This study assess how well diclofenac (DCF) can be separated from aqueous solution using potassium permanganate-modified eggshell biosorbent (MEB). The MEB produced was characterised using XRD, FTIR, and SEM. Batch experiments were conducted to examine and assess the impact of contact time, adsorbent dosage, initial concentration, and temperature on the adsorption capacity of the MEB in the DCF sequestration. The best parameters to obtained 95.64% DCF removal from liquid environment were 0.05 g MEB weight, 50 mg/L initial concentration, and 60 min contact time at room temperature. The maximum DCF sequestration capacity was found to be 159.57 mg/g with 0.05 g of MEB at 298 K. The adsorption isotherm data were more accurately predicted by the Freundlich model, indicating a process of heterogeneous multilayer adsorption. The results of the kinetic study indicated that the pseudo-second-order kinetic models best matched the experimental data. The findings revealed that the dynamic of DCF entrapment is largely chemisorption and diffusion controlled. Based on the values of thermodynamic parameters, the process is both spontaneous and endothermic. The primary processes of DCF sorption mechanism onto the MEB were chemical surface complexation, hydrogen bonding, π-π stacking, and electrostatic interactions. The produced MEB showed effective DCF separation from the aqueous solution and continued to have maximal adsorption capability even after five regeneration cycles. These findings suggest that MEB could be highly efficient adsorbent for the removal of DCF from pharmaceutical wastewater.

\"On-farm hatching\" is one of the proposed alternatives to conventional hatchery-hatching. This solution reduces distress and improves the welfare of the chicks around the hatching period. Therefore, it seemed interesting to compare conventional hatchery and \"on-farm\" hatching in terms of microbiological and microclimatic conditions. Hatching eggs (Ross 308) were incubated in a commercial hatchery. The control group (HH, 683 eggs) hatched in a conventional hatcher, while the other eggs were transported into the experimental chicken-hall for on-farm hatching, and set in pens directly on litter (OL, 667 eggs) or plastic trays (OT, 678 eggs). One-day-old chicks were also placed in the experimental hall. Microclimatic parameters were controlled every 12 h. The microbiological status of the surface of the eggshells and the litter was assessed based on the total number of aerobic mesophilic microorganisms and also the selected individual genus/species of bacteria. The hatchability of HH was 96.4% in comparison to 93.9% and 95.8% for OL and OT, respectively (P > 0.05). On the other hand, 2.1% of the HH chicks were found injured/dead, while only 0.2-0.3% of the on-farm groups were. The total number of aerobic mesophilic microflora on the surface of as-hatched shells was 4.93 ± 0.629 log CFU/g in HH, while only 1.14 ± 0.995 and 1.93 ± 1.709 log CFU/g in OL and OT, respectively (P < 0.001). Similarly, the total count of bacteria in the litter in the on-farm hatched pens was 1.9-fold lower than in pens set with HH chicks (P < 0.001). In summary, on-farm hatching results in hatchability that is no worse than in a conventional hatcher, while the microbiological status of as-hatched eggshells and litter is significantly better. Therefore, on-farm hatching seems to provide appropriate environmental conditions for newly hatched chicks and poses no epizootic risk.

UNASSIGNED: The eggshell and the eggshell membrane (ESM) are significant by-products of the poultry industry and are being utilized for various valuable purposes in health care, like soft tissue healing and pain alleviation. The aim and objective of our study are to assess the effect of the eggshell membrane on alveolar bone regeneration after tooth extraction. A total of 40 extraction sockets (bilateral) among 20 patients were assessed clinically for healing, and radiographic parameters of bone density and socket volume were assessed on CBCT at baseline, 3 months, and 6 months. Advanced platelet-rich fibrin was created from 5 ml of autologous blood from the patient and centrifuged for 15 minutes at 1500 RPM/168 RCF. The commercially available powdered form of egg shell membrane was used in the study. Based on the randomized allotment (coin-flip), A-PRF alone or A-PRF mixed with eggshell membrane was placed inside the extraction socket and was stabilized using 3-0 silk sutures. It was ob-served that wound healing was uneventful in all 20 patients. No evidence of dry sockets or allergic reactions was noted in any patient. Statistical analysis was done using the un-paired t-test and Mann-Whitney U test with SPSS version 20.0. P<0.05 was considered significant. On comparison of the mean bone density at baseline, 3 months, and 6 months, the socket density in the eggshell with the PRF group was higher compared to the control group. To conclude, eggshell membrane has good regenerative properties and excellent osteogenic capacity; therefore, it could be a useful graft due to its low cost, abundant availability, and simple application.

The eggshell is a composite and highly ordered structure formed by biomineralization. Besides other functions, it has a vital and intricate role in the protection of an embryo from various potentially harsh environmental conditions. Solid-state nuclear magnetic resonance (SSNMR) has been used for detailed structural investigations of the chicken, tinamou, and flamingo eggshell materials. 31P NMR spectra reveal that hydroxyapatite and β-tricalcium phosphate in the ratio 3:2 represent major constituents of phosphate species in the eggshells. All three eggshells exhibit similar spectra, except for the line widths, which implies different structural order of phosphate species in the chicken, tinamou, and flamingo eggshells. 1H NMR spectra for these materials are comparable, differentiating overlapped peaks in three spectral regions at around 7, 4-5, and 1-2 ppm. These spectral regions have been attributed to protons from NH or CaHCO3, water, and possibly isolated monomeric water molecules or hydroxyl groups in calcium-deficient hydroxyapatite. 1H-13C CP MAS NMR revealed the presence of organic matter in the form of lipids and proteins. Two overlapped resonances in the carbonyl region at around 173 and 169 ppm are assigned to the carbonyls of the peptide bonds and the bicarbonate unit in calcite, respectively. Fourier-transform infrared spectroscopy (FTIR) spectra confirmed the presence of structural units detected in the NMR spectra.

Osteoarthritis (OA) is a chronic degenerative joint disease that causes chronic pain, swelling, stiffness, disability, and significantly reduces the quality of life. Typically, OA is treated using painkillers and non-steroidal anti-inflammatory drugs (NSAIDs). While current pharmacologic treatments are common, their potential side effects have prompted exploration into functional dietary supplements. Recently, eggshell membrane (ESM) has emerged as a potential functional ingredient for joint and connective tissue disorders due to its clinical efficacy in relieving joint pain and stiffness. Despite promising clinical evidence, the effects of ESM on OA progression and its mechanism of action remain poorly understood. This study evaluated the efficacy of Ovomet®, a powdered natural ESM, against joint pain and disease progression in a monosodium iodoacetate (MIA)-induced rodent model of OA in mice and rats. The results demonstrate that ESM significantly alleviates joint pain and attenuates articular cartilage destruction in both mice and rats that received oral supplementation for 5 days prior to OA induction and for 28 days thereafter. Interestingly, ESM significantly inhibited mRNA expression levels of pro-inflammatory cytokines including tumor necrosis factor alpha (TNF-α), interleukin-1β (IL-1β), and interleukin-6 (IL-6), as well as inflammatory mediators, cyclooxygenase-2 (COX-2), and inducible nitric oxide synthase in the knee joint cartilage at the early stage of OA, within 7 days after OA induction. However, this effect was not observed in the late stage at 28 days after OA induction. ESM further attenuates the induction of protein expression for cartilage-degrading enzymes like matrix metalloproteinase (MMPs) 3 and 13, and a disintegrin and metalloproteinase with thrombospondin motifs 5 (ADAMTS-5), in the late-stage. In addition, MIA-induced reduction of the protein expression levels of cartilage components, cartilage oligomeric matrix protein (COMP), aggrecan (ACAN) and collagen type II α-1 chain (COL2α1), and cartilage extracellular matrix (ECM) synthesis promoting transcriptional factor SRY-Box 9 (SOX-9) were increased via ESM treatment in the cartilage tissue. Our findings suggest that Ovomet®, a natural ESM powder, is a promising dietary functional ingredient that can alleviate pain, inflammatory response, and cartilage degradation associated with the progression of OA.

Eggshell color plays important biological roles and attracts the attention of both egg retailers and researchers. However, whether non-coding RNAs are involved in pigment deposition among different eggshell colors remains unknown. In this study, RNA sequencing was used to analyse the uterine gland transcriptome (CircRNA and miRNA) of Changshun chicken blue-shell hens producing four different eggshell color eggs including dark blue PK(DB) and light blue (LB), dark brown and greenish (between blue and pink, DP) and pink (p). We found that miR-192-x, targeting SLC16a7, was expressed in DB, DP, and LB groups compared with the PK group, which indicates that miR-192-x may play a role in the blue eggshell color. KEGG and GO analyses showed that the \"metabolic pathways\" with targeted genes such BLVRA and HMOX1 were detected in dark and light blue color eggshell chickens, which confirms the different ratios of biliverdin and HO-1 involved in the deposition of blue color. As annotated by connectivity analysis, RASGRF1 and RASGRF2, belonging to the RASGRF family, are involved in the Ras signaling pathway, which plays an important role in cell growth, differentiation, metastasis and apoptosis. Our findings enrich the database of circRNA, miRNAs and genes for chicken uterine tissue, which will be useful in accelerating molecular selection for blue eggshell color layers.

Green eggs are mainly caused by inserting an avian endogenous retrovirus (EVA-HP) fragment into the SLCO1B3 gene. Although the genotypes for this insertion allele are consistent, eggshell color (ESC) may vary after a peak laying period; light-colored eggs are undesired by consumers and farmers and result in financial loss, so it is necessary to resolve this problem. miRNAs are small non-coding RNAs that exert essential functions in animal development and diseases. However, the regulatory miRNAs and detailed molecular mechanisms regulating eggshell greenness remain unclear. In the present study, we determined the genotype of green-eggshell hens through the detection of a homozygous allele insertion in the SLCO1B3 gene. The shell gland epithelium was obtained from green-eggshell hens that produced white and green shell eggs to perform transcriptome sequencing and investigate the important regulatory mechanisms that influence the ESC. Approximately 921 miRNAs were expressed in these two groups, which included 587 known miRNAs and 334 novel miRNAs, among which 44 were differentially expressed. There were 22 miRNAs that were significantly upregulated in the green and white groups, respectively, which targeted hundreds of genes, including KIT, HMOX2, and several solute carrier family genes. A Gene Ontology enrichment analysis of the target genes showed that the differentially expressed miRNA-targeted genes mainly belonged to the functional categories of homophilic cell adhesion, gland development, the Wnt signaling pathway, and epithelial tube morphogenesis. A KEGG enrichment analysis showed that the Hedgehog signaling pathway was significantly transformed in this study. The current study provides an overview of the miRNA expression profiles and the interaction between the miRNAs and their target genes. It provides valuable insights into the molecular mechanisms underlying green eggshell pigmentation, screening more effective hens to produce stable green eggs and obtaining higher economic benefits.

Three-dimensional (3D) printing, an additive manufacturing technique, is increasingly used in the field of tissue engineering. The ability to create complex structures with high precision makes the 3D printing of this material a preferred method for constructing personalized and functional materials. However, the challenge lies in developing affordable and accessible materials with the desired physiochemical and biological properties. In this study, we used eggshell microparticles (ESPs), an example of bioceramic and unconventional biomaterials, to reinforce thermoplastic poly(ε-caprolactone) (PCL) scaffolds via extrusion-based 3D printing. The goal was to conceive a sustainable, affordable, and unique personalized medicine approach. The scaffolds were fabricated with varying concentrations of eggshells, ranging from 0 to 50% (w/w) in the PCL scaffolds. To assess the physicochemical properties, we employed scanning electron microscopy, Fourier-transform infrared spectroscopy, thermogravimetric analysis, differential scanning calorimetry, and X-ray diffraction analysis. Mechanical properties were evaluated through compression testing, and degradation kinetics were studied through accelerated degradation with the remaining mass ranging between 89.4 and 28.3%. In vitro, we evaluated the characteristics of the scaffolds using the MC3T3-E1 preosteoblasts over a 14 day period. In vitro characterization involved the use of the Alamar blue assay, confocal imaging, and real-time quantitative polymerase chain reaction. The results of this study demonstrate the potential of 3D printed biocomposite scaffolds, consisting of thermoplastic PCL reinforced with ESPs, as a promising alternative for bone-graft applications.

The present investigation was aimed at predicting a still (i.e., dead) vs. live embryo within a hatching goose egg by measuring the eggshell cooling rate. For this, we daily measured the temperature (T) values on the shell surface of goose eggs after they were removed from the incubator and during further natural cooling. T was recorded every 0.5 h for further 1.5 h of cooling. It was possible to recognize eggs with dead embryos using the combination of T, egg weight (W), and surface area (S). The resultant indicator (TS/W) was called specific temperature index (STI). The mathematical relationship using STI measurements between Days 8-13 facilitated 80 % correct identification of the eggs with dead embryos. Additionally, we derived mathematical dependencies for shell weight (Ws) and thickness (t) by utilizing the values of W, egg volume (V), S, the average T of all measurements taken, as well as the drop in T during 1.5 h of natural cooling. The key advantage of these parameters was their measurement and/or calculation by applying non-destructive methods. The integrated application of these parameters resulted in achieving high calculation accuracy as judged by correlation coefficients of 0.908 for Ws and 0.593 for t. These novel mathematical models have the potential to decrease hatching waste by predicting embryo viability. Our research will add to a toolkit for non-invasive egg assessment that is useful in the poultry industry, research on eggs, and engineering.

This study investigated the effects of different doses of limestone, light durations, light intensities, and vitamins on both the productive performance and egg quality. The study utilized two rearing houses (control and treatment), each accommodating 75000 Lohmann Brown Classic chicks reared in open-sided rearing cages from one day old until they reached 89 weeks of age. Throughout the laying period, the hens were subjected to a specific light regimen (light = 14 h; dark = 10 h a day). At the end of experiment, the treatment group displayed significant (p<0.05) differences compared to the control group across various parameters. Notably, the treatment group exhibited lower daily feed intake (treatment: 112 g/bird vs control: 115 g/bird), 9.6% higher egg production (treatment: 78.5% vs control: 68.9%), lower body weight (treatment: 2057 g vs control: 2073 g), lower feed conversion ratio (FCR)/egg (treatment: 1.44 vs control: 1.69), higher egg weight (treatment: 69.4 g vs control: 68.5 g), greater egg mass (treatment: 56.14 vs control: 48.76), greater shell thickness (treatment: 3.52 mm vs control: 3.44 mm), and greater shell weight (treatment: 9.3 g vs control: 8.79 g). However, the albumin weight, yolk weight, yolk diameter, shape index, and Haugh units (HU) were not significantly (p˃0.05) affected after 75 weeks of treatment when compared with those of the control group. Therefore, this study is the first of its kind to demonstrate that different ratios of limestone, different durations and intensities of light, and different vitamin supplementation doses in the treatment group (subjected to the novel rearing recommendations described in this study) may yield a profit of 180,541 USD, exceeding the baseline profit of the control group (subjected to conventional rearing methods).