Isolation and molecular characterization of entomopathogenic nematode,Heterorhabditis sp. from an arable land in Nigeria

The occurrence of entomopathogenic nematodes(EPNs) in arable soil samples from Nigeria was investigated using Baermann extraction tray and insect-bait(White's trap) techniques. Isolates were tested for infectivity using the larvae of Galleria mellonella(greater moth) and Tenebrio molitor(mealworm). The study revealed a new species of Heterorhabditis(MT371593) in soil samples that were randomly collected from an arable farmland cultivated with cassava TMS-30572 at the Teaching and Research Farm of Landmark University, Nigeria. Amplification of the internal transcribed spacer region(ITS) of the ribosomal DNA produced a nucleotide sequence of 933 base pairs(bp). A BLASTN search of Gen Bank showed that the sequence of the Nigerian isolate is identical at 99% similarity to that of Heterorhabditis sp. from Thailand. Infectivity test of the isolate showed 100% mortality against T. molitor larvae within 48 h of exposure while only 80% mortality was recorded for G. mellonella after 1 week of exposure. This is the first account of Heterorhabditis sp. in Nigeria. The varying degrees of infectivity against mealworm and greater moth observed in this study proved that the Nigerian isolate of Heterorhabditis sp. could potentially be an attractive option in the management of insect pests of cash crops.     

Length‐based assessment of larval lamprey population structure at differing spatial scales

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Bioaccumulation of cadmium and its biochemical effect on selected tissues of the catfish (<Emphasis Type="Italic">Clarias gariepinus</Emphasis>)

The present study examines the pattern of accumulation of cadmium (Cd) and its biochemical effects on selected tissues of a variety of African catfish (Clarias gariepinus), after exposure to various doses of Cd. The results obtained indicate that at the end of 21 days of exposure, the total tissue organ cadmium concentration followed the pattern kidney > gill > liver > muscle for each of the exposure concentrations. The levels of Cd in these organs were higher than those in ambient water. Moreover, while the rate of uptake of Cd increased with time in the kidney, liver and muscle, it decreased in the gill. Superoxide dismutase (SOD) was significantly elevated only in the kidney of catfish treated with 0.2 and 0.4 ppm of Cd for 7 days compared with the control. Conversely, gill SOD was significantly decreased in the same concentrations of Cd-treated catfish relative to the control. Statistically similar levels of SOD were observed in the liver, brain and muscle with all the treatments after the same duration of treatment. In the fish exposed for 21 days, SOD activity was significantly decreased in the kidney with a corresponding increase in lipid peroxidation (LPO), but it manifested only with the 0.2- and 0.4-ppm Cd treatment relative to the control. In the liver, however, Cd exposure significantly increased SOD in the 0.2- and 0.4-ppm treatments of the same duration. In conclusion, the present study indicates that the accumulation of Cd and its effect on SOD and LPO in C. gariepinus is dependent on concentration, tissue and time     

Effects of Vermicompost and Vermiwash on Plant,Phenolic Content,and Anti-oxidant Activity of Mexican Pepperleaf (Piper auritum Kunth) Cultivated in Phosphate Rock Potting Media

The aim of this investigation was to determine the effect of vermicompost, vermiwash, and phosphate rock on plant, total phenols, flavonoids, and anti-oxidant activity in Piper auritum Kunth leaves. P. auritum plants were obtained from cuttings and were planted according to the Box-Behnken experimental design with three repetitions at the central point. The factors and levels were vermicompost (10, 20, and 30 g plant^?1), vermiwash (5, 10, and 15 mL plant^?1), and phosphate rock (1, 2, and 3 g plant^?1). Plant growth parameters (plant height, stem diameter, leaves number) and chlorophyll content were measured 1 month after treatment applications. Total phenols, total flavonoids, and 1,1-diphenyl-2-picryl-hydrazyl radical scavenging activity was measured after 4 months. Vermicompost, vermiwash, and phosphate rock had no statistically significant effect on plant growth. Plant height, stem diameter, leaves number, chlorophyll, innermost number, fresh weight stem, fresh weight leaves, fresh weight root, dry weight stem, dry weight leaves, and dry weight root were not different among treatments. Total phenolic compounds were statistically affected for both vermiwash and phosphoric rock (p < 0.05) and the anti-oxidant activity decreased by vermicompost addition. The application of 15 mL plant^?1 vermiwash, 1 g phosphate rock, and 20 g vermicompost plant^?1 increased the total phenol content.     

Susceptibility of the Mexican bean beetle <Emphasis Type="Italic">Epilachna varivestis</Emphasis> Mulsant (Coleoptera: Coccinellidae) to endemic isolates of <Emphasis Type="Italic">Beauveria bassiana</Emphasis> (Balsamo) Vuillemin (Hypocreales: Clavicipitaceae)

Entomopathogenic fungi represent excellent candidates for biological insecticides. Among the many entomopathogenic fungi, Beauveria bassiana (Balsamo) Vuillemin (Hypocreales: Clavicipitaceae) is one of the most promising species. Here, we report the results of infection studies performed using four B. bassiana native isolates (Bb37, Bb38, Bb40, and Bb45) compared with the commercial product Mycotrol^® (strain GHA) on adult Mexican bean beetle Epilachna varivestis Mulsant (Coleoptera: Coccinellidae), one of the most important pests of bean production in Mexico. First, single concentration (1 × 10⁸ conidia mL^?1) assays were carried out using all isolates applied using an immersion method. Examinations at 10 days after inoculation indicated that isolates Bb37, Bb38, and Bb40 significantly reduced adult survival, causing 60–75% mortality compared to the commercial strain GHA (33%) and control (29%). Second, using two selected isolates (Bb37 and Bb40), the mean lethal concentration values (LC₅₀) were estimated for third instar larvae and adult E. varivestis. The LC₅₀ values for Bb37 and Bb40 isolates decreased 5.26-fold and 3.19-fold, respectively, in third instars compared with adults. However, such difference for Bb40 isolate was not significant. Finally, an experiment, to compare the median survival time (MST) values between third instar and adult E. varivestis, was conducted using Bb37 and Bb40 at 1 × 10⁹ conidia mL^?1. MST values were significantly reduced against larvae (87 and 100 h) compared with adults (130 and 134 h) by Bb37 and Bb40, respectively. We conclude that native isolates of B. bassiana represent an important alternative for the control of E. varivestis. However, to demonstrate its effectiveness under field conditions, detailed studies are needed.     

Learning by doing: a participatory methodology for systematization of experiments with agroforestry systems, with an example of its application

Participatory research methods have helped scientists to understand how farmers experiment and to seek partnerships with farmers in developing technologies with enhanced relevance and adoption. This paper reports on the development of a participatory methodology to systematize long-term experimentation with agroforestry systems carried out in a hotspot of biodiversity by non-governmental organizations and local farmers. A methodological guide for systematization and techniques used for Participatory Rural Appraisal formed the basis of our work. We propose an analytical framework that recognizes systems of reflexive and learning interactions, in order to make the learned lessons explicit. At the process level, the main lessons and recommendations are as follows. It is important to establish partnerships to conduct innovative and complex experimentation with agroforest. Participatory systematization allows us to improve the methodological aspects of design, implementation and management of on-farm participatory experimentation. It also serves to synthesize the main findings and to extract lessons from agroforestry systems experiments. It fosters the technical improvement of agroforestry systems. It creates possibilities for reflection on agroforestry systems by farmers, extensionists and researchers, as well as their learning with respect to management of such systems. The findings are placed in the context of current theory on participatory experimentation in agriculture. Extractive and interactive approaches help to produce rich insights of mutual interest through collaboration by identifying local, regional and global convergences, complementarities, and conflicts of interest; which affect the advance of new eco-friendly technologies, to both improve the livelihoods and to reverse biodiversity loss and environmental degradation.     

Spatio-temporal dynamics of gross rainfall partitioning and nutrient fluxes in shaded-cocoa (<Emphasis Type="Italic">Theobroma cocoa</Emphasis>) systems in a tropical semi-deciduous forest

Land-use change from forest to cocoa agroforestry and other tree-based farming systems alters the structure of forest stands and influences the magnitude of canopy water fluxes and subsequent bio-element inputs to the forest floor. The partitioning of incident rainfall (IR) into throughfall (TF), stemflow (SF) and canopy interception loss (IL_C) and their associated nutrient element concentrations and fluxes was examined along a replicated chrono-sequence: forest, 3, 15 and 30-year-old smallholder shaded-cocoa systems in Ashanti Region, Ghana. Mean annual precipitation during the 2-year observational period (2007 and 2008) was 1376.2 ± 93.8 mm. TF contributed between 76.5–90.4%, and SF between 1.4–1.7% of the annual IR to the forest floor. There were significant differences in IR, TF and SF chemistry. While TF and SF were enriched in phosphorus (1.33–5.67-fold), potassium (1.1–5.69 fold), calcium (1.35–2.65 fold) and magnesium (1.4–2.68 fold) relative to IR, total N (NH₄ ⁺+NO₃ ^?) declined (0.5–0.91) of IR values in TF and SF in forest and shaded cocoa systems. Incident rainfall was significantly more acidic than TF and SF in both forest and shaded-cocoa systems. Mean annual total N, P, K, Ca and Mg inputs to the forest floor through IR were 5.7, 0.14, 13.6, 9.43 and 5.6 kg ha^?1year^?1 respectively. Though an important source of available nutrients for plant growth, incident rainfall provides only a small percentage of the annual nutrient requirements. With declining soil fertility and pervasive low cocoa yields, possible effects of the reported nutrient fluxes on nutrient budgets in cocoa systems merit further investigation. Against the background of increased TF and decreased IL_C following forest conversion to shaded-cocoa, it is also recommended that more studies be carried out on rainfall partitioning and its impact on ground water recharge as a way of establishing its influence on the availability of moisture for agriculture in these systems.     

Multiply-primed rolling-circle amplification (MPRCA) of PCV2 genomes: Applications on detection, sequencing and virus isolation

Multiply-primed rolling-circle amplification (MPRCA) was used to amplify porcine circovirus type 2 (PCV2) genomes isolated from tissues of pigs with signs of post-weaning multisystemic wasting syndrome (PMWS). Two of the amplified PCV2 genomes were cloned in prokaryotic plasmids and sequenced. Both were nearly identical (1767 nt) except for one silent substitution in the region coding for the capsid protein (ORF2). In addition, they showed high nucleotide sequence similarity with PCV2 isolates from others countries (93–99%). To investigate whether the MPRCA amplified PCV2 genomes could be used to produce infectious virus, the cloned genomes were isolated from the plasmids, recircularized and used for transfection in PK-15 cells. This procedure led to the production of infectious virus to titres up to 10^5.55 TCID₅₀/mL. It was concluded that MPRCA is a useful tool to amplify PCV2 genomes aiming at sequencing and virus isolation strategies, where particularly useful is the fact that it allows straightforward construction of PCV2 infectious clones from amplified genomes. However, it was less sensitive than PCR for diagnostic purposes.     

Tick salivary gland extract-activated transmission of Borrelia afzelii spirochaetes

Saliva-activated transmission of Borrelia afzelii Canica, Nato, du Merle, Mazie, Baranton et Postic, 1993 was demonstrated using salivary gland extract (SGE) from Ixodes ricinus (L., 1758) ticks and C3H mice. Injection of Borrelia spirochaetes together with SGE increased the level of bacteraemia and accelerated the appearance of bacteria in the urinary bladder, compared with the injection of spirochaetes alone. More I. ricinus nymphs became infected when feeding on mice inoculated with B. afzelii plus SGE. Analysis of cytokines produced by cells of draining lymph nodes from SGE-treated mice showed a suppression of proinflammatory cytokines IFN-gamma, IL-6 and GM-CSF following a transient upregulation in comparison with the control mice infected without SGE.     

Swine viscera preservation in hypersaturated salt solution after alcohol fixation as a preparation method for educational purposes

The use of live animals for educational purposes is an old practice that is still employed in teaching and research institutions. However, there are several objections to this practice, whether for ethical or humanitarian reasons. Surgical techniques teaching using anatomical pieces and/or preserved cadavers promotes greater learning efficiency, provides exercise repetition and increases the confidence and satisfaction of the students when compared to the use of live animals. The current work aimed to analyse the feasibility of using fresh swine urinary bladder and small intestines (jejunum), obtained from slaughterhouses, fixed in 99.8% ethyl alcohol (EA) and preserved in sodium chloride hypersaturated solution (SCHS) at 30%, for 7, 14 and 21 days, as an alternative method for surgical skills training (SST). Swine viscera, fixed in EA and preserved in SCHS, presented a realistic appearance, absence of odour and maintained the viable morphological characteristics during the performance of the operative techniques. Preservation solutions had low cost, were easy to acquire and did not offers risks to human health. Therefore, urinary bladders and small intestines fixed in 99.8% EA for 30 days and maintained in 30% SCHS at different periods were demonstrated as a good viable option as a preservation method for surgical skills training.