Low-Protein Diets Differentially Regulate Energy Balance during Thermoneutral and Heat Stress in Cobb Broiler Chicken (Gallus domesticus).

The objective was to assess whether low-protein (LP) diets regulate food intake (FI) and thermogenesis differently during thermoneutral (TN) and heat stress (HS) conditions. Two-hundred-day-old male broiler chicks were weight-matched and assigned to 36 pens with 5-6 chicks/pen. After 2 weeks of acclimation, birds were subjected into four groups (9 pens/group) including (1) a normal-protein diet under TN (ambient temperature), (2) an LP diet under TN, (3) a normal-protein diet under HS (35 °C for 7 h/day), and (4) an LP diet under HS, for 4 weeks. During HS, but not TN, LP tended to decrease FI, which might be associated with a lower mRNA abundance of duodenal ghrelin and higher GIP during HS. The LP group had a higher thermal radiation than NP under TN, but during HS, the LP group had a lower thermal radiation than NP. This was linked with higher a transcript of muscle ß1AR and AMPKα1 during TN, but not HS. Further, LP increased the gene expression of COX IV during TN but reduced COX IV and the sirtuin 1 abundance during HS. The dietary protein content differentially impacted plasma metabolome during TN and HS with divergent changes in amino acids such as tyrosine and tryptophan. Compared to NP, LP had increased abundances of p_Tenericutes, c_Mollicutes, c_Mollicutes_RF9, and f_tachnospiraceae under HS. Overall, LP diets may mitigate the negative outcome of heat stress on the survivability of birds by reducing FI and heat production. The differential effect of an LP diet on energy balance during TN and HS is likely regulated by gut and skeletal muscle and alterations in plasma metabolites and cecal microbiota.

Social and genetic connectivity despite ecological variation in a killer whale network.

Philopatric kin-based societies encourage a narrow breadth of conservative behaviours owing to individuals primarily learning from close kin, promoting behavioural homogeneity. However, weaker social ties beyond kin, and across a behaviourally diverse social landscape, could be sufficient to induce variation and a greater ecological niche breadth. We investigated a network of 457 photo-identified killer whales from Norway (548 encounters in 2008-2021) with diet data available (46 mixed-diet individuals feeding on both fish and mammals, and 411 exclusive fish-eaters) to quantify patterns of association within and between diet groups, and to identify underlying correlates. We genotyped a subset of 106 whales to assess patterns of genetic differentiation. Our results suggested kinship as main driver of social bonds within and among cohesive social units, while diet was most likely a consequence reflective of cultural diffusion, rather than a driver. Flexible associations within and between ecologically diverse social units led to a highly connected network, reducing social and genetic differentiation between diet groups. Our study points to a role of social connectivity, in combination with individual behavioural variation, in influencing population ecology in killer whales.

Diversity and community structure of anaerobic gut fungi in the rumen of wild and domesticated herbivores.

The rumen houses a diverse community that plays a major role in the digestion process in ruminants. Anaerobic gut fungi (AGF) are key contributors to plant digestion in the rumen. Here, we present a global amplicon-based survey of the rumen AGF mycobiome by examining 206 samples from 15 animal species, 15 countries, and 6 continents. The rumen AGF mycobiome was highly diverse, with 81 out of 88 currently recognized AGF genera or candidate genera identified. However, only six genera (Neocallimastix, Orpinomyces, Caecomyces, Cyllamyces, NY9, and Piromyces) were present at >4% relative abundance. AGF diversity was higher in members of the families Antilocapridae and Cervidae compared to Bovidae. Community structure analysis identified a pattern of phylosymbiosis, where host family (10% of total variance) and species (13.5%) partially explained the rumen mycobiome composition. As well, diet composition (9%-19%), domestication (11.14%), and biogeography (14.1%) also partially explained AGF community structure; although sampling limitation, geographic range restrictions, and direct association between different factors hindered accurate elucidation of the relative contribution of each factor. Pairwise comparison of rumen and fecal samples obtained from the same subject (n = 13) demonstrated greater diversity and inter-sample variability in rumen versus fecal samples. The genera Neocallimastix and Orpinomyces were present in higher abundance in rumen samples, while Cyllamyces and Caecomyces were enriched in fecal samples. Comparative analysis of global rumen and feces data sets revealed a similar pattern. Our results provide a global view of AGF community in the rumen and identify patterns of AGF variability between rumen and feces in herbivores Gastrointestinal (GI) tract.IMPORTANCERuminants are highly successful and economically important mammalian suborder. Ruminants are herbivores that digest plant material with the aid of microorganisms residing in their GI tract. In ruminants, the rumen compartment represents the most important location where microbially mediated plant digestion occurs, and is known to house a bewildering array of microbial diversity. An important component of the rumen microbiome is the anaerobic gut fungi (AGF), members of the phylum Neocallimastigomycota. So far, studies examining AGF diversity have mostly employed fecal samples, and little is currently known regarding the identity of AGF residing in the rumen compartment, factors that impact the observed patterns of diversity and community structure of AGF in the rumen, and how AGF communities in the rumen compare to AGF communities in feces. Here, we examined the rumen AGF diversity using an amplicon-based survey targeting a wide range of wild and domesticated ruminants (n = 206, 15 different animal species) obtained from 15 different countries. Our results demonstrate that while highly diverse, no new AGF genera were identified in the rumen mycobiome samples examined. Our analysis also indicate that animal host phylogeny, diet, biogeography, and domestication status could play a role in shaping AGF community structure. Finally, we demonstrate that a greater level of diversity and higher inter-sample variability was observed in rumen compared to fecal samples, with two genera (Neocallimastix and Orpinomyces) present in higher abundance in rumen samples, and two others (Cyllamyces and Caecomyces) enriched in fecal samples. Our results provide a global view of the identity, diversity, and community structure of AGF in ruminants, elucidate factors impacting diversity and community structure of the rumen mycobiome, and identify patterns of AGF community variability between the rumen and feces in the herbivorous GI tract.

Association of glucose metabolism and insulin resistance with feed efficiency and production traits of finishing beef steers.

Increasing nutrient utilization efficiency is an important component of enhancing the sustainability of beef cattle production. The objective of this experiment was to determine the association of glucose metabolism and insulin resistance with dry matter intake (DMI), average daily gain (ADG), gain:feed ratio (G:F), and residual feed intake (RFI). Steers (n = 54; initial body weight = 518 ±â€…27.0 kg) were subjected to an intravenous glucose tolerance test (IVGTT) where glucose was dosed through a jugular catheter and serial blood samples were collected. Three days after the last group's IVGTT, steers began a 63-d DMI and ADG test. Body weight was measured on days 0, 1, 21, 42, 62, and 63, and DMI was measured using an Insentec Roughage Intake Control system (Hokofarm Group, Emmeloord, the Netherlands). To examine relationships between DMI, ADG, G:F, and RFI with IVGTT measurements, Pearson correlations were calculated using Proc Corr of SAS 9.4 (SAS Inst. Inc., Cary, NC). Additionally, cattle were classified based on DMI, ADG, RFI, and G:F, where the medium classification was set as mean ±â€…0.5 SD, the low classification was < 0.5 SD from the mean, and the high classification was > 0.5 SD from the mean. No associations between DMI and IVGTT parameters were observed, and no differences were detected when classifying cattle as having low, medium, or high DMI. Peak insulin concentration in response to the IVGTT tended to be correlated with ADG (r = 0.28; P = 0.07), indicating cattle with greater ADG tend to have a greater insulin release in response to glucose. Glucose nadir concentrations tended to be positively correlated with ADG (r = 0.26; P = 0.10). Additionally, the glucose nadir was greater in high-ADG steers (P = 0.003). The association of greater glucose nadir with high-ADG could indicate that high-ADG steers do not clear glucose as efficiently as low-ADG steers, potentially indicating increased insulin resistance. Further, RFI was not correlated with IVGTT measurements, but low RFI steers had a greater peak glucose concentration (P = 0.040) and tended to have a greater glucose area under the curve (P = 0.09). G:F was correlated with glucose area under the curve (r = 0.33; P = 0.050), glucose nadir (r = 0.35; P = 0.011), and insulin time to peak (r = 0.39; P = 0.010). These results indicate that glucose metabolism and insulin signaling are associated with growth and efficiency, but the molecular mechanisms that drive these effects need to be elucidated.

Feed efficiency is an important component of improving the sustainability of beef production. There is a need to understand how metabolism influences feed efficiency. This experiment aimed to explore the association of glucose metabolism and insulin resistance with feed intake, growth, and efficiency of finishing beef cattle. The results indicate that there is a relationship between insulin resistance and improved efficiency measured as the ratio of growth to feed intake. The findings of this experiment are novel as they show a relationship between insulin resistance and feed efficiency and indicate further research is needed to determine the mechanisms of insulin resistance in ruminants that contribute to improved feed efficiency.

Late Pleistocene stickleback environmental genomes reveal the chronology of freshwater adaptation.

Directly observing the chronology and tempo of adaptation in response to ecological change is rarely possible in natural ecosystems. Sedimentary ancient DNA (sedaDNA) has been shown to be a tractable source of genome-scale data of long-dead organisms1,2,3 and to thereby potentially provide an understanding of the evolutionary histories of past populations.4,5 To date, time series of ecosystem biodiversity have been reconstructed from sedaDNA, typically using DNA metabarcoding or shotgun sequence data generated from less than 1 g of sediment.6,7 Here, we maximize sequence coverage by extracting DNA from ∼50× more sediment per sample than the majority of previous studies1,2,3 to achieve genotype resolution. From a time series of Late Pleistocene sediments spanning from a marine to freshwater ecosystem, we compare adaptive genotypes reconstructed from the environmental genomes of three-spined stickleback at key time points of this transition. We find a staggered temporal dynamic in which freshwater alleles at known loci of large effect in marine-freshwater divergence of three-spined stickleback (e.g., EDA)8 were already established during the brackish phase of the formation of the isolation basin. However, marine alleles were still detected across the majority of marine-freshwater divergence-associated loci, even after the complete isolation of the lake from marine ingression. Our retrospective approach to studying adaptation from environmental genomes of three-spined sticklebacks at the end of the last glacial period complements contemporary experimental approaches9,10,11 and highlights the untapped potential for retrospective "evolve and resequence" natural experiments using sedaDNA.

Divergence and gene flow history at two large chromosomal inversions underlying ecotype differentiation in the long-snouted seahorse.

Chromosomal inversions can play an important role in divergence and reproductive isolation by building and maintaining distinct allelic combinations between evolutionary lineages. Alternatively, they can take the form of balanced polymorphisms that segregate within populations until one arrangement becomes fixed. Many questions remain about how inversion polymorphisms arise, how they are maintained over the long term, and ultimately, whether and how they contribute to speciation. The long-snouted seahorse (Hippocampus guttulatus) is genetically subdivided into geographic lineages and marine-lagoon ecotypes, with shared structural variation underlying lineage and ecotype divergence. Here, we aim to characterize structural variants and to reconstruct their history and suspected role in ecotype formation. We generated a near chromosome-level genome assembly and described genome-wide patterns of diversity and divergence through the analysis of 112 whole-genome sequences from Atlantic, Mediterranean, and Black Sea populations. By also analysing linked-read sequencing data, we found evidence for two chromosomal inversions that were several megabases in length and showed contrasting allele frequency patterns between lineages and ecotypes across the species range. We reveal that these inversions represent ancient intraspecific polymorphisms, one likely being maintained by divergent selection and the other by pseudo-overdominance. A possible selective coupling between the two inversions was further supported by the absence of specific haplotype combinations and a putative functional interaction between the two inversions in reproduction. Lastly, we detected gene flux eroding divergence between inverted alleles at varying levels for the two inversions, with a likely impact on their dynamics and contribution to divergence and speciation.

Ancestry testing of "Old Tom," a killer whale central to mutualistic interactions with human whalers.

Cooperative hunting between humans and killer whales (Orcinus orca) targeting baleen whales was reported in Eden, New South Wales, Australia, for almost a century. By 1928, whaling operations had ceased, and local killer whale sightings became scarce. A killer whale from the group, known as "Old Tom," washed up dead in 1930 and his skeleton was preserved. How these killer whales from Eden relate to other populations globally and whether their genetic descendants persist today remains unknown. We extracted and sequenced DNA from Old Tom using ancient DNA techniques. Genomic sequences were then compared with a global dataset of mitochondrial and nuclear genomes. Old Tom shared a most recent common ancestor with killer whales from Australasia, the North Atlantic, and the North Pacific, having the highest genetic similarity with contemporary New Zealand killer whales. However, much of the variation found in Old Tom's genome was not shared with these widespread populations, suggesting ancestral rather than ongoing gene flow. Our genetic comparisons also failed to find any clear descendants of Tom, raising the possibility of local extinction of this group. We integrated Traditional Custodian knowledge to recapture the events in Eden and recognize that Indigenous Australians initiated the relationship with the killer whales before European colonization and the advent of commercial whaling locally. This study rectifies discrepancies in local records and provides new insight into the origins of the killer whales in Eden and the history of Australasian killer whales.

Technical note: Validation of energy dispersive X-ray fluorescence for determination of indigestible markers in ruminant fecal and rumen fluid samples.

Determination of digestibility and passage rate is important for further understanding of nutrient utilization and thereby aids in improving nutrient utilization efficiency. Titanium dioxide and chromium ethylenediaminetetraacetic acid are commonly used as indigestible markers for determining passage rates of diets to aid in determination of digestibility. Analyzing Ti and Cr involves the use of procedures such as acid digestions, inductively coupled plasma spectroscopy, and atomic absorption. These commonly used methodologies involve hazardous chemicals, destruction of samples, and low sample throughput. The objective of this experiment was to develop and validate an accurate and precise method for measuring both Ti and Cr using energy dispersive X-ray fluorescence (ED-XRF). Energy dispersive X-ray fluorescence is an analytical technique used for analyzing elements in various sample types. The samples were added to the ED-XRF machine and irradiated with X-rays. The intensity of the X-rays emitted (termed fluorescent X-rays) was used for calculation of the concentration of the element. The method for Ti was constructed using fecal samples from cattle consuming three different diet types (finishing diet, dairy lactation diet, and grazing native range pasture). The Cr method was developed for rumen fluid analysis. We compared the machine-calculated concentrations of each element to the concentration calculated by a standard curve. For both the Ti and Cr, the standard curve-calculated value had a lower percent difference overall at 4.56% and 12.59%, respectively, compared to the machine percent difference of 8.35% and 16.38% for Ti and Cr, respectively. To determine accuracy and precision of the method, samples were spiked with various amounts of Ti or Cr and measured for their respective compounds with percent recovery and inter- and intra-assay CV-calculated thereafter. The average recovery for Ti across all diet types was 100.3%, and the recovery for Cr in rumen fluid was 95.7%. The average inter- and intra-assay CV for Ti, across all diet types, were 9.70% and 2.16%, respectively. For Cr, the average inter- and intra-assay CV were 5.42% and 8.45%, respectively. The ED-XRF method requires minimal additional chemicals, is cost-effective, and allows for sample preservation as well as a high throughput of samples. Our results indicate utilization of ED-XRF is an accurate and precise method for determination of Ti in feces and Cr in rumen fluid.

Markers are elements supplied to an animal to help determine the digestibility of various feedstuffs. Studying digestibility allows us to improve how nutrients are used by animals and reduce production costs and environmental impact. The method proposed here uses an X-ray fluorescence machine to analyze the markers with no hazardous chemicals, low labor input, and cost. Even though this equipment utilizes radiation, the construction of the equipment eliminates the exposure of operators to radiation. Training and monitoring will differ between institutions. Traditional methods use hazardous chemicals and have higher cost and labor use. We determined that this is a valid method for analyzing titanium in fecal samples and chromium in rumen fluid.

Ancient dolphin genomes reveal rapid repeated adaptation to coastal waters.

Parallel evolution provides strong evidence of adaptation by natural selection due to local environmental variation. Yet, the chronology, and mode of the process of parallel evolution remains debated. Here, we harness the temporal resolution of paleogenomics to address these long-standing questions, by comparing genomes originating from the mid-Holocene (8610-5626 years before present, BP) to contemporary pairs of coastal-pelagic ecotypes of bottlenose dolphin. We find that the affinity of ancient samples to coastal populations increases as the age of the samples decreases. We assess the youngest genome (5626 years BP) at sites previously inferred to be under parallel selection to coastal habitats and find it contained coastal-associated genotypes. Thus, coastal-associated variants rose to detectable frequencies close to the emergence of coastal habitat. Admixture graph analyses reveal a reticulate evolutionary history between pelagic and coastal populations, sharing standing genetic variation that facilitated rapid adaptation to newly emerged coastal habitats.

Temporal progression of anaerobic fungal communities in dairy calves from birth to maturity.

Establishment of microbial communities in neonatal calves is vital for their growth and overall health. While this process has received considerable attention for bacteria, our knowledge on temporal progression of anaerobic gut fungi (AGF) in calves is lacking. Here, we examined AGF communities in faecal samples from six dairy cattle collected at 24 different time points during the pre-weaning (days 1-48), weaning (days 48-60), and post-weaning (days 60-360) phases. Quantitative polymerase chain reaction indicated that AGF colonisation occurs within 24 h after birth, with loads slowly increasing during pre-weaning and weaning, then drastically increasing post-weaning. Culture-independent amplicon surveys identified higher alpha diversity during pre-weaning/weaning, compared to post-weaning. AGF community structure underwent a drastic shift post-weaning, from a community enriched in genera commonly encountered in hindgut fermenters to one enriched in genera commonly encountered in adult ruminants. Comparison of AGF community between calves day 1 post-birth and their mothers suggest a major role for maternal transmission, with additional input from cohabitating subjects. This distinct pattern of AGF progression could best be understood in-light of their narrower niche preferences, metabolic specialisation, and physiological optima compared to bacteria, hence eliciting a unique response to changes in feeding pattern and associated structural GIT development during maturation.

Patterns and determinants of the global herbivorous mycobiome.

Despite their role in host nutrition, the anaerobic gut fungal (AGF) component of the herbivorous gut microbiome remains poorly characterized. Here, to examine global patterns and determinants of AGF diversity, we generate and analyze an amplicon dataset from 661 fecal samples from 34 mammalian species, 9 families, and 6 continents. We identify 56 novel genera, greatly expanding AGF diversity beyond current estimates (31 genera and candidate genera). Community structure analysis indicates that host phylogenetic affiliation, not domestication status and biogeography, shapes the community rather than. Fungal-host associations are stronger and more specific in hindgut fermenters than in foregut fermenters. Transcriptomics-enabled phylogenomic and molecular clock analyses of 52 strains from 14 genera indicate that most genera with preferences for hindgut hosts evolved earlier (44-58 Mya) than those with preferences for foregut hosts (22-32 Mya). Our results greatly expand the documented scope of AGF diversity and provide an ecologically and evolutionary-grounded model to explain the observed patterns of AGF diversity in extant animal hosts.

Effects of adding ruminal propionate on dry matter intake and glucose metabolism in steers fed a finishing ration.

The objective of this experiment was to determine if supplying additional propionate to the rumen alters dry matter intake (DMI), feeding behavior, glucose metabolism, and rumen fluid metabolites in steers fed a finishing diet. Ruminally cannulated steers (n = 6) were fed a finishing diet ad libitum. Steers were randomly assigned to one of three treatments in a 3 × 6 Latin rectangle design with three 15 d periods. Treatments of no Ca propionate (Control), 100 g/d (Low), or 300 g/d (High) were ruminally dosed twice daily. Individual intake was measured using an Insentec feeding system. Pre-feeding blood samples were collected on day 7 and rumen fluid samples were collected on day 13. An intravenous glucose tolerance test (IVGTT) was conducted on day 14 and liver biopsies were collected on day 15. Liver samples were analyzed for expression of genes involved in gluconeogenesis. Data were analyzed using a mixed model with period, treatment, day, and their interaction included, with day and minute within period as a repeated measure and steer as a random effect. Meal size (P = 0.049), meal frequency (P = 0.046), and DMI (P < 0.001) were decreased in High steers. Day 7 plasma glucose (P = 0.23) and lactate (P = 0.47) were not affected by treatment, but insulin was decreased (P = 0.008) and non-esterified fatty acids were increased (P = 0.044) in the High treatment compared with the Control. Rumen fluid lactate was decreased (P = 0.015) in the High treatment compared with the Low treatment. Total VFA concentrations did not differ (P = 0.88) between treatments. There was treatment × time interaction for proportions of acetate and propionate (P < 0.001) and the acetate:propionate ratio (P = 0.005). The effect on acetate was due to a decrease in the High treatment 2 h after dosing the treatment. Propionate proportions were greater in the High treatment than the Control at all time points and differed from the Low except at 0 h. Propionate treatments had no major effects on the glucose and insulin parameters observed in the IVGTT other than a tendency (P = 0.09) for an increased insulin time to peak. These data indicate that exogenous propionate decreases DMI but the decrease in propionate from fermentation due to reduced DMI might negate the supply of exogenous propionate in VFA supply to the animal. Mechanisms other than hepatic oxidation of propionate might be responsible for DMI regulation.

Propionate metabolism by the liver is thought to be a key regulator of appetite and feed intake of animals, including cattle. Previous research has shown that providing propionate to the rumen of cattle decreases feed intake. Propionate is also a major contributor to glucose for cattle to use as an energy source for growth and maintenance. In this experiment, it was hypothesized that increasing ruminal propionate would depress feed intake and decrease insulin sensitivity. Supplying 300 g of propionate a day to the rumen decreased feed intake and increased the proportion of propionate in the rumen fluid of steers. However, when propionate production was calculated based on feed intake, there was likely no difference in propionate supply to the animal. The lack of increase in propionate supply to the animal could explain the lack of effect on glucose metabolism, insulin sensitivity, and liver gene expression. The lack of an increase in propionate also indicates that the effect of propionate on feed intake could be due to alternative mechanisms than liver metabolism of propionate.

"Type D" killer whale genomes reveal long-term small population size and low genetic diversity.

Genome sequences can reveal the extent of inbreeding in small populations. Here, we present the first genomic characterization of type D killer whales, a distinctive eco/morphotype with a circumpolar, subantarctic distribution. Effective population size is the lowest estimated from any killer whale genome and indicates a severe population bottleneck. Consequently, type D genomes show among the highest level of inbreeding reported for any mammalian species (FROH ≥ 0.65). Detected recombination cross-over events of different haplotypes are up to an order of magnitude rarer than in other killer whale genomes studied to date. Comparison of genomic data from a museum specimen of a type D killer whale that stranded in New Zealand in 1955, with 3 modern genomes from the Cape Horn area, reveals high covariance and identity-by-state of alleles, suggesting these genomic characteristics and demographic history are shared among geographically dispersed social groups within this morphotype. Limitations to the insights gained in this study stem from the nonindependence of the 3 closely related modern genomes, the recent coalescence time of most variation within the genomes, and the nonequilibrium population history which violates the assumptions of many model-based methods. Long-range linkage disequilibrium and extensive runs of homozygosity found in type D genomes provide the potential basis for both the distinctive morphology, and the coupling of genetic barriers to gene flow with other killer whale populations.

Meta-Regression to Develop Predictive Equations for Urinary Nitrogen Excretion of Lactating Dairy Cows.

Dairy cows' urinary nitrogen (N) excretion (UN; g/d) represents a significant environmental concern due to their contribution to nitrate leaching, nitrous oxide (a potent greenhouse gas), and ammonia emissions (contributor to N deposition). The first objective of the current study was to determine the adequacy of existing models to predict UN from total mixed ration (TMR)-fed and fresh forage (FF)-fed cows. Next, we aimed to develop equations to predict UN based on animal factors [milk urea nitrogen (MUN; mg/dL) and body weight (BW, kg)] and to explore how these equations are improved when dietary factors, such as diet type, dry matter intake (DMI), or dietary characteristics [neutral detergent fiber (NDF) and crude protein (CP) content], are considered. A dataset was obtained from 51 published experiments composed of 174 treatment means. The whole dataset was used to evaluate the mean and linear biases of three existing equations including diet type as an interaction term; all models had significant linear and mean biases and two of the three models had poor predictive capabilities as indicated by their large relative prediction error (RPE; root mean square error of prediction as a percent of the observed mean). Next, the complete data set was split into training and test sets, which were used to develop and to evaluate new models, respectively. The first model included MUN and BW, and there was a significant interaction between diet type and the coefficients. This model had the worst 1:1 agreement [Lin's concordance correlation coefficient (CCC) = 0.50] and largest RPE (24.7%). Models that included both animal and dietary factors performed the best, and when included in the model, the effect of diet type was no longer significant (p > 0.10). These models all had very good agreement (CCC ≥ 0.86) and relatively low RPE (≤13.1%). This meta-analysis developed precise and accurate equations to predict UN from dairy cows in both confined and pasture-based systems.

Association of maternal temperament and offspring disposition on growth performance.

Animal behavior is complex and varies in definition, depending upon specific traits under observation. Temperament is one component of behavior, that in cattle, is described as the level of fearfulness to a novel or threatening environment. Temperament is a heritable trait which is important since aggressiveness and docility contribute to reproductive success, growth, and carcass quality. We observed maternal temperament at calving and the subsequent influence, if any, on offspring disposition at weaning and their effects collectively on growth performance and carcass traits. Maternal behaviors at calving were observed at four locations within the University of Arkansas system. Cows were assigned a maternal disposition score (MDS) at calving; a scale from 1 to 5 in which aggression decreases. At weaning, calves were assigned a chute score (CS); a scale from 1 to 6 in which aggression increases. Both scoring systems have been previously established. Blood was collected during the 56-d backgrounding period postweaning for blood glucose analysis. Data were analyzed using GLIMMIX procedures of SAS (α = 0.05). The relationship between the two scoring systems was determined with a Pearson correlation (P = 0.22). Animal was the experimental unit and blocked by location for all dependent variables. Location, sex, diet, and MDS were included in the class as covariables for all growth performance and carcass data related to CS. Cows that were more aggressive birthed heavier calves (P < 0.01) compared to indifferent cows. Calves born to cows with either very aggressive or very attentive (MDS of 2 or 3, respectively) scores were heavier upon feedlot entry (P = 0.03) compared to those from indifferent or apathetic cows (MDS of 4 or 5, respectively). Calves defined as nervous and restless (CS of 3 and 2, respectively) were heavier at weaning compared to docile calves (P < 0.01). Restless calves were heavier compared to nervous calves upon arrival and exiting the feedlot (P ≤ 0.01). Calves that were docile at weaning had greater marbling compared to calves that were restless (P ≤ 0.01). Calves that were restless at weaning had greater lean muscle area compared to calves that were nervous (P = 0.05). No definitive relationship was determined between dam and calf temperament. However, the results suggest temperament does impact growth performance and carcass traits but whether the influence comes from the dam or calf temperament, specifically, remains unanswered.

Dam parity affects fetal growth, placental size, and neonatal metabolism in spring-born beef calves.

To determine effects of dam parity on perinatal nutrient availability in beef cattle, data and samples were collected from 18 primiparous and 35 multiparous spring-calving Sim-Angus dams and their calves. Time to stand was recorded and neonatal vigor assessed. Jugular blood was collected from a subset of calves at 0 (post-standing and pre-suckling) 6, 12, 24, 48, and 72 h of age, and blood chemistry panels were completed. Expelled placentas were dissected, dried, and weighed. Prepartum maternal circulating glucose, non-esterified fatty acids (NEFA), triglycerides, and urea N were analyzed. All statistical models included the fixed effect of dam parity, and calf sex (when P ≤ 0.25) was included for calf and placental variables. Effects of sampling hour, and parity × hour were included for calf metabolites over time using repeated measures. Multiparous dams had greater body weight prepartum (P < 0.001) but similar (P = 0.25) body condition score. Maternal circulating urea N and triglycerides were greater (P ≤ 0.05) in multiparous dams pre-calving. Calves born to primiparous dams weighed 10% less (P ≤ 0.04) at birth with smaller (P ≤ 0.01) heart and abdominal girths. Cotyledonary, intercotyledonary, and total placental masses were less (P ≤ 0.05) for primiparous dams. Dam parity did not affect (P ≥ 0.58) calf time to stand, vigor score at 10 min, or rectal temperature. Serum glucose was greater (P = 0.03) at 0 h but less (P ≤ 0.04) at all other hours in calves from primiparous dams. Calves from primiparous dams had greater (P ≤ 0.02) serum NEFA at 6, 12, and 24 h although plasma triglycerides were greater (P < 0.001) at 6 h. Calves from primiparous dams had greater (P ≤ 0.04) serum urea N at 12 h and creatinine at 12 and 24 h. Plasma insulin was greater (P ≤ 0.04) in calves from multiparous dams at 12, 48, and 72 h, but parity did not affect (P ≥ 0.18) serum total protein or plasma cortisol. Serum aspartate aminotransferase was greater (P ≤ 0.04) at 6 and 24 h, creatine kinase was greater at 24 h, and gamma-glutamyl transpeptidase was less (P ≤ 0.04) at 6, 12, and 24 h, for calves from primiparous dams. Calves born to primiparous dams had greater (P ≤ 0.02) total bilirubin and direct bilirubin at 12 and 24 h. Data indicate that calves born to first-parity heifers had decreased perinatal nutrient availability, resulting in reduced fetal and placental growth, as well as greater energy reserve mobilization and metabolic indicators of stress as neonates.

Approximately two-thirds of beef calf deaths prior to weaning occur within the first 3 wk after birth. The goal to have heifers produce their first calf by 2 yr of age likely contributes to factors that limit nutrients available for fetuses and calves immediately after birth. However, little is known about differences in heifers (first parity) and cows (later parities) regarding factors affecting calf resilience, such as fetal growth and calf metabolism shortly after birth. Our data show that calves born to first-parity heifers had altered nutrient availability, demonstrated through smaller placentas, lower birth weights, and altered metabolites in early life. Although calves had similar vigor and ability to maintain body temperature, calves born to first-parity heifers had to mobilize more energy and had lower insulin during the first 3 d post-birth. Calves born to first-parity heifers had greater indicators of stress during the first 72 h of life not associated with calving difficulties. Overall, these effects may have increased morbidity and mortality of calves born to first-parity heifers if they were in a less intensively-managed system. Better understanding of challenges faced by calves born to first-parity dams provides opportunities for their improved management.

Retained energy in lactating beef cows; effects on maintenance energy requirement and voluntary feed intake.

The objectives of these experiments were to determine the relationship between maintenance requirements and energy partitioned to maternal tissue or milk production in limit-fed Angus cows and to determine the relationship between retained energy during the lactation period to dry-period voluntary forage intake (VDMI). Twenty-four mature fall-calving Angus cows were used in a 79-d study during late lactation to establish daily metabolizable energy required for maintenance (MEm). Cows were individually fed daily a mixed diet (2.62 Mcal MEl/kg, 18.2% crude protein) to meet energy and protein requirements of 505 kg beef cows producing 8.2 kg milk daily. If cow BW changed by ±9 kg from initial BW, daily feed intake was adjusted to slow BW loss or reduce BW gain. Milk yield and composition were determined on 3 occasions throughout the study. Maintenance was computed as metabolizable energy intake minus retained energy assigned to average daily maternal tissue energy change, average daily milk energy yield, and average daily energy required for pregnancy. After calves were weaned, cows were fed a low-quality grass hay diet (8.2% crude protein, 65% NDF) and VDMI was measured for 21 days. Lactation maintenance energy was 83% the default value recommended by NASEM (2016. Nutrient Requirements of Beef Cattle: Eighth Revised Edition.) for lactating Angus cows. Increasing lactation-period retained energy (decreasing BW loss and increasing milk energy yield) was associated with lower maintenance energy requirements (P < 0.01; R 2 = 0.92). Increased residual daily gain during lactation was associated with lower lactation maintenance energy requirements (P = 0.05; R 2 = 0.17). Post-weaning VDMI was not related to late-lactation milk energy production, although sensitive to lactation period BCS and BW loss. These results contradict previous reports, suggesting that maintenance requirements increase with increasing milk yield.

TECHNICAL NOTE: Analysis of volatile fatty acids in rumen fluid by gas chromatography mass spectrometry using a dimethyl carbonate extraction.

Analysis of rumen fluid volatile fatty acids (VFA) is typically conducted by injecting acidified aqueous rumen fluid into a gas chromatograph (GC) with a flame ionization detector (FID). Aqueous samples are highly problematic because of the large vapor volume that can lead to poor peak shape and contamination of inlets, potentially causing sample carryover. Methods using aqueous samples are not well suited for use in a mass spectrometer (MS) detector system. The objective of this project was to validate a dimethyl carbonate (DMC) extraction process and GCMS method for rumen VFA analysis. To perform the extraction, 100 µL of sample, KHSO4 (500 g/L), and 2-ethylbutyrate (internal standard; 8.5 mM) were added to a microcentrifuge tube (in order) followed by 1 mL of DMC. The mixture was thoroughly vortexed and centrifuged. The organic layer (top) was removed and placed in a GC vial. The DMC extract was injected (0.5 µL) into an Agilent 5977B GCMS (8:1 split injection) with a polar DB-FFAP column. The column was held at 105 °C for 5 min, increased at 10 °C/min to 150 °C, then 65 °C/min to 240 °C, and held constant for 10 min. The peak area of acetate relative to the internal standard is linear from approximately 2 mM to at least 130 mM and encompasses the expected values of rumen concentrations for the other VFA. Recovery of VFA from spiked rumen fluid was tested at three concentrations in rumen fluid from steers fed a finishing diet or grazing wheat pasture. Recovery was not affected by the diet of the animals (P > 0.10) or the amount of VFA spiked (P > 0.19) for acetate, propionate, isobutyrate, or butyrate. There was an interaction of amount of VFA spiked and the diet of the animal (P = 0.021) for valerate and a tendency for an interaction (P = 0.051) for isovalerate, due to the recovery of the VFA being lower in the medium spike amount in rumen fluid from cattle on wheat pasture. Overall, recovery was greatest for propionate (101.9 ± 1.67%) and lowest for valerate (95.7 ± 1.95%). Including the 10-min hold at 240 °C at the end of each run prevented carryover from sample to sample. This method appears to perform well in a GCMS system and accurately and precisely quantifies rumen fluid VFA.

Scientists need to measure the end-products of microbial digestion called volatile fatty acids in the gastrointestinal tract of ruminants for many reasons. The methods currently available for the analysis of volatile fatty acids have evolved over time along with new technology. Many of the methods available are either not compatible with a mass spectrometer detector or require hazardous chemicals to prepare the samples. The method described here uses a less-hazardous chemical to prepare the samples for analysis with a mass spectrometer detector. The method tested in this manuscript was adequate in preparing samples for volatile fatty acid analysis.

Retinal Drusen Are More Common and Larger in Systemic Lupus Erythematosus With Renal Impairment.

Introduction: Complement has been implicated in systemic lupus erythematosus (SLE) pathogenesis on the basis of the associations with inherited complement defects and genome-wide association study risk alleles, glomerular deposits, reduced serum levels, and occasional reports of retinal drusen. This study examined drusen in SLE and their clinical significance. Methods: This cross-sectional observational study compared individuals with SLE recruited from renal and rheumatology clinics with hospital controls. Participants were reviewed for clinical features and underwent imaging with a nonmydriatic retinal camera. Deidentified images were examined by 2 trained graders for drusen number and size using a grid overlay. Results: The cohort with SLE (n = 65) comprised 55 women (85%) and 10 men (15%) with a median age of 47 years (interquartile range 35-59), where 23 (35%) were of southern European or Asian ancestry, and 32 (49%) had biopsy-proven lupus nephritis. Individuals with SLE had higher mean drusen numbers than controls (27 ± 60, 3 ± 9, respectively, P = 0.001), more drusen counts ≥10 (31, 48% and 3, 5%, respectively, P < 0.001), and more medium-large drusen (14, 22% and 3, 5%, respectively, P < 0.001). In SLE, mean drusen counts were higher, and drusen were larger, with an estimated glomerular filtration rate (eGFR) <90 ml/min per 1.73 m2 (P = 0.02, P = 0.02, respectively) or class IV nephritis (P = 0.03, P = 0.02). Conclusion: Drusen composition resembles that of glomerular immune deposits. CFH controls complement activation in the extracellular matrix and CFH risk variants are shared by drusen in macular degeneration and by SLE. CFH represents a possible treatment target for SLE especially with renal impairment.

Effects of supplementation rate of an extruded dried distillers' grains cube fed to growing heifers on voluntary intake and digestibility of bermudagrass hay.

Our objectives were to 1) investigate the difference in chemical composition and disappearance kinetics between loose dried distillers' grains (DDG) and extruded DDG cubes and 2) evaluate the effects of supplementation rate of extruded DDG cubes on voluntary dry matter intake (DMI), rate and extent of digestibility, and blood parameters of growing beef heifers offered ad libitum bermudagrass (Cynodon dactylon) hay. To characterize the changes in chemical composition during the extrusion process, loose and extruded DDG were evaluated via near-infrared reflectance spectroscopy, and dry matter (DM) disappearance kinetics were evaluated via time point in situ incubations. Extruded DDG cubes had greater (P ≤ 0.01) contents of fat, neutral detergent insoluble crude protein, and total digestible nutrients, but lower (P ≤ 0.01) neutral and acid detergent fiber than loose DDG. Additionally, the DM of extruded DDG cubes was more immediately soluble (P < 0.01), had greater (P < 0.01) effective degradability and lag time, and tended (P = 0.07) to have a greater disappearance rate than loose DDG. In the 29-d supplementation rate study, 23 Charolais-cross heifers were randomly assigned to one of four supplemental treatments: 1) control, no supplement; 2) low, 0.90 kg DDG cubes per d; 3) intermediate, 1.81 kg DDG cubes per d; or 4) high, 3.62 kg DDG cubes per d. Titanium dioxide was used as an external marker to estimate fecal output and particulate passage rate (Kp). Blood was collected from each animal to determine supplementation effects on blood metabolites. Indigestible neutral detergent fiber was used as an internal marker to assess the rate and extent of hay and diet DM digestibility (DMD). Increasing supplementation rate increased Kp and total diet DMI linearly (P < 0.01), yet linearly decreased (P < 0.01) hay DMI. Hay DMD decreased quadratically (P < 0.01), while total diet DMD increased linearly (P < 0.01) with increased DDG cube inclusion. Supplemented heifers had greater (P = 0.07) blood urea nitrogen concentrations than control animals 4 h post-supplementation. Intermediate and high rates of supplementation resulted in lower (P < 0.01) serum nonesterified fatty acid concentrations post-supplementation than control heifers. Concentrations of serum glucose and lactate were greatest (P ≤ 0.06) 8 h post-supplementation. Our results suggest that extruded DDG cubes may be an adequate supplement for cattle consuming moderate-quality forage, and further research is warranted.

Growing cattle are oftentimes provided supplemental concentrate as a source of protein and energy in order to meet performance goals when consuming low-quality forages. The effects of supplemental concentrate on forage intake vary, which may be related to the quality of forage and the characteristics of the supplement being evaluated. Dried distillers' grains (DDG) are a by-product of ethanol production and have become a common supplement for growing cattle due to the increased energy and rumen undegradable protein content. A stable DDG cube made via a novel extrusion process may be advantageous for pasture supplementation due to the reduced risk of loss of product from wind and soil mixing that is common with loose DDG. The effects of supplementation rate of traditional concentrate sources on forage intake are abundant, but research regarding extruded DDG cubes is almost nonexistent. Thus, our objective was to evaluate extruded DDG cube supplementation rate (0, 0.90, 1.81, or 3.62 kg DDG cubes per d) for growing cattle on voluntary intake and digestibility of moderate-quality forage. Although increasing supplementation rate reduced forage intake and digestibility, total diet intake and digestibility were increased. Our results suggested that extruded DDG cubes have potential as a supplement for cattle consuming moderate-quality forage.