Smoke of Capsicum baccatumL. var. baccatum (Solanaceae) repels nymphs of Triatoma infestans(Klug) (Hemiptera: Reduviidae)

Control of the Chagas disease vector, Triatoma infestans(Klug) (Hemiptera: Reduviidae) with synthetic pesticides in Bolivia has become increasingly inefficient due to the development of resistance in the insects. In the Chaco region of Bolivia, guaraní populations have approached the problem by fumigating their houses with the smoke of native plants. Through interviews and field work with local guides, the main plant used by the guaraníes was collected and later identified as Capsicum baccatumL. var. baccatum(Solanaceae). In choice bioassays, filter papers exposed to the smoke of the plant repelled nymphs of T. infestans. Activity remained significant after storing the exposed filter papers for 9 days. Chemical analysis of smoke and literature data suggested that capsaicinoids present in the smoke were responsible for the repellent effect. The data presented provide a rationale for the use of C. baccatumvar. baccatumto control the Chagas vector bythe guaraní populations.

Molecular Epidemiology of Plasmodium species in Conflicted Federally Administered Tribal Area (FATA) Pakistan

Military conflicts have been significant obstacles in detecting and treating infectious disease diseases due to the diminished public health infrastructure, resulting in malaria endemicity. A variety of violent and destructive incidents were experienced by FATA (Federally Administered Tribal Areas). It was a struggle to pursue an epidemiological analysis due to continuing conflict and Talibanization. Clinical isolates were collected from Bajaur, Mohmand, Khyber, Orakzai agencies from May 2017 to May 2018. For Giemsa staining, full blood EDTA blood samples have been collected from symptomatic participants. Malaria-positive microscopy isolates were spotted on filter papers for future Plasmodial molecular detection by nested polymerase chain reaction (nPCR) of small subunit ribosomal ribonucleic acid (ssrRNA) genes specific primers. Since reconfirming the nPCR, a malariometric study of 762 patients found 679 positive malaria cases. Plasmodium vivax was 523 (77%), Plasmodium falciparum 121 (18%), 35 (5%) were with mixed-species infection (P. vivax plus P. falciparum), and 83 were declared negative by PCR. Among the five agencies of FATA, Khyber agency has the highest malaria incidence (19%) with followed by P. vivax (19%) and P. falciparum (4.1%). In contrast, Kurram has about (14%), including (10.8%) P. vivax and (2.7%) P. falciparum cases, the lowest malaria epidemiology. Surprisingly, no significant differences in the distribution of mixed-species infection among all five agencies. P. falciparum and P. vivax were two prevalent FATA malaria species in Pakistan’s war-torn area. To overcome this rising incidence of malaria, this study recommends that initiating malaria awareness campaigns in school should be supported by public health agencies and malaria-related education locally, targeting children and parents alike.

First Report of Lasiodiplodia pseudotheobromae Causing Collar Rot of Peanut in Shandong Province, China

In August 2019, a collar rot of peanut was observed in several fields in Qingdao, Shandong province, China. Disease survey was conducted in several peanut fields. Less than 5% plants exhibited various symptoms, including brown or black stem rot, pod rot, leaf chlorotic, wilted, and even dead. Symptomatic stems were cut into small pieces, surface disinfested with 70% ethanol for 1 min, 1% NaClO for 2 minutes, rinsed three times with sterile water, and dried on sterile filter papers. Pieces then were plated on potato dextrose agar (PDA) media and incubated at 25°C in darkness. Fungal cultures were initially white, then turned gray, and eventually turned black, and aerial hyphae were dense, fluffy. Conidia were ellipsoidal, initially hyaline, unicellular, 14.3 to 21.1 × 8.7 to 13.2 µm (n = 50), and mature conidia showed dark brown, with a central septum, and longitudinal stripes. Molecular identification was performed by sequencing ITS with ITS1/ITS4 (White et al., 1990) and beta tubulin gene with Bt2a/Bt2b (Glass and Donaldson, 1995) of a representative isolate ZHX9. ITS and beta tubulin regions (OK427342 and OK489788) of ZHX9 obtained 99.62 and 100% similar to L. pseudotheobromae (KF766193 and EU673111), respectively. Phylogenetic analysis was done using Neighbor-Joining (NJ) analysis based on those gene sequences. The microorganism we have isolated was identified as L. pseudotheobromae based on molecular analysis and morphological characteristics. For pathogenicity assay, twelve ten-days-old peanut (Zhonghua No.12) seedlings were each inoculated with one mycelial plug (8 mm in diameter) by placing the inoculum on the base of the stem. Twelve plants were each inoculated with a plug of non-colonized PDA as controls. Plants were incubated in a growth chamber (30°C in the day and 25°C at night, a 12-h photoperiod and 80% RH). Necrotic lesions were observed on stems of all inoculated seedlings 5 days after inoculation, whereas control plants remained asymptomatic, and L. peudotheobromae was consistently re-isolated from symptomatic stem. In Asia, peanut collar rot caused by L. teudotheobromae has been reported in India, Indonesia, North Vietnam (Nguyen, et al., 2006) and China (Guo, et al., 2014), but collar rot caused by L. pseudotheobromae has not been reported. To our knowledge, this is the first report of L. peudotheobromae causing collar rot on peanut in China. These results will provide crucial information for studying on epidemiology and management of this disease.

Influence of the Smoking and Spitting Behaviors in Public Smoking Areas on the Microbial Contamination and Spread

Objectives : The purpose of this study was to investigate the effect of spitting behavior on microbial public health in smoking areas. Through this investigation, bacterial concentration and spread of bacteria from the smoking areas were evaluated for the prospective public health and environment.Methods : The number of spitting and the spitting location in the smoking areas were measured visually by observers. For the microbial analysis, filter papers (5 × 5 cm) as sample collectors had been attached to the surface of the smoking areas for a given time. Using the collected filter paper, ATP bioluminescence measurement (RLU), colony forming unit (CFU) were measured. For the estimation of spitting behaviors in the smoking area on the spread microorganisms, saliva containing filters were selected from the areas, and fluorescent powder (0.01 g/cm2) was added to the papers. After 4 hours, the papers around the smoking areas were collected and the fluorescence activity was measured.Results and Discussion : The results showed that there was a clear relationship between the spitting behavior and the microbial activity in the smoking areas. In the case of sampling area A, the highest number of spitting was observed at the lunch hour (102 ± 16 times), and the RLU from the sample collected near the smoking area showed about 5,139 ± 1,267 RLU. When the RLU around the smoking area was measured, the lowest values were found at distance of 11 and 17 m from the cigarette bin (1,329 ± 148 and 1,204 ± 203 RLU), and it was confirmed that smoking and spitting did not occur at this sampling points. In the case of sampling area B, the highest number of spitting (45 ± 6 times) was observed at evening hour, and the RLU from the sample collected near the smoking area showed about 5,274 ± 1,297 RLU. The lowest value was in the sampling area B shown at 11 and 18 m (1,181 ± 243 RLU and 1,148 ± 168 RLU) from the cigarette bin, and it was also confirmed that smoking and spitting did not occur at this sampling points. The results indicate that the spitting during the smoking induced the increases of the microbial contamination of the public environment. The fluorescence powder diffusion analysis indicated that the saliva on the ground surface would enhance the spread of the microorganisms from the smoking area.Conclusions : Spitting in smoking areas can pose a high risk of microbial infection for smokers and non-smokers in smoking areas. Thus extensive and systematic attention related to the smoking manner must be paid to protect our public health.

Plasmodium falciparum Allelic Diversity: A Comparison of DNA Extraction from Isolates Collected on Rapid Diagnostic Tests (Rdts) and Filter Paper

Background: To perform molecular epidemiologic studies based on large cohorts, material such as RDTs or filter papers are useful for biological sample collection and extraction of RNA or DNA of good quality. Thus, we aimed to assess the quality of DNA extracted from malaria rapid diagnostic tests (RDTs) stored at various temperatures for the analysis of Plasmodium falciparum genetic diversity. Methods: Febrile patients benefitted from free malaria diagnosis using microscopy in a malaria sentinel site, at the Regional Hospital Estuaire-Melen, in Gabon, in 2015. P. falciparum isolates were collected onto one filter paper and 2 similar RDTs devices (Acon®) per patient. Nucleic acids were extracted with QiAmp Qiagen kit from paper and RDTs and the quality of the DNA was analyzed by msp1 gene amplification. Results: Msp1gene amplification was achieved in nucleic acids extracted from all filter papers and RDTs devices (n = 45, 100%). K1 alleles were detected in 93.3% (n = 42/45) of the samples and Mad20 alleles in 73.3% (n = 33/45). The number and the intensity of K1 and/or Mad20 fragments were comparable according to the sample collection material and the storage conditions (room temperature vs -20°C) of the samples. The size of the fragments indicating allelic diversity was comparable in 80% (n=36) of the samples. Conclusion: These data show that RDTs are a valuable source of DNA for malaria parasite genetic polymorphism analysis. Storage conditions of the devices did not influence the quality of DNA extracted from RDTs device, although some alleles may be missed.

Improvement of Bovine Pestiviral diagnosis by the development of a cost-effective method for detecting viral RNA in fresh specimens and samples spotted in filter papers.

Bovine pestiviruses are the causative agents of Bovine viral diarrhea, a disease that generates severe economic losses in cattle. The aim of this study was to improve its diagnosis by means of developing a RT-qPCR to detect bovine pestiviruses A, B and H; and to set a protocol for collecting, shipping and conserving bovine pestiviral RNA in filter papers. The developed RT-qPCR showed high sensitivity in detecting these viruses in different matrixes: viral stocks, semen and serum samples. Regarding the possibility of using the technique to test serum pools, it was possible to identify a positive serum sample within a pool containing 30 sera. In addition to evaluating the qPCR from fresh samples, the use of filter papers to sow bovine samples was analyzed. The sampling method in two different filter papers using bovine blood drops was a useful alternative for diagnosis purposes and allowed the preservation of pestiviral RNA up to 12 months under refrigeration.

The Residual Efficacy of SumiShield™ 50WG and K-Othrine® WG250 IRS Formulations Applied to Different Building Materials Against Anopheles and Aedes Mosquitoes

Insecticides with novel modes of action are required to complement the pyrethroids currently relied upon for controlling malaria vectors. One example of this is the neonicotinoid clothianidin, which is found in SumiShield™ 50WG used in indoor residual spraying (IRS). In a preliminary experiment, mortality in insecticide susceptible and resistant An. gambiae adults exposed to SumiShield™ 50WG-treated filter papers reached 80% by 3-days post-exposure and 100% by 6-days post-exposure. Next, cement, wood, and mud tiles were treated with SumiShield™ 50WG or K-Othrine® WG250 (deltamethrin IRS formulation) and insecticide resistant and susceptible Anopheles and Aedes were exposed to these surfaces periodically for up to 18-months. Pyrethroid resistant Cx. quinquefasciatus were also exposed at 9 months. Between exposures tiles were stored in heat and relative humidity conditions reflecting those found in the field. On these surfaces, SumiShield™ 50WG was effective at killing both susceptible and resistant An. gambiae for 18 months post-treatment, while mortality amongst the resistant strains when exposed to deltamethrin (K-Othrine® WG250) IRS was not above that of the negative control. Greater efficacy of SumiShield™ 50WG was also demonstrated against insecticide resistant strains of An. funestus compared to deltamethrin, though the potency was lower when compared with An. gambiae. In general, a higher efficacy of SumiShield™ 50WG was observed on cement and mud compared to wood. SumiShield™ 50WG demonstrated poor residual activity against Aedes aegypti and Culex quinquefasciatus. Overall, the results suggest SumiShield™ 50WG is well suited for malaria control.

Insecticidal Activities of Leaf Methanol Extract of Clerodendrum polycephalum Baker against Aedes aegypti, Anopheles gambiae and Culex quinquefasciatus

Aim: In a view to determining the capacity of Clerodendrum polycephalum to control mosquitoes, the methanol extract of the leaf was investigated for insecticidal activities using three species of mosquitoes, Aedes aegypti, Anopheles gambiae and Culex quinquefasciatus. Methodology: The leaves of C. polycephalum were collected, dried and extracted with methanol. Mosquito larvae were exposed to different concentrations for 24/48h. and sublethal concentrations (L25, 50, 75) of the extract to determine larvicidal activity and monitor growth and development respectively. Twenty (20) blood-fed female Aedes aegyti mosquitoes were allowed to lay eggs on treated filter papers for antioviposition bioassay. The crude extract was separated into N-hexane, Dichloromethane, ethyl-acetate, ethanol fractions using Vacuum Liquid Chromatography to determine the active fraction. Results: Results showed that larval mortalities were in the order C. quiquefasciatus>A. gambiae>A. aegypti with effective concentration ranging from 250 – 8000 ppm. Mortalities at 48hr were significantly different (p<0.05) from mortalities at 24h. Of the four fractions obtained, ethanol fraction gave the highest larval mortality of 100% at 2000 ppm. Larval duration for all the three species of mosquitoes exposed to the crude extract varied between 2.25±0.5 and 3.25±0.5 days. As Antioviposition Index increased from 46.4 - 89.9, percent hatching of mosquito eggs deceased from 87.25 – 67.5% with increasing concentration. Conclusion: The extract of C. polycephalum was found to contain insecticidal compounds which are soluble in polar solvent. The plant could be exploited in mosquito control programme.

High-Temperature Stable Specific Enzyme for Guar Polymer Based Fracturing Fluid Degradation

Hydraulic fracturing, commonly referred to as fracking, is a widely used technology to enhance the productivity of low-perm reservoirs and the aqueous-based fracturing fluids use guar as the rheology builder. Residual polymer layer over the fractured surface results in a reduced matrix to fracture permeability, causing reduced well productivity. This research aims to develop a specialized mannanase enzyme and evaluate its efficiency in degrading linear and cross-linked guar polymer gel as a function of time, temperature, and breaker concentration, to enhance the effectiveness of the fracturing process and yielding higher production. The study begins with developing high-temperature stable mannanase using "protein engineering" tools to minimize denaturation at high temperatures and the underlying formation chemistry, followed by optimization of polymer, crosslinker, and breaker concentration through the measurement of rheological properties at moderate to high temperature. Initial studies were conducted using HT-HP filter press and filter papers as porous media for visual inspection of polymer cake dissolution efficiency. Final conclusions were drawn from the simulated coreflooding studies, wherein the injection and production return permeabilities were investigated on post-fracture and enzyme-treated cores, where the breaker was mixed with the frac fluid applied once the frac fluid is in place. The thermal stability of the enzyme breaker vis-à-vis viscosity reduction and degradation pattern of linear and cross-linked gel observed from the break test showed that the enzyme is stable up to 250 °F and can reduce viscosity by more than 1800 cp (99% breaking ability).

New Evidence of Model Crop Brassica napus L. in Soil Clean-Up: Comparison of Tolerance and Accumulation of Lead and Cadmium

The potential of the model crop Brassica napus L. (rapeseed) for the phytoremediation of soils polluted with metals was investigated at laboratory scale. The first step consists in the evaluation of the seed germination and growth of the Brassica napus L. plant in a controlled environment, followed by the determination of the photosynthetic pigments content represented by chlorophyll a and b and carotenoids. The degree of metal accumulation in rapeseed has been evaluated by the bioaccumulation factor (BAC), the bioconcentration factor (BCF) and the translocation factor (TF). Phytotoxicity tests were performed in Petri dishes with filter papers moistened with metal solutions in the range of 0 to 300 mg/L Pb(II) or Cd(II). At the highest concentration of the lead and cadmium treatments (300 mg/L), B. napus L. showed the lowest germination degree (56.67% and 43.33%, respectively). According to Tukey test results, Pb(II) concentrations of up to 300 mg/L do not significantly affect the length of the hypocotyls, whereas, in the case of Cd(II), the mean of the radicle and hypocotyl lengths of the seedlings are significantly affected compared to the mean of the control. In soil pot experiments, important changes have been obtained in the pigment content, especially in the case of cadmium. For both metals and for each treatment (100 to 1500 mg/kg Pb(II) and 1 to 30 mg/kg Cd(II)), a TF < 1 indicates an ineffective metal transfer from root to shoot. Finally, rapeseed can be considered a tolerant plant and a suitable candidate for Pb(II) and Cd(II) accumulation and for the phytostabilization of contaminated soil under the experimental conditions adopted in the present study.