Uganda is the only country in sub-Saharan Africa whose onchocerciasis elimination programme extensively uses vector control and biannual treatment with ivermectin. The purpose of this study was to assess the impact of combined strategies on interrupting onchocerciasis transmission in the Kashoya-Kitomi focus. Mass Drug Administration annually (13 years) followed by biannual treatments (6 years) and ground larviciding (36 cycles in 3 years) with temephos (Abate®, EC500) against Simulium neavei were conducted. Routine fly catches were conducted for over seven years in six catching sites and freshwater crabs Potamonautes aloysiisabaudiae were examined for immature stages of Simulium neavei. Epidemiological assessments by skin snip were performed in 2004 and 2013. Collection of dry blood spots (DBS) from children <10 years for IgG4 antibodies analysis were done in 2010 and 2013. Treatment coverage with ivermectin improved with introduction of biannual treatment strategy. Microfilaria prevalence reduced from 85% in 1991 to 62% in 2004; and to only 0.5% in 2013. Crab infestation reduced from 59% in 2007 to 0% in 2013 following ground larviciding. Comparison of total fly catches before and after ground larviciding revealed a drop from 5334 flies in 2007 to 0 flies in 2009. Serological assays conducted among 1,362 children in 2010 revealed 11 positive cases (0.8%; 95% CI: 0.4%-1.2%). However, assessment conducted on 3246 children in 2013 revealed five positives, giving point prevalence of 0.15%; 95% CI: 0.02%-0.28%. Four of the five children subjected to O-150 PCR proved negative. The data show that transmission of onchocerciasis has been interrupted based on national and WHO Guidelines of 2012 and 2016, respectively.

The Itwara onchocerciasis focus is located around the Itwara forest reserve in western Uganda. In 1991, annual treatments with ivermectin started in the focus. They were supplemented in 1995 by the control of the vector Simulium neavei, which was subsequently eliminated from the focus. The impact of the two interventions on the disease was assessed in 2010 by nodule palpations, examinations of skin snips by microscopy and PCR, and Ov16 recombinant ELISA. There was no evidence of any microfilaria in 688 skin snips and only 2 (0.06%) of 3316 children examined for IgG4 were slightly above the arbitrary cut off of 40. A follow up of the same children 21 months later in 2012 confirmed that both were negative for diagnostic antigen Ov-16, skin snip microscopy and PCR. Based on the World Health Organization (WHO) elimination criteria of 2001 and the Uganda onchocerciasis certification guidelines, it was concluded that the disease has disappeared from the Itwara focus after 19 years of ivermectin treatments and the elimination of the vector around 2001. Ivermectin treatments were recommended to be halted.

We review and analyze approaches over a 65 year period that have proven successful for onchocerciasis control in several different epidemiological settings. These include vector control with the goal of transmission interruption versus the use of mass drug administration using ivermectin (Mectizan®) monotherapy. Ivermectin has proven exceedingly effective because it is highly efficacious against Onchocerca volvulus microfilariae, the etiological agent of onchocercal skin and ocular disease and the infective stage for the vector. For these reasons, the drug was donated by the Merck Company for regional control programs in Africa and the Americas. Recurrent treatment with ivermectin at semi-annual intervals also impacts adult worms and result in loss of fecundity and increased mortality. Using a strategy of 6-monthly treatments with high coverage rates, the Onchocerciasis Elimination Program for the Americas has interrupted transmission in seven of the thirteen foci in the Americas and is on track to eliminate onchocerciasis in the region by 2015. Treatments given annually or semi-annually for 15–17 years in three hyperendemic onchocerciasis foci in Mali and Senegal also have resulted in a few infections in the human population with transmission levels below thresholds postulated for elimination. Follow-up evaluations did not detect any recrudescence of infection or transmission, suggesting that onchocerciasis elimination could be feasible with Mectizan® treatment in some endemic foci in Africa.

Brief Summary and Statement of Need: Pavement rehabilitation on Egan Drive from Mendenhall Loop Road to Brotherhood Bridge. Pavement rehabilitation on Glacier Highway from Brotherhood Bridge to the Auke Bay Ferry Terminal. The segment on Glacier Highway from Hamilton Drive to Fritz Cove Road was resurfaced in 2008 and is not included. This project contributes to the Department's Mission by reducing injuries, fatalities and property damage and by improving the mobility of people and goods. Funding: FY2009 FY2010 FY2011 FY2012 FY2013 FY2014 Total

We have used a two methods to attempt to isolate single-chain antibodies (scFv) specific for the surface of the Temecula strain of Xylella fastidiosa. A method using whole X. fastidosa cells to pan a phage display scFv library was successful and we recovered one very specific scFv that strongly interacts with Temecula and less so with the Tulare strain of X. fastidiosa. A method using a purified surface protein (mopB) from Temecula yielded two scFvs that interacted with that protein, but failed to interact with intact cells. Intended uses for these scFvs in a paratransgenesis method to control Pierce’s disease will be discussed. INTRODUCTION The glassy-winged sharpshooter (GWSS) is the principle vector of the xylem-limited bacterium Xylella fastidiosa (Xf), which causes Pierce’s disease (PD) in grapes. Limiting the spread of this pathogen by rendering GWSS incapable of pathogen transmission (paratransgenesis) is a promising method of pathogen control. Paratransgenesis seeks to modify the phenotype of an organism indirectly, by modifying its symbiotic bacteria to confer vector-incompetence Paratransgenic approaches to disrupt pathogen infection of humans are being developed by several groups. These include interference with the ability of triatomid bugs to transmit pathogens causing Chagas’ disease (Beard, Dotson et al. 2001), interference with HIV attachment to its target cells in the reproductive tracts of humans (Chang, Chang et al. 2003), and the elimination of persistent Candida infections from biofilms in chronically infected patients (Beninati, Oggioni et al. 2000). Paratransgenesis has also been applied to deliver cytokines mammalian guts to relieve colitis (Steidler, Hans et al. 2000; Steidler 2001). Thus, the method has wide applicability. Alcaligenes xylosoxidans denitrificans (Axd) is bacterial species that can colonize the GWSS foregut and cibarium, as well as various plant tissues, including xylem. It is non-pathogenic in insects and plants and is not known to be a pathogen in healthy humans. Given these characteristics, Axd has become the focus of our paratransgenesis efforts to control PD in grapes. Over the past two years we developed the technology to stably modify Axd by inserting genes into its chromosome. We have recently turned our attention to isolating factors that can specifically inhibit the transmission of Xf from GWSS. We report here the isolation of a single chain antibody phage that is specific for the surface of Xf and appears to have some strain specificity. Other tests have shown that this antibody phage can inhibit the transmission of PD by sharpshooters under experimental conditions. OBJECTIVES 1. Isolate single chain antibody (scFv) clones capable of binding to the surface of Xylella fastidiosa with high specificity either though the use of intact cells or purified surface proteins. 2. Test the strain specificity of these scFvs. RESULTS A. Single-chain antibodies (scFvs) as specific control agents of Xylella fastidiosa We have spent the past year designing and implementing a strategy to isolate factors that are specifically targeted at Xf for use in a paratransgenesis approach to controlling PD. The greatest degree of specificity against PD is likely to be obtained using synthetic antibody constructs called single-chain antibodies, or scFvs. An scFv is composed of a single gene consisting of the variable regions of an antibody heavy chain and light chain fused together with a synthetic linker sequence. Genes for these scFvs are randomized at certain codons and are constructed in large libraries that can be “panned” with an antigen of interest to obtain specific binders.

Xylella fastidiosa (Xf), which causes Pierce’s disease (PD) in grapevines, is transmitted by the glassy-winged sharpshooter (GWSS). Symbiotic control employs symbiotic bacteria to deliver anti-pathogen compounds to disrupt transmission of the pathogen to new host plants. Alcaligenes xylosoxidans denitrificans (Axd) was identified as a potential agent for paratransgenesis because it inhabits the foregut of GWSS and the xylem of plants, as does Xf. In this report, we describe the relationship between Axd (the symbiont), Xf (the plant pathogen), GWSS (the insect vector), and host plants to develop a delivery strategy for symbiotic control. Additionally, disruption of Xf-transmission by GWSS was demonstrated using two reagents, a single chained antibody fragment and an antibiotic peptide. INTRODUCTION The glassy-winged sharpshooter (GWSS) is the principal vector of the xylem-limited bacterium Xylella fastidiosa (Xf), which causes Pierce’s disease (PD) in grapevines. Limiting the spread of this pathogen by rendering GWSS incapable of pathogen transmission would control the disease. Symbiotic control approaches have been developed to disrupt Triatomid transmission of Trypanosoma cruzi [3], to prevent colitis in mammals [4, 11], and to interfere with transmission of HIV [8]. Candidate microbes that live in close proximity to the pathogen in the vector insects and in host plant tissues would by ideal vehicles to control Xf. Alcaligenes xylosoxidans denitrificans (Axd), originally isolated from the cibarium of GWSS, has been described as a nonpathogenic plant endophyte and a non-pathogenic soil-borne microbe [10, 12]. Axd, genetically marked with DsRed or EGFP protein, colonized the cibarium of GWSS for up to 35 days, the longest period tested [6]. Axd readily colonized the xylem vessels of several plants with citrus being the most hospitable to the bacterium. [5] Two categories of anti-pathogen reagents, single-chained antibodies (scFV) and antibiotic peptides, were tested for activity against Xf. Screening of scFV uncovered an antibody fragment that was specific to Xf and may be specific to the PD-causing strain of Xf. Four toxic peptides were identified that inhibited the growth of Xf, but did not inhibit the growth of Axd. OBJECTIVES 1. Identify relationships between Axd (the symbiont), Xf (the plant pathogen), GWSS (the insect vector), and host plants to develop a delivery strategy for symbiotic control. 2. Test the ability of anti-pathogens to disrupt Xf disease cycle. RESULTS Axd Movement and Colonization within Host Plants In two trials, chrysanthemums (n=20) were needle inoculated with DsRed Axd, which contains a kanamycin-resistance gene. One week later phloem and xylem fluid samples were collected independent of one another using a Scholander pressure bomb [7]. The collection resulted in about 20-50 μl of phloem fluid and 100-150 μl of xylem fluid per stem. DNA was extracted from the remaining half of each phloem and xylem fluid sample from each plant using the Extract-N-Amp Plant kit (Sigma Aldrich, Steinheim, Germany). Presence of Axd was then determined using QRT PCR. The other half of each phloem and xylem fluid sample was inoculated into LB broth containing kanamycin and incubated for 48 h at 37oC. After the incubation period, bacteria were screened for red fluorescence using a MZ12 fluorescent microscope (Leica Microsystems Inc., Heerbrugg, Switzerland). Positive samples were confirmed by QRT PCR. A higher proportion of xylem fluid samples tested positive for the presence of Axd than phloem samples in both trials: in trial 1 xylem 8/20, phloem 2/20 (א=4.8, 1df, p=0.0284); in trial 2 xylem 15/20, phloem 8/20 (א=5.013, 1df, p=0.025). In all

Group purchasing has become a widely used method for vari-ous and sundry groups to save money in many areas. This presen-tation will describe the development of an electric users’ group, itsuniqueness from other types of groups, savings members of thegroup have achieved, problems encountered along the way, oppor-tunities seized and prospects for the future.

One of the two onchocerciasis-control programmes in Africa is due to close by the end of 2002, so thought must be given to how the disease will be controlled in that continent. This disease is endemic in 37 countries (30 in Africa, six in the Americas, and Yemen), where it infects at least 17·7 million people and causes visual impairment in 500 000, and blindness (river blindness) in another 270 000. 1 WHO, Onchocerciasis and its controlReport of a WHO Expert Committee on Onchocerciasis Control. WHO Tech Rep Ser. 1995; 852: 1-104 PubMed Google Scholar , 2 Burnham G Onchocerciasis. Lancet. 1998; 351: 1341-1346 Summary Full Text Full Text PDF PubMed Scopus (101) Google Scholar Onchocerciasis is transmitted by black flies that breed in rapidly flowing rivers and streams. The adult forms of the parasite (Onchocerca volvulus) are long-lived (8–15 years), 3 Duke BOL The population dynamics of Onchocerca volvulus in the human host. Trap Med Parasitol. 1993; 44: 61-68 PubMed Google Scholar but it is the pre-larval stages (called microfilariae), released by female worms, that cause most of the disease.

The use of community residents as agents for distributing mass ivermectin therapy for onchocerciasis provides a component of community participation absent from mobile team delivery methods. Community-based distribution, however, presupposes preexisting human resources in the endemic villages capable of fulfilling the essential functions of an ivermectin distribution process: mobilizing and educating the population, dispensing the drug, maintaining records, and monitoring and treating adverse reactions. Even when such human resources exist, the community workers must continue to receive tangible support from both external (government and donor agencies) and internal (community) sources. Donor and government agencies must accept that their data collection demands will be limited by the literacy standards of the communities being served. Community leaders must agree to set and use their own local standards of payment (including food stuffs or exchange in kind) to compensate the distributors for their time and efforts. The use of locally available human and remunerative resources is a prerequisite for true community ownership of a program.

If ivermectin distribution programs are to have maximum impact on the morbidity and transmission of human onchocerciasis there must be broad and sustained acceptance within the endemic communities. Educational activities, developed with careful consideration of community attitudes, should promote positive treatment seeking behavior while simultaneously addressing local reservations about the control effort. To better understand the ambient knowledge, attitudes, and practices concerning onchocerciasis in the context of ivermectin use in Guatemala, we conducted a survey among 145 heads of households in five endemic communities. Given the country's long-standing nodulectomy program, it was not surprising that 100% of persons interviewed had heard of the disease 'la filaria', which they defined as a skin nodule that could cause blindness. Ninety-five percent of respondents identified surgery as the only cure for the condition. Relatively few (39%) knew that la filaria was caused by a worm, although slightly more (50%) knew that the condition was acquired by the bite of an insect. The term microfilaria was not broadly recognized. We also determined that onchocerciasis was not perceived as a serious health problem: few persons (12%) mentioned la filaria when requested to provide a complete list of illnesses that occurred in the community, and the gravity of infection (based on rank ordering of common illnesses) was similar to that of a bad cold. Recommendations were made which might assist long-term acceptance of a national chemotherapy initiative against onchocerciasis in Guatemala.

The International Drinking Water Supply and Sanitation Decade (1981-1990) has stimulated a movement to eradicate human infection with the helminthic parasite Dracunculus medinensis (dracunculiasis), whose victims are disabled for weeks or months during the painful emergence of one or more worms from beneath the skin. Each year, millions of people acquire this infection by drinking unclean water. Among the critical activities that are necessary for the elimination of dracunculiasis, one of the most fundamental is that of epidemiological surveillance. Surveillance activities play a key role in the strategy to target affected villages for improved water supplies and other control activities. Accurate surveillance data also stimulate interest and support for national eradication programmes. Dracunculiasis is a condition with excellent characteristics for reporting through passive surveillance systems. However, active surveillance, as well as other innovative surveillance strategies, should be used to establish baseline information in those villages where cases occur, and later to monitor epidemiologically important indices needed to evaluate the progress of elimination efforts.

The effect of averaging over various spatial scales (0.5–2.5° latitude) and times (1–24 h) on the relationship between the mean fraction of the averaging area covered by clouds colder than various IR equivalent blackbody temperature thresholds and the precipitation over that area is examined. While a linear relationship between fractional coverage and rainfall amount shows considerable scatter at the smallest scale, there is much better correspondence at the larger scales, with linear correlation coefficients often exceeding 0.8. Large-scale rainfall estimates based on linear regression coefficients detect the timing and magnitudes of major rainfall events during GATE. For scales on the order of 2–3° of latitude, estimates based on a linear model are comparable to those found by Stout et al. (1979) and Griffith et al. (1980) for the GATE area. This simple model appears to be limited to scales considerably larger than the convective scale. Averaging over these scales minimizes the effects of the spatial and temporal details of the convective fields. The linear model can be interpreted as the application of an effective mean rainfall rate to the entire precipitating cloudy area. While such an approach does not provide detailed resolution of the field of precipitation, estimation procedures based on linear models may be useful for large-scale budget studies and certain hydrologic and agricultural applications.

Journal of the American Society for Naval EngineersVolume 10, Issue 1 p. 193-200 COMPRESSED AIR EXPLOSIONS. Frank Richards, Frank RichardsSearch for more papers by this author Frank Richards, Frank RichardsSearch for more papers by this author First published: February 1898 https://doi.org/10.1111/j.1559-3584.1898.tb05773.xAboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onFacebookTwitterLinked InRedditWechat Volume10, Issue1February 1898Pages 193-200 RelatedInformation

Pierce's disease (PD) is caused by a xylem limited gram-negative Xylella fastidiosa (Xf) bacterium. Various species of sharpshooters, including the important glassy-winged sharpshooter (GWSS), transmit Xf. Currently, there is no cure for PD. Paratransgenesis is a new tool for the management of PD. Acquisition efficiency of GWSS to acquire Xf is about 80% when tested with Real-Time PCR. Results of selected phage antibody specific to Xf PD-strain to disrupt the pathogen are underway.