Effect of baker's yeast fermented moist feed on the growth and bone mineralization in broiler.

Objective: The effect of feeding yeast-fermented feed in various forms on broiler growth performance and bone mineralization was studied. Materials and Methods: Initially, a corn-soy-based diet was formulated and fermented in anaerobic conditions at 28°C in laboratory space for 48 h with yeast (2.0%) and moisture (50%). Afterward, the 150 newly hatched Arbeor Acres commercial broiler chicks were divided into 5 dietary groups (30 chicks, 6 cages, and 5 birds per cage). Each group received one of the following formulated and fermented diets: dry feed (DF), moist feed (MF), yeast-added dry feed (Y-DF), yeast-added moist feed (Y-MF), or yeast-fermented moist feed (YF-MF). Water and feed were supplied ad libitum. Six birds per group were slaughtered at age 37 for the determination of carcass traits and tibia ash. Results: Fermentation improved crude protein from 20.7% to 22.8% but declined crude fiber from 7.9% to 6.3% in the YF-MF group compared to the DF group. High body weight gain was recorded in 771, 830, and 992 gm in the MF, Y-MF, and YF-MF groups, respectively, compared to the DF (762 gm) group (p < 0.01). The feed conversion ratio was better in the Y-MF (1.57) and YF-MF (1.57) groups than in the DF (1.75) group. Feeding a fermented, moist diet resulted in improved carcass yield (69%) in the YF-MF group. Bone mineralization expressed a better tibia ash percentage (35% from 30%) in the YF-MF group compared to the DF group. Conclusion: Therefore, YF-MF enhanced the quality of feed and improved growth, carcass weight, and bone mineralization in broiler.

Effect of dietary supplementation of plantain herb, lemongrass and their combination on milk yield, immunity, liver enzymes, serum, and milk mineral status in dairy cows.

Objective: This research aimed to assess the effects of dried plantain herb, lemongrass, and their combination on milk yield, immunological, liver enzymatic, serum, and milk mineral status in dairy cows. Materials and Methods: Twenty cows were arbitrarily assigned to 4 diets. Cows were given a basal ration considered as control diet (CL-D) having 14.93% crude protein (CP)and 10.96 MJ ME per kg dry matter (DM). Each cow was given 100 gm plantain, 100 gm lemongrass, and 50 gm plantain + 50 gm lemongrass with CL-D and taken as plantain diet (PT-D), lemongrass diet (LG-D), and plantain-lemongrass diet (PL-D), daily for 63 days, respectively. Blood and milk samples were taken four times at an interval of 14 days. Data were analyzed using a two-way repeated measures analysis of covariance. Results: Better DM consumption and milk yield were observed in the PT-D and LG-D compared to the CL-D (p ≤ 0.05). LG-D improved the milk's total solids, protein, and fat compared to CL-D (p < 0.05). Substantially, herbal groups improved serum albumin and reduced globulin concentrations compared to CL-D. LG-D had the highest serum immunoglobulin G, while herbal groups effectively reduced the liver enzymes compared to CL-D. Herbal groups did not affect serum and milk's calcium and phosphorus concentrations, while LG-D and PL-D substantially improved serum and milk zinc concentrations. Conclusions: Both plantain and lemongrass improved dairy cows' DM consumption and milk yield. Plantain and/or lemongrass enhanced the immune system and liver health, but not serum and milk calcium and phosphorus level. Lemongrass and a combination of plantain and lemongrass increased the serum and milk zinc concentrations.

Combined impact of lemongrass and spearmint herbs on performance, serum metabolites, liver enzymes, and meat quality of broiler.

Objectives: This study aimed to assess the influence of feeding fresh lemongrass (Cymbopogon citratus) or spearmint (Mentha spicata) and their combination on performance, serum metabolites, liver enzymes, and meat quality in broilers. Materials and Methods: A total of 168 day-old Indian River chicks were arbitrarily offered four experimental rations: (i) control ration (CT-R): corn-soya-based ration, (ii) lemongrass ration (LG-R): CT-R + 1.0% DM of lemongrass; (iii) spearmint ration (SM-R): CT-R + 1.0% DM of spearmint; and (iv) lemongrass-spearmint ration (LS-R): CT-R + 0.5% DM from both lemongrass and spearmint. Each ration was given to 42 birds for a duration of 35 days, with 3 replications and 14 birds each. Results: Elevated body weight gain was observed in LG-R (1,502 gm), LS-R (1,492 gm), and SM-R (1,474 gm) compared to CT-R (1,451 gm) (p = 0.078). Herbal rations successfully reduced almost 3%-5% of serum and meat total cholesterol concentrations compared to CT-R. Compared to CT-R, the highest zinc and iron concentrations of serum and meat were measured in LG-R and SM-R, respectively, while both minerals of serum and meat were observed to be better in LS-R (p < 0.05). Herbal rations significantly improved serum liver enzyme activity and ameliorated the red color of breast and thigh meat but failed to improve the lightness and yellowness of both types of meat compared to CT-R. Conclusions: LG-R, SM-R, and LS-R improved bird performance, liver health, and meat color, and lowered serum and meat cholesterol levels. But among them, LS-R efficaciously increased the serum and meat zinc and iron concentrations.

A Comprehensive Analysis into the Therapeutic Application of Natural Products as SIRT6 Modulators in Alzheimer's Disease, Aging, Cancer, Inflammation, and Diabetes.

Natural products have long been used as drugs to treat a wide array of human diseases. The lead compounds discovered from natural sources are used as novel templates for developing more potent and safer drugs. Natural products produce biological activity by binding with biological macromolecules, since natural products complement the protein-binding sites and natural product-protein interactions are already optimized in nature. Sirtuin 6 (SIRT6) is an NAD+ dependent histone deacetylase enzyme and a unique Sirtuin family member. It plays a crucial role in different molecular pathways linked to DNA repair, tumorigenesis, glycolysis, gluconeogenesis, neurodegeneration, cardiac hypertrophic responses, etc. Thus, it has emerged as an exciting target of several diseases such as cancer, neurodegenerative diseases, aging, diabetes, metabolic disorder, and heart disease. Recent studies have shown that natural compounds can act as modulators of SIRT6. In the current review, a list of natural products, their sources, and their mechanisms of SIRT6 activity modulation has been compiled. The potential application of these naturally occurring SIRT6 modulators in the amelioration of major human diseases such as Alzheimer's disease, aging, diabetes, inflammation, and cancer has also been delineated. Natural products such as isoquercetin, luteolin, and cyanidin act as SIRT6 activators, whereas vitexin, catechin, scutellarin, fucoidan, etc. work as SIRT6 inhibitors. It is noteworthy to mention that quercetin acts as both SIRT6 activator and inhibitor depending on its concentration used. Although none of them were found as highly selective and potent modulators of SIRT6, they could serve as the starting point for developing selective and highly potent scaffolds for SIRT6.

Phosphorus Availability in Laying Hens Given Non-phytate Phosphorus Deficient Diets Containing Buckwheat.

To date, the effect of buckwheat phytase on laying hens has not yet been investigated. Thus, this study was conducted to compare the production performance, egg quality, and phosphorus (P) balance of laying hens given nonphytate P deficient diets supplemented with non-germinated buckwheat (BU) or germinated buckwheat (GBU). Experimental diets (17.8% CP, 2,988 kcal/kg ME) consisted of two control diets, the positive control (PC), satisfying all nutrient requirements and negative control (NC) containing 0.16% less non-phytate P than that in the PC diet, and six experimental diets (containing 10%, 15%, 20% BU or GBU), prepared by replacing maize with BU or GBU, along with the raw materials of NC diet. Fifty-six laying hens (46 week of age) were allocated to eight dietary groups (seven hens each) and experimental diets were given for a period of six weeks (week one was employed for acclimatization, and the subsequent five weeks for data collection). Deteriorated production performance (hen-day egg production, feed intake, egg weight and egg mass) and eggshell quality (shell breaking strength, shell weight and shell thickness) in laying hens given a non-phytate P deficient NC diet was restored by the addition of at least 15% BU and 10% GBU to the NC diet. Total P retention significantly increased in 20% BU, 15% GBU and 20% GBU groups as excretion decreased considerably in these groups than the NC group. Considering the hen-day egg production as an economically important parameter, we found that a 340 phytase unit (PU)/kg diet of buckwheat phytase was equivalent to 0.10% non-phytate P in laying hens. These results suggested that the addition of buckwheat in non-phytate P deficient diets can alleviate the deficiency and improve P availability in laying hens.

Growth Performance, Bone Quality, and Phosphorus Availability in Broilers Given Phosphorus-Deficient Diets Containing Buckwheat (Fagopyrum esculentum).

To determine whether buckwheat phytase can be used as an alternative phytase source, growth performance, bone quality and P retention were measured in broilers given non-phytate P-deficient diets. Non-germinated (BU) and germinated (GBU) buckwheat were used: they were ground and sieved to remove hulls before use. A total of 120 male broiler chicks (8 d of age) were divided into 8 groups (15 birds each) and given one of the following 8 diets until 42 d of age: positive control (PC) diet satisfying recommended level of all nutrients, negative control (NC) diet formulated to contain 0.16% lower non-phytate P than PC diet, and six other diets, formulated by replacing maize in NC diet with BU or GBU at 10%, 15% and 20% concentrations. Starter diets contained 23.5% crude protein (CP) and 3,200 kcal of ME/kg, and were used for 8-21 d of age. Then, grower diets with 20.5% CP and 3,250 kcal of ME/kg, and were provided for 22-42 d of age. Compared with the PC group, NC group showed impaired growth performance (BW gain, FI, and FCR), and bone quality (dry weight, breaking strength and contents of ash and P in tibia). However, in most cases, these impairments were ameliorated dose-dependently by the addition of BU and GBU in diets, and the restoration magnitude was greater in GBU than in BU treatment. Total P excretion decreased in NC group and further decreased dose-dependently with increasing levels of BU and GBU. Except for the values in PC group, total P retention increased as the total P excretion decreased. In conclusion, dietary BU and GBU restored the growth performance and bone quality impaired by the P deficiency, and improved P retention in broilers, which suggested that buckwheat, especially when germinated, can be used as an alternative phytase source in broiler diets.

Phytase Activity in the Digesta from Different Parts of the Digestive Tract and Ileal Digestibility of Nutrients in Broilers Fed with Buckwheat Diets.

In the present study, the effects of dietary buckwheat on phytase activity in the digesta from different parts of the digestive tract, and ileal digestibility of nutrients were determined in broilers fed with buckwheat diets. Eighty male broilers (29-d-old) were divided into four groups (20 birds each), and were fed one of the following diets until they were 36-d-old: positive control (PC) diet formulated based on the NRC recommendations, negative control (NC) diet containing 0.15% lower non-phytate phosphorus (P) than that in the PC diet, and two other diets formulated by replacing corn in NC diet with either 20% non-germinated (BU) or germinated (GBU) buckwheat. At the age of 36 d, broilers were sacrificed to collect digesta from the crop, gizzard, duodenum, jejunum, ileum, and cecum. The activity of phytase was low in the PC and NC diets, which increased in the BU diet and increased further in the GBU diet. A similar trend was observed in the crop digesta; however, the phytase activity in the crop digesta of BU and GBU diets was marginally lower when compared with that in each diet. These values decreased sharply when the digesta moved to the gizzard, and then decreased gradually. The ileal digesta exhibited significantly low activity with negligible effect of dietary treatment. The result of two-way analysis of variance with germination and digestive tract parts as main factors showed that the effect of digestive tract parts and interaction between factors was significant on the phytase activity in digesta. The dietary BU and GBU did not affect the ileal crude protein digestibility; however, it increased the ileal phytate P digestibility. These results suggest that in broilers, the crop might be the primary site of phytate degradation by buckwheat phytase, and the buckwheat might have negligible adverse effect on ileal digestibility of nutrients.

Mycobacterial stress regulation: The Dps "twin sister" defense mechanism and structure-function relationship.

In this work, we have tried to emphasize the connection between mycobacterial growth and regulation of gene expression. Utilization of multiple carbon sources and diauxic growth helps bacteria to regulate gene expression at an optimum level so that the inhospitable conditions encountered during nutrient depletion can be circumvented. These aspects will be discussed with respect to mycobacterial growth in subsequent sections. Identification and characterization of genes induced under such conditions is helpful to understand the physiology of the bacterium. Although it is necessary to compare the total expression profile of proteins as they transit from vegetative growth to stationary phase, at times a lot of insights can be deciphered from the expression pattern of one or two proteins. We have compared the protein expression and sigma factor selectivity of two such proteins in M. smegmatis to understand the differential regulation of genes playing diverse function in the same species. Some newer insights on the structure and function of one of the Dps proteins are also explained.

The mycobacterial MsDps2 protein is a nucleoid-forming DNA binding protein regulated by sigma factors sigma and sigma.

The Dps (DNA-binding protein from starved cells) proteins from Mycobacterium smegmatis MsDps1 and MsDps2 are both DNA-binding proteins with some differences. While MsDps1 has two oligomeric states, with one of them responsible for DNA binding, MsDps2 has only one DNA-binding oligomeric state. Both the proteins however, show iron-binding activity. The MsDps1 protein has been shown previously to be induced under conditions of starvation and osmotic stress and is regulated by the extra cellular sigma factors sigma(H) and sigma(F). We show here, that the second Dps homologue in M. smegmatis, namely MsDps2, is purified in a DNA-bound form and exhibits nucleoid-like structures under the atomic force microscope. It appears that the N-terminal sequence of Dps2 plays a role in nucleoid formation. MsDps2, unlike MsDps1, does not show elevated expression in nutritionally starved or stationary phase conditions; rather its promoter is recognized by RNA polymerase containing sigma(A) or sigma(B), under in vitro conditions. We propose that due to the nucleoid-condensing ability, the expression of MsDps2 is tightly regulated inside the cells.

Molecular mechanism of in vitro oligomerization of Dps from Mycobacterium smegmatis: mutations of the residues identified by "interface cluster" analysis.

The irreversible dodecamerization of native Dps trimers from Mycobacterium smegmatis, in vitro, is known to be directly associated with the bimodal function of this protein. Hence it is important to explore this pathway at the molecular level. Two types of trimers, Trimer A (tA) and Trimer B (tB), can be derived from the dodecamer due to the inherent 3-fold symmetry of the spherical crystal structure. These derived trimers were expressed as protein structure graphs (PSGs) using the computed interaction strength among the residues. Interface clusters which were identified from PSGs allowed us to convincingly predict E146 and F47 for further mutation studies. Various single and double mutants were constructed and characterized. We were finally able to generate a single mutant F47E impaired in dodecamerization and a double mutant E146AF47E as native monomer in solution. These two observed results suggest that the two trimers are important for dodecamerization and that the residues selected are important for the structural stability of the protein in vitro.

Identification and characterization of the dps promoter of Mycobacterium smegmatis: promoter recognition by stress-specific extracytoplasmic function sigma factors sigmaH and sigmaF.

The survival of a bacterium with a depleted oxygen or nutrient supply is important for its long-term persistence inside the host under stressful conditions. We studied a gene, dps, from Mycobacterium smegmatis, encoding a protein, Dps (for DNA binding protein from starved cells), which is overexpressed under oxidative and nutritional stresses and provides bimodal protection to the bacterial DNA. Characterization of the dps promoter in vivo is therefore important. We cloned a 1-kb putative promoter region of the dps gene of M. smegmatis in an Escherichia coli-Mycobacterium shuttle vector, pSD5B, immediately upstream of the lacZ gene. Promoter activities were assayed in vivo both in solid medium and in liquid cultures by quantitative beta-galactosidase activity measurements. To characterize the minimal promoter region, a 200-bp fragment from the whole 1-kb sequence was further cloned in the same vector, and in a similar way, beta-galactosidase activity was quantitated. Primer extension analysis was performed to determine the +1 transcription start site of the gene. Point mutations were inserted in the putative promoter sequences in the -10 and -20 regions, and the promoter sequence was confirmed. The promoter was not recognized by purified M. smegmatis core RNA polymerase reconstituted with purified Mycobacterium tuberculosis sigmaA or sigmaB during multiple- and single-round in vitro transcription assays. Promoter-specific in vivo pull-down assays with an immobilized 1-kb DNA fragment containing the dps promoter established that extracellular function sigma factors were associated with this starvation-inducible promoter. Single-round transcription at the dps promoter further supported the idea that only core RNA polymerase reconstituted with sigmaF or sigmaH can generate proper transcripts.

Estimation of Förster's distance between two ends of Dps protein from mycobacteria: distance heterogeneity as a function of oligomerization and DNA binding.

Dps protein (DNA binding Protein from Starved Cells) from Mycobacterium smegmatis (Ms-Dps) is known to undergo an in vitro irreversible oligomeric transition from trimer to dodecamer. This transition helps the protein to provide for bimodal protection to the bacterial DNA from the free radical and Fenton mediated damages in the stationary state. The protein exists as a stable trimer, when purified from E. coli cells transformed with an over-expression plasmid. Both trimer as well as dodecamer are known to exhibit ferroxidation activity, thus removing toxic hydroxyl radicals in vivo, whereas iron accumulation and non-sequence specific DNA binding activity are found in dodecamer only. This seems to be aided by the positively charged long C-terminal tail of the protein. We used frequency domain phase-modulation fluorescence spectroscopy and Förster Resonance Energy Transfer (FRET) to monitor this oligomeric switch from a trimer to a dodecamer and to elucidate the structure of DNA-Dps dodecamer complex. As Ms-Dps is devoid of any Cysteine residues, a Serine is mutated to Cysteine (S169C) at a position adjacent to the putative DNA binding domain. This Cysteine is subsequently labeled with fluorescent probe and another probe is placed at the N-terminus, as crystal structure of the protein reveals several side-chain interactions between these two termini, and both are exposed towards the surface of the protein. Here, we report the Förster's distance distribution in the trimer and the dodecamer in the presence and absence of DNA. Through discrete lifetime analysis of the probes tagged at the respective regions in the macromolecule, coupled with Maximum Entropy Method (MEM) analysis, we show that the dodecamer, upon DNA binding shows conformational heterogeneity in overall structure, perhaps mediated by a non-specific DNA-protein interaction. On the other hand, the nature of DNA-Dps interaction is not known and several models exist in literature. We show here with the help of fluorescence anisotropy measurements of labeled DNA having different length and unlabeled native dodecameric protein that tandem occupation of DNA binding sites by a series of Dps molecules perhaps guide the tight packing of Dps over DNA backbone.