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Adequate prediction of postruminal outflows of essential AA (EAA) is the starting point of balancing rations for EAA in dairy cows. The objective of this meta-analysis was to compare the performance of 3 dairy feed evaluation systems (National Research Council [NRC], Cornell Net Protein and Carbohydrate System version 6.5.5 [CNCPS], and National Academies of Sciences, Engineering and Medicine [NASEM]) to predict EAA outflows (Trp was not tested). The data set included a total of 354 treatment means from 70 duodenal and 24 omasal studies. To avoid Type I error, mean and linear biases were considered of concern if statistically significant and representing >5.0% of the observed mean. Analyses were conducted on raw observed values and on observations adjusted for the random effect of study. The analysis on raw data indicates the ability of the feed evaluation system to predict absolute values whereas the analysis on adjusted values indicates its ability to predict responses of EAA outflows to dietary changes. For the prediction of absolute values (based on raw data), NRC underpredicted outflows of all EAA, from 5.3% to 8.6% of the observed mean (%obs.mean) except for Leu, Lys, and Val; NASEM overpredicted Lys (10.8%obs.mean); and CNCPS overpredicted Arg, His, Lys, Met, and Val (5.2 to 26.0%obs.mean). No EAA had a linear bias of concern with NASEM, followed by NRC for His (6.8%obs.mean), and CNCPS for all EAA (5.6 to 12.2%obs.mean) except Leu, Phe, and Thr. In contrast, for the prediction of responses to dietary changes (based on adjusted data), NRC had 2 EAA presenting a linear bias of concern, followed by NASEM and CNCPS with 4 and 6 EAA, respectively. Predictions of His showed a linear bias of concern (5.3 to 9.6%obs.mean) with the 3 feed evaluation systems. Measured chemistry of crude protein and EAA were reported for 1 or more feed ingredients of the ration in 36% of the studies, and resulted in decreased linear biases in the 3 feed evaluation systems. The difference in mean biases of Met outflows was systematically positive when comparing omasal versus duodenal studies. Predictions of Met outflows with NRC had a higher concordance correlation coefficient in duodenal (used to develop NRC equations) versus omasal studies, whereas the opposite was observed with CNCPS, the latter showing the lowest mean bias for Met in omasal sampling studies. The 30% difference in Met mean biases between sampling sites appeared related to a similar difference found for observed Met versus nonammonia nitrogen outflows between duodenal and omasal studies, which is independent of predictions. In conclusion, NRC and NASEM yielded accurate predictions of EAA outflows, with a small superiority of NASEM to predict absolute values, and slight superiority of NRC to predict the responses to dietary changes. In comparison, CNCPS may present mean and linear biases of concern for many EAA. Moreover, it remains to determine which sampling site is more representative of the true supply of EAA to the cows.
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Aminoácidos , Alimentación Animal , Dieta , Bovinos , Animales , Aminoácidos/metabolismo , Femenino , Dieta/veterinaria , Rumen/metabolismo , DuodenoRESUMEN
Dairy cattle excreta are a valuable source of orthophosphate (Ortho-P), an inorganic form of phosphorus (P) that is readily available for microorganisms, plant growth, and development. There is, however, a growing environmental concern about the potential negative environmental impact of excessive amounts of Ortho-P excretion, which can lead to the eutrophication of water bodies. As a result, the development of mathematical equations to quantify and manage Ortho-P excretion on dairy farms could prove valuable for environmental sustainability. This study aimed to use literature data to develop empirical predictions for Ortho-P (g/kg dry matter [DM]) excretion using total P (TP [g/kg DM]) content of dairy cattle feces (Ortho-Pf) and manure (Ortho-Pm). Data sets from studies that evaluated and characterized the different forms of P in feces and manure from dairy cattle were compiled. After outlier exclusion, the final retained database for feces included 37 treatment means from 4 published papers while the manure comprised 23 treatment means from 7 published papers. A linear-mixed model was used to develop the predictive equations, incorporating the random effect of the study. A leave-one-out cross-validation procedure was used to evaluate the predictive ability of the developed models, whereby studies were regarded as folds. The fecal equation was determined as Ortho-Pf (g/kg DM) = -2.447 (0.572) + 0.966 (0.083) × TP (g/kg DM) (R2 = 0.79) and resulted in a root mean square prediction error as a percentage of mean observed value (RMSPE, %) of 32.8% and error due to random sources of 97.6%. Additionally, the manure equation was determined as Ortho-Pm (g/kg) = -0.204 (0.446) + 0.590 (0.065) × TP (g/kg) (R2 = 0.77) and had an RMSPE of 43.3% with a random error of 93.9%. Both models revealed minimal mean and slope biases on feces and manure data. Findings suggest that these sets of equations can be used to estimate excreted Ortho-P from total excreted P, helping nutritionists and farmers to understand the impact of dietary P changes on the environment. Further, these equations can be incorporated into extant models such as the Cornell Net Carbohydrate and Protein System (CNCPS) to aid in understanding and mitigating P and Ortho-P excretion from dairy cattle and to clarify the portion of P that migrates more rapidly into watersheds.
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Adequate prediction of postruminal outflow of protein fractions is the starting point for the determination of metabolizable protein supply in dairy cows. The objective of this meta-analysis was to compare the performance of 3 dairy feed evaluation systems (National Research Council [NRC], Cornell Net Protein and Carbohydrate System [CNCPS], and National Academies of Sciences, Engineering and Medicine [NASEM]) to predict outflows (g/d) of nonammonia nitrogren (NAN), microbial N (MiN), and nonammonia nonmicrobial N (NANMN). Predictions of rumen degradabilities (% of nutrient) of protein (RDP), NDF, and starch were also evaluated. The data set included 1,294 treatment means from 312 digesta flow studies. The 3 feed evaluation systems were compared using the concordance correlation coefficient (CCC), the ratio of root mean square prediction error (RMSPE) on standard deviation of observed values (RSR), and the slope between observed and predicted values. Mean and linear biases were deemed biologically relevant and are discussed if higher than a threshold of 5% of the mean of observed values. The comparisons were done on observed values adjusted or not for the study effect; the adjustment had a small effect on the mean bias but the linear bias reflected a response to a dietary change rather than absolute predictions. For the absolute predictions of NAN and MiN, CNCPS had the best-fit statistics (8% greater CCC; 6% lower RMSPE) without any bias; NRC and NASEM underpredicted NAN and MiN, and NASEM had an additional linear bias indicating that the underprediction of MiN increased at increased predictions. For NANMN, fit statistics were similar among the 3 feed evaluation systems with no mean bias; however, the linear bias with NRC and CNCPS indicated underprediction at low predictions and overprediction at elevated predictions. On average, the CCC were smaller and RSR ratios were greater for MiN versus NAN indicating increased prediction errors for MiN. For NAN responses to a dietary change, CNCPS also had the best predictions, although the mean bias with NASEM was not biologically relevant and the 3 feed evaluation systems did not present a linear bias. However, CNCPS, but not the 2 other feed evaluation systems, presented a linear bias for MiN, with responses being overpredicted at increased predictions. For NANMN, responses were overpredicted at increased predictions for the 3 feed evaluation systems, but to a lesser extent with NASEM. The site of sampling had an effect on the mean bias of MiN and NANMN in the 3 feed evaluation systems. The mean bias of MiN was higher in omasal than duodenal studies in the 3 feed evaluation systems (from 55 to 61 g/d) and this mean bias was twice as large when 15N labeling was used as a microbial marker compared with purines. Such a difference was not observed for duodenal studies. The reasons underlying these systematic differences are not clear as the type of measurements used in the current meta-analysis does not allow to delineate if one site or one microbial marker is yielding the "true" postruminal N outflows. Rumen degradabilities of protein was underpredicted with CNCPS, and RDP responses to a dietary change was underpredicted by the 3 feed evaluation systems with increased RDP predictions. Rumen degradability of NDF was underpredicted and had poor fit statistics for NASEM compared with CNCPS. Fit statistics were similar between CNCPS and NASEM for rumen degradability of starch, but with an underprediction of the response with NASEM and absolute values being overpredicted with CNCPS. Multivariate regression analyses showed that diet characteristics were correlated with prediction errors of N outflows in each feed evaluation system. Globally, compared with NAN and NANMN, residuals of MiN were correlated with several moderators in the 3 feed evaluation systems reflecting the complexity to measure and model this outflow. In addition, residuals of NANMN were correlated positively with RDP suggesting an overestimation of this parameter. In conclusion, although progress is still to be made to improve equations predicting postruminal N outflows, the current feed evaluation systems provide sufficient precision and accuracy to predict postruminal outflows of N fractions.
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Alimentación Animal , Compuestos de Nitrógeno , Femenino , Bovinos , Animales , Compuestos de Nitrógeno/metabolismo , Alimentación Animal/análisis , Dieta/veterinaria , Fibras de la Dieta/metabolismo , Almidón/metabolismo , Rumen/metabolismo , Lactancia/metabolismo , Proteínas en la Dieta/metabolismo , DigestiónRESUMEN
Greenhouse gas emissions, such as enteric methane (CH4) from ruminant livestock, have been linked to global warming. Thus, easily applicable CH4 management strategies, including the inclusion of dietary additives, should be in place. The objectives of the current study were to: (i) compile a database of animal records that supplemented monensin and investigate the effect of monensin on CH4 emissions; (ii) identify the principal dietary, animal, and lactation performance input variables that predict enteric CH4 production (g/d) and yield (g/kg of dry matter intake DMI); (iii) develop empirical models that predict CH4 production and yield in dairy cattle; and (iv) evaluate the newly developed models and published models in the literature. A significant reduction in CH4 production and yield of 5.4% and 4.0%, respectively, was found with a monensin supplementation of ≤24 mg/kg DM. However, no robust models were developed from the monensin database because of inadequate observations under the current paper's inclusion/exclusion criteria. Thus, further long-term in vivo studies of monensin supplementation at ≤24 mg/kg DMI in dairy cattle on CH4 emissions specifically beyond 21 days of feeding are reported to ensure the monensin effects on the enteric CH4 are needed. In order to explore CH4 predictions independent of monensin, additional studies were added to the database. Subsequently, dairy cattle CH4 production prediction models were developed using a database generated from 18 in vivo studies, which included 61 treatment means from the combined data of lactating and non-lactating cows (COM) with a subset of 48 treatment means for lactating cows (LAC database). A leave-one-out cross-validation of the derived models showed that a DMI-only predictor model had a similar root mean square prediction error as a percentage of the mean observed value (RMSPE, %) on the COM and LAC database of 14.7 and 14.1%, respectively, and it was the key predictor of CH4 production. All databases observed an improvement in prediction abilities in CH4 production with DMI in the models along with dietary forage proportion inclusion and the quadratic term of dietary forage proportion. For the COM database, the CH4 yield was best predicted by the dietary forage proportion only, while the LAC database was for dietary forage proportion, milk fat, and protein yields. The best newly developed models showed improved predictions of CH4 emission compared to other published equations. Our results indicate that the inclusion of dietary composition along with DMI can provide an improved CH4 production prediction in dairy cattle.
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Hypocalcemia induced by immune activation is a conserved response among mammals. Early postpartum cows will experience decreased circulating Ca concentrations following acute immune activation; however, the cause for decreased Ca concentration is unknown. Our objectives were to (1) describe Ca dynamics following an intravenous (IV) LPS challenge in early postpartum cows, and (2) compare inflammatory-induced changes in Ca dynamics between IV Ca-treated cows and control cows. Cows (n = 14, 8 ± 1 d in milk) were enrolled in a matched-pair randomized controlled design to receive IV Ca (IVCa) in a eucalcemic clamp for 12 h, or 0.9% NaCl (CTRL) following an IV LPS infusion (0.040 or 0.045 µg of LPS/kg of body weight over 1 h). During the 24 h following LPS infusion, circulating concentrations of parathyroid hormone and serotonin were measured, serum and urine samples were collected to calculate urinary fractional excretion of Ca (FECa), and fecal samples were collected to calculate Ca apparent digestibility (ADCa) using amylase-treated and ash-corrected undigested neutral detergent fiber after 240 h (uNDFom240) as an internal marker. Changes in Ca intake and milk Ca secretion were also quantified and compared with baseline values. Cows were fasted during challenge and dry matter intake was 20 ± 5% less than baseline values on the day of challenge and did not differ between groups. On the day of challenge, milk Ca concentration increased, but milk yield decreased such that total Ca secreted in milk did not change from baseline. Urine FECa was low overall, but an interaction of treatment and time was identified such that FECa increased in IVCa but decreased in CTRL. Concentrations of parathyroid hormone increased and serotonin decreased following challenge. Fecal dry matter decreased from baseline, but did not differ between 6, 12, and 24 h, and did not differ between groups. An interaction of treatment and time was identified for ADCa and apparent digestibility of dry matter such that digestibility was decreased in CTRL but not IVCa at 6 h. Acute immune activation induced hypocalcemia in CTRL, and although urinary Ca excretion was not a primary cause, it is unclear to what degree hypocalcemia was due to altered ADCa. Eucalcemia appeared to alter adaptations in Ca homeostasis during immune activation as FECa was increased in IVCa animals.
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Enfermedades de los Bovinos , Hipocalcemia , Femenino , Bovinos , Animales , Calcio , Lipopolisacáridos/efectos adversos , Hipocalcemia/veterinaria , Serotonina , Periodo Posparto , Lactancia/fisiología , Leche , Calcio de la Dieta , Hormona Paratiroidea , Dieta/veterinaria , Mamíferos , Enfermedades de los Bovinos/inducido químicamenteRESUMEN
Improving the ability of diet formulation models to more accurately predict AA supply while appropriately describing requirements for lactating dairy cattle provides an opportunity to improve animal productivity, reduce feed costs, and reduce N intake. The goal of this study was to evaluate the sensitivity of a new version of the Cornell Net Carbohydrate and Protein System (CNCPS) to formulate diets for rumen N, Met, and all essential AA (EAA). Sixty-four high-producing dairy cattle were randomly assigned to 1 of the 4 following diets in a 14-wk longitudinal study: (1) limited metabolizable protein (MP), Met, and rumen N (Base), (2) adequate Met but limited MP and rumen N (Base + M), (3) adequate Met and rumen N, but limited MP (Base + MU), and (4) adequate MP, rumen N, and balanced for all EAA (Positive). All diets were balanced to exceed requirements for ME relative to maintenance and production, assuming a nonpregnant, 650-kg animal producing 40 kg of milk at 3.05% true protein and 4.0% fat. Dietary MP was 97.2, 97.5, 102.3, and 114.1 g/kg of dry matter intake for the Base, Base + M, Base + MU, and Positive diets, respectively. Differences were observed for dry matter intake and milk yield (24.1 to 24.7 and 39.4 to 41.1 kg/d, among treatments). Energy corrected milk, fat, and true protein yield were greater (2.9, 0.13, and 0.08 kg/d, respectively) in cows fed the Positive compared with the Base diet. Using the updated CNCPS, cattle fed the Base, Base + M, and Base + MU diets were predicted to have a negative MP balance (-231, -310, and -142 g/d, respectively), whereas cattle fed the Positive diet consumed 33 g of MP/d excess to ME supply. Bacterial growth was predicted to be depressed by 16 and 17% relative to adequate N supply for the Base and Base + M diets, respectively, which corresponded with the measured lower apparent total-tract NDF degradation. The study demonstrates that improvements in lactation performances can be achieved when rumen N and Met are properly supplied and further improved when EAA supply are balanced relative to requirements. Formulation using the revised CNCPS provided predictions for these diets, which were sensitive to changes in rumen N, Met, all EAA, and by extension MP supply.
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Aminoácidos Esenciales , Metionina , Femenino , Bovinos , Animales , Metionina/metabolismo , Aminoácidos Esenciales/metabolismo , Lactancia , Suplementos Dietéticos , Rumen/metabolismo , Nitrógeno/metabolismo , Estudios Longitudinales , Proteínas de la Leche/análisis , Leche/química , Dieta/veterinaria , Racemetionina/metabolismo , Proteínas en la Dieta/metabolismoRESUMEN
Mycobacteria mediate horizontal gene transfer (HGT) by a process called distributive conjugal transfer (DCT) that is mechanistically distinct from oriT-mediated plasmid transfer. The transfer of multiple, independent donor chromosome segments generates transconjugants with genomes that are mosaic blends of their parents. Previously, we had characterized contact-dependent conjugation between two independent isolates of Mycobacterium smegmatis. Here, we expand our analyses to include five independent isolates of M. smegmatis and establish that DCT is both active and prevalent among natural isolates of M. smegmatis. Two of these five strains were recipients but exhibited distinct conjugal compatibilities with donor strains, suggesting an ability to distinguish between potential donor partners. We determined that a single gene, Msmeg0070, was responsible for conferring mating compatibility using a combination of comparative DNA sequence analysis, bacterial genome-wide association studies (GWAS), and targeted mutagenesis. Msmeg0070 maps within the esx1 secretion locus, and we establish that it confers mycobacterial self-identity with parallels to kin recognition. Similar to other kin model systems, orthologs of Msmeg0070 are highly polymorphic. The identification of a kin recognition system in M. smegmatis reinforces the concept that communication between cells is an important checkpoint prior to DCT commitment and implies that there are likely to be other, unanticipated forms of social behaviors in mycobacteria. IMPORTANCE Conjugation, unlike other forms of HGT, requires direct interaction between two viable bacteria, which must be capable of distinguishing between mating types to allow successful DNA transfer from donor to recipient. We show that the conjugal compatibility of Mycobacterium smegmatis isolates is determined by a single, polymorphic gene located within the conserved esx1 secretion locus. This gene confers self-identity; the expression of identical Msmeg0070 proteins in both donor-recipient partners prevents DNA transfer. The presence of this polymorphic locus in many environmental mycobacteria suggests that kin identification is important in promoting beneficial gene flow between nonkin mycobacteria. Cell-cell communication, mediated by kin recognition and ESX secretion, is a key checkpoint in mycobacterial conjugation and likely plays a more global role in mycobacterial biology.
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Mycobacterium smegmatis , Mycobacterium , Conjugación Genética , ADN/metabolismo , Estudio de Asociación del Genoma Completo , Mycobacterium/genética , Mycobacterium smegmatis/genética , Mycobacterium smegmatis/metabolismoRESUMEN
Functional characterization of bacterial proteins lags far behind the identification of new protein families. This is especially true for bacterial species that are more difficult to grow and genetically manipulate than model systems such as Escherichia coli and Bacillus subtilis To facilitate functional characterization of mycobacterial proteins, we have established a Mycobacterial Systems Resource (MSR) using the model organism Mycobacterium smegmatis This resource focuses specifically on 1,153 highly conserved core genes that are common to many mycobacterial species, including Mycobacterium tuberculosis, in order to provide the most relevant information and resources for the mycobacterial research community. The MSR includes both biological and bioinformatic resources. The biological resource includes (i) an expression plasmid library of 1,116 genes fused to a fluorescent protein for determining protein localization; (ii) a library of 569 precise deletions of nonessential genes; and (iii) a set of 843 CRISPR-interference (CRISPRi) plasmids specifically targeted to silence expression of essential core genes and genes for which a precise deletion was not obtained. The bioinformatic resource includes information about individual genes and a detailed assessment of protein localization. We anticipate that integration of these initial functional analyses and the availability of the biological resource will facilitate studies of these core proteins in many Mycobacterium species, including the less experimentally tractable pathogens M. abscessus, M. avium, M. kansasii, M. leprae, M. marinum, M. tuberculosis, and M. ulceransIMPORTANCE Diseases caused by mycobacterial species result in millions of deaths per year globally, and present a substantial health and economic burden, especially in immunocompromised patients. Difficulties inherent in working with mycobacterial pathogens have hampered the development and application of high-throughput genetics that can inform genome annotations and subsequent functional assays. To facilitate mycobacterial research, we have created a biological and bioinformatic resource (https://msrdb.org/) using Mycobacterium smegmatis as a model organism. The resource focuses specifically on 1,153 proteins that are highly conserved across the mycobacterial genus and, therefore, likely perform conserved mycobacterial core functions. Thus, functional insights from the MSR will apply to all mycobacterial species. We believe that the availability of this mycobacterial systems resource will accelerate research throughout the mycobacterial research community.
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Genes Bacterianos , Mycobacterium smegmatis/genética , Mycobacterium/genética , Investigación , Biología Computacional , Biblioteca de Genes , Mycobacterium/clasificación , Mycobacterium/patogenicidad , Mycobacterium smegmatis/crecimiento & desarrolloRESUMEN
The objective of this study was to evaluate the effect of rolled barley grain (RB) supplementation on rumen metabolism, omasal flow of nutrients, and microbial dynamics in lactating dairy cows fed fresh perennial ryegrass (Lolium perenne L.; PRG)-based diets. Ten ruminally cannulated Holstein cows averaging (mean ± standard deviation) 49 ± 23 d in milk and 513 ± 36 kg of body weight were assigned to 1 of 2 treatments in a switchback design. The treatment diets were PRG only (G) or PRG plus 3.5 kg of dry matter RB (G+RB). The study consisted of three 29-d periods where each period consisted of 21 d of diet adaptation and 8 d of data and sample collection. A double marker system was used to quantify nutrient flow entering the omasal canal along with labeled 15N-ammonium sulfate to measure bacterial, protozoal, and nonmicrobial N flow. Rumen evacuation techniques were used to determine nutrient and microbial pool size, allowing the calculation of fractional rates of digestion and microbial growth. There was no difference in daily milk yield or energy-corrected milk yield between treatments. Milk fat concentration and milk urea N decreased, whereas milk protein concentration increased in cows fed the G+RB diet. During the omasal sampling phase, dry matter intake was higher in cows fed the G+RB diet. Ruminal and total-tract neutral detergent fiber digestibility was lower in G+RB cows; however, no difference was observed in reticulorumen pH. The rumen pool size of fermentable carbohydrate was increased in cows fed the G+RB diet; however, the fractional rate of digestion was decreased. Flow of nonammonia N and bacterial N at the omasal canal increased in cows fed the G+RB diet compared with the G diet. Protozoa N flow was not different between diets; however, protozoa appeared to supply a much larger amount of microbial N and exhibited shorter generation time than previously considered. Feed N ruminal digestibility, corrected for microbial contribution, was similar for both treatments (88.4 and 89.0% for G and G+RB, respectively). In conclusion, RB supplementation did not benefit overall animal performance; however, it reduced ruminal neutral detergent fiber digestibility and increased bacterial N flow. The results demonstrate the large dependence of cows consuming PRG-based diets on microbial N as the main source of nonammonia N supply. Additional quantitative research is required to further describe the supply of nutrients and microbial dynamics in cows consuming PRG-based diets in an effort to determine most limiting nutrients.
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Bovinos/fisiología , Suplementos Dietéticos/análisis , Hordeum , Lolium , Leche/metabolismo , Animales , Peso Corporal , Bovinos/microbiología , Dieta/veterinaria , Fibras de la Dieta/metabolismo , Digestión , Grano Comestible , Femenino , Fermentación , Lactancia , Leche/química , Nutrientes/metabolismo , Omaso/metabolismo , Rumen/metabolismo , Rumen/microbiología , Urea/metabolismoRESUMEN
Strategies that can improve health and maximize growth in the preweaning period should improve the subsequent production and longevity of replacement animals. Few data are available that quantify feed and water consumption, as well as growth, in healthy versus non-healthy calves-the objective of this study. A database of Holstein calves (<1 wk of age; n = 313) was developed to compare calves that developed diarrhea in the first 21 d after arrival from commercial farms to the research facility versus calves that remained healthy. Individual calf data from 4 experiments included daily intake of milk replacer, free water, electrolyte solution, and starter grain, as well as weekly body weight (BW) and frame measures for 21 d after arrival. Calves with a fecal score of >2 for ≥3 consecutive days over the first 21 d of each experiment were retrospectively classified as diarrheic (DIA; n = 96); the remainder were classified as healthy (HEA; n = 217). Other health issues were minimal. The likelihood of elevated fecal score occurrence and the cumulative number of days with an elevated score were greater for DIA calves than for HEA calves. The initial total protein concentration in blood did not differ between classifications. Cumulative milk replacer dry matter intake (DMI) and water consumed from milk replacer were significantly less for DIA calves than for HEA calves, because DIA calves were more likely to refuse milk replacer. Cumulative starter DMI was decreased for DIA versus HEA calves. As a result, cumulative total DMI was significantly less for DIA calves than for HEA calves. Cumulative free water intake did not differ between classifications. The DIA calves were more likely to receive electrolyte solution and have more days given electrolyte solution than HEA calves. As a result, total cumulative intake of electrolyte solution was greater in DIA calves than in HEA calves. Cumulative total water intake did not differ between classifications. Initial BW did not differ between classifications; however, a classification × time interaction for BW indicated that HEA calves were heavier than DIA calves and had greater ADG. Significant classification × time interactions for hip height and heart girth revealed that HEA calves had a larger frame size. Gain-feed ratios for both milk replacer intake and total DMI differed between classifications: DIA calves were less feed-efficient than HEA calves. In conclusion, diarrhea in young calves decreases DMI, BW gain, and feed efficiency relative to HEA calves within 21 d of arrival.
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Enfermedades de los Bovinos/diagnóstico , Bovinos/crecimiento & desarrollo , Industria Lechera/métodos , Diarrea/veterinaria , Transportes , Alimentación Animal , Animales , Peso Corporal , Diarrea/diagnóstico , Dieta/veterinaria , Heces , Femenino , Masculino , Leche , Estudios Retrospectivos , Factores de TiempoRESUMEN
Autoimmune hepatitis (AIH) is a chronic liver disease characterized by progressive inflammation, female preponderance and seropositivity for autoantibodies such as anti-smooth muscle actin and/or anti-nuclear, anti-liver kidney microsomal type 1 (anti-LKM1) and anti-liver cytosol type 1 (anti-LC1) in more than 80% of cases. AIH is linked strongly to several major histocompatibility complex (MHC) alleles, including human leucocyte antigen (HLA)-DR3, -DR7 and -DR13. HLA-DR4 has the second strongest association with adult AIH, after HLA-DR3. We investigated the role of HLA-DR4 in the development of AIH by immunization of HLA-DR4 (DR4) transgenic non-obese diabetic (NOD) mice with DNA coding for human CYP2D6/FTCD fusion autoantigen. Immunization of DR4 mice leads to sustained mild liver injury, as assessed biochemically by elevated alanine aminotransferase, histologically by interface hepatitis, plasma cell infiltration and mild fibrosis and immunologically by the development of anti-LKM1/anti-LC1 antibodies. In addition, livers from DR4 mice had fewer regulatory T cells (Tregs ), which had decreased programmed death (PD)-1 expression. Splenic Tregs from these mice also showed impaired inhibitory capacity. Furthermore, DR4 expression enhanced the activation status of CD8+ T cells, macrophages and dendritic cells in naive DR4 mice compared to naive wild-type (WT) NOD mice. Our results demonstrate that HLA-DR4 is a susceptibility factor for the development of AIH. Impaired suppressive function of Tregs and reduced PD-1 expression may result in spontaneous activation of key immune cell subsets, such as antigen-presenting cells and CD8+ T effectors, facilitating the induction of AIH and persistent liver damage.
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Antígeno HLA-DR4/genética , Antígeno HLA-DR4/inmunología , Hepatitis Autoinmune/etiología , Hepatitis Autoinmune/patología , Amoníaco-Liasas , Animales , Células Presentadoras de Antígenos/inmunología , Células Presentadoras de Antígenos/metabolismo , Autoanticuerpos/inmunología , Autoantígenos/inmunología , Linfocitos T CD8-positivos/inmunología , Linfocitos T CD8-positivos/metabolismo , Citocromo P-450 CYP2D6/genética , Citocromo P-450 CYP2D6/inmunología , Citocinas/metabolismo , Modelos Animales de Enfermedad , Glutamato Formimidoiltransferasa , Humanos , Hipergammaglobulinemia/inmunología , Inmunización , Inmunoglobulina G/inmunología , Mediadores de Inflamación/metabolismo , Ratones , Ratones Endogámicos NOD , Ratones Transgénicos , Complejos Multienzimáticos/genética , Complejos Multienzimáticos/inmunología , Enzimas Multifuncionales , Células Plasmáticas/inmunología , Células Plasmáticas/metabolismo , Subgrupos de Linfocitos T/inmunología , Subgrupos de Linfocitos T/metabolismoRESUMEN
A new species, Porphyromonas gulae sp. nov., is proposed to include strains isolated from the gingival sulcus of various animal hosts which are distinct from related strains of Porphyromonas gingivalis of human origin. This bacterium exhibits the following characteristics: black-pigmented colonies; asaccharolytic, obligate anaerobic growth; and Gram-negative, non-motile and non-spore-forming, rod-shaped cells. Colonies do not fluoresce under UV light. Vitamin K1 and haemin are required for growth. Cells haemagglutinate sheep erythrocytes. Major fatty acid end products are butyric acid, isovaleric acid, succinic acid and phenylacetic acid. Strains are catalase-positive and indole is produced. Alkaline phosphatase, trypsin-like and N-acetyl-beta-glucosaminidase activities are strong. A beta-galactosidase and a glutamylglutamic acid arylamidase are also present. The G+C content of the chromosomal DNA is 51 mol%. DNA-DNA homology data and 16S rRNA gene sequence analysis provide strong evidence that strains from the animal biotype of P. gingivalis represent a Porphyromonas species that is distinct from P. gingivalis. The type strain of P. gulae is Loup 1T (= ATCC 51700T = NCTC 13180T).
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Encía/microbiología , Filogenia , Porphyromonas/clasificación , Animales , Secuencia de Bases , Carnívoros , Gatos , ADN Bacteriano/genética , Perros , Geografía , Haplorrinos , Humanos , Datos de Secuencia Molecular , Hibridación de Ácido Nucleico , Porphyromonas/genética , Porphyromonas/aislamiento & purificación , Porphyromonas gingivalis/clasificación , Porphyromonas gingivalis/genética , Técnica del ADN Polimorfo Amplificado Aleatorio , Homología de Secuencia de Ácido Nucleico , UrsidaeRESUMEN
OBJECTIVE: To study effects of in vitro exposure of bovine leukocytes to mercury, cadmium, and lead on phagocytosis, natural killer cell activity, and lymphocyte proliferation. SAMPLE POPULATION: Leukocytes from 6 nonpregnant Holstein heifers. PROCEDURE: Leukocytes were exposed in vitro to the aforementioned metals, and leukocyte functions were assessed. RESULTS: Phagocytosis was suppressed by 10(-5) to 10(-7) M CdCl2 and by 10(-5) and 10(-6) M HgCl2, but not 10(-7) M HgCl2 nor 10(-4) to 10(-6) M PbCl2. Spontaneous and concanavalin A- or phytohemagglutinin-stimulated proliferation of metal-treated bovine blood mononuclear cells was not significantly different from that of nontreated control cells, except for enhanced spontaneous proliferation in response to 10(-5) M HgCl2. When proliferation was expressed as a stimulation index, a dose-dependent increase of spontaneous proliferation was observed in response to exposure to HgCl2 and PbCl2. Compared with response to 10(-6) or 10(-7) M CdCl2, reduction of mitogen-induced and spontaneous proliferation was observed on exposure to 10(-5) M CdCl2. Natural killer cell activity against YAC-1 target cells, evaluated by flow cytometry, was decreased only in cells exposed to 10 M HgCl2. CONCLUSION AND CLINICAL RELEVANCE: Bovine leukocytes are susceptible to the immunomodulatory effects of in vitro exposure to heavy metals at concentrations equal to or higher than those at which similar effects are seen for leukocytes from most other animal species for which data are available for comparison. Exception is phagocytosis, which is severely affected by low concentrations of CdCl2 and HgCl2 in cattle. Reduction of defense mechanisms on exposure to metals could lead to increased susceptibility to potential pathogens.
Asunto(s)
Bovinos/inmunología , Leucocitos Mononucleares/inmunología , Metales Pesados/toxicidad , Animales , Cadmio/toxicidad , Células Cultivadas , Femenino , Citometría de Flujo/veterinaria , Células Asesinas Naturales/efectos de los fármacos , Células Asesinas Naturales/inmunología , Plomo/toxicidad , Leucocitos Mononucleares/efectos de los fármacos , Activación de Linfocitos/efectos de los fármacos , Mercurio/toxicidad , Microscopía Fluorescente/veterinaria , Fagocitosis/efectos de los fármacos , Conteo por Cintilación/veterinaria , Timidina/químicaRESUMEN
In order to assess the immunotoxic potential of food naturally contaminated with PCBs and other organohalogens, Fisher rats were fed a diet in which the lipids originated from the blubber of either a highly polluted St. Lawrence beluga or a relatively uncontaminated Arctic beluga. After a period of 2 months, different immune functions were evaluated, including lymphoblastic transformation, natural killer cell activity, plaque-forming cells, phagocytosis, oxidative burst, and immunophenotyping. For all assays, rats fed a St. Lawrence beluga blubber diet or a mixture of Arctic and St. Lawrence beluga blubber diet were not different from control rats fed a diet containing Arctic beluga blubber. These results are inconsistent with the well-known immunosuppressive effects of organochlorines in numerous species and with the lesions suggestive of organochlorine-related immunosuppression that are observed in St. Lawrence belugas. The lack of observable immunotoxic effects in rats fed contaminated beluga blubber might be explained by antagonistic effects in the organohalogen mixture, by a response specific to the rat, by a strain-related lack of sensitivity to organochlorines, or by insufficient dose due to the shortness of the exposure period or the route of exposure.
Asunto(s)
Tejido Adiposo/química , Grasas de la Dieta , Hidrocarburos Clorados , Sistema Inmunológico/efectos de los fármacos , Insecticidas/toxicidad , Contaminantes Químicos del Agua/toxicidad , Ballenas , Animales , Contaminación de Alimentos , Terapia de Inmunosupresión , Quebec , Ratas , Ratas Endogámicas F344RESUMEN
BACKGROUND & AIMS: Anti-liver cytosol type 1 autoantibodies have been reported in association with anti-liver-kidney microsome type 1 autoantibodies in 30% of patients with autoimmune hepatitis type II. In 10% of cases, anti-liver cytosol type 1 antibodies are the only liver-related circulating autoantibodies. The liver cytosol antigen is a liver-specific 62-kilodalton protein present in the cell as an oligomer of approximately 240 kilodaltons. The aim of this study was to identify the antigen recognized by anti-liver cytosol antibody. METHODS: To identify the liver cytosol antigen, an anti-liver cytosol type 1-positive serum was used for the screening of a complementary DNA library from HepG2 cells. Double immunodiffusion method was used to show the identity between the cytosolic and the cloned protein. RESULTS: The sequence of two isolated clones showed 85.2% homology with the formiminotransferase cyclodeaminase (FTCD) enzyme from pig liver. Antibodies purified by affinity with the recombinant protein and sera from mice immunized with FTCD recognized a 62-kilodalton human cytosolic protein when tested by immunoblot. The identity of precipitation lines was found between the cytosolic antigen and FTCD. CONCLUSIONS: This enzyme is a liver-specific antigen recognized by the sera of patients with autoimmune hepatitis.
Asunto(s)
Amoníaco-Liasas/inmunología , Autoanticuerpos/sangre , Autoantígenos/inmunología , Hepatitis Autoinmune/sangre , Hígado/enzimología , Secuencia de Aminoácidos , Amoníaco-Liasas/química , Amoníaco-Liasas/genética , Animales , Anticuerpos Monoclonales , Autoantígenos/química , Autoantígenos/genética , Secuencia de Bases , Carcinoma Hepatocelular , Clonación Molecular , Citosol/enzimología , Citosol/inmunología , Femenino , Biblioteca de Genes , Glutamato Formimidoiltransferasa , Hepatitis Autoinmune/inmunología , Humanos , Inmunodifusión , Hígado/inmunología , Neoplasias Hepáticas , Ratones , Ratones Endogámicos C57BL , Datos de Secuencia Molecular , Complejos Multienzimáticos , Enzimas Multifuncionales , Proteínas Recombinantes/química , Proteínas Recombinantes/inmunología , Alineación de Secuencia , Homología de Secuencia de Aminoácido , Homología de Secuencia de Ácido Nucleico , Porcinos , Células Tumorales CultivadasRESUMEN
Hormone-sensitive lipase (Lipe) catalyzes both the release lease of fatty acids from storage triglycerides in adipocytes and the liberation of cholesterol from cholesterol esters in steroidogenic tissues. Lipe activity is regulated in a tissue-, development- and hormone-specific fashion, the latter in large part by serine phosphorylation. We cloned and sequenced the Lipe gene from the 129Sv strain mouse, including 2.7 kb of the 5' nontranslated region. The primary transcript of the 129Sv Lipe locus spans 9.6 kb and contains 9 exons. We studied the curious hypervariable region immediately 5' to the regulatory serine residues by aligning the peptide and nucleic acid sequences of mouse, human, and rat Lipe. We propose that much of the variability is attributable to differences in the copy number of a 12-nucleotide repeat that shifts the intron 7 acceptor splice site. Introns 1 and 7 contain B1 elements, which in intron 7 are immediately adjacent to a tetranucleotide repeat. The mouse Lipe promoter region contains numerous potential binding motifs for factors implicated in adipose tissue expression and hormone responsiveness including adipocyte determination- and differentiation-dependent factor 1 (ADD1/SREBP1).
Asunto(s)
Hormonas/farmacología , Análisis de Secuencia de ADN , Esterol Esterasa/genética , Secuencia de Aminoácidos , Animales , Secuencia de Bases , Clonación Molecular , ADN Complementario , Humanos , Intrones , Mamíferos , Ratones , Datos de Secuencia Molecular , Ratas , Secuencias Repetitivas de Ácidos Nucleicos , Esterol Esterasa/químicaRESUMEN
Ketone bodies are produced in the liver, mainly from the oxidation of fatty acids, and are exported to peripheral tissues for use as an energy source. They are particularly important for the brain, which has no other substantial non-glucose-derived energy source. The 2 main ketone bodies are 3-hydroxybutyrate (3HB) and acetoacetate (AcAc). Biochemically, abnormalities of ketone body metabolism can present in 3 fashions: ketosis, hypoketotic hypoglycemia, and abnormalities of the 3HB/AcAc ratio. Normally, the presence of ketosis implies 2 things: that lipid energy metabolism has been activated and that the entire pathway of lipid degradation is intact. In rare patients, ketosis reflects an inability to utilize ketone bodies. Ketosis is normal during fasting, after prolonged exercise, and when a high-fat diet is consumed. During the neonatal period, infancy and pregnancy, times at which lipid energy metabolism is particularly active, ketosis develops readily. Pathologic causes of ketosis include diabetes, ketotic hypoglycemia of childhood, corticosteroid or growth hormone deficiency, intoxication with alcohol or salicylates, and several inborn errors of metabolism. The absence of ketosis in a patient with hypoglycemia is abnormal and suggests the diagnosis of either hyperinsulinism or an inborn error of fat energy metabolism. An abnormal elevation of the 3HB/AcAc ratio usually implies a non-oxidized state of the hepatocyte mitochondrial matrix resulting from hypoxia-ischemia or other causes. We summarize the differential diagnosis of abnormalities of ketone body metabolism, as well as pertinent recent advances in research.