Academic literature on the topic 'Other agricultural, veterinary and food sciences not elsewhere classified'

Unmanned Aerial Vehicles UAVs or Drones have made great progress in the field of aerial surveys to study vegetation and farmland. The research focuses on developing smart systems for managing agricultural fields, thus facilitating decision-making, increasing agricultural productivity, improving profitability and protecting the environment. The paper highlights the ability of drones to distinguish agricultural land intended for cultivation and classified as deserted or cultivated or in the germination stage. For the first time in the Nineveh governorate, a Phantom 4 DJI UAV images were used, in addition to using the spatialized Pix4Dfielde program to process these images. Four types of the standard agricultural indices that rely on the visible spectrum have been used (Visible Atmospherically Resistant Index (VARI), Triangular Greenness Index (TGI), Synthetic Normalized Differences Vegetation Index (S-NDVI) and Visible Difference Vegetation Index (VDVI)) to test UAVs images and to categorize different types of agricultural land. The results showed that when using the S-NDVI and VDVI indicators, the values 0.16 and 0.14 appeared respectively in certain areas, which indicates the presence and integrity of vegetation cover, unlike other regions, whose indicators showed 0.010 and -0.004, respectively, which indicate that the plant has a bad condition or its absence at all. All results finding in this research reflect and confirm the validity of using UAVs images for agricultural field management and development.

BackgroundThe diet of the koala (Phascolarctos cinereus) is comprised almost exclusively of foliage from the genusEucalyptus(family Myrtaceae).Eucalyptusproduces a wide variety of potentially toxic plant secondary metabolites which have evolved as chemical defences against herbivory. The koala is classified as an obligate dietary specialist, and although dietary specialisation is rare in mammalian herbivores, it has been found elsewhere to promote a highly-conserved but low-diversity gut microbiome. The gut microbes of dietary specialists have been found sometimes to enhance tolerance of dietary PSMs, facilitating competition-free access to food. Although the koala and its gut microbes have evolved together to utilise a low nutrient, potentially toxic diet, their gut microbiome has not previously been assessed in conjunction with diet quality. Thus, linking the two may provide new insights in to the ability of the koala to extract nutrients and detoxify their potentially toxic diet.MethodThe 16S rRNA gene was used to characterise the composition and diversity of faecal bacterial communities from a wild koala population (n = 32) comprising individuals that predominately eat either one of two different food species, one the strongly preferred and relatively nutritious speciesEucalyptus viminalis, the other comprising the less preferred and less digestible speciesEucalyptus obliqua.ResultsAlpha diversity indices indicated consistently and significantly lower diversity and richness in koalas eatingE. viminalis. Assessment of beta diversity using both weighted and unweighted UniFrac matrices indicated that diet was a strong driver of both microbial community structure, and of microbial presence/absence across the combined koala population and when assessed independently. Further, principal coordinates analysis based on both the weighted and unweighted UniFrac matrices for the combined and separated populations, also revealed a separation linked to diet. During our analysis of the OTU tables we also detected a strong association between microbial community composition and host diet. We found that the phyla Bacteroidetes and Firmicutes were co-dominant in all faecal microbiomes, with Cyanobacteria also co-dominant in some individuals; however, theE. viminalisdiet produced communities dominated by the generaParabacteroidesand/orBacteroides, whereas theE. obliqua-associated diets were dominated by unidentified genera from the family Ruminococcaceae.DiscussionWe show that diet differences, even those caused by differential consumption of the foliage of two species from the same plant genus, can profoundly affect the gut microbiome of a specialist folivorous mammal, even amongst individuals in the same population. We identify key microbiota associated with each diet type and predict functions within the microbial community based on 80 previously identifiedParabacteroidesand Ruminococcaceae genomes.

From 2017 to 2019, 51 A. rabiei isolates were isolated from 259 chickpea fields across IKR. On different media, 32 isolates showed significant differences in morphological characteristics. The isolates were divided into six groups based on colony color, five groups on mycelium color and three groups on pycnidia color. CSMDA was the best media for mycelial growth. AS-28 significantly surpassed all other isolates in colony diameter despite media type. A. rabiei growth varied between 15-35°C, the maximum growth occured at 25°C and ceased at 35°C. The mean conidia and pycnedia dimensions in isolates AS-19 and AS-9 ranged from 20.0*7.5μm and 70.8*47.9μm to 21.8*9.0μm and 140.7*93.6μm in AS-11 and AS-18 respectively. The isolates were classified into four groups and 15 races based on pathogenicity and virulence. Race 1 exhibited high aggressiveness and virulence against all differentials, whereas the other races explored variable virulence spectrum. Sulaimani had the greatest A. rabiei diversity, with nine different races accounting for 60% of population, followed by Erbil with five races (33%). Halabja and Garmian each contribute three and two races, accounting for 20% and 12% of the total. Races 4 and 5 were the most populous and widely spread in IKR. The ITS region was amplified to a541bp band in all A. rabiei isolates. The length of the nucleotide sequences ranged from 481 to 541bp. The ITS sequences of all the isolates were registered at the NCBI Gen Bank under different accession numbers. The phylogenetic tree clearly shows that all the isolates are grouped in one cluster and have a high degree of similarity.

Environments containing significant concentration of NaCl such as salt lakes harbor extremophiles microorganisms which have a great biotechnology interest. To explore the diversity of Bacteria in Chott Tinsilt (Algeria), an isolation program was performed. Water samples were collected from the saltern during the pre-salt harvesting phase. This Chott is high in salt (22.47% (w/v). Seven halophiles Bacteria were selected for further characterization. The isolated strains were able to grow optimally in media with 10–25% (w/v) total salts. Molecular identification of the isolates was performed by sequencing the 16S rRNA gene. It showed that these cultured isolates included members belonging to the Halomonas, Staphylococcus, Salinivibrio, Planococcus and Halobacillus genera with less than 98% of similarity with their closest phylogenetic relative. The halophilic bacterial isolates were also characterized for the production of biosurfactant and industrially important enzymes. Most isolates produced hydrolases and biosurfactants at high salt concentration. In fact, this is the first report on bacterial strains (A4 and B4) which were a good biosurfactant and coagulase producer at 20% and 25% ((w/v)) NaCl. In addition, the biosurfactant produced by the strain B4 at high salinity (25%) was also stable at high temperature (30-100°C) and high alkalinity (pH 11).Key word: Salt Lake, Bacteria, biosurfactant, Chott, halophiles, hydrolases, 16S rRNAINTRODUCTIONSaline lakes cover approximately 10% of the Earth’s surface area. The microbial populations of many hypersaline environments have already been studied in different geographical regions such as Great Salt Lake (USA), Dead Sea (Israel), Wadi Natrun Lake (Egypt), Lake Magadi (Kenya), Soda Lake (Antarctica) and Big Soda Lake and Mono Lake (California). Hypersaline regions differ from each other in terms of geographical location, salt concentration and chemical composition, which determine the nature of inhabitant microorganisms (Gupta et al., 2015). Then low taxonomic diversity is common to all these saline environments (Oren et al., 1993). Halophiles are found in nearly all major microbial clades, including prokaryotic (Bacteria and Archaea) and eukaryotic forms (DasSarma and Arora, 2001). They are classified as slight halophiles when they grow optimally at 0.2–0.85 M (2–5%) NaCl, as moderate halophiles when they grow at 0.85–3.4 M (5–20%) NaCl, and as extreme halophiles when they grow at 3.4–5.1 M (20–30%) NaCl. Hyper saline environments are inhabited by extremely halophilic and halotolerant microorganisms such as Halobacillus sp, Halobacterium sp., Haloarcula sp., Salinibacter ruber , Haloferax sp and Bacillus spp. (Solomon and Viswalingam, 2013). There is a tremendous demand for halophilic bacteria due to their biotechnological importance as sources of halophilic enzymes. Enzymes derived from halophiles are endowed with unique structural features and catalytic power to sustain the metabolic and physiological processes under high salt conditions. Some of these enzymes have been reported to be active and stable under more than one extreme condition (Karan and Khare, 2010). Applications are being considered in a range of industries such as food processing, washing, biosynthetic processes and environmental bioremediation. Halophilic proteases are widely used in the detergent and food industries (DasSarma and Arora, 2001). However, esterases and lipases have also been useful in laundry detergents for the removal of oil stains and are widely used as biocatalysts because of their ability to produce pure compounds. Likewise, amylases are used industrially in the first step of the production of high fructose corn syrup (hydrolysis of corn starch). They are also used in the textile industry in the de-sizing process and added to laundry detergents. Furthermore, for the environmental applications, the use of halophiles for bioremediation and biodegradation of various materials from industrial effluents to soil contaminants and accidental spills are being widely explored. In addition to enzymes, halophilic / halotolerants microorganisms living in saline environments, offer another potential applications in various fields of biotechnology like the production of biosurfactant. Biosurfactants are amphiphilic compounds synthesized from plants and microorganisms. They reduce surface tension and interfacial tension between individual molecules at the surface and interface respectively (Akbari et al., 2018). Comparing to the chemical surfactant, biosurfactant are promising alternative molecules due to their low toxicity, high biodegradability, environmental capability, mild production conditions, lower critical micelle concentration, higher selectivity, availability of resources and ability to function in wide ranges of pH, temperature and salinity (Rocha et al., 1992). They are used in various industries which include pharmaceuticals, petroleum, food, detergents, cosmetics, paints, paper products and water treatment (Akbari et al., 2018). The search for biosurfactants in extremophiles is particularly promising since these biomolecules can adapt and be stable in the harsh environments in which they are to be applied in biotechnology.OBJECTIVESEastern Algeria features numerous ecosystems including hypersaline environments, which are an important source of salt for food. The microbial diversity in Chott Tinsilt, a shallow Salt Lake with more than 200g/L salt concentration and a superficies of 2.154 Ha, has never yet been studied. The purpose of this research was to chemically analyse water samples collected from the Chott, isolate novel extremely or moderate halophilic Bacteria, and examine their phenotypic and phylogenetic characteristics with a view to screening for biosurfactants and enzymes of industrial interest.MATERIALS AND METHODSStudy area: The area is at 5 km of the Commune of Souk-Naâmane and 17 km in the South of the town of Aïn-Melila. This area skirts the trunk road 3 serving Constantine and Batna and the railway Constantine-Biskra. It is part the administrative jurisdiction of the Wilaya of Oum El Bouaghi. The Chott belongs to the wetlands of the High Plains of Constantine with a depth varying rather regularly without never exceeding 0.5 meter. Its length extends on 4 km with a width of 2.5 km (figure 1).Water samples and physico-chemical analysis: In February 2013, water samples were collected from various places at the Chott Tinsilt using Global Positioning System (GPS) coordinates of 35°53’14” N lat. and 06°28’44”E long. Samples were collected randomly in sterile polythene bags and transported immediately to the laboratory for isolation of halophilic microorganisms. All samples were treated within 24 h after collection. Temperature, pH and salinity were measured in situ using a multi-parameter probe (Hanna Instruments, Smithfield, RI, USA). The analytical methods used in this study to measure ions concentration (Ca2+, Mg2+, Fe2+, Na+, K+, Cl−, HCO3−, SO42−) were based on 4500-S-2 F standard methods described elsewhere (Association et al., 1920).Isolation of halophilic bacteria from water sample: The media (M1) used in the present study contain (g/L): 2.0 g of KCl, 100.0/200.0 g of NaCl, 1.0 g of MgSO4.7HO2, 3.0 g of Sodium Citrate, 0.36 g of MnCl2, 10.0 g of yeast extract and 15.0 g agar. The pH was adjusted to 8.0. Different dilutions of water samples were added to the above medium and incubated at 30°C during 2–7 days or more depending on growth. Appearance and growth of halophilic bacteria were monitored regularly. The growth was diluted 10 times and plated on complete medium agar (g/L): glucose 10.0; peptone 5.0; yeast extract 5.0; KH2PO4 5.0; agar 30.0; and NaCl 100.0/200.0. Resultant colonies were purified by repeated streaking on complete media agar. The pure cultures were preserved in 20% glycerol vials and stored at −80°C for long-term preservation.Biochemical characterisation of halophilic bacterial isolates: Bacterial isolates were studied for Gram’s reaction, cell morphology and pigmentation. Enzymatic assays (catalase, oxidase, nitrate reductase and urease), and assays for fermentation of lactose and mannitol were done as described by Smibert (1994).Optimization of growth conditions: Temperature, pH, and salt concentration were optimized for the growth of halophilic bacterial isolates. These growth parameters were studied quantitatively by growing the bacterial isolates in M1 medium with shaking at 200 rpm and measuring the cell density at 600 nm after 8 days of incubation. To study the effect of NaCl on the growth, bacterial isolates were inoculated on M1 medium supplemented with different concentration of NaCl: 1%-35% (w/v). The effect of pH on the growth of halophilic bacterial strains was studied by inoculating isolates on above described growth media containing NaCl and adjusted to acidic pH of 5 and 6 by using 1N HCl and alkaline pH of 8, 9, 10, 11 and 12 using 5N NaOH. The effect of temperature was studied by culturing the bacterial isolates in M1 medium at different temperatures of incubation (4°C–55°C).Screening of halophilic bacteria for hydrolytic enzymes: Hydrolase producing bacteria among the isolates were screened by plate assay on starch, tributyrin, gelatin and DNA agar plates respectively for amylase, lipase, protease and DNAse activities. Amylolytic activity of the cultures was screened on starch nutrient agar plates containing g/L: starch 10.0; peptone 5.0; yeast extract 3.0; agar 30.0; NaCl 100.0/250.0. The pH was 7.0. After incubation at 30 ºC for 7 days, the zone of clearance was determined by flooding the plates with iodine solution. The potential amylase producers were selected based on ratio of zone of clearance diameter to colony diameter. Lipase activity of the cultures was screened on tributyrin nutrient agar plates containing 1% (v/v) of tributyrin. Isolates that showed clear zones of tributyrin hydrolysis were identified as lipase producing bacteria. Proteolytic activity of the isolates was similarly screened on gelatin nutrient agar plates containing 10.0 g/L of gelatin. The isolates showing zones of gelatin clearance upon treatment with acidic mercuric chloride were selected and designated as protease producing bacteria. The presence of DNAse activity on plates was determined on DNAse test agar (BBL) containing 10%-25% (w/v) total salt. After incubation for 7days, the plates were flooded with 1N HCl solution. Clear halos around the colonies indicated DNAse activity (Jeffries et al., 1957).Milk clotting activity (coagulase activity) of the isolates was also determined following the procedure described (Berridge, 1952). Skim milk powder was reconstituted in 10 mM aqueous CaCl2 (pH 6.5) to a final concentration of 0.12 kg/L. Enzyme extracts were added at a rate of 0.1 mL per mL of milk. The coagulation point was determined by manual rotating of the test tube periodically, at short time intervals, and checking for visible clot formation.Screening of halophilic bacteria for biosurfactant production. Oil spread Assay: The Petridis base was filled with 50 mL of distilled water. On the water surface, 20μL of diesel and 10μl of culture were added respectively. The culture was introduced at different spots on the diesel, which is coated on the water surface. The occurrence of a clear zone was an indicator of positive result (Morikawa et al., 2000). The diameter of the oil expelling circles was measured by slide caliber (with a degree of accuracy of 0.02 mm).Surface tension and emulsification index (E24): Isolates were cultivated at 30 °C for 7 days on the enrichment medium containing 10-25% NaCl and diesel oil as the sole carbon source. The medium was centrifuged (7000 rpm for 20 min) and the surface tension of the cell-free culture broth was measured with a TS90000 surface tensiometer (Nima, Coventry, England) as a qualitative indicator of biosurfactant production. The culture broth was collected with a Pasteur pipette to remove the non-emulsified hydrocarbons. The emulsifying capacity was evaluated by an emulsification index (E24). The E24 of culture samples was determined by adding 2 mL of diesel oil to the same amount of culture, mixed for 2 min with a vortex, and allowed to stand for 24 h. E24 index is defined as the percentage of height of emulsified layer (mm) divided by the total height of the liquid column (mm).Biosurfactant stability studies : After growth on diesel oil as sole source of carbone, cultures supernatant obtained after centrifugation at 6,000 rpm for 15 min were considered as the source of crude biosurfactant. Its stability was determined by subjecting the culture supernatant to various temperature ranges (30, 40, 50, 60, 70, 80 and 100 °C) for 30 min then cooled to room temperature. Similarly, the effect of different pH (2–11) on the activity of the biosurfactant was tested. The activity of the biosurfactant was investigated by measuring the emulsification index (El-Sersy, 2012).Molecular identification of potential strains. DNA extraction and PCR amplification of 16S rDNA: Total cellular DNA was extracted from strains and purified as described by Sambrook et al. (1989). DNA was purified using Geneclean® Turbo (Q-BIO gene, Carlsbad, CA, USA) before use as a template in polymerase chain reaction (PCR) amplification. For the 16S rDNA gene sequence, the purified DNA was amplified using a universal primer set, forward primer (27f; 5′-AGA GTT TGA TCM TGG CTC AG) and a reverse primer (1492r; 5′-TAC GGY TAC CTT GTT ACG ACT T) (Lane, 1991). Agarose gel electrophoresis confirmed the amplification product as a 1400-bp DNA fragment.16S rDNA sequencing and Phylogenic analysis: Amplicons generated using primer pair 27f-1492r was sequenced using an automatic sequencer system at Macrogene Company (Seoul, Korea). The sequences were compared with those of the NCBI BLAST GenBank nucleotide sequence databases. Phylogenetic trees were constructed by the neighbor-joining method using MEGA version 5.05 software (Tamura et al., 2011). Bootstrap resembling analysis for 1,000 replicates was performed to estimate the confidence of tree topologies.Nucleotide sequence accession numbers: The nucleotide sequences reported in this work have been deposited in the EMBL Nucleotide Sequence Database. The accession numbers are represented in table 5.Statistics: All experiments were conducted in triplicates. Results were evaluated for statistical significance using ANOVA.RESULTSPhysico-chemical parameters of the collected water samples: The physicochemical properties of the collected water samples are reported in table 1. At the time of sampling, the temperature was 10.6°C and pH 7.89. The salinity of the sample, as determined in situ, was 224.70 g/L (22,47% (w/v)). Chemical analysis of water sample indicated that Na +and Cl- were the most abundant ions (table 1). SO4-2 and Mg+2 was present in much smaller amounts compared to Na +and Cl- concentration. Low levels of calcium, potassium and bicarbonate were also detected, often at less than 1 g/L.Characterization of isolates. Morphological and biochemical characteristic feature of halophilic bacterial isolates: Among 52 strains isolated from water of Chott Tinsilt, seven distinct bacteria (A1, A2, A3, A4, B1, B4 and B5) were chosen for further characterization (table 2). The colour of the isolates varied from beige, pale yellow, yellowish and orange. The bacterial isolates A1, A2, A4, B1 and B5 were rod shaped and gram negative (except B5), whereas A3 and B4 were cocci and gram positive. All strains were oxidase and catalase positive except for B1. Nitrate reductase and urease activities were observed in all the bacterial isolates, except B4. All the bacterial isolates were negative for H2S formation. B5 was the only strain positive for mannitol fermentation (table 2).We isolated halophilic bacteria on growth medium with NaCl supplementation at pH 7 and temperature of 30°C. We studied the effect of NaCl, temperature and pH on the growth of bacterial isolates. All the isolates exhibited growth only in the presence of NaCl indicating that these strains are halophilic. The optimum growth of isolates A3 and B1 was observed in the presence of 10% NaCl, whereas it was 15% NaCl for A1, A2 and B5. A4 and B4 showed optimum growth in the presence of 20% and 25% NaCl respectively. A4, B4 and B5 strains can tolerate up to 35% NaCl.The isolate B1 showed growth in medium supplemented with 10% NaCl and pH range of 7–10. The optimum pH for the growth B1 was 9 and they did not show any detectable growth at or below pH 6 (table 2), which indicates the alkaliphilic nature of B1 isolate. The bacterial isolates A1, A2 and A4 exhibited growth in the range of pH 6–10, while A3 and B4 did not show any growth at pH greater than 8. The optimum pH for growth of all strains (except B1) was pH 7.0 (table 2). These results indicate that A1, A2, A3, A4, B4 and B5 are neutrophilic in nature. All the bacterial isolates exhibited optimal growth at 30°C and no detectable growth at 55°C. Also, detectable growth of isolates A1, A2 and A4 was observed at 4°C. However, none of the bacterial strains could grow below 4°C and above 50°C (table 2).Screening of the halophilic enzymes: To characterize the diversity of halophiles able to produce hydrolytic enzymes among the population of microorganisms inhabiting the hypersaline habitats of East Algeria (Chott Tinsilt), a screening was performed. As described in Materials and Methods, samples were plated on solid media containing 10%-25% (w/v) of total salts and different substrates for the detection of amylase, protease, lipase and DNAse activities. However, coagulase activity was determined in liquid medium using milk as substrate (figure 3). Distributions of hydrolytic activity among the isolates are summarized in table 4.From the seven bacterial isolates, four strains A1, A2, A4 and B5 showed combined hydrolytic activities. They were positive for gelatinase, lipase and coagulase. A3 strain showed gelatinase and lipase activities. DNAse activities were detected with A1, A4, B1 and B5 isolates. B4 presented lipase and coagulase activity. Surprisingly, no amylase activity was detected among all the isolates.Screening for biosurfactant producing isolates: Oil spread assay: The results showed that all the strains could produce notable (>4 cm diameter) oil expelling circles (ranging from 4.11 cm to 4.67 cm). The average diameter for strain B5 was 4.67 cm, significantly (P < 0.05) higher than for the other strains.Surface tension and emulsification index (E24): The assimilation of hydrocarbons as the sole sources of carbon by the isolate strains led to the production of biosurfactants indicated by the emulsification index and the lowering of the surface tension of cell-free supernatant. Based on rapid growth on media containing diesel oil as sole carbon source, the seven isolates were tested for biosurfactant production and emulsification activity. The obtained values of the surface tension measurements as well as the emulsification index (E24) are shown in table 3. The highest reduction of surface tension was achieved with B5 and A3 isolates with values of 25.3 mN m−1 and 28.1 mN m−1 respectively. The emulsifying capacity evaluated by the E24 emulsification index was highest in the culture of isolate B4 (78%), B5 (77%) and A3 (76%) as shown in table 3 and figure 2. These emulsions were stable even after 4 months. The bacteria with emulsification indices higher than 50 % and/or reduction in the surface tension (under 30 mN/m) have been defined as potential biosurfactant producers. Based on surface tension and the E24 index results, isolates B5, B4, A3 and A4 are the best candidates for biosurfactant production. It is important to note that, strains B4 and A4 produce biosurfactant in medium containing respectively 25% and 20% (w/v) NaCl.Stability of biosurfactant activities: The applicability of biosurfactants in several biotechnological fields depends on their stability at different environmental conditions (temperatures, pH and NaCl). For this study, the strain B4 appear very interesting (It can produce biosurfactant at 25 % NaCl) and was choosen for futher analysis for biosurfactant stability. The effects of temperature and pH on the biosurfactant production by the strain B4 are shown in figure 4.biosurfactant in medium containing respectively 25% and 20% (w/v) NaCl.Stability of biosurfactant activities: The applicability of biosurfactants in several biotechnological fields depends on their stability at different environmental conditions (temperatures, pH and NaCl). For this study, the strain B4 appear very interesting (It can produce biosurfactant at 25 % NaCl) and was chosen for further analysis for biosurfactant stability. The effects of temperature and pH on the biosurfactant production by the strain B4 are shown in figure 4. The biosurfactant produced by this strain was shown to be thermostable giving an E-24 Index value greater than 78% (figure 4A). Heating of the biosurfactant to 100 °C caused no significant effect on the biosurfactant performance. Therefore, the surface activity of the crude biosurfactant supernatant remained relatively stable to pH changes between pH 6 and 11. At pH 11, the value of E24 showed almost 76% activity, whereas below pH 6 the activity was decreased up to 40% (figure 4A). The decreases of the emulsification activity by decreasing the pH value from basic to an acidic region; may be due to partial precipitation of the biosurfactant. This result indicated that biosurfactant produced by strain B4 show higher stability at alkaline than in acidic conditions.Molecular identification and phylogenies of potential isolates: To identify halophilic bacterial isolates, the 16S rDNA gene was amplified using gene-specific primers. A PCR product of ≈ 1.3 kb was detected in all the seven isolates. The 16S rDNA amplicons of each bacterial isolate was sequenced on both strands using 27F and 1492R primers. The complete nucleotide sequence of 1336,1374, 1377,1313, 1305,1308 and 1273 bp sequences were obtained from A1, A2, A3, A4, B1, B4 and B5 isolates respectively, and subjected to BLAST analysis. The 16S rDNA sequence analysis showed that the isolated strains belong to the genera Halomonas, Staphylococcus, Salinivibrio, Planococcus and Halobacillus as shown in table 5. The halophilic isolates A2 and A4 showed 97% similarity with the Halomonas variabilis strain GSP3 (accession no. AY505527) and the Halomonas sp. M59 (accession no. AM229319), respectively. As for A1, it showed 96% similarity with the Halomonas venusta strain GSP24 (accession no. AY553074). B1 and B4 showed for their part 96% similarity with the Salinivibrio costicola subsp. alcaliphilus strain 18AG DSM4743 (accession no. NR_042255) and the Planococcus citreus (accession no. JX122551), respectively. The bacterial isolate B5 showed 98% sequence similarity with the Halobacillus trueperi (accession no. HG931926), As for A3, it showed only 95% similarity with the Staphylococcus arlettae (accession no. KR047785). The 16S rDNA nucleotide sequences of all the seven halophilic bacterial strains have been submitted to the NCBI GenBank database under the accession number presented in table 5. The phylogenetic association of the isolates is shown in figure 5.DICUSSIONThe physicochemical properties of the collected water samples indicated that this water was relatively neutral (pH 7.89) similar to the Dead Sea and the Great Salt Lake (USA) and in contrast to the more basic lakes such as Lake Wadi Natrun (Egypt) (pH 11) and El Golea Salt Lake (Algeria) (pH 9). The salinity of the sample was 224.70 g/L (22,47% (w/v). This range of salinity (20-30%) for Chott Tinsilt is comparable to a number of well characterized hypersaline ecosystems including both natural and man-made habitats, such as the Great Salt Lake (USA) and solar salterns of Puerto Rico. Thus, Chott Tinsilt is a hypersaline environment, i.e. environments with salt concentrations well above that of seawater. Chemical analysis of water sample indicated that Na +and Cl- were the most abundant ions, as in most hypersaline ecosystems (with some exceptions such as the Dead Sea). These chemical water characteristics were consistent with the previously reported data in other hypersaline ecosystems (DasSarma and Arora, 2001; Oren, 2002; Hacěne et al., 2004). Among 52 strains isolated from this Chott, seven distinct bacteria (A1, A2, A3, A4, B1, B4 and B5) were chosen for phenotypique, genotypique and phylogenetique characterization.The 16S rDNA sequence analysis showed that the isolated strains belong to the genera Halomonas, Staphylococcus, Salinivibrio, Planococcus and Halobacillus. Genera obtained in the present study are commonly occurring in various saline habitats across the globe. Staphylococci have the ability to grow in a wide range of salt concentrations (Graham and Wilkinson, 1992; Morikawa et al., 2009; Roohi et al., 2014). For example, in Pakistan, Staphylococcus strains were isolated from various salt samples during the study conducted by Roohi et al. (2014) and these results agreed with previous reports. Halomonas, halophilic and/or halotolerant Gram-negative bacteria are typically found in saline environments (Kim et al., 2013). The presence of Planococcus and Halobacillus has been reported in studies about hypersaline lakes; like La Sal del Rey (USA) (Phillips et al., 2012) and Great Salt Lake (Spring et al., 1996), respectively. The Salinivibrio costicola was a representative model for studies on osmoregulatory and other physiological mechanisms of moderately halophilic bacteria (Oren, 2006).However, it is interesting to note that all strains shared less than 98.7% identity (the usual species cut-off proposed by Yarza et al. (2014) with their closest phylogenetic relative, suggesting that they could be considered as new species. Phenotypic, genetic and phylogenetic analyses have been suggested for the complete identification of these strains. Theses bacterial strains were tested for the production of industrially important enzymes (Amylase, protease, lipase, DNAse and coagulase). These isolates are good candidates as sources of novel enzymes with biotechnological potential as they can be used in different industrial processes at high salt concentration (up to 25% NaCl for B4). Prominent amylase, lipase, protease and DNAase activities have been reported from different hypersaline environments across the globe; e.g., Spain (Sánchez‐Porro et al., 2003), Iran (Rohban et al., 2009), Tunisia (Baati et al., 2010) and India (Gupta et al., 2016). However, to the best of our knowledge, the coagulase activity has never been detected in extreme halophilic bacteria. Isolation and characterization of crude enzymes (especially coagulase) to investigate their properties and stability are in progress.The finding of novel enzymes with optimal activities at various ranges of salt concentrations is of great importance. Besides being intrinsically stable and active at high salt concentrations, halophilic and halotolerant enzymes offer great opportunities in biotechnological applications, such as environmental bioremediation (marine, oilfiel) and food processing. The bacterial isolates were also characterized for production of biosurfactants by oil-spread assay, measurement of surface tension and emulsification index (E24). There are few reports on biosurfactant producers in hypersaline environments and in recent years, there has been a greater increase in interest and importance in halophilic bacteria for biomolecules (Donio et al., 2013; Sarafin et al., 2014). Halophiles, which have a unique lipid composition, may have an important role to play as surface-active agents. The archae bacterial ether-linked phytanyl membrane lipid of the extremely halophilic bacteria has been shown to have surfactant properties (Post and Collins, 1982). Yakimov et al. (1995) reported the production of biosurfactant by a halotolerant Bacillus licheniformis strain BAS 50 which was able to produce a lipopeptide surfactant when cultured at salinities up to 13% NaCl. From solar salt, Halomonas sp. BS4 and Kocuria marina BS-15 were found to be able to produce biosurfactant when cultured at salinities of 8% and 10% NaCl respectively (Donio et al., 2013; Sarafin et al., 2014). In the present work, strains B4 and A4 produce biosurfactant in medium containing respectively 25% and 20% NaCl. To our knowledge, this is the first report on biosurfactant production by bacteria under such salt concentration. Biosurfactants have a wide variety of industrial and environmental applications (Akbari et al., 2018) but their applicability depends on their stability at different environmental conditions. The strain B4 which can produce biosurfactant at 25% NaCl showed good stability in alkaline pH and at a temperature range of 30°C-100°C. Due to the enormous utilization of biosurfactant in detergent manufacture the choice of alkaline biosurfactant is researched (Elazzazy et al., 2015). On the other hand, the interesting finding was the thermostability of the produced biosurfactant even after heat treatment (100°C for 30 min) which suggests the use of this biosurfactant in industries where heating is of a paramount importance (Khopade et al., 2012). To date, more attention has been focused on biosurfactant producing bacteria under extreme conditions for industrial and commercial usefulness. In fact, the biosurfactant produce by strain B4 have promising usefulness in pharmaceutical, cosmetics and food industries and for bioremediation in marine environment and Microbial enhanced oil recovery (MEOR) where the salinity, temperature and pH are high.CONCLUSIONThis is the first study on the culturable halophilic bacteria community inhabiting Chott Tinsilt in Eastern Algeria. Different genera of halotolerant bacteria with different phylogeneticaly characteristics have been isolated from this Chott. Culturing of bacteria and their molecular analysis provides an opportunity to have a wide range of cultured microorganisms from extreme habitats like hypersaline environments. Enzymes produced by halophilic bacteria show interesting properties like their ability to remain functional in extreme conditions, such as high temperatures, wide range of pH, and high salt concentrations. 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Drought is a wide-spread problem seriously influencing wheat (Triticum aestivum L.) production , but development of tolerant genotypes is hampered by the lack of effective selection criteria. The objective of research study was to evaluate the efficiency of several selection indices to identify drought tolerant genotypes under drought stress conditions. Twenty seven bread wheat genotypes differing in yield performance were grown under drought stress, and normal irrigation during 2014-2015 growing season, were evaluated in split plot design with three replications Significant and high positive correlation was found between grain yield in the stress condition (Ys) and (Yp) with indices STI, GMP, YI, and Significant negative correlation was found between Ys with SSI, SI, SSPI, TOL indices. Principal component analysis (PCA) based on the Spearman’s correlation matrix, indicated that first PCA (80.6%) and second PCA (18.1%) accounted for 98.7% of variations among the indices. The results of PCA revealed that the screening methods were significantly inter-correlated with each other indicating that several of the statistics probably measure similar aspects of drought tolerance. Cluster analysis classified the cultivars into four groups according to drought tolerance. The results showed that MP, GMP and STI were more effective in identifying high yielding genotypes in both drought-stressed and irrigated conditions, identifying G1 and G10 as more tolerant and G25 and G26 as more sensitive genotype to drought stress.

Poisoning of cattle by plants of the Pimelea species incurs heavy losses to individual producers in regions of South West Queensland, North Western New South Wales, South Australia, and the Northern Territory. Due to the vast

areas of affected land, control of the plant is perceived as non -viable. Therefore, the aim of this study has been to contribute to the development of methods to reduce the losses caused by the condition from an animal health

perspective.

Immunisation with conjugates of a purified mixture of Pimelea toxins and ovalbumin with varying inclusion ratios has resulted in the production of bovine serum IgG antibodies, which attenuate the effect of purified Pimelea

toxins on bovine pulmonary venule preparations in vitro. Cattle immunised with a vaccine batched from two conjugates from experimental studies have been shown to develop toxin specific antibody responses while affected and unaffected by poisoning at the time of immunisation. Experiments to evaluate the protective attributes of the vaccine under field conditions have produced consistent, yet non -significant results regarding liveweight and condition

scores of vaccinated animals. Therefore, no immediate conclusions can be drawn regarding the protective properties of the experimental vaccine without futrther investigation.

Investigation of a structure -activity relationship of the Pimelea toxins has resulted in new knowledge regarding key functional requirements for binding to and activation of protein kinase C (PKC). A structural variant of simplexin

was found not to activate PKC in vitro, therefore the conversion of simplexin to this compound in the rumen of cattle would provide a means of reducing the impact caused by Pimelea poisoning. It was found that hydroxyl groups at the 4- and 5- positions of simplexin were essential for PKC binding. Carbonyl functionality at position 3- of simplexin, although significant for PKC binding, was shown to be more related to the activation properties of the resulting complex. The alteration of hydroxyl functionality at the 20- position of

mezerein (a selected reference compound) was shown not to significantly alter binding, however no conclusion can be drawn regarding the importance of this functionality of simplexin.

Derivatisation of Pimelea toxin for assay by gas chromatography (GC) and high performance liquid chromatography (HPLC) was found not to be feasible.

The poultry industry has rapidly grown in the last few years with a focus in improving growth and productivity of broiler chickens, with performance assessed on measures such as feed conversion ratio, body weight gain and good immunity. The incorporation of antibiotics and feed additives in poultry diets, have been implemented for years to ensure the maintenance of poultry health with a focus on the control and reduction of zoonotic pathogens. In the last few years, however, key issues surrounding the antimicrobial resistance of antibiotics have urged for alternative supplementations. Nanoparticles (NPs) of silver and other metals have been heavily used in the poultry industry to improve the growth and performance of birds. Whilst successful, metal NPs exhibited higher toxicity at the higher surface to volume ratio, especially with the use of silver. This study proposes the use of NPs of essential metals and natural compounds to safely deliver nutrients, resulting in positive impacts on health and productivity with little to no toxic effects. Selenium is an essential mineral, required for the proper functioning of the immunity and is an important element in the first cell line of defence in the body. The work described in this thesis explores the ability of selenium NPs to improve the health and growth of broiler chickens by modulating their gut microbiome and metabolome, without the toxic effects observed with silver. Selenium NPs were synthesised using a simple chemical reduction method and a full characterisation was performed, assessing the physicochemical properties of the NP. Selenium NPs were then compared in an animal trial against two commonly used selenium additives in the poultry industry, sodium selenite (inorganic selenium) and selenomethionine (organic selenium). The performance of the birds was assessed based on body weight gain, the gut microbial composition and metabolite production. The toxicity of NPs was further investigated by quantifying selenium concentration in various tissues, along with a detailed histopathological assessment. Results show selenium NPs completely altered the gut microbial ecology at high concentration, with a strong correlation observed between Faecalibacterium prausnitzii abundance and increasing concentration of selenium NPs. Selenium NPs additionally increased villus height/crypt ratio associated with enhanced absorption in the small intestine and an overall increase of healthy colonic metabolites. Finally, an in vitro study demonstrated the ability of selenium NPs to reduce emerging pathogenic Enterococcus cecorum species. This thesis demonstrates the prospective ability of selenium NPs as alternatives to antibiotics and bulk supplementation, resulting in an improvement of health and performance of broiler chickens in the poultry industry.

Understanding the effect of food insecurity of vulnerable individuals is necessary to develop strategies for improving lives of those individuals. In this study I explore the effect of a low-cost, unobtrusive intervention on food pantry clients’ choice of healthier food items at a local food pantry. A cross-sectional study was conducted at a food pantry in the Midwest U.S. using the randomized controlled trial method. Participants in the intervention group received a nutrition ranking information about the food items in the pantry during their visit. Both the intervention and control groups reported their food selections. Additionally, client demographic information was collected in surveys. Data were collected from October 2018 to January 2019. A total of 615 adults were recruited and randomized for the nutrition ranking intervention (n=300) and control group (n=315). Multiple linear regression models were used to predict the outcomes of the intervention while controlling for demographic characteristics such as age, gender, household size, and education level. There was no significant response to the nutritional ranking intervention as it appears that the intervention was ineffective at changing behavior. Results suggest that future studies are needed to determine a low-cost intervention for food pantry clients during their short time at the food pantry.

Women who operate small-scale farms and sell to small markets in Indiana may encounter certain obstacles and constraints due to self-employment in the traditionally male-dominated field of agriculture. Researchers have recognized the role of sustainable agriculture ideology in attracting women to these niche agricultural markets. Despite increasing opportunities for women in sustainable agriculture, research suggests that traditional gender roles are often maintained, manifesting itself through several obstacles (Pilgeram & Amos, 2015). Female agricultural entrepreneurs encounter obstacles including work-family balance, geographic barriers, access to physical resources, access to financial resources, access to places of information. Previous research indicates that female entrepreneurs have less access to human, social, and financial capital to support their business ventures (Powell & Eddleston, 2013). However, this study explored the feminine perspective and management styles which may serve as beneficial resources.

The purpose of this study was to explore and describe existing obstacles encountered by female entrepreneurs in niche agricultural markets and their methods of building resilience in their business. Quantitative data was collected through an online survey of 62 agricultural entrepreneurs across the state of Indiana. Participants were asked questions pertaining to their business structure, resources, constraints, processes, achievements, and demographics. Several responses to open-ended questions were also collected and analyzed through open, axial coding. Study results include the diversity of the population, value of human capital resources, prioritization of quality products, significance of internal constraints, discrepancies in division of labor and women’s obstacles to access to social networks. A greater understanding of the obstacles encountered by women agricultural entrepreneurs can also provide valuable insight to Land-Grant University Extension, policymakers, and stakeholders in the Indiana agriculture industry.

STEM education is a top priority in the educational development of youth across the United States as the country tries to address the need of having a more well equipped, prepared, and educated workforce. Agriculture, food, and natural resources (AFNR) has the ability to provide a relevant context for engaging students in STEM education through experiential learning. Tragically, both STEM and AFNR struggle to reach and engage more diversified audiences, especially students of color. AFNR education provides an authentic avenue to center STEM engagement around addressing societal grand challenges like food and nutritional security, childhood-obesity, and climate change; issues faced by all communities. The approaches and steps taken to address these AFNR related grand challenges can all be explored through the lens of food systems. Food systems is a concept within AFNR that encompasses the interdisciplinary components of AFNR, STEM, and social sciences that provides a breakdown for the process and system involved in getting food from farm to fork. In an era where youth are more disconnected from understanding where their food comes from, food systems education has the ability to reconnect youth to the root of this issue and the potential to lead them to explore finding solutions to the grand challenges facing their generation. Furthermore, food systems education provides a context to engage youth in authentic learning experiences in nonformal and formal classroom settings around relevant issues with the potential to enhance their interests and concerns around these topics.

The purpose of this study was to explore and describe elementary school students’ interests and concerns about the food system, and their overall engagement in the learning experience after participating in an authentic learning based Virtual Agri+STEM Camp focused on food systems education, AFNR, and STEM activities. The convenience sample for this study was made up of elementary school students between grades 3^rd and 8^th grade (N = 99) who were either in the classroom or participating in an at-home Agri+STEM session. The majority of these students were from rural communities and most of them were African Americans. Quantitative data was collected before and after participation in the Virtual Agri+STEM Camp experience that using the research developed Food System Interest and Food System Concern instrument. Previous AFNR related experiences were also reported by students. The researcher also used an adapted version of the Intrinsic Motivation Inventory (IMI) and STEM Semantics survey to measure student engagement and attitudes after participating in the experience. Descriptive statistics were used to analyze the data, which included means, standard deviations, frequencies, and percentages. To explore the relationships between each of the variables, correlations were also computed.

There were four conclusions for this study. First, students that participated in the Virtual Agri+STEM Camp were motivated and engaged in the learning process while doing the Agri+STEM Camp activities. Second, students that participated in the Virtual Agri+STEM Camp were interested and concerned about the food system before and after participating in the Virtual Agri+STEM Camp. Third, African American student participants reported less previous AFNR experiences, yet they reported more interests and concerns in the food system than Caucasian American participants before and after completing the Virtual Agri+STEM Camp. Lastly, Students that felt more competent, saw the value, and were interested/enjoyed the Agri+STEM experience were more likely to be interested and concerned about the food system. Recommendations for future research and implications for practice and policy were discussed.