Animal Models,Prophylaxis, and Therapeutics for Arenavirus Infections

Arenaviruses are enveloped, bipartite negative single-stranded RNA viruses that can cause a wide spectrum of disease in humans and experimental animals including hemorrhagic fever. The majority of these viruses are rodent-borne and the arenavirus family can be divided into two groups: the Lassa-Lymphocytic choriomeningitis serocomplex and the Tacaribe serocomplex. Arenavirus-induced disease may include characteristic symptoms ranging from fever, malaise, body aches, petechiae, dehydration, hemorrhage, organ failure, shock, and in severe cases death. Currently, there are few prophylactic and therapeutic treatments available for arenavirus-induced symptoms. Supportive care and ribavirin remain the predominant strategies for treating most of the arenavirus-induced diseases. Therefore, efficacy testing of novel therapeutic and prophylactic strategies in relevant animal models is necessary. Because of the potential for person-to-person spread, the ability to cause lethal or debilitating disease in humans, limited treatment options, and potential as a bio-weapon, the development of prophylactics and therapeutics is essential. This article reviews the current arenavirus animal models and prophylactic and therapeutic strategies under development to treat arenavirus infection.     

Helminth antigens (Taenia solium, Taenia crassiceps, Toxocara canis, Schistosoma mansoni and Echinococcus granulosus) and cross-reactivities in human infections and immunized animals

Helminth antigens were investigated in the search for accessible heterologous antigens capable to discriminate different helminthiases, by the enzyme linked immunosorbent assay (ELISA) and the immunoblot assay (IB). Antigens used were: Taenia solium cysticercus total saline (Tso); Taenia crassiceps cysticercus vesicular fluid (Tcra-VF); T. crassiceps cysticercus glycoproteins (Tcra-GP and Tcra-(18-14)-GP); Toxocara canis larva excretory-secretory (TES); Schistosoma mansoni adult total saline (Sm) and Echinococcus granulosus hydatid fluid (Eg). The assayed sera were from patients with: cysticercosis (n = 18); toxocariasis (n = 40); schistosomiasis (n = 19) and hydatidosis (n = 50) with proven clinical and laboratory diagnosis, and sera from rabbits immunized with Tso, Tcra-VF, TES and Eg. Cross-reactivity occurred mostly between infections caused by Taenia and Echinococcus or in immunized rabbits, by ELISA. Moreover, the cross-reactivity among helminthiases was found with the use of antigens belonging to phylogenetically related parasite species, Eg, Tso and Tcra-VF, by sharing same antigenic components. Lower cross-reactivities were obtained by IB technique, when only peptides were considered as antigens, and the use of T. crassiceps purified glycoproteins demonstrated high sensitivity and specificity in the diagnosis of human cysticercosis, similarly to that using homologous antigen (Tso) by the same technique.     

Nosocomial fungal infections: epidemiology, infection control, and prevention

Fungal infections are an increasing cause of morbidity and mortality in hospitalized patients. This article reviews the current epidemiology of nosocomial fungal infections in adult patients, with an emphasis on invasive candidiasis and aspergillosis. Recently published recommendations and guidelines for the control and prevention of these nosocomial fungal infections are summarized.     

Hepatitis B,Hepatitis C and Human Immunodeficiency Virus Seropositivity Among Children in Kabul,Afghanistan: A Cross-Sectional Study

Background:

Hepatitis B virus (HBV), hepatitis C Virus (HCV), and human immunodeficiency virus (HIV) infections are significant causes of morbidity and mortality all over the world, especially in underdeveloped countries like Afghanistan. Limited data are available concerning the seroprevalence of HBV, HCV and HIV in the pediatric age group in Afghanistan .

Objectives:

The aim of the study was to assess HBV, HCV and HIV serology among children at an outpatient clinic in Kabul.

Patients and Methods:

A total number of 330 children were included to the study from outpatient clinics of Ataturk Kabul ISAF Role II Military Hospital from May to November 2012. Hepatitis B surface antigen (HBsAg), hepatitis C antibody (anti-HCV), and human immunodeficiency virus antibody (anti-HIV) were measured.

Results:

The mean age of children was 6.5 ± 4.2 years. The frequency of positive results for HBsAg, anti-HBs and anti-HCV in all age groups were 12 (3.6%), 47 (14.2%) and 2 (0.6%), respectively. Anti-HIV was not detected in any of the children''s serum samples. The frequency of positive results for HBsAg was significantly higher in children older than six years than in other age groups.

Conclusions:

Vaccination program including HBV has begun during the last five years in Afghanistan. The continuation of the vaccination program is of great importance. Vaccination program and implementation steps should be revised and the deficiencies, if any, should be overcome without delay.     

Laboratory and field evaluation of Teknar HP-D, a biolarvicidal formulation of Bacillus thuringiensis ssp. israelensis, against mosquito vectors

Larvicidal efficacy of Teknar HP-D, an improved biolarvicidal formulation of Bacillus thuringiensis ssp. israelensis (Bti), against Anopheles stephensi, Culex quinquefasciatus and Aedes aegypti was determined in the laboratory, and in field the efficacy of the formulation was tested against Cx. quinquefasciatus breeding in cesspits, unused wells and drains. The toxicity of the formulation to Gambusia affinis (larvivorous fish), Notonecta sp. and Diplonychus indicus (water bugs) was also evaluated in the laboratory. Teknar HP-D was field tested at three recommended dosages, 1, 1.5 and 2l/ha, selecting five habitats for each dosage. Another five habitats were kept untreated as controls. Ae. aegypti showed greatest susceptibility to the Bti toxin in the laboratory. In cesspits, all the three dosages caused >80% reduction of pupal recruitment up to day 6 post-treatment, indicating that a weekly application at the lowest would be necessary for sustained control. The residual activity of the formulation was longer in unused wells, causing >80% reduction of pupal recruitment for 17 days from the day of treatment. In controlling pupal recruitment the three dosages produced equal effect. Application of Teknar HP-D at 1 l/ha once in three weeks is therefore recommended to control Cx. quinquefasciatus in unused wells. However, in drains, >80% reduction of pupal recruitment was observed for only 3 days and hence, application of Teknar HP-D at 2 l/ha that caused significantly higher level of reduction twice in a week at 3-day interval is necessary. At dosages from 0.032 to 3.2 mg/l (ppm), Teknar HP-D was non-toxic to Gambusia fish. The two predatory water bugs, Notonecta sp. and Diplonychus indicus that fed on the surviving larvae of Cx. quinquefasciatus exposed to the sub-lethal doses (LC(50) and LC(80)) of Teknar HP-D were safe with out having any mortality.     

Laboratory and field efficacy of Pedalium murex and predatory copepod,Mesocyclops longisetus on rural malaria vector,Anopheles culicifacies

Objective

To test the potentiality of the leaf extract of Pedalium murex (P. murex) and predatory copepod Mesocyclops longisetus (M. longisetus) in individual and combination in controlling the rural malarial vector, Anopheles culicifacies (An. culicifacies) in laboratory and field studies.

Methods

P. murex leaves were collected from in and around Erode, Tamilnadu, India. The active compounds were extracted with 300 mL of methanol for 8 h in a Soxhlet apparatus. Laboratory studies on larvicidal and pupicidal effects of methanolic extract of P. murex tested against the rural malarial vector, An. culicifacies were significant.

Results

Evaluated lethal concentrations (LC₅₀) of P. murex extract were 2.68, 3.60, 4.50, 6.44 and 7.60 mg/L for I, II, III, IV and pupae of An. culicifacies, respectively. Predatory copepod, M. longisetus was examined for their predatory efficacy against the malarial vector, An. culicifacies. M. longisetus showed effective predation on the early instar (47% and 36% on I and II instar) when compared with the later ones (3% and 1% on III and IV instar). Predatory efficacy of M. longisetus was increased (70% and 45% on I and II instar) when the application was along with the P. murex extract.

Conclusions

Predator survival test showed that the methanolic extract of P. murex is non-toxic to the predatory copepod, M. longisetus. Experiments were also conducted to evaluate the efficacy of methanolic extract of P. murex and M. longisetus in the direct breeding sites (paddy fields) of An. culicifacies. Reduction in larval density was very high and sustained for a long time in combined treatment of P. murex and M. longisetus.     

Phages Preying on Bacillus anthracis,Bacillus cereus,and Bacillus thuringiensis: Past,Present and Future

Many bacteriophages (phages) have been widely studied due to their major role in virulence evolution of bacterial pathogens. However, less attention has been paid to phages preying on bacteria from the Bacillus cereus group and their contribution to the bacterial genetic pool has been disregarded. Therefore, this review brings together the main information for the B. cereus group phages, from their discovery to their modern biotechnological applications. A special focus is given to phages infecting Bacillus anthracis, B. cereus and Bacillus thuringiensis. These phages belong to the Myoviridae, Siphoviridae, Podoviridae and Tectiviridae families. For the sake of clarity, several phage categories have been made according to significant characteristics such as lifestyles and lysogenic states. The main categories comprise the transducing phages, phages with a chromosomal or plasmidial prophage state, γ-like phages and jumbo-phages. The current genomic characterization of some of these phages is also addressed throughout this work and some promising applications are discussed here.     

Longitudinal characterization of Acinetobacter baumannii-calcoaceticus complex, Klebsiella pneumoniae, and methicillin-resistant Staphylococcus aureus colonizing and infecting combat casualties

Drug-resistant Acinetobacter baumannii-calcoaceticus complex, Klebsiella pneumoniae, and methicillin-resistant Staphylococcus aureus colonize and infect combat casualties from Iraq and Afghanistan. We retrospectively evaluated relatedness, by pulsed-field gel electrophoresis and antibiotic susceptibility testing, of isolates colonizing and infecting casualties over 2 years. Colonizing organisms were unrelated to isolates producing later infection in up to 27% of cases; most isolates underwent change in antibiotic susceptibilities. The same is true for serial infecting isolates recovered during hospitalization.     

CYP1A1 , GSTM1 , GSTT1 and NQO1 polymorphisms and colorectal adenomas in Japanese men

AIM: To investigate the role of functional genetic poly-morphisms of metabolic enzymes of tobacco carcinogens in the development of colorectal adenomas. METHODS: The study subjects were 455 patients with colorectal adenomas and 1052 controls with no polyps who underwent total colonoscopy in a preretirement health examination at two Self Defense Forces hospitals. The genetic polymorphisms studied wereCYP1A1*2A (rs 4646903), CYP1A1*2C (rs 1048943), GSTM1 (null or non-null genotype), GSTT1 (null or non-null genotype) and NQO1 C609T (rs 1800566). Genotypes were determined by the polymerase chain reaction (PCR)-restriction fragment length polymorphism or PCR method using genomic DNA extracted from the buffy coat. Cigarette smoking and other life-style factors were ascertained by a self-administered questionnaire. The associations of the polymorphisms with colorectal adenomas were examined by means of OR and 95%CI, which were derived from logistic regression analysis. Statistical adjustment was made for smoking, alcohol use, body mass index and other factors. The gene-gene interaction and effect modification of smoking were evaluated by the likelihood ratio test. RESULTS: None of the five polymorphisms showed a significant association with colorectal adenomas, nor was the combination of GSTM1 and GSTT1 . A borderline significant interaction was observed for the combination of CYP1A1*2C and NQO1 (P = 0.051). The OR associated with CYP1A1*2C was significantly lower than unity among individuals with the NQO1 609CC genotype. The adjusted OR for the combination of the CYP1A1*2C allele and NQO1 609CC genotype was 0.61 (95%CI: 0.42-0.91). Although the interaction was not statistically significant (P = 0.24), the OR for individuals carrying the CYP1A1*2C allele and GSTT1 null genotype decreased significantly compared with those who had neither CYP1A1*2C allele nor GSTT1 null genotype (adjusted OR: 0.69, 95%CI: 0.49-0.97). Smoking did not modify the associations of the individual polymorphisms with colorectal adenomas. There w     

Antimicrobial activity of bismuth subsalicylate on Clostridium difficile,Escherichia coli O157:H7, norovirus,and other common enteric pathogens

Previous studies have shown bismuth subsalicylate (BSS) has antimicrobial properties, but few studies have addressed the mechanism of action. Furthermore, following BSS ingestion other bismuth salts form throughout the gastrointestinal tract including bismuth oxychloride (BiOCl) that also act upon enteric pathogens. To further understand the antimicrobial activity of bismuth in infectious diarrhea, the antimicrobial effect of BSS and BiOCl on Clostridium difficile, Salmonella, Shigella, Shiga toxin-producing Escherichia coli strains and norovirus (NoV) were measured. Bacterial enteric pathogens in pure culture or in human fecal material were exposed to 35mg/ml BSS or BiOCl with or without a vehicle suspension. BSS and BiOCl treated samples were quantified and visualized by transmission electron microscopy. To measure the effect on NoV, reduction of infectious murine NoV (MNV), a surrogate for human NoV, and Norwalk virus RNA levels were measured by viral plaque assay and RT-qPCR, respectively. BSS and BiOCl reduced bacterial growth by 3–9 logs in all strains with majority resulting in populations of <10 cfu/ml within 24 h. Similar results were found when fecal material was included. Microscopy images detected bismuth on bacterial membranes and within the bacterial organisms at 30 min post-treatment. At 8.8mg/ml BSS and BiOCl reduced infectivity of MNV significantly by 2.7 and 2.0 log after 24 h of exposure. In addition, both BSS and BiOCl slightly reduced the level of Norwalk replicon-bearing cells suggesting that bismuth may inhibit NoV in vivo. Collectively, our results confirm and build on existing data that BSS has antimicrobial properties against a wide-range of diarrhea-causing pathogens.     

Relation between clinical presentation, Helicobacter pylori density, interleukin 1beta and 8 production, and cagA status

BACKGROUND: It is not known whether cagA+ Helicobacter pylori in duodenal ulcer (DU) have enhanced virulence compared with non-DU cagA+ H pylori. AIMS: To investigate the relation between presentation, H pylori density, interleukin 1beta (IL-1beta) and IL-8 production, and cagA status. METHODS: Fifty DU and 50 gastritis patients with cagA+ H pylori and 11 with cagA- infections were studied. Bacterial density and cytokine production were assessed using the same biopsies. Cytokine production was also measured from supernatants of medium following coculture of H pylori with MKN-45 cells. RESULTS: There was no relation between H pylori density and cagA status. There was a dose dependent relation between mucosal cytokine levels and density of cagA+ H pylori. H pylori density increased to a threshold, followed by a rapid increase in cytokines and then a plateau. IL-1beta and IL-8 levels in the antrum were greater in DU than in gastritis; in the corpus the cytokine level/H pylori differed irrespective of similar H pylori densities. However, cytokine production was similar in vitro, independent of presentation or biopsy site, suggesting that host factors are critical determinants of the inflammatory response. Mucosal IL-8 and IL-1beta levels were low with cagA- and cagA+, cagE- H pylori infections. CONCLUSIONS: The increase in antral IL-1beta and IL-8 production and inflammation in DU is related to increased numbers of bacteria and not to an increase in cytokine production per cagA+ isolate. There was no evidence of enhanced virulence of H pylori from DU compared with cagA+ non-DU H pylori.     

Colchicine nonresponsiveness in familial Mediterranean fever: clinical, genetic, pharmacokinetic, and socioeconomic characterization

OBJECTIVES: To identify the ethnic, clinical, genetic, and pharmacokinetic correlates of colchicine treatment failure in patients with familial Mediterranean fever (FMF). METHODS: Fifty-nine FMF patients, unresponsive to a daily dose of > or =2 mg colchicine, were compared with 51 colchicine-responsive patients by clinical, demographic, and socioeconomic assessment, FMF gene (MEditerranean FeVer [MEFV]) mutation and serum amyloid A1 (SAA1) gene polymorphism analysis, and plasma and white blood cell colchicine level determination. RESULTS: Colchicine responders and nonresponders were comparable with respect to gender, age, duration and onset of the disease, and various demographic parameters. The 2 cohorts were found to carry mainly the M694V MEFV mutation and had a similar number of homozygotes or compound heterozygotes. Predominance of the alpha/beta alleles of SAA1 and comparable plasma and polymorphonuclear colchicine concentrations characterized both groups. Nonresponders were from lower socioeconomic backgrounds, had less education, and a more severe form of disease. A statistically significant 2-fold elevation of colchicine concentration in the mononuclear cells (MNC) of responders was found. CONCLUSIONS: Colchicine treatment failure in FMF is associated with inadequate colchicine MNC concentration, probably resulting from a genetic defect unrelated to the underlying FMF.     

Tribendimidine: a promising, safe and broad-spectrum anthelmintic agent from China

We review, for the first time, a 20-year Chinese story of research and development pertaining to tribendimidine, a promising anthelmintic agent that is safe and exhibits a broad spectrum of activity. Tribendimidine was first synthesized at the National Institute of Parasitic Diseases in Shanghai in the mid 1980s. In laboratory studies, tribendimidine showed high efficacy against Nippostrongylus braziliensis in rats, Necator americanus in hamsters, Ancylostoma caninum and Toxocara canis in dogs, and Syphacia mesocriceti in mice. Activity was also found against several species of cestodes in chicken. In clinical trials, a single oral dose of 400 mg tribendimidine, administered to patients infected only with N. americanus, or with N. americanus and Ancylostoma duodenalis, resulted in cure rates of 85.7% (132/154) and 89.8% (53/59), respectively. In comparison, a single oral dose of 400 mg albendazole resulted in significantly lower cure rates, namely 65.5% (91/139; chi(2) = 16.47, P < 0.001) and 71.7% (43/60; chi(2) = 6.29, P = 0.012), respectively. Single oral doses of tribendimidine (300 mg) and albendazole (400mg) were equally effective against Ascaris lumbricoides infections; cure rates were 96.0% (97/101) and 98.1% (101/103), respectively. In 5-14-year-old children with an Enterobius vermicularis infection, treated with a single oral dose of 200 mg tribendimidine, a cure rate of 81.6% (93/114) was observed. Tribendimidine was well-tolerated as only mild and transient side effects were observed. It would be of great public health significance if these findings are confirmed in other epidemiological settings, as more than one-quarter of the world population is currently affected by intestinal nematodes, with only very few drugs currently available on the market.     

Helminth infections and risk factor analysis among residents in Eryuan county, Yunnan province, China

Whilst infections with soil-transmitted helminths are common across China, the public-health significance of Schistosoma japonicum and food-borne helminths is more focalized. Only few studies have investigated the local epidemiology of helminth infections in rural China, including risk factor analysis. We collected stool and blood samples from 3220 individuals, aged 5-88 years, from 35 randomly selected villages in Eryuan county, Yunnan province, China. Stool samples were subjected to the Kato-Katz technique and examined for helminth eggs. Blood samples were tested for Trichinella spp., S. japonicum and cysticerci-specific antibodies. Data on individual and family-level risk factors were collected using questionnaires. The prevalence of Ascaris lumbricoides, Taenia spp., Trichuris trichiura and hookworms was 15.4%, 3.5%, 1.7% and 0.3%, respectively. The seroprevalence of Trichinella spp. was 58.8% and that of cysticercosis 18.5%. The egg positivity rate of S. japonicum in the 13 known endemic villages was 2.7%, and the corresponding seroprevalence was 49.5%. We observed a strong spatial heterogeneity in the families' economic status. S. japonicum infections were more prevalent among the Han than Bai nationality (odds ratio (OR)=3.77, 95% confidence interval (CI)=1.97-7.23) and tobacco growers (OR=3.66, 95% CI=1.77-7.60) and was only found at elevations below 2150 m above sea level. A. lumbricoides and Taenia spp. infections were more prevalent at altitudes above 2150 m when compared to lower settings (OR=1.51, 95% CI=1.24-1.84 and OR=5.32, 95% CI=3.42-8.28, respectively). The opposite was found for T. trichiura (OR=0.31, 95% CI=0.14-0.70). Our findings can guide the design and spatial targeting of control interventions against helminth infections in Eryuan county.     

Linking fecal bacteria in rivers to landscape,geochemical, and hydrologic factors and sources at the basin scale

Linking fecal indicator bacteria concentrations in large mixed-use watersheds back to diffuse human sources, such as septic systems, has met limited success. In this study, 64 rivers that drain 84% of Michigan’s Lower Peninsula were sampled under baseflow conditions for Escherichia coli, Bacteroides thetaiotaomicron (a human source-tracking marker), landscape characteristics, and geochemical and hydrologic variables. E. coli and B. thetaiotaomicron were routinely detected in sampled rivers and an E. coli reference level was defined (1.4 log₁₀ most probable number⋅100 mL⁻¹). Using classification and regression tree analysis and demographic estimates of wastewater treatments per watershed, septic systems seem to be the primary driver of fecal bacteria levels. In particular, watersheds with more than 1,621 septic systems exhibited significantly higher concentrations of B. thetaiotaomicron. This information is vital for evaluating water quality and health implications, determining the impacts of septic systems on watersheds, and improving management decisions for locating, constructing, and maintaining on-site wastewater treatment systems.Water quality degradation influenced by diffuse sources at large watershed scales has been difficult to describe. Human modifications of natural landscapes can permanently alter hydrologic cycles and affect water quality (1, 2). Deforestation (3) and increased impervious surface area (4) have been linked with decreased infiltration and thus increased surface runoff. Overland flows concentrate pollutants and rapidly transport them down gradient where they eventually enter surface water systems and affect water quality (5, 6). A number of models have been developed to calculate overland and surface water flows (7, 8) and nutrient/chemical transport (9), but few studies have focused on microbial movement from land to water, particularly nontraditional fecal indicator bacteria that can be used to track human sources of pollution.Microbial contamination poses one of the greatest health risks to swimming areas, drinking water intakes, and fishing/shellfish harvesting zones where human exposures are highest (10–12). These highly visible areas often receive more attention than sources of contamination because identifying the origin of pollution in complex watersheds requires costly comprehensive investigation of environmental and hydrologic conditions across temporal and spatial scales (13). Grayson et al. (14) suggest using a “snapshot” approach that captures water quality characteristics at a single point in time across broad areas to provide information frequently missed during routine monitoring. Compared with long-term comprehensive investigations, the snapshot approach reduces the number of samples, cost, and personnel required to examine pollution sources.Escherichia coli concentrations are commonly used to describe the relative human health risk during water quality monitoring in lieu of pathogen detection. Studies attempting to trace pollution in water back to a specific land use with E. coli have rarely produced definitive conclusions (15, 16). Using molecular approaches, specific source targets can be isolated in complex systems and have recently been used to investigate land use and water quality impairments (17). Furtula et al. (18) demonstrated ruminant, pig, and dog fecal contamination in an agriculturally dominated watershed (Canada) using Bacteroides markers. The Bacteroides thetaiotaomicron α-1–6 mannanase (B. theta) gene has a high human specificity (19–22), but no studies to date have linked its presence to land use patterns.Reference conditions have been established for minimally disturbed environments based on measurements of macroinvertebrates, fish, and diatoms (23–25), but microbial reference conditions have not been adequately explored or defined. Based on 15 unimpaired California streams, microbial reference conditions for E. coli [1.0 log₁₀ most probably number (MPN)⋅100 mL⁻¹] and enterococci (1.2 log₁₀ MPN⋅100 mL⁻¹) were defined as being below state water quality thresholds (26). In the Great Lakes, a human health threshold of 2.37 log₁₀ E. coli MPN⋅100 mL⁻¹ (27), or a level equally protective of human health, has been adopted by all state governments. However, this health-associated reference level was derived from epidemiological studies undertaken at beaches throughout the United States (28, 29) with limited knowledge of local implications.In response to water quality degradation from human stressors and the poorly understood microbial conditions in large-scale fresh water systems such as the Great Lakes basin, this paper aims to (i) examine the spatial distribution of E. coli and a human specific source marker (B. theta) in 64 river systems that drain most of the state’s Lower Peninsula under baseflow conditions, (ii) identify baseflow reference levels of fecal contamination in rivers, and (iii) determine how key chemical, physical, environmental, hydrologic, and land use variables are linked to river water quality at large scales.