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A total of 29 species in 16 genera and 8 subfamilies of the braconids (Hym., Braconidae) were collected from alfalfa fields in Bahariya and Farafra oases (Western Desert, Egypt) These species were identified, together with details of the sites where they were found and an indication of their relative abundance in each site. Among the samples, 4 species in 4 genera and 3 subfamilies are recorded as new to the Egyptian fauna. These are: Aphidius asteris Haliday, 1834, Ephedrus plagiator (Nees, 1811) (Aphidiinae), Homolobus (Chartolobus) infumator (Lyle, 1914) (Homolobinae) and Opius lugens Haliday, 1837 (Opiinae). Thirteen braconid species are also newly recorded in association with alfalfa fields, namely A. asteris; Praon necans Mackaeur, 1959 (Aphidiinae); Habrobracon hebetor (Say, 1836) (Braconinae); Chelonus basalis Curtis, 1837; Ch. blackburni Cameron, 1886; Ch. oculator (Fabricius, 1775); Phanerotoma leucobasis Kriechbaumeri, 1894 (Cheloninae); Dinocampus coccinellae (Schrank, 1802) (Euphorinae); H. (C.) infumator (Lyle, 1914) (Homolobinae); Apanteles appellator Telenga, 1949; Cotesia glomerata (Linnaeus, 1758) (Microgastrinae); Psyttalia concolor (Szépligeti, 1910) and P. nilotica (Schmiedeknechti, 1900) (Opiinae).

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Background: Concentrations of 4 toxic metals, viz. Cd, Cu, Pb, and Zn in the soil and alfalfa samples collected from Borujerd, Iran, was determined. The capability of alfalfa to accumulate heavy metals from soils was assessed in terms of Biological Concentration Factor. Materials and Methods: The alfalfa and soil samples were collected from 20 different farms, including 13 wastewater-irrigated and seven underground-irrigated farms. After acid digestion, the samples were analyzed using atomic absorption spectrophotometer. Results: The levels of Cd, Pb, and Zn in the soils of wastewater-irrigated farms were higher than those from the groundwater-irrigated farms. With the exception of Cu, concentrations of heavy metals in the alfalfa crop were higher in wastewater-irrigated farms compared to well water. Also, in the case of BCF, both Cd and Cu values decreased with increasing metal concentration in soil. The order of BCF of heavy metals in alfalfa was in order of Cu>Cd>Zn>Pb in well water-irrigated and Zn>Cd>Cu>Pb in wastewater –irrigated samples. Discussion and Conclusions: The findings remarked that the levels of Cu, Cd, and Pb in alfalfa were exceeding the permissible levels suggested by the Joint FAO/WHO Expert Committee on Food Additives. These outcomes propose that the consumption of alfalfa plants is potentially threatening both animal and human health.

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Population dynamics of three main alfalfa aphids, namely: pea aphid, Acyrthosiphon pisum (Harris), spotted alfalfa aphid, Therioaphis trifolii forma maculata (Buckton), black alfalfa aphid, Aphis craccivora Koch, and their most abundant predators and parasitoids in three sites and in two subsequent years in Isfahan, Iran, were investigated. The results revealed that frequency and temporal occurrence of alfalfa aphids and their natural enemies in different regions were varied, but aphid natural enemy populations were more or less coincident with aphid populations. Populations of aphids were mainly affected by alfalfa harvesting, ambient temperature, and coccinellid predators. Hemipterous predators in contrast with coccinellids, had little effect on aphid populations. Parasitoids seem to be effective on low populations of alfalfa aphids. Air humidity and rainfall had no effect on populations of alfalfa aphids.

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Enzyme electrophoresis was used to measure genetic variation within, and divergence among, three generations of recently bred synthetic alfalfa generations (Syn1, Syn2, and Syn3) originating from a polycross of 12 selected parents and several cultivars. Three isozyme loci, exhibiting tetrasomic inheritance in 10-day seedlings, were detected from five enzymatic systems analyzed by polyacrylamide slab gel electrophoresis for about 100 individuals of each alfalfa population. Very high levels of heterozygosity (ranging from 0.521 to 0.699) were observed within alfalfa populations, using polymorphic loci. The reduction in heterozygosity was about 5% from Syn1 to Syn2 and from Syn2 to Syn3. The last open pollinated generation was found to be in W-H equilibrium as well as Gharayonja, a native ecotype under examination, using c2-test. Application of Wright's Fstatistics revealed that the estimated overall inbreeding coefficient, (FIT), of 9.4% was mainly related to inbreeding or double reduction in alfalfa (FIS= 8.61%) rather than random genetic drift or population differentiation (FST= 1.6%). Therefore, due to very large intra-population diversity, the breeding program of the synthetic alfalfa did not generate a large variety differentiation. However, the use of seedling allozymic loci can be applied successfully for estimation of the population genetic parameters.

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Physical and mechanical properties of alfalfa (Medicago sativa, L.) grind are required for optimum design of equipment used in transporting, processing and storage of the material. This study was conducted to determine the effect of particle size (2.38, 3.36 and 4.76 mm) and moisture content (8, 9.3 and 11% wb) on some physical and mechanical properties of alfalfa grind. These properties include: geometric mean diameter, bulk density, coefficient of static friction (on galvanized iron, Plexiglass, rubber and polished steel surfaces), filling angle of repose, coefficient of internal friction, cohesion, and adhesion to polished steel plate. The bulk density varied from 161.6 to 202.2 kg m-3. The coefficient of static friction changed from 0.26 on polished steel plate to 0.87 on rubber surface. Larger particles with higher moisture content had the highest filling angle of repose (54.5°). The coefficient of internal friction varied from 0.64 to 0.88. The 2.38-mm alfalfa grind at moisture content of 11% (wb), and the 4.76-mm at moisture content of 8% (wb) had the highest and lowest cohesion (7.65 and 4.80 kPa), respectively. The adhesion on polished steel plate varied from 0.19 to 1.54 kPa.

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Alfalfa crops were surveyed for the incidence of alfalfa mosaic virus (AMV), cucumber mosaic virus (CMV), peanut stunt virus (PSV), bean leaf roll virus (BLRV), bean yellow mosaic virus (BYMV) and bean common mosaic virus (BCMV) in the major growing areas in the southeast and central regions of Iran. Samples were collected between May 2009 and March 2011 and analyzed for viral infection initially by enzyme-linked immunosorbent assay (ELISA) followed by RT-PCR using capsid protein gene specific primers. In total, 634 symptomatic leaf samples were collected in four southeastern and central provinces of Iran representing 20 regions. Our results revealed a high incidence of AMV over a wide geographical area. AMV and BLRV were identified in most regions, whereas BYMV was found only in Yazd Province. PSV was detected in three regions, but not in Sistan- Balouchestan and Hormozgan Provinces. The highest incidence of viral infection amongst the surveyed provinces was recorded in Kerman (66.8%), followed by Yazd (39%), Sistan and Balouchestan (20.8 %), and Hormozgan (4.5%). AMV, BLRV, PSV and BYMV were present in 23.3%, 12%, 0.70% and 0.28% of the samples, respectively. CMV and BCMV were not detected in any surveyed region. Multiple virus infections were recorded in 42 samples. This is the first report on the detected occurrence of BLRV, PSV and BYMV in alfalfa in the southeast and central regions of Iran.

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Salinity is among important soil stresses adversely affecting the process of nitrogen (N) fixation in leguminous plants in different parts of the world. It has been indicated that salinity can inhibit the early stages of nodulation process between bacterium and the host plant including the exchange of signal molecules (nod gene inducers). There has not been any research regarding the effects of nod gene inducers on the growth of alfalfa inoculated with Sinorhizobium meliloti under saline conditions. A growth chamber experiment was conducted to determine the effects of pre-incubation of S. meliloti with effective inducers of nod genes Luteolin, Methyl jasmonate and Genistein on the growth and N-fixation of two different alfalfa (Medicago sativa L.) cultivars (Yazdi and Hamedani) under salt stress. Nod gene inducers increased alfalfa growth and N fixation under normal as well as under salt stressed conditions. Yazdi cultivar showed to be more tolerant to salinity than Hamedani with a higher growth rate and N fixation. Luteolin was the most effective nod gene inducer on plant growth and N fixation under normal and as well under salt stressed conditions. The results suggest that pre-incubation of S. meliloti with effective nod gene inducers can improve alfalfa growth and N fixation under salinity stress.

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 In order to evaluate salt (NaCl) tolerance in alfalfa (Medicago sativa L.) half-sib families, a factorial experiment on the basis of completely randomized design, with 20-25 individuals in pot (replicates) was carried out under field conditions at the Research Station of University of Tabriz. Electrophoretic analyses were performed by using 7.5% slab polyacrylamide gels. Two antioxidant enzymes including superoxide dismutase (SOD) and peroxidase (POX) and one common isozymic system namely esterase (EST) were stained and for each isozymic band the “density × area” scores onto gels were evaluated by MCID software as enzymatic activity. Plant materials consisted of 12 half-sib families that were obtained from a “polycross nursery progeny test” in the same station, a few years ago. The applied salt stress (9±0.2 ds m^-1) reduced plant height, dry weight, leaf weight and stem weight about 31.7, 37.5, 33.7 and 34.7 percent, respectively. Significant correlations were observed between plant height and some antioxidant isozyme activities. Salt stress increased activities of some SOD and POX isozymes but it was ineffective on the activity of EST. Among the families, Ranger cultivar and Zaghl-aghaj and Taze-kand landraces which displayed maximum height and other characteristics exhibited the highest increments in respect of isozyme activities, indicating that antioxidant analysis by gel electrophoresis could be a useful tool for salt stress tolerance studies.

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Iran stands out as a significant center of genetic diversity for alfalfa (Medicago sativa) worldwide, harboring diverse types of this plant. Ensuring the authenticity of alfalfa populations and varieties is crucial for farmers and seed producers, as the genetic makeup of this species directly influences forage and seed yield quality. In this study, we developed a method to identify and differentiate key Iranian cultivated alfalfa populations using microsatellite markers. We collected random samples, each containing 100 seeds, from various alfalfa accessions. Nine microsatellite loci were screened and employed to differentiate these populations based on specific allelic genotypes. Notably, the MTIC233, BI90, ACT009, TC7, MTIC183, MS30, MTIC238, and AFCA11 markers exhibited the highest differentiation ability. The genetic distance analysis revealed that 5-B and foreign accessions, as well as 29-N and foreign accessions, were the most distant from each other. Conversely, 27-G, 9-H, and 21-R exhibited the closest genetic similarity. The results revealed that, accessions 9-H, 21-R, 27-G, 25-B, 5-B, and 2-G shared a common genetic background, suggesting their close relatedness. Our proposed method allows straightforward identification of target alfalfa accessions within a short timeframe (one to two days) without the need for DNA extraction from leaves.

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In this study, the kinetics of total antioxidant activity changes by DPPH radical evaluation, brown pigment formation (BPF), and phenol total with folin–ciocalteu reagent intestinal tract in heated alfalfa honey at different temperatures (45, 55 and 65 °C) over a period of 10 days it placed. The results showed that the amount of antioxidant activity, BPF, and total phenol content increased with increasing temperature and time. Also, the kinetics of changes in BPF and total phenol showed that these parameters follow zero-order kinetics and the activation energy was 86.1 and 71.7 kJ / mol, respectively, at 45-65 °C. However, due to the diversification of antioxidant activity at different temperatures, second order, first order, and zero order kinetics were obtained at 45, 55 and 65 ° C, respectively. Honey heating at 65 ° C was more effective than 45 ° C and 55 ° C for all three parameters. The results showed that antioxidant activity was associated with an increase in both browning factors and total phenol, and also with increasing brown pigment formation, the total phenol content increased so that the highest amount of phenol was related to the darkest honey sample.

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Limited data exist about the production of extruded linseed as a feed ingredient in farm animal nutrition. The objective of this study was to produce and evaluate extruded linseed mixed with alfalfa hay, pistachio by-products, and sugar beet pulp or corn grain in different proportions of linseed: alfalfa hay: pistachio by-products at ratios of 70:15:15, 70:20:10, 70:10:20, and 80:10:10 for quality parameters and in situ rumen Dry Matter (DM) degradability. Using a completely randomized design, the treatment containing alfalfa hay had higher Extrusion Effectiveness (EE), Water Holding Capacity (WHC), and Angle of Repose (AR), but lower Oil Loss (OL) and Bulk Density (BD) than other treatments (P< 0.05). There were no differences among extruded linseed products with different ratios of linseed: alfalfa: pistachio by-products for EE, WHC, and AR (P> 0.05). The treatment with an 80:10:10 ratio had the highest OL among all treatments and the treatment with a 70:15:15 ratio had lower BD than the others (P< 0.05). The DM degradability parameters of extruded products was affected by the absorbents and the treatment containing alfalfa hay had higher potential DM degradability than other treatments (P< 0.05). In conclusion, extruded treatment with alfalfa hay had the highest EE, oil retention capacity and potential rumen DM degradability compared to other treatments. In addition, extruded linseed product with an 80:10:10 ratio of linseed: alfalfa hay: pistachio by-products had higher OL than the other ratios.

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Nowadays, natural pigments are widely used in the food, cosmetics and sanitation industries. In recent years, numerous researches have been done on the methods of extraction and evaluation of the properties of natural pigments. The purpose of this study is comparison of enzymatic and ultrasonic extraction methods for the antioxidant properties of chlorophyll extracted from alfalfa (Medicago sativa L.). Antioxidant activity was performed by three methods including DPPH and ABTS free radicals scavenging and ferric reducing antioxidant power (FRAP). The results show that the concentration of chlorophyll a in alfalfa is higher than that of chlorophyll b and also the enzymatic method demonstrates higher yield in chlorophyll extraction. In addition, higher concentrations of alfalfa extract showed higher antioxidant activity in inhibiting DPPH free radicals and ferric reducing antioxidant power (p <0.05). Also, with increasing the extract concentration, total phenolic compounds and flavonoid compounds increase (p <0.05). Due to the better performance, the enzymatic extracted chlorophyll was used to evaluate the antioxidant properties, inhibition of free radicals DPPH and ATBS and ferric reducing antioxidant power and the result shows the higher chlorophyll concentration, the higher antioxidant properties. Due to the appropriate antioxidant effect of alfalfa chlorophyll extracted by enzymatic method, its application in food, pharmaceutical and health industries can be evaluated and industrial scale extraction systems can be designed using this technique.

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The local strain of B. thuringiensis var. tenebrionis-BN2 (Btt-BN2) was used to control the alfalfa weevil. Experimental design using Response Surface Methodology (RSM) was applied for the optimization of the culture medium and fermentation parameters in order to achieve a high level of Colony-Forming Unit (CFU) (spore mL⁻¹). The parameters including the amount of carbon and nitrogen sources at three levels were investigated with CFU (spore mL⁻¹) response for two sets of experiments. The obtained results showed that the maximum CFU (spore mL⁻¹) for the minimum concentration of oat (2 g L^-1), the minimum concentration of corn steep liquor (10 g L^-1), and the maximum concentration of sugarcane molasses (10 g L^-1) were equivalent to 1.4×10¹³ spore mL^-1. Optimum fermentation parameters to obtain the highest value of CFU (spore mL⁻¹) were determined as a maximum level of pH of eight and a medium level of temperature (28⁰C). Amazingly, optimum conditions enhanced the CFU value to 8.06× 10¹³ spore mL^-1, which is very significant in the Btt research. Finally, the bioassay analysis of Btt in a single system and binary system (combination of two insecticides; Matrine^® and Abamectin^®) at different concentrations illustrated 83% of mortality efficiency (3 ppm of Btt and 0.5 ppm Matrine) on the 3^rd day of treatment and 100% efficiency almost for all combination of Btt with Matrine and/or Abamectin after the 7^th day. The bioassay results showed promising environmentally friendly mortality efficiency compared to the current chemical treatments.