BACKGROUND: Numerous studies have investigated the positive effect of chitosan and nano-chitosan loaded clinoptilolite on growth performance, digestive enzyme activity, and intestinal histomorphology in different fish species. However, there are no data evaluating the potential effect of the composites on the fish stomach.OBJECTIVES: In the current study, the effects of chitosan and nano-chitosan loaded clinoptilolite on histological features and pepsin activity in the rainbow trout stomach were considered.METHODS: Chitosan and nano-chitosan loaded clinoptilolite were synthesized, and then two hundred and forty fish (~27.75 g) fish were distributed in eight groups each in three replicates. Ten days after adaptation, the fish were fed with eight diets, including control diet (CTR), clinpotilolite (T1), chitosan composites (T2, T3, T4), and nano-chitosan composites (T5, T6, T7) for 70 days. Afterward, all fish in each tank were anesthetized in clove extract (50 μl/l), and tissue samples were obtained for pepsin activity (n= 5) and histological assay (n = 5).RESULTS: The groups administrated with nanochitosan composites showed the highest pepsin activity (P˂0.05). Additionally, histological examinations exhibited a higher epithelial height, increased mucosal density, and oxynticopeptic cells hypertrophy in fish fed composites compared to the CTR group (P˂0.05). Meanwhile, nanochitosan composite administration could cause higher reaction of secreted granules to periodic acid–Schiff (PAS) staining.CONCLUSIONS: The findings demonstrated the potential application of chitosan and nano-chitosan loaded clinoptilolite composites for improvements in the histomorphology and pepsin activity of the rainbow trout stomach, resulting in higher growth performance.

BACKGROUND: Probiotics have more functional effects and less survival  under hard acidic-bile circumstances of digestive system, and foodstuff products situation has persuaded investigators to find techniques to resolve this problem. Microencapsulation as a useful method has a perceptible effect in this regard. Objectives: The aim of this study was to assess the protective role of double coated beads of calcium alginate-chitosan-eudragit S100 achieved from microencapsulation of Lactobacillus acidophilus as a predominant flora of human and animals gut. Methods: Following activation of starter culture of L.acidophilus in MRS-broth medium,  centrifuge (at aspeed of 5000 rpm for 10 minutes) was used to purify bacteria. Extrusion technique was used for Microencapsulation of probiotic bacterium. The survey of beads solidity was carried out for 12 hours and the study of survival of microencapsulated bacteria was done for 120 minutes inside hydrochloric acid, phosphate buffer and digestive powder solution.  MRS-Salicin-agar and pour plate method and incubation at 37oC for 48 h was done for cultivation. Data were analyzed by means of an independent t-test. Results: Shape and size of beads were shown by optical microscope. The consequences demonstrated that survivability of microencapsulated bacteria in the mentioned conditions, in both situation with and without mechanical tensions, is significantly more than free bacteria (p<0/05). Conclusions: Microencapsulation with calcium alginate- chitosan-eudragit S100 plays a significant role  in increasing the rate of L. acidophilus viability.

BACKGROUND: Poultry meat belongs to perishable foods and the major concern of food industries is the microbial spoilage of poultry meat. OBJECTIVES: The aim of present study was to investigate the effect of chitosan (CH) coating enriched with different concentrations of Ziziphora clinopodioides essential oil (ZEO) in comparison with control group on some of the sensory and microbial properties of chicken breast fillets during storage at refrigerated temperature for 12 days.   METHODS: Essential oil extraction was done by hydro-distillation method and its chemical composition was determined by gas chromatography with mass spectrometry (GC-MS). In the present study, chicken breast fillets separately were dipped in 2% CH solution containing ZEO at concentrations 0, 0.5 and 1% and then stored at refrigerated condition for 12 days. After that chicken fillets were studied at 7 intervals (0, 1, 3, 5, 7, 9 and 12 days) regarding microbial (Total mesophilic and Psychrotrophic bacteria, Pseudomonas spp. and Enterobacteriaceae) and sensory (color, odor and taste) examination. RESULTS: The most important compounds of the ZEO were geraniol (20.62%), carvacrol (18.17%), thymol (5.39%), α-terpineol (7.49%) and 4-terpineol (6.83%). Results of this study revealed that in the treatments coated with CH containing ZEO, total viable counts (TVC), Pseudomonas spp., lactic acid bacteria, Psychrotrophic bacteria and Enterobacteriaceae familysignificantly (P<0.05) decreased as compared to control group during the storage period. Based on the results of the present study, coating of chicken fillets with chitosan alone or chitosan containing 0.5 % concentration of ZEO showed better sensory properties. CONCLUSIONS: CH coating enriched with 0.5 % ZEO has potential to extend shelf life of chicken fillets without any unfavorable organoleptic properties.

BACKGROUND: Many herbs such as fenugreek and orange have compounds with antioxidant properties, which can increase the shelf life of foods. Considering that fish are a popular food among consumers, they are susceptible to rapid corruption. OBJECTIVES: In this study, to improve rainbow trout (Oncorhynchus mykiss) fillet shelf life at refrigerated condition, orange juice concentrate and chitosan coating enriched with Fenugreek (Trigonella foenum-graecum) essential oil was used. METHODS: In the present study, 8 treatments were evaluated for 12 days at refrigerator temperature for Chemical indicators PH value, Total volatile nitrogen (TVN), Thiobarbituric Acid (TBA), Peroxide value (PV), Free Fatty Acid (FFA), and Sensory Properties. RESULTS: According to statistical results, fish fillets coated with chitosan enriched with 2% fenugreek essential oil and immersed in orange juice concentrate were lower than other groups for all chemical indicators and had a significant difference with the control group (P<0.05). In the DPPH test, the most inhibitory effect after BHT (butylated hydroxy toluene) was orange juice with 1.39 brix and then black pepper 2%, respectively. In the RP test, the absorbance of the coated sample with chitosan containing fenugreek essential oil and orange juice concentrate with BHT did not show any significant difference (P>0.05). Sensory evaluation also showed that the chitosan-coated sample containing fenugreek essential oil and orange juice concentrate improved the sensory index during storage compared to other groups, especially the control group. CONCLUSIONS: According to the results, it can be stated that the use of orange juice concentrate, chitosan coating and fenugreek essential oil have a significant effect on reducing the oxidation process of rainbow trout fillet at refrigerator temperature.

BACKGROUND: Many drugs such as atorvastatin are known effective in reduction of serum lipids in dogs, but with a literature review, we did not find report on the field of the effect of chitosan on serum lipid in dogs. OBJECTIVES: The purpose of the present survey was comparative evaluation of the effects of chitosan and atorvastatin on serum lipid profile changes and the influence of time on treatment process in dogs. METHODS: For the management of cholesterol powder induced hyperlipidemia, twenty healthy dogs were randomly divided into four equal groups. Group A (control) included of five dogs that were fed with cholesterol powder (4 gr/kg for 10 days). Group B was similar to group A, but in addition, atorvastatin (5 mg/kg) was administered for 45 days after induced hyperlipidemia. Group C was similar to group B, but chitosan (3 gr/dog) was administered instead of atorvastatin. Group D was a combination of groups B and C, which the combination of atorvastatin and chitosan were fed to dogs with the same dose of previous groups. Blood samples were collected four times on days 0, 10, 40 and 55 after challenge, then serum triglycerides, total cholesterol, HDL-C and LDL-C levels were measured using standard commercial kits. RESULTS: Groups of atorvastatin and chitosan (B and C) and group D were more effective in lowering serum triglyceride, total cholesterol and LDL-C and increase of HDL-C, compared with group A (p<0.05). The greatest decrease was related to group D for triglyceride (105.60±17.49), total cholesterol (119.80±11.39) and LDL-C (36.40±7.57). The greatest increase was seen in group D for HDL-C (36.40±7.57) also. In comparison between two drugs and their effects on lipid profiles, atorvastatin showed a significant difference than chitosan (p<0.05). A combination of two drugs, was more effective compared with single administration of the drugs (p<0.05). CONCLUSIONS: The present survey showed that although both drugs have hypolipidemic activity in dogs, but the effect of chitosan was lower than atorvastatin, so it is not recommended to use chitosan only. Further experimentation needs to elucidate the possible mechanism of the drugs.

Introduction: The development of Jembrana disease vaccine is an important effort to prevent losses in the Bali cattle industry in Indonesia. This study aims to prepare a Jembrana DNA vaccine encoding the transmembrane portion of the envelope protein in pEGFP-C1 and test the success of its delivery in culture cells using a chitosan-DNA complex. Material and Methods: Cloning of the DNA vaccine was successfully performed on E. coli DH5α and confirmed by colony PCR, restriction analysis and sequencing. The plasmids were prepared as a chitosan complex using the complex coacervation method and physicochemically characterised using a particle size analyser. A transfection assay was performed in HeLa cells with 4 h exposure, and mRNA expression was assessed at 24 h post transfection. Results: With a 1:2 (wt./wt.) ratio of DNA and chitosan, the complexes have a mean diameter of 236 nm, zeta potential value of + 17.9 mV, and showed no high toxicity potential in the HeLa cells. This complex successfully delivered the DNA into cells, as shown by the presence of a specific RT-PCR product (336 bp). However, the real-time PCR analysis showed that the delivery with chitosan complex resulted in lower target mRNA expression when compared with a commercial transfecting agent. Conclusion: pEGFP-env-tm JDV as a candidate vaccine can be delivered as the chitosan-DNA complex and be expressed at the transcription level in vitro. This initial study will be used for further improvement and evaluation in vivo.

Chitosan is similar in structure to cellulose and are the second most abundant polysaccharides in nature, comprising the horny substance in the exoskeletons of crabs, shrimp and insects as well as fungi. This study was conducted to access the effect of immunomodulation responses of chitosan(N-acetyl-β-Dglucosamine) chicken infected with in Fowl typhoid(Salmonella gallinarum). One-day-old broiler chicks were divided into eight groups: The 1st group was inoculated intra-peritoneally with chitosan and challenged intra-peritoneally with S. gallinarum. The 2nd group was inoculated intra-peritoneally with chitosan.

The development of safe, novel adjuvant is necessary to maximize the efficacy of new and/or available vaccines. In this study, three different molecular weights of chitosan (low, medium, high) were evaluated as immunopotentiators/adjuvants of inactivated rabies vaccine in white Norway rats as an experimental model. Two concentrations (1.5% & 3% w/v) of each chitosan type were used in final concentration 5 mg/ml and 10 mg/ml. Results showed that lymphocyte proliferation were significantly elevated (P < 0.05) in all chitosan vaccinated rats compared to aluminum hydroxide gel vaccinated one. All three type of chitosan produced accelerated and enhanced effects on rabies-neutralizing antibody responses in vaccinated groups. Increases in antibody titers together with lymphocytes proliferation responses revealed that chitosan induced both humoral and cell-mediated immune responses. When compared with aluminum hydroxide vaccine adjuvant, chitosan was superior to aluminum hydroxide. The conclusion of these results suggested that chitosan with different MW had a strong potential to increase both cellular and humoral immune responses and that chitosan may be a promising and efficacious adjuvant candidate suitable for inactivated rabies vaccine.

This study was conducted to compare the hemostatic efficacy of three ferric subsulfate- and chitosan-based styptics as a powder and a gel containing ferric subsulfate and chitosan (FSC-PO and FSC-G, respectively) and a soaked pad containing ferric subsulfate and lidocaine (FSL-SP) using a rat tail bleeding model. The cytotoxicity of the styptics against L-929 mouse fibroblasts was also evaluated using a cell counting kit-8 assay. Four groups of 10 rats each were assigned to the three different styptics and a non-treated control groups. Rat tail tips were transected, after which styptics were applied with pressure. The wounds were observed for hemostasis for 3 min, then irrigated with saline to check for recurrent hemorrhage. L-929 mouse fibroblasts were exposed to extracts of the styptics (100 mg/ mL) and their dilutions (1:10, 1:100, and 1:1,000). FSC-PO and FSC-G more effectively controlled initial hemorrhage than FSL-SP (p = 0.033). Additionally, FSC-PO and FSC-G more effectively maintained hemostasis than the control group (p = 0.02 and p 0.01, respectively). However, all styptics showed enhanced cytotoxicity against L-929 cells in a dose-dependent manner. Therefore, although FSC-PO and FSC-G would be recommended to control hemorrhage, the benefits of styptics must be balanced against the clinical significance of their cytotoxicity.

Objective: This study was conducted to investigate the chelating effect of silver nitrate (AgNO3) by chitosan on growth performances, hematological and biochemical parameters, and the histopathological structure of the liver and the kidney in broiler chicken. Materials and methods: A total of 192 day-old Cobb 500 strain chicks were randomly assigned to 3 treatments of 64 chicks each. Control group was fed on basal diet without supplement (R0) and the two others groups were fed on rations supplemented with 10 mg of unchelated (RAg) or chelated (RCs-Ag) AgNO3 per Kg of feed, respectively. Parameters that have been studied consisted of feed intake, weight gain, blood and serum biochemical, and histopathological analyses of liver and kidney. Results: Results revealed that chelation of AgNO3 by chitosan did not have any effect on growth performances and hematological parameters in chicken. However, chelated and unchelated AgNO3 increased the serum content in triglyceride, and cholesterol and decreased the serum content in creatinin, albumin and alanine aminotransferase (ALAT). Chelating AgNO3 with chitosan prevented and corrected the toxicity induced on the histological structure of liver and kidney. Conclusion: Chitosan can be used as a chelating agent to alleviate the harmful effects of AgNO3 as silver ion for poultry. [J Adv Vet Anim Res 2017; 4(2.000): 187-193]