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The rearing environment for pullets should mirror their later production environment as closely as possible. However, existing perch space recommendations are based on data for laying hens rather than pullets. This study explores the impact of perch space allowances on the welfare of Canadian laying strain pullets raised to 18 wk. Two trials were conducted with 1,032 Lohmann Brown-Lite (LB) and LSL-Lite (LW) pullets each. A randomized complete block (trial) design was used with a 4 (perch space) × 2 (strain) factorial arrangement. Birds were raised in 16 floor pens (3 × 3 m each; 897.67 cm 2 per bird). Wooden perches were provided from d 1, allowing 6, 9, 12, or 15 cm perch space per pullet. Data were tested for normality, and log+1 transformed if necessary. Significance was declared at P≤0.05. Data were collected for basic health and functioning (body weight, mortality, pullet width, keel bone damage, and tibia bone parameters), affective states (comb damage, heterophil-to-lymphocyte (H/L) ratios, and behavior), and natural living (perch usage and jumping success). Perch space did not affect mortality, keel bone damage, tibia breaking strength, comb damage, or H/L ratios. At 16 and 18 wk, LB body weight slightly increased while the LW body weight decreased with increasing perch space (P=0.05 and 0.02). At 3 wk, pullets spent a higher percentage of time wing flapping on the perch when provided 15 cm compared to 6 cm (P=0.04). During the scotoperiod at 18 wk, a higher percentage of birds perched with 12 and 15 cm perch space than 6 cm (P<0.01). Pullet width at 18 wk was 12.52 cm (sitting) and 11.66 cm (standing) for the LB and 13.85 cm (sitting) and 12.94 cm (standing) for the LW. Overall, perch space allowance had minor effects on the measured welfare indicators. Based on pullet width, a minimum of 12.5 cm for LW and 13.9 cm for LB pullets should be provided to allow all pullets to sit on the perch simultaneously., Competing Interests: Disclosures The authors declare no conflict of interest., (Copyright © 2024. Published by Elsevier Inc.)

The effects of light intensity (L) are not well studied in pullets. Our research objective was to study the effect of L on navigational success, behavior, and welfare of two pullet strains (S). In two repeated trials, a 3 × 2 × 4 factorial arrangement tested three L (10, 30, 50 lux) and two S (Lohmann Brown-Lite (LB), LSL-Lite (LW)) at four ages. One thousand eight hundred pullets/S (0-16 wk) were randomly assigned to floor pens within light-tight rooms (three pens/S/room, four rooms/L) containing four parallel perches and a ramp. Data collection included jumping frequency and success (24h continuous sampling), novel object tests (fear), heterophil to lymphocyte (H/L) ratios (stress), and behavior (instantaneous scan sampling) during photoperiods. L did not affect injurious behavior, fear, or H/L. Pullets reared at 50 lux spent more time preening than at 10 lux. Pullets reared at 10 lux spent more time wall pecking than at 50 lux. Time spent standing and preening and total number and accuracy of jumping increased with age. Pullets reared at 30 lux had higher jumping frequency than at 10 lux; accuracy was not affected. LW jumped more than LB, but with similar success. LB spent more time exploring and scored higher in the fear and stress assessments, suggesting S differences.

The objective of this research was to evaluate activation of the ileal brake in broiler chickens using diets containing semi-purified wheat (WS; rapidly and highly digested) and pea (PS; slowly and poorly digested) starch. Diets were formulated to contain six WS:PS ratios (100:0, 80:20, 60:40, 40:60, 20:80, 0:100) and each starch ratio was fed to 236 Ross 308 male broilers housed in 4 litter floor pens. At 28 d of age, the effect of PS concentration was assessed on starch digestion, digestive tract morphology, and digesta pH and short-chain fatty acid (SCFA) concentration. Glucagon-like peptide-1 (GLP-1) and peptide tyrosine-tyrosine (PYY) status were assessed in serum (ELISA) and via gene expression in jejunal and ileal tissue (proglucagon for GLP-1). Data were analyzed using regression analyses, and significance was accepted at P ≤ 0.05. Increasing dietary PS resulted in reduced starch digestibility in the small intestine, but had no effect in the colon. Crop content pH responded quadratically to PS level with an estimated minimum at 55% PS. Total SCFA increased linearly in the crop with PS level, but changed in a quadratic fashion in the ileum (estimated maximum at 62% PS). Ceacal SCFA concentrations were highest for the 80 and 100% PS levels. The relative empty weight (crop, small intestine, colon), length (small intestine) and content (crop jejunum, Ileum) of digestive tract sections increased linearly with increasing PS concentration. Dietary treatment did not affect serum GLP-1 or PYY or small intestine transcript abundance. In conclusion, feeding PS increased the presence of L-cell activators (starch, SCFA) and increased trophic development and content of the digestive tract, suggestive of L-cell activation. However, no direct evidence of ileal brake activation was found by measuring venous blood levels of GLP-1 or PYY or corresponding gene expression in small intestine tissue., Competing Interests: We have the following competing interests: This study was funded in part by Aviagen North America, Sofina Foods Inc., Prairie Pride Natural Foods Ltd., Chicken Farmers of Saskatchewan, Canadian Poultry Research Council, Poultry Industry Council (Canada), Saskatchewan Broiler Hatching Egg Producer's Marketing Board, Saskatchewan Egg Producers and Saskatchewan Turkey Producers' Marketing Board. There are no patents, products in development or marketed products to declare. This does not alter our adherence to all the PLOS ONE policies on sharing data and materials.

Myo-inositol (inositol) affects memory, and the incidence of depression and anxiety in mammals. An experiment was designed to determine if pure inositol (0.16%), or high levels of phytase (3000 FTU/kg) affect the behaviour of fully beaked Lohmann LSL lite hens fed amino acid sufficient (19% crude protein (CP)) and deficient diets (16% CP), from 19 to 59 weeks of age. The data collected included live-scan behaviour observations and novel object (NO) tests (both at 1, 10 and 40 weeks of the trial); heterophil-to-lymphocyte (H/L) ratios (week 1 and week 40 of the trial); end of trial feather cover, and comb and skin lesions; and daily mortality. Reducing CP increased sitting by 2.5%. Inositol, but not phytase, reduced the latency to peck at the NO by 300 sec. Inositol reduced vent feather cover by 12% and tended to increase mortality by 13%. No effects on H/L ratio, and comb or skin lesions were found. In conclusion, regardless of the source, inositol reduced vent feather cover, while it tended to increase mortality. Only pure inositol reduced fearfulness in laying hens.

Infrared beak treatment has less of a negative impact on laying hen welfare compared to other methods of beak treatment; however, it is still not fully understood how infrared beak treatment affects the beak tissue during the first few days post treatment. The objective of this study was to examine the histology of infrared beak treated vs. untreated beaks of 2 strains of layer chicks during early life. One-hundred Lohmann Brown-Lite (LB) and 100 Lohmann LSL-Lite (LW) chicks were obtained; 50 chicks per strain were infrared beak treated post hatch (IR) with the remainder being sham untreated controls (C). Data collected included presence of beak sloughing, length, and histology. Histology slides were analyzed and scored on a scale of 0 to 4, with 0 indicating no lesions and 4 indicating severe inflammation. Sloughing of the treated beak tissue began at 10 days and was complete by 20 days. IR pullets had shorter beak lengths once sloughing was initiated and less overall beak growth. No differences in healing scores were found between treated LB and LW beaks; all treated LB beaks were healed by 21 days while some LW beaks still showed inflammation. Overall, infrared beak treatment was effective at reducing beak growth post treatment. Healing occurred post treatment in both strains as evident by complete regeneration of the epithelium and a reduction in inflammation.

Dietary starch with lower rate and extent of digestion improves broiler feed efficiency, but previous results might have been confounded by non-starch components of the grains. Therefore, the objective of this research was to study the effects of starch digestion on broilers using semi-purified starch. Semi-purified wheat (WS, rapidly digested) and pea (PS, slowly digested) starch were combined to create 6 WS:PS ratios (100:0, 80:20, 60:40, 40:60, 20:80, and 0:100) in starter, grower and finisher diets. Each treatment was fed to Ross 308 male (2,124) and female (2,376) broilers housed in 72 L floor pens from 0 to 31 d of age to measure performance and meat yield relative to live weight. On day 33, the effects of diet on 8 h feed withdrawal was assessed in 20 males per treatment. Data were analyzed with ANOVA and linear and quadratic regression analyses using SAS 9.4. Significance was accepted at P ≤ 0.050. Body weight gain declined linearly with increasing PS. Male feed intake decreased with increasing PS, but PS did not affect female feed intake. Mortality corrected gain:feed ratio was quadratically influenced by diet (estimated maximum at 25% PS). Breast meat increased linearly with PS, while fat pad and breast and thigh skin decreased linearly. Quadratic responses were found for thigh meat and whole drum (estimated maximum values at 56 and 54% PS, respectively). Males grew faster, ate more, and had higher mortality than females. They also had heavier pectoralis major, thigh bone, and whole drum, while females had heavier pectoralis minor and more breast and thigh skin. After feed withdrawal, digesta content decreased linearly with time in all sections, except for the crop and duodenum, which declined quadratically. Ileal digesta pH increased linearly with time, while crop and caecal pH decreased for 2 h before steadily increasing. Diet did not affect digestive tract emptying or digesta pH. In conclusion, dietary PS maximized feed efficiency at 25% PS and linearly improved breast meat yield, but did not affect digesta clearance after feed withdrawal., (© 2019 Poultry Science Association Inc.)

The inclusion of starch with a lower rate and extent of digestion has proven to be beneficial in broiler production. However, less is known about its effect on laying hen performance. Therefore, 6 diets were formulated to produce differing ratios (0:100, 20:80, 40:60, 60:40, 80:20, and 100:0) of semi-purified wheat starch (rapidly digested) and pea starch (PS; slowly digested). Each diet was fed to 120 conventionally caged Lohmann LSL lite hens in groups of 12, from 27 to 46 wk of age, and its effects on performance and feather covering were assessed. Data were analyzed by regression analysis, and the significance level was chosen at P ≤ 0.05. Hen-day egg production (HDP) was high (97.05%) and unaffected by PS concentration for 0 to 10 wk of the trial, followed by a quadratic-shaped HDP, with an estimated maximum at 67% PS concentration, for the 10 to 20 wk period of the trial. Overall (0 to 20 wk), HDP increased linearly with PS. No effect on egg weight was found (average weight = 59.6±2.1 g), but eggshells were thickest at the 55% PS concentration. Body weight gain was affected by PS concentration and maximized at 49% PS. Body weight uniformity and mortality were not affected by dietary treatment. Feed intake increased linearly with PS from 102 to 109 g/hen/d, while feed: egg mass ratio was minimal at 26% PS. Using a scale from 1 to 4 per body part (neck, wings, back, vent and breast), only neck feather cover increased linearly with PS, from 3.0 (0% PS) to 3.2/4 (100% PS). However, back and total feather coverage showed a trend (P = 0.054 and P = 0.079 respectively) to increase linearly with PS as well (from 3.7 to 3.9/4 and 15.6 to 16.6/20, respectively). In summary, feeding PS at low to intermediate concentrations improved laying hen performance. Further research should focus on the mechanisms involved in this effect., (© 2019 Poultry Science Association Inc.)

Sepsis represents a significant cause of mortality in intensive care units. Early diagnosis of sepsis is essential to increase the survival rate of patients. Among others, C-reactive protein (CRP) is commonly used as a sepsis marker. In this work we introduce immune precipitation combined with microchip capillary gel electrophoresis (IP-MCGE) for the detection and quantification of CRP in serum samples. First high-abundance proteins (HSA, IgG) are removed from serum samples using affinity spin cartridges, and then the remaining proteins are labeled with a fluorescence dye and incubated with an anti-CRP antibody, and the antigen/antibody complex is precipitated with protein G-coated magnetic beads. After precipitation the complex is eluted from the beads and loaded onto the MCGE system. CRP could be reliably detected and quantified, with a detection limit of 25 ng/μl in serum samples and 126 pg/μl in matrix-free samples. The overall sensitivity (LOQ = 75 ng/μl, R(2) = 0.9668) of the method is lower than that of some specially developed methods (e.g., immune radiometric assay) but is comparable to those of clinically accepted ELISA methods. The straightforward sample preparation (not prone to mistakes), reduced sample and reagent volumes (including the antibodies), and high throughput (10 samples/3 h) are advantages and therefore IP-MCGE bears potential for point-of-care diagnosis., (Copyright © 2015 Elsevier Inc. All rights reserved.)

Purpose: Urothelial cells, including bladder cancer (BCa) cells, represent a highly valuable but challenging target for localized antineoplastic therapy. This study describes a novel, biomimetic approach to improve intravesical drug delivery, based on glycan-specific targeting. In direct analogy to the invasion mechanism used by uropathogenic bacteria, we evaluate the potential of lectin bioconjugates to facilitate binding and uptake of large payload molecules at this penetration-hostile barrier., Methods: Wheat germ agglutinin (WGA) served as a targeting ligand and was covalently coupled to fluorescein-labeled bovine serum albumin (fBSA), yielding multivalent protein bioconjugates. Cytoadhesion, uptake and intracellular processing were characterized on a panel of urothelial cell lines of non-malignant and malignant origin., Results: Conjugation to WGA rendered the fBSA payload protein strongly cytoadhesive, with a clear preference in binding to cancerous cells. The highly specific, lectin-mediated recognition process was followed by rapid internalization, and extensive but non-exclusive accumulation in acid and LAMP-2-positive compartments. Stage of malignancy and mechano-structural cell configuration were important determinants for the sorting between different processing pathways., Conclusion: Lectin-bioconjugates allow for triggering endogenous uptake routes and influencing the intracellular distribution in BCa cells. They hold considerable promise for enhancing the delivery of small molecule drugs and complex biomolecules in intravesical therapy.

A sensitive and selective analytical method for the determination and quantification of endogenous β-galactosidase in crude E. coli cell lysates by immunoprecipitation combined with automated microchip capillary gel electrophoresis (IP-MCGE) with laser-induced fluorescence (LIF) detection was developed. Total cell lysates were derivatized minimally with a fluorescence dye, incubated with anti-β-galactosidase antibodies, and the antigen/antibody complex was precipitated with protein G-coated magnetic beads. After capturing the complex, it was eluted from the beads under denaturing conditions and loaded directly onto a multisample microchip for analysis. The effects of antibody selection and the importance of preclearing steps were studied in detail. For quantification, an external calibration through spiking pure β-galactosidase into E. coli lysate was performed. Recovery rates of immunoprecipitation after spiking experiments and the amount of unknown endogenous β-galactosidase in E. coli lysates were determined. As proof of principle, E. coli cultures grown on nutrition media with several glucose/lactose ratios were analyzed. Differences in the expression level of β-galactosidase could be detected and measured with the developed method. Detected amounts of β-galactosidase in different culture media correlated with the β-galactosidase activities in these cultures., (Copyright © 2011 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

A novel pre-chip fluorescent derivatization method is presented for protein sizing and quantification by microchip CGE. The derivatization reaction employed a water-soluble and stable fluorescent dye and was performed under conditions that favored the formation of homogeneous reaction products. The method delivered in terms of protein sizing similar results as microchip CGE with on-chip staining but showed an extended linear dynamic range for protein quantification encompassing four orders of magnitude. The sensitivity of the method was similar to standard silver-stained planar gels. The characterization of derivatization reaction products by MS and preparative isoelectric focusing indicated that a constant degree of dye molecule tagging was obtained over a broad range of protein/dye ratios. The method allowed detecting and quantifying an impurity spiked into an antibody preparation down to a level of 0.05%. Advantages of this method compared with CGE approaches with pre-column derivatization include a shorter analysis time and an increased robustness and ease of use.