Microbial population and biochemical changes in fermenting finger millet (Eleusine coracana)

Natural fermentation of finger millet (Eleusine coracana) was carried out for 48 h. Microbiological and chemical analysis was performed throughout the fermentation process. The fermentation was heterolactic dominated by lactic acid bacteria accompanied by the production of lactic and acetic acids with decrease in pH and increase in titratable acidity. The microbial population increased until 18 to 24 h accompanied by a rapid decrease in total and reducing sugars. The microflora stabilized between 24 and 48 h, during which time the total and -amylase activities increased with accumulation of sugars. Total free amino acids also increased. Yeast counts were low and moulds and coliforms were absent. Repeated fermentations showed consistency in the qualitative and quantitative changes in microflora. Five predominant types of bacteria, strains belonging to Leuconostoc, Pediococcus and Lactobacillus were identified. Of these only one type, Pediococcus, dominated (>80%) in the latter half of fermentation.     

Host genotype dependency and growth enhancing ability of VA-mycorrhizal fungi for Eleusine coracana (finger millet)

Inoculation of finger millet (Eleusine coracana Gaertn.) plants with one of six different vesicular, arbuscular, mycorrhizal (VAM) fungi increased plant biomass, height, leaf area and absolute growth rate; however, effectiveness of the various VAM fungi varied significantly. Maximum root colonization and mycorrhizal efficacy was observed with plants inoculated with Glomus caledonicum. Among five host genotypes tested for mycorrhizal dependency against G. caledonicum, genotype HR-374 gave the highest plant biomass, mycorrhizal efficacy and root colonization, the inoculation resulting in increased mineral (phosphate, nitrogen, Zn²⁺ and Cu²⁺) content and uptake in shoots.     

In vivo effect of whole grain flour of finger millet (Eleusine coracana) and kodo millet (Paspalum scrobiculatum) on rat dermal wound healing

Influence of finger millet and kodo millet on rat dermal wound healing was assessed by making a 4 cm2 (2 x 2 cm) excision wound on the shaven back of rats under ether anesthesia. Finger millet or kodo millet flour (300 mg) as aqueous paste was applied topically once daily for 16 days. The granulation tissue formed on day 4, 8 and 12 was used to estimate some biochemical parameters like protein, DNA, collagen and lipid peroxides. There was significant increase in protein and collagen contents and decrease in lipid peroxides. Biophysical parameters like rate of contraction and number of days for epithelialization were also studied. Rate of contraction was 88-90% in kodo millet and finger millet treated rats in comparison to 75% in untreated rats. The number of days for complete closure of wounds was lower for finger millet (13 days) and kodo millet (14 days) treated rats in comparison to untreated (16 days) rats. The results implicate a possible therapeutical role for finger millet and kodo millet in accelerating the process of wound healing.     

Development and molecular characterization of genic molecular markers for grain protein and calcium content in finger millet (Eleusine coracana (L.) Gaertn.)

Finger millet (Eleusine coracana (L.) Gaertn), holds immense agricultural and economic importance for its high nutraceuticals quality. Finger millets seeds are rich source of calcium and its proteins are good source of essential amino acids. In the present study, we developed 36 EST-SSR primers for the opaque2 modifiers and 20 anchored-SSR primers for calcium transporters and calmodulin for analysis of the genetic diversity of 103 finger millet genotypes for grain protein and calcium contents. Out of the 36 opaque2 modifiers primers, 15 were found polymorphic and were used for the diversity analysis. The highest PIC value was observed with the primer FMO2E33 (0.26), while the lowest was observed FMO2E27 (0.023) with an average value of 0.17. The gene diversity was highest for the primer FMO2E33 (0.33), however it was lowest for FMO2E27 (0.024) at average value of 0.29. The percentage polymorphism shown by opaque2 modifiers primers was 68.23 %. The diversity analysis by calcium transporters and calmodulin based anchored SSR loci revealed that the highest PIC was observed with the primer FMCA8 (0.30) and the lowest was observed for FMCA5 (0.023) with an average value of 0.18. The highest gene diversity was observed for primer FMCA8 (0.37), while lowest for FMCA5 (0.024) at an average of 0.21. The opaque2 modifiers specific EST-SSRs could able to differentiate the finger millet genotypes into high, medium and low protein containing genotypes. However, calcium dependent candidate gene based EST-SSRs could broadly differentiate the genotypes based on the calcium content with a few exceptions. A significant negative correlation between calcium and protein content was observed. The present study resulted in identification of highly polymorphic primers (FMO2E30, FMO2E33, FMO2-18 and FMO2-14) based on the parameters such as percentage of polymorphism, PIC values, gene diversity and number of alleles.     

Efficiency of RAPD, SSR and Cytochrome P450 gene based markers in accessing genetic variability amongst finger millet (Eleusine coracana) accessions

Finger millet (Eleusine coracana L.) is an important crop used for food, forage, and industrial products. Three DNA marker techniques, random amplified polymorphic DNA (RAPD), simple sequence repeat (SSR) and cytochrome P₄₅₀ gene based markers were used for the detection of genetic polymorphism in 83 accessions of finger millet collected from various geographical regions of India and Africa. A total of 18 RAPD, 10 SSR and 10 pairs of cytochrome P₄₅₀ gene based markers were generated 56.17, 70.19 and 54.29% polymorphism, respectively. Mean polymorphism information content (PIC) for each of these marker systems (0.280 for RAPD, 0.89 for SSR and 0.327 for cytochrome P₄₅₀ gene based markers) suggested that SSR marker were highly effective in determining polymorphism. The phenograms based on the three markers data indicate that genotypes from different geographical regions are clearly distinguishable as separate clusters. Mantel test employed for detection of goodness of fit established cophenetic correlation values above 0.90 for all the three marker systems. The dendrograms and PCA plots derived from the binary data matrices of the three marker systems are highly concordant. High bootstrap values were obtained at major nodes of phenograms through WINBOOT software. Based on the results of present study, SSR and cytochrome P₄₅₀ gene based markers appear to be particularly useful for the estimation of genetic diversity. This study reveals the potential of RAPD, SSR and gene based markers for characterizing germplasm of Eleusine coracana and narrow down the vast germplasm into distinct core groups.     

Effect of sulfur supplements on cellulolytic rumen micro-organisms and microbial protein synthesis in cattle fed a high fibre diet

AIM: To examine the effect of sulfur-containing compounds on the growth of anaerobic rumen fungi and the fibrolytic rumen bacteria Ruminococcus albus, Ruminococcus flavefaciens and Fibrobacter succinogenes in pure culture and within the cattle rumen. METHODS AND RESULTS: The effect of two reduced sulfur compounds, 3-mercaptopropionic acid (MPA) or 3-mercapto-1-propanesulfonic acid as the sole S source on growth of pure fibroyltic fungal and bacterial cultures showed that these compounds were capable of sustaining growth. An in vivo trial was then conducted to determine the effect of sulfur supplements (MPA and sodium sulfate) on microbial population dynamics in cattle fed the roughage Dichanthium aristatum. Real-time PCR showed significant increases in fibrolytic bacterial and fungal populations when cattle were supplemented with these compounds. Sulfate supplementation leads to an increase in dry matter intake without a change in whole tract dry matter digestibility. CONCLUSIONS: Supplementation of low S-containing diets with either sodium sulfate or MPA stimulates microbial growth with an increase in rumen microbial protein supply to the animal. SIGNIFICANCE AND IMPACT OF THE STUDY: Through the use of real-time PCR monitoring, a better understanding of the effect of S supplementation on discrete microbial populations within the rumen is provided.     

Effect of ambient temperature on nutrient digestibility and nitrogen balance in sheep fed brown-midrib maize silage

The aim of the experiment was to determine the impact of heat stress on nutrient digestibility and nitrogen balance in sheep fed silages differing in fibre quality. The digestibility trial was conducted at three different ambient temperatures (15°C, 25°C and 35°C for 24 h/d). The tested brown-midrib maize (Bm) silage had a higher nutrient digestibility, except for ether extract (EE) and a higher metabolisable energy (ME) content than the control maize (Con) silage. Nitrogen (N) excretion with faeces was higher but N excretion with urine was lower for sheep fed Bm silage, subsequently N balance did not differ between the two silages. Temperature had no effect on nutrient digestibility, except for crude protein (CP), but N excretion with urine was lower at elevated temperatures. A diet by temperature interaction was found for dry matter (DM) and organic matter (OM) digestibility. When the ambient temperature increased from 15°C to 25°C, the DM and OM digestibility increased in animals fed Con silage, but decreased in animals fed Bm silage. Concomitantly, ME estimated from digestible nutrients was higher for Bm than for Con at 15°C, but no differences were found at 25°C and 35°C. Effects of diet by temperature interaction, furthermore, were observed for EE and CP digestibility. Therefore, forage quality has to be considered when feeding heat-stressed animals.     

Diversity of rumen microbiota using metagenome sequencing and methane yield in Indian sheep fed on straw and concentrate diet

An in vivo study aiming to investigate the rumen methanogens community structure was conducted in Mandya sheep fed on straw and concentrate diet. The ruminal fluid samples were collected and processed for unravelling the rumen microbiota and methanogens diversity. Further, the daily enteric methane emission and methane yield was also quantified using the SF₆ tracer technique. Results indicated that the Bacteroidetes (～57%) and Firmicutes (25%) were two prominent affiliates of the bacterial community. Archaea represented about 2.5% of the ruminal microbiota. Methanobacteriales affiliated methanogens were the most prevalent in sheep rumen. The study inveterate that the ruminal archaea community in sheep is composed of 9 genera and 18 species. Methanobrevibacter represented the largest genus of the archaeome, while methylotrophs genera constituted only 13% of the community. Methanobrevibacter gottschalkii was the prominent methanogen, and Methaobrevibacter ruminantium distributed at a lower frequency (～2.5%). Among Methanomassiliicoccales, Group 12 sp. ISO4-H5 constituted the most considerable fraction (～11%). KEGG reference pathway for methane metabolism indicated the formation of methane through hydrogenotrophic and methylotrophic pathways, whereas the acetoclastic pathway was not functional in sheep. The enteric methane emission and methane yield was 19.7 g/d and 20.8 g/kg DMI, respectively. Various species of Methanobrevibacter were differently correlated, and the distribution of hydrogenotrophic methanogens mainly explained the variability in methane yield between the individual sheep. It can be inferred from the study that the hydrogenotrophic methanogens dominate the rumen archaeal community in sheep and methylotrophic/aceticlastic methanogens represent a minor fraction of the community. Further studies are warranted for establishing the metabolic association between the prevalent hydrogenotrophs and methylotrophs to identify the key reaction for reducing methane emission.     

Effect of suplemental nitrogen source and feeding frequency on nutrient supply to lambs fed a kikuyu grass (Pennisetum clandestinum) hay-based diet

Eight castrated male lambs (35 ± 4 kg live weight), fed a basal diet of kikuyu grass hay, were used in a replicated 4 × 4 Latin Square experiment with a 2 × 2 factorial arrangement of treatments to evaluate the effect of supplemental feeding frequency and source of rumen degradable N on intake, digestibility, ruminal fermentation, and microbial protein yield. Treatments were supplementation with cassava meal plus calcium caseinate or cassava meal plus urea offered at a rate of 7 g/kg live weight daily in one or two meals per day. Lambs were fed twice daily in such manner to allow ad libitum comsumption of forage. There was significant feeding frequency by N source interaction on variables of intake. In general, intake of feed components was higher (P ≤ 0.05) by lambs offered the caseinate-supplement twice daily over intake observed in lambs given the others diet treatments. Digestibility of feed components was neither affected by supplemental N source (DM, P = 0.541; OM, P = 0.585; NDF, P = 0.828) nor by feeding frequency (DM, P = 0.122; OM, P = 0.175; NDF, P = 0.591). Urinary excretion of N increased (P ≤ 0.05) in lambs supplemented twice daily whereas N retention was similar for all treatments (N source, P = 0.748; feeding frequency, P = 0.418). Microbial protein entering into the small intestine was affected by the interaction between feeding frequency and N source such as an increasing (P < 0.10) in this variable was observed when lambs received the caseinate but not the urea supplement twice daily. Efficiency of microbial protein synthesis, however, was not affected by treatments (N source, P = 0.588; feeding frequency, P = 0.334). Rumen pH averaged 6.70 and it was neither affected by N source (P = 0.827) nor by feeding frequency (P = 0.740). Ruminal concentration of ammonia N was not affected by feeding frequency (P = 0.144) while it increased (P < 0.05) when urea rather than caseinate was the supplemental N source (mean of 7.61 mg/dl vs. 6.00 mg/dl). Concentration of sugars in rumen fluid was higher (P ≤ 0.05) in lambs supplemented once a day compared to twice daily (mean of 49.4 mg/dl vs. 34.4 mg/dl) for both N sources. A significant (P ≤ 0.05) N source by feeding frequency interaction effect was observed for ruminal concentrations of α-amino N compounds. In urea treatment α-amino N concentration increased (P ≤ 0.05) in lambs receiving the supplement twice daily compared to once a day (mean of 4.59 mg/dl vs. 3.70 mg/dl) while in caseinate treatment it was higher (P ≤ 0.05) in lambs offered the supplement in one meal per day compared to twice daily (mean of 5.29 mg/dl vs. 4.07 mg/dl). In conclusion, for ruminants fed a tropical grass-based diet, starch-rich supplement containing non-protein N as N source may be offered only once a day whereas the supply of nutrients may be improved if degradable true protein is included as N source and supplement is offered in two meals per day.     

Effect of individual and mixed live yeast culture feeding on growth performance,nutrient utilization and microbial crude protein synthesis in lambs

This study investigated effects of feeding three individual, and a mixed, yeast culture (Kluyveromyces marximanus NRRL3234, Saccharomyces cerevisiae NCDC42, Saccharomyces uvarum ATCC9080 all in a 1:1:1, ratio) on growth performance, nutrient utilization and microbial crude protein (CP) synthesis in feedlot lambs during the post-weaning phase of growth. Sixty weaner lambs (90 ± 3.5 d old and 15.9 ± 0.50 kg BW) were fed for 91 d in five equal groups. The control group of lambs received sterilized culture medium while the treatment groups were fed a yeast culture in addition to a ad libitum total mixed ration (TMR). The yeast culture, dosed at 1 ml/kg body weight (BW) had 1.5–2.0 × 10⁹ live cells/ml. Yeast culture supplementation did not influence intake and digestibility of organic matter (OM), CP, neutral detergent fiber (NDF), acid detergent fiber (ADF) and hemicellulose and the metabolizable energy (ME) level of the diets were similar between control and yeast supplemented lambs. Lambs in all groups were in positive N balance, but N intake and N voided in feces and urine, as well as N balance, did not change due to yeast culture supplementation. Urinary allantoin excretion was similar, but purine derivatives absorbed (mM/d) were higher (P<0.05) in yeast culture supplemented lambs. Yeast culture supplementation improved (P<0.05) microbial CP synthesis. Supplementation of SC and mixed yeast improved (P=0.002) BW gain of lambs by 21% and 16% respectively. All yeast culture supplemented lambs had higher feed efficiency in comparison to control lambs. Among the three yeast cultures used, S. cerevisiae had the most potential as a growth promoting feed additive in feedlot lamb production, and it may serve as an alternate to antibiotics and ionophores as a growth promoter of weaner lambs.     

Replacing soybean meal with flax seed meal: effects on nutrient digestibility,rumen microbial protein synthesis and growth performance in sheep

Flax seed meal (FSM) is rich in various nutrients, especially CP and energy, and can be used as animal protein feed. In animal husbandry production, it is a long-term goal to replace soybean meal (SBM) in animal feed with other plant protein feed. However, studies on the effects of replacing SBM with FSM in fattening sheep are limited. The aim of this experiment was to study the effects of replacing a portion of SBM with FSM on nutrient digestibility, rumen microbial protein synthesis and growth performance in sheep. Thirty-six Dorper × Small Thin-Tailed crossbred rams (BW = 40.4 ± 1.73 kg, mean ± SD) were randomly assigned into four groups. The dietary treatments (forage/concentrate, 45 : 55) were isocaloric according to the nutrient requirements of rams. Soybean meal was replaced with FSM at different levels (DM basis): (1) 18% SBM (18SBM), (2) 12% SBM and 6% FSM (6FSM), (3) 6% SBM and 12% FSM (12FSM) and (4) 18% FSM (18FSM). The rams were fed in individual pens for 60 days, with the first 10 days for adaptation to diets, and then the digestibility of nutrients was determined. There was no significant difference in DM intake, but quadratic (P < 0.001) effects on the average daily gain and feed efficiency were detected, with the highest values in the 6FSM and 12FSM groups. For DM and NDF digestibility, quadratic effects were observed with the higher values in the 6FSM and 12FSM groups, but the digestibility of CP linearly decreased with the increase in FSM in the diet (P = 0.043). There was a quadratic (P < 0.001) effect of FSM inclusion rate on the estimated microbial CP yield. However, the values of intestinally absorbable dietary protein decreased linearly (P < 0.001). For the supply of metabolisable protein, both the linear (P = 0.001) and quadratic (P = 0.044) effects were observed with the lowest value in the 18FSM group. Overall, the results indicated that SBM can be effectively replaced by FSM in the diets of fattening sheep and the optimal proportion was 12.0% under the conditions of this experiment.     

Amino acid metabolism and protein synthesis in lactating rats fed on a liquid diet.

1. Amino acid metabolism was studied in control virgin rats, lactating rats and virgin rats protein-pair-fed with the lactating rats (high-protein virgin rats). 2. Urinary excretion of nitrogen and urea was higher in lactating than in control virgin rats, and in high-protein virgin rats it was higher than in lactating rats. 3. The activities of urea-cycle enzymes (units/g) were higher in high-protein virgin than in lactating rats, except for arginase. In lactating rats the activities of carbamoyl-phosphate synthase, ornithine carbamoyltransferase and argininosuccinate synthase were lower than in control virgin rats. When the liver size is considered, the activities in lactating rats were similar to those in high-protein virgin rats, except for arginase. 4. N-Acetylglutamate content was higher in high-protein virgin rats than in the other two groups. 5. The rate of urea synthesis from precursors by isolated hepatocytes was higher in high-protein virgin rats than in the other two groups. 6. The flooding-dose method (L-[4-3H]phenylalanine) for measuring protein synthesis was used. The absolute synthesis rates of mammary gland, liver and small-intestinal mucosa were higher in lactating rats than in the other two groups, and in high-protein virgin rats than in control virgin rats 7. These results show that the increased needs for amino acids during lactation are met by hyperphagia and by a nitrogen-sparing mechanism.     

Effects of sun-drying and exogenous enzymes on nutrients intake,digestibility and nitrogen utilization in sheep fed Atriplex halimus foliages

This study was conducted to assess effects of sun-drying and/or addition of an exogenous enzyme (ENZ) preparation on intake, digestibility of nutrients and recovery values of secondary metabolites (SM) in the gastrointestinal tract of sheep fed Atriplex halimus (AH) foliages. A randomized block design for 28 d was used for four experimental treatments based on either fresh (AH-F) or sun-dried (AH-S) A. halimus foliages in the absence (?ENZ) or presence (+ENZ) of 10 g/sheep/d of the exogenous of ZADO^® enzyme preparation. Three adult sheep weighing 51 ± 2.7 kg were fed for each experimental treatment. The ENZ was added daily with a small amount of concentrate to help balance the dietary metabolizable energy concentration. Nutrient intake and digestibility, N balance and recovery of SM (i.e., total phenolics (TP), saponins (SP), alkaloids (AK), aqueous fraction (AF)) in the gastrointestinal tract were determined. Levels of most nutrients did not differ between AH-F and AH-S foliages, but the AH-S contained less than half of the SM in AH-F. Drying of A. halimus and ENZ addition increased (P=0.001) intake as well as OM and NDFom digestibility (P=0.02). Feed intake and digestibility were higher (P=0.01) in AH-S with ENZ addition. Intake of N by sheep fed the treatment diets depended on DM intake as the dietary concentration of N in the diets was similar. Thus AH-S sheep supplemented with ENZ had higher (P=0.001) N intake. Digestibility of N was similar to DM and OM digestibility, and was higher (P=0.03) in AH-S sheep supplemented with ENZ. Drying and ENZ addition to the diet increased (P=0.004) recovery of all SM. The fate of these compounds in the rumen needs to be evaluated considering that SM have been implicated in fiber and protein degradation in the rumen. The study showed that there are beneficial impacts of sun-drying and/or dietary exogenous enzyme addition for sheep fed A. halimus.     

Substitution of finger millet (Eleusine coracana) and bulrush millet (Pennisetum typhoides) for maize in broiler feeds

The first experiment was conducted to determine acceptability levels and growth performance when bulrush millet or finger millet was substituted for maize at 0, 20, 40 or 60% in both broiler starter and finisher feeds. The chicks fed on the bulrush millet diets had the highest overall body weight gain of 1649 g per chick at 55 days. Performance on the finger millet diets was comparable to that on maize. In view of the higher protein content in the bulrush millet tested, subsequent experiments were conducted to determine the value of using bulrush millet not only as an energy source, but also as a part substitute for protein supplements in broiler diets. The bulrush millet included at 70% was shown to effectively replace part of the vegetable protein supplement provided the diet was supplemented with up to 0.3% lysine.     

Use of genomic and genic SSR markers for assessing genetic diversity and population structure in Indian and African finger millet (Eleusine coracana (L.) Gaertn.) germplasm

Genetic diversity and population structure were analyzed in 67 diverse finger millet accessions of African and Indian origin. A total of 69 alleles were generated with a mean of 4.0 alleles per locus and with an average gene diversity of 0.471. Molecular diversity parameters showed higher values in African accessions. Nineteen rare and nine unique alleles were observed and the identified accessions can be a potential source for further improvement of finger millet. Clustering of south Indian accessions with African lowland types and north/highland Indian accessions with that of African highlands was also observed. Structure analysis revealed the distinctness of Ugandan accessions compared to other African accessions and five major sub-populations useful for parental selection, conservation, and utilization of finger millet.     

Variation in response of accessions of minor millets,Pennisetum americanum (L.) Leeke (pearl millet) and Eleusine coracana (L.) Gaertn (finger millet), and Eragrostis tef (Zucc.) Trotter (tef) to salinity in early seedling growth

The response to increasing NaCl concentration of seedlings of 25 accessions of Ethiopian land races of each of Pennisetum americanum (L.) Leeke (pearl millet) and Eleusine coracana (L.) Gaertn (finger millet), and 15 accessions of Eragrostis tef (Zucc.) Trotter (tef), was examined after two week's growth in NaCl solution culture. Although increasing NaCl concentration significantly reduced seedling root lengths, there was considerable variation within, and between accessions within each species.Analysis based upon a non-linear least square inversion method, using root length data, revealed significant differences in accessions of P. americanum and E. tef on the basis of the estimated salinity threshold, C _t , the NaCl concentrations at which root length begins to decrease. C _t did not differ significantly between E. coracana accessions. Estimates of C₅₀ and C₀, mininum concentrations causing a 50% decrease in root length, and zero root growth respectively, revealed differences between and within accessions for all three species. Overall, finger millet was more tolerant than tef, which was more tolerant than pearl millet. There is clear evidence that differences in tolerance are genetically based from broad sense heritability estimates.     

Supplementation with non-fibrous carbohydrates reduced fiber digestibility and did not improve microbial protein synthesis in sheep fed fresh forage of two nutritive values

To determine whether non-fibrous carbohydrate (NFC) supplementation improves fiber digestibility and microbial protein synthesis, 18 Corriedale ewes with a fixed intake level (40 g dry matter (DM)/kg BW0.75) were assigned to three (n = 6) diets: F = 100% fresh temperate forage, FG = 70% forage + 30% barley grain and FGM = 70% forage + 15% barley grain + 15% molasses-based product (MBP, Kalori 3000). Two experimental periods were carried out, with late (P1) and early (P2) vegetative stage forage. For P2, ewes were fitted with ruminal catheters. Forage was distributed at 0900 h, 1300 h, 1800 h and 2300 h, and supplement added at 0900 h and 1800 h meals. Digestibility of the different components of the diets, retained N and rumen microbial protein synthesis were determined. At the end of P2, ruminal pH and N-NH3 concentration were determined hourly for 24 h. Supplementation increased digestibility of DM (P < 0.001) and organic matter (OM; P < 0.001) and reduced NDF digestibility (P = 0.043) in both periods, with greater values in P2 (P = 0.008) for the three diets. Daily mean ruminal pH differed (P < 0.05) among treatments: 6.33 (F), 6.15 (FG) and 6.51 (FGM). The high pH in FGM was attributed to Ca(OH)2 in MBP. Therefore, the decreased fiber digestibility in supplemented diets could not be attributed to pH changes. The mean ruminal concentration of N-NH3 was 18.0 mg/dl, without differences among treatments or sampling hours. Microbial protein synthesis was greater in P2 (8.0 g/day) than in P1 (6.1 g/day; P = 0.006), but treatments did not enhance this parameter. The efficiency of protein synthesis tended to be lower in supplemented groups (16.4, 13.9 and 13.4 in P1, and 20.8, 16.7 and 16.2 g N/kg digestible OM ingested in P2, for F, FG and FGM, respectively; P = 0.07) without differences between supplements. The same tendency was observed for retained N: 2.55, 1.38 and 1.98 in P1, and 2.28, 1.23 and 1.10 g/day in P2, for F, FG and FGM, respectively; P = 0.05). The efficiency of microbial protein synthesis was greater in P2 (P = 0.007). In conclusion, addition of feeds containing NFCs to fresh temperate forage reduced the digestibility of cell walls and did not improve microbial protein synthesis or its efficiency. An increase in these parameters was associated to the early phenological stage of the forage.     

Isolation of a cDNA clone (PcSrp) encoding serine-rich-protein from Porteresia coarctata T. and its expression in yeast and finger millet (Eleusine coracana L.) affording salt tolerance

A 1.4 Kb cDNA clone encoding a serine-rich protein has been isolated from the cDNA library of salt stressed roots of Porteresia coarctata, and designated as P. coarctata serine-rich-protein (PcSrp) encoding gene. Northern analysis and in situ mRNA hybridization revealed the expression of PcSrp in the salt stressed roots and rhizome of P. coarctata. However, no such expression was seen in the salt stressed leaves and in the unstressed tissues of root, rhizome and leaf, indicating that PcSrp is under the control of a salt-inducible tissue-specific promoter. In yeast, the PcSrp conferred increased NaCl tolerance, implicating its role in salinity tolerance at cellular level. Further, PcSrp was cloned downstream to rice Actin-1 promoter and introduced into finger millet through particle-inflow-gun method. Transgenic plants expressing PcSrp were able to grow to maturity and set seed under 250 mM NaCl stress. The untransformed control plants by contrast failed to survive under similar salt stress. The stressed roots of transgenic plants invariably accumulated higher Na⁺ and K⁺ ion contents compared to roots of untransformed plants; whereas, shoots of transgenics accumulated lower levels of both the ions than that of untransformed plants under identical stress, clearly suggesting the involvement of PcSrp in ion homeostasis contributing to salt tolerance.     

Effectiveness of phytase pre-treatment on growth performance,nutrient digestibility and mineral status of common carp (Cyprinus carpio) juveniles fed Moringa by-product based diet

Several anti-nutritional substances are found in plant derivatives for example phytate, that make the nutrients and minerals unavailable to fish, hence leading to poor growth performance. Presence of the anti-nutrient factor such as phytate is a chelated compound and need enzyme for its breakdown and availability of nutrients to improve fish growth. This research work was performed to check the improvement of overall performance of Cyprinus carpio fingerlings by the help of phytase addition in Moringa oleifera by- products based diet. Combination of Moringa seed meal and Moringa leaf meal was utilized as test ingredient to formulate seven test feeds, containing graded levels of phytase (0, 500, 650, 800, 950, 1100 and 1250 FTU kg⁻¹). In feeding trial of 70 days, fingerlings were given feed two times in a day at the rate of 4% of wet weight of their bodies and faeces were collected. According to current results, it was found that growth performance parameters i.e. weight gain; 25 g, specific growth rate; 1.67 and feed conversion ratio; 1.10 were improved to maximum at 950 FTU kg⁻¹. Digestibility of nutrients (crude protein; 73%, crude fat; 71% and gross energy; 67%) and minerals absorption was also maximum (Ca; 70%, Zn; 66%, K; 74%, Mn; 66% and P; 71%) at 950 FTU kg⁻¹. Lowest growth efficiency, nutrient digestibility and mineral absorption were observed in fingerlings fed at control diet (0 FTU kg⁻¹). Results of the current study, proved that 950 FTU kg⁻¹ is the most optimum level of phytase to formulate economical and ecofriendly feed for improved growth of C. carpio fingerlings as it decreases the discharge of minerals and nutrients in water bodies.