Aerobic Spore-Forming Bacteria in the Ultra High Temperature Milk Produced in the North West of Algeria

The occurrence of the spore-forming bacteria in different segments of production lines of Ultra High Temperature （UHT） milk of a commercial plant and its transfer to the final product was studied. The samples were collected from different segments of production lines from reception to packaging sections, over a period of 75 days at a rate of one sampling a week, from plants in North West of Algeria. The total colony count of aerobic spore-forming bacteria was done using Plate Count Agar （PCA） method. The results showed that there was more than 1 ×10^3 CFU/mL at the level of 50%, 25%, 37.5%, 62.5% and 37.5% of the analyzed samples of milk powder, pasteurized milk, UHT milk （before storage）, UHT milk stored at 55℃ for 7 days and UHT milk stored at 30℃ for 14 days, respectively. The mean total colony count of aerobic spore-forming bacteria was considerably lower in the UHT milk than in the other samples, while the higher proportion of aerobic spore-forming bacteria was noticed to be 32.58% and 30.90% in the UHT milk stored at 55℃ and 30℃, respectively. The results obtained by the classic biochemical identification （Bacillus API 20E）, interpreted by software of calculation for microbial identification reflected the presence of especially Bacillus sphaericus. The other species found were： Bacillus cereus, Bacillus subtilis, Bacillus coagulans, Bacillus licheniformis and Bacillus stearothermophilus. The study reveals that the proportion of aerobic spore-forming bacteria in the final product depends on temperature and storage periods.     

Biodegradation of High Concentration of p-Nitrophenol （PNP） by Wastewater Microflora

Many nitrophenols tend to persist in the environment and they may become public health hazards. Among nitrophenols, p-nitrophenol （PNP） is a priority pollutant that has been widely used as pesticide. PNP is a toxic compound that enters the environment during manufacturing and processing of a variety of industrial products. This situation generalized its presence in multiple natural ecosystems： rivers, wastewaters, subterranean waters, soil treated by pesticides and urban atmosphere. This study aims to test the ability of wastewater microflora to degrade high concentration of PNP （500 rag/L） aerobically. An identification of the dominant microorganisms involved in the biodegradation is also carried. The cultures are performed using a minimum medium, where PNP is the sole source of carbon, energy and nitrogen. The kinetic of biodegradation is followed for more than 30 days of incubation at 30 ~C on a shaker （150 tours/min）. The obtained results show that more than 90% of PNP initial concentration is decomposed at the end of incubation. The isolation of microorganisms degrading PNP gave two bacterial colonies with different macroscopic aspects. Sequence analysis of 16S ribosomal DNA indicated that the PNP degrading isolates were closely related to members of the species： Pseudomonas aeruginosa and Bacillus cereus.     

A New Strain of Entomopathogenic Bacillus of the Tea Pest,Eterusia magnifica （Butl.） （Zyganiidae： Lepidoptera） from Darjeeling Foothill Region of India

An entomopathogenic strain of Bacillus sp. was isolated from diseased red slug caterpillars of the leaf-feediug pest of tea, Eterusia magnifica, from the Darjeeling foothill region. Analysis of the bacterimn based on polyphasic approach such as growth phase, biochemical tests, whole body＂ protein, crystal protein profiles along with bioassay （i.e. LC50 and LT50 values） established it as a different strain but close to Bacillus thuringiensis kurstaki （Btk）, the commercial microbial pesticides of lepidopterans. Among biochemical parameters differences were noted between the new strain and Btk in ONPG, lysine decarboxylase, omithin decarboxylase, urease, nitrate reduction, V-P and glucose utilization tests. PAGE analysis of the whole body protein for the new strain recorded a 34 kDa band which was absent in Btk （used as reference）. Crystal protein profile of the newly isolated bacterial strain showed 53 and 49 kDa bands whereas in Btk only 52 kDa band was evident. Although the LC50 values of the new strain and Btk were close, their LT50 values were much different, the new strain showing a lower value than Btk. In light of the above differences and in absence of any report of entomopathogenic bacterial strain of E. magnifica, the isolated strain of Bacillus appeared to be new to science and hence was designated as RS01. The new strain opens up the possibility of its futttre use as microbial pesticide after standardizing its formulation and checking its safety aspects.     

Antimicrobial and Photocatalytic Effect of Silicate and Silicone Hygienic Coatings

Antimicrobial and photocatalytic effect of silicate and silicone paints were tested. Silicate paints are based on water styrene-acrylic dispersion with mineral binder into form of potassium water glass. Silicone paints are based on water acrylic dispersion with silicone resin emulsion. Silicate and silicone paints were formulated with photocatalytic active nanooxides TiO2 and ZnO. Photocatalytic efficiency of coatings was measured as an absorbance change of organic dye Orange II solution at a wavelength of 485 nm. Antimicrobial properties of coatings were tested using agar plate methods. As test microorganisms Pseudomonas aeruginosa, Escherichia coli, Staphylococcus aureus, Penicillium chrysogenum and Aspergillus niger suspension （density 10^6 cells/mL） were used. The inhibition effect of tested coatings and inhibitory zones were evaluated after incubation. Coatings containing nanoparticles of titanium dioxide and zinc oxide showed antimicrobial activity against the test microorganisms. Whereas coating with zinc oxide nanoparticles successfully inhibited the growth of both bacteria and fungi, the photocatalytic nanoparticles of titanium dioxide inhibited bacteria but not fungi （weak inhibition was observed）.     

The Minimal Active Fragment of the CrylAi Toxin is Located Between 36 and 605

The novel crylAi gene that cloned from Bacillus thuringiensis strain SC6H8 encoded a protein exhibiting strong toxicity against Plutella xylostella and Chilo suppressalis in our previous study. Using the available information for the active fragments of other Cry toxins, eight truncated fragments were constructed to identify the minimal active fragment of CrylAi. All truncated fragments were expressed in Escherichia coli strain BL21 （DE3）, and the insecticidal activity against 2ndinstar P. xylostella larvae was assessed using full-length CrylAi as a positive control. The results indicate that the minimal active fragment of the CrylAi toxin against P. xylostella is located between amino acid residues 36^1 and 605^1, which is smaller than the regions previously reported for CrylA. The first two amino acids （34T and 35P） on helix a-1 and whole helix a-2 of domain I and sheet 13-32 of domain III are necessary for CrylAi toxin to keep its toxicity against P. xylostella.     

Acquisition of Insect-Resistant Transgenic Maize Harboring a Truncated crylAh Gene via Agrobacterium-Mediated Transformation

A novel insecticidal gene crylAh was cloned from Bacillus thuringiensis isolate BT8 previously for plant genetic engineering improvement. Truncated active CrylAh toxin has a toxicity level similar to that of the full-length CrylAh toxin. In this study, plant expression vector pMhGM harboring truncated crylAh gene was transformed into maize （Zea mays L.） immature embryos by Agrobacterium tumefaciens-mediated transformation at which maize alcohol dehydrogenase matrix attachment regions （madMARs） were incorporated on both sides of the gene expression cassette to improve gene expression. A total of 23 PCR positive events were obtained with a transformation efficiency of 5% around. Bioassay results showed that events 1-4 and 1-5 exhibited enhanced resistance to the Asian corn borer （Ostriniafurnacalis）. These two events were further confirmed by molecular analysis. Southern blot suggested that a single copy of the crylAh gene was successfully integrated into the maize genome. Western blot and ELISA showed that the foreign gene crylAh was expressed stably at high level in maize and could be inherited stably over generations. The results of a bioassay of T l-T4 transgenic maize plants indicated that the transgenic plants were highly toxic to the Asian corn borer and their resistance could be inherited stably from generation to generation. Thus, events 1-4 and 1-5 are good candidates for the breeding of insect-resistant maize.     

Penetration of a Single Domain of Bacillus thuringiensis Cry1 le-Domain I to a Lipid Membrane In vitro

Domain I of the activated Crystal protein from Bacillus thuringiensis has a seven a-helix bundle structure, which is responsible for membrane channel formation in its insecticidal mechanism. Crylle is toxic to Asian corn borer, Ostrinia furnacalis （Guen6e）, and plays important roles in insect biological control. The domain I from Crylle has been expressed and purified in its normal conformation, as embedded in the full length homologous toxin structure. The membrane insertion ability of this single domain was compared with the full length homologous toxin using a monolayer insertion experiment. The results indicated that the Crylle-domain I had the ability to insert into the lipid monolayer, and this ability is greater than that of the IE648 toxin. However, the state of insertion is not stable and remains for only a short period of time. The Crylle-domain I plays no role in receptor binding as it had a nonspecific binding with the brush border membrane vesicles of the Asian corn borer.     

New Extracellular Bacillus subtilis Lectins with Sialic Acid-Specificity

The purpose of this work is to perform a detailed study of carbohydrate specificity of the new extracellular bacilli lectins which is considered to determine mechanisms of the lectins action. Sources of lectins were bacterial strains from Ukrainian collection of microorganisms. The optimized protocol of bacilli lectins isolation and purification included precipitation with ammonium sulfate with subsequent gel filtration chromatography on Sepharose CL-6B. Hemagglutinating activity of bacilli lectins and their fine carbohydrate specificity to sialic acids and their derivatives as well as sialic asid-containing and asialic glycoconjugates were studied. The ability of extracellular bacilli lectins to discriminate a- and 13-conformation of carbohydrate molecule and the type of connection between the monomers was determined. Studied lectins showed the most affinity to glycoconjugates containing both types of sialic acids （N-acetylneuraminic acid （Neu5Ac） and N-glycolylneuraminic acid （NeuGc）） and it is supposed to be a basis of their diagnostic and analytical potential.     

Toxicity of Bacillus thuringiensis Crystal Toxins to Field Populations of Rice Leaf Folder, Cnaphalocrocis medinalis （Guenee） and Establishment of Baseline Susceptibility to Cry1Ab

Eight insecticidal crystal proteins of Bacillus thuringiensis, CrylAa, CrylAb, CrylAc, CrylB, Cry2Aa, CrylC, CrylDa and Cry 1Ea were assessed for toxicity against 1 st instar larvae of rice leaf folder, Cnaphalocrocis medinalis （Guenee） at 48 HAT and 72 HAT. Bioassay results depicted CrylAa was the most toxic （LCso 2.35 ppm） followed by CrylBa （LCso 8,50 ppm） and CrylAb （LCso 8.73 ppm） at 48 HAT, whereas, at 72 HAT CrylAb proved to be highly toxic （LC50 0.50 ppm） followed by CrylAa （LCso 4.07 ppm）, CrylAc （LCso 4,84 ppm） and CrylBa （LCso 6.42 ppm）. Toxins Cry2Aa, CrylCa, CrylDa and CrylEa did not resulted in any mortality at 48 HAT and 72 HAT, respectively. Baseline estimates for CrylAb against 1st instar larvae of C. medinalis sampled from seven geographical locations revealed variation in LC50＇s from 0.37 ppm to LC50 16.25 ppm at 48 HAT and LC50 0.50 ppm to LC50 6.49 ppm 72 HAT, respectively with relative resistance ratios of 44-fold and 13-fold at 48 HAT and 72 HAT over the susceptible population.     

Risk Assessment and Biosafety Studies of Transgenic Bt Rice （Oryza sativa L.）

Transgenic crops having alien genes from different sources are getting popularity. A Rice （Oryza sativa L.） containing gene from Bacillus thuringiensis, a soil bacterium, was assessed to study the potential risks of transgenic plants on environment. The crop was found resistant to target insect pests. Rice variety Basmati-370 transformed with two insecticidal genes, crylAc and cry2A, was grown under field conditions for several years. Data were collected at different stages of the plant growth. Fate of Cry protein in soil, effect of Bt protein on non-target insects, risks of vertical and horizontal gene flow were evaluated. No potential hazard was found at all levels. Bt protein was unstable and degraded significantly in soil within 30 days after harvesting the crop. No harmful effects were found on non-target insects （insects other than order lepidoptera）. Maximum gene flow of 0.02% was observed at close spacing and no evidence of horizontal gene transfer to Rhizobium spp. was found. In conclusion, the transgenic rice plants transformed with Bt genes have no harmful effects on the environment.     

Bacillus subtilis fmbR抗菌物质的分离和鉴定

 【目的】对Bacillus subtilis fmbR产生的抗菌物质进行分离和鉴定研究，以确定抗菌物质的组成和结构。【方法】采用HPLC和TLC层析对Bacillus subtilis fmbR抗菌物质进行分离纯化，通过ESI-MS和MALDI-MS分析对抗菌物质的组成和结构进行初步鉴定。【结果】HPLC层析表明了Bacillus subtilis fmbR抗菌物质含有保留时间与surfactin相似的成分。TLC层析和原位酸解证明了Bacillus subtilis fmbR抗菌物质含有闭合肽键类的物质，其中之一为相对迁移率Rf与标样surfactin相近的组分。采用ESI-MS分析检测到Bacillus subtilis fmbR抗菌物质含有分子量与surfactinA相同的m/z1009.1、m/z1023.2 和m/z1037.0等3种同系物；通过MALDI-MS分析获得[M+H]+为m/z 3403.95抗菌物质，该物质分子量与Bacillus subtilis 168产生的细菌素subtilosin的m/z3403.3 相同。【结论】Bacillus subtilis fmbR抗菌物质由C13～C15的3种surfactinA同系物和一种羊毛硫抗生素subtilosin组成。     

Study on regulation of potassium loss and its balance characteristics from typical paddy soils in Dongting Lake area

Potassium loss due to runoff and leaching, crop uptake, and K-fertilizer input, irrigation and wet deposition was studied, subsequently potassium balance characteristics was analyzed using lysimeter modeled two typical paddy soils from double rice field in Dongting Lake area. K losses due to runoff and leaching in Alluvial Sandy Loamy Paddy Soil （ASL） with lower cation exchange content （CEC） and clayey granule content were increased significantly compared with those in Reddish Yellow Loamy Paddy Soil （RYL） with higher CEC and clayey granule content. Compared with RYL, lower economic and biological yield due to lower capacity of nutrient and water keeping in ASL resulted in lower K uptake. So potassium deficit in double rice field of RYL was more serious than that of ASL because main K proportion was accounted by crop uptake. Available K in soil of surface layer （0-15 cm） was decreased significantly due to long term nutrient management of K deficit in double rice field. The straw return paddy field was suggested as an effective way to increase soil K and balance K deficit in double rice field because large amount of potassium was enriched in rice straw at harvest.     

Transgenic cry1Ab/vip3H＋epsps Rice with Insect and Herbicide Resistance Acted No Adverse Impacts on the Population Growth of a Non-Target Herbivore,the White-Backed Planthopper,Under Laboratory and Field Conditions

Numerous Bt rice lines expressing Cry protein derived from Bacillus thuringiensis Berliner （Bt） have been developed since 1989. However, the potential risks posed by Bt rice on non-target organisms still remain debate. The white-backed planthopper （WBPH）, Sogatella furcifera （Horváth）, is one of the most economically important insect pests of rice in Asian countries and also one of the main non-target herbivores of transgenic rice. In the current study, impacts of transgenic cry1Ab/vip3H＋epsps rice （G6H1） with both insect and herbicide resistance on WBPH were evaluated to ascertain whether this transgenic rice line had potential risks for this sap-sucking pest under laboratory and ifeld conditions. The laboratory results showed that no signiifcant difference in egg developmental duration, nymphal survival rate and female fecundity was found for WBPH between G6H1 and its non-transgenic isoline （XS110）. However, the development duration of nymphs was signiifcantly shorter and female longevity signiifcantly longer when WBPH fed on G6H1 by comparison with those on its control. To verify the results found in laboratory, a 3-yr ifeld trial was conducted to monitor WBPH population using both the vacuum-suction machine and beat plate methods. Although the seasonal density of WBPH nymphs and total density of nymphs and adults were not signiifcantly affected by transgenic rice regardless of the sampling methods, the seasonal density of WBPH adults in transgenic rice plots was slightly lower than that in the control when using the vacuum-suction machine. Based on these results both from laboratory and ifeld, it is clear that our tested transgenic rice line will not lead higher population of WBPH. However, long-term ifeld experiments to monitor the population dynamics of WPBH at large scale need to be conducted to conifrm the present conclusions in future.     

Effects of Elemental Sulphur and Farmyard Manure on pH and Salinity of Calcareous Sandy Loam Soil and Some Nutrient Elements in Tomato Plant

The objective of this study was to evaluate the effects of elemental sulphur （S） and farmyard manure on soil pH, EC and N, S, P concentrations of tomato grown in a calcareous sandy loam soil. For this purpose, a pot experiment was conducted in greenhouse conditions. Sulphur was applied at 0, 50, 100, 150, 200, 400 mg kg~ and farmyard manure at 0, 3 ton da^-1 to the soil. Three weeks after applications, tomato seedlings were planted and 8 weeks later, the plants were harvested to determine N, S, P concentrations and dry matter yield. Soil pH and EC were determined in the soil samples taken at 3 different periods. Effects of sulphur and farmyard manure applications were not significant on N, P concentrations and dry matter yield of tomato plant. S concentration of tomato plant was increased by sulphur alone. Soil pH was decreased and soil EC was increased in both 2^nd and 3^rd soil sampling period by the sulphur applications. As a result of farmyard manure application, soil pH decreased in the 2^nd soil sampling period but increased in the 3^rd soil sampling period. Also, soil EC was significantly increased in 1^st and 3^rd soil sampling period. Farmyard manure had no significant effect on S supply to tomato plant. The N：S ratio of tomato plant was decreased by sulphur alone. However, the sulphur with farmyard manure applications decreased N：S ratio in lesser extend compared to the S applications.     

Application of Neutron Activation Analysis Technique for the Analysis of Soil Samples from Farmlands of Yebrage Hawariat,East Gojjam,Ethiopia

Farmers may not be conscious for their farmland＇s nutrients, soil organic matter, water and air because they simply concerned only for their labor availability and soil fertility losses. The composition and proportion of these components greatly influence soil physical properties, including texture, structure and porosity, the fraction of pore space in a soil. The soil of this farmland must be able to supply adequate amount of plant nutrients, in forms which can be absorbed by the crop, within its lifespan. Deficiencies or imbalances in the supply of any of essential elements can compromise growth, affecting root development, cell division, crop quality, crop yield and resistance to disease and drought. This study was conducted to fill this knowledge gap in order to develop economically vital and environmentally accepted nutrient management strategies for the use of soils in agricultural lands. The objective of this study is to assess the elemental contents and concentration of soil samples collected from farmlands of ＂Yebrage＂ using neutron activation analysis （NAA） techniques regardless of oxidation state, chemical form or physical locations. NAA is used to determine the elemental composition and concentrations present in a soil. The macro/micronutrient and organic matter deficiencies have been verified in agricultural soils through increased use of soil testing and plant analysis. The challenge for agriculture over the coming decades will meet the world＇s increasing demands for food in a sustainable way. Current issues and future challenges point out that as long as agriculture remains a soil based industry, major decreases in productivity likely to be attained ensuring that plants do not have adequate and balanced supply of nutrients.     

Effects of Various Degrees of Antennal Ablation on Mating and Oviposition Preferences of the Diamondback Moth,Plutella xylostella L.

Using scanning electron microscopy, we investigated the distribution of the trichoid, basiconic, and coeloconic sensilla on the antennae of the diamondback moth （DBM;Plutella xylostella）. The trichoid sensilla were the most abundant sensory organ, and the male moth antennae host signiifcantly more trichoid sensilla than female moth antennae. Conversely, basiconic and coeloconic sensilla were found more frequently on female than on male antennae. We performed experiments with various degrees of antennal ablation and demonstrated that DBM antennae played a key role in the control of mating and oviposition. We found that neither oviposition preference nor mating behaviors changed signiifcantly when less than 1/4 of both antennae were removed. However, there was a signiifcant behavioral change when the antennae were ablated by more than half. As the length of the antenna was shortened, the successful mating rate decreased and mating peak was delayed. An otherwise consistent host preference for oviposition was eliminated when both antennae were completely removed. Furthermore, we found that the number of trichoid sensilla was positively correlated with mating rate and oviposition preference. However, the numbers of basiconic and coeloconic sensilla were not correlated with mating rate and mating peak, but highly correlated with oviposition preference. Taken together, our results indicate that antennal sensory information plays a critical role in the mating and oviposition behaviors of this economically important pest.     

Hydrolytic Enzyme Inhibitors and Necrotic Reactions in Potato Leaves Reduce Reproductive Success of Colorado Potato Beetle

In field and laboratory conditions the reproductive potential （survival at different stages of development） of the Colorado potato beetle has been measured, the dependence of the parameters of insects fertility on varietal features of the feed has been investigated, the food activity of the representatives of insects from four areas of the Southern Urals has been measured, the intensity and time of hypersensitivity reactions on potato leaves has been defined. In laboratory experiments, the level of activity of hydrolytic enzymes in insects and the level of activity of these enzymes inhibitors in the leaves of three potato varieties differing in their resistance to the Colorado potato beetle have been determined. The results show that the insects-representatives of different samples have different food activity in relation to different potato varieties. High mortality rates were revealed at feeding with leaves of potato of the Bashkir variety characterized by a high level of hydrolases inhibitors and relatively rapid development of necrotic reactions after the appearance of eggs laying on the leaves. We suppose that the neerogenetic barrier appearing in the form of necrosis is the major cause of the fetal mortality of insects. The level of the content of inhibitors inactivating larvae digestive enzymes affects the survival of insects at postembryonic stages. The speed of the necrosis emergence, the level of activity of digestive enzymes inhibitors in potato can be considered as criteria for the selection of breeding material for resistance to the Colorado potato beetle.     

Distribution and Identification of Ixodid Tick Species on Livestock in Northern Pakistan

Identification of lxodid tick species on livestock in two districts, viz., D.I. Khan and Lakki Marwat of Northern Pakistan were made during the period from March to November 2009 through animals＇ survey and questionnaire. The main tick species were Amblyomma （14.3%）, Boophilus microplus （l 8.5%） and Hyalomma anatolicum and H. dromedarii （67.2%）. Out of 3009 collected ticks, the highest number of female （88.7%） and the lowest of males （ l 1.3%） belong to Boophilus; the lowest number of female （92.4%） and the highest of males （77.5%） belong to Amblyomma. However, cows were more susceptible than buffaloes and camels ranked third to ticks infestation. Moreover, cows, buffalos, goats and sheep harbored mixed infestation （had more than one type of ticks）, however, camels and donkey harbored single infestation （had only one type of tick）. An emerging ethno-veterinary or traditional remedy is poultry keeping for ticks＇ elimination in Lakki Marwat. The results of this research will be helped in taking appropriate measures to reduce tick infestation and to improve management practices. Hence, it will help to increase farm productivity and farmers income.     

Probabilistic Model of the Brown Rat Control

We represent assessment of the rats control operator＇s actions, starting from the placement of rat control means （chemical, mechanical and others） in the object territory until the full its elimination and followed by assessment of the probability of rat population recovery. The probability of success is evaluated when using a combination of rat control means. We took into account changes in rat population occurring in different calendar periods of the year. The proposed calculation method can be used in training programs, as well as for the local forecast of releasing objects from rats and rats＇ re-settling.     

Tillage and Poultry Manure Effects on Soil Physical Properties,Nutrient Status,Growth, Dry Matter and Grain Yield of Sorghum

The effects of different tillage systems and poultry manure on soil physical properties, performance and nutrients in sorghum were studied for three years at Owo, southwest Nigeria. There was factorial combinations of herbicide-based zero tillage （ZT）, manual clearing （MC）, disc ploughing （P）, ploughing plus harrowing （P＋H） and ploughing plus double harrowing （P＋2H）, and two rates of poultry manure at 0 and 7.5 Mg ha^-1. Herbicide-based zero tillage and manual clearing reduced soil temperature and conserved more water than mechanized tillage techniques. Poultry manure reduced soil bulk density and temperature and increased soil water and porosity. There was a percentage decrease of leaf N, P, K, Ca and Mg concentrations, plant height, leaf area, stem girth, root dry weight, dry matter and grain yield in ascending order for herbicide-based zero tillage, manual clearing, ploughing, ploughing plus harrowing and ploughing plus double harrowing while percentage increases were recorded in a descending order for all the various combinations of tillage with poultry manure in that order. Poultry manure in combination with tillage increased dry matter and grain yield by 33.1 and 39.5%, respectively in comparison with tillage only. The manure-zero tillage methods increased dry matter and grain yield by 8% and 15%, respectively when compared with manure-mechanized tillage methods. Zero tillage or manual clearing in combination with 7.5 Mg ha^-1 poultry manure was most suitable for sorghum cultivation.