Genetic background and production periods shape the microRNA profiles of the gut in laying hens

There is growing evidence of the importance of miRNAs for intestinal functional properties and nutritional uptake. Comparative miRNAs profiles of the jejunal mucosa were established against two genetic backgrounds (Lohmann Brown-Classic (LB) and Lohmann LSL-Classic (LSL), which are similar in egg production but differ in physiological traits including mineral utilization, along the production periods of laying hens. The target genes of miRNAs higher expressed in LB vs. LSL (miR-126-3p, miR-214, miR-24-3p, miR-726-5p, miR-29b-3p) were enriched for energy pathways at all stages. The target genes of the miRNAs higher in LSL (miR-1788-5p, miR-103-3p, miR-22-5p, miR-221-3p, miR-375) were more enriched for immune and the bone signalling pathways. The most prominent expression differences were between 16 and 24 weeks of age before and after onset of laying. Our results evidence central roles of intestinal miRNAs as regulators of gene expression, influencing intestinal homeostasis and adaptation to environment in different strains and production phases.     

Differential Gene Expression from Genome-Wide Microarray Analyses Distinguishes Lohmann Selected Leghorn and Lohmann Brown Layers

The Lohmann Selected Leghorn (LSL) and Lohmann Brown (LB) layer lines have been selected for high egg production since more than 50 years and belong to the worldwide leading commercial layer lines. The objectives of the present study were to characterize the molecular processes that are different among these two layer lines using whole genome RNA expression profiles. The hens were kept in the newly developed small group housing system Eurovent German with two different group sizes. Differential expression was observed for 6,276 microarray probes (FDR adjusted P-value <0.05) among the two layer lines LSL and LB. A 2-fold or greater change in gene expression was identified on 151 probe sets. In LSL, 72 of the 151 probe sets were up- and 79 of them were down-regulated. Gene ontology (GO) enrichment analysis accounting for biological processes evinced 18 GO-terms for the 72 probe sets with higher expression in LSL, especially those taking part in immune system processes and membrane organization. A total of 32 enriched GO-terms were determined among the 79 down-regulated probe sets of LSL. Particularly, these terms included phosphorus metabolic processes and signaling pathways. In conclusion, the phenotypic differences among the two layer lines LSL and LB are clearly reflected in their gene expression profiles of the cerebrum. These novel findings provide clues for genes involved in economically important line characteristics of commercial laying hens.     

Effects of ambient temperature and methionine supplementation of a low protein diet on the performance of laying hens

The effects of ambient temperature and the supplementation of methionine to a low protein diet on egg production, egg quality, blood constituents and nitrogen excretion of laying hens were studied. The objective was to derive an environmental friendly feed formulation for warm climate. Seventy-two 29-week-old commercial White Leghorn hens of Babcock ISA white strain were used in this trial. The design is a completely randomized design with a 2×3 factors arrangement of treatments. Two constant ambient temperatures were 24±1°C and 34±1°C with 85% relative humidity. The three dietary treatments were 170 g kg⁻¹ crude protein, 140 g kg⁻¹ crude protein and 140 g kg⁻¹ crude protein supplemented with methionine 1.4 g kg⁻¹. Hens were allotted into six groups according to egg production and body weight. Birds were raised in individual wire cages for the experimental feeding period of five weeks. At the end of the feeding trial, one replicate of laying hens (four birds) from each treatment were selected for a four-day metabolic study for the daily collection of the excreta. The blood samples were withdrawn from the wing vein for analysis of hematocrit, blood glucose, sodium, potassium, magnesium, calcium, phosphorus and uric acid. Experimental results indicate that increases in ambient temperature significantly depress feed intake, egg production, egg weight and live weight of laying hens. High ambient temperature also caused inferior egg quality, including shell weight, shell thickness, shell breaking strength and specific gravity. Ambient temperature also changes the egg components with heavier egg albumin and yolk in the low-temperature group. Increasing ambient temperature also caused an increase in pH value in the plasma. This increase revealed a trend of depressed glucose (P<0.05) in the plasma of the laying hens. The dietary treatments, however, did not significantly influence feed intakes. Except egg weight, laying hens that were fed with the low protein (140 g kg⁻¹) with methionine supplemented diet produced similar numbers of egg and feed conversion as the layers fed with the high protein diet. The low protein with methionine supplemented diet produced significantly lighter eggs than the high dietary protein diet under the high ambient temperature, but produced heavier egg under the low ambient temperature. The concentration of uric acid in the plasma and nitrogen in the excreta of the high protein group was significantly higher than the other two low protein dietary groups (P<0.05).     

Effects of feeding deoxynivalenol (DON)-contaminated wheat to laying hens and roosters of different genetic background on the reproductive performance and health of the newly hatched chicks

A total of 216 23-week-old laying hens from two different genetic backgrounds (half of the birds were Lohmann brown [LB] and [LSL] hens, respectively) and 24 adult roosters were assigned to a feeding trial to study the effect of increasing concentrations of deoxynivalenol (DON) in the diet (0, 5, 10 mg/kg) on the reproductive performance of hens and roosters, and the health of the newly hatched chicks. Hatchability was adversely affected by the presence of DON in LB hens’ diet, while the hatchability of the LSL chicks was significantly higher than LB chicks. An interaction effect between DON in the hens’ diet and the breed was noticed on fertility, as the fertility was decreased in the eggs of LB hens receiving 10 mg/kg DON in their diet and increased in the eggs of LSL hens fed 10 mg/kg DON. Moreover, spleen relative weight was significantly decreased in the chicks hatched from eggs of hens fed contaminated diets, while gizzard relative weight was significantly decreased in LB chicks with 10 mg/kg DON in their diet compared with the control group. On the other hand, the chicks’ haematology and organ histopathology were not affected by the dietary treatment. Additionally, the presence of DON in the roosters’ diet had no effect on fertility (the percentage of fertile eggs of all laid eggs). Consequently, the current results indicate a negative impact of DON in LB hens’ diet on fertility and hatchability, indicating that the breed of the hens seems to be an additional factor influencing the effect of DON on reproductive performance of the laying hens.     

Effect of inorganic phosphate supplementation on egg production in Hy-Line Brown layers fed 2000 FTU/kg phytase

Phytase has long been used to decrease the inorganic phosphorus (Pi) input in poultry diet. The current study was conducted to investigate the effects of Pi supplementation on laying performance, egg quality and phosphate–calcium metabolism in Hy-Line Brown laying hens fed phytase. Layers (n = 504, 29 weeks old) were randomly assigned to seven treatments with six replicates of 12 birds. The corn–soybean meal-based diet contained 0.12% non-phytate phosphorus (nPP), 3.8% calcium, 2415 IU/kg vitamin D₃ and 2000 FTU/kg phytase. Inorganic phosphorus (in the form of mono-dicalcium phosphate) was added into the basal diet to construct seven experimental diets; the final dietary nPP levels were 0.12%, 0.17%, 0.22%, 0.27%, 0.32%, 0.37% and 0.42%. The feeding trial lasted 12 weeks (hens from 29 to 40 weeks of age). Laying performance (housed laying rate, egg weight, egg mass, daily feed intake and feed conversion ratio) was weekly calculated. Egg quality (egg shape index, shell strength, shell thickness, albumen height, yolk colour and Haugh units), serum parameters (calcium, phosphorus, parathyroid hormone, calcitonin and 1,25-dihydroxyvitamin D), tibia quality (breaking strength, and calcium, phosphorus and ash contents), intestinal gene expression (type IIb sodium-dependent phosphate cotransporter, NaPi-IIb) and phosphorus excretion were determined at the end of the trial. No differences were observed on laying performance, egg quality, serum parameters and tibia quality. Hens fed 0.17% nPP had increased (P < 0.01) duodenum NaPi-IIb expression compared to all other treatments. Phosphorus excretion linearly increased with an increase in dietary nPP (phosphorus excretion = 1.7916 × nPP + 0.2157; R² = 0.9609, P = 0.001). In conclusion, corn–soybean meal-based diets containing 0.12% nPP, 3.8% calcium, 2415 IU/kg vitamin D₃ and 2000 FTU/kg phytase would meet the requirements for egg production in Hy-Line Brown laying hens (29 to 40 weeks of age).     

Do laying hens eat and forage in excreta from other hens?

Worldwide, farm animals are kept on litter or foraging substrate that becomes increasingly soiled throughout the production cycle. For animals like laying hens, this means that it is likely they would scratch, forage and consume portions of excreta found in the litter or foraging substrate. However, no study has investigated the relative preference of laying hens for foraging and consumption of feed mixed with different percentages of excreta. A total of 48 White Leghorn laying hens of two strains, a commercial strain (Lohmann LSL-Lite (LSL), n=24) and UCD-003 strain (susceptible to liver damage, n=24), were individually housed and given access to feed mixed with increasing percentages of hen excreta (0%, 33%, 66% and 100% excreta diets) and corn as a luxury food reward (four corn kernels per diet daily). The amount of substrate and number of corn kernels consumed from each diet was recorded for a period of 3 weeks. Both LSL and UCD-003 hens preferred to consume and forage in diets with 0% excreta, followed by 33% and finally diets containing 66% and 100% excreta. Despite the presence of excreta-free diets, birds consumed on average 61.3 g per day of the diets containing excreta. Neither physical health, measured by plasma enzyme activity levels, nor cognitive differences, assessed by recalling a visual discrimination task, was associated with relative feeding or foraging preference. In conclusion, this study demonstrated a clear preference for feeding and foraging on substrate without excreta in laying hens. However, considering the amount of excreta diets consumed, further studies are needed to understand the causes and consequences of excreta consumption on physiological and psychological functioning, and how this information can be used to allow adjustments in the management of foraging substrates in farmed birds.     

Effects of enzyme-treated soy protein on performance,digestive enzyme activity and mRNA expression of nutrient transporters of laying hens fed different nutrient density diets

It has been shown that enzyme-treated plant protein can increase performance and promote intestinal health, and save dietary protein. However, our understanding of the effects of enzyme-treated soy protein on performance and intestine function in laying hens, and its rational use, remains limited. The experiment was conducted to study the effect of enzyme-treated soy protein (ETSP) in different nutrient density diets on performance, egg quality, digestive enzyme activity and mRNA expression of amino acid transporters of laying hens. A total of 1 200 Lohmann laying hens (52 wk of age) was randomly divided into a 3 × 2 factorial design that included three nutrient levels: [positive control (PC), metabolisable energy (ME): 2 680 kcal/kg, CP: 15.5%; negative control 1 (NC1), ME: 2 630 kcal/kg, CP: 15%; negative control 2 (NC2), ME:2 580 kcal/kg, CP: 14.5%] and 2 ETSP levels (0 and 0.5%) for 12 weeks. Each treatment had 10 replicates with 20 birds. With the decrease of dietary nutrition density, egg production rate (P = 0.07) and feed conversion ratio (FCR) (P = 0.06) were reduced. Yolk colour was decreased, and yolk index was increased. Supplemented ETSP improved FCR (P = 0.05) and qualified egg rate (P < 0.05). The mass loss rate of egg was decreased after storage for 30 days (P < 0.05). An interaction between nutrient density and ETSP was observed on albumen height and Haugh unit (P < 0.05), and the effects were most noticeable in hens fed 0.5% ETSP in NC2 group. An increase in the activity of trypsin in duodenum (P < 0.05) and the relative expressions of jejunum peptide transporter 1 (PepT1) (P < 0.05) and B⁰ system neutral amino acid transport carrier (B⁰AT) mRNA (P < 0.01) was observed during ETSP supplementation. The nutrient density and ETSP supplementation had no significant effect on microbiota in the cecal contents. Overall, the results in this study indicated that the ME decreased 100 kcal/kg and CP decreased 1% in diet of laying hens had a decreasing trend on production performance, no effects on enzyme activity, amino acid transporter mRNA, and gut microbiota, whereas 0.5% ETSP can increase activity of trypsin, PepT1 and B⁰AT mRNA relative expressions, and improve FCR, qualified egg rate.     

Effects of phytoestrogen supplementation in the feed on the shell gland of laying hens at the end of the laying period

Shell quality decreases as laying hens age and the aim of present study was to investigate how a supplement of daidzein, a natural phytoestrogen in soya, affects key factors in the shell gland and eggshell quality in late-stage laying hens. Hybrids of Lohmann Selected Leghorn (LSL) and Lohmann Brown (LB), received either a daidzein diet (50mg/kg feed) or a control diet from 60 to 72 weeks of age. Both the total number of capillaries and capillaries with carbonic anhydrase (CA) activity were higher in the LSL hybrid than in the LB. After daidzein supplementation the number of CA positive capillaries was unaffected in the LSL but increased in the LB hybrid indicating a higher sensitivity to daidzein in this hybrid. Estrogen receptor alpha and beta (ERα, ERβ) were localized and the complete picture of the two ERs can now be described in shell gland of domestic hens. Nuclear and cytoplasmic staining was generally stronger for ERβ, while membrane associated staining was present only for ERα. Interestingly, capillary endothelium contained only ERβ and since estrogen regulation of CA is well documented, the presence of an endothelial ER provides one possible route for the increase in CA positive capillaries found in LB hybrids. Eggshell quality or egg production was not affected by daidzein supplementation. The hybrids used in this study showed anatomical differences and reacted differently to daidzein supplementation, but if this can be explained by the divergences in ERβ localization noted between the hybrids remains to be clarified.     

Effects of dietary Bacillus subtilis and inulin supplementation on performance,eggshell quality,intestinal morphology and microflora composition of laying hens in the late phase of production

Eighty Lohmann White laying hens were used to investigate the effect of dietary inclusion of Bacillus subtilis and inulin, individually or in combination, on egg production, eggshell quality, tibia traits, Ca retention, and small intestine morphology and microflora composition from 64 to 75 weeks of age. Hens were randomly distributed into 4 treatment groups, with 5 replicates per treatment and 4 hens per replicate. Treatment groups were fed basal diet (control), basal diet plus 1 g/kg B. subtilis (2.3 × 10⁸ cfu/g), basal diet plus 1 g/kg inulin, or basal diet plus a synbiotic combination of 1 g/kg B. subtilis (2.3 × 10⁸ cfu/g) and 1 g/kg inulin. Dietary supplementation of B. subtilis, inulin or synbiotic improved (P<0.05) feed conversion, egg performance, eggshell quality and calcium retention compared with the control. B. subtilis and synbiotic groups exhibited the highest (P<0.05) increase in egg production and egg weight. Inulin and synbiotic groups exhibited the highest (P<0.05) increase in eggshell thickness and eggshell calcium content, and the lowest (P<0.05) eggshell deformations. Unmarketable eggs were 8.4% (P<0.05) of the total eggs produced by the control group compared to 3.5%, 1.7%, and 1.5% for the B. subtilis, inulin and synbiotic groups, respectively. Tibia density, ash, and Ca content increased (P<0.05) by inulin and synbiotic inclusions, compared with the control. B. subtilis, inulin, and their synbiotic combination increased (P<0.05) villus height and crypt depth in all intestinal segments, compared with the control. B. subtilis and inulin modulated the ileal and caecal microflora composition by decreasing (P<0.05) numbers of Clostridium and Coliforms and increasing (P<0.05) numbers of bifidobacteria and lactobacilli, compared with the control. Colonization of the beneficial microflora along with increasing the villi–crypts absorptive area were directly associated with the improvements in performance and eggshell quality. It can be concluded that egg production and eggshell quality of laying hens can be improved (P<0.05) in the late phase of production by dietary inclusion of B. subtilis and inulin.     

Development of locomotion over inclined surfaces in laying hens

The purpose of the present study was to evaluate locomotor strategies during development in domestic chickens (Gallus gallus domesticus); we were motivated, in part, by current efforts to improve the design of housing systems for laying hens which aim to reduce injury and over-exertion. Using four strains of laying hens (Lohmann Brown, Lohmann LSL lite, Dekalb White and Hyline Brown) throughout this longitudinal study, we investigated their locomotor style and climbing capacity in relation to the degree (0 to 70°) of incline, age (2 to 36 weeks) and the surface substrate (sandpaper or wire grid). Chicks and adult fowl performed only walking behavior to climb inclines ⩽40° and performed a combination of wing-assisted incline running (WAIR) or aerial ascent on steeper inclines. Fewer birds used their wings to aid their hind limbs when climbing 50° inclines on wire grid surface compared with sandpaper. The steepness of angle achieved during WAIR and the tendency to fly instead of using WAIR increased with increasing age and experience. White-feathered strains performed more wing-associated locomotor behavior compared with brown-feathered strains. A subset of birds was never able to climb incline angles >40° even when using WAIR. Therefore, we suggest that inclines of up to 40° should be provided for hens in three-dimensional housing systems, which are easily negotiated (without wing use) by chicks and adult fowl.     

New pentafluorophenyl complexes with phosphine-amide ligands

A series of nickel(II) and palladium(II) complexes containing one or two pentafluorophenyl ligands and the phosphino-amides o-Ph₂PC₆H₄CONHR [R = ⁱPr (a), Ph (b)] displaying different coordination modes have been synthesised. The chelating ability of these ligands and the influence of both coligands and the metal centre in their potential hemilabile behaviour have been explored. The crystal structure of (b) has been determined and reveals N–H⋯O intermolecular hydrogen bonding. Bis-pentafluorophenyl derivatives [M(C₆F₅)₂(o-Ph₂PC₆H₄CO-NHR)] [M = Ni; R = ⁱPr (1a); R = Ph (1b); M = Pd; R = ⁱPr (2a); R = Ph (2b)] in which (a) and (b) act as rigid P, O-chelating ligands were readily prepared from the labile precursors cis-[M(C₆F₅)₂(PhCN)₂]. X-ray structures of (1a), (1b) and (2a) have been established, allowing an interesting comparative structural discussion. Dinuclear [{Pd(C₆F₅)(tht)(μ-Cl)}₂] reacted with (a) and (b) yielding the monopentafluorophenyl complexes [Pd(C₆F₅)Cl{PPh₂(C₆H₄–CONH–R)}] (R = ⁱPr (3a), Ph (3b)) that showed a P, O-chelating behaviour of the ligands, confirmed by the crystal structure determination of (3a). New cationic palladium(II) complexes in which (a) and (b) behave as P-monodentate ligands have been synthesised by reacting them with [{Pd(C₆F₅)(tht)(μ-Cl)}₂], stoichiometric Ag(O₃SCF₃) and external chelating reagents such as cod [Pd(C₆F₅)(cod){PPh₂(C₆H₄-CONH-R)}](O₃SCF₃)(R = ⁱPr (4a), Ph (4b)) and 2,2′-bipy [Pd(C₆F₅)(bipy){PPh₂(C₆H₄-CONH-R)}](O₃SCF₃) (R = ⁱPr (5a), Ph (5b)). When chloride abstraction in [{Pd(C₆F₅)(tht)(μ-Cl)}₂] is promoted by means of a dithioanionic salt as dimethyl dithiophospate in the presence of (a) or (b), the corresponding neutral complexes [Pd(C₆F₅){S(S)P(OMe)₂}{PPh₂(C₆H₄-CONH-R)}] (R = ⁱPr (6a), Ph (6b)) were obtained.     

Genome-wide association studies for mottled eggs in chickens using a high-density single-nucleotide polymorphism array

Mottled eggs in layer chickens are gaining increasing attention because of the economic impact on the egg industry caused by the reduced sale value of commodity eggs. However, the genetic architecture underlying mottled eggs is not well understood. The genetic architecture underlying the mottled egg trait was investigated using genome-wide association studies (GWAS) by high-density arrays, using a total of 407 pink eggs and 799 blue eggs from an F₂ resource population generated by crossing Dongxiang Blue-shelled and White Leghorn chickens. The mottled egg score in blue eggs was found to be higher than that in pink eggs. The single-nucleotide polymorphism heritability of mottled egg at laying day and storage for 7 days was 0.18 and 0.20, respectively. Bivariate GWAS provided 29 significant loci, mainly located on GGA2, GGA3, GGA8, GGA10, GGA15, GGA17, and GGA23, affecting mottled egg on laying day. Candidate genes RIMS2, SLC25A32, RIMBP2, VPS13B, and RGS3 were obtained for mottled eggshell by bivariate GWAS and gene annotation. Our findings provide new insights into the genetic architecture of mottled egg in hens, and demonstrate that a genomic selection method would be profitable for breeding out the mottled egg trait.     

Effects of phytase supplementation on production performance,egg and bone quality,plasma biochemistry and mineral excretion of layers fed varying levels of phosphorus

Excessive fecal excretion of phosphorus (P) has increasingly become an environmental issue due to oversupply of P in layer rations, and thus it is imperative to minimize safety margins for P to ensure the sustainability of the egg industry. In this study, a 12-week feeding trial (22 to 34 weeks of age) was conducted to evaluate the effects of phytase supplementation on production performance, plasma biochemistry, egg and bone quality and P excretion of laying hens fed various levels of non-phytate P (NPP). Forty-eight Lohmann white laying hens were randomly allocated to one of six corn–soybean meal–oat-based diets: diets containing 2.0, 2.5 or 3.0 g/kg NPP without phytase, and diets containing 1.0, 1.5 or 2.0 g/kg NPP with phytase (1 000 U/kg diet) where phytase inclusion was expected to provide 1.0 g/kg NPP to laying hens, thus making the phytase-unsupplemented treatment served as a control for the phytase-supplemented treatment accordingly. Productive performance was recorded during the experimental period. Blood and egg samples were collected, and digestibility studies were conducted at weeks 6 and 12 of the experiment. Bone mineralization was evaluated at the end of the experiment. Egg weight and egg production, feed consumption, BW and feed conversion ratio of laying hens fed lower NPP diets supplemented with phytase were comparable to those of hens fed high NPP phytase-unsupplemented controls. Eggshell thickness, specific gravity, Haugh units, tibia bone mineral density, tibia ash percent, plasma P and other biochemical parameters were not significantly different among dietary treatments. Total P intake, excretion and retention were affected by diet (P < 0.001), but its deposition in eggs was not significantly different. Contrast analysis further showed that total P excretion of phytase present vs phytase absent was averagely reduced by 40.4 mg/hen per day (P < 0.01). Moreover, total P excretion was linearly (P < 0.01) reduced with lowering dietary NPP, and this relationship was similar regardless of whether phytase was supplemented or not. The results from this study indicated that NPP levels in laying hen diets could be reduced to 1.0 g/kg (excluding the portion of NPP released by phytase) with the inclusion of phytase, without negative effects on production performance and health of the hens, thereby diminishing P excretion into environment.     

Accuracy of laying hens in jumping upwards and downwards between perches in different light environments

Eighty Lohmann Brown layer hens were reared in floor pens fitted with perches from 1 day of age. After the peak of lay (25 weeks of age), their ability to jump to and from perches in different light environments was assessed in two experiments using digital video techniques that allowed detailed measurements of take-off, flight and landing behaviours.In Experiment 1, birds jumped up or down (angles of 10 and 18°) between two horizontal perches that were separated by a gap of 60 cm under different lighting conditions (5, 10 or 20 lux; incandescent or high- or low-frequency fluorescent). In Experiment 2, the horizontal distance was increased to 80 cm and contrast between perch and background was varied. Fifty-two hens (65%) achieved the training criterion for Experiment 1: jumping a 60 cm gap five times of which three in a row. Thirty-two of these hens (62%) subsequently failed to achieve the 80 cm jump criterion for Experiment 2.In Experiment 1, birds took off sooner (P<0.001), turned their heads further downward relative to the landing perch (P<0.001), beat their wings more often during flight (P<0.001), and took longer to achieve balance (P<0.001) when jumping downward rather than upward. In Experiment 2, refusals to jump, and clumsy or missed landings, occurred more frequently on downward than upward jumps (P<0.001). These behaviours were unaffected by lighting conditions, or by the amount of contrast between the perch and its background.In conclusion, behaviours indicating inaccurate control of landing were more frequent on downward than on upward jumps (P<0.001). However, these behaviours were not affected by light intensity, light type or contrast. The results suggest that there is a difference between the two types of jumps in the risk of injury to hens housed in aviaries where perches must be used to gain access to resources.     

Glycopeptide dendrimer colchicine conjugates targeting cancer cells

Screening of a 65,536-member one-bead-one-compound (OBOC) combinatorial library of glycopeptide dendrimers of structure ((βGal)_n + 1X⁸X⁷X⁶X⁵)₂DapX⁴X³X²X¹(β-Gal)_m (βGal = β-galactosyl-thiopropionic acid, X^8–1 = variable amino acids, Dap = l-2,3-diaminopropionic acid, n, m = 0, or 1 if X⁸ = Lys resp. X¹ = Lys) for binding of Jurkat cells to the library beads in cell culture, resynthesis and testing lead to the identification of dendrimer J1 (βGal-Gly-Arg-His-Ala)₂Dap-Thr-Arg-His-Asp-CysNH₂ and related analogues as delivery vehicles. Cell targeting is evidenced by FACS with fluorescein conjugates such as J1F. The colchicine conjugate J1C is cytotoxic with LD₅₀ = 1.5 μM. The β-galactoside groups are necessary for activity, as evidenced by the absence of cell-binding and cytotoxicity in the non-galactosylated, acetylated analogue AcJ1F and AcJ1C, respectively. The pentagalactosylated dendrimer J4 βGal₄(Lys-Arg-His-Leu)₂Dap-Thr-Tyr-His-Lys(βGal)-Cys) selectively labels Jurkat cell as the fluorescein derivative J4F, but its colchicine conjugate J4C lacks cytotoxicity. Tubulin binding assays show that the colchicine dendrimer conjugates do not bind to tubulin, implying intracellular degradation of the dendrimers releasing the active drug.

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The association between plumage damage and feather-eating in free-range laying hens

Severe feather-pecking (SFP) persists as a highly prevalent and detrimental behavioural problem in laying hens (Gallus gallus domesticus) worldwide. The present experiment investigated the association between feather-eating and plumage damage, a consequence of SFP, in groups of free-range, ISA Brown laying hens. Single feathers were placed on the floor of the home pens. Feathers were sourced from seven different birds. A total of 50 birds in six pens with extensive plumage damage were compared with birds in six control pens with little plumage damage at 41 to 43 weeks of age (n=12 pens, 600 hens). Birds in pens with extensive plumage damage ingested more feathers (F=8.1, DF=1, 8, P=0.02), and also showed shorter latencies to peck at (χ²=54.5, DF=1, P<0.001), and ingest feathers (χ²=55.6, DF=1, P<0.001). Birds ingested feathers from a bird in the free-range facility, in which the testing took place, more quickly than from a bird housed in a separate cage facility (χ² = 39.0, DF=6, P<0.001). A second experiment investigated the predictive relationship between feather-eating and plumage damage. Feathers were presented to 16 pens of 50 pullets prior to the development of plumage damage, at 15 weeks of age, and then to the same hens after plumage damage had become prominent, at 40 weeks of age. Birds had a higher probability of ingesting feathers (F=142.0, DF=1, 231, P<0.001), pecked feathers more times (F=11.24, DF=1, 239, P<0.001), and also pecked (χ² = 127.3, DF=1, P<0.001) and ingested (χ²=189.3, DF=1, P<0.001) the feathers more quickly at 40 than 15 weeks of age. There was a trend for an interaction, where birds pecked feathers from the rump more times than feathers from the back at 40 weeks of age (F=3.46, DF=1, 237, P=0.06). However, a lack of variability in plumage damage between pens in this experiment precluded investigation of the predictive relationship. The results from the present study confirm the association between feather-eating and plumage damage, and suggest that birds may prefer feathers from particular body areas and from particular hens. Future experiments should focus on elucidating whether feather-eating may act as a predictor of SFP.     

Dietary inclusion of raw faba bean instead of soybean meal and enzyme supplementation in laying hens: Effect on performance and egg quality

An experiment was conducted with 160 Hisex Brown laying hens to evaluate the effect of different inclusion levels of faba bean (FB) and enzyme supplementation on productive performance and egg quality parameters. The experimental diets consisted of five levels of FB: 0% (control), 25%, 50%, 75% and 100%, substituting soybean meal (SBM), and two levels of enzyme supplementation (0 or 250 mg/kg). Each dietary treatment was assigned to four replicate groups and the experiment lasted 22 weeks. A positive relationship (P < 0.05) was found between FB inclusion and body weight (BW) change of hens when compared to those of the control treatment. Enzyme supplementation significantly affected the final hens’ BW. Feed consumption (FC) of hens was statistically increased with increasing FB level up to 50%. Supplementing dietary enzyme mixture at 250 mg/kg led to improvement in FC at all studied ages (P < 0.05). Inclusion of 25% or 50% FB in diets had no adverse effects on feed conversion ratio (FCR) compared to the higher FB inclusion levels (75% or 100%). Egg weight (EW), egg number (EN) and egg mass (EM) were significantly (P < 0.05) influenced by FB inclusion in diet during the entire experimental periods, except for EN and EM at 20–24 weeks of age. Egg productive parameters were not influenced by enzyme mixture supplementation (P > 0.05). The main effect of FB levels replacing for SBM affected (P < 0.05) yolk and shell percentages, yolk index, yolk to albumen ratio, shell thickness and egg shape index. It can be concluded that FB and enzyme supplementation could be included in hens diet at less than 50% instead of SBM to support egg productive performance, however higher raw FB levels negatively affected egg production indices and quality.     

Hormone Substitution after Gastric Bypass Surgery in Patients with Hypopituitarism Secondary to Craniopharyngioma

Objective: Craniopharyngiomas (CPs) are benign brain tumors presenting frequently in childhood and are treated by surgery with or without radiotherapy. About 50% of cured patients suffer from eating disorders and obesity due to hypothalamic damage, as well as hypopituitarism, necessitating subsequent hormone substitution therapy. Gastric bypass surgery has been reported to be an efficient treatment strategy for morbid hypothalamic obesity. However, so far it is unknown whether oral hormone substitution is affected by impaired intestinal drug absorption, potentially leading to severe hypopituitarism or pituitary crisis.Methods: Four morbidly obese CP patients with panhypopituitarism treated by gastric bypass surgery were included in this retrospective analysis. Dosages of hormone substitution therapy, blood concentrations of hormones, potential complications of impaired drug absorption, and anthropometric characteristics were investigated pre- and postoperatively after 6 to 14 months and 13 to 65 months.Results: In all CP patients (3 female/1 male; baseline body mass index, 49 ± 7 kg/m²), gastric bypass resulted in distinct weight loss (-35 ± 27 kg). In follow-up examinations, mean daily dosage of thyroid hormone (levothyroxine_baseline 156 ± 44 μg/day versus levothyroxine_follow-up 150 ± 30 μg/day), hydrocortisone (hydrocortisone_baseline 29 ± 12 mg/day versus hydrocortisone_follow-up 26 ± 2 mg/day), growth-hormone (somatotropin_baseline 0.9 ± 0.5 mg/day versus somatotropin_follow-up 1.0 ± 0.4 mg/day), and desmopressin (desmopressin_baseline 222 ± 96 μg/day versus desmopressin_follow-up 222 ± 96 μg/day) substitution was unchanged. No patient developed adrenal insufficiency. Oral thyroid/hydrocortisone absorption testing performed in 1 patient indicated sufficient gastrointestinal drug absorption after bariatric surgery.Conclusion: Our preliminary results suggest that oral hormone substitution therapy is not impaired following gastric bypass operation in CP patients with morbid obesity, indicating that it might be a safe and effective treatment strategy.Abbreviations:BMI = body mass indexCP = craniopharyngiomafT4 = free thyroxineGH = growth hormoneIGF-1 = insulin-like growth factor 1     

The impacts of fermented feed on laying performance,egg quality,immune function,intestinal morphology and microbiota of laying hens in the late laying cycle

Fermented feed has the potential to improve poultry gastrointestinal microecological environment, health condition and production performance. Thus, the present study was undertaken to explore the effects of fermented feed on the laying performance, egg quality, immune function, intestinal morphology and microbiota of laying hens in the late laying cycle. A total of 360 healthy Hy-Line Brown laying hens aged 80 weeks were used to conduct a 56-day study. All hens were randomly separated into two treatment groups, with five replicates of 36 hens each as follows: basal diet containing 0.0% fermented feed (CON) and 20% fermented feed (FF). Subsequent analyses revealed that fermented feed supplementation was associated with significant increases in laying rates together with reduced broken egg rates and feed conversion ratio for hens in FF group (P < 0.05). There were additionally significant increases in both albumen height and Haugh unit values in hens following fermented feed supplementation (P < 0.05). Fermented feed was also associated with increases in duodenal, jejunal and ileac villus height (P < 0.05). Laying hens fed fermented feed had higher immune globulin (Ig)A, IgG, IgM levels (P < 0.01,) and higher interleukin 2, interleukin 6, tumour necrosis factor α and interferon γ (P < 0.05) concentrations than CON. Analysis of the microbiota in these laying hens revealed the alpha diversity was not significantly affected by fermented feed supplementation. Firmicutes abundance was reduced in caecal samples from FF hens relative to those from CON hens (30.61 vs 35.12%, P < 0.05). At the genus level, fermented feed was associated with improvements in relative Lactobacillus, Megasphaera and Peptococcus abundance and decreased Campylobacter abundance in laying hens. These results suggest that fermented feed supplementation may be beneficial to the laying performance, egg quality, immunological function, intestinal villus growth and caecal microecological environment of laying hens at the end of the laying cycle.     

Radiosynthesis and first evaluation in mice of [18F]NS14490 for molecular imaging of α7 nicotinic acetylcholine receptors

[¹⁸F]NS14490, a new potential radiotracer for neuroimaging of α7 nicotinic acetylcholine receptors (α7 nAChRs), was synthesized and evaluated in vitro and in vivo. Radioligand binding studies using [³H]methyllycaconitine and NS14490 as competitor showed a good target affinity (K_i,α7 = 2.5 nM) and a high selectivity towards other nAChRs. Radiosynthesis of [¹⁸F]NS14490 was performed by two different labelling procedures: a two-step synthesis using a prosthetic group, which led to 7% labelling yield, and the convenient direct nucleophilic substitution of the corresponding tosylate precursor, which resulted in 70% labelling yield. After optimisation of the isolation, purification and formulation process, biodistribution studies were performed in CD-1 mice. The brain uptake of [¹⁸F]NS14490 was comparably low (0.16% ID g^?1 wet weight at 5 min p.i.). The radiotracer showed a high metabolic stability in plasma and brain. Also, the target specificity was proven by pre-administration of a highly affine α7 ligand providing a rationale basis for further in vivo evaluation.