Butyrate supplementation in the liquid diet of dairy calves leads to a rapid recovery from diarrhea and reduces its occurrence and relapses in the preweaning period.

The present study aimed to evaluate the effect of continuous butyrate administration in dairy calves' liquid diet considering diarrhea, metabolic profile, gastrointestinal development, and corporal growth. Immediately after birth, calves were randomly allocated into 2 groups of 62 calves (50 females and 12 males), with access to water and a solid feed ad libitum. The butyrate group (BG) received 4 g/d of sodium butyrate (Admix Easy, Adisseo) diluted in the whole milk, and the control group (CG) received whole milk with no supplementation. Sodium butyrate was administered from d 1 of life until the weaning at 90 d. Feces consistency was assessed daily for the first 30 d of life and characterized by scores from 0 to 4 (0 and 1 for normal, and 2, 3, and 4 for abnormal feces). Diarrhea was diagnosed when the animals had abnormal feces and fever. Morbidity, recurrence, mortality, and lethality data were recorded and compared between the groups. Average daily gain (ADG) and corporal growth (body weight, thoracic perimeter, height at the withers, and croup width) were evaluated weekly, from the first day to d 30, and later at 45, 60, and 90 d of life. Blood samples were taken weekly for up to 30 d to determine the circulating levels of total calcium, phosphorus, chloride, bicarbonate, glucose, ß-hydroxybutyrate, and nonesterified fatty acids. The males were euthanized at 15 (n = 6 per group) and 30 d (n = 6 per group) for morphometric, histological, and gene expression analysis of the gastrointestinal tract. The results showed that the BG had a lower rate of morbidity (BG = 30% vs. CG = 50%) and recurrence (BG = 26.7% vs. CG = 60%) of diarrhea than the CG. In addition, the BG had abnormal feces for a shorter period (BG = 4.64 ± 0.47 d vs. CG = 8.6 ± 0.65 d). The ADG tended to be higher in BG than CG up to 30 and 60 d. Metabolic evaluations showed the lowest levels of glucose and highest levels of nonesterified fatty acids in BG. On d 30 of life, rumen papillae length, papilla area, duodenum villus length, and crypt depth were higher in BG than in CG. The duodenal gene expression at 30 d showed that animals with diarrhea episodes that did not receive butyrate had the highest levels of transcripts for the LCT and GLP2 genes. In addition, in different ways, both butyrate and neonatal diarrhea affected the gene expression of IGF1, SLC5A1, and AQP3. These results allow us to conclude that continuous supplementation with sodium butyrate improves gastrointestinal development, reduces the occurrence of diarrhea, and makes clinical conditions milder with faster recovery, favoring a higher ADG in the first 30 and 60 d of life. Based on these results, we conclude that sodium butyrate can be indicated for liquid diet supplementation to accelerate gastrointestinal tract development and prevent severe cases of neonatal diarrhea, tending to improve average daily gain until weaning.

Maternal supplementation with cobalt sources, folic acid, and rumen-protected methionine and its effects on molecular and functional correlates of the immune system in neonatal Holstein calves.

Calves born to multiparous Holstein cows fed during the last 30 d of pregnancy 2 different cobalt sources [cobalt glucoheptonate (CoPro) or cobalt pectin (CoPectin)], folic acid (FOA), and rumen-protected methionine (RPM) were used to study neonatal immune responses after ex vivo lipopolysaccharide (LPS) challenge. Groups were (n = 12 calves/group) CoPro, FOA+CoPro, FOA+CoPectin, and FOA+CoPectin+RPM. Calves were weighed at birth and blood collected at birth (before colostrum), 21 d of age, and 42 d of age (at weaning). Growth performance was recorded once a week during the first 6 wk of age. Energy metabolism, inflammation, and antioxidant status were assessed at birth through various plasma biomarkers. Whole blood was challenged with 3 µg/mL of LPS or used for phagocytosis and oxidative burst assays. Target genes evaluated by real-time quantitative PCR in whole blood samples were associated with immune response, antioxidant function, and 1-carbon metabolism. The response in mRNA abundance in LPS challenged versus nonchallenged samples was assessed via Δ = LPS challenged - LPS nonchallenged samples. Phagocytosis capacity and oxidative burst activity were measured in neutrophils and monocytes, with data reported as ratio (percentage) of CD14 to CH138A-positive cells. Data including all time points were subjected to ANOVA using PROC MIXED in SAS 9.4 (SAS Institute Inc.), with Treatment, Sex, Age, and Treatment × Age as fixed effects. A 1-way ANOVA was used to determine differences at birth, with Treatment and Sex as fixed effects. Calf birth body weight and other growth parameters did not differ between groups. At birth, plasma haptoglobin concentration was lower in FOA+CoPro compared with CoPro calves. We detected no effect for other plasma biomarkers or immune function due to maternal treatments at birth. Compared with CoPro, in response to LPS challenge, whole blood from FOA+CoPectin and FOA+CoPectin+RPM calves had greater mRNA abundance of intercellular adhesion molecule 1 (ICAM1). No effect for other genes was detectable. Regardless of maternal treatments, sex-specific responses were observed due to greater plasma concentrations of haptoglobin, paraoxonase, total reactive oxygen metabolites, nitrite, and ß-carotene in female versus male calves at birth. In contrast, whole blood from male calves had greater mRNA abundance of IRAK1, CADM1, and ITGAM in response to LPS challenge at birth. The longitudinal analysis of d 0, 21, and 42 data revealed greater bactericidal permeability-increasing protein (BPI) mRNA abundance in whole blood from FOA+CoPectin versus FOA+CoPro calves, coupled with greater abundance in FOA+CoPro compared with CoPro calves. Regardless of maternal treatments, most genes related to cytokines and cytokine receptors (IL1B, IL10, TNF, IRAK1, CXCR1), toll-like receptor pathway (TLR4, NFKB1), adhesion and migration (ICAM1, ITGAM), antimicrobial function (MPO), and antioxidant function (GPX1) were downregulated over time. Phagocytosis capacity and oxidative burst activity in both neutrophils and monocytes did not differ due to maternal treatment. Regardless of maternal treatments, we observed an increase in the percentage of neutrophils capable of phagocytosis and oxidative burst activity over time. Overall, these preliminary assessments suggested that maternal supplementation with FOA and Co combined with RPM had effects on a few plasma biomarkers of inflammation at birth and molecular responses associated with inflammatory mechanisms during the neonatal period.

The occurrence of diseases and their relationship with passive immune transfer in Holstein dairy calves submitted to individual management in southern Brazil / Ocorrência de doenças e sua relação com a transferência de imunidade passiva em bezerras Holandês criadas em sistema individual no sul do Brasil

Calves are extremely dependent on colostrum intake for the acquisition of passive immunity. This study aimed to determine the occurrence of diarrhea and respiratory diseases and the impact of Failure of Passive Immune Transfer (FPIT) on the health and zootechnical performance of Holstein dairy calves in individual management. This study has been carried out in five commercial farms in Rio Grande do Sul State, Brazil, from March 2017 to January 2018. In this study, 131 calves were followed from birth to 60 days of age. Total Plasmatic Protein (TPP) has been performed to determine passive immune transfer quality in 53 calves (53/131). A daily clinical follow-up has been accomplished aiming at diagnosing diseases and their incidences, and zootechnical measures such as withers height, width of the croup and weight have been evaluated. FPIT rate was 32.07%, diarrhea occurrence and respiratory diseases were 77.9% and 49.6%, respectively. FPIT increased the chances of calves presenting diarrhea and developing respiratory diseases, but no differences on zootechnical performance were found. The frequency of FPIT is still high and is a factor that corroborated the increased risk for diarrhea and respiratory disease but did not influence the performance of calves in the preweaning phase.(AU)

Bezerras são extremamente dependentes da ingestão de colostro para adquirir imunidade passiva. Este estudo teve o objetivo de determinar os índices de ocorrência de diarreia e de doença respiratória, assim como o impacto da falha na transferência de imunidade passiva (FTIP) no desenvolvimento de doenças e no desempenho zootécnico de bezerras Holandês criadas em sistema individual. O estudo foi desenvolvido em cinco propriedades comerciais no Rio Grande do Sul, Brasil, entre março de 2017 e janeiro de 2018. Assim, 131 bezerras foram acompanhadas, do nascimento aos 60 dias de idade. Em 53 animais, foi realizada avaliação de proteínas plasmáticas totais para determinar a qualidade na transferência de imunidade passiva. Acompanhamento clínico diário foi realizado, a fim de diagnosticar doenças e suas incidências, assim como avaliações zootécnicas, como altura de cernelha, largura de garupa, perímetro torácico e peso. O percentual de FTIP foi 32,07%, a ocorrência de diarreias e de doenças respiratórias foi, respectivamente, 77,9% e 49,6%. A FTIP aumentou as chances de as bezerras apresentarem diarreia e doenças respiratórias, mas não alterou o desempenho zootécnico. Conclui-se que a frequência na FTIP ainda é elevada, fator que corroborou o aumento do risco para as diarreias e doença respiratória. Apesar disso, a FTIP não influenciou no desenvolvimento das bezerras na fase de aleitamento.(AU)

Enrofloxacina de rápida ação associada ao tratamento com eletrólitos orais e reposição energética no tratamento de broncopneumonia neonatal em bezerros Holandeses / Fast action enrofloxacin associated with support treatment with oral electrolyte and energy replacement in the treatment of neonatal bronchopneumonia in Holstein calves

O objetivo deste estudo foi avaliar estratégias terapêuticas para o tratamento de infecções broncopulmonares, utilizando a enrofloxacina de ação rápida e sua associação com suporte e fluidoterapia endovenosa ou suporte e solução oral energética e eletrolítica, por meio da mensuração de parâmetros clínicos, hematológicos, bioquímicos e desenvolvimento corporal de neonatos bovinos. Foram utilizadas 35 bezerras da raça Holandesa, monitoradas desde o nascimento até a sexta semana de vida, divididas aleatoriamente nos grupos: grupo CONTROLE; grupo antibiótico; grupo antibiótico + suporte + fluidoterapia endovenosa; grupo antibiótico + suporte + solução oral; e grupo SUPORTE. Os parâmetros zootécnicos foram avaliados do nascimento até a sexta semana de vida, e os parâmetros hematológicos e bioquímicos foram avaliados zero, 24, 72 e 120 horas após diagnóstico da broncopneumonia. Os animais do grupo antibiótico + suporte + solução oral apresentaram menores níveis de eosinófilos e maiores níveis de neutrófilos segmentados em comparação aos animais dos demais grupos. Não houve diferença nos parâmetros zootécnicos avaliados. Neste estudo, o tratamento com antibiótico e solução oral ofereceu aos animais melhor aporte para combater a broncopneumonia, favorecendo o organismo a desenvolver uma resposta imune efetiva diante da infecção.(AU)

The objective of this study was to evaluate therapeutic strategies for treatment of bronchopulmonary infections using fast-acting enrofloxacin and its association with support and endovenous fluid or support and oral energy and electrolytic solution, by measuring clinical, hematological, biochemical and development parameters of bovine neonates. Thirty-five Holstein calves, monitored from birth to six weeks of age, were randomly divided into five groups: control group; antibiotic group; antibiotic group + support + intravenous fluid therapy; antibiotic group + support + oral solution; and support group. The performance parameters were evaluated from birth to the 6th week of age and hematological and biochemical parameters were evaluated 0, 24, 72 and 120 hours after diagnosis of bronchopneumonia. Calves of the antibiotic group + support + oral solution group presented lower levels of eosinophils and higher levels of segmented neutrophils compared to the other groups. There was no difference in performance parameters evaluated. In this study, the treatment with antibiotic and oral solution offered the animals had a better contribution to treat bronchopneumonia, favoring the organism to develop an effective immune response to that infection.(AU)

Limits of grape byproduct inclusion in diets for lambs: Zinc supplementation to prevent copper poisoning.

The aims of the current study were to assess the inclusion levels of grape byproduct replacing jiggs hay in lambs diets and to evaluate signs of copper poisoning. Thirty-four Texel and Corriedale crossbred female lambs were used in a randomized block experimental design. The treatments comprised four levels of grape byproduct (0; 100; 200 and 300 g/kg DM) in replacement of jiggs hay. The diets were adjusted to the same ZnCu ratio (6:1) of the basal diet. Grape byproduct consumption at up to 30%, led to similar weight gain in the different treatments (P = .92), which was suitable for growing lambs. Grape byproduct in the diet had linear effect on GGT (P < .001) and AST (P < .0001) enzymes as well as on total bilirubin (P = .05). In addition, the highest grape byproduct addition showed the highest consumption of hay (P < .01). Hay replacement by grape byproduct at up to 300 g/kg in the DM was satisfactory to weight gain and did not negatively affect feed intake and weight gain of growing lambs. Maintaining zinc:copper ratio in sheep diets is not effective in preventing liver damage caused by increased dietary copper concentrations over a period of 70 days.

Methionine and choline supply during the peripartal period alter polymorphonuclear leukocyte immune response and immunometabolic gene expression in Holstein cows.

Polymorphonuclear leukocytes (PMNL) are the first responders upon pathogen invasion and hence play an important role in inflammatory and immune responses. Rumen-protected methionine (MET) and choline (CHOL) during the peripartal period affect the immune response and inflammatory status in dairy cows to different extents. We aimed to examine the effect of MET and CHOL supply on expression of genes regulating key PMNL functions and associations with whole-blood immune challenge. Thirty multiparous Holstein cows from a larger cohort randomly assigned from -21 to 30 d relative to parturition to a basal control (CON) diet, CON plus MET at a rate of 0.08% of dry matter, or CON plus CHOL at 60 g/d were used. Blood was sampled at -10, 7, and 30 d relative to parturition for inflammatory biomarker analyses and PMNL isolation. Neutrophil and monocyte phagocytosis and oxidative burst in vitro were assessed in whole blood at 1, 7, and 28 d. Although neutrophil and monocyte phagocytosis did not differ, oxidative burst in neutrophils and monocytes was greater in MET-supplemented cows relative to CON cows. Compared with CON, PMNL adhesion and migration-related genes (ITGAM, ITGB2, ITGA4) were downregulated in response to MET and CHOL. Expression of CADM1 and SELL was also lower in MET-supplemented cows compared with CON cows but not in CHOL cows. In contrast, compared with CON cows, the expression of ICAM1 was lower in CHOL but not MET cows. Similar to adhesion and migration-related genes, cows receiving MET- or CHOL-supplemented diets had lower expression of inflammation-related genes (IL1ß, IL10RA, NFKB1, STAT3, TLR2). However, expression of IRAK1 and TLR4 was lower in MET- but not CHOL-supplemented cows. Plasma taurine concentration was greater in MET cows compared with CHOL and CON cows, suggesting a better redox status in plasma. In agreement with plasma taurine, oxidative stress-related genes (CBS, CTH, GPX1, GSS, SOD2) in PMNL were lower in response to MET and to CHOL supply. Overall, immunometabolic gene expression profile and blood biomarker analyses suggest an overall better redox status in PMNL during the transition period in response to MET and CHOL supply. These adaptations in PMNL might be beneficial for mounting a better bactericidal response upon challenge.

Effects of rumen-protected methionine and choline supplementation on steroidogenic potential of the first postpartum dominant follicle and expression of immune mediators in Holstein cows.

Multiparous Holstein cows were assigned in a randomized complete block design into four treatments from 21 d before calving to 30 d in milk (DIM). Treatments were: MET [n = 19, fed the basal diet + rumen-protected methionine at a rate of 0.08% (w/w) of the dry matter, Smartamine® M], CHO (n = 17, fed the basal diet + choline 60 g/d, Reashure®), MIX (n = 21, fed the basal diet + Smartamine® M at a rate of 0.08% (w/w) of the dry matter and 60 g/d Reashure®), and CON (n = 20, no supplementation, fed the close-up and fresh cow diets). Follicular development was monitored via ultrasound every 2 d starting at 7 DIM until ovulation (n = 37) or aspiration (n = 40) of the first postpartum dominant follicle (DF). Follicular fluid from 40 cows was aspirated and cells were retrieved immediately by centrifugation. Gene expression of TLR4, TNF, IL1-ß, IL8, IL6, LHCGR, STAR, 3ß-HSD, P450scc, CYP19A1, IRS1, IGF, MAT1A, and SAHH, was measured in the follicular cells of the first DF. Cows in CON had higher TNF, TLR4, and IL1-ß mRNA expression (11.70 ± 4.6, 21.29 ± 10.4, 6.28 ± 1.4, respectively) than CHO (2.77 ± 0.9, 2.16 ± 0.9, 2.29 ± 0.7, respectively), and MIX (2.23 ± 0.7, 1.46 ± 0.6, 2.92 ± 0.8, respectively). Cows in CON had higher IL1-ß expression (6.27 ± 1.4) than cows in MET (3.28 ± 0.6). Expression of IL8 mRNA was lower for cows in CHO (0.98 ± 0.3) than cows in CON (4.90 ± 0.7), MET (6.10 ± 1.7), or MIX (5.05 ± 1.8). Treatments did not affect mRNA expression of LHCGR, STAR, P450scc, CYP19A, SAHH, MAT1A, or IL6 however, 3ß-HSD expression was higher for cows in MET (1.46 ± 0.3) and MIX (1.25 ± 0.3) than CON (0.17 ± 0.04) and CHO (0.26 ± 0.1). Supplementation of methionine, choline, and both methionine and choline during the transition period did not affect days to first ovulation or number of cows that ovulated the first follicular wave. Plasma and follicular fluid estradiol and progesterone concentrations were not different among treatments. Methionine concentrations in the follicular fluid of the first postpartum DF was higher for cows in MET (18.2 ± 0.1 µM) than cows in CON (11.1 ± 0.9 µM). In conclusion, supplementing choline and methionine during the transition period changed mRNA expression in follicular cells and dietary methionine supplementation increased plasma and follicular fluid concentrations of methionine of the first postpartum DF in Holstein cows.

Combinação de butafosfan e cianocobalamina no metabolismo da glicose em vacas leiteiras após o parto / Combined of butaphosphan and cyanocobalamin on the glucose metabolism of dairy cows after calving

A hipótese deste estudo é de que o uso da combinação de butafosfan e cianocobalamina pode melhorar a resistência periférica à insulina, aumentar a quantidade de glicose disponível para a glândula mamária e a produção de leite. Assim, o objetivo foi investigar os efeitos combinados de butafosfan e cianocobalamina sobre o metabolismo da glicose em vacas leiteiras no período pós-parto. Vinte e uma vacas leiteiras foram divididas em dois grupos: grupo controle (CON, n= 11), que recebeu cinco aplicações de solução salina (20mL / animal 0,9% NaCl), e grupo Catosal(r) (ABC, n= 10), que recebeu cinco aplicações de 20mL de uma solução contendo as substâncias butafosfan e cianocobalamina (B12 Catosal(r), 100mg da substância butafosfan e 50µg de cianocobalamina por mL). As aplicações foram realizadas por via intramuscular, nos dias sete, 12, 17, 22 e 27 pós-parto. As amostras de sangue foram coletadas para a avaliação das concentrações plasmáticas de fósforo, glicose, ácidos graxos não esterificados (AGNE), albumina, aspartato aminotransferase (AST) e creatina quinase (CK). Nos dias oito e 28 pós-parto, os animais foram pesados e submetidos aos testes de tolerância à glicose e à insulina. O tratamento promoveu perda de peso (ABC 40,4kg, CON 10,73kg, P<0,05) e aumento da AST (ABC 62,92 ±3,31U/L, CON 53,11±3,49 U / L, P<0,05) e dos níveis de CK (ABC 134,09± 19,08U / L, CON 79,43 ± 18,27U / L). Os grupos não diferiram quanto ao metabolismo (área sob a curva) da glicose nos dias oito e 28, porém os animais tratados tiveram um aumento na glicemia (P<0,05) no dia 28 pós-parto (97,54 ± 8,54mg / dL), após a administração de insulina, em comparação ao dia oito (83,01 ± 8,54mg / dL). Assim, pode-se concluir que a combinação de butafosfan e cianocobalamina melhora a adaptação do metabolismo da glicose em vacas leiteiras no início da lactação.(AU)

The hypothesis of this study is that the combined use of butaphosphan and cyanocobalamin could enhance peripheral insulin resistance, increasing the amount of glucose available for the mammary gland and milk production. Thus, our aim was to investigate the combined effects of butaphosphan and cyanocobalamin on the glucose metabolism in dairy cows during the postpartum period. Twenty one dairy cows were divided into two groups: Control Group (CON, n= 11), that received 5 injections of saline solution (20mL/animal 0.9 % NaCl), and Catosal group (ABC, n= 10) which received 5 injections of 20mL of a Butafosfan and Cyanocobalamin solution (Catosal(r) B12, 100mg of Butafosfan and 50µg Cyanocobalamin for mL). The injections were performed by intramuscular route, on days 7, 12, 17, 22 and 27 postpartum. Blood samples were taken on these days to evaluate plasma concentrations of phosphorus, glucose, non-esterified fatty acids (NEFA), albumin, aspartate aminotransferase (AST) and creatine kinase (CK). On days 8 and 28 postpartum, the animals were weighted and subjected to the glucose tolerance and insulin challenge tests. The treatment promoted weight loss (ABC 40.4kg, CON 10.73kg, P< 0.05) and increased AST (ABC 62.92 ±3.31U/L, CON 53.11 ±3.49U/L, P< 0.05) and CK levels (ABC 134.09 ±19.08U/L, CON 79.43 ±18.27U/L). Glucose metabolism (area under the curve) did not differ (P> 0.05) among groups on days 8 and 28; however, ABC animals showed higher serum glucose levels (P< 0.05) after insulin administration on day 28 postpartum (97.54 ±8.54mg/dL) when compared to day 8 (83.01 ±8.54mg/dL). It could be concluded that the combined use of butaphosphan and cyanocobalamin interferes positively with the adaptation of glucose metabolism in dairy cows in early lactation.(AU)

Maternal supplementation with rumen-protected methionine increases prepartal plasma methionine concentration and alters hepatic mRNA abundance of 1-carbon, methionine, and transsulfuration pathways in neonatal Holstein calves.

An important mechanism of nutritional "programming" induced by supplementation with methyl donors during pregnancy is the alteration of mRNA abundance in the offspring. We investigated the effects of rumen-protected Met (RPM) on abundance of 17 genes in the 1-carbon, Met, and transsulfuration pathways in calf liver from cows fed the same basal diet without (control, CON) or with RPM at 0.08% of diet dry matter/d (MET) from -21 through +30 d around calving. Biopsies (n = 8 calves per diet) were harvested on d 4, 14, 28, and 50 of age. Cows fed RPM had greater plasma concentration of Met (17.8 vs. 28.2 µM) at -10 d from calving. However, no difference was present in colostrum yield and free AA concentrations. Greater abundance on d 4 and 14 of betaine-homocysteine S-methyltransferase 2 (BHMT2), adenosylhomocysteinase (AHCY; also known as SAHH), and cystathionine-ß-synthase (CBS) in MET calves indicated alterations in Met, choline, and homocysteine metabolism. Those data agree with the greater abundance of methionine adenosyltransferase 1A (MAT1A) in MET calves. Along with CBS, the greater abundance of glutamate-cysteine ligase (GCLC) and glutathione reductase (GSR) on d 4 in MET calves indicated a short-term postnatal alteration in the use of homocysteine for taurine and glutathione synthesis (both are potent intracellular antioxidants). The striking 7-fold upregulation at d 50 versus 4 of cysteine sulfinic acid decarboxylase (CSAD), catalyzing the last step of taurine synthesis, in MET and CON calves underscores an important role of taurine during postnatal calf growth. The unique role of taurine in the young calf is further supported by the upregulation of CBS, GCLC, and GSR at d 50 versus 14 and 28 in MET and CON. Although betaine-homocysteine S-methyltransferase (BHMT) activity did not differ in MET and CON, it increased ∼50% at d 14 and 28 versus 4. A significant positive correlation (r = 0.79) was present between BHMT abundance and BHMT activity regardless of treatment. The gradual upregulation over time of BHMT2 and SAHH coupled with the gradual upregulation of MAT1A and the DNA (cytosine-5-)-methyltransferases (DNMT1, DNMT3A, DNMT3B) in MET and CON calves was indicative of adaptations potentially driven by differences in intake of milk replacer and starter feed as calves grew. In that context, the ∼2.5-fold increase in abundance of DNMT3B at d 50 versus 4 in MET and CON indicate that DNA methylation might be an important component of the physiologic adaptations of calf liver. The data indicate that calves from MET-supplemented cows underwent alterations in Met, choline, and homocysteine metabolism partly to synthesize taurine and glutathione, which would be advantageous for controlling metabolic-related stress. Whether the effects in MET calves were directly related to increased Met supply in utero remains to be determined.

Effects of rumen-protected methionine and choline supplementation on the preimplantation embryo in Holstein cows.

Our objective was to determine the effects of supplementing methionine and choline during the prepartum and postpartum periods on preimplantation embryos of Holstein cows. Multiparous cows were assigned in a randomized complete-block design into four treatments from 21 days before calving to 30 days in milk (DIM). Treatments (TRT) were MET (n = 9, fed the basal diet + rumen-protected methionine at a rate of 0.08% [w:w] of the dry matter [DM], Smartamine M), CHO (n = 8, fed the basal diet + choline 60 g/d, Reashure), MIX (n = 11, fed the basal diet + Smartamine M and 60 g/d Reashure), and CON (n = 8, no supplementation, fed the close-up and fresh cow diets). Cows were randomly reassigned to two new groups (GRP) to receive the following diets from 31 to 72 DIM; control (CNT, n = 16, fed a basal diet) and SMT (n = 20, fed the basal diet + 0.08% [w:w] of the dry matter intake as methionine). An progesterone intravaginal insert (CIDR) device was inserted in all cows after follicular aspiration (60 DIM) and superovulation began at Day 61.5 using FSH in eight decreasing doses at 12-hour intervals over a 4-day period. On Days 63 and 64, all cows received two injections of PGF2α, and CIDR was removed on Day 65. Twenty-four hours after CIDR removal, ovulation was induced with GnRH. Cows received artificial insemination at 12 hours and 24 hours after GnRH. Embryos were flushed 6.5 days after artificial insemination. Global methylation of the embryos was assessed by immunofluorescent labeling of 5-methylcytosine, whereas lipid content was assessed by staining with Nile red. Nuclear staining was used to count the total number of cells per embryo. There was no difference between TRT, GRP, or their interaction (P > 0.05) for embryo recovery, embryos recovered, embryo quality, embryo stage, or cells per embryo. Methylation of the DNA had a TRT by GRP interaction (P = 0.01). Embryos from cows in CON-CNT had greater (P = 0.04) methylation (0.87 ± 0.09 arbitrary units [AU]) than embryos from cows in MET-CNT (0.44 ± 0.07 AU). The cytoplasmic lipid content was not affected (P > 0.05) by TRT or their interaction, but lipid content was greater (P = 0.04) for SMT (7.02 ± 1.03 AU) than that in CNT (3.61 ± 1.20 AU). In conclusion, cows in MET-CNT had embryos with lower methylation, and SMT cows had a higher lipid content than CNT. Methionine supplementation seems to impact the preimplantation embryo in a way that enhances its capacity for survival because there is strong evidence that endogenous lipid reserves serve as an energy substrate.

Effect of butaphosphan and cyanocobalamin on postpartum metabolism and milk production in dairy cows.

The aim of this study was to determine the effect of butaphosphan and cyanocobalamin (BTPC) supplementation on plasma metabolites and milk production in postpartum dairy cows. A total of fifty-two Holstein cows were randomly assigned to receive either: (1) 10 ml of saline (NaCl 0.9%, control group); (2) 1000 mg of butaphosphan and 0.5 mg of cyanocobalamin (BTPC1 group); and (3) 2000 mg of butaphosphan and 1.0 mg of cyanocobalamin (BTPC2 group). All cows received injections every 5 days from calving to 20 days in milk (DIM). Blood samples were collected every 15 days from calving until 75 DIM to determine serum concentration of glucose, non-esterified fatty acids (NEFA), ß-hydroxybutyrate (BHB), cholesterol, urea, calcium (Ca), phosphorus (P), magnesium (Mg), aminotransferase aspartate (AST) and γ-glutamyltransferase (GGT). The body condition score (BCS) and milk production were evaluated from calving until 90 DIM. Increasing doses of BTPC caused a linear reduction in plasma concentrations of NEFA and cholesterol. Supplementation of BTPC also reduced concentrations of BHB but it did not differ between the two treatment doses. Milk yield and milk protein had a linear increase with increasing doses of BTPC. A quadratic effect was detected for milk fat and total milk solids according to treatment dose, and BTPC1 had the lowest mean values. Concentrations of glucose, urea, P, Mg, AST, GGT, milk lactose and BCS were not affected by treatment. These results indicate that injections of BTPC during the early postpartum period can reduce NEFA and BHB concentrations and increase milk production in Holstein cows.

Resposta clínica e metabólica de potros neonatos em relação aos achados histopatológicos da placenta na égua / Clinical and metabolic response of neonatal foals related to histopathology finds in mare placenta

Avaliaram-se as respostas clínica e metabólica de potros neonatos em relação aos achados histopatológicos da placenta na égua. Foram avaliados dois grupos de éguas da raça Puro Sangue Inglês - um grupo-problema (n=25) e um grupo-controle (n=25), de acordo com os achados da placenta. O exame dos potros constou de avaliação clínica geral, hematologia e bioquímica sérica. O exame histopatológico da placenta apresentou resultado compatível com a apresentação clínica do potro, sendo que a presença de lesões inflamatórias resultou na produção de potros debilitados. A presença de lesões degenerativas não comprometeu o estado clínico do neonato, mas pode ser responsável pela manifestação de distúrbios subclínicos, evidenciados pelo aumento das taxas de AST e GGT. A ureia pareceu ser um indicador de dano renal decorrente de insuficiência placentária em potros neonatos.

The placenta represents the major communication between the mare and the fetus during the gestational period, and this suggests that any disturbance in the placenta can be an indicator of gestational damage with risk to the fetus. The aim of this paper was to evaluate the clinical and metabolic responses of the newborn foals related with the findings from the histopathological examination of the placenta. This study was conducted in a farm located in Bagé-RS, Brazil, where were evaluated two groups of Throughbred mares for this case-control study: One Problem Group (N=25) and the Control Group (n=25), based on the placental findings. The foal's evaluation was based on general clinical examination, hematology and serum biochemistry. Results from the placenta histopathological exams were compatible with clinical presentation of the foals, with the presence of inflammatory lesions resulting in the production of debilitated foals. The presence of degenerative lesions in the placenta does not compromise the clinical features of the newborn, but they can be responsible for the manifestation of sub-clinical disturbances, evidenced by increased levels of AST and GGT. Urea seems to be an indicator of renal damage due to placental insufficiency in neonatal foals.

Dietary lipid during the transition period to manipulate subcutaneous adipose tissue peroxisome proliferator-activated receptor-γ co-regulator and target gene expression.

Objectives were to determine adipose tissue mRNA expression of peroxisome proliferator-activated receptor (PPAR)γ co-regulators, target enzymes and transcription regulators, inflammation-related genes, and adipokines in response to dietary long-chain fatty acids (LCFA). From -21 through 10 d relative to parturition cows were fed no supplemental LCFA (control), saturated LCFA (SFAT; mainly 16:0 and 18:0), or fish oil (FO). Lipid was fed at 250 g/d prepartum or approximately 1.5 to 1.9% of the previous day's dry matter intake postpartum. Transcript profiling of 35 genes via quantitative PCR was conducted on biopsies (n=5 cows/diet) collected at -14 and 11 d from parturition. Despite lower dry matter intake with FO, pre- and postpartal blood nonesterified fatty acids, ß-hydroxybutyrate, and liver triacylglycerol were unaffected by treatment but increased after calving regardless of diet. Prepartal expression of adipogenic/lipogenic transcription regulators [CEBPA, CEBPB, RXRA, KLF5, and MLXIPL (formerly ChREBP)] and co-regulators (CARM1, EP300, NCOA1, MED1, NCOR2, and NRIP1) was upregulated by FO and SFAT versus control, whereas most enzymes involved in lipogenesis/triacylglycerol synthesis (FASN, SCD, DGAT2, and LPIN1) had greater expression only with FO. Expression of most adipogenic/lipogenic genes decreased after parturition, but feeding SFAT led to sustained upregulation of CEBPA, CEBPB, RXRA, several PPAR-co-activators, and DGAT2 and SCD, suggesting maintenance of a pro-adipogenic/pro-lipogenic state with SFAT. The co-activator CREBBP was greater in cows fed lipid and did not decrease after parturition, suggesting ligand activation of PPARγ. The greater peripartal expression of NFKB1 and TBK1 due to dietary lipid was suggestive of a local inflammatory response. At amounts fed prepartum, both FO and SFAT were effective in upregulating the adipose tissue PPARγ-gene network. In contrast, only SFAT led to sustaining that response. Overall, the observed expression patterns are suggestive of an adipogenic regulatory mechanism particularly responsive to SFAT.