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The microbiome is known to play a role in many human diseases, but identifying key microbes and their functions generally requires large studies due to the vast number of species and genes, and the high levels of intra-individual and inter-individual variation. 16S amplicon sequencing of the rRNA gene is commonly used for large studies due to its comparatively low sequencing cost, but it has poor taxonomic and functional resolution. Deep shotgun sequencing is a more accurate and comprehensive alternative for small studies, but can be cost-prohibitive for biomarker discovery in large populations. Shallow or moderate-depth shotgun metagenomics may serve as a viable alternative to 16S sequencing for large-scale and/or dense longitudinal studies, but only if resolution and reproducibility are comparable. Here we applied both 16S and shallow shotgun stool microbiome sequencing to a cohort of 5 subjects sampled twice daily and weekly, with technical replication at the DNA extraction and the library preparation/sequencing steps, for a total of 80 16S samples and 80 shallow shotgun sequencing samples. We found that shallow shotgun sequencing produced lower technical variation and higher taxonomic resolution than 16S sequencing, at a much lower cost than deep shotgun sequencing. These findings suggest that shallow shotgun sequencing provides a more specific and more reproducible alternative to 16S sequencing for large-scale microbiome studies where costs prohibit deep shotgun sequencing and where bacterial species are expected to have good coverage in whole-genome reference databases., (© 2023. The Author(s).)

Introduction: Predicting length of stay (LOS) is beneficial to patients and the health service. When a prolonged LOS is predicted, it gives the opportunity for focused therapies and allocation of resources to reduce this period. In emergency general surgery (EGS) there has been limited investigation of variables that may be important predictors of LOS. This study examines social characteristics alongside measures of severity of acute illness and co-morbidities in an adult EGS population to establish their contribution to LOS., Methods: Data were collected prospectively from patients at admission including medical variables, demographics, and therapeutic requirements. The length of hospital admission was measured, and multiple regression analysis was used to identify variables which predicted the LOS., Results: Data were collected from 105 patients. The regression model gave an R 2 of 0.34, p  = 0.0006. Barthal index (measure of independence in activities of daily living) was a significant predictor of LOS [logworth 1.649, p0.02243 ]. Housing status and Level of social support both correlated in one-way analysis with LOS., Conclusion: There are non-surgical variables, measurable at admission which are of significant value in predicting LOS of EGS patients. This warrants further investigation through a larger study to better quantify the contributions of these variables, and establish potential early interventions to reduce the LOS., Competing Interests: None., (Crown Copyright © 2021 Published by Elsevier Ltd on behalf of IJS Publishing Group Ltd.)

Background: The gut microbiome harbors trillions of bacteria that play a major role in dietary nutrient extraction and host metabolism. Metabolic diseases such as obesity and diabetes are associated with shifts in microbiome composition and have been on the rise in Westernized or highly industrialized countries. At the same time, Westernized diets low in dietary fiber have been shown to cause loss of gut microbial diversity. However, the link between microbiome composition, loss of dietary fiber, and obesity has not been well defined., Results: To study the interactions between gut microbiota, dietary fiber, and weight gain, we transplanted captive and wild douc gut microbiota into germ-free mice and then exposed them to either a high- or low-fiber diet. The group receiving captive douc microbiota gained significantly more weight, regardless of diet, while mice receiving a high-fiber diet and wild douc microbiota remained lean. In the presence of a low-fiber diet, the wild douc microbiota partially prevented weight gain. Using 16S rRNA gene amplicon sequencing we identified key bacterial taxa in each group, specifically a high relative abundance of Bacteroides and Akkermansia in captive douc FMT mice and a higher relative abundance of Lactobacillus and Clostridium in the wild douc FMT mice., Conclusions: In the context of our germ-free mouse experiment, wild douc microbiota could serve as a reservoir for microbes for cross-species transplants. Our results suggest that wild douc microbiota are tailored to diverse fiber diets and can prevent weight gain when exposed to a native diet.

The microbiome is important to all animals, including poultry, playing a critical role in health and performance. Low-dose antibiotics have historically been used to modulate food production animals and their microbiome. Identifying alternatives to antibiotics conferring similar modulatory properties has been elusive. The purpose of this study was to determine if a host-tailored probiotic could recapitulate effects of a low-dose antibiotic on host response and the developing microbiome. Over 13 days of life, turkey poults were supplemented continuously with a low-dose antibiotic or oral supplementation of a prebiotic with or without two different probiotics (8 cage units, n  = 80 per group). Gastrointestinal bacterial and fungal communities of poults were characterized by 16S rRNA gene and ITS2 amplicon sequencing. Localized and systemic host gene expression was assessed using transcriptome sequencing (RNA-Seq), kinase activity was assessed by avian-specific kinome peptide arrays, and performance parameters were assessed. We found that development of the early-life microbiome of turkey poults was tightly ordered in a tissue- and time-specific manner. Low-dose antibiotic and turkey-tailored probiotic supplementation, but not nontailored probiotic supplementation, elicited similar shifts in overall microbiome composition during development compared to controls. Treatment-induced bacterial changes were accompanied by parallel shifts in the fungal community and host gene expression and enhanced performance metrics. These results were validated in pen trials that identified further additive effects of the turkey-tailored probiotic combined with different prebiotics. Alternative approaches to low-dose antibiotic use in poultry are feasible and can be optimized utilizing the indigenous poultry microbiome. Similar approaches may also be beneficial for humans. IMPORTANCE Alternative approaches are greatly needed to reduce the need for antibiotic use in food animal production. This study utilized a pipeline for the development of a host-tailored probiotic to enhance performance in commercial turkeys and modulate their microbiota, similar to the effects of low-dose antibiotic administration. We determined that a host-tailored probiotic, developed in the context of the commercial turkey gut microbiome, was more effective at modulating these parameters than a nontailored probiotic cocktail. Furthermore, the host-tailored probiotic mimicked many of the effects of a low-dose antibiotic growth promoter. Surprisingly, the effects of the antibiotic growth promoter and host-tailored probiotic were observed across kingdoms, illustrating the coordinated interkingdom effects of these approaches. This work suggests that tailored approaches to probiotic development hold promise for modulating the avian host and its microbiota., (Copyright © 2019 Ward et al.)

Diet is a key determinant of human gut microbiome variation. However, the fine-scale relationships between daily food choices and human gut microbiome composition remain unexplored. Here, we used multivariate methods to integrate 24-h food records and fecal shotgun metagenomes from 34 healthy human subjects collected daily over 17 days. Microbiome composition depended on multiple days of dietary history and was more strongly associated with food choices than with conventional nutrient profiles, and daily microbial responses to diet were highly personalized. Data from two subjects consuming only meal replacement beverages suggest that a monotonous diet does not induce microbiome stability in humans, and instead, overall dietary diversity associates with microbiome stability. Our work provides key methodological insights for future diet-microbiome studies and suggests that food-based interventions seeking to modulate the gut microbiota may need to be tailored to the individual microbiome. Trial Registration: ClinicalTrials.gov: NCT03610477., (Copyright © 2019 Elsevier Inc. All rights reserved.)

Many US immigrant populations develop metabolic diseases post immigration, but the causes are not well understood. Although the microbiome plays a role in metabolic disease, there have been no studies measuring the effects of US immigration on the gut microbiome. We collected stool, dietary recalls, and anthropometrics from 514 Hmong and Karen individuals living in Thailand and the United States, including first- and second-generation immigrants and 19 Karen individuals sampled before and after immigration, as well as from 36 US-born European American individuals. Using 16S and deep shotgun metagenomic DNA sequencing, we found that migration from a non-Western country to the United States is associated with immediate loss of gut microbiome diversity and function in which US-associated strains and functions displace native strains and functions. These effects increase with duration of US residence and are compounded by obesity and across generations., (Copyright © 2018 Elsevier Inc. All rights reserved.)

Objective: The objective of the study was to investigate the effects of zinc amino acid complex (ZnAA) on growth performance, hematological and biochemical parameters in weanling pigs., Methods: In Exp. 1, a total of 216 Duroc×Landrace×Large White weanling pigs were assigned randomly to 6 dietary treatments. Each treatment had 6 replicates (pens) with 6 pigs each. The diets were corn-soybean meal based with supplementation of 0, 20, 40, 80, 120 mg Zn/kg from ZnAA or 40 mg Zn/kg from feed-grade zinc sulfate. The experiment lasted 42 days. In Exp. 2, a total of 180 weanling pigs were assigned randomly to 3 dietary treatments supplemented with 0, 80, or 800 mg Zn/kg from ZnAA., Results: In Exp. 1, pigs fed 40 to 80 mg Zn/kg from ZnAA had higher (p<0.05) average daily gain (ADG) than the unsupplemented group during d 0 to 14. During d 0 to 42, the pigs fed 20 to 120 mg Zn/kg from ZnAA had increased (p<0.05) ADG. Pigs fed 20 to 120 mg/kg Zn from ZnAA had lower feed:gain (p<0.05), increased the activity of serum Cu-Zn superoxide dismutase on d 14, and increased serum Zn levels on d 42 (p<0.05). In Exp. 2, pigs fed diets with 800 mg Zn/kg had increased average daily feed intake during d 15 to 28 (p<0.05) compared to the unsupplemented group. During d 0 to 28, the pigs fed supplemental Zn had increased ADG (p<0.05). On d 14 and d 28, pigs fed supplemental Zn had higher the serum alkaline phosphatase activities (p<0.05). No significant differences were observed in the hematological parameters and organ indices., Conclusion: Supplementation with 20 to 80 mg/kg Zn from ZnAA improved the growth performance in weaned pigs. The piglets can tolerate up to 800 mg/kg Zn from ZnAA with limited potential health effects.

With the advent of next-generation sequencing and microbial community characterization, we are beginning to understand the key factors that shape early-life microbial colonization and associated health outcomes. Studies characterizing infant microbial colonization have focused mostly on bacteria in the microbiome and have largely neglected fungi (the mycobiome), despite their relevance to mucosal infections in healthy infants. In this pilot study, we characterized the skin, oral, and anal mycobiomes of infants over the first month of life ( n = 17) and the anal and vaginal mycobiomes of mothers ( n = 16) by internal transcribed spacer 2 (ITS2) amplicon sequencing. We found that infant mycobiomes differed by body site, with the infant mycobiomes at the anal sites being different from those at the skin and oral sites. The relative abundances of body site-specific taxa differed by birth mode, with significantly more Candida albicans fungi present on the skin of vaginally born infants on day 30 and significantly more Candida orthopsilosis fungi present in the oral cavity of caesarean section-born infants throughout the first month of life. We found the mycobiomes within individual infants to be variable over the first month of life, and vaginal birth did not result in infant mycobiomes that were more similar to the mother's vaginal mycobiome. Therefore, although vertical transmission of specific fungal isolates from mother to infant has been reported, it is likely that other sources (environment, other caregivers) also contribute to early-life mycobiome establishment. Thus, future longitudinal studies of mycobiome and bacterial microbiome codevelopment, with dense sampling from birth to beyond the first month of life, are warranted. IMPORTANCE Humans are colonized by diverse fungi (mycobiome), which have received much less study to date than colonizing bacteria. We know very little about the succession of fungal colonization in early life and whether it may relate to long-term health. To better understand fungal colonization and its sources, we studied the skin, oral, and anal mycobiomes of healthy term infants and the vaginal and anal mycobiomes of their mothers. Generally, infants were colonized by few fungal taxa, and fungal alpha diversity did not increase over the first month of life. There was no clear community maturation over the first month of life, regardless of body site. Key body-site-specific taxa, but not overall fungal community structures, were impacted by birth mode. Thus, additional studies to characterize mycobiome acquisition and succession throughout early life are needed to form a foundation for research into the relationship between mycobiome development and human disease.

This experiment was conducted to evaluate the effects of chromium methionine with/without zinc sulfate or zinc amino acid complex on the growth performance, carcass traits, meat quality, serum parameters, endocrine parameters, and antioxidant status of growing-finishing pigs. A total of 180 (32.0 ± 1.7 kg body weight, BW) crossbred pigs (Duroc × Landrace × Yorkshire) were used in a completely randomized design with three dietary treatments and 10 replicates per treatment (five pens of barrows and five pens of gilts with six pigs per replicate). Three treatments were corn-soybean meal-based diets supplemented with 100 mg Zn/kg from zinc sulfate (ZnSO 4 ), 100 mg Zn/kg from ZnSO 4  + 0.2 mg Cr/kg from chromium methionine complex (CrMet), or 50 mg Zn/kg from ZnSO 4  + 50 mg Zn/kg from zinc amino acid complex (ZnAA) + 0.2 mg Cr/kg from CrMet, respectively. The experiment lasted 105 days, of which was divided into three stages including phase 1 (30 to 50 kg BW), phase 2 (50 to 80 kg BW), and phase 3 (80 to 110 kg BW). Results showed that supplementation with CrMet and ZnAA improved (P < 0.05) the feed conversion of the pigs in phase 2, phase 3, and the overall experiment. Hot carcass weight, dressing percentage, and a longissimus dorsi muscle area were increased (P < 0.05) in pigs fed with diets supplemented with both CrMet and ZnAA compared with pigs fed with diets containing only ZnSO 4 (P < 0.05). There was also an increase (P < 0.01) pH 24 h in the longissimus dorsi muscle in pigs fed with diets supplemented with CrMet and ZnAA. The concentration of serum glucose in pigs fed with diets containing CrMet and ZnAA was decreased (P < 0.05) compared with that in pigs fed with the diet containing ZnSO 4 . Supplementation with CrMet and ZnAA increased (P < 0.05) the circulating levels of insulin and decreased (P < 0.05) cortisol. There was an increase (P < 0.05) in total serum antioxidant capacity and Cu/Zn superoxide dismutase activity as well as a decrease (P < 0.05) in serum malondialdehyde concentrations in pigs fed with diets supplemented with CrMet and ZnAA compared with pigs fed with the diet supplemented only with ZnSO 4 . In conclusion, supplementation of CrMet only or CrMet together with ZnAA improved feed conversion, carcass traits, and meat quality in the growing-finishing pigs.

The microbes colonizing the infant gastrointestinal tract have been implicated in later-life disease states such as allergies and obesity. Recently, the medical research community has begun to realize that very early colonization events may be most impactful on future health, with the presence of key taxa required for proper immune and metabolic development. However, most studies to date have focused on bacterial colonization events and have left out fungi, a clinically important sub-population of the microbiota. A number of recent findings indicate the importance of host-associated fungi (the mycobiota) in adult and infant disease states, including acute infections, allergies, and metabolism, making characterization of early human mycobiota an important frontier of medical research. This review summarizes the current state of knowledge with a focus on factors influencing infant mycobiota development and associations between early fungal exposures and health outcomes. We also propose next steps for infant fungal mycobiome research, including longitudinal studies of mother-infant pairs while monitoring long-term health outcomes, further exploration of bacterium-fungus interactions, and improved methods and databases for mycobiome quantitation.

The digestive function of the stomach depends on acidification of the gastric lumen. Acid secretion into the lumen is triggered by activation of the PKA cascade, which ultimately results in the insertion of gastric H,K-ATPases into the apical plasma membranes of parietal cells. A coupling protein is ezrin, whose phosphorylation at Ser-66 by PKA is required for parietal cell activation. However, little is known regarding the molecular mechanism(s) by which this signaling pathway operates in gastric acid secretion. Here we show that PKA cooperates with MST4 to orchestrate histamine-elicited acid secretion by phosphorylating ezrin at Ser-66 and Thr-567. Histamine stimulation activates PKA, which phosphorylates MST4 at Thr-178 and then promotes MST4 kinase activity. Interestingly, activated MST4 then phosphorylates ezrin prephosphorylated by PKA. Importantly, MST4 is important for acid secretion in parietal cells because either suppression of MST4 or overexpression of non-phosphorylatable MST4 prevents the apical membrane reorganization and proton pump translocation elicited by histamine stimulation. In addition, overexpressing MST4 phosphorylation-deficient ezrin results in an inhibition of gastric acid secretion. Taken together, these results define a novel molecular mechanism linking the PKA-MST4-ezrin signaling cascade to polarized epithelial secretion in gastric parietal cells., (© 2015 by The American Society for Biochemistry and Molecular Biology, Inc.)

Classification of the human gut microbiome into distinct types, or "enterotypes," provides an attractive framework for understanding microbial variation in health and disease. However, as discussed here, several different methods of collapsing enterotype variation into a few discrete clusters suggest that enterotype distribution is continuous and can vary widely within an individual., (Copyright © 2014 Elsevier Inc. All rights reserved.)

Soluble CD14 (sCD14) is a pattern recognition receptor and Toll-like co-receptor observed in human milk (5-26μg/mL) and other bodily fluids such as blood (3μg/mL). The most well defined role of sCD14 is to recognize lipopolysaccharide of Gram-negative bacteria and signal an immune response through Toll-like receptor 4 (TLR4). Previous research has shown ingested sCD14 to transfer from the gastrointestinal tract and into the blood stream in neonatal rats. The contribution of human milk sCD14 to circulating levels in the infant and the functionality of the protein, however, remained unknown. Using CD14(-/-) mouse pups fostered to wild type (WT) mothers expressing sCD14 in their milk, we show herein that ingestion of sCD14 resulted in blood sCD14 levels up 0.16±0.09μg/mL. This represents almost one-third (26.7%) of the circulating sCD14 observed in WT pups fostered to WT mothers (0.60±0.14μg/mL). We also demonstrate that ingested-sCD14 transferred to the blood remains functional in its ability to recognize lipopolysaccharide as demonstrated by a significant increase in immune response (IL-6 and TNF-α) in CD14(-/-) pups fostered to WT mothers in comparison to control animals (P=0.002 and P=0.007, respectively). Using human intestinal cells (Caco-2), we also observed a significant decrease in sCD14 transcytosis when TLR4 was knocked down (P<0.001), suggesting sCD14 transfer involves TLR4. The bioavailability of human milk sCD14 established in this report confirms the importance of human milk proteins for the infant and demonstrates the need to improve infant formulas which are lacking in immune proteins such as sCD14., (Copyright © 2014 The Authors. Published by Elsevier GmbH.. All rights reserved.)

Background: Milk acts as an edible immune system that is transferred from mother to newborn. Soluble Cluster of Differentiation 14 (sCD14) is a protein found in significant quantities in human milk (~8-29 µg/ml). At a 10-fold lower concentration in the blood (~3 µg/ml), the most notable role of sCD14 is to sequester lipopolysaccharides of Gram-negative bacteria from immune cells., Methods: To explore the pharmacodynamics of this milk protein and its biological fate, the biodistribution of radiolabeled sCD14 ((14)C, (125)I) was monitored in 10-d-old rat pups., Results: Up to 3.4 ± 2.2% of the radiolabeled sCD14 administered was observed, intact, in the pup blood for up to 8 h post-ingestion. Additionally, 30.3 ± 13.0% of the radiolabeled sCD14 administered was observed degraded in the stomach at 8 h post-ingestion. A reservoir of intact, administered sCD14 (3.2 ± 0.3%), however, remained in the stomach at 8 h post-ingestion. Intact sCD14 was observed in the small intestine at 5.5 ± 1.6% of the dose fed at 8 h post-ingestion., Conclusion: The presence of intact sCD14 in the blood and the gastrointestinal tract of newborns post-ingestion has implications in the development of allergies, obesity, and other inflammation-related pathogeneses later in life.

Background: Human milk contains a diverse population of bacteria that likely influences colonization of the infant gastrointestinal tract. Recent studies, however, have been limited to characterization of this microbial community by 16S rRNA analysis. In the present study, a metagenomic approach using Illumina sequencing of a pooled milk sample (ten donors) was employed to determine the genera of bacteria and the types of bacterial open reading frames in human milk that may influence bacterial establishment and stability in this primal food matrix. The human milk metagenome was also compared to that of breast-fed and formula-fed infants' feces (n = 5, each) and mothers' feces (n = 3) at the phylum level and at a functional level using open reading frame abundance. Additionally, immune-modulatory bacterial-DNA motifs were also searched for within human milk., Results: The bacterial community in human milk contained over 360 prokaryotic genera, with sequences aligning predominantly to the phyla of Proteobacteria (65%) and Firmicutes (34%), and the genera of Pseudomonas (61.1%), Staphylococcus (33.4%) and Streptococcus (0.5%). From assembled human milk-derived contigs, 30,128 open reading frames were annotated and assigned to functional categories. When compared to the metagenome of infants' and mothers' feces, the human milk metagenome was less diverse at the phylum level, and contained more open reading frames associated with nitrogen metabolism, membrane transport and stress response (P < 0.05). The human milk metagenome also contained a similar occurrence of immune-modulatory DNA motifs to that of infants' and mothers' fecal metagenomes., Conclusions: Our results further expand the complexity of the human milk metagenome and enforce the benefits of human milk ingestion on the microbial colonization of the infant gut and immunity. Discovery of immune-modulatory motifs in the metagenome of human milk indicates more exhaustive analyses of the functionality of the human milk metagenome are warranted.

Mortality in swine herds is often associated with lameness, and trace minerals are implicated in maintaining integrity of skeletal tissues. The objectives of this study were to determine if prolific sows displayed evidence of trace mineral depletion with age and to determine the prevalence of osteochondrosis (OC) lesions. Reduced mineral concentrations with age would support recommendations for an increase in the amount of dietary minerals. Tissue samples were collected from 66 sows selected to represent a cross-sectional profile of a prolific herd fed diets with inorganic sources of trace minerals fortified at concentrations typically found in commercial diets. Females ranged from nulliparous (parity 0) to parity 7 with a lifetime average of 12.9 ± 0.5 pigs born alive per litter. Minerals were assessed in humerus, scapula, ovary, liver, and muscle (psoas major) tissues. Percent bone ash increased (P < 0.05) with parity from 64 to 66% but differed among bone sections. The Ca (39.0%) and P (18.9%) concentrations in bone ash were essentially constant in all sections and parities. Bone Cu, Fe, Mn, and Zn concentrations varied among sections, but differences due to parity (P < 0.05) were only detected in Fe. Bone Fe decreased from approximately 49 μg/g ash in parity 0 and 1 sows to approximately 29 μg/g ash in parity 7, likely reflecting loss of hemopoietic tissue with age. No evidence was detected in liver for depletion of trace minerals across parity; however, liver Cu and Zn concentrations tended to increase with age. Liver Mn concentrations varied with parity, but no consistent trend with parity was evident. Ovary Cu and Mn concentrations varied dramatically as a function of the reproductive status, but no evidence was detected for depletion with parity. Articular surfaces of the distal scapula and proximal and distal humerus were evaluated grossly for prevalence of OC; bones were then sectioned to evaluate lesions in subchondral bone and physis. Incidence of OC lesions on the articular-epiphyseal cartilage complex varied among bone sites, but differences across parities were not detected. In a subset of sows with subchondral bone lesions, the lesions appeared severe enough to contribute to clinical lameness, particularly in the distal humerus site. However, none of the sows exhibited lameness at slaughter. As no reductions in mineral concentrations with age were detected, recommendations to increase dietary mineral supplementation with age were not supported.

Sources of Zn, Mn, and Cu (IZMC) as sulfates or as amino acid complexes (OZMC) were used to supplement Cobb 500 breeder hen diets. Experimental treatments consisted of diets supplemented with 1) 100, 100, and 10 mg/kg of Zn, Mn, and Cu, respectively, from IZMC (control); 2) 60, 60, and 3 mg/kg of Zn, Mn, and Cu, respectively, from IZMC plus 40, 40, and 7 mg/kg of Zn, Mn, and Cu, respectively, from OZMC (ISO); and 3) a diet with 100, 100, and 10 mg/kg of Zn, Mn, and Cu, respectively, from IZMC as in control plus 40, 40, and 7 mg/kg of supplemental Zn, Mn, and Cu from OZMC (on top). Ten replications of 20 females and 2 males were used per treatment. Eggs from breeders at 30, 40, 50 and 60 wk of age were incubated, and 5 embryos per replicate were collected at 10 (E10), 14 (E14), and 18 (E18) d of incubation. Midshaft width and calcification were measured for left tibia and femur stained with Alcian Blue and Alizarin Red S. At hatch, the left tibia of 5 chicks per replicate was sampled for histological evaluation of the diaphysis and distal epiphysis. Feeding the ISO treatment compared with the control diet increased the Zn (P < 0.05) but not Mn and Cu content of the yolk and albumen blend. At E14, the ISO and on-top treatments had a trend to increase tibia calcification at the rates of 1.6 and 1%, respectively (P < 0.1). The E18 ISO and on-top treatments had 2% thicker tibia compared with the control, regardless of hen age (P < 0.05). Also, at E18, calcification of tibia and femur was higher from hens fed the on-top treatment (P < 0.05). The chicks from the ISO and on-top groups had increased tibia moment of inertia (P < 0.01) at day of hatch. Broiler breeder hens consuming OZMC associated with IZMC produced embryos and hatching chicks with improvements in selected bone mineralization parameters.

The objective of the current study was to determine the bioavailability of an organic zinc source (Availa-Zn) compared with zinc sulfate in a European-type broiler diet. A total of 480 one-day-old male Ross 308 broilers were housed in 48 digestibility cages (10 birds per cage), being randomly divided over 9 treatments. At d 3, the number of birds was standardized to 8. Birds were fed a basal wheat-maize-soya diet (containing 33.5 mg of Zn/kg) with different supplementation levels of zinc (reference zinc source: inorganic zinc sulfate: 0, 5, 10, 15, 20, and 40 mg of Zn/kg of feed; test zinc source: Availa-Zn: 0, 5, 10, 15 mg of Zn/kg of feed). Production performance and tibia zinc content were measured. There were no differences in production performance between the different zinc sources when fed to broilers until 21 d of age. Tibia zinc content was increased linearly with the dietary zinc content up to 20 mg/kg zinc sulfate. The relative biological value of organic zinc was 1.64 compared with zinc sulfate as a reference zinc source (1.00), as indicated by the slope ratio of the linear response curves for both zinc sources, using tibia zinc content as a response parameter. In a practical European broiler diet, the organic Availa-Zn had a higher bioavailability than inorganic zinc sulfate.

Nitrous oxide (N(2)O), the third most abundant greenhouse gas (GHG), is highly stable and plays a significant role in stratospheric ozone destruction. The primary anthropogenic source of N(2)O stems from use of nitrogen fertilizers in soil. The bacterial enzyme nitrous oxide reductase (N(2)OR), naturally found in some soils, is the only known enzyme capable of catalyzing the final step of the denitrification pathway, conversion of N(2)O to N(2). In this opinion, we discuss potential biology-based strategies to reduce N(2)O by amplifying the amount of available enzyme catalyst in agri-system environments during crop growth and in post-harvest detritus. N(2)OR from Pseudomonas stutzeri has been tested in transgenic plants with promising results. Such seed-borne phytoremediation systems targeted towards GHGs merit field testing., (Copyright © 2012. Published by Elsevier Ltd.)

The economical preparation of microgram quantities of (14)C-labeled proteins by in vacuo methylation with methyl iodide is described. The (14)C radiolabeling was achieved by the covalent attachment of [(14)C]methyl groups onto amino and imidazole groups by reaction in vacuo with [(14)C]methyl iodide. The method was tested by investigating the biodistribution of (14)C in rats that were fed (14)C-labeled human soluble cluster of differentiation 14 (CD14) protein, a receptor for bacterial lipopolysaccharide. Two other control proteins, bovine serum albumin (BSA) and casein, were also labeled with (14)C and used for comparative analysis to determine the following: (i) the efficacy and cost efficiency of the in vacuo radiolabeling procedure and (ii) the extent of incorporation of the (14)C label into the organs of orogastrically fed 10-day-old Sprague-Dawley rats. [(14)C]BSA, [(14)C]casein, and [(14)C]CD14 were individually prepared with specific radioactivities of 34,400, 18,800, and 163,000 disintegrations per minute (dpm)/microg, respectively. It was found that the accumulation of (14)C label in the organs of [(14)C]CD14-fed rats, most notably the persistence of (14)C in the stomach 480 min postgavage, was temporally and spatially distinct from [(14)C]BSA and [(14)C]casein-fed rats., (2010 Elsevier Inc. All rights reserved.)

Four experiments were conducted to determine the interactive effects of pharmacological amounts of Zn from ZnO and Cu from organic (Cu-AA complex; Cu-AA) or inorganic (CuSO(4)) sources on growth performance of weanling pigs. The Cu was fed for 4 (Exp. 1) or 6 (Exp. 2, 3, and 4) wk after weaning, and Zn was fed for 4 (Exp. 1) or 2 (Exp. 2, 3, and 4) wk after weaning. Treatments were replicated with 7 pens of 5 or 6 pigs per pen (19.0 ± 1.4 d of age and 5.8 ± 0.4 kg of BW, Exp. 1), 12 pens of 21 pigs per pen (about 21 d of age and 5.3 kg of BW, Exp. 2), 5 pens of 4 pigs per pen (20.3 ± 0.5 d of age and 7.0 ± 0.5 kg of BW, Exp. 3), and 16 pens of 21 pigs per pen (about 21 d of age and 5.7 kg of BW, Exp. 4). In Exp. 1 and 2, Cu-AA (0 vs. 100 mg/kg of Cu) and ZnO (0 vs. 3,000 mg/kg of Zn) were used in a 2 × 2 factorial arrangement. Only Exp. 1 used in-feed antibiotic (165 mg of oxytetracycline and 116 mg of neomycin per kilogram feed), and Exp. 2 was conducted at a commercial farm. In Exp. 3, sources of Cu (none; CuSO(4) at 250 mg/kg of Cu; and Cu-AA at 100 mg/kg of Cu) and ZnO (0 vs. 3,000 mg/kg of Zn) were used in a 3 × 2 factorial arrangement. In Exp. 4, treatments were no additional Cu, CuSO(4) at 315 mg/kg of Cu, or Cu-AA at 100 mg/kg of Cu to a diet supplemented with 3,000 mg/kg of Zn from ZnO and in-feed antibiotic (55 mg of carbadox per kilogram of feed). In Exp. 1 and 2, both Zn and Cu-AA improved (P < 0.001 to P = 0.03) ADG and ADFI. No interactions were observed, except in wk 1 of Exp. 2, where Zn increased the G:F only in the absence of Cu-AA (Cu-AA × Zn, P = 0.04). A naturally occurring colibacillosis diarrhea outbreak occurred during this experiment. The ZnO addition reduced (P < 0.001) the number of pigs removed and pig-days on antibiotic therapy. In Exp 3, ADFI in wk 2 was improved by Zn and Cu (P < 0.001 and P = 0.09, respectively) with no interactions. In wk 1, G:F was reduced by ZnO only in the absence of Cu (Cu × Zn, P = 0.03). Feeding Zn decreased fecal microbiota diversity in the presence of CuSO(4) but increased it in the presence of Cu-AA (Cu source × Zn, P = 0.06). In Exp. 4, Cu supplementation improved the overall ADG (P = 0.002) and G:F (P < 0.001). The CuSO(4) effect on G:F was greater (P < 0.001) than the Cu-AA effect. Our results indicate that pharmacological amounts of ZnO and Cu (Cu-AA or CuSO(4)) are additive in promoting growth of pigs after weaning.

Mother's milk represents a foundational step in the proper development of newborn immunity. This is achieved, in part, through the action of numerous regulatory proteins such as soluble cluster of differentiation 14 (sCD14) found in significant quantities in human milk (~25-50 μg/mL). In adults, CD14 stimulates cytokine production in response to lipopolysaccharide (LPS), the major lipid component found in the outer membrane of Gram-negative bacteria. However, the fate and function of sCD14 in the neonatal gastrointestinal (GI) tract are unknown and may function differently from adults. Therefore, we administered human sCD14 to experimental animals and observed that it persisted in the upper GI tract after feeding. In our search for potential proteolytic protectants, immunoprecipitation of sCD14 from human milk revealed a 15-kD novel protein that copurified with sCD14. Mass spectrometry analysis of the protein identified alpha-lactalbumin. CD14 was also identified by immunoblot after immunoprecipitation of alpha-lactalbumin from milk. In vitro digestion assays revealed that purified alpha-lactalbumin decreases the proteolytic degradation of human milk derived sCD14 in vitro, suggesting a mechanism by which this key LPS receptor may remain functional in the neonate gut.

Hollow spherical nanoparticles with ordered mesoporous silica shells were fabricated by evaporation-induced self-assembly using (NH(4))(2)SO(4) and cetyltrimethyl ammonium bromide (CTAB) as templates. The model drug, L-methionine, was encapsulated within the spherical void at high loadings by repeated crystallization.

Transgenic mouse mutation detection systems allow investigation of the origins and mechanisms of mutation associated with exogenous and endogenous mutagen exposures in individual tissues and cell types. In the past, selection assays for transgenic mutants have been contaminated with nonmurine-derived mutations and assay validation is critical to ensure murine in vivo origins of mutations. This is critical in studies of spontaneous mutations and extrapolation to endogenous mammalian genes. Herein, we provide one measure of the contribution of Escherichia coli (E. coli)-derived mutations to the Big Blue(R) cII transgene mutant selection assay. We report the first direct evidence of an E. coli-derived cII mutation identified among mutations recovered in the cII selective assay. An E. coli transposable 5 (Tn5) element IS50R inverted repeat (1,534 bp) was identified at base pair 414 in the cII transgene and the insertion generated a 9 bp target site duplication typical of this type of transposition. The bacterial transposition occurred only once in the assay of 25 x 10(6) plaque forming units and sequencing of 1,177 cII mutants. The observed frequency of this type of mutation is 4 x 10(-8) in retrieved lambda phage and 8.5 x 10(-4) in harvested cII mutants and thus a very rare occurrence in typical analyses of spontaneous in vivo mutations. Given that the frequency of transposition is equal to, or an order of magnitude higher, than the frequency of point mutations in E. coli, this article provides excellent validation for the murine origins of mutations detected using the cII mutant selection assay.

Sixty-four pigs from 16 sows were used to evaluate addition of zinc amino acid complex (ZnAA) to lactating sows and gastric nutriment-intubation of zinc methionine (ZnMet) to suckling pigs on mineral status, intestinal morphology and bacterial translocation after weaning. Sows were fed a barley-based diet supplying 120 ppm zinc (Zn; control) or the control diet supplemented with 240 ppm Zn from ZnAA. At birth, day-10 and day-21 (weaning) of age, pigs from each litter were nutriment-intubated with 5 ml of an electrolyte solution without or with 40 mg Zn from ZnMet. At weaning, 24 h prior to the collection of small and large intestinal lymph nodes and sections of the duodenum, jejunum and ileum, the pigs received an intramuscular injection of saline without or with 150 microg/kg body weight of Escherichia coli O26:B6 lipopolysaccharide (LPS). With the exception of a tendency (p = 0.09) for lower serum concentration of copper in pigs at weaning from ZnAA-supplemented sows, there were no differences (p > 0.1) than for pigs from control-fed sows for mineral status or intestinal morphology. Nutriment-intubation of ZnMet increased serum (p = 0.001) and liver (p = 0.003) Zn concentrations, number of goblet cells per 250 microm length of jejunal villous epithelium (p = 0.001) and tended (p = 0.06) to enhance jejunum mucosa thickness. Interactive effects (p < 0.05) for higher jejunal villi height and villi:crypt ratio and increased ileal goblet cell counts were apparent for pigs from ZnAA-supplemented sows that also received nutriment-intubation of ZnMet. Challenge with LPS increased (p = 0.05) ileal villous width. Nutriment-intubation of ZnMet decreased (p = 0.05) anaerobic bacteria colony forming unit counts in the large intestinal mesenteric lymph nodes. In conclusion, nutriment-intubation of ZnMet increased serum and liver tissue concentrations of Zn and resulted in limited improvement to intestinal morphology of weaned pigs.

The evolving composition of evaporating ethanol-water droplets (initially 32.6 or 45.3 microm radius) is probed by stimulated Raman scattering over the period 0.2 to 3 ms following droplet generation and with a surrounding nitrogen gas pressure in the range 10 to 100 kPa. The dependence of the evaporation rate on the relative humidity of the surrounding gas phase is also reported. The measured data are compared with both a quasi-steady state model and with numerical simulations of the evaporation process. Results from the numerical simulations are shown to agree closely with the measurements when the stimulated signal is assumed to arise from an outer shell with a probe depth of 2.9+/-0.4% of the droplet radius, consistent with a previous determination. Further, the time-dependent measurements are shown to be sensitive to the development of concentration gradients within evaporating droplets. This represents the first direct measurement of the spatial gradients in composition that arise during the evaporation of aerosol droplets and allows the influence of liquid phase diffusion within the condensed phase on droplet evaporation to be examined.

Eighty male pigs from 20 litters were used to evaluate dietary addition of 250mg/kg of Zn from zinc amino acid complex (ZnAA) to sows during the last trimester of gestation and gastric intubation of 40mg Zn from soluble zinc methionine (ZnMet) to suckling pigs at birth and on day 7 and 14 (weaning) on small intestinal morphology, Zn status and bacterial translocation in early-weaned pigs. At weaning, pigs were challenged with an intramuscular injection of saline without or with 120microg/kg BW of lipopolysaccharide (LPS; from Escherichia coli O26:B6) and were euthanized 24h later prior to collection of intestinal lymph nodes and small intestinal sections. Zinc concentration in serum 7 days after birth and at weaning were higher in pigs from ZnAA-supplemented sows and those receiving gastric intubation with ZnMet (P=0.05 and P<0.0001, respectively). Post-weaning liver tissue concentrations for Zn (P<0.0001) and Fe (P=0.04) were higher and for Cu lower (P<0.0001) in pigs intubated with ZnMet. Pigs from ZnAA-supplemented compared with control-fed sows tended (P<0.1) to have increased villi height and villus:crypt ratio in the jejunum and higher (P=0.1) goblet cell counts in the ileum. Goblet cell counts of ZnMet-intubated (P=0.03) and LPS-challenged pigs (P=0.05) were also higher in the jejunum. Supplementation of ZnAA to gestating sows increased (P=0.04) E. coli colony forming unit counts in the small intestinal mesenteric lymph nodes of early-weaned pigs. In conclusion, dietary addition of ZnAA to gestating sows and gastric intubation of ZnMet improved Zn status of suckling pigs.

Ordered mesoporous carbon/silica nanocomposites were synthesized through a novel multi-component molecular assembly and show promising potential as corrosion-resisted electrocatalyst supports.

Thermosensitive inorganic-organic hybrid polymers and gels can be used for controlled molecular transport in a variety of applications that require robust, mechanically stable materials. Silica and poly(N-isopropylacrylamide) (PNIPAAm) precursors were copolymerized in the presence of surfactant supramolecular assemblies to form hybrid gels with ordered nanostructure. This method was less complicated and results in enhanced reversible transport properties compared to previous approaches noted herein. In this study, the thermoresponsive polymer, PNIPAAm, was incorporated into polymerizing silica networks using the coupling agent 3-methacryloxypropyltrimethoxysilane. The hydration transition of PNIPAAm associated with its lower critical solution temperature (LCST) in aqueous solution was retained in the hydrated silica matrices and was used to control the permeability of membranes and molecular release behavior of particles. This report presents new methods for formation of hybrid silica/PNIPAAm membranes and particles, characterization of these materials, and documentation of reversible molecular transport properties of these new hybrid materials.

This study was conducted to determine the effect of Se supplementation and source on the Se status of horses. Eighteen 18-mo-old nonexercised horses were randomly assigned within sex to 1 of 3 treatments: 1) control (CTRL, no supplemental Se, 0.15 mg of Se/kg of total diet DM); 2) inorganic Se (INORG, CTRL + 0.45 mg of Se/kg of total diet DM from NaSeO3); or organic Se [ORG, CTRL + 0.45 mg of Se/kg of total diet DM from zinc-L-selenomethionine (Availa Se, Zinpro, Corp., Eden Prairie, MN)]. Horses were acclimated to the CTRL diet (7.1 kg of DM alfalfa hay and 1.2 kg of DM concentrate per horse daily) for 28 d. After the acclimation period, the appropriate treatment was top-dressed on the individually fed concentrate for 56 d. Jugular venous blood samples were collected on d 0, 28, and 56. Middle gluteal muscle biopsies were collected on d 0 and 56. Muscle and plasma were analyzed for Se concentrations. Glutathione peroxidase activity was measured in muscle (M GPx-1), plasma (P GPx-3), and red blood cells (RBC GPx-1). Data were analyzed as a repeated measures design. Mean plasma Se concentration on d 28 and 56 was greater (P < 0.05) for Se-supplemented horses compared with CTRL horses, and tended (P < 0.1) to be greater in ORG vs. INORG on d 28. Mean muscle Se concentration and P GPx-3 activities increased (P < 0.05) from d 0 to 56 but were not affected by treatment. Mean RBC GPx-1 activity tended to be greater (P < 0.1) in ORG than INORG or CTRL horses on d 28, and tended to be greater (P < 0.1) for INORG compared with ORG horses on d 56. Mean RBC GPx-1 activity of INORG and ORG horses was not different from that of CTRL on d 56. Mean M GPx-1 activity decreased (P < 0.01) from d 0 to 56. In conclusion, zinc-L-selenomethionine was more effective than NaSeO3 at increasing plasma Se concentration from d 0 to 28; however, both supplemental Se sources had a similar effect by d 56. No difference in Se status due to Se supplementation or source could be detected over a 56-d supplementation period by monitoring middle gluteal muscle Se, M GPx-1, or P GPx-3. Results for RBC GPx-1 also were inconclusive relative to the effect of Se supplementation and source.

Encephalitozoon intestinalis is a microsporidian species that infects the intestinal mucosal epithelium, primarily in immunodeficient individuals. The present study employed undifferentiated and differentiated human colonic carcinoma cell lines to determine if this parasite species infected polarized epithelial cells by spore phagocytosis or by impalement with the deployed spore polar tube. Apical surface spore attachment differed between cell lines such that SW480>HT-29>Caco-2>HCT-8, with attachment being greater to undifferentiated Caco-2 cells than differentiated cells and greater to partially differentiated HCT-8 cells than differentiated HCT-8 cells. Attachment was inhibited by chondroitin sulfate A, suggesting that it was mediated by host cell sulfated glycoaminoglycans. Infection rates 3 days postinfection paralleled spore attachment in the various cell lines. The undifferentiated cell line SW480 and undifferentiated Caco-2 and HCT-8 cells exhibited modest spore phagocytosis while the more differentiated cell line HT29 and differentiated Caco-2 and HCT-8 cells did not. All cell lines were impaled by the polar tubes of germinating spores. When normalized to the number of spores attached to the apical membrane, such impalement was greatest in the more differentiated Caco-2 and HCT-8 cells. The host cell apical surface influenced parasite spore germination, as in populations of large undifferentiated Caco-2 cells to which >3 spores had attached, the frequency distribution of the percentages of spores germinated per cell was bimodal, indicating that the surface of some cells favored germination, while others did not. This study suggests that phagocytosis is not a biologically significant mode of infection in differentiated intestinal epithelial cells.

Highly dispersed gold particles (<2 nm) were synthesized within the pores of mesoporous silica with pore sizes ranging from 2.2 to 6.5 nm and different pore structures (2D-hexagonal, 3D-hexagonal, and cubic). The catalysts were reduced in flowing H2 at 200 degrees C and then used for CO oxidation at temperatures ranging from 25 to 400 degrees C. The objective of this study was to investigate the role of pore size and structure in controlling the thermal sintering of Au nanoparticles. Our study shows that sintering of Au particles is dependent on pore size, pore wall thickness (strength of pores), and pore connectivity. A combination of high-resolution TEM/STEM and SEM was used to measure the particle size distribution and to determine whether the Au particles were located within the pores or had migrated to the external silica surface.

Maxiban and Monteban are 2 products marketed by Elanco Animal Health. They contain narasin and are used for the prevention of coccidiosis in chickens. Products used in the European market must be regularly re-registered with new data to support label claims. This study was undertaken as part of such a re-registration effort. A method for the determination of narasin in poultry tissue was previously registered with the authorities; however, a method with more environmentally friendly solvents was desired. The Canadian Food Inspection Agency accomplished this goal and published an improved method. In order to register the method with European authorities as the official Elanco method for determination of narasin, Elanco scientists were required to provide validation data for all edible poultry tissues. This paper shows the validation of the method to detect residues of narasin using solid-phase extraction followed by liquid chromatographic analysis utilizing post-column derivatization.

Pores arranged in a two-dimensional hexagonal structure inside spherical mesoporous silica particles help to prevent the thermal sintering of gold nanoparticles compared to straight pores in MCM-41.

Comparative biochemical and histopathological evidence suggests that a deficiency in the glycogen branching enzyme, encoded by the GBE1 gene, is responsible for a recently identified recessive fatal fetal and neonatal glycogen storage disease (GSD) in American Quarter Horses termed GSD IV. We have now derived the complete GBE1 cDNA sequences for control horses and affected foals, and identified a C to A substitution at base 102 that results in a tyrosine (Y) to stop (X) mutation in codon 34 of exon 1. All 11 affected foals were homozygous for the X34 allele, their 11 available dams and sires were heterozygous, and all 16 control horses were homozygous for the Y34 allele. The previous findings of poorly branched glycogen, abnormal polysaccharide accumulation, lack of measurable GBE1 enzyme activity and immunodetectable GBE1 protein, coupled with the present observation of abundant GBE1 mRNA in affected foals, are all consistent with the nonsense mutation in the 699 amino acid GBE1 protein. The affected foal pedigrees have a common ancestor and contain prolific stallions that are likely carriers of the recessive X34 allele. Defining the molecular basis of equine GSD IV will allow for accurate DNA testing and the ability to prevent occurrence of this devastating disease affecting American Quarter Horses and related breeds.

1. Since high concentrations of zinc are associated with reduced crop yields, environmental concerns are emerging regarding zinc accumulation in areas where poultry production is prevalent. This study investigates growth performance and zinc utilisation during the life cycle of broilers when diets were supplemented with various concentrations of zinc from two different sources. 2. A total of 740 Cobb 500 1-d-old male broiler chicks was randomly distributed into 88 battery cages. Excreta were collected over a 48-h period on d 10, 17, 24, 31, 38, and 45 to measure zinc excretion on a dry matter basis. 3. During the 45-d experimental period, broilers were given a maize-soybean meal basal diet (30 mg/kg zinc) supplemented with 0, 20, 40, and 80 mg/kg of zinc from zinc sulphate, Availa Zn amino acid complex, or an equal combination of zinc sulphate and Availa Zn amino acid complex. 4. Progressive additions of zinc to the basal diet significantly increased body weight gain of broilers, but did not affect feed conversion or mortality. Optimum body weight gain was achieved at 80 mg/kg supplemental zinc, which exceeds the NRC recommendations of 40 mg/kg. 5. During each excreta collection period, increased supplemental zinc concentrations significantly increased zinc excretion. However, the zinc source did not influence zinc utilisation. 6. As compared with zinc sulphate supplementation, the cumulative zinc excretion data indicate that adding zinc to the basal diet in the form of Availa Zn amino acid complex decreased zinc excretion.

This research was conducted to evaluate immunity (experiments 1 to 3), cardiac function, and ascities resistance (experiment 4) of progeny chicks from broiler breeders fed diets differing in trace metal level and source. Broiler breeders received a control diet (75 mg of Zn and 83 mg of Mn added/kg of diet), the control diet supplemented with inorganic Zn (75 mg/kg of diet) and Mn (80 mg/kg of diet), the control diet supplemented with organic Zn (75 mg/kg of diet) and inorganic Mn (80 mg/kg of diet), or the control diet supplemented with organic Zn (75 mg/kg of diet) and Mn (80 mg/kg of diet) in experiments 1, 2, and 3. In experiment 4, the control diet and diet supplemented with organic sources of Zn and Mn were fed to broiler breeders. Immune organ weights, circulating granulocytes vs. agranulocytes, CD4 and CD8 positive T cells, cutaneous basophil hypersensitivity, and antibody titers to SRBC and breeder vaccinations were measured in progeny. Some supplemental mineral treatments increased (P < or = 0.05) cutaneous basophil hypersensitivity and relative bursa weight. All supplemental mineral treatments increased (P < or = 0.05) relative thymus weight. In experiment 4, electrocardiograph, pulse oximetry, heart rate, hematocrits, ventricle weights, and ascites incidence were measured in progeny reared in a cold-stress environment. The supplemental organic minerals increased (P < or = 0.05) left ventricle plus septum and total ventricular weights. Although progeny ascites incidence did not differ between breeder mineral treatments, breeders fed supplemental Zn and Mn sired progeny with improved cardiac functional capacity and some improvements in immunity.

1. Environmental accumulation of zinc and copper is becoming a concern in areas having intensive animal production. This study examined performance and excretion of broiler chicks given diets supplemented with graded concentrations of zinc and copper from three different sources. 2. Two experiments were conducted, each utilising 570 1-d-old chicks. In Experiment 1, chicks were given diets containing supplemental zinc concentrations from 40 to 120 mg/kg supplied as zinc sulphate, a zinc amino acid complex (Availa Zn), or a combination of zinc sulphate and Availa Zn with each contributing one-half of the total supplemental zinc. In Experiment 2, broiler chicks were given diets with graded concentrations of supplemental copper ranging from 4 to 12 mg/kg from copper sulphate, a copper amino acid complex (Availa Cu), or a combination of copper sulphate and Availa Cu with each contributing one-half of the total supplemental copper. 3. During the 17-d experimental period, mineral concentration or source did not influence body weight, feed conversion, or the incidence of mortality from 1 to 17 d. 4. Decreasing dietary zinc concentration from 120 to 40 mg/kg reduced zinc excretion by 50%. 5. Copper excretion was decreased by 35% as supplemental copper was reduced from 12 to 4 mg/kg. 6. Reducing dietary zinc and copper concentrations can potentially decrease the accumulation of heavy metals in the environment without compromising bird performance.

Comparative biochemical and histopathological data suggest that a deficiency in the glycogen branching enzyme (GBE) is responsible for a fatal neonatal disease in Quarter Horse foals that closely resembles human glycogen storage disease type IV (GSD IV). Identification of DNA markers closely linked to the equine GBE1 gene would assist us in determining whether a mutation in this gene leads to the GSD IV-like condition. FISH using BAC clones as probes assigned the equine GBE1 gene to a marker deficient region of ECA26q12-->q13. Four other genes, ROBO2, ROBO1, POU1F1, and HTR1F, that flank GBE1 within a 10-Mb segment of HSA3p12-->p11, were tightly linked to equine GBE1 when analyzed on the Texas A&M University 5000 rad equine radiation hybrid panel, while the GLB1, MITF, RYBP, and PROS1 genes that flank this 10-Mb interval were not linked with markers in the GBE1 group. A polymorphic microsatellite (GBEms1) in a GBE1 BAC clone was then identified and genetically mapped to ECA26 on the Animal Health Trust full-sibling equine reference family. All Quarter Horse foals affected with GSD IV were homozygous for an allele of GBEms1, as well as an allele of the most closely linked microsatellite marker, while a control horse population showed significant allelic variation with these markers. This data provides strong molecular genetic support for the candidacy of the GBE1 locus in equine GSD IV., (Copyright 2003 S. Karger AG, Basel)

Seven related Quarter Horse foals that died by 7 weeks of age were examined for glycogen branching enzyme (GBE) deficiency. Clinical signs varied from stillbirth, transient flexural limb deformities, seizures, and respiratory or cardiac failure to persistent recumbency. Leukopenia (5 of 5 foals) as well as high serum creatine kinase (CK; 5 of 5), aspartate transaminase (AST; 4 of 4), and gamma glutamyl transferase (GGT; 5 of 5) activities were present in most foals, and intermittent hypoglycemia was present in 2 foals. Gross postmortem lesions were minor, except for pulmonary edema in 2 foals. Muscle, heart, or liver samples from the foals contained abnormal periodic acid Schiff's (PAS)-positive globular or crystalline intracellular inclusions in amounts proportional to the foal's age at death. Accumulation of an unbranched polysaccharide in tissues was suggested by a shift in the iodine absorption spectra of polysaccharide isolated from the liver and muscle of affected foals. Skeletal muscle total polysaccharide concentrations were reduced by 30%, but liver and cardiac muscle glycogen concentrations were normal. Several glycolytic enzyme activities were normal, whereas GBE activity was virtually absent in cardiac and skeletal muscle, as well as in liver and peripheral blood cells of affected foals. GBE activities in peripheral blood cells of dams of affected foals and several of their half-siblings or full siblings were approximately 50% of controls. GBE protein in liver determined by Western blot was markedly reduced to absent in affected foals, and in a half-sibling of an affected foal, it was approximately one-half the amount of normal controls. Pedigree analysis also supported an autosomal recessive mode of inheritance. The affected foals have at least 2,600 half-siblings. Consequently, GBE deficiency may be a common cause of neonatal mortality in Quarter Horses that is obscured by the variety of clinical signs that resemble other equine neonatal diseases.

Objective: To determine whether an alteration in calcium regulation by skeletal muscle sarcoplasmic reticulum, similar to known defects that cause malignant hyperthermia (MH), could be identified in membrane vesicles isolated from the muscles of Thoroughbreds with recurrent exertional rhabdomyolysis (RER)., Sample Population: Muscle biopsy specimens from 6 Thoroughbreds with RER and 6 healthy (control) horses., Procedures: RER was diagnosed on the basis of a history of > 3 episodes of exertional rhabdomyolysis confirmed by increases in serum creatine kinase (CK) activity. Skeletal muscle membrane vesicles, prepared by differential centrifugation of muscle tissue homogenates obtained from the horses, were characterized for sarcoplasmic reticulum (SR) activities, including the Ca2+ release rate for the ryanodine receptor-Ca2+ release channel, [3H]ryanodine binding activities, and rate of SR Ca2+-ATPase activity and its activation by Ca2+., Results: Time course of SR Ca2+-induced Ca2+ release and [3H]ryanodine binding to the ryanodine receptor after incubation with varying concentrations of ryanodine, caffeine, and ionized calcium did not differ between muscle membranes obtained from control and RER horses. Furthermore, the maximal rate of SR Ca2+-ATPase activity and its affinity for Ca2+ did not differ between muscle membranes from control horses and horses with RER., Conclusions and Clinical Relevance: Despite clinical and physiologic similarities between RER and MH, we concluded that RER in Thoroughbreds does not resemble the SR ryanodine receptor defect responsible for MH and may represent a novel defect in muscle excitation-contraction coupling, calcium regulation, or contractility.

Thirty-two Suffolk wether lambs were fed for 84 d in a 2 x 2 factorial experiment using two levels of dietary protein (9.0 to 12.1% CP, low protein, LP; or 12.8 to 14.4% CP, high protein, HP) and supplemental Cr (none, C; or 400 ppb Cr as chromium tripicolinate, Cr). At 14- to 21-d intervals, lambs were weighed, and jugular blood samples were collected. Mean ADG and carcass weight (P > .10) did not differ. In lambs fed HP, Cr reduced liver weight and increased kidney weight (P < .01). Lambs fed HP had elevated plasma urea N (PUN; P < .01) and albumin (P < .04). During an i.v. epinephrine challenge on d 43, plasma cortisol declined in lambs fed Cr (Cr x time, P < .03) and in lambs fed LP (CP x time, P < .001). An i.v. glucose tolerance test conducted 3 h later showed that supplemental Cr decreased glucose clearance rate in lambs fed HP (CP x Cr, P < .10) but not in lambs fed LP. On d 62, PUN was increased in lambs fed HP (P < .001) between 0 and 3 h postprandial, and there was a Cr x CP interaction (P < .04). Postprandial plasma NEFA declined with Cr vs C (Cr x time, P < .07) and with HP vs LP (CP x time, P < .10). By d 66, lambs fed Cr had an elevated (P < .03) blood platelet and fibrinogen content. Chromium increased erythrocyte count in lambs fed HP (Cr x CP, P < .08), and isolated peripheral lymphocytes had greater blastogenic response to 4 microg/mL of phytohemagglutinin (Cr x CP, P < .001). The lymphocyte response to pokeweed mitogen (.2 microg/mL) was reduced in lambs fed Cr (P < .10). In the present experiment, Cr supplementation had minimal and inconsistent effects on production and metabolic criteria of lambs.