Responses of Anise Medicinal Plant Species in Terms of Essential Oil Contents and Concentrations to Different Planting Times and Various Nitrogen Fertilizer Sources under Semi-Arid Climatic Conditions

Essential oil content and concentration of anise plant planted in different times under various sources of nitrogen was evaluated in this study. The experiment was conducted in a split plot based on a randomized complete block (RCB) design (different sources of fertilizer as the main plot and planting dates as subplots) with three replications in the Agricultural Research Station at Ferdowsi University of Mashhad in 2015–2016. Treatments included: planting times (sub plot factor): (27^th of October, 27^th of November, 1^st of March, 10^th of March, and 8^th of April) and nutritional sources (main plot factor): (Cow Manure, Chemical Fertilizer, and mixture of Cow Manure + Chemical Fertilizer). The results showed that the anise plant essential oil qualitative characteristics were significantly influenced by treatments. Also, interaction between the fertilizer treatments and planting dates on the essential oil content and concentration of the plants were significant. The highest percentage of plant essential oil (3.71%) was found in the cow manure treatment, and the lowest percentage of essential oil (2.95%) was obtained in the chemical fertilizer. The highest essential oil yield (16.09 kg/ha) was found in the mixed treatment of the cow manure and the chemical fertilizer, and the chemical fertilizer treatment had the lowest rate of essential oil yield (11.41 kg/ha). The highest percentages of the plant oil (14.31%) and the plant oil yield (77.46 kg/ha) were found in the integrated treatment of the cow manure and the chemical fertilizers. The lowest amounts of the plant oil (12.19%) and the plant oil yield (41.28 kg/ha) were found in the chemical fertilizer. According to the percentage and the yield of the essential oil, the best planting time and fertilizer treatment was the one on 10th of March and using integrated cow manure and chemical fertilizer.     

DNA vaccine encoding avian influenza virus H5 and Esat-6 of Mycobacterium tuberculosis improved antibody responses against AIV in chickens

The H5 gene of avian influenza virus (AIV) strain A/chicken/Malaysia/5744/2004(H5N1) was cloned into pcDNA3.1 vector, and Esat-6 gene of Mycobacterium tuberculosis was fused into downstream of the H5 gene as a genetic adjuvant for DNA vaccine candidates. The antibody level against AIV was measured using enzyme-linked immunosorbent assay (ELISA) and haemagglutination inhibition (HI) test. Sera obtained from specific-pathogen-free chickens immunized with pcDNA3.1/H5 and pcDNA3.1/H5/Esat-6 demonstrated antibody responses as early as 2 weeks after the first immunization. Furthermore, the overall HI antibody titer in chickens immunized with pcDNA3.1/H5/Esat-6 was higher compared to the chickens immunized with pcDNA3.1/H5 (p < 0.05). The results suggested that Esat-6 gene of M. tuberculosis is a potential genetic adjuvant for the development of effective H5 DNA vaccine in chickens.     

Comparison of the pathogensis of two isolates of <Emphasis Type="Italic">Besnoitia caprae</Emphasis> in inbred BALB/c mice

This study was performed to evaluate the infectivity of bradyzoites of two Besnoitia caprae isolates, BC-1 and BC-2, to inbred BALB/c mice. Each group of inbred BALB/c mice was inoculated intraperitoneally with 1 × 10³, 1 × 10⁴, 1 × 10⁵, 5 × 10⁵ and 1 × 10⁶ of one of the two isolates of B. caprae bradyzoites. The mice were monitored daily for a period of 40 days for survival. After death of each mice, several passages from its peritoneal washing and tissues were analyzed using ribosomal DNA-specific PCR assay. Marked differences in pathogenicity between the isolates were seen. All the inbred BALB/c mice infected with BC-2 survived but all the mice that were administered with 1 × 0⁵, 5 × 10⁵ and 1 × 10⁶ BC-1 bradyzoites were died within 4–9 days post-infection (DPI). Histopathological examination of the tissues of the dead mice revealed hyperemia and necrosis with presence of mononuclear and polymorphonuclear cell infiltration in myocardium, spleen and intestines together with interstitial pneumonia and peritonitis. All inbred BALB/c mice in the 1 × 10³ and 1 × 10⁴ groups of BC-1 inoculated mice survived and they were euthanized after 40 DPI. Chronic inflammation with infiltration of mononuclear cells was evident in myocardium, spleen, alveolar septa of the lungs of most of the examined tissues with hemorrhagic enteritis in the mice infected with 1x10⁶ bradyzoites. The mice infected with different doses of BC-2 were euthanized after 40 DPI and no lesion was seen in histopathological sections of their organs. All peritoneal washings and examined tissues were PCR positive in BC-1 group. This experiment is the first report to show inbred BALB/c mice as a relevant model for B. caprae and demonstrates that this strain of inbred BALB/c mice is a suitable animal model for biological studies and examination of pathogenesis for this species of Besnoitia. The present findings also provide evidence for significant differences between the two isolates of B. caprae.     

Induction of Immunogenic Response in BALB/c Mice by Live and Killed Form of Recombinant Lactococcus lactis Displaying EG95 of Echinococcus granulosus

Background:CE is a zoonotic parasitic infection caused by Echinococcus granulosus worldwide and is associated with economic losses among livestock animals. EG95 is an immunogenic antigen from the E. granulosus. Lactococcus lactis has been prested as a safe vehicle for antigen delivery. The goal of this study was to design a novel L. lactis strain displaying EG95 as a vaccine delivery system. Methods:The eg95 encoding gene fragment fused to the M6 anchoring protein was cloned into the pNZ7021 vector, and L. lactis NZ9000 displaying recombinant EG95 was constructed. The expression of an approximately 32-kDa EG95 protein was confirmed by Western blotting and immunofluorescence analysis. The immune responses were evaluated in BALB/c mice immunized orally and subcutaneously with the live and killed recombinant L. lactis, respectively. Results:Total IgG level in mice immunized with heat-killed recombinant L. lactis (pNZ7021-eg95) significantly increased compared to the control group. sIgA was significantly higher in mice received live recombinant L. lactis (pNZ7021-eg95) compared to the control mice. Splenic lymphocytes from immunized mice represented the high levels of IFN-γ and the low-levels of IL-4 and IL-10. Conclusion:Our results indicate that immunization with EG95-expressing L. lactis can induce both specific humoral and cellular immune responses in mice. Key Words: Echinococcus granulosus, Lactococcus lactis, Immunization, Vaccines     

Foliar Nitrogen Uptake from Wet Deposition and the Relation with Leaf Wettability and Water Storage Capacity

This study assessed the foliar uptake of ¹⁵N-labelled nitrogen (N) originating from wet deposition along with leaf surface conditions, measured by wettability and water storage capacity. Foliar ¹⁵N uptake was measured on saplings of silver birch, European beech, pedunculate oak and Scots pine and the effect of nitrogen form (NH ₄ ⁺ or NO ₃ ^? ), NH ₄ ⁺ to NO ₃ ^? ratio and leaf phenology on this N uptake was assessed. Next to this, leaf wettability and water storage capacity were determined for each tree species and phenological stage, and the relationship with ¹⁵NH ₄ ⁺ and ¹⁵NO ₃ ^? uptake was examined. Uptake rates were on average five times higher (p?<?0.05) for NH ₄ ⁺ than for NO ₃ ^? and four times higher for deciduous species than for Scots pine. Developing leaves showed lower uptake than fully developed and senescent leaves, but this effect was tree species dependent. The applied NH ₄ ⁺ to NO ₃ ^? ratio did only affect the amount of N uptake by senescent leaves. The negative correlation between measured leaf contact angles and foliar N uptake demonstrates that the observed effects of tree species and phenological stage are related to differences in leaf wettability and not to water storage capacity.     

Differentiation of human umbilical cord mesenchymal stem cell into germ-like cell under effect of co-culture with testicular cell tissue

Supplements produced by mouse testicular cells (mTCs) and the interaction between cells can increase the differentiation rate of human umbilical cord mesenchymal stem cells (hUCMSCs) into the germ-like cells. We studied the differentiation rate of hUCMSCs into the germ-like cells under effect of mTCs co-culturing. Isolated hUCMSCs from postpartum human umbilical cords were cultured. Then, the expression of mesenchymal (CD73, CD90 and CD105) and haematopoietic (CD34 and CD45) markers of hUCMSCs were confirmed by flow cytometry. Then, the hUCMSCs were cultured in four distinct groups: (a) control, (b) co-culture until D0, (c) co-culture until D5 and (d) co-culture until D10, in order to differentiate into the germ-like cells. After 10 days, the expression of OCT4, VASA, Fragilis and SYCP3 genes were examined by Real-Time qPCR. The flow cytometry indicated a high expression of mesenchymal markers and a low expression of haematopoietic markers (CD73:98.6%, CD90: 99.1%, CD105: 99.5%, CD34: 4.22% and CD45: 2.54%). The expression of OCT4 decreased during the time while the expression of VASA, Fragilis and SYCP3 markers increased in the co-culture with testicular cells (p value <.05). Co-culture with mTCs may be used as an effective method to differentiate hUCMSCs into germ-like cells.     

Digital assessments of soil organic carbon storage using digital maps provided by static and dynamic environmental covariates

Understanding the vertical and lateral distribution of soil organic carbon (SOC) and soil organic carbon density (SOCD) is indispensable for soil use and environmental management because of their vital role in soil quality assessments. Primarily, they are needed in calculating soil organic carbon storage (SOCS). The objective of this research was to provide digital maps of SOC and SOCD variation as well as their uncertainties at multiple standardized depths (H1: 0–5, H2: 5–15, H3: 15–30, H4: 30–60 and H5: 60–100 cm) using a parsimonious model with optimized terrain-related attributes and satellite-derived data. SOCS were evaluated at soil subgroup levels. An area of about 808 km² with varying elevation, plant cover and lithology from the Miandoab region, West Azerbaijan Province, Iran was selected as a case study area. A total of 386 soil samples were collected from 104 profiles comprising various soil genetic horizons. A continuous spline function was then fitted to the target properties in advance of creating a dataset at five standard depth intervals (following the GlobalSoilMap project). These were then grouped into three classes including top (H1), middle (H2, H3 and H4) and bottom (H5) depths to ease interpretation. Static and dynamic covariates (30-m resolution) were derived from a digital elevation model (DEM) and a suite of Landsat-8 spectral imageries, respectively. Four candidate models including stepwise multiple linear regression (SMLR), random forest (RF), cubist (CU) and extreme gradient boosting (XGBoost) Tree were tested in this study. Finally, the digital maps at 30-m resolution of SOC and SOCD and their uncertainties were prepared using the best-fit model and the bootstrapping method, respectively. Four soil subgroups (Gypsic Haploxerepts, Typic Calcixerepts, Typic Haploxerepts and Xeric Haplocalcids) were identified across the study area. The covariates had variable contributions on the evaluated models. The XGBoost Tree model generally outperformed other models for prediction of SOC and SOCD (R² = 0.60, on average). Regardless of soil subgroups, the uncertainty analysis showed that the SOCD map had a low prediction interval range value indicating high accuracy. Additionally, the highest SOCS and SOCD was observed at the top followed by middle and bottom depths in the study area. All subgroups exhibited a decreasing trend of SOCD with increasing depth. A similar trend was also observed for SOCS. The highest SOCD (on average) was observed in Gypsic Haploxerepts (4.71 kg C/m²) followed by Typic Calcixerepts (4.46 kg C/m²), Typic Haploxerepts (4.45 kg C/m²) and Xeric Haplocalcids (4.40 kg C/m²). Overall, the SOCS normalized by area within soil order boundaries was greater in Inceptisols than Aridisols across the study area. The findings of this study provide critical information for sustainable management of soil resources in the area for agricultural production and environmental health in the Miandoab region of Iran.     

Comparison of artificial neural networks and prediction models for reference evapotranspiration estimation in a semi-arid region

Estimation of reference evapotranspiration (ET_o) is essential for determination of crop water requirements. In this research, Penman–FAO (P-FAO) and Penman–Monteith (PM) equations were calibrated and validated by lysimeter-measured ET_o with six and four weather parameters. Furthermore, two input structures (six and four weather parameters) to artificial neural networks (ANNs) were investigated. Results showed that the accuracy of the PM equation is greater than that of the P-FAO equation. An empirical equation was developed to estimate daily ET_o using mean daily temperature and relative humidity, and sunshine hours. The accuracy of the equation to estimate daily ET_o using smooth weather data is greater than that of an equation using original data. Furthermore, ANNs were able to estimate ET_o properly. The accuracy of ANNs with six inputs is higher than that obtained using the P-FAO equation and is similar to that determined using the PM equation. A decrease in number of inputs to ANNs generally decreased the accuracy of estimation, however, ANNs were able to estimate ET_o properly when wind speed and solar radiation were unavailable. Furthermore, the accuracy of ANNs, with four input parameters is greater than that obtained using the PM equation and is similar to that obtained with P–FAO and the developed empirical equations.     

Compost Amendment to Sandy Soil Affects Soil Properties and Greenhouse Tomato Productivity

Sandy soils, with low productivity, could be improved by compost application to sustain crop production. This study aimed to examine the effect of three compost types (vegetable, fruit and yard waste compost, garden waste compost, and spent mushroom compost) on basic properties of a loamy sand and greenhouse tomato productivity. Disturbed and intact soil samples were taken from a decade-long compost field experiment on loamy sand with three compost types at application rate of 30 m³ ha^?1 yr^?1 (7.5 ton ha^?1 yr^?1). The soils were characterized for chemical and physical properties. Tomato was planted in a greenhouse using soil samples from the field and vegetative and yield parameters (plant height, stem diameter, leaf number, and fruit yield), water productivity, and harvest index were evaluated. All compost types significantly increased soil total carbon, total nitrogen, pH, electrical conductivity and significantly decreased bulk density, with no effect on plant available water compared to the control. Fresh and dry fruit weights were significantly increased after compost addition. Plant height, leaf number, stem diameter, and total biomass did not significantly improve after compost addition. Spent mushroom compost had greater effect in improving tomato productivity. A decade-long application of composts on loamy sand improved basic chemical and physical properties which were reflected in increased fruit yield in tomato. Since no negative effect of compost was observed, we suggest that sandy soils may serve as a safe end use option for these composts and potentially support crop growth.     

Soil application of entomopathogenic nematodes suppresses the root-knot nematode Meloidogyne javanica in cucumber

Journal of Plant Diseases and Protection - Cucumber (Cucumis sativus) cultivation in commercial greenhouses occupies an important section of vegetable production in Iran. Root-knot nematode...