Influence of bovine pestivirus heterogeneity on serological responses to 10 different commercial vaccine formulation.

Bovine Pestivirus typically involves one or more organ systems, with clinical manifestations ranging from mild to severe fatal systemic illness that lead to significant reproductive, productive, and economic losses. Vaccines face the challenge of addressing the significant variability of pestiviruses, which affects the interaction between viral antigens and the immune system's ability to provide protection. This study aimed to evaluate the serological responses against bovine viral diarrhea virus 1 (Pestivirus A) and Pestivirus B induced by 10 commercial vaccines, including one recombinant (vaccine E), two modified live (MLV multivalent, vaccine I, and MLV monovalent, vaccine J), and seven killed vaccines (KLV, vaccines A to H). Additionally, we evaluated the cross-reactivity between Pestivirus A and B from vaccines and HoBi-like pestivirus (Pestivirus H). In Phase 1, guinea pigs were used to screen for non-MLVs. They were divided into nine groups (n=6 each) and received two doses (⅕ of bovine dose) of eight different non-MLV on Days 0 and 21. Serum samples were collected on Days 0 and 30 for serological analyse. In Phase 2, Holstein × Gir heifers (n= 45) were divided into five groups, comprising 6-9 animals. They were vaccinated either once with MLVs or twice with the top non-MLVs screened in Phase 1. Serum samples were harvested on d0 (vaccination day) and d60 (60 days after the first dose) for MLV and non-MLV. Specific antibody titers were assessed virus neutralization (VN) and transformed in log2 for statistical analysis using PROC-MIXED. Significant effects were observed for vaccine groups, time points, and their interactions concerning neutralizing antibodies against Pestivirus A and B in both Guinea pigs and heifers. The Phase 1 study revealed serological responses against Pestivirus A exclusively in non-MLV D (85.33±13.49) and E (72.00±19.26). In the bovine study, the KLD vaccine D (72.00±15.10), recombinant vaccine E (90.66±25.85), and MLV I (170.66±28.22) resulted in an average of neutralizing antibodies against Pestivirus A that exceeded the protective threshold (≥ 60). However,individual analysis of heifers showed a higher frequency of animals presenting titers of Pestivirus A Ab surpassing 32 following vaccination with MLV I and J. None of the vaccine formulations in either study elicited a protective immune response against Pestivirus B or demonstrated cross-reactivity against Pestivirus H.

Serological Responses of Guinea Pigs and Heifers to Eight Different BoAHV-1 Vaccine Formulations.

Bovine alphaherpesvirus 1 (BoAHV-1) infection affects the production and reproductive performance of dairy and beef livestock, resulting in considerable economic losses. In addition to biosecurity measures, vaccination programs are effective strategies for controlling and preventing BoAHV-1 infection and transmission. We evaluated the serological immune response against BoAHV-1 induced by eight different formulations of commercial vaccines: three modified live vaccines and five killed vaccines containing BoAHV type 1 or types 1 and 5. In the first experiment, 50 BoAHV-1-seronegative guinea pigs were assigned to eight groups; each individual in the treatment groups received two doses (one-fifth of the bovine dose). The second experiment was conducted using 29 crossbred Holstein × Gir heifers in four groups of six to nine animals each. The serological immune response against BoAHV-1 was measured using virus neutralization and enzyme-linked immunosorbent assays to measure the total IgG against BoAHV. We evaluated the effects of the vaccine, time, and interaction of the vaccine and time on neutralizing antibodies against BoAHV-1. Killed vaccines produced low levels of antibodies against BoAHV-1, whereas modified live vaccines produced high levels of antibodies capable of providing neutralizing titers in the vaccinated animals, with the thermosensitive modified live vaccine showing the highest levels of antibodies.

Biosecurity practices in the dairy farms of southern Brazil.

Biosecurity refers to a set of practices that prevents and/or controls the introduction, spread, and elimination of harmful biological agents in a production system. In this study, we aimed to survey the biosecurity practices and determine their correlation with the size of production systems. A biosecurity assessment form was provided to 69 farms in the Campos Gerais region of Paraná, Brazil. The questionnaire was divided into two sections: general and bovine viral diarrhea virus- and bovine herpesvirus type-1-specific sections. The general section covered topics on traffic control, quarantine and animal isolation, hygiene practices, carcass disposal, and disease monitoring/control. The specific section consisted of questions on the reproductive and respiratory factors, use of antimicrobials, and vaccination schedule. The 69 farms were also classified into small (≤ 61), medium (62-201), and large (≥ 202) size farms based on the number of lactating cows. Moreover, multiple correspondence analysis (MCA) was performed between the biosecurity measures and farm size. The main risk factors and variability were related to the traffic control of people, animals, and vehicles/equipment, animal quarantine/isolation, and hygiene practices. MCA revealed that the small farms exhibited a lack of biosecurity measures, including those related to traffic control, animal quarantine, and hygiene. In medium-size farms, contact between bovine animals of different ages and difficulty in animal isolation in the quarantine system were among the main risk factors. In contrast, isolation of sick animals was easy, but the need to frequently purchase cattle was an important risk factor in large farms. These results highlight the relationship between biosecurity measures and farm size, providing valuable insights for the development of better biosecurity plans for production systems.

Effect of Prepartum Maternal Supplementation with Diphenyl Diselenide on Biochemical, Immunological, and Oxidative Parameters of the Offspring.

This study aimed to assess the impact of prepartum maternal diphenyl diselenide (PhSe)2 supplementation on the development, biochemical, immune, and antioxidant parameters of calves. Eighteen Holstein breed calves were used, born to females who were or were not subjected to supplementation, at 42, 28, and 14 days prior to calving. The (PhSe)2 group (DDG) was administered 3 µmol/kg of (PhSe)2 in 4 mL of dimethyl sulfoxide (DMSO), while the DMSO and NaCl groups were administered 4 mL of DMSO and 0.9% NaCl, subcutaneously. The calves were evaluated based on their weight, withers height, body condition score 24 h post-birth (0), as well on days 14, 28, 42, 56, 70. Blood samples were also taken to determine serum variables. Calves on the DDG showed higher average levels of total protein, albumin, and globulins on day 0, and the immunoglobulin G level was significantly higher than the other groups on days 0, 14, 56, 70. Maternal supplementation showed immunomodulatory effect on calves, evidenced by the exceptional rates of passive immunity transfer, as well as the enhancement of humoral immunity. Our research offers fresh insights into the immunomodulatory potential of (PhSe)2, making it a viable alternative in facing this challenging phase, rearing dairy calves.

Influence of Mycoplasma bovis infection on milk production and quality of Holstein dairy cows.

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).

Hematologic profiles and development of innate immune function in healthy Holstein calves during the pre-weaning period.

BACKGROUND: The development and state of innate immune cell function during the first 90 days of life in dairy calves have not been fully described. OBJECTIVE: 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. METHODS: 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. RESULTS: 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. CONCLUSIONS: 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.

Diagnostic accuracy of Wisconsin and California scoring systems to detect bovine respiratory disease in preweaning dairy calves under subtropical environmental conditions.

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.

Efficacy of prepartum vaccination against neonatal calf diarrhea in Nelore dams as a prevention measure.

BACKGROUND: Neonatal calf diarrhea (NCD) is the leading cause of calf morbidity and mortality in beef cattle. Cow's vaccination in last stage of pregnancy is one of the most important measures to mitigate the risk of NCD outbreaks. The aim of this study was to evaluate the efficacy of prepartum single dose vaccination against NCD, especially Bovine Rotavirus type A (BoRVA) and Bovine Coronavirus (BCoV), in Nelore dams and offspring. A total of 117 pregnant cows (n = 81) and heifers (n = 36) were distributed in two groups, vaccinated (VAC: cows = 40; heifers = 19) and non-vaccinated (NVAC: cows = 41; heifers = 17). Vaccination occurred between 60 to 50 days before the expected calving date with a single dose of a water-in-oil (W/O) vaccine, and NVAC group received a dose of saline solution 0.9%. Blood samples were collected before vaccination and 30 days after to evaluate the antibody (Ab) response. Specific IgG1 Abs against BoRVA and BCoV were measured by using an Enzyme Linked Immuno Sorbent Assay (ELISA). Calves' births were monitored, and the transference of passive immunity was evaluated. Diarrhea was monitored in the first 30 days of age, and fecal samples were collected for identification of the etiological agent. RESULTS: Higher titers of IgG1 Ab against BoRVA and BCoV was observed in the VAC group than NVAC group in the cow (P < 0.0001) and total dams categories (P < 0.0001). The titer of specific IgG1 Abs in the calves' serum reflected the dams response, observing higher IgG1 Ab titers for BoRVA (P < 0.0016) and BCoV (P < 0.0095) in the offspring born to VAC cows and higher IgG1 Ab titers for BoRVA(P < 0.0171) and BCoV (P < 0.0200) in the offspring born to VAC total dams. The general incidence of diarrhea observed was 18.6% (11/59) and 29.3% (17/58) in the calves born to the VAC and NVAC group, respectively. CONCLUSIONS: Prepartum vaccination with a single dose of the vaccine tested increased the titers of IgG1 Ab against BCoV and BoRVA, and it could be used as a preventive strategy to decrease the NCD occurrence in Nelore calves.

Effect of selenium and vitamin E supplementation on the metabolic status of dairy goats and respective goat kids in the peripartum period.

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.

Effect of prophylactic use of tulathromycin on gut bacterial populations, inflammatory profile and diarrhea in newborn Holstein calves.

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.

Somatic cell count in buffalo milk using fuzzy clustering and image processing techniques.

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.

Relationships among Indicators of Metabolism, Mammary Health and the Microbiomes of Periparturient Holstein Cows.

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.

Influence of feeding fresh colostrum from the dam or frozen colostrum from a pool on indicator gut microbes and the inflammatory response in neonatal calves.

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.

An Assessment of Secondary Clinical Disease, Milk Production and Quality, and the Impact on Reproduction in Holstein Heifers and Cows from a Single Large Commercial Herd Persistently Infected with Bovine Viral Diarrhea Virus Type 2.

The aim of this study was to evaluate secondary clinical disease, milk production efficiency and reproductive performance of heifers and cows persistently infected (PI) with bovine viral diarrhea virus type 2 (BVDV type 2). PI animals (n = 25) were identified using an antigen capture ELISA of ear notch samples. They were distributed into three age groups: ≤ 12 (n = 8), 13 to 24 (n = 6) and 25 to 34 (n = 11) months old. A control group of BVDV antigen ELISA negative female cattle that were age matched to the PI animals was utilized from the same herd. The PI group had a 1.29 higher odds ratio for diarrhea than controls (p = 0.001, IC95% = 1.032-1.623) and 1.615 greater chance of developing bovine respiratory disease (BRD) (p = 0.012, IC95% = 1.155-2.259). The age at first insemination (p = 0.012) and number of insemination attempts required to establish the first pregnancy (p = 0.016) were both higher for PI than controls. Milk production was higher for control cows than PI cows during most of the sampling periods. Somatic cell counts (SCC) were higher in PI cows than the controls at all sampling points across lactation (p ≤ 0.042). PI cattle had a higher incidence of disease, produced less milk, a higher SCC, and poorer reproductive performance than control cattle in this study.

Influence of early use of antimicrobial on the health and performance of Holstein calves in the first month of life / Influência do uso precoce de antimicrobiano na sanidade de bezerras holandesas no primeiro mês de vida

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.(AU)

O uso precoce de antimicrobianos tem sido adotado em muitas fazendas para profilaxia das diarreias e doença respiratória em bezerras, no entanto existem controvérsias sobre os beneficios desta prática na saúde desses animais. Esta pesquisa avaliou a influência do uso precoce de antimicrobiano na sanidade e desempenho de bezerras holandesas recém-nascidas. Para tanto foram selecionadas 26 bezerras Holandesas distribuídas de acordo com a aplicação (ATB+) ou não (ATB-) de tulatromicina (2,5mg/Kg) por via subcutânea até 12h de vida. As bezerras foram examinadas por meio de exame clínico geral, escore fecal, escore respiratório e palpação externa da região umbilical, além da matéria seca fecal. A presença de anemias foi determinada pelo eritrograma utilizando sistema automático e além da dosagem de ferro utilizando kit comercial. O diagnóstico etiológico das diarreias foi investigado por meio da técnica de flutuação em solução saturada de sacarose (Cryptosporidium) e multiplex semi-nested RT-PCR (rotavírus/coronavírus). O desempenho das bezerras foi estimado pelo ganho de peso. As bezerras foram avaliadas até doze horas após o nascimento (≤12h); 3-5º (D3-5); 7-9º (D7-9); 13-15º (D13-15); 20-23º (D20-23); e 27-30º dias de vida (D27-30). Não foram encontradas diferenças entre os grupos ATB+ e ATB- em relação à frequência cardíaca, frequência respiratória e temperatura retal. O eritrograma revelou maior frequência de anemias no grupo ATB- (P=0,016) no D3-5. Neste momento também foram observados menores valores de ferro sérico (P=0,051). Foram detectados treze casos de doença respiratória durante o estudo, no entanto não foi possível detectar diferença entre os grupos. A frequência de diarreias (escore fecal 2 e 3) foi alta em ambos os grupos, observando-se pico no D13-15 (ATB+=92,3%; ATB-=92,3%). Não observamos diferenças entre os grupos em relação a frequência de diarreia considerando-se a matéria seca fecal. O agente etiológico predominante nas diarreias foi o Cryptosporidium. O ganho de peso diário foi igual entre grupos, com intensa redução no GPD no D13-15. A administração de tulatromicina na dose profilática (2,5mg/Kg) ao nascimento diminuiu a frequência de anemias e não influenciou no ganho de peso e prevalência de diarreias.(AU)

Effect of the type of milking on mammary gland examination in Saanen goats / Efeito do tipo de ordenha sobre o exame da glândula mamária de cabras Saanen

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.(AU)

Esta pesquisa avaliou o efeito da ordenha sobre o exame físico da glândula mamária (GM) e do leite em cabras Saanen. Seis propriedades foram selecionadas, quatro fazendas foram utilizadas após avaliação da seleção dos critérios e distribuídas em dois grupos de acordo com o método de ordenha: manual ou mecânico. O exame físico da GM foi realizado utilizando as técnicas de inspeção e palpação, além do exame do leite pelo teste da caneca de fundo escuro, California Mastitis Test, contagem de células somáticas e exame bacteriológico. Foi possível detectar uma associação entre o método de ordenha com a maioria dos exames físicos da GM: inspeção da GM (p = 0,001), inspeção do teto (p = 0,002), palpação da GM (p = 0,054) e palpação do teto (p = 0,036). A ordenha mecânica associou-se com a redução do volume de GM (OR = 6,00), redução do tamanho do teto (OR = 16,19) e consistência firme do parênquima mamário (OR = 2,39). O uso da ordenha mecânica representou menor risco para o aumento do volume do volume da GM (OR = 0,288) e presença da cisternite (OR = 0,490). Também foi detectada a associação entre a CCS e o tipo de ordenha (p = 0,002). A ordenha mecânica foi associada com as alterações no exame físico de GM e do leite compatíveis com lesões traumáticas. Essas informações indicam o uso inadequado do equipamento de ordenha na criação de cabras leiteiras.(AU)

Serological response against bovine herpesvirus and bovine viral diarrhea virus induced by commercial vaccines in Holstein heifers / Resposta sorológica contra herpesvírus bovino e vírus da diarreia viral bovina induzida por vacinas comerciais em novilhas Holandesas

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.(AU)

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.(AU)

Effects of 3 Different Commercial Vaccines Formulations against BVDV and BHV-1 on the Inflammatory Response of Holstein Heifers.

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 48h (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.

Specific immune response in neonate Holstein heifer calves fed fresh or frozen colostrum / Resposta imune específica em bezerras holandesas recém-nascidas alimentadas com colostro fresco ou congelado

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.(AU)

O objetivo desta pesquisa foi avaliar a influência das células do colostro no desenvolvimento imune em bezerras leiteiras durante os primeiros 28 dias de vida. Os animais foram distribuídas em 2 grupos: COL+ (n=9) recebeu colostro fresco de suas próprias mães; e COL- (n=10) recebeu pool de colostro congelado sem células viáveis oriundo de vacas doadoras. Estas bezerras foram avaliadas antes da ingestão do colostro (D0); às 48 horas (D2) e semanalmente entre o D7 e D28. O desenvolvimento da imunidade foi avaliada pela fenotipagem das subpopulações celulares do sangue e produção de citocinas pelas células mononucleares sanguíneas após 72 horas de estimulação com concanavalina A (ConA), Staphylococcus aureus (S. aureus) e Escherichia coli (E. coli) inativadas. O histórico clínico das bezerras foi marcado por elevada frequência de diarreias em ambos os grupos. Entretanto, COL- apresentou maior intensidade de diarreia (escore de fezes ~ 3 de 4) e maior temperatura retal no D7 do que as bezerras COL+. Além disso, broncopneumonia (n=1) e inflamações umbilicais foram diagnosticadas apenas nas bezerras COL-. O grupo COL- apresentou menor concentração de ferro sérico e maior número de linfócitos no D7 do que o grupo COL+. COL- também apresentou maior frequência de anemias que o grupo COL+ no D21 e D28. Em geral, a fração das subpopulações celulares foram semelhantes entre os dois grupos ao longo do tempo estudado, exceto na semana 1, onde as bezerras COL- apresentaram maior proporção de CD45RO+ (P=0.07). Observou-se um constante aumento de CD45RO+ com declínio concomitante de CD45RO- ao longo do estudo, indicando o desenvolvimento da resposta imune. A proporção de células CD14MHCII+ aumentou de acordo com a idade (P≤0.05). As medianas das citocinas produzidas a partir do PBMC não estimuladas apresentaram valores abaixo do nível de detecção em ambos os grupos. O PBMC do COL+ estimulado com ConA secretou elevada concentração de IL17 na semana 2 (COL+=2060.0pg/mL e COL-=0.0pg/ml, P=0.00). PBMC do COL+ estimuladas com S. aureus inativado (P=0.05) e E. coli inativada (P=0.05) secretaram níveis mais elevados de IL17 que as bezerras COL- na semana 4. Outros picos de IL17 foram observados no COL+ na semana 2, porém não foi possível detectar diferenças estatísticas entre os grupos. Conclui-se que as bezerras alimentadas com colostro fresco e congelado apresentaram perfil similar entre as proporções das subpopulações celulares. O aumento de células expressando os marcadores de memória CD45RO+ e CD14MHCII+ demonstram, ao longo do experimento, o amadurecimento do sistema imune específico das bezerras após estimulação natural por patógenos após o nascimento. A maior secreção de IL17 pelas células das bezerras COL+ indica que as células maternas podem modular resposta imune Th17 direcionada aos antígenos bacterianos. Este mecanismo poderia ser responsável pela rápida e eficiente quimiotaxia de neutrófilos e eliminação dos microrganismos bacterianos. Os diferentes perfis de citocinas podem ser responsáveis pelos diferentes históricos clínicos relatados para as bezerras COL+ e COL-.(AU)

Effect of maternal cells transferred with colostrum on the health of neonate calves.

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±1.4g/dL) than COL+ (RBC=7.2±0.8×106/µL; Hgb=9.6±1.3g/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±2.7µM/L) than COL+ (10.7±6.2µM/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.