Cellular Immune Responses to 35 kDa Recombinant Antigen of Mycobacterium avium paratuberculosis

Mycobacterium avium paratuberculosis is the causative agent of Johne disease, a chronic ulcerative intestinal condition in ruminant animals. Owing to the predominance of cellular response in subclinical forms of the infection, identification of M. a. paratuberculosis antigens eliciting host cell-mediated immune (CMI) reaction is crucial for early control of the disease. A 35 kDa protein of M. a. paratuberculosis was studied for its ability to elicit CMI responses using a mouse model. Lymphoproliferation and IFN-γ response were used to measure the CMI response. Recombinant 35 kDa protein (P35) stimulated proliferation of mouse mononuclear splenocytes sensitized with M. a. paratuberculosis. The P35 elicited increased nitrite production from mononuclear splenocytes from M. a. paratuberculosis-sensitized mice. In addition, RT-PCR-based semiquantitative IFN-γ measurement showed that stimulation with P35 is associated with significant expression of IFN-γ mRNA in M. a. paratuberculosis-sensitized mouse splenocytes. The results indicate that the 35 kDa protein of M. a. paratuberculosis is associated with CMI response in the host.     

Preliminary investigations of the distribution of Escherichia coli O149 in sows, piglets, and their environment

Little is known about the sources and kinetics of enterotoxigenic Escherichia coli colonization in pigs during the pre-and post-weaning period. In this study, farrowing pens, sows, and piglets were tested for the presence of E. coli O149 by real-time polymerase chain reaction (PCR) after bacterial culture pre-enrichment on 2 farms, one with a history of post-weaning diarrhea (problem farm - PF) and the other without such a history (non-problem farm - NPF). Unlike those on the PF, the sows from the NPF did not carry E. coli O149 before parturition, although they were colonized to frequencies similar to animals on the PF soon afterwards. Most piglets from the NPF were colonized within a week after birth, whereas only a small proportion of those on the PF were colonized during that period. No difference was observed in the frequency of piglet colonization at the 2 farms either at weaning or during the following week. Post-weaning diarrhea (PWD), which is caused by enterotoxigenic E. coli (ETEC), is a multifactorial disease. The presence of ETEC alone is not always sufficient for the disease to develop. Many other factors are considered to be associated with the occurrence of PWD, including feed type (1,2), feeding regimen (1,3,4), the presence of other infectious agents (3,5), weaning age, and weight (6). Weaning, which is considered to be a major physiological and psychological stress factor, is critical for the disease to occur (7). Although piglets are already colonized with ETEC before weaning (4,8), on many farms, clinical disease occurs only after weaning (1). Both sows (9,10) and the environment (6) could be possible sources of infection for piglets, but results from previous studies have not resolved this issue because of the low sensitivity of ETEC detection methods. This study provides preliminary data based on a sensitive detection method for E. coli O149 in pigs and their environment. The results demonstrate the potential of real-time PCR for future studies on this topic.     

Entomotoxic effect of silica nanoparticles against <Emphasis Type="Italic">Sitophilus oryzae</Emphasis> (L.)

Fumigants and residual insecticides are commonly used to combat stored grain pests. In recent years, consumer awareness of the health hazard from residual toxicity and the growing problem of insect resistance to these conventional insecticides have led the researchers to look for alternative strategies for stored grains protection. For example, diatomaceous earth (DE) can be effective against stored grain insects. In this study, DE was used to design amorphous nano sized hydrophilic, hydrophobic, and lipophilic silica in 15–30 nm size range. Nanocides are expected to reduce the volume of application and kinetics of development of resistance in pests. We hypothesized that surface-functionalized silica nanoparticle (SNP) might be a viable alternative to conventional pesticides. Entomotoxicity of SNP was tested against rice weevil Sitophilus oryzae and its efficacy was compared with bulk-sized silica (individual particles larger than 1 μm). Amorphous SNP was found to be highly effective against this insect pest causing more than 90% mortality, indicating the effectiveness of SNP to control insect pests.     

Short-lived nuclides in hibonite grains from Murchison: evidence for solar system evolution

Records of now-extinct short-lived nuclides in meteorites provide information about the formation and evolution of the solar system. We have found excess 10B that we attribute to the decay of short-lived 10Be (half-life 1.5 million years) in hibonite grains from the Murchison meteorite. The grains show no evidence of decay of two other short-lived nuclides-26Al (half-life 700,000 years) and 41Ca (half-life 100,000 years)-that may be present in early solar system solids. One plausible source of the observed 10Be is energetic particle irradiation of material in the solar nebula. An effective irradiation dose of approximately 2 x 10(18) protons per square centimeter with a kinetic energy of >/=10 megaelectronvolts per atomic mass unit can explain our measurements. The presence of 10Be, coupled with the absence of 41Ca and 26Al, may rule out energetic particle irradiation as the primary source of 41Ca and 26Al present in some early solar system solids and strengthens the case of a stellar source for 41Ca and 26Al.     

Stammapplikationen im Obstbau mit nanostrukturierten Silikaten gegen die Blutlaus Eriosoma lanigerum (Hausm.) und den Gemeinen Ohrwurm Forficula auricularia L.

The present work deals with a new strategy to control Eriosoma lanigerum (Hausm.) in apple and Forficula auricularia?(L.) in apricots. With common insecticides, both pests are difficult to control. Although F.?auricularia is regarded as a beneficial insect, in ecological agriculture it has the potential to cause severe economical damage in apricots. In apple and apricot orchards we tested treatments on the bark with diatomaceous earth (DE) products to prevent insects from reaching the tree crown. DE has been applied as liquid (“Fossil Shield^®” translucend) by a backpack sprayer or electrostatically as powder (“Fossil Shield^®” 90.0?s). In some trials we covered the treated area with a plastic funnel (fruitcap) in order to protect the silica-product from wind and rain. Experiments were evaluated weekly and number of insects in the tree crown was counted. During the first two months after treatment E.?lanigerum population in the crown was reduced by 98.2%. However, over the whole period of six month the barriers turned out to be ineffective (p?=?0.617). Winged adults have been capable to pass the barriere. In contrast to this fruitcap barriers were able to keep F. auricularia away from the crown (p?=?0.000). They have been as effective as standard treatment using glue. In such treatments apricot yield increased 6-fold for the cultivar “Goldrich” and 5.5-fold for the cultivar “Hargrand”.     

Modeling of abrasion resistance of Persian handmade wool carpets using response surface methodology

The effect of knot density, pile height, number of ply in pile yarn and pile yarn twist on abrasion resistance of Persian handmade wool carpets has been studied. The interaction between the process variables has been analyzed by using response surface methodology based on the Box-Behnken design of experiment. Knot density, pile height, interactions between knot density and pile height; pile height and pile yarn twist; as well as squares of the knot density, pile height and number of ply in pile yarn are significant process variables. The minimum abrasion loss occurs at the combination(s) of medium values of knot density (six knots per inch) and number of ply in pile yarn (three ply) as well as lower values of pile height (ten mm) and pile yarn twist (three and a half twists per inch).     

Immunopathological effects of experimental T-2 mycotoxocosis in broiler chicken co-infected with infectious bronchitis virus (IBV)

A total of 128 1-week-old chicks were classified into four groups; T-2 toxin fed (T-2), IBV infected (IBV), T-2 toxin fed and co-infected with IBV (T-2+IBV), and untreated (control) for a period of 6 weeks. Within their respective groups, the birds belonged to T-2 and T-2+IBV were exposed to 2 ppm of T-2 toxin contaminated feed for 6 weeks, and 0.2 ml of 10 EID(50) (10(5.69)/0.2 ml) inoculums of IBV isolate (India/LKW/56/IVRI/08) was used to challenge the chicks belonged to IBV alone and T-2+IBV groups after 3 weeks of the experiment. To study immunopathological effects, parameters such as lymphocyte stimulation indices (SI), haemagglutination inhibition, enzyme linked immunosorbant assay (ELISA), peripheral lymphocytes CD(4)(+) and CD(8)(+) analysis, and histopathological examination of lymphoid organs were done. Accordingly, SI values were significantly (P<0.05) lower in all the treatment groups as compared to control, however, the SI values of IBV infected group were significantly higher than the values in toxin fed groups. The mean HI titres to ND vaccine was significantly (P<0.05) lower in the toxin groups at all the intervals, and the antibody titres in IBV infected group were significantly (P<0.05) higher than that of T-2 toxin fed and co-infected with IBV group but were significantly (P<0.05) lower than the control at 21 (3) and 28 (10) days of toxin feeding (DTF) (days post infection (DPI)). Similarly, the mean IBV ELISA antibody titres in the toxin fed groups were significantly (P<0.01) reduced as compared with the IBV ELISA antibody titres of IBV infected but not toxin fed group, at all intervals. Peripheral CD(4)(+):CD(8)(+) ratios in T-2+IBV group and number of CD(4)(+) and CD(8)(+) peripheral lymphocytes in all treatment groups were significantly reduced as compared to the values in control birds. However, CD(4)(+):CD(8)(+) ratios of IBV infected group at 42 (21) DTF (DPI) were found significantly (P<0.05) higher than the values in control birds. Histopathologically, lymphoid organs (bursa of Fabricius, spleen, thymus, caecal tonsils and Harderian glands) showed moderate to severe necrosis (lymphocytolysis) and extensive lymphocyte depletion in all the toxin groups (T-2 and T-2+IBV groups) where the severity and extent of the lesions were more in T-2+IBV group. The findings of the present experiment revealed immunosuppressive effects of T-2 toxin and aggravated the pathology and pathogenesis of IBV infection.     

Evaluation of rice–legume–rice cropping system on grain yield,nutrient uptake,nitrogen fixation,and chemical,physical, and biological properties of soil

To achieve higher yields and better soil quality under rice–legume–rice (RLR) rotation in a rainfed production system, we formulated integrated nutrient management (INM) comprised of Azospirillum (Azo), Rhizobium (Rh), and phosphate-solubilizing bacteria (PSB) with phosphate rock (PR), compost, and muriate of potash (MOP). Performance of bacterial bioinoculants was evaluated by determining grain yield, nitrogenase activity, uptake and balance of N, P, and Zn, changes in water stability and distribution of soil aggregates, soil organic C and pH, fungal/bacterial biomass C ratio, casting activities of earthworms, and bacterial community composition using denaturing gradient gel electrophoresis (DGGE) fingerprinting. The performance comparison was made against the prevailing farmers’ nutrient management practices [N/P₂O₅/K₂O at 40:20:20 kg ha⁻¹ for rice and 20:30:20 kg ha⁻¹ for legume as urea/single super-phosphate/MOP (urea/SSP/MOP)]. Cumulative grain yields of crops increased by 7–16% per RLR rotation and removal of N and P by six crops of 2 years rotation increased significantly (P < 0.05) in bacterial bioinoculants-based INM plots over that in compost alone or urea/SSP/MOP plots. Apparent loss of soil total N and P at 0–15 cm soil depth was minimum and apparent N gain at 15–30 cm depth was maximum in Azo/Rh plus PSB dual INM plots. Zinc uptake by rice crop and diethylenetriaminepentaacetate-extractable Zn content in soil increased significantly (P < 0.05) in bacterial bioinoculants-based INM plots compared to other nutrient management plots. Total organic C content in soil declined at 0–15 cm depth and increased at 15–30 cm depth in all nutrient management plots after a 2-year crop cycle; however, bacterial bioinoculants-based INM plots showed minimum loss and maximum gain of total organic C content in the corresponding soil depths. Water-stable aggregation and distribution of soil aggregates in 53–250- and 250–2,000 μm classes increased significantly (P < 0.05) in bacterial bioinoculants-based INM plots compared to other nutrient management plots. Fungal/bacterial biomass C ratio seems to be a more reliable indicator of C and N dynamics in acidic soils than total microbial biomass C. Compost alone or Azo/Rh plus PSB dual INM plots showed significantly (P < 0.05) higher numbers of earthworms’ casts compared to urea/SSP/MOP alone and bacterial bioinoculants with urea or SSP-applied plots. Hierarchical cluster analysis based on similarity matrix of DGGE profiles revealed changes in bacterial community composition in soils due to differences in nutrient management, and these changes were seen to occur according to the states of C and N dynamics in acidic soil under RLR rotation.     

Population genetic structure of <Emphasis Type="Italic">Rhizoctonia solani</Emphasis> AG1IA from rice field in North India

Rhizoctonia solani AG1IA is an important fungal pathogen causing significant yield and quality losses in rice production. However, little is known about the levels of genetic diversity and structure of this pathogen in North India. Out of 240 samples collected from different rice-growing regions of North India, 112 isolates were identified as R. solani AG1IA subgroups using species-specific primers. All 112 isolates were organized into four groups on the basis of percent disease index (PDI). The majority of the isolates were weakly virulent. Population genetic analysis was performed within and between populations using inter simple sequence repeat (ISSR) markers. A total of 8249 alleles were identified from the 112 isolates of R. solani AG1IA through analysis of the ten inter simple sequence repeat markers. All the ten ISSR markers were polymorphic. The average number of bands per primer was 7.3 which ranged in size from 250 to 1500 bp. Genetic structure of the isolates using inter simple sequence repeat primers showed high degree of polymorphism (PIC ≥0.81). The analysis of molecular variance (AMOVA) indicated that most of the genetic diversity occurred within populations (60%), while the variability among populations and among regions contributed 25 and 15%, respectively. Overall, the present study reveals that a large variation exists among rice-infecting isolates of R. solani AG1IA in North India. Fingerprinting of the isolates using ISSRs along with phenotypic characterization and virulence analysis will help epidemiological studies that can provide new insights into pathogen biology and disease spread.     

Combination of different marine algal extracts as biostimulant and biofungicide

This study investigates the efficiency of seaweed liquid fertilizer (SLF) prepared from combinations of different seaweeds (Sargassum polyphyllum, Turbinaria ornata, Gelidiopsis sp., Padina tetrastomatica, Gracilaria corticata) as a stimulant for the growth of Vigna radiata (Mung) as well as its antagonistic activity against fungal pathogens (Alternaria solani, Rhizoctonia solani., Sarocladium oryzae). 100% SLF was prepared, which was further diluted to 60%, 40%, and 20%. Seeds were soaked in four different concentrations of the SLF (20%, 40%, 60%, and 100%) for 12 hr and planted. After 60 d, the root and shoot length were increased by 14% and 16%, respectively, with SLF (100%). The carbohydrate and protein concentrations were also increased by 70% and 86%, respectively, at 100% SLF. The concentration of chlorophyll a, chlorophyll b, and carotenoids were found to be increased by 20%, 43%, and 28%, respectively, with 100% SLF. Further, the SLF (60% and 100%) successfully inhibited the growth of fungal pathogen A. solani but the other tested strains were found to be resistant. The present study indicates that 100% SLF concentration acts as both biostimulant and biofungicide for A. solani and thus, this SLF could be used as a potential alternative to the chemical fertilizers.