Production of Spirulina platensis Using Inexpensive Local Resources in Palestine

Spirulina platensis is a special and unique cyanobacteria that is produced worldwide with a varied cost of cultivation media. In this study, five main experiments with different treatments were performed to evaluate the possibility of using cheap aquaculture water for Spirulina production, to test if solutions made by plant ash (PAS) could be used for Spirulina production, to determine if brackish water (BW) and mining water have a good impact on Spirulina production, to create a medium composed of cheap chemicals and fertilizers to be used for Spirulina cultivation, and to test if a mixture made from local components could be used to produce Spirulina. All experiments were performed via growth and dry weight measurements, including determination of chemical and physical characteristics of the samples with a comparison with Zarrouk medium (ZM) as a reference for each experiment, and all experiments were performed for 21 days to determine the best media type that lasts longer for commercial purposes. In all experiments, pH values were between 8 and 11, and EC was between 9.8 and 30 ms/cm, while temperature was at 30°C and 35°C, and light was at 1500 and 5000 Lux for 16 h light and 8 h dark. Spirulina can grow in (FW). It can also grow in FW diluted with BW. Also a 3% PAS can be used as a source to cultivate Spirulina at a very low price compared to ZM. The chemical fertilizer formula was one of the best types among all treatments with a good price. A mixture of these local resources could be a very good cheap alternative source. The main result that was obtained from all experiments in this study is the ability of Spirulina to grow within a wide range of chemical parameters at a lower price.

Comparison of the therapeutic effects of extracts from Spirulina platensis and amnion membrane on inflammation-associated corneal neovascularization

AIM: To compare the therapeutic effects of polysaccharide extract from Spirulina platensis (PSP) and extract from amnion membrane CAME) on alkali burn-induced corneal neovascularization (CorNV). METHODS: PSP and AME were Extracted from dry powder of Spirulina platensis and human aminion membrane respectively. Murine CorNV was induced by applying 1N sodiumhydroxide (NaOH) solution directly on the mice corneas. PSP and AME extracts were administered topically on the corneas 4 times daily for 7 days. The therapy effects of PSP and AME extracts were evaluated daily using slit-lamp. At the end of the therapy, corneas were harvested for H&E staining, masson trichrome staining, immunohistochemical study, and semi-quantification reverse transcriptive PCR (RT-PCR) was utilized for measurement of inflammation-related molecules. RESULTS: Topical application of PSP extract had significant therapeutic effects on CorNV that could be shown in various assays of the corneas. Compared with AME Extract, PSP extract treatment was more effective in suppressing CorNV in berms of vessel length and levels of cluster of differentiation 31 (CD31) proteins or the angiogenesis related genes like vascular endothelial growth factor (VEGF), matrix metalloproteinase-2 (MMP2) and matrix metalloproteinase-9 (MMP9). PSP also inhibited inflammation more markedly by more effectively inhibiting mononuclear and polymorphonuclear cells infiltration into the corneal stroma and reducing levels of stromal cell-derived factor-1 (SDF1), tumor necrosis factor-alpha (TNF alpha) and macrophage inflammatory protein-3 (MIP3a). In addition, corneas of PSP group had a more regular and compact architecture of collagen with thinner corneal thickness than in the AME group. ' CONCLUSION: Polysaccharide extract from Spirulina platensis inhibited alkali burn-induced inflammation and CorNV more effectively than AME Extract at the studied doses, thus may be used for the therapy of corneal diseases involving neovascularization and inflammation.

Protective effects of a polysaccharide from Spirulina platensis on dopaminergic neurons in an MPT-Pinduced Parkinson's disease model in C57BL/6J mice

The present study aimed to determine whether a polysaccharide obtained fromSpirulina platensis shows protective effects on dopaminergic neurons. A Parkinson’s disease model was established through the intraperitoneal injection of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyr-idine （MPTP） in C57BL/6J mice. Prior to the MPTP injection, some mice were pretreated with intraperitoneal injections of a polysaccharide derived fromSpirulina platensis once daily for 10 days. The results showed that the immunoreactive staining and mRNA expression of the dopa-mine transporter and tyrosine hydroxylase, the rate-limiting enzyme in dopamine synthesis, in the substantia nigra, were signiifcantly increased in mice pretreated with 800 mg/kg of the poly-saccharide compared with those in MPTP-treated mice. The activities of superoxide dismutase and glutathione peroxidase in the serum and midbrain were also increased signiifcantly in mice injected with MPTP after pretreatment with the polysaccharide fromSpirulina platensis. By con-trast, the activity of monoamine oxidase B in serum and midbrain maintained unchanged. These experimental ifndings indicate that the polysaccharide obtained fromSpirulina platensis plays a protective role against the MPTP-induced loss of dopaminergic neurons in C57BL/6J mice, and that the antioxidative properties of this polysaccharide likely underlie its neuroprotective effect.

Effect of Supplementation of Spirulina platensis to Enhance the Zinc Status in Plants of Amaranthus gangeticus, Phaseolus aureus and Tomato

Trace elements or micronutrients play a major role in the metabolism of both plants and humans. Zinc has a major specific role in metabolism when compared with other elements. The microbial biofertilizers are applied in the form of seaweed liquid extracts, microbial inoculants, biostimulators and biofortification agents. All these categories of microbial biofertilizers are involved in the enhancement of plant nutrient uptake. In the present study, Spirulina platensis is used as a biofortification agent to enhance zinc levels in cultivars of Amaranthus gangeticus, Phaseolus aureus and Tomato. Different experimental methods were followed including soaking seeds in different concentrations of Spirulina (5, 10, 15, 20, 25 and 30 g in 100 ml of water);soaking seeds in Spirulina hydrolysate at different time intervals (1, 2, 3, 4, 5 hrs and overnight);Spirulina in combination with biofertilizers, chemical fertilizer, organic fertilizer and vermicompost in various proportions (25:75;50:50;75:25) and foliar spray with different concentrations of Spirulina (25, 50, 75, and 100g in 5 litres of water) respectively. The zinc content of the yield of the cultivars was estimated and the study results indicated that there was a significant increase in zinc levels with p-value 0.015, 0.003 and 0.035 for Amaranthus gangeticus, Phaseolus aureus and Tomato respectively when compared with the control and between the set-ups, with biofortification of Spirulina platensis. The obtained results emphasize the application of Spirulina platensis to enhance the mineral nutrient in plants which are non-polluting, inexpensive, utilizable renewable resource to maintain the soil fertility.

The Stabilization Effect of Glutaraldehyde on the Spirulina platensis Phycobilisomes

The spectral properties of the glutaraldehyde-treated phycobilisomes were studied. The results showed that glutaraldehyde was effective in preventing phycobilisomes from dilution- induced dissociation and preserving the intra-phycobilisomes energy transfer.

Isolation and Characterization of C-phycocyanin from Spirulina Platensis

ＩｎｔｒｏｄｕｃｔｉｏｎＳｐｉｒｕｌｉｎａ，ａｋｉｎｄｏｆｆｉｌｉｆｏｒｍ，ｍｕｌｔｉｃｅｌｕｌａｒａｌｇａｅｔｈａｔｉｓｒｉｃｈｉｎｐｒｏｔｅｉｎ，ｆａｔ，ｖｉｔａｍｉｎ，ｍｉｎｅｒａｌ，ｃｈｌｏｒｏｐｈｙｌ，β－ｃａｒｏｔｅｎｅａｎｄｐｏｌｙｓａ...

The prevention role of Spirulina platensis(Arthrospira platensis)on intestinal health

Spirulina is a popular herbal food that has a preventive effect on health.In the study,it aimed to evaluate the effects of different doses of Spirulina platensis(Arthrospira platensis)on the morphological properties of the duodenum of rats,and to assign the effect of Spirulina on the expression of proliferating cell nuclear antigen(PCNA)in the rat duodenum,and thereby to observe the effects on intestinal health.30 male Wistar albino rats were divided into 3 groups for 45 days.The first group was received the basal diet;the second group was given 500 mg/kg Spirulina daily by oral gavage;the third group was given 1000 mg/kg Spirulina daily.The duodenum segments were taken at the end of the trial and processed for histological assay.Although the total mucosa,villus height and villus/cript ratio were found higher in high dose Spirulina,the lower cript depth was detected in same group compared to control and low dose group.A significant increase was observed at high dose compared to the control and low dose group in terms of PCNA expression intensity and proliferation index.Findings suggest that high dose of Spirulina may support the duodenal growth,and thereby intestinal health.

Structural characterization of oligosaccharide from Spirulina platensis and its effect on the faecal microbiota in vitro

In the present study,an oligosaccharide SPO-1 from Spirulina platensis was prepared by glycosidase from a marine bacterium.The prebiotic activity of SPO-1 on the growth of Lactobacillus paracasei and Bifidobacterium animalis,and its effect on human gut microbiota were examined in vitro.The molecular weight of the tetrasaccharide SPO-1 was 650.2 Da,and it was mainly composed of glucose with α-type glycosidic linkages.The prebiotic activity score of SPO-1 was the highest for the growth of probiotic strains L.paracasei and B.animalis.Furthermore,as fermentation proceeded,SPO-1 was gradually degraded and utilized by intestinal bacteria.The results showed that after treatment with SPO-1,carbohydrate consumption and short-chain fatty acids levels were increased,especially those of i-butyric and i-valeric acids.Moreover,SPO-1 significantly promoted the abundance,diversity and composition of gut microbiota,especially stimulating the growth of Bacteroides,Escherichia-Shigella and Megamonas.In addition,the change in intestinal microbiota function predicted by phylogenetic investigation of communities by reconstruction of unobserved states(PICRUSt)after treatment with SPO-1 is mainly related to the terms“carbohydrate metabolism”and“amino acid metabolism”.These results suggest that SPO-1 is a potential oligosaccharide in regulation of intestinal microbiota.

Physicochemical and Thermal Characterization of the Spirulina platensis

Cyanobacterium Spirulina platensis is commercially produced as a nutrient source for food,animal feed and pharmaceutical industries,and is also explored in other applications in areas such as material sciences,materials engineering and for the production of biofuels and biochemicals.Due to the increasing interest in the use of this microalga,a complete characterization was intended,as to provide data to the insufficient literature.In this work,various techniques were used for thermal(thermogravimetric analysis(TGA)/derivative thermogravimetry(DTG),differential scanning calorimetry(DSC)),structural(scanning electron microscopy(SEM)and wide-angle X-ray diffraction(XRD))and chemical(atomic absorption spectroscopy(AAS),attenuated total reflection Fourier transform infrared spectroscopy(ATR-FTIR),energy dispersive X-ray spectroscopy(EDX))characterization of Spirulina cultivated in Brazil.Results have shown that in addition to the high quantity of protein(over 50%),Brazilian Spirulina is a source for carbohydrates(33%)and also has good thermal stability up to 200°C.The pigment protein Phycocyanin could be identified by FTIR and ultraviolet-visible(UV-vis)spectroscopy.Results show favorable properties of Spirulina as a source for new materials and biomass.

Hematobiochemical and histopathological alterations in Nile Tilapia(Oreochromis niloticus)exposed to ethidium bromide:The protective role of Spirulina platensis

Ethidium bromide(EtBr)is one of the contaminants recorded in aquatic environments whose effects have been investigated;however,there is still limited knowledge about its remediation.This study examined the potential protective effects of Spirulina platensis(SP)against the effects of EtBr toxicity in the Nile tilapia(Oreochromis niloticus)fry.Fry were divided to five groups,viz.,a control and four treatment groups of low-dose EtBr(10μg/L),low-dose EtBr with SP(10μg/L EtBr+200 mg/L SP),high-dose EtBr(100μg/L),and high-dose EtBr with SP(100μg/L EtBr+200 mg/L SP);the exposure period was 2 weeks.Low and high doses of EtBr induced alterations in some hematological,biochemical,and histopathological parameters.Necrotic hepatocytes,degenerated area,vacuolated hepatocytes,pyknotic nuclei,constricted and dilated blood sinusoids,and infiltration of inflammatory cells were observed.Lipid peroxidation concentration was not significantly different in groups exposed to low doses of EtBr and EtBr with SP,but it was increased in groups exposed to high doses of EtBr and EtBr with SP,compared with the control group.After feeding with SP,most histological and histochemical parameters restored to normal values.Therefore,SP may possess the ability to preserve the structural integrity of the hepatic and renal membranes.

Selenium-induced Changes in Activities of Antioxidant Enzymes and Content of Photosynthetic Pigments in Spirulina platensis

Spirulina platensis exposed to various selenium （Se） concentrations （0, 10, 20, 40, 80, 150, 175, 200, 250 mg/L） accumulated high amounts of Se in a dose- and time-dependent manner. Under low Se concentrations （〈150 mg/L）, Se induced increases in biomass concentration, content of photosynthetic pigments, and activities of glutathione peroxidase （GPX）, superoxide dismutase （SOD）, catalase （CAT） and Gua-dep peroxidases （POD）, which indicates that antioxidant enzymes play an important role in protecting cells from Se stress. Higher Se concentrations （≥175 mg/L） led to higher Se accumulation and increases in activities of GPX, SOD, CAT and POD, but also induced lipid peroxidation （LPO） coupled with potassium leakage and decreases in biomass concentration and contents of photosynthetic pigment. The results indicate that increases in activities of the antioxidant enzymes were not sufficient to protect cell membranes against Se stress. Time-dependent variations in the activities of antioxidant enzymes, contents of chlorophyll a and carotenoid and the LPO level were also investigated under representative Se concentrations of 40 and 200mg/L. Opposite variation trends between SOD-CAT activities, and GPX-POD-APX activities were observed during the growth cycles. The results showed that the prevention of damage to cell membranes of S. platensis cells could be achieved by cooperative effects of SOD-CAT and GPX-POD-APX enzymes. This study concludes that S. platensis possessed tolerance to Se and could protect itself from phytotoxicity induced by Se by altering various metabolic processes.

Spirulina platensis aqueous extracts ameliorate colonic mucosal damage and modulate gut microbiota disorder in mice with ulcerative colitis by inhibiting inflammation and oxidative stress

Ulcerative colitis(UC)is a chronic and recurrent inflammatory bowel disease(IBD)that has become a major gastroenterologic problem during recent decades.Numerous complicating factors are involved in UC development such as oxidative stress,inflammation,and microbiota disorder.These factors exacerbate damage to the intestinal mucosal barrier.Spirulina platensis is a commercial alga with various biological activity that is widely used as a functional ingredient in food and beverage products.However,there have been few studies on the treatment of UC using S.platensis aqueous extracts(SP),and the underlying mechanism of action of SP against UC has not yet been elucidated.Herein,we aimed to investigate the modulatory effect of SP on microbiota disorders in UC mice and clarify the underlying mechanisms by which SP alleviates damage to the intestinal mucosal barrier.Dextran sulfate sodium(DSS)was used to establish a normal human colonic epithelial cell(NCM460)injury model and UC animal model.The mitochondrial membrane potential assay 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide(MTT)and staining with Annexin V-fluorescein isothiocyanate(FITC)/propidium iodide(PI)and Hoechst 33258 were carried out to determine the effects of SP on the NCM460 cell injury model.Moreover,hematoxylin and eosin(H&E)staining,transmission electron microscopy(TEM),enzyme-linked immunosorbent assay(ELISA),quantitative real-time polymerase chain reaction(qPCR),western blot,and 16S ribosomal DNA(rDNA)sequencing were used to explore the effects and underlying mechanisms of action of SP on UC in C57BL/6 mice.In vitro studies showed that SP alleviated DSS-induced NCM460 cell injury.SP also significantly reduced the excessive generation of intracellular reactive oxygen species(ROS)and prevented mitochondrial membrane potential reduction after DSS challenge.In vivo studies indicated that SP administration could alleviate the severity of DSS-induced colonic mucosal damage compared with the control group.Inhibition of inflammation and oxidative stress was associated with increases in the activity of antioxidant enzymes and the expression of tight junction proteins(TJs)post-SP treatment.SP improved gut microbiota disorder mainly by increasing antioxidant enzyme activity and the expression of TJs in the colon.Our findings demonstrate that the protective effect of SP against UC is based on its inhibition of pro-inflammatory cytokine overproduction,inhibition of DSS-induced ROS production,and enhanced expression of antioxidant enzymes and TJs in the colonic mucosal barrier.

Arsenic methylation by an arsenite S-adenosylmethionine methyltransferase from Spirulina platensis

Arsenic-contaminated water is a serious hazard for human health. Plankton plays a critical role in the fate and toxicity of arsenic in water by accumulation and biotransformation.Spirulina platensis（S. platensis）, a typical plankton, is often used as a supplement or feed for pharmacy and aquiculture, and may introduce arsenic into the food chain, resulting in a risk to human health. However, there are few studies about how S. platensis biotransforms arsenic. In this study, we investigated arsenic biotransformation by S. platensis. When exposed to arsenite（As（Ⅲ））, S. platensis accumulated arsenic up to 4.1 mg/kg dry weight.After exposure to As（Ⅲ）, arsenate（As（Ⅴ）） was the predominant species making up 64% to86% of the total arsenic. Monomethylarsenate（MMA（Ⅴ）） and dimethylarsenate（DMA（Ⅴ））were also detected. An arsenite S-adenosylmethionine methyltransferase from S. platensis（Sp Ars M） was identified and characterized. Sp Ars M showed low identity with other reported Ars M enzymes. The Escherichia coli AW3110 bearing Spars M gene resulted in As（Ⅲ） methylation and conferring resistance to As（Ⅲ）. The in vitro assay showed that Sp Ars M exhibited As（Ⅲ） methylation activity. DMA（Ⅴ） and a small amount of MMA（Ⅴ） were detected in the reaction system within 0.5 hr. A truncated Sp Ars M derivative lacking the last 34 residues still had the ability to methylate As（Ⅲ）. The three single mutants of Sp Ars M（C59S, C186 S, and C238S） abolished the capability of As（Ⅲ） methylation, suggesting the three cysteine residues are involved in catalysis. We propose that Sp Ars M is responsible for As methylation and detoxification of As（Ⅲ） and may contribute to As biogeochemistry.

Effect of CeCl_3 on the Photosynthetic Characteristics of Spirulina Platensis

In this paper we report that CeCl3 could accelerate the synthesis of chlorophyll a and protein in spirulina platensis. and enhance the activity of oxygen evolution. We also found that the accelerations of CeCl3 in spirulina platensis culture solution of low NaHCO3 were more than in culture solution of high NaHCO3, therefore it was related to the concentration of NaHCO3 The absorption spectrum and low temperature (77 K) fluorescence emission spectrum of spirulina platensis with CeCl3 treated were significantly different from their controls,i.e.its absorption peak of phycocyanin at 620 nm was higher than that of the control.its absorption peak at the long wave region migrated from 676 to 680 nm, and its fluorecence emission ratios (730 nm/685～695 nm) of photosystem I(PSI) to photosystem Ⅱ(PS Ⅱ) were also higher than those of the control; the pitch of the spiral algal body became shorter, and induced structural changes too.

Effect of Spirulina Platensis Polysaccharide on Hematopoietic Recovery and Related Cytokines in Mice with Transplanted Tumor Treated by Chemotherapy

Objective:To evaluate the effect of Spirulina platensis polysaccharide (SPP) on hematopoietic recovery and related cytokines in mice with transplanted tumor after chemotherapy. Methods:Mouse model of transplanted solid tumor was established and treated with chemotherapy. Peripheral blood cells, bone marrow nucleated cells, and colony forming unit spleen (CFU S) were counted; the content of DNA in bone marrow was inspected by ultraviolet spectrophotometer; serum content of cytokines, interleukin (IL)1, IL 3, granulocyte macrophage colony stimulating factor (GM CSF) and tumor necrosis factor α (TNF α) were determined by double antibody sandwich ELISA.Results:Cyclophosphamide (CTX) could induce evident myelosuppression, manifested as decrease of peripheral blood cells, bone marrow nucleated cell and DNA, and the CFU S number. SPP could significantly ameliorate the myelosuppression induced by CTX without reducing anti tumor effect of CTX. In addition, it could also increase the contents of IL 1, IL 3, GM CSF, TNF α in serum. Conclusion:SPP can probably accelerate the hematopoietic recovery in mice after chemotherapy through promoting endogenous secretion of cytokines.

Isolation and structural elucidation of a polysaccharide with radiation-protective activity from Spirulina platensis

The structure of a radiation-protective sulfated polysaccharide, SP-APS, isolated from Spirulina platensis, was investigated by methylation analysis, partial acetolysis and H-1-NMR spectroscopy. The repeating-unit of SP-APS as branched structures was elucidated.

Purification and anti-inflammatory effect of selenium-containing protein fraction from selenium-enriched Spirulina platensis

Spirulina platensis is an excellent carrier of Se,and thus widely used in medical fields and as well as in the food industry.However,there is little information about the characteristics and bioactivity of selenium-containing S.platensis proteins(Se-SP).In this study,Se-SP with different molecular weights were isolated from selenium-enriched S.platensis,and the bioactivities(such as antioxidant and anti-inflammatory activities)of Se-SP were investigated.Se-SP3(with a molecular weight range of 20-48 kDa)showed better free radical scavenging ability(ABTS)than the other Se-SPs.In addition,Se-SP3 suppressed inflammatory cytokines,in which decreased by 74% interleukin 6(IL-6),42.28% tumor necrosis factor-α(TNF-α),69.07% content of malondialdehyde(MDA),40.45% interleukin-1β(IL-1β)relative to the LPS group.Moreover,Se-SP3 decreased the nitric oxide(NO)production by 64.84% compared with the LPS group and increased the activities of superoxide dismutase(SOD)and glutathione peroxidase(GSH-Px),indicating that Se-SP3 has excellent antioxidant and anti-inflammatory activities,and could be used as a functional food ingredient or natural medicine.

Antibacterial and anticancerous biocompatible silver nanoparticles synthesised from the cold-tolerant strain of Spirulina platensis

Objective:To synthesize silver nanoparticles from the biomass of cold tolerant strain of Spirulina platensis and evalute the synthesized nanoparticles against antibacterial and anticancer activity.Methods:Silver nanoparticles were synthesized by the algal culture and characterized by UV-vis spectroscopy,Fourier transform infrared spectroscopy,field emission scanning electron microscopy and X ray diffraction studies.Antibacterial activity has been studied with free nanoparticles adopting agar diffusion assay,biofilm inhibition assay and nanoparticles fabricated wound dressing against representative Gram-negative organism Pseudomonas aeruginosa and Gram-positive organism Staphylococcus aureus respectively.The in vitro anticancer activity of silver nanoparticles were screened against human Hep2 cell lines by means of MTT assay.Results:Reduction of silver ions by the algal culture was observed during 72 h of incubation and the synthesized nanoparticles were further characterized.Antibacterial study reveals both the strains were susceptible to free nanoparticles and fabricated wound dressing treatment.The in vitro anticancer activity of silver nanoparticles were screened against human Hep 2 cell lines by means of MTT assay which reveals that cell viability has been reduced as dose dependent manner.Conclusions:The observed results imply that silver nanoparticles synthesized from Spirulina platensis cold tolerant strain can be used as potential antibacterial and anticancerous agent.

Effects of the Partial Replacement of Soybean Meal by Insect or Algae Meal in Chicken Diets with Graded Amino Acid Supply on Parameters of Gut Microbiology and Dietary Protein Quality

Insects or algae are considered future solutions to substitute imported feed proteins like soybean meal (SBM) in animal nutrition. The objective of this research, as part of the multidisciplinary project “Sustainability transitions”, focused on replacing 50% SBM by partly defatted larvae meal from black soldier flies (Hermetia illucens) or the blue-green algae Spirulina platensis in meat type chicken diets. The current study aimed to evaluate the effects on body composition, protein quality, apparent precaecal digestibility (apcD) and parameters of intestinal microbiota. In total, 288 one-day-old male growing chickens (Ross 308) from a commercial hatchery were randomly allotted to 48 pens (6 birds per pen) across five diets, which were fed at a free choice level. The control diet utilized SBM, wheat and corn as main ingredients, and experimental diets replaced 50% of SBM by the alternative proteins under study. Amino acid (AA) supplementation of the final diets was conducted both at a basic level (diets HM, SM;Lys and Met added equal to the control diet) and an extended level of AA fortification (diets HM+, SM+;extended supplementation of Lys, Met, Thr, Arg). After finishing the growth study, 4 individual birds per diet were slaughtered for whole body analyses to derive nutrient utilization and dietary protein quality parameters. Additionally, pooled chyme samples from 16 birds per diet (control, HM and SM) were analyzed to assess apcD and microbial parameters. Diets HM and SM with a basic level of AA fortification led to significant depressions in nutrient deposition and dietary protein quality. However, HM+ and SM+ diets with an extended level of AA supplementation led to significantly improved responses;however these were still generally not on par with the control diet. The HM+ diet provided superior CP deposition and dietary protein quality, as compared to the SM+ diet, but equal to the control diet. Results of gut microbiology yielded no significant effects due to feeding the alternative protein sources under study.