio do exame físico e da classificação por escores. Os parâmetros de desempenho avaliados foram peso corporal, altura de cernelha e largura de garupa. A Hp foi mensurada por técnica espectrofotométrica. A maior prevalência de diarreia (59,4%; 98/165) foi observada em D10-13, doença respiratória bovina (DRB) ocorreu em D35-38 (25,8%; 42/163) e inflamações umbilicais em D6-D9 (7,8%; 13/166). O valor de Hp foi maior nos animais que apresentaram diarreia (P=0,02) e inflamações umbilicais (P=0,057), em comparação ao grupo de bezerras saudáveis. Houve correlação negativa significativa entre a Hp e os índices de desempenho. Essa proteína apresentou uma importante relação com a diarreia e com o desempenho das bezerras, abrindo perspectivas sobre a sua utilização como biomarcadora de doenças.

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

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

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.

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.

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

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

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

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

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

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

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

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

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

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.

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

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

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

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 measurement of pH in milk is one of the most important and frequent procedures performed in biochemistry practices. This variable is applied to the diagnosis of mammary gland disease, assessment of industrial yields, and identification of metabolic diseases. The objective of this study was to evaluate de effect of freezing the milk from healthy and infected mammary glands of Santa Ines ewes on the pH. The bacteriological culture of 183 milk samples was analyzed resulting in 146 negative and 37 positive samples. From these, the hydrogen ion concentration was determined on sampling day and after freezing at -20°C for 30 months. A reduction on pH was observed after freezing, and the bacteriological positive samples showed the highest pH values. A higher decrease of the pH after freezing was seen in positive samples. Thus, the kind of conservation of milk samples should be considered, as well as the results of parameters that could be influenced by such variation. KEYWORDS: pH, freezing, milk, ewes.

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.

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

O presente trabalho 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.

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

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

This study analyzed the influence of the number of milkings, number of births, and udder quarter in immunoglobulin (Ig) concentration in the colostrum of healthy Holstein cows. It was collected two samples of colostrum by manual milking, getting the first jets to completion of bacteriological examination and immunoglobulin levels by radial immunodiffusion test in agar gel. Positive samples for bacteriological examination were excluded from this investigation. Medians of immunoglobulin's G, A and M in the colostrum collected before the first and second milking were respectively 9,200 and 6,400mg/dL (p=0.0029); 400 and 200mg/dL (p=0.0018); 800 and 400mg/dL (p=0.0001). Median immunoglobulin concentration in animals that calved once, twice or three times or in cows that calved 4 to 6 times were 6,400; 6,400; 3,200 and 11,200mg/dL IgG; 100, 200, 100 and 800mg/dL IgA ; and 400, 400, 100 and 800mg/dL IgM, respectively. Concentrations of IgG, IgA and IgM were greater in animals that calved more than 4 times (p0,05) entre os quartos mamários. Os teores de Igs do colostro de vacas Holandesas sofrem influência do número de ordenhas pós-parto e número de lactações. Estas variações podem ser consideradas fatores de risco associados à falha na transferência de imunidade passiva em bezerros neonatos, predispondo-os às doenças e ocasionando perdas produtivas á pecuária.

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

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

O objetivo deste trabalho foi avaliar o perfil microbiológico e celular do leite no período lactante e de involução ativa de ovelhas da raça Santa Inês. Foram avaliadas amostras lácteas de 12 ovelhas durante estes distintos períodos. Realizou-se o exame físico da mama, sendo as amostras lácteas submetidas à contagem de células somáticas (CCS), ao California Mastitis Test (CMT), ao exame microbiológico e aos testes de sensibilidade in vitro dos patógenos encontrados. Foram observados maiores escores do exame físico, CCS, CMT durante o período de involução ativa, além de uma alta persistência da infecção durante estes períodos. O período de involução ativa não se mostrou como um momento de alta susceptilidade. Os estafilococos coagulase negativa representaram o único gênero isolado das glândulas infectadas. Uma alta sensibilidade dos agentes etiológicos envolvidos frente aos diferentes antimicrobianos in vitro foi também observada. The aim was to evaluate and compare the microbiological and cellular profile of the milk of Santa Ines ewes during the lactation period and the active involution. Milk samples were analyzed from 12 ewes during these distinct periods. Clinical examination of the mammary gland, somatic cell count (SCC), California Mastitis Test (CMT), bacteriologic screening and sensibility of the pathogens in vitro were performed. Most alterations were observed in the active involution period. SCC and CMT were higher in this same period. Besides this, a high persistency of infection occurred. The active involution period did not show high susceptibility. Coagulase-negative staphylococci were the only isolated bacteria. A high antimicrobial sensibility of these pathogens was also encountered.

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.

A influencia do estagio de lactacao na composicao do leite de cabras foi estudada em 27 animais lactantes da raca Saanen durante oito meses, colhendo-se o total de 304 amostras. Foram selecionados e mantidos, no decorrer do trabalho, animais saudaveis, sem qualquer alteracao no exame fisico da glândula mamaria e cujo leite manteve-se negativo ao exame microbiologico. Avaliaram-se os teores de gordura, proteina, lactose e solidos totais. A concentracao de solidos totais, gordura e lactose declinaram durante a lactacao, porem os teores de proteina foram praticamente estaveis durante o periodo estudado. Os resultados obtidos permitem concluir que a constituicao do leite de cabras sofreu influencia do estagio de lactacao.

The hematological and immunological development of calves from birth to 6 months of age was performed by hemogram and cellular phenotype. Ten male Holstein calves were assessed in 13 moments: before colostrum intake (D0), every 2 days until the 10th day of life (D2 to D10), at the 15th day after birth (D15), and then monthly up to 6 months (D180). Calves presented hemoconcentration on day (D) 0 and showed a gradual decrease in hematimetric rates until D180. The inversion of the neutrophilic for lymphocytic profile was observed on D4. The percentage of CD3+ cells on D10 was higher than D30 up to D180. The number of CD4+ and CD8+ cells did not change between time points. The number of CD21+ lymphocytes was significantly higher at early time points of D0 up to D15, compared on D30 until D150. In conclusion, the neonatal period was marked by stress leukogram in the first 4 days, and low number of B lymphocytes. These might be risk factors for bacterial infections responsible for navel inflammation and diarrhea. The increase in the number of B cells from 30 days of age demonstrated that the calves were functional and able to generate an immune response. RESUMO O desenvolvimento hematológico e o imunológico de bezerros do nascimento aos seis meses de idade foram avaliados por meio hemograma e imunofenotipagem dos linfócitos. Para tanto, 10 bezerros machos Holandeses foram avaliados em 13 momentos: antes da ingestão do colostro (D0), a cada dois dias até o 10º dia (D2 a D10), 15º dia (D15) e, em seguida, mensalmente até seis meses de idade (D180). Os bezerros apresentaram hemoconcentração em D0 e decréscimo gradual dos índices hematimétricos até D180. A inversão do perfil neutrofílico para linfocítico foi observada em D4. Os valores de CD3+ (%) em D10 foram maiores que aqueles observados em D30 até D180. CD4+ e CD8+ não apresentaram alterações entre os momentos. Os linfócitos B CD21+ aumentaram significativamente a partir dos primeiros momentos D0 até D15, em comparação com os momentos subsequentes D30 até D150. Em conclusão, o período neonatal foi caracterizado pelo leucograma de estresse nos primeiros quatro dias e o baixo número de linfócitos B. Esses fatos podem ser considerados como fatores de risco para infecções bacterianas responsáveis pelas inflamações umbilicais e pela diarreia. Por outro lado, o aumento de células B após os 30 dias de vida demonstrou que os bezerros foram capazes de gerar uma resposta imune.