Chitosan-treated Groups Research Articles

Efficacy of liver free and Chitosan against Eimeria tenella in chickens

Eimeria spp. are the pathogen that causes coccidiosis, a significant disease that affects intensively reared livestock, especially poultry. Anticoccidial feed additives, chemicals, and ionophores have routinely been employed to reduce Eimeria infections in broiler production. Therefore, the shift to antibiotic-free and organic farming necessitates novel coccidiosis preventive strategies. The present study evaluated the effects of potential feed additives, liver free and chitosan, against Eimeria tenella infection in White Leghorn broiler female chickens. One hundred sixty-five 1-day-old White Leghorn broiler female chicks were divided into 11 groups (15 female chicks per group), including the positive control group (G1), the negative control group (G2), a chitosan-treated group (G3), a chitosan-treated-infected group (G4), the liver free-treated group (G5), the liver free-treated-infected group (G6), the liver free-and-chitosan-treated group (G7), the liver free-and-chitosan-infected group (G8), the therapeutic liver free-and-chitosan-treated-infected group (G9), the sulfaquinoxaline-treated group (G10), and the sulfaquinoxaline-treated-infected group (G11). Chitosan was fed to the chicks in G3 and G4 as a preventative measure at a dose of 250 mg/kg. The G5 and G6 groups received 1.5 mg/kg of Liverfree. The G7 and G8 groups received chitosan and Liverfree. The G10 and G11 groups were administered 2 g/L of sulfaquinoxaline. From the moment the chicks arrived at Foshan University (one-day-old chicks) until the completion of the experiment, all medications were given to them as a preventative measure. G8 did; however, receive chitosan and liver free as therapeutic supplements at 7 dpi. The current study showed that the combination of liver free and chitosan can achieve better prophylactic and therapeutic effects than either alone. In E. tenella challenged chickens, G8 and G9 chickens showed reduced oocyst shedding and lesion score, improved growth performance (body weight, body weight gain, feed intake, feed conversion ratio, and mortality rate), and cecal histology. The current study demonstrates that combining liver free and chitosan has superior preventive and therapeutic benefits than either alone, and they could also be used as alternative anticoccidial agents.

Effects of Shrimp Shell-Derived Chitosan on Growth, Immunity, Intestinal Morphology, and Gene Expression of Nile Tilapia (Oreochromis niloticus) Reared in a Biofloc System.

Chitosan (CH) shows great potential as an immunostimulatory feed additive in aquaculture. This study evaluates the effects of varying dietary CH levels on the growth, immunity, intestinal morphology, and antioxidant status of Nile tilapia (Oreochromis niloticus) reared in a biofloc system. Tilapia fingerlings (mean weight 13.54 ± 0.05 g) were fed diets supplemented with 0 (CH0), 5 (CH5), 10 (CH10), 20 (CH20), and 40 (CH40) mL·kg-1 of CH for 8 weeks. Parameters were assessed after 4 and 8 weeks. Their final weight was not affected by CH supplementation, but CH at 10 mL·kg-1 significantly improved weight gain (WG) and specific growth rate (SGR) compared to the control (p < 0.05) at 8 weeks. Skin mucus lysozyme and peroxidase activities were lower in the chitosan-treated groups at weeks 4 and 8. Intestinal villi length and width were enhanced by 10 and 20 mL·kg-1 CH compared to the control. However, 40 mL·kg-1 CH caused detrimental impacts on the villi and muscular layer. CH supplementation, especially 5-10 mL·kg-1, increased liver and intestinal expressions of interleukin 1 (IL-1), interleukin 8 (IL-8), LPS-binding protein (LBP), glutathione reductase (GSR), glutathione peroxidase (GPX), and glutathione S-transferase (GST-α) compared to the control group. Overall, dietary CH at 10 mL·kg-1 can effectively promote growth, intestinal morphology, innate immunity, and antioxidant capacity in Nile tilapia fingerlings reared in biofloc systems.

Regeneration of olfactory neuroepithelium in 3-methylindole-induced anosmic rats treated with intranasal chitosan

Olfactory dysfunction significantly impairs the life quality of patients but without effective treatments to date. The previous report has demonstrated that chitosan mediates the differentiation of olfactory receptor neurons (ORNs) through insulin-like growth factors and insulin-like growth factor binding protein-2 axis in an in vitro model. However, whether chitosan can further treat olfactory dysfunction in vivo remains unexplored. This study aims to evaluate the therapeutic effect of chitosan on a 3-methylindole-induced anosmic rat model. Intraperitoneal injection of 3-methylindole is performed to induce anosmia in rats. Experimental results demonstrate that the food-finding duration after chitosan treatment gradually decrease to around 80 s, and both the olfactory neuroepithelium (ON) thickness and mature ORNs (expressing olfactory marker protein) are significantly restored. Furthermore, proliferating cells (expressing bromodeoxyuridine) are mainly co-expressed with immature ORNs (expressing βIII tubulin) below the intermediate layer of the ON in the chitosan-treated group on day 28 following 3-methylindole treatment. Conversely, proliferating cells are scattered over the ON, and co-localized with immature ORNs and sustentacular cells (expressing keratin 18) in the sham group, and even immature ORNs go into apoptosis (expressing DNA fragmentation and cleaved caspase-3), possibly causing incomplete regeneration. Consequently, chitosan regenerates the ON by regulating olfactory neural homeostasis and reducing ORN apoptosis, and serves as a potential therapeutic intervention for olfactory dysfunction in the future.

Impact of onboard chitosan treatment of whole cod (Gadus morhua) on the shelf life and spoilage bacteria of loins stored superchilled under different atmospheres

The initial handling of marine fish on board fishing vessels is crucial to retain freshness and ensure an extended shelf life of the resulting fresh products. Here the effect of onboard chitosan treatment of whole, gutted Atlantic cod (Gadus morhua) was studied by evaluating the quality and shelf life of loins processed six days post-catch and packaged in air or modified atmosphere (% CO2/O2/N2: 55/5/40) and stored superchilled for 11 and 16 days, respectively. Sensory evaluation did not reveal a clear effect of chitosan treatment on sensory characteristics, length of freshness period or shelf life of loins under either packaging conditions throughout the storage period. However, directly after loin processing, microbiological analysis of loins showed that onboard chitosan treatment led to significantly lower total viable counts as well as lower counts of specific spoilage organisms (SSO), such as H2S-producers and Pseudomonas spp., compared to the untreated group. In addition, the culture-independent approach revealed a lower bacterial diversity in the chitosan-treated groups compared to the untreated groups, independently of packaging method. Partial 16S rRNA gene sequences belonging to Photobacterium dominated all sample groups, indicating that this genus was likely the main contributor to the spoilage process.

Marine-Derived Chitosan Nanoparticles Improved the Intestinal Histo-Morphometrical Features in Association with the Health and Immune Response of Grey Mullet (Liza ramada).

Marine-derived substances are known for their beneficial influences on aquatic animals’ performances and are recommended to improve intestinal health, immunity, and anti-oxidative status. The present study investigates the role of chitosan nanoparticles on the intestinal histo-morphometrical features in association with the health and immune response of Grey Mullet (Liza ramada). Chitosan nanoparticles are included in the diets at 0, 0.5, 1, and 2 g/kg and introduced to fish in a successive feeding trial for eight weeks. The final body weight (FBW), weight gain (WG), and specific growth rate (SGR) parameters are significantly increased while feed conversion ratio (FCR) decreases by chitosan nanoparticles compared to the control (p < 0.05). The morphometric analysis of the intestines reveals a significant improvement in villus height, villus width, and the number of goblet cells in chitosan-treated groups in a dose-dependent manner. Additionally, there is a positive correlation between the thickness of the enterocyte brush border and the chitosan dose, referring to an increasing absorptive activity. Histologically, the intestinal wall of Grey Mullet consists of four layers; mucosa, sub-mucosa, tunica muscularis (muscular layers), and serosa. The histological examination of the L. ramada intestine shows a normal histo-morphology. The epithelial layer of intestinal mucosa is thrown into elongated finger-like projections, the intestinal villi. The values of hemoglobin, hematocrit, red blood cells (RBCs), total protein (TP), albumin, and globulin are significantly increased in fish fed 1, and 2 g/kg of chitosan nanoparticles compared to fish fed 0 and 0.5 g/kg (p < 0.05). The highest levels of TP and albumin are observed in fish fed 1 g/kg diet (p < 0.05). The lysozyme activity and phagocytic index are significantly enhanced by feeding chitosan nanoparticles at 0.5, 1, and 2 g/kg, whereas the phagocytic activity is improved in fish fed 1 and 2 g/kg (p < 0.05). The highest lysozyme activity and phagocytic index are observed in fish fed 1 g/kg. SOD is significantly activated by feeding chitosan nanoparticles at 1 g/kg. Simultaneously, glutathione peroxidase (GPx) and catalase (CAT) activities also are enhanced by feeding chitosan at 1 and 2 g/kg, compared to fish fed 0 and 0.5 g/kg (p < 0.05). The highest GPx and CAT activities are observed in fish fed 1 g/kg (p < 0.05). Conversely, the malondialdehyde (MDA) levels are decreased by feeding chitosan at 1 and 2 g/kg, with the lowest being in fish fed 1 g/kg (p < 0.05). To summarize, the results elucidate that L. ramada fed dietary chitosan nanoparticles have a marked growth rate, immune response, and anti-oxidative response. These improvements are attributed to the potential role of chitosan nanoparticles in enhancing intestinal histo-morphometry and intestinal health. These results soundly support the possibility of using chitosan nanoparticles at 1–2 g/kg as a feasible functional supplement for aquatic animals.

Biomimetic chitosan against bioinspired nanohydroxyapatite for repairing enamel surfaces

In this study, chitosan was employed as a novel biomimetic mineralization model to repair damaged enamel to compare its performance with that of bioinspired zinc-doped nanohydroxyapatite. Fifty human premolar tooth slices were prepared, and artificial caries lesions were induced to produce demineralized enamel surfaces. The etched slices were randomly divided into two groups: a chitosan-hydrogel-treated group and a zinc-doped nanohydroxyapatite-treated group. In vitro assessment using energy-dispersive X-ray analysis, X-ray diffraction analysis and scanning electron microscopy was conducted at the baseline, demineralization and remineralization stages. Baseline results were matched with those for normal enamel; a marked reduction in the calcium (Ca)/phosphorus (P) ratio to 1·12 and the lack of the characteristic hydroxyapatite diffraction peaks were detected for demineralized enamel. The remineralization stage revealed evident recovery of the mineral contents (the calcium/phosphorus ratio was 1·61 for the chitosan-treated group and 1·58 for the bioinspired-nanohydroxyapatite-treated one), with apparent distinctive X-ray diffraction patterns of hydroxyapatite in both groups. Scanning electron microscopic analysis showed the absence of etched enamel porosity, with the formation of a newly formed rod-like apatite layer, similar to natural enamel, which extended over the treated enamel surfaces of both groups. Chitosan hydrogel is recommended as a biomimetic mineralization smart system for repairing demineralized carious enamel.

Bone Formation Using Cross-Linked Chitosan Scaffolds in Rat Calvarial Defects.

To investigate the osteoconductive effect of a chitosan scaffold in a rat skull defect model. Previous publications have demonstrated the osteoinductive properties as scaffold materials with growth factors; however, whether chitosan alone has osteoconductive ability is unclear. This study used cross-linked chitosan scaffolds for in vivo evaluation of scaffold-supported bone regeneration in rat calvarial defects using histopathological analysis and examination of alkaline phosphatase (ALP), calcium, phosphorus, and calcitonin serum levels. Scaffolds were made of cross-linked chitosan. After the defect was filled with the scaffold, the periosteum was carefully repositioned and sutured to stabilize the scaffold. The effects of the scaffold on wound repair were examined microscopically. Morphological radiographic and histopathological analyses of wound repair ratios were performed at 3 and 4 weeks after the defects were made. Using the cross-linked chitosan biomaterial of the wounds. The amount of regenerated bone measured was significantly greater in the chitosan-treated group than in the control group. The ALP level in the chitosan group at 4 weeks was higher than at baseline and at the 4-week follow-up in the control group (P < 0.05). The results show that cross-linked chitosan has an osteoconductive effect on bone regeneration in vivo.

The sinomenine enteric-coated microspheres suppressed the TLR/NF-κB signaling in DSS-induced experimental colitis

Sinomenine is a pure alkaloid with immunosuppressive effects that is extracted from the Chinese medicinal plant Sinomenium acutum. We studied the therapeutic effects of sinomenine on inflammatory bowel disease. In this study, we randomly divided mice into the following ten groups: Control group; DSS-induced colitis group; Salicylazosulfapyridine (SASP)-treated group; Chitosan-treated group; low-, medium-, and high-dose sinomenine-treated and sinomenine enteric-coated microspheres-treated groups. We recorded changes in colon length, disease activity index (DAI), and colon pathology, measured TLR4, MyD88, SIGIRR, NF-κB p65 protein levels and inflammatory serum cytokine levels.Except for the Control group, the weight of mice in each group decreased, the DAI of the DSS-induced colitis group was significantly higher than the other groups, and the DAIs of the sinomenine- and sinomenine enteric-coated microspheres-treated groups were significantly lower than that of the SASP-treated group. TLR4, MyD88, NF-κB p65 and proinflammatory cytokine expressions decreased dose dependently in the sinomenine and sinomenine enteric-coated microspheres-treated groups and were generally lower in the sinomenine enteric-coated microspheres groups. However, SIGIRR and anti-inflammatory IL-10 expressions exhibited the opposite pattern. Based on the superior therapeutic effect, sinomenine enteric-coated microspheres might regulate TLR/NF-κB signaling and would be beneficial for an effective and safe therapy of inflammatory bowel disease.

Application of platelet derived growth factor-BB and diabetic wound healing: the relationship with oxidative events

The reasons that cause delay in wound healing in diabetes are a decrease in the level of growth factors secretion, an increase in the destruction of growth factors and in oxidative stress. Platelet derived growth factor (PDGF) is one of the important growth factors that play a role in all phases of wound healing. This study investigates time-dependent effects of topically PDGF-BB administration on oxidative events on the healing of dorsolateral-excisional wounds in diabetic rats. Forty-two female Wistar-albino rats with streptozotocin-induced diabetes were divided into four groups: control group, untreated group, chitosan-treated group, chitosan + PDGF-BB-treated group. Two identical full-thickness excisional skin wounds were made under anaesthesia in all rats except for the control group. In the PDGF-BB-treated and chitosan-treated groups, the wounds were treated topically PDGF-BB (7 ng/mL, single daily dose) and blank chitosan gel (equal amount) after wounding, respectively. After these administrations, on day 3 and day 7 of wound healing, rats were sacrificed. Thiobarbituric acid reactive substances, glutathione, nitric oxide, ascorbic acid levels, and superoxide dismutase activity in wound tissues were spectrophotometrically measured. PDGF-BB administration significantly increased TBARS levels and non-enzymatic antioxidant levels in early phase of diabetic wound healing. PDGF-BB dramatically reduced NOx levels on day 3 and sharply increased NOx levels on day 7 of wound healing. Consequently, PDGF-BB administration can be effective on oxidative balance in the early phase of diabetic wound healing.

Synergistic in vitro photodynamic antimicrobial activity of methylene blue and chitosan against Helicobacter pylori 26695

Photodynamic therapy (PDT) is a method for killing cells (bacterial, fungal and cancer cells) or virus using photosensitizers (PS) and light of various wavelengths. In vitro PDT using endoscopic light against H. pylori was effective at a concentration of 0.2mg/mL of MB. The purpose of this study was to increase the effect of photodynamic modality against H. pylori by addition of chitosan to MB. The bactericidal effect was measured by counting viable cells after PDT. The degree of damage to DNA was confirmed using alkaline gel electrophoresis. Cellular DNA damage was demonstrated by ethidium bromide monoazide-quantitative polymerase chain reaction (EMA-qPCR). In the groups treated with either 0.04 mg/mL MB alone or 0.02 mg/mL MB with endoscopic light for 15 min, viable cells were decreased approximately tenfold. The group treated with 0.04 mg/mL of MB with light, showed more effective bactericidal activity than 0.02 mg/mL of MB treatment. By 0.05% chitosan pre-treatment followed with 0.04 mg/mL of MB and light irradiation, viable cells were decreased 10(7)-fold. The DNA damage caused by PDT as demonstrated by alkaline gel electrophoresis was greater in the MB plus chitosan-treated group than in control and MB-treated groups. Cellular DNA damage demonstrated by EMA-qPCR was also greater in the group treated with MB plus chitosan than in the MB-treated group. The bactericidal effects with PDT using MB were increased with the concentration of photosensitizer and chitosan treatment, peculiarly before endoscopic light irradiation.

The effect of chitosan on the concentration of 17β-estradiol and free triiodothyronine in mice exposed to polychlorinated biphenyls (PCBs)

The aim of this work was to examine (i) how the applied PCB mixture influences the level of 17β-estradiol (E2) and free triiodothyronine (FT3) in the blood plasma of mice (C57/BL/6J) and (ii) whether supplementation with chitosan would protect against the observed changes in the examined plasma hormone concentrations. In the study we used a mixture of indicator PCBs (CB no. 28, 52, 101, 118, 138, 153, 180) and our results showed their anti-estrogenic effects. Exposure to the mixture resulted in a significant decrease (P < 0.05) in plasma concentration of E2 relative to the control, and chitosan administration did not prevent the decrease. To the contrary, E2 concentration in the blood plasma of the mice which received both the PCB mixture and chitosan was lower compared to those which did not receive chitosan. Exposure to the PCBs also resulted in a decrease in FT3 concentration in the treatedgroup, although it was not as pronounced as for E2 and was prevented with dietary supplementation with chitosan, with the observed FT3 level in the chitosan-treated group similar to the control. In summary, supplementation with chitosan can only to a certain extent minimize the negative effects of exposure to PCBs.

Inhibitory effect of chitosan-mediated cytokine response modifier A on interkin-1β and tumor necrosis factor-α expression of chondrocytes

Objective To investigate the effects of chitosan/pCDNA3.1 + cytokine rcsponse modifier A (CrmA) nanoparticles on interkin-1β (IL-1β) and tumor necrosis factor-α (TNF-α) expression of chondrocytes.Methods Rabbit chondrocytes were isolated and cultured,and treated with PBS,10 mg/L chitosan (CS) or chitosan/pCDNA3.1 + CrmA nanoparticles,respectively for 6 h.Then 10 μ g/L IL-1β was added into the culture medium.After 48 h,the mRNA expression of IL-13 and TNF-α in chondrocytes was detected by using real-time polymerase chain reaction.Results In chitosan/pCDNA3.1 + CrmA-treated group (0.55 ±0.08),the mRNA expression of IL-1β in chondrocytes was significantly suppressed as compared with corresponding samples of chitosan-treated group (0.69 ± 0.06,P ＜ 0.01) and PBS-treated group (0.99 ±0.04,P ＜ 0.01).There was significant difference in the I L-1 β expression between chitosantreated group and PBS-treated group (P ＜ 0.01).The TNF-α mRNA expression of chondrocytes in chitosan/pCDNA3.1 + CrmA-treated group (0.57 ± 0.07) was lower than that in chitosan-treated group (0.71 ± 0.05,P ＜ 0.01) and PBS-treated group (0.99 ± 0.06,P ＜ 0.01).Significant difference of TNF-α expression between chitoson treated group and PBS treated group was observed (P ＜ 0.01).Conclusion CrmA mediated by chitosan could significantly suppress the mRNA expression of IL-1β and TNF-α of chondrocytes induced bv interkin-1 β.chitosan/pCDNA3.1 * CrmA nanoparticles maybe a potential candidate for therapy of osteoarthritis. Key words: Osteoarthritis; Chondrocyte; Chitosan; Cytokine response modifier A

PDGF supplementation alters oxidative events in wound healing process: a time course study

Platelet-derived growth factor (PDGF), an important stimulant, plays a role in almost all stages of wound healing process. In various studies, it has been shown that PDGF has healing effects in this process. In the present study, we especially focused on investigating the effects of exogenous PDGF administration on oxidative events during cutaneous wound healing process. Experiments were performed on 42 female Wistar-albino rats. Animals were divided into four groups: control, untreated, chitosan-treated and chitosan + PDGF-treated. Two uniform full-thickness excisional skin wounds were made under anesthesia in all animals except control group. In the chitosan + PDGF-treated groups, the wounds were treated topically with a single daily dose PDGF-BB (7 ng/ml) after wounding. In the chitosan-treated groups, the wounds were treated topically with equal amount of blank chitosan gel. After that, on the 3rd and 7th days of wound healing, the animals were killed. Thiobarbituric acid-reactive substances (TBARS), nitric oxide (NOx), ascorbic acid (AA), glutathione (GSH) levels and superoxide dismutase (SOD) activity were measured spectrophotometrically in the wound tissues. PDGF significantly increased TBARS levels in early phase of wound healing. In contrast, it significantly decreased TBARS levels in later phase of healing. In the chitosan + PDGF-treated group, NOx levels decreased on days 3 and 7 when compared with the chitosan-treated groups. Non-enzymatic antioxidant levels were increased by PDGF administration and this may have contributed to increase in wound tissue antioxidant capacity. In the light of these findings, PDGF supplementation may have altering effects on oxidative events depending on the time in wound healing process.

키토산의 급여량과 급여 방법에 따른 육계의 성장 및 닭 다리육의 품질에 미치는 영향

This study was conducted to determine the effects of dietary supplementation with chitosan feeding levels and feeding forms on the performance, proximate composition, pH, thiobarbituric acid reactive substance (TBARS), water holding capacity (WHC), shear force, meat color, and fatty acid of chicken thigh meat. Two hundred broilers (Arbor Acre Broiler, male) were randomly assigned to five groups and were fed for five weeks and slaughtered. Thigh muscle was evaluated in this experiment. The amounts of proximate composition, crude fat of control and T3 for the chitosan-treated groups were significantly higher as compared with T2 (P<0.05), but no significant effects were detected on moisture, crude protein, and crude ash. By comparison, pH was significantly increased when chitosan was included at 2% into diet or more than 1% into drinking water. TBARS was significantly lower at chitosan treated groups it was decreased with increasing chitosan level in the diets (P<0.05). Therefore, chitosan had the possibility to improve shelf life of chicken meat. Higher chitosan levels and feeding additive increased high density lipoprotein cholesterol (HDL-C) and reduced total cholesterol and low density lipoprotein cholesterol (LDL-C) (P<0.05). L * and a * of chitosan treated group was higher than the control. As the dietary chitosan levels increase, the composition of palmitic acid and oleic acid levels was increased, however, those but the rates of linoleic acid and arachidonic acid were decreased. It is concluded that dietary chitosan has a positive effect on increasing HDL-C and oleic acid and decreasing total-C, LDL-C and TBARS values. Therefore, the treatment with the most significant effects in the current study was the high level of chitosan.

Antibacterial Activities of Chitosans and Chitosan Oligomers with Different Molecular Weights on Spoilage Bacteria Isolated from Tofu

ABSTRACTSeven bacteria were isolated from spoiled tofu and identified as Bacillus sp. (S08), B. megaterium (S10), B. cereus (S17, S27, S28, S32), and Enterobacter sakazakii (S35). In a paper disc test with 6 chitosans and 6 chitosan oligomers of different molecular weights, chitosans showed higher antimicrobial activity than did chitosan oligomers at a 0.1% concentration. Results of inhibitory effects of 6 chitosans on growth of Bacillus sp. (S08) failed to detect viable cells after incubation for 24 hrs at 37 C, even at 0.02% concentration. With B. megaterium (S10) and B. cereus (S27), a 3 to 4 log cycle reduction was found in the chitosan‐treated group. The growth of Enterobacter sakazakii (S35) was completely suppressed in the presence of 0.04% chitosan except for 1 chitosan product. The minimum inhibitory concentration of chitosan differed with products and isolates, ranging from 0.005% to above 0.1%.