Identification of immunoreactive polypeptides of Babesia equi parasite during immunization

This study was carried out to identify immunoreactive polypeptides in Babesia equi merozoite antigen. Three fractions of killed B. equi merozoite antigen viz.; whole merozoite (WM), cell membrane (CM) and high speed supernatant (HSS) antigens were prepared from the parasite infected erythrocytes. These antigenic preparations along with ghost antigen from non-infected erythrocytes were fractionated on 12% sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) and immunoblotted with sera showing high antibody titres. On SDS-PAGE, 16 polypeptides with molecular weight (Mr) in the range of 112-17kDa were obtained from the WM and CM antigens. But only six polypeptides were detected (96.5-28kDa) in the HSS antigen. On immunoblotting with high titred serum collected from donkeys following two immunizations with a killed B. equi merozoite immunogen, 11 polypeptides were observed in the WM and CM antigens (Mr 112-18kDa). Of these, four polypeptides (Mr 112, 45, 33 and 18kDa) were identified as most immunoreactive. Besides these, a 28kDa was observed as strong immunoreactive protein in WM and CM antigens. The HSS antigen showed only six polypeptides and one peptide (28kDa) was identified as immunoreactive. When high titred serum collected from immunized donkeys following challenge with B. equi infected blood and was used for immunoblotting, the protein profile of WM and CM antigens remained the same. However, three additional polypeptides (Mr 81, 54.5 and 39kDa) were detected in HSS antigen.     

Temporal changes in endogenous estrogens and expression of behaviors associated with estrus during the periovulaory period in Murrah buffaloes (<Emphasis Type="Italic">Bubalus bubalis</Emphasis>)

The objective of this study were (1) to establish the duration of behavioral estrus signs and timing of ovulation in Murrah buffaloes (n = 10) and (2) to determine relationship between behavioral estrus signs with change in plasma estrogen concentrations. Estrus and its behavioral signs were detected at hourly intervals by visual observations, per recta examination of genitalia and bull parading four times in a day for 30 minutes each. Among the behavioral signs of estrus, swollen vulva (80%) was the best indicator of estrus followed by excitement (70%). Among the duration of behavioral estrus signs the first and longest duration of estrus signs was swollen vulva which was seen upto 19.8 ± 0.8 h after onset of estrus. The mean total duration of estrus symptoms from appearance to disappearance of all the behavioral estrus symptoms was 23.5 ± 1.7 h. All the behavioral estrus symptoms were observed during the period of estrogen surge. Endocrine profile during the periestrus period showed that the mean peak concentrations of total estrogen 23.9 ± 3.9 pg/ml occurred at 8.8 ± 1.7 h after onset of estrus. The average number of estrus symptoms observed per animal during onset of spontaneous estrus was 5.7. Ovulation occurred after 37.4 ± 1.7 h after onset of estrus and 13.4 ± 1.0 h after end of total estrogen surge respectively. In conclusion, our results suggest that all signs of behavioral estrus occurred during the preovulatory rise in estrogens. The first sign of estrus to be observed was a swollen vulva and this symptom persisted the longest.     

Experimental avian paramyxovirus serotype-3 infection in chickens and turkeys

Avian paramyxoviruses (APMV) are divided into nine serotypes. Newcastle disease virus (APMV-1) is the most extensively characterized, while relatively little information is available for the other APMV serotypes. In the present study, we examined the pathogenicity of two divergent strains of APMV-3, Netherlands and Wisconsin, in (i) 9-day-old embryonated chicken eggs, (ii) 1-day-old specific pathogen free (SPF) chicks and turkeys, and (iii) 2-week-old SPF chickens and turkeys. The mean death time in 9-day-old embryonated chicken eggs was 112 h for APMV-3 strain Netherlands and > 168 h for strain Wisconsin. The intracerebral pathogenicity index in 1-day-old chicks for strain Netherlands was 0.39 and for strain Wisconsin was zero. Thus, both strains are lentogenic. Both the strains replicated well in brain tissue when inoculated intracerebrally in 1-day-old SPF chicks, but without causing death. Mild respiratory disease signs were observed in 1-day-old chickens and turkeys when inoculated through oculonasal route with either strain. There were no overt signs of illness in 2-weeks-old chickens and turkeys by either strain, although all the birds seroconverted after infection. The viruses were isolated predominantly from brain, lungs, spleens, trachea, pancreas and kidney. Immunohistochemistry studies also showed the presence of large amount of viral antigens in both epithelial and sub-epithelial lining of respiratory and alimentary tracts. Our result suggests systemic spread of APMV-3 even though the viral fusion glycoprotein does not contain the canonical furin proteases cleavage site. Furthermore, there was little or no disease despite systemic viral spread and abundant viral replication in all the tissues tested.     

Comparative study of conventional and artificial neural network-based ETo estimation models

Accurate estimation of reference crop evapotranspiration (ETo) is required for several hydrological studies and thus, in the past, a number of ETo estimation methods have been developed with different degree of complexity and data requirement. The present study was carried out to develop artificial neural network (ANN) based reference crop evapotranspiration models corresponding to the ASCE’s best ranking conventional ETo estimation methods (Jensen et al. ASCE Manual and Rep. on Engrg. Pract. no. 70, 1990). Among the radiation methods, FAO-24 radiation (or Rad) method for arid and Turc method for humid region, and among the temperature methods, FAO-24 Blaney–Criddle (or BC) method were studied. The ANN architectures corresponding to the above three less data-intensive methods were developed for four CIMIS (California Irrigation Management Information System) stations, namely, Davis, Castroville, Mulberry, and West Side Field station. The comprehensive ANN architecture developed by Kumar et al. (J Irrig Drain Eng 128(4):224–233, 2002) corresponding to Penman–Monteith (PM) ETo for Davis was also tried for the other three stations. Daily meteorological data for a period of more than 10 years (01 January 1990 to 30 June 2000) were collected from these stations and were used to train, test, and validate the ANN models. Two learning schemes, namely, standard back-propagation with learning rate of 0.2 and standard back-propagation with momentum having learning rate of 0.2 and momentum term of 0.95 were considered. ETo estimation performance of the ANN models was compared with the FAO-56 PM method. It was found that the ANN models gave better closeness to FAO-56 PM ETo than the best ranking method in each category (radiation and temperature). Thus these models can be used for ETo estimation in agreement with climatic data availability, when not all required climatic variables are observed.     

Larval and pupal toxicity effects of Plectranthus amboinicus, Bacillus sphaericus and predatory copepods for the control of the dengue vector, Aedes aegypti

The efficacy of copepod Mesocyclops aspericornis (Daday) combined with the larvicide Bacillus sphaericus (Bs) and a plant extract of Plectranthus amboinicus leaf abstract (PALE), used jointly and singly, was studied against Aedes aegypti in the laboratory. P. amboinicus leaf extract of 20, 40, 60, 80 and 100 ppm caused significant mortality against Ae. aegypti larvae. The LC₅₀ and LC₉₀ values for I to IV instars larvae and pupae were 26.12, 35.36, 45.76, 52.32 and 63.82 ppm, respectively. The LC₉₀ values for I to IV instars larvae and pupae were 82.53, 92.65, 108.06, 119.47 and 131.71 ppm, respectively. Under laboratory conditions, copepods treatment produced 7.9% predatory efficiency against 1st instar larvae of Ae. aegypti, at a copepod:larvae ratio of 1:10. When copepod treatment was combined with PALE this was increased to 8.7. The treatment of copepods combined with Bs and PALE yielded a better and more sustainable result (9.6%) than the agents used individually. This predation efficiency may be caused by detrimental effects of the P. amboinicus active principle compound (carvacrol) on the mosquito larvae. Our results suggest that the combined application of microbial insecticide (Bs), copepods and P. amboinicus leaf extract may be used to control Aedes populations.     

Biological response of chilli thrips, Scirtothrips dorsalis Hood (Thysanoptera: Thripidae), to various regimes of chemical and biorational insecticides

The chilli thrips Scirtothrips dorsalis (Hood) (Thysanoptera: Thripidae), a new invasive pest in the USA, is an economically important pest of certain vegetable, ornamental and fruit crops in southern and eastern Asia, Oceania and parts of Africa. These crops cannot be protected from the pest without resorting to the use of chemical insecticides. In order to forestall or delay the development of insecticide resistance in S. dorsalis, we continued our focus on the discovery of insecticides with different modes of action for rotational use. In this study we evaluated candidate insecticides to control S. dorsalis on ‘Jalapeno’ pepper, Capsicum annuum L.; these materials belong to different IRAC mode of action classes as follows: (i) 4A – neonicotinoids, i.e., imidacloprid, thiamethoxam and dinotefuran, (ii) 5 – spinosyns, i.e., spinosad and spinetoram, (iii) 3A – pyrethroids, i.e., β-cyfluthrin, esfenvalerate, ζ-cypermethrin and λ-cyhalothrin and (iv) 8D – borax mixed together with orange oil and detergents in the TriCon^® formulation. In addition we evaluated the entomopathogenic fungus, Beauveria bassiana (Botanigard^®) alone and in combination with the borax formulation at ½ of their usual rates of application. Each of the 3 neonicotinoid insecticides when applied either as a single foliar spray or as a soil drench significantly suppressed both adults and larvae for at least 10 days; indeed imidacloprid did so for 15 days. Dinotefuran was more effective as a foliar spray than as a soil drench. Spinosyns applied as a single foliar spray, significantly suppressed both adults and larvae through 15 days after treatment (DAT). None of the 4 pyrethroids provided significant suppression of either adults or larvae. The borax formulation suppressed adults and larvae through 10 DAT. B. bassiana significantly suppressed only the larvae at 5 DAT and not at 10 DAT. This study brings the number of insecticides known to be effective against S. dorsalis to 10 and these belong to 7 different modes of action classes. The use of such insecticides in rotation belonging to different classes will help delay the development of insecticide resistance in S. dorsalis.     

OsSUV3 transgenic rice maintains higher endogenous levels of plant hormones that mitigates adverse effects of salinity and sustains crop productivity

Background

The SUV3 (suppressor of Var 3) gene encodes a DNA and RNA helicase, which is localized in the mitochondria. Plant SUV3 has not yet been characterized in detail. However, the Arabidopsis ortholog of SUV3 (AT4G14790) has been shown to be involved in embryo sac development. Previously, we have reported that rice SUV3 functions as DNA and RNA helicase and provides salinity stress tolerance by maintaining photosynthesis and antioxidant machinery. Here, we report further analysis of the transgenic OsSUV3 rice plants under salt stress.

Findings

The transgenic OsSUV3 overexpressing rice T₁ lines showed significantly higher endogenous content of plant hormones viz., gibberellic acid (GA₃), zeatin (Z) and indole-3-acetic acid (IAA) in leaf, stem and root as compared to wild-type (WT), vector control (VC) and antisense (AS) plants under salt (200 mM NaCl) stress condition. A similar trend of endogenous plant hormones profile was also reflected in the T₂ generation of OsSUV3 transgenic rice under defined parameters and stress condition.

Conclusions

In response to stress, OsSUV3 rice plants maintained plant hormone levels that regulate the expression of several stress-induced genes and reduce adverse effects of salt on plant growth and development and therefore sustains crop productivity.     

Integrated effect of population and weed management regimes on weed dynamics,performance, and productivity of basmati rice (<Emphasis Type="Italic">Oryza sativa</Emphasis> L.)

Weeds caused serious problem on yield reduction of basmati rice worldwide. Losses caused by weeds varied from one country to another, depending on the presence of dominant weeds and the control methods practiced by farmers; therefore, suitable plant population and weed management practices should be adopted. Keeping these in mind, a field experiment was carried out during kharif seasons of 2009 and 2010 at crop Research Centre of SVPUA&T, Meerut, India comprising 4 planting geometries, viz. 20, 30, 40, and 50 hills m^?2 as main plot factor, and 5 weed management practices (Butachlor @ 1.0 kg ha^?1, Butachlor @ 1.0 kg ha^?1 fb (followed by) one hand weeding, Butachlor @ 1.0 kg ha^?1 fb Almix @ 4 g ha^?1, two hand weedings and weedy check) in a split plot design with 3 replications. Experimental results revealed that plant population of 50 hills m^?2 proved superior over that of 20 hills m^?2 in respect of weed density, weed dry weight, number of tillers m^?2, yield attributes, grain, straw, and biological yields. The maximum grain yield (29.00 and 31.00 q ha^?1) and straw yield (51.30 and 52.50 q ha^?1) were recorded in 50 hills m^?2 followed by 40 hills m^?2 during 2009 and 2010, respectively. In respect of nitrogen, phosphorus, and potassium removal, a reverse trend was observed: the highest in 20 hills m^?2 followed by 30, 40, and 50 hills m^?2. As far as the weed management practices are concerned, both chemical and mechanical methods of weed control were found superior over weedy check. The lowest weed density, dry weight, and highest weed control efficiency, maximum length of panicle^?1, number of panicle (m²), and 1000-grain weight and grain yield of 30.40 and 32.60 q ha^?1 were recorded with two hand weedings which was at par with Butachlor @ 1.0 kg ha^?1 fb one hand weeding over rest of the weed management practices.     

Biochemical and spectroscopic characterization of morning glory peroxidase from an invasive and hallucinogenic plant weed Ipomoea carnea

A novel heme peroxidase MGP from the latex of Ipomoea carnea subsp. fistulosa (morning glory) belonging to the Convolvulaceae family was purified to homogeneity using ammonium sulfate precipitation, anion exchange, hydrophobic interaction, and gel filtration chromatography. The enzyme is glycosylated and has a molecular mass of 42.06 kDa (MALDI-TOF) and an isoelectric point of pH 4.3. The enzyme has high yield, broad substrate specificity, and a high stability toward pH, temperature, chaotrophs, and organic solvents. The extinction coefficient (epsilon 280 (1%)) of the enzyme was estimated as 20.56 and it consists of 13 tryptophan, 9 tyrosine, and 8 cysteine residues forming 4 disulfide bridges. There is significant effect of inhibitors targeting S-S bridges (mercaptoethanol, l-cysteine, glutathione), as well as of inhibitors targeting heme (sodium azide and hydroxylamine) on peroxidase activity, whereas inhibition was not observed with ethylmaleinimide due to the absence of reduced cysteine in the enzyme. Polyclonal antibodies against the enzyme have been raised in rabbit, and immunodiffusion suggests that the antigenic determinants of MGP are unique. The N-terminal sequence of MGP (D-E-A-C-I-F-S-A-V-K-E-V-V-D-A) exhibited considerable similarity to the sequence of other known plant peroxidases. Spectroscopic studies (absorbance, fluorescence, and circular dichroism) reveal that MGP has secondary structural features with alpha/beta type with approximately 20% alpha-helicity.