Microbiome Changes in Layer Pullets Reared in Floor Pens along the Growth Period.

The gastrointestinal tract microbiome is essential for regulating nutrient absorption, gut immune function, and host growth and development. In the present study, we characterized the development of ileum and cecum microbiota in pullets throughout the rearing period, encompassing a period from the day of hatching to 18 weeks of age. The growth performance and intestinal microbiome (ileum and cecum) of pullets were analyzed at 1, 5, 11, and 18 weeks of age. The richness of the ileum and cecum bacterial communities (alpha diversity) was higher in pullets at 18 weeks of age than in those at 1 and 5 weeks of age. Microbiota from weeks 1, 5, 11, and 18 were distinctly grouped in a NMDS plot, representing beta diversity within the ileum. However, the results for cecum microbiota did not reveal evident separation among the different age groups in the weighted UniFrac. In conclusion, our findings demonstrate variations and diversification in ileum and cecum microbiota across different rearing stages in pullets. These insights have the potential to inform the development of nutritional strategies that promote gut health and contribute to the improved development of pullets.

A comparative study on feeding timing and additive types of broilers in a high-temperature environment.

Antioxidants such as vitamin C (VC) and green tea extract (GTE) have been reported to have various antioxidant functions and are used as one of the nutritional approaches to alleviate heat stress (HS) in chickens. However, studies on the feeding timing that can produce optimal effects have not been reported. In this study, the stress-relieving effect of VC and GTE addition timing was investigated in high-temperature broiler chickens. A total of 880 1-d-old male chickens were used, and the treatments were as follows: no feed additives provided, CON; VC 250 mg/kg added from 1 d, VC1; GTE 600 mg/kg added from 1 d, GTE1; VC 250 mg/kg added from 22 d, VC22; GTE 600 mg/kg added from 22 d, GTE22. The HS environment was provided for 2 wk from the 22 d and was set at 33 ± 1 °C, 55 ± 10% for 24 h. Feed and water were provided ad libitum. Broiler production was similar in all treatments. In chicken meat quality, the addition of VC and GTE had an effect on meat color and pH (P < 0.05). In particular, GTE had a positive effect on the antioxidant capacity and quality preservation of breast meat (P < 0.05). In blood characteristics, GTE1 significantly lowered the level of total cholesterol, and VC1 affected AST and IgM (P < 0.05). Interestingly, the VC1 group had a positive effect on the maintenance and development of intestinal morphology, a lower rectal temperature, and showed to relieve stress. In conclusion, the addition of VC and GTE has been shown to alleviate the high-temperature stress of broilers, and in the case of VC in particular, feeding from 1 d appeared to alleviate stress more effectively. This study suggests that it is important to determine the appropriate timing of addition of functional substances in order to effectively reduce various stresses that occur in livestock rearing.

The increasing frequency of exposure to high-temperature environments has prompted research into nutritional approaches to alleviate heat stress in chickens, but little research has been reported on feeding timing. The aim of this study was to determine the effect of feeding timing on the effectiveness of the natural antioxidants vitamin C (VC) and green tea extract (GTE). Production was similar among all treatments, and GTE fed from 1 d of age increased antioxidant capacity, including DPPH, FRAP, and MDA in carcass quality. VC fed from 1 d of age decreased AST and increased IgM in the blood, and increased villus height (VH), with a positive effect on intestinal development. In conclusion, feeding VC and GTE from 1 d of age has been shown to effectively alleviate stress by increasing antioxidant capacity in breast meat, positively changing total cholesterol, AST, and IgM in the blood, and maintaining intestinal morphology, and it is important to set the timing of feeding to increase the effectiveness of the additives.

The potential of non-movement behavior observation method for detection of sick broiler chickens.

The poultry industry, which produces excellent sources of protein, suffers enormous economic damage from diseases. To solve this problem, research is being conducted on the early detection of infection according to the behavioral characteristics of poultry. The purpose of this study was to evaluate the potential of a non-movement behavior observation method to detect sick chickens. Forty 1-day-old Ross 308 males were used in the experiments, and an isolator equipped with an Internet Protocol (IP) camera was fabricated for observation. The chickens were inoculated with Salmonella enterica serovar Gallinarum A18-GCVP-014, the causative agent of fowl typhoid (FT), at 14 days of age, which is a vulnerable period for FT infection. The chickens were continuously observed with an IP camera for 2 weeks after inoculation, chickens that did not move for more than 30 minutes were detected and marked according to the algorithm. FT infection was confirmed based on clinical symptoms, analysis of cardiac, spleen and liver lesion scores, pathogen re-isolation, and serological analysis. As a result, clinical symptoms were first observed four days after inoculation, and dead chickens were observed on day six. Eleven days after inoculation, the number of clinical symptoms gradually decreased, indicating a state of recovery. For lesion scores, dead chickens scored 3.57 and live chickens scored 2.38. Pathogens were re-isolated in 37 out of 40 chickens, and hemagglutination test was positive in seven out of 26 chickens. The IP camera applied with the algorithm detected about 83% of the chickens that died in advance through non-movement behavior observation. Therefore, observation of non-movement behavior is one of the ways to detect infected chickens in advance, and it appears to have potential for the development of remote broiler management system.

Evaluation of Non-Fermented and Fermented Chinese Chive Juice as an Alternative to Antibiotic Growth Promoters of Broilers.

The present study explores the application of CC juice as a suitable feed additive and alternative to conventional antibiotics. We performed a comparative study to investigate the effects of non-fermented and fermented CC juice on broiler productivity, meat quality, blood characteristics, intestinal characteristics, and microbiota associated with intestinal characteristics. A total of 800 one-day-old Ross 308 broiler chickens were randomly assigned to one of the four dietary treatment groups: (1) basal diet (negative control; NC); (2) basal diet + 0.01% enramycin (positive control; PC); (3) basal diet + 3% non-fermented CC juice (NCC; CC juice 10%, water 90%); and (4) basal diet + 3% fermented CC juice (FCC; CC juice 10%, water 90%, Lactobacillus plantarum SK4719). Feed and water were provided ad libitum. Intriguingly, all treatments showed similar results in terms of broiler productivity and chicken meat quality. Considering organ characteristics, the FCC group showed a low spleen weight and lower (p < 0.05) blood levels of AST and total cholesterol (TCHO). Regarding intestinal characteristics, the CC feed additive (NCC and FCC) resulted in a heavier intestinal weight (p < 0.05) without affecting the length ratio of the villi or the crypt compared to the control (NC or PC). NCC and FCC lowered the growth of intestinal pathogens (p < 0.01). In summary, the addition of FCC can maintain poultry health by improving blood compositions and inhibiting the growth of intestinal pathogens, leading to a productivity comparable to that of poultry treated with growth-promoting antibiotics.

Potential Probiotic Acceptability of a Novel Strain of Paenibacillus konkukensis SK 3146 and Its Dietary Effects on Growth Performance, Intestinal Microbiota, and Meat Quality in Broilers.

This study evaluates the in vitro probiotic characteristics of P. konkukensis sp. nov. SK-3146, which was isolated from animal feed, and its dietary effects on growth performance, intestinal characteristics, intestinal microbiota, and meat quality in broilers. In vitro experiments revealed that P. konkukensis was non-hemolytic with variable antibiotic susceptibility, and acid as well as bile tolerance. To assess the effect of P. konkukensis on broilers, a total of four hundred eighty 1-day-old Ross 308 broiler chicks were allocated to 3 treatment groups with 4 replicates of 40 birds each; the negative control group was fed a basal diet without any feed additives (NC), the positive control group was fed a basal diet containing 0.01% enramycin (PC), and the experimental group was fed a basal diet containing P. konkukensis bacterial culture (PK) at 104 CFU/g of the diet based on bacterial count. The experiment lasted for 35 days. Results indicated that there were no significant differences in any growth performance parameters among the dietary treatments (p > 0.05). In addition, the inclusion of P. konkukensis in the broilers' diet did not affect meat cooking loss, color, and pH but increased the relative weight of breast meat (p < 0.05). The PK group showed heavier intestinal weight and shorter intestinal length than the NC group (p < 0.05). The ratio of the intestinal weight to length of jejunum was the highest in the PK group (p < 0.05). The PK group showed increased counts of Streptococcus thermophilus (p < 0.05) with no adverse effects of P. konkukensis on other intestinal microbiota in the jejunum. This study implies that P. konkukensis might have the potential to be applied as a probiotic feed additive in poultry.

Effect of Providing Environmental Enrichment into Aviary House on the Welfare of Laying Hens.

This study aimed to determine the effects of providing environmental enrichment materials­pumice stone and alfalfa hay­to laying hens in the aviary system. A total of 2196 40-week-old Hy-Line Brown laying hens were randomly allotted to three treatment groups: (1) no enrichment (control; CON), (2) enrichment with pumice stone (PS), and (3) enrichment with alfalfa hay (HAY). Each treatment comprised four replicates of 183 hens each, and four of the same materials were provided per replicate. The experiment lasted for 26 weeks. Feed and water were provided ad libitum. As a result, the PS and HAY groups demonstrated increased egg production (p < 0.001). The HAY group showed a reduced rate of mislaid eggs (p < 0.01) and produced low egg weight and pale-yellow yolk (p < 0.05). Both enrichment materials decreased blood creatinine (CRE) or lactate dehydrogenase (LDH) in the blood and resulted in a significantly lower corticosterone (CORT) level (p < 0.05). However, the feather condition scores for the laying hens were similar across all treatments (p > 0.05). In summary, although pumice stone and alfalfa hay are effective in alleviating stress and improving the production of laying hens, additional environmental improvement studies are needed to contribute to reducing pecking behaviors in poultry farming.

Feasibility of sodium long chain polyphosphate as a potential growth promoter in broilers.

The objective of this study was to evaluate in vitro antimicrobial and anti-biofilm activity of sodium long chain polyphosphate (SLCPP) and effect of dietary supplementation of SLCPP on growth performance, organ characteristics, blood metabolites, and intestinal microflora of broilers. Antimicrobial activities of SLCPP were observed against Escherichia coli O157:H7, Listeria monocytogenes, Salmonella enterica ser. Pullorum, Shigella sonnei, Klebsiella pneumonia, Pseudomonas aeruginosa in agar well diffusion assay. In addition, SLCPP demonstrated good anti-biofilm activity against K. pneumonia and P. aeruginosa. Furthermore, to investigate the dietary effect of SLCPP, a total of 480 1-day-old male Ross 308 broiler chicks were randomly allotted to three dietary treatment groups (4 replicates per group, 40 birds in each replicate): an antibiotic-free corn-soybean meal basal diet (NC); basal diet + enramycin 0.01% (PC); and basal diet + 0.1% SLCPP (SPP). The experiment lasted for 35 days. Results showed that birds fed with SLCPP had higher body weight (BW) and average daily gain (ADG), and lower feed conversion ratio (FCR) during the grower phase (days 7 to 21) (p < 0.05). Except for blood urea nitrogen, all other blood biochemical parameters remained unaffected by the dietary supplementation of SLCPP. Compared to the control group, lengths of the duodenum and ileum in the SPP group were significantly shorter (p < 0.05). Moreover, counts of lactic acid bacteria (LAB), total aerobes, and Streptococcus spp. in jejunum as well as LAB in cecum were increased in the SPP group than in the PC group (p < 0.05). These results suggest that dietary supplementation of SLCPP might promote the growth of broilers in their early growth phase.

Effect of Dietary Supplementation of Fermented Pine Needle Extract on Productive Performance, Egg Quality, and Serum Lipid Parameters in Laying Hens.

This study aimed to investigate the supplemental effects of fermented pine (Pinus densiflora) needle extract (FPNE) in laying hen diets on productive performance, egg quality, and serum lipid metabolites. A total of 108 40-week-old Hy-line brown laying hens were randomly assigned to one of the three dietary treatment groups: (1) basal diet + 0 mL FPNE/kg diet (CON), (2) basal diet + 2.5 mL FPNE/kg diet (T1), or (3) basal diet + 5 mL FPNE/kg diet (T2) for 6 weeks. Each group consisted of four replicates of nine hens each. Feed and water provided ad libitum. Results showed that dietary supplementation of FPNE increased egg production percentage (linear, p < 0.01 and quadratic, p < 0.05), egg mass (linear, p < 0.05), and feed intake (linear, p < 0.05) during the entire experimental period. In addition, dietary inclusion of FPNE significantly increased the eggshell color (linear, p < 0.01), egg yolk color (quadratic, p < 0.01), and eggshell breaking strength (linear, p < 0.05 and quadratic, p < 0.05) while the Haugh unit decreased (quadratic, p < 0.05). However, serum lipid profile did not differ among the dietary treatments (p > 0.05). Notably, antioxidant activity of egg yolk was improved by significantly decreasing the malondialdehyde content in egg yolks after 6 weeks of storage (linear, p < 0.001 and quadratic, p < 0.05). In summary, dietary inclusion of FPNE could improve laying performance and the antioxidant capacity of eggs.

Superiority of coarse eggshell as a calcium source over limestone, cockle shell, oyster shell, and fine eggshell in old laying hens.

Chicken eggshell (ES) waste is a rich source of calcium carbonate (CaCO3); however, the potential of ES as dietary calcium (Ca) in old laying hens has not been explored. This study compared the effects of feeding limestone, cockle shell, oyster shell, fine ES, and coarse ES as the sole Ca source on production performance, egg quality, blood biochemical constituents, and tibia characteristics in old laying hens. A total of 450 ISA-Brown laying hens at 73 wk of age with similar egg production rate (EPR) were randomly assigned to 5 treatment groups (90 hens/group, 9 hens/replicate) for 7 wk. Dietary treatment groups comprised a corn-soybean meal based diet containing different Ca sources: (i) limestone (LS; < 2 mm and 2-4 mm mixed in the ratio of 3:7) as control, (ii) cockle shell (CS; 1-4 mm), (iii) oyster shell (OS; 3-16 mm), (iv) ES fine particles (ESF; < 1 mm), and (v) ES coarse particles (ESC; 3-5 mm). Results indicated that dietary inclusion of coarse ES particles significantly increased average egg weight (P < 0.001) and daily egg mass (P < 0.05), and decreased feed conversion ratio (P < 0.001) as compared with the other treatments. However, no significant differences in EPR, feed intake, cracked egg proportion, and mortality were observed among the dietary treatments (P > 0.05). Notably, the use of ESF led to a lower proportion of cracked eggs than ESC (P < 0.05). ESC fed hens produced the heaviest eggs whereas CS fed hens produced the lightest (P < 0.001); the particle size of ES also affected the egg weight (P < 0.05). The eggs from OS and ESC fed hens showed a greater albumen height in comparison to eggs from CS group (P < 0.05); but no significant difference was observed among the LS, OS, ESF, and ESC groups (P > 0.05). The yolk color was darker in the eggs of group ESF as compared with other dietary groups (P < 0.01). However, no significant effects on Haugh units and shell properties were observed among the treatments (P > 0.05). The blood biochemistry results were not affected by the dietary Ca (P > 0.05) except for lower levels of high-density lipoprotein percentage (HDL %) in OS and ESC fed hens (P < 0.05). The tibia characteristics including weight, length, width, and breaking strength did not differ among the dietary groups (P > 0.05). However, the ESC and OS fed hens showed higher tibia bone mineral density (BMD) than the other groups (P < 0.001). In conclusion, coarse ES as a sole Ca source had beneficial effects on the production performance, egg quality, and tibia BMD in old laying hens.

Probiotic Potential of the Farmed Olive Flounder, Paralichthys olivaceus, Autochthonous Gut Microbiota.

In recent years, considerable and growing attention has been given to the application of host-associated microorganisms as a more suitable source of probiotics in aquaculture sector. Herein, we isolated and screened the olive flounder gut microbiota for beneficial bacterial strains that might serve as potential probiotics in a low fishmeal extruded aquafeed. Among the ten identified isolates, Bacillus amyloliquefaciens SK4079 and B. subtilis SK4082 were screened out based on their heat-resistant ability as well as enzymatic and non-hemolytic activities. Although both strains were well able to utilize carboxymethyl cellulose (CMC), xylan, and soybean meal (SBM) as a single carbon source in the minimal nutrient M9 medium, B. subtilis exhibited significantly higher cellulase, xylanase, and protease activities than B. amyloliquefaciens. The two selected strains were well able to degrade the undesirable anti-nutritional component of the SBM, which would limit its utilization as protein source in aquafeed industry. Significantly higher biofilm formation capacity and notably stronger adhesive interactions with the flounder's skin mucus were detected in B. subtilis than B. amyloliquefaciens. Immobilization of the spores from the selected strains, in a SBM complex carrier, remarkably enhances their thermal resistance at 120 °C for 5 min and different drying conditions. It was also interesting to learn that the B. subtilis spores could survive and remain viable after being sprayed onto extruded low-fish meal feed pellets for as long as 6 months. Overall, the findings of the present study could help the food/feed industries achieve their goal of developing cost-effective yet efficient products.

Effect of dietary supplementation of a phytogenic blend containing Schisandra chinensis, Pinus densiflora, and Allium tuberosum on productivity, egg quality, and health parameters in laying hens.

OBJECTIVE: This study was conducted to investigate the supplementary effect of a phytogenic blend (SPA: a mixture containing fermented Schisandra chinensis pomace, fermented Pinus densiflora needle extract, and Allium tuberosum powder in the ratio of 2:2:1) on egg production, egg quality, blood constituents, and visceral organs in laying hens. METHODS: A total of 135 Hy-line brown laying hens (48-wk-old) were randomly allocated to three dietary treatments with 5 replicates of 9 hens. The control group (CON) was fed a basal diet (no exogenous SPA addition) and the experimental groups were fed the basal diet containing SPA at the level of 0.1% and 0.3% for 6 weeks. RESULTS: The feed intake was significantly improved in SPA supplemented groups as compared with the control (p<0.05). However, egg production, daily egg mass, and feed conversion ratio were not different among the dietary treatments (p>0.05). For egg quality traits, only Haugh unit (HU) was significantly improved in SPA (0.3%) (p<0.05) as compared with other groups. However, HU was not affected during 4-wk of storage at 18°C among the dietary treatments (p>0.05). Furthermore, SPA supplementation did not affect the blood biochemical constituents except for the phosphate content, which was significantly higher in SPA groups than the CON group (p<0.05). There were no significant differences in visceral organ characteristics and immune indicators (immunoglobulin A [IgA], IgG, and IgM) in SPA or CON groups. CONCLUSION: This study suggested that the supplementation of SPA may have beneficial effects on feed intake and egg quality in laying hens.

Allium Flavonols: Health Benefits, Molecular Targets, and Bioavailability.

Allium species are revered worldwide as vegetables, condiments, and spices as well as the therapeutic agents in traditional medicine. The bioactive compounds in alliums mainly include organosulfur compounds, polyphenols, dietary fibers, and saponins. Flavonoids, particularly flavonols from alliums, have been demonstrated to have the antioxidant, anticancer, hypolipidemic, anti-diabetic, cardioprotective, neuroprotective, and antimicrobial activities. However, flavonols are mostly characterized from onions and have not been comprehensively reviewed across different species. This article therefore focuses on flavonol profiles from different Allium species, their health effects, underlying molecular mechanisms, and bioavailability. Intriguingly, the functional health effects of flavonols were mainly ascribed to their antioxidant and anti-inflammatory activities involving a cascade of multiple signaling pathways. Although the Allium-derived flavonols offer tremendous potential in preventing chronic disease risks, in-depth studies are needed to translate their clinical application.

Controlled Fermentation Using Autochthonous Lactobacillus plantarum Improves Antimicrobial Potential of Chinese Chives against Poultry Pathogens.

Chinese chives (CC) are rich in several antimicrobial constituents including organosulfur compounds, phenolics, and saponins, among others. Herein, we fermented CC juice using an autochthonous isolate, Lactobacillus plantarum having antimicrobial effects against poultry pathogens toward formulating an antimicrobial feed additive. Following 24 h of fermentation, the antimicrobial and antiviral activities of CC juice were significantly enhanced against poultry pathogens. However, the antioxidant activity of CC juice was significantly decreased following fermentation. Meanwhile, the compositional changes of CC juice following fermentation were also investigated. The total polyphenol, thiol, and allicin contents were significantly decreased in L. plantarum 24 h-fermented CC juice (LpCC) extract; however, total flavonoids increased significantly following fermentation. The untargeted metabolite profiling of nonfermented CC juice (NCC) and LpCC extracts was carried out using the ultra-high-performance liquid chromatography-linear trap quadrupole-orbitrap-tandem mass spectrometry (UHPLC-LTQ-Orbitrap-MS/MS) followed by multivariate analyses. The score plots of principal component analysis (PCA) and orthogonal partial least squares-discriminant analysis (OPLS-DA) based on UHPLC-LTQ-Orbitrap-MS/MS datasets displayed a clear segregation between the LpCC and NCC samples, which suggests their marked metabolomic disparity. Based on the multivariate analysis, we selected 17 significantly discriminant metabolites belonging to the different chemical classes including alkaloid, flavonols, saponins, fatty acids, amino acids, and organic acids. Notably, the flavonols including the glycosides of quercetin, kaempferol, and isorhamnetin as well as the saponins displayed significantly higher relative abundance in LpCC as compared with NCC. This study provides useful insights for the development of a fermented CC juice based antimicrobial feed additive to combat poultry infections.

Dietary effect of low fish meal aquafeed on gut microbiota in olive flounder (Paralichthys olivaceus) at different growth stages.

This study was conducted to investigate the long-term effect of a low fish meal (FM) diet comprising plant-based protein sources (PPS) on changes of gut microbial diversity in olive flounder (Paralichthys olivaceus) over the course of life. Two experimental diets were prepared to contain 74% FM (control) or 52% FM with 22% PPS (30% FM replacement, FM30). Fish were fed one of the two experimental diets for 8 months, and we collected the midgut contents to analyze the gut bacterial community by Illumina MiSeq based on the metagenomic sequences in the V3-V4 regions of 16S rRNA. We found that there were nine dominant phyla, which in turn presented Proteobacteria, Firmicutes, and Actinobacteria as the three major phyla in the gut microbiota of the flounder. At genus level, the dominant genera were Delftia, Prevotella, and Chthoniobacter at the juvenile stage (below 100 g/fish); Chthoniobacter, Bacillus, and Bradyrhizobium at the grower stage (400 g/fish); Chthoniobacter, Bacillus, and Delftia at the subadult stage (800 g/fish); and Lactobacillus and Prevotella at the adult stage (over 1,000 g/fish). The microbial diversity in olive flounders arched from the juvenile and subadult stage and reached a plateau thereafter. The fish fed the FM30 diet significantly had an increased abundance of Lactobacillus and Photobacterium and had less abundance of Prevotella and Paraprevotella than the control. However, the effect of dietary PPS was not significant on total microbial richness, indicating no negative effect as feed sources on the intestinal microbiota in olive flounder. These results indicate that the life stage of olive flounder is more important in modulating intestinal microbiota than is the diet. It could also be concluded that dietary PPS might be used as a potential fish meal alternative without any compromising effects on microbial diversity of olive flounder for long-term feeding.

The Genus Allium as Poultry Feed Additive: A Review.

The genus Allium, belonging to the family Amaryllidaceae has been known since ancient times for their therapeutic potentials. As the number of multi-drug resistant infections has increased due to in-feed antibiotic usage in poultry, the relevance of alliums as feed additives has been critically assessed. Garlic and the other Allium species, such as onions, leek, shallot, scallion, and chives, have been characterized to contain a plethora of bioactive compounds such as organosulfur compounds, polyphenols, saponins, fructans, and fructo-oligosaccharides. Consequently, alliums have been validated to confer antioxidant, antibacterial, antiviral, immunostimulatory, gut homeostasis, and lipid- as well as cholesterol-lowering properties in poultry. This review intends to summarize recent progress on the use of edible alliums as poultry feed additives, their beneficial effects, and the underlying mechanisms of their involvement in poultry nutrition. Perspectives for future research and limitations are also briefly discussed.

Effects of dietary multi-strain probiotics supplementation in a low fishmeal diet on growth performance, nutrient utilization, proximate composition, immune parameters, and gut microbiota of juvenile olive flounder (Paralichthys olivaceus).

A 12-week feeding trial was conducted to evaluate the effects of multi-strain probiotics (MSP) in a low fish meal (FM) diet on overall performance, gut microbiota, selected non-specific immune responses and antioxidant enzyme activities of olive flounder (Paralichthys olivaceus) juveniles. A total of 225 healthy olive flounders (initial mean body weight, 13.5 ±â€¯0.01 g) were randomly separated into 3 groups of 75 fish, each group having three replicates of 25 fish; first group was fed with a FM-based control diet (Con), 2nd group was fed with a low-FM diet containing a blend of plant and animal protein meals replacing 30% of the FM protein (FM30), and 3rd group was fed with the FM30 diet supplemented with 108-109 CFU kg-1 of the MSP (Pro). With the exception of lipid retention, which was significantly lower in fish fed the FM30 diet compared to the other two treatments, no other statistically significant differences were recorded with respect to any of the other growth and nutrient utilization parameters. Myeloperoxidase and lysozyme activities of fish fed the Pro diet were much higher and significantly different than those of fish fed the FM30 diet. Glutathione peroxidase activity was significantly higher in Pro- than in Con-fed fish, which, in turn, was significantly higher than FM30-fed fish. Expression of immune-related genes including IL-1ß, IL-6, and TNF-α was markedly upregulated in livers of the fish fed Pro diet compared to those fed the Con and FM30 diets. Furthermore, supplementation of MSP in FM30 diet enriched the Lactobacillus abundance in the fish gut as well as predictive gene functions in relation to lipid and carbohydrate metabolisms. These data suggested that the MSP could reduce the potential adverse effects of the low-FM diet and might be used as a healthy immunostimulant for olive flounder.

Autochthonous Bacillus licheniformis: Probiotic potential and survival ability in low-fishmeal extruded pellet aquafeed.

In recent years, Bacillus spp. have garnered attention as probiotic supplements in aquafeed owing to the production of heat stable and low pH resistant spores. Herein, we isolated and characterized an autochthonous Bacillus licheniformis KCCM 43270 from the intestine of olive flounder (Paralichthys olivaceus) for supplementation in low-fishmeal extruded aquafeeds. The KCCM 43270 was screened based on amylase, protease, cellulase, and lipase as well as non-hemolytic activities. The isolate was able to grow in carboxymethyl cellulose (CMC), xylan, and soybean meal (SBM) when used as a single carbon source in the minimal nutrient M9 medium. The KCCM 43270 spores displayed complete survival in acid (pH 2.5) and bile (0.3%, w/v) for 3 hr, strong biofilm formation, and nearly 50% adhesion with intestinal mucus. The spores of the isolate also showed significant survival ability at 80, 90, 100°C for 60, 30, and 1 min, respectively. In addition, the spores in a blend of SBM complex carrier showed significant heat stability at 120°C for 5 min and under different drying conditions. Furthermore, the spores also survived the extrusion process during low-fishmeal aquafeed manufacturing, implying the potential application of B. licheniformis KCCM 43270 in aquafeed industry.