Effects of altering diet carbohydrate profile and physical form on zoo-housed giraffe Giraffa camelopardalis reticulata.

Unlike wild giraffe that primarily consume low starch browse, the preference of zoo-housed giraffe for consuming supplemental feeds over forage could increase the risk of digestive disorders such as ruminal acidosis. Our objective was to evaluate the effects of modifying a supplemental feed's non-fibre carbohydrate profile and physical form on nutritional, behavioural, and blood measures of giraffe in a zoological setting. Six non-lactating, adult, female reticulated giraffes were used in a two-pen modified reversal study using two dietary treatments in seven 21-day periods with data collected on days 15-21. Dietary treatments were a control feed comprised of commercially available products used at the time as the giraffe feed (GF) and an unpelleted experimental feed (EF). On a dry matter basis, GF and EF, respectively, contained 17.0% and 17.4% crude protein, 14.2% and 1.5% starch, 14.9% and 21.3% ethanol-soluble carbohydrates, 22.9% and 26.0% acid detergent fibre (ADF) and 9.50% and 14.9% ND-soluble fibre (NDSF), with modulus of fineness values of 3.62 and 4.82. Supplemental feeds, alfalfa hay, salt, and water were available for ad libitum consumption. Significance was declared at p ≤ 0.05. Intakes of hay, supplemental feeds, and total feed did not differ by diet (p > 0.28), though intakes of starch (0.93 and 0.12 kg; p = 0.05) and ADF (1.83 and 2.23 kg; p = 0.04) differed between GF and EF respectively. Giraffe behaviour values (min/48 h) were greater with EF for total eating (p = 0.04); diets were not detected as different for engagement in oral stereotypes (GF = 433, EF = 318 min/48 h; p = 0.22). Blood glucose was higher on GF than EF (99.0 and 82.3 mg/dL; p = 0.03). The lower EF blood glucose value is more similar to ranges reported for domesticated ruminants. No differences were detected for changes in body weight or body condition score in the 21-day periods (p > 0.32). Modification of supplemental feed carbohydrate profile and physical form can influence behaviour and blood glucose values of zoo-housed giraffe.

Comparison of alternative neutral detergent fiber methods to the AOAC definitive method.

Neutral detergent fiber (NDF) is the most commonly reported metric for fiber in dairy cattle nutrition. An empirical method, NDF is defined by the procedure used to measure it. The current definitive method for NDF treated with amylase (aNDF) is AOAC Official Method 2002.04 performed on dried samples ground through the 1-mm screen of a cutting mill with refluxing and then filtration through Gooch crucibles without (AOAC-; reference method) or with (AOAC+) a glass fiber filter filtration aid. Other methods in use include grinding materials through the 1-mm screen of an abrasion mill, using filtration through a Buchner funnel with a glass fiber filter (Buch), and use of the ANKOM system (ANKOM Technology, Macedon, NY) that simultaneously extracts and filters samples through filter bags with larger (F57) or smaller (F58) particle size retentions. Our objective was to compare the AOAC and alternative methods using samples ground through the 1-mm screens of cutting or abrasion mills. Materials analyzed were 2 alfalfa silages, 2 corn silages, dry ground and high-moisture corn grains, mixed grass hay, ryegrass silage, soybean hulls, calf starter, and sugar beet pulp. Samples were run in duplicate in replicate analytical runs performed on different days by experienced technicians. Compared with cutting mill-ground samples, the aNDF% of dry matter results from abrasion mill-ground samples were or tended to be lower for 8 of 11 samples. Method affected aNDF% results for all materials, with method × grind interactions for 6 of 11 samples. For ash-free aNDF% assessed with cutting mill-ground materials, a priori selected contrasts showed that the number of materials for which methods differed or tended to differ from the AOAC methods were 4 (Buch), 8 (F57), and 3 (F58); and 3 for AOAC- versus AOAC+. However, statistically different does not necessarily mean substantially different. For a given feed and grind, a positive value for the absolute difference between the AOAC- mean and an alternative method mean minus 2 times the standard deviation of AOAC- suggests that values for the alternative method fall outside of the range of results likely to be observed for the reference method. The number of observed positive values for materials processed with cutting and abrasion mills, respectively, were 0 and 2 (AOAC+); 2 and 2 (Buch); 8 and 10 (F57); 4 and 7 (F58); and 0 and 4 (AOAC-). With the materials tested, methods in order of agreement with the reference method were Buch, F58, and F57, which often gave lower values. The AOAC+ gave results similar to AOAC-, substantiating it as an allowed modification of AOAC-. Best agreement between the reference method and variant NDF methods was achieved with the 1-mm screen cutting mill grind. The 1-mm abrasion mill grind produced more aNDF% results that were lower than the reference method but with fewer differences when filter particle retention size was smaller. The use of filters that retain finer particles could be explored to improve comparability of variant NDF methods and grinds. Further evaluation with an expanded set of materials is warranted.

Nutritional and greenhouse gas impacts of removing animals from US agriculture.

As a major contributor to agricultural greenhouse gas (GHG) emissions, it has been suggested that reducing animal agriculture or consumption of animal-derived foods may reduce GHGs and enhance food security. Because the total removal of animals provides the extreme boundary to potential mitigation options and requires the fewest assumptions to model, the yearly nutritional and GHG impacts of eliminating animals from US agriculture were quantified. Animal-derived foods currently provide energy (24% of total), protein (48%), essential fatty acids (23-100%), and essential amino acids (34-67%) available for human consumption in the United States. The US livestock industry employs 1.6 × 106 people and accounts for $31.8 billion in exports. Livestock recycle more than 43.2 × 109 kg of human-inedible food and fiber processing byproducts, converting them into human-edible food, pet food, industrial products, and 4 × 109 kg of N fertilizer. Although modeled plants-only agriculture produced 23% more food, it met fewer of the US population's requirements for essential nutrients. When nutritional adequacy was evaluated by using least-cost diets produced from foods available, more nutrient deficiencies, a greater excess of energy, and a need to consume a greater amount of food solids were encountered in plants-only diets. In the simulated system with no animals, estimated agricultural GHG decreased (28%), but did not fully counterbalance the animal contribution of GHG (49% in this model). This assessment suggests that removing animals from US agriculture would reduce agricultural GHG emissions, but would also create a food supply incapable of supporting the US population's nutritional requirements.

Cobalt (III)-EDTA dissociates and chromium (III)-EDTA is slightly more stable under in vitro reducing conditions comparable to those in the rumen.

Ideal digesta markers used for feeding studies are inert, unabsorbable, and move with the digesta they are intended to mark. Both chromium (III) and cobalt (III) salts of EDTA (CrEDTA and CoEDTA, respectively) are used as markers of liquid digesta in dairy cattle research. A small portion is absorbed and excreted in urine, but the markers are assumed to remain unreactive and as inert salts in the digesta and animal. The degree to which these colored salts remain bound in solution can be estimated through spectrophotometric measurement at their wavelength (λ) of peak absorbance. The objective of this in vitro study was to evaluate whether CrEDTA and CoEDTA dissociate under reducing conditions that could be experienced in the rumen. In a completely randomized design with 2 replicate analytical runs and samples in duplicate within run, approximately 26 mg/L Cr from CrEDTA or Co from CoEDTA was incubated in a 26-mL reaction volume containing 20 mL of Goering and Van Soest medium without tryptone, 3 mL of CoEDTA or CrEDTA solutions, or water (reagent blanks), and 3.0 mL of a combination of distilled water with 0, 0.25, 0.50, 0.75, or 1.00 mL of reducing solution (RedSol). After incubation for 0.5 h at 39°C, absorbance was read at λ = 535, 465, and 560 nm, the peak λ for EDTA salts of Co(III), Co(II), and Cr(III), respectively. Mean reagent blank values were subtracted from CoEDTA and CrEDTA data. The absorbance data at peak λ were analyzed by marker in models that included RedSol with analytical run as a random variable. Contrasts were used to detect linear through quartic effects of RedSol. Samples with RedSol had redox potentials of -250 to -328 mV, which are within the range of reported ruminal measures. As RedSol increased, CoEDTA showed a linear decline of 75% in ABS at 535 nm and a quadratic 4-fold increase followed by a 60% decline at 465 nm. These responses indicate a reduction of Co(III) to Co(II) and subsequent dissociation of Co(II)EDTA. The absorbance of CrEDTA at 560 nm showed a tendency for an 8% linear decrease as RedSol increased. Wavescans from λ = 330 to 700 nm showed CrEDTA retaining its characteristic 2-peak pattern as RedSol increased, whereas CoEDTA curves deformed entirely. We conclude that CoEDTA is not a stable, inert digesta marker under reducing conditions achievable in the rumen and is therefore unsuitable for use in studies with ruminants. Reexamination of the suitability of available liquid digesta markers is advised.

Stability of the liquid markers chromium (III) and cobalt (III)-EDTA in autoclaved, clarified rumen fluid.

An ideal digesta marker for use in feeding studies is inert, unabsorbable, and moves with the portion of the digesta it is intended to mark. Both chromium (III) and cobalt (III) salts of EDTA (CrEDTA and CoEDTA, respectively) have been used as digesta liquid markers in studies with dairy cattle. Although a small portion of these markers is known to be absorbed and excreted in urine, the markers are assumed to remain ionically bound in the digesta. The degree to which these salts remain bound in solution can be determined through spectrophotometric measurement at the wavelength (λ) of peak absorbance of these colored compounds. The objective of this study was to evaluate whether CrEDTA and CoEDTA dissociate during incubation in autoclaved, clarified rumen fluid (ACRF), as indicated by changes in absorbance. In a complete block design with separate replicated analytical runs, approximately 40 mg/L of Cr from CrEDTA or Co from CoEDTA were incubated in 2 separate preparations of ACRF from 2 lactating Holstein cows, in water (CrEDTA), or in MES buffer (CoEDTA), and appropriate reagent blanks. Solution pH were approximately 6.0. Samples were incubated for 24 h at 39°C with absorbance measurements recorded at 0, 1, 2, 4, 6, 22, and 24. The CrEDTA was measured at λ = 541 nm, CoEDTA at λ = 535 nm, and both were measured with wavescans of λ = 330 to 700 nm. At their peak λ, both CrEDTA in water and CoEDTA in MES buffer maintained absorbance values throughout the incubation, whereas, in ACRF, CrEDTA absorbance decreased by 9% at 0 h, and by up to 14% by 24 h; CoEDTA showed a gradual decline over time, with approximately 4% loss in absorbance by 24 h. Responses differed by ACRF preparation. Both markers in ACRF showed increases and decreases over time in absorbance at λ = 330 and 380 nm, though the changes were more marked for CrEDTA; markers not in ACRF showed no change in absorbance at these λ. Changes in the absorbance values at λ = 330 and 380 nm suggest that soluble phenolic compounds may be involved in the exchange of metals with EDTA, but that does not preclude involvement of other components in rumen fluid. Both CrEDTA and CoEDTA incubated in ACRF showed declines over time in absorbance at their peak λ, suggesting that the metals dissociated from EDTA. The apparent dissociation indicates that these liquid markers are not inert as had been assumed.

A 100-Year Review: Carbohydrates-Characterization, digestion, and utilization.

Our knowledge of the role of carbohydrates in dairy cattle nutrition has advanced substantially in the 100 years of the publication of the Journal of Dairy Science. In this review, we trace the history of scientific investigation and discovery from crude fiber, nitrogen-free extract, and "unidentified factors" to our present analytical schemes and understanding of ruminal and whole-animal utilization and effects of dietary carbohydrates. Historically, advances in research and new feeding standards occurred in parallel with and fostered by new methods of analysis. The 100 years of research reviewed here has bequeathed to us an impressive legacy of information, which we will continue to grow.

Physically adjusted neutral detergent fiber system for lactating dairy cow rations. I: Deriving equations that identify factors that influence effectiveness of fiber.

Physically effective neutral detergent fiber (peNDF) is the fraction of neutral detergent fiber (NDF) that stimulates chewing activity and contributes to the floating mat of large particles in the rumen. Multiplying dietary NDF by particle size has been used as an estimate of peNDF. In re-evaluating the concept of peNDF, we compared the use of peNDF as dietary NDF × particle size with the use of individual NDF and particle size descriptors (physically adjusted NDF; paNDF) when used with other physical and chemical diet descriptors to predict dry matter (DM) intake (DMI), rumination time, and ruminal pH in lactating dairy cows. The purpose is to ultimately use these equations to estimate diet adequacy to maintain ruminal conditions. Each response variable had 8 models in a 2 (peNDF, paNDF) × 2 (diet, diet and ruminal factors) × 2 (DM, as fed basis) factorial arrangement. Particle size descriptors were those determined with the Penn State Particle Separator. Treatment means (n = 241) from 60 publications were used in backward elimination multiple regression to derive models of response variables. When available, peNDF terms entered equations. Models containing peNDF terms had similar or lower unadjusted concordance correlation coefficients (an indicator of similar or lower accuracy and precision) than did models without peNDF terms. The peNDF models for rumen pH did not differ substantially from paNDF models. This suggests that peNDF can account for some variation in ruminal pH; however, overt advantages of peNDF were not apparent. Significant variables that entered the models included estimated mean particle size; as fed or DM proportions retained on 19- and 8-mm sieves of the Penn State Particle Separator; DMI; dietary concentrations of forage; forage NDF; CP; starch; NDF; rumen-degraded starch and rumen-degraded NDF; and the interaction terms of starch × mean particle size, acid detergent fiber/NDF, and rumination time/DMI. Many dietary factors beyond particle size and NDF were identified as influencing the response variables. In conclusion, these results appear to justify the development of a modeling approach to integrate individual physical and chemical factors to predict effects on factors affecting rumen conditions.

Physically adjusted neutral detergent fiber system for lactating dairy cow rations. II: Development of feeding recommendations.

The objective of this work was to leverage equations derived in a meta-analysis into an ensemble modeling system for estimating dietary physical and chemical characteristics required to maintain desired rumen conditions in lactating dairy cattle. Given the availability of data, responsiveness of ruminal pH to animal behaviors, and the chemical composition and physical form of the diet, mean ruminal pH was chosen as the primary rumen environment indicator. Physically effective fiber (peNDF) is defined as the fraction of neutral detergent fiber (NDF) that stimulates chewing activity and contributes to the floating mat of large particles in the rumen. The peNDF of feedstuffs is typically estimated by multiplying the NDF content by a particle size measure, resulting in an estimated index of effectiveness. We hypothesized that the utility of peNDF could be expanded and improved by dissociating NDF and particle size and considering other dietary factors, all integrated into a physically adjusted fiber system that can be used to estimate minimum particle sizes of TMR and diet compositions needed to maintain ruminal pH targets. Particle size measures of TMR were limited to those found with the Penn State particle separator (PSPS). Starting with specific diet characteristics, the system employed an ensemble of models that were integrated using a variable mixture of experts approach to generate more robust recommendations for the percentage of dietary DM material that should be retained on the 8-mm sieve of a PSPS. Additional continuous variables also integrated in the physically adjusted fiber system include the proportion of material (dry matter basis) retained on the 19- and 8-mm sieves of the PSPS, estimated mean particle size, the dietary concentrations of forage, forage NDF, starch, and NDF, and ruminally degraded starch and NDF. The system was able to predict that the minimum proportion of material (dry matter basis) retained on the 8-mm sieve should increase with decreasing forage NDF or dietary NDF. Additionally, the minimum proportion of dry matter material on the 8-mm sieve should increase with increasing dietary starch. Results of this study agreed with described interrelationships between the chemical and physical form of diets fed to dairy cows and quantified the links between NDF intake, diet particle size, and ruminal pH. Feeding recommendations can be interpolated from tables and figures included in this work.

Transient changes in milk production efficiency and bacterial community composition resulting from near-total exchange of ruminal contents between high- and low-efficiency Holstein cows.

The objectives of this study were to determine if milk production efficiency (MPE) is altered by near-total exchange of ruminal contents between high- (HE) and low-MPE (LE) cows and to characterize ruminal bacterial community composition (BCC) before exchange and over time postexchange. Three pairs of ruminally cannulated, third-lactation cows were selected whose MPE (energy-corrected milk per unit of dry matter intake) differed over their first 2 lactations. Approximately 95% of ruminal contents were exchanged between cows within each pair. Ruminal pH and volatile fatty acid (VFA) profiles, along with BCC (characterized by sequencing of the variable 4 region of 16S rRNA genes), were assessed just before feeding on d -8, -7, -5, -4, -1, 1, 2, 3, 7, 10, 14, 21, 28, 35, 42, and 56, relative to the exchange date. High-MPE cows had higher total ruminal VFA concentrations, higher molar percentages of propionate and valerate, and lower molar percentages of acetate and butyrate than did LE cows, and re-established these differences 1 d after contents exchange. Across all LE cows, MPE increased during 7 d postexchange but declined thereafter. Two of the 3 HE cows displayed decreases in MPE following introduction of the ruminal contents from the corresponding LE cow, but MPE increased in the third HE cow, which was determined to be an outlier. For all 6 cows, both liquid- and solids-associated BCC differed between individuals within a pair before contents exchange. Upon exchange, BCC of both phases in all 3 pairs was more similar to that of the donor inoculum than to preexchange host BCC. For 5 of 6 cows, the solids-associated community returned within 10 d to more resemble the preexchange community of that host than that of the donor community. Individual variability before the exchange was greater in liquids than in solids, as was the variability in response of bacterial communities to the exchange. Individual cows varied in their response, but generally moved toward re-establishment of their preexchange communities by 10 d after contents exchange. By contrast, ruminal pH and VFA profiles returned to preexchange levels within 1 d. Despite the small number of cows studied, the data suggest an apparent role for the ruminal bacterial community as a determinant of MPE.

Technical note: A method for isolating glycogen granules from ruminal protozoa for further characterization.

Evaluation of physical, chemical, and enzymatic hydrolysis characteristics of protozoal glycogen is best performed on a pure substrate to avoid interference from other cell components. A method for isolating protozoal glycogen granules without use of detergents or other potentially contaminating chemicals was developed. Rumen inoculum was incubated anerobically in vitro with glucose. Glycogen-laden protozoa produced in the fermentation, primarily isotrichids, were allowed to sediment in a separatory funnel and were dispensed. The protozoa were processed through repeated centrifugations and sonication to isolate glycogen granules largely free of feed and cellular debris. The final water-insoluble lyophilized product analyzed as 98.3% α-glucan with very rare starch granules and 1.9% protein. Observed losses of glycogen granules during the clean-up process indicate that this procedure should not be used for quantitative assessment of protozoal glycogen from fermentations. Further optimization of this procedure to enhance the amount of glycogen obtained per fermentation may be possible.

Determination of dietary starch in animal feeds and pet food by an enzymatic-colorimetric method: collaborative study.

Starch, glycogen, maltooligosaccharides, and other α-1,4- and α-1,6-linked glucose carbohydrates, exclusive of resistant starch, are collectively termed "dietary starch". This nutritionally important fraction is increasingly measured for use in diet formulation for animals as it can have positive or negative effects on animal performance and health by affecting energy supply, glycemic index, and formation of fermentation products by gut microbes. AOAC Method 920.40 that was used for measuring dietary starch in animal feeds was invalidated due to discontinued production of a required enzyme. As a replacement, an enzymatic-colorimetric starch assay developed in 1997 that had advantages in ease of sample handling and accuracy compared to other methods was considered. The assay was further modified to improve utilization of laboratory resources and reduce time required for the assay. The assay is quasi-empirical: glucose is the analyte detected, but its release is determined by run conditions and specification of enzymes. The modified assay was tested in an AOAC collaborative study to evaluate its accuracy and reliability for determination of dietary starch in animal feedstuffs and pet foods. In the assay, samples are incubated in screw cap tubes with thermostable α-amylase in pH 5.0 sodium acetate buffer for 1 h at 100°C with periodic mixing to gelatinize and partially hydrolyze α-glucan. Amyloglucosidase is added, and the reaction mixture is incubated at 50°C for 2 h and mixed once. After subsequent addition of water, mixing, clarification, and dilution as needed, free + enzymatically released glucose are measured. Values from a separate determination of free glucose are subtracted to give values for enzymatically released glucose. Dietary starch equals enzymatically released glucose multiplied by 162/180 (or 0.9) divided by the weight of the as received sample. Fifteen laboratories that represented feed company, regulatory, research, and commercial feed testing laboratories analyzed 10 homogenous test materials representing animal feedstuffs and pet foods in duplicate using the dietary starch assay. The test samples ranged from 1 to 70% in dietary starch content and included moist canned dog food, alfalfa pellets, distillers grains, ground corn grain, poultry feed, low starch horse feed, dry dog kibbles, complete dairy cattle feed, soybean meal, and corn silage. The average within-laboratory repeatability SD (sr) for percentage dietary starch in the test samples was 0.49 with a range of 0.03 to 1.56, and among-laboratory repeatability SDs (sR) averaged 0.96 with a range of 0.09 to 2.69. The HorRat averaged 2.0 for all test samples and 1.9 for test samples containing greater than 2% dietary starch. The HorRat results are comparable to those found for AOAC Method 996.11, which measures starch in cereal products. It is recommended that the dietary starch method be accepted for Official First Action status.

Dietary starch source and protein degradability in diets containing sucrose: effects on ruminal measures and proposed mechanism for degradable protein effects.

A feeding study was conducted to evaluate ruminal effects of starch source (STA) and rumen-degradable dietary protein (RDP) in diets with added sucrose. The experimental design was an incomplete Latin square with three 21-d periods, 8 ruminally cannulated lactating cows, and a 2 × 2 factorial arrangement of treatments. Treatments were STA (dry ground corn or high-moisture corn) as more slowly and more rapidly fermenting starch sources, respectively, and relative amount of RDP (+RDP: added protein from soybean meal; -RDP: heat-treated expeller soybean product partially substituted for soybean meal). Diets were formulated to be isonitrogenous and similar in starch and neutral detergent fiber concentrations. Dry matter (DM) intake was 1 kg greater with +RDP compared with -RDP diets. For ruminal digesta measures made 2 h postfeeding, weight of digesta DM was unaffected by treatment; total kilograms of wet digesta and kilograms of liquid tended to be greater with +RDP than with -RDP, and no effect was observed of STA × RDP. Digesta DM percentage was greater with -RDP than with +RDP. At 2 h postfeeding, ruminal pool sizes (mol) of lactate and total AA were larger and those of total organic acids (OA) and ammonia tended to be larger with +RDP than with -RDP; no effects of STA or STA × RDP were detected. Rumen-degradable protein effects on lactate and OA pool sizes may be due to a protein-mediated increase in fermentation rate of carbohydrate. Organic acid concentrations at 2 h postfeeding did not show the same response pattern or significance as the pool size data; high-moisture corn tended to be greater than dry ground corn and no effect was observed for RDP or STA × RDP. Concentration and pool size for OA were more weakly correlated [coefficient of determination (R(2)) = 0.66] than was the case for other ruminal analytes (R(2) >0.80). Organic acid pool size and kilograms of digesta liquid were strongly correlated (R(2) = 0.79), whereas concentration and kilograms of liquid were much less so (R(2) = 0.21). The correlation of OA moles with kilograms of liquid likely relates to the homeostatic mechanism of water flux across the rumen wall to reduce the osmotic gradient with blood as intraruminal moles of solute change. This action compresses the range of ruminal OA concentrations. With kilograms of ruminal liquid differing across individual measurements, the ruminal OA concentration data are not on the equivalent basis required to be reliably useful for assessing the effect of treatments. Further evaluation of protein effects on carbohydrate fermentation and of methods that allow accurate comparison of treatments for their effect on ruminal OA production are warranted.

Maternal immune activation as an epidemiological risk factor for neurodevelopmental disorders: Considerations of timing, severity, individual differences, and sex in human and rodent studies.

Epidemiological evidence suggests that one's risk of being diagnosed with a neurodevelopmental disorder (NDD)-such as autism, ADHD, or schizophrenia-increases significantly if their mother had a viral or bacterial infection during the first or second trimester of pregnancy. Despite this well-known data, little is known about how developing neural systems are perturbed by events such as early-life immune activation. One theory is that the maternal immune response disrupts neural processes important for typical fetal and postnatal development, which can subsequently result in specific and overlapping behavioral phenotypes in offspring, characteristic of NDDs. As such, rodent models of maternal immune activation (MIA) have been useful in elucidating neural mechanisms that may become dysregulated by MIA. This review will start with an up-to-date and in-depth, critical summary of epidemiological data in humans, examining the association between different types of MIA and NDD outcomes in offspring. Thereafter, we will summarize common rodent models of MIA and discuss their relevance to the human epidemiological data. Finally, we will highlight other factors that may interact with or impact MIA and its associated risk for NDDs, and emphasize the importance for researchers to consider these when designing future human and rodent studies. These points to consider include: the sex of the offspring, the developmental timing of the immune challenge, and other factors that may contribute to individual variability in neural and behavioral responses to MIA, such as genetics, parental age, the gut microbiome, prenatal stress, and placental buffering.

A method for the selective depletion of microglia in the dorsal hippocampus in the juvenile rat brain.

BACKGROUND: To understand the role of microglia in brain function and development, methods have emerged to deplete microglia throughout the brain. Liposome-encapsulated clodronate (LEC) can be infused into the brain to deplete microglia in a brain-region and time-specific manner. NEW METHOD: This study validates methodology to deplete microglia in the rat dorsal hippocampus (dHP) during a specific period of juvenile development. Stereotaxic surgery was performed to infuse LEC at postnatal day (P) 16 or 19 into dHP. Rat brains were harvested at various ages to determine specificity of infusion and duration of depletion. RESULTS: P19 infusion of LEC into dHP with a 27G syringe depleted microglia in dHP subregions CA1, dentate gyrus (DG), and CA3 from P24-P30. There was also evidence of depletion in parietal cortex above the infusion site. P16 infusion of LEC with a 32 G syringe depleted microglia only in dHP subregions CA1 and DG from P21-P40. COMPARISON WITH EXISTING METHOD(S): Previous methods have infused LEC intra-hippocampally in adult rats or intra-cerebroventricularly in neonatal rats. This study is the first to publish methodology to deplete microglia in a brain-region specific manner during juvenile rat development. CONCLUSIONS: The timing of LEC infusion during the juvenile period can be adjusted to achieve maximal microglia depletion by a specific postnatal day. A 27G needle results in LEC backflow during the infusion, but also allows LEC to reach all subregions of dHP. Infusion with a 32 G needle prevents backflow during infusion, but results in a more local spread of LEC within dHP.

Comparison of methods for glycogen analysis of in vitro fermentation pellets produced with strained ruminal inoculum.

Microbial glycogen measurement is used to account for fates of carbohydrate substrates. It is commonly applied to washed cells or pure cultures which can be accurately subsampled, allowing the use of smaller sample sizes. However, the nonhomogeneous fermentation pellets produced with strained rumen inoculum cannot be accurately subsampled, requiring analysis of the entire pellet. In this study, two microbial glycogen methods were compared for analysis of such fermentation pellets: boiling samples for 3h in 30% KOH (KOH) or for 15min in 0.2M NaOH (NaOH), followed by enzymatic hydrolysis with α-amylase and amyloglucosidase, and detection of released glucose. Total α-glucan was calculated as glucose×0.9. KOH and NaOH did not differ in the α-glucan detected in fermentation pellets (29.9 and 29.6mg, respectively; P=0.61). Recovery of different control α-glucans was also tested using KOH, NaOH, and a method employing 45min of bead beating (BB). For purified beef liver glycogen (water-soluble) recovery, BB (95.0%)>KOH (91.4%)>NaOH (87.4%; P<0.05), and for wheat starch (water-insoluble granules) recovery, NaOH (96.9%)>BB (93.8%)>KOH (91.0%; P<0.05). Recovery of isolated protozoal glycogen (water-insoluble granules) did not differ among KOH (87.0%), NaOH (87.6%), and BB (86.0%; P=0.81), but recoveries for all were below 90%. Differences among substrates in the need for gelatinization and susceptibility to destruction by alkali likely affected the results. In conclusion, KOH and NaOH glycogen methods provided comparable determinations of fermentation pellet α-glucan. The tests on purified α-glucans indicated that assessment of recovery in glycogen methods can differ by the control α-glucan selected.

Feed analyses and their interpretation.

Compositional analysis is central to determining the nutritional value of feedstuffs for use in ration formulation. The utility of the values and how they should be used depends on how representative the feed subsample is, the nutritional relevance and analytical variability of the assays, and whether an analysis is suitable to be applied to a particular feedstuff. Commercial analyses presently available for carbohydrates, protein, and fats have improved nutritionally pertinent description of feed fractions. Factors affecting interpretation of feed analyses and the nutritional relevance and application of currently available analyses are discussed.

Feeding, evaluating, and controlling rumen function.

Achieving optimal rumen function requires an understanding of feeds and systems of nutritional evaluation. Key influences on optimal function include achieving good dry matter intake. The function of feeds in the rumen depends on other factors including chemical composition, rate of passage, degradation rate of the feed, availability of other substrates and cofactors, and individual animal variation. This article discusses carbohydrate, protein, and fat metabolism in the rumen, and provides practical means of evaluation of rations in the field. Conditions under which rumen function is suboptimal (ie, acidosis and bloat) are discussed, and methods for control examined.

Gated SPECT perfusion imaging for the simultaneous assessment of myocardial perfusion and ventricular function in the BARI 2D trial: an initial report from the Nuclear Core Laboratory.

BACKGROUND: The BARI 2D (Bypass Angioplasty Revascularization Investigation 2 Diabetes) trial, a National Heart, Lung, and Blood Institute-sponsored study in type 2 diabetic patients with coronary artery disease, completed patient recruitment in March 2005. This trial had a nuclear substudy in addition to many other substudies. METHODS AND RESULTS: After patient enrollment, adenosine gated single photon emission computed tomography perfusion imaging is performed at years 1 and 3. The images are interpreted at the core laboratory. Among the objectives of the nuclear substudy are (1) to determine the impact of the mode of therapy on left ventricular function, extent of ischemia, and scar; (2) to determine the impact of therapy on the progression/regression of ischemia/scar and changes in left ventricular function between years 1 and 3; and (3) to determine the independent and incremental prognostic value of ischemia, scar, and left ventricular function on the primary and secondary endpoints of the trial in the entire patient population and specified subgroups such as women, elderly patients, and minorities. CONCLUSIONS: This article describes the methodology and the initial experience of the nuclear core laboratory in this large multicenter trial and provides a summary of variables that are available for future analysis by the working group.